Antioxidant activity in fruits and leaves of blackberry, raspberry, and strawberry varies with cultivar and developmental stage

Fruits and leaves from different cultivars of thornless blackberry (Rubus sp.), red raspberry (Rubus idaeus L.), black raspberry (Rubus occidentalis L.), and strawberry (Fragaria x ananassa D.) plants were analyzed for total antioxidant capacity (oxygen radical absorbance capacity, ORAC) and total phenolic content. In addition, fruits were analyzed for total anthocyanin content. Blackberries and strawberries had the highest ORAC values during the green stages, whereas red raspberries had the highest ORAC activity at the ripe stage. Total anthocyanin content increased with maturity for all three species of fruits. Compared with fruits, leaves were found to have higher ORAC values. In fruits, ORAC values ranged from 7.8 to 33.7 micromol of Trolox equivalents (TE)/g of fresh berries (35. 0-162.1 micromol of TE/g of dry matter), whereas in leaves, ORAC values ranged from 69.7 to 182.2 micromol of TE/g of fresh leaves (205.0-728.8 micromol of TE/g of dry matter). As the leaves become older, the ORAC values and total phenolic contents decreased. The results showed a linear correlation between total phenolic content and ORAC activity for fruits and leaves. For ripe berries, a linear relationship existed between ORAC values and anthocyanin content. Of the ripe fruits tested, on the basis of wet weight of fruit, cv. Jewel black raspberry and blackberries may be the richest source for antioxidants. On the basis of the dry weight of fruit, strawberries had the highest ORAC activity followed by black raspberries (cv. Jewel), blackberries, and red raspberries.     

Antioxidant capacity in cranberry is influenced by cultivar and storage temperature

Ten cranberry (Vaccinium macrocarpon Aiton) cultivars were evaluated for oxygen radical absorbance capacity (ORAC), anthocyanins, and total phenolics contents after three months of storage at 0, 5, 10, 15, and 20 degrees C. The antioxidant capacity of cranberry was affected by cultivars and storage temperatures. Among the 10 cranberry cultivars used in this study, Early Black, Crowley, and Franklin had higher antioxidant capacities than the other cultivars. ORAC values, anthocyanins, and total phenolics contents increased during storage. The highest increases in antioxidant activity, anthocyanin, and phenolics contents occurred at 15 degrees C storage. Fruit stored at 20 degrees C had lower ORAC values than those stored at 15 degrees C. A positive relationship existed between ORAC values and anthocyanin or phenolic content in all 10 cranberry cultivars at different storage temperatures.     

Antioxidant capacity and phenolic content of spinach as affected by genetics and growing season

Total phenolic content and antioxidant capacity of 11 commercial cultivars and 15 advanced breeding lines of spinach were determined over two growing seasons known to vary in biotic and abiotic stresses. Flavonoid composition and content of fall-grown commercial cultivars and advanced breeding lines were also determined. Over-winter spinach, which was planted in late fall and harvested in the spring, had much higher levels of total phenolics and antioxidant capacity than spinach planted in early fall and harvested in late fall, indicating that growing conditions, as well as biotic and abiotic stresses, influenced phenolic metabolism. Genotype also appeared to play an important role in affecting phenolic metabolism and antioxidant capacity in spinach. Advanced breeding lines of spinach, which show increased disease resistance, had higher levels of total phenolics, individual and total flavonoids, and antioxidant capacity than commercial cultivars. Our results indicate that plant breeders can select for increased phenolic content to increase antioxidant capacity in spinach cultivars, or the crops can be grown in different seasons or under certain stress conditions to elevate levels of antioxidants.     

Biochemical profiling and chemometric analysis of seventeen UK-grown black currant cultivars

