Practical Protein Chemistry - A. Darbre 1989

Analytical Methods
Glycoproteins
Determination of alditols by thin-layer and gas-liquid chromatography

Reduced and permethylated Oligosaccharides (1–2 mg, containing up to 15 monosaccharide residues) were separated on TLC silica gel 60 plates (250 µm, Merck) without prior activation, using a benzene – methanol (16:1 or 10:1) solvent system. The sample was applied as a band, and Chromatography was performed twice in chambers whose inner walls were lined with filter paper soaked in the Solvents (to ensure complete saturation). Detection was carried out by spraying with a 0.5% orcinol solution in 2 M sulfuric acid. The detection limit is 0.5 µg of sugar.

The decisive factors in the Separation of Disaccharides are THE POSITION OF the glycosidic bond, the configuration of the anomer, and The Nature of the sugar.

Gas-Liquid Chromatography. Fused-silica capillary Column (12 m × 0.25 mm), with walls coated with a chemically bonded DB-1 phase (0.1 µm film thickness, J. and W. Scientific); carrier gas: helium, linear velocity 0.37 cm/s; Temperature program: heating from 150 °C to 330 °C at a rate of 5 °C/min. Sensitivity: 30 ng for sophorose (GIc ß1-2Glе).



Last update: 06/08/2026

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