Benzimidazole opioids, often referred to as nitazenes, represent a subgroup of new psychoactive substances with a recent increase in fatal overdoses in the US and Europe. With a variety of analogs emerging on the illicit drug market, forensic labs are challenged to identify these potent drugs. We here present a simple quantitative approach for the determination of nine nitazene analogs, namely, clonitazene, etodesnitazene, etonitazene, etonitazepyne, flunitazene, isotonitazene, metodesnitazene, metonitazene, and protonitazene in whole blood using liquid-phase microextraction and electromembrane extraction in a 96-well format and liquid chromatography tandem mass spectrometry. Green and efficient sample preparation was accomplished by liquid-phase microextraction in a 96-well format and resulted in high extraction yields for all analytes (> 81%). Here, blood diluted with buffer (1:1, %v) was extracted from a donor compartment across a thin organic liquid membrane and into an aqueous acceptor solution. The acceptor solution was collected and directly injected into the analysis platform. Chromatographic separation was accomplished with a biphenyl column, allowing for baseline separation of the structural isomers isotonitazene and protonitazene before detection by multiple reaction monitoring. The validation was performed according to Scientific Working Group of Forensic Toxicology guidelines. The calibration range was from 0.5 to 50 nM (except for protonitazene and clonitazene from 0.1 nM) with good linearity and limits of detection down to 0.01 nM. An AGREEprep assessment was performed to evaluate sample preparation greenness, with a final score of 0.71. Nitazenes represent a current threat to public health, and analytical methods that cover a wide range of these analogs are limited. The here described method may assist in the detection of nitazenes in whole blood and prevent these substances from being missed in post-mortem investigations.

中文翻译：

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通过微萃取和 UHPLC-MS/MS 对新兴“硝氮”苯并咪唑阿片类药物类似物进行高通量定量。

苯并咪唑阿片类药物通常被称为硝氮类药物，是新型精神活性物质的一个亚类，最近在美国和欧洲致命的过量用药有所增加。随着非法药物市场上出现各种类似药物，法医实验室面临着识别这些强效药物的挑战。我们在这里提出了一种简单的定量方法，使用液相微萃取和电膜萃取在 96-孔格式和液相色谱串联质谱法。通过 96 孔形式的液相微萃取实现绿色高效的样品制备，所有分析物的萃取率都很高 (> 81%)。此处，用缓冲液（1:1，%v）稀释的血液通过薄有机液膜从供体室中提取并进入受体水溶液中。收集受体溶液并直接注入分析平台。使用联苯柱完成色谱分离，允许在通过多反应监测检测之前对结构异构体异硝氮酮和质子硝氮进行基线分离。验证是根据法医毒理学科学工作组指南进行的。校准范围为 0.5 至 50 nM（质子氮酮和氯硝氮除外 0.1 nM），线性良好，检测限低至 0.01 nM。进行 AGREEprep 评估来评估样品制备的绿色度，最终得分为 0.71。硝氮烯目前对公共健康构成威胁，而涵盖广泛这些类似物的分析方法是有限的。这里描述的方法可以帮助检测全血中的氮氮烯并防止在尸检调查中遗漏这些物质。