MODEL DEVELOPMENT AND PROCESS OPTIMIZATION FOR SOLVENT EXTRACTION OF POLYPHENOLS FROM RED GRAPES USING BOX–BEHNKEN DESIGN

The objective of the present study is to find out the optimum extraction conditions for extraction of polyphenols from red grapes using Box–Behnken design. Red grapes polyphenols were extracted using acid–ethanol solvent at various extraction temperature (40–60°C), extraction time (20–100 min) and different solid–liquid ratio (1:5–1:15 g:ml). The effect (main and interactive) of extraction conditions on total anthocyanin, phenolic and flavonoid content were studied using Box–Behnken design (three factors at three levels). The results showed that the contribution of the quadratic model was significant for all the responses. Second-order mathematical regression models were developed and were found to fit well with observed data. Derringer's desirability function methodology was performed to find out the optimal conditions based on both individual and combinations of all responses (extraction temperature: 57°C, time: 61 min, and solid–liquid ratio: 1:8.7 g:ml) were established. At this optimal condition, the anthocyanin yield, total phenolic and flavonoid content were 73.92 mg/100 g, 221.4 mg GAE/100 g, and 79.08 mg CE/100 g, respectively. A desirability value of 0.902 was achieved at this point.     

Bioreactor type affects the accumulation of phenolic acids and flavonoids in microshoot cultures of Schisandra chinensis (Turcz.) Baill.

Microshoots of the East Asian medicinal plant species Schisandra chinensis (Chinese magnolia vine) were grown in bioreactors characterized by different construction and cultivation mode. The tested systems included two continuous immersion systems—a cone-type bioreactor (CNB) and a cylindric tube bioreactor (CTB), a nutrient sprinkle bioreactor (NSB), and two temporary immersion systems (TIS)—RITA® and Plantform. Microshoots were grown for 30 and 60 days in the MS medium enriched with 1 mg l^?1 NAA and 3 mg l^?1 BA. The accumulation of two groups of phenolic compounds: phenolic acids and flavonoids in the bioreactor-grown S. chinensis biomass, was evaluated for the first time. In the microshoot extracts, seven phenolic acids: chlorogenic, gallic, p–hydroxybenzoic, protocatechuic, syringic, salicylic and vanillic, and three flavonoids: kaempferol, quercitrin and rutoside, were identified. The highest total amount of phenolic acids (46.68 mg 100 g^?1 DW) was recorded in the biomass maintained in the CNB for 30 days. The highest total content of flavonoids (29.02 mg 100 g^?1 DW) was found in the microshoots maintained in the NSB for 30 days. The predominant metabolites in all the tested systems were: gallic acid (up to 10.01 mg 100 g^?1 DW), protocatechuic acid (maximal concentration 16.30 mg 100 g^?1 DW), and quercitrin (highest content 21.00 mg 100 g^?1 DW).

     

Optimisation of phenolic compounds and antioxidant activity extraction conditions of a roasted mix of Tetrapleura tetraptera (Schumach & Thonn.) and Aframomum citratum (C. Pereira) fruits using response surface methodology (RSM)

The therapeutic abilities of Tetrapleura tetraptera and Aframomum citratum fruits used as spices are attributed to their bioactive molecules, including polyphenols. Sometimes used together and heated, they can undergo denaturation. The aim of the current study is to optimize the extraction of phenolic compounds and antioxidant potential of a roasted mix of Tetrapleura tetraptera and Aframomum citratum (95/5: w/w) fruits using RSM in a home food consumption context. The mix of spices was chosen according to the highest content of TPP and preliminary studies were performed to select the influencing variables. Roasting temperatures (130–170 °C), roasting times (10–15 min) and brewing times (8–15 min) were investigated with a rotatable central composite design. Experimental results were fitted to the second-order polynomial model where multiple regressions and ANOVA were used to determine the coefficients of the model and the optimal conditions for the considered responses. The two spices are good sources of phenolic compounds, and they also show significant (p < 0.05) dose-dependent radical scavenging activities (DPPH assay and inhibition of β-carotene discoloration) and reductive activities (FRAP assay and Phosphomolybdenum method). They significantly inhibit bovine serum albumin and 5-LOX denaturation. Brewing time and roasting time significantly (p < 0.05) influence the responses and there is a strong (R² = 0.93) correlation between the TPP and TAC of the beverage. The quadratic model fit well and the different factors used to test its accuracy and fitness were in satisfactory ranges. For TPP extraction (38.90 mgGAE/g dw) and TAC (50.75 mg TE/g dw) expression, the optimal conditions were reached at a roasting temperature of 150 °C, roasting time of 12.62 min, brewing time of 11.91 min and a desirability of 0.95. The novel information on the optimisation of the process can be further used by scientists, consumers and herbalists for effective handling of fruits during the extraction process.     

