Simple assessment of radical scavenging capacity of beverages

The radical-scavenging antioxidants play an important role against oxidative stress in the defense system in vivo. The beneficial effects of antioxidants contained in foods and beverages have been well-accepted, and their antioxidant capacity has been assessed by various methods. In the present study, a simple method is proposed in which the total radical scavenging capacity is assessed from the bleaching of pyranine and pyrogallol red induced by free radicals generated from azo initiator. The total content of antioxidants contained in red wine, green tea, and cassis drink and their reactivities toward peroxyl radicals were measured from the lag phase and rate of bleaching using pyranine and pyrogallol red as a probe, respectively. It was found that this method to follow the bleaching of two probes by visible light spectrophotometer is convenient and applicable for assessment of total radical scavenging capacity of both content and activity of the antioxidants contained in beverages.     

Novel fluorometric assay for hydroxyl radical scavenging capacity (HOSC) estimation

A novel fluorometric method was developed and validated for hydroxyl radical scavenging capacity (HOSC) estimation using fluorescein as the probe. A constant flux of pure hydroxyl radical is generated under physiological pH using a Fenton-like Fe3+/H2O2 reaction. The generation of pure hydroxyl radicals under the experimental conditions was evaluated and confirmed using electron spin resonance with DMPO spin-trapping measurements. The hydroxyl radical scavenging capacity of a selected antioxidant sample is quantified by measuring the area under the fluorescence decay curve with or without the presence of the antioxidant and expressed as Trolox equivalents per unit of the antioxidant. The assay may be performed using a plate reader with a fluorescence detector for high-throughput measurements. The assay was validated for linearity, precision, accuracy, reproducibility, and its correlation with a popular peroxyl radical scavenging capacity assay using selected pure antioxidant compounds and botanical extracts. This method may provide researchers in the food, nutrition, and medical fields an easy to use protocol to evaluate free radical scavenging capacity of pure antioxidants and natural extracts in vitro against the very reactive hydroxyl radical, which may be linked to numerous degenerative diseases and conditions.     

Antioxidant activity of glyceollins derived from soybean elicited with Aspergillus sojae

The extract of soybean exposed to biotic elicitors such as food-grade fungus is known to have antioxidant activity. Glyceollins were major bioactive compounds present in soybean elicited by fungi and shown to have antifungal and anticancer activities. The purpose of present study was to evaluate the antioxidant activities of glyceollins by measuring ferric reducing antioxidant power (FRAP), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, singlet oxygen quenching, 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging, hydroxyl radical scavenging activity, and lipid peroxidation inhibition. In addition, the antioxidant potential of glyceollins were measured by a fluorescent probe, 2',7'-dichlorofluorescin diacetate (DCFDA), and dihydroethidium (DHE) in mouse hepatoma hepa1c1c7 cells in which they were insulted with H2O2 to generate reactive oxygen species (ROS). Glyceollins showed a strong reducing power and inhibited lipid peroxidation, with significant scavenging activities of radicals including singlet oxygen, superoxide anion, ABTS, and DPPH. We also found that glyceollins significantly suppressed H2O2-induced ROS production in hepa1c1c7 cells. Therefore, glyceollins deserve further study as natural antioxidants and nutraceuticals.     

Antioxidant capacity and other bioactivities of the freeze-dried Amazonian palm berry, Euterpe oleraceae mart. (acai)

