Influence of drying methods and agronomic variables on the chemical composition of mate tea leaves (Ilex paraguariensis A. St.-Hil) obtained from high-pressure CO2 extraction

The main objective of this work is to assess the influence of two drying methods (microwave and vacuum oven) and some agronomic variables (plant fertilization conditions and sunlight intensity) on the characteristics of mate tea (Ilex paraguariensis) leaves extracts obtained from high-pressure carbon dioxide extractions performed in the temperature range from 20 to 40 degrees C and from 100 to 250 bar. Samples of mate were collected in an experiment conducted under agronomic control at Ervateira Bar?o LTDA, Brazil. Chemical distribution of the extracts was evaluated by gas chromatography coupled with a mass spectrometer detector (GC/MS). In addition to extraction variables, results showed that both sample drying methods and agronomic conditions exert a pronounced influence on the extraction yield and on the chemical distribution of the extracts.     

Quantitation of methylxanthinic alkaloids and phenolic compounds in mate (Ilex paraguariensis) and their effects on blood vessel formation in chick embryos

Methylxanthinic alkaloids and phenolic compounds are related to the therapeutic properties of Ilex paraguariensis infusions. Considering the known vascular tropism of xanthines, an aqueous extract (mate) and caffeine were evaluated on blood vessel formation, in connection with the analysis of those secondary metabolites, which was performed in young and mature leaf samples collected in three cultivation systems located in the southern region in Brazil (Santa Catarina State). Samples of young and mature leaves from a monoculture cultivation system (MC) showed the highest content of phenolic compounds (149.68 microg/mL, young leaves; 135.50 microg/mL, mature leaves) and caffeine (young leaves, 148.07 microg/mL; mature leaves, 244.63 microg/mL) as compared to samples from agroforesty (AF) and shaded-native (NT) cultures. Theophylline was not detected in samples by reverse phase-high performance liquid chromatography, and mature leaves showed lower theobromine amounts (11.46 microg/mL). Treatments performed with mate aqueous extract and caffeine (1.03-4.12 microM/disk) in the yolk sac vascular membranes of 2-day-old chick embryos revealed pro-vasculo- and angiogenic properties as well as embryonic growth enhancement. These findings, uncoupled from any detectable embryotoxic effect, suggest a potential therapeutic and/or prophylactic use in cardiovascular disorders for caffeine and related constituents of mate plant extracts, an issue that waits further studies.     

Influence of agronomic variables on the composition of mate tea leaves (Ilex paraguariensis) extracts obtained from CO2 extraction at 30 degrees C and 175 bar

The aim of this work is to assess the influence of light intensity (plants with direct sun exposure and in a controlled light intensity) and age of leaves (6-24 months) on the characteristics of the extracts of mate tea leaves obtained from carbon dioxide at high pressures. Samples of mate were collected in an experiment conducted under agronomic control at Industria e Comercio de Erva-Mate Bar?o LTDA, Brazil. The content of selected organic compounds of the extracts was evaluated by gas chromatography together with mass spectrometry. Quantitative analysis of caffeine, theobromine, phytol, vitamin E, squalene, and stigmasterol was performed, and the results showed that field variables exert a strong influence on the liquid yield and on the chemical distribution of the extracts.     

Elemental characterization of commercial mate tea leaves (Ilex paraguariensis A. St.-Hil.) before and after hot water infusion using ion beam techniques

Ilex paraguariensis A. St.-Hil. is used to prepare a traditional tealike beverage widely appreciated in Argentina, Paraguay, Uruguay, and southern Brazil. In these countries, the tea is popularly known as mate or chimarr?o. The aim of this work is to characterize the elemental composition of commercial Ilex paraguariensis and determine the portion of each element present in the leaves that is eluted in the water during the infusion process and consequently ingested by the drinker. Using the particle-induced X-ray emission technique, we verified the presence of Mg, Al, Si, P, S, Cl, K, Ca, Ti, Mn, Fe, Cu, Zn, and Rb at different concentrations, which accounts for about 3.4% of the total mass. The results show a loss of about 90% of K and Cl, 50% of Mg and P, and 20% of Mn, Fe, Cu, Zn, and Rb by the leaves after the infusion. The volume of water used in the infusion affects only the concentration of elements such as Cl, P, K, and Mg until the first 600 mL of water, where a steep decrease in the concentration of these elements was observed in brewed leaves. Furthermore, higher water temperatures (typical temperatures used in infusions, between 80 and 100 degrees C) favor the extraction of K and Cl into the infusion, while the concentration of other elements remains practically constant as a function of temperature.     