Modeling of polyphenols extraction from pomegranate by-product using rotatable central composite design of experiments

There is a growing request to find an effective method of polyphenols extraction from agro-industry by-product as pomegranate. In this study, response surface methodology (RSM) was used to explore the effect of three factors on ultrasonic assisted extraction (UAE) of total polyphenols (TP), total flavonoids (TF) and condensed tannins (CT) from pomegranate peels. The optimal conditions were determined for each phenolic compound using regression model equations and 3-D plots. The high TP, TF and CT content were obtained with, respectively, liquid/solid ratio of 20.00, 9.77, 9.77, extraction time of 36.38, 41.82, 30.39 min and 36.00, 83.64, 59.26% of ethanol percentage. In fact, liquid/solid ratio of 20, extraction time of 30.94 min and 59.26% of ethanol gives the highest contents of TP, TF and CT simultaneously. The experimental values using optimal conditions agreed with the predicted values. The pomegranate peels extract obtained under optimum conditions have an effective antioxidant activity as determined by ABTS and DPPH assays.     

Phenolic Profile of Artemisia alba Turra

The aim of this study was to evaluate flower and leaf methanol extracts of Artemisia alba Turra for their total phenolic and flavonoid contents, antioxidant capacity and to investigate their phenolic composition. The flower extract was richer in total phenolics and flavonoids and possessed higher antioxidant activity through DPPH and ABTS assays. The UHPLC‐PDA‐MS analysis of the flower and leaf methanol extracts revealed similar phenolic profile and allowed identification of 31 phenolic compounds (flavonoids, coumarins, and phenolic acids) by comparison with the respective reference compounds or tentatively characterized by their chromatographic behavior, UV patterns, and MS fragmentations. The presence of hispidulin, jaceosidin, desmethoxycentaureidin, and dicaffeoyl esters of quinic acid in A. alba is reported herein for the first time. The distribution of flavonoids in A. alba from different origins was discussed from chemotaxonomic point of view.     

Optimization of extraction method for LC–MS based determination of phenolic acid profiles in different Impatiens species

The main aim of presented study was the comparison of various extraction methods for the quantitative and qualitative analysis (LC-ESI–MS/MS) of phenolic acids present in extracts obtained from leaves, flowers, and roots of Impatiens glandulifera. The accelerated solvent extraction (ASE) at three temperature ranges (80° C, 100° C, and 120° C), ultrasound assisted extraction (USAE) at 60° C, and traditional extraction in Soxhlet apparatus were used. Taking into account the extraction yield, and the diversity of the individual compounds, ultrasound assisted extraction proved to be the most efficient method, and it was used to determine the content of phenolic acids in leaves of four other Impatiens species, including I. balsamina, I. noli-tangere, I. parviflora, and I. walleriana. Eleven phenolic acids were identified in all examined species. These were protocatechuic, gentisic, 4- hydroxybenzoic, vanillic, trans-caffeic, syringic, trans-p-coumaric, trans- and cis-ferulic, salicylic, and 3-hydroxycinnamic acids. In the extract from the leaves of I. balsamina and I. walleriana, gallic and cis-p-coumaric acids were found additionally. The most abundant compounds in all examined extracts were protocatechuic and 3-hydroxycinnamic acids. The latest acid was found in the highest yield in I. noli-tangere (266.12 μg/g DW). In the leaves of I. glandulifera a great amount of 4-hydroxybenzoic (41.44 μg/g DW), vanillic (61.50 μg/g DW), and trans-p-coumaric (58.42 μg/g DW) acids was also observed. Our results indicate that protocatechuic, 4-hydroxybenzoic, vanillic, trans-p-coumaric, trans-ferulic, and 3-hydroxycinnamic acids were most characteristic of Impatiens species.Additionally, various phenolic-rich extracts from leaves, flowers, and roots of Impatiens glandulifera were tested for antioxidant activity. The highest antiradical activity was detected for roots using Soxhlet extraction (EC50 = 0.055 mg [DE/ml]).The study demonstrated that members of the genus Impatiens, and in particular Impatiens glandulifera, and Impatiens noli-tangere, contain significant amounts of phenolic acids. In addition, extracts from various parts of I. glandulifera could be interesting as novel sources of natural antioxidants.     

