Phosphatidylcholines hydroperoxide determination

A procedure for determination of lipid hydroperoxides in human plasma is based on kinetic measurement of the CL of luminol (124) with hemin (75a) catalysis . CLD of microperoxidase-catalyzed oxidation of luminol (124) or isoluminol (190) was applied to detection and determination of amino acid hydroperoxides after exposure to UV and y-irradiation A method for determination of hydroperoxides in the phospholipids of cultured cells uses isoluminol (190) and microperoxidase as catalyst " . Simultaneous determination of phosphatidylcholine hydroperoxides and cholesteryl ester hydroperoxides in human serum is carried out by quantitative extraction of the lipids, HPLC separation by column switching and CLD using isoluminol (190) with microperoxidase catalysis . ... 

Miyazawa, T., Suzuki, T., Eujimoto, K. and Yasuda, K. Chemiluminescent simultaneous determination of phosphatidylcholine hydroperoxide and phosphatidylethanolamine hydroperoxide in the liver and brain of the rat. J. Lipid Res. 33, 1051-1058 (1992). 

The hydroperoxides of various phospholipids, such as phosphatidylcholine (PC— OOH, 155), phosphatidylserine (PS—OOH), phosphatidylethanolamine (PE—OOH), phos-phatidylinositol (PI—OOH) and cardiolipin (CL—OOH), can be determined by HPLC-ELD on a polar LC—NH2 column, using as mobile phase a MeOH//-PrOH/40 mM aqueous NaH2P04 mixture (61 30 9 by volume). ELD is by the hanging Hg drop electrode method set at —150 mV vs. SCSE. The relative mobility for the given chromatographic system is shown in equation 68, which is almost a reversal of the order shown in equation 63 for HPTLC. The LOD varies from about 0.5 to 1.0 pmol, depending on the class of phospholipid hydroperoxide . ... 

P Therond, M Couturier, JF Demelier, F Lemonnier. Simultaneous determination of the main molecular species of soybean phosphatidylcholine or phosphatidylethanolamine and their corresponding hydroperoxides obtained by lipoxygenase treatment. Lipids 28 245-249, 1993. 

Chemiluminescence has been used to assess phosphatidylcholine oxidation, and to measure the kinetics of decomposition of hydroperoxides formed during the oxidation of soya phosphatidylcholine. The direct chemiluminescence method correlated well with other methods of determining oxidation status (chemical, UV, HPLC, and microcalorimetry), and it was concluded that chemiluminescence was an ideal method for estimating the oxidation of phosphatidylcholine (and phospholipids in general). Kinetics measurements revealed that... 

The direct separation of phosphohpidhydroperoxides has now been achieved by further advances in HPLC and increased sensitivity by chemiluminescence detection. By reverse-phase HPLC, the hydroperoxides of phosphatidylcholine and phosphatidylethanolamine were separated and determined in stored spray-dried eggs (2.4. 4%), fresh ground meat (0.2%), and raw fish (0.4%). Silica HPLC with post-column chemiluminescence detection has been used to determine phospholipid hydroperoxides in human blood plasma and in the low-density lipoprotein fraction. 

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