Extraction of caffeine and catechins using microwave-assisted and ultrasonic extraction from green tea leaves: an optimization study by the IV-optimal design

In this research, optimal conditions for extraction of caffeine and polyphenols were established from Iranian green tea leaves. In the first step, caffeine was extracted with efficacy about 86% versed to 4.5% of EGC + EGCG. The EGCG + EGC was extracted from partially decaffeinated green tea leaves through microwave-assisted extraction (MAE) and ultrasound-assisted extraction (USE) with efficiency levels of 95 and 85%, respectively. The best results for the MAE process were obtained with 7.8 min and three number of extraction cycles and for the USE process were as followed: time 57 min, temperature 65 °C, and the number of extraction cycles 3. The total phenol content values at the best conditions of MAE and the USE processes were 125 ± 5 and 96 ± 6 mg gallic acid/g DW. The 50% inhibition (IC₅₀) on 1,1-diphenyl-2-picrylhydrazyl (DPPH) were 56 and 66 mg/g of phenol for the MAE and USE processes.     

Factors affecting the levels of catechins and caffeine in tea beverage: estimated daily intakes and antioxidant activity

Increased public awareness of health benefits of green tea is generally based on the high polyphenol content of tea leaves and the resulting beverage. A number of factors, such as species, season, agronomic condition and age of the leaves, are known to affect the composition of commercial teas. In the present study the effects of factors associated with domestic preparation and analytical methods, such as brewing time, concentration, solvent and type of tea product, on levels of catechins and caffeine, antioxidant activity and estimated daily intakes were investigated. There were large variations in the levels of total catechins: 43 and 117 mg g^?1 dry matter (DM) (brewed for 30 s and 5 min respectively); 72 and 161 mg g^?1 DM (extracted in boiling water and 50% acetonitrile respectively); 72 and 117 mg g^?1 DM (a tea bag and tea leaves respectively). The effects on caffeine content were comparatively smaller. These variations consequently led to considerable variations in estimated daily intakes based on three cups (600 ml), ranging between 538 and 2014 mg g^?1 DM of total catechins and between 103 and 466 mg g^?1 DM of caffeine. The antioxidant activity was highest (26 680 µmol g^?1 DM) for tea leaves brewed for 5 min and lowest (10 110 µmol g^?1 DM) for a tea bag product brewed for 1 min. Copyright © 2005 Society of Chemical Industry     

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     

Fenvalerate residue level and dissipation in tea and in its infusion

Fenvalerate is a non-systemic insecticide/acaricide used in controlling a wide range of pests, including those resistant to organochlorine, organophosphorus and carbamate insecticides. The study investigated the dissipation behaviour (residue level) of fenvalerate in tea and its transfer during infusion. Fenvalerate was applied on tea crop at two dosages, 100 and 200?g a.i.?ha^?1 (recommended and double the recommended) in the dry and wet seasons under field conditions. Samples (green tea shoots, made tea, its infusion and spent leaves) were analysed for fenvalerate by high-performance liquid chromatography using diode array detection. The residue dissipated faster in the wet season than in the dry season. Seven days after the treatment (normal round of plucking) the residues observed in the green shoots at the two dosages were 0.5?±?0.01, 1.1?±?0.01 and 0.4?±?0.02, 0.9?±?0.01?mg?kg^?1 in the dry and wet seasons, respectively. During processing of green tea shoots to made tea a 30–40% loss of residue was observed. The transfer of residue from made tea to infusion was in the range 10–30% for both seasons, whereas 50–70% of the residues remained in the spent leaves. However, the degradation rate in both seasons followed first-order kinetics. The half-lives were in the range of 2–3 days for green shoots and made tea in both seasons.     

Artificial neural network modeling and optimization of ultrahigh pressure extraction of green tea polyphenols

In this study, the ultrahigh pressure extraction of green tea polyphenols was modeled and optimized by a three-layer artificial neural network. A feed-forward neural network trained with an error back-propagation algorithm was used to evaluate the effects of pressure, liquid/solid ratio and ethanol concentration on the total phenolic content of green tea extracts. The neural network coupled with genetic algorithms was also used to optimize the conditions needed to obtain the highest yield of tea polyphenols. The obtained optimal architecture of artificial neural network model involved a feed-forward neural network with three input neurons, one hidden layer with eight neurons and one output layer including single neuron. The trained network gave the minimum value in the MSE of 0.03 and the maximum value in the R² of 0.9571, which implied a good agreement between the predicted value and the actual value, and confirmed a good generalization of the network. Based on the combination of neural network and genetic algorithms, the optimum extraction conditions for the highest yield of green tea polyphenols were determined as follows: 498.8 MPa for pressure, 20.8 mL/g for liquid/solid ratio and 53.6% for ethanol concentration. The total phenolic content of the actual measurement under the optimum predicated extraction conditions was 582.4 ± 0.63 mg/g DW, which was well matched with the predicted value (597.2 mg/g DW). This suggests that the artificial neural network model described in this work is an efficient quantitative tool to predict the extraction efficiency of green tea polyphenols.     

