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Protein quality in cereals and pulses. 2. Influence of polyethyleneglycol on the nutritional availability of methionine in sorghum (Sorghum vulgar Pers.), field beans (Vicia faba L.) and barley.

1. Polyethyleneglycol (PEG 4000) was examined for its influence on relative nutritional value (RNV) and available methionine in sorghum (Sorghum vulgare Pers.), field beans (Vicia faba L.) and barley, as measured microbiologically with Streptococcus zymogenes. The results were assessed in relation to the content of tannins in the test samples. 2. In grain of hybrid sorghum the RNV averaged 87 (range 79--92) for six low-tannin varieties and 41 (30--53) for eleven high-tannin varieties. The corresponding available methionine values averaged 17.0 (15.7--18.9) and 8.9 (6.7--11.0) g/kg protein. Addition of PEG 4000 to the test samples increased the average RNV of the high-tannin varieties from 41 to 78, and the average available methionine content from 8.9 to 16.2 g/kg protein. 3. With seed of ten coloured flowered varieties of field beans, treatment with PEG gave a small but consistent increase in the available methionine content, which resulted from the inactivation of tannins in the testa. 4. In twenty-three samples of barley grain, treatment with PEG had no effect on the values obtained for available methionine. 5. Treatment of high-tannin sorghum grain with ammonia has been reported to inactivate the tannins and increase the nutritional value for rats and chicks. This finding was confirmed. The present study showed that ammonia and PEG 4000 were equally effective in enhancing the nutritional quality as measured in the microbiological tests.

Dietary Proteins↗

Chemical, biochemical, and biological significance of polyphenols in cereals and legumes.

Polyphenols in cereals and legumes have been receiving considerable attention largely because of their adverse influence on color, flavor, and nutritional quality. These compounds belong to the flavonoid and tannin groups and are mostly located in the seed coat or pericarp of the grains. The pearl millet flavonoids have been identified as C-glycosylflavones by the combined use of paper chromatography and UV spectroscopy. Although nontoxic, physiological and nutritional significance of these compounds occurring in high amounts in the pearl millet grain are still not clearly understood. In view of aesthetic quality, bleaching of the millet grains in acidic solution is recommended. A large proportion of current assays involves spectrophotometry of tannin or its chromogen and tannin-protein interaction. Sorghum and legume tannins have been characterized as condensed tannins. Several factors such as plant type, age of the plant or plant parts, stage of development, and environmental conditions govern the polyphenol contents in plants. Polyphenols are known to interact with proteins and form tannin-protein complexes leading to either inactivation of enzymes or making proteins insoluble. These are implicated in decreasing the activities of digestive enzymes, protein and amino acid availabilities, mineral uptake, vitamin metabolism, and depression of growth. Polyphenols are known to cause certain ultrastructural changes in the different parts of experimental animals. A correlation between dietary tannins and occurrence of esophageal cancer has been established. Bird resistance and seed germination in food crops have been correlated to high contents of polyphenols. The antinutritional activity of polyphenols can be reduced by removing polyphenols from the grains by chemical treatments or removing pericarp and testa by pearling. Treatment of alkaline reagents and ammonia can remove 90% of the polyphenols. Supplementation of polyphenols-rich diet with protein can alleviate the growth-depressing effect of polyphenols.

Chemical Phenomena↗

Migration of Calcium and Its Role in the Regulation of Seismonasty in the Motor Cell of Mimosa pudica L.

Volume and conformational changes of the contractile tannin vacuoles of the abaxial motor cells of the primary pulvinus of Mimosa pudica L. parallel the seismonastic leaf movement. Since such changes in cells and organelles of animal systems are often regulated by calcium, we studied Ca(2+) movement in the motor cells and tissue. By fixation with Lillie's neutral buffered formalin, followed by staining with alizarin red sulfate (ARS), calcium was localized in the tannin vacuoles of the motor cells of the primary pulvinus. After treatment with ethylenediaminetetraacetate, 8-hydroxyquinoline, and several other calcium-complexing or extracting agents, the color reaction due to alizarin red sulfonate was no longer present. By using an analytical method, it was shown that the effluent from stimulated pulvini has significantly more Ca(2+) than that from unstimulated controls. Ten millimolar LaCl(3) inhibits recovery of the tannin vacuole in vivo in 10 mm CaCl(2) or in distilled water. Quantitative data obtained by microspectrophotometry demonstrated calcium migration during the bending movement of the primary pulvinus. In the adaxial motor cells a small amount of calcium migrates from the tannin vacuole, and calcium on the cell wall moves to the central vacuole. In the abaxial half, a large amount of calcium from the tannin vacuole moves to the central vacuole of the motor cell. It is probable that the calcium binds to the microfibrillar contents of the central vacuole. These observations support the contention that Ca(2+) migrates between the surface of the tannin vacuole and the inside of the central vacuole. The recovery and maintenance of the tannin vacuole in the spherical form may play a role in maintaining turgor in the motor cells of the abaxial half of the primary pulvinus of Mimosa.

