A new chromatographic method for the fractionation of hyaluronic acid

Sodium hyaluronate from rooster comb, umbilical cord, bovine vitreous and a special commercial hyaluronic acid preparation, "HealonR" have been fractionated at 4 degrees C on DEAE-Sephacel columns using salt gradients from 0 leads to 0.4M NaCl. All samples examined displayed a high degree of polydispersity. The number of well separated fractions observed in these chromatograms ranges from 10-30 depending on the source of the material. A molecular weight distribution effect does not seem to be the main factor controlling this fractionation as no differences could be observed when the limiting viscosity numbers (eta) for numerous fractions isolated from the same sample were compared with each other. As no galactosamine, sulfate or phosphate is found directly linked to these fractions, it is suggested that disaggregation of an hyaluronate molecular aggregate could be the basis for this fractionation.