Comparative Study on the Antioxidant Potential of Milk Thistle Seeds and Silymarin Extract: Potential Implications in Ruminants

Achillea is a traditional medicinal herb which contains different phenol and flavonoid compounds that are responsible for Achillea pharmacological effects. We aimed to determine phenol and flavonoid contents, besides antioxidant activities of different extracts from Achillea eriophoraa (A. eriophora) DC. and Achillea biebersteinii (A. biebersteinii) Afan. (endemic species in Iran) and to investigate their effects on human cells. Achillea extracts, were prepared by maceration and shaking methods, from different parts (aerial parts, stem, leaves and inflorescence) of two species using methanol and ethanol as solvents. Total phenol and flavonoid contents were measured by spectrophotometry, and antioxidant activities of the extracts were determined by DPPH radical scavenging, BCB and TBARS assays. Cytotoxicity and antioxidant activities of the extracts were investigated in Human Foreskin Fibroblast (HFF3) cells using MTT, comet and H2O2 assays. Methanol extracts of A. biebersteinii prepared from leaves and inflorescence by maceration method exhibited maximum phenol (1657.58 ± 36.45 mg GAE/100 g DW) and flavonoid (264.00 ± 62.16 mg QUE/100 g DW) contents. Leaf methanol extract showed significantly higher antioxidant activity (0.0276 ± 0.003, 0.16 ± 0.016 and 13.96 ± 0.26 mg/ml for DPPH, BCB and TBARS IC50s, respectively) than those of the other extracts. Leaf extract of A. biebersteinii was not cytotoxic even at the highest examined dose (512 µg/ml) and inhibited cell toxicity induced by H2O2 (98% viability for the cells pretreated with plant extract in the presence of H2O2). Comet assay also confirmed high DNA protective activity of leaf extracts. Achillea extracts possess remarkable antioxidant activity, and could be good natural alternatives to synthetic antioxidants in pharmaceutical and food industries.

Black cumin (N. sativa; Ranunculaceae family) is well known for its numerous beneficial biological effects, while its seed extracts exhibit anti-diabetic, anti-cancer, immunomodulatory, anti-microbial, anti-inflammatory, anti-hypertensive and antioxidant activities [1,2]. This study aimed to evaluate the phenolic and mineral contents, chemical composition and antioxidant activity of methanolic extracts from dried black cumin seed powder. In addition, we evaluated, after gastrointestinal digestion, the effect of phenolic components and their antioxidant activity by utilising an in vitro gastrointestinal digestion process. Black cumin showed high amounts of total phenolic and flavonoid contents (such as dihydroxybenzoic acid and ferulic acid) via HPLC analysis. Six mineral elements (Ca, Cu, Fe, K, Se and Zn) were determined by using coupled plasma mass spectrometry. Twenty-five (25) fatty acids (13 saturated, 7 unsaturated and 5 unsaturated omega fatty acids) were identified by gas chromatography, with linoleic acid being the most abundant. In addition, black cumin methanolic extract presented higher antioxidant capacity measured by DPPH, FRAP and TEAC. Finally, dried black cumin powder was evaluated after gastrointestinal digestion, with results indicating that phenolics, flavonoids and antioxidant capacity increased in the gastric fraction (1.81-, 1.03- and 2.1-fold, respectively) compared with the undigested methanolic extract. Moreover, a higher amount of total phenolic and flavonoid content, as well as a higher total antioxidant capacity were found present in the gastric and elimination fraction than in the bio-accessible fraction (that represent the colon availability). Our results demonstrate a significant reduction in the quantity of phenolic (68%) and flavonoid (95.53%) compounds after gastrointestinal digestion in the bio-accessible fraction, together with a decrease in total antioxidant activity, suggesting that phenolic compounds are responsible for the observed antioxidant activity.

BackgroundThe aim of this study was to screen various solvent extracts of whole plant of Torilis leptophylla to display potent antioxidant activity in vitro and in vivo, total phenolic and flavonoid contents in order to find possible sources for future novel antioxidants in food and pharmaceutical formulations.Material and methodsA detailed study was performed on the antioxidant activity of the methanol extract of whole plant of Torilis leptophylla (TLM) and its derived fractions {n-hexane (TLH), chloroform (TLC) ethyl acetate (TLE) n-butanol (TLB) and residual aqueous fraction (TLA)} by in vitro chemical analyses and carbon tetrachloride (CCl4) induced hepatic injuries (lipid peroxidation and glutathione contents) in male Sprague-Dawley rat. The total yield, total phenolic (TPC) and total flavonoid contents (TFC) of all the fractions were also determined. TLM was also subjected to preliminary phytochemical screening test for various constituents.ResultsThe total phenolic contents (TPC) (121.9±3.1 mg GAE/g extract) of TLM while total flavonoid contents (TFC) of TLE (60.9 ±2.2 mg RTE/g extract) were found significantly higher as compared to other solvent fractions. Phytochemical screening of TLM revealed the presence of alkaloids, anthraquinones, cardiac glycosides, coumarins, flavonoids, saponins, phlobatannins, tannins and terpenoids. The EC50 values based on the DPPH (41.0±1 μg/ml), ABTS (10.0±0.9 μg/ml) and phosphomolybdate (10.7±2 μg/ml) for TLB, hydroxyl radicals (8.0±1 μg/ml) for TLC, superoxide radicals (57.0±0.3 μg/ml) for TLM and hydrogen peroxide radicals (68.0±2 μg/ml) for TLE were generally lower showing potential antioxidant properties. A significant but marginal positive correlation was found between TPC and EC50 values for DPPH, hydroxyl, phosphomolybdate and ABTS, whereas another weak and positive correlation was determined between TFC and EC50 values for superoxide anion and hydroxyl radicals. Results of in vivo experiment revealed that administration of CCl4 caused a significant increase in lipid peroxidation (TBARS) while decrease in GSH contents of liver. In contrast, TLM (200 mg/kg bw) and silymarin (50 mg/kg bw) co-treatment effectively prevented these alterations and maintained the antioxidant status.ConclusionData from present results revealed that Torilis leptophylla act as an antioxidant agent due to its free radical scavenging and cytoprotective activity.