The fruit of Euterpe oleraceae, commonly known as acai, has been demonstrated to exhibit significantly high antioxidant capacity in vitro, especially for superoxide and peroxyl scavenging, and, therefore, may have possible health benefits. In this study, the antioxidant capacities of freeze-dried acai fruit pulp/skin powder (OptiAcai) were evaluated by different assays with various free radical sources. It was found to have exceptional activity against superoxide in the superoxide scavenging (SOD) assay, the highest of any food reported to date against the peroxyl radical as measured by the oxygen radical absorbance capacity assay with fluorescein as the fluorescent probe (ORACFL), and mild activity against both the peroxynitrite and hydroxyl radical by the peroxynitrite averting capacity (NORAC) and hydroxyl radical averting capacity (HORAC) assays, respectively. The SOD of acai was 1614 units/g, an extremely high scavenging capacity for O2*-, by far the highest of any fruit or vegetable tested to date. Total phenolics were also tested as comparison. In the total antioxidant (TAO) assay, antioxidants in acai were differentiated into "slow-acting" and "fast-acting" components. An assay measuring inhibition of reactive oxygen species (ROS) formation in freshly purified human neutrophils showed that antioxidants in acai are able to enter human cells in a fully functional form and to perform an oxygen quenching function at very low doses. Furthermore, other bioactivities related to anti-inflammation and immune functions were also investigated. Acai was found to be a potential cyclooxygenase (COX)-1 and COX-2 inhibitor. It also showed a weak effect on lipopolysaccharide (LPS)-induced nitric oxide but no effect on either lymphocyte proliferation and phagocytic capacity.     

High-throughput quantitation of peroxyl radical scavenging capacity in bulk oils

Autoxidation of methyl linoleate (8:2 mixture with decane, 37 degrees C) was induced by 2,2'-azobis(2,4-dimethylvaleronitrile) (AMVN, 17.7 mM) and the kinetics of oxygen consumption monitored using a 96-well microplate coated with an oxygen-sensitive fluorescence probe, a ruthenium dye, embedded in a silicone matrix at the bottom of the microplate. The probe does not participate in the reaction; instead, its fluorescence intensity is inversely proportional to the solution oxygen concentration as it changes during oxidation. In the absence of antioxidants, the oxidation rate has a linear relationship with the square root of the initiator concentrations. This is in agreement with theoretical autoxidation kinetics equations. In the presence of tocopherol-type antioxidants, a sharp lag phase appears. The quantitation of the antioxidant capacity is achieved using the area under the curve (AUC) approach. The assay has a 2 h running time, a linearity range from 1.56 to 18.7 microM (Trolox), and a limit of quantitation at 2.7 microM Trolox equivalency. The peroxyl radical scavenging capacities of several cold-pressed and organically grown plant seed oils were quantified along with the tocopherol concentrations of the oils. Tocopherols contribute only a fraction of the peroxyl radical scavenging capacity of the oils, and there is poor correlation between total tocopherol concentrations and radical scavenging capacity, suggesting that the antioxidant capacity of oils is due not only to tocopherols but also to other lipid-soluble antioxidants.     

Assays for hydrophilic and lipophilic antioxidant capacity (oxygen radical absorbance capacity (ORAC(FL))) of plasma and other biological and food samples

Methods are described for the extraction and analysis of hydrophilic and lipophilic antioxidants, using modifications of the oxygen radical absorbing capacity (ORAC(FL)) procedure. These methods provide, for the first time, the ability to obtain a measure of "total antioxidant capacity" in the protein free plasma, using the same peroxyl radical generator for both lipophilic and hydrophilic antioxidants. Separation of the lipophilic and hydrophilic antioxidant fractions from plasma was accomplished by extracting with hexane after adding water and ethanol to the plasma (hexane/plasma/ethanol/water, 4:1:2:1, v/v). Lipophilic and hydrophilic antioxidants were efficiently partitioned between hexane and aqueous solvents. Conditions for controlling temperature effects and decreasing assay variability using fluorescein as the fluorescent probe were validated in different laboratories. Incubation (37 degrees C for at least 30 min) of the buffer to which AAPH was dissolved was critical in decreasing assay variability. Lipophilic antioxidants represented 33.1 +/- 1.5 and 38.2 +/- 1.9% of the total antioxidant capacity of the protein free plasma in two independent studies of 6 and 10 subjects, respectively. Methods are described for application of the assay techniques to other types of biological and food samples.     

