BACKGROUND 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. OBJECTIVE 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. METHODS 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. CONCLUSIONS 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.