Taking polygonatum extraction as raw material, the adsorption rate and desorption rate of total flavonoids as the indexes, the type of macroporous resin was screened out by dynamic adsorption - desorption method. The process condition of absorbing the total flavonoids of polygonatum by D-101 resin was established by single factor experiments and orthogonal experiment; the main factors, concentration and dosage of eluent, was discussed. The optimal adsorption process condition was: the amount of macroporous resin 55g, the concentration of upstream solution 40.54μg/mL, pH 6, and adsorption velocity 0.5BV/h. During the process of desorption solution, the concentration of ethanol was 60% and the dosage of ethanol was 100mL. After being purified by D-101 macroporous resin, the purity of total flavonoids was increased from 0.36％to 2.05%. This purification method is cheap, safety, simple and has a high application value.