Assessment of radical scavenging capacity and lipid peroxidation inhibiting capacity of antioxidant

The role of radical scavenging antioxidants against oxidative stress has received much attention, and the antioxidant capacity has been assessed by various methods. Among them, a method that measures the effect of antioxidant on decay of the probe is one of the most widely used methods. The present study was performed to compare the two methods to assess the antioxidant capacity, one to follow the decay of the probe and the other to measure lipid peroxidation products in human plasma. It was shown that the method following probe decay was suitable for assessment of radical scavenging capacity of antioxidant, but not for the capacity to inhibit lipid peroxidation in plasma. This is true whether a hydrophilic or lipophilic probe is used. Such different results arise from the fact that the efficacy of inhibition of lipid peroxidation by antioxidants depends on the fate of antioxidant-derived radical and interaction between antioxidants as well as the capacity of free radical scavenging. Thus, the capacity of antioxidants for inhibition of lipid peroxidation should be assessed from the effect on the extent of oxidation, not from the effect on probe decay.     

Stoichiometric and kinetic studies of phenolic antioxidants from Andean purple corn and red-fleshed sweetpotato

Stoichiometric and kinetic values of phenolics against DPPH (2,2-diphenyl-1-picrylhydrazyl) were determined for Andean purple corn (Zea mays L.) and red sweetpotato (Ipomoea batatas). Both crops had higher antioxidant capacity and antiradical kinetics than blueberries and higher or similar anthocyanin and phenolic contents. The second-order rate constant (k(2)) was 1.56, 1.12, 0.57, and 0.26 (mg antiradical/mL)(-1) s(-1) for red sweetpotato, Trolox, purple corn, and blueberry, respectively. On the molar basis of active hydroxyl groups, k(2)' showed the same order as for k(2). Corn cob and sweetpotato endodermis contributed the most in phenolic compounds and antioxidant capacity. Both crops studied can be considered as excellent novel sources of natural antioxidants for the functional food and dietary supplement markets.     

Quince (Cydonia oblonga Miller) fruit (pulp, peel, and seed) and Jam: antioxidant activity

To study the antioxidant activity of quince fruit (pulp, peel, and seed) and jam, methanolic extracts were prepared. Each extract was fractionated into a phenolic fraction and an organic acid fraction and was analyzed by high-performance liquid chromatography (HPLC)/diode array detection and HPLC/UV, respectively. Antiradical activities of the extracts and fractions were evaluated by a microassay using 1,1'-diphenyl-2-picrylhydrazyl. The phenolic fraction always exhibited a stronger antioxidant activity than the whole methanolic extract. Organic acid extracts were always the weakest in terms of antiradical activity, which seems to indicate that the phenolic fraction gives a higher contribution for the antioxidant potential of quince fruit and jam. The evaluation of the antioxidant activity of methanolic extracts showed that peel extract was the one presenting the highest antioxidant capacity. The IC50 values of quince pulp, peel, and jam extracts were correlated with the caffeoylquinic acids total content. Among the phenolic fractions, the seed extract was the one that exhibited the strongest antioxidant activity. The IC50 values of quince pulp, peel, and jam phenolic extracts were strongly correlated with caffeoylquinic acids and phenolics total contents. For organic acid fractions, the peel extract was the one that had the strongest antiradical activity. The IC50 values of quince pulp, peel, and jam organic acid fractions were correlated with the ascorbic acid and citric acid contents.     

Interaction among phenols in food fortification: negative synergism on antioxidant capacity

In this work, a study about the consequences of the interaction among phenol compounds on antioxidant capacity is proposed. The antiradical activity evolution of an ethanol solution containing a mixture of three monophenols (catechin, resveratrol, and quercetin) was compared with the trend followed by each single phenol at three different temperatures (22, 37, and 60 degrees C). An initial increase and a following decrease in antioxidant activity were observed for all solutions at the three temperatures. The lower antiradical activity values of the mixture in comparison to the controls during the entire period of storage clearly showed that interaction among these polyphenols promotes a negative synergistic effect on this property.     

