Abstract: To establish a method for the Rosavin of isolation and purification, the result of process conditions were as follows:the conditions of HPLC test is acetonitrile-0.2% phosphoric acid ( 65/43, v/v) of mobile phase, and column temperature condition 30 ℃; on the comparison of six resin's adsorption and the establishment of the resin were preliminary isothermal static adsorption curve, we can determined the optimal resin was D312 resin, and the largest loading concentration was0.24 mg/m L; column height 30 cm, sample size 4 BV, flow rate 1 ~ 2 BV/h. The results showed that the Rosavin content had increased from 3.00% to 68.76%, based on the consideration of four factors in column chromatography process. The macroporous adsorption resin of isolation and purification of Rosavin is reasonable, it is suitable for industrial applications.