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1.
Hydroxyl radical scavenging capacity estimation for lipophilic antioxidants is a challenge due to their poor solubility in aqueous radical generating and measuring systems. In this study, an electron spin resonance (ESR) method was developed and validated for its application in estimating the relative hydroxyl radical (HO*) scavenging capacity for lipophilic antioxidants under physiological pH using a Fenton Fe2+/H2O2 system for radical generation and acetonitrile as a solvent. The Fenton Fe2+/H2O2 system generates a constant flux of pure HO* under the assay conditions. The method was validated by linearity, precision, and reproducibility using selected known lipophilic antioxidants including alpha-tocopherol, lutein, beta-carotene, and BHT. The potential effects of commonly used water-miscible and water-immiscible organic solvents on the Fenton Fe2+/H2O2 HO* generating system as well as their possible interactions with the fluorescent and spectroscopic probes were also reported. In addition, the limitation of the ESR assay was described.  相似文献   

2.
The antioxidant activity of extracts from bark and heartwood of Acacia confusa was evaluated by various antioxidant assays, including free radical and superoxide radical scavenging assays and lipid peroxidation assay as well as hydroxyl radical-induced DNA strand scission assay. In addition, an ex vivo antioxidant assay using a flow cytometric technique was also employed in this study. The results indicate that both bark and heartwood extracts clearly have strong antioxidant effects. Similar inhibitory activities for each test sample were found for both 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical generation and lipid peroxidation. As for the superoxide radical scavenging activity, the heartwood extract was more effective than the bark extract. Furthermore, the heartwood extract protected PhiX174 supercoiled DNA against strand scission induced by ultraviolet photolysis of H2O2, and it reduced the amounts of intracellular hydrogen peroxide, a reactive oxygen species, when it was co-incubated with human promyelocytic leukemia (HL-60) cells under oxidative stress.  相似文献   

3.
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.  相似文献   

4.
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.  相似文献   

5.
This paper reports the use of spectrophotometric and voltammetric methods for the determination of the antioxidant capacity of buckwheat and its products originated from a technological line of a buckwheat roasted groats producer. 80% methanol extracts from raw and roasted buckwheat and groats and hulls obtained from roasted buckwheat were used. The spectrophotometric methods included (1) free radical scavenging activities of the extracts against ABTS*+ radical cation (TEAC) and 2,2-diphenyl-1-picrylhydrazyl radical (DPPH RSA) and (2) determination of reducing capacity by the means of Folin-Ciocalteu reagent (FCR) application. The radical scavenging activities of the extracts were also investigated using a voltammetric assay. Moreover, the flavonoids profiles of the studied materials were provided. Buckwheat roasting caused a decrease in TEAC, DPPH RSA, and FCR reducing capacity by 70%. The lowest TEAC, DPPH RSA, and FCR reducing capacities were noted for roasted groats. Both DPPH RSA and TEAC methods were highly positively correlated with the FCR reducing capacity assay (r = 0.98 and r = 0.99). Cyclic voltammograms of analyzed buckwheat extracts were useful for evaluation of the antioxidant capacity. The total charge below the anodic current waveform was correlated with the data obtained by TEAC (r = 0.770), DPPH RSA (r = 0.88), and FCR reducing capacity (r = 0.81). The changes in the antioxidant capacity of buckwheat and its products followed the changes in flavonoids composition. In particular, the concentration of flavonoids was related to measurements by cyclic voltammetry.  相似文献   

