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.     

Immunostimulating activity of a crude polysaccharide derived from green tea (Camellia sinensis) extract

Green tea extract is well-known to reduce the risk of a variety of diseases. Here, we investigated the immunostimulating activity of tea polysaccharide (TPS), one of the main components in green tea extract. The water extracts from mature or immature tea leaves were precipitated by using ethanol at room temperature. The sediment was washed with ethanol and acetone alternately and then dried. We used the phagocytic activity of macrophage-like cells as an indicator of immune function activation. Chemical components were analyzed by HPLC. The immunostimulating activity of TPS from immature leaves extract was higher than that of TPS from mature leaves, and its activities were dependent on the content of strictinin in the leaf extract. Futhermore, a mixture of catechin and TPS that removed polyphenols did not increase the immunostimulating activity. These results suggest that the catechin-polysaccharide complex is a very important molecule in the immunomodulating activity of tea extracts.     

The impact of packaging materials on the antioxidant phytochemical stability of aqueous infusions of green tea (Camellia sinensis) and yaupon holly (Ilex vomitoria) during cold storage

Ready to drink (RTD) teas are a growing segment in the beverage category, brought about by improvements in the flavor of these products and healthy market trends driven by consumers. The presented results evaluated the antioxidant phytochemical stability of RTD teas from aqueous infusions of traditional green tea (Camellia sinensis) and a botanical tea from yaupon holly (Ilex vomitoria) as influenced by packaging materials during cold storage. Two common packaging materials for RTD products are glass and polyethylene terephthalate (PET) and have been compared to a retortable pouch (RP), an emerging packaging material for various types of food since it is durable, inexpensive, lightweight, and easy to sterilize. Storage stability was then evaluated for each aqueous infusion prepared at 10 g/L at 90 °C for 10 min and evaluated at 3 °C in the absence of light over 12 weeks. Analyses included quantification and characterization of individual polyphenolics by high-performance liquid chromatography-photodiode array and liquid chromatography-electrospray ionization-mass spectrometry as well as changes in total antioxidant capacity. For green tea, concentrations of the three major flavan-3-ols, epigallocatechin gallate, epigallocatechin, and epicatechin gallate were better retained in glass bottles as compared to other packages over 12 weeks. In yaupon holly, chlorogenic acid and its isomers that were the predominant compounds were generally stable in each packaging material, and a 20.6-fold higher amount of saponin was found as compared to green tea, which caused higher stability of flavonol glycosides present in yaupon holly during storage. The antioxidant capacity of green tea was better retained in glass and PET versus RP, whereas no differences were again observed for yaupon holly. Results highlight the superiority of oxygen-impervious glass packaging, but viable alternatives may be utilizable for RTD teas with variable phytochemical compositions.     

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.     

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.     

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.     

Total polyphenol content and antioxidant capacity of commercially available tea (Camellia sinensis) in Argentina

Tea, Camellia sinensis (L.) O. Kuntze (Theaceae) is cultivated in Argentina in the northeastern region (provinces of Misiones and Corrientes), between 26 degrees and 28 degrees south latitude, the southernmost area of the world where tea is cultivated. The objective of this work was to determine the total polyphenol content and the in vitro antioxidant capacity of green and black tea cultivated and industrialized in Argentina. Twelve samples of eight brands were analyzed. The total polyphenol content was determined according to the International Organization for Standardization method (ISO) 14502-1 for the determination of substances characteristic of green and black tea. The antioxidant capacity was determined by the ferric thiocyanate method (FTC) and the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free-radical scavenging assay. Green tea showed a higher polyphenol content than black tea. The total polyphenol concentration in green tea was found to vary from 21.02 +/- 1.54 to 14.32 +/- 0.45% of gallic acid equivalents (GAE), whereas in black tea, the polyphenol content ranged from 17.62 +/- 0.42 to 8.42 +/- 0.55% of GAE (P < 0.05). A similar profile was observed for the antioxidant capacity determined by both methods. The antioxidant activities were well correlated with the total polyphenol content (r (2) = 0.9935 for the ferric thiocyanate method and r (2) = 0.9141 for the 1,1-diphenyl-2-picrylhydrazyl free-radical scavenging assay). This is the first systematic screening for the quantification of polyphenols and antioxidant activity in tea commercialized in Argentine markets. The results obtained herein allow one to conclude that Argentine tea is of very good quality when compared to teas from other sources.     

