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.     

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.     

Enzyme-assisted processing increases antimicrobial and antioxidant activity of bilberry

The effects of nine cell wall-degrading enzymes on the antimicrobial and antioxidant activities of bilberry were studied. Antimicrobial activity was measured using the human pathogens Salmonella enterica sv. Typhimurium and Staphylococcus aureus as test strains. Enzyme treatments liberated phenolics from the cell wall matrix, which clearly increased the antimicrobial activity of berry juices, press cakes, and berry mashes on the basis of plate counts. Antibacterial effects were stronger against Salmonella than against Staphylococcus bacteria. In general, the increase in activity measured as colony-forming units per milliliter was 3-5 logarithmic units against Salmonella and 1-2 units against Staphylococcus bacteria. Increase in antimicrobial activity was observed only in acidic conditions, which is also the natural environment in various berry products, such as juices. The activity profile of the pectinase preparation affected the chemistry of the phenolics due to the presence of deglycosylating activities in some preparations. The difference in phenolic profiles was reflected in the antimicrobial effects. Bilberry mashes treated with Pectinex Ultra SP-L, Pectinex 3 XL, and Pectinex BE XXL were most efficient against Salmonella bacteria, whereas mashes treated with Pectinex Smash, Pectinex BE 3-L, and Biopectinase CCM showed the strongest antimicrobial activity against Staphylococcus bacteria. Due to the liberation of phenolics from the cell wall matrix the antioxidant activity measured as radical scavenging activity was also increased on average about 30% by the enzymatic treatments. The highest increase in phenolic compounds was about 40%. Highest increases in anthocyanins and in antioxidant activity were observed in berry mash treated with Pectinex Smash XXL enzyme, and the lowest increase was observed after treatment with Pectinex BE 3-L. Enzyme-assisted processing is traditionally used to improve berry and fruit juice yields. However, enzymatic treatments also have an impact on the functional properties of the products. The increased liberation of phenolics from the cell wall matrix can prolong the shelf life of berry products by limiting the growth of contaminants during processing or storage. The increased amount of phenolic compounds may also have a positive effect on gut well-being.     

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.     

Efficiency of enzymatic and other alternative clarification and fining treatments on turbidity and haze in cherry juice

Several alternative strategies were examined for improving conventional juice fining procedures for cherry juice clarification and fining in laboratory-scale experiments: Centrifugation of freshly pressed juice from 1000g to 35,000g induced decreased turbidity according to a steep, negative power function. Individual and interactive effects on turbidity and haze formation in precentrifuged and uncentrifuged cherry juice of treatments with pectinase, acid protease, bromelain, gallic acid, and gelatin-silica sol were investigated in a factorial experimental design with 32 different parameter combinations. Gelatin-silica sol consistently had the best effect on juice clarity. Centrifugation of cherry juice (10,000g for 15 min) prior to clarification treatment significantly improved juice clarity and diminished the rate of haze formation during cold storage of juice. Both treatment of precentrifuged cherry juice with Novozym 89L protease and co-addition of pectinase and gallic acid improved cherry juice clarity and diminished haze levels. None of the alternative treatments produced the unwieldy colloids notorious to gelatin-silica sol treatment. The data suggest that several alternative clarification strategies deserve further consideration in large-scale cherry juice processing. Precentrifugation of juice before clarification and fining is immediately recommended.     

Effect of black currant anthocyanins on the activation of endothelial nitric oxide synthase (eNOS) in vitro in human endothelial cells

