Abstract
Because they are highly unsaturated, plant lipids are sensitive to oxidation and constitute a primary target of reactive oxygen species. Therefore, quantification of lipid peroxidation provides a pertinent approach to evaluating oxidative stress in plants. Here, we describe a simple method to measure upstream products of the peroxidation of the major polyunsaturated fatty acids in plants, namely, linolenic acid (C18:3) and linoleic acid (C18:2). The method uses conventional HPLC with UV detection to measure hydroxy C18:3 and C18:2 after reduction of their respective hydroperoxides. The described experimental approach requires low amounts of plant material (a few hundred milligrams), monitors oxidation of both membrane and free fatty acids, and can discriminate between enzymatic and non-enzymatic lipid peroxidation.
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Acknowledgments
We would like to acknowledge the input from our colleague, the late Christian Triantaphylides, who was very helpful in the optimization of the experimental method described in this article.
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Ksas, B., Havaux, M. (2022). Determination of ROS-Induced Lipid Peroxidation by HPLC-Based Quantification of Hydroxy Polyunsaturated Fatty Acids. In: Mhamdi, A. (eds) Reactive Oxygen Species in Plants. Methods in Molecular Biology, vol 2526. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2469-2_13
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DOI: https://doi.org/10.1007/978-1-0716-2469-2_13
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