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Determination of the pKa of glucuronic acid and the carboxy groups of heparin by 13C-nuclear-magnetic-resonance spectroscopy.

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  • 1. Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City 52242.
      Authors
    • Wang HM 1
    (1 author)

The Biochemical Journal, 01 Sep 1991, 278 ( Pt 3):689-695
https://doi.org/10.1042/bj2780689 PMID: 1898357 PMCID: PMC1151401

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Abstract 

As part of our continuing studies on heparin, the present paper uses 13C-n.m.r. spectroscopy to examine the acidity of heparin's uronic acid carboxylate groups. Heparin contains three different uronic acids. In porcine mucosal heparin these account for approx. 91, 7 and 2 mol% of the total uronic acid residues. These are alpha-L-idopyranosyluronic acid 2-sulphate, beta-D-glucopyranosyluronic acid and alpha-L-idopyranosyluronic acid. The pKa values of their carboxylate groups were determined as 3.13 (using heparin), 2.79 (using heparin) and 3.0 (predicted by using model compounds) respectively. 18C-n.m.r. spectroscopy, performed at various pH values, provided a convenient method of simultaneously determining the pKa of multiple carboxylate groups, of similar acidity, within heparin D-Glucopyranosyluronic acid and heparin-derived di-, tetra- and hexa-saccharides were used as model compounds to determine pKa values of the different carboxy groups. The results suggested that molecular size had an effect on pKa. Unambiguous assignment of carboxy carbon resonances were accomplished through the use of two-dimensional n.m.r. spectroscopy. Finally, application of this method to the simplest model compound, D-glucopyranosyluronic acid, permitted the determination of the pKa of both its alpha- and beta-anomers.

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Biochem J. 1991 Sep 15; 278(Pt 3): 689–695.
PMCID: PMC1151401
PMID: 1898357

Determination of the pKa of glucuronic acid and the carboxy groups of heparin by 13C-nuclear-magnetic-resonance spectroscopy.

H M Wang, D Loganathan, and R J Linhardt
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Abstract

As part of our continuing studies on heparin, the present paper uses 13C-n.m.r. spectroscopy to examine the acidity of heparin's uronic acid carboxylate groups. Heparin contains three different uronic acids. In porcine mucosal heparin these account for approx. 91, 7 and 2 mol% of the total uronic acid residues. These are alpha-L-idopyranosyluronic acid 2-sulphate, beta-D-glucopyranosyluronic acid and alpha-L-idopyranosyluronic acid. The pKa values of their carboxylate groups were determined as 3.13 (using heparin), 2.79 (using heparin) and 3.0 (predicted by using model compounds) respectively. 18C-n.m.r. spectroscopy, performed at various pH values, provided a convenient method of simultaneously determining the pKa of multiple carboxylate groups, of similar acidity, within heparin D-Glucopyranosyluronic acid and heparin-derived di-, tetra- and hexa-saccharides were used as model compounds to determine pKa values of the different carboxy groups. The results suggested that molecular size had an effect on pKa. Unambiguous assignment of carboxy carbon resonances were accomplished through the use of two-dimensional n.m.r. spectroscopy. Finally, application of this method to the simplest model compound, D-glucopyranosyluronic acid, permitted the determination of the pKa of both its alpha- and beta-anomers.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
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