Impact of thermal processing on the antioxidant mechanisms of continuous phase β-lactoglobulin in oil-in-water emulsions

The influence of native and thermally (50–95 °C) denatured β-lactoglobulin (β-Lg) on the oxidative stability of surfactant-stabilized menhaden oil-in-water emulsions (pH 7.0) was evaluated. β-Lg (500 μg/g oil) heated at 95 °C for 30 min provided the best protection against lipid oxidation, inhibiting the formation of lipid hydroperoxides and thiobarbituric acid reactive substances (TBARS) by 87% and 88%, respectively, following 7 days of storage. The possible mechanisms of antioxidant activity of native and heated β-Lg were evaluated by measuring peroxyl radical scavenging and iron chelating capacities of the protein treatments, as well as reactive sulfhydryl concentrations and tryptophan fluorescence (a marker of protein conformation changes). The aforementioned in vitro assays only partially corroborated the results from the oxidizing emulsion system since β-Lg heated at 95 °C exhibited the lowest iron chelation capacity and free sulfhydryl concentration, yet displayed the highest peroxyl radical scavenging capacity and inhibition of lipid oxidation in oil-in-water emulsions of all treatments tested. The results of this study demonstrate the feasibility of proteins as a natural class of antioxidants in food emulsions, and further elucidate the possible mechanisms by which proteins inhibit lipid oxidation.     

Antioxidant properties of various solvent extracts of mulberry (Morus indica L.) leaves

The antioxidant properties and total phenolic contents of methanol, acetone and water extracts of mulberry (Morus indica L.) leaves were examined. Various experimental models including iron (III) reducing capacity, total antioxidant capacity, DPPH radical scavenging activity and in vitro inhibition of ferrous sulphate-induced oxidation of lipid system were used for characterization of antioxidant activity of extracts. The three extracts showed varying degrees of efficacy in each assay in a dose-dependent manner. Methanolic extract with the highest amount of total phenolics, was the most potent antioxidant in all the assays used. In addition, the effect of temperature (50 °C and 100 °C), pH (3, 5, 7, 9 and 11) and storage (5 °C) on the antioxidant activity of methanolic extract was investigated. The antioxidant activity of the extract remained unchanged at 50 °C and was maximum at neutral pH. The extract stored at 5 °C in the dark was stable for 30 days after which the antioxidant activity decreased (p ? 0.05) gradually. On the basis of the results obtained, mulberry leaves were found to serve as a potential source of natural antioxidants due to their marked antioxidant activity.     

Effects of caffeic acid and bovine serum albumin in reducing the rate of development of rancidity in oil-in-water and water-in-oil emulsions

The antioxidant properties of caffeic acid and bovine serum albumin in oil-in-water and water-in-oil emulsions were studied. Caffeic acid (5 mmol/kg emulsion) showed good antioxidant properties in both 30% sunflower oil-in-water (OW) and 20% water-in-sunflower oil emulsions (WO), pH 5.4, during storage at 50 °C. Although bovine serum albumin (BSA) (0.2%) had a slight antioxidant effect, the combination of caffeic acid and BSA showed a synergistic reduction in the rate of development of rancidity, with significant reductions in concentration of total volatiles, peroxide value (PV) and p-anisidine value (PA) for both emulsion types. The synergistic increase in stability of the OW and WO emulsions containing BSA and caffeic acid was 102.9% and 50.4% respectively based on total oxidation (TOTOX) values, which are calculated as 2PV + PA, with greater synergy calculated if based on formation of headspace volatiles. The OW emulsion was more susceptible to the development of headspace volatiles by oxidation than the WO emulsion, even though the degree of oxidation assessed by the TOTOX value was similar.     