Physical and chemical properties of gelatin from the skin of cultured Amur sturgeon (Acipenser schrenckii)

To expand the usefulness of cultured Amur sturgeon, Acipenser schrenckii, its skin was used to explore the production of gelatin. After acetic acid pre‐treatment (0.05 m for 3 h), gelatin was extracted at temperatures of 50 or 70°C for 1 or 6 h. Gelatin yield ranged from 9.42 to 12.47% (wet weight basis) (P < 0.05). With increasing extraction time and temperature, the content of imino acids (proline + hydroxyproline), gel strength and L*‐value (lightness) decreased, while the a*‐value (redness) and b*‐value (yellowness) of gelatin gel increased (P < 0.05). Electrophoretic analysis revealed that α‐chains and β‐chains were predominant components in all extracted gelatins. Higher molecular weight proteins (γ‐chain) were also observed. Gelling and melting temperatures of gelatin were 13.6–14.6°C and 20.3–22.6°C, respectively. Circular dichroism (CD) spectra and Fourier transform infrared (FTIR) spectroscopy revealed the triple helix loss in gelatin (A₁₂₃₅ (AIII)/A₁₄₅₁ < 1). Extraction conditions caused secondary structure changes in the gelatin. More likely due to the differences in the culture water temperature, gelatin exhibited gelling and melting temperatures intermediate between those of cold‐ and warm‐water fish gelatins. The obtained gelatin can be used in food products or in the production of bioactive compounds.     

Optimization of ultrasound-assisted extraction of pectinase enzyme from guava (Psidium guajava) peel: Enzyme recovery,specific activity,temperature, and storage stability

This study aimed to investigate the effects of the ultrasound-assisted extraction conditions on the yield, specific activity, temperature, and storage stability of the pectinase enzyme from guava peel. The ultrasound variables studied were sonication time (10–30 min), ultrasound temperature (30–50°C), pH (2.0–8.0), and solvent-to-sample ratio (2:1 mL/g to 6:1 mL/g). The main goal was to optimize the ultrasound-assisted extraction conditions to maximize the recovery of pectinase from guava peel with the most desirable enzyme-specific activity and stability. Under the optimum conditions, a high yield (96.2%), good specific activity (18.2 U/mg), temperature stability (88.3%), and storage stability (90.3%) of the extracted enzyme were achieved. The optimal conditions were 20 min sonication time, 40°C temperature, at pH 5.0, using a 4:1 mL/g solvent-to-sample ratio. The study demonstrated that optimization of ultrasound-assisted process conditions for the enzyme extraction could improve the enzymatic characteristics and yield of the enzyme.     

Phytogenic compounds from avocado (Persea americana L.) extracts; antioxidant activity,amylase inhibitory activity,therapeutic potential of type 2 diabetes

