Abstract
A new method for the detection of phospholipid hydroperoxides in the low nanomolar range has been developed by using a high performance liquid chromatography system combined with a post-column reaction. The detection is based on the oxidation of Fe(II) to Fe(III) by hydroperoxides and the subsequent formation of an ironthiocyanate complex, which can be determined spectrophotometrically at 505 nm. By this method it is possible to separate and quantitate phosphatidylethanolamine hydroperoxide and phosphatidylcholine hydroperoxide formed in erythrocyte ghost membranes during photo-oxidation.
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Abbreviations
- EDTA:
-
ethylenediaminetetraacetic acid
- HPLC:
-
high performance liquid chromatography
- MDA:
-
malondialdehyde
- PC:
-
phosphatidylcholine
- PE:
-
phosphatidylethanolamine
- PC-OOH:
-
phosphatidylcholine hydroperoxide
- PE-OOH:
-
phosphatidylethanolamine hydroperoxide
- TBA:
-
thiobarbituric acid
- TLC:
-
thin-layer chromatography
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Mullertz, A., Schmedes, A. & Hølmer, G. Separation and detection of phospholipid hydroperoxides in the low nanomolar range by a high performance liquid chromatography/irothiocyanate assay. Lipids 25, 415–418 (1990). https://doi.org/10.1007/BF02537987
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DOI: https://doi.org/10.1007/BF02537987