Abstract：
Protonitazene is a synthetic benzoimidazole opioid of the nitazenes class, developed in the 1950s as an effective analgesic, but never released on the market due to severe side effects and possible dependence. Despite its increasing use as a new psychoactive substance starting in 2019, its detection in human hair of intoxicated and deceased consumers has never been reported. We present the development and validation of a specific procedure to identify protonitazene in hair by LC-MS-MS. Drugs were incubated overnight at 40°C in 1 mL borate buffer, pH 9.5 with 20 mg pulverized hair and 1 ng/mg fentanyl-d5 used as internal standard. Drugs were then extracted with a mixture of organic solvents. The chromatographic separation was performed using a HSS C18 column with a 15 min gradient elution. Linearity was verified from 1 to 100 pg/mg. The limit of detection was estimated at 0.1 pg/mg. No interference was noted from a large panel of natural and synthetic opioids, fentanyl derivatives or other new synthetic opioids. Protonitazene was identified at 70 and at > 7600 pg/mg in the whole head hair specimens of two male subjects deceased from acute drug overdose in jail. Protonitazene was also identified at 14 and 54 pg/mg in two living co-prisoners. As nitazenes represent a growing threat to public health in various parts of the world, this method was developed in response to the challenges posed by the identification of this class of substances.
摘要：
Protonitazene是硝基苯类的合成苯并咪唑阿片类药物，在20世纪50年代发展成为一种有效的镇痛药，但由于严重的副作用和可能的依赖性，从未在市场上发布。尽管从2019年开始越来越多地使用它作为一种新的精神活性物质，但从未报道过在醉酒和已故消费者的头发中检测到它。我们提出了通过LC-MS-MS鉴定头发中质子氮烯的特定程序的开发和验证。药物在40℃下在1mL硼酸盐缓冲液中孵育过夜，pH9.5,其中20mg粉碎的毛发和lng/mg芬太尼-d5用作内标。然后用有机溶剂的混合物提取药物。使用具有15min梯度洗脱的HSSC18柱进行色谱分离。从1至100μg/mg验证线性。检测极限估计为0.1pg/mg。没有发现大量天然和合成阿片类药物的干扰，芬太尼衍生物或其他新的合成阿片类药物。在两名因监狱中急性药物过量而死亡的男性受试者的整个头毛标本中，丙氮嗪的含量为70和>7600pg/mg。在两名在世的共同囚犯中还发现了14和54pg/mg的丙尼氮嗪。由于硝基苯在世界各地对公共卫生的威胁越来越大，这种方法是为了应对这类物质鉴定带来的挑战而开发的。
