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HS and HP chains vary in the sulfation pattern and the degree of C\u20105 epimerization of <jats:sc>d<\/jats:sc>\u2010glucuronic acid to <jats:sc>l<\/jats:sc>\u2010iduronic acid. These modifications are not uniformly distributed within the chain, providing functional oligomeric domains interacting specifically with various effective proteins. The utilization of specific lyases and chemical depolymerization are the commonest procedures used for structural analysis. Di\u2010 and oligosaccharide composition of HS can be accurately and sensitively determined by HPLC, CE and MS. Ultraviolet detection is satisfactory enough for unsaturated saccharides and pre\u2010column derivatization with fluorophores and detection with laser\u2010induced fluorescence results in even higher sensitivity. Solid\u2010phase assays can also be used for monitoring interactions with other molecules. In this article the biological significance of HS and HP in health and disease as well as the portfolio of analytical methods that may help to a deeper understanding of their roles in various pathological processes is presented. Such methodologies are of crucial importance for disease diagnosis and the design of novel synthetic sugar\u2010based drugs. 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