This compound is primarily utilized in biochemical research as a substrate for the assay of trypsin-like enzymes. It helps in studying enzyme kinetics and activity, particularly in the context of proteolytic enzymes. Researchers employ it to monitor the hydrolysis process, where the enzyme cleaves the substrate, releasing 4-nitroaniline, a chromogenic product measurable by absorbance at around 405 nm. This application is crucial in understanding enzyme mechanisms and in the development of enzyme inhibitors, which can have therapeutic potential. Additionally, it is used in diagnostic laboratories to assess enzyme levels in biological samples, aiding in the diagnosis of certain medical conditions. Its role in these studies contributes significantly to advancements in biochemistry and medical science.