Effect of the mobile phase composition on the isotherm parameters and the high concentration band profiles in reversed-phase liquid chromatography.

Single-component adsorption isotherm data were acquired by frontal analysis for phenol on a C18-Kromasil packed column, under reversed-phase liquid chromatography conditions, using various methanol-water solutions (30-60%, v/v, methanol). The isotherm model accounting best for these data was the biLangmuir model. With increasing methanol content, the two saturation capacities decrease, particularly that of the high-energy sites, the adsorption constant of the low-energy sites decreases significantly and that of the high-energy sites decreases strongly. These results allow a quantitative investigation of the properties of the high-energy sites (which are not necessarily the so-called active sites), a feature rarely discussed yet. The band profiles calculated with the numerical values of the isotherm model parameters derived by fitting the frontal analysis data to the model and using the equilibrium-dispersive model agree very well with the experimental band profiles in the whole concentration range.