Analysis of F2-isoprostanes as indicators of non-enzymatic lipid peroxidation in vivo by gas chromatography-mass spectrometry: development of a solid-phase extraction procedure
| Autor: | A.I. Mallet, S. Barrow, Jaffar Nourooz-Zadeh, Erik Änggård, Nitin K Gopaul |
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| Rok vydání: | 1995 |
| Předmět: |
Detection limit
Chemical ionization Chromatography Esterification Chemistry General Chemistry Silanes Mass spectrometry Deuterium Dinoprost Gas Chromatography-Mass Spectrometry Cyclooxygenase pathway Lipoproteins LDL F2-Isoprostanes Humans Solid phase extraction Gas chromatography Lipid Peroxidation Gas chromatography–mass spectrometry |
| Zdroj: | Journal of chromatography. B, Biomedical applications. 667(2) |
| ISSN: | 1572-6495 |
| Popis: | Recently, it has been reported that a series of prostaglandin F2-like compounds (F2-isoprostanes) are produced in vivo during peroxidation of arachidonic acid by a mechanism independent of the cyclooxygenase pathway. Of these, 8-epi-PGF2 alpha is shown to be a potent vasoconstrictor. We describe an improved method for analysing F2-isoprostanes in biological fluids. The method involves solid-phase extraction on an octadecylsilane (C18) and an aminopropyl (NH2) cartridge. After conversion to pentafluorobenzyl ester and trimethylsilyl ether derivatives, F2-isoprostanes are analysed by negative-ion chemical ionization mass spectrometry using tetradeuterated PGF2 alpha as the internal standard. The limit of detection of the assay was 10 pg/ml, with a coefficient of variation ranging from 9.4 to 15.1%. Analysis of plasma samples from healthy volunteers (n = 7) revealed no quantifiable levels of free (unesterified) 8-epi-PGF 2 alpha. However, the plasma samples contained 58 to 166 pg/ml of 8-epi-PGF2 alpha when analyzed for the total (sum of free and esterified) F2-isoprostanes. The main advantages of the method lie in the improved recovery, gas chromatographic separation and speed compared to existing techniques. |
| Databáze: | OpenAIRE |
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