The aim of the current study was to assess the cleaning and smear layer removal efficacy of two different rotary files with or without chemical agents on primary teeth. For the study, 90 extracted primary maxillary incisors without internal or external resorption and with at least two-thirds of complete roots were chosen. Then, based on the kind of instruments used to clean and shape the canals, they were randomly assigned to three experimental groups, each consisting of 30 teeth. Group-I: The canal was instrumented manually with K-files, Group-II: The canal was instrumented with Kedo-S files, Group-III: The canal was instrumented with Kedo-SG Blue files. After the canals were finally instrumented, 2 mL of QMixTM solution was used to irrigate 15 samples from each group. The samples were subsequently allowed to remain in the canals for 90 seconds in order to eliminate the smear layer. After that a stereomicroscope was used to assess the cleaning effectiveness. With irrigant solution, the highest mean value was found in manual K-files (2.86 ± 0.34), followed by Kedo-S files group (1.34 ± 0.26) and Kedo-SG Blue files (1.28 ± 0.18). Without irrigant solution, the highest mean value was found in manual K-files (2.92 ± 0.22) followed by Kedo-S files group (1.44 ± 0.18) and Kedo-SG Blue files (1.36 ± 0.14). There was a statistically significant difference found at all the three levels. On conclusion, the current study's findings demonstrated that irrigation solution was significantly more effective in cleaning and removing smear layers from pediatric rotary files than manual K-files. The effectiveness of endodontic therapy depends on a successful chemomechanical preparation. The canals are instrumented using either hand files or rotary instruments; there are several irrigation and instrumentation techniques. In order to completely sterilize the canals, chemical agents are utilized for irrigation during instrumentation. Due to their numerous biological, antibacterial, anti-inflammatory, and antioxidant qualities, many natural compounds are also utilized as irrigants. How to cite this article: Abushanan A. Evaluation of the Smear Layer Removal Ability of Various Rotary Files with/without Chemical Agents on Primary Teeth: An In Vitro Study. J Contemp Dent Pract 2024;25(4):354-357.