Suitability of saturated aldehydes as lipid oxidation markers in washed turkey meat.

The aim of this study was to evaluate the suitability of saturated aldehydes as lipid oxidation markers in washed turkey muscle, by means of headspace solid phase microextraction-gas chromatography (HS-SPME-GC); the results were compared with the widely used thiobarbituric acid-reactive substances (TBARs) method. Changes in TBARs, propanal and hexanal concentrations were determined over time in a model system consisting of turkey muscle washed with a sodium phosphate buffer (pH 5.6). To stop oxidation from occurring during analysis, an antioxidant mixture (EDTA, trolox and propyl gallate) was added immediately before analyses. After antioxidant addition, propanal and TBARs concentrations did not increase during 8h of further storage, while an unexpected decrease in hexanal was observed. To determine if aldehydes were interacting with washed turkey muscle, hexanal and propanal were added to either phosphate buffer or washed muscle and concentrations were monitored for 24h. Neither propanal nor hexanal decreased in the phosphate buffer over time, but the headspace concentration of propanal and hexanal in washed turkey muscle were markedly lower (76% and 96%, respectively) at time zero and continued to decreased up to 24h of storage. Because of this decrease in headspace aldehyde concentrations, TBARs were found to be a more sensitive and accurate marker of oxidative deterioration in washed turkey muscle.

Effect of antioxidants on stabilization of meat products fortified with n-3 fatty acids.

The effects of an n-3 oil emulsion, with and without added antioxidants, on lipid oxidation in n-3 polyunsaturated fatty acid (PUFA)-fortified meat products were studied. An emulsion of n-3 PUFAs was prepared (25% algal oil, 2.5% whey protein isolates, 10mM sodium citrate, 0.2% potassium sorbate, 500ppm of 70% mixed tocopherols, 100µM EDTA, pH 3, pasteurized at 75°C for 30min) and incorporated into fresh ground turkey, and fresh pork sausage (20% fat) to achieve a concentration of 500mg n-3 PUFA/110g meat. An antioxidant combination containing rosemary (0.2% w/w; radical quencher), citrate (0.5% w/w; sequestrant) and erythorbate (1g/kg product; reductant) was prepared and incorporated into ground turkey patties (5cm dia, 1.5cm thick) or fresh pork sausages (5cm dia, 1.5cm thick). Meat products were stored at 4°C or -18°C and analyzed for color (L*, a*, b* values), lipid oxidation (TBARS and lipid hydroperoxides) and n-3 PUFA profile. a* Values of refrigerated ground turkey patties decreased with storage, and an antioxidant combination effect was observed after 4 days (P<0.05). For fresh pork sausages at 4°C, control+antioxidant (CON+ANTI), and n-3+antioxidant (n-3+ANTI) groups showed greater a* values than controls (CON) indicating that the antioxidant combination stabilized meat color. TBARS and lipid hydroperoxides of both n-3 PUFA-enhanced meat products increased with storage (P<0.05); there were no significant changes in TBARS or lipid hydroperoxides for treatments containing the antioxidant combination (P<0.05). The actual level of n-3 PUFA incorporation in both meat products was greater than 87%; n-3 PUFA concentrations did not change within any treatment during storage (P>0.05). These results provide support for including antioxidant protection in n-3 PUFA fortified meat products.

The effects of antioxidant combinations on color and lipid oxidation in n-3 oil fortified ground beef patties.

This study was carried out to determine an effective combination of chelators, reductants and free radical scavengers for enhancing color stability and minimizing lipid oxidation in muscle foods fortified with n-3 fatty acids. Chelators (sodium tripolyphosphate, STPP; sodium citrate, CIT), reductants (sodium erythorbate, ERY) and radical scavengers (butylhydroxyanisole, BHA; mixed tocopherols from two different sources, 30 or 95TOC; rosemary extract, ROSE) were incorporated in various combinations into ground beef (15% fat) with or without n-3 oil fortification (n=8). Individually, STPP and CIT had no significant effect on a* values except day 4, but showed higher a* values when combined with ERY (STPP+ERY and CIT+ERY) (P<0.05). CIT had lower hue angle values than STPP on days 4 and 6, but CIT+ERY showed more discoloration than STPP+ERY (P<0.05). CIT+ERY showed less lipid oxidation than CIT alone (P<0.05), whereas there was no difference between STPP and STPP+ERY. CIT+ERY+ROSE demonstrated higher a* values than CIT+ERY+95TOC on days 4 and 6 (P<0.05); there was no difference between ROSE and 95TOC groups when n-3 oil was incorporated into ground beef patties (P>0.05). The combination of ROSE and ERY appeared to be effective in slowing the decline of a* values. All antioxidant combinations were effective at delaying lipid oxidation when compared to CON or n-3. A combination of CIT, ERY and ROSE was most effective for stabilizing color and delaying lipid oxidation.

