**1. Introduction**

304 The Complex World of Polysaccharides

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Polysaccharides play important biological roles in all living beings. They act as membrane and cell wall constituents, afford physical and chemical protection from the environment, ensure nutrition storage or compose the antigens that increase or repress defenses during infection processes, protein folding, molecular recognition, cell adhesion, etc. [1,2]. With regard to their major biological roles it appears a priority to understand their structure/activity relationship. Unfortunately, even though their relevance has been clearly demonstrated, the structural analysis of such compounds remains difficult. Actually, even though they are often highly soluble in water, their polarity due to the presence for instance of hydroxyls, carboxyls and sulfates, makes them quite difficult to handle. These polar groups are involved in intra and inter-molecular hydrogen bonding, ionic interactions and for lipopolysaccharides (LPS) hydrophobic interactions. Such molecular interactions are sources of solubility problems and can lead to the formation of aggregates, making their analysis more difficult (enormous apparent size, broadening of NMR signals, ionization difficulties, and so on).

In the past, analysis of polysaccharides was mostly achieved through Gas Chromatography coupled to Mass Spectrometry (GC-MS) of the PS hydrolysates. This technique remains widely used, due to its simplicity, but the development of high field Nuclear Magnetic

© 2012 Poinsot et al., licensee InTech. This is an open access chapter distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. © 2012 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Resonance (NMR) and cryoprobes have opened new avenues. Unfortunately, NMR presents major drawbacks such as sample purity, sample complexity and concentration requirements. However, in the last decade, the development of highly sensitive, highthroughput Mass Spectrometers (MS) and software able to make global approaches ("omics") on very complex mixtures, have induced a new interest for the MS analyses (especially for the study of protein glycosylation). This time, the instruments mostly use soft ionization techniques associated to powerful MS2 or n capacity analyzers experiments on electrospray ionization coupled to tandem mass spectrometers (ESI-MS/MS) or matrix assisted laser desorption and ionization coupled to a time of flight analyser (MALDI-ToF) [3]. Finally, the recent appearance of ToF and FTICR (Fourier transform ion cyclotron resonance) analyzers on the market allowed access to high resolution measurements enabling the determination of the exact molecular masses (with ToF error of less than 5 ppm and FTICR less than 0.5 ppm). Exact masses give access to the elementary composition of the molecule that is extremely precious when the analyte is present in a complex biological matrix at low concentrations precluding study by MS/MS.

Beside the evolution of structural analysis, separations techniques have also been enhanced especially by using higher resolution techniques (narrow bore columns, high pressure tolerant or acido-basic tolerant chromatographic phases). This progress has made it possible to couple chromatographic and MS systems on-line.

In this chapter, we will describe the classical MS couplings like GC-MS or HPLC-MS, but also focus on unusual but useful polysaccharide analysis systems like capillary electrophoresis coupled with mass spectrometry (CE-MS) or high performance anion exchange chromatography coupled with mass spectrometry (HPAEC-MS) that we have developed. Finally, we will present the practical uses of these techniques in a fundamental application: the analysis of the exo-, lipo- and capsular polysaccharides of *Sinorhizobium meliloti.* 
