Fractionation of oligosaccharides containing N-acetyl amino sugars by reverse-phase high-pressure liquid chromatography

✍ Scribed by Karl Blumberg; Frederic Liniere; Lyubov Pustilnik; C.Allen Bush

• Publisher: Elsevier Science
• Year: 1982
• Tongue: English
• Weight: 425 KB
• Volume: 119
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(82)90605-4

✦ Synopsis

High-pressure liquid chromatography (hplc) of N-acetylglucosamine (GlcNAc) and galactosamine (GalNAc) containing carbohydrates was performed on several reverse-phase silica columns. Nanomolar level detection was accomplished using far uv-absorbance monitoring. Baseline separations of the (Y and fl anomers of GlcNAc, chitobiose, chitotriose, and chitotetraose were observed with water elution of the reverse-phase column. With the addition of up to 3% acetonitrile to the eluting solvent, similar resolution of chitin oligomers up to a chain length of seven was observed. Anomerization of the residue could be followed by isolation of either anomeric peak with subsequent rechromatography. Reduction of chitotriose with borohydride yielded a single sharp peak with a retention volume similar to that of the reducing trisaccharide. Semipreparative reverse-phase hplc allowed for the separation and identification by "C NMR of the GlcNAc-a-l-6

GlcNAc disaccharide from the p-I-6 isomer. Methyl glycosides of GalNAc and GlcNAc were shown to have retention times much longer than the free sugar.