Photometric evaluation of lipid peroxidation products in human plasma and copper oxidized low density lipoproteins: correlation of different oxidation parameters

Abstract

Despite considerable methodological advances, ideal parameters relating to the in vitro assessment of lipoprotein oxidizability are lacking. In this study, some of the more common parameters of lipid peroxidation were measured in 30 plasma samples. The following parameters were determined: conjugated dienes (method 1), reaction of lipid hydroperoxides with a methylene blue derivative (method 2), oxidation of iodide to triiodide (method 3) and an iodometric assay based on the same chemistry but modified to correct for unspecific interferences (method 4). α-Tocopherol in plasma was assayed by high-pressure liquid chromatography. In addition, LDL was isolated from plasma and the susceptibility of individual LDL preparations towards copper-initiated oxidation was characterized. The amount of lipid hydroperoxides found in fresh plasma samples obtained from apparently healthy humans was dependent on the method used for the assessment. Lipid hydroperoxides measured by method 2 were: 8.6 ± 5.8 μM, by method 3: 5.8 ± 1.9 μM and by method 4: 4.2 ± 2.7 μM. Mean values of conjugated dienes (method 1) were 84.6 ± 20.9 μM; The content of α-tocopherol in plasma was 23.6 ± 3.9 μM. Despite the differences in absolute values, a statistically significant correlation was found between values obtained by methods 1, 2 and 3, but not by method 4. An inverse relationship has been observed between the lipid hydroperoxide content in plasma obtained with method 4 and two parameters of LDL oxidation (diene concentration, rate of diene formation) but not with the lag time. Our data suggest that — among the photometric methods evaluated — method 4 might be the most specific for the measurement of plasma (and lipoprotein associated) lipid hydroperoxides.