Novel fluorometric assay for hydroxyl radical prevention capacity using fluorescein as the probe

A novel fluorometric method has been developed to evaluate hydroxyl radical prevention capacity using fluorescein (FL) as the probe. The hydroxyl radical is generated by a Co(II)-mediated Fenton-like reaction, and the hydroxyl radical formation under the experimental condition is indirectly confirmed by the hydroxylation of p-hydroxybenzoic acid. The fluorescence decay curve of FL is monitored in the absence or presence of antioxidant, the area under the fluorescence decay curve (AUC) is integrated, and the net AUC, which is an index of the hydroxyl radical prevention capacity, is calculated by subtracting the AUC of the blank from that of the antioxidant. Gallic acid is chosen as a reference standard, and the activity of sample is expressed as gallic acid equivalents. The method is rigorously validated through linearity, precision, accuracy, and ruggedness. A wide range of phenolic antioxidants is analyzed, and the hydroxyl radical prevention capacity is mainly due to the metal-chelating capability of the compounds.     

The chemistry behind antioxidant capacity assays

This review summarizes the multifaceted aspects of antioxidants and the basic kinetic models of inhibited autoxidation and analyzes the chemical principles of antioxidant capacity assays. Depending upon the reactions involved, these assays can roughly be classified into two types: assays based on hydrogen atom transfer (HAT) reactions and assays based on electron transfer (ET). The majority of HAT-based assays apply a competitive reaction scheme, in which antioxidant and substrate compete for thermally generated peroxyl radicals through the decomposition of azo compounds. These assays include inhibition of induced low-density lipoprotein autoxidation, oxygen radical absorbance capacity (ORAC), total radical trapping antioxidant parameter (TRAP), and crocin bleaching assays. ET-based assays measure the capacity of an antioxidant in the reduction of an oxidant, which changes color when reduced. The degree of color change is correlated with the sample's antioxidant concentrations. ET-based assays include the total phenols assay by Folin-Ciocalteu reagent (FCR), Trolox equivalence antioxidant capacity (TEAC), ferric ion reducing antioxidant power (FRAP), "total antioxidant potential" assay using a Cu(II) complex as an oxidant, and DPPH. In addition, other assays intended to measure a sample's scavenging capacity of biologically relevant oxidants such as singlet oxygen, superoxide anion, peroxynitrite, and hydroxyl radical are also summarized. On the basis of this analysis, it is suggested that the total phenols assay by FCR be used to quantify an antioxidant's reducing capacity and the ORAC assay to quantify peroxyl radical scavenging capacity. To comprehensively study different aspects of antioxidants, validated and specific assays are needed in addition to these two commonly accepted assays.     

Novel mechanism of modulating natural antioxidants in functional foods: involvement of plant growth promoting Rhizobacteria NRRL B-30488

The significance of plant growth-promoting rhizobacteria (PGPR) mediated increase in antioxidant potential in vegetables is yet unknown. The plant growth-promoting bacterium Bacillus lentimorbus NRRL B-30488 (B-30488) mediated induction of dietary antioxidant in vegetables ( Trigonella foenum-graecum, Lactuca sativa, Spinacia oleracea, and Daucus carota) and fruit ( Citrus sinensis) after minimal processing (fresh, boiled, and frozen) was tested by estimating the total phenol content, level of antioxidant enzymes, and 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide scavenging activities along with integral radical scavenging capacity by photochemiluminescence assay and inhibition of lipid peroxidation. Minimal processing of vegetables showed that T. foenum-graecum had the highest phenol content in B-30488-treated plants followed by L. sativa, D. carota, and S. oleracea. Thermally treated vegetables T. foenum-graecum (26-114.5 GAE microg mg (-1)) had an exceptionally high total phenolic content, followed by D. carota (25.27-101.32 GAE microg mg (-1)), L. sativa (23.22-101.10 GAE microg mg (-1)), and S. oleracea (21.87-87.57 GAE microg mg (-1)). Among the vegetables and fruit used in this study for enzymatic estimation, induction of antioxidant enzymes, namely, polyphenol oxidase (PPO), ascorbate peroxidase (APX), catalase (CAT), and superoxidase dismutase (SOD), was observed in edible parts of T. foenum-graecum, L. sativa, S. oleracea, and D. carota, after inoculation with B-30488. The scavenging capacity of the vegetables treated with B-30488 against DPPH and superoxide anion radical activity was found to be significantly high as compared to nontreated control. Mild food processing had no adverse effect on radical scavenging capacity. Photochemiluminescence also ascertains the above findings. The ability of the plant extracts to protect against lipid peroxidation and its ability to prevent oxidation of reduced glutathione (GSH) was measured in rat liver homogenate, and the results suggested that the inoculated plant exhibited better activity in all of the screened plants. Significant increases in shoot length, root length, and dry weight, averaging 164, 132, and 135% in T. foenum-graecum, 174, 141, and 156% in L. sativa, 129, 141, and 59%, in S. oleracea, and 125, 146, and 42% in D. carota, respectively, over untreated controls, were attained in greenhouse trials. To the best of the authors' knowledge, this is the first report of PGPR-mediated induction of antioxidant enzyme activity (PPO, APX, CAT, and SOD) along with the antioxidant activity of the extracts in both in vitro (DPPH radical scavenging and superoxide scavenging) and ex vivo conditions using the rat liver tissue (percent inhibition of lipid peroxidation and prevention of oxidation of GSH) and phenolic content. The results demonstrate the PGPR-mediated induction of antioxidant level in vegetables and fruit controls oxidative damage even after minimal processing and thus is indicative of its potential as a viable substitute of synthetic antioxidants.     

