In this study, a simple and efficient procedure for preparation of three flavone aglycones (luteolin, apigenin and genkwanin) from Equisetum palustre L. was developed using macroporous resin followed by gel resin flash chromatography. Among six widely used macroporous resins, D101 resin was chosen to enrich three flavone aglycones because of its good enrichment efficiency in static and dynamic tests. After one run treatment with D101 resin, the contents of luteolin, apigenin and genkwanin were 13.3-fold, 12.5-fold and 12.9-fold increased with recovery yields of 83.6%, 78.8% and 81.2%, respectively. The enriched sample was directly subjected to flash chromatography on Toyopearl HW-40S gel resin column. Luteolin, apigenin and genkwanin with purities of more than 97% were produced with recovery yields of 91.8%, 92.3% and 93.1% by one gel resin flash chromatography run. The developed procedure boasts easily reusable solvents, production of high-purity product as well as high recoveries, and allows an easy scale-up. Therefore, it is a promising basis for large-scale preparation of flavone aglycones from Equisetum palustre or other plant extracts. (C) 2013 Elsevier B.V. All rights reserved.