超分子溶剂萃取-气相色谱-串联质谱法检测尿液中苯二氮?类药物

目的建立一种尿液中9种苯二氮?类药物的超分子溶剂样品气相色谱-串联质谱(gas chromatography-tandem mass spectrometry,GC-MS/MS)分析方法。方法含9种苯二氮?类药物对照品的尿液样品用四氢呋喃和1-己醇组成的超分子溶剂进行液液萃取,取溶剂层氮吹至干,残余物用甲醇复溶后进行GC-MS/MS分析,数据采集方式为多反应监测模式,采用内标法定量。结果尿液中地西泮、咪达唑仑、氟硝西泮和氯氮平质量浓度在1~100ng/mL,劳拉西泮和阿普唑仑质量浓度在5~100ng/mL,硝西泮和氯硝西泮质量浓度在2~100ng/mL,艾司唑仑在质量浓度0.2~100ng/mL范围内具有良好的线性关系,相关系数为0.9991~0.9999,定量下限为0.2~5ng/mL,提取回收率为81.12%~99.52%,日内精密度相对标准偏差(relative standard deviation,RSD)]和准确度(偏倚)分别小于9.86%、9.51%;日间精密度(RSD)和准确度(偏倚)分别小于8.74%、9.98%。室温和-20℃条件下,尿液中9种药物在15d内具有良好的稳定性。8名志愿者单摄口服阿普唑仑片后,在8~72h内尿液中阿普唑仑的质量浓度为6.54~88.28ng/mL。结论本研究建立的尿液中9种苯二氮?类药物的超分子溶剂萃取-GC-MS/MS分析方法,简便、快速、准确、灵敏,可为临床治疗及司法鉴定中苯二氮?类药物中毒监测提供技术支持。

Supramolecular Solvent Extraction-Gas Chromatography-Tandem Mass Spectrometry for Detection of Benzodiazepines in Urines

Objective To establish a method using supramolecular solvent and gas chromatography-tandem mass spectrometry(GC-MS/MS)to analyze 9 benzodiazepines in urines.Methods Urine samples containing 9 benzodiazepines reference substance were subjected to liquid-liquid extractions with supramolecular solvent,which consisted of tetrahydrofuran and 1-hexanol.The solvent layer was evaporated to dryness by stream of nitrogen.The residue was reconstituted with methanol,and GC-MS/MS analysis was performed on it.The way of data collection was multiple reaction monitoring(MRM)mode;internal standard method was employed for quantification.Results In urine samples,when the range of mass concentration was 1-100 ng/mL for diazepam,midazolam,flunitrazepam and clozapine,5-100 ng/mL for lorazepam and alprazolam,2-100 ng/mL for nitrazepam and clonazepam,and 0.2-100 ng/mL for estazolam,respectively,good linearities were obtained,correlation coefficients were 0.9991-0.9999,the lower limits of the quantifications ranged from 0.2 to 5 ng/mL,the extraction recovery rates were 81.12%-99.52%.The intra-day precisionrelative standard deviation(RSD)]and accuracy(bias)were lower than 9.86% and 9.51%,respectively;the inter-day precision(RSD)and accuracy(bias)were lower than 8.74%and 9.98%,respectively.Nine drugs in urine samples showed good stability at ambient temperature and-20℃ within 15 days.The mass concentrations of alprazolam in urine samples obtained from 8 volunteers who took alprazolam tablets orally within 8-72 h after ingestions ranged from 6.54 to 88.28 ng/mL.Conclusion The supramolecular solvent extraction GC-MS/MS method for analysis of 9 benzodiazepines in urines provided by this study is simple,fast,accurate and sensitive,which can provide technical support for monitoring of poisoning by benzodiazepines for clinical treatment and judicial identification.