Effects of catechin concentration on the formation of black tea polyphenols during in vitro oxidation

An in vitro model fermentation system, containing purified catechins and partially purified polyphenol oxidase (EC 1.14.81.1) from green tea shoots, has been used to determine the efrect of catechin mixtures of different concentration and proportions on the formation of theaflavin and thearubigin. Increases in total catechin concentration, 25% above that typical in green tea shoots of Malawi-grown bushes, inhibited polyphenol oxidase activity and, consequently, depressed theaflavin levels. Individual or combined concentrations of epicatechin gallate and epigallocatechin gallate in excess of 110 mM were shown to be responsible for enzyme inhibition, whereas epicatechin and epigallocatechin had no effect. Fermentation of a catechin mixture, containing the four major catechins, epicatechin, epigallocatechin, epigallocatechin gallate and epicatechin gallate, at equal individual concentrations (55 mM), produced, after 3O min, total theaflavin levels 68% higher and thearubigin levels only 25% higher than those from a standard catechin mixture fermented under similar conditions. Continued fermentation of this mixture produced no further theaflavin, but the thearubigin fraction increased significantly, due to subsequent oxidation of the excess of simple catechins. A new catechin mixture was, therefore, calculated to give a similar level of theaflavin to that of the previous mixture without leaving an excess of unoxidized simple catechins. The catechin proportions and concentrations of the latter mixture agree well with those of the green shoots of quality Kenyan teas or similar quality Malawi teas grown during the dry cold season. The results indicate that a high ratio of simple to gallocatechins will facilitate a high theaflavin-thearubigin ratio in black tea.     

Determination of catechins in human urine subsequent to tea ingestion by high-performance liquid chromatography with electrochemical detection

The title determination was conducted by HPLC with electrochemical detection using an ODS column and a mobile phase of acetonitrile: 0.1 M phosphate buffer (pH 2.5) (15:85, v/v). The eight catechins, gallocatechin (GC), epigallocatechin (EGC), catechin (C), epicatechin (EC), epigallocatechin gallate (EGCg), gallocatechin gallate (GCg), epicatechin gallate (ECg), and catechin gallate (Cg), were detected at 0.6 V vs Ag/AgCl. Good linear relationships between current and amount were noted for 0.5-250 pmol of each catechin, with a correlation coefficient of 0.999 in each case. The detection limit for any one was 0.5 pmol (signal to noise ratio, S/N = 3). After the ingestion of 340 ml canned green tea, GC, EGC, C, and EC, mostly in conjugated form, were determined in urine samples. Conjugated catechins were hydrolyzed by enzymes using sulfatase and beta-glucuronidase. The time courses of the above four catechins showed a maxima at 1-3 h after tea ingestion. (+), (-)-EC and (+), (-)-C were present in canned tea.     

Recovery of catechin compounds from Korean tea by solvent extraction

Catechin compounds from Korean green tea as potential sources of anticancer and antioxidant components were target materials in this work. The methodologies of solvent extraction and partition were utilized to recover catechin compounds from green tea. The optimum experimental condition was obtained by optimizing operating factors, such as, the extraction solvent, extraction time and operating temperature. After extracting the green tea with water at 80 degrees C for 40 min, the extract was partitioned with water/chloroform, which was best suited to remove caffeine impurity from the extract. Further, the resulting extract was partitioned water/ethyl acetate to deeply purify the catechin compounds of EGC, EC, EGCG and ECG. The experimental result in this work could be extended to preparative HPLC to obtain EGCG on commercial scale.     

Enhancement of phagocytic activity of macrophage-like cells by pyrogallol-type green tea polyphenols through caspase signaling pathways

We investigated the phagocytosis-enhancing activity of green tea polyphenols, such as epigallocatechin gallate (EGCG), epigallocatechin (EGC), epicatechin gallate (ECG), epicatechin (EC) catechin (+C) and strictinin, using VD3-differentiated HL60 cells. EGCG, EGC, ECG and strictinin, but not EC and +C, increased the phagocytic activity of macrophage-like cells, and a caspase inhibitor significantly inhibited phagocytic activities. These results suggest that the pyrogallol-type structure in green tea polyphenols may be important for enhancement of the phagocytic activity through caspase signaling pathways.     

