Retention of ionizable compounds in reversed-phase liquid chromatography. Effect of the ionic strength of the mobile phase and the nature of the salts used on the overloading behavior.

Abstract

The retention mechanism of the protonated cation in propranolol chloride on C18-Xterra was investigated using mobile phases of various compositions. Accurate adsorption data were measured by frontal analysis, with a mixture of methanol and water (25% methanol), with no salt, as the mobile phase. The experimental isotherm has at least two inflection points, at concentrations of about 0.2 and 6.0 g/L, respectively. This precludes the modeling of these data with a simple convex-upward isotherm (e.g., Langmuir). The adsorption energy distribution or relationship between the number of sites on the adsorbent surface and the energy of adsorption on these sites was calculated by assuming Moreau isotherm behavior (S-shaped isotherm). This model has never been applied to describe the surface heterogeneity of any RPLC adsorbent. The calculation converged toward a bimodal energy distribution. Accordingly, the bi-Moreau model is the simplest theoretical model accounting for the adsorption data of propranolol from a mobile phase without salt. The complex-overloaded band profiles of propranolol measured in the presence of increasing concentrations of a supporting salt (KCl) in the mobile phase demonstrate that the same isotherm model applies also under these conditions, as was merely assumed in a previous work. The elution band profiles of propranolol calculated with the bi-Moreau isotherm model for solutions of salts of different natures (CaCl2, CsCl, Na2SO4) in the same mobile phase agree very well with the experimental band profiles.