Solvent effects in the spin trapping of lipid oxyl radicals

Studies documenting spin trapping of lipid radicals in defined model systems have shown some surprising solvent effects with the spin trap DMPO. In aqueous reactions comparing the reduction of H2O2 and methyl linoleate hydroperoxide (MLOOH) by Fe2+, hydroxyl (HO.) and lipid alkoxyl (LO.) radicals produce identical four-line spectra with line intensities 1:2:2:1. Both types of radicals react with commonly-used HO. scavengers, e.g. with ethanol to produce .C(CH3)HOH and with dimethylsulfoxide (DMSO) to give .CH3. However, DMSO radicals (either .CH3 or .OOCH3) react further with lipids, and when radicals are trapped in these MLOOH systems, multiple adducts are evident. When acetonitrile is added to the aqueous reaction systems in increasing concentrations, .CH2CN radicals resulting rom HO. attack on acetonitrile are evident, even with trace quantities of that solvent. In contrast, little, if any reaction of LO. with acetonitrile occurs, even in 100% acetonitrile. A single four-line signal persists in the lipid systems as long as any water is present, although the relative intensity of the two center lines decreases as solvent-induced changes gradually dissociate the nitrogen and beta-hydrogen splitting constants. Extraction of the aqueous-phase adducts into ethyl acetate shows clearly that the identical four-line spectra in the H2O2 and MLOOH systems arise from different radical species in this study, but the lack of stability of the adducts to phase transfer may limit the use of this technique for routine adduct identification in more complex systems. These results indicate that the four-line 1:2:2:1, aN = aH = 14.9 G spectrum from DMPO cannot automatically be assigned to the HO. adduct in reaction systems where lipid is present, even when the expected spin adducts from ethanol or DMSO appear confirmatory for HO.. Conclusive distinction between HO. and LO. ultimately will require use of 13C-labelled DMPO or HPLC-MS separation and specific identification of adducts when DMPO is used as the spin trap.