Cellular antioxidant activity of common fruits

Measurement of antioxidant activity using biologically relevant assays is important in the screening of fruits for potential health benefits. The cellular antioxidant activity (CAA) assay quantifies antioxidant activity in cell culture and was developed to meet the need for a more biologically representative method than the popular chemistry antioxidant capacity measures. The objective of the study was to determine the cellular antioxidant activity, total phenolic contents, and oxygen radical absorbance capacity (ORAC) values of 25 fruits commonly consumed in the United States. Pomegranate and berries (wild blueberry, blackberry, raspberry, and blueberry) had the highest CAA values, whereas banana and melons had the lowest. Apples were found to be the largest contributors of fruit phenolics to the American diet, and apple and strawberries were the biggest suppliers of cellular antioxidant activity. Increasing fruit consumption is a logical strategy to increase antioxidant intake and decrease oxidative stress and may lead to reduced risk of cancer.     

Controlled-atmosphere effects on postharvest quality and antioxidant activity of cranberry fruits

The effects of controlled-atmosphere (CA) storage on the firmness, respiration rate, quality, weight loss, total phenolics and flavonoids contents, and total antioxidant activities of the Pilgrim and Stevens cultivars of cranberries (Vaccinium macrocarpon Aiton) have been studied during storage in atmospheres of 2, 21, and 70% O(2) with 0, 15, and 30% CO(2) (balance N(2)); and 100% N(2) at 3 degrees C. Elevated CO(2) concentrations decreased bruising, physiological breakdown, and decay of berries, thereby reducing fruit losses. Respiration and weight loss of fruits decreased, but fruit softening increased, at higher CO(2) concentrations. Accumulations of acetaldehyde, ethanol, and ethyl acetate varied by cultivar and storage atmosphere but were generally highest in the 2 and 70% O(2) and 100% N(2) atmospheres and increased in response to elevated CO(2) concentrations. Overall, the 30% CO(2) plus 21% O(2) atmosphere appeared optimal for the storage of cranberries. Sensory analysis is required, however, to confirm that accumulations of fermentation products at this atmosphere are acceptable for consumers. Stevens fruits had a higher phenolics content and total antioxidant activity than Pilgrim fruits. The storage atmosphere did not affect the content of total phenolics or flavonoids. However, the total antioxidant activity of the fruits increased overall by about 45% in fruits stored in air. This increase was prevented by storage in 30% CO(2) plus 21% O(2).     

Flavanol content and antioxidant activity in winery byproducts

Proanthocyanidins, particularly those coming from wine and grape products, have became of interest to nutritionists. Particular attention is currently being paid to the exploitation of this kind of grape byproducts for obtaining bio-active phenolic compounds with potential application as food antioxidants and preventive agents against cancer and other diseases. In this work, the flavanol composition of various winery byproducts submitted to different degrees of industrial exploitation has been studied and their antioxidant activity determined using two different methods (TBARS and TEAC) to evaluate their interest as suitable sources for the preparation of flavanol-rich antioxidant extracts. All the byproducts studied were still good flavanol sources no matter their exploitation degree. An important conclusion was that dried grape seeds, obtained as an end byproduct after the color extraction and alcohol distillation of the wine pomace, still kept important flavanol concentrations and significant antioxidant activity, even if they were submitted to high temperatures. These byproducts can be considered a cheap source for the extraction of antioxidant flavanols, which can be used as dietary supplements or in the production of phytochemicals.     