Bioguided isolation and identification of the nonvolatile antioxidant compounds from fennel (Foeniculum vulgare Mill.) waste

A bioguided isolation of an aqueous extract of fennel waste led to the isolation of 12 major phenolic compounds. Liquid chromatography coupled to atmospheric pressure chemical ionization mass spectrometry (LC/UV/APCI-MS) combined with spectroscopic methods (NMR) was used for compound identification. Radical scavenging activity was tested using three methods: DPPH*, superoxide nitro-blue tetrazolium hypoxanthine/xanthine oxidase, and *OH/luminol chemiluminescence. In addition to products described in the literature, eight antioxidant compounds were isolated and identified for the first time in fennel: 3-caffeoylquinic acid, 4-caffeoylquinic acid, 1,5-O-dicaffeoylquinic acid, rosmarinic acid, eriodictyol-7-O-rutinoside, quercetin-3-O-galactoside, kaempferol-3-O-rutinoside, and kaempferol-3-O-glucoside. The structures of eriodictyol-7-O-rutinoside and quercetin-3-O-glucuronide were completely elucidated by two-dimensional NMR experiments. The isolated compounds exhibited a strong antiradical scavenging activity, which may contribute to the interpretation of the pharmacological effects of fennel.     

New method to determine antioxidant activity of polyphenols

The capacity of polyphenolic compounds to reduce the beta-carotene-linoleic acid cooxidation enzymatically induced by soybean lipoxygenase was assayed to determine their comprehensive antioxidant ability. The inhibition of the coupled oxidation is a well-known spectrophotometric method for the antioxidant activity measurement. A modification of this method is proposed to reduce assay time and to gain simplicity. The antioxidant abilities of several polyphenols were determined, and quercetin and sinapic acid were most active. These results were compared to those obtained by the DPPH* procedure to evaluate the free radical scavenging contribution to the total protective action. The highest values of antiradical activity were found for ellagic and quercetin.     

Optimization of the extraction of antioxidative constituents of six barley cultivars and their antioxidant properties

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%.     

Commercial dietary antioxidant supplements assayed for their antioxidant activity by different methodologies

Seven commercial dietary antioxidant supplements were evaluated for their in vitro antioxidant capacity by different methodologies: antiradical activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH.), inhibition of methyl linoleate (MeLo) autoxidation, and resistance to ion-dependent oxidation of human low-density lipoprotein (LDL). Great variability in antioxidant activity was seen among the supplements, as well as different patterns of antioxidant capacity depending on the method used. The same orders of activity were found for the MeLo autoxidation and LDL-induced oxidation methods, but the differences recorded by the former were wider. "Food/compound intake equivalents" of the doses recommended by the manufacturers were calculated in terms of quercetin, strawberries, and red wine for each of the supplements studied. Food/compound intake equivalents varied between 7.9 and 190 mg of quercetin according to the DPPH. scavenging method, between 0.20 and 98 mg according to the MeLo autoxidation method, and between 3.4 and 83 mg according to LDL-induced oxidation. In equivalent terms of red wine, the food/compound intake equivalents varied between 7 and 159 mL, between 3 and 1354 mL, and between 4 and 89 mL for the same three methods. In terms of equivalents of strawberry, they varied between 14 and 343 mg according to the DPPH. scavenging method and between 57 g and 26 kg according to the MeLo autoxidation method. These results show the need to standardize dietary supplements in terms of their antioxidant capacity to match required doses to the oxidative status of consumers.     

Patatin,the tuber storage protein of potato (Solanum tuberosum L.), exhibits antioxidant activity in vitro

The potato (Solanum tuberosum L.) tuber storage protein, patatin, was purified to homogeneity with a molecular mass of 45 kDa. The purified patatin showed antioxidant or antiradical activity by a series of in vitro tests, including 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical (half-inhibition concentration, IC(50), was 0.582 mg/mL) scavenging activity assays, anti-human low-density lipoprotein peroxidation tests, and protections against hydroxyl radical-mediated DNA damages and peroxynitrite-mediated dihydrorhodamine 123 oxidations. Using electron paramagnetic resonance (EPR) spectrometry for hydroxyl radical detections, it was found that the intensities of the EPR signal were decreased by the increased amounts of patatin added (IC(50) was 0.775 mg/mL). Through modifications of patatin by iodoacetamide or N-bromosuccinimide, it was found that the antiradical activities of modified patatin against DPPH or hydroxyl radicals were decreased. It was suggested that cysteine and tryptophan residues in patatin might contribute to its antioxidant activities against radicals.     

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.     

Variation in phenolic composition and antioxidant activity during flower development of safflower (Carthamus tinctorius L.)

