Quantitative determination of hydroxy fatty acids as an indicator of in vivo lipid peroxidation: Gas chromatography-mass spectrometry methods
| Autor: | Frederik J.G.M. van Kuijk, Robert J. Stephens, David W. Thomas, Edward A. Dratz |
|---|---|
| Rok vydání: | 1991 |
| Předmět: |
Male
Lipid Peroxides Trimethylsilyl Compounds Biophysics Mass spectrometry Biochemistry Gas Chromatography-Mass Spectrometry Lipid peroxidation chemistry.chemical_compound Eicosanoic Acids Animals Organic chemistry Molecular Biology chemistry.chemical_classification Chemical ionization Chromatography Esterification Autoxidation Chemistry Fatty Acids Rats Inbred Strains Cell Biology Rats Fluorobenzenes Hydrogenation Lipid Peroxidation Gas chromatography Gas chromatography–mass spectrometry Hydroxy Acids Oxidation-Reduction Saponification Polyunsaturated fatty acid |
| Zdroj: | Analytical Biochemistry. 198:104-111 |
| ISSN: | 0003-2697 |
| Popis: | A new method has been developed for the quantitation of lipid peroxidation products by gas chromatography-mass spectrometry. An important advantage over existing gas chromatography-mass spectrometry methods is the elimination of autoxidation during sample preparation. The sensitivity is sufficient to permit measurement of lipid peroxidation products under normal physiological conditions on as little as 1 mg of tissue. Lipids from whole tissue samples or cell preparations are reduced by catalytic hydrogenation during extraction. The hydrogenation stabilizes the compounds by saturating the double bonds and reducing the hydroperoxides to hydroxy derivatives. The saturated lipids are then saponified and the resulting fatty acids are converted to pentafluorobenzyl esters. Hydroxy fatty acids are further converted to trimethylsilyl ether derivatives. Quantitation is accomplished by negative ion chemical ionization gas chromatography-mass spectrometry, using deuterated internal standards. Specific products from polyunsaturated fatty acids can be quantitated, and the method differentiates between products produced by free-radical and photooxidation mechanisms. Increased levels of lipid peroxidation products, above normal physiological levels, that result from prooxidant conditions, such as exposure of animals to carbon tetrachloride, can be measured. |
| Databáze: | OpenAIRE |
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