6.
Oxidative stress, caused by reactive oxygen species (ROS), is responsible for modulating several pathological conditions and aging. Soluble and bound phenolic extracts of commonly consumed millets, namely, kodo, finger (Ravi), finger (local), foxtail, proso, little, and pearl, were investigated for their phenolic content and inhibition of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical and ROS, namely, hydroxyl radical, peroxyl radical, hydrogen peroxide (H(2)O(2)), hypochlorous acid (HOCl), and singlet oxygen ((1)O(2)). Inhibition of DPPH and hydroxyl radicals was detrmined using electron paramagnetic resonance (EPR) spectroscopy. The peroxyl radical inhibitory activity was measured using the oxygen radical absorbance capacity (ORAC) assay. The scavenging of H(2)O(2), HOCl, and (1)O(2) was evaluated using colorimetric methods. The results were expressed as micromoles of ferulic acid equivalents (FAE) per gram of grain on a dry weight basis. In addition, major hydroxycinnamic acids were identified and quantified using high-performance liquid chromatography (HPLC) and HPLC-mass spectrometry (MS). All millet varieties displayed effective radical and ROS inhibition activities, which generally positively correlated with phenolic contents, except for hydroxyl radical. HPLC analysis revealed the presence of ferulic and p-coumaric acids as major hydroxycinnamic acids in phenolic extract and responsible for the observed effects. Bound extracts of millet contributed 38-99% to ROS scavenging, depending on the variety and the test system employed. Hence, bound phenolics must be included in the evaluation of the antioxidant activity of millets and other cereals.  相似文献   

7.
Response surface methodology (RSM) was used to predict the optimum conditions of extraction of barley samples (organic solvent percent in the extraction medium, temperature, and time). Antioxidant capacity in the barley meals was highest under optimum extraction conditions of 80.2% methanol and 60.5 degrees C for 38.36 min as predicted by RSM. Phenolic antioxidative compounds of six barley cultivars, namely, Falcon, AC Metcalfe, Tercel, Tyto, Phoenix, and Peregrine, were extracted under the conditions obtained by RSM after defatting with hexane, and subsequently the extracts were assessed for their antioxidant and antiradical activities and metal chelation efficacy. The potential of barley extracts in inhibiting peroxyl and hydroxyl radical induced supercoiled DNA double-strand scission was also studied. Total phenolic content as measured according to Folin-Ciocalteu's method ranged from 13.58 to 22.93 mg of ferulic acid equiv/g of defatted material, with the highest content in Peregrine. Total antioxidant activity as measured by Trolox equivalent antioxidant capacity ranged from 3.74 to 6.82 micromol/g of defatted material. Metal chelation capacity of the extracts as measured by 2,2'-bipyridyl competition assay varied from 1.1 to 2.1 micromol of ethylenediaminetetraacetic acid equiv/g of defatted material. IC(50) values for 1,1-diphenyl-2-picrylhydrazyl radical as measured by electron paramagnetic resonance ranged from 1.51 to 3.33 mg/mL, whereas the corresponding values for hydroxyl radical ranged between 2.20 and 9.65 mg/mL. Inhibition of peroxyl radical induced supercoiled DNA scission ranged from 78.2 to 92.1% at the concentration of 4 mg/mL of extracts, whereas the corresponding values for hydroxyl radical induced DNA scission ranged from 53.1 to 65.3%.  相似文献   

8.
The chemistry behind antioxidant capacity assays   总被引：3，自引：0，他引：3  
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.  相似文献   

9.
Assays comprising three probes for different mechanisms of antioxidant activity in food products have been modified to allow better comparison of the contributions of the different mechanisms to antioxidant capacity (AOC). Incorporation of a common format for oxygen radical absorbance capacity (ORAC), ferric reducing antioxidant power (FRAP), and iron(II) chelating activity (ICA) assays using 96-well microplates provides a comprehensive and high-throughput assessment of the antioxidant capacity of food extracts. The methods have been optimized for aqueous extracts and validated in terms of limit of quantification (LoQ), linearity, and precision (repeatability and intermediate reproducibility). In addition, FRAP and ORAC assays have been validated to assess AOC for lipophilic extracts. The relative standard deviation of repeatability of the methods ranges from 1.2 to 6.9%, which is generally considered to be acceptable for analytical measurement of AOC by in vitro methods. Radical scavenging capacity, reducing capacity, and iron chelating properties of olive mill wastewaters (OMWW), oregano, and parsley were assessed using the validated methods. OMWW showed the highest radical scavenging and reducing capacities, determined by ORAC and FRAP assays, respectively, followed by oregano and parsley. The ability to chelate Fe (2+) was, in decreasing order of activity ( p > 0.05) parsley congruent with oregano > OMWW. Total phenol content, determined by the Folin-Ciocalteu method, correlated to the radical scavenging and reducing capacities of the samples but not to their chelating properties. Results showed that the optimized high-throughput methods provided a comprehensive and precise determination of the AOC of lipophilic and hydrophilic food extracts in vitro.  相似文献   