This work was aimed to study the effect of extraction solvent system with varying polarities on polyphenol, flavonoid and proanthocyanidin contents and DPPH scavenging activity. Obtained results showed that phenolic contents and antioxidant activities varied considerably as function of solvent polarity. The extraction with acetone/water (2:8) showed the highest flower polyphenol content (15.09 mg GAE/g DW). Moreover, antiradical capacities against DPPH, chelating power and lipid peroxidation assay were maximal in acetone/water (2:8) of flower extract. Significant variation in antioxidant properties was observed between different development stages of Carthamus tinctorius flowers; the highest antioxidant activity was observed at stage III (full flowering) while phenolic composition reached its maximum at stage II (flower formation). Gallic acid was the most abundant phenolic compound in C. tinctorius orange flowers, accounting for about 102.57 (μg/g DW). Findings underline the potential health benefits as a result of consuming C. tinctorius flowers and suggest that it could be used as valuable flavor with functional properties for food or nutraceutical products on the basis of the high polyphenol contents and antioxidant activities.     

Effect of principal polyphenolic components in relation to antioxidant characteristics of aged red wines.

Aged red wines possess significantly different polyphenolic composition compared with young ones, mainly due not only to formation of polymeric compounds but also because of oxidation, hydrolysis, and other transformations that may occur in native grape phenolics during aging. Representative Greek, single-variety, aged red wines were examined for total phenol, total flavanol, and total anthocyanin content using spectrophotometry, and attempts were made to establish correlations with the antiradical, reducing, and hydroxyl free radical scavenging activity. In addition, HPLC analyses were carried out, to ascertain whether individual polyphenols are actually responsible for the antioxidant effects of aged red wines. It was found that total flavanols are the class of polyphenols that account for hydroxyl free radical scavenging efficacy and to a lesser extent for antiradical and reducing ability, whereas there was a less significant link between the antioxidant properties and the total phenolics and only a weak relationship to total anthocyanin content. The correlation of the antioxidant properties with the principal polyphenols showed that individual compounds are weakly associated with all the antioxidant parameters, suggesting that the expression of antioxidant activity in aged red wines is rather a consequence of synergism between various phenolics, and it is not simply attributed to specific constituents.     

Melanoidins exert a weak antiradical activity in watery fluids

The Maillard reaction has a dramatic impact on the overall acceptance and nutritional quality of most of the foods consumed daily in European countries. Melanoidins are polymeric structures formed in the last stage of the Maillard reaction with nearly unknown effect on the human health. The antiradical activity of several melanoidins isolated from model systems and coffee has been studied. A novel antiradical efficiency concept has been applied to describe the antiradical activity in an aqueous medium by bleaching the radical cation N,N-dimethyl-p-phenylenediamine (DMPD(*)(+)). Melanoidins exerted a significantly lower antiradical activity than classical antioxidant compounds (tannic acid, ferrulic acid, caffeic acid, gallic acid, and Trolox) in an aqueous medium. Significant differences have been observed according to the type of amino acid used as reactant during the formation of the melanoidin structure and the antiradical efficiency exerted.     

Isolation of an in vitro and ex vivo antiradical melanoidin from roasted barley

The antiradical properties of water-soluble components of both natural and roasted barley were determined in vitro, by means of DPPH* assay and the linoleic acid-beta-carotene system, and ex vivo, in rat liver hepatocyte microsomes against lipid peroxidation induced by CCl4. The results show the occurrence in natural barley of weak antioxidant components. These are able to react against low reactive peroxyl radicals, but offer little protection against stable DPPH radicals deriving from peroxidation in microsomal lipids. Conversely, roasted barley yielded strong antioxidant components that are able to efficiently scavenge free radicals in any system used. The results show that the barley grain roasting process induces the formation of soluble Maillard reaction products with powerful antiradical activity. From roasted barley solution (barley coffee) was isolated a brown high molecular mass melanoidinic component, resistant to acidic hydrolysis, that is responsible for most of the barley coffee antioxidant activity in the biosystem.     

Total antioxidant capacity of teas by the ferric reducing/antioxidant power assay

This study aimed to compare in vitro antioxidant power of different types of tea (Camellia sinensis). The ferric reducing/antioxidant power (FRAP) assay was used to measure the total antioxidant power of freshly prepared infusions of 25 types of teas. Results showed that different teas had widely different in vitro antioxidant power and that the antioxidant capacity was strongly correlated (r = 0. 956) with the total phenolics content of the tea. Expressed as micromol of antioxidant power/g of dried tea leaves, values ranged as 132-654 micromol/g for black ("fermented") teas, 233-532 micromol/g for Oolong ("semifermented") teas, and 272-1144 micromol/g for green ("nonfermented") teas. One cup of tea of usual strength (1-2%), therefore, can provide the same potential for improving antioxidant status as around 150 mg of pure ascorbic acid (vitamin C).     

