HPLC-MS测定大鼠体内戊乙奎醚血浆浓度

HPLC-MS to Determine Plasma Concentrations of Penehyclidine in Rat

目的:建立HPLC-MS测定大鼠体内戊乙奎醚血浆浓度。方法:以罗通定为内标,样品经碱化后分别以3 mL重蒸乙醚萃取两次。分析柱Agilent Zorbax SB-C18(50×2.1 mm,3.5μm),柱温40℃;流动相为CH3CN∶5 mmol L-1NH4Ac∶0.2%HCOOH(50∶47∶3),流速0.2mL min-1,采用ESI离子源,以SIR正离子方式进行检测,检测离子为m/z 316.30(戊乙奎醚),m/z 356.22(内标)。结果:pH在0.205～250μg L-1范围内线性关系良好(r=0.9990),最低定量限为0.205μg L-1。日内、日间精密度均小于5%,方法学回收率为95.12%～105.8%。结论:本方法快速、准确、灵敏、选择性高,适用于戊乙奎醚在动物体内的药动学研究。

OBJECTIVE： To develop a HPLC-MS method for the quantification of penehyclidine in rat plasma.METHODS： Rotundine was used as internal standard（IS）,rat plasma samples were alkalified and extracted with ethylether,pH and IS were separated on a C18 column with a mobile phase containing CH3CN∶5 mmol L-1 NH4Ac∶0.2 % HCOOH（50∶47∶3） at a flow rate of 0.2 mL min-1.The electrospray ion trap mass spectrum in the positive mode and SIR were adopted to detect the concentration of penehyclidine.RESULTS Calibration curves were linear（r=0.9990） from 0.205 ~250 μg L-1.Both intra-and inter-batch precisions were less than 5 %.Methodology recovery was 95.12 %~105.80 %.Limit of quantification for penehyclidine was 0.205 μg L-1.CONCLUSION： HPLC-MS method described herein had been successfully applied for rat pharmacokinetic study,which offers advantages of specificity,quickness and higher sensitivity over the previously reported methods.