In vitro metabolic profile of mexedrone, a mephedrone analog, studied by high‐ and low‐resolution mass spectrometry

Mexedrone is a synthetic cathinone structurally related to mephedrone, which belongs to the class of N‐alkyl cathinone derivatives, whose metabolic profile has not been fully clarified yet. This study considers the in vitro phase I metabolism of mexedrone, to pre‐select the most appropriate marker(s) of intake. Mexedrone was incubated in the presence of either human liver microsomes or single recombinant CYP450 isoforms. The metabolic profile was outlined by ultra‐high‐performance liquid chromatography coupled to both high‐ and low‐resolution mass spectrometry. In detail, the phase I metabolic profile of mexedrone was initially defined by a time‐of‐flight analyzer, while the chemical structures of the detected metabolites and the potential presence of minor metabolites were subsequently studied by tandem mass spectrometry, using a triple quadrupole analyzer. The main phase I metabolic reactions were hydroxylation and N‐ and O‐dealkylation. The CYP450 isoforms most involved were CYP2C19, responsible for the formation of both hydroxylated and dealkylated metabolites, followed by CYP2D6 and CYP1A2, involved in the hydroxylation reactions only. Finally, a significant fraction of mexedrone unchanged was also detected. Based on this evidence, the most appropriate markers of intake are mexedrone unchanged and the hydroxylated metabolites.