The metabolism of the psychoactive compound 2C-B-Fly-NBOMe was investigated using three systems: human liver microsomes, the fungus Cunninghamella elegans, and live rats. Thirty-five phase I and nine phase II metabolites were identified. Major metabolic pathways include hydroxylation, O-demethylation, oxidative debromination, and N-demethoxybenzylation, followed by glucuronidation or N-acetylation. Human liver microsomes produced the most metabolites at highest concentrations. Two poly-hydroxylated metabolites appeared only in rat urine, while the fungus generated dehydrogenated, N-oxygenated, and dibrominated metabolites. These findings clarify how the body processes this substance, aiding understanding of its effects and potential toxicity.
Two new immunoassays, a lateral flow immunoassay (LFIA) and an enzyme-linked immunosorbent assay (ELISA), were developed to detect the hallucinogenic phenethylamine 2C-B and related drugs in urine. The LFIA provides rapid on-site screening with a detection limit of 15 ± 7 ng mL⁻¹, suitable for identifying recent exposure. The ELISA offers much higher sensitivity, with a detection limit of 6 ± 3 pg mL⁻¹, and its results closely match those from established UHPLC-MS-MS analysis, making it reliable for quantitative confirmation in toxicology. Both methods address the need for simple, cost-effective tools to monitor these dangerous substances.