HPLC-MS法测定大鼠血浆中广藿香酮衍生物PPCl浓度及其药动学研究

目的建立大鼠血浆中广藿香酮衍生物(PPCl)浓度的HPLC-MS分析方法,并研究该药在大鼠体内的药动学特征。方法采用乙酸乙酯萃取大鼠血浆中的药物和内标(大黄素),以Infinity Lab Poroshell 120 EC-C18(3mm×100mm,2.7μm)为色谱柱;以乙腈-水(5mmol/L醋酸铵)为流动相梯度洗脱,流速为0.3mL/min;质谱采用ESI离子源和负离子多重反应离子监测方式测定大鼠血浆中PPCl的浓度。结果大鼠血浆中的PPCl在33.26~30800ng/mL线性范围内线性关系良好(r=0.997);批内、批间精密度和准确度符合生物样品的检测要求;提取回收率和基质效应的平均值分别为99.83%和94.40%;大鼠灌胃给予10mg/kg的PPCl,药物的t1/2为(14.57±1.93)h,Cmax为(19.29±6.47)μg/mL,AUC0-t为(434.40±140.09)μg/(mL·h),结论本实验建立的测定大鼠血浆中PPCl的HPLC-MS法,操作简单,灵敏度高,专属性强,无内源性及其他杂质干扰,适用于大鼠血浆中PPCl浓度的测定及其药动学研究。

An HPLC-MS method for the determination of pogostone derivative PPCl in rat plasma and its pharmacokinetic profiles

Objective To establish a liquid chromatography-tandem mass spectrometry (HPLC-MS) method for the determination of pogostone derivative PPCl in rat plasma, and study the pharmacokinetic characteristics of PPCl by intragastric administration. Methods The PPCl and internal standard substance (emodin) were extracted from plasma samples using liquid-liquid extraction technique with ethyl acetate and chromatographed on Infinity Lab Poroshell 120 EC-C18 (3mm×100mm, 2.7μm) coupled with a guard column using acetonitrile-ammonium acetate (5mmol/L) as the mobile phase at a flow rate of 0.3mL/min. The MS detection was conducted by using HPLC-MS in a negative electrospray and multiple reactions monitor mode. Results Excellent liner relationship was obtained in the range of 33.26~30800ng/mL (r=0.997). The precision and the accuracy is according with the standard of the requirements for bioanalysis. The recovery and the matrix effect were respectively 99.83% and 94.40%. The rats were given 10mg/kg of PPCl by intragastric administration. The pharmacokinetic parameters of PPCl were as following: t1/2 (14.57±1.93)h, Cmax(19.29±6.47)μg/mL and AUC0~t(434.40±140.09)μg/(mL·h). Conclusion The HPLC-MS method is sample, highly sensitive, specific with no endogenous interference and it can be used for the determination of PPClin rat plasma and study the pharmacokinetic characteristics of PPCl in