This study was done to clinically investigate the efficacy of glycyrrhizin-based intracanal medication (ICM) compared to calcium hydroxide Ca(OH)2 and Ledermix on the bacterial counts inside the root canals of necrotic teeth with chronic periapical periodontitis. Thirty-six patients having single-rooted and single-canaled necrotic teeth with chronic periapical periodontitis were enrolled in this research. Aseptic control measures were taken before clinical steps. Access cavity preparation was done. First bacteriological samples (S1) were collected immediately after access cavity preparation and before cleaning and shaping inside the root canals. Cleaning and shaping were performed on the root canals. Patients were randomly allocated into 3 groups according to the type of intracanal medicament used [Ca(OH)2, Ledermix, Glycyrrhizin]. Second bacteriological samples (S2) were collected after 1 week from placing the ICMs. S1 and S2 were transferred to the lab of microbiology for culturing on blood agar dishes in anaerobic conditions, and the bacteria on the plates were enumerated as colony-forming units (CFUs) by the manual counting method. The anti-bacterial efficacy of the ICM was estimated by the percentage reduction in the bacterial colonies from S1 to S2. All tested ICM documented a significant reduction in the CFUs from S1 to S2 (p < 0.05). Regarding S1, there was no significant difference between the three tested materials (p > 0.05). Regarding S2, Glycyrrhizin and Ledermix showed significantly lower CFUs than Ca(OH)2. There was no significant difference between Glycyrrhizin and Ledermix. Under the conditions of the present study, Glycyrrhizin and Ledermix showed comparable antimicrobial effects that were better than the commonly used Ca(OH)2 ICM. Glycyrrhizin may be a promising ICM. Herbal medicine can be considered as an alternative antimicrobial material for root canal disinfection because it has many benefits, like low toxicity, absence of microbial resistance, and favorable antimicrobial efficacy. This study highlights the clinical efficacy of Glycyrrhizin as a promising ICM. How to cite this article: Eltantawi AR, Abdel-Razik GM, Elhawary YM, et al. Efficacy of Glycyrrhizin as an Intracanal Medicament on Bacterial Load Reduction in Primary Infected Root Canals: A Randomized Clinical Trial. J Contemp Dent Pract 2024;25(6):540-546.