Unravelling the total antioxidant capacity of pinotage wines: contribution of phenolic compounds

The total antioxidant capacity (TAC) and phenolic composition of 139 Pinotage wines (2002 and 2003 vintages) were determined using the 2,2'-azino-di(3-ethylbenzo-thialozine-sulfonic acid) scavenging assay and high-performance liquid chromatography, respectively. The contribution of individually quantified phenolic compounds to the wine TAC was calculated using their concentrations and Trolox equivalent antioxidant capacity (TEAC) values. The TEAC values of quercetin-3-galactoside, isorhamnetin, and peonidin-3-glucoside are reported for the first time. Between 11 and 24% of the measured TAC of Pinotage wines was explained by the sum of the calculated contributions of their quantified phenolic compounds comprising monomeric phenolic compounds and procyanidin B1. Ultrafiltration was carried out to attempt separation of monomeric and polymeric phenolic compounds. Analysis of ultrafiltration permeates and retentates enabled estimation of the TAC contribution of large molecular weight (MW) unknown compounds (46%) (>50 kDa), including oligomeric and polymeric phenolic compounds and small MW unknown compounds (34%) (<50 kDa). Three mixtures, containing 12 phenolic compounds in typical concentrations expected in Pinotage wines, exhibited 16-23% synergistic antioxidant activity. This suggests that synergy between phenolic compounds does play a role in the wine TAC but that it does not explain the large discrepancy between measured and calculated TAC values.     

Unfermented rooibos tea: quantitative characterization of flavonoids by HPLC-UV and determination of the total antioxidant activity

Unfermented rooibos originates from the leaves and the stems of the indigenous South African plant, Aspalathus linearis, and it has been reported to have a higher content of flavonoids compared to that of fermented rooibos. The HPLC/UV method developed in our laboratory for the analysis of the fermented rooibos was applied to the quantitative characterization of the major flavonoids present in the unfermented rooibos. Main compounds determined were aspalathin (49.92 +/- 0.80 mg/g), isoorientin (3.57 +/- 0.18 mg/g), orientin (2.336 +/- 0. 049 mg/g), and rutin (1.69 +/- 0.14 mg/g), followed in order by isovitexin, vitexin, isoquercitrin and hyperoside, quercetin, luteolin and chrysoeryol. The identity of detected flavonoids was confirmed by comparing their retention times and UV spectra with those of corresponding standards. The total antioxidant activity (TAA) of the tea infusions was measured by the ABTS*+ radical cation decolorization assay. The TAA of unfermented rooibos (0.8 Trolox meq/g) resulted 2-fold higher than that of the fermented rooibos. When compared with different water infusions of Camellia sinensis (green and black tea), this TAA value was about 50% lower.     

Major phenolics in apple and their contribution to the total antioxidant capacity

The contribution of each phytochemical to the total antioxidant capacity of apples was determined. Major phenolic phytochemicals of six apple cultivars were identified and quantified, and their contributions to total antioxidant activity of apples were determined using a 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging assay and expressed as vitamin C equivalent antioxidant capacity (VCEAC). Average concentrations of major phenolics and vitamin C in six apple cultivars were as follows (mg/100 g of fresh weight of apples): quercetin glycosides, 13.20; procyanidin B(2), 9.35; chlorogenic acid, 9.02; epicatechin, 8.65; phloretin glycosides, 5.59; vitamin C, 12.80. A highly linear relationship (r (2) > 0.97) was attained between concentrations and total antioxidant capacity of phenolics and vitamin C. Relative VCEAC values of these compounds were in the order quercetin (3.06) > epicatechin (2.67) > procyanidin B(2) (2.36) > phloretin (1.63) > vitamin C (1.00) > chlorogenic acid (0.97). Therefore, the estimated contribution of major phenolics and vitamin C to the total antioxidant capacity of 100 g of fresh apples is as follows: quercetin (40.39 VCEAC) > epicatechin (23.10) > procyanidin B(2) (22.07) > vitamin C (12.80) > phloretin (9.11) > chlorogenic acid (8.75). These results indicate that flavonoids such as quercetin, epicatechin, and procyanidin B(2) rather than vitamin C contribute significantly to the total antioxidant activity of apples.     

