Effect of ascorbyl palmitate on the preservation of α-tocopherol in sunflower oil,alone and with herbs and spices

Ascorbyl palmitate (AP) (200 ppm) preserved α-tocopherol in sunflower oil at 95°C and delayed the onset of rancidity. Both effects increased with AP concentration (r² = 0.96 and 0.97 respectively) but levelled off near ∼700 ppm. The improved anti-rancidity effect was due to the increased preservation (P<0.01). Synergism was observed for both effects for AP combined with sage, turmeric, oregano and marjoram. Clove and thyme gave a smaller synergistic effect whereas basil inhibited. Neither bay nor cumin had any effect. Both the preservative (PF_p) and anti-rancidity effects (PF_r) were directly related to the thyme concentration (0–2000 ppm). Again, the decreased rancidity was due to the increased preservation (P<0.01). The optimum AP concentration (0–1000 ppm) was around 250 ppm (P<0.01) with thyme present (at 500 ppm) (r²=0.99). The increased delay in rancidity was due to the improved preservation of α-tocopherol (r=1, r² = 0.99, P<0.01). Both the logarithm of the induction time and the preservative effect for the mixture of thyme and AP was directly related to the temperature (80–105°C). The mode of action of AP is also discussed.     

Effects of chitosan and collagen containing α-tocopherol on the oxidative stability in bulk oil and oil-in-water emulsion

To provide efficient antioxidant capacities, proper carriers are needed to protect antioxidants against oxidative stress. Collagen mesh structure or chitosan gel was loaded with α-tocopherol and their effects were evaluated in bulk corn oil or oil-in-water (O/W) emulsion at 60 °C. Added collagen and chitosan enhanced oxidative stability in corn oil and O/W emulsions at 60 °C compared to corn oils without carriers or with addition of α-tocopherol (p < 0.05). Stability of α-tocopherol in corn oil loaded in collagen or chitosan was significantly enhanced compared to that in oils without carriers (p < 0.05). In O/W emulsions, α-tocopherol loaded collagen showed higher antioxidant properties than α-tocopherol loaded chitosan (p < 0.05). Collagen mesh structure and chitosan gel retarded the rates of lipid oxidation efficiently in both food matrices when α-tocopherol was not loaded. Collagen mesh structure and chitosan gel can be useful carriers for α-tocopherol in bulk oil or O/W emulsion.     

Effects of rendering and α-tocopherol addition on the oxidative stability of horse fat

Food Science and Biotechnology - Oxidative stability of horse fat rendered at 70&nbsp;°C, − 0.1&nbsp;MPa under vacuum and 110&nbsp;°C, 0.1&nbsp;MPa from horse...     

Effects of catechin and α-tocopherol addition on the autoxidative stability of diacylglycerol oil derived from an olive oil and perilla oil mixture

The effects of tea catechin and α-tocopherol addition (1 mM) on the oxidative stability of oleic and linolenic acid-rich diacylglycerol (DAG) oil derived from an extra virgin olive oil and perilla oil mixture (6:4, w/w) were evaluated. Oil was oxidized at 50°C for 10 days, after which oxidation was evaluated based on headspace oxygen consumption and peroxide values (POV). The polyphenol and tocopherol contents were also monitored. Addition of catechin did not affect the oxygen consumption or POV of DAG oil, whereas α-tocopherol acted as a prooxidant. Addition of antioxidants had no significant (p>0.05) effect on the fatty acid composition of the oil. Degradation of γ-tocopherol during oil oxidation was inhibited by addition of α-tocopherol, and addition of antioxidants inhibited polyphenol degradation. The autoxidative stability of DAG oil can be improved using polar rather than non-polar antioxidants.     

