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

• 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
• ISSN: 0021-9673; 1873-3778
• DOI: 10.1016/j.chroma.2008.01.065

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).