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

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-Atlantis 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／z358.2→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/z 335.2→101.2(去乙基奥昔布宁D5).结果 血浆样品中,奥昔布宁在0.05～25 ng·mL-1线性关系良好(r=0.996 5),最低定量浓度为0.05 ng·mL-1；去乙基奥昔布宁在0.05～25 ng·mL-1线性关系良好(r=0.998 5),最低定量浓度为0.05 ng· 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.

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

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.