LC-MS/MS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的血药浓度

Pharmacokinetic Study of Apigenin, 3-Hydroxy-genkwanin and Genkwanin in Rats after Oral Administration of Extraction of Dephne Genkwa Sieb.et Zucc by LC-MS/MS

关键词:  芹菜素  3’-羟基芫花素  芫花素  液相色谱-质谱联用

Key words:  apigenin  3-hydroxy-genkwanin  genkwanin  LC-MS/MS

摘要:

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，5 μm)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·mL^-1；定量下限为 10 ng·mL^-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。

Abstract:

OBJECTIVE To develop a sensitive and rapid HPLC-MS/MS method for the simultaneous quantification of apigenin, 3-hydroxy-genkwanin and genkwanin in rat plasma after oral administration of extraction of Dephne genkwa Sieb.et Zucc. METHODS Esculin was used as the internal standard(IS), and samples spiked with analyzes and the IS were processed using liquid-liquid extraction. Then, the extract chromatographed on a Lichrosorb RP-C18(4.5 mm×150 mm, 5 μm) column with the mobile phase consisted of methanol-0.5% formic acid (80∶20). A tandem mass spectrometer equipped with an ESI source was used as the detector and was operated in the negative ion mode. Quantification was performed using multiple reaction monitoring (MRM). RESULTS The calibration curves of apigenin, 3-hydroxy-genkwanin and genkwanin were linear over the range of 10-2 000 ng·mL^-1, and the lower limit of quantification was 10 ng·mL^-1. The validated method was successfully used to study the pharmacokinetics of apigenin, 3-hydroxy-genkwanin and genkwanin in rats after oral administration of extraction of Dephne genkwa Sieb.et Zucc. CONCLUSION The established LC-MS/MS method is validated to specific, accurate and sensitive for quantitation and pharmacokinetic study of medicines including apigenin, 3-hydroxy-genkwanin and genkwanin.