OBJECTIVE: To study the pharmacokinetics and absolute bioavailability of oleanolic acid and its derivative dihydrooleanolic acid in rats in vivo. METHODS: 12 SD rats were randomly divided into oleanolic acid administration group and dihydrooleanolic acid administration group, 6 in each group. All rats were intragastrically given related medicine (50 mg/kg), then intravenously injected related medicine (2 mg/kg) in tail vein after 1 week. Sample blood 0.3 mL was taken from tail vein before administration and after administration (0.1, 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 5, 7, 9, 12 h for ig; 0.05, 0.15, 0.25, 0.5, 0.75, 1, 2, 3, 5, 7, 9, 12 h for iv). UPLC-QTOF was conducted to determine the plasma concentration, and DAS 2.0 pharmacokinetic software was used to calculate pharmacokinetic parameters and absolute bioavailability. RESULTS: After ig and iv oleanolic acid, the AUC0-∞ were (232.10±7.17), (1 203.99±19.65) ng·h/mL, t1/2 were (1.75±0.10), (1.41±0.04) h respectively; after ig, cmax was (121.3±18.92) ng/mL, tmax was (0.54±0.10) h, absolute bioavailability was 0.77%. After ig and iv dihydrooleanolic acid, AUC0-∞ were (382.03±23.73), (386.14±10.65) ng·h/mL, t1/2 were (2.47±0.45), (1.44±0.03) h; after ig, cmax was (124.52±12.28) ng/mL, tmax was (0.63±0.14) h, absolute bioavailability was 3.96%. CONCLUSIONS: The absolute bioavailability of dihydrooleanolic acid is significantly higher than oleanolic acid in rats.