Antioxidant capacities of α-tocopherol,trolox, ascorbic acid,and ascorbyl palmitate in riboflavin photosensitized oil-in-water emulsions

Antioxidant capacities of α-tocopherol, trolox, ascorbic acid, and ascorbyl palmitate at 0.01, 0.1, and 1.0 mM in riboflavin photosensitized oil-in-water (O/W) emulsions were determined using headspace oxygen depletion, lipid hydroperoxide, and headspace volatile analyses. After 32 h visible light irradiation, headspace oxygen in O/W emulsions without adding antioxidants, with 1.0 mM α-tocopherol, trolox, ascorbic acid, and ascorbyl palmitate decreased to 18.50%, 18.85%, 16.01%, 17.92%, and 19.88%, respectively, whereas those samples in the dark were 20.74%. Trolox and ascorbic acid acted as prooxidants while their lipophilic counterparts, α-tocopherol and ascorbyl palmitate, respectively showed antioxidant properties. Similar antioxidative or prooxidative properties of the tested compounds can be observed in the results of lipid hydroperoxides and headspace volatiles. However, the prooxidant and antioxidant properties of the tested compounds were not clearly shown at 0.01 and 0.1 mM concentration. Both the type and concentration of antioxidants influenced the antioxidant capacities in riboflavin photosensitized O/W emulsions.     

Effect of tocopherols,tocotrienols, β-carotene,and chlorophyll on the photo-oxidative stability of red palm oil

Effect of tocols, β-carotene, and chlorophyll on photo-oxidative stability of red palm oil (RPO) were studied. Model systems of triacylglycerols+tocols, triacylglycerols+β-carotene, triacylglycerols +tocols+β-carotene, and triacylglycerols+tocols+β-carotene+chlorophyll were exposed to fluorescent light at intensities of 5,000, 10,000, and 15,000 lux for 7 h at 30±2°C. Changes in concentrations of tocopherols, tocotrienols, β-carotene, chlorophyll, and peroxide values were evaluated every hour. Light intensity accelerated degradation of tocols in the triacylglycerols+tocols system and β-carotene in the triacylglycerols+β-carotene system. Gamma-tocotrienol showed the highest degradation rate and β-carotene was the most sensitive compound to changes in light intensity, indicated by the lowest light intensity coefficient (z_i) value. The presence of tocols and β-carotene together showed protective effects for the photo-oxidative stability of RPO. The presence of chlorophyll increased the rate of photo-oxidation at high light intensities. Interactions between tocols and β-carotene contributed to the photo-oxidative stability of RPO.     

Effects of α-tocopherol on the oxidative stability and incorporation of deuterium in volatiles from a linoleic acid-deuterium model system

The effects of α-tocopherol on the oxidative stability and incorporation of deuterium in volatiles were evaluated in linoleic acid-water model systems treated at 60°C by analyzing headspace oxygen depletion, formation of lipid hydroperoxides, and profiles of headspace volatiles. Deuterium oxide accelerated the rates of linoleic acid oxidation compared to samples in deuterium-free water. As the concentration of α-tocopherol increased from 0 to 1500 ppm, the consumption of headspace oxygen and the formation of volatiles decreased, whereas the contents of lipid hydroperoxides did not decrease in the linoleic acid-water system. The mass to charge ratios (m/z) of volatiles in linoleic aciddeuterium oxide were significantly higher than those with deuterium oxide-free water. Generally, the presence of α-tocopherol decreased the mass to charge ratios (m/z) of volatiles including pentanal, hexanal, t-2-heptenal, and 2-octenal, implying that α-tocopherol may be involved in the aldehyde formation from lipid oxidation.     

Effects of α-tocopherol and citric acid on the oxidative stability of alimentary poultry fats during storage at low temperatures

The purpose of this study was to investigate the stability of three alimentary poultry fats (goose, duck, and chicken) by natural antioxidants (α-tocopherol and citric acid). This was targeted to extend their shelf life, and to monitor the quality parameters during refrigerated (+4°C) and frozen storage (–20°C). The addition of natural antioxidants in a proportion of 0.1% has extended the shelf life of goose fat stored at +4°C by 90 days; for goose fat stored at –20°C citric acid has prolonged the shelf life by 150 days, while goose fat with α-tocopherol could be stored for more than 480?days at –20°C without spoilage. Polyunsaturated fatty acids and monounsaturated fatty acids content decreased significantly (p < 0.05) after 480 days of chilled storage for fat samples with α-tocopherol. The natural antioxidants provided good protection against oxidation of poultry fats, and these can be used to monitor the oxidation of fats and to predict their shelf life stability.     

Influence of α-, γ-, and δ-tocopherol on the radiation induced formation of peroxides in rapeseed oil triacylglycerols

The effect of α-, γ-, and δ-tocopherols (enrichment: 1000 ppm) on the peroxide formation in rapeseed oil triacylglycerols (RSOTG) was evaluated. The oxidation process was initiated by gamma-irradiation with doses of 4 and 10 kGy. Whereas a pronounced antioxidant effect was observed for γ- and δ-tocopherol (sequence: δ- > γ-tocopherol), the inhibition extent of α-tocopherol was insignificant.     

