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Polyphenols and antioxidant capacity of seed coat and cotyledon from Brazilian and Peruvian bean cultivars (Phaseolus vulgaris L.).

Seed coats and cotyledons from 25 Brazilian and 3 Peruvian bean cultivars were investigated in relation to their phenolic profiles and antioxidant capacity. Condensed tannins, anthocyanins, and flavonols such as kaempferol and quercetin glycosides were mostly found in seed coats. Cotyledons were rich in phenolic acids, such as ferulic, sinapic, chlorogenic, and other hydroxycinnamic acids. In general, the seed coat color pattern and the type of cultivar showed an important influence on the variability of phenolic profiles and levels, respectively. Total phenolics and antioxidant capacity assessed by the DPPH method were higher in seed coats than in cotyledons. The antioxidant capacity had a significant correlation with condensed tannins for all samples and with total anthocyanins in black and red seed coats, whereas in cotyledons, it was more related to the total phenolic content.

Anthocyanins↗

Interactions of different phenolic acids and flavonoids with soy proteins.

Soy glycinin (SG) and soy trypsin inhibitor (STI) were derivatized by chlorogenic- and caffeic acid (cinnamic acids, C(6)-C(3) structure), and by gallic acid representing hydroxybenzoic acids (C(6)-C(1) structure). Further, the flavonoids, flavone, apigenin, kaempferol, quercetin and myricetin (C(6)-C(3)-C(6) structure) were also caused to react with soy proteins to estimate the influence of the number and the position of hydroxy substituents. The derivatization caused a reduction of lysine, cysteine and tryptophan residues in the soy proteins. The isoelectric points of the derivatives were shifted to lower pH values and formation of high molecular fractions was documented. The derivatives were characterized in terms of their solubility at different pH-values to document the influence on the functional properties. The structural changes induced were studied using circular dichroism (CD), differential scanning calorimetry (DSC), intrinsic fluorescence, and binding of anilinonaphthalenesulfonic acid. The influence of derivatization on the in-vitro digestibility with trypsin, chymotrypsin, pepsin and pancreatin was also assessed. The effect on the trypsin inhibitor activity of all the resulting STI derivatives was studied, the latter being reduced.

Anilino Naphthalenesulfonates↗

Protein binding in deactivation of ferrylmyoglobin by chlorogenate and ascorbate.

Kinetics of reduction of iron(IV) in ferrylmyoglobin by chlorogenate in neutral or moderately acidic aqueous solutions (0.16 M NaCl) to yield metmyoglobin was studied using stopped flow absorption spectroscopy. The reaction occurs by direct bimolecular electron transfer with (2.7 +/- 0.3) x 10(3) M(-)(1).s(-)(1) at 25.0 degrees C (DeltaH( )(#) = 59 +/- 6 kJ.mol(-)(1), DeltaS(#) = 15 +/- 22 J. mol(-)(1).K(-)(1)) for protonated ferrylmyoglobin (pK(a) = 4.95) and with 216 +/- 50 M(-)(1).s(-)(1) (DeltaH( )(#) = 73 +/- 8 kJ. mol(-)(1), DeltaS( )(#) = 41 +/- 30 J.mol(-)(1).K(-)(1)) for nonprotonated ferrylmyoglobin in parallel with reduction of a chlorogenate/ferrylmyoglobin complex by a second chlorogenate molecule with (8.6 +/- 1.1) x 10(2) M(-)(1).s(-)(1) (DeltaH( )(#) = 74 +/- 8 kJ.mol(-)(1), DeltaS( )(#) = 59 +/- 28 J.mol(-)(1).K(-)(1)) for protonated ferrylmyoglobin and with 61 +/- 9 M(-)(1).s(-)(1) (DeltaH( )(#) = 82 +/- 12 kJ.mol(-)(1), DeltaS( )(#) = 63 +/- 41 J. mol(-)(1).K(-)(1)) for nonprotonated ferrylmyoglobin. Previously published data on ascorbate reduction of ferrylmyoglobin are reevaluated according to a similar mechanism. For both protonated and nonprotonated ferrylmyoglobin the binding constant of chlorogenate is approximately 300 M(-)(1), and the modulation of ferrylmyoglobin as an oxidant by chlorogenate (or ascorbate) leads to a novel antioxidant interaction for reduction of ferrylmyoglobin by ascorbate in mixtures with chlorogenate.

