Evaluation of spectrophotometric methods for antioxidant compound measurement in relation to total antioxidant capacity in beverages

The validity of different colorimetric methods used to quantify various families of antioxidant compounds was evaluated with standard compounds. The colorimetric tests for global evaluation of flavonoids, anthocyans, and flavanols were found generally unreliable, as reactions could be different for individual compounds within a family (anthocyanins or flavonols or flavan-3-ols) and not specific to one family. In the flavonoid test, for example, flavonols reacted very well, anthocyanins did not react, and flavanons reacted only slightly. The same methods were applied also to beverages known for their antioxidant content (apple, orange, grape, and vegetable juices, ice tea, and red wine) and the data were compared with the results of HPLC analysis of specific compounds. The values obtained in a colorimetric test were generally higher than the sum of the values obtained for the corresponding individual compounds by HPLC analysis, mainly because other compounds can interfere with the colorimetric tests. For example, in wine, anthocyanin concentrations obtained by colorimetric test were 9068 ± 1407 μmol/100 ml (mean ± SEM), higher than the sum of the six main anthocyanidins detected by HPLC, only 41 μmol/100 ml. The relative antioxidant capacity values determined for beverages on the basis of colorimetric tests could exceed by far the values previously measured in radical-scavenging tests (for instance, the antioxidant capacity attributable to anthocyans in wine on the basis of the colorimetric test was 50 times higher than the total antioxidant capacity measured by the ORAC assay). In conclusion, colorimetric tests for flavonoids, anthocyans, and flavanols appeared generally unreliable for estimating their content and thus the antioxidant capacity reliable to these compounds.     

Antioxidant compounds and antioxidant capacity of Peruvian camu camu (Myrciaria dubia (H.B.K.) McVaugh) fruit at different maturity stages

The antioxidant capacities of ascorbic acid and phenolic compounds present in camu camu fruit were screened during ripening. Ascorbic acid decreased, and anthocyanin, flavonol and flavanol contents, and DPPH antioxidant capacity increased during ripening. Antioxidant compounds from camu camu were fractionated in two fractions: an ascorbic acid-rich fraction (F-I) and a phenolics-rich fraction (F-II). F-I was the major contributor to the DPPH antioxidant capacity (67.5–79.3%) and F-II played a minor role (20.7–32.5%). A total of 30 different phenolic compounds were detected by HPLC-PAD. The presence of catechin, delphinidin 3-glucoside, cyanidin 3-glucoside, ellagic acid and rutin was elucidated. Other phenolic compounds, such as flavan-3-ol, flavonol, flavanone and ellagic acid derivatives, were also present. For the three ripening stages the flavan-3-ols and ellagic acid group were the most representative phenolic compounds in this fruit. Acid hydrolysis of F-II revealed the presence mainly of gallic and ellagic acids, suggesting that camu camu fruit possesses important quantities of hydrolysed tannins (gallo- and/or ellagitannins). These results confirm that camu camu fruit is a promising source of antioxidant phenolics.     

Determination of carotenoids in Dunaliella salina cultivated in Taiwan and antioxidant capacity of the algal carotenoid extract

A simple HPLC method with good separation efficiency was developed to determine all-trans and cis forms of carotenoids in Dunaliella salina cultivated in Taiwan. The analysis used a C30 column (250 × 4.6 mm, 5 μm) and an isocratic solvent system (flow rate = 1 mL/min) mixing methanol–acetonitrile–water (84/14/2, v/v/v) and methylene chloride, (75/25, v/v). Carotenoids were detected at 450 nm. Moreover, the antioxidant capacities of the algal carotenoid extract were also evaluated with Trolox equivalent antioxidant capacity (TEAC) assay, reducing power and 2,2-diphenyl-2-picrylhydrazyl hydrate (DPPH) radical scavenging assay. Results showed that 7 carotenoids in the algal extract could be separated simultaneously within 30 min and the total amount of them was 290.77 mg/g algae. The contents of all-trans-β-carotene and 9- or 9′-cis-β-carotene, the major carotenoids in the algae, were 138.25 and 124.65 mg/g algae, respectively. The contents of all-trans-lutein, all-trans-zeaxanthin, 13- or 13′-cis-β-carotene, all-trans-α-carotene and 9- or 9′-cis-α-carotene were 6.55, 11.27, 4.95, 2.69, and 2.41 mg/g algae, respectively. The algal carotenoid extract had significantly higher antioxidant activity than all-trans forms of α-carotene, β-carotene, lutein and zeaxanthin in all antioxidant assays. The cis forms of carotenoids, especially 9- or 9′-cis-β-carotene, might play crucial roles for the antioxidant capacities of the algal extract.     

