Epimerisation of tea polyphenols in tea drinks

The present study was carried out to quantify green tea epicatechin (GTE) derivatives and to investigate the origin of epicatechin epimers present in 18 selected canned or bottled tea drinks. The major GTEs present in tea are (?)‐epigallocatechin gallate (EGCG), (?)‐epigallocatechin (EGC), (?)‐epicatechin gallate (ECG) and (?)‐epicatechin (EC). HPLC analysis showed that the content of total GTEs was lower (16.4–268.3 mg l^?1) in the canned and bottled tea drinks than in tea traditionally prepared as a beverage in a cup or teapot (3–5 g l^?1). The major finding was that they contained higher levels of epicatechin epimers, namely (?)‐gallocatechin gallate (GCG), (?)‐gallocatechin (GC), catechin gallate (CG) and (?)‐catechin (C), than of GTEs, ranging from 7.6 to 331.8 mg l^?1. To investigate the origin of these epimers, GTEs were extracted from longjing green tea and autoclaved at various temperatures for 10–60 min. It was found that at least 50% of GTEs were epimerised to their corresponding epimers when autoclaved at 120 °C for 20 min. It is concluded that epicatechin epimers in tea drinks are not originally present in green tea leaf but are instead derived from thermal conversion of GTEs. Copyright © 2003 Society of Chemical Industry     

Flavanol Composition and Caffeine Content of Green Leaf as Quality Potential Indicators of Kenyan Black Teas

The flavanol composition and caffeine content of green tea leaf, black tea quality parameters of theaflavins, thearubigins, liquor brightness and total colour varied more among clones than with time of the year. In green leaf, either (-)epicatechin gallate or (-)epigallocatechin gallate was the dominant flavanol present. Regression analysis of tasters' preferences for black teas against green leaf chemical components showed positive and significant correlations for (-)epicatechin gallate (r = 0·498, P ⩽ 0·05 for taster A; r = 0·665, P ⩽ 0·01 for taster B, and r = 0·678, P ⩽ 0·01 for both tasters' overall ranking), (-)epigallocatechin gallate (r = 0·513, P ⩽ 0·05 for taster B; r = 0·532, P ⩽ 0·05 for both tasters' overall ranking and caffeine (r = 0·523, P ⩽ 0·05 for taster A; r = 0·657, P ⩽ 0·01 for taster B; and r = 0·686, P ⩽ 0·01 for both tasters' overall ranking). Similar regressions against black tea theaflavins, thearubigin content, liquor brightness and total colour were not significant. The results suggest that the green leaf chemical components, (-)epicatechin gallate, (-)epigallocatechin gallate and caffeine could be used as quality potential indicators during clonal selection and propagation. © 1997 SCI.     

GREEN TEA CATECHINS AS INHIBITORS OF OXIDATION OF MEAT LIPIDS

Antioxidant activity of green tea catechins, namely, epicatechin (EC), epigallocatechin (EGC), epicatechin gallate (ECG) and epigallocatechin gallate (EGCG) was evaluated in a meat model system. The inhibitory effect on 2-thiobarbituric acid (TBA) values of catechins was concentration dependent, being highest at 200 mg/kg. At 200 mg/kg, the antioxidant activity of catechins as evaluated by TBA values was EGCG ≈ ECG > EGC > EC. The hexanal and pentanal contents in the headspace volatiles of meats treated with catechins at 200 mg/kg was also evaluated. These results indicated that EGCG was most active followed by ECG and EGC; EC was least effective. In comparison with α-tocopherol, catechins were generally more effective in controlling oxidation of meat lipids. Based on TBA results, butylated hydroxytoluene (BHT) was less effective, but exerted a better effect than catechins in prevention of oxidation as evidenced by the hexanal/pentanal data.     

Polyphenolic composition and in vitro antioxidant activities of native‐ and tannase‐treated green tea extracts

The antioxidant activities of native‐ and tannase‐treated green tea extracts along with their major polyphenol components were investigated. The polyphenolic content and composition of the tea before and after tannase treatment were determined by liquid chromatography coupled with mass spectrometry (LC‐MS). Approximately 99% of the (?)‐epigallocatechin gallate (EGCG) and (?)‐epicatechin gallate (ECG) in green tea extract were converted by tannase to (?)‐epigallocatechin (EGC) and (?)‐epicatechin (EC), respectively, after 30 min. Biotransformed green tea exhibited a significantly higher DPPH˙ radical scavenging activities than native green tea (EC₅₀ value of 0.024 ± 0.001 and 0.044 ± 0.001 mg mL^?1, respectively). Kinetic parameters such as scavenging rate and stoichiometry were calculated. The rate of DPPH˙ radical scavenging activities for tannase‐treated green tea extract was shown to be higher than native green tea extract.     

