IMPROVED PROTOCOL FOR THE FORMATION OF N-(P-NITROBENZYLOXY)AMINOALDITOL DERIVATIVES OF OLIGOSACCHARIDES

摘要

An improved procedure has been developed for the rapid derivatization of oligosaccharides with W-detectable p-nitrobenzylhydroxylamine (PNB). The improved conditions used result in quantitative derivatization of neutral oligosaccharides. Sialylated oligosaccharides can also be quantitatively PNB-derivatized without detectable desialylation. Of the oligosaccharides tested, only the derivatization of oligogalactosyluronic acids was incomplete (yield similar to 70%). PNB-derivatization of tamarind seed xyloglucan oligosacccharides results in products with improved chromatographic properties during HPAEC, These PNB derivatives were also subjected to hydrophilic interaction chromatography (HILIC) and analyzed by on-line LC-MS. On-line LC-MS is readily usable with HILIC, as this chromatographic technique does not require salt-containing solvents. Approximately 10 pmol of a PNB-derivatized oligosaccharide can be identified and quantitated utilizing this method. [References: 11]