Beeswax Waste-Derived Pyrolysis Liquid as a Novel Multifunctional Bioactive Fraction: An Integrated Chemical and Biological Evaluation

In this study, beeswax waste, a low-value by-product of apiculture, was converted into a liquid fraction with bioactive potential through pyrolysis-assisted thermochemical transformation. The LC–MS/MS-based chemical profile, total phenolic content (TPC), total flavonoid content (TFC), antioxidant responses, and antibacterial, antidiabetic, anti-inflammatory, and anticholinesterase activities of beeswax waste pyrolysis liquid (BWPL) were investigated. The TPC and TFC values were determined as 95.30 ± 1.13 mg GAE/L and 51.67 ± 1.12 mg QE/L, respectively. LC–MS/MS analysis revealed benzoic acid and vanillin as the predominant compounds, along with various phenolic acids and flavonoid derivatives. BWPL showed 51.83 ± 1.51% inhibition in the ABTS•⁺ assay and 9.71 ± 0.28% inhibition in the DPPH assay. It also exhibited measurable redox capacity in Fe3+ and Cu2+ reducing power assays. In antibacterial assays, BWPL produced inhibition zones against all tested strains at 1 mg/mL and against E. coli and E. faecalis at 500 µg/mL. The IC50 values of BWPL for 5-LOX and α-glucosidase were 38.17 µg/mL and 1503.0 µg/mL, respectively. In addition, the IC50 values for AChE and BChE were calculated as 112.38 ± 0.87 mg/mL and 13.01 ± 0.42 mg/mL, respectively. PCA and correlation analyses indicated the multidimensional relationships between the LC–MS/MS-based chemical profile and biological activities. Overall, the pyrolytic conversion of beeswax waste into a chemically complex liquid fraction provides a preliminary scientific basis for waste valorization and for further investigation of pyrolysis-derived bioactive fractions.