10.
Phenolic compounds from soft and hard wheat and their milling fractions were extracted into distilled deionized water, and their in vitro antioxidant activities were evaluated. Wheat samples were used as such (nontreated) or subjected to pH adjustment (treated) in order to simulate gastrointestinal pH conditions. The total phenolic content (TPC) was determined using Folin-Ciocalteu's procedure. The total antioxidant activity (TAA) was determined using Trolox equivalent antioxidant capacity assay and expressed as Trolox equivalents. The antioxidant activity of wheat extracts was also evaluated using the beta-carotene bleaching assay, scavenging of 2,2-diphenyl-1-picrylhydrazyl radical, and inhibition of oxidation of human low density lipoprotein cholesterol. The TPC, TAA, and antioxidant potential, evaluated using different methods of wheat samples, were significantly increased following gastrointestinal tract-simulated pH changes. Thus, digestion taking place in the gastrointestinal tract in vivo may also enhance the antioxidant properties of the extracts.  相似文献   

11.
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.  相似文献   

12.
An electron spin resonance (ESR) spectrometry study was conducted to examine the free radical scavenging properties of bran extracts of Alliance and Wichita wheat using hydroxyl radical (HO*), 2,2-diphenyl-1-picryhydrazyl radical (DPPH*), and superoxide radical anion (O2*-) and their chelating capacities against Cu2+. Also reported is the radical cation 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS*+) scavenging activity, oxygen radical absorbing capacity (ORAC), and chelating property against Fe2+ of the bran extracts measured by the spectrophotometric methods. Significant radical scavenging and chelating capacities were detected in the bran extracts, along with significant levels of phenolic acids, tocopherols, and carotenoids. Ferulic acid, with a concentration range of 130.60-146.38 microg/g, was the predominant phenolic acid in all of the tested bran samples and accounted for approximately 53-67% of total phenolic acids on a weight basis. Total tocopherol concentration ranged from 1.87 to 2.95 micromol/100 g of bran, whereas total carotenoid level was 0.20-0.33 micromol/100 g of bran. In addition, both wheat variety and growing conditions might significantly alter antioxidant properties and concentrations of beneficial components in wheat bran.  相似文献   

13.
为明确广东石豆兰的抗氧化能力及抗氧化成分,以鳞茎为材料,以甲醇、乙醇、丙酮和乙酸乙酯为提取剂得到4种提取物,测定了4种提取物中总黄酮、总酚含量及其抗氧化活性,并分析了两者之间的关系。结果表明,石豆兰总酚含量高于总黄酮,乙醇为总酚和总黄酮2种物质的最佳提取溶剂;甲醇提取物对羟基自由基清除力和对Fe2+的螯合力最强,对DPPH自由基清除效果最差;丙酮提取物具有最强的总还原力和清除超氧阴离子的能力,而清除羟基自由基能力最弱;乙酸乙酯提取物清除DPPH自由基的能力最强,但总还原力、清除超氧阴离子的能力、Fe2+的螯合力最弱。此外,除了DPPH自由基外,其他4个抗氧化指标与总黄酮或总酚含量呈正相关(0.185<r<0.969),总还原力与总黄酮含量呈显著相关。本研究结果为广东石豆兰抗氧化物质的提取及进一步开发利用提供了参考依据。  相似文献   

