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Article Dans Une Revue Separations Année : 2020

Analysis of Chondroitin/Dermatan Sulphate Disaccharides Using High-Performance Liquid Chromatography

Ivan Mikšík
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Šárka Kubinová
Karel Výborný
Ameneh Tatar
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Vladimír Král
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Kamil Záruba
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David Sýkora

Résumé

Chondroitin sulphates belong to a group of naturally occurring glycosaminoglycans and play a role in many physiological processes including ageing and the effects of various diseases. Research into chondroitin sulphates has found that the most important analytes are 4- and 6-sulphated disaccharides. We developed an HPLC method for the separation and quantification of underivatized chondroitin/dermatan sulphates—unsaturated disaccharides (4- and 6-sulphated disaccharides). This method is based on the separation of disaccharides by amido as well as amino columns under acidic conditions. These columns enabled the successful separation of 4- and 6-sulphated disaccharides using 50 (amido column) and 25 mmol/L (amino column) phosphate buffer, pH 4.25 (detection at 230 nm), at retention times of less than 10 min. The limit of quantification was 0.5 μg/mL. The applicability of this method was demonstrated through analysis of unsaturated disaccharides produced from the enzymatic digestion of chondroitin/dermatan sulphates of the solubilized extracellular matrix produced from porcine urinary bladder and human umbilical cord.
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Dates et versions

hal-04313979 , version 1 (29-11-2023)

Identifiants

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Ivan Mikšík, Šárka Kubinová, Marine Morvan, Karel Výborný, Ameneh Tatar, et al.. Analysis of Chondroitin/Dermatan Sulphate Disaccharides Using High-Performance Liquid Chromatography. Separations, 2020, 7 (3), pp.49. ⟨10.3390/separations7030049⟩. ⟨hal-04313979⟩
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