Antioxidant peptides were prepared from hairy antler grounds by high-intensity pulsed electric fields (PEF) jointed bromelain, and then they were isolated and purified. The degree of hydrolysis and scavenging rates of DPPH· was considered as test indexes. According to the results of the single factor experiment, the response surface methodology (RSM) experiment was conducted on three factors at five levels to analyze the factors and interaction effects on the scavenging rates of DPPH·. Then the macroporous resin was used to desalt, separate and purify hairy antler antioxidant peptides, and the antioxidant activities of hairy antler peptides were also analyzed. The optimal conditions of enzymatic hydrolysis are as follows：the substrate concentration 15%, bromelain concentration 7.04 U/mg, electric filed intensity 17.3 kV/cm, pulse number 7, the scavenging rates of DPPH· by hairy antler antioxidant peptides was (74.76±2.03)%. The scavenging rates of DPPH·, ·OH and O^-₂· of 75% ethanol elution component C3 were the highest, and the antioxidant activity of elution component C3 sharply increased compared with crude peptide, when hairy antler peptides were featured with a considerable antioxidant activity, and hairy antler could be a potential source of preparing peptide with the strong antioxidant activity.