Novel low-density lipoprotein (LDL) oxidation model: antioxidant capacity for the inhibition of LDL oxidation

A novel model of peroxyl radical initiated low-density lipoprotein (LDL) oxidation (LDL oxidation model for antioxidant capacity, or LOMAC) was developed to assess the free radical scavenging capacity of antioxidants and the extracts of natural products. A water-soluble free radical initiator, 2,2'-azobis(amidinopropane) dihydrochloride, was used at physiological temperature (37 degrees C) to generate peroxyl radicals to catalyze lipid oxidation of LDL isolated from human plasma samples. Headspace hexanal, a major decomposition product of LDL oxidation, was measured by a headspace gas chromatograph as an indicator of antioxidant capacity of different concentrations of pure antioxidants (vitamins C and E) and the extracts of natural products (fresh apple phytochemical extracts). All vitamin C and E and apple extract concentrations tested resulted in increasing partial suppression and delay of LDL oxidation. On the basis of the median effective dose (EC(50)) calculated for each compound or extract tested, the LOMAC value of 100 g of apple against LDL oxidation was equivalent to 1470 mg of vitamin E or to 402 mg of vitamin C. This study shows that the LOMAC assay can be routinely used to analyze or screen antioxidants or phytochemical extracts against LDL oxidation to prevent cardiovascular disease. The food-specific LOMAC values will be very useful as a new alternative biomarker for future epidemiological studies of cardiovascular disease.     

Antioxidant profile of red wines evaluated by total antioxidant capacity, scavenger activity, and biomarkers of oxidative stress methodologies

The study of the antioxidant capacity of foodstuffs requires the use of diverse determination methods to gain a wider picture of their multiple effects. The aim of this work was to evaluate the "antioxidant profile" of red wines applying TAC (total antioxidant capacity) methods: 2,2'-azinobis(3-ethyl-benzthiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl, N,N-dimethyl-p-phenylenediamine dihydrochloride, oxygen radical absorbance capacity, ferric reducing/antioxidant power, hydroxyl and superoxide radical scavenger activities, and biomarkers of oxidative stress methods such as lipid peroxidation inhibition and inhibition of damage to DNA. Furthermore, levels of total polyphenols (TPP) of wines were also evaluated. Three bottles of 107 different Spanish red wines (total samples 321), made from different grape varieties, aging processes, and vintages, were analyzed. The validation of TAC methods, the first step in this work, provided a good linearity, proportionality, and low detection limits. Among these methods, the ABTS was the most satisfactory for its rapidity, cost, and precision. All wines showed an important capacity to scavenge hydroxyl radicals and were capable of blocking superoxide radicals but with 10 times lower intensity. Wines also showed important protective action on biomarkers of oxidative stress; they were much more active to inhibit lipid peroxidation than DNA oxidation. Few statistically significant correlations among levels of TPP and antioxidant properties of wines were detected. Furthermore, values of these correlations were very low.     

