尿液、血清、血浆和胎粪中13种苯二氮䓬类药物及其代谢物的液相色谱-串联质谱分析

LC-MS/MS analysis of 13 benzodiazepines and metabolites in urine, serum, plasma, and meconium.

作者信息

Marin Stephanie J, McMillin Gwendolyn A

机构信息

ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT, USA.

出版信息

Methods Mol Biol. 2010;603:89-105. doi: 10.1007/978-1-60761-459-3_9.

DOI:10.1007/978-1-60761-459-3_9

PMID:20077062

Abstract

We describe a single method for the detection and quantitation of 13 commonly prescribed benzodiazepines and metabolites: alpha-hydroxyalprazolam, alpha-hydroxyethylflurazepam, alpha-hydroxytriazolam, alprazolam, desalkylflurazepam, diazepam, lorazepam, midazolam, nordiazepam, oxazepam, temazepam, clonazepam and 7-aminoclonazepam in urine, serum, plasma, and meconium. The urine and meconium specimens undergo enzyme hydrolysis to convert the compounds of interest to their free form. All specimens are prepared for analysis using solid-phase extraction (SPE), analyzed using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS), and quantified using a three-point calibration curve. Deuterated analogs of all 13 analytes are included as internal standards. The instrument is operated in multiple reaction-monitoring (MRM) mode with an electrospray ionization (ESI) source in positive ionization mode. Urine and meconium specimens have matrix-matched calibrators and controls. The serum and plasma specimens are quantified using the urine calibrators but employing plasma-based controls. Oxazepam glucuronide is used as a hydrolysis control.

摘要

我们描述了一种用于检测和定量尿液、血清、血浆和胎粪中13种常用苯二氮䓬类药物及其代谢物的单一方法：α-羟基阿普唑仑、α-羟基乙基氟西泮、α-羟基三唑仑、阿普唑仑、去烷基氟西泮、地西泮、劳拉西泮、咪达唑仑、去甲地西泮、奥沙西泮、替马西泮、氯硝西泮和7-氨基氯硝西泮。尿液和胎粪样本进行酶水解，将目标化合物转化为游离形式。所有样本均采用固相萃取（SPE）进行分析前处理，采用液相色谱-串联质谱（LC-MS/MS）进行分析，并使用三点校准曲线进行定量。所有13种分析物的氘代类似物作为内标。仪器在多反应监测（MRM）模式下运行，采用电喷雾电离（ESI）源，处于正离子模式。尿液和胎粪样本有基质匹配的校准物和对照。血清和血浆样本使用尿液校准物进行定量，但采用基于血浆的对照。奥沙西泮葡萄糖醛酸苷用作水解对照。