Abstract: By comparing static adsorption properties of flavonoids from the Oenanthe javanica extraction on six types of macroporous adsording resin, AB-8 had a higher adsorption capacity. The dynamic adsorption separating condition including loaded rate, ethanol concentration and amount of wash solvent, desorption velocity were studied. Results indicated that static adsorption rate for flavonoid from the Oenanthe javanica was71.37%, desorption rate was 76.57%. The optimized condition was loading rate 1mL/min, desorption rate 1.5mL/min, 4BV 80% ethanol as eluent.Under this condition the purity of the Oenanthe javanica extracts with 42.60mg flavone content was improved from 7.1% to 53.78% and the enrichment amount of AB-8 macroporous adsording resin was 7.6 multiple. High performance liquid chromatography (HPLC) analysis results showed that the flavonoids of Oenanthe javanica extracts before and after purified both included rutin, quercetin, quercetin-3-O-glucoside, kaempferol. Results of HPLC analysis indicated that the purity of Oenanthe javanica flavonoids was improved from 6.44% to 55.65% and the enrichment amount of AB-8 macroporous adsording resin was8.65 multiple.