Tea extraction methods in relation to control of epimerization of tea catechins

Effect of water temperature and ethanol concentration on epimerization and extractability of tea catechins was investigated. The results showed that epigallocatechin gallate (EGCG) and epicatechin gallate (ECG) were partially epimerized into gallocatechin gallate (GCG) and catechin gallate (CG), respectively, when tea catechins extract was heated in water solution at 100 °C for 2 h or dry tea was extracted in water at 100 °C. The epimerization of the catechins was inhibited if the tea catechins extract was heated as solid powder and the dry tea was extracted in 50% (v/v) ethanol or in water at 80 °C or below. When the dry tea was extracted in water, the extractability of catechins increased with the increase of extraction temperature up to 100 °C, but there was no statistically significant difference in total catechins between 80 °C and 100 °C. When teas were extracted using ethanol solutions, the highest extractability of total catechins was obtained in 50% (v/v) ethanol for dry tea and in 75% (v/v) ethanol for fresh tea leaf. In order to reveal the real profiles of tea catechins in teas to be tested, dry tea should be extracted in 50% (v/v) ethanol for 10 min, while fresh tea leaf should be extracted in 75% (v/v) ethanol for 10 min. For commercial extraction, temperature should be controlled at 80 °C if water is used as the solvent. Copyright © 2007 Society of Chemical Industry     

渥堆发酵过程中藏茶化学成分的变化

目的：阐明藏茶渥堆过程主要化学成分的变化规律。方法：以不同发酵阶段藏茶样为原料,研究发酵过程中藏茶水提物中可溶性固形物、可溶性糖、总多酚、总黄酮含量及水提物体外抗氧化活性的变化,并应用超高效液相色谱仪检测水提物中茶碱、咖啡碱、没食子酸和8种儿茶素类单体的含量,应用酶联免疫法测定水提物中茶黄素、茶红素和茶褐素的含量。结果：藏茶发酵过程中可溶性固形物含量先降低后略有回升,可溶性糖、总多酚、总黄酮含量逐渐降低,抗氧化活性逐渐减弱,咖啡碱呈波动状态且变化较小,没食子酸与儿茶素含量先上升再略有下降,没食子儿茶素没食子酸酯与表儿茶素含量先升高后降低,没食子儿茶素、表没食子儿茶素、表儿茶素没食子酸酯和表没食子儿茶素没食子酸酯含量均呈持续下降的状态,其中表没食子儿茶素没食子酸酯在黑毛茶中含量为39.69 mg/g,在成品茶中降至1.36 mg/g;茶红素含量先增加后降低,茶黄素含量变化很小,茶褐素含量逐渐升高（黑毛茶中3.12 g/100 g,成品茶中7.46 g/100 g）。结论：藏茶渥堆发酵过程中,可溶性糖、总多酚、总黄酮等含量降低,抗氧化活性减弱,茶碱、咖啡碱、儿茶素类单体等活性成分含量的变化趋势不一致,茶褐色含量逐渐升高。     

Antibacterial action on pathogenic bacterial spore by green tea catechins

Antibacterial effects of catechins, the major green tea polyphenols, were studied using Clostridium and Bacillus spores. Incubation with crude catechins decreased the number of C botulinum and C butyricum spores but not B cereus spores. Furthermore, the effects of six catechin derivatives on spores were investigated. (−)‐Epicatechin gallate (ECg), (−)‐epigallocatechin (EGC), (−)‐epigallocatechin gallate (EGCg) and (+)‐gallocatechin gallate (GCg) were more effective in decreasing C botulinum and C butyricum spore numbers than (+)‐catechin (C) and (−)‐epicatechin (EC). The vegetative growth of C botulinum and B cereus was inhibited by crude extracts of the catechins. Specifically, purified GCg and EGCg inhibited the vegetative growth of C botulinum and B cereus. The inhibitory effect of ECg on B cereus was similar to that of GCg. However, toxin‐production by B cereus was not inhibited by catechin. Damage to the membrane of C butyricum spores by catechin derivatives was shown using fluorescent microscopy. This study shows that low concentrations of catechins, although requiring a long exposure time, inhibited the growth of bacterial spores. However, the effects of the purified derivatives of the catechins were not the same and GCg and EGCg were found to be the most potent. Spores that are generally resistant to many disinfectants were sensitive to catechins. Copyright © 2005 Society of Chemical Industry     

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.     

