OBJECTIVE: To explore potential biomarkers of acute hepatic injury induced by diosbulbin B and study its hepatotoxicity mechanism from metabolic aspect. METHODS: Totally 24 mice were randomly divided into normal group (0.5% sodium carboxymethylcellulose solution) and diosbulbin B group (200 mg/kg), with 12 mice in each group. They were given relevant medicine intragastrically; 24 h later, the levels of ALT, AST in serum and MDA, SOD in liver tissue were determined. HE staining was used to observe the histopathological changes of liver tissue. The levels of metabolism products in urine and liver tissue were determined by 1H-NMR. The potential biomarkers of diosbulbin B hepatotoxicity were screened by OPLS-DA. RESULTS: Compared with normal group, the levels of ALT and AST in serum, the level of MDA in liver tissue were increased significantly in diosbulbin B group, while the level of SOD in liver tissue was decreased (P<0.05). The liver cells were damaged,with marked pathological changes. The levels of metabolic products were significantly changed in urine and liver tissue. Three and six potential biomarkers related to acute hepatic injury were obtained (P<0.05), i.e. α-ketopglutaric acid, dimethylamine, trimethylamine oxide (urine) and glutathione, taurine, inosine, tyrosine, phenylalanine, histidine (liver tissue). CONCLUSIONS: The hepatic injury induced by diosbulbin B are caused by free radical oxidation. The mechanism of hepatotoxicity may be the effect of diosbulbin B on tricarboxylic acid cycle, amino acid metabolism and intestinal flora.