Abstract: The purpose of this paper was to explore method for purifying acteoside from Cistanche tubulosa by using macroporous resin. The most suitable resin was decided by static tests and then three parameters including eluent concentration,eluting time and eluting rate were optimized for developing dynamic eluting conditions. Results showed that resin HPD300 had the best absorption and desorption capabilities. The content of acteoside increased from 5.26% to 68.20% with recovery of 68.90% under the optimized dynamic eluting conditions——eluting with ultrapure water for 30 min after saturation adsorption,then with 20% and 50% ethanol for respectively 75 min and 60 min with flow rate 10 m L/min. The method developed provided a potential approach for industrial production of acteoside from Cistanche tubulosa for its relative high recovery and purity and low cost.