GC-MS and HPAEC
Purpose:
Qualitative and quantitative determination
of the monosaccharide composition of glycoconjugate, oligosaccharide, or
polysaccharide samples.
Methods Used:
Gas chromatography-mass spectrometry
(GC-MS) is usually used for samples originating from plants or microbes.
This analysis tests for the following carbohydrates:
arabinose
rhamnose
fucose
xylose
glucuronic acid
galacturonic acid
mannose
galactose
glucose
N-acetyl-mannosamine
N-acetyl-galactosamine
N-acetyl-glucosamine
High-pH anion-exchange chromatography
(HPAEC) is usually used for biomedical samples and samples originating
from animal cells. This analysis tests for the following carbohydrates:
fucose
glucose
mannose
galactose
N-acetyl-galactosamine
N-acetyl-glucosamine
HPAEC is also used for sialic acid determination and for oligosaccharide profiling.
General Criteria
for Sample Preparation:
Samples must be free of salts. If the
sample is a glycoprotein, it can be dialyzed and then lyophilized. If the
sample is an oligosaccharide, it can be desalted by size-exclusion chromatography
on a Bio-Gel P-2 or Sephadex G-10 column.
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