New methods are presented for characterizing MIPs. These methods address the problems of quantitatively comparing the binding properties of different MIPs. Heterogeneous binding models were applied to MIPs based on an exponentially decaying distribution known as the Freundlich isotherm. The Freundlich isotherm was found to accurately model the binding isotherm of the majority of non-covalently imprinted MIPs. Using this model the experimental binding isotherm can be fit in log-log form to a linear equation from which the fitting parameters can be used to plot a quantitative affinity distribution which is a plot of the number of sites with respect to the binding constant of those sites. Comparison of MIPs using this methodology allowed for simpler and more accurate assessment of the binding properties than by previous methods such as the limiting slopes analysese of curved Scatchard plots.

In Molecular Imprinting the nature of the templated binding sites and the mechanism of their formation are still poorly understood. For this reason our groups are carrying out fundamental studies concerning known imprinting protocols, with the primary aim of shedding light on the role of the template in the different steps of the polymerisation, from the formation of primary chains to the build-up of the porous structure. In this paper we report our initial results concerning copolymers of methacrylic acid (MAA) and ethyleneglycol dimethacrylate (EDMA) and their formation in presence or absence of the templates 9-ethyladenine, ametryn or terbutylazine. Monitoring the monomer disappearance by 1H-NMR showed that the presence of templates such as 9-ethyladenine significantly retarded the polymerisation but otherwise had minor influence on the relative reactivity of the monomers. The latter appeared in most cases to be stoichiometrically incorporated into the polymer. The signals arising from the template experienced little or no shift in the early stage of the polymerisation, although pronounced broadening was observed. By delaying the addition of the template, it was observed that binding sites with high selectivity could be induced more than one hour after the gel point of the system had been passed. Finally, the results of post-polymerisation curing on the dry and swollen state porosities and the recognition properties of terbutylazine imprinted polymers are reported. This treatment when performed at temperatures between 100-120°C, slightly enhanced the selectivity of the polymers, whereas at higher temperatures the polymers lost their molecular recognition properties. Swollen state porosity derived from inverse size exclusion chromatography (ISEC) revealed an interesting sharpening of the pore size distribution for the imprinted compared to the non-imprinted polymers.