Section of Forensic Chemistry, Department of Forensic Medicine, Faculty of Health Sciences, University of Copenhagen, 3. DK-2100, Denmark.
J Anal Toxicol. 2010 Jul-Aug;34(6):332-41. doi: 10.1093/jat/34.6.332.
An ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS-MS) method for detection of 23 benzodiazepines and related compounds in whole blood was developed and validated. The method is used for screening and quantitation of benzodiazepines in whole blood received from autopsy cases and living persons. The detected compounds were alprazolam, bromazepam, brotizolam, chlordiazepoxide, demoxepam, clobazam, clonazepam, 7-aminoclonazepam, diazepam, nordiazepam, estazolam, flunitrazepam, 7-aminoflunitrazepam, lorazepam, lormetazepam, midazolam, nitrazepam, 7-aminonitrazepam, oxazepam, temazepam, triazolam, zaleplon, and zopiclone. Whole blood from drug-free volunteers was used for all experiments. Blood samples (0.200 g) were extracted with ethyl acetate at pH 9. Target drugs were quantified using a Waters ACQUITY UPLC system coupled to a Waters Quattro Premier XE triple quadrupole in positive electrospray ionization, multiple reaction monitoring mode. The use of deuterated internal standards for most compounds verified that the accuracy of the method was not influenced by matrix effects. Extraction recoveries were 73-108% for all analytes. Lower limits of quantification ranged from 0.002 to 0.005 mg/kg. Long-term imprecision (CV%) ranged from 6.0 to 18.7%. We present a fully validated UPLC-MS-MS method for 23 benzodiazepines in whole blood with a run-time of only 5 min and using only 0.200 g of whole blood.
建立并验证了一种超高效液相色谱-串联质谱法（UPLC-MS-MS），用于检测全血中的 23 种苯并二氮䓬类药物及相关化合物。该方法用于筛查和定量来自尸检和活体的全血中的苯并二氮䓬类药物。检测到的化合物包括阿普唑仑、溴西泮、布他唑仑、氯氮䓬、地西泮、氯硝西泮、氯巴占、氯氮䓬、7-氨基氯硝西泮、地西泮、去甲西泮、依替唑仑、氟硝西泮、7-氨基氟硝西泮、劳拉西泮、劳拉西泮、咪达唑仑、硝西泮、7-氨基硝西泮、奥沙西泮、替马西泮、三唑仑、扎来普隆和佐匹克隆。所有实验均使用无药物志愿者的全血。在 pH 9 下，用乙酸乙酯提取 0.200 g 全血。采用 Waters ACQUITY UPLC 系统结合 Waters Quattro Premier XE 三重四极杆在正电喷雾电离、多反应监测模式下定量测定目标药物。大多数化合物均使用氘代内标，证明方法的准确性不受基质效应的影响。所有分析物的提取回收率为 73-108%。定量下限范围为 0.002-0.005 mg/kg。长期精密度（CV%）范围为 6.0-18.7%。我们提出了一种完全验证的 UPLC-MS-MS 方法，用于检测全血中的 23 种苯并二氮䓬类药物，运行时间仅为 5 分钟，仅需 0.200 g 全血。
