药分英文部分1

药分英文部分1 绪论 GLP (Good Laboratory Practice) 药品非临床研究质量管理规范 Good Laboratory Practice (GLP) deals with the organization, process and conditions under which laboratory studies are planned, performed, monitored, recorded and reported. GLP practices are intended to promote the quality and validity of test data. Published GLP regulations and guidelines have a significant impact on the daily operation of an analytical laboratory. GLP is a regulation. It is not only good analytical practice. Good analytical practice is important, but it is not enough. For example, the laboratory must have a specific organizational structure and procedures to perform and document laboratory work. The objective is not only quality of data but also traceability and integrity of data. But the biggest difference between GLP and Non-GLP work is the type and amount of documentation. For a GLP inspector it should be possible to look at the documentation and to easily find out who has done a study, how the experiment was carried out, which procedures have been used, and whether there has been any problem and if so how it has been solved. All routine work should follow written standard operating procedures (SOP). Facilities such as laboratories should be large enough and have the right construction to ensure the integrity of a study, for example, to avoid cross contamination Test and control articles should have the right quality and instruments should be calibrated and well maintained People should be trained or otherwise qualified for the job Raw data and other data should be acquired, processed and archived to ensure integrity of data. GMP(Good Manufacture Practice) 药品生产质量管理规 GMP refers to the Good Manufacturing Practice Regulations promulgated by the US Food and Drug Administration under the authority of the Federal Food, Drug, and Cosmetic Act These regulations, which have the force of law, require that manufacturers, processors, and packagers of drugs, medical devices take proactive steps to ensure that their products are safe, pure, and effective. GMP regulations require a quality approach to manufacturing, enabling companies to minimize or eliminate instances of contamination, mixups, and errors. This in turn, protects the consumer from purchasing a product which is not effective or even dangerous. Failure of firms to comply with GMP regulations can result in very serious consequences including recall, seizure, fines, and jail time. GMP regulations address issues including recordkeeping, personnel qualifications, sanitation, cleanliness, equipment verification, process validation, and complaint handling. Most GMP requirements are very general and open-ended, allowing each manufacturer to decide individually how to best implement the necessary controls. GCP (Good Clinical Practice) 药品临床试验管理规范 GCP Good Clinical Practice (GCP) is an international quality standard that is provided by International Conference on Harmonisation (ICH), an international body that defines standards, which governments can transpose into regulations for clinical trials involving human subjects. Good Clinical Practice guidelines include protection of human rights as a subject in clinical trial. It also provides assurance of the safety and efficacy of the newly developed compounds. Good Clinical Practice Guidelines include standards on how clinical trials should be conducted, define the roles and responsibilities of clinical trial sponsors, clinical research investigators, and monitors. In the pharmaceutical industry monitors are often called Clinical Research Associates. GAP(Good Agricultural Practice) 中药材生产质量管理规范 AQC 分析质量管理 第十一章 吩噻嗪类抗精神病药物的分析 Perphenazine Injection Perphenazine Injection is a sterile solution of Perphenazine in Water for Injection, prepared with the aid of Citric Acid. It contains not less than 90.0 percent and not more than 110.0 percent of