Objectives: This study comparatively evaluated the efficacy of self-adjusting file (SAF), Endovac, and CanalBrush irrigant agitation protocols in removing calcium hydroxide (Ca(OH)<inf>2</inf> ) from the root canals. Materials and Methods: Sixty extracted human mandibular canine teeth were instrumented with ProTaper rotary instruments to size #40 and dressed with Ca(OH)<inf>2</inf>. The roots were randomly assigned to four groups according to irrigant agitation protocol used (n = 15). In Group 1: Conventional syringe irrigation (no activation, control); Group 2: Rotary brush agitation (CanalBrush); Group 3: Apical negative pressure irrigation (EndoVac system); and Group 4: Sonic agitation (SAF) were used. Scanning electron microscopic (SEM) evaluation was done for assessment of Ca(OH)<inf>2</inf> removal in the coronal and apical thirds. Statistical analysis was performed by Wilcoxon and Kruskal-Wallis tests. Results: There were statistically significant differences among the groups (P = 0.218). A statistically significant difference was seen between the test groups in Ca(OH)<inf>2</inf> removal from the apical third of the canal (P < 0.05). In the coronal third, there was no difference between the groups (P > 0.05). The most efficient Ca(OH)<inf>2</inf> removal in apical third was recorded in Group 3 (EndoVac) and Group 4 (SAF) (P < 0.05). In Group 4 (sonic agitation), there was no significantly difference between Ca(OH)<inf>2</inf> removal in coronal and apical thirds. Conclusions: SAF and EndoVac showed significantly better performance than CanalBrush and conventional syringe irrigation in removing Ca(OH)<inf>2</inf> from apical third of the root canals.