Resveratrol in the aqueous extraction of Polygonum cuspidatum was separated and purified by liquid extraction assistance macroporous adsorption resin mixed bed. Firstly, petroleum ether and ethyl acetate were used to replace diethyl ether that was an effective solvent for separate Resveratrol from aqueous extract. Then, Resveratrol was further separated by macroporous adsorption resin-mixed bed. Finally, the experimental conditions were optimized. When the volume ratio of water and organic mixture was 2/3, extraction time interval was 60 min and extraction times was 5 times, the recovery rate of Resveratrol was 81.10% and the purity was 87.12%.