Phenolic constituents in the fruits of Cinnamomum zeylanicum and their antioxidant activity

Defatted cinnamon fruit powder was successively extracted with benzene ethyl acetate, acetone, MeOH, and water. The concentrated water extract contained the maximum amount of phenolics and showed the highest antioxidant activities. Hence, it was fractionated by Diaion HP-20SS, Diaion HP-20, and Sephadex LH-20 column chromatographies. It gave five purified compounds, the purities of which were analyzed by HPLC. Compounds 1-5 were identified as 3,4-dihydroxybenzoic acid (protocatechuic acid), epicatechin-(2beta-->O-7,4beta-->8)-epicatechin-(4beta-->8)-epicatechin (cinnamtannin B-1), 4-[2,3-dihydro-3-(hydroxymethyl)-5-(3-hydroxypropyl)-7-(methoxy)benzofuranyl]-2-methoxyphenyl beta-d-glucopyranoside (urolignoside), quercetin-3-O-(6-O-alpha-l-rhamnopyranosyl)-beta-d-glucopyranoside (rutin), and quercetin-3-O-alpha-l-rhamnopyranoside by using extensive spectral studies. The antioxidant activities of purified compounds were screened for their antioxidative potential using beta-carotene-linoleate and 1,1-diphenyl-2-picrylhydrazyl model systems. All of the compounds showed antioxidant and radical scavenging activities. This is the first report of the isolation and identification of nonvolatile constituents and as well as antioxidant activities from cinnamon fruits.     

Spectroscopic identification and antioxidant activity of glucosylated carotenoid metabolites from Cydonia vulgaris fruits

Carotenoid metabolites are common plant constituents with significant importance for the flavor and aroma of fruits. Three new carotenoid derivatives, (2E,4E)-8-hydroxy-2,7-dimethyl-2,4-decadiene-1,10-dioic acid 1-O-beta-D-glucopyranosyl ester (1), (2Z,4E)-8-beta-D-glucopyranosyloxy-2,7-dimethyl-2,4-decadiene-1,10-dioic acid (3), and 3,9-dihydroxymegastigmast-5-ene-3-O-[beta-D-glucopyranosyl-(1-->6)]-beta-D-glucopyranoside (5), as well as three known compounds, have been isolated from the ethanolic extract of peels of Cydonia vulgaris, the fruit of a shrub belonging to the same family as the apple. All the compounds were identified by spectroscopic techniques, especially 1D and 2D NMR. Antioxidant activities of all the isolated metabolites were assessed by measuring their ability to scavenge DPPH radical and superoxide radical (O2*-) and to induce the reduction of Mo(VI).     

Changes in antioxidant content of tomato fruits in response to cultivar and nutrient solution composition

The aim of this study was to investigate the effect of cationic proportions (K/Ca/Mg) in the nutrient solution on fruit quality (quality attributes and antioxidant content) using a high-pigment, 'Lunarossa', tomato cultivar and a standard tomato cultivar ('Corfù') grown in soilless culture. Treatments were defined by a factorial combination of three nutrient solutions having different cationic proportions and two indeterminately growing round tomato cultivars. A high proportion of K in the nutrient solution increased the quality attributes (fruit dry matter, total soluble solids content) and the lycopene content of tomato fruit, whereas a high proportion of Ca improved tomato fruit yield and reduced the incidence of blossom-end rot (BER). The highest total antioxidant activity was observed in the treatment with a high proportion of Mg in the Lunarossa cultivar. The high-pigment hybrid has provided a higher antioxidant content (lycopene and alpha-tocopherol content) than the commercial hybrid, but it was more susceptible to BER and consequently less productive.     

Processing effects on lycopene content and antioxidant activity of tomatoes

Consumption of tomato products has been associated with decreased risk of some cancer types, and the tomato antioxidant, lycopene, is thought to play an important role in the observed health effects. In this study, four carotenoids, trans-lycopene, phytofluene, phytoene, and zeta-carotene, were quantified in tomato products. Samples of raw tomatoes, tomato juice after hot break scalder, and final paste were obtained from two different processing plants over two years. Comparison of carotenoid levels throughout processing indicated that lycopene losses during processing of tomatoes into final paste (25-30 degrees Brix) ranged from 9 to 28%. The initial Brix level of the raw tomatoes appeared to influence the amount of lycopene loss that occurred, possibly due to the differences in processing time required to achieve the final desired Brix level of the paste. In general, no consistent changes in the other carotenoids were observed as a function of processing. The antioxidant activity of fresh tomatoes, tomato paste, and three fractions obtained from these products (i.e., aqueous, methanol, and hexane fractions) was also determined. In both a free radical quenching assay and a singlet oxygen quenching assay, significant antioxidant activity was found in both the hexane fraction (containing lycopene) and the methanol fraction, which contained the phenolic antioxidants caffeic and chlorogenic acid. The results suggest that in addition to lycopene, polyphenols in tomatoes may also be important in conferring protective antioxidative effects.     

