Effects of α-tocopherol and ascorbyl palmitate on the isomerization and decomposition of methyl linoleate hydroperoxides

In order to study antioxidant action on lipid hydroperoxide decomposition, the effects of α-tocopherol (TOH) and ascorbyl palmitate on the decomposition rate and reaction sequences of 9- and 13-cis,trans methyl linoleate hydroperoxide (cis,trans ML-OOH) decomposition in hexadecane were studied at 40°C. Decomposition of cis,trans ML-OOH as well as the formation and isomeric configuration of methyl linoleate hydroxy and ketodiene compounds were followed by high-performance liquid chromatographic analysis. TOH effectively inhibited the decomposition of ML-OOH. The decomposition rate was two times slower at 0.2 mM and more than 10 times slower at 2 and 20 mM of TOH. Ascorbyl palmitale (0.2, 2, and 20 mM) slightly accelerated the decomposition of ML-OOH. Both compounds had an effect on the reaction sequences of ML-OOH decomposition. At high levels TOH inhibited the isomerization of cis,trans ML-OOH to trans,trans ML-OOH through peroxyl radicals and increased the formation of hydroxy compounds. Further, the majority of the hydroxy and ketodiene compounds formed had a cis,trans configuration, indicating that cis,trans ML-OOH decomposed through alkoxyl radicals without isomerization. These results suggest that when inhibiting the decomposition of hydroperoxides, TOH can act as a hydrogen atom donor to both peroxyl and alkoxyl radicals. In the presence of ascorbyl palmitate, cis,trans ML-OOH decomposed rapidly but without isomerization. In contrast to TOH, the majority of hydroxy compounds were cis,trans, but the ketodiene compounds were trans,trans isomers. This indicates that ascorbyl palmitate reduced cis,trans ML-OOH to the corresponding hydroxy compounds. However, the simultaneous formation of trans,trans ketodiene compounds suggests that ML-OOH decomposition, similar to the control sample, also occurred in these samples. Thus, under these experimental conditions, the reduction of ML-OOH to more stable hydroxy compounds did not occur to an extent significant enough to inhibit the radical chain reactions of ML-OOH decomposition.     

Reaction of α-tocopherol in heated bulk phase in the presence of methyl linoleate (13S)-hydroperoxide or methyl linoleate

α-Tocopherol and methyl (9Z, 11E)-(S)-13-hydroperoxy-9, 11-octadecadienoate (13-MeLOOH) were allowed to stand at 100°C in bulk phase. The products were isolated and identified as methyl 13-hydroxyoctadecadienoate (1), stereoisomers of methyl 9,11,13-octadecatrienoate (2), methyl 13-oxo-9, 11-octadecadienoate (3), epoxy dimers of methyl linoleate with an ether bond (4), a mixture of methyl (E)-12, 13-epoxy-9-(α-tocopheroxy)-10-octadecenoates and methyl (E)-12, 13-epoxy-9-(α-tocopheroxy)-11-(α-tocopheroxy)-9-octadecenoates (5), a mixture of methyl 9-(α-tocopheroxy)-10,12-octadecadienoates and methyl 13-(α-tocopheroxy)-9, 11-octadecadienoates (6), α-tocopherol spirodiene dimer (7), and α-tocopherol trimer (8). α-Tocopherol and 13-MeLOOH were dissolved in methyl myristate, and the thermal decomposition rate and the distributions of reaction products formed from α-tocopherol and 13-MeLOOH were analyzed. α-Tocopherol disappeared during the first 20 min, and the main products of α-tocopherol were 5 and 6 with the accumulation of 1–4 which were the products of 13-MeLOOH. The results indicate that the alkyl and alkoxyl radicals from the thermal decomposition of 13-MeLOOH could be trapped by α-tocopherol to produce 5 and 6. The reaction products of α-tocopherol during the thermal oxidation of methyl linoleate were compounds 6 and 7. Since the radical flux during the autoxidation might be low, the excess α-tocopheroxyl radical reacted with each other to form 7.     

Hydrolysis of methyl α- and β-D-glucopyranosides in the presence of a dealuminated H-Y faujasite

Hydrolysis of methyl α- and β-D-glucopyranosides was performed in the presence of protonic-form zeolites such as a dealuminated Y-faujasite with a Si/Al ratio of 15, at temperatures ranging between 100 and 150°C, and in water as the solvent. The β/α ratio for the hydrolysis reaction rates was found to be equal to 5–6, whereas a ratio of 2–3 was reported in the literature for the homogeneous reaction. The observed higher β/α ratio is proposed to result from the reinforcement of stereoelectronic effects which were shown to apply in reactions taking place on the surface of a solid. Those effects operate in a classical manner on a molecular standpoint, but they are reinforced due to the favorable interaction of oxygen electron lone pairs with the electron-deficient species present on the surface of the solid, protonic species in the case of zeolites. This revised version was published online in August 2006 with corrections to the Cover Date.     

