Rapid molecular mass and structural determination of plant cell wall-derived oligosaccharides using off-line high-performance anion-exchange chromatography/mass spectrometry.
A method has been developed for the rapid molecular mass determination and structural elucidation of mixtures of oligosaccharides derived from plant cell walls. The oligosaccharides were fractionated using gel permeation chromatography and 'analytical' high-performance anion-exchange chromatography (HPAEC), neutralized, dried and the mixtures of eluent salt and oligosaccharides were per-O-acetylated directly. The derivatized oligosaccharides were isolated by dissolution in dichloromethane and the salts were removed by aqueous partitioning. The per-O-acetylated oligosaccharides were analysed using electrospray (ES) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MS). Exploiting the fact that acid-catalysed per-O-acetylation of oligosaccharides can be achieved even under the extremely salty conditions that are found in post-column neutralized HPAEC fractions, and combining this derivatization step with off-line ESMS, allow rapid screening for molecular mass and thus yield information on the composition of the various oligosaccharides in these complex mixtures. Subsequent per-O-methylation of the per-O-acetylated, salt-free fractions and collision-induced dissociation tandem mass spectrometric analysis was used for additional sequence and branching determination of the oligosaccharides.
Brüll
L
L
Bijvoet Center for Biomolecular Research, Department of Mass Spectrometry, Faculty of Chemistry, Utrecht University, The Netherlands.
Huisman
M
M
Schols
H
H
Voragen
F
F
Critchley
G
G
Thomas-Oates
J
J
Haverkamp
J
J
D002240:Carbohydrate Sequence; D002473:Cell Wall / Q000737:chemistry; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry*; D059828:Plant Cells; D010944:Plants / Q000737:chemistry*; D013025:Glycine max / Q000737:chemistry / Q000166:cytology; D016339:Spectrometry, Mass, Fast Atom Bombardment
D009844:Oligosaccharides