HPLC-MS/MS法测定人血浆中己烯雌酚的浓度

摘要

A sensitive,fast,and reproducible high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS)method for the determination of diethylstilbestrol in human plasma was developed and validated.The plasma samples were pretreated by direct deproteinization with ethyl acetate.Daidzein was used as the internal standard.The separation was carried out on a Agilent Technologies 1200 series XDB C18 column (2.1 mm× 150 mm,5 μm) with a mobile phase of acetonitrile-2.5 mmol/L ammonium acetate (60∶40,v/v).Triple quadrupole mass spectrometric detection in negative ion mode was used for multiple-reaction-monitoring of the transitions at m/z 267.2→237.3 and m/z 253.2→132.3 for diethylstilbestrol and daidzein,respectively.The calibration curves were linear over the concentration range from 0.1 to 20 ng/mL (r2 =0.9984).The lower limit of quantification was 0.1 ng/mL (s mLs) for diethylstilbestrol,which was sensitive enough to perform pharmacokinetic studies after diethylstilbestrol administration.Inter-day and intra-day precisions were no more than 7％ with accuracies of 90％-105％.This method could be applied to therapeutic drug monitoring of diethylstilbestrol,which is helpful for evaluating the clinical efficacy and safety of diethylstilbestrol.%建立灵敏、高效、可重复的HPLC-MS/MS法测定人血浆中己烯雌酚的浓度.血浆经乙酸乙酯蛋白沉淀,黄豆苷元为内标,使用Agilent C18色谱柱(4.6 mm× 150 mm,5μm),以乙腈-2.5 mM醋酸铵水溶液(60∶40,v/v)为流动相,通过电喷雾离子化四极杆串联质谱,负离子多反应检测模式(MRM),用于定量的离子反应分别为m/z 267.2→237.3(己烯雌酚)、m/z 253.2→132.3(黄豆苷元).己烯雌酚浓度为0.1-20 ng/mL时线性关系良好(r2=0.9984),该方法的重复性实验RSD为4.7％,准确度为90％-105％,最低检测限为0.1 ng/mL.该方法重现性好,定量限低,适用于检测药物己烯雌酚的血药浓度并进行药代动力学研究.