Distribution of conjugated and free phenols in fruits: antioxidant activity and cultivar variations

Total and free phenolic contents of 16 commonly consumed fruits (comprising 9 apples, 4 pears, and one each of peach, plum, and kiwi fruit cultivars) were measured by Folin-Ciocalteu assay. Total phenol contents varied from 272 to 475 mg of CtE/100 g of fresh weight. Of the apple cultivars studied, Braeburn and Empire had the highest and lowest total phenol content, respectively. The apple cultivars ranked in the following decreasing order: Braeburn > Red Delicious > Crisp Pink > Granny Smith > Royal Gala > Bramley > Golden Delicious > Fuji > Empire. Among pear cultivars, the order was Forelle > Taylor's > Peckham's > Conference. Peach and plum equally had high contents. The percentage of conjugated phenolics ranged between 3 (Red Delicious) and 77% (Empire) of the total phenols. Comparison of different cultivars of the same fruit and between different fruits showed broad variations in both phenolic content and in vitro antioxidant activity; a weak correlation (R(2) = 0.58) was observed between the phenolic content of the fruits and the total antioxidant activity, as estimated by the FRAP assay. The calculated dietary intake of total, free, and conjugated phenols from average per capita consumption of apples and pears in different regions of the U.K. varied between 104-126, 53-64, and 51-62 mg of CtE/day, respectively.     

Determination of the total antioxidant activity of fruits and vegetables by a liposome assay

The effects of mixtures of antioxidants on the oxidation of phospholipids have been investigated in large unilamellar liposomes following initiation by 2,2'-azobis(2-aminopropane) dihydrochloride. The lag phase increased linearly with antioxidant concentration. The lag phases of mixtures containing alpha-tocopherol with ascorbic acid showed synergy between the antioxidants, but mixtures of beta-carotene with alpha-tocopherol or ascorbic acid were not synergistic. The liposome system was used to investigate the total antioxidant activity of lipid- and water-soluble extracts from 16 samples of fruits, vegetables, and related food products. The water-soluble extracts caused greater increases in lag phase than the lipid-soluble extracts. The lag phase of liposomes containing the water-soluble extracts from fruits and vegetables increased linearly with the total phenolic concentration, with the continental salad extract having the longest lag phase. The lipid-soluble extract from apples caused the largest increase in lag phase of the lipid-soluble extracts. The lag phases of the lipid-soluble and water-soluble extracts of all fruits and vegetables studied were additive, but no synergy was detected. The lag phase of the liposomes containing both the water-soluble and lipid-soluble extracts varied from 611.5 min for the continental salad extracts to 47.5 min for the cauliflower extracts.     

Sources of antioxidant activity in Australian native fruits. Identification and quantification of anthocyanins

