The flavonoids from stems and leaves of Physalis alkekengi ( PSF ) were refined by macroporous adsorption resin, and their antioxidant activities were evaluated in vitro and in vivo. The PSF were obtained using with AB-8 macroporous adsorption resin washed by 70% ethanol, and the purity was 39.16%. After refining, the scavenging activities of PSF on NO^-₂·and ABTS⁺· were 81.36% and 91.49%. The anti-oxidative capability in linoleic acid lipid overoxidation system was 85.26%. The refined PSF not only significantly increased thymus index and spleen index（P＜0.05）, but also increased the activity of T-AOC, GSH-Px and SOD in serum（P＜0.05） and decreased the content of MDA in serum（P＜0.01）. The refined PSF showed strong antioxidant activity in vitro and in vivo.