国药学中科大

1 国药 学中 科大 备 课 笔 记 本 院部： 药学院 教研室： 药物分析 课程名称： 药物分析实验与指导 教学对象：药物分析方向、药理方向和临床药学方向 教师姓名： 杭太俊 中国药科大学教务处制 2 Lecture notes Department： Pharmacy Staff room： Pharmaceutical analysis Course name： Pharmaceutical Analysis Experiments Teaching object： Pharmaceutical analysis Pharmacology and clinical pharmacy Teacher name： Tai-jun Hang 3 1、葡萄糖的性状、鉴别和检查 教学目的 1. 了解药品鉴别、检查的目的和意义； 2. 掌握药品性状测定方法和性状的正确描述； 3. 掌握药品的常用鉴别的方法和原理； 4. 掌握药品中一般杂质检查的方法原理和限量计算方法。 本章讲授内容 【性状】 本品为无色结晶或白色结晶性或颗粒性粉末；无臭，味甜。 本品在水中易溶，在乙醇中微溶。 比旋度测定 取本品约 10g，精密称定，置 100ml量瓶中，加水适量与氨试液 0.2ml， 溶解后，用水稀释至刻度，摇匀，放置 10分钟，在 25℃时，依法测定（中国药典 2000年 版二部附录Ⅵ E），比旋度为＋52.5°至＋53.0°。 【鉴别】 (1)取本品约 0.2g，加水 5ml溶解后，缓缓滴入温热的碱性酒石酸铜试液中，即 生成氧化亚铜的红色沉淀。 【检查】 酸度 取本品 2.0g，加水 20ml 溶解后，加酚酞指示液 3 滴与氢氧化钠滴定液 (0.02mol/L)0.20ml，应显粉红色。 氯化物 取本品 0.6g，依法检查（中国药典 2000年版二部附录Ⅷ A），与 标准 excel标准偏差excel标准偏差函数exl标准差函数国标检验抽样标准表免费下载红头文件格式标准下载 氯化钠 溶液 6.0ml制成的对照液比较，不得更浓(0.01％)。 重金属 取本品 4.0g，加水 23ml溶解后，加醋酸盐缓冲液(pH3.5)2ml，依法检查（中 国药典 2000年版二部附录Ⅷ H第一法），含重金属不得过百万分之五。 砷盐 取本品 2.0g，加水 5ml 溶解后，加稀硫酸 5ml 与溴化钾溴试液 0.5ml，置水浴上加 热约 20分钟，使保持稍过量的溴存在，必要时，再补加溴化钾溴试液适量，并随时补充蒸 散的水分，放冷，加盐酸 5ml 与水适量使成 28ml，依法检查（中国药典 2000 年版二部附 录Ⅷ J第一法），应符合规定(0.0001％)。 砷盐 取本品 2.0g，加水 5ml 溶解后，加稀硫酸 5ml 与溴化钾溴试液 0.5ml，置水浴 上加热约 20分钟，使保持稍过量的溴存在，必要时，再补加溴化钾溴试液适量，并随时补 充蒸散的水分，放冷，加盐酸 5ml 与水适量使成 28ml，依法检查（中国药典 2000 年版二 部附录Ⅷ J第一法），应符合规定(0.0001％)。 4 Experiment 1 The Description， Identification and Tests of Glucose 1. Purpose 1. To understand the aims and purposes of the drug quality control. 2. To learn about the methods for the determination and the special words for the description of the characteristics of drugs. 3. To experiment on the identification of glucose. 4. To study and experiment on the methods of drug tests 2. Contents and teach time assignment Description Colourless crystals or a white crystalline or granular powder； odourless； taste， sweet. Freely soluble in water； slightly soluble in ethanol. Specific optical rotation Dissolve， about 10 g， weighed accurately， with a quantity of water and 0.2 ml of ammonia TS in a 100 ml volumetric flask and dilute with water to volume. Mix well and allow to stand for 10 minutes. The specific optical rotation of the resulting solution is +52.5o ~ +53.0o at 25℃(Appendix VI E)#. Identification (1) Dissolve about 0.2 g in 5 ml of water， add dropwise hot alkaline cupric tartrate TS； a red precipitate of cuprous oxide is produced. Acidity Dissolve 2.0 g in 20 ml of water, add 3 drops of phenolphthalein IS and 0.20 ml of sodium hydroxide (0.02 mol/L) VS; a pink colour is produced. Chloride Carry out the limit test for chlorides (Appendix VIII A)， using 0.60 g. Any opalescence produced is not more pronounced than that of a reference using 6.0 ml of sodium chloride standard solution (0.01%). Heavy metals Dissolve 4.0 g in 23 ml of water， add 2 ml of sodium acetate BS (pH 3.5)， carry out the limit test for heavy metals (Appendix VIII H， method 1): not more than 0.0005%. # : Appendix refers to the Volumn 2 of Chinese Pharmacopoeia unless otherwise stated. 