First insight into human extrahepatic metabolism of flame retardants: Biotransformation of EH-TBB and Firemaster-550 components by human skin subcellular fractions.

2-ethylhexyl-2,3,4,5-tetrabromobenzoate (EH-TBB) and a mixture of EH-TBB, Bis(2-ethylhexyl)tetrabromphthalate (BEH-TEBP) and Triphenyl phosphate (TPhP), prepared in a ratio similar to the Firemaster-550™ (FM550) flame retardant formulation, were exposed to human skin subcellular fractions (S9) to evaluate their dermal in vitro metabolism for the first time. After 60 min of incubation, tetrabromobenzoic acid (TBBA) and diphenyl phosphate (DPhP) were identified as the major metabolites of EH-TBB and TPhP, respectively using UPLC-Q-Exactive Orbitrap™-MS analysis. Dermal biotransformation of EH-TBB and TPhP was catalyzed by skin carboxylesterases rather than CYP450 enzymes, while no stable metabolites could be identified for BEH-TEBP. Metabolite formation rates of EH-TBB as individual compound and as a component of FM550 fitted the Michaelis-Menten model, while no steady state could be reached for TPhP under experimental conditions. Estimated maximum metabolic rate (V) for TBBA formation upon exposure to FM550 was lower than V for EH-TBB (1.08 and 15.2 pmol min mg protein, respectively). This indicates dermal metabolism would contribute less to the clearance of EH-TBB body burden than hepatic metabolism (V = 644 pmol min mg protein). Implications for human exposure include EH-TBB accumulation in skin tissue and human exposure to dermal metabolic products, which may have different toxicokinetic and toxicodynamic parameters than parent flame retardants.