Glycosaminoglycans

Purpose

To determine the positions of linkage of glycosyl residues in an oligosaccharide, polysaccharide, or glycoprotein. The linkage of the residues, including percentage of each is provided with this analysis.

Linkage analysis of neutral, uronic acids or amino-residues (methylation analysis) – The polysaccharides or oligosaccharides are derivatized prior to GC-MS analysis. This procedure uses either the Hakomori, or NaOH base, for the permethylation procedure.

Linkage analysis of Kdo– Kdo (2-keto-3-deoxy-octulosonic acid) is readily analyzed by a modified linkage procedure.

Methods used

For glycosyl linkage analysis, the sample is permethylated, depolymerized, reduced, and acetylated; and the resultant partially methylated alditol acetates (PMAAs) are analyzed by gas chromatography-mass spectrometry (GC-MS).

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Amount required for sample analysis

A minimum of 100-200 micrograms of carbohydrate is required, more if available. Your sample must be pure, free of salt and detergent.

Unusual Residues

The CCRC has over 30 years of experience with linkage analysis and has accumulated a vast library of spectra on unusual monosaccharides and their linkages. We have the experience and expertise to determine the glycosyl linkage of any non-routine residue. We can also develop a specific methylation procedure for linkage analysis of your particular glycan if needed.  Glycosyl composition data are usually required prior to glycosyl linkage analysis.