5 Arsenic Dissolve 2.0 g in 5 ml of water， add 5 ml of dilute sulfuric acid and 0.5 ml of potassium bromide-bromine TS. Heat on a water bath for about 20 minutes and maintain the presence of excess of bromine. Add a quantity of potassium bromide-bromine TS， if necessary. Replace the evaporated water constantly and cool， then add 5 ml of hydrochloric acid and dilute with water to 28 ml. The solution complies with the limit test for arsenic (Appendix VIII J， method 1) (0.0001%). 6 2、阿司匹林及阿司匹林肠溶片的质量分析 教学目的 1. 掌握质量检验的项目与方法； 2. 掌握阿司匹林及阿司匹林肠溶片分析的操作条件及要点。 本章讲授内容 【性状】本品为白色结晶或结晶性粉末；无臭或微带醋酸臭，味微酸；遇湿气即缓缓水 解。 【鉴别】 (1)取本品约 0.1g，加水 10ml，煮沸，放冷，加三氯化铁试液 1滴，即显紫堇色。 (2)取本品约 0.5g，加碳酸钠试液 10ml，煮沸 2分钟后，放冷，加过量的稀硫酸，即析 出白色沉淀，并发生醋酸的臭气。 【检查】 溶液的澄清度 取本品 0.50g，加温热至约 45℃的碳酸钠试液 10ml溶解后，溶液应澄 清。 游离水杨酸 取本品 0.10g，加乙醇 1ml 溶解后，加冷水适量使成 50ml，立即加新制 的稀硫酸铁铵溶液〔取盐酸溶液(9→100)1ml，加硫酸铁铵指示液 2ml后，再加水适量使成 100ml〕1ml，摇匀；30 秒钟内如显色，与对照液(精密称取水杨酸 0.1g，加水溶解后，加 冰醋酸 1ml，摇匀，再加水使成 1000ml，摇匀，精密量取 1ml，加乙醇 1ml、水 48ml与上 述新制的稀硫酸铁铵溶液 1ml，摇匀)比较，不得更深(0.1％)。 【含量测定】 原料 取本品约 0.4g，精密称定，加中性乙醇(对酚酞指示液显中性)20ml 溶解后，加 酚酞指示液 3 滴，用氢氧化钠滴定液(0.1mol/L)滴定。每 1ml氢氧化钠滴定液(0.1mol/L)相 当于 18.02mg的 C9H8O4。 片剂 取本品 10 片，研细，用中性乙醇 70ml，分数次研磨，并移入 100ml 量瓶中， 充分振摇，再用水适量洗涤研钵数次，洗液合并于 100ml 量瓶中，再用水稀释至刻度，摇 匀，滤过，精密量取滤液 10ml(相当于阿司匹林 0.3g)，置锥形瓶中，加中性乙醇(对酚酞指 示液显中性)20ml，振摇，使阿司匹林溶解，加酚酞指示液3滴，滴加氢氧化钠滴定液(0.1mol/L) 至溶液显粉红色，再精密加氢氧化钠滴定液(0.1mol/L)40ml，置水浴上加热 15 分钟并时时 振摇，迅速放冷至室温，用硫酸滴定液(0.05mol/L)滴定，并将滴定的结果用空白试验校正。 每 1ml氢氧化钠滴定液(0.1mol/L)相当于 18.02mg C9H8O4。 7 Experiment 2 The Analysis of Aspirin and Aspirin Enteric-coated Tablets 1. Purpose 1. To learn about the procedures and the items for drug analysis. 2. To experiment on the analysis of Aspirin and its Enteric-coated Tablets. 2. Contents and teach time assignment Description White crystals or a white crystalline powder； odourless or with a faint acetic acid odour； taste， faintly sour； gradually hydrolyses in contact with moisture to form salicylic acid and acetic acid； aqueous solution yields an acidic reaction Identification (1) To about 0.1 g add 10 ml of water， boil and cool. Add 1 drop of ferric chloride TS； a violet colour is produced. (2) To about 0.5 g add 10 ml of sodium carbonate TS， boil for 2 minutes and cool. Add dilute sulfuric acid in excess； a white precipitate is produced and an odour of acetic acid is perceptible. Clarity of solution A solution of 0.50 g in 10 ml of sodium carbonate TS preheated to about 45℃ is clear. Salicylic acid Dissolve 0.10 g in 1 ml of ethanol， add cold water to