The in Vitro anti-oxidative activity of pine needle flavonoids（PNF）and enzymatic hydrolysis were investigated. It is found that the DPPH radical scavenging activity of PNF is similar to that of ascorbic acid. The hydroxyl radical and super-oxide radical scavenging activity of PNF are 4 times and 2 times as that of ascorbic acid, respectively. As for reducing power, EC50 ratio of PNF to ascorbic acid is only 1/2.8. β-glucosidase is used to hydrolyze PNF. The optimal hydrolyzing conditions are 40℃ of reaction temperature，1/1000 of enzyme addition，0.6g/L of substrate concentration, and 5h of reaction time. The anti-oxidative activity of enzymatic hydrolyzed PNF is increased obviously. 