HPLC-MS/MS法同时测定大鼠血浆中水飞蓟宾A和飞蓟宾B及药代动力学___

目的：建立大鼠血浆中水飞蓟宾A和水飞蓟宾B的测定法，并用于大鼠体内的药代动力学研究。方法：采用静脉给药方式，蛋白沉淀法快速制备样品；分析柱为Agilent Zorbax Eclipse XDB-C18（5 μm,2.1 mm×150 mm），流动相为甲醇:0.1%的甲酸水溶液（48:52，v/v）；质谱条件为ESI( - )电离方式，扫描方式为选择性反应监测（SRM），范围m/z481.1→m/z300.9（水飞蓟宾A和B）和m/z579.2→m/z271.1（内标）。结果：分析方法的线性范围水飞蓟宾A和B均为20～50000 ng/mL，样品的检出限均为20 ng/mL,日内和日间精密度均≤9.1%，准确度在±5.8%之内，方法的回收率均在90%以上。药代动力学研究结果表明水飞蓟宾A和B在大鼠体内的动力学过程均符合二室模型。对参数计算结果进行非参数检验，显示其在水飞蓟宾A和B间均无显著性差异，表明二者在大鼠体内有相似的药代动力学行为。结论：本方法简单、灵敏、稳定可靠，适合于水飞蓟宾、水飞蓟素及其制剂的药代动力学研究。

 Objective:A liquid chromatography-tandem mass spectrometry(LC–MS–MS) method was developed and validated for the quantification of silybin A and silybin B to study their pharmacokinetics in rat plasma. Method:Intravenous administration was used in this paper and a simplified protein precipitation with acetonitrile was employed for the rapid sample preparation;The separation was achieved on a Zorbax Eclipse XDB-C18 column(150 mm×2.1 mm ID，5 μm particle size), using methanol–water containing 0.1% formic acid (48:52, v/v) as the mobile phase. With negative electrospray ionization;The mass spectrometer was operated in selected reaction monitoring (SRM) mode using the transition m/z 481.1→300.9 for both silybin A and silybin B and m/z 579.2→271.1 for naringin,respectively. Result: Linear calibration curves were obtained in the concentration range of 20–50000 ng/ml with a lower limit of quantitation of 20 ng/mL for both silybin A and silybin B, respectively;The intra- and inter-day precision values were below 9.1% and accuracy was within ±5.8% at all three quality control levels, and the recovery of assay were above 90% for both silybin A and silybin B, respectively;Mean plasma concentration-time curves of both silybin A and silybin B were best fitted to two-compartment models in rat.The pharmacokinetic parameters between silybin A and B was no significant difference. It shows that there is a similar pharmacokinetic behavior between silybin A and B in rats. Conclusion: The validated method is simple,sensitive,selective and reproducible for the study of the pharmaeokineties of silybin,silymarin or their preparations