Walnut oil was extracted using three different eco-friendly extraction methods, solvent extraction (using ethyl acetate [EA] and ethanol [ET]), aqueous enzymatic extraction (AEE), and ultrasound-assisted enzymatic extraction (UAEE), and their lipid yield, lipid composition, physicochemical analysis, mineral composition, total phenols, antioxidant capacity, and antimicrobial activity were analyzed and compared. The AEE technique offered a greater yield (50.6%) than the other extraction methods and gave comparatively higher linoleic acid (66.12%) content. Palmitic, oleic, linoleic, linolenic, and stearic acids were the principal components that GC/MS detected in all the oil samples. UAEE produced the most polyphenols (0.49 mgGAE/g), followed by AEE (0.46 mgGAE/g), EA (0.45 mgGAE/g), and ET (0.35 mgGAE/g). The DPPH assay results were in the order of UAEE (191 μmolTE/kg) > AEE (186 μmolTE/kg) > EA (153 μmolTE/kg) > ET (130 μmolTE/kg). The FRAP assay findings showed a similar pattern: UAEE (112 molTE/kg) > AEE (102 molTE/kg) > EA (96 molTE/kg) > ET (82 molTE/kg). Results suggested that for a higher extraction yield, AEE is the better technique and UAEE is the recommended method for enhancing walnut oil antioxidant capacity. Additionally, it was found that polyphenols considerably increased the antioxidant capacity of walnut oil and are thought to be health-promoting. The results demonstrated the antibacterial effectiveness of the extracted oil against Bacillus subtilis, Bacillus licheniformis, and Staphylococcus aureus. This study provides information about low-cost and ecofriendly technologies of walnut oil extraction for food, cosmetic, and medical uses.
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