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.     

Influence of agronomic variables on the macronutrient and micronutrient contents and thermal behavior of mate tea leaves (Ilex paraguariensis)

The influence of agronomic variables (light intensity, age of leaves, and fertilization type) on the content of macronutrients and micronutrients (potassium, calcium, sodium, magnesium, manganese, iron, zinc, and copper) of tea leaves was assessed by acid digestion, followed by flame atomic absorption spectrometry (FAAS). The thermal behavior of mate tea leaves (Ilex paraguariensis) was also studied in this work. Samples of mate (Ilex paraguariensis) were collected in an experiment conducted under agronomic control at Erva-Mate Bar?o Commerce and Industry LTD (Brazil). The results showed that the mineral content in mate is affected by the agronomic variables investigated. In general, the content of mineral compounds analyzed is higher for younger leaves and for plants cultivated in shadow. Thermal analysis of samples indicated a similar behavior, with three typical steps of decomposition: loss of water, degradation of low-molecular weight compounds, and degradation of residual materials.     

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.     

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.     

Metabolite profiling of Angelica gigas from different geographical origins using 1H NMR and UPLC-MS analyses

Angelica gigas obtained from different geographical regions was characterized using (1)H nuclear magnetic resonance (NMR) spectroscopy and ultraperformance liquid chromatography-mass spectrometry (UPLC-MS) followed by multivariate data analyses. Principal component analysis (PCA) and orthogonal partial least-squares-discriminant analysis (OPLS-DA) score plots from (1)H NMR and UPLC-MS data sets showed a clear distinction among A. gigas from three different regions in Korea. The major metabolites that contributed to the discrimination factor were primary metabolites including acetate, choline, citrate, 1,3-dimethylurate, fumarate, glucose, histamine, lactose, malate, N-acetylglutamate, succinate, and valine and secondary metabolites including decursin, decursinol, nodakenin, marmesin, 7-hydroxy-6-(2R-hydroxy-3-methylbut-3-ethyl)coumarin in A. gigas roots. The results demonstrate that (1)H NMR and UPLC-MS-based metabolic profiling coupled with chemometric analysis can be used to discriminate the geographical origins of various herbal medicines and to identify primary and secondary metabolites responsible for discrimination.     

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.     

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.     

Hybridization barriers among the species of Arachis L., namely of the sections Arachis (including the groundnut) and Erectoides

Some members of section Erectoides have been found resistant to early leaf spot disease, a serious constraint to groundnut productivity. These accessions do not cross with cultivated species. Crossing between one such diploid species, A. paraguariensis (ICGs 8130 and 8973) of section Erectoides, and the diploids, A. batizocoi and A. duranensis, and the tetraploid A. hypogaea (groundnut) of section Arachis has helped understand barriers to hybridization between sections. These crosses result in the development of normal pegs and pods, but with of ovule and embryo development restricted or non-existent. Such restricted growth and embryo abortion is found to be due to cessation of early endosperm development in A. duranensis (2x) times A. paraguariensis (2x), the non-development of endosperm beyond the coenocytic stage in A. batizocoi (2x) times A. paraguariensis (2x), and the overgrowth of nucellar tissue into the embryo sac in case of A. hypogaea (4x) times A. paraguariensis (2x).The weak cross-compatibility between the species of two sections suggest relatively closer phylogenetic relationship between them, than with the other incompatible sections of the genus Arachis.     

Discrimination of three Pegaga (Centella) varieties and determination of growth-lighting effects on metabolites content based on the chemometry of 1H nuclear magnetic resonance spectroscopy

The metabolites of three species of Apiaceae, also known as Pegaga, were analyzed utilizing (1)H NMR spectroscopy and multivariate data analysis. Principal component analysis (PCA) and hierarchical cluster analysis (HCA) resolved the species, Centella asiatica, Hydrocotyle bonariensis, and Hydrocotyle sibthorpioides, into three clusters. The saponins, asiaticoside and madecassoside, along with chlorogenic acids were the metabolites that contributed most to the separation. Furthermore, the effects of growth-lighting condition to metabolite contents were also investigated. The extracts of C. asiatica grown in full-day light exposure exhibited a stronger radical scavenging activity and contained more triterpenes (asiaticoside and madecassoside), flavonoids, and chlorogenic acids as compared to plants grown in 50% shade. This study established the potential of using a combination of (1)H NMR spectroscopy and multivariate data analyses in differentiating three closely related species and the effects of growth lighting, based on their metabolite contents and identification of the markers contributing to their differences.     

