Separation of fucosylated oligosaccharides using high-pH anion-exchange chromatography with pulsed-amperometric detection.
Fucosylated oligosaccharides of the beta Gal(1----4)GlcNAc-, beta Gal(1----4)Glc-, and beta Gal(1----3)GlcNAc-series were chromatographed on a high-performance anion-exchange pellicular resin under alkaline conditions (pH congruent to 13). Fucosylation of either lactose, lactosamine (Type II chains), or lacto-N-biose (Type I chains) oligosaccharides markedly decreased the retention time (10-38 min) of the non-fucosylated form. The magnitude of the reduction was related to whether fucose replaced Gal [alpha Fuc(1----3)----GlcNAc], whether fucose was alpha-(1----2)-linked to Gal at the end of a chain, or whether fucose was linked in a subterminal position [alpha(1----3) or alpha (1----4)] to Gal or GlcNAc. The results suggest that the decreases in retention times of fucosylated oligosaccharides (10-38 min) is not attributable to the absence of a 6-OH in Fuc but instead to steric and substitution effects which affect the interaction of the most readily ionizable groups of Fuc (2-OH), Gal (2-OH), and GLcNAc (3-OH) with the stationary phase. We show that high-pH anion-exchange chromatography can effectively separate 1----2, 1----3, and 1----4 fucose positional isomers in a single chromatographic step.
Department of Biology, Johns Hopkins University, Baltimore, MD.
M R
MR
Hardy
R R
RR
Townsend
D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002852:Chromatography, Ion Exchange / Q000379:methods; D005643:Fucose / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D009844:Oligosaccharides / Q000302:isolation & purification*
D009844:Oligosaccharides; D005643:Fucose