近年来,涵盖大多数滥用药物类别的数百种新型精神活性物质(NPS)已被引入娱乐性药物市场。最近越来越常见的一类NPS药物是“设计师”苯二氮卓类药物。由于与处方苯二氮卓类药物结构相似,这些物质中的一些可在免疫测定筛选中引起阳性反应(即交叉反应)。因此,在方法确认过程中加入NPS苯二氮卓类药物越来越重要,以确保准确鉴定密切相关的化合物以及检测苯二氮卓类药物本身。这里,我们努力开发一种使用反相液相色谱分离结合高分辨率质谱(LC-HRMS)检测尿液中28种NPS苯二氮卓类药物的筛选和确认方法。MS在ThermoFischerScientificQExactiveOrbitrap仪器上以正电喷雾模式进行,使用全扫描(用于筛选)或平行反应监测(用于确认)。我们发现该方法的定量下限为5至50ng/mL。除了一种分析物之外,所有分析物的筛选和确认的分析精度和准确度均≤15%。该方法用于分析常规药物测试中的患者尿液样本以及急诊病房中出现的急性中毒病例的样本。总之,28种苯二氮卓类药物中的16种(即,Clobazam,氯硝唑仑,去氯替唑,Diclazepam,艾司唑仑,etizolam,氟溴西泮,氟溴唑兰,氟尼唑仑,3-羟基氟溴马西泮,3-羟基苯西泮,氯唑仑,Meclonazepam,美替托兰,nifoxipam,和吡唑仑)在尿液样本中检测到。患者样本分析的结果表明,使用NPS苯二氮卓类药物的患病率很高,强调在药物测试菜单中包括新的滥用药物的重要性。
Hundreds of new psychoactive substances (NPS) covering most drugs-of-abuse classes have been introduced to the recreational drug market in recent years. One class of NPS drugs that has become more common recently is \"designer\" benzodiazepines. Due to a close structural resemblance with prescription benzodiazepines, some of these substances may elicit a positive response (i.e. cross react) in immunoassay screening. Consequently, it is increasingly important to include NPS benzodiazepines during method confirmation to ensure accurate identification of closely-related compounds as well as detection of the benzodiazepines themselves. Here, we present our efforts to develop a screening and confirmation method for detection of 28 NPS benzodiazepines in urine using reversed-phase liquid chromatographic separation in combination with high-resolution mass spectrometry (LC-HRMS). MS was performed in positive electrospray mode on a Thermo Fischer Scientific Q Exactive Orbitrap instrument using either full scan (for screening) or parallel reaction monitoring (for confirmation). We found the lower quantification limit of the method to range from 5 to 50 ng/mL. Analytical precision and accuracy were ≤15% for both screening and confirmation for all except one analyte. The method was used to analyze patient urine samples from routine drug testing and samples from acute intoxication cases presenting in emergency wards. Altogether, 16 of the 28 benzodiazepines (i.e., clobazam, clonazolam, deschloroetizolam, diclazepam, estazolam, etizolam, flubromazepam, flubromazolam, flunitrazolam, 3-hydroxyflubromazepam, 3-hydroxyphenazepam, ketazolam, meclonazepam, metizolam, nifoxipam, and pyrazolam) were detected in the urine samples. The results from patient sample analysis indicate a high prevalence of NPS benzodiazepine use, emphasizing the importance of including novel drugs of abuse in drug testing menus.