Improved protocol for the formation of N-(p-nitrobenzyloxy)aminoalditol derivatives of oligosaccharides.
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.
Pauly
M
M
Department of Biochemistry and Molecular Biology, University of Georgia, Athens 30602-4712, USA.
York
W S
WS
Guillen
R
R
Albersheim
P
P
Darvill
A G
AG
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
D006898:Hydroxylamines; D009844:Oligosaccharides; D013402:Sugar Alcohols; C037705:4-nitrobenzyloxyamine