Ascorbic Acid↗

The active principles of plant extracts with antithyrotropic activity: oxidation products of derivatives of 3,4-dihydroxycinnamic acid.

We have recently reported that freeze-dried extracts (FDE) of certain plants form high molecular weight adducts with bovine TSH (bTSH), preventing it from binding to and stimulating adenylate cyclase in human thyroid membranes. We have now studied 34 pure compounds identical or structurally related to compounds present in FDE from Lycopus or Lithospermum, 2 of the 3 species of active plants studied previously. In studies conducted at 4 C in 20 mM Tris-HCl-0.5% BSA buffer, pH 7.45, eight 3,4-dihydroxylated compounds, all structurally related to cinnamic acid, inhibited the binding of [125I] bTSH to human thyroid membranes. Of these, 4 (caffeic, rosmarinic, chlorogenic, and ellagic acids) are present in the plants, and 4 (3,4-dihydroxyphenylacetic acid, deoxyepinephrine, adenochrome, and nordihydroguaretic acid) are structurally related thereto. These compounds were inactive when tested directly but became active when allowed to undergo auto-oxidation. With all 8 compounds, half-maximum inhibition of [125I]bTSH binding required quantities of oxidized product equivalent to 20-80 micrograms/ml (60-195 microM) of the original compound. Half-maximum inhibitory concentrations of oxidized caffeic and ellagic acids were increased 2- to 4-fold when experiments were performed at 37 C in medium containing 50 mM NaCl. Preincubation of membranes with active oxidation products in concentrations up to 100 micrograms/ml, followed by washing, had no effect on the subsequent binding of [125I]bTSH. As has been shown in the case of FDE, when [125I]bTSH was preincubated with oxidation products of caffeic and ellagic acids and was then chromatographed on Sephadex G-100, its elution pattern was advanced from an apparent mol wt of 30,000 to the void volume, and [125I]bTSH in the early eluting fractions displayed greatly reduced binding to thyroid membrane preparations. Addition of a large excess of unlabeled bTSH during preincubation prevented the shift in the elution pattern of [125I]bTSH produced by these oxidation products. To ascertain whether FDE and active compounds interact with the protein or carbohydrate moieties of bTSH, studies of their effects on the binding and chromatographic behavior of 125I-deglycosylated-bTSH (dg-bTSH) were also performed. Effects were similar to those observed for intact bTSH, suggesting that they do not interact with the carbohydrate moiety of TSH. Preincubation of both bTSH and dg-bTSH with either active FDE or oxidation products of caffeic or rosmarinic acid also greatly decreased their activity in the McKenzie mouse assay.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

From plums to prunes: influence of drying parameters on polyphenols and antioxidant activity.

Prunes, which are industrially obtained by dehydrating fresh plums at 85-90 degrees C for 18 h, contain higher levels of phenolic compounds than most other fruits. Prune phenolics have shown beneficial effects on human health. Reports are available in the literature on ascorbic acid, phenol composition, and antioxidant activity of fresh plums and prunes, but there is a lack of publications on the influence of drying parameters on the phenolic compounds and antioxidant activity. A study was carried out on two plum cultivars using two sets of air-drying temperatures: (i) air temperature at 85 degrees C until 50% of prune moisture level and then the temperature was lowered to 70 degrees C; (ii) air temperature at 60 degrees C. Whole fresh and dried fruits were assessed for phenolics (catechins, hydroxycinnamic acids, anthocyanins, and flavonols), ascorbic acid, and antioxidant activity (all parameters were calculated on a dry matter basis). Analysis of data shows that chlorogenic and neochlorogenic acid changes were affected by both process parameters and cultivar. Drying destroyed anthocyanins, and there was a significant decrease in flavonols. Ascorbic acid was drastically reduced in relation to process temperature. The most striking result was that drying at 85 degrees C doubled antioxidant activity in both cultivars, while contradictory results were found for 60 degrees C processed plums.