白化和黄化茶树品种绿茶游离氨基酸、儿茶素类及咖啡碱差异分析

为探究以白化和黄化茶树品种鲜叶为原料制成的绿茶滋味品质和代谢物差异,对广德市6个白化品种绿茶(奶白茶)和14个黄化品种绿茶(黄金芽茶)进行感官审评和代谢物分析。结果表明,奶白茶滋味鲜爽而收敛性略弱;黄金芽茶滋味收敛性强而鲜度低于奶白茶。游离氨基酸总量以及呈现鲜味、甜味的游离氨基酸在奶白茶中的含量显著高于黄金芽茶,而贡献收敛性的儿茶素类化合物和没食子酸含量以及呈现苦味的咖啡碱含量在奶白茶中显著低于黄金芽茶。偏最小二乘法判别分析(PLS-DA)表明导致两种绿茶滋味差异的标志性化合物有7种,分别是茶氨酸、精氨酸、谷氨酸、天冬氨酸、表没食子儿茶素没食子酸酯(EGCG)、表儿茶素没食子酸酯和咖啡碱。味觉活性值(Dot)最高的EGCG在黄金芽茶中的呈味贡献显著高于奶白茶。因此,游离氨基酸、儿茶素类化合物、没食子酸和咖啡碱含量差异导致白化和黄化茶树品种绿茶滋味不同。     

Epigallocatechin gallate and gallocatechin gallate in green tea catechins inhibit extracellular release of Vero toxin from enterohemorrhagic Escherichia coli O157:H7

We studied the effects of six catechin derivatives (catechin, epigallocatechin, epicatechin, epicatechin gallate, epigallocatechin gallate (EGCg) and gallocatechin gallate (GCg)) in green tea on the production and extracellular release of Vero toxins (VTs) from enterohemorrhagic Escherichia coli (EHEC) cultured at 37 degrees C for 24 h. EGCg and GCg in the culture medium markedly inhibited extracellular VTs release from EHEC cells into the culture supernatant fluid at concentrations of 0.05 mg/ml or higher, as estimated by both the reversed passive latex agglutination assay and cytotoxic assay using Vero cells. Production and extracellular release of maltose binding protein, a periplasmic protein, into the culture supernatant were also inhibited by EGCg and GCg, indicating that their inhibitory effect on release from periplasm into the outer milieu is not specific to VTs, but general to the proteins accumulated in EHEC periplasm.     

Preparative isolation and purification of theaflavins and catechins by high-speed countercurrent chromatography

High-speed countercurrent chromatography (HSCCC) has been applied for the separation of theaflavins and catechins. The HSCCC run was carried out with a two-phase solvent system composed of hexane-ethyl acetate-methanol-water-acetic acid (1:5:1:5:0.25, v/v) by eluting the lower aqueous phase at 2 ml/min at 700 rpm. The results indicated that pure theaflavin, theaflavins-3-gallate, theaflavins-3'-gallate and theaflavin-3,3'-digallate could be obtained from crude theaflavins sample and black tea. The structures of the isolated compounds were positively confirmed by (1)H NMR and (13)C NMR, MS analysis, HPLC data and TLC data. Meanwhile, catechins including epigallocatechin gallate, gallocatechin gallate, epicatechin gallate and epigallocatechin were isolated from the aqueous extract of green tea by using the same solvent system. This study developed a modified method combined with enrichment theaflavins method by using HSCCC for separation of four individual theaflavins, especially for better separation of theaflavins monogallates.     