Black currant fruits are recognized as being an important dietary source of health-related compounds, such as anthocyanins and ascorbic acid. In the present study, the biochemical composition (viz., nonstructural carbohydrates, individual anthocyanins, total anthocyanins, total phenolics, and organic acids, including ascorbic acid) from 17 UK-grown black currant cultivars was analyzed. Berry composition was significantly affected by genotype. Nonstructural carbohydrates ranged from 85.09 to 179.92 mg g(-1) on a fresh weight (FW) basis, while concentration for organic acids ranged from 36.56 to 73.35 mg g(-1) FW. Relative concentrations of cyanidin 3-glucoside, cyanidin 3-rutinoside, delphinidin 3-glucoside and delphinidin 3-rutinoside were 3.1-7.9%, 35.4-47.0%, 7.6-12.5% and 36.9-50.9%, respectively. Differences in the biochemical profile among cultivars were emphasized by principal component analysis (PCA) and hierarchical cluster analysis (HCA). PCA was able to discriminate between cultivars, especially on the basis of health-related compounds. Initial exploration revealed that individual anthocyanins, total phenolics, and ascorbic acid could be used to characterize and classify different cultivars. HCA showed that the biochemical composition of the different cultivars was related to parentage information.     

Antioxidant activity of black currant anthocyanin aglycons and their glycosides measured by chemiluminescence in a neutral pH region and in human plasma

The antioxidant activity of nine anthocyanin glycosides was measured in a neutral pH region using a chemiluminescence (CL) emission system in the presence of an H(2)O(2)-acetaldehyde system, and the intensities were found to be affected by three factors, pH value and both moieties of the aglycon and C-3 sugar. With an increase in pH from 4.0 to 9.0, the CL intensities increased from pH 5.0, reached their maxima at pH 6.0-7.0, and decreased at pH 9.0. Comparison of the intensities among the 3-glucosides with five different aglycons and the 3-glycosides with three different sugar moieties at C-3 showed that their strongest intensities were given by the delphinidin aglycon and 3-rutinosyl moiety, respectively. Monitoring of the CL intensity of human blood plasma for 8 h after oral administration of black currant anthocyanins (BCA) showed a rapid increase until 2 h, and a significant difference (P < 0.05) was recognized at 1-8 h.     

Characterization and fate of black currant and bilberry flavonols in enzyme-aided processing

The fate of black currant ( Ribes nigrum L.) and bilberry ( Vaccinium myrtillus L.) flavonols in enzyme-aided processing was studied. The flavonols were quantified and characterized by high-performance liquid chromatography equipped with a diode array detector and an electrospray ionization mass spectrometer. A tentative identification for 14 black currant and 19 bilberry flavonols is presented representing 11 previously unpublished conjugates. For the first time in any berry, the presence of laricitrin conjugates is reported. The enzyme-aided processing affected the flavonol extractability, elevating the yield in juices and decreasing that in press residues. Importantly, no significant loss of the berry flavonols was observed during the experiments, although some hydrolysis of flavonol conjugates was recorded. To maximize the effect on flavonol extractability, higher enzyme dosages were needed for black currants than for bilberries. The data show that the flavonol extractability and hydrolysis are dependent on the texture of raw material, the glycosylation pattern of the conjugates, and the activity profile of the enzyme preparation.     

Effect of processing of black currant press-residue on polyphenol composition and cell proliferation

The press-residue of black currants provides a good source of phenolic antioxidants. The purpose of this study was to optimize the extraction of phenolic compounds from the press-residue by use of extraction conditions compatible with food use. The effects of temperature, extraction duration, and use of ultrasound-assisted extraction on the juice yield, total phenolics (TP), and anthocyanin content of aqueous extracts were studied. Within the variables and response factors tested, the optimal conditions were a 15 min extraction at 90 °C. No significant effect from ultrasound-assisted extraction was found. The composition of anthocyanins and polyphenols was highly dependent on the extraction temperature. The percentage contribution of delphinidin- and cyanidin-3-rutinoside to TP had a negative linear correlation with temperature, while delphinidin- and cyanidin-3-glucoside had a positive linear correlation with temperature, with a maximum amount obtained at 80 °C and 55 °C, respectively. Furthermore, extracts obtained at higher temperatures showed a stronger inhibition of proliferation of Caco-2, HT-29, and HCT 116 cells than extracts obtained at lower temperatures. This may be due to the decomposition of complex polyphenols at higher temperatures, making them more accessible to the cells.     