A rigorous kinetic model for β-carotene oxidation in the presence of an antioxidant, α-tocopherol

Oxidation of β-carotene in an inert solvent, n-decane, in the presence of various concentrations of the antioxidant α-tocopherol was studied. The progress of carotene oxidation was suppressed as long as the tocopherol remained in the system. A rigorous kinetic model for carotene oxidation in the presence of an antioxidant was proposed based on a reaction mechanism in which not only the antioxidation but also the co-oxidation and radical-exchange reaction of tocopherol with carotene were incorporated. The model quantitatively described the oxidation behavior of carotene over a wide range of temperatures, oxygen compositions, and initial antioxidant concentrations.     

Preparation of poly(methyl methacrylate) macromonomer by radical polymerization in the presence of methyl α-(bromomethyl)acrylate and copolymerization of the resultant macromonomer

Radical polymerization of methyl methacrylate (MMA) in the presence of methyl -(bromomethyl) acrylate yielded poly-(MMA) bearing the 2-methoxycarbonylallyl end group through chain reaction involving bimol ecular termination. The molecular weight of the resultant polymer was effectively controlled with a small amount of the bromomethylacrylate added; the chain transfer constant was estimated to be 0.9. The poly (MMA) with the unsaturated end group (

Fourier transform infrared spectroscopy evaluation of low density lipoprotein oxidation in the presence of quercetin,catechin, and α-tocopherol

We investigated the changes in human LDL primary and secondary lipid oxidation products and modification of the apolipoprotein B-100 (apoB-100) secondary structures during Cu²⁺-mediated oxidation by FTIR spectroscopy in the presence of catechin, quercetin, and α-tocopherol at physiological concentrations. Catechin- and quercetin-containing samples had slower rates and longer lag phases for conjugated diene hydroperoxide (CD) formation than α-tocopherol-containing samples; however, all antioxidant-treated LDL samples generated similar CD levels (P<0.05). A lower maximum (98.4 nmol/mg LDL protein) of carbonyl compounds was produced in the quercetin- and catechin-treated samples than in α-tocopherol samples. Modification of the apoB-100 secondary structures corresponded closely to the formation of carbonyls and was hampered by the presence of antioxidants. Physiological concentrations of catechin and quercetin offered similar levels of protection against modification by carbonyls of the apoB-100 at advanced stages (carbonyls ∼96.0 nmol/mg LDL protein) but not at the intermediate stages (carbonyls ∼58.0 nmol/mg LDL protein) of LDL oxidation probably owing to differences in the protein-binding mechanisms of catechin and quercetin. Relationships between peroxide formation, carbonyl products, and LDL protein denaturation were shown by the FTIR approach. The FTIR technique provided a simple new tool for a comprehensive evaluation of antioxidant performance in protecting LDL during in vitro oxidation.     

Isomerization of α-pinene oxide in the presence of methyltrioxorhenium(VII)

The catalytic performance of methyltrioxorhenium(VII) (MTO) has been investigated for the first time in the isomerization of α-pinene oxide (PinOx) into campholenic aldehyde (CPA). The high isomerization activity of MTO is coupled with high selectivity to CPA: CPA yield of up to 87% (100% conversion) was obtained by using α,α,α-trifluorotoluene as solvent at 15 °C. Catalyst recycling is possible in a relatively simple fashion by using MTO coupled to an appropriate ionic liquid. The catalytic application of MTO in the isomerization of PinOx versus the integrated epoxidation–isomerization process of the conversion of α-pinene into CPA is discussed.     