Keemun black tea extract contains potent fatty acid synthase inhibitors and reduces food intake and body weight of rats via oral administration

The inhibitory effects of a black tea extract on fatty acid synthase were measured through inhibition kinetics. The Keemun black tea extract showed more potent inhibitory activity on fatty acid synthase than green tea extract. Additionally, the inhibitory ability of the black tea extract depended on the extracting solvent and the conditions used. Only 10-23% of the inhibitory activity from the black tea was extracted by the general method of boiling with water. The results suggested that the main fatty acid synthase inhibitors in black tea might be theaflavins. Which were more potent than epigallocatechin gallate or C75. The reaction site on the fatty acid synthase and the inhibition kinetics for the extract were different from those of epigallocatechin gallate or C75. In addition, Keemun black tea extract significantly reduced food intake, body weight and blood triglyceride of diet-induced obesity SD rats via oral administration.     

Adsorption affinity of tea catechins onto polymeric resins: Interpretation from molecular orbital theory

Adsorption of certain tea catechins such as (+) catechin (C), (−) epicatechin (EC), (+) gallocatechin (GC), (−) epigallocatechin (EGC), (+) catechin gallate (CG), (−) epicatechin gallate (ECG), (+) gallocatechin gallate (GCG), and (−) epigallocatechin gallate (EGCG) have been studied using three types of polymeric resins as adsorbents. Adsorption affinity expressed as the slope of the linear region of the isotherm for a solute is found to be different for different adsorbents, and this difference can be interpreted from the chemical nature of the sorbents. Molecular interactions on polymeric resins have been studied based on molecular orbital theory. Electronic states of adsorbent and adsorbate were calculated using the semiempirical molecular orbital (MO) method from which energy of adsorption in aqueous solution was estimated. The adsorptive interaction on the polymeric resins computed on the basis of frontier orbital theory seems to correlate well with the experimentally measured adsorption affinity and enthalpy.     

Complex effects of different green tea catechins on human platelets

Epigallocatechin gallate (EGCG), a major component of green tea, has been previously shown to inhibit platelet aggregation. The effects of other green tea catechins on platelet function are not known. Pre-incubation with EGCG concentration-dependently inhibited thrombin-induced aggregation and phosphorylation of p38 mitogen-activated protein kinase and extracellular signal-regulated kinases-1/2. In contrast EGCG stimulated tyrosine phosphorylation of platelet proteins, including Syk and SLP-76 but inhibited phosphorylation of focal adhesion kinase. Other catechins did not inhibit platelet aggregation. Interestingly, when EGCG was added to stirred platelets, a tyrosine kinase-dependent stimulation of platelet aggregation was observed. The two other catechins containing a galloyl group in the 3' position (catechin gallate, epicatechin gallate) also stimulated platelet aggregation, while catechins without a galloyl group (catechin, epicatechin) or the catechin with a galloyl group in the 2' position (epigallocatechin) did not.     

Interaction of Tea Catechins with Lipid Bilayers Investigated with Liposome Systems

Interaction of tea catechins with lipid bilayers was investigated with liposome systems, which enabled us to separate liposomes from the external medium by centrifugation. We found that epicatechin gallate had the highest affinity for lipid bilayers, followed by epigallocatechin gallate, epicatechin, and epigallocatechin. Epicatechin gallate and epigallocatechin gallate in the surface of lipid bilayer perturbed the membrane structure.     

人工接种冠突散囊菌对白茶主要呈味物质的影响

本文为了排除其他微生物的干扰,首次以人工接种的方式研究了冠突散囊菌Eurotium cristatum对白茶主要呈味物质的作用。采用高效液相色谱、分光光度计等方法,分析表明冠突散囊菌能够显著降低白茶中呈苦涩味的表没食子儿茶素没食子酸酯（EGCG）和表儿茶素没食子酸酯（ECG）,并提高表没食子儿茶素（EGC）、Asp、His、咖啡碱和山奈酚的含量;灭菌压制的过程中EGCG可能异构化成为更稳定的没食子儿茶素没食子酸酯（GCG）,且二者以相近的含量共存。冠突散囊菌可以降低人工发花白茶饼中呈苦涩味的化合物含量,从而达到减少白茶饼苦涩味的效果;灭菌压制过程也能够降低白茶饼的苦涩味物质的含量。“发花”处理为白茶带来了新的风味,可以丰富白茶产品种类,同时为促进粗老原料白茶的综合利用提供新思路。     

