Paper chromatography of heparin and related sulfated mucopolysaccharides

A paper chromatographic system was developed for the resolution of mixtures of sulfated mucopolysaccharides. The effect of varying the ammonium formate buffer/isopropanol ratio in the developing solution on the R F values of heparin and chondroitin sulfate is presented. A complete separation of heparin from chondroitin sulfate was obtained with three solvent mixtures containing different proportions of buffer and isopropanol. One of these solvent mixtures, ammonium formate buffer-isopropanol (65:35) v/v, resolved commercial beef heparin into two principal components, on “fresh” paper. This system was used to compare beef heparin with mouse mast cell tumor heparin, rat heparin, sheep lung heparin, chondroitin sulfate, and β-heparin. A common metachromatic component with an R F of approximately 0.5 was0found in all of the heparin preparations chromatographed with this solvent mixture. “Aging” of the filter paper resulted in the loss of the ability of the buffer-isopropanol mixture (65:35) v/v to resolve the beef heparin. Increase of the buffer-isopropanol ratio to (70:30) v/v restored the resolution of the beef heparin on “aged” paper. Heparitin sulfate and α-heparin monosulfate were compared to beef heparin using the latter system.