Abstract: The effect of separation of five anthraquinones in rhubarb by macroporous resin was investigated through static adsorption experiment in this paper. The sorption and desorption properties of five anthraquinones(aloe-emodin,rhein,emodin,chrysophanol,physcion)on macroporous resins including HPD-100,XDA-6,AB-8,LX-38,ADS-7 and ADS-17 had been compared. Then,Dynamic adsorption and desorption were performed on the chosen macroporous resin packed in glass columns to optimize the separation process.Results showed that,HPD-100 resin offered much higher adsorption and desorption ratios for five anthraquinones than other resins;under the optimised conditions:the ratio of column height to diameter bed 1∶8,loading concentration 3.64 mg/mL,loading volume 2.0 BV,flow rate 1.0 BV/h,85% ethanol as eluent,and elution volume 3 BV,the dynamic adsorption and desoption rate of five anthraquinones on HPD-100 were 86.3% and 85.9%,respectively. The content of five anthraquinones in the final product increased 2.88-times from 7.13% to 20.5%,with a recovery of 98.7%,ionic liquidsbmimBr could be removed from crude extract,which showed that the optimized conditions were feasible.