Oxidative stability of oil-in-water emulsions with α-tocopherol,charged emulsifier,and different oxidative stress

α-Tocopherol is known to show different activity depending on the concentration and food matrix. Effects of α-tocopherol at the concentrations of 0, 0.1, 0.5, and 1.0 mM were determined in oil-in-water (O/W) emulsions containing anionic, neutral, and cationic emulsifiers under different types of oxidative stress including riboflavin photosensitization, photooxidation, and autoxidation. Headspace oxygen depletion, lipid hydroperoxides, and conjugated dienes were analyzed to determine the oxidative stability of O/W emulsions. α-Tocopherol served as an antioxidant in O/W emulsion with a cationic emulsifier irrespective of oxidative stress. α-Tocopherol acted as an antioxidant in O/W emulsion with a neutral emulsifier at riboflavin photosensitization while a prooxidant at photooxidation. However, in samples with an anionic emulsifier, α-tocopherol activity differed from the concentration and types of oxidative stress. Therefore, cationic transition metals or reactive oxygen species generated from RF photosensitization could play key roles of α-tocopherol in O/W emulsion.     

Effects of dietary α-tocopheryl acetate supplementation on α-tocopherol deposition in porcine m. psoas major and m. longissimus dorsi and on drip loss, colour stability and oxidative stability of pork meat

The effect of feeding supra-nutritional levels of α-tocopheryl acetate on its deposition in two porcine muscles of different metabolic rates (m. longissimus dorsi and m. psoas major) and the effect on meat quality (lipid oxidation, colour stability and drip loss) was studied. Pigs were fed a standard diet supplemented with three levels: 100, 200 and 700 mg/kg of α-tocopheryl acetate from the time of weaning to slaughter at 90kg live weight. Muscle α-tocopherol levels were linearly related to the logarithm of dietary α-tocopheryl acetate supplementation and the linear relationship was estimated for the two muscles. The levels of α-tocopherol in the two muscles differed by a parallel displacement with consistently higher α-tocopherol levels in m. psoas major compared to m. longissimus dorsi. Dietary α-tocopheryl acetate supplementation significantly reduced lipid oxidation as measured by thiobarbituric acid reactive substances (TBARS) in both raw and cooked meat during storage at 4 °C for 6 days. Drip loss and colour stability of raw muscles were not affected by dietary α-tocopheryl acetate levels, 100mg α-tocopheryl acetate/ kg feed resulted in sufficient α-tocopherol levels in muscles to ensure minimum drip loss and optimum colour stability.     

Effect of Sicilian pasture feeding management on content of α-tocopherol and β-carotene in cow milk

This study was performed to evaluate α-tocopherol and β-carotene contents of pasture milk under ordinary Sicilian farming conditions. Fourteen dairy farms were allocated into 2 balanced groups on the basis of cultivated (CULT) or spontaneous (SPO) pasture type feeding. Bulk milk per farm was collected 4 times from February through April at 3-wk intervals. Pasture botanical and diet composition, diet nutritional quality, milk yield and composition were estimated each time. Pasture intake levels were calculated based on feed analyses, hay and concentrate amounts fed, and milk yield and chemical composition. According to pasture intake, the farms were split into low pasture intake (LPI; <29.5% of dry matter) and high pasture intake (HPI; >29.5% of dry matter) groups. Milk samples per farm were analyzed for α-tocopherol and β-carotene contents by HPLC. The SPO group had higher levels of α-tocopherol and β-carotene in milk (0.7 and 0.3 mg/L, respectively) and milk fat (19.0 and 7.5 mg/kg fat, respectively) compared with the CULT group in milk (0.5 and 0.2 mg/L, respectively) and milk fat (14.6 and 4.9 mg/kg, respectively). High pasture intake compared with LPI increased α-tocopherol in milk fat (18.0 and 16.0 mg/kg of fat, respectively). However, only in the SPO (not in CULT), HPI compared with LPI increased milk α-tocopherol (0.8 vs. 0.6 mg/L, respectively), milk β-carotene (0.3 vs. 0.2 mg/L, respectively), and milk fat β-carotene (8.4 vs. 6.6 mg/kg, respectively). Results may be related to the different botanical composition of the respective pasture types and pasture intake. Spontaneous pasture compared with CULT contained a higher mass proportion of Asteraceae, Fabaceae, Cruciferae, Euphorbiaceae, and Malvaceae plants. Milk and milk fat α-tocopherol levels were higher on test-days (TD)-1, TD-2, and TD-4 compared with TD-3. For HPI farms, milk fat β-carotene content was higher on the first 2 TD compared with the last 2 TD. These differences could be related to plant biological stage. On Sicilian dairy farms, the highest milk α-tocopherol and β-carotene contents may be obtained feeding high levels of SPO pasture in the spring.     

