Effects of different cooking methods on nutritional and physicochemical characteristics of selected vegetables

The objective of the present study was to evaluate the effect of three common cooking practices (i.e., boiling, steaming, and frying) on phytochemical contents (i.e., polyphenols, carotenoids, glucosinolates, and ascorbic acid), total antioxidant capacities (TAC), as measured by three different analytical assays [Trolox equivalent antioxidant capacity (TEAC), total radical-trapping antioxidant parameter (TRAP), ferric reducing antioxidant power (FRAP)] and physicochemical parameters of three vegetables (carrots, courgettes, and broccoli). Water-cooking treatments better preserved the antioxidant compounds, particularly carotenoids, in all vegetables analyzed and ascorbic acid in carrots and courgettes. Steamed vegetables maintained a better texture quality than boiled ones, whereas boiled vegetables showed limited discoloration. Fried vegetables showed the lowest degree of softening, even though antioxidant compounds were less retained. An overall increase of TEAC, FRAP, and TRAP values was observed in all cooked vegetables, probably because of matrix softening and increased extractability of compounds, which could be partially converted into more antioxidant chemical species. Our findings defy the notion that processed vegetables offer lower nutritional quality and also suggest that for each vegetable a cooking method would be preferred to preserve the nutritional and physicochemical qualities.     

Determination of total content of phenolic compounds and their antioxidant activity in vegetables--evaluation of spectrophotometric methods

This research studies in detail the contents of phenolic compounds determined by the Folin-Ciocalteu reagent and the antioxidant activities determined by the TEAC (Trolox equivalent antioxidant capacity), DPPH (using diphenyl-p-picrylhydrazyl radical), and FRAP (ferric reducing antioxidant power) methods, and their correlations for used standards with these methods (catechine, gallic acid, caffeic acid, ferulic acid, Trolox, ascorbic acid, and ferrous sulfate) and extracts from several species of commonly consumed vegetables were studied in detail. The comparison of absolute values of absorption coefficients for used standards and for individual methods allows one to choose optimal common standards for methods to be compared. The procedures applied for the same sets of the extracts using identical calibration procedures and common standards allowed better comparison of the results obtained by the TEAC, DPPH, and FRAP methods. The values of content of phenolic substances and total antioxidant activity of the sets of samples correlate very well for all used methods. The very high values of antioxidant activity were found in intensely colored vegetables (red cabbage, red onion, etc.), and the values were very low in watery vegetables such as potato, marrow, and cucumber.     

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.     

Antioxidant capacity of some herbs/spices from cameroon: a comparative study of two methods

This study evaluates the antioxidant capacity of 14 herbs/spices from Cameroon. Freeze-dried samples extracted in methanol (free or unconjugated polyphenol) and in 1.2 M hydrochloric acid (HCl) in methanol (total antioxidant that is both unconjugated and conjugated) were analyzed using two different antioxidant assay methods [Folin-Ciocalteu reagent (Folin) and the ferric reducing antioxidant power (FRAP)]. The 1.2 M HCl in methanol extracts had significantly higher (P < 0.001) antioxidant capacities than the methanolic extract. Generally, the FRAP antioxidant values were significantly (P < 0.001) higher than the Folin antioxidant values. Although a significant correlation (P < 0.05) was obtained between the Folin phenol and the FRAP antioxidant, the trends of the antioxidant capacity of the samples were different for the Folin and FRAP methods. The leaves of the Piper species top the total antioxidant tables in both Folin and FRAP assay methods, respectively. Irvingia gabonensis tops the FRAP free antioxidant list, while Piper umbellatum leads the Folin free antioxidant followed by Thymus vulgaris. Thus, the antioxidant capacity of plant samples determined by different methods should be interpreted with caution. However, irrespective of the assay method used, the samples were rich in antioxidants.     

Evaluation of radical scavenging activity of fresh and air-dried tomatoes by three model reactions.