Chemical composition and antioxidant activity of yerba-mate (Ilex paraguariensis A.St.-Hil., Aquifoliaceae) extract as obtained by spray drying

Yerba-mate or mate? (Ilex paraguariensis A.St.-Hil., Aquifoliaceae) leaves are typically used for their stimulant, antioxidant, antimicrobial, and diuretic activity, presenting as principal components polyphenolic compounds. In this study, the objective was to develop a yerba-mate dry extract by using spray drying technology and to evaluate the dry extract antioxidant activity and chemical composition. The results obtained by means of the DPPH assay show that the extract presents an IC(50) of 2.52 mg/mL. The yerba-mate spray-dried extract presents high catalase-like activity, suggesting that it is a strong free-radical scavenger. The antioxidant activity as expressed as catalase-like activity was related to total polyphenol content. In addition, the results show that the spray-dried extract presents high polyphenol content, namely, high concentrations of caffeic acid (1.54 mg/g), 5-caffeoylquinic acid (91.40 mg/g), rutin (5.38 mg/g), and total phenolics (178.32 mg/g), which justifies its high antioxidant activity.     

Hydrolysis influence on phytochemical composition, antioxidant activity, plasma concentration, and tissue distribution of hydroethanolic Ilex paraguariensis extract components

The infusion of aerial parts of Ilex paraguariensis is widely consumed. Its antioxidant activity suggests an important role of this plant in the treatment/prevention of oxidative stress related diseases. Plant extract active compounds are frequently found in esterified form that may be poorly absorbed. Hydrolysis of the extract is a possible approach to increase its bioavailability. The aim of this study was to perform a phytochemical analysis and evaluate in rats the plasma concentration and tissue distribution of antioxidant compounds in the hydroethanolic extract of Ilex paraguariensis, before and after enzymatic hydrolysis. Both extracts presented high antioxidant activity and phenolic content. Rats given single or repeated doses of the hydrolyzed extract showed increased plasma antioxidant activity and higher plasma levels of caffeic acid. However, no changes of endogenous antioxidants were observed. In conclusion, hydrolysis of the extract of Ilex paraguariensis is a strategy to improve its bioavailability and in vivo antioxidant activity.     

Selective liquid CO2 extraction of purine alkaloids in different Ilex paraguariensis progenies grown under environmental influences

In this investigation, liquid carbon dioxide at 20 degrees C and 150 bar was used for the selective extraction of caffeine and theobromine from dry leaves of mate. A comparison between the chromatograms from CO2 extraction and traditional solvent extraction supports the selectivity of carbon dioxide for these purine alkaloids. The advantages of selective liquid CO2 extraction in terms of speed and resolution of UV/HPLC is also evidenced. A randomized block design of experiments was proposed to investigate the influence of 16 progenies of Ilex paraguariensis grown in 3 diverse sites on the contents of caffeine and theobromine in liquors of mate leaves obtained by extraction with compressed CO2. A significant effect of both these factors on the parameters investigated was observed by involving the F distribution in the statistical analysis. A cluster analysis based on the experimental uncertainties in the contents of these two methylxanthines has identified from four to six different groups of mate progenies.     