Antioxidants↗

Characterization and quantitation of phenolic compounds in new apricot (Prunus armeniaca L.) varieties.

Thirty-seven apricot varieties, including four new releases (Rojo Pasión, Murciana, Selene, and Dorada) obtained from different crosses between apricot varieties and three traditional Spanish cultivars (Currot, Mauricio, and Búlida), were separated according to flesh color into four groups: white, yellow, light orange, and orange (mean hue angles in flesh were 88.1, 85.0, 77.6, and 72.4, respectively). Four phenolic compound groups, procyanidins, hydroxycinnamic acid derivatives, flavonols, and anthocyanins, were identified by HPLC-MS/MS and individually quantified using HPLC-DAD. Chlorogenic and neochlorogenic acids, procyanidins B1, B2, and B4, and some procyanidin trimers, quercetin 3-rutinoside, kaempferol 3-rhamnosyl-hexoside and quercetin 3-acetyl-hexoside, cyanidin 3-rutinoside, and 3-glucoside, were detected and quantified in the skin and flesh of the different cultivars. The total phenolics content, quantified as the addition of the individual compounds quantified by HPLC, ranged between 32.6 and 160.0 mg 100 g(-1) of edible tissue. No correlation between the flesh color and the phenolic content of the different cultivars was observed.

Anthocyanins↗

Leaf extracts of some Cordia species: analgesic and anti-inflammatory activities as well as their chromatographic analysis.

Leaf preparations of several species of Cordia are used in traditional medicine as remedies for osteoarticular diseases. The analgesic, anti-inflammatory and anti-arthritic activities in the rat of different extracts of C. francisci, C. martinicensis, C. myxa, C. serratifolia and C. ulmifolia leaves were studied. The results obtained showed that the petroleum ether and alcoholic extracts especially of C. francisci, C. myxa and C. serratifolia leaves have a significant analgesic, anti-inflammatory and anti-arthritic activity in the rat. The flavonoids and phenolic derivative content of the five species of genus Cordia leaves was investigated by TLC and determined by reversed-phase HPLC with an acetonitrile/water/acetic acid buffer solvent gradient. UV detection was carried out at 255 and 280 nm. Four flavonoid glycosides, robinin, rutin, datiscoside and hesperidin, one flavonoid aglycone, dihydrorobinetin, two phenolic derivatives, chlorogenic and caffeic acid, were evidenced and determined.

Analgesics↗

Sugars and Organic Acids of Vitis vinifera.

Glucose, fructose, galactose, sucrose, maltose, melibiose, raffinose, and stachyose were identified in the leaves, bark, roots, and berries of Vitis vinifera L. var. Thompson Seedless. In addition to these sugars, verbascose and manninotriose were found in the leaves and bark.Malic, tartaric, citric, isocitric, ascorbic, cis-aconitic, oxalic, glycolic, glyoxylic, succinic, lactic, glutaric, fumaric, pyrrolidone carboxylic, alpha-ketoglutaric, pyruvic, oxaloacetic, galacturonic, glucuronic, shikimic, quinic, chlorogenic, and caffeic acids were identified in the leaves, bark, roots, and berries.Glucose, fructose, sucrose, malate, tartrate, and citrate were determined quantitatively in the leaf, petiole, xylem, bark, tendril, bud, puduncle pedicel, berry, lateral roots, and main roots at 4 separate physiological stages of growth. In addition, changes in the concentrations of fructose, glucose, malate, and tartrate in leaves were measured during a 36-day period starting from budburst.

Journal Article↗

Antioxidant activity of the microalga Chlorella vulgaris cultered on special conditions.