the labeled amount of C21H26ClN3OS, as the citrate Packaging and storage— Preserve in single-dose or multiple-dose containers, preferably of Type I glass, protected from light. USP Reference standards— USP Endotoxin RS. USP Perphenazine RS. NOTE—Throughout the following procedures, protect test or assay specimens, the USP Reference Standard, and solutions containing them, by conducting the procedures without delay, under subdued light, or using low-actinic glassware. Identification— Dilute 1 mL with methanol to 5 mL. Apply 5 µL each of this solution and a solution of USP Perphenazine RS in methanol containing 1 mg per mL to a suitable thin-layer chromatographic plate, coated with a 0.25-mm layer of chromatographic silica gel. Develop the chromatogram in a solvent system consisting of a mixture of acetone and ammonium hydroxide (200:1) until the solvent front has moved about 15 cm. Air-dry the plate, and spray lightly with a solution of iodoplatinic acid prepared by dissolving 100 mg of chloroplatinic acid in 1 mL of 1 N hydrochloric acid, adding 25 mL of potassium iodide solution (4 in 100), diluting with water to 100 mL, and adding 0.50 mL of formic acid: the RF value of the principal spot obtained from the Injection corresponds to that obtained from the Standard solution. Bacterial endotoxins— It contains not more than 35.7 USP Endotoxin Units per mg of perphenazine. pH : between 4.2 and 5.6. Other requirements— It meets the requirements under Injections. Assay— Acid-alcohol solution— Transfer 10 mL of hydrochloric acid to a 1000-mL flask containing 500 mL of alcohol and 300 mL of water. Dilute with water to volume. Palladium chloride solution— Dissolve 100 mg of palladium chloride in a mixture of 1 mL of hydrochloric acid and 50 mL of water in a 100-mL volumetric flask, heating on a steam bath to effect solution. Cool, dilute with water to volume, and mix. Store in an amber bottle and use within 30 days. On the day of use, transfer 50 mL to a 500-mL volumetric flask, add 4 mL of hydrochloric acid and 4.1 g of anhydrous sodium acetate, dilute with water to volume, and mix. Standard preparation— Dissolve an accurately weighed quantity of USP Perphenazine RS in Acid-alcohol solution to obtain a solution having a known concentration of about 150 µg per mL. Assay preparation— Dilute 3.0 mL of Injection with Acid-alcohol solution to 100 mL in a volumetric flask. Procedure— Mix 10.0 mL each of the Assay preparation and the Standard preparation with 15.0 mL of Palladium chloride solution, filter, if necessary, and concomitantly determine the absorbances of these solutions, against a reagent blank, in 1-cm cells at the wavelength of maximum absorbance at about 480 nm, with a suitable spectrophotometer. Calculate the quantity, in mg, of C21H26ClN3OS in the volume of Injection taken by the formula: 0.1C(AU / AS), in which C is the concentration, in µg per mL, of USP Perphenazine RS in the Standard preparation, and AU and AS are the absorbances of the solutions from the Assay preparation and the Standard preparation, respectively 第九章 二氢吡啶类钙通道阻滞剂的分析-双语课程(Nifedipine ) C17H18N2O6 346.33 3,5-Pyridinedicarboxylic acid, 1,4-dihydro-2,6-dimethyl-4-(2-nitrophenyl)-, dimethyl ester. Dimethyl 1,4-dihydro-2,6-dimethyl-4-(o-nitrophenyl)-3,5-pyridinedicarboxylate [21829-25-4]. ? Nifedipine contains not less than 98.0 percent and not more than 102.0 percent of C17H18N2O6, calculated on the dried basis. Packaging and storage— Preserve in tight, light-resistant containers. USP Reference standards — USP Nifedipine RS. USP Nifedipine Nitrophenylpyridine Analog RS. USP Nifedipine Nitrosophenylpyridine Analog RS. NOTE—Nifedipine, when exposed to daylight and certain wavelengths of artificial light, readily converts to a nitrosophenylpyridine derivative. Exposure to UV light leads to the formation of a nitrophenylpyridine derivative. Perform