Effects of α- and γ-tocopherols on formation of hydroperoxides and two decomposition products from methyl linoleate

The antioxidant effects of α-and γ-tocopherols (at 0, 10, 100, 500, and 1000 ppm) were evaluated in a model system based on the autoxidation of methyl linoleate in bulk for 4 d at 40°C. Samples were collected every 24 h and analyzed for the 9 cis,trans, 9 trans,trans, 13 cis,trans, and 13 trans,trans isomers of hydroperoxide, hydroxy, and ketodiene oxidation products by high-performance liquid chromatography. Results showed that both α- and γ-tocopherols are effective hydrogen donors as evidenced by their abilities to inhibit the formation of hydroperoxides, hydroxy compounds, and ketodienes and the cis,trans to trans,trans isomerization of hydroperoxides. Compared with γ-tocopherol, α-tocopherol was a more efficient antioxidant at very low concentrations (10 ppm) but a less efficient antioxidant at the high concentrations (100–1000 ppm). This paradoxical behavior is explained on the basis of differences in ease of hydrogen donation between the two tocopherol homologs. Although α-tocopherol shows some loss of efficiency with increasing concentration, it is not a prooxidant when compared to the control void of antioxidants.     

Oxidative catabolism of α-tocopherol in rat liver microsomes

The goal of this study was to clarify the mechanism responsible for the catabolism of α-tocopherol. The vitamin, bound to albumin, was incubated with rat liver microsomes and appeared to be broken down. Optimal production of the metabolite was obtained when 1 mg of microsomal protein was incubated with 36 μM of α-tocopherol in the presence of 1.5 mM of NADPH. Chromatographic and mass spectrometric analyses of the metabolite led to the conclusion that it consists of an ω-acid with an opened chroman ring, although we could not perform nuclear magnetic resonance analysis to confirm this. Our data show that α-tocopherol is ω-oxidized to a carboxylic acid and that this process can occur in rat liver microsomes in the presence of NADPH and O₂. The oxidation to the quinone structure appears to be a subsequent event that may be artifactual and/or catalyzed by a microsomal enzyme(s).     

Protection of α-tocopherol in nonpurified and purified fish oil

Menhanden oil was purified by column chromatography to remove minor components. The effect of α-tocopherol (α TOH) (50–500 ppm) on the rate of formation of hydroperoxides in the original menahaden oil and in the purified menhaden triacylglycerol (TAG) fraction was studied at 30°C in the dark. An increase in the initial rate of formation of hydroper-oxides was observed at αTOH concentrations above 100 ppm in both substrates. The original menhaden oil oxidized more rapidly than the purified menhaden, TAG at all antioxidant levels tested, and the presence of minor components in the menhaden oil was found to contribute only to a limited extent to the peroxidizing effect of αTOH. The αTOH did not display any prooxidant activity at either of the concentrations tested when the control oil was the purified menhaden TAG. Addition of ascorbyl palmitate eliminated the initial peroxidizing effect of αTOH, and this emphasizes the participation of the α-toco-pheroxyl radical in the reactions causing an accumulation of hydroperoxides at high concentrations of αTOH. Presented in part at the Annual Meeting of the American Oil Chemists’ Society in San Diego, April 25–28, 2000.     

Antioxidant synergy of α-tocopherol and phospholipids

The prevention of oxidation of a refined sardine oil by α-tocopherol at 0.04%, by several phospholipids [phosphatidylcholine (PC), phosphatidylethanolamine (PE), and cardiolipin (CL)] at 0.5%, as well as by combinations of α-tocopherol with each phospholipid, was investigated. The evolution of the oxidation process during 1 mon at 40±2°C was followed by a series of methods, measuring peroxide value (PV), diene, triene, and polyene index, and absorbance at 430 nm, while α-tocopherol and phospholipid content were being monitoried. Among these indices, PV was found to be the most adequate to follow the process. PC was the most effective individual antioxidant as shown by the PV values obtained at the end of the storage period, which were 54.0, 83.4, 87.9, and 97.7 meq O₂/kg for PC, CL, PE, and α-tocopherol, respectively. The highest synergistic effect was obtained with a mixture of α-tocopherol and PE, and the second and third best by mixtures made with PC and CL, respectively. The corresponding PV values recorded at the end of the period were 27.0, 35.0, and 58.0 meq O₂/kg. The high degree of synergy between PE and tocopherol is probably due to the occurrence of a simultaneous antioxidant mechanism involving Maillard compounds.     

Electrocatalytic carboxylation of aromatic ketones with carbon dioxide in ionic liquid 1-butyl-3-methylimidazoliumtetrafluoborate to α-hydroxy-carboxylic acid methyl ester

A new electrochemical procedure for the electrocatalytic carboxylation of aromatic ketones with carbon dioxide in ionic liquid, 1-butyl-3-methylimidazolium tetrafluoborate (BMIMBF₄), to α-hydroxycarboxylic acid methyl ester was investigated for the first time. The electrochemical behavior of acetophenone in BMIMBF₄ was studied by cyclic voltammetry with a reduction peak at −1.9 V (vs. Ag). The electrolyses experiments were carried out in an undivided cell under mild conditions without any toxic solvents, catalysts and supporting electrolytes, followed by addition of an alkylating agent, affording the α-hydroxycarboxylic acid methyl ester in a moderate yield (62%). The results showed that the yields were strongly affected by various factors: temperature, current density, charge passed, electrode material and substrate concentration. Moreover, the ionic liquid was successfully recycled for this reaction.     

