Synthesis, Detection, and Hepatotoxic Evaluation of Novel Cathinones Recently Identified in the Night-Time Economy

Dawber, Emily Jane (2025) Synthesis, Detection, and Hepatotoxic Evaluation of Novel Cathinones Recently Identified in the Night-Time Economy. Masters by Research thesis (MPhil), Manchester Metropolitan University.

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Abstract

The rapid emergence of novel synthetic cathinones, commonly found in night-time economy settings, presents significant challenges for public health and forensic science. This research focuses on the synthesis of derivatised 3,4-methylenedioxycathinones, detection of the synthesised products, and hepatotoxic evaluation of several newly synthesised synthetic cathinones. Five cathinones; pentylone, dimethylpentylone, N-ethylpentylone (NEP), butylone and putylone, were successfully synthesized with varying yields of 9-65%, using established organic chemistry methods. The synthesised products were fully characterized through 1H-NMR, 13C{1H}-NMR, 1H-1H COSY NMR, HMBC, HSQC, DEPT-135, ATR-FTIR and GC-MS to confirm their structures and purity. A robust GC-MS method was developed and validated for the simultaneous detection of these compounds, demonstrating high sensitivity and specificity. The hepatotoxic potential of the synthesized cathinones was assessed using in vitro cytotoxicity assays on LX2 and HepG2 cell lines. Results indicated varying degrees of cytotoxicity among the compounds. In LX-2 cell lines, MDPV induced a concentration-dependent increase in cytotoxicity, with a significant elevation observed at 1.8 µM. Initial HepG2 assays recorded at 24hrs after dosing, indicated minimal to no cytotoxicity for all compounds at lower concentrations, reflected by minimal cytotoxicity percentage in respects to the maximum cytotoxicity (Obtained by Promega Lysis Buffer). All synthetic cathinones showed similar toxicity profiles between 0.01 and 1 mM, followed by a reduction in cytotoxicity at 2 mM, possibly indicative of cellular adaptation, compound degradation, or assay saturation effects at higher concentrations. MDPHP and putylone exhibited a dose-dependent increase in cytotoxicity, showing a higher cytotoxicity than the other cathinones tested, yet remained below the positive control. These findings suggest that the consumption of these substances may pose possible health risks to users who continuously abuse them or from single high dose usage. This study not only provides critical insights into the toxicological profiles of these emerging synthetic cathinones but also contributes to the development of reliable analytical methods for their detection in forensic contexts. The outcomes underscore the need for ongoing monitoring and regulation of these substances to safeguard public health.