Journal Article↗

EFFECT OF CAROB POD EXTRACT ON CELLULOLYSIS, PROTEOLYSIS, DEAMINATION, AND PROTEIN BIOSYNTHESIS IN AN ARTIFICIAL RUMEN.

Carob pod extract and its tannin and sugar fractions were compared with gallotannic acid and sucrose for their effect on the cellulolytic, proteolytic, protein biosynthetic, and deaminative activities of rumen microorganisms. The inhibitory effects of carob pod extract upon the cellulolysis and deamination were correlated mainly with its sugar, rather than its tannin components. On the other hand, proteolytic activity and protein biosynthesis were more significantly affected by the tannin fraction. In contrast to the tannin fraction of carob pod extract, gallotannic acid inhibited cellulolytic activity. The harmful effect of a low concentration of tannins on protein biosynthesis could be prevented by the addition of carbohydrates to the reaction mixture. At high tannin concentration (40 mug/ml), however, the addition of carbohydrates did not prevent the inhibition.

Ammonia↗

Relationship between phospholamban and nucleotide activation of cardiac sarcoplasmic reticulum Ca2+ adenosinetriphosphatase.

A strong connection with nucleotide activation of Ca2+ATPase and phospholamban inhibition has been found. Phospholamban decreases the number of activatable Ca2+ATPase without affecting substrate affinity or the ability of nucleotide to serve its dual modulatory roles, i.e., catalytic and regulatory. Low concentrations of certain nucleotide mimetics, quercetin, tannin, and ellagic acid, with structural similarity to adenine can unmask phospholamban's inhibitory effect while concurrently acting as competitive inhibitors of nucleotide binding. Micromolar concentrations of tannin (EC50 approximately 0.3 microM) and ellagic acid (EC50 approximately 3 microM) stimulated Ca2+ uptake and calcium-activated ATP hydrolysis at submicromolar Ca2+ in isolated cardiac sarcoplasmic reticulum (SR). Stimulation of Ca2+ATPase was followed by pronounced inhibiton at only slightly higher tannin concentrations (IC50 approximately 3 microM), whereas inhibitory effects by ellagic acid were observed at much greater concentrations (IC50 > 300 microM) than the EC50. A complex relationship between compound, SR protein, and MgATP concentration is a major determining factor in the observed effects. Stimulation was only observed under conditions of phospholamban regulation, while the inhibitory effects were observed in cardiac SR at micromolar Ca2+ and in skeletal muscle SR, which lacks phospholamban. Maximal stimulation of Ca2+ATPase was identical to that observed with the anti-phospholamban monoclonal antibody 1D11. Both compounds appear to relieve the Ca2+ATPase from phospholamban inhibition, thereby increasing the calcium sensitivity of the Ca2+ATPase like that observed with phosphorylation of phospholamban or treatment with monoclonal antibody 1D11. Tannin, even under stimulatory conditions, is a competitive inhibitor of MgATP with a linear Dixon plot. The subsequent inhibitory action of higher tannin concentrations results from competition of tannin with the nucleotide binding site of the Ca2+ATPase. In contrast, ellagic acid produced a curvilinear Dixon plot suggesting partial inhibition of nucleotide activation. The data suggest that nucleotide activation of Ca2+ATPase is functionally coupled to the phospholamban interaction site. These compounds through their interaction with the adenine binding domain of the nucleotide binding site prevent or dissociate phospholamban regulation. Clearly, this portion of Ca2+ATPase needs further study to elucidate its role in phospholamban inhibition.

Adenosine Triphosphate↗

Effects of different organic farming methods on the concentration of phenolic compounds in sea buckthorn leaves.

