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目的建立一个灵敏、快速的液相色谱-质谱联用法同时测定大鼠口服芫花粗提物后芹菜素、3’-羟基芫花素和芫花素在血浆样品中的浓度。方法以秦皮甲素为内标,大鼠血浆样品经液液萃取后,以甲醇-0.5%甲酸(80∶20)为流动相,采用Lichrosorb Rp-C18(4.5 mm×150 mm,5μm)色谱柱分离,通过ESI三重四级杆串联质谱,以多反应监测(MRM)方式进行检测。结果芹菜素、3’-羟基芫花素和芫花素测定方法的线性范围均为10~2 000 ng·m L-1;定量下限为10 ng·m L-1。该方法被成功应用于芫花粗提物在大鼠体内的药动学研究。结论该方法灵敏度高,操作简便,分析速度快,适用于含有芹菜素、3’-羟基芫花素或芫花素的药物测定或药动学研究。
Objective To establish a sensitive and rapid LC-MS method for the simultaneous determination of apigenin, 3’-hydroxy genkwanin and genkwanin in rat plasma samples after oral administration of Daphne genkwa. Methods Aesculin was used as an internal standard. The plasma samples of rats were extracted by liquid-liquid extraction using methanol-0.5% formic acid (80:20) as mobile phase. The chromatographic column was Lichrosorb Rp-C18 (4.5 mm × 150 mm, 5 μm) Separated, and detected by multiple reaction monitoring (MRM) by ESI triple quadrupole tandem mass spectrometry. Results The calibration curve of apigenin, 3’-hydroxy genkwanin and genkwanin showed a linear range of 10-2000 ng · m L-1 and a lower limit of quantitation of 10 ng · m L-1. The method was successfully applied to the pharmacokinetics of crude extract of Daphne genkwa in rats. Conclusion The method is sensitive, simple, and fast in analysis. It is suitable for the determination of pharmacokinetics or pharmacokinetics of apigenin, 3’-hydroxy genkwanin or genkwanin.