FLUOROMETRIC DETERMINATION OF HYALURONIDASE AND CU(11), FE(11), AND CYANIDE ION INHIBITORS.
Technical rept. Apr-Jun 66,
EDGEWOOD ARSENAL MD
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A fluorometric method is described for the assay of the enzyme hyaluronidase and of cupric, ferrous, and cyanide ions, which inhibit the enzymic activity. The method for hyaluronidase is based upon the hydrolysis of nonfluorescent indoxyl acetate by the enzyme to give the highly fluorescent indigo white. The metal ions can be determined by recording their inhibitory effect on the enzymic activity. By this procedure, from 0.0010 to 0.0330 microgramsml of hyaluronidase, 0.10 to 4.0 microgramsml of CN-, and 0.20 to 12 microgramsml of Fe or Cu could be determined with standard deviations of plus or minus 1.8 for hyaluronidase and plus or minus 2.3 for the inhibitors. Evidence is offered to show that the activity of hyaluronidase, and not esterase, is measured. There appears to be a good correlation between the fluorometric method and other methods for hyaluronidase, and this technique should be a good one for the rapid, sensitive determination of this enzyme and of cupric, ferrous, and cyanide ions, which selectively inhibit its activity. Author