Chapter 2 Reversed-phase and hydrophobic interaction chromatography of carbohydrates and glycoconjugates

Abstract

This chapter discusses the fundamentals of reversed-phase chromatography (RPC) and hydrophobic interaction chromatography (HIC), describes the chromatographic systems—for example, the mobile and stationary phases, and reviews the most useful approaches and concepts that are developed for the RPC and HIC of carbohydrate species. RPC and, to a lesser extent, HIC are widely used in the separation and determination of carbohydrate species. In RPC and HIC, solute retention has the same intrinsic physicochemical basis—the hydrophobic effect that is essentially a solvent effect. In both RPC and HIC, the non-polar moieties of the solute and the stationary phase undergo noncovalent association in hydro-organic media or aqueous solutions of neutral salts, respectively. In contrast, the operation conditions, as well as the nature of the mobile and stationary phases used in HIC and RPC, are very different. In RPC, a stationary phase is a nonpolar surface consisting mainly of long alkyl chains covalently bound to solid support.