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Hyaluronic Acid

Hyaluronic acid (Hy) is a highly polymerized mucopolysaccharide present in the highest concentrations in mammals in vitreous humor of the eye, human umbilical cords and joint fluids (Myer, K. and M.M. Rapport, Advances in Enzymology, 13, 199, 1952). It is one of the constituents of ground substance. It may be isolated in widely ranging degrees of purity and polymerization (Kaye, M.A.G. and M. Stacey, Biochem. J., 48, 249, 1951). Hyaluronic acid consists of a polymer of N-acetylglucosamine and glucuronic acid probably arranged as alternating units in a flexible chain. The Hy molecule is negatively charged but the degree of charge varies greatly with the ionic environment. This negative charge results in the mutual precipitation of cationic proteins such as albumin at low pH as in the Hyaluronidase assay procedure.

We define the purity in terms of a preparative definition. The highly polymerized nature of the material is established in terms of the limiting viscosity number [h]-the level of which exceeds for all grades.

Protein determinations are carried out by the method of Lowry (Lowry, A.L., et al, J. Biol. Chem., 193, 265, 1951). Chondroitin sulfate is detected on electrophoresis on cellulose acetate using Alcian Blue as dye. The homogeneity of Hyaluronic acid is further confirmed by chromatography on DEAE-cellulose (Schmidt, M., Biochim. Biophys. Acta, 63, 346, 1962; Berman, E.R., Biochim. Biophys. Acta, 58, 120, 1962).

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