The radical scavenging activity and the antioxidant content of fresh and air-dried tomatoes were investigated. Tomato halves were dried in a pilot-scale dryer under the following conditions: air temperature, 80 degrees C; air flow rate, 1.5 m/s; drying time, 400 min; final moisture, 25%. Carotenoid (lycopene, beta-carotene, lutein) and ascorbic acid were analyzed by HPLC with a spectrophotometric and an electrochemical detector, respectively. Total phenolics were determined by using the Folin-Ciocalteu reagent. The radical scavenging activity was studied in three model systems: (a) the xanthine oxidase and xanthine system, which generates superoxide radical and hydrogen peroxide; (b) the 3-morpholinosydnonimine system, which releases spontaneously superoxide radical and nitrogen monoxide, forming peroxynitrite; (c) the linoleic acid and CuSO(4) system, which promotes lipid peroxidation. These model systems allow the simulation of key reactions involved in the pathogenesis of certain chronic diseases and may be related to the in vivo activity of tomato antioxidants. Hence, these measurements can be used for optimizing tomato processing and storage. The drying process resulted in a decrease of ascorbic acid content, whereas phenol reagent reducing compounds increased. Carotenoid levels were substantially unchanged upon drying. Fresh and air-dried tomato extracts could act as radical scavengers both in the reactive oxygen species-mediated reactions and in lipid peroxidation. Drying affected the antioxidant effectiveness as measured in the xanthine/xanthine oxidase system, which was found to be the most sensitive method for the measurement of tomato antioxidant activity (lower I(50)) but retained the antioxidant effectiveness in the other two systems.     

Antioxidant activity of dietary polyphenols as determined by a modified ferric reducing/antioxidant power assay

Most nonenzymatic antioxidant activity (scavenging of free radicals, inhibition of lipid peroxidation, etc.) is mediated by redox reactions. The antioxidant (AO) activity of polyphenols (PPs), as ferric-reducing power, was determined for the first time using a modified FRAP (ferric reducing/antioxidant power) assay. Reaction was followed for 30 min, and both Fe(II) standards and samples were dissolved in the same solvent to allow comparison. Selected representative PPs included flavonoids (quercetin, rutin, and catechin), resveratrol, tannic acid, and phenolic acids (gallic, caffeic, and ferulic). Carotenoids (beta-carotene and zeaxanthine), ascorbic acid, Trolox, and BHA were included for comparison. Equivalent concentration 1 (EC(1)), as the concentration of AO with a reducing effect equivalent to 1 mmol/L Fe(II), was used to compare AO efficiency. PPs had lower EC(1) values, and therefore higher reducing power, than ascorbic acid and Trolox. Tannic acid and quercetin had the highest AO capacity followed by gallic and caffeic acids. Resveratrol showed the lowest reducing effect. Carotenoids had no ferric reducing ability. Polyphenol's AO efficiency seemed to depend on the extent of hydroxylation and conjugation.     

Modified 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (abts) method to measure antioxidant capacity of Selected small fruits and comparison to ferric reducing antioxidant power (FRAP) and 2,2'-diphenyl-1-picrylhydrazyl (DPPH) methods

The measurement of antioxidant capacity in fruits differs from that of other biological samples due to their low pH and very low lipophilic antioxidant capacity. In this report, we present a modified 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) method for fruits and compare its performance with the other commonly used antioxidant methods of 2,2'-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP). The antioxidant capacity and reaction kinetics of four phenolic compounds, two antioxidant standards, and five fruits were also investigated. The modified ABTS method prepared at a pH of 4.5 with sodium acetate buffer is highly stable and easily applied to fruit samples as compared to the standard (pH 7.4) version. The measured antioxidant capacity of samples varied with the assay method used, pH, and time of reaction. Traditional antioxidant standards (trolox, ascorbic acid) displayed stable, simple reaction kinetics, which allowed end point analysis with all of assays. Of the phenolic compounds examined, chlorogenic and caffeic acids exhibited the most complex reaction kinetics and reaction rates that precluded end point analysis while gallic acid and quercetin reached stable end points. All fruit extracts exhibited complex and varied kinetics and required long reaction times to approach an end point. Because the antioxidant capacity of fruit extracts is a function of the array of individual antioxidants present, accurate comparisons among fruit samples require that reaction times be standardized and of sufficient length to reach steady state conditions and that more than one assay be used to describe the total antioxidant activity of fruit samples.     

