Separation of fucosylated oligosaccharides using high-pH anion-exchange chromatography with pulsed-amperometric detection

✍ Scribed by Mark R. Hardy; R.Reid Townsend

• Publisher: Elsevier Science
• Year: 1989
• Tongue: English
• Weight: 450 KB
• Volume: 188
• Category: Article
• ISSN: 0008-6215
• DOI: 10.1016/0008-6215(89)84052-2

✦ Synopsis

Fucosylated oligosaccharides of the /3Gal(l+4)GlcNAc-, PGal(l+4)Glc-, and /3Gal(l+3)GlcNAc+eries were chromatographed on a high-performance anion-exchange pellicular resin under alkaline conditions (pH ~13). Fucosylation of either lactose, lactosamine (Type II chains), or lacto-N-biose (Type I chains) oligosaccharides markedly decreased the retention time (l&38 min) of the nonfucosylated form. The magnitude of the reduction was related to whether fucose replaced Gal [cuFuc(l+3)+GlcNAc],

whether fucose was a-(1+2)-linked> Gal at the end of a chain, or whether fucose was linked in a subterminal position [a(1+3) or a(l+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 anionexchange chromatography can effectively separate 1+2, 1+3, and 1+4 fucose positional isomers in a single chromatographic step.