Impact of heating on chemical compositions of green tea liquor

Heating pasteurization changed flavour of green tea liquor and it was a technical barrier in ready-to-drink tea production. The effect of heating on chemical compositions of green tea liquor was investigated by high performance liquid chromatography and gas chromatography–mass spectrometry. With increase of heating temperature from 85 °C to 120 °C, the green tea liquor became darker and less green, but deeper yellow in colour. During the heating, epigallocatechin gallate, epigallocatechin, epicatechin and epicatechin gallate partially epimerized, and concentration of total catechins decreased. Twenty volatiles were identified and concentrations of pentanol, cis-3-hexenol, linalool oxide I, linalool oxide II and β-ionone were decreased while phenylacetaldehyde, linalool, linalool oxide III, α-terpineol and indole increased. The decline of volatiles with pleasant odours and increase of some volatiles with unpleasant odours, such as indole (animal-like) and α-terpineol (faint ammoniacal), was considered to be responsible for the change in flavour of green tea liquor. Treatment at 85 °C caused fewer changes in liquor colour and concentrations of catechins and volatiles and it is recommended that extraction and pasteurization of canned ready-to-drink green tea should be carried out at 85 °C or less.     

陕西省不同产地绿茶中6种活性成分含量的比较

目的：采用高效液相色谱法HPLC,对陕西不同产区(安康、汉中和商洛)绿茶中茶氨酸(theanine)、没食子酸(gallic acid,GA)、咖啡碱(caffeine,CAF)、表没食子儿茶素(epicatechin gallate,EGC)、表儿茶素没食子酸(epicatechin gallate,ECG)及表没食子儿茶素没食子酸酯(epigallocatechin gallate,EGCG)的含量进行分析比较。方法：绿茶经热水提取后采用HPLC测定上述6种活性成分的含量,色谱柱(250mm×4.6mm,5μm),流动相为0.05%三氟乙酸溶液(A)、乙腈(B)梯度洗脱,流速为1mL/min,柱温25℃,检测波长：190nm和280nm。结果：3个产区中6种组分的含量在0.325～47.061mg/g之间,产区间各组分含量存在显著性差异(P＜0.05),其中汉中绿茶中GA的平均含量最高,安康绿茶中茶氨酸、CAF、ECG、EGC、EGCG的平均含量最高。     

Quantitative and Qualitative Portrait of Green Tea Catechins (Gtc) Through Hplc

Green tea (Camellia sinensis) is a prosperous source of polyphenols, especially catechins. In the current research, an effort was made to optimize the extraction conditions for maximum yield of catechins from the local green tea Qi-Men. For the purpose, three different solvents were used, i.e., aqueous ethanol (50%), aqueous methanol (50%), and water at different time intervals (20, 40, and 60 min). Green tea catechins were quantified through HPLC using a C18 column and UV detector. The antioxidant activity of green tea catechins was measured through in vitro tests including DPPH radical scavenging ability and antioxidant activity. Results showed that extraction through aqueous ethanol resulted in maximum yield of green tea catechins (17400 ± 0.19 mg/100 g green tea leaves. Moreover, epigallocatechin, epigallocatechin gallate, epicatechin gallate, and epicatechin ranged from 4.26 ± 0.09 to 6.4 ± 0.2, 12.1 ± 0.123 to 17.7 ± 0.3, 1.32 ± 0.03 to 1.81 ± 0.02, 5.48 ± 0.099 to 8.6 ± 0.2 g/100 g of dry-extract, respectively. Furthermore, highest antiradical (80.65 ± 3.69%) and antioxidant activity (67.12 ± 3.08%) were observed in catechins extracted through aqueous ethanol.     

