Biotransformation of the fungal neurotoxin Thomitrem A by primary rat hepatocytes.

The tremorgenic mycotoxin Thomitrem A is a secondary metabolite produced mainly by the fungus Penicillium crustosum that is frequently found on spoiled stored food and feed. Typical signs of intoxication observed in dogs after the consumption of food waste are emesis, tremors, seizures progressing to ataxia and lack of coordinated movements. How uptake of Thomitrem A relates to exposure is unknown so far since data on biotransformation and toxicokinetics are missing. In this study the toxin was therefore metabolised in an exploratory in vitro experiment by rat hepatocytes, and substrate depletion as well as the formation of hepatic metabolites were investigated. Seven metabolites were characterised by their retention times and fragmentation patterns in LC-MS/MS analysis. They were found to be products of oxidation and dehydration processes and occurred at different incubation time points, showing different signal abundance-time curve profiles. Toxicokinetic parameters were derived from the Thomitrem A depletion curve applying principles of in vitro-to-in vivo extrapolation (IVIVE). The predicted medium maximum bioavailability in rats could be of importance for the assessment of exposure in cases of intoxication if it was confirmed in vivo and in other species.