Simultaneous determination of the main molecular species of soybean phosphatidylcholine or phosphatidylethanolamine and their corresponding hydroperoxides obtained by lipoxygenase treatment

Lipids
P ThérondF Lemonnier

Abstract

A method for the simultaneous determination of the main molecular species of soybean phosphatidylcholine or phosphatidylethanolamine and their corresponding hydroperoxides is described. Hydroperoxides were formed by incubation of phospholipids with lipoxygenase at pH 9.2. Silicic acid column chromatography (silica Sep-Pak column) was used to separate the phospholipids into phosphatidylcholine and phosphatidylethanolamine. A single C-18 reverse-phase column was employed to separate the main molecular species of soybean phosphatidylcholine or phosphatidylethanolamine and their hydroperoxides by high-performance liquid chromatography. The mobile phase consisted of 5% 10 mM ammonium acetate at pH 5 and 95% methanol. The molecular species of phosphatidylcholine and phosphatidylethanolamine were detected at 205 nm; the eluate was mixed with a chemiluminescence reagent (isoluminol and microperoxidase) and monitored by fluorometry. Under the experimental conditions used, three individual molecular species of both soybean phosphatidylethanolamine and phosphatidylcholine (18:3/18:2, 18:2/18:2 and 16:0/18:2), together with their corresponding hydroperoxides, were identified and quantitated.

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Citations

Mar 31, 2004·Canadian Journal of Physiology and Pharmacology·H VitracM Gardès-Albert
Jul 2, 2008·Chemistry and Physics of Lipids·Yara RhayemDominique Bonnefont-Rousselot
May 7, 2011·Journal of Pineal Research·Catherine MarchettiDominique Bonnefont-Rousselot
Apr 20, 2010·Chemistry and Physics of Lipids·Nazha Sidahmed-AdrarFabrice Collin
Jul 22, 2008·Journal of Lipid Research·Daigo IbusukiTeruo Miyazawa
Mar 12, 1999·Arteriosclerosis, Thrombosis, and Vascular Biology·L ChancharmeM J Chapman
Apr 30, 2011·Arteriosclerosis, Thrombosis, and Vascular Biology·Natacha BellangerMaryse Guerin
Jun 28, 2005·Journal of Pineal Research·J MekhloufiM Gardès-Albert
Jul 8, 2009·Radiation Research·Nazha Sid Ahmed-AdrarMonique Gardès-Albert
Dec 12, 2002·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Anatol KontushM John Chapman
Jul 20, 1999·The Journal of Biological Chemistry·A B FisherS I Feinstein

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