The interactions between protein and ligand are crucial to practically every biological function in the body, and investigations of these interactions are critical from the perspective of logical medication development. In this work, several biophysical methods like UV–Visible spectroscopy, fluorescence spectroscopy, and in-silico studies have been explored to examine the interaction between aceclofenac and haemoglobin. Spectrofluorimetric data confirmed the static nature of the protein quenching mechanism caused by the medication. ΔG for the formation of the complex via the interaction was found to be − 12.65 kJ/mol. Molecular docking showed a strong interaction between aceclofenac and hemoglobin with a binding energy of − 31.79 kJ/mol. Further, molecular dynamics simulations using Gromacs were run to investigate the stability of the protein with and without aceclofenac, and the ΔG for the formation of the complex (hemoglobin-aceclofenac) is − 105.09 kJ/mol with an error of 13.51 kJ/mol. Spectroscopic and in-silico calculations indicate the feasible binding of the aceclofenac with hemoglobin. Further, the interaction of haemoglobin with aceclofenac was studied by cyclic voltammetry.