produce 50 ml， add immediately 1 ml of freshly prepared dilute ferric ammonium sulfate solution [ to 1 ml of hydrochloric acid solution (9 →100) add 2 ml of ferric ammonium sulfate IS and add water to produce 100 ml] and mix well； any colour produced in 30 seconds is not more intense than that of a reference preparation (dissolve 0.1 g of salicylic acid， accurately weighed， in water， add 1 ml of glacial acetic acid， mix well， add water to produce 1000 ml and mix well. Measure accurately 1 ml， add 1 ml of ethanol， 48 ml of water and 1 ml of freshly prepared dilute ferric ammomium sulfate solution and mix well) (0.1%). Assay Material Dissolve about 0.4 g， accurately weighed， in 20 ml of neutral ethanol (neutral to phenolphthalein IS) and add 3 drops of phenolphthalein IS. Titrate with sodium hydroxide (0.1 mol/L) VS. Each ml of sodium hydroxide (0.1 mol/ L) VS is equivalent to 18.02 mg of C9H8O4. Tablet Weigh accurately and powder 10 tablets， triturate with divided portions of 70 ml neutral ethanol and transfer to a 100 ml volumetric flask， shake thoroughly， wash the mortar several times with sufficient water， add the combine washings to the flask， dilute 8 with water to volume， mix well and filter. Measure accurately 10 ml of the filtrate equivalent to about 0.3 g of aspirin into a conical flask， add 20 ml of neutral ethanol (neutral to phenophthalein IS)， shake well. Add 3 drops of phenophthalein IS and sodium hydroxide (0.1 mol/L) VS dropwise until the solution becomes pink. Add 40 ml of sodium hydroxide (0.1 mol/L) VS， accurately measured， heat on a water bath for 1 minutes with shaking， cool immediately to room temperature and titrate with sulfuric acid(0.05 mol/L) VS. Perform a bank determination and make any necessary correction. Each ml of sodium hydroxide(0.1 mol/L) VS is equivalent to 18.02 mg of C9H8O4. 9 3、复方磺胺甲噁唑片的质量分析 教学目的 1. 熟悉复方制剂双波长计算分光光度方法含量测定原理； 2. 掌握复方磺胺甲噁唑片实验的操作条件及要点。 本章讲授内容 【性状】本品为白色片。标示量等特性――――（2min） 【鉴别】 (1)取本品的细粉适量（约相当于磺胺甲噁唑 50mg），显芳香第一胺的鉴别 反应（中国药典 2000年版二部附录Ⅲ）。 (2)取本品的细粉适量（约相当于甲氧苄啶 50mg），加稀硫酸 10ml，微热溶解后，放冷， 滤过，滤液加碘试液 0.5ml，即生成棕褐色沉淀。――――――化学反应式（15min） 【含量测定】 ―――――――――――（25min） 磺胺甲噁唑 取本品 10片，精密称定，研细，精密称取适量（约相当于磺胺甲噁唑 50mg 与甲氧苄啶 10mg），置 100ml 量瓶中，加乙醇适量，振摇 15 分钟使磺胺甲噁唑与甲氧苄 啶溶解，加乙醇稀释至刻度，摇匀，滤过，取续滤液作为供试品溶液；另精密称取在 105 ℃干燥至恒重的磺胺甲噁唑对照品 50mg与甲氧苄啶对照品 10mg，分别置 100ml量瓶中， 各加乙醇溶解并稀释至刻度，摇匀，分别作为对照品溶液(1)与对照品溶液(2)。精密量取供 试品溶液与对照品溶液(1)、(2)各 2ml，分别置 100ml 量瓶中，各加 0.4％氢氧化钠溶液稀 释至刻度，摇匀，照分光光度法（中国药典 2000年版二部附录Ⅳ A），取对照品溶液(2)的 稀释液，以 257nm 为测定波长（λ2），在 304nm 波长附近（每间隔 0.5nm）选择等吸收 点波长为参比波长（λ1），要求ΔA＝Aλ2-Aλ1＝0。再在λ2与λ1波长处分别测定供试 品溶液的稀释液与对照品溶液(1)的稀释液的吸收度，求出各自的吸收度差值（ΔA），计算， 即得。 甲氧苄啶 仪器狭缝不得大于 1nm。如使用自动扫描仪，波长重现性不得大于 0.2nm， 如使用手动仪器时，波长调节器应同一方向旋转并时时用对照液核对等吸收点波长。