Determination of adsorption isotherms by means of HPLC: adsorption mechanism elucidation and separation optimization

The purpose of this review is to illustrate the most important techniques for isotherm determination by means of HPLC. Starting on the traditional Frontal Analysis approach, Frontal Analysis by Characteristic Point, Elution by Characteristic Point, Perturbation Method in its different applications will be considered to conclude with the most recent Inverse Method approach. Since many of these techniques are based on the fundamentals of nonlinear chromatography, a short overview of the theory of nonlinear chromatography is presented. Emphasis is given to the most recent applications of these techniques for pharmaceutical applications, characterization of binding mechanisms, bioaffinity studies, molecular and chiral recognition processes.