Vesicle-enhanced liquid-phase pulsed discharge extraction of polyphenols from green tea leaves

A vesicle-enhanced liquid-phase pulsed discharge (V-LPD) was proposed to extract polyphenols from green tea leaves. The optimal extraction conditions obtained by the process optimization were 3% (w/v) dodecyl trimethy lammonium bromide (DTAB)/sodium dodecyl sulfate (SDS) vesicle, 10% (v/v) ethanol/vesicle, 66 mL/g liquid-to-solid ratio, 5 kV discharge voltage, and 2.5 min extraction time. Under these optimal conditions, the polyphenols yield was 124.650 ± 1.197 mg/g, which was observably higher than that by heat reflux extraction (HRE) (108.156 ± 1.437 mg/g) for 45 min and that by ethanol-enhanced LPD (116.123 ± 1.164 mg/g) for 4 min. The energy consumptions of V-LDP and ethanol-enhanced LPD were far lower than that by HRE. Similar chemical constituents were observed by HPLC analysis in three extraction methods, and the extracts obtained by V-LPD had the highest antioxidant capacity. Therefore, the V-LPD is a time-saving and energy-saving method for the extraction of polyphenols.Industrial relevanceGreen tea is considered to be an important source of polyphenols. The potential of V-LPD has been demonstrated as a time-saving and energy-saving alternative using vesicle solvents for the simultaneous cell disruption and strengthening diffusion of polyphenols from green tea leaves. The polyphenols obtained with V-LPD process can be used in an array of pharmaceutical, cosmetic and food applications.     

挤压膨化工艺参数对夏秋绿茶多酚含量及膨化度的影响

利用双螺杆挤压膨化机对夏秋绿茶进行挤压膨化,研究物料含水量、套筒温度以及螺杆转速对膨化绿茶粉中茶多酚含量以及膨化度的影响,并分析夏秋绿茶膨化前后浸出功能成分及微观结构的变化。结果表明:随物料含水量增加,茶多酚含量先减少后增加;增加套筒温度会增大茶多酚的浸出,当温度过高时会导致茶多酚的损失;随螺杆转速增大,茶粉破损程度加大,促进茶多酚的浸出。套筒温度、螺杆转速对挤出物膨化度的影响较小,物料含水量的影响相对较大。与未加工夏秋绿茶粉相比,加工后的夏秋绿茶粉中茶多酚、粗纤维、可溶性总糖含量降低,茶氨酸含量增高。夏秋绿茶膨化后,其表面结构变得平整光滑,且机械力作用使物料自身化学键断裂。     

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.     

Investigation of the presence and significance of theanine in the tea plant

Theanine was determined in black teas and tea plants by a combination method of thin-layer chromatography (t.l.c.) and densitometry. It was shown that in 20 samples of black tea from different qualities and sources the amount varied from 0.33 to 1.59 g 100 g^?1 dry wt. The highest quality black teas possessed the lowest amount of theanine, but it is subject to chemical degradation during black tea manufacture. The investigation of physiological function of theanine in the tea plant showed that during germination the theanine level reached a maximum after 45 days. This indicates that at this period of the growth of the tea plant theanine acted as a source of nitrogen and as a starting point for the synthesis of the carbon skeletal compounds of the tea plant. Theanine was found to exist in all parts of the tea plant but it accumulated more in young and active tissues and also in younger plants, which emphasises its metabolic role in the tea plant. The distribution of theanine in the shoots indicated that the first leaf was the principal site for the synthesis of polyphenolic compounds from theanine.     