Comparison of low-density lipoprotein modification by myeloperoxidase-derived hypochlorous and hypobromous acids.

Myeloperoxidase (MPO), a heme enzyme secreted by activated phagocytes, catalyzes the oxidation of halides to hypohalous acids. At plasma concentrations of halides, hypochlorous acid (HOCl) is the major strong oxidant produced. In contrast, the related enzyme eosinophil peroxidase preferentially generates hypobromous acid (HOBr). Since reagent and MPO-derived HOCl converts low-density lipoprotein (LDL) to a potentially atherogenic form, we investigated the effects of HOBr on LDL modification. Compared to HOCl, HOBr caused 2-3-fold greater oxidation of tryptophan and cysteine residues of the protein moiety (apoB) of LDL and 4-fold greater formation of fatty acid halohydrins from the lipids in LDL. In contrast, HOBr was 2-fold less reactive than HOCl with lysine residues and caused little formation of N-bromamines. Nevertheless, HOBr caused an equivalent increase in the relative electrophoretic mobility of LDL as HOCl, which was not reversed upon subsequent incubation with ascorbate, in contrast to the shift in mobility caused by HOCl. Similar apoB modifications were observed with HOBr generated by MPO/H(2)O(2)/Br(-). In the presence of equivalent concentrations of Cl(-) and Br(-), modifications of LDL by MPO resembled those seen in the presence of Br(-) alone. Interestingly, even at physiological concentrations of the two halides (100 mM Cl(-), 100 microM Br(-)), MPO utilized a portion of the Br(-) to oxidize apoB cysteine residues. MPO also utilized the pseudohalide thiocyanate to oxidize apoB cysteine residues. Our data show that even though HOBr has different reactivities than HOCl with apoB, it is able to alter the charge of LDL, converting it into a potentially atherogenic particle.

The role of phenolics, conjugated linoleic acid, carnosine, and pyrroloquinoline quinone as nonessential dietary antioxidants.

Oxidative reactions have been implicated in the development of numerous diseases including atherosclerosis and cancer. Oxidation of lipids, proteins, and nucleic acids can result in loss of membrane integrity and function, inactivation of enzymes, modification of lipoproteins, and chemical alteration of DNA. Active oxygen species, transition metals, reducing agents, and enzymes such as lipoxygenase are all involved in the catalysis of oxidative reactions. Since lipid oxidation catalysts and active oxygen species are ubiquitous to all biological systems and since lipid oxidation products can enter the body via oxidized foods, numerous endogenous antioxidant systems have been developed. Endogenous antioxidant systems include antioxidant enzymes, free radical scavengers, and metal chelators. The purpose of this review is to examine the potential of nonessential dietary components that inhibit oxidative reactions in foods and biological tissues.

Phenolics: prooxidants or antioxidants?

Both essential and nonessential dietary antioxidants have been suggested to be beneficial for health. However, studies show that many antioxidants can also exhibit prooxidant behavior under certain conditions. The antioxidant/prooxidant activity of phenolics is dependent on such factors as metal-reducing potential, chelating behavior, pH, and solubility characteristics. These factors, in addition to bioavailability and stability in tissues, should be considered when evaluating the potential bioactivity of dietary phenolics.

The role of stereospecific saturated fatty acid positions on lipid nutrition.

The saturated fatty acids in the sn-1 and -3 position of triacylglycerols can exhibit different metabolic patterns due to their low absorptivity. This means that dietary fats containing saturated fats primarily in sn-1 and -3 positions (e.g., cocoa butter, coconut oil, and palm oil) can have very different biological consequences than those fats in which the saturated fats are primarily in the sn-2 position (e.g., milk fat and lard). Differences in stereospecific fatty acid location should therefore be an important consideration in the design and interpretation of lipid nutrition studies and in the production of specialty food products.

High-fat meals and endothelial function.

Two recent studies on the effects of single high-fat meals on endothelial function have postulated an additional mechanism by which dietary lipids can impact atherosclerosis. These studies found that dietary lipids decrease the ability of blood vessels to dilate and that this modification of endothelial function was protected by dietary antioxidants. Although this research presents an interesting hypothesis, much more research is necessary to determine the importance of dietary factors on vasoactivity.