Polyphenols and antioxidant capacity of Phaseolus vulgaris stored under extreme conditions and processed

Phaseolus vulgaris is a foodstuff that not only supplies nutrients, but also bioactive compounds with antioxidant capacity; however, bad post-harvest storage or processing can diminish the antioxidant capacity. In this work, the effect of storage under extreme conditions, of thermal processing and of fermentation on the content of phenolic compounds and their antioxidant capacity (AA) in beans of just harvested P. vulgaris were evaluated. Combination of temperatures of 30 and 50 °C and relative humidity of 11% and 80% for 150 days were evaluated. Extreme storage conditions drastically decreased the AA, demonstrating that post-harvest storage at low temperature and humidity preserves the antioxidant capacity of the beans. Regarding processing, cooking decreased the scavenging effect and the AA by 63.5% and 56.8%, while fermentation decreased them by only 14% and 22%, respectively. It was concluded that the loss of antioxidant capacity due to heating is less in fermented beans.     

Investigation of the effects of different organosolv pulping methods on antioxidant capacity and extraction efficiency of lignin

Different organosolv pulping methods delignified raw materials (Acanthopanax senticosus remainder) were investigated, including acetone-acetic acid-water method (8/1/1, v/v/v), 1,4-butanediol-acetic acid-water method (8/1/1, v/v/v), ethanol-acetic acid-water method (8/1/1, v/v/v) and acetic acid-water method (4/1, v/v). Lignin extraction efficiency and antioxidant capacity from different organosolv methods were compared.The results showed that the lignin extraction efficiency of the 1,4-butanediol-acetic acid-water method was the highest (39%) among methods of this paper. The lignin from acetic acid-water method was the most effective free radical scavengers, whose 1,1-di-phenyl-2-picrylhydrazyl (DPPH) radical scavenging capacity (IC₅₀) was about 0.6587 mg/ml.Acanthopanax senticosus remainder (raw materials) and pulps were characterised by scanning electron microscopy (SEM) and pulps strength properties. All lignin samples were analysed by Fourier-transform Infrared Spectroscopy (FTIR). The results showed that organosolv pulping process had large influence on pulps morphology, pulps strength properties and lignin structure.     

Chemical profile and antioxidant capacities of tart cherry products

The levels of anthocyanins and other flavonoids, as well as melatonin, in various tart cherry products (frozen and dried cherries, powders from individually quick frozen (IQF) cherry and juice concentrate) from two tart cherry cultivars, ‘Montmorency’ and ‘Balaton’ were analysed comparatively by HPLC and electrospray mass spectrometry (EMS). Our results show that the major anthocyanin compound in these two tart cherry cultivars is cyanidin 3-glucosylrutinoside, followed by cyanidin 3-rutinoside, cyanydin sophoroside, and peonidin 3-glucoside. Studies on antioxidant activities (total antioxidant status assay) of crude extracts of ten tart cherry products show that these products preserve their antioxidant capacities after processing and storage. We have also compared the antioxidant activities of several single constituents that are present in tart cherry. When TEAC (trolox equivalent antioxidant capacity) values were evaluated conceptually against the cherry phytochemical profile, cyanidin and its derivatives were found to be the significant contributors to the antioxidant systems of tart cherries. It was shown that standard compounds with common aglycon moieties show similar antioxidant activities.     

Enhanced antioxidant activity of Monascuspilosus fermented products by addition of ginger to the medium

The fermented products of Monascus sp. are known for their antihypercholesterolaemic effects, however, their antioxidant activities are different from those of many plant-derived foods. To evaluate the effect of ginger addition into the medium on the antioxidant activity of Monascuspilosus fermented products, we cultured uninoculated PDB medium (PDB), inoculated PDB medium (MP), uninoculated ginger-containing medium (PDBG), and inoculated ginger-containing medium (MPG). The broth and mycelia were collected, freeze-dried, and extracted to evaluate their free radical scavenging activities, inhibition of peroxidation, phenolic content, inhibition of DNA damage, cellular antioxidant activity, and expression of the antioxidant enzymes. The results showed that MPG had significantly higher antioxidant activity than PDB, MP, and PDBG at all fermentation time points. Moreover, the fermentation process significantly increased the antioxidant activities of MPG. After the inherent level of antioxidant capacity was increased, the modified M. pilosus fermented product demonstrated a higher anti-atherosclerotic value than the unmodified product.     

