Aim: The aim of the present study was to compare the antimicrobial efficacy of various intracanal medicaments against Enterococcus faecalis (E. faecalis) and Candida albicans (C. albicans). Materials and methods: Sixty single rooted, non-carious, permanent mandibular premolars with no developmental defects that were previously extracted for the orthodontic purpose were included in this study. E. faecalis and C. albicans strains were cultured on brain heart infusion (BHI) and sabouraud dextrose (SD) agar plates respectively. Sixty specimens were divided into two main groups and three subgroups with 10 teeth receiving a medicament; group I: E. faecalis (Subgroup Ia: BioPure MTAD, Subgroup Ib: propolis, Subgroup Ic: Triple antibiotic paste), group II: C. albicans (Subgroup IIa: BioPure MTAD, Subgroup IIb: propolis, Subgroup IIc: Triple antibiotic paste). The antimicrobial activity of the intracanal medicaments against E. faecalis and C. albicans were assessed at the end of 2nd and 7th day. The mean zones of inhibition were analyzed with a one-way ANOVA test. Results: After 2 days, the mean zone of inhibition of E. faecalis and C. albicans was maximum for triple antibiotic paste (24.74 ± 0.622 mm, 28.22 ± 0.489 mm), followed by BioPure MTAD (19.58 ± 1.734 mm, 24.75 ± 0.954 mm) and propolis (13.10 ± 0.278 mm, 17.96 ± 0.163 mm). Similarly, the mean zone of inhibition of E. faecalis and C. albicans was maximum for triple antibiotic paste (26.86 ± 0.112 mm, 32.10 ± 0.908 mm), followed by BioPure MTAD (20.13 ± 1.842 mm, 27.22 ± 1.977) and propolis (14.11 ± 0.101 mm, 19.90 ± 0.742 mm) after 7 days. Statistically significant differences (p <0.0001) were found between the groups. Conclusion: Present study concluded that the antimicrobial effectiveness of triple antibiotic paste was significantly more than BioPure MTAD and propolis against E. faecalis and C. albicans at the end of 2nd and 7th day. Clinical significance: An endodontic treatment is considered successful only when the root canals are completely eradicated of microorganisms. As the morphology of the root canals is complex, mechanical instrumentation alone cannot completely debride them. Thus, the role of intracanal medicaments in complementing the mechanical instrumentation becomes important.