Integrated UHPLC-MS/MS and HRMS Strategy for Screening Potentially Hepatotoxic Furan-Containing Compounds in Dioscorea bulbifera L.

Dioscorea bulbifera L. (DBL), a traditional herbal medicine used to treat thyroid disorders and tumors, has raised considerable safety concerns due to its potential hepatotoxic effects. Previous research suggests that this liver injury may be associated with the metabolic activation of furan-containing compounds (FCCs) present in DBL. Here, we systematically characterized FCCs and their reactive metabolites in DBL extract through an integrated analytical approach combining ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) and ultra-high performance liquid chromatography coupled with tandem high-resolution mass spectrometry (UHPLC-HRMS). Employing both targeted and untargeted mass spectrometric analyses, we identified a total of 849 constituents in the aqueous extract of DBL, including 10 furanoditerpenoids and 17 additional FCCs. To elucidate the metabolic activation pathways, we used glutathione (GSH) and 4-bromobenzylamine (BBA) as dual trapping agents within a mouse liver microsomal (MLM) incubation system. Through P450-mediated metabolism, seven corresponding adducts derived from diosbulbin B (DSB), C (DSC), D (DSD), E (DSE), L (DSL), N (DSN), and 2-pentylfuran were successfully trapped and characterized. This study establishes a sensitive and specific approach for the comprehensive profiling of potentially hepatotoxic furan compounds in DBL, and provides valuable insights into the formation of reactive metabolites and toxicity mechanisms related to furan-containing herbal medicine.