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Quantitative Analysis of Neutral and Acidic Sugars in Whole Bacterial Cell Hydrolysates Using High-performance Anion-exchange Liquid Chromatography–electrospray Ionization Tandem Mass Spectrometry by D.S Wunschel; K.F Fox; A Fox; M.L Nagpal; K Kim; G.C Stewart; M Shahgholi is a Chemistry article available to read on EtoBox.

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A procedure for analysis of a mixture of neutral and acidic sugars in bacterial whole cell hydrolysates using high-performance anion-exchange liquid chromatography-electrospray ionization tandem mass spectrometry (HPAEC-ESI-MS-MS) is described. Certain bacteria (including bacilli), grown under phosphate-limited conditions, switch from producing a teichoic acid (containing ribitol) to a teichuronic acid (characterized by glucuronic acid content). Bacterial cells were hydrolyzed with sulfuric acid to release sugar monomers. The solution was neutralized by extraction with an organic base. Hydrophobic and cationic contaminants (including amino acids) were removed using C18 and SCX columns, respectively. HPAEC is well established as a high-resolution chromatographic technique, in conjunction with a pulsed amperometric detector. Alternatively, for more selective detection, sugars (as M-H- ions) were monitored using ESI-MS. In HPAEC, the mobile phase contains sodium hydroxide and sodium acetate, which are necessary for chromatographic separation of mixtures of neutral and acidic sugars. Elimination of this high ionic content prior to entry into the ESI ion source is vital to avoid comprom

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Author

D.S Wunschel; K.F Fox; A Fox; M.L Nagpal; K Kim; G.C Stewart; M Shahgholi

Publisher

Elsevier Science; Elsevier ; Elsevier BV (ISSN 1873-3778)

Published

1997

Language

EN

Field

Chemistry (Physical Sciences)