Interaction of tea catechins with lipid bilayers investigated with liposome systems

Interaction of tea catechins with lipid bilayers was investigated with liposome systems, which enabled us to separate liposomes from the external medium by centrifugation. We found that epicatechin gallate had the highest affinity for lipid bilayers, followed by epigallocatechin gallate, epicatechin, and epigallocatechin. Epicatechin gallate and epigallocatechin gallate in the surface of lipid bilayer perturbed the membrane structure.     

Effects of physical and chemical conditions on the in vitro oxidation of tea leaf catechins

The use of a model fermentation system containing purified green tea shoot catechins and partially purified polyphenol oxidase (EC 1.14.18.1) has provided important data on theaflavin and thearubigin formation. Low oxygen concentration during fermentation, as a result of inadequate aeration or high enzyme concentration, and enhanced by high temperatures, inhibits theaflavin and promotes thearubigin production. Analysis of catechin oxidation under low oxygen tension suggests that the quinones of epicatechin and epicatechin gallate, by virtue of their low redox potentials, are acting as electron carriers in the preferential oxidation of epigallocatechin, epigallocatechin gallate and theaflavin intermediates. Temperature changes between 20° and 30° had little effect on theaflavin values, whereas thearubigin production increased dramatically. Thearubigin-theaflavin ratios over this temperature range increased in oxygen from 1.5:1 to 2.5:1. The pH optima for theaflavin and thearubigin formation were 5 and 6, respectively. The optimization of this factor, together with oxygen concentration, temperature and enzyme concentration, for theaflavin formation, resulted in theaflavin levels 115% higher than those obtained from similar model fermentations occurring under the conditions associated with black tea manufacture in Malawi (pH 5.6; 30 × 10^?8 kat of polyphenol oxidase; 27°; 20% oxygen).     

Dietary green and white teas suppress UDP-glucuronosyltransferase UGT2B17 mediated testosterone glucuronidation

The anabolic steroid testosterone can be used by athletes to enhance athletic performance and muscle growth. UDP-glucuronosyltransferase (UGT2B17) is the key enzyme involved in the glucuronidation of testosterone to testosterone glucuronide, which also serves as a marker for the testosterone/epitestosterone (T/E) ratio used to detect testosterone abuse in sport. Inhibitors of testosterone glucuronidation could have an impact on circulating testosterone levels, thus aiding performance, as well as potentially affecting the urinary T/E ratio and therefore masking testosterone abuse. Previous reports have revealed that non-steroidal, anti-inflammatory drugs, diclofenac and ibuprofen, inhibit the UGT2B17 enzyme. The aim of this study is to analyse dietary tea samples for inhibition of testosterone glucuronidation and, where inhibition is present, to identify the active compounds. Analysis of testosterone glucuronidation was conducted by performing UGT2B17 assays with detection of un-glucuronidated testosterone using high performance liquid chromatography. The results from this study showed that testosterone glucuronidation was inhibited by the green and white tea extracts, along with specific catechin compounds, notably: epicatechin, epigallocatechin gallate (EGCG) and catechin gallate. The IC50 inhibition value for EGCG was determined, using a Dixon plot, to be 64 μM, equalling the most active NSAID inhibitor diclofenac. Thus, common foodstuffs and their constituents, for the first time, have been identified as inhibitors of a key enzyme involved in testosterone glucuronidation. Whilst these common compounds are not substrates of the UGT2B17 enzyme, we showed that they inhibit testosterone glucuronidation which may have implications on current doping control in sport.     