Effect of freezing and storage on the phenolics,ellagitannins, flavonoids,and antioxidant capacity of red raspberries

Scottish-grown red raspberries are a rich source of vitamin C and phenolics, most notably, the anthocyanins cyanidin-3-sophoroside, cyanidin-3-(2(G)-glucosylrutinoside), and cyanidin-3-glucoside, and two ellagitannins, sanguiin H-6 and lambertianin C, which are present together with trace levels of flavonols, ellagic acid, and hydroxycinnamates. The antioxidant capacity of the fresh fruit and the levels of vitamin C and phenolics were not affected by freezing. When fruit were stored at 4 degrees C for 3 days and then at 18 degrees C for 24 h, mimicking the route fresh fruit takes after harvest to the supermarket and onto the consumer's table, anthocyanin levels were unaffected while vitamin C levels declined and those of elligitannins increased, and overall, there was no effect on the antioxidant capacity of the fruit. It is concluded, therefore, that freshly picked, fresh commercial, and frozen raspberries all contain similar levels of phytochemicals and antioxidants per serving.     

Correlation between some nutritional components and the total antioxidant capacity measured with six different assays in eight horticultural crops

The contents of antioxidant nutritional compounds, total soluble phenolics (TSP), vitamin C, vitamin E, beta-carotene, and total carotenoids (TC), were correlated with the total antioxidant capacity (AOC) of hydrophilic (HPE) and lipophilic extracts (LPE) from eight horticultural crops, namely, guava, avocado, black sapote, mango, papaya, prickly pear fruit, cladodes, and strawberry. AOC was measured using six different assays: 2,2'-diphenyl-1-picrylhydrazyl (DPPH), N,N-dimethyl-p-phenylendiamine (DMPD), ferric-ion-reducing antioxidant power (FRAP), oxygen radical absorbance capacity (ORAC), Trolox equivalent antioxidant capacity (TEAC), and total oxidant scavenging capacity (TOSC). AOC values from HPE were about 95 times higher than LPE values. HPE of guava had the highest AOC value when evaluated with DMPD, DPPH, FRAP, TEAC, and TOSC assays, whereas with ORAC assay, black sapote had the highest value. HPE of papaya and prickly pear fruit presented the lowest AOC values with all assays. From HPE, vitamin C and TSP contents were highly correlated with AOC for all assays, while from LPE, TC and beta-carotene contents possessed a high correlation with AOC only in the DMPD assay.     

Contribution of individual polyphenolics to total antioxidant capacity of plums

To study the effect of polyphenolics on antioxidant capacities of plums, the amounts of total phenolics, total flavonoids and individual phenolic compounds, and vitamin C equivalent antioxidant capacity (VCEAC) of eleven plum cultivars was determined. There was a good linear relationship between the amount of total phenolics and total antioxidant capacity (r2 = 0.9887). The amount of total flavonoids and total antioxidant capacity also showed a good correlation (r2 = 0.9653). Although the summation of individual antioxidant capacity was lower than the total antioxidant capacity of plum samples, there was a positive correlation (r2 = 0.9299) of total antioxidant capacity of plum samples with the sum of the VCEACs calculated from individual phenolics. Chlorogenic acids and glycosides of cyanidin, peonidin, and quercetin were major phenolics among eleven plum cultivars. The antioxidant capacity of chlorogenic acids and anthocyanins showed higher correlation (r2) of 0.7751 and 0.6616 to total VCEAC, respectively, than that of quercetin glycosides (r2 = 0.0279). Chlorogenic acids were a major source of antioxidant activity in plums, and the consumption of one serving (100 g) of plums can provide antioxidants equivalent to 144.4-889.6 mg of vitamin C.     

