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Journal of Lipid Research, Vol 37, 2608-2615, Copyright © 1996 by Lipid Research, Inc.
M Schlame, R Haupt, I Wiswedel, WJ Kox and B Rustow
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.
ARTICLES
Identification of short-chain oxidized phosphatidylcholine in human plasma
Department of Anesthesiology and Intensive Therapy, University Hospital Charite, Humboldt University, Berlin, Germany.
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