ObjectivesThis study evaluated smear layer removal by different chemical solutions used with or without ultrasonic activation after post preparation.Materials and MethodsForty-five extracted uniradicular human mandibular premolars with single canals were treated endodontically. The cervical and middle thirds of the fillings were then removed, and the specimens were divided into 9 groups: G1, saline solution (NaCl); G2, 2.5% sodium hypochlorite (NaOCl); G3, 2% chlorhexidine (CHX); G4, 11.5% polyacrylic acid (PAA); G5, 17% ethylenediaminetetraacetic acid (EDTA). For the groups 6, 7, 8, and 9, the same solutions used in the groups 2, 3, 4, and 5 were used, respectively, but activated with ultrasonic activation. Afterwards, the roots were analyzed by a score considering the images obtained from a scanning electron microscope.ResultsEDTA achieved the best performance compared with the other solutions evaluated regardless of the irrigation method (p < 0.05).ConclusionsUltrasonic activation did not significantly influence smear layer removal.