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)  

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

作　　者：李相鸿[1] 孙华[1] 贾元威[2] 周理想[1] 王伟佳[1] 戴敏[1] 赵亚男[1] 谢海棠[1,2] 

机构地区：[1]皖南医学院弋矶山医院临床药学部,芜湖241001 [2]皖南医学院弋矶山医院安徽省药物临床评价中心,芜湖241001

出　　处：《天然产物研究与开发》2011年第6期1031-1037,共7页Natural Product Research and Development

基　　金：supported by the Youth Science Foundation of Wannan Medical College(No.WK200918F)

摘　　要：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。A high-performance liquid ehromatography/electrospray ionization tandem mass spectrometry （HPLC-ESIMS/MS） method was developed and validated for the determination of Bulleyaeonitine A （BLA） in human plasma using earbamazepinum 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,whieh was sensitive enough for the pharmacokinetic study of BLA. Ac- ceptable 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： C18 （0.325 ± 0.110）, （0.323 ± 0. 115 ） ng/mL; AUCo.,6 （ 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; T± （ 1.34 ± 0.54）, （1.83±0.99） h.

关 键 词：HPLC-MS/MS 草乌甲素 血药浓度 生物等效性 

分 类 号：R969.1[医药卫生—药理学]