In vitro characterization of potential CYP‐ and UGT‐derived metabolites of the psychoactive drug 25B‐NBOMe using LC‐high resolution MS

Abstract

One of the main challenges posed by the emergence of new psychoactive substances is their identification in human biological samples. Trying to detect the parent drug could lead to false-negative results when the delay between consumption and sampling has been too long. The identification of their metabolites could then improve their detection window in biological matrices. Oxidative metabolism by cytochromes P450 and glucuronidation are two major detoxification pathways in humans. In order to characterize possible CYP- and UGT-dependent metabolites of the 2-(4-bromo-2,5-dimethoxy-phenyl)-N-[(2-methoxyphenyl)methyl]ethanamine (25B-NBOMe), a synthetic psychoactive drug, analyses of human liver microsome (HLM) incubates were performed using an ultra-high performance liquid chromatography system coupled with a quadrupole-time of flight mass spectrometry detector (UHPLC-Q-TOF/MS). On-line analyses were performed using a Waters OASIS HLB column (30 x 2.1 mm, 20 µm) for the automatic sample loading and a Waters ACQUITY HSS C18 column (150 x 2 mm, 1.8 µm) for the chromatographic separation. Twenty-one metabolites, consisting of 12 CYP-derived and 9 UGT-derived metabolites, were identified. O-Desmethyl metabolites were the most abundant compounds after the phase I process, which appears to be in accordance with data from previously published NBOMe-intoxication case reports. Although other important metabolic transformations, such as sulfation, acetylation, methylation or glutathione conjugation, were not studied and artefactual metabolites might have been produced during the HLM incubation process, the record of all the metabolite MS spectra in our library should enable us to characterize relevant metabolites of 25B-NBOMe and allow us to detect 25B-MBOMe users.

Detection of the designer benzodiazepine flunitrazolam in urine and preliminary data on its metabolism

“…In vitro HLMs incubations and LC–HRMS analyses were performed in order to investigate the appearance of CYP‐ and UGT‐dependent FNTZ metabolites . Using a software algorithm, four potential FNTZ metabolites were predicted: 4‐hydroxy‐FNTZ, desnitro‐FNTZ, 7‐amino‐FNTZ and 7‐acetamido‐FNTZ which is in agreement with observed in urine from subjects undergoing flunitrazepam treatment .…”

Characterization of hallucinogenic phenethylamines using high‐resolution mass spectrometry for non‐targeted screening purposes

“…Similar to the present study, they postulated that these product ions formed from the loss of NH 3 , •CH 6 N and CH 9 N However, they did not suggest the location where the methyl radicals were lost (product ions 3 and 4) . A study conducted by Boumrah et al . identified that 2C–B was a metabolic product of 25B–NBOMe when analyzed by HRMS.…”

Prevalence of new psychoactive substances and prescription drugs in the Belgian driving under the influence of drugs population

“…Additional compounds such as NPS and medication were detected via two different qualitative methods; one using high resolution mass spectrometry (LC‐HRMS) Xevo‐G2‐QTOF‐XS system (Waters, Manchester, UK) (M1) and the other one an LC–MS/MS Xevo TQS system (Waters, Manchester, UK) (M2). These two already published methods complement one another in specificity (high resolution mass spectrometry of M1) and sensibility (MRM of M2). Briefly, for M1, 400 μL of sulfosalicylic acid (3%) and 100 μL of methyl‐clonazepam and β‐OH‐ethyltheophyllin as internal standards IS were added to 100 μL of plasma.…”