“紫娟”茶中的EGCG3"Me成分研究

"紫娟"是云南大叶群体种中的一种稀有茶树品种,因富含花青素成分其新鲜嫩梢的芽、叶、茎均呈现紫色,从而具有很高的科研价值和经济开发潜力,当今其功能化学成分的研究已成为了茶学研究热点之一。本文采用HPLC以及LC-MS技术首次报道分析了茶树特异品种"紫娟"中的表没食子儿茶素-3-O-(3-O-甲基)没食子酸酯(EGCG3"Me)成分及高EGCG3"Me含量"紫娟"茶的加工工艺。结果表明,茶树特异品种"紫娟"鲜叶中含有EGCG3"Me成分(含量为1.05%);在一芽五叶新梢不同嫩度的鲜叶中,第三叶的EGCG3"Me含量最高(含量为1.24%);采用蒸青绿茶或者晒青绿茶的加工工艺可以使成茶有效保留鲜叶中的EGCG3"Me成分。此外,还分析比较了普通炒青绿茶与"紫娟"绿茶中主要儿茶素成分的含量差异,发现较高的ECG和EC含量而较低的EGCG含量是茶树特异品种"紫娟"绿茶中儿茶素组成的主要特点。     

Comparison of catechins and volatile compounds among different types of tea using high performance liquid chromatograph and gas chromatograph mass spectrometer

As tea is one of the most popular beverages consumed worldwide, it is important for customers and business investigators to develop an easy and reliable method to discriminate between different types of teas from each other. A total of eighty‐seven types of various white, green, oolong, black and Puer teas were collected from the major tea estates in China, and their catechin contents and volatile compounds were compared by high performance liquid chromatograph and gas chromatograph mass spectrometer. It was found green tea contained the highest concentrations of total catechins, (?)‐epicatechin gallate (ECG) and (?)‐epigallocatechin gallate (EGCG), while oolong teas contained the highest concentrations of (?)‐epigallocatechin (EGC) among these five types of teas. The aroma composition and their quantities in different types of teas varied quite widely. The concentration of ECG, EGCG, pentanal, hexanal, methyl jasmonate, indole, (E,E)‐2,4‐hexadienal and 1,2,3‐trimethoxybenzene was shown to be different and could be used to discriminate white, green, oolong, black and Puer teas. The result showed that different types of teas could be partially classified by cluster analysis using index of individual catechins and volatile components.     

A simple and rapid turbidimetric method for determining catechins in beverages

We have developed a simple and rapid turbidimetric method to determine catechins based on the fact that many polyphenols produce hydrogen peroxide in an alkaline environment and that hydrogen peroxide oxidises cerium to generate cerium oxide precipitates. Four catechins (epicatechin, epigallocatechin, epicatechin gallate, epigallocatechin gallate) aggregated with these precipitates to form massive precipitates with increased turbidity. The catechins solution (0.18 mL) was mixed with 0.02 mL of 1% CeCl₃ solution, and absorbance (650 nm) was measured immediately after agitation for 3 min using a spectrophotometer. Absorbance was strongly correlated (0.99) with the concentration of each catechin compound. For commercially bottled green tea, the estimated catechin content determined using this turbidimetric method showed better correlation with the content determined by high‐performance liquid chromatography than that determined using ferrous tartrate method, which is the official Japanese method for determining the tannin content of green tea.     

The development of a suitable manufacturing process for ‘Benifuuki’ green tea beverage with anti‐allergic effects

Epigallocatechin‐3‐O‐(3‐O‐methyl) gallate (EGCG3″Me) has been reported to inhibit type I allergy better than epigallocatechin gallate (EGCG), a major catechin in tea leaves (Camellia sinensis L). We examined the effects of extraction and sterilization on the catechin content and histamine release from mast cells, as a representative reaction of early phase allergy, in the manufacture of ‘Benifuuki’ green tea beverage. Among various varieties of tea, the cultivar ‘Benifuuki’ contains approximately 2% of EGCG3″Me. Ester‐type catechins and their epimers increased with the increased extraction temperature of the tea. A tea infusion, extracted at 90 °C, strongly inhibited histamine release from mast cells. Furthermore, sterilization affected the catechin content in the manufactured green tea beverage. Sterilization at high temperature promoted the isomerization of catechins and the sterilized green tea beverage had a strong inhibitory effect. When EGCG3″Me, EGCG, epicatechin‐3‐O‐gallate (ECG) and their epimers, GCG3″Me (gallocatechin‐3‐O‐(3‐O‐methyl) gallate), GCG (gallocatechin‐3‐O‐gallate) and CG (catechin‐3‐O‐gallate) were compared, the anti‐allergic effect of GCG3″Me was strongest, and the order of activity was GCG3″Me > EGCG3″Me > GCG > EGCG. We consequently suggest that it was necessary to extract components from tea at the highest temperature possible, and to pasteurize under retort conditions (118.1 °C, 20 min), to manufacture functional green tea beverage with an anti‐allergic action. Copyright © 2005 Society of Chemical Industry     