Polyphenolic compounds, antioxidant capacity, and quinone reductase activity of an aqueous extract of Ardisia compressa in comparison to mate (Ilex paraguariensis) and green (Camellia sinensis) teas

Aqueous extracts of the leaves of Ardisia compressa (AC) have been used in folk medicine to treat various liver disorders including liver cancer. The objective of this study was to partially characterize and determine the total polyphenol content, antioxidant capacity, and quinone reductase activity of A. compressa tea in comparison to mate (Ilex paraguariensis, MT) and green (Camellia sinensis,GT) teas. Total polyphenol content, antioxidant capacity, and phase II enzyme induction capacity were measured by the modified Folin-Ciocalteu, ORAC, and quinone reductase (QR) assays, respectively. The major polyphenols in AC were not catechins. HPLC retention times and standard spikes of AC indicated the presence of gallic acid, epicatechin gallate, ardisin and kaempferol. Using catechin as standard, the total polyphenol value of AC (36.8 +/- 1.1 mg/mg DL) was significantly lower than GT (137.2 +/- 5.8 mg equivalent of (+)-catechin/mg dried leaves, DL) and MT (82.1 +/- 3.8 mg/mg DL) (P < 0.001). Antioxidant capacity (AC, 333; GT, 1346; MT, 1239 mmol Trolox equivalents/g DL) correlated with total polyphenol values (r(2) = 0.86, P < 0.01). AC (4.5-12.5 microg/mL) induced QR enzyme, in Hepa1c1c7 cells, up to 15%. MT and GT showed no induction at the concentrations tested (0.5-10.5 and 0.5-12.5 mg/mL, respectively). These results suggest that AC has a different mechanism of protection against cytotoxicity that is not related to its antioxidant capacity. Further studies are needed to determine such mechanisms and to explore its potential as a chemopreventive or therapeutic agent.     

Effect of yerba mate (Ilex paraguariensis) tea on topoisomerase inhibition and oral carcinoma cell proliferation

Tea flavonoids have antitopoisomerase activity and can inhibit cell proliferation. The objectives of this study were to determine the phenolic content of yerba mate tea products (MT) (Ilex paraguariensis) and evaluate their capacity to inhibit topoisomerase I (Topo I) and II (Topo II) activities and oral carcinoma cell proliferation. Total polyphenols of aqueous extracts of dried MT leaves were measured by the Folin-Ciocalteau assay, using chlorogenic (CH) and gallic (GA) acids as standards. Topoisomerase inhibition was determined by a clone-forming assay, which uses yeast (Saccharomyces cerevisiae) strains as a model. Controls included dimethyl sulfoxide (1.66%); camptothecin (50 microg/mL), a Topo I inhibitor; and amsacrine (100 microg/mL), a Topo II inhibitor. Cytotoxicity studies were conducted using a nontumorigenic human keratinocyte cell line HaCaT and two human squamous cancer cell lines (SCC-61 and OSCC-3). MT was found to be a rich source of phenolic compounds. Total polyphenol content of various commercially available traditional MT products ranged from 236 to 490 mg equiv of CH/g of dry leaves. Such levels were significantly different among products depending on their origin (P < 0.001). Higher anti-topoisomerase II activity was observed against JN394t(2-4) strain for Nobleza Gaucha MT (IC50 = 0.43 microg equiv of CH) in comparison to GA (IC50 = 112 mM) and CH (IC50 > 1500 mM). MT showed catalytic anti-topoisomerase activity against Topo II but not against Topo I. In addititon, MT exhibited dose-dependent cytotoxicity against all squamous cell lines tested. In comparison to premalignant cell line HaCaT [28 microg equiv of (+)-catechin mL(-1)], the cell line SCC-61 [21 microg equiv of (+)-catechin mL(-1)] was the most sensitive to MT, resulting in 50% inhibition of net cell growth. It is concluded that MT is rich in phenolic constituents and can also inhibit oral cancer proliferation. The effect on cancer cell proliferation may be mediated via inhibition of topoisomerase II. The lack of correlation between polyphenol content and the inhibition of topoisomerases suggests that the effect of MT on topoisomerase inhibition may be due to other still unidentified biologically active phytochemicals constituents.     