Novel flavonol glycoside, 7-O-methyl mearnsitrin, from Sageretia theezans and its antioxidant effect

A novel flavonol glycoside, 7-O-methylmearnsitrin (7,4'-O-dimethylmyricetin 3-O-alpha-L-rhamnopyranoside), and myricetrin, kaempferol 3-O-alpha-L-rhamnopyranoside, europetin 3-O-alpha-L-rhamnoside, and 7-O-methyl quercetin 3-O-alpha-L-rhamnopyranoside were isolated from the leaves of Sageretia theezans, and their chemical structures were identified by spectroscopic analyses including two-dimensional NMR (HSQC, HMBC). Whereas myricetrin, europetin 3-O-alpha-L-rhamnoside, and 7-O-methylquercetin 3-O-alpha-L-rhamnopyranoside showed stronger activities than ascorbic acid and alpha-tocopherol, 7-O-methylmearnsitrin showed very weak antioxidant activities by ESR and LDL oxidation inhibition tests.     

1H NMR based metabolic profiling in the evaluation of Japanese green tea quality

Classification of tea quality is now mainly performed according to the sensory results by professional tea tasters. However, this evaluation method is inconsistent in differentiating their qualities. A combination of a (1)H NMR technique and a multivariate analysis was introduced to the quality evaluation of green tea by means of a metabolomic technique. A broad range of metabolites were detected by (1)H NMR spectrometry. The principal component analysis (PCA) was used to reduce the complexity of the (1)H NMR spectra data set and provided the quality discrimination result. It offered an extensive clue for classification and quality assessment without any prepurification method. A set of green teas from a Japanese tea contest were analyzed by (1)H NMR to classify the quality with respect to that judged by tea tasters and to conceive a quality prediction model. Metabolic profiling and fingerprinting of (1)H NMR spectra of green teas with different quality were studied. PCA showed a separation between the high- and the low-quality green teas. The taste marker compounds contributing to the discrimination of tea quality were identified. Reliable prediction models were obtained by the partial least-squares projection to latent structure (PLS) analysis together with a preprocessing filter of both orthogonal signal correction (OSC) and a combination between OSC and wavelet transform algorithms.     

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.     

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.     

Metabolic characterization of Brassica rapa leaves by NMR spectroscopy

The Brassica has been intensively studied due to the nutritional and beneficial effects. However, many species, varieties, and cultivars of this genus and the resulting large metabolic variation have been obstacles for systematic research of the plant. In order to overcome the problems posed by the biological variation, the metabolomic analysis of various cultivars of Brassica rapa was performed by NMR spectroscopy combined with multivariate data analysis. Discriminating metabolites in different cultivars and development stages were elucidated by diverse 2D-NMR techniques after sorting out different significant signals using (1)H NMR measurements and principal component analysis. Among the elucidated metabolites, several organic and amino acids, carbohydrates, adenine, indole acetic acid (IAA), phenylpropanoids, flavonoids, and glucosinolates were found to be the metabolites contributing to the differentiation between cultivars and age of Brassica rapa. On the basis of these results, the distribution of plant metabolites among different cultivars and development stages of B. rapa is discussed.     

Larvicidal activity of lignans identified in Phryma leptostachya Var. asiatica roots against three mosquito species

The insecticidal activity of phytochemicals isolated from the roots of Phryma leptostachya var. asiatica against third instar larvae of Culex pipiens pallens, Aedes aegypti, and Ocheratatos togoi was examined. The two constituents of P. leptostachya var. asiatica roots were identified as the leptostachyol acetate (I) and 8'-acetoxy-2,2',6-trimethoxy-3,4,4',5'-dimethylenedioxyphenyl-7,7'-dioxabicyclo[3.3.0]octane (II) by spectroscopic analysis. Compound I was lethal to C. pipiens pallens, A. aegypti, and O. togoi at 10 ppm. Compound II showed weak or no insecticidal activity against three mosquito species at 10 ppm. The LC(50) values of I against C. pipiens pallens, A. aegypti, and O. togoi were 0.41, 2.1, and 2.3 ppm, respectively. Naturally occurring P. leptostachya var. asiatica root-derived compounds merit further study as potential mosquito larval control agents or lead compounds.     

Polymorphisms Among Chloroplast and Mitochondrial Genomes of <Emphasis Type="Italic">Citrullus</Emphasis> Species and Subspecies

Twelve and six DNA clones representing various parts of chloroplast and mitochondrial genomes, respectively, were used to detect polymorphism among five watermelon cultivars and 21 U.S. Plant Introductions (PIs) collected from diverse geographical locations and representing major groups of Citrullus species. Cluster analysis based on 20 chloroplast DNA (cpDNA) and 10 mitochondrial DNA (mtDNA) restriction fragment length polymorphism (RFLP) markers differentiated the accessions into three major phenetic groups: PIs and watermelon cultivars of Citrullus lanatus subsp. vulgaris (Schrad. ex Eckl. et Zeyh.) Fursa (also designated as C. lanatus var. lanatus) (group I), PIs of C. lanatus var. citroides (of C. lanatus subsp. lanatus Schrad. ex Eckl. et Zeyh.)(group II), and C. colocynthis (L.) Schrad. PIs (group III). The chloroplast and mitochondrial genomes of watermelon cultivars are distinct, but closely related to those of the C. lanatus var. lanatus PIs. On the other hand, the chloroplast and mitochondrial genomes of the wild species C. colocynthis are more similar to those of C. lanatus var. citroides. Polymorphic cpDNA and mtDNA markers identified in this study can complement isozyme and nuclear DNA data used in earlier phylogenetic and phenetic classifications of Citrullus PIs. These cpDNA and mtDNA markers are being used in experiments designed to enhance watermelon cultivars by replacing the chloroplast and mitochondrial genome of cultivated watermelon with those of the wild species C. colocynthis.     

