High performance liquid chromatographic analysis of carbohydrates is a real challenge; because these compounds exhibit similar chemical and physical properties, they are more difficult to analyze than most other classes of compounds, to date no single chromatographic column or method being capable of separating all carbohydrates. Choosing the best column for carbohydrates analysis requires consideration of stationary phase chemistry, retention capacity, particle size and column dimensions. This research focuses on mono- and di-saccharides analysis, represented here by arabinose, fructose, glucose, saccharose and maltose, different commercially available columns being tested; isocratic separations were achieved using as mobile phase different acetonitrile / water mixtures at 1.2 mL min-1. A test carbohydrate mixture was used and the resulted separations were compared in order to select the best suited column.