Marshall University, College of Science, Forensic Science Graduate Program, 1401 Forensic Science Drive, Huntington, WV 25701, USA.
Palm Beach County Sheriff's Office, 3228 Gun Club Road, West Palm Beach, FL 33406, USA.
J Anal Toxicol. 2021 May 14;45(5):462-474. doi: 10.1093/jat/bkaa109.
Benzodiazepines are a commonly prescribed class of drugs that have the potential for abuse. The Palm Beach County Sheriff's Office received drug seizure submissions that included novel and/or nonroutine benzodiazepines of increasing prevalence from 2017 to 2019. This prompted the development of a method of analysis for these compounds in biological specimens. The method tests for 16 novel and nonroutine benzodiazepines and suvorexant in whole blood by liquid chromatography-tandem mass spectrometry (LC-MS-MS). The target analytes included bromazepam, clobazam, clonazolam, clotiazepam, diclazepam, estazolam, etizolam, flualprazolam, flubromazepam, flubromazolam, loprazolam, lormetazepam, phenazepam, prazepam, suvorexant, tetrazepam and triazolam. The method uses 200 µL of sample, protein precipitation and an instrument run-time of 8 min. The limit of detection was either 1 or 5 ng/mL and the limit of quantitation was either 5 or 25 ng/mL depending on the analyte. The method was validated for quantitative analysis for 15 out of the 17 analytes. Flubromazepam and prazepam were validated for qualitative identification only. A quadratic calibration model (r2 > 0.990) with 1/x weighting was used for all analytes for quantitative analysis. The calibration range was either 5-100 or 25-500 ng/mL depending on the analyte. The coefficient of variation of replicate analyses was within 14% and bias was within ±14%. The method provides a sensitive, efficient and robust procedure for the quantitation and/or qualitative identification of select novel and nonroutine benzodiazepines and suvorexant using LC-MS-MS and a sample volume of 200 µL.
苯二氮䓬类药物是一类常用的处方药物，具有滥用的潜力。从 2017 年到 2019 年，棕榈滩县治安官办公室收到了包括新型和/或非常规苯二氮䓬类药物的药物扣押报告，这些药物的流行率在不断增加。这促使我们开发了一种在生物样本中分析这些化合物的方法。该方法通过液相色谱-串联质谱法（LC-MS-MS）测试全血中的 16 种新型和非常规苯二氮䓬类药物和苏沃雷生。目标分析物包括溴西泮、氯巴占、氯氮䓬、氯硝西泮、地西泮、依替唑仑、艾司唑仑、氟拉䓬仑、氟布仑唑仑、氟马西泮、劳拉西泮、咪达唑仑、苯甲二氮䓬、普拉西泮、苏沃雷生、佐匹克隆和三唑仑。该方法使用 200 µL 样本、蛋白质沉淀和 8 分钟的仪器运行时间。检测限为 1 或 5ng/mL，定量限为 5 或 25ng/mL，具体取决于分析物。该方法对 17 种分析物中的 15 种进行了定量分析验证。氟马西泮和普拉西泮仅进行了定性鉴定验证。所有分析物的定量分析均采用二次校准模型（r2 > 0.990），并使用 1/x 加权。校准范围取决于分析物，范围为 5-100 或 25-500ng/mL。重复分析的变异系数在 14%以内，偏差在±14%以内。该方法提供了一种灵敏、高效和可靠的 LC-MS-MS 程序，可用于定量和/或定性分析选择的新型和非常规苯二氮䓬类药物和苏沃雷生，使用的样本量为 200µL。