Antioxidant activity of 1,4-dihydropyridine derivatives in β-carotene-methyl linoleate,sunflower oil and emulsions

The antioxidant activity of six synthetic 1,4-dihydropyridine (DHP) derivatives was tested in an azobis-amidinopropane dihydrochloride initiated β-carotene-methyl linoleate peroxidation model system. Radical scavenging activity of the derivatives was estimated by measuring their reactivity with stable N,N-diphenyl-N′-picrylhydrazyl radicals. The antioxidant activity of these different derivatives was also evaluated in traditional sunflower oil and its 20% oil-in-water emulsion in the dark at 60°C, as well as during storage of sunflower oil in the dark and with light exposure at ambient temperatures. The primary (conjugated diene hydroperoxides) and secondary (hexanal, pentanal) oxidation products were monitored during different periods of storage. Butylated hydroxytoluene was used as a standard antioxidant and different plant extracts were used for comparison of antioxidant activities throughout this study. All synthetic compounds showed antioxidant activity, while plant extracts acted as pro-oxidants upon light exposure. Three derivatives of DHP, without substitution at position 4, showed highest antioxidative activity in model system and sunflower oil and its emulsion in the dark at 60°C as well as during storage of sunflower oil in the dark and with light exposure at ambient temperature. The hydrophilic derivative of DHP with a phenyl group at position 4 showed high radical scavenging activity, and high antioxidant activity in sunflower oil-in-water emulsion, but was less active in model system and sunflower oil. The hydrophobic derivatives of DHP with a phenyl group at position 4 showed high antioxidant activity in model system but were the lowest active derivatives in sunflower oil and its emulsion. ©     

Determination of antioxidative potential of lichen Usnea ghattensis in vitro

The study was aimed at evaluating the antioxidant activities of extract of Usnea ghattensis. The antioxidant activity, reducing power, superoxide anion radical scavenging and free radical scavenging activities were studied. The antioxidant activity increased with the increasing amount of extracts (2-20 mg/ml) added to the linoleic acid emulsion. Lipid peroxidation upto 73.3% was inhibited by the extract of 20 mg/ml and 39.2% by α-tocopherol at the same concentration. These effects were statistically significant (r²=0.876,P<0.01) when compared with control. However, the extract had no significant effect on superoxide anion scavenging by the PMS/NADH-NBT method. Like antioxidant activity, the reducing power and free radical scavenging activity of extract depends on concentration and increasing with increased amount of sample. The reducing power and DPPH radical-scavenging activity of U. ghattensis extract were found greater than the BHA and BHT. The results obtained in the present study indicate that U. ghattensis is a potential source of natural antioxidant.     

Effect of processing on the antioxidant activity of millet grains

Millets are generally dehulled and subjected to a hydrothermal treatment before consumption, thus the hulls can be used as a potential source of antioxidants. Several millet grains, namely kodo, finger (Ravi), finger (local), proso, foxtail, little and pearl millet were studied. Antioxidant activities of phenolic extracts obtained from whole grains, as well as their corresponding dehulled and cooked grains and hulls were studied for their total phenolic content (TPC), radical scavenging capacity, and antioxidant activity in a β-carotene/linoleate emulsion. The phenolics present in whole grains were identified and quantified using HPLC and HPLC/MS and results were expressed as total for each of the phenolic groups. The TPC ranged from 2 to 112 μmol ferulic acid equivalents/g defatted meal. All varieties exhibited effective inhibition of 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl, peroxyl and superoxide radicals. Dehulling and cooking affected the TPC and radical scavenging and antioxidant activities of the grains, depending on the variety. In general, the antioxidant activity of phenolic extracts was in the order of hull > whole grain > dehulled grain > cooked dehulled grain With the exception of the two finger millet varieties, hulls of other millet grains had high TPC, thus demonstrating their superior antioxidant activity. Hydroxybenzoic acids, hydroxycinnamic acids and flavonoids in whole grains were identified as contributors to the observed effects. Therefore, dehulling of grain and hydrothermal treatments affect the phenolic content and antioxidant potential of millet grains.     