不同类型沱茶品质与特征性成分分析

目的 探明不同类型沱茶的品质和特征性成分。方法 本文采用高效液相色谱、感官审评、Lab颜色模型等方法,比较分析供试沱茶的主要品质成分、干茶色泽和感官品质,结合多元统计学方法明确不同沱茶的特征化学物质组成。结果 云岭沱茶的感官审评综合得分高于其他沱茶,其汤色亮度和叶底嫩度优于其他沱茶,下关沱茶以儿茶素、表儿茶素、表儿茶素没食子酸酯含量较高,而茶黄素含量较低为特征,山城沱茶以茶红素含量较高,而水浸出物、咖啡碱、没食子酸含量较低为特征,云岭沱茶以茶黄素、茶多酚、表没食子儿茶素没食子酸酯、表没食子儿茶素、没食子酸、咖啡碱含量较高,而茶红素、茶褐素、儿茶素含量较低为特征,与沱茶感官审评得分呈显著正相关的生化成分有茶多酚、咖啡碱、没食子酸、茶黄素、表没食子儿茶素、表没食子儿茶素没食子酸酯。结论 不同类型沱茶生化成分的含量与组成特征差异较大。     

Potent antioxidative activity of non‐polyphenolic fraction of green tea (Camellia sinensis) – association with pheophytins a and b

The antioxidative activity of green tea‐derived polyphenols known as catechins has been extensively studied. However, the antioxidative activity of the non‐polyphenolic fraction of green tea has been poorly elucidated and is the subject of the present study. The non‐polyphenolic fraction of green tea showed a significant dose‐dependent suppressive effect against the autooxidation of linoleic acid. The chlorophyll‐related compounds pheophytins a and b showed similar antioxidative activities in the same assay, much higher than those of α‐tocopherol and the green tea catechin (−)‐epigallocatechin‐3‐gallate (EGCG). The non‐polyphenolic fraction of green tea and pheophytins a and b exhibited suppressive activities against superoxide anion (${\bf O}_{2}ˆ{‐}$ ) generation in mouse macrophages induced by 12‐O‐tetradecanoylphorbol‐13‐acetate, showing higher activities than that of EGCG. These results suggest that the non‐polyphenolic fraction of green tea leaves has potent antioxidative activity and that this activity is associated with pheophytins a and b. © 2000 Society of Chemical Industry     

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.     

Taste characterisation of green tea catechins

There has been interest in biological activities of green tea catechins. However, little is known about the taste characteristics of catechins. To assess the taste characteristics of catechins ((?)–epicatechin (EC), (?)–epicatechin gallate (ECg), (?)–epigallocatechin (EGC) and (?)–epigallocatechin gallate (EGCg)), sensory evaluations were performed. The taste intensity increased with increased catechins concentration. Among them, ECg showed the strongest taste. Catechins had tastes that consisted primarily of astringency and bitterness. Therefore, taste palatability decreased with increasing catechin concentrations. In line with taste intensity, taste palatability of ECg was the lowest. Taste sensor analysis and mouse behavioural assays showed same results. EC and ECg were more stable in Ringer’s solution than EGC and EGCg. Furthermore, taste cell responses to ECg that had the strong taste and better stability among catechins used were recorded. Some taste cells responded to ECg. This result suggests that ECg might be recognised by taste cells.     

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     

利用活性真菌转化简单儿茶素为酯型儿茶素

研究从茶树的鲜叶、茎部和茶树土壤中分离出活性真菌,通过利用天然微生物的转化能力,使简单儿茶素转化为功能性较强的复杂型酯型儿茶素。实验结果表明:筛选出了具有转化能力的活性菌种,能有效地将简单儿茶素转化为酯型儿茶素,而且其转化能力良好。并对此种活性真菌进行形态学和生理生化的初步检测,判断其属于丝状真菌藻状菌纲,毛霉目,内囊霉科,内囊霉属。     

不同叶位芽叶在工夫红茶加工过程中理化品质的变化

本实验以福鼎大白茶茶树品种鲜叶为原料,通过叶片微观结构观察和理化指标测定,探究工夫红茶加工过程中不同叶位芽叶的理化品质变化。结果发现：不同叶位芽叶的色泽从揉捻开始由绿逐渐变红,但第二叶的红变程度弱于其他部分;叶片结构和酚类化合物分析结果表明,第一、二叶的叶肉细胞及叶绿体结构均在揉捻时被破坏,说明第一、二叶均从揉捻阶段开始发酵;在加工过程中茶多酚质量分数、儿茶素类含量均下降,茶黄素含量和茶红素含量先增后减,之后趋于稳定,茶褐素含量一直增加,且均在揉捻时含量变化最大;第一叶与第二叶的儿茶素类总量、表没食子儿茶素、表没食子儿茶素没食子酸酯、表儿茶素没食子酸酯的氧化率及茶黄素含量差异不显著（P>0.05）,说明不同叶位叶片的发酵程度无明显差异;叶绿素a、叶绿素b含量以及叶绿素类总量均下降,脱镁叶绿素、脱镁脱叶绿醇基叶绿素含量均上升;加工过程中第二叶的叶绿素组分含量均显著高于第一叶（P<0.05）。本研究结果可为加工优质工夫红茶、提高茶树鲜叶的利用率提供一定的理论依据。     

