title: Improved protocol for the formation of N-(p-nitrobenzyloxy)aminoalditol derivatives of oligosaccharides.
abstract: An improved procedure has been developed for the rapid derivatization of oligosaccharides with UV-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 approximately 70%). PNB-derivatization of tamarind seed xyloglucan oligosaccharides 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.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Georgia, Athens 30602-4712, USA.
authors.forename: M
authors.initials: M
authors.lastname: Pauly
authors.forename: W S
authors.initials: WS
authors.lastname: York
authors.forename: R
authors.initials: R
authors.lastname: Guillen
authors.forename: P
authors.initials: P
authors.lastname: Albersheim
authors.forename: A G
authors.initials: AG
authors.lastname: Darvill
mesh_terms: D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D006898:Hydroxylamines; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000138:chemical synthesis / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet; D013402:Sugar Alcohols
chemical_list: D006898:Hydroxylamines; D009844:Oligosaccharides; D013402:Sugar Alcohols; C037705:4-nitrobenzyloxyamine