University of Camerino, Piazza Cavour 19/f, 62032, Camerino, Italy.
Department of Excellence of Biomedical Sciences and Public Health, University "Politecnica delle Marche" of Ancona, Via Conca 71, 60124, Ancona, Italy.
Anal Bioanal Chem. 2020 Aug;412(21):5125-5145. doi: 10.1007/s00216-020-02462-6. Epub 2020 Feb 15.
Interest on keratinized matrix analysis for clinical and forensic purposes has been recently grown due to the wide temporary detection window for psychotropic and toxic substances entrapped after repeated consumption. The aim of this study was the development and full validation of an UHPLC-MS/MS screening method to quantify 119 molecules among most abused classic drugs and new psychoactive substances in hair and in nails, to assess the polyconsumption. Twenty-five milligrams of hair or nail samples, added with the internal standard mixture, were cut and incubated with 500 μL M3® buffer reagent at controlled temperature. After cooling, 1 μL supernatant was injected in the chromatographic system equipped with an Oasis HLB column. After the 10 min chromatographic separation through a gradient mobile phase (aqueous ammonium formate, phase A; acetonitrile, phase B), the target compounds were detected in multiple reaction monitoring mode. The method was linear (r always better than 0.99) in a calibration range of LOQ 20000 pg compound for milligram hair and of LOQ 1000 pg compound per milligram nail. Process efficiency of analytes under investigation was always better than 65% and no significant ion suppression due to matrix effect was observed. Intra-assay and inter-assay precision and accuracy were always better than 15%. The applicability and trueness of the method were examined by analysing real samples of hair and nail from users of psychoactive drugs in recreational contexts. Both classic drugs and new psychoactive substances could be determined as result of single or repeated use and accumulation in keratin matrices. Graphical abstract.
由于反复摄入后被困的精神药物和有毒物质具有广泛的临时检测窗口，因此最近人们对角蛋白基质分析在临床和法医学方面的兴趣日益浓厚。本研究的目的是开发和全面验证一种 UHPLC-MS/MS 筛选方法，以定量分析头发和指甲中 119 种最常滥用的经典药物和新型精神活性物质，评估多种物质的共同摄入情况。将 25 毫克头发或指甲样本与内标混合物一起切割，并在控温下用 500μL M3®缓冲试剂孵育。冷却后，将 1μL 上清液注入配备 Oasis HLB 柱的色谱系统。在通过梯度流动相（甲酸铵水溶液，A 相；乙腈，B 相）进行 10 分钟色谱分离后，以多反应监测模式检测目标化合物。该方法在毫克头发中每个化合物的定量下限为 20000pg，毫克指甲中每个化合物的定量下限为 1000pg 的校准范围内具有线性（r 始终大于 0.99）。在调查的分析物的过程效率始终高于 65%，并且没有观察到由于基质效应而导致的显著离子抑制。日内和日间精密度和准确度始终优于 15%。通过分析娱乐性使用精神药物的使用者的头发和指甲的实际样本，检验了该方法的适用性和真实性。经典药物和新型精神活性物质都可以作为单次或重复使用以及在角蛋白基质中积累的结果来确定。
