Abstract: A novel molecularly imprinted polymer (MIP) was prepared by thermal polymerization with 17β-estradiol as the template, 2-(methacryloyloxy)ethyl phosphate as the functional monomer, trimethylolpropane trim ethacrylate as the crosslinker, azodiisobutyronitrile as the initiator, and a mixture of acetonitrile and toluene (3+1)as the porogen.The bonding mechanism and identifying characteristics of the MIP were evaluated by UV, NMR and molecular simulation, which indicated that the obtained MIP interacted specifically with 17β-estradiol with a ratio of 1∶2 by hydrogen bonding to form complexes. The IR, SEM, porosity analysis, TG and Scatchard plot further indicated that there was pore structure in the MIP with some functional groups and spatical action sites which could interact with the template molecular.