After being incubated with animal and human liver microsomes, metabolites of phase I and II were investigated. A comparison was performed by ultrahigh performance liquid chromatography-quadrupole/time-of-flight coupled to mass spectrometry (UHPLC-Q/TOF). Consequently, a total of four phase I metabolites and three glucuronide binding metabolites of T-2 toxin were discovered. Although a significant metabolic difference was observed among six species, HT-2 toxin was the major product in all species. In addition, the in vivo metabolism of T-2 toxin after oral administration was also investigated in chickens, In total, 18 metabolites were detected, of which 13 were novel, to our knowledge, and reported for the first time. To elucidate the structures of these metabolites, besides accurate mass data from their MS and MS spectra, online hydrogen/deuterium (H/D) exchange technique was also carried out. These new metabolites were regarded as 3'-hydroxy-T-2 3-sulfate, 3'-hydroxy-HT-2 3-sulfate, 4'-hydroxy-HT-2, 3',4'-dihydroxy-HT-2, 4'-carboxyl-T-2, 4'-carboxyl-HT-2, 4'-carboxyl-4'-hydroxy-T-2, and their isomers, implying that T-2 toxin was metabolized more extensively in animals than previously thought. Furthermore, 3'-hydroxy-HT-2, 4'-carboxyl-T-2, 3'-hydroxy-T-2, HT-2 toxin, and neosolaniol were identified to be the major metabolites of T-2 toxin in chickens. The present study expands existing knowledge about T-2 toxin metabolism, informing assessments of the impact T-2 toxin exposure and metabolism on health.