Effect of growing and drying conditions on the phenolic composition of mate teas (Ilex paraguariensis)

Mate tea ( Ilex paraguariensis) has been used for centuries and is widely consumed in Brazil, Argentina, Paraguay, and Uruguay. The aim of the present study was to determine how growing and drying conditions affect the phenolic concentration and antioxidant capacity of 15 Mate teas from forest or plantation cultivations, dried either with hot air or wood smoke. The total polyphenol concentration determined with Folin-Ciocalteu ranged from 100.3 +/- 5.5 to 179.7 +/- 3.6 mg equiv chlorogenic acid/g dry leaves. The antioxidant capacity according to the oxygen radical absorbance capacity assay ranged from 1.5 +/- 0.3 to 4.1 +/- 0.1 mmol Trolox equiv/g dry leaves. Ten phenolic compounds were identified and correlated with antioxidant capacity ( R (2) = 0.80). Principle component analysis and multivariate linear regression were conducted to assess the effect of growing and drying conditions. Sun-exposed (plantation grown) Mate teas exhibited higher levels of all polyphenols as compared to shaded (forest grown) Mate teas ( P < 0.05). Lower rainfall, temperature, and drying conditions had varying effects on the phenolics. On average, plantation grown Mate teas had a greater concentration of phenolics than forest grown teas. Thus, plantation grown Mate teas represent better potential sources for their commercial extraction.     

Classification of Ilex species based on metabolomic fingerprinting using nuclear magnetic resonance and multivariate data analysis

The metabolomic analysis of 11 Ilex species, I. argentina, I. brasiliensis, I. brevicuspis, I. dumosavar. dumosa, I. dumosa var. guaranina, I. integerrima, I. microdonta, I. paraguariensis var. paraguariensis, I. pseudobuxus, I. taubertiana, and I. theezans, was carried out by NMR spectroscopy and multivariate data analysis. The analysis using principal component analysis and classification of the (1)H NMR spectra showed a clear discrimination of those samples based on the metabolites present in the organic and aqueous fractions. The major metabolites that contribute to the discrimination are arbutin, caffeine, phenylpropanoids, and theobromine. Among those metabolites, arbutin, which has not been reported yet as a constituent of Ilex species, was found to be a biomarker for I. argentina,I. brasiliensis, I. brevicuspis, I. integerrima, I. microdonta, I. pseudobuxus, I. taubertiana, and I. theezans. This reliable method based on the determination of a large number of metabolites makes the chemotaxonomical analysis of Ilex species possible.     

Extraction of purine alkaloids from maté (Ilex paraguariensis) using supercritical CO(2).

Experimental data for the supercritical CO(2) extraction of purine alkaloids (caffeine, theobromine, and theophylline) from ground herbal maté tea (Ilex paraguaryensis) using a high-pressure apparatus are presented. Caffeine, theophylline, and theobromine were identified in the extracted fractions using HPLC. Results indicated a much higher CO(2) selectivity for caffeine in comparison with those for theophylline and theobromine. Solubilities of pure compounds in carbon dioxide were also determined at 313.2, 323.2, 338.2, and 343.2 K, and pressures ranging from 14 to 24 MPa. Caffeine solubility exhibited a retrograde behavior with temperature while theophylline and theobromine manifested a normal behavior at conditions explored in this study. Solubilities in binary CO(2)/purine alkaloid model systems were much higher than those obtained during extraction of maté tea, demonstrating the difficulty of using binary data in predicting complex multicomponent behavior.     

Polyphenol composition of a functional fermented tea obtained by tea-rolling processing of green tea and loquat leaves

Phenolic constituents of a new functional fermented tea produced by tea-rolling processing of a mixture (9:1) of tea leaves and loquat leaves were examined in detail. The similarity of the phenolic composition to that of black tea was indicated by high-performance liquid chromatography comparison with other tea products. Twenty-five compounds, including three new catechin oxidation products, were isolated, and the structures of the new compounds were determined to be (2R)-2-hydroxy-3-(2,4,6-trihydroxyphenyl)-1-(3,4,5-trihydroxyphenyl)-1-propanone 2-O-gallate, dehydrotheasinensin H, and acetonyl theacitrin A by spectroscopic methods. In addition, theacitrinin A and theasinensin H were obtained for the first time from commercial tea products. Isolation of these new and known compounds confirms that reactions previously demonstrated by in vitro model experiments actually occur when fresh tea leaves are mechanically distorted and bruised during the production process.     

