UPLC-MS/MS同时检测人尿液中8种苯二氮䓬类及3种非苯二氮䓬类镇静催眠药

Simultaneous Determination of 8 Kinds of Benzodiazepines and 3 Kinds of Non-benzodiazepines in Human Urine by UPLC-MS/MS

关键词:  超高效液相色谱-串联质谱  镇静催眠药  同时测定  人尿液

Key words:  UPLC-MS/MS  sedativehypnotics  simultaneous determination  human urine

摘要:

目的 建立超高效液相色谱-串联质谱法测定人尿液中8种苯二氮?类及3种非苯二氮?类药物的浓度。方法 尿液样品采用二氯甲烷∶正己烷∶乙酸乙酯(5∶4∶1)进行萃取，Waters ACQUITY UPLC HSS T3色谱柱(2.1 mm×100 mm，1.8 μm)进行分离，乙腈-0.1%氨水溶液为流动相进行梯度洗脱，流速为0.2 mL·min^-1，通过电喷雾离子源，多重反应监测正离子模式进行检测。结果 11种被分析物在所测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0.5 ng·mL^-1，其他物质检测限为0.01~0.02 ng·mL^-1，11种被分析物的日内、日间RSD均<15.0%(n=5)，提取回收率为68.8%~115.0%，基质效应为0.85~1.14。结论 该方法快速、灵敏、准确，适用于同时检测人尿中8种苯二氮?类及3种非苯二氮?类药物浓度。

Abstract:

OBJECTIVE To develop a rapid UPLC-MS/MS method for the determination of 8 kinds of benzodiazepines and 3 kinds of non-benzodiazepines in human urine. METHODS The urine samples were extracted with dichloromethane∶ n-hexane∶acetoacetate(5∶4∶1). The separation was performed on a Waters ACQUITY UPLC HSS T3 column(2.1 mm×100 mm, 1.8 μm) using the mobile phase of acetonitrile-0.1% ammonium solution at a flow rate of 0.2 mL·min^-1 in gradient elution mode. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring via electrospray ionization source in positive mode. RESULTS Good linear relation was obtained over the investigated concentration range, with all correlation coefficients >0.99. The limit detection was 0.5 ng·mL^-1 for dexzopiclone and 0.01-0.02 ng·mL^-1 for other sedativehypnotics. The intra- and inter-day RSDs were <15%(n=5). The recoveries were in 68.8%-115.0%. The matrix effects were approximately 0.85-1.14. CONCLUSION This method is rapid, sensitive, reliable, and suitable for the simultaneous determination of 8 kinds of benzodiazepines, zolpidam, zaleplone and dexzopiclone in human urine.