HPLC-MS/MS法测定人血浆中喹那普利及代谢产物喹那普利拉的浓度

目的:建立HPLC-MS/MS法测定人血浆中喹那普利及其代谢产物喹的浓度。方法:血浆经甲醇蛋白沉淀,使用Agilent C₁₈色谱柱(4.6 mm×150 mm,5 μm),以甲醇-0.3%甲酸水(60:40)为流动相,流速1 mL·min^-1,依那普利拉为内标;通过电喷雾离子化四极杆串联质谱,正离子多反应检测方式(MRM)检测,用于定量分析的离子反应分别为m/z 439.2→234.1(喹那普利)、m/z 411.1→206.1(喹那普利拉)、m/z 349.1→206.1(依那普利拉)。结果:喹那普利与喹那普利拉质量浓度分别为4.096~10 000 μg·L^-1和8.192~2 000 μg·L^-1时线性关系良好(r²=0.996 6和0.995 9),日内精密度≤6.5%,日间精密度RSD≤8.1%,回收率≥94.9%。喹那普利与喹那普利拉的T_max分别为(0.6±0.2)h和(1.5±0.4)h,C_max分别为(243.3±90.1)μg·L^-1和(1 008.8±298.4)μg·L^-1,t_1/2分别为(0.7±0.2)h和(2.3±0.3)h,AUC_0-t分别为(220.9±85.6)h·μg·L^-1和(3 312.4±967.7)h·μg·L^-1。结论:本法经方法学验证,可用于测定药物喹那普利的血药浓度并进行药代动力学研究。

Objective:To establish an HPLC-MS/MS method for simultaneous determination of quinapril and its active metabolite quinaprilat in human plasma.Methods:The plasma samples were pretreated by protein precipitation method.Separation was performed on an Agilent C₁₈column (4.6 mm×150 mm,5 μm)using an isocratic mobile phase system consisting of methanol-0.3% formic acid aqueous solution (60:40)at a flow rate of 1 mL·min^-1 with enalaprilat as internal standard.Analytes were determined by tandem mass spectrometry with electrospray positive ionization and multiple-reaction monitoring (MRM)mode.The monitoring ions were m/z 439.2→234.1 for quinapril,m/z 411.1→206.1 for quinaprilat and m/z 349.1→206.1 for enalaprilat.Results:The linear range of the calibration curve was 4.096-10 000 μg·L^-1 with r²=0.996 6 for quinapril and 8.192~2 000 μg·L^-1 with r² =0.995 9 for quinaprilat.The recoveries were not less than 94.9%.The intra- and inter-day precisions were not more than 6.5% and 8.1%,respectively.The main pharmacokinetic parameters of quinapril and quinaprilat were as follows:T_max (0.6±0.2)h and (1.5±0.4)h,C_max (243.3±90.1)μg·L^-1and (1 008.8±298.4)μg·L^-1,t_1/2 (0.7±0.2)h and(2.3±0.3)h,AUC_0-t (220.9±85.6)h·μg·L^-1and (3 312.4±967.7)h·μg·L^-1.Conclusion:The established method is proved to be useful in determination of plasma drug concentration of quinapril,which is applicable in the pharmacokinetic study of quinapril.