Diversity in Antioxidant Capacity,Phenolic Contents,and Flavonoid Contents of 42 Edible Beans from China

Edible beans are among the most important grain legumes consumed by humans. To provide new information on the antioxidant phenolics of edible beans, the antioxidant capacity, total phenolic content (TPC), and total flavonoid content (TFC) in both soluble and bound fractions of 42 edible beans from China were systematically evaluated, with main phenolic compounds identified and quantified in 10 beans possessing the highest TPC. Edible beans contained a wide range of total antioxidant capacity and TPC generally comparable with common grains, fruits, and vegetables, and their bound fractions had significant antioxidant capacity, TPC, and TFC. Red sword bean was found for the first time to show extremely high total antioxidant capacity (ferrous[II] at 235 ± 13.2 μmol/g and Trolox at 164 ± 10.5 μmol/g) and TPC (1767 ± 58.3 mg of GAE/100 g). Phenolic compounds such as catechin, ferulic acid, gallic acid, p‐coumaric acid, and protocatechuic acid were widely detected in selected beans. A positive correlation was found between antioxidant capacity (ferric‐reducing antioxidant power [FRAP] and Trolox equivalent antioxidant capacity [TEAC] values) and TPC, with correlation coefficient r = 0.974 (FRAP value versus TPC) and r = 0.914 (TEAC value versus TPC). Therefore, beans with high antioxidant capacity and phenolic content can be valuable sources of dietary natural antioxidants for the prevention of oxidative stress‐related chronic diseases.     

Effect of apple cell walls and their extracts on the activity of dietary antioxidants

The effect of dietary fiber in the form of apple cell walls and pectin extracts on natural antioxidants was examined. Cell walls (CW), isolated from apples ( Malus domestica Borkh. cv. "Pacific Rose"), were incubated with ascorbic acid (AA) or quercetin in N-2-hydroxyethylpiperazine- N'-2-ethanesulfonic acid (HEPES) buffer (pH 6.5) at 37 degrees C for 2 h. The resulting supernatants were characterized by a ferric reducing antioxidant power (FRAP) assay and cyclic voltammetry (CV). The experiments were repeated with pectin isolated from the apple cell walls and commercial pectins and showed that polysaccharide preparations stabilized AA effectively but offered little protection against quercetin oxidation. The water-soluble components from cell walls appeared to be responsible for the observed effects of cell-wall polysaccharide preparations on antioxidant activity.     

Influence of processing on quality parameters of strawberries

To determine the effects of different processing steps, such as enzymatic treatment of the mash and pasteurization, on selected quality parameters, strawberries were processed to juices and purees. To identify the processing steps causing the highest losses, samples were taken after each step, and ascorbic acid, total phenols, anthocyanins, and antioxidant capacity were analyzed. To assess the antioxidant capacity, three different methods were applied: the trolox equivalent antioxidant capacity (TEAC), the ferric reducing antioxidant power (FRAP), and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, showing correlation coefficients of 0.889 to 0.948. The antioxidant capacity decreased with processing steps except heat treatment, which partly caused an increase due to the formation of antioxidant active products. The content of ascorbic acid, in comparison to that in the frozen strawberries, decreased significantly during the processing of the fruit to puree by 77%. In the pressed cloudy juices, the loss of ascorbic acid was 37%. The decline of phenolic compounds, measured as total polyphenols and anthocyanins, was smaller (between 30-40%). Pressing and pasteurization were the most critical steps for the decrease of these compounds. The enzymatic treatment of the mash within 90 min supported the release of secondary plant metabolites, while ascorbic acid is reduced up to 20%.     