The chemical composition of Chlorella vulgaris indicates that it has a high nutritional value to a wide range of essential nutrients, such as vitamins, minerals and proteins. Moreover, it contains other compounds such as n-3 and n-6 polynsaturated fatty acids, provitamins and phenolic compounds. In addition, this alga can be produced in large-scale systems. The objective of the present study was to evaluate the antioxidant capacity of a Chlorella cultured on three differents temperatures (15 degrees C, 20 degrees C and 30 degrees C) in 3 Klux. Chlorella cultured samples were submitted to sequential extration using as solvents: ether, methanol and water. The antioxidant activity in the extracts was measured by b-carotene/linoleic acid system, at 50 degrees C and absorbances reading at 470 nm. One control with BHT, 100 ppm was used in this determination. The total phenolic compounds was determined with Folin-Ciocalteu reagent using the spectrophotometric measured at 780 nm with catechin as standard. The phenolic acid analysis were carried out using gas chromatograph equipped with a capillary column and flame ionization detector. Non conjugated and total phenolic acids were identified on the basis of the relative retention time of their derivatives compared with the standard phenolic acids. The methanolic extract from Chlorella cultured at 30 degrees C showed higher antioxidant activity (85%) quite similar of BHT (86%). By the Rancimat test (lipidic medium) two fractions from methanolic extracts showed too higher antioxidant activity with induction times > 37.50 h at 60 degrees C and 11.5 h at 100 degrees C. The total phenolic compounds were 24.95 mg in 1 g of dry alga matter from methanolic extract and five phenolic acids were identified. The phenolic compounds salicylic, trans cinnamic, synaptic, chlorogenic, chimic and caffeic acids found in the methanolic Chlorella extract may be responsible for its higher antioxidant activity.

Antioxidants↗

Use of the 1H nuclear magnetic resonance spectra signals from polyphenols and acids for chemometric characterization of cider apple juices.

The low field region (5.8-9.0 ppm) corresponding to aromatic protons and the region 1.8-3.0 ppm of the (1)H NMR spectra were used for characterization and chemometric differentiation of 52 apple juices obtained from six cider apple varieties. The data set consisted of 14 integrated areas corresponding to resonances from acids and phenolic compounds. Multivariate procedures based on hierarchical cluster and discriminant analysis were performed on selected signals of the spectra to determine whether it was possible to distinguish the different juices. Cluster analysis was able to satisfactorily classify the six apple varieties. Discriminant analysis, by means of stepwise procedure for variables selection and leave-one-out for cross-validation, was applied to 40 samples from the year 2001, obtaining recognition and prediction abilities of 100%. The most discriminant variables corresponded to poliphenols, (-)-epicatechin, phloridzin-phloretin, and p-coumaric, chlorogenic, and malic acids. The classification model was applied to 12 samples from apples harvested in the years 2002 and 2003, and the prediction ability was 91.7%.

Beverages↗

Purification and characterization of a new cationic peroxidase from fresh flowers of Cynara scolymus L.

A basic heme peroxidase isoenzyme (AKPC) has been purified to homogeneity from artichoke flowers (Cynara scolymus L.). The enzyme was shown to be a monomeric glycoprotein, M(r)=42300+/-1000, (mean+/-S.D.) with an isoelectric point >9. The native enzyme exhibits a typical peroxidase ultraviolet-visible spectrum with a Soret peak at 404 nm (epsilon=137,000+/-3000 M(-1) cm(-1)) and a Reinheitzahl (Rz) value (A(404nm)/A(280nm)) of 3.8+/-0.2. The ultraviolet-visible absorption spectra of compounds I, II and III were typical of class III plant peroxidases but unlike horseradish peroxidase isoenzyme C, compound I was unstable. Resonance Raman and UV-Vis spectra of the ferric form show that between pH 5.0 and 7.0 the protein is mainly 6 coordinate high spin with a water molecule as the sixth ligand. The substrate-specificity of AKPC is characteristic of class III (guaiacol-type) peroxidases with chlorogenic and caffeic acids, that are abundant in artichoke flowers, as particularly good substrates at pH 4.5. Ferric AKPC reacts with hydrogen peroxide to yield compound I with a second-order rate constant (k(+1)) of 7.4 x 10(5) M(-1) s(-1) which is significantly slower than that reported for most other class III peroxidases. The reaction of ferric and ferrous AKPC with nitric oxide showed a potential use of this enzyme for quantitative spectrophotometric determination of NO and as a component of novel NO sensitive electrodes.

