High Performance Liquid Chromatography Analysis of Reduced and Oxidized Glutathione in Woody Plant Tissues

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A rapid, and sensitive high performance liquid chromatography method for quantifying both the reduced and oxidized forms of glutathione simultaneously in woody plant tissues is reported. Samples were extracted in 10% perchloric acid with 1 mM bathophenanthroline disulfonic acid, homogenized, and centrifuged. The supernatant was carboxy-methylated with iodoacetic acid, and derivatized with 2,4-dinitro-1-fluorobenzene. A 3-aminopropyl-Spherisorb column was used for separations. Dinitrophenol derivatives were detected at 365 nm. The retention times were 13.9 min and 15.1 min and the lowest limits of detection were 10 pmol and 5 pmol for reduced and oxidized glutathione, respectively. This HPLC method was applicable to a variety of woody plant species and different tissues. Total glutathione content in the bark tissue of seven of the woody plant species tested were lower during the period of active plant growth, and higher during the dormant stage. Such change was not observed in grape. Reduced glutathione always represented a high percentage of the total glutathione. All tested tissues of peach plants contained glutathione. The highest and lowest levels were found in the leaves and roots, respectively. Both cherry and sugar pine seeds contained high levels of reduced glutathione.
日本植物生理学会の論文