The polysaccharides of Flammulina velutipes were isolated in hot water,and precipitated with alcohol.The protein in polysaccharides was removed by Sevage way.The polysaccharides were graded to A1 and A2 with DEAE52 column chromatography,and then composition A2 was showed to be the single composition with Sephadex S-200 column chromatography.The anti-oxidation experiments indicated that the polysaccharide of A2 had anti-oxidation activity and this activity was increased with the increased concentration of the polysaccharide.The results of this study can provide the theory foundation for the further development and utilization of Flammulina velutipes.