Abstract In this study, the adsorption/desorption characteristics of quercetin, luteolin and apigenin from Flos populi extract (Populus tomentosa Carrière, Salicaceae) on twelve macroporous resins (NKA-9, HPD-600, HPD-826, HPD-750, HPD-400, DM-130, AB-8, SP-825, X-5, D-101, HPD-100, HPD-200) were evaluated. Both high adsorption and desorption capacities of quercetin, luteolin and apigenin from Flos populi extract on SP-825 resin indicated that SP-825 resin was appropriate and its data were well fitted to the Langmuir and Freundlich isotherms. To get the optimal separation process, the influences of factors such as flow rates, loading sample volumes, concentrations of desorption solution were further investigated. Column packed with SP-825 resin was used to perform dynamic adsorption and desorption experiments. After one round of treatment, the contents of quercetin, luteolin and apigenin in the final products were 3.75-fold, 3.67-fold and 3.54-fold increased with recovery yields of 87.25, 85.19 and 82.22%, respectively. The results showed that the preparative enrichment of quercetin, luteolin and apigenin was available via adsorption and desorption on SP-825 resin. This method is a promising basis for the large-scale preparation of quercetin, luteolin and apigenin from Flos populi.