精密量 取上述供试品溶液与对照品溶液(1)、(2)各 5ml，分别置 100ml量瓶中，各加盐酸-氯化钾溶 液（取 0.1mol/L盐酸溶液 75ml与氯化钾 6.9g，加水至 1000ml，摇匀）稀释至刻度，摇匀， 照分光光度法（中国药典 2000年版二部附录Ⅳ A），取对照品溶液(1)的稀释液，以 239.0nm 为测定波长（λ2），在 295nm波长附近（每间隔 0.2nm）选择等吸收点波长为参比波长（λ 1），要求ΔA＝Aλ2－Aλ1＝0，再在λ2 与λ1 波长处分别测定供试品溶液的稀释液与对 照品溶液(2)的稀释液的吸收度，求出各自的吸收度差值（ΔA），计算，即得。 10 Experiment 3 The Analysis of Aspirin and Aspirin Enteric-coated Tablets 1. Purpose 1. To study the spectro-photometric method for the simultaneous determination of drug components in compound formulation. 2. To experiment on the assay of Compound Sulfamethoxazole Tablets by the spectro-photometric method. 2. Contents and teach time assignment Description White tablets Identification (1) Yields the reactions characteristic of primary aromatic amines (Appendix III )， using a quantity of the powdered tablets equivalent to about 50 mg of Sulfamethoxazole. (2) To a quantity of the powdered tablets equivalent to about 50 mg of trimethoprim add 10 ml of dilute sulfuric acid， heat gently to dissolve， cool and filter. To the filtrate add 0.5 ml of iodine TS， a dark brown precipitate is produced. Assay Sulfamethoxazole Weigh accurately and powder 10 tablets. Weigh accurately a quantity of the powder equivalent to about 50 mg of sulfamethoxazol and 10 mg of trimethoprim into a 100 ml volumetric flask， add a quantity of ethanol and shake for 15 minutes to dissolve sulfamethoxazole and trimethoprim， dilute with ethanol to volume and shake well. Filter and use the successive filtrate as the test solution. Transfer 50 mg of sulfamethoxazole CRS and 10 mg of trimethoprim CRS， previously dried to constant weight at 105℃ and accurately weighed， separately to two 100 ml volumetric flasks， dissolve in ethanol and dilute to volume， mix well. The resulting solutions are used as reference solution (1) and (2) respectively. Transfer 2 ml each of the test solution and reference solutions， accurately measured， to three 100 ml volumetric flasks separately， dilute with 0.4% sodium hydroxide solution to volume and mix well. Measure the absorbance of diluted reference solution (2) at 257 nm (λ2) (AppendixIV A) and find out an isobestic point (λ1) near 304 nm with a stepwise difference of 0.5 nm， so that 12 λλ AA − =0. Measure the absorbance of diluted test solution and that of diluted reference solution (1) at wavelengths λ2 and λ1， calculate the difference in absorbance (∆A ) for each solution and the content of C10H11N3O3S. 11 Trimethoprim Transfer 5 ml each of the test solution and reference solutions， accurately measured， to three 100 ml volumetric flasks separately， dilute with hydrochloric acid-potassium chloride solution [To 75 ml of hydrochloric acid (0.1 mol/L) VS and 6.9 g of potassium chloride add water to produce 1000 ml， mix well] to volume and mix well. Measure the absorbance of diluted reference solution (1) at 239 nm (λ2) (Appendix IV A) and find out an isobestic point (λ1) near 295 nm with a stepwise difference of 0.2 nm， so that 12 λλ AA − =0. Measure the absorbance of diluted test solution and that of diluted reference solution (2) at wavelengths λ2 and λ1， calculate the difference in absorbance (∆A) for each solution and the content of C14H18N4O3. 