LC-MS/MS法同时测定大鼠血浆中贝母素甲、贝母素乙的浓度及其在药代动力学中的应用

Simultaneous determination of peimine and peiminine in rat plasma by LC-MS/MS and its application in the pharmacokinetic study

建立同时测定大鼠灌胃及静注给予浙贝母提取物后血浆中贝母素甲及贝母素乙含量的HPLC-MS/MS分析方法,并计算2种生物碱在大鼠体内的药代动力学参数。血浆样品经乙酸乙酯液-液萃取后,以乙腈-10mmol·L-1甲酸铵水溶液(35∶65)为流动相,Luna C18色谱柱(150mm×2mmID,3μm)分离,采用电喷雾离子化源(ESI)三重四极杆串联质谱,以多反应监测模式(MRM)进行检测,检测离子对分别为m/z432.4→414.4(贝母素甲)、m/z430.4→412.4(贝母素乙)和m/z237.1→194.2(内标,卡马西平)。贝母素甲及贝母素乙均在0.8～800ng·mL-1内呈良好线性关系,提取回收率分别为94.1%～105.3%和85.8%～98.6%。方法的精密度、准确度和稳定性均符合要求。结果显示,该法选择性强、灵敏度高、操作简便,适用于贝母素甲、贝母素乙的大鼠药代动力学研究。

To establish an LC-MS/MS method for simultaneous determination of peimine and peiminine in rat plasma after oral and intravenous administration of Fritillaria thunbergii Miq.extract,the pharmacokinetic parameters were calculated as well.Peimine,peiminine and internal standard carbamazepine were extracted from plasma with liquid-liquid extraction by ethyl acetate,then separated on a Luna Cl8 column by using acetonitrile-water containing 10 mmol·L-1 ammonium formate（35：65）,as mobile phase.The electrospray ionization（ESI） source was applied and operated in positive ion mode.Peimine was detected at m/z 432.4 → 414.4,peiminine at m/z 430.4 → 412.4 and carbamazepine（IS） at 237.1 → 194.2.The linear calibration curves were obtained at the concentration range of 0.8-800 ng·mL-1 for peimine and peiminine.The extraction recoveries were 94.1%-105.3% and 85.8%-98.6%,respectively.The precisions,accuracy and stability of the analytes meet the requirements.The method was shown to be effective,convenient,and suitable for simultaneous pharmacokinetic study of peimine and peiminine in rat.