Introduction:Irrigants remove or dissolve smear layer formed during instrumentation. Thus it is important to study the effect of different irrigant solutions on smear layer removal.Objectives:The aim of this study is to determine which irrigant effectively removes the smear layer from the coronal, middle, and apical third of the root canal.Materials and Methodology:One hundred and twenty single-rooted mandibular premolars were decoronated and biomechanical preparation was done through hand instrumentation up to size 40 k file with 2.5% NaOCl irrigation between each successive filing, followed by irrigation with 5 ml of saline. The teeth were divided into Groups I, II, III, and IV containing 30 samples each and irrigated with 5 ml of 17% ethylenediamine tetraacetic acid (EDTA), 10% Citric acid, 7% Maleic acid and normal saline respectively for 1 min and final irrigation was done with 5 ml of distilled water of each sample. The canals were dried with 2% absorbent paper points. The roots were then split with a chisel and mallet. One-half of each tooth was selected and then was analyzed using a scanning electron microscope. The dentinal surfaces were observed at the cervical, middle, and apical third with ×2000 for the smear layer. The scores were attributed, according to Torabinejad et al. rating system.Results:7% maleic acid and 10% citric acid both are equally effective in smear layer removal from coronal and middle third, but in apical third 7% maleic acid is more effective than 10% citric acid. Between citric acid and EDTA, both are equally effective in smear layer removal from coronal and middle third, but in apical third, 10% citric acid is more efficacious than 17% EDTA.Conclusion:Within the limitations of the study, it can be concluded that all three tested irrigants removed the smear layer from coronal, middle, and apical third. However, in apical third 7% maleic acid is the most efficacious irrigant in smear layer removal.