Using a modified ferrous oxidation-xylenol orange (FOX) assay for detection of lipid hydroperoxides in plant tissue

被引:131
作者
DeLong, JM
Prange, RK
Hodges, DM
Forney, CF
Bishop, MC
Quilliam, M
机构
[1] Agr & Agri Food Canada, Atlantic Food & Hort Res Ctr, Kentville, NS B4N 1J5, Canada
[2] Natl Res Council Canada, Inst Marine Biosci, Halifax, NS B3L 3V2, Canada
关键词
fatty acid oxidation; ferrous oxidation - xylenol orange (FOX) assay; lipid hydroperoxides; oxidative stress;
D O I
10.1021/jf0106695
中图分类号
S [农业科学];
学科分类号
09 ;
摘要
The ferrous oxidation-xylenol orange (FOX) assay was adapted for quantifying lipid hydroperoxides (LOOHs) in plant extracts. Excised pieces of several fruit and vegetable species were exposed to 83 kJ m(-2) day(-1) of biologically effective ultraviolet B irradiance (UV-B-BE) for 10-12 days to induce cellular oxidation. The LOCH and thiobarbituric acid reactive substance (TBARS) concentrations of these plant tissues were assessed with the FOX and iodometric assays for the former and a modified TBARS assay for the latter. There was generally good agreement between the FOX and iodometric methods both prior to and following the UV exposure. However, the iodometric assay appeared to have some difficulty in consistently quantifying lower LOOH levels (<11 muM), whereas the FOX assay measured LOCH concentrations as low as 5 muM. All tissues exhibited UV-induced increases in TBARS, indicating a marked degree of cellular oxidation in the exposed tissue segments. Compared with the iodometric assay, the FOX method consistently generated less variable LOOH values. The presence of authentic linoleic acid-OOHs in spiked avocado and spinach samples (11 muM) was identified with liquid chromatography-mass spectrometry techniques, which validated corresponding FOX assay results. The FOX method is inexpensive, is not sensitive to ambient O-2 or light levels, and can rapidly generate LOOH measurements. The physiological value of the FOX assay resides in its ability to measure initial rather than more advanced fatty acid oxidation; hence, early membrane-associated stress events in plant tissue can be detected.
引用
收藏
页码:248 / 254
页数:7
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