Absorption and excretion of black currant anthocyanins in humans and watanabe heritable hyperlipidemic rabbits

Anthocyanins are thought to protect against cardiovascular diseases. Watanabe heritable hyperlipidemic (WHHL) rabbits are hypercholesterolemic and used as a model of the development of atherosclerosis. To compare the uptake and excretion of anthocyanins in humans and WHHL rabbits, single-dose black currant anthocyanin studies were performed. Procedures for workup and analyses of urine and plasma samples containing anthocyanins were developed with high recoveries (99 and 81%, respectively) and low limits of quantification (> or =6.6 and > or =1.1 nM, respectively). The excretion and absorption of anthocyanins from black currant juice were found to be within the same order of magnitude in the two species regarding urinary excretion within the first 4 h (rabbits, 0.035%; humans, 0.072%) and t(max) (rabbits, approximately 30 min; humans, approximately 45 min). A food matrix effect was detected in rabbits, resulting in the absorption of a higher proportion of the anthocyanins from black currant juice than from an aqueous citric acid matrix. In humans the absorption and urinary excretion of anthocyanins from black currant juice were found to be proportional with dose and not influenced by the ingestion of a rice cake. In both species a larger proportion of the anthocyanin rutinosides than of the glucosides was absorbed, whereas the structure of the aglycon had no influence on the absorption and excretion. The anthocyanins had no effect in rabbits on the antioxidant capacity of plasma measured as Trolox equivalent antioxidant capacity and ferruc reducing ability of plasma.     

Extraction of polyphenols from processed black currant (Ribes nigrum L.) residues

The total phenol and anthocyanin contents of black currant pomace and black currant press residue (BPR) extracts, extracted with formic acid in methanol or with methanol/water/acetic acid, were studied. Anthocyanins and other phenols were identified by means of reversed phase HPLC, and differences between the two plant materials were monitored. In all BPR extracts, phenol levels, determined by the Folin-Ciocalteu method, were 8-9 times higher than in the pomace extracts. Acid hydrolysis liberated a much higher concentration of phenols from the pomace than from the black currant press residue. HPLC analysis revealed that delphinidin-3-O-glucoside, delphinidin-3-O-rutinoside, cyanidin-3-O-glucoside, and cyanidin-3-O-rutinoside were the major anthocyanins and constituted the main phenol class ( approximately 90%) in both types of black currant tissues tested. However, anthocyanins were present in considerably lower amounts in the pomace than in the BPR. In accordance with the total phenol content, the antioxidant activity determined by scavenging of 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation, the ABTS(*)(+) assay, showed that BPR extracts prepared by solvent extraction exhibited significantly higher (7-10 times) radical scavenging activity than the pomace extracts, and BPR anthocyanins contributed significantly (74 and 77%) to the observed high radical scavenging capacity of the corresponding extracts.     

Protease-assisted clarification of black currant juice: synergy with other clarifying agents and effects on the phenol content

Conventional clarification with gelatin and silica sol removes a considerable amount of antioxidant phenolics from berry juices. This study examined the clarification and haze-diminishing effects of alternative clarification strategies on black currant juice including centrifugation and addition of acidic protease and pectinolytic enzyme preparations and gallic acid. Centrifugation of freshly pressed juice (10,000 g for 15 min) resulted in a approximately 95% reduction of immediate turbidity and had a decreasing effect on haze development in the juice during cold storage without significantly compromising the total phenols levels. The extent of clarification and haze diminishment varied after individual treatments with five different acidic proteases, but one of the protease preparations, Enzeco, derived from Aspergillus niger, consistently tended to perform best. The individual and interactive effects on juice turbidity, total phenols, and total anthocyanin contents of clarification treatments involving the use of two selected acid proteases (Enzeco and Novozyme 89L), a pectinase (Pectinex BE 3-L), and gallic acid were evaluated in a full factorial 2(4) experimental design. Haze development during cold storage decreased when gallic acid or any of the enzyme preparations were employed individually, but negative interaction effects resulted when the pectinase was employed in combination with any of the proteases. After 28 storage days at 2 degrees C, the lowest levels of haze formation were achieved when the Enzeco protease preparation, added at 0.025 g/L, was added with 0.050 g/L of gallic acid and allowed to react in the juice for 90 min at 50 degrees C. The corresponding anthocyanin reduction was approximately 12% (compared to approximately 30% with gelatin silica sol treatment). The data support the hypothesis that phenol-protein interactions are involved in juice turbidity development during cold storage of berry juices and demonstrate that precentrifugation and protease-assisted clarification show promise as an alternative, phenolics-retaining clarification strategy in black currant juice processing.     