12 4、血浆中阿司匹林的高效液相色谱法测定 教学目的 1. 了解生物样品测定的过程； 2. 掌握阿司匹林血浆样品测定的方法和步骤。 本章讲授内容 1. 标准溶液 阿司匹林标准溶液(100µg/ml)： 取阿司匹林对照品约 10mg，精密称定，置 100ml量瓶中，以乙腈溶解，并稀释至刻度， 摇匀，在 4℃保存备用。 邻甲基苯甲酸(内标)标准溶液(50µg/ml)： 取邻甲基苯甲酸对照品约 5mg，精密称定，置 100ml量瓶中，以乙腈溶解，并稀释至 刻度，摇匀，在 4℃保存备用。 2. 血浆样品处理 用肝素化的试管收集健康志愿受试者静脉血液，立即离心分取血浆，于-20℃以下低温 保存，待测。 阿司匹林血浆样品处理 取冷冻的血浆样品在冰水浴中解冻，精密吸取 0.5ml置 1.5ml 的离心管中，精密加入邻甲基苯甲酸标准溶液 10µl，加 0.5mol/L盐酸 0.1ml和乙腈 0.5ml， 涡旋 1分钟，4℃放置 15分钟后，4℃下 12000 rpm离心 10分钟，分取上清液 0.5ml，加 氯化钠 0.1g，涡旋 5秒，4℃静置 10分钟后，12000 rpm离心 10分钟，分取上清液作为 供试液。 3. 标准曲线 精密吸取市售空白血浆 0.5ml，分别置 1.5ml 的离心管中，精密加入阿司匹林标准溶 液适量，使阿司匹林的浓度分别为 0、0.2、0.4、0.8、2.0、4.0、8.0和 20.0 µg/ml。然后 照“血浆样品处理”项下方法，自“精密加入邻甲基苯甲酸内标溶液 10µl”起，同法处理， 并照“阿司匹林血药浓度测定法”测定。将阿司匹林与内标邻甲基苯甲酸的峰面积比，对血 浆中阿司匹林的浓度进行线性回归，即得。 4. 阿司匹林血药浓度测定回收率和精密度试验 O CH3 O OH O OH CH3 O 13 精密吸取市售空白血浆 0.5ml，分别置 1.5ml 的离心管中，精密加入阿司匹林标准溶 液适量，制成阿司匹林的浓度分别为 0.2、2.0和 20.0 µg/ml的低、中、高浓度的血浆样品 各 5 份。然后照“血浆样品处理”项下方法，自“加 0.5mol/L 盐酸 0.1ml”起，至“分取 上清液 0.5ml”止，同法处理；再分别精密加入邻甲基苯甲酸标准溶液 10µl，加氯化钠 0.1g， 涡旋 5秒，4℃静置 10分钟后，12000 rpm离心 10分钟，分取上清液作为低、中、高浓度 的供试液。 另取水 0.5ml，分别置 1.5ml 的离心管中，精密加入阿司匹林标准溶液适量，制成阿 司匹林的浓度分别为 0.2、2.0和 20.0 µg/ml的低、中、高浓度的对照样品各 2份。分别照 上述步骤同样操作，分取上清液作为低、中、高浓度的对照液。 取供试液和对照液，照“阿司匹林血药浓度测定”项下的条件分别测定，按内标法，依 不同浓度水平，分别以峰面积比进行计算，即得阿司匹林血浆样品测定低、中、高浓度的回 收率和精密度，应符合规定。 5. 阿司匹林血药浓度测定 照高效液相色谱法(中国药典 2000年版二部附录Ⅴ D)测定。 色谱条件与系统适用性试验 用十八烷基硅烷键合硅胶为填充剂；以 1%磷酸-乙腈 (70∶30)为流动相；流速为每分钟 1ml；检测波长为 237nm。取取空白样品和“标准曲线” 项下的阿司匹林 20.0µg/ml样品分别测定，阿司匹林与邻甲基苯甲酸的分离度应符合规定， 理论板数按阿司匹林峰计算应不低于 3000；空白样品色谱中，在阿司匹林与邻甲基苯甲酸 位置应没有干扰。 测定法 取阿司匹林血浆样品供试液 20µl，注入液相色谱仪， 记录 混凝土 养护记录下载土方回填监理旁站记录免费下载集备记录下载集备记录下载集备记录下载 色谱图，按内标法， 以标准曲线进行计算即得。 14 Experiment 4 Determination of aspirin(ASA) in human plasma by HPLC 1. Purpose 1. To learn about the principles and procedures for the determination of drugs in biological samples. 2. To experiment on the determination of aspirin in human plasma. 2. Contents 1. Standard solutions Aspirin standard solution(ASA，100μg/ml): Weigh about 10 mg of aspirin RS， accurately， to a 100 ml volumetric flask， dissolve and dilute with acetonitrile to volume， mix. Keep at 4℃ for future use. 2-methyl benzoic acid (internal standard)(MBA，50μg/ml): Weigh about 5mg of 2-methyl benzoic acid RS， accurately， to a 100 ml volumetric flask， dissolve and dilute with acetonitrile to volume， and mix. Keep at 4℃ for further use. 2. Sample preparation Blood samples were