AIM: To explore the optimal separation of the total flavonoid from Radix Puerariae by selecting appropriate macroporous resins and to study systematically the factors which affect separation. METHODS: Static and dynamic adsoption desorption methods were adopted, and evaluated for separating efficiency by measuring the concentration of total flavonoid in Radix Pueraria with UV spectrophotometer. RESULTS: The SP70 macroporous resin had the best separating efficiency when the flavonoid content in the liquid was 0.5g per 1mL equivalemt to raw material. pH5~6. the volume of drug 60BV (resin bed volume) with the adsorption power 2BV/h. and the volume of 70%(mL?mL -1 ) ethanol as eluant 4BV with desorption power 2BV/h. In result. total flavonoid's purity could reach 80%. CONCLUSION: This method is simple and feasible with good effect of separation, which can meet industrial requirements.