H NMR-based metabolomic characterization during green tea (Camellia sinensis) fermentation

The metabolic behavior of green tea (Camellia sinensis) during tea fermentation was characterized by ¹H NMR spectroscopy coupled with multivariate statistical analysis to provide comprehensive information on changes in metabolites induced by tea fermentation. Fourteen tea metabolites of epicatechin (EC), epigallocatechin (EGC), epicatechin-3-gallate (ECG), epigallocatechin-3-gallate (EGCG), theanine, alanine, acetate, quinate, glutamate, caffeine, sucrose, glucose, and gallate, as identified by ¹H NMR spectroscopy, were responsible for metabolic differentiation between green tea and fermented tea by principal component analysis. During tea fermentation, levels of EC, EGC, ECG, EGCG, quinate, caffeine, and sucrose were decreased, whereas gallate and glucose levels were increased. In particular, unique changes in caffeine and gallate levels were observed during tea fermentation, which caffeine and gallate levels have been shown to vary after tea fermentation among many reports to date. This study highlights that metabolomics with global profiling and a highly reliable and reproducible ¹H NMR spectroscopic data set can provide a better understanding of unique changes in tea metabolites during tea fermentation.     

Micronutrients (B,Co, Cu,Fe, Mn,Mo, and Zn) content in made tea (Camellia sinensis L.) and tea infusion with health prospect: A critical review

Tea (Camellia sinensis L.) is a perennial acidophilic crop, and known to be a nonalcoholic stimulating beverage that is most widely consumed after water. The aim of this review paper is to provide a detailed documentation of selected micronutrient contents, viz. boron (B), cobalt (Co), copper (Cu), iron (Fe), manganese (Mn), molybdenum (Mo), and zinc (Zn) in made tea and tea infusion. Available data from the literature were used to calculate human health aspect associated with the consumption of tea infusion. A wide range of micronutrients reported in both made tea and tea infusion could be the major sources of micronutrients for human. The content of B, Co, Cu, Fe, Mn, Mo, and Zn in made tea are ranged from 3.04 to 58.44 μg g^?1, below detectable limit (BDL) to 122.4 μg g^?1, BDL to 602 μg g^?1, 0.275 to 13,040 μg g^?1, 0.004 to 15,866 μg g^?1, 0.04 to 570.80 μg g^?1 and 0.01 to 1120 μg g^?1, respectively. Only 3.2 μg L^?1 to 7.25 mg L^?1, 0.01 μg L^?1 to 7 mg L^?1, 3.80 μg L^?1 to 6.13 mg L^?1, 135.59 μg L^?1 ?11.05 mg L^?1, 0.05 μg L^?1 to 1980.34 mg L^?1, 0.012 to 3.78 μg L^?1, and 1.12 μg L^?1 to 2.32 μg L^?1 of B, Co, Cu, Fe, Mn, Mo, and Zn, respectively, are found in tea infusion which are lower than the prescribed limit of micronutrients in drinking water by World Health Organization. Furthermore, micronutrient contents in tea infusion depend on infusion procedure as well as on the instrument used for analysis. The proportion of micronutrients found in different tea types are 1.0–88.9% for B, 10–60% for Co, 2.0–97.8% for Cu, 67.8–89.9% for Fe, 71.0–87.4% for Mn, 13.3–34% for Mo, and 34.9–83% for Zn. From the results, it can also be concluded that consumption of three cups of tea infusion per day does not have any adverse effect on human health with respect to the referred micronutrients rather got beneficial effects to human.     

Green tea from purple leaf coloured tea clones in Kenya- their quality characteristics

The Kenyan tea industry wishes to diversify its tea products, and in line with this, anthocyanin – rich teas were developed at the Tea Research Foundation of Kenya. These teas have purple-coloured leaves and the green colour is masked. In total, 12 accessions of the purple leaf coloured teas and 2 standard tea varieties were studied. Clones Hanlu and Yabukita are Chinese and Japanese tea varieties, respectively, known for good green tea, and they were used as reference standards. Little if any research had been done to characterize the quality of these purple leaf coloured teas and this study investigated their total polyphenols (TPP), catechins, caffeine, gallic acid and theanine. These are the major green tea quality parameters. Results showed that the new Kenyan tea clones had higher total polyphenols than had the reference standard tea varieties, which had 17.2% and 19.7% while the lowest among the Kenyan clones was 20.8%. On catechin quality index, K-purple and TRFK 91/1 showed high index values of 15.9 and 13.3, respectively, while clones TRFK 83/1 and 73/5 showed low index values of 0.74 and 1.0, respectively. Hanlu had the highest caffeine level with 2.42% while clones TRFK KS 3, TRFK KS 2 and TRFK 83/1 had relatively high caffeine levels among the purple leaf coloured teas, with 2.33%, 2.22% and 2.21%, respectively. Clone TRFK 73/5 had the lowest caffeine content, with 1.16%. Theanine analysis showed that most purple leaf coloured teas had more theanine than had the reference standard clones, except TRFK 83/1 and K-purple, which were lower than the reference standard clones. The implication of the green tea chemical quality parameters is also discussed. It is concluded that all the studied clones/varieties have above the minimum 14% of total polyphenols. Clones K-purple and TRFK 91/1 showed high green tea quality indices with the latter doubling with high levels of theanine; hence its highly recommended for green tea manufacture.     

