Catechin stability of EGC- and EGCG-enriched tea drinks produced by a two-step extraction procedure

Catechin content of green tea drinks commercially available is reported to be very low in comparison with tea traditionally prepared, due to catechins conversion to their corresponding epimers during production. The purpose of this present study was to produce catechin-enriched tea drinks according to a two-step brewing procedure and to verify the catechin stability of those enriched drinks during storage. Those results confirmed that it is possible to produce EGC- and EGCG-enriched tea drinks regardless of the green tea used. Good extraction efficiencies were reached for the first and second extraction steps with catechin extraction yields ranging from 63.6% to 84.8%. Furthermore, it appeared that the catechin content in the two enriched tea drinks demonstrated a great stability since no significant degradation occurred within 8 weeks of storage. This simple two-step extraction procedure could be considered as an interesting way to produce enriched green tea drinks with more potent and stable bioactive catechins.     

不同茶饮料贮藏过程中品质的变化

本文从色泽、浊度及活性成分含量三个方面分析不同茶饮料的品质变化特征，分别对高酸绿茶饮料、低酸绿茶饮料、高酸红茶饮料及低酸红茶饮料在不同贮藏条件下的可溶性固形物含量、透光率、浊度、色差、茶多酚含量、表没食子儿茶素没食子酸酯（Epigallocatechin gallate，EGCG）含量等指标进行测定，探究不同茶饮料在不同贮藏条件（温度、时间）下品质变化的情况。结果表明:不同茶饮料在贮藏过程中，透光率无显著变化；高酸绿茶饮料在4 ℃及25 ℃条件下贮藏，随贮藏时间的延长，浊度显著增加，贮藏至第180 d，分别增加了52.04%、34.12%，红茶饮料浊度无显著变化；不同茶饮料在贮藏过程中，随贮藏时间的延长，L*值降低，a*值及b*值增加，25 ℃条件下贮藏至180 d，高酸绿茶饮料、低酸绿茶饮料、高酸红茶饮料及低酸红茶饮料的L*值分别降低了31.86%、39.98%、30.22%及37.73%；茶饮料中的茶多酚及EGCG含量随着贮藏时间的延长而降低，EGCG降解程度较茶多酚高，在55 ℃条件下贮藏15 d，高酸绿茶饮料、低酸绿茶饮料、高酸红茶饮料及低酸红茶饮料的茶多酚含量分别降低9.08%、10.28%、13.39%及18.63%，EGCG含量分别下降了35.32%、42.18%、47.22%及49.03%。     

Digestive stability and absorption of green tea polyphenols: Influence of acid and xylitol addition

The objective of this study was to determine the effect of sugar substitute and acid on digestive stability and intestinal uptake of catechins in Ready-To-Drink (RTD) green tea. Green tea extracts formulated with prescribed amounts of sucrose (200, 500, and 1000 mg), glucose (280, 700, and 1400 mg), and xylitol (200, 500, and 1000 mg) in combination with citric acid (10 mg) or vitamin C (10 mg) were subjected to an in vitro digestion model coupled with Caco-2 cells. Green tea extracts only showed a poor digestive recovery (5.3%) of total catechins and EGC and EGCG significantly decreased with the digestive recovery of 4.6% and 6.1%, respectively. However, measured amount of EGC, EGCG, or ECG in digestive fluids and caco-2 human intestinal cell significantly increased by adding citric acid or vitamin C. There was remarkable increase of digestive recovery of total catechins in green tea with xylitol/citric acid and xylitol/vitamin C by 1.7–2.5 times and 3 times, respectively, with different amounts of xylitol. It was also determined that intestinal uptake of total catechins significantly increased 6 and 11 times in green tea with xylitol/citric acid and xylitol/vitamin C, respectively, compared to green tea only.     