assays and tests in the dark or under golden fluorescent or other low-actinic light. Use low-actinic glassware. A: Infrared Absorption —Do not dry specimens. B: Ultraviolet Absorption — Spectral range: 450 to 200 nm. Solution— To a 10-mL volumetric flask containing 14 mg of Nifedipine add 1.0 mL of chloroform, dilute with methanol to volume, and mix. Pipet a 1.0-mL aliquot of the solution into a 100-mL volumetric flask, dilute with methanol to volume, and mix. C: The retention time of the major peak in the chromatogram of the Assay preparation corresponds to that in the chromatogram of the Standard preparation, as obtained in the Assay. Melting range, : between 171 and 175. Loss on drying— Dry it at 105 to constant weight: it loses not more than 0.5% of its weight. Residue on ignition: not more than 0.1%, an ignition temperature of 600 being used. Heavy metals, Method II : 0.001%. Perchloric acid titration— Transfer about 4 g of Nifedipine, accurately weighed, to a 250-mL conical flask, and dissolve in 160 mL of glacial acetic acid with the aid of an ultrasonic bath. Add 3 drops of p-naphtholbenzein TS, and titrate to a green endpoint with 0.1 N perchloric acid VS: not more than 0.12 mL of 0.1 N perchloric acid is consumed for each g of Nifedipine. Chloride and Sulfate— To 5.0 g of Nifedipine in a 140-mL beaker add 4.0 mL of 6 N acetic acid and 46 mL of water. Bring carefully to a boil on a hot plate, cool, and filter through paper free of chloride and sulfate. Use this Nifedipine filtrate for the following tests. Chloride— Pipet 2.5 mL of the Nifedipine filtrate into a 50-mL color-comparison tube, and add 12.5 mL of water. Into a matched color-comparison tube pipet 10 mL of a Standard solution containing 8.2 µg of sodium chloride per mL corresponding to 5 µg of chloride per mL, add 5.0 mL of water, and mix. To each tube add 0.15 mL of 0.3 M nitric acid and 0.3 mL of silver nitrate TS, and mix: the opalescence exhibited by the Nifedipine filtrate does not exceed that of the Standard solution (0.02%). Sulfate— Pipet into each of two 50-mL matched color-comparison tubes 1.5 mL of sulfate solution consisting of sufficient potassium sulfate dissolved in water to obtain a sulfate concentration of 10 µg per mL. To each tube add, successively and with continuous shaking, 0.75 mL of alcohol, 0.5 mL of a 6.1% aqueous solution of barium chloride, and 0.25 mL of 6 N acetic acid. Shake for an additional 30 seconds. Pipet into one tube, designated the Standard tube, 15 mL of the sulfate solution. Pipet into the other tube, designated the Specimen tube, 3 mL of the Nifedipine filtrate and 12 mL of water: the turbidity exhibited by the Specimen tube does not exceed that of the Standard tube (0.05%). 第十一章 抗生素类药物的分析(Ampicillin ) C16H19N3O4S (anhydrous) 349.41 Ampicillin is anhydrous or contains three molecules of water of hydration. It contains not less than 900 µg and not more than 1050 µg of C16H19N3O4S per mg, calculated on the dried basis. Packaging and storage— Preserve in tight containers. Crystallinity : meets the requirements. Bacterial endotoxins— Where the label states that Ampicillin is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it contains not more than 0.15 USP Endotoxin Unit per mg of ampicillin. pH : between 3.5 and 6.0, in a solution containing 10 mg per mL. Water, Method I : not more than 2.0% where it is labeled as Ampicillin (anhydrous); between 12.0% and 15.0% where it is labeled as Ampicillin (trihydrate). Assay—Mobile phase— Prepare a suitable filtered and degassed mixture of water, acetonitrile, 1 M monobasic potassium phosphate, and 1 N acetic acid (909:80:10:1). Make adjustments if necessary (see System Suitability under Chromatography). Diluent— Mix 10 mL of 1 M monobasic potassium phosphate and 1 mL of 1 N acetic acid, dilute with water to 1000 mL, and mix. Standard preparation— Dissolve a