Three polysaccharide fractions (DDP1-1, DDP2-1 and DDP3-1) were successfully purified from the crude polysaccharide of Dendrobium denneanum by DEAE-Cellulose and Sephadex G-200 column chromatography. The average molecular weights (Mws) of these fractions were 51.5, 26.1 and 6.95 kDa, respectively. Monosaccharide components analysis indicated that DDP1-1 and DDP2-1 were composed of arabinose, xylose, mannose, glucose and galactose in a molar ratio of 1.00:2.82:57.11:140.82:7.76 and 1.00:1.62:1.18:77.5:7.79. DDP3-1 was composed of arabinose, mannose, glucose and galactose in a molar ratio of 1.00:1.03:8.84:2.00. On the basis of antioxidant test in vitro, DDP2-1 exhibited the highest antioxidant ability among these samples.