Tajaddini-Sarmaidi, Javad;
(1997)
Analysis of plasma hydroperoxides: Methodological aspects and levels in health and diseases.
Doctoral thesis (Ph.D.), University College London (United Kingdom).
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Abstract
Increased lipid peroxidation is thought to play an important role in the pathogenesis of atherosclerosis. However, such elevation in lipid peroxidation has been difficult to assess because specific and reproducible methods for the measurement of products of lipid peroxidation have been unavailable. The aim of this thesis is to show the development of a simple and reliable method to measure of plasma hydroperoxides, initial products formed during peroxidation of unsaturated lipids (e.g. fatty acids and cholesterol). Using this as an index of oxidative injury in vivo, I have shown that it feasible to measure plasma hydroperoxides using Ferrous Ion Oxidation in Xylenol Orange (FOX) version 2. The assay is based upon the oxidation of ferrous to ferric ions by hydroperoxides under acidic conditions. The generated ferric ion is complexed by the ferric ion indicator xylenol orange yielding a blue-purple complex with an absorbance maximum at 550-600 nm. Authentic plasma hydroperoxides are determined by a strategy in which the specific hydroperoxide reductant, triphenylphosphine, is used to discriminate between the background signal generated by ferric ions present in plasma and that generated by hydroperoxide in plasma. The method is specific and sensitive; detection limit [less-than]1.0 [mu]M. Inter- and intra assay coefficients of variation are ([less-than]5[percent]). Intra laboratory coefficient of variation is [less-than]10[percent]. Using this assay, I have shown that plasma hydroperoxides were elevated in individuals with non-insulin-dependent diabetes mellitus (NIDDM) (9.4 [plus-minus] 3.5 [mu]M;(n=110) compared to those in healthy controls (3.9 [plus-minus] 2.2 [mu]M; n=60). Relationships between measures of plasma oxidation, metabolic control and selected complication in NIDDM and location of plasma hydroperoxides are discussed with relevance to atherosclerosis.
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D. |
| Title: | Analysis of plasma hydroperoxides: Methodological aspects and levels in health and diseases |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| Additional information: | Thesis digitised by ProQuest. |
| Keywords: | (UMI)AAI10017369; Health and environmental sciences |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10101842 |
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