Radical scavenging capacity of wine anthocyanins is strongly pH-dependent

The radical scavenging capacity of red wine anthocyanins was quantified by the so-called TEAC assay with special emphasis on the influence of pH and conjugation on this activity. The pH appears to be a dominant factor in the radical scavenging capacity of wine anthocyanins, with higher pH values increasing this capacity significantly. On the basis of the pKa values for deprotonation and theoretical calculations, it could be concluded that the effect is due to an increase in intrinsic radical scavenging capacity upon deprotonation. The data also reveal that the reduction in radical scavenging activity of anthocyanins upon their conjugation can, at least in part, be ascribed to an increase in pKa values upon conjugation. Altogether, the results obtained provide molecular insight into factors that influence radical scavenging potential of anthocyanins and reveal that the radical scavenging-mediated supposed beneficial health effects of these wine pigments will be influenced by the pH of the surrounding matrix or tissue.     

Factors affecting the levels of tea polyphenols and caffeine in tea leaves

An isocratic HPLC procedure was developed for the simultaneous determination of caffeine and six catechins in tea samples. When 31 commercial teas extracted by boiling water or 75% ethanol were analyzed by HPLC, the levels of (-)-epigallocatechin 3-gallate (EGCG), and total catechins in teas were in the order green tea (old leaves) > green tea (young leaves) and oolong tea > black tea and pu-erh tea. Tea samples extracted by 75% ethanol could yield higher levels of EGCG and total catechins. The contents of caffeine and catechins also have been measured in fresh tea leaves from the Tea Experiment Station in Wen-Shan or Taitung; the old tea leaves contain less caffeine but more EGCG and total catechins than young ones. To compare caffeine and catechins in the same tea but manufactured by different fermentation processes, the level of caffeine in different manufactured teas was in the order black tea > oolong tea > green tea > fresh tea leaf, but the levels of EGCG and total catechins were in the order green tea > oolong tea > fresh tea leaf > black tea. In addition, six commercial tea extracts were used to test the biological functions including hydroxyl radical scavenging, nitric oxide suppressing, and apoptotic effects. The pu-erh tea extracts protected the plasmid DNA from damage by the Fenton reaction as well as the control at a concentration of 100 microg/mL. The nitric oxide suppressing effect of tea extracts was in the order pu-erh tea >/= black tea > green tea > oolong tea. The induction of apoptosis by tea extract has been demonstrated by DNA fragmentation ladder and flow cytometry. It appeared that the ability of tea extracts to induce HL-60 cells apoptosis was in the order green tea > oolong > black tea > pu-erh tea. All tea extracts extracted by 75% ethanol have stronger biological functions than those extracted by boiling water.     

Protection against nitric oxide toxicity by tea

It is found that green tea and black tea are able to protect against nitric oxide (NO(*)) toxicity in several ways. Both green tea and black tea scavenge NO(*) and peroxynitrite, inhibit the excessive production of NO(*) by the inducible form of nitric oxide synthase (iNOS), and suppress the LPS-mediated induction of iNOS. The NO(*) scavenging activity of tea was less than that of red wine. The high activity found in the polyphenol fraction of black tea (BTP) could not be explained by the mixed theaflavin fraction (MTF) or catechins [epicatechin, epigallocatechin, epicatechin gallate, epigallocatechin gallate (EGCG)], which were tested separately. Synergistic effects between the compounds, or the presence of a potent, unidentified NO(*) scavenger, may explain the high activity of BTP. The peroxynitrite scavenging of tea was comparable to that of red wine. The main activity was found in the polyphenol fraction. MTF and the catechins were found to be potent peroxynitrite scavengers. Tea and tea components were effective inhibitors of iNOS. Of the tea components tested, only MTF had an activity higher than that of the tea powders. The polyphenol fractions of tea were much more active than the tea powders in suppressing the induction of iNOS. On the basis of its abundance and activity, EGCG was the most active inhibitor. The protective effect of tea on NO(*) toxicity is discussed in relation to the beneficial effect of flavonoid intake on the occurrence of cardiovascular heart disease.     

