This study aimed to compare the effects of different irrigants on root dentine microhardness, erosion and smear layer removal. A total of 72 root dentine slices were divided into six groups, according to the final irrigants used: Group 1: 17% ethylenediamine tetra-acetic acid (EDTA) + 2.5% NaOCl, Group 2: 7% maleic acid (MA) + 2.5% sodium hypochloride (NaOCl), Group 3: 1.3% NaOCl + mixture of tetracycline, acid and detergent (MTAD), Group 4: Smear Clear + 2.5% NaOCl, Group 5: 5% NaOCl, Group 6: saline. Vickers microhardness values were measured before and after treatment. In total, 42 root-halves were prepared for scanning electron microscope to evaluate the amount of smear and erosion in the coronal, middle and apical thirds. Data were analysed using two-way ANOVA, Duncan and two-proportion z-tests. Maleic acid showed the greatest reduction in dentine microhardness (P < 0.05), followed by EDTA and MTAD. EDTA, maleic acid, MTAD and Smear Clear removed smear layer efficiently in the coronal and middle thirds of root canal. However, in the apical region, maleic acid showed more efficient removal of the smear layer than the other irrigants (P < 0.05).