Tooth discoloration is a common concern associated with the use of intra-canal medicaments, particularly those containing antibiotics. This study aims to evaluate the tooth discoloration potential of an experimental antibiotic paste modified with Nano Chitosan (APNC) and compare it with two antibiotic pastes and two calcium hydroxide-based pastes over different time intervals. : Fifty bovine incisors were standardized and prepared up to size 60. The teeth were randomly assigned into five groups based on the medicament applied: Metapaste, Metapex, modified Triple Antibiotic Paste (mTAP), Double Antibiotic Paste (DAP), and APNC. A digital spectrophotometer was used to measure the color parameters (L*, a*, and b*) at two zones, above and below the cemento-enamel junction (CEJ), across four-time points: before application (T0- baseline), immediately after application (T1), after two weeks (T2), and after one month (T3). The color changes (ΔE) were calculated and statistically analyzed using repeated-measure ANOVA. : Statistically significant differences in discoloration were observed between the tested medicaments after one month ( < 0.05). mTAP caused the highest ΔE values both above and below the CEJ at all time points, particularly after one month ( < 0.05). Conversely, APNC, Metapaste, and DAP demonstrated the least discoloration, with no significant differences among them. The degree of discoloration was time-dependent and more pronounced below the CEJ in all groups. : After one month, the experimental APNC paste induced tooth discoloration comparable to that of Metapaste, indicating minimal esthetic compromise. APNC may be a promising alternative to traditional antibiotic pastes with minimal discoloration effects.