Evaluation of the antioxidant capacity of individual phenolic compounds in virgin olive oil

Virgin olive oil has a high resistance to oxidative deterioration due to its tryacylglycerol composition low in polyunsaturated fatty acids and due to the presence of a group of phenolic antioxidants composed mainly of polyphenols and tocopherols. We isolated several phenolic compounds of extra virgin olive oil (phenyl-ethyl alcohols, lignans, and secoiridoids) by semipreparative high-performance liquid chromatography (HPLC) and identified them using ultraviolet, atmospheric pressure chemical ionization, and electrospray ionization MS detection. The purity of these extracts was confirmed by analytical HPLC using two different gradients. Finally, the antioxidant capacity of the isolated compounds was evaluated by measuring the radical scavenging effect on 1,1-diphenyl-2-picrylhydrazyl radical, by accelerated oxidation in a lipid model system (OSI, oxidative stability instrument), and by an electrochemical method.     

Antioxidant Capacity of Newly Developed Pigmented Rice Cultivars in Korea

The antioxidant capacity of newly developed and highly popular pigmented rice cultivars (black rice, Galsaekchalmi, Jeoktomi, Hongchalmi, and Nogwonmi) in South Korea was analyzed. The rice grains were ground into powder, extracted with 70% ethanol, filtered, and concentrated with a rotary evaporator. The samples were analyzed for phenolic, flavonoid, and phytic acid contents, free radical scavenging activity, reducing power, ferrous ion chelating ability, lipid peroxidation inhibition, and superoxide dismutase‐like activity. The ethanolic extracts from pigmented rice cultivars showed greater antioxidant activity than that of the normal white rice. The black rice exhibited the highest free radical scavenging activity, ferrous chelating ability, and total phenolic and flavonoid contents. The reducing power and phytic acid content were found to be highest in Hongchalmi cultivar. The inhibition of lipid peroxidation was markedly higher in Jeoktomi compared with the other rice samples. The Nogwonmi rice showed the lowest antioxidant activity among the pigmented varieties analyzed. These findings provide valuable information on the antioxidant potential of newly developed pigmented rice varieties and may assist plant breeders in the selection of cultivars for the development of new lines of rice with enhanced functional quality.     

Studies on the antioxidant activity of pomegranate (Punica granatum) peel and seed extracts using in vitro models.

Antioxidant-rich fractions were extracted from pomegranate (Punica granatum) peels and seeds using ethyl acetate, methanol, and water. The extracts were screened for their potential as antioxidants using various in vitro models, such as beta-carotene-linoleate and 1,1-diphenyl-2-picryl hydrazyl (DPPH) model systems. The methanol extract of peels showed 83 and 81% antioxidant activity at 50 ppm using the beta-carotene-linoleate and DPPH model systems, respectively. Similarly, the methanol extract of seeds showed 22.6 and 23.2% antioxidant activity at 100 ppm using the beta-carotene-linoleate and DPPH model systems, respectively. As the methanol extract of pomegranate peel showed the highest antioxidant activity among all of the extracts, it was selected for testing of its effect on lipid peroxidation, hydroxyl radical scavenging activity, and human low-density lipoprotein (LDL) oxidation. The methanol extract showed 56, 58, and 93.7% inhibition using the thiobarbituric acid method, hydroxyl radical scavenging activity, and LDL oxidation, respectively, at 100 ppm. This is the first report on the antioxidant properties of the extracts from pomegranate peel and seeds. Owing to this property, the studies can be further extended to exploit them for their possible application for the preservation of food products as well as their use as health supplements and neutraceuticals.     

