Further observations on the preparation and composition of the hyaluronic acid complex of ox synovial fluid

Abstract

Methods of separating hyaluronic acid from ox synovial fluid were re-investigated, using the variation of viscosity with velocity gradient as a criterion of degradation. Precipitation as mucin and precipitation by ethanol causes degradation, while ultrafiltration does not. The course of purification by ultrafiltration was followed. It was confirmed that the protein content of the complex is reduced to a limiting value of 25%. Electrophoresis was shown not to give products of lower protein content. Analysis of the product of ultrafiltration shows that 93% of its weight is accounted for by acetyl hexos-amine, hexuronic acid, protein and ash. The protein content of the complex was found to be lower than was previously reported. A revised value was given for the composition of mucin prepared on a small scale. Degradation by hyaluronidase was shown to leave a considerable fraction of the material in a polymerized state (molecular wt. of the order of 105). The ultracentrifugal sedimentation of purified hyaluronic acid complex was re-investigated. Slight modifications of the values of physical quantities were given, based on the revised analytical values and sedimentation constant. Conclusions about the composition and molecular configuration of the hyaluronic acid complex were summarized.