Effects of prepartum supplementation of β-carotene on colostrum and calves

Little is known about transfer of dietary β-carotene into colostrum, its absorption by the calf, and its effects on retinol and α-tocopherol in the calf when the dam's dietary vitamin A is adequate. Our objective was to assess the effect of β-carotene supplementation during the close-up dry period on the colostrum and calf. The study was conducted on a large commercial dairy farm in Indiana during early summer of 2015. Ninety-four multiparous Holstein cows were blocked by calving data, parity, and previous production, and then randomly assigned to either control or β-carotene (BC) treatments. While locked in headgates each morning, each cow received a topdress of β-carotene (Rovimix, DSM Nutritional Products, 8 g/d; provided 800 mg β-carotene) or carrier from 21 d before expected calving until calving. Colostrum was collected within 2 h of parturition. Calf blood samples were obtained within 2 h of birth before receiving the dam's colostrum, at 24 h after birth, and at 7 d and 60 d of age. Blood serum was analyzed for β-carotene, retinol, α-tocopherol, and other metabolites and enzymes. Colostrum was analyzed for β-carotene, retinol, α-tocopherol, colorimetry profile, and milk components. Data were analyzed using mixed-effects models in SAS (SAS Institute Inc.). Calf serum β-carotene data were analyzed using the FREQ procedure. Colostrum β-carotene was higher for BC cows. Colostrum from BC cows had increased a* [measures red (positive) to green (negative)] and b* [measures yellow (positive) to blue (negative)] colorimeter values, indicating that β-carotene altered colostrum color toward red and yellow. Supplementation did not affect colostral or calf IgG concentrations. Colostrum color indices were correlated with IgG concentrations as well as concentrations of β-carotene, retinol, and α-tocopherol. Before receiving colostrum, the concentration of β-carotene in calf serum was below the detectable threshold of 0.05 μg/mL. At 24 h of age, the number of calves with detectable β-carotene concentrations increased, with more calves from BC cows (52.1%) having detectable concentrations than calves from cows in the control group (6.1%). No differences in concentrations of retinol or α-tocopherol were observed in calf serum. Supplementation of β-carotene to cows decreased activities of gamma-glutamyl transpeptidase and glutamate dehydrogenase in calf serum. In pregnant cows already receiving adequate vitamin A, supplementation of β-carotene increased concentration of β-carotene in colostrum, altered colostrum color, and increased serum β-carotene in calves at birth.     

Effect of α- and δ-tocopherol on the oxidative stability of a mixed hydrogenated fat under frying conditions

Information on the antioxidative potential of -tocopherol is scanty, in particular for frying conditions. This study was aimed at assessing the antioxidative effects and degradation of - and -tocopherol between 0.01 and 0.1% on the oxidation of a commercial frying fat at 160 °C. Oxidation experiments were performed by assessing every 15 or 30 min the peroxide value and conjugated dienes as primary oxidation products, p-anisidine reactive products and hexanal as secondary oxidation products, as well as the stability of tocopherols. The fatty acid composition was determined after 6 h. The fat samples enriched with -tocopherol were considerably less resistant to oxidation compared to those with -tocopherol. Both primary and secondary oxidation parameters increased their speed of formation with -tocopherol but not with -tocopherol. The latter observation is confirmed for the total amount of oxidation and also for the first stage, showing a lag phase only for -tocopherol. These antioxidative effects can be due to the higher stability of -tocopherol compared to -tocopherol, which is oxidized faster into tocopheryl radicals, which can participate in side reactions that result in an acceleration of the oxidation speed. Neither -tocopherol nor -tocopherol influenced the fatty acid pattern. The investigation showed that during mild frying conditions the tocopherol homologues displayed various antioxidant activities. The less effective homologue, -tocopherol, underwent disintegration more quickly than -tocopherol.     