Antioxidative and antiproliferative properties of selected barley (Hordeum vulgarae L.) cultivars and their potential for inhibition of low-density lipoprotein (LDL) cholesterol oxidation

Aqueous methanolic extracts of whole kernels from six different barley cultivars, namely, Falcon, AC Metcalfe, Tyto, Tercel, Phoenix, and Peregrine, were examined for their total phenolic content (TPC), oxygen radical scavenging capacity (ORACFL), hydroxyl radical scavenging capacity (HORACFL), potency in prevention of lipid oxidation using the Rancimat method, efficacy in inhibition of Cu(II)-induced human LDL cholesterol oxidation, and antiproliferative activities using Caco-2 colorectal adenocarcinoma cell line. Total phenolic content as measured by Folin-Ciocalteu's method ranged from 0.68 to 1.19 mg of ferulic acid equiv/g of defatted material, whereas ORACFL and HORACFL values were 11.28-19.10 and 9.06-12.99 micromol of Trolox equiv/g of defatted material, respectively. Protection factor (PF), a measure of the effect of extracts on the prevention of oxidation of stripped corn oil as measured by Rancimat, ranged from 0.97 to 1.59. Furthermore, barley extracts showed 19.64-33.93% inhibition against Cu(II)-induced human LDL cholesterol oxidation at a final concentration of 0.02 mg/mL. The proliferation of Caco-2 colon cancer cells was significantly (p < 0.05) inhibited in a dose-dependent fashion in the presence of all barley extracts tested at the end of the day 4 of incubation. At the end of day 4, barley extracts rendered 29.3-51.2 and 9.3-15.9% inhibition of cell proliferation at 0.5 and 0.05 mg/mL, respectively. Phenolic extracts from whole barley kernel tested possessed high antioxidant, antiradical, and antiproliferative potentials. Therefore, inclusion of whole barley into the daily diet may render beneficial health benefits.     

枯草芽孢杆菌Y-6产抗菌肽的体外抗氧化效果研究

为研究枯草芽孢杆菌Y-6产抗菌肽的抗氧化活性,通过化学发光法测定抗菌肽清除H2O2、·OH和O2·3种氧自由基的性能,再通过分光光度法测定抗菌肽的还原能力、对DPPH·的清除率以及对肝匀浆丙二醛(MDA)生成的抑制率.结果表明,抗菌肽有一定清除自由基、抑制脂质过氧化的作用,试验范围内抗菌肽浓度越高,抗氧化效果越好.抗菌...     

Antioxidant activity of the main phenolic compounds isolated from hot pepper fruit (Capsicum annuum L)

Four cultivars (Bronowicka Ostra, Cyklon, Tornado, and Tajfun) of pepper fruit Capsicum annuum L. were studied for phenolics contents and antioxidant activity. Two fractions of phenolics, flavonoids (with phenolic acids) and capsaicinoids, were isolated from the pericarp of pepper fruit at two growth stages (green and red) and were studied for their antioxidant capacity. Both fractions from red fruits had higher activities than those from green fruits. A comparison of the capsaicinoid fraction with the flavonoid and phenolic acid fraction from red fruit with respect to their antioxidant activity gave similar results. Phenolic compounds were separated and quantified by LC and HPLC. Contents of nine compounds were determined in the flavonoid and phenolic acid fraction: trans-p-feruloyl-beta-d-glucopyranoside, trans-p-sinapoyl-beta-d-glucopyranoside, quercetin 3-O-alpha-l-rhamnopyranoside-7-O-beta-d-glucopyranoside, trans-p-ferulyl alcohol-4-O-[6-(2-methyl-3-hydroxypropionyl] glucopyranoside, luteolin 6-C-beta-d-glucopyranoside-8-C-alpha-l-arabinopyranoside, apigenin 6-C-beta-d-glucopyranoside-8-C-alpha-l-arabinopyranoside, lutoeolin 7-O-[2-(beta-d-apiofuranosyl)-beta-d-glucopyranoside], quercetin 3-O-alpha-l-rhamnopyranoside, and luteolin 7-O-[2-(beta-d-apiofuranosyl)-4-(beta-d-glucopyranosyl)-6-malonyl]-beta-d-glucopyranoside. The main compounds of this fraction isolated from red pepper were sinapoyl and feruloyl glycosides, and the main compound from green pepper was quercetin-3-O-l-rhamnoside. Capsaicin and dihydrocapsaicin were the main components of the capsaicinoid fraction. A high correlation was found between the content of these compounds and the antioxidant activity of both fractions. Their antioxidant activities were elucidated by heat-induced oxidation in the beta-carotene-linoleic acid system and the antiradical activity by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) decoloration test. The highest antioxidant activity in the beta-carotene-linoleic acid system was found for trans-p-sinapoyl-beta-d-glucopyranoside, which was lower than the activity of free sinapic acid. Quercetin 3-O-alpha-l-rhamnopyranoside had the highest antiradical activity in the DPPH system, which was comparable to the activity of quercetin. The activities of capsaicin and dihydrocapsaicin were similar to that of trans-p-feruloyl-beta-d-glucopyranoside in the DPPH model system.     