Color and chemical stability of tea polyphenol (−)-epigallocatechin-3-gallate in solution and solid states

(−)-Epigallocatechin-3-gallate (EGCG), the main component with the highest biological activity in green tea, has been extensively studied for its activities and degradation behaviors in infusions and solutions. However, neither the characterization of its degradation products in solutions, nor its color degradation and stability in the solid state are well documented. Therefore, in this present study, the color and chemical stability of EGCG in solutions with various concentrations ranging from 0.05 mg/mL to 30 mg/mL and in the solid state following storage at 0% and 85% relative humidity (RH), 80 °C were investigated. Color parameters were monitored over time, and the chemical stability was determined using high performance liquid chromatography (HPLC)–diode array detection (DAD) analysis and HPLC–mass spectrometry (MS). Mathematical models for color degradation of EGCG in both solution and solid states were established. The chemical stability of EGCG was found to be affected by its physical state, concentration of reactants, oxygen levels, pH, and RH conditions. Epimerization and oxidation were found to be the two major reactions causing the degradation of EGCG in the solution and solid state, respectively. Major degradation products, catechin monomers and oxidation dimers, were identified. Cleavage products, gallic acid and catechin monomers, were also found. These results further elaborated the degradation mechanisms of EGCG in solutions of varying concentrations and revealed the chemical changes resulting in color degradation of EGCG in the solid state.     

Albumin causes a synergistic increase in the antioxidant activity of green tea catechins in oil-in-water emulsions

Model oil-in-water emulsions containing epicatechin (EC) and epigallocatechin gallate (EGCG) showed a synergistic increase in stability in emulsions containing added albumin. EGCG showed a stronger synergy (35%) with ovalbumin than did EC. Oxidation of the oil was monitored by determining peroxide values and hexanal contents. The effect of bovine serum albumin (BSA) on model oil-in-water emulsions containing each of the green tea catechins [epicatechin gallate (ECG), EGCG, EC and epigallocatechin (EGC)] was studied during storage at 30 °C. The green tea catechins showed moderate antioxidant activity in the emulsions with the order of activity being ECG ≈ EGCG > EC > EGC. Although BSA had very little antioxidant activity in the absence of phenolic antioxidants, the combination of BSA with each of the catechins showed strong antioxidant activity. BSA, in combination with EC, EGCG or EGC, showing the strongest antioxidant activity with good stability after 45 days storage. Model experiments with the catechins stored with BSA in aqueous solutions confirmed that protein–catechin adducts with antioxidant activity were formed between the catechins and protein. The antioxidant activity of the separated protein–catechin adducts increased strongly with storage time and was stronger for EGCG and ECG than for EC or EGC.     

Milk enhances intestinal absorption of green tea catechins in in vitro digestion/Caco-2 cells model

The effect of milk on the absorption of polyphenols is still controversial so far. In order to determine the impact of milk addition on green tea catechins bioaccessibility and intestinal absorption an in vitro digestion/Caco-2 cell model was applied. Green tea extract (GTE) was solubilized in distilled water at 23 °C and 100 °C, combined with skimmed milk (GTE + 10% milk and GTE + 25% milk) and subjected to simulated gastric and intestinal digestion, followed by transepithelial absorption in Caco-2 cells monolayers. In the mixture with milk, gallated catechins: ECG and EGCG showed binding to milk proteins while EC and EGC seemed to have weaker affinity. Catechins were stable during gastric incubation and very sensitive to intestinal digestion. Bioaccessibility of green tea catechins brewed at 100 °C was higher than brewed at 23 °C. Catechins from digested GTE with 10% and 25% milk exhibited enhanced intestinal permeability in Caco-2 model in comparison to non-digested GTE and digested GTE without milk. Apparent permeability coefficients (P_app) of EGCG and ECG in digested GTE with 25% milk were significantly higher compared to those in GTE with 10% milk, and amounted to 2.41 × 10^− 6 cm/s and 1.39 × 10^− 6 cm/s. The recoveries of all catechins in GTE with milk in Caco-2 cells after 2 h incubation were significantly higher than that without milk. To summarize, these data suggest that milk addition may increase catechin bioavailability by enhancing their transepithelial absorption and uptake from green tea extract.     

