Effect of temperature on the retention of amino acids and carbohydrates in high-performance anion-exchange chromatography.

Effect of temperature on the retention of amino acids and carbohydrates in high-performance anion-exchange chromatography (HPAEC) was investigated. Amino acids retention exhibited both exothermic and endothermic behavior. Significant selectivity variations of some weakly retained amino acids were achieved through changing temperature. However, temperature was almost ineffective at changing the selectivity of strongly retained amino acids. Sugars retention exhibited only exothermic behavior. However, individual carbohydrates showed different relative changes in retention time with increased temperature. There is the general regularity, which is the variation of retention time increases as the sequence of monosaccharide, disaccharide, trisaccharide and tetrasaccharide when the column temperature is increased. Selectivity changes of carbohydrates, including elution order reversal, were also achieved by varying temperature. Temperature was effective at changing the selectivity of between amino acids and carbohydrates. In achieving optimal separation of amino acids and carbohydrates, temperature is a valuable tool.