超高效液相色谱-串联质谱法同时检测人血浆中11种镇静催眠药浓度

Detection and Validated Quantification of 11 Sedative-Hypnotics in Human Plasma by UPLC-MS / MS Method

目的建立超高效液相色谱-串联质谱(UPLC-MS/MS)测定人血浆中11种镇静催眠药的浓度。方法血样采用二氯甲烷∶正己烷∶乙酸乙酯(5∶4∶1)进行萃取,Waters ACQUITY UPLC HSS T3(2.1 mm×100 mm,1.8μm)色谱柱进行分离,乙腈-0.1%氨溶液为流动相进行梯度洗脱,流速0.2 m L·min-1,通过电喷雾离子源(ESI),多重反应监测(MRM)正离子模式进行检测。结果 11种镇静催眠药在所测质量浓度范围内呈现良好的线性关系,相关系数均>0.99,除右佐匹克隆检测限为200 pg·m L-1,其他物质检测限为10~20 pg·m L-1,11种被分析物的日内、日间RSD均<15.0%,提取回收率范围为72.3%~108.0%,基质效应范围为0.86~1.12。结论该方法快速、灵敏、准确,适用于人血浆中11种镇静催眠药浓度的同时检测。

Objective To develop a rapid ultra high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) for determination of 11 sedative-hypnotics in human plasma.Methods The plasma samples were extracted with dichloromethane:n-hexane:acetoacetate(5∶4∶1).The separation was performed on a Waters ACQUITY UPLC HSS T3column(2.1 mm×100 mm,1.8 μm) using the mobile phase composed of acetonitrile-0.1% ammonium solution at a flow rate of0.2 m L·min-1in gradient elution mode.The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring(MRM) via electrospray ionization(ESI) source in positive mode.Results Good linear relation was obtained over the investigated concentration range,with all correlation coefficients higher than 0.99.The limit of detection was 200 pg· m L-1for dexzopiclone,and 10-20 pg · m L-1for other sedative-hypnotics.The intra-and inter-day RSD of 11sedative-hypnotics were no more than 15.0%.The recoveries were in the range of 72.3%-108.0%,and the matrix effects were approximately 0.86-1.12.Conclusion The method is rapid,sensitive and reliable,and suitable for the simultaneous determination of 11 sedative-hypnotics in human plasma.