This compound is primarily used in biochemical research as a substrate for enzyme assays, particularly for studying the activity of esterase and lipase enzymes. When these enzymes act on it, they cleave the ester bond, releasing 4-nitrophenol, which can be easily detected and quantified spectrophotometrically due to its yellow color in alkaline conditions. This makes it a valuable tool for kinetic studies and enzyme characterization in laboratory settings. Additionally, it is employed in the development of biosensors and diagnostic kits for detecting enzyme activity in biological samples. Its stability and ease of detection make it a practical choice for high-throughput screening in pharmaceutical and biochemical applications.