The aim of this study was to optimize and characterize Flavourzyme hydrolysis conditions for the preparation of antioxidant peptides from duck meat, using response surface methodology. The results indicated that optimal Flavourzyme hydrolysis conditions for preparation of antioxidant peptides from duck protein were a temperature of 50.19 °C, pH 5.45, and a reaction time of 1.03 h. Compared to non-hydrolyzed duck meat, Flavourzyme hydrolysis significantly improved the hydroxyl-radical scavenging, DPPH radical-scavenging, ferrous ion-chelating, reducing, and ABTS radical cation-scavenging activities of duck meat. Therefore, Flavourzyme can be regarded as an effective hydrolytic enzyme for the preparation of antioxidant peptides from duck meat.