Antioxidant properties of dried product of ‘haba-nori’, an edible brown alga, Petalonia binghamiae (J. Agaradh) Vinogradova

Dried ‘haba-nori’ Petalonia binghamiae, a brown alga, is a traditional food in the fisheries towns in Japan. To determine the antioxidant properties of the dried P. binghamiae, assays for antioxidant activities, including ferrous-reducing power, ferrous ion chelating, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging and scavenging of a superoxide anion radical-generated by non-enzymatic system were tested in this study. A water extract solution contained total phenols at about 75 μmol phloroglucinol equivalents/g dry sample and showed strong antioxidant activities in the reducing power, DPPH radical and superoxide anion radical scavenging assays. The antioxidant activities were detected in high-molecular (>100 kDa), 10–30 kDa, and low-molecular (<5 kDa) fractions and were correlated with, not only phenolic compounds, but also brown compounds. The radical- scavenging activities were increased by heat treatment at 121 °C for 1 h. These results suggest that P. binghamiae is both a useful seafood and a healthy food with antioxidant activity.     

Antioxidative and functional properties of protein hydrolysate from defatted skipjack (Katsuwonous pelamis) roe

Antioxidative and functional properties of protein hydrolysate from defatted skipjack (Katsuwonous pelamis) roe, hydrolysed by Alcalase 2.4 L (RPH) with different degrees of hydrolysis (DH) at various concentrations were examined. As DH increased, the reduction of DPPH, ABTS radicals scavenging activities and reducing power were noticeable (p < 0.05). The increases in metal chelating activity and superoxide scavenging activity were attained with increasing DH (p < 0.05). However, chelating activity gradually decreased at DH above 30%. All activities except superoxide anion radical scavenging activity increased as the concentration of hydrolysate increased (p < 0.05). Hydrolysis using Alcalase could increase protein solubility to above 80% over a wide pH range (2–10). The highest emulsion ability index (EAI) and foam stability (FS) of hydrolysates were observed at low DH (5%) (p < 0.05). Concentrations of hydrolysates determined interfacial properties differently, depending on DH. The molecular weight distribution of RPH with 5%DH (RPH5) was determined using Sephadex G-75 column. Two major peaks with the molecular weight of 57.8 and 5.5 kDa were obtained. Fraction with MW of 5.5 had the strongest metal chelating activity and ABTS radical scavenging activity. The results reveal that protein hydrolysates from defatted skipjack roe could be used as food additives possessing both antioxidant activity and functional properties.     

Activity of natural carotenoid preparations against the autoxidative deterioration of sunflower oil-in-water emulsions

The activity of natural carotenoid extracts against the autoxidative deterioration of sunflower oil-in-water emulsions was estimated in terms of primary (lipid hydroperoxides) and secondary (“off flavour” volatiles) oxidation products. At a concentration of 2 g l⁻¹, the tested carotenoids did not inhibit the production of hydroperoxides but they significantly retarded the formation of volatile aldehydes during the storage of simple tween-stabilised emulsions at 30 °C. In another series of experiments, several carotenoid extracts (paprika, annatto and marigold preparations) containing mainly polar carotenoids, added at an active concentration of 1 g l⁻¹, exerted a strong activity against hydroperoxides and TBARs during the accelerated oxidation (60 °C) of homogenised protein-based emulsions. On the contrary, carotene preparations rich in hydrophobic α- and β-carotenes and lycopene did not significantly differ from the control emulsion. Therefore, the carotenoid structure modulated their antioxidant effect, while concentration and emulsion structure may also affect carotenoid activity in protein dispersed systems.     

Antioxidant activity of cichoric acid and alkamides from Echinacea purpurea, alone and in combination

The antioxidant activity of extracts of the stems, leaves, and roots of Echinacea purpurea was compared with the antioxidant activity of purified cichoric acid and alkamides, both constituents of Echinacea purpurea. The antioxidant activity was determined using different methods: effect on oxygen consumption rate of a peroxidating lipid emulsion, and scavenging of radicals, i.e. 2,2-diphenyl-1-picrylhydrazyl (DPPH), measured by two different techniques. The efficacy of the extracts in the reaction with DPPH correlated well with the amount of cichoric acid present in the various extracts. The alkamides alone showed no antioxidant activity in any of the tests. Alkamides present in the extract increased, however, the antioxidative effect of cichoric acid in the peroxidating lipid emulsion. The activity was further compared with that of rosmarinic acid, a well-characterised antioxidant, and the extracts as well as cichoric acid were found to be efficient scavengers of radicals with an activity comparable to that of rosmarinic acid. Cichoric acid was found to have a stoichiometric factor of 4.0 in scavenging DPPH and to react in a second-order reaction with DPPH with a rate constant of 40 l/mol/s at 25 °C in methanol.     