The effect of ascorbic acid,citric acid and low pH on the extraction of green tea: How to get most out of it

Green tea seems to have a positive impact on health due to the therein-found flavanols. The amounts of these substances depend on tea preparation. In this paper, the influence of steeping time (3–7 min) and temperature (70–100 °C) on the content of the main flavanols in green tea (epicatechin EC, epicatechin gallate ECg, epigallocatechin EGC, and epigallocatechin gallate EGCg) is presented. Furthermore, additives (phosphate buffers, ascorbic acid in different amounts, and citric acid) are used to investigate the influence of pH, antioxidative, and chelating agents, simulating the addition of lemon juice or pure vitamin C.     

ANTIOXIDANT PROPERTIES OF POLYPHENOLS EXTRACTED FROM GREEN AND BLACK TEAS

The catechins, including epicatechin gallate (ECG), epigallocatechin (EGC), and epigallocatechin gallate (EGCG), and the theaflavins, including theaflavin (TF), theaflavin monogallate (TF-1), and theaflavin digallate (TF-2), were extracted from green tea and black tea, respectively. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging ability, superoxide-scavenging efficiency, and lipid oxidation-inhibition ability of the pure compounds listed above as well as epicatechin (EC), carnosol, carnosic acid, and butylated hydroxytoluene (BHT) were investigated.
The DPPH radical-scavenging ability of the catechins was EGCG > ECG > EGC > EC and of the theaflavins was TF-2 > TF-1 > TF. EGCG, ECG, EGC, TF-2, TF-1, and TF showed higher DPPH radical- and superoxide-scavenging abilities than carnosol, carnosic acid, and BHT. EGCG, ECG, EGC, carnosol, and carnosic acid showed higher lipid oxidation-inhibition activity, as measured by the Rancimat method, than BHT and theaflavins.     

A novel technique for chitosan microparticle preparation using a water/silicone emulsion: green tea model

Many effective methods such as spray drying, coacervation, ionic gelation, solvent evaporation and sieving have been suggested for entrapping bioactive compounds into micro‐ or nanoparticles. However, those methods still have some limitations owing to high temperature requirement, difficulty in particle harvesting or low entrapment for uncharged molecules. In this study, a novel chitosan microparticle preparation method was developed using water‐in‐silicone emulsion technique with green tea extract as a model active compound. Chitosan microparticles of diameter <5 μm were obtained from 2% chitosan solution with tripolyphosphate (TPP) solution as the hardening agent. The size and properties of the particles appeared to depend on several parameters such as TPP, emulsifier concentrations and pH. High concentration of emulsifier led to low encapsulation and particle aggregation. Entrapment efficiency of chitosan microparticles was improved with lower pH of the tripolyphosphate solution [59.94 ± 3.97 of epigallocatechin gallate (EGCG)] while slowing release of catechins. Epigallocatechin and epicatechin were released almost completely within 2 h under acidic condition whereas EGCG and epicatechin gallate were slowly released. In neutral condition, release of catechins depended on their molecular stabilities. The stabilities of catechins loaded in chitosan microparticles were varied under various temperatures. The degradation of tea catechins increased with temperature. However, the degradation of tea catechins loaded in chitosan microparticles was less than that of free catechins. Thus, the new technique for preparing chitosan microparticles containing heat‐sensitive water soluble green tea extract was successfully developed. The technique is suitable for micro‐encapsulation of hydrophilic compounds into chitosan microparticles with the ease of harvesting technique.     