The effects of different cultivation methods on the amount of phenolic compounds in leaves of 1-year-old seedlings of two Finnish sea buckthorn (Hippophae rhamnoides L. ssp. rhamnoides) cultivars 'Terhi' and 'Tytti' were studied in a field experiment established at coastal area in Merikarvia, western Finland. The cultivation methods included different fertilizers (suitable for organic cultivation), mulches (organic and plastic), and land contours (flat vs low hill surface). Two experiments were conducted. The first allowed the estimation of the effects of cultivar, fertilizer, surface contour, and all their interactions, while the other allowed the estimation of the effects of mulches, land contours, and their interactions for the cultivar 'Tytti'. Eleven different hydrolyzable tannins, pentagalloylglucose, and 14 other phenolic compounds were detected by chemical analysis with high-performance liquid chromatography (HPLC). The amount of phenolic compounds varied between different land contours and mulches. The concentrations of gallic acid, pentagalloylglucose, quercetin-3-rhamnoside, monocoumaroyl astragalin A, total hydrolyzable tannins, and condensed tannins were significantly higher on the flat surface than on the low hill surface. The plastic mulch decreased the concentration of gallic acid, hydrolyzable tannins, and condensed tannins compared to the other mulches used. These results suggest ways to cultivate sea buckthorn to produce large amounts of valuable chemicals, especially tannins in the leaves.

Agriculture↗

Removal of dissolved brown algal phlorotannins using insoluble polyvinylpolypyrrolidone (PVPP).

Tannins, a large and diverse group of phenolic secondary metabolites, are common in terrestrial plants and marine brown algae. It is sometimes desirable to remove the tannins from plant or algal extracts, e.g., when isolating enzymes and nucleic acids, when using certain colorimetric methods to quantify the tannin content, or to create reliable controls when using tannins in experimental studies. Insoluble polyvinylpolypyrrolidone (PVPP) can be used to specifically remove tannins from solution. In the present study, we evaluated the effect of different factors (amount of PVPP, number of PVPP treatments, type of solvent, pH, and incubation time) on the PVPP removal of dissolved brown algal phlorotannins. Our results imply that there is a limited amount of phlorotannins that can bind to a given amount of PVPP, and that it is preferable to use low quantities of PVPP repeatedly, compared to using fewer treatments with a high amount of PVPP. Furthermore, we found no consistent effect on the removal of phlorotannins due to solvent type (acetone, methanol, distilled water or filtered seawater). There was a slight decrease in the amount of phlorotannins removed from extracts with increasing pH when repeatedly treated with PVPP. All phlorotannins were removed from extracts with pH < or = 6.2, and 89% of the initial phlorotannin content was removed at pH 9.7. These results are compared with previous methodological studies on tannin removal with PVPP. Furthermore, the implications of phlorotannin removal in analytical and ecological investigations are discussed.

Biological Assay↗

Differential activities of plant polyphenols on the binding and internalization of cholera toxin in vero cells.

Plant polyphenols, RG-tannin, and applephenon had been reported to inhibit cholera toxin (CT) ADP-ribosyltransferase activity and CT-induced fluid accumulation in mouse ileal loops. A high molecular weight fraction of hop bract extract (HBT) also inhibited CT ADP-ribosyltransferase activity. We report here the effect of those polyphenols on the binding and entry of CT into Vero cells. Binding of CT to Vero cells or to ganglioside GM1, a CT receptor, was inhibited in a concentration-dependent manner by HBT and applephenon but not RG-tannin. These observations were confirmed by fluorescence microscopy using Cy3-labeled CT. Following toxin binding to cells, applephenon, HBT, and RG-tannin suppressed its internalization. HBT or applephenon precipitated CT, CTA, and CTB from solution, creating aggregates larger than 250 kDa. In contrast, RG-tannin precipitated CT poorly; it formed complexes with CT, CTA, or CTB, which were demonstrated with sucrose density gradient centrifugation and molecular weight exclusion filters. In agreement, CTA blocked the inhibition of CT internalization by RG-tannin. These data suggest that some plant polyphenols, similar to applephenon and HBT, bind CT, forming large aggregates in solution or, perhaps, on the cell surface and thereby suppress CT binding and internalization. In contrast, RG-tannin binding to CT did not interfere with its binding to Vero cells or GM1, but it did inhibit internalization.

Animals↗

Effect of cultivars on in vitro and ruminal degradation of the nitrogen fraction in birdsfoot trefoil silage.

