Measurement of protein and lipid hydroperoxides in biological systems by the ferric-xylenol orange method

Craig A. Gay*, Janusz M. Gebicki

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

    159 Citations (Scopus)

    Abstract

    Methods were developed for the separation and measurement of lipid and protein hydroperoxides, which can be used for biological materials. Lipids were extracted with methanol:chloroform and their hydroperoxides measured in solutions of methanol and chloroform containing 110mM perchloric acid, xylenol orange, and ferrous iron. Proteins were isolated by precipitation with 0.2M perchloric acid. The precipitates were redissolved in 6M guanidine hydrochloride and washed with chloroform, and the hydroperoxides were measured in the presence of perchloric acid, xylenol orange, and ferrous iron. Optimum conditions for hydroperoxide measurements were established and the assays were applied to oxidized human blood serum and to cultured cells.

    Original languageEnglish
    Pages (from-to)29-35
    Number of pages7
    JournalAnalytical Biochemistry
    Volume315
    Issue number1
    DOIs
    Publication statusPublished - 1 Apr 2003

    Keywords

    • hydroperoxide
    • lipid peroxide
    • protein peroxide
    • biological hydroperoxides
    • blood serum
    • cells
    • xylenol orange

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