Diabetes is a worldwide public health disease. Currently, the most effective way to treat diabetes is to mitigate postprandial hyperglycemia by inhibiting carbohydrate hydrolysis enzymes in the digestive system. Plant extracts are rich in bioactive compounds, which can be used in diabetes treatment. This study aims to evaluate the polyphenols content in ethanolic extracts of avocado fruit and leaves (Persea americana Mill.). Additionally, their antioxidant activity using DPPH, while the inhibition ability of α-amylase was examined by reacting different amounts of the extracts with α-amylase compared to acarbose as standard inhibitor. The active compounds were detected in the extracts by LC/MS. The obtained results showed that the leaf extract recorded a significant content of total phenolic compounds compared to the fruit extract (178.95 and 145.7 mg GAE /g dry weight, respectively). The total flavonoid values ??ranged from 32.5 to 70.08 mg QE/g dry weight of fruit and leaves extracts, respectively. Twenty-six phytogenic compounds were detected in leaf and fruit extract by LC/MS. These compounds belong to fatty acids, sterols, triterpenes, phenolic acids, and flavonoids. The antioxidant activity of the extracts is due to the exist of phytogenic compounds, i.e., polyphenols and flavonoids. The antioxidant activity increased in a concentration dependant manner. Avocado fruit extract (1000 µg/mL) scavenged 95% of DPP? while leaf extract rummaged 91.03% of free radicals compared with Vit C and BHT. Additionally, higher α-amylase inhibitory activity was observed in fruit extract than the leaf extract, where the fruit and leaf extract (1000 μg/ml) inhibited the enzyme by 92.13% and 88.95%, respectively. The obtained results showed that the ethanolic extracts of avocado could have a significant impact on human health due to their high content of polyphenols.     

Enzyme-assisted extraction of lycopene from tomato processing waste

A central composite design was used to optimize the enzyme-assisted extraction of lycopene from the peel fraction of tomato processing waste. Tomato skins were pretreated by a food-grade enzyme preparation with pectinolytic and cellulolytic activities and then subjected to hexane extraction. The factors investigated included extraction temperature (10–50 °C), pretreatment time (0.5–6.5 h), extraction time (0.5–4.5 h), enzyme solution-to-solid ratio (10–50 dm³/kg) and enzyme load (0–0.2 kg/kg). Overall, an 8- to 18-fold increase in lycopene recovery was observed compared to the untreated plant material. From a response surface analysis of the data, a second-degree polynomial equation was developed which provided the following optimal extraction conditions: T = 30 °C, extraction time = 3.18 h and enzyme load = 0.16 kg/kg. The obtained results strongly support the idea of using cell-wall degrading enzymes as an effective means for recovering lycopene from tomato waste.     

Development of a protocol for supercritical carbon dioxide extraction of ubiquinone-10 from dried biomass of <Emphasis Type="Italic">Pseudomonas diminuta</Emphasis>

Ubiquinone (Coenzyme Q10, CoQ10), a yellow-to-orange-colored lipophilic substance having nutraceutical value, was extracted from dried biomass of Pseudomonas diminuta using supercritical carbon dioxide (SC-CO₂). The effect of different operational parameters (temperature, pressure, and extraction time) and addition of co-solvent on SC-CO₂ extraction of CoQ10 was studied in detail. The solubility parameter of CoQ10, CO₂, and CO₂ with ethanol and methanol as co-solvents was calculated and validated with experimental results. Theoretically, ethanol and methanol had significant effect as co-solvent, and the difference between the two was only marginal. A maximum recovery of 22.33% was obtained using pure SC-CO₂ at 40 °C, 150 bar, and run time of 60 min. Ethanol as co-solvent at 3 mL/g of dried biomass at 40 °C and 150 bar increased the recovery from 22.33 to 68.57%. Further optimization of the extraction conditions by Box–Behnken design effectively increased the recovery to 96.2%. The optimized conditions were a temperature of 38 °C, pressure of 215 bar, and run time of 58 min.     

Microwave‐assisted water extraction of green tea polyphenols

Introduction – Green tea, a popular drink with beneficial health properties, is a rich source of specific flavanols (polyphenols). There is a special interest in the water extraction of green tea polyphenols since the composition of the corresponding extracts is expected to reflect the one of green tea infusions consumed worldwide. Objective – To develop a microwave‐assisted water extraction (MWE) of green tea polyphenols. Methodology – MWE of green tea polyphenols has been investigated as an alternative to water extraction under conventional heating (CWE). The experimental conditions were selected after consideration of both temperature and extraction time. The efficiency and selectivity of the process were determined in terms of extraction time, total phenolic content, chemical composition (HPLC‐MS analysis) and antioxidant activity of the extracts. Results – By MWE (80°C, 30 min), the flavanol content of the extract reached 97.46 (± 0.08) mg of catechin equivalent/g of green tea extract, vs. only 83.06 (± 0.08) by CWE (80°C, 45 min). In particular, the concentration of the most bioactive flavanol EGCG was 77.14 (± 0.26) mg of catechin equivalent/g of green tea extract obtained by MWE, vs 64.18 (± 0.26) mg/g by CWE. Conclusion – MWE appears more efficient than CWE at both 80 and 100°C, particularly for the extraction of flavanols and hydroxycinnamic acids. Although MWE at 100°C typically affords higher yields in total phenols, MWE at 80°C appears more convenient for the extraction of the green tea‐specific and chemically sensitive flavanols. Copyright © 2009 John Wiley & Sons, Ltd.     