Determination of peroxyl radical scavenging activity of flavonoids and plant extracts using an automatic potentiometric titrator

A novel potentiometric method for evaluation of peroxyl radical scavenging activity of flavonoids and plant extracts was developed. The oxidation of potassium iodide (KI) was performed in acetonitrilephosphate buffer (1:1) containing antioxidant using 2,2'-azobis(2-amidinopropane) dihydrochloride as a peroxyl radical generator. The amount of iodine released from KI during a 20-min free radical oxidation was determined quantitatively using an automatic potentiometric titrator with sodium thiosulfate. The radical scavenging activity of the sample was expressed as the inhibition ratio for iodine release of the control group mediated by the radical. The results obtained from some authentic polyphenols correlated well with those of previous reports. This is a simple, time-saving method requiring less than 30 min and is useful in assessing the radical scavenging activity of antioxidants in plant extracts. We describe the radical scavenging activities of various flavonoids including 21 kinds of tea catechins and vegetable extracts by this method.     

Effects of pu-erh tea on oxidative damage and nitric oxide scavenging

The effects of pu-erh tea, which is prepared by fermentation of tea, on oxidative damage and nitric oxide scavenging, compared with various other brands of tea were investigated. The total antioxidant activity was determined using the Trolox equivalent antioxidant capacity (TEAC) assay. The results showed that TEAC values of the 200 microg/mL water extracts of pu-erh tea (WEPT), green tea, oolong tea, and black tea were 86.3, 85.3, 87.4, and 80.3 (microg/mL), respectively, indicating that WEPT showed a significant antioxidant activity. WEPT, like green tea extract, oolong tea extract, and black tea extract, exhibited a remarkable protective effect in lipid (liposome) and nonlipid (protein and deoxyribose) model systems, implying that it is an inhibitor of lipid and nonlipid oxidative damage. It also exhibited metal-binding ability, reducing power, and scavenging effect for free radicals. Moreover, WEPT showed a decreasing effect on nitric oxide production of lipopolysaccharide-induced RAW 264.7 macrophages. In addition, the results revealed that epicatechin (EC), flavonoid, ascorbic acid, and polyphenolic compounds are present in WEPT, which may partially account for the protective effect on oxidative damage. Thus, WEPT may have potential as an antioxidant and as a nitric oxide scavenging agent.     

Effect of selenium on increasing the antioxidant activity of tea leaves harvested during the early spring tea producing season

This research was to determine the effect of foliar application of selenium on increasing the antioxidant activity of tea harvested during the early spring tea producing season using a alpha,alpha-diphenyl-beta-picrylhydrazyl (DPPH) radical scavenging method and the linoleic acid system. The results showed that the radical scavenging ability of the tea extracts followed this order during the first 60 min: selenium-enriched tea obtained by fertilization with selenate > BHT > selenium-enriched tea obtained by fertilization with selenite > alpha-tocopherol > regular tea. Se-enriched tea obtained by fertilization with selenate exhibited the highest inhibition percentage of 84.29% at 30 min. Se-enriched tea extracts provided higher hydrogen-donating capabilities than regular tea and contrasts with BHT and alpha-tocopherol at the concentration of 100 microg of solids/mL of ethanol. There was a little change in the sequence of radical scavenging ability during the later 60 min: Se-enriched tea obtained by fertilization with selenate > Se-enriched tea obtained by fertilization with selenite > BHT > regular tea > alpha-tocopherol. The individual activity of tea extracts and references measured by the linoleic acid system showed that the tea extracts, BHT, and alpha-tocopherol manifested almost the same patterns of activity as the DPPH method. Tea enriched in selenium by fertilization with selenate still exhibited the highest inhibition activity of lipid oxidation, whereas alpha-tocopherol showed the lowest inhibition. The antioxidant activity of Se-enriched green tea harvested during the early spring tea producing season is enhanced compared to regular tea.     

Size effect of se-enriched green tea particles on in vitro antioxidant and antitumor activities

The antioxidant and antitumor activities (in vitro) of superfine regular and Se-enriched green tea particles with different sizes (3.52 microm and 220 nm) were investigated in this paper. The vitamin C and tea polyphenol contents of green tea in different sizes were significantly different, and amino acid and chlorophyll just changed a little. The antioxidant activity of green tea particles was evaluated by DPPH radical scavenging and linoleic acid peroxidation inhibition methods, and the antitumor activity was evaluated by antiproliferation assay on HepG2, A549, and MGC803 cells. The results indicated that enrichment of selenium endowed green tea with higher antioxidant activity and antitumor activity on HepG2 and A549 cells but not on MGC803 cells. The DPPH radical scavenging rates of regular and Se-enriched green tea of 220 nm (67.87% and 69.49%, respectively) were significantly greater than that of 3.52 microm, but the inhibition of linoleic acid peroxidation for green tea of 220 nm was lower. The inhibitory rates of green tea of 220 nm on HepG2, A549, and MGC803 cells achieved 77.35%, 80.76%, and 87.54% for regular green tea, and 82.51%, 88.09%, and 74.48% for Se-enriched green tea at the dose of 100 microg mL (-1), values that were all significantly higher compared to that of 3.52 microm.     

