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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.
OBJECTIVE: To establish an RP-HPLC method for the simultaneous determination of fluoxetine and its metabolite nf-fluoxetine in liver microsomes.Methods: Incubation system containing microsomal protein and NADPH and fluoxetine After addition of glacial acetic acid to terminate the reaction, nor nortriptyline was added as an internal standard and extracted with a mixture of n-hexane and acetonitrile, followed by reverse phase ODS separation and detection by UV detector at 226 nm. There was no obvious interference peak in the incubation system. Flucloxacin and norfloxacin eluted quickly and separated well. The linear range was 10-800μg / L, the lowest detection limit was 5μg / L, the relative recovery was 94% -104%, coefficient of variation of less than 9.1% .Conclusion: The method is rapid, sensitive, high recovery rate, and the extraction process is simple and can be used for the study of fluoxetine metabolism in vitro.