Decoding Ferroptosis: Mechanistic Insights and Future Directions in the Precision Analysis of Lipid Hydroperoxides

Abstract Ferroptosis is a regulated form of cell death driven by iron-dependent lipid peroxidation of polyunsaturated phospholipids. While its core regulatory axis involving glutathione peroxidase 4 (GPX4) and glutathione has been extensively studied, emerging evidence highlights the presence of multiple parallel defense systems. In addition, recent findings identify peroxiredoxin 6 as a critical regulator of intracellular selenium utilization, linking selenium metabolism to GPX4 biosynthesis and ferroptosis sensitivity. Despite these advances, a comprehensive understanding of ferroptosis mechanisms requires precise characterization of lipid hydroperoxides, the primary executioners of this cell death pathway. Here, we review the molecular basis of ferroptosis and its multilayered defense networks, with a particular focus on analytical challenges in lipid peroxidation research. We highlight recent progress in mass spectrometry–based approaches, including alkali metal adduction strategies, which enable positional isomer–resolved analysis of lipid hydroperoxides.