Abstract: The objective of this study was to extract and purify antioxidant substances extracted from Picea koraiensis by alcohol and evaluate their antioxidant activity in vitro. First, antioxidant substance was extracted by different concentrations of ethanol and purified by macroporous resin D101. Then, different samples were collected and the antioxidant activity of the components with the higher content of dry matter and polyphenols was evaluated. Four antioxidant parameters (the capacity of scavenging hydroxyl and DPPH radicals, total reducing power and lipid peroxidation inhibition effect) were measured. The results indicated that PT40%-E showed the highest activity of scavenging hydroxyl radical (scavenging rate and the value of IC50 were 97.998% and 119.101g/mL, respectively). PT20%-D showed the highest activity of scavenging DPPH radical (scavenging rate and the value of IC50 were 99.617% and 9.009g/mL respectively). Then, PT80%-D showed higher ability of total reducing and the absorbance value was 2.209 when the concentration was 300g/mL. Finally, PT40%-D group showed the highest lipid peroxidation inhibition effect (the inhibition ratio and the date of IC50 were 99.148% and 271.872g/ mL respectively ). The correlation analysis between the antioxidant activity and the content of polyphenols, proanthocyanidins and flavonoids showed that the polyphenol was major contributor to antioxidant properties.