Effect of vitamin E supplementation on α-tocopherol and β-carotene concentrations in tissues from pasture- and grain-fed cattle

The effects of dietary vitamin E supplementation of grain-fed cattle on lipid oxidation and meat colour have been extensively investigated, but little attention has been given to pasture-fed cattle where meat is likely to contain naturally high amounts of α-tocopherol and carotenoids. In the work described, we evaluated the effects of pasture-feeding alone and with vitamin E supplementation on tissue levels of anti-oxidants and compared the findings with those obtained for grain-fed cattle with and without supplementation. Sorghum was the major component of the grained-based ration. α-Tocopherol concentrations in plasma, muscle and fat tissues of pasture-fed cattle were not affected by vitamin E supplementation (2500 IU/head/day for 132 days prior to slaughter) while those of grain-fed cattle increased significantly. The α-tocopherol concentrations in the supplemented grain-fed cattle were similar in muscle and liver to pasture-fed animals but were lower in their fat (P<0.05). The major carotenoid present in all tissues studied from pasture-fed was β-carotene and its contents in plasma, liver, fat and muscles were decreased (P<0.05) by supplementation with vitamin E. Carotenoids were essentially absent in grain-fed cattle except for small amounts in liver. The implication of this study for the meat industry is that cattle grazed on good pasture can achieve concentrations of α-tocopherol in muscles and other tissues at least as high as those obtained by supra-nutritional supplementation of grain-fed cattle with vitamin E. However, α-tocopherol supplementation of pasture-fed cattle reduced tissue concentrations of β-carotene, which would reduce carcase fat yellowness and make pasture-fed cattle more acceptable to some Asian markets.     

The effect of oxygen level and exogenous α-tocopherol on the oxidative stability of minced beef in modified atmosphere packs

The effect of oxygen level (20, 40, 60 and 80%) in modified atmospheres on the oxymyoglobin content of intact and minced beef (M. semimembranosus, SM) was evaluated. There was no significant difference in the oxymyoglobin content of minced SM stored for up to 4 days in modified atmosphere packs containing 20, 40, 60 or 80% O(2). After 7 days, oxymyoglobin in minced SM decreased significantly (P?0.05) with decreasing oxygen level but by day 10 all samples had similarly low oxymyoglobin contents. Lipid oxidation increased significantly (P?0.05) between day 7 and 10 of storage in minced SM stored in modified atmospheres containing 40, 60 or 80% O(2). Oxymyoglobin and lipid oxidation occurred in intact SM but the extent of oxidation was lower than for minced SM. Exogenous α-tocopherol, dispersed in olive oil and added to minced SM (300 and 3000 mg α-tocopherol/kg lipid), had no significant effect on Hunter 'a' values when the samples were stored in low (20%) or high (80%) oxygen atmospheres. Exogenous α-tocopherol addition led to a significant reduction in lipid oxidation (P?0.05) in minced SM stored in high but not in low oxygen atmospheres.     

Influence of dietary β-alanine and histidine on the oxidative stability of pork

Carnosine (β-alanine-L-histidine) and anserine (β-alanine-1-methyl histidine) are endogenous antioxidants found in skeletal muscle. The objective of this research was to determine if supplementation of swine diets with histidine (histidine; 0.40%) and/or β-alanine (β-alanine; 0.225%) was an effective method to increase carnosine and anserine concentrations and the oxidative stability of Longissimus dorsi (LD) and Vastus intermedius (VI) muscles. Dietary treatments had no effect on carnosine and anserine concentrations in LD; however, histidine + β-alanine supplementation increased carnosine and anserine in VI muscle compared to β-alanine supplementation. Dietary supplementation had no effect on the formation of thiobarbituric acid reactive (TBARS) or lipid peroxides in cooked VI and LD. In salted VI and LD muscle, differences in TBARS and peroxides were observed; however, these differences did not consistently correlate with differences in anserine and carnosine concentrations. Therefore, the results of this research suggest that supplementation of swine diets with β-alanine and/or histidine is not an efficient method to increase the oxidative stability of pork.     

