Journal
CARBOHYDRATE RESEARCH
Volume 342, Issue 5, Pages 724-735Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.carres.2006.12.007
Keywords
MALDI-TOF/TOF; arabinoxylan; isomers; high-energy CID; normal phase HPLC
Funding
- Biotechnology and Biological Sciences Research Council [D19624] Funding Source: researchfish
- Biotechnology and Biological Sciences Research Council [D19624] Funding Source: Medline
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Normal phase-high performance liquid chromatography (NP-HPLC) coupled to matrix-assisted laser desorption/ionization-time-of-flight/time-of-flight (MALDI-TOF/TOF) tandem mass spectrometry is evaluated for the detailed structural characterization of various isomers of arabinoxylan (AX) oligosaccharides produced from endo-beta-(1 -> 4)-xylanase (endoxylanase) digestion of wheat AX. The fragmentation characteristics of these oligosaccharides upon MALDI-TOF/TOF high-energy collision induced dissociation (CID) were investigated using purified AX oligosaccharide standards labeled at the reducing end with 2-aminobenzoic acid (2-AA). A variety of cross-ring cleavages and 'elimination' ions in the fragment ion spectra provided extensive structural information, including Araf substitution patterns along the xylan backbone and comprehensive linkage assignment. The off-line coupling of this MALDI-CID technique to capillary normal phase HPLC enabled the separation and identification of isomeric oligosaccharides (DP 4-8) produced by endoxylanase digestion of AX. Furthermore, this technique was used to characterize structurally different isomeric AX oligosaccharides produced by endoxylanase enzymes with different substrate specificities. (c) 2006 Elsevier Ltd. All rights reserved.
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