Changes in bioactive compounds and antioxidant capacity of fresh-cut cashew apple

In this study, ascorbic acid, total polyphenols and proanthocyanidins of fresh-cut cashew apple were quantified. Antioxidant capacity was determined in whole juice and in polyphenols extracts by three methods: 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP) and ??-carotene bleaching. Effect of cutting and storage for 24 h at 2 °C, 27 °C and 40 °C on these compounds were also evaluated. Cashew apple presented 163 mg of ascorbic acid per 100 g of fresh weight (FW). Soluble and hydrolysable polyphenols contents were 12.79 mg GAE/100 g FW and 18.53 mg GAE/100 g FW and proanthocyanidins were 9.27 mg/100 g FW. Antioxidant capacity of juice and polyphenols extract was high for DPPH method. Storage temperatures affected bioactive compounds on cut cashew apple. The content of ascorbic acid decreased in all temperatures. Proanthocyanidins were more sensitive to 40 °C than to other temperatures. The content of polyphenols and antioxidant capacity of juice by DPPH assay did not change. However, the reducing power was lower in samples kept at high temperatures. A strong positive correlation between ascorbic acid and FRAP (r = 0.99) and a negative correlation between DPPH and FRAP (r = − 0.79) were observed. No correlations were found between polyphenols and antioxidant capacity indicating the importance of phenolic composition in the extracts. The results confirm the importance of temperature and injury on the quality of cashew apple.     

Evaluation of the antioxidant activity of three microalgal species for use as dietary supplements and in the preservation of foods

The antioxidant activity of the microalgal ethanolic extracts of Porphyridium cruentum, Phaeodactylum tricornutum and Chlorella vulgaris was determined by means of the β-carotene–linoleate model system. The results show that the activity of C. vulgaris extract was higher than those obtained for the other microalgal extracts tested and for the synthetics BHA (butylated hydroxyanisole), and BHT (butylated hydroxytoluene). In addition, the major constituents present in the ethanolic extracts of the three microalgae species were analyzed by means of GC and GC–mass spectrometry. The results showed that the tested microalgae may be an important source of natural antioxidants, as an alternative to higher plants or the production by chemical synthesis.     

Chemical characterization and antioxidant capacity of berries from Clidemia rubra (Aubl.) Mart. (Melastomataceae)

The flora of Latin America attracts gaining interest as it provides a plethora of still unexplored or under-utilized fruits that can contribute to human well-being due to their nutritional value and their content of bioactive compounds. Clidemia rubra (Aubl.) Mart. is a shrub belonging to the family of the Melastomataceae that grows preferably in a tropical climate. This paper comprises a nutritional characterization of the berries from Clidemia rubra and provides data on the phenolic compounds as well as the antioxidant capacity of the fruit. Findings in macronutrients like protein, carbohydrates, and fat were comparable to that of common berry fruits. Clidemia rubra berries seemed to be a good source for dietary fibers and some minerals (Ca, Mn, and Zn). In contrast, contents of titratable acids and ascorbic acid were low. The polyphenolic profile was determined by using HPLC-MS/MS in comparison to standard compounds. Noteworthy amounts of cyanidin 3-O-rutinoside (39.43 ± 1.66 mg/100 g fresh weight (FW)), delphinidin 3-O-rutinoside (23.74 ± 1.18 mg/100 g FW), cyanidin 3-O-glucoside (11.68 ± 0.56 mg/100 g FW), and delphinidin 3-O-glucoside (6.08 ± 0.35 mg/100 g FW) were found. Non-anthocyanin phenolic constituents were phenolic acids (gallic, protocatechuic, p-hydroxy-benzoic, vanillic, and caffeic acid), flavan-3-ols (epigallocatechin, epigallocatechin gallate, and epicatechin gallate), and 11 different myricetin and quercetin derivatives of which quercetin 3-O-arabinoside (5.26 ± 0.16 mg/100 g FW) and quercetin 3-O-rhamnoside (5.06 ± 0.08 mg/100 g FW) were dominating. Anthocyanins and ascorbic acid were mainly responsible for the antioxidant capacity of Clidemia rubra berries assessed with the total oxidant scavenging capacity (TOSC) assay.     