Ability of carnosine and other skeletal muscle components to quench unsaturated aldehydic lipid oxidation products.

Breakdown of lipid peroxides results in the formation of aldehydic compounds which are toxic to biological systems and deleterious to food quality. To determine the potential of skeletal muscle compounds to protect biomolecules from lipid oxidation products, the ability of carnosine and various other related compounds to quench monounsaturated and polyunsaturated aldehydes was investigated. Carnosine, the most abundant dipeptide in skeletal muscle, is capable of quenching alpha,beta-monounsaturated aldehydes and 4-hydroxy-2-trans-nonenal (HNE) more effectively than its constituent amino acid. Carnosine (5 mM) reduced 44% of headspace trans-2-hexenal (0.5 mM) after 1 h incubation at 40 degrees C and pH 7.4. Other histidine-containing dipeptides and the amine compounds, spermine and spermidine, had similar or slightly lower quenching activity than carnosine. Glutathione and thioctic acid had superior quenching ability than carnosine, but their overall contribution to aldehyde quenching compared to carnosine is limited due to their lower concentration in skeletal muscle. The results suggest that carnosine could be important for decreasing the toxicity of lipid oxidation products in biological systems and for minimizing rancidity in muscle foods.

Ability of amino acids, dipeptides, polyamines, and sulfhydryls to quench hexanal, a saturated aldehydic lipid oxidation product.

Hexanal is a common product arising from the oxidation of omega-6 fatty acids. Because aldehydic lipid oxidation products can react with food components, interactions between hexanal and sulfhydryl- and amine-containing compounds were determined. The polyamines, spermidine and spermine, and the sulfhydryl-containing compounds, glutathione and thioctic acid, decreased headspace hexanal concentrations < or =7.0%. Histidine was the only amino acid tested that was able to quench headspace hexanal. Histidine-containing dipeptides decreased headspace hexanal 3.0-8.5-fold more than histidine. Hexanal quenching by the hisitidine-containing dipeptides increased as the size of the aliphatic side group of the amino acid adjacent to histidine increased, with Leu-His having the greatest hexanal quenching activity. The ability of Leu-His to quench histidine increased with increasing pH. The ability of histidine-containing dipeptides to interact with hexanal suggests that it may be possible to design peptides to alter the concentration of saturated aldehydes in oxidizing lipids.

Peroxynitrite-induced oxidation of lipids: implications for muscle foods.

Peroxynitrite (ONOO(-)), formed from the nearly diffusion limited reaction between nitric oxide and superoxide, could be an important prooxidant in muscle foods. The objective of this study was to determine whether peroxynitrite caused oxidation of pyrogallol red, liposomes, muscle microsomes, and skeletal muscle homogenate. Oxidation of pyrogallol red, liposomes, and microsomes initiated by peroxynitrite continuously produced by 3-morpholinosydnonimine (SIN-1, 2 mM) was time-dependent and enhanced by CO(2) (1 mM). Reagent peroxynitrite (2 mM) caused concentration-dependent oxidation of pyrogallol red, liposomes, and muscle microsomes that was very rapid with no change after 5 min. Peroxynitrite-induced oxidation was suppressed by CO(2) and low pH. Skeletal muscle homogenate oxidized by reagent peroxynitrite (0.5 mM) exhibited gradual oxidation with time and was suppressed by CO(2), low pH, and metal chelators. These data suggest that peroxynitrite could be an important prooxidant in muscle foods.

Lipid oxidation in emulsions as affected by charge status of antioxidants and emulsion droplets.

The influence of charge status of both lipid emulsion droplets and phenolic antioxidants on lipid oxidation rates was evaluated using anionic sodium dodecyl sulfate (SDS) and nonionic polyoxyethylene 10 lauryl ether (Brij)-stabilized emulsion droplets and the structurally similar phenolic antioxidants gallamide, methyl gallate, and gallic acid. In nonionic, Brij-stabilized salmon oil emulsions at pH 7.0, gallyol derivatives (5 and 500 microM) inhibited lipid oxidation with methyl gallate > gallamide > gallic acid. In the Brij-stabilized salmon oil emulsions at pH 3.0, low concentrations of the galloyl derivatives were prooxidative or ineffective while high concentrations were antioxidative. In SDS-stabilized salmon oil emulsions, oxidation rates were faster and the galloyl derivatives were less effective compared to the Brij-stabilized emulsions. Differences in antioxidant activity were related to differences in the ability of the galloyl derivatives to partition into emulsion droplets and to increase the prooxidant activity of iron at low pH.

