Lipid radicals: properties and detection by spin trapping.
Unsaturated lipids are rapidly oxidized to toxic products such as lipid hydroperoxides, especially when transition metals such as iron or copper are present. In a Fenton-type reaction Fe2+ converts lipid hydroperoxides to the very short-lived lipid alkoxyl radicals. The reaction was started upon the addition of Fe2+ to an aqueous linoleic acid hydroperoxide (LOOH) emulsion and the spin trap in the absence of oxygen. Even when high concentrations of spin traps were added to the incubation mixture, only secondary radical adducts were detected, probably due to the rapid re-arrangement of the primary alkoxyl radicals. With the commercially available nitroso spin trap MNP we observed a slightly immobilized ESR spectrum with only one hydrogen splitting, indicating the trapping of a methinyl fragment of a lipid radical. With DMPO or 5-diethoxyphosphoryl-5-methyl-1-pyrroline N-oxide (DEPMPO) adducts were detected with carbon-centered lipid radical, with acyl radical, and with the hydroxyl radical. We also synthesized lipophilic derivatives of the spin trap DEPMPO in order to detect lipid radical species generated in the lipid phase. With all spin traps studied a lipid-derived carbon-centered radical was obtained in the anaerobic incubation system Fe2+/LOOH indicating the trapping of a lipid radical, possibly generated as a secondary reaction product of the primary lipid alkoxyl radical formed. Under aerobic conditions an SOD-insensitive oxygen-centered radical adduct was formed with DEPMPO and its lipophilic derivatives. The observed ESR parameters were similar to those of alkoxyl radical adducts, which were independently synthesized in model experiments using Fe3+-catalyzed nucleophilic addition of methanol or t-butanol to the respective spin trap.
Stolze
K
K
Institute of Pharmacology and Toxicology, Veterinary University of Vienna, Austria. Hans.Nohl@vu-wien.ac.at
Udilova
N
N
Nohl
H
H
D002244:Carbon / Q000737:chemistry; D003497:Cyclic N-Oxides / Q000494:pharmacology; D007501:Iron / Q000737:chemistry / Q000494:pharmacology; D008041:Linoleic Acids / Q000494:pharmacology; D008054:Lipid Peroxides / Q000494:pharmacology; D008055:Lipids / Q000737:chemistry*; D008956:Models, Chemical; D010100:Oxygen / Q000737:chemistry; D013113:Spin Labels; D019097:Spin Trapping / Q000379:methods*; D013482:Superoxide Dismutase / Q000378:metabolism
D003497:Cyclic N-Oxides; D008041:Linoleic Acids; D008054:Lipid Peroxides; D008055:Lipids; D013113:Spin Labels; C098505:5-diethoxyphosphoryl-5-methyl-1-pyrroline N-oxide; C008285:linoleic acid hydroperoxide; D002244:Carbon; D007501:Iron; D013482:Superoxide Dismutase; D010100:Oxygen