Characterization and phenanthrene sorption of tea leaf powders

The sorption of hydrophobic organic compounds by natural organic matter is mainly regulated by its aromatic and aliphatic fractions, but it is not clear which fraction is more dominant. In this study, six types of Chinese tea leaves (three varieties of tender and three corresponding mature leaves) and their respective brewed ones were analyzed by elemental analysis, 13C NMR, and Fourier transform infrared attenuated total reflectance. Their sorption of phenanthrene was examined using a batch equilibration technique. The aromatic carbon content of tea leaves reduced while the aliphatic carbon content increased with increasing maturity and brewing. Sorption isotherms by all of the 12 tea sorbents were practically linear, showing a partition type sorption. The phenanthrene KOC (organic carbon-normalized sorption capacity) of the 12 sorbents ranged from 6960 to 32,900 mL/g, which increased with increasing aliphatic carbons and decreasing aromatic carbons in the tea leaves. The dissolved organic matter was released into solution from the sorbents during the sorption process, which could bind phenanthrene in the solution, and thereafter leading to underestimation of sorption capacity of the sorbents, but this did not change the correlation trends between KOC and functional carbon group content. Aliphatic fractions rather than aromatic moieties regulated the phenanthrene sorption of the tea leaf powders used in this study.     

凤凰单枞茶树吸收土壤铬和锰的规律研究

[目的]探究凤凰单枞茶树对土壤中铬和锰的吸收规律,为综合防治茶园土壤Cr和Mn污染提供科学依据。[方法]对粤东揭阳市揭东县宝山茶园凤凰单枞茶树和土壤进行采样,运用原子吸收分光光度计法测定茶叶、茶茎和茶根中Cr和Mn的含量,并采用乙酸提取法和原子吸收分光光度计法测定土壤样品中酸可提取态Cr和Mn的含量。[结果]茶树各器官Cr含量由高到低分布情况依次为:茶根>茶茎>茶叶;而Mn含量分布情况依次为:茶叶>茶茎>茶根。茶叶中Cr和Mn含量与土壤中酸可提取态Cr和Mn呈显著正相关性,茶茎中Cr含量与土壤中酸可提取态Cr也呈显著正相关性;茶叶、茶茎和茶根中的Cr含量均与土壤中有机质含量呈显著正相关性,茶根中的Mn含量与土壤pH值呈显著负相关性,茶叶中的Mn与土壤有机质呈显著正相关性。[结论]通过调节土壤酸可溶解态Cr和Mn,土壤pH值和有机质含量,从而控制茶树吸收Cr和Mn的量,可以达到防治茶叶重金属污染,提高茶叶质量的目的。     

茶园施肥模式对茶叶黄酮类及糖苷类代谢物含量的影响

【目的】研究施肥模式对茶叶生化成分含量和黄酮类及糖苷类代谢物累积的影响,为实现茶叶优质高产提供科学依据。【方法】田间试验设在四川茶树种植典型区域—雅安,试验连续进行了3年,设置不施肥(CK)、常量化肥(N 585 kg/hm2,TF)、有机肥替代25%化肥氮(OF)、减施25%氮肥(SF) 4个处理。分析了不同处理茶叶主要品质成分含量,并基于非靶向代谢组学检测结果分析了黄酮类及糖苷类代谢物累积差异。【结果】与C K处理相比,T F、 O F、 S F处理茶氨酸、游离氨基酸含量分别提高了7.2 2%～1 3.4 0%、23.15%～25.50%,可溶性糖和茶多酚含量分别降低了8.17%～13.86%、6.08%～11.49%,表儿茶素、表没食子儿茶素、芸香苷等10种代谢物累积水平显著下降。与TF处理相比,SF处理茶氨酸、咖啡碱和水浸出物含量分别降低了5.45%、5.97%、8.91%,茶多酚、可溶性糖、游离氨基酸含量和酚氨比无显著差异;芍药素-3-O-葡萄糖苷、肉桂萜醇C1-19-葡萄糖苷、查耳酮樱花素等7种代谢物累积水平显著降低,川陈皮素、牡荆素-4’-O-α-L-吡喃鼠李糖苷、(S)...     

