Polysaccharides, paper chromatography

Shortly afterwards, Westphal, Liideritz, and their coworkers using the newly developed method of paper chromatography, found a new class of sugars in lipopolysaccharides (LPS) from Gram-negative bacteria, and identified them as 3,6-dideoxyhexoses. This work is summarized in Ref. 4. These discoveries initiated more-systematic investigations of hydrolyzates from bacterial polysaccharides, and a number of new monosaccharides were completely or partially identified. This development has been summarized by Ashwell and Hickman. ... 

Dermatan sulfate, also termed chondroitin sulfate B, a related glycosaminoglycan constituent of connective tissue, was known to be composed of galactosamine and a uronic acid, originally believed to be glucuronic acid but then claimed to be iduronic acid based largely on color reactions and paper chromatography. However, the d or L-enantiomer status of the latter monosaccharide was not clear. Jeanloz and Stoffyn unequivocally characterized the monosaccharide as L-iduronic acid by consecutive desulfation, reduction, and hydrolysis of the polysaccharide, followed by isolation of the crystalline 2,3,4-tri-0-acetyl-l,6-anhydro-/ -L-idopyranose, which was shown to be identical to an authentic specimen synthesized from 1,2-0-isopropylidene-/ -L-idofuranose.34... 

Acid hydrolysis of the basic hexasaccharide yielded the disaccharide 31. Its mobility in paper chromatography lay between those of the corresponding (1 — 3)-linked (from S10A) and /3-(l — 6)-linked (from S29) isomers, which is why it was assumed to be (1 — 4)-linked. The (1 — 2)-linked isomer was excluded, as the D-galactose residue that is part of 31 carries a D-galactopyranosyl group linked to 0-2 in the original polysaccharide, as will be discussed. 

Paper chromatography has been very widely used to analyze mixtures of the simple sugars and their phosphates, and to elucidate polysaccharide structures. The interest in these compounds is great. They have a widespread occurrence in complex natural mixtures which, before the advent of paper chromatography, presented exceedingly difficult, if not unsolv-able, problems in separation and purification. 

The monosaccharide composition of the products of the alkalii saponification of cellulose tosylates containing tosyloxy groups at Cg has been established on the basis of data obtained from a study of the composition of the products of acetylation, nitration, and periodate oxidation, and from quantitative paper chromatography of the products of complete hydrolysis of the mixed polysaccharides synthesized. The content of 3,6-anhydroglucose units in the 3,6-anhydro derivatives of cellulose was found to be 50-70 mol.-% (10-13). 

No structural work has been done on the polysaccharide preparations. In the hydrolyzates, the sugar components have been estimated quantitatively by paper chromatography or other methods (see Table II). It would be premature to try to interpret the ratios found between individual sugars or classes of sugars, or to discern a pattern in the variations between different soils, because the preparations were heterogeneous and repre.sented only a small part of the total polysaccharides of the soil. 

The preceding examples elegantly demonstrate the point at which zone electrophoresis may be used to advantage in structural determinations on polysaccharides, namely, in further resolving fractions isolated by carbon-Celite column chromatography or paper chromatography, or both. 

The acid is reduced by sugars and by ascorbic acid to a dye, and it can be used for microdetermination of sugar components of polysaccharide hydrolyzates after separation by paper chromatography. ... 

---