This compound is primarily utilized as a chromogenic substrate in biochemical and enzymatic assays. It is commonly employed to detect and measure the activity of β-glucosidase enzymes. When the enzyme acts on this substrate, it cleaves the glucosidic bond, releasing 2-chloro-4-nitrophenol, which is a yellow-colored product. The intensity of the yellow color, measured spectrophotometrically, is directly proportional to the enzyme activity. This makes it a valuable tool in research and diagnostic laboratories for studying enzyme kinetics, screening enzyme inhibitors, and assessing enzyme-related disorders. Additionally, it is used in environmental studies to evaluate the presence and activity of β-glucosidase in soil and water samples, aiding in the assessment of microbial activity and organic matter decomposition.