Simple assessment of radical scavenging capacity of beverages

The radical-scavenging antioxidants play an important role against oxidative stress in the defense system in vivo. The beneficial effects of antioxidants contained in foods and beverages have been well-accepted, and their antioxidant capacity has been assessed by various methods. In the present study, a simple method is proposed in which the total radical scavenging capacity is assessed from the bleaching of pyranine and pyrogallol red induced by free radicals generated from azo initiator. The total content of antioxidants contained in red wine, green tea, and cassis drink and their reactivities toward peroxyl radicals were measured from the lag phase and rate of bleaching using pyranine and pyrogallol red as a probe, respectively. It was found that this method to follow the bleaching of two probes by visible light spectrophotometer is convenient and applicable for assessment of total radical scavenging capacity of both content and activity of the antioxidants contained in beverages.     

Catechin and caffeine content of green tea dietary supplements and correlation with antioxidant capacity

The health benefits associated with tea consumption have resulted in the wide inclusion of green tea extracts in botanical dietary supplements, which are widely consumed as adjuvants for complementary and alternative medicines. Tea contains polyphenols such as catechins or flavan-3-ols including epicatechin, epigallocatechin, epicatechin gallate, and epigallocatechin gallate (EGCG), as well as the alkaloid, caffeine. Polyphenols are antioxidants, and EGCG, due to its high levels, is widely accepted as the major antioxidant in green tea. Therefore, commercial green tea dietary supplements (GTDS) may be chemically standardized to EGCG levels and/or biologically standardized to antioxidant capacity. However, label claims on GTDS may not correlate with actual phytochemical content or antioxidant capacity nor provide information about the presence and levels of caffeine. In the current study, 19 commonly available GTDS were evaluated for catechin and caffeine content (using high-performance liquid chromatography) and for antioxidative activity [using trolox equivalent antioxidant capacity (TEAC) and oxygen radical antioxidant capacity (ORAC) assays]. Product labels varied in the information provided and were inconsistent with actual phytochemical contents. Only seven of the GTDS studied made label claims of caffeine content, 11 made claims of EGCG content, and five specified total polyphenol content. Caffeine, EGCG, and total polyphenol contents in the GTDS varied from 28 to 183, 12-143, and 14-36% tablet or capsule weight, respectively. TEAC and ORAC values for GTDS ranged from 187 to 15340 and from 166 to 13690 mumol Trolox/g for tablet or capsule, respectively. The antioxidant activities for GTDS determined by TEAC and ORAC were well-correlated with each other and with the total polyphenol content. Reliable labeling information and standardized manufacturing practices, based on both chemical standardization and biological assays, are recommended for the quality control of botanical dietary supplements.     

Role of polyphenol oxidase and peroxidase in the generation of black tea theaflavins.

It has been reported earlier that when macerated tea leaf is fermented at lower pH, the resultant black tea contains increased levels of theaflavin, an important quality marker in black tea. In an attempt to investigate the biochemistry and chemistry underlying this observation, in vitro oxidation experiments using polyphenol oxidase (PPO) from fresh tea leaves, horseradish peroxidase (POD), and tea catechins, precursors for theaflavins, were carried out. In vitro oxidation experiments using crude tea PPO resulted in higher content of theaflavins at pH 4.5 in comparison with pH 5.5, the normal pH of the macerated tea leaf. When purified PPO was used in the in vitro system, surprisingly a reversal of this trend was observed, with more theaflavins being formed at the higher pH. A combination of pure tea PPO and POD led to an observation similar to that with the crude enzyme preparation, suggesting a possible role for POD in the formation or degradation of theaflavin. POD was observed to oxidize theaflavins in the presence of H(2)O(2), leading to the formation of thearubigin, another black tea pigment. This paper demonstrates that tea PPO, while oxidizing catechins, generates H(2)O(2). The amount of H(2)O(2) produced is greater at pH 5.5, the optimum pH for PPO activity, than at pH 4.5. Hence, an observed increase of theaflavins in black teas fermented at pH 4.5 appears to be due to lower turnover of formed theaflavins into thearubigins.     

