The main aim of this study is to utilize the chromogenic substrate BOC-Val-Leu-Gly-Agr-pNA, the commercially available LAL reagent was altered to develop a kinetic chromogenic LAL reagent. With the help of Zwittergent 3-14, an LAL reconstitution buffer sensitive to endotoxins was produced. A commercially available EIA QuantTM ELISA microplate reader was customized to carry out kinetic measurements to determine the time taken to attain the optical density of the samples. Plotting a standard line as the log of the time required to reach the optical density (seconds) against the log of the endotoxin concentration (EU/mL), it was found that the least two dilution out of 5 serial dilutions were not detectable in the modified microplate reader.