Simple and efficient preparation of biochanin A and genistein from Dalbergia odorifera T. Chen leaves using macroporous resin followed by flash chromatography

• Separation of active compound using this method was studied for the first time.
• After one run, high-purity compounds with high recovery yield were obtained.
• This method exhibits great potential for large-scale preparation of active compounds.

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.