Objective:To design and synthesize the pueraria isoflavonoid-imprinted monolithic column by molecular modeling,and theoretically explore the mechanism of its specific molecular recognition. Methods:Genistein was selected as template,4-vinylpyridine,acrylamide and methacrylate were selected as candidate monomers. Density functional theory(DFT) was adopted to accurately predict the hydrogen bonding energy between template and different functional monomers,respectively. And then the optimal functional monomer and the appropriate ratio of monomer to template were screened out. As a result,the pueraria isoflavonoid-imprinted monolithic column was prepared by in situ polymerization method. Results:4-vinylpyridine was the optimum functional monomer and ideal ratio of monomer to template was 2∶1. Meanwhile,pueraria isoflavonoid-imprinted monolithic column was successfully prepared. Conclusion:These results provide the theoretical reference for the design and synthesis of pueraria isoflavonoid-imprinted monolithic column,and can be applied to the other non-covalent imprinting system.