Metabolic dependence of green tea on plucking positions revisited: a metabolomic study

The dependence of global green tea metabolome on plucking positions was investigated through (1)H nuclear magnetic resonance (NMR) analysis coupled with multivariate statistical data set. Pattern recognition methods, such as principal component analysis (PCA) and orthogonal projection on latent structure-discriminant analysis (OPLS-DA), were employed for a finding metabolic discrimination among fresh green tea leaves plucked at different positions from young to old leaves. In addition to clear metabolic discrimination among green tea leaves, elevations in theanine, caffeine, and gallic acid levels but reductions in catechins, such as epicatechin (EC), epigallocatechin (EGC), epicatechin-3-gallate (ECG), and epigallocatechin-3-gallate (EGCG), glucose, and sucrose levels were observed, as the green tea plant grows up. On the other hand, the younger the green tea leaf is, the more theanine, caffeine, and gallic acid but the lesser catechins accumlated in the green tea leaf, revealing a reverse assocation between theanine and catechins levels due to incorporaton of theanine into catechins with growing up green tea plant. Moreover, as compared to the tea leaf, the observation of marked high levels of theanine and low levels of catechins in green tea stems exhibited a distinct tea plant metabolism between the tea leaf and the stem. This metabolomic approach highlights taking insight to global metabolic dependence of green tea leaf on plucking position, thereby providing distinct information on green tea production with specific tea quality.     

Modulation of cholesterol metabolism by the green tea polyphenol (-)-epigallocatechin gallate in cultured human liver (HepG2) cells

Epidemiological and animal studies have found that green tea is associated with lower plasma cholesterol. This study aimed to further elucidate how green tea modulates cholesterol metabolism. When HepG2 cells were incubated with the main green tea constituents, the catechins, epigallocatechin gallate (EGCG) was the only catechin to increase LDL receptor binding activity (3-fold) and protein (2.5-fold) above controls. EGCG increased the conversion of sterol regulatory element binding protein-1 (SREBP-1) to its active form (+56%) and lowered the cellular cholesterol concentration (-28%). At 50 microM, EGCG significantly lowered cellular cholesterol synthesis, explaining the reduction in cellular cholesterol. At 200 microM EGCG, cholesterol synthesis was significantly increased even though cellular cholesterol was lower, but there was a significant increase seen in medium cholesterol. This indicates that, at 200 microM, EGCG increases cellular cholesterol efflux. This study provides mechanisms by which green tea modulates cholesterol metabolism and indicates that EGCG might be its active constituent.     

Adsorption behavior of the catechins and caffeine onto polyvinylpolypyrrolidone

Adsorbent is one of the most important factors for separation efficiency in fixed-bed purification techniques. The adsorption behavior of catechins and caffeine onto polyvinylpolypyrrolidone (PVPP) was investigated by static adsorption tests. The results showed that catechins rather than caffeine were preferred to adsorb onto PVPP since the adsorption selectivity coefficient of total catechins vs caffeine was around 22.5, and that adsorption of catechins could be described by the pseudo-second-order model. Adsorption amount of caffeine onto PVPP in green tea extracts solution was much higher than that in purified caffeine solution although the initial concentration of caffeine was similar in the two solutions, indicating the caffeine might be attached with catechins which were adsorbed by PVPP instead of being adsorbed by PVPP directly. The results also showed that the adsorption capacity of catechins and caffeine decreased with an increase in temperature, and that Freundlich and Langmuir models were both suitable for describing the isothermal adsorption of catechins, but not suitable for caffeine. The predicted maximum monolayer adsorption capacity of total catechins by PVPP was 671.77 mg g(-1) at 20 °C, which was significantly higher than that by other reported adsorbents. The thermodynamics analyses indicated that the adsorption of catechins onto PVPP was a spontaneous and exothermic physisorption process, revealing lower temperature was favorable for the adsorption of catechins. Elution tests showed that the desorption rates of catechins and caffeine were higher than 91% and 99% after two elution stages; in detail, almost all of the caffeine could be washed down at the water eluting stage, while catechins could be recovered at the dimethyl sulfoxide/ethanol solution eluting stage. Thus, the PVPP could be used as an excellent alternative adsorbent candidate for separating catechins from crude tea extracts, although some investigations, such as exploring the new eluants with low boiling point and high desorption efficiency, should be conducted furthermore.     