Polyphenol composition and antioxidant activity in strawberry purees; impact of achene level and storage

In this study the impact of achenes on polyphenolic compounds, ascorbic acids, and antioxidant activities in strawberry purees at production and after storage at 6 and 22 degrees C for 8 and 16 weeks was investigated. Strawberry purees were made from flesh, berry, and achene-enriched homogenate and contained 0, 1.2, and 2.9% achenes, respectively. At production, strawberry purees made from flesh contained more anthocyanins, p-coumaroyl glycosides, and ascorbic acids, whereas increasing achene levels caused increasing levels of ellagic acid derivatives, proanthocyanidins, flavonols, total phenolics (TP), and antioxidant activities. In addition, the anthocyanins, TP, and ferric reducing ability power (FRAP) in purees with more achenes were better retained during storage. Ascorbic acids and anthocyanins declined rapidly during storage, whereas other polyphenols and antioxidant activities were more stable; that is, the contributions from anthocyanins and ascorbic acids to TP and antioxidant activities decreased. The findings that achenes contributed significantly to polyphenol content and stability of strawberry purees may be interesting in a nutritional and, thus, commercial, perspective.     

Use of reference compounds in antioxidant activity assessment

The choice of reference compounds is examined as a "critical control point" of antioxidant activity assessment. Gallic, caffeic, sinapic, uric, and ascorbic acids, isoeugenol, and Trolox were tested using different redox (FRAP, Folin-Ciocalteu) and radical scavenging (DPPH*, ABTS*+, CBA, ORAC) assays. The ability to chelate transition metals was assessed to support some of the findings. Analytes were also tested in liposomes. On the basis of the findings, we do not recommend uric acid (due to solubility constrains) and ascorbic acid (due to fast degradation kinetics) as references. The behavior of the rest of the compounds could not always be attributed to typical structural characteristics. Selection of suitable reference compounds for in vitro antioxidant activity assays is not an easy task to achieve. The choice of reference compounds has to remain at the convenience of the researchers, with regard to the aim of the study.     

Inhibition of low-density lipoprotein oxidation by carnosine histidine

Carnosine is a beta-alanylhistidine dipeptide found in skeletal muscle and nervous tissue that has been reported to possess antioxidant activity. Carnosine is a potential dietary antioxidant because it is absorbed into plasma intact. This research investigated the ability of carnosine to inhibit the oxidation of low-density lipoprotein (LDL) in comparison to its constituent amino acid, histidine. Carnosine (3 microM) inhibited Cu2+-promoted LDL (20 of protein/mL) oxidation at carnosine/copper ratios as low as 1:1, as determined by loss of tryptophan fluorescence and formation of conjugated dienes. Carnosine (6 microM) lost its ability to inhibit conjugated diene formation and tryptophan oxidation after 2 and 4 h of incubation, respectively, of LDL with 3 microM Cu2+. Compared to controls, histidine (3 microM) inhibited tryptophan oxidation and conjugated diene formation 36 and 58%, respectively, compared to 21 and 0% for carnosine (3 microM) after 3 h of oxidation. Histidine was more effective at inhibiting copper-promoted formation of carbonyls on bovine serum albumin than carnosine, but carnosine was more effective at inhibiting copper-induced ascorbic acid oxidation than histidine. Neither carnosine nor histidine was a strong inhibitor of 2,2'-azobis(2-amidinopropane) dihydrochloride-promoted oxidation of LDL, indicating that their main antioxidant mechanism is through copper chelation.     

