液相色谱-串联质谱法测定大鼠血浆中forsklin的浓度及其药代动力学

摘要

Objective To develop a sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the determination of forsklin in rat plasma.Methods After extraction with methyl tert-butyl ether,chromatographic separation was performed on a C18 column with the mobile phase consisting of water ( 0.1％ formic acid)-acetonitrile in a gradient elution mode.A tandem mass spectrometer equipped with electrospray ionization (ESI) source was used as detector in the positive ion mode.Quantification was performed using multiple reaction monitoring (MRM) with the precursor product combination ions of m/z 411→375.3 and 285→193 for forsklin and diazepam.Results Good linearity was obtained in the 0.5-1000 ng/ml range for the analyte and the analytical method was validated in terms of specificity,precision,accuracy,recovery,stability and matrix effect.These assays gave RSD values always lower than 14.4％ and RE values between -3.5 ％ and 3.8％.In addition,the specificity,extraction recovery,stability and matrix effect were satisfactory.Conclusion Due to its high sensitivity,specificity and simplicity,the method could be used for pharmacokinetic studies of forsklin.%目的 建立了测定大鼠血浆中forsklin的液相色谱-串联质谱法(LC-MS/MS).方法 血浆样品经叔丁基甲醚萃取后,以水(0.1％甲酸)-乙腈为流动相梯度洗脱,BetaBasic-C18柱分离.通过电喷雾离子化四极杆串联质谱,以多反应监测(MRM)方式进行正离子检测,用于定量分析的离子对分别为m/z 411→375.3(forsklin)和285→193(地西泮).结果 在本实验条件下,forsklin血浆浓度测定方法的线性范围为0.5～1000 ng/ml,定量下限为0.5 ng/ml.日内、日间精密度(相对标准差RSD)均小于14.4％；准确度(相对误差RE)在-3.5％和3.8％之间.方法的专属性,绝对回收率,稳定性和基质效应均符合要求.结论 该法快速、灵敏、准确,可用于forsklin的药代动力学研究.