Polysaccharides from Turbinaria conoides: Structural features and antioxidant capacity

Since reactive oxygen species (ROS) are involved in initiating and promoting several diseases such as cancer and cardiovascular events, this study was designed to evaluate the antioxidant activity of sulphated polysaccharides purified from Turbinaria conoides. A fucoidan (AF3), one alginic acid (B) and a neutral glucan (C1F2) were isolated. The glucan is made up of β-(1 → 3)-linked glucopyranosyl residues and has a molecular mass of 5 kDa. The alginic acid contains equimolar proportions of guluronic and mannuronic acid residues. The fucoidan is highly branched and has an average molecular mass of 50 kDa. The in vitro antioxidant capacity of the fractions was studied by ferric reducing antioxidant power (FRAP) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) assays. The fucoidan (AF3) showed the highest potential followed by the alginate (B) and laminaran (C1F2). Taken together, these findings demonstrate that these polysaccharides could be used as natural antioxidants by the food industry.     

The increase in antioxidant capacity after wounding depends on the type of fruit or vegetable tissue

Wounding of fresh produce may elicit an increase in antioxidant capacity associated with wound-induced phenolic compounds. However, there have been no reports on the wounding response of different types of fresh produce. Changes in antioxidant capacity, total soluble phenolics, ascorbic acid, total carotenoids and total anthocyanins were evaluated after wounding in zucchini, white and red cabbage, iceberg lettuce, celery, carrot, parsnips, red radish, sweetpotato and potatoes. Phenolic changes ranged from a 26% decrease to an increase up to 191%, while antioxidant capacity changes ranged from a 51% decrease to an increase up to 442%. Reduced ascorbic acid decreased up to 82%, whereas the changes in anthocyanins and carotenoids were less evident. In general, the wound response was dependent on the type of tissue and influenced by the initial levels of reduced ascorbic acid and phenolic compounds. Wounding may increase the antioxidant content towards the development of selected healthier fresh-cut produce.     

Organic acids,antioxidant capacity,phenolic content and lipid characterisation of Georgia-grown underutilized fruit crops

Four underutilized Georgia-grown fruit crops, namely loquat (Eriobotrya japonica), mayhaw (Crataegus sp.), fig (Ficus carica), and pawpaw (Asimina triloba), and their leaves were analysed for total polyphenols by Folin–Ciocalteau method, and antioxidant capacity by ferric-reducing antioxidant power (FRAP) and Trolox-equivalent antioxidant capacity (TEAC) assays. Organic acids and phenolic compounds were identified by RP-HPLC. For lipid profile, fruits were separated into two fractions – seed and fruit (i.e., without seed); lipid was extracted using the Folch method and analysed for fatty acids, phytosterols, tocopherols, and phospholipids. The major organic acid identified in all samples was malic acid (177–1918 mg/100 g FW). The predominant phenolic acids in all the fruits were gallic (1.5–6.4 mg/100 g FW) and ellagic (0.2–33.8 mg/100 g FW), and the most abundant flavonoid was catechin (12.2–37.8 mg/100 g FW). Total lipid content varied from 0.1% in mayhaw fruit to 21.5% in pawpaw seed. Linoleic acid was the predominant fatty acid in all of the samples (28.2–55.7%).     

Flow-sonochemiluminescence method for antioxidant capacity analysis

A flow-sonochemiluminescence (SCL) method that offers a fast and simple alternative to the antioxidant capacity analysis is described. Reactive oxygen species (ROS) is generated reagentlessly and in situ in pH 7.4 phosphate buffer saline by ultrasound irradiating. Luminol reacts with the ROS to give SCL. The presence of antioxidant quenches the ROS, which in turn quenches the SCL. By quantifying SCL inhibition, the antioxidant capacity measurement can be achieved. Antioxidants including uric acid, l-ascorbic acid, tannic acid, l-cysteine, citric acid, salicylic, dl-sorbol and glutathione, are selected to test the presented method. Linear relationships between the quenched SCL intensity and antioxidant concentration, IC₅₀ (half-inhibition concentration) of each antioxidant are obtained. Antioxidant capacities of six vegetables extracts (onion, potato, cabbage, carrot, cucumber and spinach) are evaluated. Under the optimized experiment condition, a high detection frequency of 590 h⁻¹ is achieved. Intra- and inter-assay variability investigation shows relatively good precision of this flow-SCL method.     