Effect of air-drying temperature on antioxidant capacity and stability of polyphenolic compounds in mulberry (Morus alba L.) leaves

The mulberry (Morus alba L.) leaf is a promising dietary source of antioxidants such as quercetin due to its relatively high content of that compound. We investigated effects of an air-drying process on the antioxidant capacity and stability of antioxidant polyphenolic compounds in mulberry leaves. Main compounds playing a central role in antioxidant activities in mulberry leaves are quercetin glycosides and chlorogenic acid. Raw mulberry leaves were air-dried at various temperatures, and antioxidant activity using DPPH radical scavenging assay and levels of antioxidant compounds were measured. DPPH radical scavenging activity and levels of polyphenolic compounds in mulberry leaves air-dried at 60 °C or below were not significantly different from those of freeze-dried mulberry leaves, whereas both values in mulberry leaves air-dried at 70 °C and over decreased significantly. These results indicate that strict temperature control is important in the production of mulberry leaf products to maintain antioxidant activity and levels of polyphenolic compounds.     

Optimisation of antioxidants extraction from soybeans fermented by Aspergillus oryzae

The extraction of antioxidant compounds from soybeans fermented with Aspergillusoryzae was optimised using a factorial design. A kinetic study of the total phenolic production and DPPH radical scavenging activity was first performed at the points selected in the factorial design. In both cases, the experimental profiles were fitted to a modified first-order kinetic model. To investigate the combined effects of temperature and solvent concentration on the extraction, the parameters obtained from the fitted kinetic models were used as response variables in a rotatable second-order design with quintuple replications in the centre of the experimental domain. The results obtained indicate that temperature had the most significant effect. The response surfaces show a maximum in the experimental domain studied. The optimum conditions for the extraction of total phenolic content were 65.3 °C and 73.1% ethanol, in which 56.2 mg of GAE/g were predicted. A scavenging activity of 81.6% DPPH radical was predicted at the optimum conditions of 61.6 °C and 60% ethanol.     

Antioxidant and radical scavenging activities of the pyroligneous acid from a mangrove plant, Rhizophora apiculata

Total phenolics content, free radical scavenging activity, reducing power, and antioxidant activity of the pyroligenous acid from a mangrove plant, Rhizophora apiculata were evaluated. Dichloromethane extraction of the raw pyroligneous acid successfully yield 2 extracts, i.e. concentrated pyroligneous acid (CPA) and concentrated pyroligneous acid extract (CPAE). Phenolic contents in CPAE and CPA, expressed as (±)-catechin equivalents/g of the sample were 5465 ± 367 mg and 2502 ± 152 mg, and expressed as gallic acid equivalents/g of the sample were 2919 ± 209 mg and 1348 ± 90 mg, respectively. CPAE exhibited superior free radical scavenging activity with EC₅₀ value = 0.1235 mg/ml, or 80.96% of free radical scavenging capability. The ferric reducing power of CPAE was approximately 3.7, 5.1, 6.1, and 21.3 times higher than that of ascorbic acid, BHA, BHT and alpha-tocopherol. In phosphomolybdenum assay, CPAE showed the greatest antioxidant efficacy (A₆₉₅ = 1.278) compared to those of CPA and different standards. In addition, the free radical scavenging activity, ferric reducing power and total antioxidative activity of CPAE and CPA showed positive correlation with their total phenolic content with R² values ranging from 0.9624 to 0.9979.     