collected freshly in a heparinized centrifuge tube from healthy volunteers and the plasma samples were separated immediately by centrifugation and stored at -20℃ until analysis. Aspirin plasma sample preparation Thaw the frozen plasma sample in an ice-water bath. Transfer accurately 0.5 ml of it to a 1.5 ml microcentrifuge tube and add accurately 10μl of MBA standard solution. Then add 0.1 ml of 0.5 mol/L hydrochloric acid and 0.5 ml of acetonitrile. Vortex the mixture for 1 minute and keep at 4℃ for 15 minutes. Then centrifuge at 12000rpm for 10 min at 4℃. To 0.5 ml of the supernatant add 0.1g of sodium chloride and vortex for 5s and placed at 4℃for 10min before centrifuge at 12， 000rpm for 10 min at 4℃. Use the supernatant for the determination of aspirin. 3. Calibration curve To each of 0.5 ml of blank plasma in 1.5ml microcentrifuge tubes， add a suitable quantity of ASA standard solution to make the concentrations of ASA to be 0， 0.2， 0.4， 0.8， 2.0， 4.0， 8.0， and 20.0μg/ml， respectively. Then process them in the same O CH3 O OH O OH CH3 O 15 way as directed under the ‘sample preparation’ beginning with ‘add accurately 10μl of MBA standard solution’. Then make the determination according to the method of the ‘determination of ASA in human plasma’. Establish the calibration curve by plotting the peak-area ratios of ASA to internal standard versus its concentrations. 4. Recovery and precision for ASA determination To each of the 1.5 ml microcentrifuge tubes， add accurately 0.5ml of blank plasma and a suitable quantity of ASA standard solution to make the ASA concentration in plasma to be 0.2， 2.0， or 20.0μg/ml， respectively(n=5). Treat the samples according to the procedures under the ‘sample preparation’ beginning with ‘Then add 0.1 ml of 0.5 mol/L hydrochloric acid’ until ‘To 0.5 ml of the supernatant add 0.1g of sodium chloride’. Then add 10μl of MBA standard solution to each of the solutions and vortex for 5s， place the tubes at 4℃ for 10min before centrifuge at 12，000rpm for 10 min. Use the supernatant as test solution. At the same time， transfer 0.5 ml of water into each of 1.5 ml microcentrifuge tubes， add accurately a suitable amount of ASA standard solution to make the ASA concentrations to be 0.2， 2.0， or 20.0μg/ml， respectively(n=2). Treat them in the same way as just described. Use the supernatants as reference solutions. Then make the determination according to the procedures described under the ‘determination of ASA in human plasma’. The mean recovery and precision of different concentrations， calculated by comparison of the peak-area ratios of ASA to MBA in the test solutions with those of standards at the same concentration， must comply with the requirements. 