Nitrogen effects on total flavonoids,chlorogenic acid,and antioxidant activity of the medicinal plant Chrysanthemum morifolium

Chrysanthemum morifolium (Ramat.) has a long history of cultivation and use as a traditional medicine and tea plant in China. A greenhouse experiment with potted soil–quarz mixture studied the effects of nitrogen supply (0, 56, 112, 167, 224, 334, 501, 556, and 668 mg N kg^–1) on concentrations and ratios of total flavonoids and chlorogenic acid in the flowers of C. morifolium using spectrophotometric and HPLC methods. The antioxidant activity of the flowers was determined as the radical scavenging activities of hydroxyl, superoxide anion, and 2,2‐diphenyl‐1‐picrylhydrazyl hydrate (DPPH) free radicals. A high N supply decreased the concentrations of total flavonoids by 18%–35% and that of chlorogenic acid by 8%–60% compared to a low N‐supply rate. At the same time, increasing N supply significantly decreased the antioxidant activity of the flowers. The antioxidant activity of C. morifolium flowers was significantly positively correlated with the concentrations of total flavonoids and chlorogenic acid. We conclude that an N supply in excess of 300 mg (kg soil)^–1 will negatively affect the antioxidant activity and thereby reduce the quality of C. morifolium flowers.     

Major flavonoids in grape seeds and skins: antioxidant capacity of catechin, epicatechin, and gallic acid

Grape seeds and skins are good sources of phytochemicals such as gallic acid, catechin, and epicatechin and are suitable raw materials for the production of antioxidative dietary supplements. The differences in levels of the major monomeric flavanols and phenolic acids in seeds and skins from grapes of Vitis vinifera varieties Merlot and Chardonnay and in seeds from grapes of Vitis rotundifolia variety Muscadine were determined, and the antioxidant activities of these components were assessed. The contribution of the major monomeric flavonols and phenolic acid to the total antioxidant capacity of grape seeds and skins was also determined. Gallic acid, monomeric catechin, and epicatechin concentrations were 99, 12, and 96 mg/100 g of dry matter (dm) in Muscadine seeds, 15, 358, and 421 mg/100 g of dm in Chardonnay seeds, and 10, 127, and 115 mg/100 g of dm in Merlot seeds, respectively. Concentrations of these three compounds were lower in winery byproduct grape skins than in seeds. These three major phenolic constituents of grape seeds contributed <26% to the antioxidant capacity measured as ORAC on the basis of the corrected concentrations of gallic acid, catechin, and epicatechin in grape byproducts. Peroxyl radical scavenging activities of phenolics present in grape seeds or skins in decreasing order were resveratrol > catechin > epicatechin = gallocatechin > gallic acid = ellagic acid. The results indicated that dimeric, trimeric, oligomeric, or polymeric procyanidins account for most of the superior antioxidant capacity of grape seeds.     

Correlation of selected constituents with the total antioxidant capacity of coffee beverages: influence of the brewing procedure

Relationships between volatile and nonvolatile compounds and the antioxidant capacity of coffee brews prepared from commercial conventional and torrefacto roasted coffees, employing commonly used doses and prepared by four brewing procedures (filter, plunger, mocha, and espresso machine) were assessed. Significant correlations between volatile Maillard reaction products and antioxidant capacity (measured by both 2,2-diphenyl-1-picrylhydrazyl radical and redox potential methods) were not observed. Highly positive correlations between browned compounds and caffeine with both antioxidant capacity parameters were reported. Principal component analysis allowed coffee brews separation according to coffee roasting processes (PC1) and brewing procedures (PC2), showing that in all cases coffee brews from torrefacto roasted coffee were more antioxidant that those extracted from conventional ones; also, coffee brews extracted by an espresso machine were more antioxidant than those extracted by mocha, plunger, and filter machines.     

Ellagitannins,flavonoids, and other phenolics in red raspberries and their contribution to antioxidant capacity and vasorelaxation properties

Analysis of extracts of Glen Ample raspberries (Rubus idaeus L.) by gradient, reverse phase HPLC with diode array and tandem mass spectrometry identified eleven anthocyanins, including cyanidin-3-sophoroside, cyanidin-3-(2(G)-glucosylrutinoside), cyanidin-3-glucoside, cyanidin-3-rutinoside, pelargonidin-3-sophoroside, pelargonidin-3-(2(G)-glucosylrutinoside), and pelargonidin-3-glucoside. Significant quantities of an ellagitannin, sanguiin H-6, with an M(r) of 1870 were detected along with lower levels of a second ellagitannin, lambertianin C, which has an M(r) of 2804. Other phenolic compounds that were detected included trace levels of ellagic acid and its sugar conjugates along with one kaempferol- and four quercetin-based flavonol conjugates. Fractionation by preparative HPLC revealed that sanguiin H-6 was a major contributor to the antioxidant capacity of raspberries together with vitamin C and the anthocyanins. Vasodilation activity was restricted to fractions containing lambertianin C and sanguiin H-6.     

