Most developments in column and instrument technology over the last 10–15 years have been focused on achieving higher separation efficiencies in a shorter time (UHPLC), with smaller particles and higher pressures. Although further decreases in particle size and/or increases in operating pressure are theoretically feasible, the potential gain is limited. On the other hand, samples are becoming more and more complex in terms of number of components and chemical heterogeneity, particularly in metabolomics, lipidomics and the analysis of biopharmaceuticals. Often, a single method is not enough to fully separate all components in a mixture and the use of different (orthogonal) separation mechanisms is required.