The main causative factors in the development of pulpal and periapical inflammation are bacteria, and the success of endodontic treatment depends on the eradication of microorganisms from the root canal system and prevention of reinfection Objective: To evaluate the antimicrobial efficacy of chemical versus herbal intracanal medicaments against Enterococcus faecalis biofilm. Method: Agar diffusion test was performed with Propolis (Group A), 2% Chlorhexidine (Group B), N-acetyl cysteine (Group C) and combination of 2%CHX+NAC (Group D) against planktonic cells. The diameter of zones bacterial inhibition were measured (mm) and recorded for each solution. For biofilm formation thirty freshly extracted teeth were vertically sectioned into two halves resulting in total of 60 samples.The samples were inoculated with bacterial suspension and incubated at 37°C for 2 weeks for biofilm formation. The samples were then divided into four experimental groups with 15 samples in each group. The samples were gently washed in saline and placed in culture wells containing the test solutions, i.e., Propolis, 2% CHX, NAC, a combination of 2% CHX and NAC in 1:1 ratio. Biofilm formed on root canal surface were removed using sterile scalpel and inoculated in 1ml of BHI broth, and mixed with the test medicament in equal proportions (1ml each). Spread plate method was performed in an interval of 2 hrs starting from 0- 24 hrs. to check the formation of E.faecalis colonies. The effect of medicament on the biofilm were expressed in terms of percent viability.