Relationship between the anti-hemolysin activity and the structure of catechins and theaflavins

We examined the corresponding isomers of catechins and theaflavins for anti-hemolysin activities against Staphylococcus aureus alpha-toxin and Vibrio cholerae O1 hemolysin. Catechins and theaflavins showed anti-hemolysin activities in a dose-dependent manner. Among the catechins tested, (-)catechin gallate, (-)epicatechin gallate and (-)epigallocatechin gallate having galloyl groups in their molecules showed more potent anti-hemolysin activities against both toxins. On the other hand, free catechins, i. e. (-)catechin, (-)gallocatechin, (-) epicatechin and (-)epigallocatechin had low anti-hemolysin activities against alpha-toxin. Although (-)catechin or (-)gallocatechin had no effect on cholera hemolysin, (-) epicatechin and (-)epigallocatechin were slightly inhibitory. Among dextrocatechins, (+) epicatechin and (+)epigallocatechin proved to be more effective than (+)catechin and (+) gallocatechin. The anti-hemolysin activities of theaflavins against alpha-toxin and cholera hemolysin were dependent on the number of the galloyl group in their structure. These results suggest that the tertiary structure of the catechin or theaflavin and the active site of hemolysin, that affects the interaction between them, plays an important role in the anti-hemolysin activity.     

A shortcut from plasma to chromatographic analysis: Straightforward and fast sample preparation for analysis of green tea catechins in human plasma

This paper describes a new and straightforward method for determination of the green tea catechins epicatechin, epicatechin gallate, epigallocatechin, and epigallocatechin gallate in human plasma. Sample preparation includes addition only of dimethylformamide and trichloroacetic acid. After centrifugation, the supernatant can be injected into the HPLC. If required, the glucuronides and sulphates of the catechins can be enzymatically hydrolysed before extraction. Recovery ranges from 92.9 to 98.2%; limits of detection, from 2.4 to 5.0 ng/mL; and relative standard deviations, from 3.1 to 8.6%. Twelve samples can be processed within 45 min, and are then ready to be injected into the HPLC. The method was successfully applied to human plasma. This method is suitable for studies on absorption, bioavailability, and kinetics of green tea catechins in plasma. Since manual work and time consumption are minimal, the procedure is especially useful for large numbers of samples.     

An analytical survey of the polyphenols of seeds of varieties of grape (Vitis vinifera) cultivated in Greece: implications for exploitation as a source of value-added phytochemicals

Seed samples from 12 white and 25 red international and Hellenic native grape varieties (Vitis vinifera) were screened for their polyphenolic composition. The polyphenols determined were mainly of low molecular weight, including gallic acid, catechin, epicatechin, epicatechin gallate, epigallocatechin, epigallocatechin gallate and the procyanidins B1 and B2. Average values of total content for white and red varieties (376 and 388 mg/100 g seeds, respectively) were very similar. Comparable results were observed with respect to the individual polyphenol content with seeds from red varieties being, in general, slightly richer. The predominant flavanol monomer in white and red varieties was catechin (which accounted for 50.5 and 49.3%, respectively, of the total content), whilst gallic acid and epigallocatechin were the constituents showing the lowest content, respectively. The data obtained are discussed with regard to the exploitation of grape seeds as a low-cost source of value-added phytochemicals.     

Affinity of polyphenols for lipid bilayers

Interaction of tea catechins with lipid bilayers has been investigated with liposome systems. Epicatechin gallate had the highest affinity for lipid bilayers, followed by epigallocatechin gallate, epicatechin, and epigallocatechin. Epicatechin gallate and epigallocatechin gallate in the surface of lipid bilayer perturbed the membrane structure.     

Interaction of tea catechins with lipid bilayers investigated by a quartz-crystal microbalance analysis

The quartz-crystal microbalance (QCM) technique was applied to investigate the interaction of tea catechins with lipid bilayers. The association constants obtained from the frequency changes of QCM revealed that (-)epicatechin gallate and (-)epigallocatechin gallate interacted with 1,2-dimyristoyl-sn-glycero-3-phosphocholine ca. 1000 times more strongly than (-)epicatechin and (-)epigallocatechin. The results exhibited good correlation with the strength of biological activity.