Abstract
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 ionspray 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.
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Abbreviations
- BHT:
-
butylated hydroxytoluene
- CD:
-
circular dichroism
- CE:
-
Cotton effect
- CID:
-
collision-induced dissociation
- CIS-MS:
-
coordination ion-spray mass spectrometry
- DLI:
-
direct liquid infusion
- ESI-MS:
-
electrospray MS
- FAB-MS:
-
fast atom bombardment MS
- HETE:
-
hydroxyeicosatetraenoic acid
- HODE:
-
hydroxyoctadecadienoic acid
- HPETE:
-
hydroperoxyeicosatetraenoic acid
- HPLC:
-
high-pressure liquid chromatography
- HPODE:
-
hydroperoxyoctadecadienoic acid
- LC:
-
liquid chromatography
- LDL:
-
low density lipoproteins
- MS:
-
mass spectrometry
- methyl-9-HETE:
-
methyl 9-hydroxy-5(Z),8(E), 11(Z)-eicosatetraenoate
- MS/MS:
-
tandem MS
- NMR:
-
nuclear magnetic resonance
- ODS:
-
octadecylsilane
- PC:
-
phosphatidylcholine
- PLPC:
-
1-palmitoyl-2-linoleoyl-sn-glycero-3-phosphatidylcholine
- PLPC-OH:
-
1-palmitoyl-2-linoleoyl-sn-glycero-3-phosphatidycholine hydroperoxide
- PMC:
-
pentamethylchromanol
- PPh3 :
-
triphenylphosphine
- RP:
-
reversed phase
- SAPC:
-
1-stearoyl-2-arachidonyl-sn-glycero-3-phosphatidylcholine
- SRM:
-
selected reaction monitoring
- TLC:
-
thin-layer chromatography
- UV:
-
ultraviolet
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Milne, G.L., Porter, N.A. Separation and identification of phospholipid peroxidation products. Lipids 36, 1265–1275 (2001). https://doi.org/10.1007/s11745-001-0841-2
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DOI: https://doi.org/10.1007/s11745-001-0841-2