Characterization of the constituents and antioxidant activity of Brazilian green tea (Camellia sinensis var. assamica IAC-259 cultivar) extracts

Freeze-dried extracts from Camellia sinensis var. assamica IAC-259 cultivar named Brazilian green tea were prepared by hot water and ultrasound-assisted extractions using leaves harvested in spring and summer. Their caffeine and catechin contents were measured by high performance liquid chromatography-diode array detector. The antioxidant activity of the major green tea compounds and Brazilian green tea extracts was evaluated using a 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The levels of caffeine were higher in the summer samples (p < 0.05); otherwise, there were no significant variations related to the catechin contents between spring and summer samples. The sonication method using water/acetone as solvent had a high efficiency to extract not only epigallocatechin gallate but also epicatechin gallate (p < 0.05). Antioxidant activities of the Brazilian green tea extracts were not significantly different among seasons and extraction systems. The antioxidant data (IC50) of the Brazilian green tea extracts showed a significant correlation with their epigallocatechin gallate and epicatechin gallate contents (p < 0.05).     

Modulation of hepatic drug metabolizing enzymes and oxidative status by rooibos (Aspalathus linearis) and Honeybush (Cyclopia intermedia), green and black (Camellia sinensis) teas in rats

Rooibos and honeybush teas significantly (P < 0.05) enhanced the activity of cytosolic glutathione S-transferase alpha. A significant (P < 0.05) to marginal (P < 0.1) increase in the activity of the microsomal UDP-glucuronosyl transferase was obtained with unprocessed rooibos and honeybush teas, respectively. Oxidized glutathione (GSSG) levels were significantly (P < 0.05) reduced in the liver of all tea treated rats while reduced glutathione (GSH) was markedly increased in the liver of the herbal tea treated rats. These changes resulted in a significant (P < 0.05) increase in the GSH/GSSG ratio by the unprocessed, processed rooibos and unprocessed honeybush teas. Green and black teas markedly to significantly decreased the oxygen radical absorbance capacity in liver homogenates, respectively. Modulation of phase II drug metabolizing enzymes and oxidative status in the liver may be important events in the protection against adverse effects related to mutagenesis and oxidative damage.     

Characterization of phenolic compounds in strawberry (Fragaria x ananassa) fruits by different HPLC detectors and contribution of individual compounds to total antioxidant capacity

Phenolic compounds in strawberry (Fragaria x ananassa) fruits were identified and characterized by using the complementary information from different high-performance liquid chromatography detectors: diode array, mass spectrometer in positive and negative mode, and coulometric array. Electrochemical profiles obtained from the coulometric array detector contributed to the structural elucidation suggested from the UV-vis and mass spectra. About 40 phenolic compounds including glycosides of quercetin, kaempferol, cyanidin, pelargonidin, and ellagic acid, together with flavanols, derivatives of p-coumaric acid, and ellagitannins, were described, providing a more complete identification of phenolic compounds in strawberry fruits. Quercetin-3-malonylhexoside and a deoxyhexoside of ellagic acid were reported for the first time. Antioxidative properties of individual components in strawberries were estimated by their electrochemical responses. Ascorbic acid was the single most important contributor to electrochemical response in strawberries (24%), whereas the ellagitannins and the anthocyanins were the groups of polyphenols with the highest contributions, 19 and 13% at 400 mV, respectively.     

Modified 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (abts) method to measure antioxidant capacity of Selected small fruits and comparison to ferric reducing antioxidant power (FRAP) and 2,2'-diphenyl-1-picrylhydrazyl (DPPH) methods

The measurement of antioxidant capacity in fruits differs from that of other biological samples due to their low pH and very low lipophilic antioxidant capacity. In this report, we present a modified 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) method for fruits and compare its performance with the other commonly used antioxidant methods of 2,2'-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP). The antioxidant capacity and reaction kinetics of four phenolic compounds, two antioxidant standards, and five fruits were also investigated. The modified ABTS method prepared at a pH of 4.5 with sodium acetate buffer is highly stable and easily applied to fruit samples as compared to the standard (pH 7.4) version. The measured antioxidant capacity of samples varied with the assay method used, pH, and time of reaction. Traditional antioxidant standards (trolox, ascorbic acid) displayed stable, simple reaction kinetics, which allowed end point analysis with all of assays. Of the phenolic compounds examined, chlorogenic and caffeic acids exhibited the most complex reaction kinetics and reaction rates that precluded end point analysis while gallic acid and quercetin reached stable end points. All fruit extracts exhibited complex and varied kinetics and required long reaction times to approach an end point. Because the antioxidant capacity of fruit extracts is a function of the array of individual antioxidants present, accurate comparisons among fruit samples require that reaction times be standardized and of sufficient length to reach steady state conditions and that more than one assay be used to describe the total antioxidant activity of fruit samples.     

Measuring antioxidant efficiency of wort, malt, and hops against the 2,2'-azobis(2-amidinopropane) dihydrochloride-induced oxidation of an aqueous dispersion of linoleic acid

This paper presents a simple, convenient method for determining the efficiency of antioxidants in aqueous systems. Production of conjugated diene hydroperoxide by oxidation of linoleic acid in an aqueous dispersion is monitored at 234 nm. 2, 2'-Azobis(2-amidinopropane) dihydrochloride is used as a free radical initiator. Among 12 antioxidants tested, phenolic compounds proved to be the most efficient, both kinetically and in terms of the inhibition time (T(inh)). Applied to wort, malt, and hops, the method confirmed a significant antioxidant activity in such products, especially hops. This assay can be used to follow oxidative changes throughout the brewing process and to understand the contribution of each raw material.     

