linoleic-acid and 2-2--azobis(2-4-dimethylvaleronitrile)

Lipid alkyl radicals generated from polyunsaturated fatty acids via chemical or enzymatic H-abstraction have been a pathologically important target to quantify. In the present study, we established a novel method for the quantification of lipid alkyl radicals via nitroxyl radical spin-trapping. These labile lipid alkyl radicals were converted into nitroxyl radical-lipid alkyl radical adducts using 3-carbamoyl-2,2,5,5-tetramethyl-3-pyrroline-N-oxyl (CmdeltaP) (a partition coefficient between octanol and water is approximately 3) as a spin-trapping agent. The resulting CmdeltaP-lipid alkyl radical adducts were determined by HPLC with postcolumn online thermal decomposition, in which the adducts were degraded into nitroxyl radicals by heating at 100 degrees C for 2 min. The resulting nitroxyl radicals were selectively and sensitively detected by electrochemical detection. With the present method, we, for the first time, determined the lipid alkyl radicals generated from linoleic acid, linolenic acid, and arachidonic acid via soybean lipoxygenase-1 or the radical initiator 2,2'-azobis(2,4-dimethyl-valeronitrile).

Topics: Arachidonic Acid; Azo Compounds; Biochemistry; Chromatography; Chromatography, High Pressure Liquid; Chromatography, Liquid; Cyclic N-Oxides; Electrochemistry; Free Radicals; Hot Temperature; Linoleic Acid; Lipids; Lipoxygenase; Mass Spectrometry; Models, Chemical; Nitriles; Nitrogen Oxides; Octanols; Sensitivity and Specificity; Spin Labels; Spin Trapping; Temperature; Time Factors; Water

Rosemary is commonly used as a spice and a flavoring agent in food processing. Although the antioxidative properties of its extracts have been investigated, there have been few reports on the volatile components of rosemary. We designed a novel antioxidative system which can generate the volatile constituents in the gaseous phase from a rosemary extract and evaluated the gaseous antioxidative activities against both lipid peroxidation and cell death induced by nitrogen dioxide and ultraviolet radiation. The antioxidative effects of the major volatile components on the oxidation of linoleic acid induced by azo compounds were also investigated in a solution. The volatile components in the novel antioxidative system suppressed the Jurkat cell death induced by nitrogen dioxide and the intracellular formation of reactive oxygen species in fibroblast cells induced by ultraviolet radiation. 1,8-Cineole among the volatile components exerted an antioxidative effect against the oxidation of linoleic acid in a solution induced by azo compounds and ultraviolet radiation. These data suggest that the volatile constituents of a rosemary extract had antioxidative properties and that gaseous exposure antioxidant is a promising method for promoting health.

Topics: Antioxidants; Apoptosis; Azo Compounds; Cell Line; Gases; Humans; Kinetics; Linoleic Acid; Methylation; Nitriles; Nitrogen Dioxide; Oxidation-Reduction; Plant Extracts; Rosmarinus; Spectrum Analysis; Ultraviolet Rays

We have reported that the peroxyl radicals derived from methyl eicosapentaenoate (20:5n-3) are more polar than those from methyl linoleate (18:2n-6) since the former peroxyl radicals have at least two molecules of oxygen in a molecule while the latter peroxyl radical has one. This lowers the oxidizability for 20:5n-3 in aqueous Triton X-100 micelles by enhancing the termination reaction rate for peroxyl radicals and by reducing the rate of propagation since there may be more polar peroxyl radicals derived from 20:5n-3 at the surface than within the micelle core. In this study, we measured the effect of three antioxidants, di-tert-butyl-4-methylphenol (BHT), 2,2,5,7,8-pentamethyl-6-chromanol (PMC) and 2-carboxy-2,5,7,8-tetramethyl-6-chromanol (Trolox), on the oxidation of lipids in aqueous micelle. Antioxidants give a clear induction period during oxidation of 18:2n-6 initiated with a water-soluble radical initiator, and its induction length decreases in the order of BHT > PMC > Trolox. This is consistent with the proposed location of three antioxidants: being in the core of micelle, at the surface, or in aqueous phase, respectively. However, BHT does not inhibit the oxidation of 20:5n-3 efficiently, and its rate of oxidation is slower than that observed in the oxidation of 18:2n-6, supporting the idea that polar peroxyl radicals derived from 20:5n-3 are preferentially located at the surface of the micelle. Similar results were obtained when oxidation was initiated with a lipid-soluble radical initiator except antioxidants had lesser effect on the oxidation rate of 20:5n-3.

Topics: Aerobiosis; Antioxidants; Azo Compounds; Butylated Hydroxytoluene; Chromans; Eicosapentaenoic Acid; Free Radicals; In Vitro Techniques; Linoleic Acid; Micelles; Nitriles; Oxidation-Reduction; Water

The aerobic oxidation kinetics of methyl eicosapentaenoate (20:5n-3) and methyl linoleate (18:2n-6) were compared in homogeneous chlorobenzene solution and in Triton X-100 aqueous micelles at 37 degrees C. The rate of disappearance of 20:5n-3 was two times faster than that of 18:2n-6 in chlorobenzene, while the former was five times slower than the latter in aqueous micelles. It was also observed that delta O2 = delta 18:2n-6 and delta O2 = 2 delta 20:5n-3 in aqueous micelles. In the oxidation of a 1:1 mixture of 20:5n-3 and 18:2n-6 in micelles, the rate of disappearance of 20:5n-3 was 3.6 times faster than that of 18:2n-6, and the rate of total substrate disappearance was reduced by a factor of 5 as compared with 18:2n-6 oxidation. These data suggest that the peroxyl radical derived from 20:5n-3 is more polar than that from 18:2n-6, and the former is likely to diffuse from the core to the micelle surface. This lowers the oxidizability for 20:5n-3 in aqueous micelles by enhancing the termination reaction rate for peroxyl radicals and by reducing the rate of propagation since there may be more 20:5n-3 peroxyl radicals at the surface than in the micelle core.

Topics: Amidines; Azo Compounds; Chlorobenzenes; Eicosapentaenoic Acid; Esters; Fatty Acids, Unsaturated; Kinetics; Linoleic Acid; Linoleic Acids; Micelles; Models, Chemical; Nitriles; Octoxynol; Oxidation-Reduction; Peroxides

The authors have developed a kinetic method that allows one to obtain relative reactivity constants for lipophilic antioxidants in free radical systems. Two experimental model systems were developed: (a) a methanolic solution using AMVN as the free radical initiator and linoleic acid as the substrate, and (b) a multilamellar vesicle system composed of dilinoleoylphosphatidylcholine and AAPH as the substrate and the initiator, respectively. The use of these two systems allows researchers not only to determine the intrinsic reactivity of a potential antioxidant, but also to evaluate its potency in a membranous system where the contribution of the physical properties of the antioxidant to the inhibition of lipid peroxidation is important. These results show that all antioxidants tested acted in these systems as free radical scavengers, and they validate the synergism between intrinsic scavenging ability and membrane affinity and/or membrane-modifying physical properties in the inhibition of lipid peroxidation.

Topics: Amidines; Antioxidants; Azo Compounds; Chromans; Chromatography, High Pressure Liquid; Free Radical Scavengers; Free Radicals; Kinetics; Linoleic Acid; Linoleic Acids; Lipid Peroxidation; Liposomes; Mass Spectrometry; Nitriles; Phosphatidylcholines; Piperazines; Pregnatrienes