Impact of dietary conjugated linoleic acid on the oxidative stability of rat liver microsomes and skeletal muscle homogenates.

Dietary conjugated linoleic acid (CLA; 0-2.0%) increased CLA concentrations in liver microsomes and skeletal muscle homogenates from rats. Dietary CLA decreased oleic and arachadonic acid concentrations in both liver microsomes and skeletal muscle. The presence of CLA in liver microsomes had no impact on linoleic acid, arachadonic acid, and alpha-tocopherol oxidation rates. Dietary CLA (2.0%) also did not alter alpha-tocopherol oxidation rates in liver microsomes or muscle homogenates. Formation of malonaldehyde (MDA) in oxidizing liver microsomes decreased with increasing CLA concentration as determined by measurement of thiobarbituric acid-MDA complexes by HPLC. The ability of CLA to decrease MDA formation without impacting other lipid oxidation markers such as the disappearance of fatty acid and alpha-tocopherol suggests that decreased MDA concentration was the result of CLA's ability to lower polyenoic fatty acids such as arachadonic acid. While CLA does not appear to act as an antioxidant, its ability to decrease polyenoic fatty acid concentrations could decrease the formation of highly cytotoxic lipid oxidation products such as MDA.

Ability of surfactant hydrophobic tail group size to alter lipid oxidation in oil-in-water emulsions.

Oxidation of oil-in-water emulsion droplets is influenced by the properties of the interfacial membrane surrounding the lipid core. Previous work has shown that an important factor in the oxidation of oil-in-water emulsions is surfactant properties that impact interactions between water-soluble prooxidants and lipids in the emulsion droplet. The purpose of this research was to study the impact of surfactant hydrophobic tail group size on lipid oxidation in oil-in-water emulsions stabilized by polyoxyethylene 10 lauryl ether (Brij-lauryl) or polyoxyethylene 10 stearyl ether (Brij-stearyl). The ability of iron to decompose cumene peroxide was similar in hexadecane emulsions stabilized by Brij-stearyl and Brij-lauryl. Oxidation of methyl linoleate in hexadecane emulsions containing cumene peroxide was greater in droplets stabilized by Brij-lauryl than in those stabilized by Brij-stearyl at pH 3 with no differences observed at pH 7.0. Oxidation of salmon oil was greater in emulsions stabilized by Brij-lauryl than in those stabilized by Brij-stearyl as determined by both lipid peroxides and headspace propanal. These results suggest that surfactant hydrophobic tail group size may play a minor role in lipid oxidation in oil-in-water emulsions.

Mechanisms of the antioxidant activity of a high molecular weight fraction of whey.

The antioxidant mechanisms of whey proteins in a Tween 20-stabilized salmon oil-in-water emulsion were investigated. The antioxidant activity of the high molecular weight (HMW) fraction of whey from pasteurized milk was found to increase with concentration, as determined by its ability to inhibit TBARS and lipid peroxide formation. The ability of sulfhydryl-blocked whey to inhibit TBARS formation was reduced 60% compared to the HMW fraction alone at 7 days of storage. HMW fraction was able to scavenge peroxyl radicals, with scavenging decreasing approximately 20% when sulfhydryls were blocked. HMW fraction was able to chelate iron away from the surface of negatively charged BSA-stabilized emulsion droplets, indicating that the whey proteins were able to chelate iron. A better understanding of the mechanisms by which whey proteins inhibit lipid oxidation could increase the use of whey proteins as food antioxidants.

Iron-accelerated cumene hydroperoxide decomposition in hexadecane and trilaurin emulsions.

Free radicals arising from lipid peroxides accelerate the oxidative deterioration of foods. To elucidate how lipid peroxides impact oxidative reactions in food emulsions, the stability of cumene hydroperoxide was studied in hexadecane or trilaurin emulsions stabilized by anionic (sodium dodecyl sulfate; SDS), nonionic (Tween 20), and cationic (dodecyltrimethylammonium bromide; DTAB) surfactants. Fe(2+) rapidly (within 10 min) decomposed between 10 and 31% of the cumene hydroperoxide in Tween 20- and DTAB-stabilized emulsions at pH 3.0 and 7.0 and in the SDS-stabilized emulsion at pH 7.0 with no further decomposition of peroxides occurring for up to 3 h. In SDS-stabilized emulsions at pH 3.0, Fe(2+) decreased peroxides by 90% after 3 h. Decomposition of peroxides in the absence of added iron and by Fe(3+) was observed only in SDS-stabilized emulsions at pH 3.0. These results suggest that peroxide decomposition by iron redox cycling occurs when iron emulsion droplet interactions are high.

