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超高效液相色谱-电喷雾串联质谱法测定杜仲提取物口服给药后大鼠血浆中松脂醇葡萄糖苷和绿原酸的含量及药代动力学研究

UHPLC-ESI-MS/MS determination and pharmacokinetics of pinoresinol glucoside and chlorogenic acid in rat plasma after oral administration of Eucommia ulmoides Oliv extract.

作者信息

Gong Xiaojian, Luan Qingxiang, Zhou Xin, Zhao Yang, Zhao Chao

机构信息

Key Laboratory for Information System of Mountainous Areas and Protection of Ecological Enviroment, Guizhou Normal University, Guiyang, China.

Guizhou Engineering Laboratory for Quality Control and Evaluation Technology of Medicine, Guiyang, China.

出版信息

Biomed Chromatogr. 2017 Nov;31(11). doi: 10.1002/bmc.4008. Epub 2017 Jun 13.

Abstract

This study aimed to develop a specific UHPLC-ESI-MS/MS method for simultaneous determination and pharmacokinetics of pinoresinol glucoside and chlorogenic acid in rat plasma after oral administration of Eucommia ulmoides. The chromatographic separation was achieved on a Hypersil GOLD column with gradient elution by using a mixture of 0.1% formic acid aqueous solution and acetonitrile as the mobile phase at a flow rate of 200 μL/min. A tandem mass spectrometric detection was conducted using multiple-reaction monitoring via an electrospray ionization source in negative ionization mode. Samples were pre-treated by a single-step protein precipitation with acetonitrile, and bergenin was used as internal standard. After oral administration of 3 mL/kg E. ulmoides extract in rats, the maximum plasma concentrations of pinoresinol glucoside and chlorogenic acid were 57.44 and 61.04 ng/mL, respectively. The times to reach the maximum plasma concentration were 40.00 and 23.33 min for pinoresinol glucoside and chlorogenic acid, respectively. The intra- and inter-day precision (RSD) values for the two analytes were <2.46 and 5.15%, respectively, and the accuracy (RE) values ranged from -12.76 to 0.00. This is the first study on pharmacokinetics of bioactive compounds in rat plasma after oral administration of E. ulmoides extract.

摘要

本研究旨在建立一种超高效液相色谱-电喷雾串联质谱法(UHPLC-ESI-MS/MS),用于同时测定杜仲叶提取物灌胃大鼠血浆中松脂醇葡萄糖苷和绿原酸的含量及药代动力学。采用Hypersil GOLD色谱柱,以0.1%甲酸水溶液和乙腈为流动相进行梯度洗脱,流速为200 μL/min,实现色谱分离。采用电喷雾电离源,在负离子模式下通过多反应监测进行串联质谱检测。样品采用乙腈一步沉淀法进行预处理,以岩白菜素为内标。大鼠灌胃给予3 mL/kg杜仲叶提取物后,松脂醇葡萄糖苷和绿原酸的最大血浆浓度分别为57.44和61.04 ng/mL。松脂醇葡萄糖苷和绿原酸达到最大血浆浓度的时间分别为40.00和23.33 min。两种分析物的日内和日间精密度(RSD)值分别<2.46%和5.15%,准确度(RE)值范围为-12.76至0.00。这是首次关于杜仲叶提取物灌胃大鼠血浆中生物活性化合物药代动力学的研究。

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