Stability and antioxidant activity of black currant anthocyanins in solution and encapsulated in glucan gel

The effects of ferric and ferrous ions, pH, and temperature on the stability and antioxidant activity of black currant anthocyanins (BCA) were studied, and the recovery of BCA from glucan gel [mixed linked (1-->3,1-->4)-beta-D-glucan] after using different encapsulating procedures was determined. The degradation of individual anthocyanins follows first-order kinetics and shows Arrhenius temperature dependence. The activation energies of individual anthocyanins, evaluated over the temperature range 60-100 degrees C, decrease with an increase in pH. While the antioxidant activity of BCA, measured by the ferric reducing antioxidant power assay, decreased with the degradation of anthocyanins, the completely degraded products still exhibited approximately 30% of the initial antioxidant activity. Ferric ions have a detrimental effect on the stability of BCA, especially for delphinidins. Freeze drying of encapsulated BCA gives approximately 20% higher recovery of individual anthocyanins than infrared drying.     

Antioxidant capacity of seed coat, dehulled bean, and whole black soybeans in relation to their distributions of total phenolics, phenolic acids, anthocyanins, and isoflavones

Black soybeans have been used as an excellent dietary source for disease prevention and health promotion in China for hundreds of years. However, information about the distribution of health-promoting phenolic compositions in different physical parts of black soybean and the contribution of phenolic compositions to overall antioxidant capacity is limited. To elucidate the distribution of phenolic composition and their contribution to antioxidant activities in black soybean, the total and individual phenolic profiles, and antioxidant capacities of seed coat, dehulled and whole black soybean were systematically investigated. The seed coat exhibited much higher total phenolic indexes and antioxidant activities than whole and dehulled black soybean. Dehulled black soybean possessed similar levels of total phenolic content, total flavonoid content, 2-diphenyl-1-picryhydrazyl (DPPH) radical scavenging activity, ferric reducing antioxidant power (FRAP), and oxygen radical absorbance capacity (ORAC) activities as compared to whole yellow soybean. Cyanidin-3-glucoside, petunidin-3-glucoside, and peonidin-3-glucoside were detected in the seed coat but not in dehulled black soybean and yellow soybean. Among benzoic acid detected, caffeic and chlorogenic acid were the predominant phenolic acids. Whole black soybean and dehulled black soybean exhibited similar isoflavone contents in 7- O-beta-glucosides and malonylglucosides of daidzein and genistein. The seed coat possessed significantly ( p < 0.05) lower 7- O-beta-glucosides and malonylglucosides of daidzein and genistein, acetylglycitin, and total isoflavones than whole and dehulled black soybean. The contribution of phenolics in the seed coat to the antioxidant activity of black soybean parts depends on the assay methods. When measured with the DPPH and FRAP methods, the seed coat contributed 90% of the total antioxidant capacity of black soybean. However, when measured with the ORAC method, the seed coat and dehulled portion contributed approximately equally the total antioxidant capacity of black soybeans. The information generated from this study on the distribution and content of their active components is useful for the effective use of black soybeans as an ingredient for promoting health.     

Influence of thermal treatment on black currant (Ribes nigrum L.) juice aroma

The influence of thermal treatment on black currant juice aroma was investigated in temperature and time ranges relevant for black currant juice concentration processes, namely, 45, 60, 75, and 90 degrees C. Forty-nine aroma compounds were quantified, and the thermal treatment resulted in concentration increases of most terpenes, aldehydes, furans, and phenols, whereas the concentration of esters slightly decreased. Higher temperatures and longer exposure times had larger effects on the aroma compounds. Odor triangle tests showed no sensory difference between pasteurized juice and juice heated at 60 degrees C, whereas juice heated at 90 degrees C differed significantly from pasteurized juice. It is concluded that a 90 degrees C thermal treatment of black currant juice, which is in the temperature range used for conventional evaporation of black currant juice, has an effect on the aroma and sensory properties.     