Incorporation of α-tocopherol in marine lipid-based liposomes: in vitro and in vivo studies

Liposomes made from a natural marine lipid extract and containing a high polyunsaturated n−3 fatty lipid ratio were envisaged as oral route vectors and a potential α-tocopherol supplement. The behavior of vesicles obtained by simple filtration and of giant vesicles prepared by electroformation was investigated in gastrointestinal-like conditions. The influence of α-tocopherol incorporation into liposomes was studied on both physical and chemical membrane stability. Propanal, as an oxidation product of n−3 polyunsaturated fatty acids, was quantified by static headspace gas chromatography when α-tocopherol incorporation into liposome ratios ranged from 0.01 to 12 mol%. Best oxidative stability was obtained for liposomes that contained 5 mol% α-tocopherol. Compared to the other formulas, propanal formation was reduced, and time of the oxidation induction phase was longer. Moreover, α-tocopherol induced both liposome structural modifications, evidenced by turbidity, and phospholipid chemical hydrolysis, quantified as the amount of lysophospholipids. This physicochemical liposome instability was even more pronounced in acid storage conditions, i.e., α-tocopherol incorporation into liposome membranes accelerated the structural rearrangements and increased the rate of phospholipid hydrolysis. In particular, giant vesicles incubated at pH 1.5 underwent complex irreversible shape transformations including invaginations. In parallel, the absorption rate of α-tocopherol was measured in lymph-cannulated rats when α-tocopherol was administrated, as liposome suspension or added to sardine oil, through a gastrostomy tube. α-Tocopherol recovery in lymph was increased by almost threefold, following liposome administration. This may be related to phospholipids that should favor α-tocopherol solubilization and to liposome instability in the case of a high amount of α-tocopherol in the membranes. A need to correlate results obtained from in vitro liposome behavior with in vivo lipid absorption was demonstrated by this study.     

Effect of temperature and addition of α-tocopherol on the oxidation of trilinolein model systems

The effects of temperature and addition of α-tocopherol were evaluated in trilinolein model systems through quantification of oxidized TAG monomers, dimers, and polymers following oxidation at different temperatures. Samples of trilinolein without and with 250 and 500 mg/kg α-tocopherol added were stored at 25, 60, and 100°C. Quantification of oxidized monomers, dimers, and polymers by a combination of adsorption and exclusion chromatography provided a useful measurement for studying the evolution of oxidation. Results showed that the amounts of primary oxidation compounds (trilinolein oxidized monomers) that accumulated during the induction period decreased as the temperature increased, indicating that the slope of the initial linear stage of oxidation depended on temperature. The end of the induction period was marked by a sharp increase in the levels of total oxidation compounds, the initiation of polymerization, and the loss of α-tocopherol. Addition of α-tocopherol did not prevent, but rather delayed, formation of trilinolein oxidized monomers and the initiation of polymerization.     

Intermediates and products formed during fatty acid α-oxidation in cucumber (Cucumis sativus)

Fatty acid α-oxidation is an essential metabolic pathway both in plants and in mammals which is still not completely understood. We previously described and purified an α-oxidation enzyme in cucumber which has been used in the present investigation of the α-oxidation reaction mechanism. Free fatty acids, and not the CoA thioesters, were found to undergo α-oxidation in cucumber. 2-Hydroxy- and 2-oxopalmitic acids were identified as palmitic acid α-oxidation intermediates by high-performance liquid chromatography and gas chromatography—mass spectrometry analysis in cucumber subcellular 150,000×g _max pellets obtained by differential centrifugation. Incubation of purified α-oxidation enzyme with [1-¹⁴C]palmitic acid resulted in the formation of both the above-described intermediates and the C _n−1 product, pentadecanal, and ¹⁴CO₂. Besides ¹⁴CO₂, ¹⁴C-formate was identified as an α-oxidation product from [1-¹⁴C]palmitic acid in cucumber subcellular fractions. Fe²⁺ stimulated the ¹⁴CO₂ and ¹⁴C-formate production, and the addition of ascorbate and 2-oxoglutarate together with Fe²⁺ resulted in optimal α-oxidation activities, suggesting a dioxygenase reaction mechanism, as previously shown in mammals. NADPH and, to a lesser extent, NADH stimulated the total ¹⁴C-formate plus ¹⁴CO₂ production but had only slight or no effects on ¹⁴CO₂ production. H₂O₂ showed concentration-dependent inhibitory effects, while FAD had neither effect on ¹⁴CO₂ nor ¹⁴CO₂ plus ¹⁴C-formate production. The results in the present study demonstrate that an α-oxidation enzyme in cucumber is capable of oxidizing palmitic acid via 2-hydroxy- and 2-oxopalmitic acid to produce pentadecanal and CO₂. In contrast to the subcellular 150,000×g _max fraction, the purified α-oxidation enzyme could neither produce formate nor convert ¹⁴C-formate into ¹⁴CO₂, indicating two possible α-oxidation routes in cucumber.     