槠叶种制成的不同茶类品质特性和抗氧化功能

摘 要: 目的 探究祁门槠叶种制作的不同茶类的感官品质和理化特性。方法 按生产性工艺制作不同种类茶样,感官审评法评定各茶类的基本品质特征,用氨基酸分析仪、液相色谱和气质联用方法分析关键品质成分,用体外模拟方法测定不同茶类的自由基清除力,SPSS用于统计分析。 结果 卷形红茶芳樟醇及其氧化物,香叶醇含量仍然较高,并产生6-甲基-5-庚烯-2-酮, 2,2,6-三甲基-6-乙烯基四氢-2H-呋喃-3-醇等烘干型红茶中很少的甜香成分,红茶干燥中烘炒制结合应成为一种普遍推行的技术。安茶滋味醇浓,但没有一般黑茶那种木腐味和陈香。3种茶叶中绿茶表没食子儿茶素没食子酸酯（EGCG）含量最高,但黑茶中表没食子儿茶素（EGC）和没食子儿茶素没食子酸酯(GCG)含量高于红茶和绿茶。1,1-二苯基-2-三硝基苯肼（DPPH）自由基、超氧自由基和羟自由基试验表明,大多数情形下安茶（黑茶）自由基清除力最强,但3种茶叶差异不明显,总还原力试验显示绿茶>黑茶>红茶。醇提取物和水提取物对比,抗氧化力差异没有表现一致性的规律。     

SEPARATION OF INDIVIDUAL CATECHINS FROM GREEN TEA USING SILICA GEL COLUMN CHROMATOGRAPHY AND HPLC

Crude catechin mixtures from green tea were separated into six fractions using a silica gel column chromatography and a chloroform-methanol-water (65:35:10, v/v/v, lower phase) solvent system. Fraction I was free of catechins, fraction II contained epicatechin (EC), fraction III had epicatechin and epigallocatechin (EGC), fraction IV possessed EGC, fraction V contained EGC, epicatechin gallate (ECG) and epigallocatechin gallate (EGCG), and fraction VI had EGCG. EC and EGC were separated from fractions II, III and IV using HPLC with a RP-18 semipreparative column and a water-dimethylformamide-methanol-acetic acid (157:40:2:1, v/v/v/v) solvent system. For isolation of EGC, ECG and EGCG from fractions V and VI a water-acetonitrile-methanol-acetic acid (159:36:4:1, v/v/v/v) solvent system was employed. Chemical structures of purified catechins were further confirmed by ESI-MS.     

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.     

3 种名优绿茶的特征滋味成分研究及种类判别

采用高效液相色谱与高分辨质谱相结合的方法对西湖龙井、黄山毛峰和信阳毛尖3 种名优绿茶茶汤中的共有特征滋味成分进行研究,并据此进行种类判别分析。通过研究,确定了以表没食子儿茶素没食子酸酯、咖啡因、表儿茶素没食子酸酯、表没食子儿茶素等为主的15 种共有特征滋味成分,并建立了其相应的特征指纹图谱。利用15 种共有特征成分峰的相对峰面积,建立了多元化典型判别函数,采用逐步判别分析技术对31 个绿茶样品进行了很好的种类判别,判别正确率为100%。     

不同产地扁形绿茶的品质成分差异分析

为了系统和全面地了解扁形绿茶的品质特征,分析来自湖北大悟、浙江磐安、安徽歙县、贵州湄潭、四川峨眉山5个产地扁形绿茶样品的感官品质、色差以及茶多酚、游离氨基酸、水浸出物、咖啡碱、可溶性糖等主要化学成分含量差异。结果表明,龙井茶干茶L1值(亮度值)与b1值(黄蓝色度值)分别达到38.20和19.35,均显著(P<0.05)高于其他4个产地的扁形绿茶,大方茶茶汤L3值最高。大方茶和湄潭翠芽平均水浸出物总量超过48.00%,显著(P<0.05)高于孝感龙剑茶、龙井茶及竹叶青茶。湄潭翠芽的游离氨基酸、咖啡碱、儿茶素、表儿茶素没食子酸酯、儿茶素没食子酸酯含量均最高,表没食子儿茶素没食子酸酯含量以孝感龙剑茶最高。基于各品质成分的相对含量建立的偏最小二乘法判别分析模型成功区分了5个产地的扁形绿茶样品。同时,利用偏最小二乘法判别分析变量重要性因子值筛选到11种关键成分,分别是干茶E1(综合色差值)、干茶a1(红绿色度值)、干茶b1、表儿茶素没食子酸酯、没食子儿茶素没食子酸酯、咖啡碱、游离氨基酸、茶汤a3、茶汤b3、茶汤L3、茶汤E3,可作为区分5个产地扁形绿茶的品质指标。本研究结果将为扁形绿茶加工工艺和品质提升提供一定的科学参考。