Experiments were carried out to determine the effects of tannin content on N fractions and DM digestibility of birdsfoot trefoil harvested at 20% bloom stage. Five trefoil cultivars, harvested in both June and August, were used: Upstar, Mirabel, Empire, Leo, and Bt-8861. Forage was ensiled in 1.3-kg laboratory silos for 60 d with 4 silos per cultivar and harvest. Concentrations of DM and tannin in forage were determined prior to ensiling, and N fractions, DM digestibility, and pH in silage were also determined. Tannin concentration and protein N content of silage were lower in Empire than in the other cultivars. Ruminal degradability of DM and CP of Empire (low tannin) and Upstar (high tannin) birdsfoot trefoil were estimated with two fistulated cows using nylon bags that were incubated < or = 96 h. The DM-soluble fraction of Empire was significantly lower than that of Upstar for the first harvest only. This difference was partly due to the high proportions of dry shoots. The CP-soluble fraction of Empire was higher than that of Upstar, but the percentage of CP that was potentially degradable in the rumen was lower for Empire. This difference could result in different CP availability over time even for similar effective degradability of CP. Tannin had no additional effect on total CP degradability in the rumen.

Animals↗

EFFECT OF WATER EXTRACTS OF CAROB PODS, TANNIC ACID, AND THEIR DERIVATIVES ON THE MORPHOLOGY AND GROWTH OF MICROORGANISMS.

The effect of aqueous extracts of carob (Ceratonia siliqua) pods, gallotannic acid, gallic acid, and catechol on several microorganisms was studied. Carob pod extract and tannic acid showed a strong antimicrobial activity toward some cellulolytic bacteria. On the basis of tannin content, to which antimicrobial effect was related, carob pod extracts inhibited Cellvibrio fulvus and Clostridium cellulosolvens at 15 mug/ml, Sporocytophaga myxococcoides at 45 mug/ml, and Bacillus subtilis at 75 mug/ml. The inhibiting concentrations for tannic acid were found to be 12, 10, 45, and 30 mug/ml, respectively. Gallic acid and catechol were much less effective. Tannic acid and the tannin fraction of carob extract exerted both bacteriostatic and bactericidal effects on C. fulvus. Respiration of C. fulvus in the presence of bactericidal concentrations of tannic acid or tannin fraction of carob extract was inhibited less than 30%. A partial formation of "protoplasts" by C. fulvus was obtained after 2 hr of incubation in a growth medium to which 20% sucrose, 0.15% MgSO(4).7H(2)O, and 10 to 50 mug/ml of tannic acid or 500mug/ml of penicillin, or both, had been added. Tannic acid and the tannin fraction of carob extract protected C. fulvus from metabolic lysis in sucrose solution. Although the growth of other microorganisms tested was only slightly affected, the morphology of some of them was drastically changed in the presence of subinhibitory concentrations of carob pod extracts of tannic acid. It is suggested that the site of action of tannins on sensitive microorganisms is primarily the cell envelope.

Bacillus subtilis↗

Resistance and tolerance of Terminalia sericea trees to simulated herbivore damage under different soil nutrient and moisture conditions.

Resource availability, degree of herbivore damage, genetic variability, and their interactions influence the allocation of investment by plants to resistance and tolerance traits. We evaluated the independent and interactive effects of soil nutrients and moisture, and simulated the effects of herbivore damage on condensed tannins (resistance) and growth/regrowth (tolerance) traits of Terminalia sericea, a deciduous tree in the Kalahari desert that constitutes a major component of livestock diet. We used a completely crossed randomized-block design experiment to examine the effects of nutrients, water availability, and herbivore damage on regrowth and resistance traits of T. sericea seedlings. Plant height, number of branches, internode length, leaf area, leaf mass for each seedling, combined weight of stems and twigs, and root mass were recorded. Condensed tannin concentrations were 22.5 and 21.5% higher under low nutrients and low soil moisture than under high nutrient and high water treatment levels. Tannin concentrations did not differ significantly between control and experimental seedlings 2 mo after simulated herbivore damage. Tannin concentrations correlated more strongly with growth traits under low- than under high-nutrient conditions. No trade-offs were detected among individual growth traits, nor between growth traits and condensed tannins. T. sericea appeared to invest more in both resistance and regrowth traits when grown under low-nutrient conditions. Investment in the resistance trait (condensed tannin) under high-nutrient conditions was minimal and, to a lesser degree, correlated with plant growth. These results suggest that T. sericea displays both resistance and tolerance strategies, and that the degree to which each is expressed is resource-dependent.

Adaptation, Biological↗

The molecular interaction of human salivary histatins with polyphenolic compounds.

