Simulation of size exclusion chromatography for characterization of supramolecular complex: a theoretical study.

A simple chromatographic model was used to simulate the retention behavior of supramolecular complex in size exclusion chromatography (SEC). Two fundamental model complexes, directional complementary couple (AB) and self-complementary unit (AA), were employed to investigate the influence of various parameters in the SEC characterization/confirmation of supramolecular association. Peak tailing, peak fronting, peak splitting and retention time shifting were observed under different conditions. It was found that the chromatographic peak shape and retention time were strongly affected by the association constant, sample concentration, as well as the addition of a reactant in the mobile phase. Furthermore, using the same model, the chromatographic process of Hummel-Dreyer procedure was simulated, and the results indicated that the procedure can be a good method for the determination of association constants for AB type complexes.