LC-MS法测定离体大鼠肝灌流模型中知母皂苷B2的含量及药动学研究  

Determination of Timosaponin B2 in K -H Buffer by LC -MS and Pharmacokinetics of Timosaponin B2 in Isolated Perfused Rat Liver Model

作　　者：尤杰[1,2] 赫蕾[2] 陈明苍[2] 季宇彬[1] 潘国宇[2] 

机构地区：[1]哈尔滨商业大学生命科学与环境科学研究中心,黑龙江哈尔滨150076 [2]中国科学院上海药物研究所,上海201203

出　　处：《中药新药与临床药理》2013年第1期70-74,共5页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家科技重大专项<重大新药创制>课题(2012ZX09301001-001)

摘　　要：目的建立简单、快速的测定K—H缓冲液中知母皂苷B2含量的LC—MS方法，检验特定剂量知母皂苷B2在大鼠离体肝灌流模型中的肝脏提取率。方法将SD大鼠离体肝灌流给予知母皂苷B2，取出不同时间点肝灌流液，LC—MS法测定肝灌流液中知母皂苷B2的浓度。色谱条件：ACEC18色谱柱（100mm×2．1mm，1μm），流动相为乙腈-0．05％甲酸水溶液（V／V），采用梯度洗脱。结果知母皂苷B2在0．125—2μg／mL范围内线性良好（r2=0．999），日间和日内变异系数（RSD）〈10％，基质效应RSD〈15％，符合生物样品分析的方法学要求。结论该方法准确、灵敏、快速、高效，适用于知母皂苷B2在离体灌流液中的定量分析。获得了1．6μg／mL的知母皂苷B2样品大鼠离体肝灌流状态下的肝提取率和肝清除率曲线。Objective To develop a LC-MS method to determine timosaponin B2 in K-H buffer quickly and ef- fectively, and by using this method to study the metabolism of timosaponin B2 in isolated perfused rat liver model. Methods The isolated SD rat liver was peffused with K-H buffer containing timosaponin B2, and then the perfusate was taken out at different time for the determination of timosaponin B2 concentration by LC-MS method. The chromato- graphic condition was as follows： ACE C18 column （ 100 mm×2.1 mm, 1μm）, mobile phase consisting of acetonitrile -0.05 % formic acid V/V） , gradient elution. Results The linear ranges of timosaponin B2 was 0. 125 -2μg/mL （r2=0.999）. The intra-day and inter-day coefficient of differentiation was less than 10%, and the coefficient of dif- ferentiation for matrix effect was less than 15 %, which fit the standard. Conclusion The method is effective, rapid and accurate for tile determination of timosaponin B2 in K-H buffer, and the hepatic extraction rate and a liver clear- ance curve of timosanpnin B2 at 1.6μg/mL have beengot in isolated perfused rat liver.

关 键 词：知母皂苷B2 液质联用 方法学 基质效应 肝灌流 

分 类 号：R285.5[医药卫生—中药学]