Column selection for comprehensive two-dimensional liquid chromatography using the hydrophobic subtraction model

Abstract Two-dimensional (2D) liquid chromatography (2DLC) methods have grown in popularity due to their enhanced peak capacity that allows for resolving complex samples. Given the large number of commercially available column types, one of the major challenges in implementing 2DLC methods is the selection of suitable column pairs. Column selection is typically informed by chemical intuition with subsequent experimental optimization. In this work a computational screening method for 2DLC is proposed whereby virtual 2D chromatograms are calculated utilizing the Snyder–Dolan hydrophobic subtraction model (HSM) for reversed-phase column selectivity. Towards this end, 319 225 column pairs resulting from the combination of 565 columns and 100 sets of 1000 diverse analytes are examined. Compared to other screening approaches, the present method is highly predictive for column pairs that are able to resolve the largest number of analytes. This approach shows a strong sensitivity to the choice of the second dimension column (having a shorter operating time) and a preference for those with embedded polar moieties, whereas a relatively weak preference for C 18 and phenyl columns is found for the first dimension.