Ethylamine content of fresh tea shoots and made tea determined by high performance liquid chromatography

The ethylamine contents of tea shoots and made teas were derived with o-phthalaldehyde and 2-mercaptoethanol, and quantified by high performance liquid chromatography. The coefficient of variation of this method was 1.97% from eight analyses. The levels of ethylamine in tea shoots declined markedly during growth and senescence after plucking; they also declined in the extracts of fresh tea during the incubation at 37°C, but this decrease was completely inhibited by amine oxidase inhibitors. The content of ethylamine in high grade green teas was higher than in low grade teas, but was almost identical in high and low grade black tea. The mean contents of ethylamine in 22 green teas and 17 black teas were 6.76±3.07 μmol g^?1 and 6.40±1.82 μmol g^?1, respectively.     

Detection of aflatoxins in tea samples based on a class‐specific monoclonal antibody

An enzyme‐linked immunosorbent assay (ELISA) using a monoclonal antibody was established to detect aflatoxin B₁ (AFB₁) in tea. The antibody was prepared from a hybridoma derived by fusing Sp2/0‐Ag14 myeloma cells and immunised spleen B cells. The effects from pH, ionic strength, and organic solvents on immunoassay were optimised and the 50% inhibition (IC₅₀) value was 0.057 ± 0.007 ng mL^?1. Spiked black and green tea samples at 10, 20 and 50 ng g^?1 levels of AFB₁ were detected with this proposed ELISA. The recoveries for black tea samples ranged from 68.5% to 117.7% and 73.5 to 114.3% for green tea samples. This immunoassay showed no cross‐reactions with other mycotoxin family but good recognition with related aflatoxins. These results indicate that the ELISA assay could be used as a screening method for aflatoxin detection in tea samples.     

A study on chemical estimation of pu‐erh tea quality

Chemical compositions and infusion colour differences of seven pu‐erh tea samples and their correlation to sensory quality were investigated. The results showed that the pu‐erh tea contained 37.1 mg g^?1 caffeine, 15.7 mg g^?1 amino acids, 67.0 mg g^?1 polyphenols and 8.41 mg g^?1 total catechins, on average. Among the 17 tested volatile compounds, n‐valeraldehyde was not detected. The most abundant volatile was β‐ionone and the next was linalool oxide II. Infusion colour analysis showed that the pu‐erh tea had deep hue with ΔE ranging from 66.8 to 79.2. Spearman's linear correlation analysis showed that total quality score (TQS) of the pu‐erh tea was significantly correlated to concentration of amino acids, linalool oxide II and infusion colour indicator ΔE. Five components were extracted from the 34 tested indicators by principal component analysis and were regressed on the TQS to produce six Pearson's linear regression equations for estimating sensory quality of pu‐erh tea, among which two were statistically significant, ie TQS = 57.47 ? 0.18geraniol + 0.33polyphenols ? 1.14n‐caproaldehyde ? 1.38linalool oxide I + 0.21caffeine (p < 0.01) and TQS = 57.42‐0.03Citral + 0.33polyphenols ? 1.14n ? caproaldehyde ? 1.40linalool oxide I + 0.20caffeine (p < 0.01). Copyright © 2004 Society of Chemical Industry     

从残次茶中制备茶多酚的关键技术研究

现今对残次茶的利用和茶多酚的提取工艺不完善,以至于造成残次茶的浪费。因此,研究从残次茶中提取茶多酚的关键技术是很有必要也很有意义的。试验以黑毛茶为原料,乙醇为提取溶剂,采用超声辅助提取的方法提取茶多酚,并通过响应面分析得到最佳工艺条件：超声功率70 W,乙醇体积分数70%,浸提温度65 ℃,浸提时间30 min,料液比1∶25（g∶mL）。在此条件下,茶多酚提取率为7.44%。 关键词：中图分类号：TS202.3 文献标识码：A 文章编号：0254-5071（20１5）06-0094-05 doi:     