Antioxidant and antiradical activities in extracts of hazelnut kernel (Corylus avellana L.) and hazelnut green leafy cover

Phenolic compounds in the aqueous systems were extracted, from hazelnut kernel (HK) and hazelnut green leafy cover (HGLC), with 80% (v/v) ethanol (HKe and HGLCe) or 80% (v/v) acetone (HKa and HGLCa). The extracts were examined for their phenolic and condensed tannin contents and phenolic acid profiles (free and esterified fractions) as well as antioxidant and antiradical activities by total antioxidant activity (TAA), antioxidant activity in a beta-carotene-linoleate model system, scavenging of DPPH (2,2-diphenyl-1-picrylhydrazyl) radical, and reducing power. Significant differences (p < 0.05) in the contents of total phenolics, condensed tannins, and TAA existed among the extracts that were examined. HGLCa extract had the highest content of total phenolics (201 mg of catechin equivalents/g of extract), condensed tannins (542 mg of catechin equivalents/g of extract), and TAA (1.29 mmol of Trolox equivalents/g of extract) followed by HGLCe, HKa, and HKe extracts, respectively. Five phenolic acids (gallic acid, caffeic acid, p-coumaric acid, ferulic acid, and sinapic acid) were tentatively identified and quantified, among which gallic acid was the most abundant in both free and esterified forms. The order of antioxidant activity in a beta-carotene-linoleate model system, the scavenging effect on DPPH radical, and the reducing power in all extracts were in the following order: HGLCa > HGLCe > HKa > HKe. These results suggest that both 80% ethanol and acetone are capable of extracting phenolics, but 80% acetone was a more effective solvent for the extraction process. HGLC exhibited stronger antioxidant and antiradical activities than HK itself in both extracts and could potentially be considered as an inexpensive source of natural antioxidants.     

Antioxidant activity of ethanolic extracts from several asparagus cultivars

Three different methods (antiradical activity, inhibition of primary oxidation, and ferric reducing power) have been used to evaluate the antioxidant activity of eight different asparagus cultivars and byproducts: white and green asparagus from Alcalá del Río (Guadalquivir Valley, Seville) and American hybrids, native spears, and their byproducts from Huétor-Tájar (Vega de Granada). The correlation between antioxidant activity and total phenol content was studied. Six standards were also tested to validate the modified methods for antioxidant activity determination. Results obtained for antiradical capacity and reducing power were very similar, and a high correlation with phenols was found (R > or = 0.9 for both tests). Sample origin was an important factor, spears from Huétor-Tájar having higher values (ARC between 7 and 10 and P(R) of 0.25-0.33) than those from Alcalá del Río (ARC 0.6-2 and P(R) of 0.05-0.07). Significant differences were found between spears with the same origin, suggesting that genetics are another factor to take into account. Asparagus inhibits lipid primary oxidation, but no correlation between the inhibition percentage and phenols was observed. Asparagus origin was the only factor that led to significant differences: samples from Huétor-Tájar had higher values (POIC between 18 and 32) than those from Alcalá del Río (POIC of 5-9). Byproducts from the canning industry at Huétor-Tájar were also assayed for antioxidant activity; the results obtained suggested that byproducts could be considered as an excellent source of natural antioxidants.