Fernández María Del Mar Ramírez, Di Fazio Vincent, Wille Sarah M R, Kummer Natalie, Samyn Nele
Federal Public Service Justice, National Institute of Criminalistics and Criminology, Brussels, Belgium.
Federal Public Service Justice, National Institute of Criminalistics and Criminology, Brussels, Belgium.
J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Aug 15;965:7-18. doi: 10.1016/j.jchromb.2014.05.055. Epub 2014 Jun 6.
Forensic testing for drugs of abuse in hair has become a useful diagnostic tool in determining chronic drug use as well as examining long-term drug history thorough segmental analysis. However, sensitive and specific analytical methods are needed. A simple, rapid and highly sensitive and specific method for the extraction and quantification of 33 opioids, opiates, cocaine, and amphetamines is presented. The method was fully validated according to international guidelines. Twenty milligrams of hair sample was pulverized and then incubated in the same disposable tube with methanol (under sonication at 45°C) during 4h. After centrifugation the supernatant was evaporated up to about 100 μL and a solid phase extraction (SPE) followed by separation and quantification using ultra performance liquid chromatography-tandem mass spectrometry (UHLC-MS/MS) were carried out. Chromatographic separation was achieved using a BEH phenyl column eluted with 0.1% formic acid: methanol (0.1% formic acid). Selectivity of the method was achieved by a combination of retention time, and two precursor-product ion transitions. Good intra-assay and inter-assay precision (relative standard deviations (RSDs) were observed (<15%) for most of the compounds. The lower limit of quantification was fixed at the lowest calibrator in the linearity experiments and it ranged from 0.006 to 0.063 ng/mg. No instability was observed in processed samples. Extraction efficiency varied from 37 to 107% (except for EDDP with a recovery of 5%) and matrix effects ranged from 52 to 160%, and for most of the compounds it was compensated by the internal standard (IS). The method was subsequently applied to authentic hair samples obtained from forensic and toxicology cases and to proficiency test (obtaining z-scores lower than 1 for most of the compounds). The validation and actual sample analysis results show that this method is rugged, precise, accurate, and well-suited for routine hair analysis.
毛发中滥用药物的法医检测已成为确定长期药物使用情况以及通过分段分析全面检查长期用药史的一种有用诊断工具。然而，需要灵敏且特异的分析方法。本文介绍了一种简单、快速、高度灵敏且特异的方法，用于提取和定量33种阿片类药物、鸦片制剂、可卡因和苯丙胺类药物。该方法已按照国际指南进行了全面验证。取20毫克毛发样本研磨后，在同一一次性试管中与甲醇一起（于45°C超声处理）孵育4小时。离心后，将上清液蒸发至约100微升，接着进行固相萃取（SPE），然后使用超高效液相色谱 - 串联质谱法（UHLC - MS/MS）进行分离和定量。使用以0.1%甲酸：甲醇（0.1%甲酸）洗脱的BEH苯基柱实现色谱分离。通过保留时间以及两个前体 - 产物离子跃迁的组合实现该方法的选择性。大多数化合物的批内和批间精密度良好（相对标准偏差（RSD）<15%）。在线性实验中，定量下限固定为最低校准物，范围为0.006至0.063纳克/毫克。在处理后的样本中未观察到不稳定性。提取效率在37%至107%之间（除EDDP回收率为5%外），基质效应在52%至160%之间，并且对于大多数化合物，其通过内标（IS）得到补偿。该方法随后应用于从法医和毒理学案例中获取的真实毛发样本以及能力验证测试（大多数化合物的z分数低于1）。验证和实际样本分析结果表明，该方法耐用、精密、准确，非常适合常规毛发分析。
