Unit of Legal Medicine, Department of Medical and Surgical Sciences, University of Bologna, Via Irnerio 49, Bologna 40126, Italy.
Unit of Legal Medicine, Department of Medical and Surgical Sciences, University of Bologna, Via Irnerio 49, Bologna 40126, Italy.
J Pharm Biomed Anal. 2024 Jun 15;243:116054. doi: 10.1016/j.jpba.2024.116054. Epub 2024 Feb 17.
In the dynamic universe of new psychoactive substances (NPS), the identification of multiple and chemically diverse compounds remains a challenge for forensic laboratories. Since hair analysis represents a gold-standard to assess the prevalence of NPS, which are commonly detected together with classical drugs of abuse (DoA), our study aimed at developing a wide-screen method to detect and quantify 127 NPS and 15 DoA on hair.
A multi-analyte ultra-high performance liquid chromatography mass spectrometry method for the identification and quantification of 127 NPS (phenethylamines, arylcyclohexylamines, synthetic opioids, tryptamines, synthetic cannabinoids, synthetic cathinones, designer benzodiazepines) and 15 DoA in hair samples was developed. A full validation was performed according to the European medicines Agency (EMA) guidelines, by assessing selectivity, linearity, accuracy, precision, limit of quantification (LOQ), limit of detection (LOD), matrix effect and recovery. As a proof of the applicability, the method was applied to 22 authentic hair samples collected for forensic purposes.
Successful validation was achieved, by meeting the required technical parameters, for 137 compounds (122 NPS and 15 DoA), with LOQ set at 4 pg/mg for 129 compounds, at 10 pg/mg for 6 and at 40 pg/mg for 2. The method was not considered validated for 5 NPS, as LLOQ resulted too high for a forensic analysis (80 pg/mg). Among authentic forensic samples, 17 tested positive for DoA, and 10 to NPS, most samples showing positivity for both. Detected NPS were ketamine and norketamine, 5-MMPA, ritalinic acid, methoxyacetyl fentanyl, methylone and RCS-4.
The present methodology represents an easy, low cost, wide-panel method for the quantification of 122 NPS and 15 DoA, for a total of 137 analytes, in hair samples. The method can be profitably applied by forensic laboratories. Similar multi-analyte methods on the hair matrix might be useful in the future to study the prevalence of NPS and the co-occurrence of NPS-DoA abuse.
在新精神活性物质（NPS）的动态世界中，鉴定多种化学结构多样的化合物仍然是法医实验室面临的挑战。由于毛发分析是评估 NPS 流行情况的金标准，而 NPS 通常与传统的滥用药物（DoA）一起被检测到，因此我们的研究旨在开发一种广泛的方法来检测和定量分析毛发中的 127 种 NPS 和 15 种 DoA。
本研究建立了一种多分析物超高效液相色谱-质谱法，用于鉴定和定量分析毛发中的 127 种 NPS（苯乙胺类、芳基环己基胺类、合成阿片类、色胺类、合成大麻素类、合成卡西酮类、苯并二氮䓬类）和 15 种 DoA。根据欧洲药品管理局（EMA）的指南，通过评估选择性、线性、准确性、精密度、定量限（LOQ）、检测限（LOD）、基质效应和回收率，对该方法进行了全面验证。作为适用性的证明，该方法应用于 22 个为法医目的收集的真实毛发样本。
成功验证了 137 种化合物（122 种 NPS 和 15 种 DoA），满足了所需的技术参数，其中 129 种化合物的 LOQ 设定为 4pg/mg，6 种化合物的 LOQ 设定为 10pg/mg，2 种化合物的 LOQ 设定为 40pg/mg。由于 5 种 NPS 的 LLOQ 对于法医分析来说过高，因此未将其视为已验证。在真实的法医样本中，有 17 个样本检测出 DoA 阳性，10 个样本检测出 NPS 阳性，大多数样本同时呈阳性。检测到的 NPS 包括氯胺酮和去甲氯胺酮、5-MMPA、利他林酸、甲氧基乙酰芬太尼、甲基酮和 RCS-4。
本研究建立的方法是一种简单、低成本、广泛的方法，可用于定量分析毛发样本中的 122 种 NPS 和 15 种 DoA，总共 137 种分析物。该方法可被法医实验室应用。未来，类似的毛发基质多分析物方法可能有助于研究 NPS 的流行情况和 NPS-DoA 滥用的共同发生情况。
