Chitosan is a widely used linear biopolymer composed mainly of glucosamine and to a lesser extent of N-acetylglucosamine units. Many biological activities of chitosan are attributed to its shorter oligomeric chains, which consist of chitosan prepared either by enzyme activity (lysozyme, bacterial chitinase) or chemically by acid-catalyzed hydrolysis (e.g. in the stomach). However, these processes always result in a mixture of shorter chitooligosaccharides with varying degrees of acetylation whereas for relevant results of biological studies it is necessary to work with a precisely defined material. In this review, we provide an overview and comparison of analytical methods leading to the determination of the degree of polymerization (DP), the degree of acetylation (DA), the fraction of acetylation (FA) and the acetylation patterns (PA) of chitooligosaccharide chains and of the current state of knowledge on chitooligosaccharide separation. This review aims to present the most promising routes to well-defined low molecular weight chitosan with low dispersity.

Laboratory of Biotransformation Institute of Microbiology of the Czech Academy of Sciences Vídeňská 1083 CZ 14200 Prague Czech Republic

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