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.     

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.     

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.     

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.     

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.     

不同品种乌龙茶种植后土壤肥力和茶叶品质的变化

本研究以毛蟹(C.sinensis cv.Maoxie)、本山(C.sinensis cv.Benshan)、梅占(C.sinensis cv.Meizhan)3个乌龙茶品种为研究对象,分析不施肥条件下茶树根部土壤养分随种植年限的变化,以及茶叶品质的变化。结果表明,不同茶树的根部土壤p H值均随茶树种植年限的增长而呈现下降趋势。不同茶树根部土壤的有机质及有效磷含量达到Ⅰ级肥力;0年取样的茶树土壤碱解氮含量为Ⅰ级肥力,不施肥种植1~2年后均为Ⅱ级肥力;土壤速效钾含量在茶树0年取样时为Ⅰ级肥力,不施肥种植1年后毛蟹根部土壤为Ⅲ级肥力,本山和梅占则为Ⅱ级肥力,2年后均为Ⅲ级肥力。茶树叶片品质分析结果表明,随着不施肥种植时间的延长,茶叶中茶多酚、茶氨酸、咖啡碱的含量呈现下降趋势。同一种茶树在不同种植年限间,茶多酚含量的变化较为微弱,而茶氨酸、咖啡碱含量则呈显著差异。不同种植年限下茶树叶片中表没食子儿茶素没食子酸酯、表儿茶素没食子酸酯与表没食子儿茶素含量及其品质指数均随着不施肥种植时间的延长而呈下降趋势,且同一种茶树不同种植年限间的变化率均呈显著差异。     

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.     

Nitrogen and potassium effects on the macronutrient and micronutrient content of zoysiagrasses 1

Zoysiagrass has more potential for utilization in the Gulf Coast states. There has been minimal research on the nitrogen (N) and potassium (K) fertility response of zoysiagrass and the resulting effect they have on the macronutrient and micronutrient content. The objective of this study was to evaluate the effects of N and K fertility on the nutrient content of zoysiagrass. A study was conducted on four zoysiagrasses: Zoysiajaponica x Z. tenuifolia Willd. ex Trin. ('Emerald'); Z. japonica Steud. ('El Toro’ and ‘Meyer'); and Z. matrella. The study was a completely randomized design with 3 replications. It was a 2x2 factorial with the factors being N and K at two levels that were imposed on the four zoysiagrass cultivars. The N and K treatment combinations consisted of high (H) and low (L) rates of N and K at the following levels: N levels of 454 and 227 g N 93 m^‐2 month^‐1 and K levels of 454 and 227 g K 93 m^‐2month^‐1. The treatment combinations were (N and K): HH, HL, LH, and LL and were applied in 2 split applications monthly from July through November. All plots received two applications of a micronutrient fertilizer (June and August), were irrigated as needed, and maintained at a height of 3.8 cm weekly. Plant tissue samples were collected in September and analyzed for nitrogen (N), phosphorus (P), K, calcium (Ca), magnesium (Mg), sulfur (S), boron (B), copper (Cu), iron (Fe), manganese (Mn), and zinc (Zn) concentrations. There were significant differences for the concentrations of N, K, P, Ca, Mg, B, Cu, Fe, Mn, and Zn. The concentrations of K, Ca, and Mg were below the sufficiency range for these nutrients for all cultivars and treatments.     

Influence of agronomic factors and extraction rate on the acrylamide contents in yeast-leavened breads

Because the impact of agronomical factors on bakery products quality is still an insufficiently studied field, acrylamide contents of breads produced from flours of nine wheat, two rye, and two spelt varieties harvested in 2003 and 2004 were investigated. It could be demonstrated that acrylamide content in bread strongly depends on the cultivar, with extremes differing by a factor of 5.4 due to marked differences in free asparagine and crude protein contents. Nitrogen fertilization also resulted in elevated amino acid and protein contents, thus increasing acrylamide levels from 10.6 to 55.6 mug/kg. Independent of fertilization, harvest year turned out to be another factor influencing acrylamide formation. Breads produced from 2003 flours showed significantly higher acrylamide contents than those of 2004, which was ascribed to favorable light and temperature conditions during the cultivation period, thus enhancing amino acid and protein contents. Sprouting of the grain also resulted in significantly higher acrylamide levels, which was attributed to elevated enzyme activities and the formation of precursors from protein and starch. Furthermore, bakery products made from flours with higher extraction rates were shown to contain higher acrylamide levels resulting from extracted free asparagine and protein from the aleuron layers of the cereal grain.     