Investigation of the interaction between soluble antioxidants in green tea and insoluble dietary fiber bound antioxidants

This work investigates the possibility of interaction between insoluble dietary fiber bound antioxidants, specifically of wheat bran, and soluble antioxidants like those provide by aqueous infusions of green tea. Solutions of pure catechins were also assayed for comparison with those naturally found in tea. To accomplish this, the aqueous and alcohol soluble fractions as well as the lipid components of wheat bran were firstly removed and the freeze-dried insoluble residue was then treated with different concentrations of green tea infusions or aqueous solutions of epicatechin (EC) and epigallocatechin-3-gallate (EGCG) for certain time. Treatment with EC (0–200 μM) had no significant effect on the antioxidant capacity of insoluble bran fraction. However, treatment with EGCG significantly (p < 0.05) increased linearly the antioxidant capacity as a function of concentration (0–100 μM). Treatment with great tea infusions (1–3 g/100 ml) also increased the resulting antioxidant capacity of insoluble bran fraction, but the effect was lesser at higher infusion concentrations. Liquid chromatography couple to mass spectrometry (LC–MS) analyses of aqueous phases after treatment indicated comparable levels of decrease in the concentrations of catechins confirming their reaction with the radical forms of antioxidants bound to insoluble bran matrix.     

Total polyphenols,catechin profiles and antioxidant activity of tea products from purple leaf coloured tea cultivars

Black (aerated) and green (unaerated) tea products, processed from 10 green and 18 purple leaf coloured cultivars of Kenyan origin, and two tea products, from the Japanese cultivars, Yabukita and Yutakamidori, were assayed for total polyphenols (TP) content, individual catechin profiles and in vitro antioxidant capacity (AA). In addition, the phenolic content of the tea products was determined using the Folin–Ciocalteu phenol reagent. Catechin fractions were identified using reverse phase high performance liquid chromatography (HPLC) with a binary gradient elution system.The AA% of the tea products was determined using a 2,2′-diphenyl picrylhydrazyl (DPPH) radical assay method. The results showed that TPs, catechin profiles and antioxidant activities were significantly (p ? 0.05) higher in unaerated than in aerated teas. Tea products from the purple leaf coloured tea cultivars had levels of TPs, total catechin (TC) and antioxidant activities similar to those from the green leaf coloured cultivars, except for teas from the Japanese cultivars that were very low in the assayed parameters. Caffeine content was significantly (p ? 0.05) lower in products from the purple leaf coloured cultivars than in those from the green leaf coloured tea cultivars. Antioxidant activity (%) was higher in tea products from the Kenyan germplasm than in those from the Japanese cultivars. Antioxidant potency of tea products was significantly (r = 0.789⁷, p ? 0.01) influenced by the total anthocyanin content of the purple leaf coloured cultivars. Cyanidin-3-O-glucoside was the anthocyanin most highly correlated with AA% (r = 0.843⁷, p ? 0.01 in unaerated tea). Total catechins in the unaerated products from the green leaf coloured tea cultivars were also significantly correlated with antioxidant capacity (r = 0.818⁷, p ? 0.01). Results from this study suggest that the antioxidant potency of teas is dependent on the predominant flavonoid compound, the type of tea cultivar and the processing method.     

Effects of interval length between tasting sessions and sweetener level on long-term acceptability of novel green tea drinks

The objective of this study was to investigate the effects of the interval length between sessions (1-day vs. 3-day), the sweetener level (Optimal (OP) vs. Low (LO)), and the number of stimuli (one vs. two) on long-term acceptability of a novel green tea drink. Two consumer groups (N = 80) rated their liking of two drinks (OP and LO) while another 4 consumer groups (N = 162) rated only one drink over 10 sessions at either 1-day or 3-day intervals. Repeated consumption led to an overall increase in liking of the novel drink even though the patterns of increase were different according to the sweetener level and the interval length. Higher sweetener level and shorter interval length might contribute to increase in liking score during the earlier sessions and stable liking during the later sessions for novel green tea drink.     

Effects of aqueous brewing solution pH on the extraction of the major green tea constituents

The pH of the aqueous brewing solution was maintained at values ranging from 1 to 9 during the green tea extraction and the effects on the tea's extracted constituents were studied. The epistructured catechins were stable under acidic conditions but epimerized or degraded at pH ≥ 6. The extractable solids contained more epistructured catechins at pHs 3–5 but more non-epistructured catechins at pHs 6–7. More tea cream was obtained at pH 1 but the concentration of catechins, caffeine and theanine was low in this fraction. Therefore, to maximize the extraction of the epistructured catechins and to minimize their epimerization and degradation and to maximize the extraction of caffeine and theanine, the results suggest that the pH should be maintained between 3 and 5.3 during the aqueous brewing process.     

