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Molecular crowding is a new concept to obtain MIPs with greater capacity and selectivity by shifting the equilibrium of a print molecule reacting with functional monomers in the direction of complex formation side under a macromolecular surrounding.It is an interesting issue that the specialized interaction between imprinting molecules and MIPs is mainly caused by the complementary functional groups or the suited structures of the MIPs in the presence of crowding agent.For an understanding of the molecular recognition of MIP prepared in a surrounding of a crowding-inducing agent, a monolithic MIP with enrofloxacin imprinting was prepared by in situ copolymerization of imprinted molecule, methacrylic acid and ethylene glycol dimethacrylate in porogenic solvent, a mixture of polystyrene-tetrahydrofuran.Strong recognition ability with an imprinting factor of 3.03 for template was achieved on the monolithic MIP.The stoichiometric displacement model (SDM) for retention was used to investigate the recognition mechanism of molecular imprinting.The results from SDM showed that the molecular recognition on MIP monolith prepared in molecular crowding conditions mainly derived from promoting hydrogen bond formation between enrofloxacin and MAA other than the space effect of MIP cavity structure.