Anti-tobacco mosaic virus (TMV) triterpenoid saponins from the leaves of Ilex oblonga

Ten triterpene saponins have been isolated from a MeOH extract of the leaves of Ilex oblonga. In their structures, six new triterpenoid saponins were named as oblonganosides H-M (2, 4, 5, and 8-10). All structures were elucidated on the basis of spectroscopic analysis. Among the triterpenoid saponins, three compounds (7, 8, and 10) showed obvious activities in inhibiting multiplication of the tobacco mosaic virus (TMV).     

水稻生物化学参数与高光谱遥感特征参数的相关分析

通过不同氮素营养水平的水稻田间试验，研究稻叶生物化学成份(叶绿素、类胡萝卜素、蛋白质、纤维素、淀粉含量) 与水稻冠层高光谱遥感特征参数之间的关系，结果表明，随着叶位的下移，叶片叶绿素含量与光谱特征变量之间的相关性明显减弱，绿峰反射率、红谷反射率、红边波长、蓝边面积和“三边”面积构成的植被指数等变量与上叶叶绿素含量、类胡萝卜素、纤维素、叶鞘淀粉含量之间有较好的相关性，而蛋白质含量与各种变量的相关系数均非常低。兰边面积、兰边面积和红边面积构成的植被植数与上叶叶绿素含量之间，红边面积 、绿反射峰与红反射峰构成的植被指数与叶鞘淀粉含量之间的相关系数都达到了0.01极显著检验水平，因此，可利用这些变量建立上叶叶绿素a、纤维素和叶鞘淀粉含量的估测模型。     

Analysis of glycosidically bound aroma precursors in tea leaves. 2. Changes in glycoside contents and glycosidase activities in tea leaves during the black tea manufacturing process

Glycosides are known to be precursors of the alcoholic aroma compounds of black tea. They are hydrolyzed by endogenous glycosidases during the manufacturing process. Changes in the amounts of these glycosides during the manufacturing process were investigated by using a capillary gas chromatographic--mass spectrometric analysis after trifluoroacetyl derivatization of the tea glycosidic fractions. Primeverosides were 3-fold more abundant than glucosides in fresh leaves, but they decreased greatly during the manufacturing process, especially during the stage of rolling. After the final stage of fermentation, primeverosides had almost disappeared, whereas glucosides were substantially unchanged. These results show that hydrolysis of the glycosides mainly occurred during the stage of rolling and confirm that primeverosides are the main black tea aroma precursors. This was also supported by the changes in the glycosidase activities in tea leaves. The glycosidase activities remained at a high level during withering but decreased drastically after rolling.     

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.     

喷施稀土对辣椒体内稀土元素含量和分布的影响

利用ＩＣＰ－ＭＳ测定了辣椒不同部位１５种单一稀土元素的含量。结果表明，喷施稀土的球椒植株，各部分稀土含量的高低顺序为叶〉根〉茎〉果，而对照的顺序为根〉茎〉果；随着喷施长稀土浓度的增大辣椒各部位稀土含量有上升的趋势，当喷施浓度为６００ｍｇ／Ｌ，各部位稀土含量最大值。     

氟对茶树生长、叶片营养元素含量、儿茶素类物质和香气成分的影响

本试验为水培试验,研究了不同氟浓度处理(0.1、 0.2、 0.3 mmol/L F)对茶苗(平阳特早)干物质量、 相对生长量、 叶片营养元素含量、 酚类物质和香气成分的影响。结果表明,随着氟浓度的增加,茶苗的干物质积累和相对生长率显著降低。氟处理显著增加了茶苗叶片总磷和氟的含量,显著降低了总氮与碳酸氢根的含量,但对叶片氯离子含量的影响不显著。随着氟浓度的增加,叶片钾、 镁、 铁、 锌的含量显著降低,而钙含量显著增加。对茶叶儿茶素类物质而言,氟处理显著降低了茶苗叶片四种儿茶素（EGC、 EC、 EGCG、 ECG）和茶氨酸、 咖啡碱的含量。此外, 0.1 mmol/L F处理降低了茶苗叶片中醛、 醇、 酯和芳香类物质含量,而0.2 mmol/L F和0.3 mmol/L F处理增加了茶苗叶片主要的香气成分。     

Varietal and interspecific influence on micronutrient contents in citrus from the Mediterranean area