Effects of catechin-enriched green tea beverage on visceral fat loss in adults with a high proportion of visceral fat: A double-blind,placebo-controlled,randomized trial

The effects of catechin-enriched green tea on Chinese adults with a high proportion of abdominal visceral fat were evaluated. Subjects (118) were randomly assigned to consume daily a beverage containing 609.3 mg catechins and 68.7 mg caffeine or a control beverage for 12 weeks. Abdominal fat area, body weight and composition were measured at week 0, week 8, and week 12. One hundred and four subjects completed the trial. Average visceral fat area, body weight, and body fat were reduced significantly by catechin-enriched green tea treatment but these effects were not seen in the control group with per-protocol sets analysis. The decrease at week 12 in the visceral fat area in the catechin group was greater than that in the control group (P = 0.04). Thus, consumption of the catechin-enriched green tea beverage for 12 weeks induced visceral fat loss in Chinese adults with a high proportion of abdominal visceral fat.     

Green tea catechins during food processing and storage: A review on stability and detection

Green tea catechins can undergo degradation, oxidation, epimerization and polymerization during food processing. Many factors could contribute to the chemical changes of green tea catechins, such as temperature, pH of the system, oxygen availability, the presence of metal ions as well as the ingredients added. Several detection methods have been developed for tea catechin analysis, which are largely based on liquid chromatography (LC) and capillary electrophoresis (CE) methods for getting a good separation, identification and quantification of the catechins. Stability of green tea catechins is also influenced by storage conditions such as temperature and relative humidity. The stability of each catechin varies in different food systems and products. Pseudo first-order kinetic model has been developed and validated for the epimerization and degradation of tea catechins in several food systems, whereas the rate constant of reaction kinetics followed Arrhenius equation.     

Common tea formulations modulate in vitro digestive recovery of green tea catechins

Epidemiological evidence suggests a role for tea catechins in reduction of chronic disease risk. However, stability of catechins under digestive conditions is poorly understood. The objective of this study was to characterize the effect of common food additives on digestive recovery of tea catechins. Green tea water extracts were formulated in beverages providing 4.5, 18, 23, and 3.5 mg per 100 mL epicatechin (EC), epigallocatechin (EGC), epigallocatechin-gallate (EGCG), and epicatechin-gallate (ECG), respectively. Common commercial beverage additives; citric acid (CA), BHT, EDTA, ascorbic acid (AA), milk (bovine, soy, and rice), and citrus juice (orange, grapefruit, lemon, and lime) were formulated into finished tea beverages at incremental dosages. Samples were then subjected to in vitro digestion simulating gastric and small intestinal conditions with pre- and post-digestion catechin profiles assessed by HPLC. Catechin stability in green tea was poor with <20% total catechins remaining post-digestion. EGC and EGCG were most sensitive with less, not double equals 10% recovery. Teas formulated with 50% bovine, soy, and rice milk increased total catechin recovery significantly to 52, 55, and 69% respectively. Including 30 mg AA in 250 mL of tea beverage significantly (p<0.05) increased catechin recovery of EGC, EGCG, EC, and ECG to 74, 54, 82, and 45% respectively. Juice preparation resulted in the highest recovery of any formulation for EGC (81-98%), EGCG (56-76%), EC (86-95%), and ECG (30-55%). These data provide evidence that tea consumption practices and formulation factors likely impact catechin digestive recovery and may result in diverse physiological profiles.     

Inhibition of mackerel (Scomber scombrus) muscle lipoxygenase by green tea polyphenols