Scavenging capacity of berry crops on superoxide radicals, hydrogen peroxide, hydroxyl radicals, and singlet oxygen

The antioxidant activities against superoxide radicals (O(2)(*)(-)), hydrogen peroxide (H(2)O(2)), hydroxyl radicals (OH(*)), and singlet oxygen ('O(2)) was evaluated in fruit juice from different cultivars of thornless blackberries (Rubus sp.), blueberries (Vaccinium spp.), cranberries (Vaccinium macrocarpon Aiton), raspberries (Rubus idaeus L. and Rubus occidentalis L.), and strawberries (Fragaria x ananassa Duch.). Among the different cultivars, juice of 'Hull Thornless' blackberry, 'Earliglow' strawberry, 'Early Black' cranberry, 'Jewel' raspberry, and 'Elliot' blueberry had the highest antioxidant capacity against superoxide radicals (O(2)(*)(-)), hydrogen peroxide (H(2)O(2)), hydroxyl radicals (OH(*)), and singlet oxygen ('O(2)). In general, blackberries had the highest antioxidant capacity inhibition of O(2)(*)(-), H(2)O(2), and OH(*). Strawberry was second best in the antioxidant capacity assay for these same free radicals. With regard to 'O(2) scavenging activity, strawberry had the highest value, while blackberry was second. Cranberries had the lowest inhibition of H(2)O(2) activity. Meanwhile, blueberries had the lowest antioxidant capacity against OH(*) and 'O(2). There were interesting and marked differences among the different antioxidants in their abilities to scavenge different reactive oxygen species. beta-Carotene had by far the highest scavenging activity against 'O(2) but had absolutely no effect on H(2)O(2). Ascorbic acid was the best at inhibiting H(2)O(2) free radical activity. For OH(*), there was a wide range of scavenging capacities from a high of 15.3% with alpha-tocopherol to a low of 0.88% with ascorbic acid. Glutathione had higher O(2)(*)(-) scavenging capacity compared to the other antioxidants.     

Antioxidant activity of knotwood extractives and phenolic compounds of selected tree species

The antioxidant potency and the radical scavenging capacity of superoxide and peroxyl radicals were assessed for 13 hydrophilic knotwood extracts of commercially important wood species, or fractions thereof, as well as for five pure wood-derived lignans and the flavonoid taxifolin. The chemical composition of the knotwood extracts was determined by gas chromatography combined with mass spectrometry. Most of the investigated wood species were rich in hydrophilic extractives (10-20% of the dry wood) with one or a few compounds dominating in each extract. All extracts had a high antioxidative potency and/or radical scavenging capacity as compared to the well-known antioxidants Trolox and butylated hydroxyanisole. The pure wood-derived lignans and taxifolin also had a high antioxidative potency and/or radical scavenging capacity. However, the antioxidant potency and/or radical scavenging capacity of several of the hydrophilic knotwood extracts were higher than that of the dominating compounds in pure form.     

Antioxidant activities of grape (Vitis vinifera) pomace extracts

Antioxidant-rich fractions were extracted from grape (Vitis vinifera) pomace using ethyl acetate, methanol, and water. The extracts were screened for their potential as antioxidants in different models. The ethyl acetate, methanol, and water extracts showed 76, 87.1, and 21.7% antioxidant activities at 100 ppm, respectively, using the 1,1-diphenyl-2-picrylhydrazyl model system. As the methanol extract of grape pomace showed maximum antioxidant activity among all of the extracts, it was selected to determine its effect on lipid peroxidation, hydroxyl radical scavenging activity, and human low-density lipoprotein (LDL) oxidation. The methanol extract showed 71.7, 73.6, and 91.2% inhibition using the thiobarbituric acid method, hydroxyl radical scavenging activity, and LDL oxidation, respectively, at 200 ppm. Treatment of albino rats of the Wistar strain with a single dose of CCl(4) at 1.25 mL/kg of body weight decreases the activities of catalase, superoxide dismutase (SOD), and peroxidase by 81, 49, and 89%, respectively, whereas the lipid peroxidation value increased nearly 3-fold. Pretreatment of the rats with the methanolic extract of grape pomace at 50 mg/kg (in terms of catechin equivalents) followed by CCl(4) treatment causes restoration of catalase, SOD, and peroxidase by 43.6, 73.2, and 54%, respectively, as compared with control, whereas lipid peroxidation was restored to values comparable with the control. Histopathological studies of the liver of different groups also support the protective effects exhibited by the methanol extract of grape pomace by restoring the normal hepatic architecture. Owing to this property, the studies on grape pomace can be further extended to exploit its possible application for the preservation of food products as well as a health supplement and neutraceutical.     

