Aim: The present study compares and assesses the effectiveness of 0.2% Chitosan nanoparticle solution, 18% Etidronic acid and 17% EDTA in removing smear layer using SEM image analysis. Materials and Methods: Twenty freshly extracted mandibular premolars were used. Following biomechanical preparation, samples were divided into Group A(0.2% chitosan nanoparticle solution), Group B(18% Etidronic acid), and Group C(17% EDTA), and Group D(0.9% Saline) containing 5 samples each. After irrigation the samples were sectioned longitudinally and were subjected for SEM analysis at apical, middle, and coronal levels. Hullsman's criteria were used to assign scores to the images. The statistical analysis was conducted using Kruskal Wallis test, Mann Whitney – U test, with a significance level of p< 0.05. Results: When 0.2% chitosan solution and 18% Etidronic acid were used as the final irrigation, smear layer removal in the coronal, middle, and apical thirds was more successful. All of the irrigants had weak smear layer removal properties at the apical third, whereas 0.2% chitosan was significantly better followed by 18% Etidronic acid. Conclusions: Compared to 17% EDTA and saline irrigants, 0.2% chitosan nanoparticle solution was more successful in eliminating the smear layer followed by 18% Etidronic acid