New screening and quantification method by UHPLC-MS/MS of 36 new synthetic cathinones in hair. Application to real cases

Alfredo Fabrizio Lo Faro , Anastasio Tini , Giorgi Kobidze , Giulio Mannocchi , Massimo Gottardi , Francesco Paolo Busardò , Giuseppe Basile , Bezhan Chakvetadze
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Abstract

A class of β-keto analogs of phenethylamine known as synthetic cathinones has been identified as the most emergent new class of psychoactive chemicals in the last ten years. Synthetic cathinones (SC) are becoming more varied, which represents a serious danger to social security and public health worldwide. In this work, an analytical technique based on UHPLC-MS/MS was developed for the simultaneously measurement of 36 synthetic cathinones and two metabolites in hair keratin samples. The separation was conducted by Atlantis Premier BEH C18 AX (2.5 μm, 2.1 × 100 mm) column using elution with 5 mM ammonium acetate in water with 0.1 % formic acid (v/v) (mobile phase A) and 2 mM ammonium formate in MeOH/Acetonitrile 50/50, +0.1 % formic acid (v/v) (mobile phase B). The analysis time was 11 min. Mass spectrometer was equipped with electrospray ionization (ESI). Multiple reaction monitoring (MRM) mode was used for the detection and quantification of the studied compounds. The methodology was successfully validated according to the Organization Scientific Area Committee guidelines, with linearity (r2 ≥ 0.99) from LOQ to 500 pg/mg concentrations for all the compounds investigated. This method was subsequently applied to nine hair samples positive for ten different synthetic cathinones. The most common SCs identified were 3,4-methylenedioxypyrovalerone (3,4-MDPV), in a concentration range 6.0–1000.0 pg/mg, along with alpha-pyrrolidinopentiophenone (α-PHP (54.0 and 554.0 pg/mg, respectively)), followed by the two positional isomers 3-MMC (556.0 and 5000.0 pg/mg) and 4-MMC (11.5 and 448.0 pg/mg). In conclusion, a validated LC-MS/MS method with high specificity was developed offering an easy and affordable sample preparation, and a run time that makes it suitable for use in a high throughput forensic laboratory for the multi-analyte quantification of 36 novel synthetic cathinones and 2 metabolites in hair.

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利用超高效液相色谱-质谱/质谱法筛选和定量毛发中 36 种新型合成卡西酮的新方法。在实际案例中的应用
合成卡西酮(synthetic cathinones)是一类苯乙胺的β-酮类似物,被认为是近十年来最新潮的精神活性化学品。合成卡西酮(SC)的种类越来越多,对全世界的社会安全和公众健康构成了严重威胁。本研究开发了一种基于超高效液相色谱-质谱/质谱的分析技术,用于同时测定头发角蛋白样本中的 36 种合成卡西酮和两种代谢物。采用 Atlantis Premier BEH C18 AX(2.5 μm,2.1 × 100 mm)色谱柱,以 5 mM 乙酸铵水溶液加 0.1 % 甲酸(体积分数)(流动相 A)和 2 mM 甲酸铵水溶液加 MeOH/Acetonitrile 50/50,+0.1 % 甲酸(体积分数)(流动相 B)进行分离。分析时间为 11 分钟。质谱仪配有电喷雾离子化装置(ESI)。多反应监测(MRM)模式用于检测和定量所研究的化合物。该方法成功地通过了组织科学领域委员会准则的验证,从 LOQ 到 500 pg/mg 浓度的线性(r2 ≥ 0.99)适用于所有研究化合物。随后,对 10 种不同合成卡西酮呈阳性的 9 份头发样本采用了该方法。最常见的 SCs 是 3,4-亚甲基二氧吡咯戊酮(3,4-MDPV),浓度范围为 6.0-1000.0 pg/mg,以及α-吡咯烷基苯乙酮(α-PHP,分别为 54.0 和 554.0 pg/mg),其次是两种位置异构体 3-MMC(556.0 和 5000.0 pg/mg)和 4-MMC(11.5 和 448.0 pg/mg)。总之,本研究开发了一种经过验证的 LC-MS/MS 方法,该方法特异性强,样品制备简便且成本低廉,运行时间短,适合在高通量法医实验室中对头发中的 36 种新型合成卡西酮和 2 种代谢物进行多分析物定量分析。
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