The aim of this study was to investigate in vitro the antimicrobial activity of 0.2%, 1%, and 2% chlorhexidine gluconate (CHX gel and CHX liquid), against endodontic pathogens and compare the results with the ones achieved by 0.5%, 1%, 2.5%, 4%, and 5.25% sodium hypochlorite (NaOCl). A broth dilution test was performed, and the timing for irrigants to kill microbial cells was recorded and statistically analyzed. Both 2.0% gel and liquid formulations eliminated Staphylococcus aureus and Candida albicans in 15 seconds, whereas the gel formulation killed Enterococcus faecalis in 1 minute. All tested irrigants eliminated Porphyromonas endodontalis, Porphyromonas gingivalis, and Prevotella intermedia in 15 seconds. The timing required for 1.0% and 2.0% CHX liquid to eliminate all microorganisms was the same required for 5.25% NaOCl. The antimicrobial action is related to type, concentration, and presentation form of the irrigants as well as the microbial susceptibility.9717984