3,4-Dinitrophenyl N-acetyl-beta-D-glucosaminide, a synthetic substrate for direct spectrophotometric assay of N-acetyl-beta-D-glucosaminidase or N-acetyl-beta-D-hexosaminidase.

3,4-Dinitrophenyl N-acetyl-beta-D-glucosaminide (3,4-dnpGlcNAc) was synthesized and proposed as an artificial substrate for N-acetyl-beta-D-glucosaminidase (EC 3.2.1.30) and N-acetyl-beta-D-hexosaminidase (EC 3.2.1.52) (collectively abbreviated as NAGase). 3,4-dnpGlcNAc is water soluble and is fairly stable in aqueous medium. 3,4-Dinitrophenol released from 3,4-dnpGlcNAc by NAGase absorbs light of 400 nm at pH near 5, where the activity of urinary NAGase is assayed for diagnostic use. Direct and sensitive spectrophotometric assay of NAGase is, thus, possible using 3,4-dnpGlcNAc as an artificial substrate by monitoring the increase of A400 due to the release of 3,4-dinitrophenol in a thermostated spectrophotometer.