Variations of pH as an additional tool in the analysis of crowded NMR spectra of fucosylated chondroitin sulfates

• NMR analysis of three fucosylated chondroitin sulfates was performed at different pH.
• The signals of glucuronic acid residues were the most sensitive to pH changes.
• The H-1 signals of GlcA and GalNAc were resolved at pH 7.2.
• The H-1 signals of GlcA3S and GalNAc were resolved at pH 2.0.
• Combination of data obtained at different pH helps in NMR spectra assignment of FCS.

The influence of pH variation on chemical shift values in NMR spectra of fucosylated chondroitin sulfates was studied using polysaccharides isolated from three sea cucumber species Apostichopus japonicus, Actinopyga mauritiana and Cucumaria japonica. The signals of glucuronic acid residues were found to be the most sensitive to pH changes in comparison to the chemical shifts of the sulfated galactosamine and fucosyl units, most of which were altered insignificantly. It was shown that in the presence of imidazole-HCl buffer (pH 7.2) NMR spectra of the polysaccharides from A. japonicus and A. mauritiana were sufficiently resolved, whereas under acidic conditions their 1H NMR spectra were complicated by overlapping of H-1 signals of GlcA and GalNAc. In the case of polysaccharide from C. japonica bearing 3-O-fucosylated and 3-O-sulfated glucuronic acid residues in the backbone, acidification of the medium led to separation of H-1 signals of GlcA3S and GalNAc. Therefore, the combination of data obtained at different pH values may be useful for interpretation of overcrowded spectra of fucosylated chondroitin sulfates.

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