Prediction of peak shape as a function of retention in reversed-phase liquid chromatography

Optimisation of the resolution of multicomponent samples in HPLC is usually carried out by changing the elution conditions and considering the variation in retention of the analytes, to which a standard peak shape is assigned. However, the change in peak shape with the composition of the mobile phase can ruin the optimisation process, yielding unexpected overlaps in the experimental chromatograms for the predicted optimum, especially for complex mixtures. The possibility of modelling peak shape, in addition to peak position, is therefore attractive. A simple modified-Gaussian model with a parabolic variance, which is a function of conventional experimental parameters: retention time (tR), peak height (H0), standard deviation at the peak maximum (sigma0), and left (A) and right (B) halfwidths, is proposed. The model is a simplification of a previous equation proposed in our laboratory. Linear and parabolic relationships were found between the peak shape parameters (sigma0), A and B) and tR, with a mean relative error of 1-5% in most cases. This error was partially due to variations in peak position and shape among injections, which in some cases were above 2%. Correlations between (sigma0, A and B) and the retention time, which is easily modelled as a function of mobile phase composition, allowed a simple and reliable prediction of chromatographic peaks. A parameter that depends on the slopes of the linear relationships for A and B versus tR is also proposed to evaluate column efficiency. The modified-Gaussian model was used to describe the peaks of six diuretics of diverse acid-base behaviour and polarity, which were eluted with 15 mobile phases where the composition was varied between 30 and 50% (v/v) acetonitrile and the pH between 3 and 7.