Approaches to model the retention and peak profile in linear gradient reversed-phase liquid chromatography

The optimisation of the experimental conditions in gradient reversed-phase liquid chromatography requires reliable algorithms for the description of the retention and peak profile. As in isocratic elution, the linear relationship between the logarithm of the retention factor and the solvent contents is only acceptable in relatively small concentration ranges of modifier. However, more complex models may not allow an analytical integration of the general equation for gradient elution. Alternative approaches for modelling the retention in linear gradient elution are here proposed. Those based on the quadratic logarithmic model and a model proposed for normal liquid chromatography yielded accurate predictions of the retention time for a wide range of initial concentrations of organic modifier and gradient slopes, with errors usually below 1–2%. Based on the half-width changes of chromatographic peaks along one or more gradients, an approach is also reported to predict the peak profile with low errors (usually below 2−3%). The proposed approaches were applied to two sets of probe compounds (diuretics and flavonoids), eluted with acetonitrile–water gradients. The changes in retention and peak shape in isocratic and gradient elution are illustrated through diagrams that define triangular regions including all possible values of retention factors or peak half-widths (or widths) inside the selected working ranges.

► Semi-empirical approach to integrate the quadratic logarithmic retention model.
► Comparison of retention models in isocratic and gradient elution.
► Novel model to predict peak widths in isocratic and gradient elution.
► Highly accurate predictions for diuretics and flavonoids.
► Triangular plots to describe retention and width in isocratic and gradient elution.