液相色谱-串联质谱法同时测定人血浆中奥美拉唑及其代谢物奥美拉唑砜的质量浓度

Simultaneous determination of omeprazole and its metabolite omeprazole sulfone in human plasma by LC-MS/MS

目的建立人血浆中奥美拉唑及其代谢物奥美拉唑砜质量浓度的液相色谱-串联质谱(LC-MS/MS)测定方法,并应用于临床试验中奥美拉唑及其代谢物奥美拉唑砜血药质量浓度的测定。方法以奥美拉唑d3为内标,人血浆样品经蛋白沉淀处理后进行LC-MS/MS分析。采用Luna C18柱(50 mm×2.0 mm,3mm)色谱柱,流动相为甲醇(体积分数0.1%甲酸)-5mmol·L-1醋酸铵缓冲溶液(体积分数0.1%甲酸),使用APCI离子源,正离子多反应检测(MRM)模式检测,用于定量分析的奥美拉唑、奥美拉唑砜和内标奥美拉唑d3的检测离子对分别为m/z:345.9→197.9;m/z:362.2→150.2;m/z:349.1→198.1。药代动力学参数采用(drug and statistics ver 2.0,DAS)统计软件计算。结果测定血浆中奥美拉唑/奥美拉唑砜线性范围为1.56~1600μg·L-1。奥美拉唑及奥美拉唑砜在其标准曲线范围内线性关系良好(r均大于0.99),批内精密度(RSD)及批间精密度(RSD)均不大于11.5%,提取回收率稳定。结论本方法适用于奥美拉唑及其代谢物奥美拉唑砜的血药浓度测定。

Objective The study aims to establish an LC-MS/MS method for the simultaneous determination of omeprazole and its major metabolite omeprazole sulfone in human plasma.Methods Omeprazoled3 was used as the internal standard,The plasma samples were prepared by protein precipitation method,and then separated on a Phenomenex C18(50 mm×2.0 mm,3 mm)column.The mobile phase consisted of methyl alcohol-5 mM acetamide buffer(including 0.1%formic acid).An APCI source was applied and operated in the positive multiple reaction monitoring(MRM)mode,The mass spectrometers were operated in m/z:345.9→197.9 for omeprazole,m/z:362.2→150.2 for omeprazole sulfone,m/z:349.1→198.1 for omeprazoled3.The pharmacokinetics parameters were calculated by DAS 2.0 program.Results The method was validated in concentration ranges of 1.56-1600μg·L-1 for omeprazole/omeprazole sulfone in human plasma,the two compounds showed a good linear relation-ship in the range of line(r>0.99).The intra-day and inter-day precision(RSD)were less than 11.5%.The average extraction recoveries of two compounds were stabilization.Conclusion The method is simple,rapid,sensitive,and accurate for determination of omeprazole and omeprazole sulfone in human plasma and was successfully applied to a human pharmacokinetic study.