Preparation, Component Identification, and Antioxidant Activity of Anthocyanins From <i>Punica granatum</i> “Tunisia”

ABSTRACT Anthocyanins, a major class of flavonoids with potent antioxidant activity, are abundant in pomegranate ( Punica granatum L.). In this study, anthocyanins were extracted from Punica granatum “Tunisia” and purified to over 70% purity. The major anthocyanin components were characterized by LC–MS/MS analysis. Subsequently network pharmacology and molecular docking suggested that anthocyanins may exert antioxidant effects mainly through the Keap1–Nrf2–HO‐1 signaling pathway. The antioxidant activity of the extract was confirmed by DPPH, ABTS, and FRAP assays. In LPS‐induced RAW264.7 cells, the anthocyanin extract significantly reduced ROS and NO production, enhanced SOD activity, promoted Nrf2 nuclear translocation, downregulated Keap1, and upregulated HO‐1 expression. These results demonstrated that pomegranate anthocyanins exhibited strong antioxidant activity and may serve as promising natural antioxidants for functional food applications.