Objective To establish an HPLC method for determining the concentration of cro-cetin in human liver microsomes.Methods After incubation with crocetin in the microsomal incu-bation system,the reaction was terminated and protein was precipitated by ice-cold acetonitrile. The HPLC analysis was performed with a reversed-phase Diamonsil C18 column (4.6×150 mm, 5 μm)at a column temperature of 30 ℃.The mobile phase consisted of methanol and 2% glacial acetic acid(7525,v/v)with a flow rate of 1.0 mL·min-1 .The detection wavelength was 440 nm.The indometacin was used as the internal standard.Results The linear range of this assay was 0.125-8 μg·mL-1 (R 2 =0.999 9).The extraction recoveries of low,medium and high concen-trations of crocetin were 97.24%,96.58% and 98.12%,respectively.The intra-and inter-batch relative standard deviation (RSD)was less than 15%,and the accuracy ranged from 85% to 115%.Conclusion The established HPLC method is simple,reliable and rapid for the detection of crocetin in human liver microsomes and meets the requirements for biological sample determina-tion.%目的：建立一种测定人肝微粒体孵育体系中藏花酸浓度的 HPLC 内标法。方法在人肝微粒体温孵体系中加入藏花酸样品进行孵育反应，反应结束后用冰乙腈终止反应。经沉淀法处理样品后，采用 Diamonsil C18色谱柱（4．6 mm×150 mm，5μm）进行分离。流动相：甲醇-2％冰乙酸水溶液（7525，v／v）；流速：1．0 mL·min－1；柱温：30℃；紫外检测波长：440 nm；内标：吲哚美辛。结果藏花酸经孵育反应后质量浓度测定方法的线性范围为0．125～8μg·mL－1，线性关系良好（R 2＝0．9999）。藏花酸低、中、高质量浓度的提取回收率分别为：97．24％、96．58％、98．12％，其 RSD 均小于15％，批内、批间差异＜15％，准确度为85％～115％。结论该 HPLC 法简便、准确，符合生物样品测定的要求，能快速、可靠地检测人肝微粒体中藏花酸的浓度。
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