Polyphenols are known to induce vasodilatory function via activation of the redox-sensitive phosphatidylinositol-3 (PI3)/protein kinase B (Akt) pathway. Black currant fruits have appreciable amounts of polyphenolic compounds including cyanidin-3-O-glucoside, cyanidin-3-O-rutinoside, delphinidin-3-O-glucoside, and delphinidin-3-O-rutinoside. It was hypothesized that black currant fruit extracts would cause activation of endothelial nitric oxide synthase (eNOS) through activation of redox-sensitive PI3 kinase/Akt signaling pathway. To test this hypothesis, human umbilical vein endothelial cells (HUVECs) were treated with different concentrations/times of black currant juice concentrates (Ben Gairn and Ben Hope) and the activation of Akt and eNOS was measured using immunoblotting. Vitamin C is also known to activate Akt and eNOS in in vitro models, and black currants are rich in vitamin C. Therefore, the effect of black currant extracts with and without coexisting vitamin C was investigated, using SPE columns to eliminate vitamin C content. The individual (and combined) effects of the major anthocyanins present in black currant juice samples with and without vitamin C were investigated and compared to the effects of the whole extract. Black currant juice samples (1 μL/mL) significantly increased the phosphorylation of Akt (p-Akt) and eNOS (p-eNOS) (P < 0.05). Activation of Akt and eNOS was abolished by incubation with wortmannin, a PI3K inhibitor, supporting the involvement of PI3K/Akt. Vitamin C alone significantly increased the p-Akt and p-eNOS (P < 0.05); however, removal of vitamin C from black currant did not significantly affect p-Akt and p-eNOS compared to black currant with vitamin C. Assessment of individual anthocyanins also showed significant effects on p-Akt and p-eNOS. In summary, in the present study data suggested that black currant concentrates, Ben Gairn and Ben Hope, activated eNOS via Akt/PI3 kinase pathway in vitro in HUVECs and that the effect was not dependent on vitamin C.     

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.     

Anthocyanins in berries of ribes including gooseberry cultivars with a high content of acylated pigments

Consumption of berries from various sources including the genus Ribes has been associated with diverse potential health benefits. The 14 examined cultivars of European gooseberry (R. grossularia L.) contained in various proportions the 3-glucoside (3), 3-rutinoside (4), 3-xyloside (7), 3-O-beta-(6' '-E-caffeoylglucopyranoside) (8), and 3-O-beta-(6' '-E-p-coumaroylglucopyranoside) (10) of cyanidin and the 3-rutinoside (6) and 3-glucoside of peonidin (5). Pigments 3, 4, delphinidin 3-rutinoside (2), delphinidin 3-glucoside (1), and minor amounts of 6, 7, and 10 were found in red flowering currant (R. sanguineum Pursh). Golden currant (R. aureum Pursh) contained 3, 4, and trace amounts of 1, 6, and 7, while alpine currant (R. alpinum L.) contained 3, 4, and trace amounts of 10. The major anthocyanins in two cultivars of jostaberries (R. x nidigrolaria Bauer), 1-4, 8, and 10, reflected that this hybrid contained the major anthocyanins of both parents, black currant and gooseberry. This is the first complete identification of 8 and the ring size of the sugar of 10. Pigment 9 was tentatively identified as cyanidin 3-(6' '-Z-p-coumaroylglucoside). This new pigment occurred in minor amounts (<2%) in all R. grosssularia and R. x nidigrolaria cultivars. No commercially available berries have been reported to contain such high proportions of aromatic acylated anthocyanins as found in the gooseberry cultivars "Sams?", "Hinnom?ki Red", "Taastrup", "Lofthus", and "Glendal", which are in this context the most obvious candidates for consumption, colorant, and breeding programs.     

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.     

Content of the flavonols quercetin, myricetin, and kaempferol in 25 edible berries

The amounts of quercetin, myricetin, and kaempferol aglycons in 25 edible berries were analyzed by an optimized RP-HPLC method with UV detection and identified with diode array and electrospray ionization mass spectrometry detection. Sixteen species of cultivated berries and nine species of wild berries were collected in Finland in 1997. Quercetin was found in all berries, the contents being highest in bog whortleberry (158 mg/kg, fresh weight), lingonberry (74 and 146 mg/kg), cranberry (83 and 121 mg/kg), chokeberry (89 mg/kg), sweet rowan (85 mg/kg), rowanberry (63 mg/kg), sea buckthorn berry (62 mg/kg), and crowberry (53 and 56 mg/kg). Amounts between 14 and 142 mg/kg of myricetin were detected in cranberry, black currant, crowberry, bog whortleberry, blueberries, and bilberry. Kaempferol was detected only in gooseberries (16 and 19 mg/kg) and strawberries (5 and 8 mg/kg). Total contents of these flavonols (100-263 mg/kg) in cranberry, bog whortleberry, lingonberry, black currant, and crowberry were higher than those in the commonly consumed fruits or vegetables, except for onion, kale, and broccoli.     