suitable quantity of USP Ampicillin RS, accurately weighed, in Diluent to obtain a solution having a known concentration of about 1 mg per mL, using shaking and sonication, if necessary, to achieve complete dissolution. Use this solution promptly after preparation. Assay preparation— Transfer an accurately weighed quantity of Ampicillin, equivalent to about 100 mg of anhydrous ampicillin, to a 100-mL volumetric flask, add about 75 mL of Diluent, shake and sonicate, if necessary, to achieve complete dissolution, dilute with Diluent to volume, and mix. Use this solution promptly after preparation. Chromatographic system (see Chromatography)— The liquid chromatograph is equipped with a 254-nm detector, a 4-mm × 5-cm pre-column containing 5- to 10-µm packing L1, and a 4-mm × 30-cm analytical column containing 5- to 10-µm packing L1. The flow rate is about 2 mL per minut 第十二章 喹啉与青蒿素类抗疟药物的分析(Quinine Sulfate ) (C20H24N2O2)2?H2SO4?2H2O 782.94 Cinchonan-9-ol, 6?-methoxy-, (8a,9R)-, sulfate (2:1) (salt), dihydrate. Quinine sulfate (2:1) (salt) dihydrate [6119-70-6]. Quinine Sulfate is the sulfate of an alkaloid obtained from the bark of species of Cinchona. It contains not less than 99.0 percent and not more than 101.0 percent of total alkaloid salt, calculated as (C20H24N2O2)2?H2SO4, on the anhydrous basis. Packaging and storage— Preserve in well-closed, light-resistant containers. USP Reference standards — USP Quinine Sulfate RS. USP Quininone RS. Identification— A: A 1 in 2000 solution in dilute sulfuric acid (1 in 350) exhibits a vivid blue fluorescence. On the addition of a few drops of hydrochloric acid, the fluorescence disappears. B: In the test for Chromatographic purity, the RF value of the principal spot obtained from the Test preparation corresponds to that from the Standard preparation. C: A solution (1 in 50) made with the aid of a few drops of hydrochloric acid responds to the tests for Sulfate. Specific rotation: between -235 and-245 . Test solution: 20 mg per mL, in 0.1 N hydrochloric acid. Water, Method : between 4.0% and 5.5%. Residue on ignition: not more than 0.1%. Heavy metals, Method II : 0.001%. Chloroform-alcohol-insoluble substances— Warm 2 g with 15 mL of a mixture of chloroform and dehydrated alcohol (2:1) at about 50 for 10 minutes. Filter through a tared, sintered-glass filter, using gentle suction. Wash the filter with five 10-mL portions of the chloroform-alcohol mixture, dry at 105 for 1 hour, and weigh: the weight of the residue does not exceed 2 mg (0.1%). Chromatographic purity— Standard preparation— Prepare a solution of USP Quinine Sulfate RS in diluted alcohol to contain 6 mg per mL. Diluted standard preparation— Dilute a portion of the Standard preparation with diluted alcohol to a concentration of 0.06 mg per mL. Related substances preparation— Prepare a solution in diluted alcohol containing in each mL 0.05 mg each of USP Quininone RS (corresponding to 0.06 mg of the sulfate), and 0.10 mg of cinchonidine (corresponding to 0.12 mg of the sulfate). solution of Test preparation— Prepare a Quinine Sulfate in diluted alcohol to contain 6 mg per mL. Procedure— Apply 10-µL portions of the Test preparation, the Standard preparation, the Diluted standard preparation, and the Related substances preparation to a suitable thin-layer chromatographic plate (see Chromatography) coated with a 0.25-mm layer of chromatographic silica gel. Allow to dry, and develop the chromatogram in a solvent system consisting of a mixture of chloroform, acetone, and diethylamine (5:4:1), the solvent chamber being used without previous equilibration. When the solvent front has moved about 15 cm, remove the plate from the chamber, mark the solvent front, and allow the solvent to evaporate. Spray the chromatogram with glacial acetic acid. Locate the spots on the plate by examination under long-wavelength UV light. Any spot produced by the Test preparation at the RF value of a spot produced by the Related substances preparation is not greater in size or intensity than that corresponding spot. Apart from these spots and from the spot appearing at the RF value of Quinine Sulfate, any additional fluorescent spot is not greater in size or intensity than the spot of the Diluted standard preparation. Spray the plate with potassium iodoplatinate TS. Any spot produced by the Test preparation is not greater in size or intensity than a corresponding spot from the Related substances preparation. 