Abstract
Hyaluronidase activity has been measured by a semimicro radiochemical method. 3H-Hyaluronic acid was prepared using hydrazine to remove some of the acetyl groups followed by reacetylation with tritiated acetic anhydride. Hydrolysis was carried out using an extract from rat liver lysosomes and a cruder extract from other tissues. At the completion of hydrolysis, cetylpyridinium chloride was added to precipitate unreacted substrate. Smaller molecular-weight oligosaccharides produced by hydrolysis remain in solution to be determined by liquid scintillation counting. This assay gives results that are similar to those obtained by the assay using N-acetylglucosamine end-group determination but, in the case of tissue extracts, in 1 hr instead of 18 hr.
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