HPLC-MS/MS法测定人血浆中草乌甲素的浓度及生物等效性研究(英文)

本文建立了一种快速、高灵敏的HPLC-MS/MS法用于检测人血浆中的草乌甲素浓度。血浆样品采用沃特斯HLB小柱进行固相萃取,汉邦C18色谱柱(150 mm×4.6 mm,5μm)进行分离,流动相为甲醇∶水(85∶15,v/v),水相含10 mmol/L的醋酸铵和0.1%的甲酸。采用ESI源和多反应监测(MRM)的方式进行检测,草乌甲素及内标的反应离子对分别为644.4/584.4和237.2/194.2,草乌甲素血药浓度在0.010～1.0 ng/mL范围内线性关系良好,最低定量限为0.010 ng/mL可以满足口服0.4 mg草乌甲素后血药浓度的检测,日内日间及质控样品精密度及准确度均在允许范围内。本检测方法被成功的应用在中国健康志愿者生物等效性研究中,20名志愿者口服0.4 mg草乌甲素试验制剂和参比制剂后主要药代动力学参数分别如下:Cmax(0.325±0.110),(0.323±0.115)ng/mL;AUC0-16(1.627±0.489),(1.732±0.556)ng.h/mL;AUC0-∞(1.730±0.498),(1.831±0.562)ng.h/mL;t1/2(4.26±0.95),(3.80±0.90)h;Tmax(1.34±0.54),(1.83±0.99)h。

Determination of Bulleyaconitine A in Human Plasma by HPLC-MS/MS and Its Bioequivalence Study

A high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) method was developed and validated for the determination of Bulleyaconitine A(BLA) in human plasma using carbamazepinum as internal standard(IS).Plasma samples were extracted with Waters Oasis HLB solid-phase cartridges,separated on a reversed C18 column(150 mm × 4.6 mm,5 μm) with a mobile phase of methonal-10 mmol/L ammonium acetate solution containing 0.1% formic acid(85∶15,v/v).Bulleyaconitine A and IS were detected in the multiple reaction monitoring mode(MRM) with precursor to product ion transitions of m/z 644.4/584.4 and 237.2/194.2,respectively.The method exhibited a linear range of 0.010-1.0 ng/mL for BLA in human plasma.The lowest limit of quantification(LLOQ) was 0.010 ng/mL,which was sensitive enough for the pharmacokinetic study of BLA.Acceptable precision and accuracy were obtained for concentrations of the calibration standard and the quality control(QC).The validated method was successfully applied for evaluation of a bioequivalence study in Chinese healthy volunteers.The main pharmacokinetics parameters after oral administration of 0.4 mg BLA test or reference formulation were as follows∶Cmax(0.325±0.110),(0.323±0.115) ng/mL;AUC0-16(1.627±0.489),(1.732±0.556) ng·h/mL;AUC0-∞(1.730±0.498),(1.831±0.562) ng·h/mL;t1/2 (4.26±0.95),(3.80±0.90) h;Tmax(1.34±0.54),(1.83±0.99) h.