Aims: The purpose of this study was to compare the effectiveness of various medicaments, including calcium hydroxide alone, calcium hydroxide with iodoform, calcium hydroxide points, calcium hydroxide with 2% chlorhexidine gel against Enterococcus faecalis in vitro. Methodology: Seventy five extracted single-rooted human maxillary teeth were used. After removing the crown, each root was instrumented up to size 50 by using a conventional technique and were infected with E faecalis. Subsequently, the roots were divided into 4 treatment groups: group 1 was treated with calcium powder hydroxide, group 2 was treated with calcium hydroxide with iodoform, group 3 was treated with calcium hydroxide points, and group 4 was treated with calcium hydroxide with 2% chlorhexidine gel, group 5 was treated with 0.9% sterile saline serving as negative control. Microbial samples were taken after 7, 15, and 30 days. After incubation, dentine chips were obtained from each root canal and examined microbiologically. The microbiological samples were plated to count colony-forming units in per milligram of dentin. Results: The results showed that the calcium hydroxide with 2% chlorhexidine gel was significantly more effective than calcium hydroxide alone, calcium hydroxide with iodoform, calcium hydroxide points and control saline solution. Conclusion: Under the conditions of this study, calcium hydroxide with 2% chlorhexidine gel is effective in the elimination of E faecalis from the root canal system. However, to support this in vitro observation, further in vivo studies are needed. Key words: Calcium hydroxide, Chlorhexidine