Fatty acid content and juice characteristics in black currant (Ribes nigrum L.) genotypes

The fatty acid compositions of seeds from 29 black currant genotypes were determined using a rapid small-scale procedure. There was interest in alpha-linolenic, stearidonic, and, especially, gamma-linolenic acid (GLA) contents, and most samples showed values between 11.1 and 18.7%, between 2.5 and 4.5%, and between 11.6 and 17.4%, respectively. However, six genotypes exhibited gamma-linolenic contents >18%, and values >20% were recorded in four of these genotypes. The fatty acid contents of the six genotypes were also analyzed by using a conventional procedure, and only slight differences in fatty acid composition were found between the two methods. Although GLA content was not strongly correlated with juice parameters, some genotypes had both high GLA contents and desirable juice characteristics. The results obtained provide evidence that it is possible to select for GLA contents without negatively affecting juice quality, and both aspects can be combined in a single cultivar, thereby increasing the added value of the whole fruit.     

Enzyme-assisted extraction of antioxidative phenols from black currant juice press residues (Ribes nigrum).

Enzymatic release of phenolic compounds from pomace remaining from black currant (Ribes nigrum) juice production was examined. Treatment with each of the commercial pectinolytic enzyme preparations Grindamyl pectinase, Macer8 FJ, Macer8 R, and Pectinex BE, as well as treatment with Novozym 89 protease, significantly increased plant cell wall breakdown of the pomace. Each of the tested enzyme preparations except Grindamyl pectinase also significantly enhanced the amount of phenols extracted from the pomace. Macer8 FJ and Macer8 R decreased the extraction yields of anthocyanins, whereas Pectinex BE and Novozym 89 protease showed no effect. A decrease in pomace particle sizes from 500-1000 microm to <125 microm increased the phenol yields 1.6-5 times. Black currant pomace devoid of seeds gave significantly higher yields of phenols than pomace with seeds and seedless wine pomace. Four selected black currant pomace extracts all exerted a pronounced antioxidant activity against human LDL oxidation in vitro when tested at equimolar phenol concentrations of 7.5-10 microM.     

Preparative-scale isolation of four anthocyanin components of black currant (Ribes nigrum L.) fruits

Four anthocyanin components of black currant, delphinidin 3-O-beta-rutinoside (D3R), cyanidin 3-O-beta-rutinoside (C3R), delphinidin 3-O-beta-glucoside (D3G), and cyanidin 3-O-beta-glucoside (C3G), were successfully isolated as crystalline forms on a preparative scale. In this process, selective hydrolysis of the glucosides (D3G and C3G) and rhamnosides (D3R and C3R) was achieved by treatment with beta-glucosidase and hesperidinase (alpha-rhamnosidase), respectively, to improve resolution of anthocyanin components. Especially, selective conversion of the rutinosides into glucosides made the amounts of D3G and C3G increase about 4- and 7-fold, respectively. D3R, C3R, D3G, and C3G were isolated from enzymatic hydrolysates of black currant anthocyanins through Amberlite XAD-7HP absorption followed by preparative HPLC separation, and their crystals were obtained as the flavylium chloride.     

Effects of mulching and cover cropping on soil microbial parameters in the organic growing of black currant

Abstract

Cover cropping and mulching to sustain and improve soil fertility and for weed control are common practices in organic growing systems. In this study, microbial parameters under different kinds of mulches and cover crops were analyzed in a field experiment with organically grown black currant (Ribes nigrum). The experiment comprised a combination of two mulches with bare soil as a control and two cover crops which were compared with bare soil, with and without an extra supply of organic fertilizer. Soil carbon (C) and nitrogen (N) as well as pH were unaffected by any of the treatments. The basal respiration rate was increased by mulching with wood chips throughout the four years of the experiment. During the last two years of the experiment, substrate induced respiration was also measured but was not found to be affected by any of the mulches. The potential ammonium (NH₄ ⁺) oxidation increased significantly after an initial supply of 200 kg N ha^‐1 as solid cattle manure. The increase was significantly lower under wood chips than in bare soil, although an extra 200 kg N ha^‐1 had been supplied under the wood chips. Furthermore, the black currant bushes suffered from a N deficiency in the wood chip treatment. The results showed that there was no substantial lasting build‐up of microbial biomass or organic matter content with wood chips because of lack of N, despite a large initial input of N and easily‐available C. Possible reasons for this deficiency are either increased denitrification under the wood chips or fungal translocation of N to the wood chip layer. Results from this experiment suggests that the evaluation of a few complementary biological soil parameters can be an important tool when developing sustainable growing systems and for indicating environmental stress.     