第十三章 莨菪烷类抗胆碱药物的分析 (Atropine Sulfate Injection) ? Atropine Sulfate Injection is a sterile solution of Atropine Sulfate in Water for Injection. It contains not less than 93.0 percent and not more than 107.0 percent of the labeled amount of (C17H23NO3)2?H2SO4? H2O. Packaging and storage— Preserve in single-dose or in multiple-dose containers, preferably of Type I glass. USP Reference standards— USP Atropine Sulfate RS. USP Endotoxin RS. Identification (see Thin-Layer Chromatographic Identification Test)— Adsorbent: chromatographic silica gel. Developing solvent: mixture of chloroform and diethylamine (9:1). Test preparation— Use undiluted. Apply 15 µL. Detection reagent: potassium iodoplatinate TS. Procedure— Proceed as directed for Procedure under Thin-Layer Chromatographic Identification Test, the spots on the plate located by spraying with Detection reagent. Bacterial endotoxins— It contains not more than 55.6 USP Endotoxin Units per mg of atropine sulfate. pH: between 3.0 and 6.5. Other requirements— It meets the requirements under Injections. Assay— Acetate buffer— Prepare a solution in water containing in each L 0.05 mole of sodium acetate and 2.9 mL of glacial acetic acid. Mobile phase— Transfer 5.1 g of tetrabutylammonium hydrogen sulfate to a 1-L volumetric flask, add 50 mL of acetonitrile, and dilute with Acetate buffer to volume. Adjust with 5 N sodium hydroxide to a pH of 5.5 ? 0.1. Standard preparation— Dissolve an accurately weighed quantity of USP Atropine Sulfate RS in water, and dilute quantitatively with water to obtain a solution having a known concentration of about 80 µg per mL. Assay preparation— Transfer an accurately measured volume of Injection, equivalent to about 2 mg of atropine sulfate, to a 25-mL volumetric flask, dilute with water to volume, and mix. Resolution solution— Prepare a solution in water containing about 2.5 µg of p-hydroxybenzoic acid per mL. Dilute one volume of this solution with four volumes of the Standard preparation. Chromatographic system (see Chromatography)— The liquid chromatograph is equipped with a 254-nm detector and 30-cm × 3.9-mm column that contains packing L1. The flow rate is about 2 mL per minute. Chromatograph the Standard preparation, and record the peak responses as directed for Procedure: the relative standard deviation for replicate injections is not more than 1.5%. In a similar manner, chromatograph the Resolution solution: the retention time of p-hydroxybenzoic acid is about 1.6 relative to that of atropine, and the resolution, R, between the p-hydroxybenzoic acid and atropine peaks is not less than 2.2. Procedure— Separately inject equal volumes (about 100 µL) of the Standard preparation and the Assay preparation into the chromatograph, record the chromatograms, and measure the areas for the major peaks. Calculate the quantity, in mg, of (C17H23NO3)2?H2SO4?H2O in each mL of the Injection taken by the formula: (694.85/676.83)(25C/V)(rU / rS), in which 694.85 and 676.83 are the molecular weights of atropine sulfate monohydrate and anhydrous atropine sulfate, respectively; C is the concentration, in mg per mL, of USP Atropine Sulfate RS in the Standard preparation; V is the volume, in mL, of Injection taken; and rU and rS are the peak responses obtained from the Assay preparation and the Standard preparation, respectively. 