Mechanistic Determinants of Antibacterial Potency in Libyan and Saudi Monofloral Honeys: Integrated H₂O₂, Acidity, and Phenolic Chemistry

Abstract Background: Honey exhibits broad-spectrum antimicrobial activity through hydrogen peroxide, phenolics, and acidity. In the era of antimicrobial resistance (AMR), such multifactorial activity offers a natural strategy less prone to resistance development. Comparative insights into Libyan and Saudi honeys are limited, especially regarding mechanistic pathways. Methods: We analyzed 180 monofloral honey samples (Libyan: Sidr, Athel, Hannon; Saudi: Sidr, Talh, Sumra). Physicochemical quality was assessed per Codex/IHC standards, bioactive chemistry via phenolic and antioxidant assays, and hydrogen peroxide kinetics. Antibacterial potency was evaluated against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa, including mechanistic perturbations with catalase, PVPP, and pH neutralization. Results: All honeys complied with Codex quality limits. Libyan Sidr contained the highest phenolics and antioxidant activity, while Saudi Talh and Sumra generated the most hydrogen peroxide. Antibacterial profiles reflected these mechanistic differences: peroxide-dominant Talh/Sumra achieved the lowest MICs against S. aureus (10–12.5% w/v), Sidr honeys showed phenolic-driven intermediate potency, and Athel/Hannon relied on acidity. Conclusion: This is the first integrated mechanistic comparison of Libyan and Saudi honeys, directly linking floral origin to antibacterial pathways. The findings highlight region-specific translational potential: Saudi Acacia honeys as peroxide-driven candidates for topical wound care, and Libyan Sidr honeys as phenolic-rich nutraceuticals. By clarifying these mechanisms, the study contributes to natural antimicrobial discovery and supports global efforts against AMR.