In vitro determination of antioxidant activity of proteins from jellyfish Rhopilema esculentum

In this study, the antioxidant activity of proteins isolated from jellyfish, Rhopilema esculentum Kishinouye (R. esculentum), was determined by various antioxidant assays, including superoxide anion radical-scavenging, hydroxyl radical-scavenging, total antioxidant activity, reducing power and metal chelating activity. Butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), α-tocopherol, vitamin C and mannitol were used as standards in those various antioxidant activities. The crude protein (CP) and the protein fractions isolated by Sephadex chromatography, first peak (FP) and second peak (SP), had very low reductive power and metal chelating abilities compared to EDTA, but they showed strong scavenging effects on the superoxide anion radical, hydroxyl radical and varying total antioxidant activity. FP and SP exhibited stronger scavenging effects on the superoxide anion radical than BHA, BHT or α-tocopherol. The EC₅₀ values of FP and SP were 6.12 and 0.88 μg/ml, respectively, while values EC₅₀ of BHA, BHT and α-tocopherol were 31, 61 and 88 μg/ml, respectively. CP, FP and SP showed far higher hydroxyl radical-scavenging activities than did vitamin C or mannitol. The EC₅₀ values of CP, FP and SP were 48.76, 45.42 and 1.52 μg/ml, but EC₅₀ values of vitamin C and mannitol were 1907 and 4536 μg/ml, respectively. In a β-carotene–linoleate system, SP and CP showed antioxidant activity, but lower than BHA. Of the three samples, SP had the strongest antioxidant activity. So, SP may have a use as a possible supplement in the food and pharmaceutical industries.     

Quality and antioxidant properties of a reduced-sugar pomegranate juice jelly with an aqueous extract of pomegranate peels

In the present study, the production of a reduced-sugar pomegranate juice jelly supplemented with an aqueous extract of pomegranate peel (PE) is described. Influence of different carbohydrate polymers (guar (G), xanthan (X) and tragacanth (T) gums) on rheological properties was studied. Combination GXT presented the most similar rheological behaviour to commercial jelly. Jelly (J) and jelly with PE (JE) were stored at 4 °C over an 8 week period for physical, chemical, antioxidant, microbiological and sensory analysis. J and JE showed similar values for °Brix, colour and Aw, though the pH of JE was lower than J. Thiol and phenolic compounds were higher in JE than in J. Antioxidant activity (radical scavenging activity and autoxidation of linoleic acid) was higher in JE than in J at 0 weeks, and were decreasing with time. Pomegranate juice with additives was generally less accepted than J and JE.     

Evaluation of antioxidant activity of some plant extracts and their heat,pH and storage stability

In the present study, three plant foods, namely, drumstick leaves (Moringa oleifera), mint leaves (Mentha spicata) and carrot tuber (Daucus carota) were extracted with ethanol and analyzed for their antioxidant activity. The antioxidant activity of extracts was evaluated according to the amount of malonaldehyde (MDA) formed by the FeSO₄-induced oxidation of linoleic acid and a high PUFA oil (sunflower oil) at 37 °C in Trizma-buffer (pH 7.4). At a concentration of 1.5 mg/ml of linoleic acid ,the extracts from drumstick and carrot had a higher antioxidant activity (83% and 80%) than α-tocopherol (72%). In sunflower oil, the extracts from drumstick leaves and mint leaves were found to exhibit a similar activity( 46% and 44%). The extract from drumstick exhibited the highest activity in both lipid systems. In addition, the stability of extracts to pH (4 and 9) and temperature (100 °C, 15 min) was investigated. The antioxidant activity of the extracts from mint leaves and carrot was higher at pH 9 than pH 4, while that of drumstick extract remained the same under both pH conditions. The extract from carrot was more heat-stable than other extracts. The three extracts stored in the dark at 5 and 25 °C after a 15 day period did not show any significant change (p ? 0.05) in their antioxidant activity. These data indicate that selected plant extracts are potential sources of dietary antioxidants.     

