PubMed:18368478 / 5-16
Determination of iduronic acid and glucuronic acid in sulfated chondroitin/dermatan hybrid chains by (1)H-nuclear magnetic resonance spectroscopy.
The relative proportion of L: -iduronic acid (IdoA) and D: -glucuronic acid (GlcA) is of great importance for the structure-function relationship of chondroitin sulfate (CS)/dermatan sulfate (DS). However, determination of the isotypes of uronic acid residues in CS/DS is still a challenge, due to the instability of free uronic acid released by chemical degradation and its conversion to unsaturated uronic acid by digestion with bacterial eliminase. (1)H-Nuclear magnetic resonance (NMR) spectroscopy is a promising tool with which to address this issue, but the traditional method based on the assignment of the ring proton signals of IdoA and GlcA residues still has drawbacks such as the serious overlap of signals in the (1)H-NMR spectrum of CS/DS polysaccharides. We found that the proton signals of the N-acetyl group of N-acetyl-D: -galactosamines in CS and DS could be clearly distinguished and accurately integrated in the one-dimensional (1D) (1)H-NMR spectrum. Based on this finding, here we report a novel, sensitive, and nondestructive 1D (1)H-NMR-based method to determine the proportion of IdoA and GlcA residues in CS/DS hybrid chains.
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