To specify the genotypic variation of Mediterranean Citrus juices, the contents of carotenoids, flavonoids, and vitamin C were determined by high-performance liquid chromatography. A selection of orange varieties and Mandarin species from the Mediterranean area (Citrus sinensis, Citrus deliciosa Ten, and Citrus clementina Hort. ex Tan) was evaluated using carotenoid profiles and flavanones contents. Among the eight varieties of orange (Salustiana, Hamlin, Shamouti, Pera, Valencia, Maltaise, Sanguinelli, and Cara-cara) and two Mandarin species, only three cultivars (Pera, Sanguinelli, and Shamouti) and the two Mandarin species displayed a high content of vitamin A (374, 381, and 272 ER L(-1) for the three orange cultivars and 1156 and 960 retinol equivalent (RE) L(-1) for the Mandarins) due to a high content of beta-cryptoxanthin. These same Citrus were also rich in hesperidin (502, 537, 552, 767, and 754 mg L(-1), respectively). Principal component analysis allowed the Mediterranean orange varieties and Mandarin species to be differentiated on the basis of nutritional criteria. Strong correlations were observed between beta-cryptoxanthin and hesperidin (r = 0.92) and between beta-cryptoxanthin and beta-carotene (r = 0.98). In contrast, vitamin C content was not correlated with carotenoids and flavanone glycosides. The Mandarin and orange group was quite distinct. The orange varieties could be divided in two groups. In addition, a diversity tree allowed a genetic approach to differentiating Citrus cultivars on the basis of Euclidian distances. This representation showed that the hybrid Clementine was nearer to its parent Mandarin than to its parent orange, suggesting that beta-cryptoxanthin was a dominant genetic factor. With regard to vitamin A, Mandarin and its hybrid Clementine appeared to be the best Citrus species.     

Factors affecting the levels of tea polyphenols and caffeine in tea leaves

An isocratic HPLC procedure was developed for the simultaneous determination of caffeine and six catechins in tea samples. When 31 commercial teas extracted by boiling water or 75% ethanol were analyzed by HPLC, the levels of (-)-epigallocatechin 3-gallate (EGCG), and total catechins in teas were in the order green tea (old leaves) > green tea (young leaves) and oolong tea > black tea and pu-erh tea. Tea samples extracted by 75% ethanol could yield higher levels of EGCG and total catechins. The contents of caffeine and catechins also have been measured in fresh tea leaves from the Tea Experiment Station in Wen-Shan or Taitung; the old tea leaves contain less caffeine but more EGCG and total catechins than young ones. To compare caffeine and catechins in the same tea but manufactured by different fermentation processes, the level of caffeine in different manufactured teas was in the order black tea > oolong tea > green tea > fresh tea leaf, but the levels of EGCG and total catechins were in the order green tea > oolong tea > fresh tea leaf > black tea. In addition, six commercial tea extracts were used to test the biological functions including hydroxyl radical scavenging, nitric oxide suppressing, and apoptotic effects. The pu-erh tea extracts protected the plasmid DNA from damage by the Fenton reaction as well as the control at a concentration of 100 microg/mL. The nitric oxide suppressing effect of tea extracts was in the order pu-erh tea >/= black tea > green tea > oolong tea. The induction of apoptosis by tea extract has been demonstrated by DNA fragmentation ladder and flow cytometry. It appeared that the ability of tea extracts to induce HL-60 cells apoptosis was in the order green tea > oolong > black tea > pu-erh tea. All tea extracts extracted by 75% ethanol have stronger biological functions than those extracted by boiling water.     

Influence of CaCl(2) on the foliar biomass and quality of tobacco leaves

The aim of the present work was to determine the influence of different CaCl(2) dosages on foliar biomass and quality of tobacco plants (Nicotiana tabacum L. var. Tennesse 86). Plants were grown under controlled conditions and submitted to regular fertilization with macro- and micronutrients. The CaCl(2) was applied to the nutrient solution at 1.25 mM (T1), 2.5 mM (T2), and 5 mM (T3). The results indicated that, under the experimental conditions of this work, the application of 1.25 mM of CaCl(2) favored the growth and development of the leaves, this leading to improved biomass production in tobacco leaves. The increase in foliar biomass in treatment T1 could largely be a result of the stimulation of NO(3)(-) assimilation. However, this situation has negative consequences for tobacco quality, given that the T1 treatment augmented the NO(3)(-) concentration and the foliar concentration of nicotine (both effects being harmful for human consumption) and decreased the concentration of reducing sugars in leaves of tobacco plants compared with those of T2 and T3. Finally, a close and directly proportional relationship was found in our experiment between the parameters of foliar-biomass production, total N concentration, chlorophyll concentration, and decline in quality.