Selected native Australian fruits, muntries (Kunzea pomifera F. Muell., Myrtaceae), Tasmanian pepper berry (Tasmanian lanceolata R. Br., Winteraceae), Illawarra plum (Podocarpus elatus R. Br. ex Endl., Podocarpaceae), Burdekin plum (Pleiogynium timorense DC. Leenh, Anacardiaceae), Cedar Bay cherry (Eugenia carissoides F. Muell., Myrtaceae), Davidson's plum (Davidsonia pruriens F. Muell. var. pruriens, Davidsoniaceae), and Molucca raspberry (Rubus moluccanus var. austropacificus van Royen, Rosaceae), were evaluated as sources of antioxidants by 2,2-diphenyl-1-picrylhydrazyl and ferric reducing antioxidant power assays and compared with blueberry (Vaccinum spp. cv. Biloxi). The total reducing capacity of five fruits was 3.5-5.4-fold higher than that of blueberry, and the radical scavenging activities of muntries and Burdekin plum were 1.5- and 2.6-fold higher, respectively. The total phenolic level by Folin-Ciocalteu assay highly correlated with the antioxidant activity. Therefore, systematic research was undertaken to identify and characterize phenolic complexes. In the present study we report on the levels and composition of anthocyanins. The HPLC-DAD and HPLC/ESI-MS-MS (ESI = electrospray ionization) analyses revealed simple anthocyanin profiles of one to four individual pigments, with cyanidin as the dominating type. This is the first evaluation of selected native Australian fruits aiming at their utilization for the development of novel functional food products.     

Squalene content and antioxidant activity of Terminalia catappa leaves and seeds

Squalene was identified by gas chromatography-mass spectrometry and high-performance liquid chromatography (HPLC) spiking analyses in the supercritical CO(2) extracts of freeze-dried abscisic leaves of Terminalia catappa L. When the freeze-dried abscisic, senescent, mature, and immature leaves and seeds were subjected to supercritical CO(2) extraction at 40 degrees C and 3000 psi and HPLC quantitation, squalene contents were 12.29, 2.42, 1.75, 0.9, and 0% in the extracts and corresponding to 1499, 451, 210, 65, and 0 microg/g in the freeze-dried sample, respectively. When the extracts were applied for antioxidative characterization by supplementation in an iron/ascorbate system with linoleic acid and in a pork fat storage system for inhibition of conjugated diene hydroperoxide (CDHP) formation or in a free radical scavenging system with 1,1-diphenyl-2-picryl-hydrazyl (DPPH), the extracts of leaves exhibited potent antioxidative and DPPH scavenging activities and increased with an increase of leaf maturity. However, the seed extracts only exhibited potent inhibition of CDHP formation and very low DPPH scavenging activity.     

Enzyme-catalyzed change of antioxidants content and antioxidant activity of asparagus juice

A pectolytic enzyme preparation from Aspergillus niger (pectinase AN) decreased most rutin content and antioxidant activity of asparagus juice. To investigate the mechanism of such loss, we analyzed several possible related enzyme activities in pectinase AN. We found that the activity of pectinase AN to oxidize guaiacol had no significant difference with or without the presence of H2O2; thus it was laccase activity, not peroxidase (PO) activity, that pectinase AN contained. We did not find any polyphenol oxidase (PPO) activity in pectinase AN. Laccase in pectinase AN could be the major cause of loss of rutin and antioxidant activity of asparagus juice. When most laccase activity of pectinase AN was inactivated after heating at 70 degrees C for 1.5 min and incubated with asparagus juice, the loss rate of rutin was only 9% of that treated with unheated pectinase AN, and the antioxidant activity was even increased. Rhamnosidase activity was detected in pectinase AN and can change rutin in asparagus juice to quercetin-3-glucoside, which has higher antioxidant activity than rutin. This may explain the increase of antioxidant activity of asparagus juice treated with heated pectinase AN that still contained some rhamnosidase activity. The discovery of our research is helpful to produce juice with high antioxidant activity and high health benefits in the juice industry.     

Total phenolic content, antioxidant activity, and cross-cultural consumer rejection threshold in white and red wines functionally enhanced with catechin-rich extracts

White and red wines spiked with catechin-rich green tea extract and grape seed extract were assessed for phenolic content, antioxidant activity, and cross-cultural consumer rejection thresholds in relation to wine as a functional food. Health functionality is an important factor in functional foods, and spiking pure compounds or plant extracts is an effective method to increase or control functionality. The total phenolic content and antioxidant activity were measured in wines spiked to different extract concentrations, namely, control and 50, 100, 200, 400, and 800 mg/L, to confirm the dose-response curves in both white and red wines. Consumer rejection thresholds (CRTs) were established for spiked wines in a Korean and in an Australian population. Our results showed that the green tea extract and grape seed extract increased the antioxidant activity dose dependently, and the CRTs varied considerably between the Korean and the Australian groups, with Koreans preferring wines spiked with green tea extract and Australians showing a preference for wines spiked with grape seed extract. These results have implications for producing wine products that are enhanced in phenolic compounds and targeted to different cultural groups.     