Regulation of the α-tocopherol transfer protein in mice: Lack of response to dietary vitamin E or oxidative stress

The α-tocopherol transfer protein (TTP) plays an important role in the regulation of plasma α-tocopherol concentrations. We hypothesized that hepatic TTP levels would be modulated by dietary vitamin E supplementation and/or by oxidative stress. Mice were fed either a High E (1150 mg RRR-α-tocopheryl acetate/kg diet) or a Low E (11.5 mg/kg diet) diet for 2 wk. High E increased plasma and liver α-tocopherol concentrations approximately 8- and 40-fold, respectively, compared with Low E-fed mice, whereas hepatic TTP increased approximately 20%. Hepatic TTP concentrations were unaffected by fasting (24 h) in mice fed either diet. To induce oxidative stress, chow-fed mice were exposed for 3 d to environmental tobacco smoke (ETS) for 6 h/d (total suspended particulate, 57.4±1.8 mg/m3). ETS exposure, while resulting in pulmonary and systemic oxidative stress, had no effect on hepatic α-tocopherol concentrations or hepatic TTP. Overall, changes in hepatic TTP concentrations were minimal in response to dietary vitamin E levels or ETS-related oxidative stress. Thus, hepatic TTP concentrations may be at sufficient levels such that they are unaffected by either modulations of dietary vitamin E or by the conditions of environmentally related oxidative stress used in the present studies. The first and second investigators contributed equally to this work.     

Detection and characterization of molybdenum oxides formed during the initial stages of cobalt–molybdenum electrodeposition

The initial stages of cobalt–molybdenum electrodeposition on a vitreous carbon electrode were studied to obtain information about the mechanism of cobalt–molybdenum induced codeposition. Solutions containing cobalt sulphate, sodium molybdate and sodium citrate at pH 6.6 were used. A first step in the mechanism of alloy deposition is proposed. This step takes into account the formation of molybdenum(IV) oxides over which Co–Mo alloy may be only deposited if sufficient potential is applied. Co–Mo electrodeposition occurs through an early stage involving low reduction current, related to the formation of molybdenum oxides, followed by a later stage in which the reduction current suddenly increases, corresponding to alloy codeposition. When a low potential is applied, a continuous coloured molybdenum oxide film is formed on the electrode and Co–Mo is not deposited. To induce the alloy deposition on the oxide film it is necessary to apply more negative potentials than a threshold value, which depends on the composition of the electrolytic bath. By increasing molybdate concentration in solution, the threshold potential shifts to more negative values. Intermediate molybdenum oxides were characterized using scanning electron microscopy (SEM), compositional analysis, Raman measurements and Auger and X-ray photoelectron spectroscopies.     

Electrochemical deposition of zinc–polystyrene composites in the presence of surfactants

The electrochemical codeposition of polystyrene particles and zinc on a rotating cylinder electrode was investigated. Rheological measurements indicate strong aggregation of the PS particles in the zinc deposition electrolyte. Addition of cetylpyridinium chloride, a cationic surfactant, prevents aggregation and enhances polystyrene codeposition. Other surfactants also increase suspension stability, but diminish polystyrene codeposition, irrespective of their charge. Hence, the surfactant charge does not affect polystyrene codeposition. The variation of polystyrene incorporation with the amount of suspended polystyrene, current density and electrode rotation speed signifies that polystyrene codeposition with zinc is determined by the competition between particle removal forces and particle adhesion forces at the cathode surface. The effect of the surfactants can be related to changes in surface roughness of zinc due to surfactant adsorbed on the electrode. Cetylpyridinium chloride behaves differently from the other surfactants, because it is reduced at the cathode.