Phase-specific checkpoint modulation by organic and inorganic constituents of Berberis thunbergii DC. leaf extract in A549 lung cancer cells.

Medicinal plant extracts are typically interpreted mainly through their organic phytochemical constituents; however, growing evidence suggests that inorganic components may also contribute to biological activity and cellular fate. In this study, we performed an integrated organic–inorganic mechanistic evaluation of the anticancer activity of Berberis thunbergii DC. leaf extract, focusing on its effects in A549 human lung carcinoma cells. Treatment concentrations were determined using MTT-based cytotoxicity assays. The phenolic composition of the methanolic extract was characterized by LC–MS/MS, while apoptosis induction and cell-cycle distribution were analyzed by flow cytometry. Inorganic anions were quantified by ion chromatography under conditions corresponding to the A549 IC50 value. LC–MS/MS profiling identified rutin, chlorogenic acid, and hesperidin as the major phenolic constituents. Exposure of A549 cells to the extract at its IC50 significantly reduced cell viability, whereas HEL299 cells retained high viability. Ion chromatography revealed the presence of fluoride, phosphate, and sulfate anions. Functional assays using certified NaF, Na2HPO4, and Na2SO4 standards showed that NaF significantly induced apoptosis and G0/G1 arrest, while sulfate caused S-phase depletion and G2/M accumulation. No significant effects were observed after phosphate treatment. These phase-specific responses suggest that Berberis thunbergii DC. leaf extract acts as a phytochemical–ionic system in which inorganic anions contribute to selective cell-cycle modulation in A549 cells.