Molecularly imprinted polymers (MIPs) with template-selective recognition sites and incorporated fluorosensor were prepared against adenosine 3',5'-cyclic monophosphate (cAMP). The time-resolved fluorescence decay analysis was used to investigate the specificity and affinity of the binding of template molecules to the MIP. The fluorescence decays were modelled in terms of lifetime distributions and two fluorescence lifetimes were observed for the MIPs. The lifetime distributions are interpreted in terms of the heterogeneity of the functionalised imprinted cavities. Quenching of fluorescence of the imprinted polymer with increasing concentration of aqueous CAMP was observed from the fluorescence lifetime parameters data. The mechanisms of interactions between the CAMP and fluorosensor molecules inside the imprinted cavity in comparison with the interactions in solution are discussed.