Abstract: OBJECTIVE To investigate the metabolites of flavonoid vitexin in rats, and to identify its metabolic pathways. METHODS The SD rats were intragastric administration vitexin with a dose of 5 mg·kg^-1. The urine were collected at 0-3 h, 3-6 h, 6-12 h after drug administration. UPLC-Q-TOF was applied to separate and analyze vitexin and its metabolites in rat urine. RESULTS A total of 3 vitexin metabolites were detected and identified by Metabolynx XS software, which structural identification was based on mass fragment information. CONCLUSION The results show that vitexin is detected in rat urine as a phase I and two phase Ⅱ metabolisms. It is speculated that oxidation, methylation and gluconic acid conjugation reactions are the main metabolic pathways of vitexin in rat in vivo, and gluconic acid conjugation is a strong metabolic species.