Effect of dietary α-tocopherol supplementation on color and lipid stability in pork

Myoglobin and lipid oxidation are major causes of quality deterioration in fresh pork. A process to enhance color and lipid stability would prove valuable to the pork industry given the current trend of centralized packaging and distribution to retail markets. Our objective was to determine the effects of dietary α-tocopherol (α-Toc) supplementation on color and lipid stability in ground pork, and loin chops stored in modified atmosphere packaging (MAP). Yorkshire crossbred pigs (n=20) were randomized into two groups and fed diets containing 48 (CON) or 170 mg α-Toc acetate/kg feed (VIT-E) for 6 weeks before slaughter. Plasma α-Toc concentration was measured weekly. Post-slaughter, Boston butt shoulders were ground, formed into patties with or without 1.5% salt, and stored fresh at 4°C for 0, 2, 4, or 6 days, and frozen at −20°C for 45 or 90 days. Pork loin chops were packaged aerobically and stored at 4°C for 0, 2, 4 or 6 days, or in MAP at 4°C for 7, 10 or 13 days prior to Hunter L*,a*,b* and TBARS analyses. α-Toc concentration of longissimus dorsi, psoas major, biceps femoris, semimembranosus and semitendinosus muscles was determined. Plasma α-Toc was greater (P<0.05) in VIT-E animals compared with CON and α-Toc concentrations were greater (P<0.05) in all VIT-E muscles compared with CON. TBARS values of both fresh and salted patties were less in VIT-E than in CON meat following 6 days at 4°C; VIT-E TBARS of salted patties were less (P<0.05) after 45 days at −20°C compared with CON. α-Toc supplementation did not influence (P>0.05) color of aerobically packaged or MAP chops, or of fresh or salted pork patties. α-Toc supplementation reduced TBARS formation in fresh and salted pork but had no significant impact on color.     

Supranutritional vitamin E supplementation in pigs: Influence on subcellular deposition of α-tocopherol and on oxidative stability by conventional and derivative spectrophotometry

The influence of three levels of vitamin E (30, 200 and 1000mg kg⁻¹) in the diet of pigs on the subcellular deposition of α-tocopherol in muscle and on the oxidative stability determined by conventional and first derivative Spectrophotometry was studied. The content of α-tocopherol in m. gluteo biceps and in mitochondrial and microsomal fractions of the muscle significantly increased (p < 0.01) with increasing levels of dietary vitamin E. Concentrations of α-tocopherol in muscle, mitochondria and microsomes of pigs fed diet supplemented with 1000 mg kg⁻¹ α-tocopheryl acetate were 3.2-, 6.1- and 5.6-fold greater, respectively, than those in their counterparts from the control animals. These differences in α-tocopherol concentration in the subcellular fractions and intact muscle resulted in enhanced stability of the membranes and the tissue when exposed to iron-ascorbate induced peroxidation. When lipid oxidation in the same samples was further measured by the first derivative method, the resultant MDA-TBA values were 59–69% lower in tissue samples, 16–19% lower in mitochondria and 6–9% lower in microsomes than the conventional TBARS values.     

Effect of dietary rosemary and α-tocopheryl acetate on the oxidative stability of raw and cooked pork following oxidized linseed oil administration

The effect of a 2% dietary administration to pigs of oxidized linseed oil (targeted level of 150mEq.O(2)/kg oil after heating at 50°C and exposure to air for 3-4 days following addition of 10ppm CuSO(4)), either or not in combination with antioxidants, on the oxidative stability of raw and cooked pork during illuminated chill storage was assessed. The antioxidant treatments were: 40ppm α-tocopheryl acetate, 40ppm rosemary extract, 40ppm rosemary extract+2ppm gallic acid, and 40ppm α-tocopheryl acetate+40ppm rosemary extract. A total of 20ppm of α-tocopheryl acetate (ATA) was added to all diets in order to meet the physiological requirement of the animals. The antioxidant treatments did not exert any effect on colour and protein oxidation. Lipid oxidation was only decreased by dietary ATA when comparing the ATA supplemented groups combined versus a control treatment group for raw but not for cooked meat. This was due to a higher content of α-tocopherol in the meat and subcutaneous fat. The lipid oxidation results suggested a lack of antioxidant effect for the rosemary extract. No evidence for a synergistic effect of the antioxidant combinations was observed.