Identification and fractionation of human milk oligosaccharides by proton-nuclear magnetic resonance spectroscopy and reverse-phase high-performance liquid chromatography

✍ Scribed by Virendra K. Dua; C.Allen Bush

• Book ID: 102986903
• Publisher: Elsevier Science
• Year: 1983
• Tongue: English
• Weight: 502 KB
• Volume: 133
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(83)90215-4

✦ Synopsis

It has been shown that the retention times of oligosaccharides containing N-acetyl amino sugars in reverse-phase high-performance liquid chromatography are sensitive both to chain length and to stereochemical differences. Data showing that oligosaccharides of human milk can be fractionated according to stereochemistry on C-18 columns using water as eluant, is presented. Detection at the nanomolar level is possible using ultraviolet absorption at 190 nm as a result of the acetamido group. The separation of linkage isomers by fractionation of two tetrasaccharides of identical carbohydrate composition, lacto-N-tetraose and lacto-iV-neotetraose, and by the sep aration of two isomeric pentasaccharides, lacto-N-fucopentaose I and II, is shown. In some cases it was possible to preparatively fractionate pure oligosaccharides from complex mixtures. The identity and purity of the oligosacchatides was determined by proton-nuclear magnetic resonances spectroscopy at 300 MHz using the method of Vliegenthart and co-workers (B. Foumet, G.