Abstract In this study, antioxidant peptides were prepared from hairtail surimi by hydrolysis. Response surface methodology coupled with a Box-Behnken design was applied to optimize hydrolysis conditions. The optimum conditions were determined as: 12.1 h incubation time, 44.74 °C incubation temperature and enzyme concentration 1858.85 U/g. The amino acid compositions analysis showed most abundant amino acid in hairtail surimi antioxidant peptides (HAP) was Glutamic acid followed by Aspartic acid and Lysine. In addition, the fraction of HAP with a molecular weight less than 3 kDa showed the strongest antioxidant activity among the 4 fractions obtained by ultrafiltration. Furthermore, HAP showed good stability in mild acidic, alkaline, or salt solutions, in in vitro digestive juice, and upon heating. In conclusion, hairtail surimi might be a good source to produce antioxidant peptides, which can be used as a natural antioxidant in food industry.