HPLC-MS法同时测定大鼠血浆中苦参碱、氧化苦参碱和氧化槐果碱的浓度及其药代动力学

Simultaneous determination of matrine,oxysophocarpin and oxymatrine in rat plasma by HPLC-MS and its application in the pharmacokinetic study

建立同时测定大鼠灌胃给予三物黄芩汤后血浆中苦参碱、氧化苦参碱和氧化槐果碱含量的HPLC-MS分析方法,并计算了3种生物碱在大鼠体内的药代动力学参数。血浆样品经氯仿液-液萃取后,以乙腈-0.1%甲酸水溶液(10∶90)为流动相,用KromasilC18色谱柱(4.6mm×150mm,5μm)分离,采用电喷雾离子化源(ESI)单重四极杆串联质谱,以选择离子检测(SIM)方式进行检测。苦参碱、氧化苦参碱和氧化槐果碱分别在10～5000ng.mL-1、2～1000ng.mL-1和2～1000ng.mL-1呈良好线性关系,提取回收率分别为89.1%～93.5%、83.9%～91.3%、85.4%～88.0%。日内及日间精密度均<15%。该法快速,灵敏,专属,适用于同时测定生物样本中苦参碱、氧化苦参碱和氧化槐果碱的血药浓度。

To establish an HPLC-MS method for simultaneous determination of matrine, oxymatrine and oxysophocarpine in rat plasma after oral administration of herbal preparation, namely Sanwu Huangqin decoction, and the pharmacokinetic porameters were calculated as well. Matrine, oxymatrine, oxysophocarpine, and internal standard pseudoephedrine were extracted from plasma with liquid-liquid extraction, then separated on a Kromasil C18 column by using acetonitrile-0. 1% aqueous formic acid （10：90） as mobile phase. Electrospray ionization （ESI） source was applied and operated in positive ion mode. The linear calibration curve was obtained in the concentration range of 10 - 5 000 ng · mL^-1 for matrine, 2 - 1 000 ng · mL^-1 for oxymatrine, and 2 - 1 000 ng · mL^-1 for oxysophocarpine. The extraction recovery was 89. 1% - 93.5% , 83.9% - 91.3% , and 85.4% - 88.0% accordingly. The inter- and intra- day precision （RSD） was below 15.0% calculated from quality control （QC） samples. Matrine, oxymatrine and oxysophocarpine concentration time profile conformed to a two-compartment pharmacokinetic model. The method was shown to be effective, convenient, and suitable for simultaneous pharmacokinetic study of matrine, oxymatrine, and oxysophocarpine in rat.