Varieties of antioxidant and antibacterial properties of Ecklonia stolonifera and Ecklonia kurome products harvested and processed in the Noto peninsula,Japan

Ecklonia stolonifera and Ecklonia kurome are traditional edible brown algae in the fisheries towns in Far East Asia. In the Noto Peninsula area, Ishikawa, Japan, both the Ecklonia are called ‘kajime’ and people believe that the algae improve the property of blood. To determine the varieties of antioxidant and antibacterial properties E. stolonifera and E. kurome products, assays for total phenolic content and antioxidant activities, including DPPH radical-scavenging activity, superoxide anion radical-generated by non-enzymatic system, ferrous-reducing power and WST-8, a tetrazolium salt, redox activity of four dried and two boiled E. stolonifera, and four dried and two raw E. kurome preparations were tested in this study. Furthermore, antibacterial activity of the products was tested. Though the total phenolic content, the antioxidant activity and the antibacterial activities of E. stolorifera and E. kurome products were high, these properties were varied by manufacturers or each product. Especially, two dried and two boiled products of E. stolonifera showed low activities. The results of this study indicate that the contents of functional compounds and their activities were affected or decreased greatly by the processing method.     

Total phenolics,ascorbic acid,and antioxidant capacity of noni (Morinda citrifolia L.) juice and powder as affected by illumination during storage

Total phenolics, ascorbic acid, and antioxidant capacity of noni (Morinda citrifolia L.) juice and powder were determined during storage at 24 °C. After 2 weeks of storage, illuminated noni juice lost 32% of total phenolics, 89% of ascorbic acid, and 46–65% of antioxidant capacity—about 8%, 22%, and 9–15% more than unilluminated juice. Both illuminated and unilluminated juice lost 97% of ascorbic acid by 4 weeks. The difference in antioxidant characteristics between illuminated and unilluminated juice became insignificant at 12 weeks. After 12 weeks, illuminated noni powder lost 21% of total phenolics, 17% of ascorbic acid, and 23–36% of antioxidant capacity—about 13%, 4%, and 7–19% more than the unilluminated powder. Noni powder in brown bottles retained antioxidant characteristics significantly greater than that in clear bottles. Protection from light effectively reduced degradation of antioxidant characteristics of noni juice for only 2 weeks but those of noni powder for at least 3 months.     

Peanut antioxidants: Part 1. Genotypic variation and genotype-by-environment interaction in antioxidant capacity of raw kernels

The potential to select for antioxidant traits in peanut (Arachis hypogaea L.) kernels by conventional plant breeding was investigated by oxygen radical absorbance capacity (ORAC) assay of (a) 32 full-season maturity genotypes grown at Kairi Research Station in 2008/09; and (b) a subset of ten genotypes with diverse antioxidant capacities grown in five different production regions in Queensland in 2009/10 and 2010/11. There were significant levels of variation of more than 25% relative standard deviation (RSD) in antioxidant capacity among the 32 tested genotypes; and that genotype, environment, and genotype-by-environment (G × E) interaction all significantly (P < 0.05) affected trait expression. Analysis of the G × E data confirmed the heritability of antioxidant capacity, which has not been previously quantified, whereby genotype explained 44% of the phenotypic variation on a plot basis and 82% on an entry mean basis. This indicated that there was substantial genetic control of antioxidant capacity in peanut kernels, but also that it will be important to characterize environmental interaction to enable plant/seed selection in the Breeding Program. The research also highlighted the utility of the ORAC assay for screening germplasm and provided practical advice on design of varietal assessments using antioxidant screenings.     

Influence of apple purée preparation and storage on polyphenol contents and antioxidant activity

The polyphenolic components of Idared and Shampion apple purées were determined by HPLC; chlorogenic acid was the most abundant acid (20.0 mg/100 g in microwaved Idared); other polyphenols identified in high concentrations included (−)-epicatechin, procyanidin B1 and B2; quercetin and cyanidin glycosides were found in minor concentrations. The Shampion purées had higher total phenolics (142 mg/100 g) and procyanidin B2 concentrations (17.3 mg/100 g) than the Idared ones, and polymeric procyanidins represented 41% of all polyphenols. Antioxidant capacities of the samples, determined by spectrophotometric methods and EPR spectroscopy, correlated well with the concentration of polyphenols. The antioxidant properties could be better represented by EPR than by UV–vis measurements. The latter require transparent (clear) samples whereas EPR can be a method of choice in the assessment of radical-scavenging activity of dense and cloudy apple purées. Our results support the putative high antioxidant value of apple purées and define their capacity in terms of the major constituents. Apple purées are a rich source of natural antioxidants, especially of chlorogenic acid and procyanidins.