Background. Afzelia africana is a plant species with well-documented ethnobotanical and medicinal properties. The plant is reported to have various secondary metabolites and had been applied for the treatment of various diseased conditions. Objectives. The study objectives include fractionation, isolation, purification, and characterization of eriodictyol from the bark of A. africana, and the determination of its antimicrobial and antioxidant activities. Methodology. The series of methodologies that were employed include fractionations and purification (column chromatography), characterization (HPLC, LC-MS, IR, 1H, 13C, DEPT-135, HSQC, and HMBC), antimicrobial assays (microbroth dilution and checkerboard assay), and antioxidant activities assays (ABTS and DPPH scavenging capacity). Results. The study reports the identification and characterization of eriodictyol from the bark of A. africana which exhibited potent antioxidant activities against ABTS and DPPH radicals with scavenging capacities (SC50) of 2.14 ± 0.05 and 2.51 ± 0.06 µg/mL, respectively. The compound exhibited its antimicrobial activity by reporting good bacteriostatic activities (MBC/MIC > 4) against Staphylococcus aureus (SA), methicillin-resistant Staphylococcus aureus (MRSA), and fluconazole-resistant Candida albicans (CA2). Moreover, a broad spectrum of bactericidal effects (MBC/MIC ≤ 4) was reported against Streptococcus mutans (SM), Escherichia coli (EC), Bacillus subtilis (BS), Klebsiella pneumonia (KP), Pseudomonas aeruginosa (PA), Salmonella typhi (ST), and standard Candida albicans (CA1). The compound further exhibited synergistic effects against EC, KP, ST, and MRSA; ST; and CA2 when combined with ciprofloxacin, tetracycline, and nystatin, respectively. However, antagonistic effects were observed against PA and CA1 when combined with ciprofloxacin and ketoconazole, respectively. Conclusion. The study reports for the first time the identification of eriodictyol from the bark of A. africana which exhibited significant antioxidant and antimicrobial properties.