To evaluate the influence of passive ultrasonic irrigation (PUI) to remove the smear layer with different irrigation solutions. Seventy-eight single rooted mandibular premolars were used. The working lengths were measured and the root canals were prepared by using nickel titanium rotary files. The specimens were subjected to six different irrigation regimens: sodium hypochloride (NaOCl) with classic irrigation (CI); NaOCl with PUI; EDTA with CI; EDTA with PUI; QMix with CI; and QMix with PUI. The specimens were split longitudinally. Scanning electron microscopy images were taken to evaluate the amount of residual smear layer and scored. Statistical analysis was performed with the Kruskal-Wallis test. The Dunn's test was used for post hoc testing after the Kruskal-Wallis test. The smear layer scores of apical thirds were significantly higher than coronal and middle thirds except for NaOCl + CI group (p < 0.001). The irrigation with PUI improved the efficacy of all irrigation solutions on removal of smear layer in both coronal and middle thirds (p < 0.001). The PUI activation increased the activity of NaOCl, EDTA, and QMix, respectively without any significance. All irrigation regimens were significantly more effective in coronal and middle thirds compared to apical third. PUI significantly increased the efficacy of smear layer removal regardless of irrigation solution.