The enzyme hyaluronidase from human synovial fluid was partially purified using gel filtration and chromotography on a hydroxylapatite column. A 4.1 fold purification of hyaluronidase having a specific activity of 1.1 x 10 to the minus 4th power micro molminmg was obtained. The enzyme was inhibited by the gold complex Auranofin, and the steroid derivative, 22 ketocholesterol oxime. It was shown that the complexes differed in their ability to inhibit the enzyme. The rate constants and equilibrium inhibition constants were calculated. Kinetic analyses demonstrated an apparent uncompetitive inhibition by the steroid derivative 22-ketocholesterol oxime.