Ultrasonic extraction of phenols from olive mill wastewater: comparison with conventional methods

Recovery of phenols from olive mill wastewater (OMWW) was studied, comparing five sample preparation methods: filtration, solid-phase (SPE), liquid-liquid (LLE) and ultrasonic (US)-assisted extraction of liquid and solid (freeze-dried) OMWW. Results showed that ultrasonication is a good alternative to conventional solvent extractions, providing higher recoveries at both levels of individual and total phenol yields. Sonication of liquid OMWW in organic solvent was more efficient vs its nonassisted counterpart (agitation), but did not provide a representative phenol chromatogram due to ethyl acetate use. By contrast, the US-assisted extraction of freeze-dried OMWW (3 × 20 min) in 100% methanol (1.5 g/25 mL, w/v) offered the highest qualitative-quantitative phenol yields without any US-induced alterations. Moreover, freeze-drying is an excellent preservation of initial liquid OMWW, holding a great potential for delayed analysis. This study is also the first report that Slovenian OMWW may be utilized as a valuable source of phenols, especially hydroxytyrosol and tyrosol.     

Black currant response to foliar fertilizers – modeling of varietal growth dynamics

Abstract

Black currant cultivation is in continuous expansion, with importance in human nutrition and a high degree of suitability for industrial processing. The nutritional, organoleptic, and pharmaceutical qualities of its fruits represent a great source of natural antioxidants, vitamins, and phenols. We have tested the specific reaction of three black currant varieties to foliar fertilizers in order to obtain two growth models. Each variety have a specific response to fertilizers. Application of Kombimax lead to the highest increase during the vegetation period and 72?cm length of branches at the end of vegetation period. Ascovigor and Mikromix produces 3–4?cm increases in branches length compared to control. N, K2O, and CaO from fertilizer recipe act synergically for the final development of branches. The growth model based on branches length at 14?days add 0.97?cm for each existent cm. The complex model of growth add 2.44?cm for each mg of CaO existent in fertilizer recipe. Linear regressions show a growth potential of 0.97?cm/1?cm of branches at 14?days.     

Biophenols in table olives

Unprocessed olives are well-known sources of phenolic antioxidants with important biological properties. Processing methods to prepare table olives may cause a reduction of valuable phenols and may deprive the food of precious biological functions. The present work was undertaken to evaluate table olives produced in Greece as sources of biophenols. Commercially available olives were analyzed for their total phenol content by using the Folin-Ciocalteu reagent and for individual phenols by RP-HPLC. Samples were Spanish-style green olives in brine, Greek-style naturally black olives in brine, and Kalamata olives in brine. Most of the types of olives analyzed were found to be good sources of phenols. Hydroxytyrosol, tyrosol, and luteolin were the prevailing phenols in almost all of the samples examined. High levels of hydroxytyrosol were determined mainly in Kalamata olives and Spanish-style green olives, cultivar Chalkidiki (250-760 mg/kg).     

Antioxidant compounds and antioxidant activity in "early potatoes"

The antioxidant content and the antioxidant capacity of both hydrophilic and lipophilic antioxidant extracts from four "early potato" cultivars, grown in two different locations (Racale and Monteroni), were examined. There was a considerable variation in carotenoid content and weak differences in the ascorbic acid concentration of the examined cultivars of "early potato" and between the harvested locations. An increase in both methanol/water (8:2 v/v) and phosphate buffer soluble (PBS) free phenols (70%) and bound phenols (28%) in the extracts from the cultivars grown at Racale site was found and discussed. Examination of individual phenols revealed that chlorogenic acid and catechin were the major phenols present in potato tuber extracts; a moderate amount of caffeic acid and ferulic acid was also detected. The total equivalent antioxidant capacity (TEAC) was higher in the Racale extracts and a highly positive linear relationship ( R (2) = 0.8193) between TEAC values and total phenolic content was observed. The oxyradical scavenging capacity (TOSC) of methanol/water and PBS extracts of peel and whole potatoes against the reactive oxygen species (ROS) peroxyl radicals, peroxynitrite, and hydroxyl radicals was also analyzed. A highly significant linear correlation ( R (2) = 0.9613) between total antioxidant capacity (as a sum of peroxyl radicals + peroxynitrite) and total phenol content of methanol/water extracts was established. Moreover, proliferation of human mammalian cancer (MCF-7) cells was significantly inhibited in a dose-dependent manner after exposure to potato extracts. These data can be useful for "early potato" tuber characterization and suggest that the "early potato" has a potential as a dietary source of antioxidants.     