茶酒糟中茶多酚提取工艺优化及其抗氧化活性的研究

该试验以茶酒糟为原料,乙醇为提取溶剂,采用水浴振荡法提取茶多酚,通过单因素试验探究乙醇体积分数、料液比、提取时间、温度对茶多酚提取量的影响,在此基础上,通过正交试验优化茶多酚的水浴振荡乙醇提取工艺,并考察茶多酚对羟自由基（·OH）、1,1-二苯基-2-三硝基苯肼自由基（DPPH·）的清除能力。结果表明,从茶酒糟中提取茶多酚的最佳提取工艺是：乙醇体积分数60%,料液比1∶30（g∶mL）,提取时间12 min,提取温度75 ℃。在此优化条件下,茶多酚的提取量为41.52 mg/g。从茶酒糟提取得到的茶多酚对·OH和DPPH·最大清除率分别达到91.7%、93.7%,表明茶多酚具有一定的清除自由基的能力,具有较强的抗氧化活性。     

Comparison of the relative dissipation rates of endosulfan pesticide residues between oolong and green tea

The dissipation behaviour of endosulfan in dry made-tea leaves of oolong and green tea was compared to establish whether there was any difference in dissipation rates between the two teas. The dissipation of endosulfan in oolong and green tea corresponded with a first-order kinetics curve. The average half-life of endosulfan (n = 12) was 1.60 ± 0.44 days in green tea and 2.01 ± 0.55 days in oolong tea, showing a statistically significant difference, and indicating that the dissipation of the pesticide was significantly slower in oolong tea than that in green tea. Although the initial levels of residual endosulfan were lower in oolong tea, due to the slower dissipation rate, the residues 5–7 days after application were higher in oolong than in green tea. It is suggested that the minimum interval between endosulfan application and tea leaf harvesting is 7 days for green tea and 10 days for oolong tea in the case where the maximum residue limit of endosulfan in made-tea is fixed as 10 mg kg^?1.     

Effects of phosphorus supply on the quality of green tea

Self-rooted, 10-month-old, uniform tea [Camellia sinensis (L.) O. Kuntze cv. Huangguanyin] plants were supplied with 0, 40, 80, 160, 400 or 1000 μM phosphorus (P) for 17 weeks to determine how P-deficiency affects the quality of green tea. Leaf P concentration increased with increasing P supply. Whole plant dry weight (DW) increased as P supply increased from 0 to 160 μM, then remained little changed with further increasing P supply. The P-deficient green tea displayed decreased concentrations of water extract, total polyphenols, flavonoids, total free amino acids, theanine (Thea) and asparagic acid (Asp) + glutamic acid (Glu), increased concentrations of water soluble sugars, valine (Val), γ-aminobutyric acid (GABA), proline (Pro) and cysteine (Cys), and ratio of total polyphenols to total free amino acids, but unchanged concentrations of total catechins and epigallocatechin gallate (EGCG). In conclusion, the sensory and biochemical qualities of green tea were lowered by P-deficiency.     

The phenolic profiles and antioxidant activity in different types of tea

Phenolics are suggested to be the major bioactive compounds responsible for the health benefits of tea. Seven types of tea were extracted using boiling water. The extraction kinetics was investigated. To more fully characterise the antioxidant profiles and possible associated health benefits of these tea types, the total water‐soluble solid content, phenolics, flavonoids, and antioxidant and antiproliferative activities were quantified. Green tea leaves (GTL) were found to have the highest phenolic content (128.7 ± 1.7 mg g^?1, P < 0.05), followed by teas of decaffeinated green tea bag (dGTB), green tea bag (GTB), black tea bag (BTB), decaffeinated BTB, black tea leaves and oolong tea leaves (OTL). Among the three leaf teas, GT displayed the greatest antioxidant activity (4850.2 ± 60.7 μmol g^?1, P < 0.05), followed by black tea (BT) and oolong tea (OT). As for the four bag teas, decaffeinated GT possessed the strongest antioxidant activity (4431.1 ± 335.0 μmol g^?1, P < 0.05), followed by GT, BT and decaffeinated BT. The results display that phenolic/flavonoid content well correlated with antioxidant activity. Proliferation of three human cancer cells was significantly inhibited in a dose‐dependent manner after exposure to some tea extracts. This may influence consumers in selecting the type of tea and tea brewing times, exhibiting greater health benefits.