Abstract: In the present study, the interactions of components in a (-)- nicotine molecular imprinting polymerization mixture have been studied by H-1 NMR spectroscopy. The dissociation constants for complexation of template by a functional monomer analogue, acetic acid, have been determined. Nicotine was shown to self- associate at concentrations comparable to those used in previous molecular imprinting studies (app K-diss = 0.082 M in CDCl3 at 298 K). The extent of self-association was enhanced by the presence of acetic acid. Previous studies on (-)-nicotine- imprinted methacrylic acid-ethylene dimethacrylate co-polymers suggested the involvement of recognition sites for template- template complexes. Collectively these results provide the first direct evidence for the presence of template-template complexes, and support the previously hypothesized basis for cooperative ligand recognition events in this polymer system. (C) 2003 Elsevier B.V. All rights reserved