Dietary tannins are polyphenols that are effectively precipitated by salivary histatins (Hsts), a novel family of tannin binding proteins. Epigallocatechin gallate (EGCG), a flavan-3-ol ester related to condensed tannins (polymerized products of flavan-3-ols), and pentagalloyl glucose (PGG), a hydrolyzable tannin, were used to evaluate the molecular nature of Hst-polyphenol interaction. NMR demonstrated that Hst5, a representative Hst, bound to EGCG in a hydrophobic manner via basic and aromatic residues. In contrast, proline plays a dominant role in polyphenol binding to other tannin precipitating proteins. The role of basic and aromatic amino acids in EGCG binding was investigated using a series of modified Hsts in each of which one type of amino acid was substituted by Ala. EGCG bound to all modified Hsts, but the binding was diminished. Optimal EGCG binding also depended on the primary structure, as a polypeptide with randomised Hst5 sequence showed significantly diminished interaction with EGCG. Soluble EGCG/Hst5 complexes containing up to seven molecules of EGCG per mol of Hst5 had a 1-mM dissociation constant. In contrast to EGCG, PGG formed small soluble complexes with Hst5 consisting of only one molecule each of PGG and Hst5, as demonstrated by analytical ultracentrifugation. These complexes became insoluble upon binding of additional molecules of PGG. Diminished PGG binding was seen to a peptide with a Hst5 randomized sequence showing the importance of the primary structure. Hsts may serve to form insoluble complexes with tannins thereby preventing their absorption from the intestines and potentially harmful biological effects. In contrast the much weaker interaction with EGCG may allow its uptake into the organism and exploitation of its antioxidant effect.

Amino Acid Sequence↗

Overview of dietary flavonoids: nomenclature, occurrence and intake.

Flavonoids and their polymers constitute a large class of food constituents, many of which alter metabolic processes and have a positive impact on health. Flavonoids are a subclass of polyphenols. They generally consist of two aromatic rings, each containing at least one hydroxyl, which are connected through a three-carbon "bridge" and become part of a six-member heterocyclic ring. The flavonoids are further divided into subclasses based on the connection of an aromatic ring to the heterocyclic ring, as well as the oxidation state and functional groups of the heterocyclic ring. Within each subclass, individual compounds are characterized by specific hydroxylation and conjugation patterns. Many flavonoids in foods also occur as large molecules (tannins). These include condensed tannins (proanthocyanidins), derived tannins and hydrolysable tannins. For proanthocyanidins, three subclasses (15 characterized) have been identified in foods. Monomers are connected through specific carbon-carbon and ether linkages to form polymers. Derived tannins are formed during food handling and processing, and found primarily in black and oolong teas. Flavonoids are widely distributed in nature, albeit not uniformly. As a result, specific groups of foods are often rich sources of one or more subclasses of these polyphenols. The polyphenolic structure of flavonoids and tannins renders them quite sensitive to oxidative enzymes and cooking conditions. Scientists in several countries have estimated intakes of a few subclasses of flavonoids from limited food composition databases. These observations suggest large differences in consumption, due in part to cultural and food preferences among populations of each country.

Diet↗

The specificity of proanthocyanidin-protein interactions.

The proanthocyanidins or condensed tannins, phenolic polymers which are synthesized by many plants, characteristically bind and precipitate proteins. The specificity of the interaction was investigated using a competitive binding assay to compare directly the affinities of various proteins and synthetic polymers for the tannin obtained from Sorghum bicolor (Lin.) Moench. At pH 4.9, the relative affinities range over more than 4 orders of magnitude, indicating that this proanthocyanidin interacts quite selectively with protein and protein-like polymers. The affinity for tannins is an inverse function of the size of the polymer, and peptides with less than six residues interact very weakly with tannin. Proteins are precipitated by proanthocyanidins most efficiently at pH values near their isoelectric points. Proline-rich proteins and polymers have very high affinities for tannin. Tightly coiled globular proteins have much lower affinities for tannin than conformationally loose proteins.

Amino Acids↗

Tannic acid inhibits insulin-stimulated lipogenesis in rat adipose tissue and insulin receptor function in vitro.

Tannins occur naturally in relatively abundant amounts in fruits, herbal medicines and common beverages. Thus an understanding of how these polyphenols affect peptide hormone action is of importance. We report here that tannic acid (a hydrolysable tannin) inhibits insulin-stimulated lipogenesis in rat adipose tissue in vitro, with an IC50 estimated to be about 350 microM. However, its monomer, gallic acid, did not show a similar inhibitory effect at concentrations up to 1 mM. The inhibition by tannic acid was less evident with higher concentrations of bovine serum albumin in the incubation buffer. This was attributed to the formation of a tannin-protein complex between bovine serum albumin and tannic acid. In a binding assay, it was observed that the specific binding of insulin to its receptor was not inhibited by tannic acid in the concentration range 0-200 microM. However, insulin-stimulated autophosphorylation of the insulin receptor, and receptor-associated tyrosine kinase phosphorylation of RR-SRC peptide, were inhibited by tannic acid at concentrations as low as 25 microM. Our data do not support the current speculation that tannins affect the activity of peptide hormones by binding to them. Therefore, our finding opens up a new perspective in the understanding of the mode of action of tannins on such hormones.

