Oligosaccharide Compositional Analysis of Heparin and Low Molecular Weight Heparins by Capillary Electrophoresis

Authors
Desai, U.R.
Wang, H.M.
Ampofo, S.A.
Linhardt, Robert J.
ORCID
https://orcid.org/0000-0003-2219-5833
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Other Contributors
Issue Date
1993
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
Degree
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Full Citation
Oligosaccharide Compositional Analysis of Heparin and Low Molecular Weight Heparins by Capillary Electrophoresis, U.R. Desai, H.M. Wang, S.A. Ampofo, R.J. Linhardt, Analytical Biochemistry, 213, 120-127 (1993).
Abstract
The application of capillary electrophoresis to total compositional analysis of heparin and low-molecular-weight heparin samples has been studied. Optimum resolution of 17 defined oligosaccharides was obtained with the buffer system composed of 10 mM sodium borate and 50 mM sodium dodecyl sulfate at pH 8.81 and at a constant voltage of 20 kV. The ratio of oligosaccharide charge to the number of saccharide residues correlated with the migration time. For oligosaccharides having the same charge to saccharide ratio, the larger of the oligosaccharides eluted earlier. A hexasaccharide having a 3-O-sulfated glucosamine residue at the reducing end and arising from heparin's antithrombin III binding site, migrated in an unusual fashion. The limit of oligosaccharide detection was from 600 fmol to 1 pmol. Quantitative analysis could conveniently be performed on 10 pmol of an oligosaccharide sample. Oligosaccharide composition using capillary electrophoresis was obtained by nearly complete depolymerization of heparins with a mixture of heparin lyase I, II, and III. The analysis resulted in 95% mass balance for both heparin and low-molecular-weight heparin. Capillary electropherograms of heparin and different low-molecular-weight heparins depolymerized with heparin lyase I alone showed a high level of structural heterogeneity in the products formed. The oligosaccharide maps thus obtained might find use in fingerprinting the heparin and low-molecular-weight samples.
Description
Analytical Biochemistry, 213, 120-127
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Department
The Linhardt Research Labs.
The Shirley Ann Jackson, Ph.D. Center for Biotechnology and Interdisciplinary Studies (CBIS)
Publisher
Relationships
The Linhardt Research Labs Online Collection
Rensselaer Polytechnic Institute, Troy, NY
https://harc.rpi.edu/
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