To investigate the hypolipidemic effects and antioxidant levels of total flavonoids from seabuckthorn fruit peel and residue, in vitro and in vivo experiments were conducted to examine the hypolipidemic functionality of the purified total flavonoids from seabuckthorn fruit peel and residue (TFSE). The antioxidant activity of TFSE was determined through in vitro experiments, and its binding capacity to three types of bile acids was investigated by simulating the human gastrointestinal environment. This was conducted to evaluate its antioxidant and hypolipidemic effects in vitro. In the in vivo experiment, a high-fat diet-induced hyperlipidemia model was established in C57BL/6J mice, and TFSE was administered orally to compare its effects on body weight, serum, hepatic lipid levels, liver function indicators, and oxidative indicators. Then, histopathological observations of the liver tissue were completed. The in vitro study showed that TFSE exhibited strong scavenging abilities against DPPH and ABTS+ radicals, with IC50 values of 5.20 and 10.24 μg/mL, respectively, thus indicating effective antioxidant activity. Moreover, TFSE showed strong binding capabilities with sodium cholate, glycocholate, and taurocholate, with IC50 values of 11.11, 2.61, and 3.50 mg/mL, respectively, thus highlighting its binding solid capacity with bile acids. Results of the mice experiments showed that, compared to the model group (HFD), the L-TFSE and H-TFSE groups showed inhibited body weight gain, whilst (P © The Author(s) 2024.