Amino Acid Sequence↗

Ozonated water extends the shelf life of fresh-cut lettuce.

The use of ozonated water as a sanitizer to extend the shelf life of fresh-cut lettuce and the effect on the antioxidant constituents (polyphenols and vitamin C) were investigated. Fresh-cut iceberg lettuce (Lactuca sativa L.) was washed at 4 degrees C using three different ozonated water dips [10, 20, and 10 activated by ultraviolet C (UV-C) light mg L(-1) min total ozone dose], and the dips were compared with water and chlorine rinses. Treated lettuce was packaged in air or active modified atmosphere packaging (MAP) (4 kPa of O2 + 12 kPa of CO2 balanced with N2) and stored for 13 days at 4 degrees C. Despite its strong oxidizing activity, ozonated water did not stimulate the respiratory activity of fresh-cut lettuce. Moreover, ozonated water maintained the initial visual appearance of fresh-cut lettuce and controlled browning during storage in air. Initially, ozonated water and chlorine reduced the total mesophilic population by 1.6 and 2.1 log, respectively, when compared with water. Active MAP was effective in controlling total microbial growth, achieving 2.0 log reduction in relation to samples stored in air at the end of storage. On the other hand, active MAP caused a 2.0-3.5 reduction of coliforms on sanitized samples compared with water-washed samples. The most efficient treatments were ozone 20 and ozone 10 activated by UV-C, which were as effective as chlorine. Changes in individual phenolic compounds were independent of the washing treatments. In air, chlorogenic and isochlorogenic acid contents increased noticeably after 13 days while monocaffeoyltartaric and dicaffeoyltartaric acids remained unchanged. MAP effectively suppressed accumulation of caffeoylquinic derivatives, whereas caffeoyltartaric derivatives decreased during MAP storage to reach similar levels. The content of vitamin C (ascorbic acid and dehydroascorbic acid) decreased during storage, particularly under MAP. Ozonated water could be an alternative sanitizer to chlorine for fresh-cut lettuce due to good retention of sensorial quality and browning control with no detrimental reduction in the antioxidant constituents.

Ascorbic Acid↗

Chitosan treatment of wheat seeds induces resistance to Fusarium graminearum and improves seed quality.

Chitosan treatment (2-8 mg/mL) of wheat seeds significantly improved seed germination to recommended seed certification standards (>85%) and vigor at concentrations >4 mg/mL, in two cultivars of spring wheat (Norseman and Max), by controlling seed-borne Fusarium graminearum infection. The germination was <80% in the control and >85% in benomyl- and chitosan-treated seeds. Seed-borne F. graminearum was reduced to >50% at higher chitosan treatments compared to the control. Synthesis of phenolic acids was stimulated in primary leaves following chitosan treatment, and levels of these phenolic acids, especially ferulic acid, increased significantly with increasing chitosan concentration. Lignin content of primary leaves also showed a similar pattern. The synthesis of precursors of lignin such as p-coumaric, ferulic, and sinapic acids and phenolic acids having antimicrobial activity such as benzoic, p-coumaric, caffeic, protocatechuic, chlorogenic, ferulic, and gallic acids was also stimulated by chitosan treatment. The induction of phenolic acids and lignin was significantly lower in cv. Max compared to Norseman. Chitosan also inhibited fungal transmission to the primary roots of germinating seedlings. Results suggest that chitosan controlled seed-borne F. graminearum infection and increased the resistance in seedlings by stimulating the accumulation of phenolics and lignin. Thus, chitosan has a potential for improvement of seed quality and enhancement of crop yields as well as increased value of stored grains for food and feed.

Benomyl↗

Purification and Characterization of Polyphenol Oxidase from Glandular Trichomes of Solanum berthaultii.

