In this study, the ideal adsorbent for Dendrobium hu-oshanense polyphenols were obtained, after the comparison test for the adsorption-desorption properties of different types of resins. The purification parameters were optimized by adsorption and desorption tests. The antioxidant activities of polyphenols were evaluated in vitro. The results showed that AB-8 macroporous resin was considered as suitable material for purifying and separating Dendrobium huoshanense polyphenols.And the macroporous resin reached equilibrium within 150 min. The optimal processing parameters were as follows: an initial polyphenol concentration of 2.0 mg/mL, pH 4.0, a flow rate of 1.0 mL/min for feeding and a flow rate of 1.0 mL/min for eluting. The total polyphenols purity reached 76.2%. Four groups of polyphenol components were obtained by dynamic elution with different ethanol concentrations, which were named DHP-1, DHP-2, DHP-3 and DHP-4, respectively. The contents of them were 16.3%, 14.8%, 20.9% and 26.2%, respectively. The total antioxidant capacity of them indicated that DHP-3＞DHP-4＞DHP-1＞DHP-2＞crude polyphenols.