Isolation of oligosaccharides enzymatically produced from hyaluronic acid.

In earlier work application of carbon column chromatography to the products resulting from hydrolysis of umbilical cord hyaluronic acid by testicular hyaluronidase yielded fractions differing in average molecular weight (2). Monosaccharides were not detected in any of the digests, equivalence of hexosamine and uranic acid content was demonstrated in all fractions, and the constancy of certain end-group values was established. It was concluded that purified preparations of the enzyme hydrolyzed the polymer to oligosaccharides, in agreement with earlier workers, and that only the glucosaminidic linkages were affected, the uronidic linkages remaining intact. The present report is concerned with the application of ion exchange and paper chromatography to the isolation of the individual oligosaccharides, and with the determination of their molecular sizes, as well as with the development of analytical methods suitable for their estimation in mixtures. The oligosaccharides isolated are designated oligosaccharides I to VII, in order of increasing size.