反相高效液相色谱法测定体外人肝微粒体中氟西汀及其代谢产物去甲氟西汀

目的:建立同时测定体外肝微粒体中氟西汀及其代谢产物去甲氟西汀的反相高效液相色谱紫外检测法.方法:含微粒体蛋白和NADPH发生系统及氟西汀的孵育液加入冰乙腈酸终止反应后,再加入去甲替林作为内标并以 n-正己烷和乙腈的混合液进行萃取,然后以反相ODS柱分离,在226 nm处以紫外检测器进行检测.结果:孵育体系中没有明显的干扰峰出现,氟西汀和去甲氟西汀洗脱快,分离好,线性范围均为10-800μg/L,最低检测限均为5μg/L,相对回收率为94％-104％,变异系数少于9.1％.结论:本法快速,灵敏,回收率高,且萃取过程简单,可用于体外氟西汀的代谢研究.

Determination of fluoxetine and its metabolite norfluoxetine in human liver microsomes by reversed-phase HPLC in vitro

AIM: A high-performance liquid chromatography (HPLC) method was developed for the determination of fluoxetine (FLU) and its metabolite norfluoxetine (N-FLU) in human liver microsomes in vitro. METHODS: An incubation buffer containing human liver microsomes, NADPH-generating system, and FLU, after termination of enzyme reaction and addition of nortriptyline (NOR) as internal standard (IS), was extracted with n-hexane/acetonitrile, and separated on a reversed-phase ODS column. Detection was achieved at 226 nm by ultraviolet detector (UV). RESULTS: The limit of detection was 5 micrograms/L for both FLU and N-FLU. No potential interference was found. The method provides recoveries of up to 94%-104% and acceptable coefficients of variation were found for both within-run (< 7.8%) and day to day (< 9.1%) assays. CONCLUSION: This method is rapid, sensitive, and simple for studying the metabolism of FLU and N-FLU.