The high polyunsaturated fatty acid content of oily fish such as mackerel (Scomber scombrus) makes it particularly susceptible to oxidative degradation. We have shown previously the presence of lipoxygenase (LOX), a lipid oxygenase, in mackerel muscle. In the current study, commercially available green tea polyphenols were shown to effectively inhibit the LOX activity of mackerel muscle. EGCG (epigallocatechin gallate) was the strongest inhibitor tested with an IC₅₀ (concentration for half maximal inhibition) value of 0.13 nM. All the tea catechins showed a mixed non-competitive type inhibition. In addition, antioxidants such as BHA (butylated hydroxyanisole), BHT (butylated hydroxytoluene), esculetin, caffeic acid, ascorbic acid, and ethylene diamine tetraacetic acid (EDTA) were effective to varying degrees (IC₅₀ values between 0.02 and >50 μM) in the inhibition of mackerel muscle LOX. Nordihydroguaiaretic acid (NDGA), a classical LOX inhibitor and potassium cyanide (KCN), a heme protein inhibitor were assayed for their inhibitory activities for comparison.Post harvest spoilage of fish account for loss of as much as 10% of the world’s catches of cultured fish. This data indicates that the green tea polyphenols, nature’s very potent antioxidants, may be used as an effective and natural means of reducing post harvest spoilage in fish.     

Antioxidant packaging with encapsulated green tea for fresh minced meat

A novel approach to incorporate green tea extract into polyethylene by extrusion technology has been studied. Green tea entrapped by inorganic capsules was incorporated into melted material and extruded without compromising its quality. The shelf-life of fresh meat from two types of active packaging was significantly extended for 3 days compared to blank samples. Significant difference was observed after 9th day of study in case of results of CIE L*a*b*, MetMb and organoleptic assay. Migration study of antioxidants from the materials was also performed by UHPLC-MS/TQ using food simulants. Amount of migrated catechins was in the range between 6.3 ± 3.3 and 228.4 ± 15.2 μg/kg (ratio 6:1). Regression coefficients (R) between 0.9925 and 0.9989 were obtained. Minimum LOD (0.001 μg/g) and LOQ (0.004 μg/g) were obtained for epicatechin gallate and catechin gallate while maximum LOD (0.057 μg/g) and LOQ (0.191 μg/g) were obtained for catechin.     

Formulation with ascorbic acid and sucrose modulates catechin bioavailability from green tea

In order to investigate the impact of common food ingredients on catechin absorption, green tea (GT) extract (50 mg) was formulated plain, with sucrose (GT + S), with ascorbic acid (GT + AA) and with sucrose and ascorbic acid (GT + S + AA). Bioavailability and bioaccessibility were assessed in Sprague Dawley rats and an in vitro digestion/Caco-2 cell model respectively. Absorption of epigallocatechin (EGC) and epigallocatechin gallate (EGCG) was significantly (P < 0.05) enhanced in GT + S + AA formulations (AUC_0–6 h = 3237.0 and 181.8 pmol h/L plasma respectively) relative to GT control (AUC_0–6 h = 1304.1 and 61.0 pmol h/L plasma respectively). In vitro digestive recovery was higher for EGC and epicatechin (EC) (～51–53%) relative to EGCG and epicatechin gallate (ECG) (<20%) and was modestly enhanced in GT + S and GT + S + AA formulations. Accumulation of EGC, EGCG and ECG by Caco-2 cells was significantly (P < 0.05) higher from GT + S + AA compared to other formulations while retention of catechins was enhanced in presence of ascorbic acid. These data suggest that formulation with sucrose and ascorbic acid may improve catechin bioavailability by enhancing bioaccessibility and intestinal uptake from tea.     

Daily green tea extract supplementation reduces prothrombotic and inflammatory states in dialysis patients

Cardiovascular disease (CVD) remains the most common cause for excess morbidity and mortality in patients with chronic kidney disease (CKD) and end stage renal disease (ESRD) under chronic dialysis. ESRD patients have increased oxidative stress and endothelial dysfunction alongside increased levels of inflammation related proteins, which has prompted the exploration of anti-inflammatory and antioxidant treatments to improve outcomes. As green tea is increasingly well recognized for its antioxidant properties, we probed the effect of consumption of 1 capsule daily of green tea as a commercially available, decaffeinated green tea capsule (1 g, catechin content 68 mg) for 6 months on fibrinogen and inflammation in dialysis patients. Chronic hemodialysis patients (N = 25) were recruited and fibrinogen, FDP-D-dimer, high sensitivity (hs) CRP and the mononuclear cell protein expression of p22^phox, were assessed before, i.e. baseline and after 6 months of ingestion of 1 green tea capsule per day. After 6 months of daily green tea capsule ingestion, dialysis patients showed reduced protein expression of p22^phox (p < 0.0001), reduced hsCPR (p = 0.032) and fibrinogen (p = 0.022) levels and increased FDP-D-dimer (p = 0.0019) compared to their values at baseline. These results document lower oxidative stress and inflammation with green tea capsule ingestion and suggest a likely positive impact of green tea treatment on the atherosclerotic process of ESRD patients under dialysis.     

