Comparison of performance of high‐performance liquid chromatography columns packed with superficially and fully porous 2.5 μm particles using kinetic plots

Abstract

A recently introduced 2.5 μm fully porous support (Kromasil Eternity) is compared with three different brands of superficially porous material (Kinetex, Halo and Poroshell 120) by means of the kinetic plot method using pharmaceutical compounds from GlaxoSmithKline as probe molecules. The kinetic plot method immediately shows the range of plate numbers wherein a support performs better than another. Results from experiments carried out at pH 4.5 and 8.0 are presented in order to assess the pH stability of the tested phases. Moreover, since all supports are able to withstand pressures higher than 400 bar, they have been evaluated both on HPLC and UHPLC instrumentation. True average particle sizes were determined by SEM images taken from loose stationary phases. Kinetex outperforms the other columns in HPLC conditions for practically relevant efficiencies, but shows poor packing quality in the 100×2.1-mm format. Kromasil is advantageous for simple and fast separations on short columns both in HPLC and in UHPLC conditions. Halo achieves the highest efficiencies of all columns at the lowest pressure cost and shows a noticeable lower axial diffusion. Poroshell 120 has the best packing quality reproducibility across the tested formats. All columns preserve their performance at high pH.