Hepatoprotective effect of tiliroside and characterization of its metabolites in human hepatocytes by ultra-high performance liquid chromatography-high resolution mass spectrometry

Tiliroside, as a dietary flavonoid, has great potential in preventing or treating non-alcoholic fatty liver disease (NAFLD). However, research on tiliroside metabolism remains unclear, limiting its further utilization in healthcare. Herein, we explored the hepatoprotective effect of tiliroside and performed an UHPLC-Q-Exactive Orbitrap/MS-based set of experiments to identify metabolites and metabolic pathways of tiliroside in human hepatocytes. Results showed that tiliroside can exert antioxidant activity to protect against palmitic acid (PA)induced oxidative injury in hepatocytes. Moreover, tiliroside was found to be considerably transformed through metabolism in human hepatocytes based on full MS and MS/MS analysis. Fifteen metabolites derived from the glucuronidation, sulfation, hydrolysis, and oxidation of tiliroside were identified. Glucuronidation and sulfation played key roles in tiliroside metabolism, accounting for approximately 32% and 20% of metabolism after 240 min incubation, respectively. The findings could provide a scientific basis for a more rational and effective application of tiliroside in healthcare.