肝细胞萃取-HPLC分析法的建立及其在栀子 保肝效应物质初步分析中的应用

目的:建立肝细胞萃取-HPLC分析法(HE-HPLC),并应用其分析栀子中保肝作用的活性成分.方法:应用肝细胞选择性地结合中药提取液中的活性成分,洗去未结合的其他成分后,使细胞靶点失活,被结合的活性成分从肝细胞中释放出来,用高效液相色谱法对这些成分进行分析.结果:可检测到栀子水提液的主要保肝效应成分,初步确定为2个,其中1个为栀子昔,另1个有待进一步鉴定.结论:肝细胞萃取-HPLC 分析法基本可以反映化合物与细胞生物靶点(受体、通道、酶等)的相互作用;该体系中的保留成分和其药理作用有显著的相关性,可用于中药复杂体系物质基础的分析.

Establishment of hepatocyte extraction combined with HPLC(HE-HPLC) and application in analysis of active components in the fruits of Gardenia jasminoides extract

To screen effective principles from traditional Chinese medicine, a method named hepatocyte extraction combined with HPLC (HE-HPLC) was establish in this study. The active principles in the fruits of Gardenia jasminoides Ellis extract were combined with the hepatocytes under imitated physiological environments. Then the unattached substances were washed off by PBS with pH 7.4. After that the conjugated compounds were eluted by PBS with pH 4.0. These compounds released from target sites were collected and handled through SPE to be condensed, and analyzed by HPLC. The results indicated that two characteristic active compounds in the fruits of G. jasminoides extract binded to the hepatocytes. One of them is geniposide. The other is continued to be identified. It is showed that active principles which could bind to hepotocyte (through receptors, Channels, enzymes, etc.) could be detected, at least partly, by HE-HPLC analysis. There was a significant correlation between the retention properties of the active compounds which was obtained by HE-HPLC and their pharmacological effects on hepotocytes.