Comparative bioactivity and phytochemical profiling of natural and processed honey: An integrated experimental and in silico approach

Abstract

Honey is widely recognized for its nutritional and therapeutic properties, which vary with origin and processing methods. This study compared the phytochemical composition, antioxidant activity, α-amylase inhibition, trace metal content, and proximate composition of natural and processed honey. HPLC analysis revealed higher levels of phenolics and flavonoids in natural honey. Antioxidant assays (DPPH and FRAP) confirmed significantly greater radical scavenging and reducing power in natural honey. Although processed honey showed stronger α-amylase inhibition at lower concentrations (85 % inhibition at 500 µg/mL), its activity declined at higher doses. Trace metal analysis indicated that natural honey was richer in essential elements such as potassium, manganese, and chromium, while processed honey contained higher levels of toxic metals like lead and iron. Molecular docking revealed that honey-derived compounds-Rutin, Saponin, and Naringenin-exhibited strong α-amylase binding affinities (-10.2 to -10.8 kcal/mol), comparable to Acarbose (-12.5 kcal/mol). Molecular dynamics simulations further confirmed that naringenin and saponin formed the most stable and rigid α-amylase complexes, as evidenced by low RMSD and RMSF values, indicating strong and persistent interactions. Overall, natural honey provides superior antioxidant, nutritional, and structural benefits, reinforcing its therapeutic potential in oxidative stress management and glycemic regulation.