Phenotypic divergence in vegetable amaranth for total antioxidant capacity,antioxidant profile,dietary fiber,nutritional and agronomic traits

A lot of variations in vegetable amaranth germplasm have been observed in Bangladesh. It has been used as a cheap source of antioxidants, nutrients, protein, and dietary fiber. But no efforts had not been taken to know the status of antioxidant content, polyphenol, flavonoid, antioxidant vitamins and minerals, dietary fiber, nutritional and agronomic traits. In this study, Forty-three vegetable amaranth genotypes were evaluated to determine the status of total antioxidant content, polyphenol, flavonoid, antioxidant vitamins and minerals, dietary fiber, nutritional and agronomic traits and the magnitude of genetic diversity based on the contribution of those traits for meaningful grouping and proper utilization in future breeding program. The experiment was carried out in an open experimental field at Bangabandhu Sheikh Mujibur Rahman Agricultural University, Bangladesh in a randomized complete block design with three replications. Multivariate (Principal component and cluster) analysis was done using numerical taxonomic techniques of Sneath, & Sokal. Four principal components contributed 98.61% of the variation. Biological yield and total antioxidant content was strongly associated with their related all agronomic traits. Total flavonoid content had a higher contribution to total antioxidant capacity compared to vitamin and mineral antioxidants. Contribution of antioxidant profile and agronomic traits was the highest in diversity of vegetable amaranth. Both high and low yielding genotypes had a high antioxidant profile. Therefore, high yielding genotypes (From cluster VI) could be used directly as high antioxidant profile varieties and low yielding genotypes as a source of donor parents in hybridization program. Cluster analysis grouped the genotypes into six clusters. The diverse genotypes in different clusters were identified. Genotypes with desirable genes of one cluster hybridized with promising genotypes of other diverge clusters could facilitate the accumulation of favorable genes in hybrids.

Abbreviations: PCA: principal component analysis; PC: principal component; dw: dry weight; TFC: total flavonoid content; TAC: total antioxidant capacity; TPC: total polyphenol content; K: potassium; Ca: calcium; Mg: magnesium; Fe: iron; Mn: manganese; Cu: copper; Zn: zinc     

Processing strawberries to different products alters contents of vitamin C, total phenolics, total anthocyanins, and antioxidant capacity

Strawberries were processed to juice, nectar, wine, and puree. For investigation of the antioxidant capacity as well as the contents of ascorbic acid, total phenolics and total anthocyanins, samples were taken after different stages of production to determine the effects of processing. The content of vitamin C was measured spectrophotometrically. The total phenolic content was analyzed by using the Folin-Ciocalteu method, and the amount of total anthocyanins was determined by using the pH-differential method. Two different methods-the trolox equivalent antioxidant capacity assay and the ferric reducing antioxidant power test-were used to determine the hydrophilic antioxidant capacity. This study showed the decrease of all investigated parameters within processing strawberries to different products. The content of ascorbic acid decreased with production time and processing steps, especially during heat treatment. The investigations on total phenolics in strawberry products proved fining to be a mild method to clarify berry juices and wines without removing high amounts of total phenolics. Fermentation did not lead to heavy losses of total phenolics, probably due to polymerization and condensation of monomer phenolics such as anthocyanins. Total anthocyanins and the hydrophilic antioxidant capacity decreased while using high temperatures. Anthocyanins also decreased considerably during the processing of wines, mainly caused by fermentation and pasteurization.     

HPLC method for the quantification of procyanidins in cocoa and chocolate samples and correlation to total antioxidant capacity.

Monomeric and oligomeric procyanidins present in cocoa liquors and chocolates were separated and quantified in four different laboratories using a normal-phase high-performance liquid chromatography (HPLC) method with fluorescence detection. Procyanidin standards through decamers were obtained by extraction from cocoa beans, enrichment by Sephadex LH-20 gel permeation chromatography, and final purification by preparative normal-phase HPLC. The purity of each oligomeric fraction was assessed using HPLC coupled to mass spectrometry. A composite standard was then prepared, and calibration curves were generated for each oligomeric class using a quadratic fit of area sum versus concentration. Results obtained by each of the laboratories were in close agreement, which suggests this method is reliable and reproducible for quantification of procyanidins. Furthermore, the procyanidin content of the samples was correlated to the antioxidant capacity measured using the ORAC assay as an indicator for potential biological activity.     

