Zhang Shuangqing, Sun Licui, Huang Zhenwu
Wei Sheng Yan Jiu. 2015 Jan;44(1):91-4.
To establish an ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS) method for the simultaneous quantification of icairitin(ICT) and its major metabolite glucuronidated icaritin (GICT), and investigate metabolism of ICT in rats following a single intraperitoneal administration.
ICT, GICT and an internal standard coumestrol (CMT) were extracted from rat plasma using acetonitrile and separated on a C18 column using a gradient mobile phase of acetonitrile, water, ammonium formate and formic acid. In the negative electrospray ionization mode, selected reaction monitoring of the precursor-product ion transitions of m/z 367.1 --> 297.1 for ICT, 543.3 --> 367.1 for GICT and 267.0 - 211.1 for CMT was used for the quantification. Plasma was collected for the determination of ICT and GICT after rats were intraperitoneally administered with ICT at a single dose of 10 mg/kg.
The calibration curves were linear over the concentration range of 0.2 - 20 ng/ml for ICT and 2 -200 ng/ml for GICT with the respective lower limit of quantification at 0.2 and 2 ng/ml. The intra- and inter-day accuracy values fell in the range of 95.1 - 103.7%, and precisions were within 10.3%. Recovery and matrix effect were 89.1 - 92.4% and 93.2 - 104.2%, respectively.
The UHPLC-MS/MS method was specific and accurate, suitable for the metabolism study of ICT. ICT was rapidly metabolized to GICT in rats.
建立一种超高效液相色谱 - 串联质谱法（UHPLC - MS/MS），用于同时定量测定淫羊藿素（ICT）及其主要代谢产物葡萄糖醛酸化淫羊藿素（GICT），并研究单次腹腔注射后ICT在大鼠体内的代谢情况。
采用乙腈从大鼠血浆中提取ICT、GICT和内标物香豆雌酚（CMT），并在C18柱上进行分离，流动相为乙腈、水、甲酸铵和甲酸的梯度洗脱。在负离子电喷雾电离模式下，通过选择反应监测ICT的m/z 367.1 --> 297.1、GICT的m/z 543.3 --> 367.1和CMT的m/z 267.0 - 211.1的前体 - 产物离子转换进行定量分析。大鼠单次腹腔注射10 mg/kg ICT后，采集血浆测定ICT和GICT。
ICT的校准曲线在0.2 - 20 ng/ml浓度范围内呈线性，GICT的校准曲线在2 - 200 ng/ml浓度范围内呈线性，各自的定量下限分别为0.2和2 ng/ml。日内和日间准确度值在95.1 - 103.7%范围内，精密度在10.3%以内。回收率和基质效应分别为89.1 - 92.4%和93.2 - 104.2%。
UHPLC - MS/MS方法特异、准确，适用于ICT的代谢研究。ICT在大鼠体内迅速代谢为GICT。
