How Are the Repeating Units of Polysaccharides Determined through Analytical Approaches?
During the structural analysis of polysaccharides, the repeating units can be elucidated through the following sequential steps:
Separation and Purification
Chromatographic techniques, such as gel permeation chromatography (GPC), are employed to separate and purify polysaccharide samples, ensuring the isolation of fractions with consistent molecular weight distributions.
Hydrolysis
The purified polysaccharides are subjected to acid hydrolysis, cleaving the glycosidic bonds to yield monosaccharides or oligosaccharide fragments. This step facilitates subsequent structural elucidation.
Monosaccharide Composition Analysis
The hydrolyzed products are analyzed using high-performance liquid chromatography (HPLC) or gas chromatography (GC) to identify the types and relative abundance of monosaccharide residues present in the polysaccharide.
Linkage Analysis
Glycosidic linkages between monosaccharide units are characterized using mass spectrometry techniques (e.g., MALDI-TOF MS) in combination with nuclear magnetic resonance (NMR) spectroscopy, enabling the determination of linkage positions and configurations.
Sequence Analysis
By integrating NMR spectral data with mass spectrometric analysis, the sequence of monosaccharides within the repeating unit can be established, leading to the structural identification of the polysaccharide's repeating motif.
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