大鼠肝微粒体中芬氟拉明对映体氧化代谢的立体选择性研究  

Study on stereoselectivity of fenfluramine enantiomers in rat liver microsome

作　　者：茅海琼[1] 曾苏[1] 

机构地区：[1]浙江大学药学院

出　　处：《中国现代应用药学》2005年第6期439-441,共3页Chinese Journal of Modern Applied Pharmacy

基　　金：国家自然科学基金(30225047)资助

摘　　要：目的建立大鼠肝微粒体孵育液中l-和d-芬氟拉明的对映体选择性测定的方法,并用于研究芬氟拉明Ⅰ相代谢的立体选择性.方法将dl-芬氟拉明与大鼠肝微粒体孵育后,采用柱前衍生化毛细管气相色谱-氢火焰离子化检测法进行对映体的分离,测定时间反应曲线和酶动力学参数.结果单个对映体的线性范围是1～50μg/mL;l-和d-芬氟拉明的方法平均回收率分别为92.4%和95.5%,检测限和定量限分别为0.1μg/mL和1.0μg/mL,方法精密度为RSD＜10%(n=6).l-和d-芬氟拉明的Km,,Vmax,Clint值分别为(0.15±0.01)和(0.27±0.02)μmol·mL-1,(4.99±0.52)和(9.53±0.87)nmol·g-1·min-1,(33.3±3.0)和(35.3±3.1)mL·min-1·g(蛋白)-1.结论方法准确可靠,已用于l-和d-芬氟拉明在大鼠肝微粒体孵育液中的代谢及其动力学研究;结果表明,芬氟拉明在大鼠的Ⅰ相代谢具有对映体选择性.OBJECTIVE To develop an enantioselective assay method for 1- and d- fenfluramine and study the stereoseleetivity of the phase Ⅰ metabolism of fenfluramlne enantiomers in rat liver mierosome. METHODS Separation and quantitation of the diastereomerle fenfluramine derivatives were carried out by a capillary gas chromatographic system with flame ionization detection. The time course and the parameters of enzymatic kinetics of fenfluramine enantiomers in the phase I metabolism were determined. RESULTS The assay was linear from 1 to 50μg/mL for each enantiomer. The analytical method affords average recoveries of 92.4% and 95.5% for 1- and d- fenfluramine, respectively. The limits of detection and quantitation of the assay were 0.1 and 1.0μg/mL for 1- and d-fenfluramine, respectively. The reproducibility of the assay was RSD 〈 10% （ n = 6）. Km, Vmax, C1 nt measured were （ 0.15± 0.01 ） and （0.27±0.02）1xmol· mL^-1, （4.99±0.52）and （9.53±0.87）nmol· g^-1· min^-1, （33.3±3.0） and （35.3±3.1） mL·min^-1 ·g（protein）^ -1 for 1- and d- fenfluramine, respectively. CONCLUSION The method allowed study of the depletion of 1- and d- fenfluramine in rat liver microsomal incubates. The stereoselectivity of fenfluramine phase Ⅰ metabolism was observed.

关 键 词：芬氟拉明 对映体 肝微粒体 代谢 

分 类 号：R917.7[医药卫生—药物分析学]