Application of near-infrared reflectance spectroscopy to the simultaneous prediction of alkaloids and phenolic substances in green tea leaves

A near-infrared reflectance spectroscopic (NIRS) method for the prediction of polyphenol and alkaloid compounds in the leaves of green tea [Camellia sinensis (L.) O. Kuntze] was developed. Reference measurements of the individual catechins, gallic acid, caffeine, and theobromine were performed by reversed-phase HPLC. The total polyphenols were determined according to the colorimetric Folin-Ciocalteu assay. Using the partial least-squares algorithm, very good calibration statistics were obtained for the prediction of gallic acid, (-)-epicatechin, (-)-epigallocatechin, (-)-epicatechin gallate, (-)-epigallocatechin gallate, caffeine, and theobromine (R(2) > 0.85) with standard deviation/standard error of cross-validation (SD/SECV) ratio ranging from 2.00 to 6.27. Simultaneously, the dry matter content of the tea leaves can be analyzed very precisely (R(2) = 0.94; SD/SECV = 4.12). Furthermore, it is possible to discriminate tea leaves of different age by principal component analysis on the basis of the received NIR spectra. Prediction of the total polyphenol content is performed with a lower accuracy, which might be due to the lack of specificity in the colorimetric reference method. The study demonstrates that NIRS technology can be successfully applied as a rapid method not only for breeding and cultivation purposes but also to estimate the quality and taste of green tea and to control industrial processes, for example, decaffeination.     

采用PLS-DA分析毛火方式对工夫红茶品质的影响

为探究不同毛火方式对工夫红茶品质的影响,明确新型电磁内热式滚筒-热风耦合干燥设备的毛火效果,该研究以一芽一二叶初展嫩度的"福鼎大白"品种为原料进行工夫红茶加工,设定电磁滚筒-热风耦合(Rotary pot-Hot air coupling First-Drying with electromagnetic heat,RHFD)、链板热风(Chain plate Hot air First-Drying,CHFD)、箱式热风(Box Hot air First-Drying,BHFD)、滚筒式滚炒(Rotary pot First-Drying,RFD)等4种毛火方式,比较所制茶样的茶多酚、儿茶素、茶色素、可溶性糖、咖啡碱、氨基酸等29个非挥发性指标,114个气相色谱-质谱技术(GasChromatography-Mass Spectrometry,GC-MS)检测的挥发性香气指标,10个外形和汤色色泽客观评价指标,同时进行了毛火方式的热效率、生产效率、生产成本等性能指标的分析比较,通过偏最小二乘判别统计(PartialLeastSquaresDiscriminationAnalysis,PLS-DA)分析毛火方式对优质工夫红茶品质的影响,并获得标志性差异化合物。结果表明:电磁内热式滚筒-热风耦合毛火处理下茶多酚和儿茶素总量显著最低(P<0.05),简单儿茶素含量较高,茶红素和可溶性糖含量、茶黄素综合指标TDE和茶色素综合指标10TFRB最高(P<0.05),毛火方式对茶黄素总量影响不显著(P>0.05);挥发性化合物总量以RHFD方式最高,RFD方式次之,CHFD方式最低;RHFD毛火方式芳香类、萜烯类等化合物含量最高。电磁内热式滚筒-热风耦合毛火升温快、温度分布均匀且稳定性好,热效率和生产效率高(分别为50.0%、220 kg/h),生产成本较低(仅0.32元/kg),预热时间仅14min;所制红茶在汤色透亮度、香气甜久度、滋味甜醇度等方面均得到提升,感官总分最高(P<0.05),达88.1。PLS-DA分析从挥发性和非挥发性角度均可将工夫红茶4种毛火方式显著区分,并分别获得了43种和18种差异化合物,结合差异性分析获得标志性差异化合物,2,4,6-三(1,1-二甲基乙基)-4-甲基环己-2,5-二烯-1-酮、香叶醇、3-辛酮、水杨酸甲酯、茶黄素、茶褐素、可溶性糖、表儿茶素等,可作为区分工夫红茶毛火方式,以及定向加工甜香、甜醇、高亮等优质工夫红茶的指标物质。该研究为红茶加工基础和品质提升提供技术参考和理论指导。     