The effects of surfactant type, pH, and chelators on the oxidation of salmon oil-in-water emulsions.

Lipid oxidation in emulsions is influenced by the ability of transition metals to associate with emulsion droplets. The oxidative stability of 5% salmon oil-in-water emulsion was influenced by surfactant type, with oxidation rates being greatest in emulsions stabilized by anionic sodium dodecyl sulfate (SDS) followed by nonionic Tween 20 and cationic dodecyltrimethylammonium bromide (DTAB). EDTA inhibited lipid oxidation in all the emulsions, and apo-transferrin inhibited oxidation in the Tween 20-stabilized emulsions at pH 7.0, suggesting that continuous-phase iron was an active prooxidant. Iron associated with Tween-20 stabilized hexadecane emulsion droplets could be partitioned into the continuous phase by lowering the pH to

Ability of iron to promote surfactant peroxide decomposition and oxidize alpha-tocopherol.

Peroxides are an important factor in oxidative reactions in foods because their decomposition can result in formation of highly reactive free radicals. Emulsifiers such as the Brijs, Tweens, and lecithin were found to contain 4-35 micromol of peroxides/g of surfactant. Peroxide concentrations in Tween 20 micelles increased in the presence of low iron concentrations but decreased when iron concentrations were high, suggesting that iron was capable of promoting both peroxide formation and decomposition. Oxidation of alpha-tocopherol was observed in micelles high in peroxides (Tween 20) but not in micelles where peroxide concentrations were low (Brij). Transition metals accelerated the oxidation of alpha-tocopherol in Tween 20 micelles, whereas EDTA stabilized alpha-tocopherol in the presence of added Fe(2+). These results suggest that surfactant peroxides could decrease the oxidative stability of food emulsions by acting as a source of free radicals, especially in the presence of transition metals.

Inhibition of low-density lipoprotein oxidation by carnosine histidine.

Carnosine is a beta-alanylhistidine dipeptide found in skeletal muscle and nervous tissue that has been reported to possess antioxidant activity. Carnosine is a potential dietary antioxidant because it is absorbed into plasma intact. This research investigated the ability of carnosine to inhibit the oxidation of low-density lipoprotein (LDL) in comparison to its constituent amino acid, histidine. Carnosine (3 microM) inhibited Cu2+-promoted LDL (20 of protein/mL) oxidation at carnosine/copper ratios as low as 1:1, as determined by loss of tryptophan fluorescence and formation of conjugated dienes. Carnosine (6 microM) lost its ability to inhibit conjugated diene formation and tryptophan oxidation after 2 and 4 h of incubation, respectively, of LDL with 3 microM Cu2+. Compared to controls, histidine (3 microM) inhibited tryptophan oxidation and conjugated diene formation 36 and 58%, respectively, compared to 21 and 0% for carnosine (3 microM) after 3 h of oxidation. Histidine was more effective at inhibiting copper-promoted formation of carbonyls on bovine serum albumin than carnosine, but carnosine was more effective at inhibiting copper-induced ascorbic acid oxidation than histidine. Neither carnosine nor histidine was a strong inhibitor of 2,2'-azobis(2-amidinopropane) dihydrochloride-promoted oxidation of LDL, indicating that their main antioxidant mechanism is through copper chelation.

Impact of tween 20 hydroperoxides and iron on the oxidation of methyl linoleate and salmon oil dispersions.

To determine the role of surfactant hydroperoxides on the oxidative stability of fatty acids, the oxidation of methyl linoleate micelles and salmon oil-in-water emulsions was measured as a function of varying Tween 20 hydroperoxide concentrations. Increasing Tween 20 hydroperoxide concentrations from 3.5 to 14.7 micromol hydroperoxide/g Tween 20 decreased the lag phase of headspace hexanal formation but did not increase the total amount of hexanal formed in methyl linoleate/Tween 20 micelles. In the micelle system, Fe(2+) decreased the lag phase of hexanal formation but increased total hexanal concentrations only in micelles with the highest Tween 20 hydroperoxide concentrations (14.7 micromol hydroperoxide/g surfactant). Increasing Tween 20 surfactant hydroperoxide concentrations also increased the oxidation of salmon oil-in-water emulsions as determined by lipid hydroperoxides and headspace propanal. In both the micelle and emulsion systems, the prooxidant effect of Fe(2+) decreased with increasing Tween 20 hydroperoxide concentrations. These data show that surfactant hydroperoxides such as those in Tween 20 could decrease the oxidative stability of lipids in food emulsions.