Optimization model for the gradient elution separation of peptide mixtures by reversed-phased high-performance liquid chromatography. Application to method development and the choice of column configuration

Previous papers in this series super(1,2) have presented a general model for the separation of peptide and protein samples by reversed-phase high-performance liquid chromatography (HPLC) using gradient elution. This model begins with the general theory of band broadening in isocratic liquid chromatography as developed by previous workers, adds the corresponding theory which relates gradient and isocratic separation, and then generalizes the overall treatment so as to apply to both large and small molecules. In this paper the authors will illustrate the utility of their model for practical application and for gaining further insight into the fundamental nature of these gradient separations of peptides and proteins.