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高效液相色谱电化学检测法分析大鼠血浆中的四种黄连生物碱。

Analysis of four alkaloids of Coptis chinensis in rat plasma by high performance liquid chromatography with electrochemical detection.

机构信息

Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Wuhan University, Ministry of Education, and Wuhan University School of Pharmaceutical Sciences, Wuhan, China.

出版信息

Anal Chim Acta. 2012 Aug 6;737:99-104. doi: 10.1016/j.aca.2012.05.054. Epub 2012 Jun 4.

Abstract

A sensitive and precise high performance liquid chromatography (HPLC)-electrochemical detection (ECD) method has been developed for the simultaneous determination of four isoquinoline alkaloids including berberine, jatrorrhizine, coptisine and palmatine in Chinese medicine Coptis chinensis. The typical HPLC analysis was performed on WondaSil(®) C18-WR column (250×4.6 mm, 5 μm) with the mobile phase comprising 40 mM phosphate buffer (pH 7.0)-acetonitrile (40:60, v/v) at the flow rate of 0.8 mL min(-1). The electrochemical detection employed a three electrode system with a bare glassy carbon electrode at +1.3 V versus the Ag/AgCl reference electrode. The limits of detection (LODs) of four alkaloids ranged from 0.01 to 0.03 μmol L(-1) and the LOD of berberine was 80 times lower than LOD obtained by UV detection. The rat plasma samples were assayed after oral administration of the traditional Chinese medicine Coptis chinensis by the proposed HPLC-ECD method. The recoveries of this method were ranging from 88.0 to 116%, with the relative standard deviation lower than 3.1% for intra-day precision and 5.7% for inter-day precision. These results show that HPLC-ECD is a useful tool for the quality control of herbal medicine Coptis chinensis and also for pharmacokinetic studies.

摘要

建立了一种灵敏、精确的高效液相色谱-电化学检测(HPLC-ECD)法,用于同时测定黄连中的 4 种异喹啉生物碱,包括小檗碱、黄连碱、小檗碱和巴马汀。典型的 HPLC 分析在 WondaSil(®)C18-WR 柱(250×4.6mm,5μm)上进行,流动相为 40mM 磷酸盐缓冲液(pH7.0)-乙腈(40:60,v/v),流速为 0.8mL min(-1)。电化学检测采用三电极系统,裸玻碳电极为+1.3V 相对于 Ag/AgCl 参比电极。4 种生物碱的检测限(LOD)范围为 0.01-0.03μmol L(-1),小檗碱的 LOD 比 UV 检测的 LOD 低 80 倍。采用所建立的 HPLC-ECD 法测定口服黄连后大鼠血浆样品。该方法的回收率在 88.0%至 116%之间,日内精密度的相对标准偏差低于 3.1%,日间精密度的相对标准偏差低于 5.7%。这些结果表明,HPLC-ECD 是一种用于黄连质量控制和药代动力学研究的有用工具。

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