Autolysis-assisted production of fish protein hydrolysates with antioxidant properties from Pacific hake (Merluccius productus)

Fish protein hydrolysates (FPH) with antioxidative properties were prepared using Pacific hake fish with high endogenous proteolytic activity from Kudoa paniformis parasitic infection. Infection level of ∼10⁷K. paniformis spores/g fish mince or higher yielded FPH with high antioxidant potential by autolysis and/or Validase^® BNP or Flavourzyme^® 500L. Autolyzing fish mince containing 30 × 10⁶ spores/g for 1 h at 52 °C and pH 5.50 produced FPH (named E-1h) with Trolox equivalent antioxidant capacity (TEAC) in the 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulphonic acid) radical assay of 262 ± 2 μmol/g and oxygen radical absorbing capacity (ORAC) of 225 ± 17 μmol Trolox equivalents/g freeze-dried sample. E-1h FPH also exhibited a marked concentration-dependent scavenging activity on 1,1-diphenyl-2-picrylhydrazyl radical. Antioxidant activity of E-1h FPH was higher (p < 0.05) than BHA and α-tocopherol in a linoleic acid peroxidation system over prolonged storage (∼162 h). Antioxidative FPH from Pacific hake may be useful ingredients in food and nutraceutical applications.     

Antioxidant activity of ethanolic extract of Cortex fraxini and use in peanut oil

Cortex fraxini was extracted with 95% ethanol to obtain a crude antioxidant extract. The antioxidant activity was evaluated using the linoleic acid peroxidation method and the free radical scavenging assays, namely 2,2′-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl radicals. Cortex fraxini extract (CFE) showed high inhibition of peroxidation of linoleic acid when compared with butylated hydroxytoluene (BHT). CFE also exhibited excellent scavenging activity on DPPH and hydroxyl radicals. Total antioxidant activity was measured by the reduction of Mo(VI) to Mo(V) by the extract, and subsequent formation of a green phosphate/Mo(V) complex at acid pH. CFE had significant total antioxidant activity and the effects were increased with increasing reaction time. The total phenolic content of the sample, analyzed by using Folin–Ciocalteu’s reagent, was 91.33 mg/g dry weight expressed as pyrocatechol equivalents. Then the suitability of CFE as an antioxidant was determined in peanut oil, and the decrease of lipid oxidation was monitored using thiobarbituric acid-reactive substances (TBARS) assay. CFE treatment significantly (P < 0.05) reduced lipid oxidation in peanut oil compared to the control. No significant differences (P = 0.05) in lipid oxidation were detected between CFE antioxidant and BHT antioxidant samples.     

Antioxidant and free radical scavenging potential of Achillea santolina extracts

The antioxidative activities of hydroalcoholic extract of Achillea santolina were investigated employing various established in vitro systems including total antioxidant activity in linoleic acid emulsion system, 1,1-diphenyl-2-picrylhydrazyl (DPPH), superoxide and hydroxyl radicals scavenging, reducing power, and inhibitory effect on protein oxidation as well as the inhibition of Fe²⁺/ascorbate induced lipid peroxidation in rat liver homogenate. Total phenolic and flavonoid content of A. santolina extract (ASE) was also determined by a colorimetric method. The results revealed that ASE has notable inhibitory activity on peroxides formation in linoleic acid emulsion system along with concentration-dependent quenching of DPPH and superoxide radicals. Furthermore, the extract showed both nonsite-specific (Fe²⁺ + H₂O₂ + EDTA) and site-specific (Fe²⁺ + H₂O₂) hydroxyl radical scavenging suggesting potent hydroxyl radical scavenging and chelating ability for iron ions in deoxyribose degradation model. A linear correlation between ASE and the reducing power was also observed (r² = 0.9981). ASE prevents thiobarbituric acid reactive substances formation in Fe²⁺/ascorbate induced lipid peroxidation in rat liver tissue in a dose-dependent manner. Moreover, free radical induced protein oxidation was suppressed significantly by the addition of ASE over a range of concentration. These results clearly demonstrated that A. santolina extract possess a marked antioxidant activity.