模拟体外消化评定超微绿茶粉的冲泡饮用效果

测定了超微绿茶粉模拟体外消化前后多种物质的释放情况。结果表明,超微绿茶粉在体外消化过程中,游离氨基酸、总酚、总黄酮及大部分儿茶素组分释放量有所减少;咖啡碱释放量保持不变;可溶性蛋白、没食子酸、儿茶素没食子酸酯、没食子儿茶素、没食子儿茶素没食子酸酯、异槲皮素的释放量有所增加,不可萃取多酚有所释放。说明冲泡饮用超微绿茶粉有利于提高人体对茶叶的综合利用率。     

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     

青砖茶加工过程品质成分变化研究

目的研究青砖茶加工过程中各化学品质成分含量的变化,为青砖茶风味品质形成提供理论依据。方法以青砖茶发酵前、发酵后及成品为材料,比较分析样品水分、水浸出物、游离氨基酸、咖啡碱、茶多酚、儿茶素组分、茶黄素组分、粗纤维、黄酮、可溶性糖、可溶性果胶等含量变化。结果青砖茶加工过程中,水分含量基本不变,控制在9%左右;纤维素含量呈现上升趋势,成品达到(21.29±0.78)%;水浸出物、可溶性总糖、儿茶素、茶黄素-3'-没食子酸酯、茶黄素总量呈现下降趋势;黄酮含量总体较为稳定,为1.40%~1.83%;茶黄素-3,3'-双没食子酸酯基本稳定在1 mg/g左右;咖啡碱含量发酵后显著低于发酵前;氨基酸、茶多酚、表儿茶素、表儿茶素没食子酸酯、表没食子儿茶素、表没食子儿茶素没食子酸酯、儿茶素总量、茶黄素、茶黄素-3'-没食子酸酯、水溶性果胶均呈现下降后上升,但成品含量低于发酵前原料含量的趋势。结论青砖茶加工过程中不同生化成分变化趋势存在差异,成品与原料存在较大差异,具有粗涩味作用的成分含量降低,各成分间含量更加协调,为青砖茶醇和风味品质形成奠定基础。     

A reliable technique to identify superior quality clones from tea germplasm

Variations in the substrate level and enzyme activity of prominent south Indian tea germplasm were studied. The content of polyphenols, catechins (substrates) and polyphenol oxidase (PPO) showed variation, which influenced the final black tea quality. The enzyme PPO occurs in tea shoots and catalyzes the reactions between catechins to form theaflavins in the presence of oxygen. The catechins mainly epicatechin (EC), epigallo catechin gallate (EGCG), epicatechin gallate (ECG) and epigallo catechin (EGC) get mixed with PPO during the oxidation process to form quality constituents like theaflavins (TF), thearubigins (TR) and high polymerized substances (HPS). Theaflavins and their fractions such as simple theaflavin, theaflavin monogallate, (TFMG), theaflavin digallate (TFDG) in black tea are the essential quality constituents that are responsible for the liquor characteristics where as TR and high polymerized substances impart colour to the liquor. As oxidation of macerated leaves proceed through different stages of tea manufacture, a decline in PPO activity, polyphenol and catechin contents were observed. Data revealed that the oxidation reaction was faster during the initial stages of oxidation. During the period, oxygen consumption was higher and declined thereafter. Ratio between the enzyme (PPO) and its substrate (catechins) were used to characterize the quality potential of tea clones. An attempt has also been made to categorize prominent tea clones as high, moderate and average quality clones based on their enzyme substrate ratio. Theaflavin content (oxidation product) of different tea clones suggests that the ratio between PPO and catechins forms an important criterion which determines black tea quality. Results obtained were compared with standard clones of known high quality (CR-6017) and moderate quality (SA-6). The study reveals that the enzyme substrate ratio can be used to identify superior quality clones from the existing tea germplasm.     

