Objective:The aim of this work is to obtain the optimal purification process of pigment from Boletus edulis by macroporous resin, and study its antioxidant activity. Methods:Based on the static and dynamic test, the influence of resin type, sample concentration, pH, loading flow ratio, ethanol concentration and elution flow ratio on the adsorption and desorption activity of Boletus edulis pigment were investigated to determine the optimal purification conditions. Then the structure, the DPPH·, ABTS+· and ·OH scavenging activity of the Boletus edulis pigment purified was also evaluated. Results:AB-8 was the most suitable type of macroporous resin and the optimum purification conditions were as follows: the pigment concentration of 1.5 mg/mL, pH of 2.0, the sample flow ratio of 2.0 mg/mL, ethanol concentration of 70% (V/V) and the elution flow ratio of 2.0 mL/min. Under these conditions, the purification efficiency of Boletus edulis pigment was 269%. The IC50 of the final purified product for scavenging DPPH·, ABTS+· and ·OH were (0.081±0.001), (0.017±0.011) and (0.119±0.001) mg/mLrespectively. Especially, the ·OH scavenging ability was higher than VC. Conclusion:AB-8 resin was suitable for separation and purification of Boletus edulis pigment, and the pigment after purification revealed strong antioxidant activity.