In this study, a simple rand efficient protocol is developed for preparation of biochanin A and genistein from Dalbergia odorifera T. Chen leaves using macroporous resin followed by flash chromatography. The adsorption and desorption capacity of 14 macroporous adsorption resins for biochanin A and genistein were evaluated, and AL-2 resin showed better properties for biochanin A and genistein. After treatment with AL-2 resin, the contents of biochanin A and genistein in the enriched product were 6.60-fold and 6.41-fold increased with recovery yields of 87.13% and 84.60%, respectively. Furthermore, the operating parameters of flash chromatography were optimized. The optimal conditions were as follows: stationary phase: silica gel, elution system: n-hexane/ethyl acetate, sample/silica gel ratio: 1.3:40 and flow rate: 50 mL/min. After one flash chromatography run, purities of biochanin A and genistein effectively reached over 95%, their recovery yields were 80.13% and 73.11%, respectively. The developed protocol was simple, efficient, scalable and economical, which represented an excellent alternative for the separation and purification of bioactive compounds from plants. (C) 2013 Elsevier B.V. All rights reserved.