Effect of Aspergillus oryzae-challenged germination on soybean isoflavone content and antioxidant activity

Application of microbial stress to soybean during germination induces the accumulation of phytoalexins, which have many health benefits. In this study, the effects of stress induced by Aspergillus oryzae on the phytochemical composition of germinating soybeans were investigated, and their radical scavenging activity was compared with those of ungerminated (US) and germinated (GS) soybeans. Additionally, the antioxidant activity of coumestrol, a soybean phytoalexin, against hydrogen peroxide-induced reactive oxygen species (ROS) was investigated in HepG2 cells. A. oryzae exposure significantly decreased the total isoflavone content and induced coumestrol and glyceollin I. A. oryzae-challenged germinated soybeans exhibited the highest radical scavenging activity (IC(50) = 0.55 mg/mL) as compared to US and GS. Coumestrol exhibited significantly higher radical scavenging activity than daidzein and genistein. Furthermore, coumestrol significantly prevented hydrogen peroxide-induced ROS production and lipid peroxidation and inhibited decreases in cell viability, intracellular glutathione (GSH) levels, and superoxide dismutase (SOD) activity. These results indicate that using food-grade A. oryzae to elicit the biosynthesis of phytoalexins alters the secondary metabolite profiles of the soybeans and offers enhanced bioactivity of soybean as a functional food ingredient.     

臭氧精准处理提高采后草莓抗氧化酶活性和酚类物质含量

臭氧处理被认为是一种有效而无污染的贮藏保鲜方式,而臭氧的强氧化性也被认为对于采后果蔬的活性氧清除系统存在影响。该研究以天津市种植最广泛的"京桃香"草莓为试材,探讨了不同浓度的臭氧(0、2.144、6.432、10.72、15.008 mg/m~3)处理对草莓采后活性氧起抑制作用的非酶促、酶促清除系统的影响。总酚(total phenols,TPC)、总类黄酮(total flavonoids,TFC)和抗坏血酸(ascorbic acid,ASA)为非酶促抗氧化关键代谢物指标,以超氧化物歧化酶(superoxide dismutase,SOD)、过氧化物酶(peroxidase,POD)和抗坏血酸超氧化物酶(ascorbic acid superoxidase,APX)为关键酶活指标进行检测。结果表明,整个贮藏期间,对于非酶促清除系统,采后草莓总酚类物质在2.144、6.432、15.008 mg/m~3的臭氧处理组具有一定的累积效果,但低于对照组的含量;黄酮类物质在2.144、15.008 mg/m~3处理组中的含量不断下降,而6.432mg/m~3处理组中的含量为先下降后上升,但3个处理组中的含量均低于对照组;ASA含量在对照组中的变化不大,但在2.144、6.432、15.008 mg/m~3处理组中不断减少,且15.008 mg/m~3的臭氧处理组的下降幅度最大。而相较于对照组和其他3个不同浓度的臭氧处理组,10.72 mg/m~3的O_3处理能够显著促进酚类化合物和抗坏血酸的生成,维持总黄酮含量在较高水平。对于酶促清除系统,高浓度的O_3处理能够快速诱导草莓果实POD活性的升高。但是随着贮藏时间的延长,15.008mg/m~3处理组迅速出现降低的变化趋势。相较于对照组,2.144 mg/m~3浓度O_3处理组的酶活变化不大,但6.432 mg/m~3处理组却存在一定的抑制作用。10.72 mg/m~3的O_3处理能够显著提升SOD活性(P﹤0.05),使其最大值提前,能够提升APX活性,刺激POD酶活的活性提高,并推迟POD活性出现高峰的时间。总体来说,10.72 mg/m~3浓度的臭氧处理能够增强采后草莓氧化还原系统相关酶促反应酶活,促进非酶促反应抗氧化物质的生成,对于采后草莓的抗氧化物质积累具有积极作用,有利于提高草莓的食用价值和经济价值。     