Tea catechin supplementation increases antioxidant capacity and prevents phospholipid hydroperoxidation in plasma of humans.

The effect of green tea catechin supplementation on antioxidant capacity of human plasma was investigated. Eighteen healthy male volunteers who orally ingested green tea extract (254 mg of total catechins/subject) showed 267 pmol of epigallocatechin-3-gallate (EGCg) per milliliter of plasma at 60 min after administration. The plasma phosphatidylcholine hydroperoxide (PCOOH) levels attenuated from 73.7 pmol/mL in the control to 44.6 pmol/mL in catechin-treated subjects, being correlated inversely with the increase in plasma EGCg level. The results suggested that drinking green tea contributes to prevent cardiovascular disease by increasing plasma antioxidant capacity in humans.     

Determination of tea polyphenols and caffeine in tea flowers (Camellia sinensis) and their hydroxyl radical scavenging and nitric oxide suppressing effects

The native occurrence of tea polyphenols, namely, (-)-epicatechin, (+)-catechin, (-)-epigallocatechin 3-gallate, (-)-epicatechin, and (-)-epicatechin 3-gallate, and caffeine in tea flowers was assessed by an isocratic HPLC procedure. The levels of total catechins and caffeine were determined in tea flowers collected from 10 different species of Camellia sinensis. The results showed the levels of total catechin ranged from 10 to 38 mg/g, whereas the level of caffeine ranged from 3 to 8 mg/g. Levels of catechins and caffeine in tea leaves and various teas were also determined and ranged from 2 to 126 mg/g and from 23 to 49 mg/g, respectively. Both tea flower and tea leaf extracts exert their strong hydroxyl radical scavenging effects in the Fenton reaction system and nitric oxide suppressing effects in LPS-induced RAW 264.7 cells. Most tea flowers contain less caffeine, but comparable amounts of total catechins, compared to tea leaves and teas. The present study demonstrates that both tea flowers and tea leaves contain appreciable amounts of catechins and caffeine. It is likely that tea flowers might be useful for making alternative tea beverages.     

Radical scavenging conserves from unused fresh green tea leaves

Green teas were made by inactivating the enzymes present in fresh leaves of coarse/pruned (unused) and normal (used for tea) grades using different sources of thermal energies. Green teas were extracted in a Soxhlet using different solvents. The obtained miscella was subjected to concentration to give the extract. The extract was subjected to solvent-solvent extraction. Solvent extract was concentrated to obtain conserve. The yields of conserves are 17 +/- 0.8 and 15 +/- 0.8% from green teas of normal and coarse tea leaves, respectively. The radical scavenging activity of these extracts was evaluated using a DPPH in vitro model system. The total polyphenol content was also determined and found to be higher in conserves from normal tea leaves. However, radical scavenging activity of conserves from coarse and normal green tea leaves was found to be >90% at 15 ppm concentration. The HPLC profiles of these conserves were used to quantify the total catechin content with the help of calibration curves prepared using authentic samples at known concentrations. The total catechin content is found to be in the range of 55-85%. Results indicated that the extracts from coarse leaves also possess potential biological activity and could be used as nutraceuticals as well as for preservation purposes in food formulations.     

外源NO对采后绿芦笋木质化和抗氧化能力的影响

为探讨NO处理对采后绿芦笋木质化合成的调控机理。以0.2mmol/L的硝普钠为一氧化氮供体,以蒸馏水处理为对照,研究了NO对采后绿芦笋总酚、木质素含量;木质素合成关键酶(苯丙氨酸解氨酶、肉桂醇脱氢酶、过氧化物酶、多酚氧化酶);抗氧化能力(1,1-二苯基-2-三硝基苯肼自由基清除率、超氧阴离子自由基(O2.-)清除率、羟基自由基(·OH)清除率)及膜透性的影响,结果表明,与对照相比,NO处理可延缓采后绿芦笋木质素含量和膜透性的增加,抑制肉桂醇脱氢酶和多酚氧化酶活性,增强抗氧化能力,诱导过氧化物酶活性增强,但对苯丙氨酸解氨酶活性和总酚含量的影响不显著。说明NO处理是通过抑制肉桂醇脱氢酶和多酚氧化酶活性、提高抗氧化能力而延缓采后绿芦笋的木质化进程。研究结果可为NO在绿芦笋保鲜中的应用提供理论依据和技术方法。