Separation and identification of phospholipid peroxidation products.
The molecular species in mixtures of phospholipid hydroperoxides are difficult to separate and identify by typical chromatographic and mass spectrometric techniques. As reported by Havrilla and coworkers, silver ion coordination ion-spray mass spectrometry (CIS-MS) has proven to be a powerful technique for the identification of mixtures of hydroperoxides. This ionization technique, which involves the formation of Ag+ adducts of the hydroperoxides, provides valuable, unambiguous structural information about the hydroperoxides. Herein, we report a method for the analysis and identification of phospholipid hydroperoxides using CIS-MS. We also report an improved method for the separation of phospholipid hydroperoxides by reversed-phase high-performance liquid chromatography (RP-HPLC), which, for the first time, separates some of the hydroperoxide isomers. CIS-MS can be coupled with this RP-HPLC method by the addition of AgBF4 to the mobile phase or to the HPLC effluent postcolumn, thus allowing powerful HPLC-MS techniques to be used to identify complex mixtures of phospholipid hydroperoxides.
Milne
G L
GL
Department of Chemistry and Center in Molecular Toxicology, Vanderbilt University, Nashville, Tennessee 37235, USA.
Porter
N A
NA
D016718:Arachidonic Acid / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D019787:Linoleic Acid / Q000032:analysis; D013058:Mass Spectrometry / Q000379:methods*; D010545:Peroxides / Q000032:analysis* / Q000737:chemistry; D010713:Phosphatidylcholines / Q000032:analysis / Q000737:chemistry; D010743:Phospholipids / Q000032:analysis* / Q000737:chemistry; D014466:Ultraviolet Rays
C462319:1-strearoyl-2-arachidonyl-sn-glycero-3-phosphatidylcholine; D010545:Peroxides; D010713:Phosphatidylcholines; D010743:Phospholipids; D016718:Arachidonic Acid; D019787:Linoleic Acid