The role of oxylipins and antioxidants on off-flavor precursor formation during potato flake processing

The impact of processing on nonenzymatic antioxidant degradation and lipid oxidation leading to off-flavor development in potato flakes during storage was investigated. Lipoxygenase activity measurements in parallel with the analysis of lipid oxidation products (oxylipins) profiles using HPLC showed that the processing conditions used inhibited efficiently enzymatic lipid oxidation. However, nonenzymatic lipid oxidation products were found throughout processing and in fresh potato flakes. Furthermore, these autoxidative processes cannot be inactivated by the main endogenous nonenzymatic antioxidants in potato tubers (ascorbic acid, phenolic compounds and carotenoids), as these antioxidants are degraded during processing. Indeed, leaching and thermal treatments taking place during processing lead to a decrease of about 95%, 82% and 27% in the content of ascorbic acid, phenolic compounds and carotenoids, respectively. Therefore, storage is a critical step to prevent off-flavor development in potato flakes. Specific attention has thus to be paid on the use of efficient exogenous antioxidants as well as on storage conditions.     

Repeated red wine consumption and changes on plasma antioxidant capacity and endogenous antioxidants (uric acid and protein thiol groups)

This study aims to ascertain the in vivo antioxidant properties of red wine by determining how it affects antioxidant biomarkers (plasma antioxidant capacity (PAC) and endogenous antioxidants such as uric acid or protein thiol groups). Antioxidant biomarkers have been assessed in eight healthy human volunteers after repeated intakes of 300 mL of red wine every day for a week. During this intervention period, volunteers followed a low phenolic diet designed to prevent the phenolic compounds in wine from interfering with the phenolics from other foods or beverages. This diet was followed throughout the week that the volunteers drank wine and for another control week when they drank water. Biomarkers were determined before the subjects taking part in the study started the intervention period with red wine (Monastrell variety) and 1, 3, and 7 days after. PAC was evaluated by the Ferric Reducing Ability of Plasma assay (FRAP), and the Oxygen Radical Absorbance Capacity assay using fluorescein (ORAC-FL). In addition, the concentrations of endogenous antioxidants such as uric acid, albumin, bilirubin, and protein thiol groups were analyzed. The FRAP method shows that PAC increased after the week of wine consumption but decreased after the week without wine consumption. The uric acid concentration did not show any changes that were significantly different from our results in acute wine intake studies. Protein thiol groups decreased significantly (p < 0.05) with the low phenolic diet, but this decrease was not statistically significant if the diet was taken with red wine (p < 0.05).     

Assessing the antioxidant activity of melanoidins from coffee brews by different antioxidant methods

Antioxidant activity of instant coffees produced from the same green coffee beans roasted at three different degrees was analyzed. Coffee melanoidins were obtained by ultrafiltration (10 kDa cutoff) and subsequent diafiltration. Pure melanoidins were isolated from melanoidins after overnight incubation in 2 M NaCl and then ultrafiltered. Filtrates, corresponding to the low molecular weight (LMW) fraction noncovalently linked to the melanoidin skeleton, were also preserved. Antioxidant activity of coffee brews (CB), melanoidins (M), pure melanoidins (PM), and bounded melanoidin compounds (BMC) were tested using the 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and ferric reducing power (FRAP) methods. The correlation between the different methods was studied. The higher contribution of melanoidins to the total antioxidant activity of coffees was shown to be caused by the LMW compounds linked noncovalently to the melanoidin skeleton, as data from BMC confirmed. CB, M, and BMC fractions exert the highest antioxidant activity in aqueous media, whereas PM was not dependent on the reaction media. The highest correlation was found between DPPH and FRAP methods.     

Evolution of antioxidant capacity during storage of selected fruits and vegetables

Interest in the consumption of fresh fruits and vegetables is, to a large extent, due to its content of bioactive nutrients and their importance as dietary antioxidants. Among all of the selected fruits and vegetables, strawberries and black grapes have relatively high antioxidant capacities associated with high contents of total phenolic compounds, ascorbic acid, and flavonols. More interesting, the results of this study indicated that in most fruits and vegetables storage did not affect negatively the antioxidant capacity. Better, in some cases, an increase of the antioxidant capacity was observed in the days following their purchase, accompanied by an increase in phenolic compounds. In general, fruits and vegetables visually spoil before any significant antioxidant capacity loss occurs except in banana and broccoli. When ascorbic acid or flavonoids (aglycons of flavonols and anthocyanins) were concerned, the conclusions were similar. Their content was generally stable during storage.     

