Polysaccharides analysis of sinorhizobial capside by on-line anion exchange chromatography with pulsed amperometric detection and mass spectrometry coupling

High performance anion exchange chromatography (HPAEC)-pulsed amperometric detection (PAD) is a performing technique for carbohydrate analysis, due to the selectivity and sensitivity of the detection. The identification occurs through retention times. In absence of standards, structural characteriza...

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Bibliographic Details
Published in:Journal of Chromatography A 2008-03, Vol.1185 (2), p.241-250
Main Authors: Chataigné, G., Couderc, F., Poinsot, V.
Format: Article
Language:English
Subjects:
Analytical chemistry
Bacterial Capsules - analysis
Bacterial Capsules - isolation & purification
Bacteriology
Biological and medical sciences
Capsular polysaccharide
capsular polysaccharides
Chemical Sciences
Chromatography, Ion Exchange - methods
electrochemistry
Fundamental and applied biological sciences. Psychology
glucans
Glycerophosphorylated glucan
high performance anion exchange chromatography
HPAEC-PAD
HPAEC-PAD–MS
ion exchange chromatography
mass spectrometry
Microbiology
Morphology, structure, chemical composition
on-line anion exchange chromatography
polysaccharides
pulsed amperometric detection
Sensitivity and Specificity
Sinorhizobium
Sinorhizobium - chemistry
Sinorhizobium fredii - chemistry
Sinorhizobium meliloti
Sinorhizobium meliloti - chemistry
Spectrometry, Mass, Electrospray Ionization - methods
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Description
Summary:High performance anion exchange chromatography (HPAEC)-pulsed amperometric detection (PAD) is a performing technique for carbohydrate analysis, due to the selectivity and sensitivity of the detection. The identification occurs through retention times. In absence of standards, structural characterization of complex polysaccharides requests the coupling of HPAEC-PAD with electrospray ionization (ESI)-MS. This is a technological challenge, due to the non-volatility and high conductance of the eluents. Therefore, a desalting device has been installed on-line between the PAD and the MS. On-line HPAEC–MS has only been rarely described. We report here successful analysis of biological acidic oligosaccharides, allowing for the first time to demonstrate that membrane anchored 3-deoxy- d-manno-2 octulosonic acid (Kdo) homopolymers are consensus sinorhizobial capsular polysaccharide (KPS).
ISSN:0021-9673
1873-3778
DOI:10.1016/j.chroma.2008.01.065