茶叶活性成分的抗炎作用及其机制研究进展

茶叶中主要活性成分如茶多酚、茶氨酸、茶黄素具有较好的抗炎作用,且茶多酚效果最佳。虽然茶多酚的抗氧化作用被世界公认,但这并不是其发挥抗炎作用的唯一机制,其机制应与抑制炎症因子、调节相关炎症信号通路、抗氧化等综合作用有关。本文综述了茶叶及茶叶成分茶多酚、茶氨酸、茶黄素的抗炎作用及其机制,其中重点阐述了主要儿茶素组分表没食子儿茶素没食子酸酯(epigallocatechin gallate,EGCG)的抗炎机制,为茶叶抗炎作用的深入研究提供参考,为健康饮茶和茶叶抗炎功能产品的研发提供理论依据。     

基于UPLC-MS/MS同时分析福建乌龙茶和白茶中的儿茶素类物质

目的:建立超高效液相色谱-串联质谱法(UPLC-MS/MS)快速分析茶叶中儿茶素类物质如儿茶素(Catechin,C)、表儿茶素(Epicatechin,EC)、没食子儿茶素(Gallocatechin,GC)、表没食子儿茶素(Epigallocatechin,EGC)、表没食子儿茶素没食子酸酯(Epigallocatechin gallate,EGCG)等,对几种福建乌龙茶和白茶中的儿茶素类物质进行分析。方法:实验使用Waters T3 C₁₈色谱柱(250 mm×4.6 mm,5 μm),以0.1%(V/V)甲酸水溶液和甲醇为流动相对目标物进行梯度洗脱后,电喷雾正离子扫描方式下多反应监测(MRM)模式分析。结果:8 min内即可完成茶叶中8种儿茶素类物质的上机检测;儿茶素类物质的检出限为0.2~1.1 μg/L,线性方程的相关系数(r)为0.9966~0.9994。用建立的方法分析所采集的几种福建茶叶样品,表没食子儿茶素没食子酸酯(EGCG)相对于其它儿茶素类物质在茶叶中的含量最高,约占茶叶中儿茶素总量的60%。乌龙茶中儿茶素总量的含量范围为43.59~45.21 mg/g,白茶儿茶素总量的含量范围为50.52~70.71 mg/g。结论:建立的方法灵敏、准确、快速,研究结果可为福建茶叶产品的开发提供一定的科学依据。     

Polyphenols Extracted from Black Tea (Camellia sinensis) Residue by Hot‐Compressed Water and Their Inhibitory Effect on Pancreatic Lipase in vitro

Abstract: Polyphenols, retained in black tea wastes following the commercial production of tea beverages, represent an underutilized resource. The purpose of this study was to investigate the potential use of hot‐compressed water (HCW) for the extraction of pancreatic lipase‐inhibiting polyphenols from black tea residues. Black tea residues were treated with HCW at 10 °C intervals, from 100 to 200 °C. The resulting extracts were analyzed using high‐performance liquid chromatography‐mass spectrometry and assayed to determine their inhibitory effect on pancreatic lipase activity in vitro. Four theaflavins (TF), 5 catechins, 2 quercetin glycosides, quinic acid, gallic acid, and caffeine were identified. The total polyphenol content of extracts increased with increasing temperature but lipase inhibitors (TF, theaflavin 3‐O‐gallate, theaflavin 3′‐O‐gallate, theaflavin 3,3′‐O‐gallate, epigallocatechin gallate, and epicatechin gallate) decreased over 150 °C. All extracts inhibited pancreatic lipase but extracts obtained at 100 to 140 °C showed the greatest lipase inhibition (IC₅₀s of 0.9 to 1.3 μg/mL), consistent with the optimal extraction of TFs and catechins except catechin by HCW between 130 and 150 °C. HCW can be used to extract pancreatic lipase‐inhibiting polyphenols from black tea waste. These extracts have potential uses, as dietary supplements and medications, for the prevention and treatment of obesity. Practical Application: Active forms of lipase inhibitors can be recovered from black tea residues. They could be used as dietary supplements or medications.     

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.     

Influence of fermentation time on the development of compounds responsible for quality in black tea

The compounds responsible for tea quality, such as theaflavins (TFs) and thearubigins were found to increase with fermentation time. However, TF reached a maximum, declining subsequently. The water extract of black tea decreased with fermentation time. Caffeine concentration remained unchanged. The digallate equivalent of theaflavin, colour index and briskness index were found to peak at the optimum fermentation time. Polyphenols declined more quickly during the initial stages, followed by a steadily declining trend. Fermentation time had little impact on the gallic acid concentration. Among the catechins, epigallocatechin oxidized fastest, followed by epigallocatechin gallate and epicatechin gallate.