LC-MS/MS测定大鼠血浆中黄藤素含量及其药代动力学研究

Determination of fibrauretin concentrations in rat plasma by LC-MS/MS and its pharmacokinetics

目的建立大鼠血浆中黄藤素的LC-MS／MS定量方法，并利用此方法考察黄藤素在大鼠体内的药物动力学。方法色谱柱为Zorbax Eclipse XDB—C18（2．1mm×50mm，1．8μm），流动相为0．1％甲酸溶液-乙腈（70：30），采用多级反应模式（MRM）检测；通过大鼠眼底静脉取血，血浆经乙腈沉淀蛋白，取上清液进行LC-MS／MS分析，测定黄藤素血药浓度，通过DAS2．0计算其药动学参数。结果血浆中黄藤素在0．442～44．2ng／ml范围内呈现良好的线性关系，方法学精密度、回收率、稳定性等均符合要求；大鼠体内黄藤素药动学符合二室模型特征，灌胃给予40mg／kg黄藤素后，大鼠血浆中的黄藤素浓度在2．8h达峰值，t1／2为6h，最大血药浓度为19．8ng／ml。结论本研究建立的大鼠体内黄藤素测定方法灵敏度高、专一性好，可用于黄藤素的体内药物动力学研究，为黄藤素应用研究提供了有力支持。

Objective To establish a LC-MS/MS method to determine fibrauretin concentrations in rat blood and study the pharmacokinetics of fibrauretin in rat by the method. Methods Separation was performed by an Agilent Zorbax Eclipse XDB-Cis column （2.1 mm × 50 mm, 1.8 μm） and the mobile phase consisted of acetonitrile and 0.1% formic acid aqueous solution （30 ： 70）. The quantification analysis of fibrauretin was performed using multiple reaction monitoring （MRM） pattern. The plasma was analyzed after a simple protein precipitation procedure with acetonitrile. The blood sample was collected from retinal vein, and the concentration of fibrauretin in rat blood was analyzed by the established method, and the pharmacokinetics parameters were calculated by the DAS2.0 software. Results The linear range of fibrauretin in rat plasma was 0.442 - 44.2 ng/ml. The precision, extraction recovery and stability limit of this method met the requirements of pharmacokinetic study. The pharmacokinetics of fibrauretin in rats was fitted with two-compartment model. After oral administration of fibrauretin of 40 mg/kg, the pharmacokinetic parameters including the half time （t1/2） and the peak time and were 6 h and 2.8 h respectively, and the maximum plasma concentration was 19.8 ng/ml.Conclusions Established in this study in vivo determination method of high sensitivity and specificity, can be used for pharmacokinetic studies of fibrauretin in rats, which provides a strong support for its applied research.