Antioxidant capacity and phenolic content of sweet rowanberries

Sweet rowanberry cultivars adapted to northern climates have been developed from rowanberries (Sorbus aucuparia L.) and hybrids of rowanberry with Malus, Pyrus, Aronia, or Mespilus. The rowanberries studied here (cvs. Burka, Dessertnaja, Eliit, Granatnaja, Kubovaja, Rosina, Rubinovaja, Titan, and Zholtaja) have high antioxidant and phenolic contents. The phenolic content varied between 550 and 1014 mg/100 g of fresh weight in sweet rowanberries, whereas 846 and 717 mg were found in the well-characterized bilberry and lingonberry, respectively. Anthocyanins (6-80 mg) were mainly found from berries of hybrid cultivars. Of the other phenolics, chlorogenic (29-160 mg) and neochlorogenic (34-104 mg) acids constituted the major fraction in all rowanberries, the concentrations almost equaling those present in coffee. Antioxidant capacities of rowanberries were high, as measured with FRAP (61-105 micromol of Fe2+/g) and DPPH (21.3-9.7 g/g DPPH) methods. Principal component analysis was able to separate the cultivars of different origin into clusters on the basis of their phenolic profiles.     

Effect of clarification techniques and rat intestinal extract incubation on phenolic composition and antioxidant activity of black currant juice

This study examined the phenolic composition and the antioxidant potencies of black currant juices that had been experimentally clarified with acidic proteases and pectinases to retain the phenolics and which had been subjected to rat intestinal mucosa extract incubation to mimic gut cell mediated biotransformation of phenolics. When compared at equimolar levels of 2.5 microM gallic acid equivalents, the black currant juice samples prolonged the induction time of human low-density lipoprotein oxidation in vitro by 2.6-3.6 times, and the order of antioxidant potency of differently clarified black currant juices was centrifuged juice > gelatin silica sol clarified juice > enzymatically clarified juice approximately raw juice. No immediate relationship between the, almost similar, phenolic profiles of the juice samples and their relative antioxidant activities could be established. Incubation of juices with a rat small intestine cell extract for 19 h promoted significant decreases in the contents of the anthocyanin 3-O-beta-glucosides (cyanidin 3-O-beta-glucoside and delphinidin 3-O-beta-glucoside), but did not affect the anthocyanin 3-O-beta-rutinosides (cyanidin 3-O-beta-rutinoside and delphinidin 3-O-beta-rutinoside) of the black currant juice. Black currant juice samples subjected to such intestinal cell extract incubation had approximately 30% decreased antioxidant capacity. Incubation of juices with the rat small intestine cell extracts at neutral pH appeared to decrease the levels of delphinidin glucosides more than the levels of cyanidin glucosides. The results provide an explanation for the predominant detection of anthocyanin rutinosides, and not anthocyanin glucosides, in plasma and urine in in vivo studies and provide important clues to better understand the complex mechanisms affecting dietary phenols in the gut.     

Effectiveness of biodiversity indicators varies with extent, grain, and region

The use of indicator taxa for conservation planning is common, despite inconsistent evidence regarding their effectiveness. These inconsistencies may be the result of differences among species and taxonomic groups studied, geographic location, or scale of analysis. The scale of analysis can be defined by grain and extent, which are often confounded. Grain is the size of each observational unit and extent is the size of the entire study area. Using species occurrence records compiled by NatureServe from survey data, range maps, and expert opinion, we examined correlations in species richness between each of seven taxa (amphibians, birds, butterflies, freshwater fish, mammals, freshwater mussels, and reptiles) and total richness of the remaining six taxa at varying grains and extents in two regions of the US (Mid-Atlantic and Pacific Northwest). We examined four different spatial units of interest: hexagon (∼649 km²), subecoregion (3800-34,000 km²), ecoregion (8300-79,000 km²), and geographic region (315,000-426,000 km²). We analyzed the correlations with varying extent of analysis (grain held constant at the hexagon) and varying grain (extent held constant at the region). The strength of correlation among taxa was context dependent, varying widely with grain, extent, region, and taxon. This suggests that (1) taxon, grain, extent, and study location explain, in part, inconsistent results of previous studies; (2) planning based on indicator relationships developed at other grains or extents should be undertaken cautiously; and (3) planning based on indicator relationships developed in other geographic locations is risky, even if planning occurs at an equivalent grain and extent.