Analysis of glycosaminoglycan-derived oligosaccharides using reversed-phase ion-pairing and ion-exchange chromatography with suppressed conductivity detection

Analytical Biochemistry
R J LinhardtS R Carter

Abstract

Oligosaccharides prepared from glycosaminoglycans (GAGs) including heparin, heparan sulfate, chondroitin sulfates, dermatan sulfate, and keratan sulfate were analyzed using reverse-phase ion-pairing HPLC and ion-exchange HPLC with suppressed conductivity detection. The results were compared with those obtained by strong anion-exchange HPLC using uv detection. These oligosaccharides were first prepared by enzymatically depolymerizing the GAGs with enzymes including heparin lyase (EC 4.2.2.7), heparan sulfate lyase (EC 4.2.2.8), chondroitin ABC lyase (EC 4.2.2.4), and keratan sulfate hydrolase (EC 3.2.1.103). Analysis was then performed without derivitization under isocratic conditions with a limit of sensitivity in the picomole range. Preliminary studies suggest that this approach may be particularly useful in examining oligosaccharides having no uv chromophore such as those prepared from keratan sulfate.

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Citations

Sep 21, 2010·Analytical and Bioanalytical Chemistry·Bo YangRobert J Linhardt
Mar 7, 2012·Applied Microbiology and Biotechnology·C K M TripathiVikas Mishra
Jan 12, 1996·Journal of Chromatography. a·R J Linhardt, A Pervin
Jan 10, 1997·Journal of Chromatography. B, Biomedical Sciences and Applications·P Y Johnson, D A Blake
Nov 2, 2016·Analytical Chemistry·Rebecca L MillerJeremy E Turnbull
Mar 13, 1990·Biochemistry·R J LinhardtJ T Gallagher
Nov 1, 1996·Biomedical Chromatography : BMC·M J Davies, E F Hounsell
Jan 1, 2012·Metabolites·Tania M Puvirajesinghe, Jeremy E Turnbull
Jul 27, 2007·Proteomics·Bernd GesslbauerAndreas J Kungl
Jan 23, 2016·Journal of Separation Science·Jennifer M Fasciano, Neil D Danielson
Nov 1, 1990·Analytical Biochemistry·D A Blake, N V McLean

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