Analysis of procainamide-derivatised heparan sulphate disaccharides in biological samples using hydrophilic interaction liquid chromatography mass spectrometry

Analytical and Bioanalytical Chemistry
Imeobong U AntiaAjit J Shah

Abstract

Glycosaminoglycans (GAGs) are a family of linear heteropolysaccharides made up of repeating disaccharide units that are found on the surface and extracellular matrix of animal cells. They are known to play a critical role in a wide range of cellular processes including proliferation, differentiation and invasion. To elucidate the mechanism of action of these molecules, it is essential to quantify their disaccharide composition. Analytical methods that have been reported involve either chemical or enzymatic depolymerisation of GAGs followed by separation of non-derivatised (native) or derivatised disaccharide subunits and detection by either UV/fluorescence or MS. However, the measurement of these disaccharides is challenging due to their hydrophilic and labile nature. Here we report a pre-column LC-MS method for the quantification of GAG disaccharide subunits. Heparan sulphate (HS) was extracted from cell lines using a combination of molecular weight cutoff and anion exchange spin filters and digested using a mixture of heparinases I, II and III. The resulting subunits were derivatised with procainamide, separated using hydrophilic interaction liquid chromatography and detected using electrospray ionisation operated in positive...Continue Reading

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Citations

Jul 7, 2020·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Yuefan SongRobert J Linhardt
Sep 18, 2020·Molecular & Cellular Proteomics : MCP·Lauren E PepiI Jonathan Amster
Aug 7, 2021·Analytical Chemistry·Zhangjie WangJian Liu
Oct 15, 2020·Carbohydrate Polymers·Matěj ŠimekVladimír Havlíček
Oct 6, 2021·Mass Spectrometry Reviews·Andreas ZappeKevin Pagel
Oct 16, 2021·Analytical and Bioanalytical Chemistry·Salomé PoyerRégis Daniel

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