A Spectrophotometric Analysis of Pigments in Apples

Methods of pigment extraction using traditional polar organic solvents (acetone or methanol) were compared to those employing a chloroform–methanol mixture. We found that, for spectrophotometric pigment analysis in the apple peel, the cuticular lipids must be preliminarily extracted from the samples with chloroform and MgO must be added during homogenization to prevent pigment degradation. The traditional extraction did not result in the complete extraction of intact pigments, and such extracts contained a considerable amount of light-absorbing impurities. The application of chloroform–methanol extraction allowed us to markedly reduce the content of such impurities and to increase the accuracy and sensitivity of the measurement of the content of chlorophylls and carotenoids. In addition, this extraction method proved useful for the analysis of phenolic substances (anthocyanins and flavonoids) in the water-methanol fraction of the extracts.     

Response Surface Methodology-Based Optimized Ultrasonic-Assisted Extraction and Characterization of Selected High-Value Components from Gemlik Olive Fruit

This article presents an optimized ultrasound-assisted ethanolic extraction (UAEE) and characterization of selected high-value components from Gemlik olive fruit (GOF) harvested from Potohar region of Pakistan. Response surface methodology (RSM), involving central composite design (CCD), was applied to optimize the extraction variables i. e., temperature (25–65 °C), extraction time (15–45 min) and aqueous ethanol concentration (60–90 %) for optimal recovery of bioactives extract, total phenolic contents (TPC) and DPPH free radical scavengers. Under the optimized set of conditions such as 43 °C temperature, 32 min extraction time and 80 % aqueous ethanol, the best extract yield (218.82 mg/g), TPC (19.87 mg GAE/g) and DPPH scavenging activity (63.04 %) were recorded. A quadratic polynomial model was found to be reasonably fitted to the observed results for extract yield (p<0.0001 and R²=0.9941), TPC (p<0.0001 and R²=0.9891), and DPPH radical scavenging activity (p<0.0001 and R²=0.9692). Potent phenolic compounds were identified by GC/MS in GOF extract and considerable amount of essential fatty acids were also detected. The current findings support the use of UAEE as an effective green route for optimized recovery of high-value components from GOF and hence its applications can be extended to functional food and nutra-pharmaceutical developments.     

Improved functional and nutritional properties of tomato fruit during cold storage

The use of synthetic antioxidants has been associated with serious concerns for human and environmental health. During ripening stages, tomato fruit is exposed to different abiotic stresses which not only influence its nutritional, mechanical, and functional properties at harvest, but also affect the quality and shelf life of the fruit during storage. This study investigated the pattern of changes in dietary antioxidants during various ripening stages of tomato fruit (cv. Red Rose) and their impact on storage behavior of the fruit during cold storage. Tomato fruits were harvested at mature green, breaker, turning, pink, light-red and red stages of maturity. Then, they were analysed for flesh firmness, soluble solids content, titratable acidity, total sugars, pH, dry matter content, lipophilic (lycopene, β-carotene, and total carotenoids), and hydrophilic (ascorbic acid, phenolic and flavonoids) antioxidants. Additional fruits were harvested at each maturity stage and divided into three equal lots, then were subjected to low-temperature (10 ± 1 °C) storage with 80 ± 5% RH, for 7, 14, and 21 days. Flesh firmness, and the levels of dietary antioxidants were analysed following the subsequent storage periods. The results revealed that the peak of hydrophilic antioxidants such as ascorbic acid, phenolic compounds, and flavonoids was between the ‘pink’ and the ‘light-red’ stages of fruit maturity. Whereas tomatoes harvested at the ‘red’ stage of maturity had the highest levels of lycopene and β-carotene. Both the stage of fruit maturity at harvest and duration of cold storage influenced flesh firmness, organoleptic and functional properties of ‘Red Rose’ tomato fruit. In conclusion, the results of the current investigation have practical implications in formulating foods with improved functional properties at processing industries.     

