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A new type of chemiluminogenic substrate for N-acetyl-beta-D-glucosaminidase, o-aminophthalylhydrazido-N-acetyl-beta-D-glucosaminide, was prepared by incorporating an enzyme-removable substituent, N-acetyl-D-glucosaminide group, into the hydrazide moiety of luminol. This substrate releases luminol upon enzymatic hydrolysis. The enzyme was detected chemiluminescently using this substrate.

A chemiluminogenic substrate for N-acetyl-.BETA.-D-glucosaminidase, o-aminophthalylhydrazido-N-acetyl-.BETA.-D-glucosaminide.