New investigation of the isothermal oxidation of extra virgin olive oil: determination of free radicals, total polyphenols, total antioxidant capacity, and kinetic data

As a follow-up of the research programs carried out by our group concerning the artificial isothermal rancidification process in extra virgin olive oil (EVOO), in the present work the trends of both the total antioxidant capacity and the total polyphenols concentration as well as the main kinetic parameters of the process during the thermal oxidation of EVOO were studied and compared. In addition, the possibility of evaluating the increase in radicals concentration during the thermal oxidation process using a superoxide dismutase biosensor was also studied. The present investigation concerning this important food product is highly topical as it refers to the state of alteration of the EVOO used for cooking or frying, as a function of the temperature reached.     

马兰黄酮类化合物的提取及其抗氧化活性

为了探索以亚临界水萃取马兰黄酮类化合物的效果,以得率为指标,通过单因素和正交试验,优选其萃取工艺参数,并对所得黄酮类化合物的抗氧化性能进行测定。结果表明,在压力为5.0 MPa,温度160℃、提取时间25 min、水料比为20∶1 mL/g的条件下马兰黄酮类化合物的得率为9.01%。与水和乙醇回流提取法（马兰黄酮类化合物得率分别为5.12%和7.13%）相比,亚临界水萃取在提取时间和得率方面均有明显的优势,所得马兰黄酮类化合物在一定浓度范围内具有较强的清除自由基和抗氧化能力。研究结果为马兰黄酮类化合物的开发提供参考。     

Interactions between fertilizer nitrogen and soil nitrogen—the so-called 'priming' effect

Experiments with ¹⁵N labelled fertilizers often show that plants given fertilizer N take up more N from the soil than plants not given N—the priming effect or ‘added nitrogen interaction’(ANI). This paper is a theoretical study of ANIs and how they can affect the interpretation of experiments with ¹⁵N labelled fertilizers. ANIs can be ‘Real’, if for example, fertilizer N increases the volume of soil explored by roots, or ‘apparent’, caused by pool substitution or by isotope displacement reactions. Pool substitution is the process by which added labelled N stands proxy for native unlabelled N that would otherwise have been removed from that pool. Microbial immobilization of N, whether driven by the decomposition of soil organic matter or by the decomposition of plant roots, can lead to pool substitution and is the dominant cause of apparent ANIs. Denitrification and plant uptake of N can also, under special circumstances, lead to pool substitution and thus give rise to apparent ANIs. Isotope displacement reactions, in which the added labelled N displaces native unlabelled N from a ‘bound’ pool, can lead to apparent ANIs but are only likely to be of significance in exceptional circumstances. The relationship between ANIs, ‘A’ values and N fertilizer uptake efficiencies are examined by means of a simple model for uptake of ¹⁵N-labelled fertilizer by a crop. A positive ‘apparent’ ANI is accompanied by an ‘A’ value that changes as fertilizer applications increase. Likewise, a positive ‘apparent’ ANI also causes fertilizer uptake efficiency to appear lower when measured by the uptake of ¹⁵N than when measured by the non-isotopic ‘difference’ method.     

Effect of processing on the antioxidant vitamins and antioxidant capacity of Vigna sinensis Var. Carilla