Literature data may underestimate the actual antioxidant capacity of cereals

Several recent articles have reported a significant antioxidant capacity of cereal products, determined in methanolic and ethanolic extracts. The aim of this work was to conduct an assessment of the antioxidant capacity of cereals using both chemical and in vitro digestive enzymatic extraction of antioxidants. Ferric reducing power (FRAP) and free radical scavenging capacity (DPPH) methods were used to determine the antioxidant capacity in wheat flour, bread, raw and boiled rice, wheat bran, and oat bran. The most efficient antioxidant extraction was achieved by using successively acidic methanol/water (50:50 v/v, pH 2) and acetone/water (70:30 v/v). The antioxidant capacity in these extracts ranged from 1.1 to 4.4 micromol Trolox/g dw. A significant amount of hydrolyzable phenolics with a high antioxidant capacity (from 5 to 108 micromol Trolox/g dw) was found in the residues of this aqueous-organic extraction. The antioxidant capacities of these nonextractable polyphenols are usually ignored in the literature, although they may have an antioxidant role in the gastrointestinal tract, especially after colonic fermentation, and may be fermentated to active metabolites. On the other hand, in vitro digestive enzymatic extracts obtained by enzymatic treatments that mimic conditions in the gastrointestinal tract showed that the amount of antioxidants released by the cereal matrix into the human intestine may be higher than the one that can be expected from measurements in the usual aqueous-organic extracts.     

Antioxidant activity of methanol extracts obtained from Plantago species

Antioxidative activities of methanol extracts from five Plantago species (P. afra, P. coronopus, P. lagopus, P. lanceolata, and P. serraria) were characterized by the DPPH scavenging test and the inhibition of Fe2+/ascorbate-induced lipid peroxidation on bovine brain liposomes. All extracts showed antioxidant activity in both methods. Whereas P. serraria exhibited the strongest activity as a DPPH scavenger, P. lanceolata and P. serraria were found to be the most active in the lipid peroxidation inhibition assay. The extracts were investigated regarding their composition by different colorimetric techniques, such as the content of total phenolic compounds by the Folin-Ciocalteu assay, flavonoids by AlCl3 reagent, phenylpropanoid glycosides (PPGs) by Arnow reagent, and iridoids by Trim-Hill assay. A high correlation was found between the scavenging potency and the total phenolic and phenylpropanoid content of the extracts but not between the lipid peroxidation potency and the extract composition. P. serraria is presented as a possible new source of natural antioxidants.     

Effect of foliar application of selenium on the antioxidant activity of aqueous and ethanolic extracts of selenium-enriched rice

Selenium fertilizer was foliar applied to determine the effects of antioxidant activity of selenium-enriched rice assessed by alpha,alpha-diphenyl-beta-picylhydrazyl (DPPH) radical scavenging and the ferric thiocyanate (FTC) method. Results showed that selenium concentration in rice was significantly enhanced dose dependently. Aqueous or ethanolic extracts of rice displayed significantly higher antioxidant activity against lipid peroxidation. The activities of aqueous extracts were significantly higher than those of ethanolic extracts and increased with the increasing selenium concentration in rice. The DPPH assay showed that the kinetic behaviors of aqueous extracts were complex and slow, while ethanolic extracts reacted quickly with DPPH radical. Aqueous extracts of rice exhibited higher antiradical efficiencies than ethanolic extracts, and rice (1.275 mg Se kg(-)(1)) presented the lowest EC(50) values of 533.46 +/- 0.58 microg mL(-)(1). As compared to rice extracts, all of the reference antioxidants showed more than 4-fold antiradical efficiencies than rice extracts. This radical scavenging activity was significantly correlated with selenium concentrations in rice (R = 0.862, p < 0.05), while ethanolic extracts were inversely correlated with selenium concentration in rice.