Analysis of heparin origin by HPLC quantitation of disaccharide components

A simple method was developed to analyze by high performance liquid chromatography unsaturated disaccharide isomers, derived from heparin by enzymatic digestion. This method was successfully exploited in the investigation of heparin origin. The percent amount of 6-sulphated disaccharides in the heparin extracted from porcine mucosa was found to be higher than that contained in the heparin extracted from bovine mucosa. The differences in the contents of the disaccharides obtained by enzymatic beta-elimination cleavage of heparin were confirmed by 13C-NMR measures of heparin in toto. The processes for extracting and purifying heparin may, however, modify the sulphation pattern of heparin. The structure of the latter seems to depend on the species owing to the specificity of the biosynthesis.