LC-MS/MS测定大鼠血浆中liguzinediol浓度及药动学

An LC-MS/MS method for determining liguzinediol in rat plasma and studying its pharmacokinetics

目的:建立灵敏、快速测定大鼠血浆中liguzinediol浓度的LC-MS/MS方法,探讨liguzinediol在雌雄大鼠体内的药动学。方法:血浆样品加入内标(咖啡因),经甲醇沉淀蛋白后,高速离心,上清液使用0.22μm微孔滤膜过滤,进行LC-MS/MS分析。采用Shim-pack XR-ODS色谱柱(50 mm×2.0 mm,2.2μm),流动相为甲醇-0.1%甲酸梯度洗脱,流速0.4 mL.min-1,柱温40℃。质谱采用ESI离子源,多反应监测模式(MRM)进行正离子检测,liguzinediol和内标咖啡因的选择性检测离子分别为m/z 169.2→122.2和m/z195.2→110.2。SD大鼠随机分为12组(雌雄各6组),每组6只,分别以灌胃和静脉注射的方式给予liguzinediol 5,10,20 mg.kg-1,测定给药后不同时间的血药浓度,利用DAS软件计算药动学参数。结果:liguzinediol的血药浓度在10～20 000 ng.mL-1范围内线性关系良好(r=0.999 6)。方法学考察均符合要求,日内、日间精密度RSD均<15%,符合生物样品分析要求。药动学研究表明雌雄大鼠间药动学参数有明显差异。结论:该方法操作简便、灵敏、快速、专属性强,可用于liguzinediol在大鼠体内的药动学及成药性研究。

Objective： To establish a simple and sensitive LC-MS/MS method for determining liguzinediol and study the pharmacokinetics of liguzinediol in male and female rats, respectively. Methods： Measurement of liguzinediol was performed by positive ion electrospray ionization in multiple reaction monitoring mode, and monitoring the transitions m/z 169.2→H22.2 for liguzinediol and m/z 195.2→110.2 for caffeine, respectively. Separation of liguzinediol and caffeine in plasma sample was carried out on Shim-pack XR-ODS column （50 mm ×2.0 mm, 2.2 μm） eluting with a gradient mobile phase system at a flow rate of 0.4 mL·min^-1. The sample injection volume was 2 μL and the column temperature was maintained at 40 ℃. Seventy-two Sprague-Dawley rats were divided into 6 male groups and 6 female groups. Three male groups and three female groups were administrated with an oral dose of 5, 10 and 20 mg· kg^- 1, respectively. Another three male groups and three female groups were administrated with an intravenous dose of S, 10 and 20 mg·kg^-1, respectively. The pharmacokinetic parameters were calculated by DAS 2.1 software. Results. Good linearity was obtained over the the correlation coefficient was better （ r = 0. 999 6）. They met the requirements range of 10 - 20 000 ng· mL^ -1, and of the analysis of biological samples with RSD of the intra- and inter-day precisions （ 〈 15% ）. Disparity was found between male and female rats in the pharmacokinetic parameters. Conclusion： The developed method is successfully applied to the pharmacokinetic study in rats.