Paper chromatography of acidic carbohydrates

CATIONIC detergents, such as cetyltrimethylammonium bromide (‘Cetavlon’) and cetylpyridinium chloride, have been used extensively in recent years for the fractionation of polysaccharide mixtures1–4. The quaternary ammonium cations present in these detergents form salts with acidic polysaccharides which often have solubility properties quite different from those of the usual metallic salts; for example1, cetyltrimethylammonium chondroitin sulphate is soluble in aliphatic alcohols up to n-pentanol and is insoluble in water. Many purification procedures have utilized the water-insolubility of these quaternary ammonium polysaccharide salts, but surprisingly, no serious attempt seems to have been made to exploit their solubility in organic solvents. This communication describes some preliminary experiments directed towards developing convenient methods for the separation of acidic carbohydrates, making use of the solubility of their cetylpyridinium salts in organic solvents. Current interest in these laboratories in the purification and identification of sugar sulphates, suggested that it would be useful if this property could be utilized in a method for the paper chromatography of these substances. It has been found that when a small amount of detergent is incorporated into the mobile (organic) phase of certain solvent systems for paper chromatography, the R F values for sulphated derivatives are increased by a factor of about ten, and some useful separations can be achieved. Two such systems are