Linear ion trap MSn of enzymatically synthesized 13C-labeled fructans revealing differentiating fragmentation patterns of β (1-2) and β(1-6) fructans and providing a tool for oligosaccharide identification in complex mixtures
Publication: Research - peer-review › Journal article – Annual report year: 2012
Fructans are polymeric carbohydrates, which play important roles as plant reserve carbohydrates and stress protectants, and are beneficial for human health and animal production. Fructans are formed by the addition of Î-d-fructofuranosyl units to sucrose, leading to very complex mixtures of 1-kestose based inulins, 6-kestose linked levans, and 6G-kestose derived neoseries inulins and levans in cool season grasses such as Lolium perenne. The identification of isomeric fructan oligomers in chromatographic analysis of crude plant extracts is often hampered by the lack of authentic standards, and unambiguous peak assignment usually requires time-consuming analyses of purified fructan oligomers. We have developed a LC-MSn method for the separation and detection of fructan isomers and present here evidence for specific MSn fragmentation patterns associated with β1-2 (inulins) and β2-6 (levans) fructans. LC-MSn analysis of 13C labeled fructan oligomers produced by L. perenne fructosyltransferases expressed in yeast has enabled us to account for the observed fragmentation patterns in terms of preferential cleavage of the glycosidic bond between O- and fructose C2 in both inulins and levans and to differentiate reducing-end from nonreducing end cross ring cleavages in levans. We propose that higher order MS fragmentation patterns can be used to distinguish between the two major classes of fructan, i.e., inulins and levans, without the need for authentic standards. © 2011 American Chemical Society.
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