Activated sludge extracellular polymers (ECP) were extracted either by sonication or a combination of sonication and cation exchange resin treatment (CER). The chemical composition of the aqueous extract was investigated by chromatographic analysis of amino acids and sugars after hydrolysis. Up to 70 to 80% of the total organic carbon (TOC) of ECP was characterized. Proteins were found to be the major constituent of ECP, which was confirmed by pyrolysis/GC/MS analysis. Sugar and protein analysis led to complementary information both on the origin of extracellular material and on sludge floc structure. The monosaccharide composition in ECP and sludge allowed the proposal of different origins for extracellular polysaccharides. The predominance of proteins in ECP underlined their key role in the floc structure, and led to hypotheses on their origin. Proteins were better extracted than sugars when the CER extraction was combined with sonication. This supposed that proteins are more involved than sugars in electrostatic bonds with multivalent cations. Electrostatic bonds were found to be uniformly distributed in the floc and closely combined with hydrophobic bonds. About 25% of ECP amino acids were negatively charged and 24% exhibited hydrophobic properties, highlighting the specific role of proteins in the floc structure.

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