Effect of microbial fermentation on content of statin, GABA, and polyphenols in Pu-Erh tea

Besides cancer prevention, the hypolipidemic effects of tea have been well studied in animals and humans. Recently, statin has been identified in Pu-erh tea extract. Clinical trials have confirmed that statin decreases the incidence of major coronary and cerebrovascular events and this may be due to its hypolipidemic and antiinflammatory effects. Since a good Pu-erh tea needs longer storage (10 years or more) of fermentation to enhance the flavor and fragrance, we screened microorganisms from two Pu-erh teas, 20 and 25 years old. Species of fungi and bacteria strains that contributed to a good taste of Pu-erh tea were isolated. The effect of fermentation was investigated by inoculating fresh tea leaves with individual strains of isolated microorganisms. Results showed that statin, total polyphenol content, and the scavenging activities of alpha,alpha-diphenyl-beta-picrylhydrazyl (DPPH) radicals increased during fermentation. Tea leaves inoculated with Streptomyces bacillaris strain R9 had the highest polyphenol content (3.3 mg/100 g) and scavenging ability to DPPH radicals (92%). Streptomyces cinereus strain Y11 was equally good for polyphenol content but yielded the highest amount of statin (1012 ng/g) after 42 days of fermentation. Interestingly, the statin content of fresh tea leaves fermented with strain R9 or Y11 after 180 days was much higher (4- and 8-fold, respectively) than that of the 25-year-old Pu-erh tea (513 ng/g) as measured by the HPLC method. Similarly, these two strains also increased the content of gamma-aminobutyric acid (GABA) 5.7- and 4.7-fold in tea fermented for 180 days as compared with the fresh leaves (1270 microg/g) and that were higher than that of the Pu-erh tea (4900 microg/g). Taken together, the present results indicate that tea short-term fermented with S. bacillaris or S. cinereus enhances the color and content of statin, GABA, and polyphenols.     

Structure-activity relationships of tea compounds against human cancer cells

The content of the biologically active amino acid theanine in 15 commercial black, green, specialty, and herbal tea leaves was determined as the 2,4-dinitrophenyltheanine derivative (DNP-theanine) by a validated HPLC method. To define relative anticarcinogenic potencies of tea compounds and teas, nine green tea catechins, three black tea theaflavins, and theanine as well as aqueous and 80% ethanol/water extracts of the same tea leaves were evaluated for their ability to induce cell death in human cancer and normal cells using a tetrazolium microculture (MTT) assay. Compared to untreated controls, most catechins, theaflavins, theanine, and all tea extracts reduced the numbers of the following human cancer cell lines: breast (MCF-7), colon (HT-29), hepatoma (liver) (HepG2), and prostate (PC-3) as well as normal human liver cells (Chang). The growth of normal human lung (HEL299) cells was not inhibited. The destruction of cancer cells was also observed visually by reverse phase microscopy. Statistical analysis of the data showed that (a) the anticarcinogenic effects of tea compounds and of tea leaf extracts varied widely and were concentration dependent over the ranges from 50 to 400 microg/mL of tea compound and from 50 to 400 microg/g of tea solids; (b) the different cancer cells varied in their susceptibilities to destruction; (c) 80% ethanol/water extracts with higher levels of flavonoids determined by HPLC were in most cases more active than the corresponding water extracts; and (d) flavonoid levels of the teas did not directly correlate with anticarcinogenic activities. The findings extend related observations on the anticarcinogenic potential of tea ingredients and suggest that consumers may benefit more by drinking both green and black teas.