Unit of Forensic Medicine, Department of Diagnostics and Public Health, University of Verona, Verona, Italy; Institute of Translational Medicine and Biotechnology, Sechenov First Moscow State Medical University, Moscow, Russia.
Unit of Forensic Medicine, Department of Diagnostics and Public Health, University of Verona, Verona, Italy.
J Chromatogr B Analyt Technol Biomed Life Sci. 2020 Sep 1;1152:122263. doi: 10.1016/j.jchromb.2020.122263. Epub 2020 Jul 15.
The use of chromatography hyphenated with mass spectrometry is a well-established approach in clinical and forensic toxicology, particularly for the analysis of the so-called alternative matrices (hair, nails, oral fluid, sweat). This procedure for the quantitative determination of targeted analytes has been reported since the early 1980s and today is the golden standard in analytical toxicology. However this technology has not found wide application the broad spectrum preliminarily screening of samples which is still mostly based on immunoassays. The aim of the present work was to test a recent instrumental approach based on UHPLC-Ion Trap-MS (Toxtyper®, Bruker Daltonics) intended to be used in routine contexts for the analysis of drug of abuse applied to hair toxicological analysis. The reported analytical method is based on a simple hair pre-treatment consisting of an overnight acid incubation in 0.1 mol/L HCl, followed by direct injection, after neutralization with equimolar amount of NaOH. The separation was then performed using a reverse phase column with a rapid gradient elution of 11 min (from 1% acetonitrile in 0.1% ammonium formate to 95% acetonitrile in 0.1% ammonium formate). Detection was by a fast ion trap analyzer (32,500 m/z sec) operating in the mass range 70-800 m/z. The chromatographic retention time and MS2/MS3 data were used for compound identification using a proprietary database which allowed to screen for up to 987 compounds. The tested analytical method showed limits of detection in the range between 0.01 and 0.09 ng/mg of hair matrix for a panel of 16 drugs of abuse (except for MDA, morphine, 6-MAM and norketamine, which showed limits of detection of 0.25, 0.15, 0.15 and 0.25 ng/mg, respectively). The method was validated according to international guidelines on a selected panel of drugs of abuse. The analytical performance of the instrument was assessed by analyzing 968 hair samples from forensic cases. A good concordance with a reference confirmatory method based on GC-MS was found in terms of classification of both negative and positive samples. Finally, the method was also successfully tested by analyzing 12 proficiency test samples containing not only common drugs of abuse but also new psychoactive substances, including fentanyls and cathinones.
色谱-质谱联用在临床和法医毒理学中是一种成熟的方法，特别是用于分析所谓的替代基质（头发、指甲、唾液、汗液）。自 20 世纪 80 年代初以来，人们就一直在报告这种用于定量测定目标分析物的方法，如今它已成为分析毒理学的金标准。然而，这项技术并没有在样品的广谱初步筛选中得到广泛应用，而这种筛选仍然主要基于免疫测定。本工作的目的是测试一种基于超高效液相色谱-离子阱质谱（Toxtyper®，布鲁克·道尔顿公司）的新仪器方法，该方法旨在用于滥用药物的头发毒理学分析的常规分析。所报道的分析方法基于一种简单的头发预处理方法，包括在 0.1 mol/L HCl 中过夜酸孵育，然后直接注射，用等摩尔量的 NaOH 中和。然后使用反相柱进行分离，快速梯度洗脱 11 分钟（从 1%乙腈中的 0.1%甲酸铵到 95%乙腈中的 0.1%甲酸铵）。检测是通过快速离子阱分析仪（32,500 m/z sec）在 70-800 m/z 的质量范围内进行的。色谱保留时间和 MS2/MS3 数据用于使用专有的数据库进行化合物鉴定，该数据库可用于筛选多达 987 种化合物。所测试的分析方法显示，对于一组 16 种滥用药物（除 MDA、吗啡、6-MAM 和去甲氯胺酮外，其检测限分别为 0.01 至 0.09 ng/mg 头发基质），其检测限在 0.01 至 0.09 ng/mg 头发基质范围内。该方法按照国际指南对选定的滥用药物进行了验证。通过分析 968 份法医案件中的头发样本，对仪器的分析性能进行了评估。在对阴性和阳性样本的分类方面，与基于 GC-MS 的参考确证方法具有良好的一致性。最后，该方法还成功地用于分析了 12 个能力验证样本，其中不仅包含常见的滥用药物，还包含新的精神活性物质，包括芬太尼和卡他碱。