第十四章 维生素类药物的分析(Vitamin A) Vitamin A contains a suitable form of retinol (C20H30O; vitamin A alcohol) and possesses vitamin A activity equivalent to not less than 95.0 percent of that declared on the label. It may consist of retinol or esters of retinol formed from edible fatty acids, principally acetic and palmitic acids. It may be diluted with edible oils; or it may be incorporated in solid, edible carriers or excipients; and it may contain suitable antimicrobial agents, dispersants, and antioxidants. Packaging and storage— Preserve in tight containers, preferably under an atmosphere of an inert gas, protected from light. Labeling— Label it to indicate the form in which the vitamin is present, and to indicate the presence of any antimicrobial agent, dispersant, antioxidant, or other added substance, and to indicate the vitamin A activity in terms of the equivalent amount of retinol, in mg per g. The vitamin A activity may be stated also in USP Units, on the basis that 1 USP Vitamin A Unit equals the biological activity of 0.3 µg of the all-trans isomer of retinol. USP Reference standards— USP Vitamin A RS. Identification— A: To 1 mL of a chloroform solution of it containing the equivalent of approximately 6 µg of retinol, add 10 mL of antimony trichloride TS: a transient blue color appears at once. B: Thin-Layer Chromatographic Identification Test Test solution— FOR LIQUID FORM OF VITAMIN A— Dissolve a volume equivalent to about 15,000 USP Units in chloroform to obtain 10 mL of solution. FOR SOLID FORM OF VITAMIN A— Weigh a quantity equivalent to about 15,000 USP Units, place in a separator, add 75 mL of water, shake vigorously for 1 minute, extract with 10 mL of chloroform by shaking for 1 minute, and centrifuge to clarify the chloroform extract. Standard solution— Dissolve the contents of 1 ampul of USP Vitamin A RS in chloroform to obtain 25.0 mL. Developing solvent system: a mixture of cyclohexane and ether (4:1). Procedure— Apply at the starting point of the chromatogram 0.015 mL of the Standard solution and 0.01 mL of the Test solution, and proceed as directed for Thin-Layer Chromatography under Chromatography. Allow the solvent front to move a distance of 10 cm, remove the plate, and air-dry. Spray with phosphomolybdic acid TS: the blue-green spot formed is indicative of the presence of retinol. The approximate RF values of the predominant spots, corresponding to the different forms of retinol, are 0.1 for the alcohol form, 0.45 for the acetate, and 0.7 for the palmitate. Absorbance ratio— The ratio of the corrected absorbance (A325) to the observed absorbance A325 determined as directed under Vitamin A Assay is not less than 0.85. Assay— Using a suitable quantity of Vitamin A, accurately weighed, proceed as directed under Vitamin A Assay. Aspirin Aspirin C9H8O4 180.16 Benzoic acid, 2-(acetyloxy)-. Salicylic acid acetate [50-78-2]. Aspirin contains not less than 99.5 percent and not more than 100.5 percent of C9H8O4, calculated on the dried basis. Packaging and storage— Preserve in tight containers. USP Reference standards — USP Aspirin RS. Identification— A: Heat it with water for several minutes, cool, and add 1 or 2 drops of ferric chloride TS: a violet-red color is produced. B: Infrared Absorption (197K). Loss on drying (731)— Dry it over silica gel for 5 hours: it loses not more than 0.5% of its weight. Mix and accurately weigh the substance to be tested, and, unless otherwise directed in the individual monograph, conduct the determination on 1 to 2 g. If the test specimen is in the form of large crystals, reduce the particle size to about 2 mm by quickly crushing. Tare a glass-stoppered, shallow weighing bottle that has been dried for 30 minutes under the same conditions to be employed in the determination. Put the test specimen in the bottle, replace