14.
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.  相似文献   

15.
The antioxidant capacity of methanol and ethanol seed extracts from Euterpe oleracea Mart. (a?aí) against the reactive oxygen species (ROS) peroxyl radicals, peroxynitrite, and hydroxyl radicals was studied with the total oxidant scavenging capacity (TOSC) assay in a modified and automated version. Cold methanol digestion was the most efficient extraction method with respect to the antioxidant capacity. The extracts exhibit good antioxidant capacity against peroxyl radicals, similar to the capacity of the pulp. The antioxidant capacity against peroxynitrite and hydroxyl radicals is even higher. The main antioxidants identified by HPLC-MS and HPLC-CEAD are five different procyanidins (di- through pentamers); furthermore, protocatechuic acid and epicatechin were identified as minor compounds. Determination of TOSC values of HPLC seed extract fractions indicates that the procyanidins contribute substantially to the overall antioxidant capacity. In addition, however, other compounds that have not yet been identified are responsible for a large part of the observed antioxidant capacity.  相似文献   

16.
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.  相似文献   

17.
A spectrophotometric method was developed to measure antioxidant free-radical scavenging capacity. Rhodamine B (RhB) was oxidized by hydroxyl radical generated via the Fenton reaction to yield a photoinactive RhB product. RhB absorption at 550 nm was restored when antioxidant agents scavenged hydroxyl radical to protect RhB from oxidation. On the basis of the dose response of antioxidant recovery capacity, a model was developed to calculate the free-radical scavenging ability. This method was sensitive to a wide range of antioxidant activity with ascorbic acid reference set as one; the antioxidant recovery capacity of quercetin was 635 compared to 2 for benzoic acid.  相似文献   

18.
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.  相似文献   

19.
Vitamin C equivalent antioxidant capacity (VCEAC) of phenolic phytochemicals   总被引：11，自引：0，他引：11  
To express the antioxidant capacity of plant foods in a more familiar and easily understood manner (equivalent to vitamin C mg/100 g), two stable radical species, ABTS(*)(-) and DPPH(*), commonly used for antioxidant activity measurements, were employed independently to evaluate their efficacies using apple polyphenolic extracts and seven polyphenolic standards including synthetic Trolox. Their antioxidant activities were expressed as vitamin C equivalent antioxidant capacity (VCEAC) in mg/100 g apple or mg/100 mL of the reference chemical compounds in 10 and 30 min using the ABTS(*)(-) and DPPH(*) scavenging assays, respectively. The antioxidant capacity of Gala apples and seven phenolic standards, determined by both ABTS(*)(-) and DPPH(*) scavenging assays, showed a dose-response of the first-order. Fresh Gala apples had a VCEAC of 205.4 +/- 5.6 mg/100 g using the ABTS assay, and the relative VCEACs of phenolic standards were as follows: gallic acid > quercetin > epicatechin > catechin > vitamin C > rutin > chlorogenic acid > Trolox. With the DPPH radical assay, the VCEAC of fresh Gala apples was 136.0 +/- 6.6 mg/100 g, and the relative VCEACs of seven phenolic standards were, in decreasing order, as follows: gallic acid > quercetin > epicatechin > catechin > or = vitamin C > Trolox > rutin > chlorogenic acid. Because the ABTS assay can be used in both organic and aqueous solvent systems, employs a specific absorbance at a wavelength remote from the visible region, and requires a short reaction time, it is a more desirable method than the DPPH assay. Therefore, it is recommended that antioxidant capacity be expressed as vitamin C mg/100 g equivalent (VCEAC) using the ABTS assay.  相似文献   

20.
Conjugated linoleic acids (CLA) have shown diverse biological activities, including anti-carcinogenic, anti-atherosclerotic, anti-adipogenic, and anti-diabetogenic effects. Recent reports also showed that CLA has free radical scavenging capacity, which may give health benefits to human beings. However, the application of CLA as a bioactive ingredient has been limited due to its insolubility in water. To overcome this problem, we investigated antioxidant activities of arginine (Arg)-CLA, a water-soluble CLA salt, using both 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assays. CLA, Arg, and Arg-CLA all exerted radical scavenging activities in a dose-dependent manner in both assays. Arg-CLA at 20 mM scavenged 89 and 55% of ABTS and DPPH radicals in 3 h, respectively, whereas CLA alone quenched only 48 and 26% of them under the same conditions. The antioxidant activity of the Arg-CLA complex was found to be synergistic in ABTS assay and comparable to that of vitamin E in DPPH assay.  相似文献   

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