【作者】 朱凯；

【导师】 范琦； 张小松；

【作者基本信息】 重庆医科大学 ， 药物分析， 2007， 硕士

【摘要】 目的:双氢青蒿素哌喹片为青蒿素第二代抗疟疾药,是治疗疟疾,尤其是多重抗药性恶性疟疾的首选药物。其两个有效成分分别为:双氢青蒿素（Dihydroartemisinin,DHA）和磷酸哌喹（Piperaquine phosphate,PQP）。我国现行国家药品质量标准中,DHA的含量测定方法为:用硅胶柱分离洗脱后,挥干溶剂,收集残渣,经碱水解产生有紫外吸收的物质,紫外分光光度法测定。该操作步骤多,引入误差的机会多。本课题旨在建立专属性强、准确、简便的定量分析方法,采用HPLC-UV法直接测定双氢青蒿素哌喹片中的DHA。我国现行国家药品质量标准中,采用紫外分光光度法（Ultraviolet Specterphotometry,UV）测定双氢青蒿素哌喹片中PQP的含量。由于复方制剂中干扰因素多,不经分离直接采用紫外分光光度法测定PQP的含量误差较大。本课题旨在建立专属性强、准确、简便的定量分析方法,采用HPLC-UV法测定双氢青蒿素哌喹片中的PQP。伤湿祛痛膏是由川乌、草乌等药材制成的复方橡皮膏剂,主要用于头痛、风湿痛、神经痛等。因为川乌、草乌中均含有剧毒的乌头碱类酯型生物碱,而伤湿祛痛膏的我国现行国家药品质量标准中,并未对乌头碱进行限量控制。因此,为完善伤湿祛痛膏的质量标准,更好地控制产品质量,本课题采用薄层色谱法（Thin Layer Chromatography, TLC）对伤湿祛痛膏中乌头碱的限量检查进行研究,该检查方法简便、结果准确。方法:双氢青蒿素的含量测定采用HPLC法,用外标法以α和β体峰面积之和对DHA进行定量分析。理论板数按α异构体峰或β异构体峰计算均不低于10000。色谱条件色谱柱:Phenomenex C18柱（250 mm×4.6 mm,5μm）;流动相:乙腈-甲醇-0.02 mol·L^-1硫酸铵（50 : 10 : 40,稀硫酸调节至pH 4.8）;流速:1.0 mL·min^-1;检测波长:210 nm;柱温:30℃;进样量:10μL。磷酸哌喹的含量测定采用HPLC法,用外标法以峰面积对PQP进行定量计算。理论板数以PQP峰计不低于2000。色谱条件色谱柱:Phenomenex C18柱（250 mm×4.6 mm,5μm）;流动相:乙腈-0.022 mol·L^-1盐酸（30 : 70,三乙胺调节至pH 2.3）;流速:1.0 mL·min^-1;检测波长:225 nm;柱温:30℃;进样量:10μL。乌头碱的限量检查采用TLC对照品比较法,实现良好分离,重现性好。色谱条件展开剂:正己烷-乙酸乙酯-无水乙醇（6.4 : 3.6 : 1）;显色剂:稀碘化铋钾试液。结果:双氢青蒿素的含量测定方法准确度高,回收率为98.6%;精密度好,RSD为0.49%（n = 5）;定量限为80 ng（S/N = 10）;DHA在0.84 25.12μg的范围内,峰面积与浓度间呈现良好的线性,线性方程为A = 29987c + 1006.6,γ= 0.9999（n = 7）;供试品溶液在分析浓度下,8小时内基本稳定。磷酸哌喹的含量测定方法准确度高,回收率为101.3%;精密度好,RSD为0.41%（n = 6）;定量限为90 ng（S/N = 10）;PQP在0.20 1.21μg的范围内,峰面积与浓度间呈现良好的线性,线性方程为A = 525092c + 36071,γ= 0.9998（n = 5）;供试品溶液在分析浓度下,8小时内基本稳定。乌头碱的限量检查乌头碱的分离效果良好,斑点清晰圆整、无拖尾现象;以稀碘化铋钾试液作显色剂,显色稳定,利于乌头碱斑点的检出和判定。更多还原

【Abstract】 OBJECTIVE:Compound tablets of Dihydroartemisinin and Piperaquine Phosphate are a new type of antimalarial drug. In Chinese current drug specification, DHA was determined by UV specterphotometry after basic hydrolysis. PQP was assayed with UV specterphotometry directly. However, there were much analytical errors in the two determinations. This research focuses on establishing sensitive and accurate HPLC-UV methods for determinations of DHA and PQP in compound tablets.Shangshiqutong Plasters are compound plasters made of Chinese traditional medicine Chuanwu, Caowu and so on. The plasters are an acesodyne. Chuanwu and Caowu contain Aconitine. Whereas it is virulent, Aconitine in Shangshiqutong Plasters was not tested in Chinese current drug specification. To establish TLC method for testing Aconitine in the compound plasters was another objective of this research.METHOD:Determination of DHA in Compound tablets: A Phenomenex C18 column （250mm×4.6mm, 5μm） was used. Mobile phase was acetonitrile - methanol - 0.02mol·L^-1 ammonium sulfate （50 : 10 : 40）, adjusted to pH 4.8 with dilute sulphuric acid. Flow rate was 1.0 mL·min^-1, detection wavelength was 210 nm, column temperature was 30℃, and injection volume was 10μl. Theoretical plates forα-DHA orβ-DHA were no less than 10000.Determination of PQP in Compound tablets: A phenomenex C18 column （250mm×4.6mm, 5μm） was used. Mobile phase was acetonitrile - 0.022mol·L^-1 hydrochloric acid （30 : 70）, adjusted to pH 2.3 with triethylamine. Flow rate was 1.0 mL·min^-1, detection wavelength was 225 nm, column temperature was 30℃, and injection volume was 10μL. Theoretical plate for PQP was no less than 2000.Test of Aconitine in Shangshiqutong Plasters: Mobile phase was n - hexane - ethyl - acetate - anhydrous - alcohol （6.4 : 3.6 : 1）. Colorant was dilute bismuth potassium iodide.RESULTS:Determination of DHA in Compound tablets: Recovery was 98.6%. RSD of precision was 0.49% （ n = 5）. LOQ was 80 ng （S/N = 10）. Linear range was 0.84 25.12μg, linear regression equation was A = 29987c + 1006.6,γ= 0.9999 （n = 7）. Sample solutions were stable in 8 h.Determination of PQP in Compound tablets: Recovery was 101.3%. RSD of precision was 0.41% （n = 6）. LOQ was 90 ng （S/N = 10）. Linear ranges was 0.20 1.21μg, linear regression equation was A = 525092c + 36071,γ= 0.9998 （n = 5）. Sample solutions were stable in 8 h.Test of Aconitine in Shangshiqutong Plasters: The spot of Aconitine was separated from other spots completely. The colors of spots were stable.更多还原