The partition of glycosaminoglycan-quaternary ammonium complexes, II. The effects of polymer molecular weight and sulfation.
Previous results have shown the possibility for obtaining high-resolution separations of glycosaminoglycans by partition in butanol/aqueous two-phase systems containing quaternary ammonium salts. In this paper, the effects on partition behavior of both polymer molecular weight and sulfation were examined. Two series of fractionated chondroitin sulfate polymers were isolated in which the molecular weight and sulfation varied systematically. In the molecular weight series the six samples, spanned the range from 3200 +/- 300 to 19 700 +/- 500 and each sample carried 0.8 sulfate groups per uronic acid residue. In the sulfation series, each sample had an essentially constant molecular weight of 13 000, but the sulfation varied from 0.58 to 0.88 sulfate groups per uronic acid. The C50 of each of these samples was determined in the 1-butanol/aqueous NaCl phase system containing 1% hexadecylpyridinium chloride. In the series wherein the molecular weight varied, the C50 increased with molecular weight up to 12 000 where a limiting value was reached. In the series wherein the sulfation varied, a linear relationship was found between the C50 and the square of the number of anionic substituents per disaccharide. These results show that fractionation by partition techniques will be sensitive to the anionic nature of the polymer, but for the common connective tissue glycosaminoglycan, there will be no fractionation according to molecular weight.
D002809:Chondroitin Sulfates / Q000302:isolation & purification; D006025:Glycosaminoglycans / Q000302:isolation & purification; D006801:Humans; D008970:Molecular Weight; D011485:Protein Binding; D000644:Quaternary Ammonium Compounds
D006025:Glycosaminoglycans; D000644:Quaternary Ammonium Compounds; D002809:Chondroitin Sulfates