Analysis of glycosaminoglycan-derived oligosaccharides using reversed-phase ion-pairing and ion-exchange chromatography with suppressed conductivity detection.
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.
Linhardt
R J
RJ
Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City 52242.
Gu
K N
KN
Loganathan
D
D
Carter
S R
SR
D002809:Chondroitin Sulfates / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003871:Dermatan Sulfate / Q000032:analysis; D004553:Electric Conductivity; D006025:Glycosaminoglycans / Q000032:analysis*; D006493:Heparin / Q000032:analysis; D007632:Keratan Sulfate / Q000032:analysis; D008970:Molecular Weight; D009844:Oligosaccharides / Q000032:analysis* / Q000302:isolation & purification; D013056:Spectrophotometry, Ultraviolet
D006025:Glycosaminoglycans; D009844:Oligosaccharides; D003871:Dermatan Sulfate; D006493:Heparin; D002809:Chondroitin Sulfates; D007632:Keratan Sulfate