Type A glandular trichomes of the wild potato (Solanum berthaultii Hawkes) entrap insects by rapidly polymerizing the trichome contents after breakage by insect contact. Polymerization of trichome exudate appears to be driven by a soluble polyphenol oxidase (PPO). PPO constitutes up to 70% of the protein in individually collected trichomes and reaches a concentration approaching 200 mum in these organs. Trichome PPO has been purified and shown to be a monomeric copper metalloprotein with an isoelectric point of 5.5, possessing only o-diphenol oxygen oxido-reductase activity, and is larger than most other reported PPOs, with relative molecular weight of 59,000. Chlorogenic and caffeic acid were the most readily oxidized of 14 phenolic substrates tested. Polyclonal antibodies raised against the relative molecular weight 59,000 S. berthaultii trichome PPO were used to show that S. tuberosum L. trichomes express low levels of a cross-reactive protein that lacks detectable PPO activity.

Journal Article↗

Phenols and antioxidative status of Raphanus sativus grown in copper excess.

Raphanus sativus L. cv. Rimbo was grown for 10 days after emergence in hydroponic culture containing 0.12 (control), 5, 10 and 15 micro M copper. The seeds were germinated in the presence of the copper solution. The Cu contents increased with the treatment in both shoots and roots, maintaining in the roots a value eight- to ten-fold higher than in the shoots. With the treatment both shoots and roots underwent growth inhibition and an increase in the percentage of dry weight. Membrane damage and lipid peroxidation increased and glutathione was oxidized as the copper concentration increased, indicating an acceleration of oxidative processes. Control shoots had high contents of reduced glutathione and low contents of phytochelatin-SH whereas roots showed an opposite pattern, suggesting an utilization of reduced glutathione for phytochelatin synthesis. In both parts phytochelatin-SH content reached the maximum at 5 micro M copper and then decreased, reaching at 15 micro M copper the control value in the roots and a value five-fold higher than the control value in the shoots. The main phenolic acids represented in R. sativus were chlorogenic, vanillic, caffeic, siringic, p-coumaric and ferulic acids whereas the least represented were gallic, protocatechuic and p-hydroxybenzoic acids. The phenolic acids as well as the total and reduced ascorbate contents increased with the intensification of copper treatment. Notwithstanding these changes, total ascorbate remained 35% higher in the shoots than in the roots. The reduced ascorbate was thus able to replenish reducing equivalents to phenoxyl radicals thus explaining the increase in the phenolic compounds.

Journal Article↗

Binding of selected phenolic compounds to proteins.

In the context of this study, the noncovalent binding of selected phenolic compounds (chlorogenic, ferulic, and gallic acids, quercetin, rutin, and isoquercetin) to different proteins (human serum albumin, bovine serum albumin, soy glycinin, and lysozyme) was studied with direct (Hummel-Dreyer/size exclusion chromatography) and/or indirect methods (fluorescence absorbance properties of the binding components). In the latter case, the measurement of the phenol binding was achieved by exploiting the intrinsic fluorescence emission properties of quercetin as a probe. From the data obtained, the binding constants and the number of binding sites were calculated. The binding parameters were influenced by different factors, where, e.g., increasing temperature and ionic strength as well as decreasing pH cause a diminished binding. The structures of the proteins as determined by circular dichroism indicate changes in the tertiary structure with the secondary structure remaining intact.

Animals↗

Physicochemical properties and susceptibility to proteolytic digestion of myoglobin-phenol derivatives.

This paper deals with the interactions of chlorogenic, caffeic, and quinic acids and p-quinone with myoglobin. The myoglobin derivatives formed have been characterized in terms of physicochemical properties and susceptibility to proteolysis. The results show that the free amino group and tryptophan contents of the myoglobin-phenol derivatives decrease with the increasing extent to which the protein becomes derivatized. Furthermore, the solubility of myoglobin-phenol derivatives decreases in the pH range 3.5-6.5 as compared to solubility of the native protein. The reaction also influences the hydrophilic-hydrophobic character of the protein. The isoelectric point of the derivatized myoglobin is shifted to a lower pH value, and formation of high molecular fractions is also documented. This paper also demonstrates the influence of the protein derivatization with plant phenols on susceptibility to digestion by trypsin, alpha-chymotrypsin, and pepsin, determined in vitro. The enzymatic digestion of the derivatized proteins is adversely affected.

Hydrolysis↗

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.

Anthocyanins↗