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.     

Analyses of arbutin and chlorogenic acid, the major phenolic constituents in Oriental pear

The HPLC retention time, photodiode array UV spectrum analysis, and LC/MS results indicated that arbutin and chlorogenic acid are the main phenolic constituents in Oriental pear. The two compounds exist in different organs of the Yali pear, which is one of the major cultivars of Pyrus bretschnrideri. The contents of arbutin in the leaf bud, floral bud, flower, and young fruit were 11.9, 12.4, 8.29, and 9.92 mg/g fresh weight (FW), respectively. Chlorogenic acid amounts in the same organs were 2.26, 3.22, 5.32, and 3.72 mg/g FW, respectively. During development, the concentration of the two compounds in Yali pears was the greatest in young fruit (9.92 mg/g FW of arbutin and 3.72 mg/g FW of chlorogenic acid), and then declined swiftly with fruit growth to less than 0.400 and 0.226 mg/g FW, respectively, in mature fruit. Large differences existed in the distribution of the two compounds in parts of the mature fruit of 14 Oriental pear cultivars. The greatest concentration of arbutin was found in the peel (1.20 mg/g FW), which was 3-5 times greater than that found in the core and 10-45 times greater than the level in the pulp. The concentration of chlorogenic acid in the core was greater than that in the peel. The compounds in 17 cultivars of Oriental pear, including P. bretschnrideri, Pyrus pyrifolia, Pyrus ussuriensis, and Pyrus sinkiangensis, were compared with those in 5 cultivars of Occidental pear (Pyrus communis). The mean concentration of arbutin in the Oriental pear cultivars was 0.164 mg/g FW, greater than the 0.083 mg/g FW found in the Occidental pear cultivars. The greatest arbutin content was 0.400 mg/g FW, found in the Yali pear. However, the mean concentration of chlorogenic acid in the Oriental pear was 0.163 mg/g FW, less than that found in the Occidental pear (0.309 mg/g FW).     

不同地理种群巴山榧树及近缘种叶绿体trnL-trnF序列的分析

本文运用PCR产物直接测序法对巴山榧树10个地理种群的trnL-trnF序列进行了测定,并调用GenBank中6个近缘种的trnL-trnF序列,选用MEGA 4.1软件对巴山榧树不同地理种群及近缘种间的trnL-trnF序列进行分析。结果表明,巴山榧树10个地理种群间的遗传分化程度较低,只有34个变异位点,1个信息位点;7个近缘种间的遗传分化较显著,有75个变异位点,30个信息位点。聚类分析将巴山榧树10个地理种群与6个近缘种聚为2个分支,巴山榧树、云南榧和日本榧树聚为一支,榧树、长叶榧树、佛罗里达榧和加州榧聚为另一个分支。trnL-trnF序列系统树不支持将榧树属分为皱乳榧组和榧树组;赞同将云南榧并入巴山榧树,或作为巴山榧树的变种处理;不支持将巴山榧树和云南榧合并在榧树下作为变种的处理意见。     

Composition of phenolic compounds in a Portuguese pear (Pyrus communis L. var. S. Bartolomeu) and changes after sun-drying

The composition of phenolic compounds of a Portuguese pear cultivar (Pyrus communis L. var. S. Bartolomeu) was determined by HPLC after thioacidolysis. The average concentration of phenolic compounds in pear harvested at commercial maturity stage was 3.7 g per kg of fresh pulp. Procyanidins were the predominant phenolics (96%), with a mean degree of polymerization (mDP) of 13-44; hydroxycinnamic acids (2%), arbutin (0.8%), and catechins (0.7%) were also present. The most abundant monomer in the procyanidin structures was (-)-epicatechin (99%), which was found as extension and terminal units; (+)-catechin (1%) was found only as a terminal unit. Sun-drying of these pears caused a decrease of 64% (on a dry pulp basis) in the total amount of native phenolic compounds. Hydroxycinnamic acids and procyanidins showed the largest decrease; the B2 procyanidin was not found at all in the sun-dried pear. Less affected were arbutin and catechins. In the sun-dried pear, the procyanidins with high mDP became unextractable in the solvents used.