Evaluation of retention behavior of carbohydrate compounds on the zwitterionic hydrophilic interaction liquid chromatographic column

Abstract

Because of the strong polarity of carbohydrate compounds which have weak retention in reverse-phase liquid chromatography (RPLC) , hydrophilic interaction liquid chromatography (HILIC) is suitable for the separation and analysis of the carbohydrate compounds. In this study, nine carbohydrate compounds were selected as the test probes to systematically evaluate the retention behavior of the carbohydrate compounds on a Click TE-Cys column with zwitterionic stationary phase. And the effects of the ratio of organic phase and salt concentration in mobile phase on the retention behavior were investigated as well. The experiments proved that the nine carbohydrate compounds could be eluted in the order of their polarities from the Click TE-Cys column. With the increase of organic phase ratio, the retention of carbohydrate compounds was enhanced. As the increase of salt concentration, the retention of the carbohydrate compounds increased except sialic acid. Using displacement-adsorption liquid phase interaction model to simulate the retention behavior of the carbohydrate compounds under HILIC, the retention behavior of HILIC was described by the retention equation of ln k = a + bln C(B) + cC(B), and the retention values of HILIC were analyzed by multiple linear regression. The results demonstrated that the retention behavior of carbohydrate compounds on the Click TE-Cys was in accordance with the retention regularity of HILIC.