Autor: |
M Schlame, R Haupt, I Wiswedel, W J Kox, B Rüstow |
Jazyk: |
angličtina |
Rok vydání: |
1996 |
Předmět: |
|
Zdroj: |
Journal of Lipid Research, Vol 37, Iss 12, Pp 2608-2615 (1996) |
Druh dokumentu: |
article |
ISSN: |
0022-2275 |
DOI: |
10.1016/S0022-2275(20)37464-2 |
Popis: |
Oxidized phospholipids have been recognized as potentially important compounds that carry biological activities similar to the platelet-activating factor, but their presence in biological tissue has not been firmly established. We developed a novel technique for the quantitative analysis of phospholipids with oxidized acyl chains. The method involves 1) lipid extraction, 2) chromatographic enrichment of phospholipids with short acyl chains, 3) derivatization with 9-(chloromethyl)anthracene, 4) solid-phase extraction of the derivatives, and 5) reversed-phase HPLC with fluorescence detection. The technique was capable of measuring dicarboxylate-containing phosphatidylcholines (PCs) at the picomole level. The method was suited to monitor the generation of oxidized phospholipids from 1-palmitoyl-2-arachidonoyl-PC in the presence of Fe21/ascorbate. The new procedure was used to isolate lipids from human plasma that were identified as anthracene derivatives of short-chain oxidized PC on the basis of chromatographic enzymatic, and spectroscopic evidence. The plasma concentration, determined with an internal standard (1-palmitoyl-2-suberoyl-PC), was 0.6 +/- 0.2 microM (n = 11). The analytical method did not produce oxidation antifacts in significant amount. We concluded that human blood contains oxidatively fragmented PC in submicromolar concentration. |
Databáze: |
Directory of Open Access Journals |
Externí odkaz: |
|