Characterization of phenolic profiles of Northern European berries by capillary electrophoresis and determination of their antioxidant activity

Berries are known to contain phenolic substances (i.e., flavonoids and phenolic acids), which comprise two large and heterogeneous groups of biologically active nonnutrients. This investigation evaluated the content and profile of the phenolic compounds present in six different berries found in Northern Europe. The latter included bilberry (Vaccinium myrtillus), cowberry (Vaccinium vitis-idaea), cranberry (Vaccinium oxycoccus), strawberry (Fragaria ananassa), black currant (Ribes nigrum), and red currant (Ribes rubrum). The study was focused on two areas. The first involved the extraction and analysis of berries for total phenolic content and determination of their antioxidant activity. The total phenolic level of berries was correlated with their antioxidant activity. Second, the berry extracts were analyzed by capillary electrophoresis to determine the content and profile of selected bioactive compounds. The analytes of interest included trans-resveratrol, cinnamic acid, ferulic acid, p-coumaric acid, quercetin, and morin.     

High-performance liquid chromatography analysis of black currant (Ribes nigrum L.) fruit phenolics grown either conventionally or organically

Black currants (Ribes nigrum L.) contain a diverse range of phenolics and possess a high antioxidant activity, which makes them an interesting target for the functional food industry. In this study, phenolic profiles of organically and conventionally grown black currant fruits, collected from commercial farms within a climatically similar area, were compared. Compounds were identified using UV/vis and mass spectroscopy techniques and quantified with high-performance liquid chromatography equipped with UV/vis detection. Several different conjugates of hydroxycinnamic acids, flavonols, and anthocyanins were quantified. Statistically significant differences between farms were found for almost all compounds. Differences between the highest and the lowest measured values of major phenolic compounds of different phenolic classes ranged from 24 to 77%. Principal component analysis quite effectively separated farms from each other but did not cluster them according to cultivation technique. Thus, it was concluded that the biochemical quality of organically grown black currant fruits does not differ from those grown conventionally.     

黑豆丹贝发酵前后香气、酚类物质及抗氧化活性分析

为了丰富丹贝品种并提高黑豆食品加工利用率,本试验以少孢根霉(Rhizopus oligtosporus)发酵制备黑豆丹贝,测定其基本营养成分和香气成分,并分析不同发酵时间黑豆丹贝中麦角固醇、氨基酸态氮、10 kDa以下肽、总酚、总黄酮、原花青素含量及蛋白质水解度抗氧化活性的变化.结果表明,与未发酵样品相比,发酵后黑豆丹...     

Plant phenolics affect oxidation of tryptophan

The effect of berry phenolics such as anthocyanins, ellagitannins, and proanthocyanidins from raspberry (Rubus idaeus), black currant (Ribes nigrum), and cranberry (Vaccinium oxycoccus) and byproducts of deoiling processes rich in phenolics such as rapeseed (Brassica rapa L.), camelina (Camelina sativa), and soy (Glycine max L.) as well as scots pine bark (Pinus sylvestris) was investigated in an H2O2-oxidized tryptophan (Trp) solution. The oxidation of Trp was analyzed with high-performance liquid chromatography using both fluorescence and diode array detection of Trp and its oxidation products. Mechanisms of antioxidative action of the phenolic compounds toward the oxidation of Trp were different as the pattern of Trp oxidation products varied with different phenolic compounds. The antioxidant protection toward oxidation of Trp was best provided with pine bark phenolics, black currant anthocyanins, and camelina meal phenolics as well as cranberry proanthocyanidins.     

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.     

Halogenated phenols in water at forty Canadian potable water treatment facilities

Samples of raw and treated water were collected once in each of 3 seasons at 40 potable water treatment plants across Canada and were analyzed for phenol and 33 halogenated phenolic compounds including chlorophenols, bromophenols, bromochlorophenols, and chloroguaiacols. Eighteen of the compounds were not found at any treatment plant; phenol and each of the remaining halogenated phenols were found in at least 1 sample. Pentachlorophenol was the only halogenated phenolic compound found in more than 20% of the raw water samples in the fall and winter samples at levels up to 53 ng/L with mean values of 1.9 and 2.8 ng/L, respectively. No halogenated phenols were detected in raw water summer samples. The halogenated phenols found most frequently in treated water samples were 4-chloro-, 2,4-dichloro-, 2,4,6-trichloro-, and bromodichlorophenols. Mean values were less than 15 ng/L and maximum values seldom exceeded 100 ng/L. Most of the positive values for the treated water samples were found at 8 of the 40 treatment plants but no correlations could be found between halogenated phenol levels and raw water type, treatment process, or chemical dosages.