Comparison of thermotolerance of sun-exposed peel and shaded peel of ‘Fuji’ apple

The thermotolerance of the sun-exposed peel and the shaded peel of ‘Fuji’ apple (Malus domestica Borkh.) fruit was evaluated by measuring pigments, chlorophyll a fluorescence transients and O₂ evolution or uptake after exposure to 25, 35, 40, 42, 44, 46 or 48 °C for 30 min in the dark. A major effect of heat stress at 46–48 °C on the chlorophyll a fluorescence transients was the appearance of a very clear K step at 200–300 μs for both peel types. The K step was slightly more pronounced in the sun-exposed peel than in the shaded peel, suggesting that the resistance of oxygen-evolving complex to heat stress is slightly lower in the sun-exposed peel than in the shaded peel. Minimal fluorescence (F_O), relative to the value at 25 °C, increased to a greater extent in the shaded peel than in the sun-exposed peel after exposure to 46–48 °C, but the temperature dependencies of F_O changes were similar for both peel types. Maximum quantum yield of PSII (F_V/F_M) decreased to a similar extent in the sun-exposed peel and the shaded peel as temperature rose from 25 to 44 °C, but the sun-exposed peel reached slightly lower values at 46–48 °C. Correspondingly, gross O₂ evolution rate, relative to that at 25 °C, was also slightly lower in the sun-exposed peel than in the shaded peel at 46–48 °C. In response to heat stress, the ratio of Q_A-reducing reaction centers (RCs) to total RCs and the ratio of Q_B-reducing RCs to Q_A-reducing RCs decreased, but both of them decreased to lower values in the sun-exposed peel than in the shaded peel at 46–48 °C, indicating that the capacity of electron transfer between P₆₈₀⁺ and Q_B via Q_A was damaged to a greater extent in the sun-exposed peel than in the shaded peel. At each given temperature, dark respiration was similar between the two peel types. Overall, it appears that the exposure to higher surface temperature under high light does not make the sun-exposed peel more tolerant of heat stress than the shaded peel of apple fruit.     

Characterization of recombinant antibodies for cancer therapy by infrared spectroscopy

Fourier transform infrared (FTIR) spectroscopy was used to study the structure of the recombinant antibodies 1E10, anti-CD20 and hR3, which are used as anti-cancer therapeutic drugs. We tested their sensitivity against different conditions and treatments such as pH, temperature, freeze-thaw cycles and drying, which are relevant for the practical usefulness of the drugs. All antibodies were stable against moderate temperature increases (up to 50 °C) and pH changes (range 5–9). 1E10 was sensitive to extreme pH values (pH 3 and 12), whereas hR3 was most sensitive to temperature (at and above 60 °C). We did not observe any significant changes upon freeze-thaw and drying treatments. The secondary structure content of all three antibodies was estimated to be similar to that of IgG with ～64% β-sheet, 0% α-helix and ～36% other structure.     

Effects of heat stress on some reproductive parameters of male cavie (Cavia porcellus) and mitigation strategies using guava (Psidium guajava) leaves essential oil

Climate changes, particularly the increase of temperature are among the main causes behind the decline of fertility in humans as well as animals. In this study, the effects of heat stress on some reproductive parameters of male cavies and mitigation strategies using guava leaves essential oil (GLEO) were studied. For this purpose, 40 male cavies aged 2.5–3 months and weighing between 348 and 446 g were divided into 4 groups of 10 animals each and subjected to the following temperatures: Ambient temperature (20–25 °C) for the control group, 35 °C for group 1, 45 °C for group 2 and 45 °C+100 µl GLEO/kg body weight, administered by gavage to animals for group 3. Exposure time of heat was 7 h per day for 60 days. Results reveal that the relative weights of testes, epididymis, vas deferens and seminal vesicles were hardly affected by the temperature levels considered (P>0.05). The mass and individual sperm motility was significantly lower (P<0.05) in cavies exposed to the temperature of 35 and 45 °C as compared with those which received GLEO and controls. The percentages of abnormal sperm and altered sperm DNA were higher in animals exposed to temperature of 35 and 45 °C as compared with the controls. The activity of superoxide dismutase significantly increased (P<0.05) in animals exposed to temperature of 45 °C and in those of 45 °C and orally treated with GLEO, compared with cavies exposed to temperature of 45 °C without receiving GLEO. The level of malondialdehyde was significantly increased (P<0.05) in animals exposed to temperature of 35 and 45 °C, whereas the level of nitric oxide was significantly lower (P<0.05) in exposed animals as compared with controls. It was concluded that the exposure of male cavies at 35 and 45 °C for 60 days induce heat stress that causes deterioration of sperm characteristics. These effects that can be mitigated by the administration of guava leaves essential oil.     

