Separation of sugars by ion-exclusion chromatography on a cation-exchange resin

A method for the separation of N-acetylmannosamine and N-acetylglucosamine is described, which consists of chromatography of the two sugars on a column (30 x 1 cm) of the cation-exchange resin, Dowex 50W-X2, in borate buffer at pH 7.8. N-Acetylmannosamine is eluted near the void volume, while N-acetylglucosamine emerges in a more retarded position. It is postulated that the separation occurs as a result of the combined effects of ion exclusion and gel permeation. Thus, in borate solution, N-acetylmannosamine presumably exists largely as a negatively charged complex and is therefore excluded from the sulfonated polystyrene matrix, while N-acetylglucosamine occurs mainly as the free sugar in the equilibrium mixture and, being a neutral compound, has free access to the porous resin. The proposed mechanism for the separation was supported by the finding that glucose and glucose 6-phosphate could also be separated on a column of the same resin, with water as the eluent.