Phospholipids hydroperoxide determination

Miyazawa T, Fujimoto K, Suzuki T, Yasuda K. Determination of phospholipid hydroperoxides using luminol chemiluminescence high-performance liquid chromatography. Oxygen Radicals in Biological Systems, Pt. C 233, 324-332, 1994. 

The CLD methods for HPLC using isoluminol (190) with microperoxidase catalysis, for determination of lipid hydroperoxides in clinical fluids, have been reviewed. Determination of phospholipids hydroperoxides by luminol (124) CL has been reviewed . A fast RP-HPLC method (retention times 1 to 2 min) for determination of hydroperoxides and other peroxide compounds includes UVD, which is not always effective, and CLD, attained on injection of luminol (124), the CL reagent (Scheme 3), hemin (75a), a catalyst, and NaOH to raise the pH of the solution. A FLD cell may act as CLD cell if the excitation source is turned off. The selectivity of CLD is of advantage over UVD in industrial analysis thus, for example, UVD of a sample from a phenol production line based on cumene oxidation (equation 13) shows peaks for cumyl hydroperoxide (27), unreacted cumene, cumyl alcohol and acetophenone, whereas CLD shows only the 27 peak. The... 

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 . ... 

Asai et al. (1999) determined that phospholipid hydroperoxides (PLOOH) are key products for oxidative injury in membranous phospholipid layers in the plasma, red blood cells (RBC), and liver of mice. The formation and accumulation of PLOOH have been confirmed in several cellular disorders, various diseases, and in aging. A lower PLOOH level was found in RBC of the spice-extract-fed mice (65 to 74% of the nonsupplemented control mice). The liver lipid peroxidizability induced with Fe2+/ascorbic acid was effectively suppressed in mice by dietary supplementation with the turmeric and capsicum extracts. Although no difference in the plasma lipids was observed, the liver triacylglycerol concentration of the turmeric-extract-fed mice was markedly reduced to half of the level in the control mice. These findings suggest that these spice extracts could act antioxidatively in vivo by food supplementation, and that the turmeric extract has the ability to prevent the deposition of triacylglycerols in the liver. 

The oxidation products of plasma and lipoprotein phospholipids have been the subject of extensive study by chromatographic and mass spectrometric methods. Earlier, workers had reported the simultaneous determination of phospholipid hydroperoxides and cholesteryl ester hydroperoxides in human plasma by HPLC with chemiluminescence monitoring. Lipid hydroperoxides were quantitatively extracted from human plasma with a mixture of -hexane and... 

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. 

Terao, J. and Matsushita, S. Application of high-performance liquid chromatography for the determination of phospholipid hydroperoxides in foods and biological systems. Free Radical Biol. Med. 3, 345-348 (1987). 

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 . ... 

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... 

---