Chapter 2 Reversed-Phase and Hydrophobic Interaction Chromatography of Carbohydrates and Glycoconjugates

Abstract

This chapter highlights reversed-phase chromatography (RPC) and hydrophobic interaction chromatography (HIC) of carbohydrates and glycoconjugates. In both RPC and HIC, solute retention has the same intrinsic physicochemical basis—the hydrophobic effect, which is essentially a solvent effect where the nonpolar moieties of the solute and the stationary phase undergo noncovalent association in hydro-organic media and aqueous solutions of neutral salts, respectively. The retention process in HIC and RPC involves an interaction between two nonpolar groupings: the hydrophobic moiety of the solute and the nonpolar ligand bonded to the surface of the solid support. The solvation process of the solute involves two distinct steps: (1) the creation of a cavity in the solvent, which is of the same shape and size as the incoming solute, and (2) the entrance of the solute into the cavity and the subsequent interaction with its surroundings. In both HIC and RPC, in addition to van der Waals interactions with both the stationary and the mobile phases, the solute also undergoes polar interaction with the mobile phase.