Cowpea (Vigna sinensis L. var. Carilla) flours obtained by fermentation with inoculum Lactobacillus plantarum (PF) or with the natural microorganisms present in the flour (NF) and subsequent heat treatment in an autoclave were prepared to study the effect of fermentation on the antioxidant vitamin content and on the antioxidant capacity. Bacterial counts and pH values, vitamins C and E, carotenoids, glutathione (GSH), superoxide dismutase-like activity (SOD-like activity), peroxyl radical-trapping capacity (PRTC), lipid peroxidation in unilamillar liposomes, and Trolox equivalent antioxidant capacity (TEAC) were evaluated in raw and processed cowpea flours. gamma-Tocopherol and delta-tocopherol were found in raw cowpea, whereas vitamin C and carotenoids were not detected. An increase in the vitamin E activity was observed in PF, whereas vitamin C and carotenoids were not detected in fermented cowpea flours. Fermentation or heat treatment in an autoclave after fermentation produced processed cowpea flours with lower PRTC, glutathione content, and SOD-like activity than those of the raw seeds. However, those processes increased the capacity to inhibit the lipid peroxidation in unilamellar lipoposomes and TEAC. According to the results obtained in this study, the fermentation of cowpeas (naturally or with L. plantarum) and fermentation and subsequent heat treatment in an autoclave are good processes to obtain functional cowpea flours having higher antioxidant capacity than the raw legume.     

Structure-activity relationships of flavonoids in the cellular antioxidant activity assay

Chemical antioxidant activity assays are used extensively to evaluate the potential bioactivity of plant foods and their phytochemical constituents, but they do not mimic the complexity of biological systems. The cellular antioxidant activity (CAA) activity assay was developed to be a more biologically relevant model to measure antioxidant activity. Structure-activity relationships of flavonoids have been determined in many chemistry antioxidant activity assays, and they vary with the protocols. The objective of this study was to determine structure-activity relationships of selected flavonoids in the CAA assay. The structures that conferred flavonoids with the most antioxidant activity in the CAA assay were a 3',4'- o-dihydroxyl group in the B-ring, a 2,3-double bond combined with a 4-keto group in the C-ring, and a 3-hydroxyl group. Isoflavones had no cellular antioxidant activity. Flavanols with a galloyl moiety had higher antioxidant activity than those without, and a B-ring 3',4',5'-trihydroxyl group further improved their efficacy. ORAC values for flavonoids were not related to their CAA values. Knowledge of structure-activity relationships in the CAA assay may be helpful in assessing potential in vivo antioxidant activity of flavonoids.     

Antioxidant capacity and phenolic content of grapes, sun-dried raisins, and golden raisins and their effect on ex vivo serum antioxidant capacity

Grapes and raisins provide phenolic antioxidants, which contribute to their potential health benefits. The objectives of this study were to compare the antioxidant capacity and phenolic content of green Thompson seedless grapes (the most common variety of grapes consumed in the United States), sun-dried raisins, and golden raisins (both produced from Thompson seedless grapes) and to observe the effects of their consumption over 4 weeks in 15 healthy human males with a cross-over design. The oxygen radical absorbance capacity (ORAC) (positive statistical significance for grapes after 2 weeks and golden raisins after 3 weeks), serum oxidation (positive statistical significance for golden raisin lag time after 4 weeks), total phenolics (no significant effects), and C-reactive protein (no significant effects) were monitored. Immediately postconsumption, there were some significant nonpositive changes. It is hypothesized that these negative results may be explained by postprandial oxidation, a known effect after carbohydrate consumption. Golden raisins had the highest antioxidant capacity and phenolic content. The consumption of a serving of grapes or raisins each day, in addition to a typical diet, may not be sufficient to overcome postprandial oxidation when consumed with other high carbohydrate foods but may have beneficial antioxidant effects over time.     

Electrochemical sensing of total antioxidant capacity and polyphenol content in wine samples using amperometry online-coupled with microdialysis

This work describes the method for total antioxidant capacity (TAC) and/or total content of phenolics (TCP) analysis in wines using microdialysis online-coupled with amperometric detection using a carbon microfiber working electrode. The system was tested on 10 selected wine samples, and the results were compared with total reactive antioxidant potential (TRAP), oxygen radical absorbance capacity (ORAC), and chemiluminescent determination of total antioxidant capacity (CL-TAC) methods using Trolox and catechin as standards. Microdialysis online-coupled with amperometric detection gives similar results to the widely used cyclic voltammetry methodology and closely correlates with ORAC and TRAP. The problem of electrode fouling is overcome by the introduction of an electrochemical cleaning step (1-2 min at the potential of 0 V vs Ag/AgCl). Such a procedure is sufficient to fully regenerate the electrode response for both red and white wine samples as well as catechin/Trolox standards. The appropriate size of microdialysis probes enables easy automation of the electrochemical TAC/TCP measurement using 96-well microtitration plates.