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Objectives:To evaluate the efficacy of 35% orthophosphoric acid, 24% ethylenediaminetetraacetic acid (EDTA) and 10% citric acid in the removal of smear layer from retrocavities prepared with ultrasonic retro-tips using scanning electron microscopy.Materials and Methods:Root canals of fifty single-rooted teeth were cleaned, shaped, and obturated with gutta-percha and AH plus sealer. The apical 3 mm of each root was sectioned with a diamond disc and retrograde cavities were prepared with Kis # 1 ultrasonic retro-tips to a depth of 3 mm. Retro cavities in Group I were treated with a gel of 35% orthophosphoric acid for 15 seconds, Group II were treated with a gel of 24% EDTA at neutral pH for 2 minutes, and Group III were treated with a gel of 10% citric acid for 2 minutes, followed by 1 minute rinsing with distilled water for all groups. The samples were prepared for scanning electron microscopic observation. Scoring was performed for the presence of the smear layer on the walls of the retrocavity.Results:In the orthophosphoric acid group, it was observed that all dentinal tubules were open in 70% of the samples. The majority of analyzed samples in EDTA and citric acid group showed dentinal tubules covered with the smear layer.Conclusions:Application of 35% orthophosphoric acid gel for 15 seconds on retrocavities prepared with ultrasonic retro-tips is the most effective means for removal of smear layer.