Adipocytes↗

Decreased polyphenol transport across cultured intestinal cells by a salivary proline-rich protein.

Tannins are polyphenols commonly found in plant-derived foods. When ingested they can have various harmful effects, but salivary proline-rich proteins (PRPs) may provide protection against dietary tannins. The aim of this study was to investigate whether basic PRPs, a major family of salivary proteins, can prevent intestinal absorption of tannin. To do so it was necessary first to characterize transport of pentagalloyl glucose (5GG), a hydrolysable tannin, across cultured epithelial cells. Using human intestinal epithelial cells (Caco-2 cells) it was found that a partial degradation of 5GG occurred during transepithelial transport resulting in the presence of 5GG as well as tetra- and trigalloyl glucose and glucose in the receiving compartment. The sodium-dependent glucose transporter SGLT1 played a role in apical (mucosal) to basolateral (serosal) transport and transport in the opposite direction was dependent on the multidrug resistance-associated protein MRP2. An increased uptake from the apical compartment was seen when the basolateral receiving solution was human serum rather than a balanced salt solution. Transport both in apical-basolateral and basolateral-apical directions was reduced when 1B4, a human basic PRP, was added to the 5GG-containing medium. This decrease closely paralleled the formation of insoluble 5GG-1B4 complexes. It appears that the formation of insoluble tannin-protein complexes diminishes the uptake of 5GG and its metabolites. There is little evidence of other biological activities of basic PRPs so in contrast to other salivary proteins they may exert a biological function in the intestines.

Apolipoprotein A-I↗

Analysis of commercial vegetable tanning agents by reversed-phase liquid chromatography-electrospray ionization-tandem mass spectrometry and its application to wastewater.

Commercial vegetable tanning agents that are derived from plants and consist of condensed or hydrolyzable tannins were analyzed by electrospray ionization-tandem mass spectrometry (ESI-MS/MS) to identify their major constituents and to study their collision-induced dissociation. In the condensed tannin wattle a series of proanthocyanidin dimers to tetramers was identified together with the flavonoid monomers catechin and gallocatechin. The composition of the hydrolyzable tannin chestnut was more heterogenous. Besides the monomers ellagic and gallic acid a variety of gallotannins were detected, namely mono-, di- and trigalloylglucose, and a variety of ellagitannins. Reversed-phase HPLC-ESI-MS/MS methods were developed to detect condensed and hydrolyzable tannins in tannery wastewaters by multiple reaction monitoring (MRM). The methods proved suitable even for highly loaded wastewaters. However, the detected amount of wattle tanning agent in spent retanning baths was about two orders of magnitude below the amount used for the retanning. This suggests that the condensed tannins of polyphenolic structure are rapidly transformed during the tanning process to yet unknown products.

Chromatography, High Pressure Liquid↗

Direct scavenging of nitric oxide and superoxide by green tea.

In the present study, we investigated the free radical scavenging effects of green tea extract and green tea tannin mixture and its components using a nitric oxide (NO) and superoxide (O(2)(-)) generating system in vitro. Green tea extract showed direct scavenging activity against NO and O(2)(-) and green tea tannin mixture, at the same concentration, showed high scavenging activity. Comparison of the activities of seven pure compounds isolated from green tea tannin mixture showed that (-)-epigallocatechin 3-O-gallate (EGCg), (-)-gallocatechin 3-O-gallate (GCg) and (-)-epicatechin 3-O-gallate (ECg) had higher scavenging activities than (-)-epigallocatechin (EGC), (+)-gallocatechin (GC), (-)-epicatechin (EC) and (+)-catechin (C), showing the importance of the structure of flavan-3-ol linked to gallic acid for this activity. Among the gallate-free tannins, EGC and GC were more effective O(2)(-) scavengers than EC and C, indicating the O-trihydroxy structure in the B ring is an important determinant of such activity. However, this structure did not affect the NO scavenging activity. These findings confirm that green tea tannin has excellent antioxidant properties, which may be involved in the beneficial effect of this compound.

Catechin↗