HPLC-MS/MS法测定人血浆中奥昔布宁和去乙基奥昔布宁的质量浓度

HPLC-MS/MS determination of oxybutynin and N-desethyloxybutynin concentration in human plasma

目的建立HPLC-MS/MS法测定人血浆中奥昔布宁和去乙基奥昔布宁的质量浓度。方法选用Waters-At-lantis dC18色谱柱,以0.01%甲酸溶液-乙腈为流动相进行梯度洗脱,采用正离子多反应监测方式测定样品质量浓度。用于定量分析的离子对分别为m/z 358.2→142.2(奥昔布宁),m/z330.2→96.2(去乙基奥昔布宁),m/z268.2→152.2(奥昔布宁D10),m/z335.2→101.2(去乙基奥昔布宁D5)。结果血浆样品中,奥昔布宁在0.05～25ng.mL-1线性关系良好(r=0.996 5),最低定量浓度为0.05ng.mL-1;去乙基奥昔布宁在0.05～25ng.mL-1线性关系良好(r=0.998 5),最低定量浓度为0.05ng.mL-1。两者日内与日间RSD均<15%,提取回收率>75%,且稳定性均较好。结论本方法简便快速、灵敏准确、特异性强,适用于奥昔布宁和去乙基奥昔布宁的体内药物动力学研究。

Objective To establish a high performance liquid chromatography-mass spectrometry method for the determination of oxybutynin and N-desethyloxybutynin in human plasma.Methods Waters-Atlantis dC18 column was used with 0.01% formic acid solution-acetonitrile as the mobile phase in gradient elution.The multiple reaction monitor with positive mode was used.The transitions of ＋ m/z 358.2→142.2（oxybutynin）,＋ m/z 330.2→96.2（N-desethyloxybutynin）,m/z 268.2→152.2（oxybutynin-D10）and ＋ m/z 335.2→101.2（N-desethyloxybutynin-D5） were used for the quantitative analysis.Results The calibration curve for oxybutynin was linear at 0.05-25 ng·mL-1（r=0.996 5）,and the minimum quantitative concentration was 0.05 ng·mL-1.The calibration curve for N-desethyloxybutynin was linear at 0.05-25 ng·mL-1（r=0.998 5）,and the minimum quantitative concentration was 0.05 ng·mL-1.The inter-day and intra-day precisions were less than 15%,the average recovery was above 75%.Conclusion The method is simple,rapid,sensitive,and accurate for the pharmacokinetic study of oxybutynin and N-desethyloxybutynin in human body.