The protein fractions were subjected to acetylation using the synthesized Aspirin-NHS. The reaction was conducted in a simulated cellular environment using 1X phosphate buffered saline to achieve a pH of 7.4. Aspirin-NHS was added to achieve a 5 times molar excess. The reaction mixture was incubated at a controlled temperature of 37°C for 1 hour, to facilitate optimal acetylation. Following the acetylation period, the reaction was quenched by adding hydroxylamine to a final concentration of 1 M, effectively arresting further acetylation reactions. Acetylated proteins were collected for further analysis through SDS-PAGE and mass spectrometry.