Cyclooxygenase inhibitory and antioxidant compounds from crabapple fruits

Crabapple trees belong to the Malus genus (Rosaceae) and bear fruits that are sparingly consumed and used in the preparation of fruit beverages. Cyclooxygenase (COX) enzyme inhibitory and antioxidant bioassay-guided fractionation of the aqueous and methanol extracts of Malus x kornicensis and Malus x Indian Summer yielded (+)-catechin (1), (-)-epicatechin (2), cyanidin-3-O-beta-galactopyranoside (3), and amygdalin (4). Pure compounds 1-4 were obtained by HPLC, identified by LC-ES/MS, CD, and NMR spectroscopic methods and evaluated for their COX enzyme inhibitory and antioxidant activities. In COX-1 and -2 enzyme inhibitory assays, compounds 1-3 (all at 80 microM) showed activities of 20.4, 46.3%; 57.6, 47.9%; and 8.2, 13.7%, respectively, compared to naproxen (54.3, 41.3%; 10 microM), ibuprofen (47.5, 39.8%; 10 microM), Celebrex (46.2, 66.3%; 1.67 ppm), and Vioxx (23.8, 88.1%, 1.67 ppm). In the antioxidant assay, the catechins (1-2) and anthocyanin (3) (all at 40 microM) showed activities of 61.3, 62.5, and 60.1%, respectively. The synthetic antioxidants, tert-butylhydroquinone (TBHQ), butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), and vitamin E (all tested at 10 microM), gave 75.2, 80.1, 70.0, and 10.2% activities, respectively. The cyanogenic glycoside, amygdalin (4), and its hydrolysis products, mandelonitrile (5) and benzaldehyde (6), were not active in the antioxidant or COX enzyme inhibitory assays at 80 microM concentrations.     

Determination of antioxidant activity of wine byproducts and its correlation with polyphenolic content

It has been demonstrated that wine and other products derived from the grape have a high antioxidant capability; as a possible consequence of this, they may have potential benefits for health. The byproducts of the winemaking process represent a source of antioxidant compounds that has been relatively unexploited to date, but that is now the subject of increasing industrial interest. This article describes an approach to the study of the antioxidant activity of grape marcs, stalks, and dregs of both white and red varieties. This activity is compared with the measurements of their content of total polyphenols and of individual polyphenolic compounds, identified and quantified by HPLC. From the results we have been able to establish a positive correlation between the antioxidant activity and the total polyphenolic content of samples, but not with specific compounds.     

Effects of processing steps on the phenolic content and antioxidant activity of beer

A new analytical method (liquid chromatography-antioxidant, LC-AOx) was used that is intended to separate beer polyphenols and to determine the potential antioxidant activity of these constituents after they were allowed to react online with a buffered solution of the radical cation 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS(?+)). Using the LC-AOx method, it was possible to demonstrate that the extent of the antioxidant activity was very much dependent on the phenolic compound considered. The method was also applied to the analysis of beer extracts and allowed the evaluation of their antioxidant activity at different steps of beer processing: brewing, boiling, and fermentation. This study showed that the total antioxidant activity remained unchanged throughout beer processing, as opposed to the polyphenolic content, which showed a 3-fold increase. Hopping and fermentation steps were the main causes of this increase. However, the increase measured after fermentation was attributed to a better extraction of polyphenols due to the presence of ethanol, rather than to a real increase in their content. Moreover, this method allowed the detection of three unknown antioxidant compounds, which accounted for 64 ± 4% of the total antioxidant activity of beer and were individually more efficient than caffeic acid and epicatechin.