5. Determination of Aspirin in human plasma System suitability Carry out the method for high performance liquid chromatography (Appendix V D) using a column packed with octadecylsilane bonded silica gel and a mixture of 1% phosphoric acid-acetonitrile (70:30) as the mobile phase. The flow rate is about 1 ml per minute and detection wavelength is 220 nm. Chromatograph the blank sample and the calibration curve solution which contains 20μ g/ml ASA， separately， and record the peak responses. The resolution between ASA and MBA complies with the related requirements， and the number of theoretical plates of the column is not less than 3000， calculated with reference to the peak of ASA. There is no interference in the blank sample chromatogram at the position of ASA and MBA. Determination Inject 20μl of ASA plasma sample into the column， and record the chromatogram. Calculate the ASA concentration in the plasma with respect to the peak area ratios obtained in the chromatogram by the internal standard method according to the calibration curve. 16 5、维生素 E胶丸的含量测定 教学目的 1. 掌握气相色谱法测定的原理与方法； 2. 掌握维生素 E含量的操作条件及要点。 本章讲授内容 维 生 素 E (Vitamin E，C31H52O3，FW=472.75) 本品为(±)-2，5，7，8-四甲基-2-(4，8，12-三甲基十三烷基)-6-苯并二氢吡喃醇醋酸 酯。含 C31H52O3应为 96.0%～102.0％。 【性状】 本品为微黄色或黄色透明的黏稠液体；几乎无臭；遇光色渐变深。 本品在无水乙醇、丙酮、乙醚或石油醚中易溶，在水中不溶。 【含量测定】 取装量差异项下的内容物，混合均匀，精密称取适量（约相当于维生素 E 20mg），置棕色具塞锥形瓶中，照维生素 E项下的方法，精密加入内标溶液 10ml，密塞， 摇匀，取 1～3μl注入气相色谱仪，并依法测定校正因子，计算，即得。 照气相色谱法(中国药典 2000年版二部附录Ⅴ E)测定。 色谱条件与系统适用性试验 以硅酮(OV-17)为固定相，涂布浓度为 2％；柱温为 265 ℃。理论板数按维生素 E峰计算应不低于 500，维生素 E峰与内标物质峰的分离度应大于 2。 校正因子测定 取正三十二烷适量，加正己烷溶解并稀释成每 1ml中含 1.0mg的溶液， 摇匀，作为内标溶液。另取维生素 E对照品约 20mg，精密称定，置棕色具塞锥形瓶中，精 密加入内标溶液 10ml，密塞，振摇使溶解，取 1～3μl注入气相色谱仪，计算校正因子。 O OH CH3 CH3 CH3 CH3 CH3 CH3CH3 CH3 17 Experiment 5 The Assay of Vitamin E 1. Purpose 1. To study the principles and procedures of gas chromatography for the assay of volatile drugs. 2. To experiment on the assay of Vitamin E. 2. Contents Vitamin E (Alpha Tocopheryl Acetate) (C31H52O3， FW=472.75， [59-02-9] ) Vitamin E is dl-2，5，7，8-tetramethyl-2-(4，8，12-trimethyl tridecyl)chromanyl-6-acetate. It contains not less than 96.0% and not more than 102.0% of C31H52O3. Description A clear pale yellow or yellow viscous， oily liquid； almost odourless； the colur deepens on exposure to light. Freely soluble in dehydrated ethanol， acetone， ether or petroleum ether； insoluble in water. Assay Mix well the contents obtained in the test for Weight variation. To a quantity， accurately weighed， of the mixed contents equivalent to 20 mg of vitamin E add 10 ml， accurately measured， of internal standard solution in an amber coloured flask stoppered， shake well. Carry out the Assay described under vitamin E， injecting 1~3µl of the solution into the column and determine