the cover, and accurately weigh the bottle and the contents. By gentle, sidewise shaking, distribute the test specimen as evenly as practicable to a depth of about 5 mm generally, and not more than 10 mm in the case of bulky materials. Place the loaded bottle in the drying chamber, removing the stopper and leaving it also in the chamber. Dry the test specimen at the temperature and for the time specified in the monograph. Upon opening the chamber, close the bottle promptly, and allow it to come to room temperature in a desiccator before weighing. Readily carbonizable substances (271)— Dissolve 500 mg in 5 mL of sulfuric acid TS: the solution has no more color than Matching Fluid Q. Residue on ignition(281): not more than 0.05%. Substances insoluble in sodium carbonate TS— A solution of 500 mg in 10 mL of warm sodium carbonate TS is clear. Chloride 221— Boil 1.5 g with 75 mL of water for 5 minutes, cool, add sufficient water to restore the original volume, and filter. A 25-mL portion of the filtrate shows no more chloride than corresponds to 0.10 mL of 0.020 N hydrochloric acid (0.014%). Sulfate 211— Dissolve 6.0 g in 37 mL of acetone, and add 3 mL of water. Titrate potentiometrically with 0.02 M lead perchlorate(高氯酸铅), prepared by dissolving 9.20 g of lead perchlorate in water to make 1000 mL of solution. not more than 1.25 mL of 0.02 M lead perchlorate(高氯酸铅) is consumed (0.04%). Heavy metals— Dissolve 2 g in 25 mL of acetone, and add 1 mL of water. Add 1.2 mL of thioacetamide-glycerin硫代乙酰胺base TS and 2 mL of pH 3.5 Acetate Buffer, and allow to stand for 5 minutes: any color produced is not darker than that of a control made with 25 mL of acetone and 2 mL of Standard Lead Solution (see Heavy Metals), treated in the same manner. The limit is 10 µg per g. Limit of free salicylic acid— Dissolve 2.5 g in sufficient alcohol to make 25.0 mL. To each of two matched color-comparison tubes add 48 mL of water and 1 mL of a freshly prepared, diluted ferric ammonium sulfate solution硫酸铁铵(prepared by adding 1 mL of 1 N hydrochloric acid to 2 mL of ferric ammonium sulfate TS and diluting with water to 100 mL) Into one tube pipet 1 mL of a standard solution of salicylic acid in water, containing 0.10 mg of salicylic acid per mL. Into the second tube pipet 1 mL of the 1 in 10 solution of Aspirin. Mix the contents of each tube: after 30 seconds, the color in the second tube is not more intense than that in the tube containing the salicylic acid (0.1%). Assay— Place about 1.5 g of Aspirin, accurately weighed, in a flask, add 50.0 mL of 0.5 N sodium hydroxide VS, and boil the mixture gently for 10 minutes. Add phenolphthalein(酚酞) TS, and titrate the excess sodium hydroxide with 0.5 N sulfuric acid VS. Perform a blank determination (see Residual Titrations under Titrimetry \). Each mL of 0.5 N sodium hydroxide is equivalent to 45.04 mg of C9H8O4. Phenobarbital Phenobarbital C12H12N2O3 232.24 2,4,6(1H,3H,5H)-Pyrimidinetrione, 5-ethyl-5-phenyl-5-Ethyl-5-phenylbarbituric acid [50-06-6]. Phenobarbital contains not less than 98.0 percent and not more than 101.0 percent of C12H12N2O3, calculated on the dried basis. Packaging and storage— Preserve in well-closed containers. USP Reference standards (11)— USP Phenobarbital RS. Identification A: The IR absorption spectrum of a potassium bromide dispersion of it exhibits maxima only at the same wavelengths as that of a similar preparation of USP Phenobarbital RS. If a difference appears, dissolve portions of both the test specimen and the USP Reference Standard in a suitable solvent, evaporate the solutions to dryness, and repeat the test on the residues. B: The retention time of the major peak in the chromatogram of the Assay preparation corresponds to that of the Standard preparation, both relative to the internal standard, as obtained in the Assay. Melting range (741): between 174 and 178, but the range between beginning and end of melting does not exceed 2. Loss on drying (731)— Dry it at 105 for 2 hours: it loses not more than 1.0% of its weight. Residue on ignition (281): not more than 0.15%. Organic volatile impurities, Method V (467): meets the requirements. Assay— pH 4.5 Buffer solution— Dissolve about 6.6 g of sodium acetate trihydrate(三水醋酸钠) and 3.0 mL of glacial acetic acid冰醋酸 in 1000 mL of water, and adjust, if necessary, with glacial acetic acid to a pH of 4.5 ? 