Application of artificial neural network (ANN) and partial least-squares regression (PLSR) to predict the changes of anthocyanins, ascorbic acid, Total phenols, flavonoids, and antioxidant activity during storage of red bayberry juice based on fractal analysis and red, green, and blue (RGB) intensity values

Artificial neural network (ANN) and partial least-squares regression (PLSR) models were developed to predict the changes of anthocyanin (AC), ascorbic acid (AA), total phenols (TP), total flavonoid (TF), and DPPH radical scavenging activity (SA) in bayberry juice during storage based on fractal analysis (FA) and red, green, and blue (RGB) intensity values. The results show the root mean squared error (RMSE) of ANN-FA decreased 2.44 and 12.45% for AC (RMSE = 18.673 mg/100 mL, R(2) = 0.939) and AA (RMSE = 8.694 mg/100 mL, R(2) = 0.935) compared with PLSR-RGB, respectively. In addition, PLSR-FA (RMSE = 5.966%, R(2) = 0.958) showed a 12.01% decrease in the RMSE compared with PLSR-RGB for predicting SA. For the prediction of TP and TF, however, both models showed poor performances based on FA and RGB. Therefore, ANN and PLSR combined with FA may be a potential method for quality evaluation of bayberry juice during processing, storage, and distribution, but the selection of the most adequate model is of great importance to predict different nutritional components.     

Evaluation of ecological doses of the nitrification inhibitors 3,4-dimethylpyrazole phosphate (DMPP) and 4-chloromethylpyrazole (ClMP) in comparison to dicyandiamide (DCD) in their effects on dehydrogenase and dimethyl sulfoxide reductase activity in soils

Risk assessment of the nitrification inhibitors (NIs) 3,4-dimethylpyrazole phosphate (DMPP), 4-chloromethylpyrazole (ClMP), and dicyandiamide (DCD) on nontarget microbial activity in soils was determined by measuring dehydrogenase and dimethyl sulfoxide reductase activity (DHA, DRA, respectively) in three differently textured soils under laboratory conditions. Dehydrogenase activity was measured with standard procedure recommended to evaluate side effects of environmental chemicals on general microbial activity in soils. The kinetic of inhibition were obtained by dose–response relationships and used to calculate the no observable effect levels (NOEL values) and the effective doses at 10% and 50% inhibition (ED₁₀ and ED₅₀), respectively. Negative effects on DHA and DRA, respectively, were observed only at rates approximately 40–100 times higher than the concentrations recommended in the field. Both DHA and DRA were affected more in the sandy than in the silty or clayey soil. Consequently, NOEL, ED₁₀, and ED₅₀ values were considerably higher in the clayey than in the silty or sandy soil. The heterocyclic N compounds DMPP and ClMP, respectively, were more effective in inhibiting DHA and DRA than DCD. At application rates used in the field as well as at concentration up to 25 to 90 times higher, the NIs concerned failed to affect general soil microbial activity in soils. Among the three NIs tested, the not marketed ClMP exhibited the strongest negative effects on soil microbial activity. At recommended application rates, the NIs tested should be considered as enviromentally safe.     

Application of the 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation assay to a flow injection system for the evaluation of antioxidant activity of some pure compounds and beverages

The 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation (ABTS(*)(+)) assay was adapted to a flow injection (FI) system to obtain a sensitive and rapid technique for the monitoring of antioxidant activity of pure compounds and complex matrixes, such as beverages and food extracts. The FI system includes a HPLC pump that flows the mobile phase (a solution of ABTS(*)(+) in ethanol) through a 20 microL loop injector, a single bead string reactor filled with acid-washed silanized beads, a delay coil and a photodiode array UV-visible detector. The technique was very sensitive, with limits of detection and of quantification of 4.14 and 9.29 micromol of Trolox/L, respectively, and demonstrated high repeatability and reproducibility. The proposed technique was then applied to the evaluation of the antioxidant activity of some pure compounds, demonstrating good agreement with published data obtained by the original spectrophotometric ABTS(*)(+) assay. Finally, the total antioxidant activity of 10 beverages was determined by both the proposed and the original method. The values ranged from 0.09 mmol L(-)(1) for cola to 49.24 mmol L(-)(1) for espresso coffee and did not result significantly different from those obtained by the original spectrophotometric ABTS(*)(+) assay (Student's paired t-test: t = 1.4074, p = 0.1929). In conclusion, the proposed FI technique seems suitable for the direct, rapid and reliable monitoring of total antioxidant activity of pure compounds and beverages and, due to the ability to operate in continuous, it allows the analysis of about 30 samples h(-)(1) making the assay particularly suitable for large screening of total antioxidant activity in food samples.