Antioxidant capacity of plasma after red wine intake in human volunteers

Plasma antioxidant capacity (AC) has been assessed in eight healthy human volunteers after wine intake. Analytical methods used for evaluating AC included the ferric reducing ability of plasma (FRAP) and oxygen radical absorbance capacity using two different fluorescent probes, beta-phycoerythrin (ORAC-PE) and fluorescein (ORAC-FL). In addition, the concentrations of endogenous antioxidants such as uric acid, albumin, and bilirubin were determined. The suitability of analytical methods was evaluated with two different biological matrixes: plasma and serum. Plasma AC was determined before ingestion of 300 mL of red wine (baseline value) and 30, 55, and 120 min afterward. Maximum average increase in AC values was reached at 55 min. Plasma AC determined by ORAC-PE at time zero was significantly correlated with albumin concentration. Plasma AC determined with FRAP at time zero is well correlated with uric acid. Moreover, a good linear correlation was found between uric acid concentration and AC determined by FRAP in each volunteer. The maximum concentration of uric acid occurred after 55 min. Uric acid increase accounts for a nonnegligible part in FRAP values and must be evaluated when using this method for assessing AC in plasma.     

Analysis of antioxidant activities of common vegetables employing oxygen radical absorbance capacity (ORAC) and ferric reducing antioxidant power (FRAP) assays: a comparative study

A total of 927 freeze-dried vegetable samples, including 111 white cabbages, 59 carrots, 51 snap beans, 57 cauliflower, 33 white onions, 48 purple onions, 130 broccoli, 169 tomatoes, 25 beets, 88 peas, 88 spinach, 18 red peppers, and 50 green peppers, were analyzed using the oxygen radical absorption capacity (ORAC) and ferric reducing antioxidant capacity (FRAP) methods. The data show that the ORAC and FRAP values of vegetable are not only dependent on species, but also highly dependent on geographical origin and harvest time. The two antioxidant assay methods, ORAC and FRAP, also give different antioxidant activity trends. The discrepancy is extensively discussed based on the chemistry principles upon which these methods are built, and it is concluded that the ORAC method is chemically more relevant to chain-breaking antioxidants activity, while the FRAP has some drawbacks such as interference, reaction kinetics, and quantitation methods. On the basis of the ORAC results, green pepper, spinach, purple onion, broccoli, beet, and cauliflower are the leading sources of antioxidant activities against the peroxyl radicals.     

Phenol antioxidant quantity and quality in foods: fruits.

The free and bound phenols have been measured in 20 fruits commonly consumed in the American diet. Phenols were measured colorimetrically using the Folin-Ciocalteu reagent with catechin as the standard after correction for ascorbic acid contribution. On a fresh weight basis, cranberry had the highest total phenols, and was distantly followed by red grape. Free and total phenol quality in the fruits was analyzed by using the inhibition of lower density lipoprotein oxidation promoted by cupric ion. Ascorbate had only a minor contribution to the antioxidants in fruits with the exception of melon, nectarine, orange, white grape, and strawberry. The fruit extracts' antioxidant quality was better than the vitamin antioxidants and most pure phenols, suggesting synergism among the antioxidants in the mixture. Using our assay, fruits had significantly better quantity and quality of phenol antioxidants than vegetables. Fruits, specifically apples and cranberries, have phenol antioxidants that can enrich lower density lipoproteins and protect them from oxidation. The average per capita consumption of fruit phenols in the U.S. is estimated to be 255 mg/day of catechin equivalents.