0.1. Mobile phase— Prepare a filtered and degassed mixture of pH 4.5 Buffer solution and methanol (3:2), making adjustments if necessary. Internal standard solution— Dissolve a sufficient quantity of caffeine in a mixture of methanol and pH 4.5 Buffer solution (1:1) to obtain a solution having a concentration of about 125 µg per mL. Standard preparation— Dissolve about 20 mg of USP Phenobarbital RS, accurately weighed, in 15.0 mL of Internal standard solution. Sonicate if necessary. Assay preparation— Transfer about 20 mg of Phenobarbital, accurately weighed, to a conical flask, add 15.0 mL of Internal standard solution, mix, and sonicate for 15 minutes. Filter through a membrane filter (0.5 µm or finer porosity) before use. Chromatographic system (see Chromatography (621))— The liquid chromatograph is equipped with a 254-nm detector and a 4-mm × 25-cm column that contains packing L1. The flow rate is about 2 mL per minute. Chromatograph the Standard preparation, and record the peak responses as directed for Procedure: the resolution, R, between the analyte and the internal standard peaks is not less than 1.2, the tailing factor for the analyte and the internal standard peaks is not greater than 2.0, and the relative standard deviation for replicate injections is not more than 2.0%. Procedure— Separately inject equal volumes (about 10 µL) of the Standard preparation and the Assay preparation into the chromatograph, record the chromatograms, and measure the responses for the major peaks. The relative retention times are about 0.6 for caffeine and 1.0 for phenobarbital. Calculate the quantity, in mg, of C12H12N2O3 in the portion of Phenobarbital taken by the formula: W(RU / RS) in which W is the weight, in mg, of USP Phenobarbital RS taken for the Standard preparation, and RU and RS are the peak response ratios obtained from the Assay preparation and the Standard preparation, respectively. Secobarbital C12H18N2O3 238.28 2,4,6(1H,3H,5H)-Pyrimidinetrione, 5-(1-methylbutyl)-5- (2-propenyl)-. 5-Allyl-5-(1-methylbutyl)barbituric acid [76-73-3]. Secobarbital contains not less than 97.5 percent and not more than 100.5 percent of C12H18N2O3, calculated on the dried basis. Packaging and storage— Preserve in tight containers. USP Reference standards— USP Secobarbital RS. Identification, Infrared Absorption. Loss on drying— Dry it over silica gel for 18 hours: it loses not more than 1.0% of its weight. Residue on ignition : not more than 0.1%. Organic volatile impurities, Method V : meets the requirements. Solvent— Use dimethyl sulfoxide. Isomer content— Dissolve about 300 ? 5 mg in 5 mL of sodium hydroxide solution (1 in 100), add a solution of 300 ? 5 mg of p-nitrobenzyl bromide in 10 mL of alcohol, reflux for 30 minutes, cool, collect the precipitate on a small filter, and wash with water: the precipitate, recrystallized from 25 mL of alcohol and dried at 105 for 30 minutes, melts between 156 and 161. Assay— Add about 450 mg of Secobarbital, accurately weighed, to 60 mL of dimethylformamide in a 125-mL conical flask. Add 4 drops of thymol(麝香草酚) blue TS, and titrate with 0.1 N sodium methoxide VS, using a magnetic stirrer and taking precautions against the absorption of atmospheric carbon dioxide. Perform a blank determination, and make any necessary correction. Each mL of 0.1 N sodium methoxide is equivalent to 23.83 mg of C12H18N2O3.