In vitro human skin permeation and cutaneous metabolism of catechins from green tea extract and green tea extract-loaded chitosan microparticles

Catechins are major antioxidants in green tea (Camellia sinensis or Camellia assamica), but because they do not permeate the skin well, the application of green tea in cosmetic products has so far been limited. This study aims to evaluate the cutaneous absorption of catechins from an extract of green tea and from a green tea extract-loaded chitosan microparticle. The catechin skin metabolism was also examined. The results suggest that chitosan microparticles significantly improve the ability of catechins to permeate skin. The cutaneous metabolism of the catechins significantly affected their permeation profiles. Epicatechin (EC) and epigallocatechin (EGC) penetrated the skin more than epigallocatechin gallate (EGCG) and epicatechin gallate (ECG). The galloyl groups in EGCG and ECG were enzymatically hydrolysed to EGC and EC, respectively. Dehydroxylation of catechins was also observed. Chitosan microparticles effectively prevented enzymatic changes of the catechins; therefore, chitosan microparticles are here found to be the promising carriers for enhancing the skin permeation.     

乌龙茶加工过程中儿茶素的稳定性及化学变化

研究半发酵乌龙茶加工对活性化学成分儿茶素的影响。采摘的鲜叶立即在严格控制的加工条件下进行加工,分析每道加工工艺操作所制茶叶的儿茶素总量和各种儿茶素单体(－)-儿茶素、(－)-表没食子儿茶素、(＋)-儿茶素、(－)-表儿茶素、(－)-表没食子儿茶素没食子酸酯、没食子儿茶素没食子酸酯、(－)-表儿茶素没食子酸酯、(－)-儿茶素没食子酸酯的含量。研究表明,在乌龙茶加工过程中,儿茶素总含量略有下降,约为10%;儿茶素单体变化不一。在加工工艺中,做青、杀青和干燥对儿茶素含量影响较大。为使乌龙茶中儿茶素的潜在健康效益最大化,这3 道工艺应在茶叶生产中加以考虑并做到最优化,以期为乌龙茶加工研究提供一定的参考依据。     

野化白云黄茶闷黄过程中的品质成分变化及其菌种分离鉴定

为了掌握野化白云茶树鲜叶加工成黄茶加工过程的影响因素,该研究对比分析了野化白云茶加工的黄茶、烘青绿茶、半烘半炒绿茶的主要品质成分;分析了野化白云茶鲜叶加工黄茶过程中的依工艺阶段取8个茶样（茶青、杀青叶、揉捻叶、再炒叶（除水叶）、闷黄24 h、闷黄48 h、闷黄72 h、毛茶）可培养菌种的变化。野化白云茶与烘青绿茶、半烘半炒绿茶比较,除氨基酸、咖啡碱含量略有下降外,水浸出物含量分别高出4.80%、7.11%,可溶性糖含量分别高出0.41%、0.36%,茶多酚含量、儿茶素总含量呈下降趋势,酯型儿茶素总量下降21.62 mg/g、19.08 mg/g,EGCG下降9.99 mg/g、6.98 mg/g,GCG下降14.23 mg/g、13.80 mg/g,以上成分变化均差异显著（p<0.05）;非酯型儿茶素组分中除C减小2.49 mg/g、1.75 mg/g（p<0.05）外,其余非酯型儿茶素组分和总含量变化差异不显（p>0.05）;闷黄24 h,酵母菌数量明显,至72 h霉菌成优势菌种（3.5×105 CFU/g）;分离鉴出8株微生物,包括1株拟盘多毛孢属,4株青霉属,1株枝孢属及2株酵母菌属,其中Penicillium exsudans菌首次从茶叶加工中和茶叶中分离并鉴定。说明闷黄使茶叶水溶性物质、可溶性糖增加,茶多酚、酯型儿茶素下降,闷黄阶段有微生物繁殖并参与黄茶品质的形成。