Flavonoids and bonito protein hydrolysates were prepared respectively using rhizoma smilacis glabrae and bonito fish as raw materials. The Flavonoids from rhizoma smilacis glabrae was screened with the index of flavone extraction rate， 1，1-diphenyl-2-picrylhydrazyl free radical （DPPH·） scavenging activities and xanthine oxidase （XOD） inhibitory activity. The hydrolytic enzyme was optimized with the index of protein recovery rate， DPPH· scavenging activities， XOD inhibitory activity and the amino acid analysis of the optimal bonito protein hydrolysates. The effect of interaction between flavonoids extracts from rhizoma smilacis glabrae and bonito protein hydrolysates on antioxidant activity and XOD inhibitory activities was examined. Then investigation the mechanism of the interactions of flavonoids and peptide using fluorescence spectroscopy. The results demonstrated that flavonoids extracted by ethanol showed higher flavonoids recovery rates， DPPH· scavenging activities and XOD activity than aqueous extract. The optimum hydrolytic enzyme of bonito fish is papain. It was observed that the XOD activity of flavonoids-peptide complexes was significantly increased， and the antioxidant capacity showed significant synergistic effect. DPPH· scavenging activities of flavonoids-peptide complexes was slightly improved in pasteurization， while significantly decreased in high temperature sterilization. At pH 6.0， DPPH· scavenging activities was greater than that of pH 5.5 and 6.5. After fluorescence spectrum analysis， flavonoids may through hydrogen bonding with bonito protein hydrolysates to form new compounds.