Ultrasound-assisted extraction of some branded tea: Optimization based on polyphenol content,antioxidant potential and thermodynamic study

Tea is one of the top beverages used around the world every day, which contains a high amount of polyphenols and antioxidants. The main aim of this research is to quantify some marketed black tea (Rabea, Lipton, Alkbous, Green gold and Haritham) for phenolic contents and antioxidant potential evaluation by ultrasound solvent extraction and was compared with conventional extraction. Ultrasonic extraction was optimized by considering frequencies (26 kHz, 40 kHz), temperature (30, 40 and 50 °C), and power (30, 40 and 50%) at a fixed time of 30 min. In both the ultrasonic frequencies, 40 °C temperature and 40% power combination exhibited highest cumulative yield (mg/100 g DW), total phenolic content (mg gallic acid/g DW), flavonoids (mg/g DW) and DPPH radical scavenging activity (%) in all branded tea. Within each brand of tea, at any temperature-power combination at particular frequency results were not significantly different. But, at a similar condition of temperature power results were found significantly different between two frequencies. Furthermore, ultrasonic extraction process was analyzed thermodynamically by selecting some basic parameters. Thermodynamics results showed the extraction process was feasible, spontaneous and irreversible. Also, 26 kHz ultrasonic probe is more appropriate for the extraction purpose and thermodynamically more acceptable as compared to 40 kHz ultrasonic bath. Moreover, Haritham was selected as the best tea brand due to its high polyphenol contents and antioxidant potential.     

Variation in polyphenolic profile and in vitro antioxidant activity of eastern teaberry (Gaultheria procumbens L.) leaves following foliar development

Seasonal dynamics in the polyphenolic composition, antioxidant activity, and their relationships during plant development were evaluated for eastern teaberry (Gaultheria procumbens L.) leaves, a traditional herbal medicine of North American natives. With the complementary UHPLC-PDA-ESI-MS³, HPLC-PDA-fingerprint, Folin-Ciocalteau, and n-butanol/HCl assays of methanol-water (75:25, v/v) extracts, the dried leaf samples harvested monthly across the growing season under Polish climate conditions were found rich in structurally diverse polyphenols (149.2–210.7 mg/g DW) including the dominating salicylates (64.6–107.5 mg/g DW), proanthocyanidins (53.0–66.8 mg/g DW), and flavonoids (17.3–25.3 mg/g DW), and the accompanying chlorogenic acid isomers (2.4–4.4 mg/g DW) and simple phenolic acids (0.9–1.1 mg/g DW). Among 28 detected analytes, gaultherin (64.6–107.5 mg/g DW), miquelianin (14.6–21.1 mg/g DW), procyanidin A-type trimer (5.5–9.5 mg/g DW), and (–)-epicatechin (5.8–7.8 mg/g DW) were the most abundant. The phenolic levels and antioxidant activity parameters in the DPPH (EC₅₀, 15.0–18.2 μg DW/mL; 0.95–1.16 mmol Trolox equivalents/g DW) and FRAP (2.3–3.4 mmol Fe ²⁺/g DW; 0.86–1.26 mmol Trolox equivalents/g DW) assays showed parallel seasonal trends with maxima in September and October. As the subsequent correlation studies confirmed the determinative impact of polyphenols on the leaf antioxidant activity and its seasonal fluctuations, the Fall season could be recommended as optimal for harvesting the plant material for medicinal purposes and cost-effective production of natural health products.