硝苯吡啶的HPLC测定方法及在人肝微粒体内的代谢动力学

HIGH-PERFORMANCE LIQUID CHROMATOGRATHY ASSAY FOR NIFEDIPINE IN BIOLOGICAL FLUIDS AND KINETICS OF NIFEDIPINE METABOLISM BY HUMAN LIVER MICROSOMES

本文建立了测定生物样品中硝苯吡啶浓度的反相HPLC法.选用非那西丁作为内标物,固定相用Nova-Pak C18分析柱,流动相为甲醇-水(68:32),紫外检测波长为240nm.样品由乙酸乙酯一次性提取,硝苯吡啶回收率为95.3±4.0%.本方法最低检测限为4ng.硝苯吡啶在人肝微粒体内代谢需NADPH再生系统,生成一个主要代谢物;按原药消除计算,硝苯吡啶代谢的Km和Vmax分别为58.0μmol/L和1.

A high-performance liquid chromatographic method was developed to determine nifedipine in biological fluids, by using ultraviolet detection at 240 nm.Phenacetin was chosen as internal standard.A Nova-Pak C18(4 um, 3.9×150mm) was employed with methanol-water (68:32) as mobile phase. Nifedipine was extracted by ethyl acetate with a percentage of recovery about 95.3±1.0%. The limit of detection of this method for nifedipine was 4 ng for an injection volume of 20 ul. One major metabolite was formed from nifedipine after incubation with human liver microsom-es, the NADPH generating system was needed in this metabolic ptocess. Km and Vmax measured by nifedipine disappearance from roicrosomal incubation mixture were 58.0 umol/L and 1.51 nmol/mg/min, respectively