This document introduces the application of Agilent Hi-Plex columns, which are mainly used for the separation of carbohydrates, alcohols, and organic acids. These columns use ion-exchange ligand-exchange columns, suitable for the analysis of compounds such as simple sugars, alcohols, oligosaccharides, and organic acids in foodstuffs. The columns use isocratic conditions with water as the eluent and temperature as the main variable for control of resolution. When samples contain organic acids, especially in the presence of neutral mono- and disaccharides, Agilent Hi-Plex H is the preferred option. The column is run using water as the eluent for the analysis of sugars and organic acids or, more commonly, dilute acid for separation.