Molecular crowding is a new approach promoting molecular imprinting more efficiently.In this work, this concept was applied to the preparation of low cross-linked imprinted polymers in the presence of an immobilised template for stabilizing binding sites and improving molecular recognition.An imprinted monolithic column was synthesized using a mixture of 2,4-diamino-6-methyl-1,3,5-triazine and 2,4-diamino-6-(methacryloyloxy) ethyl-l,3,5-triazine as templates, methacrylic acid as functional monomer and ethylene glycol dimethacrylate as cross-linker.Some polymerization factors, such as template-monomer molar ratio, the composition of the porogen and crosslinking density, on the imprinting effect of resulting MIP monolith were systematically investigated.The results indicated that the imprinted monolithic columns prepared in the presence of molecular crowding agent retained affinity and specificity for template even when prepared with a level of cross-linker as low as 9%.Moreover, a stoichiometric displacement model for retention (SDM-R) was successfully applied to evaluate the interaction between the solute and the stationary phase.Compared with the low cross-linked MIP prepared by conventional polymerization, the molecular crowding-based low cross-linked monolithic MIPs showed higher selectivity.