UPLC-MS/MS快速检测人血浆中7种苯二氮■类药物及其临床应用

Fast determination of 7 benzodiazepines in human plasma by UPLC-MS/MS and its clinical application

目的:建立并验证一种定性定量检测人血浆中7种常见苯二氮[艹卓]类药物的方法,并将其应用于临床检测。方法:血浆样品经乙腈沉淀蛋白后,采用液相-串联质谱联用仪测定。色谱柱为BEH C18,以含0.01%甲酸乙腈溶液-0.01%甲酸水溶液为流动相梯度洗脱,流速为0.4 mL·min^-1,电喷雾正离子模式,多反应监测。结果:劳拉西泮、硝西泮、阿普唑仑、咪达唑仑、三唑仑、氯硝西泮、地西泮浓度分别在62.5~2 000(r=0.999 8)、62.5~2 000(r=0.999 7)、31.25~1 000(r=0.999 9)、62.5~2 000(r=0.999 7)、6.25~200(r=1)、62.5~2 000(r=0.999 7)、625~20 000(r=0.996 8)ng·mL^-1范围内线性关系良好;定量下限分别为7.8,3.9,1.9,3.9,3.1,3.9,19.5 ng·mL^-1;日内和日间精密度RSD均小于14.75%,方法回收率均在87.84%~116.06%之间;提取回收率均在74.55%~96.72%之间;基质效应均在81.68%~97.20%之间;在多种储存条件下,处理前后的血浆样品稳定性好。结论:本方法检测速度快、专属性高、重复性好、定性能力强,可有效检测治疗及中毒浓度范围内的7种苯二氮[艹卓]类药物。

OBJECTIVE To establish and validate a qualitative and quantitative method for the determination of 7 common benzodiazepines in human plasma, which can be applied to clinical examination. METHODS Plasma samples were precipitated with acetonitrile and determined by liquid chromatography-tandem mass spectrometry. The column was BEH C18 and eluted with a gradient of 0.01% formic acid in acetonitrile-0.01% formic acid in water as the mobile phase at a flow rate of 0.4 mL·min-1 in ESI positive ion mode with multiple reaction monitoring. RESULTS The linear ranges of lorazepam, niazepam, alprazolam, midazolam, triazolam, clonazepam and diazepam were 62.5-2 000(r=0.999 8), 62.5-2 000(r=0.999 7), 31.25-1 000(r=0.999 9), 62.5-2 000(r=0.999 7), 6.25-200(r=1), 62.5-2 000(r=0.999 7), 625-20 000(r=0.996 8)ng·mL-1, respectively. The lower limit of quantitation were 7.8, 3.9, 1.9, 3.9, 3.1, 3.9, and 19.5 ng·mL-1, respectively. The RSDs of inter-day and intra-day precision were both less than 14.75%. The relative recovery rates were all within the range of 87.84%-116.06%. The extraction recovery rates were within the range of 74.55%-96.72%. The matrix effects were within the range of 81.68%-97.20%. Plasma samples remained stable before and after treatment under various storage conditions. CONCLUSION This method is rapid, specific, accurate and repeatable, which has been successfully used for clinical detection of 7 drugs at different concentrations.