The nutritional composition and antioxidant activity (in aqueose and solvent extracts) of two medicinal plants of Iranian origin Borage (Echium amoenum) and Valerian (Valerian officinalis) used as tea were determined. Samples were analyzed for antioxidant components viz. polyphenols, vitamin C, β carotene, flavonoids, anthocyanins and tannins. Antioxidant assays such as free radical scavenging activity, reducing power and total antioxidant activity were carried out for ethanol, methanol, acetone, 80% methanol and 80% ethanolic extracts. In borage highest and least activity was observed in water and acetone extract respectively in all assays. In Valerian, 80% methanolic extract showed highest activity in reducing power and free radical scavenging activity assay. Total polyphenols in borage and valerian were 1,220 and 500mg in ethanolic extracts and 25 and 130mg in acetonic extracts respectively. Total carotenoids and vitamin C contents were 31.6 and 133.69mg and 51.2 and 44.87mg for borage and valerian respectively. Highest amount of tannins were extracted in 80% methanolic extract. It can be concluded that borage and valerian exhibited antioxidant activity in all extracts. The antioxidant activity could be attributed to their polyphenol and tannin and flavonoids contents. In all assays borage showed higher activity than valerian.

Ficus sur is employed widely in folklore medicine for the mitigation of several disease conditions. The study aims to validate the phytochemical composition, antioxidant, and anti-inflammatory activities of hydroethanolic extract and fractions (dichloromethane, ethyl acetate, and n-butanol) of F. sur stem bark. Phytochemical profiling of the extract and fractions was carried out by established methods and gas chromatography-mass spectroscopy (GC-MS) technique, while their total phenolic (TPC) and flavonoid contents (TFC) were assessed by the Folin-Ciocalteu reagent and aluminium chloride complex formation methods, respectively. In vitro antioxidant activity was carried out by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging, ferric reducing antioxidant power (FRAP), and total antioxidant capacity (TAC) assays. The carrageenan-induced paw oedema protocol was employed in the evaluation of the anti-inflammatory activity of the extract and fractions. Phytochemicals such as terpenoids, phenolics, flavonoids, and glycosides were found to be present in the extract and fractions. However, GC-MS analysis of the extract revealed the presence of majorly oleanane-type and phytosterol triterpenoids, while polyphenolic compounds and fatty acids existed in small abundances. The extract and fractions showed considerably high TAC and TFC values, and substantial antioxidant activities in the DPPH, FRAP, and TAC antioxidant assays. The groups treated with n-butanol fraction and the standard drug (diclofenac) exhibited the highest anti-inflammatory effects with maximum percentage inhibitions of 62.05% and 75.63% of paw oedema in the rats, respectively. The study revealed that F. sur stem bark possessed rich content of triterpenoids and polyphenols, and demonstrated considerable antioxidant and anti-inflammatory properties as suggested by ethnomedicine.

This study aims to evaluate the phenolic contents, chemical composition, mineral amount as well as antioxidant activity of methanolic extracts from dried black cumin (Nigella sativa L; family: Ranunculaceae) seed powder. We also performed an in vitro gastrointestinal digestion process to understand the effect on phenolic components and their antioxidant activity after gastrointestinal digestion. Black cumin showed high amounts of total phenolic and flavonoid contents, such as dihydroxybenzoic acid and ferulic acid, through high‐performance liquid chromatography analysis. Six mineral elements (Ca, Cu, Fe, K, Se, and Zn) were determined by using coupled plasma mass spectrometry, while 25 fatty acids (13 saturated, 7 unsaturated, and 5 unsaturated omega fatty acids) were also identified by gas chromatography, with linoleic acid the most present. In addition, black cumin extract presented high antioxidant capacity measured by DPPH, FRAP, and TEAC. Additionally, dried black cumin powder was evaluated after gastrointestinal digestion: phenolic compounds (1.81‐fold), flavonoids (1.03‐fold), and antioxidant capacity (up to 2.1‐fold) increased in gastric fraction compared to the undigested methanolic extract. Moreover, a higher amount (p < 0.05) of total phenolic content and flavonoids, as well as higher total antioxidant capacity, were found in the gastric and elimination fraction than in the bioaccessible fraction. Our results demonstrated that there were a significant decrease in the quantity of phenolic compounds (68%) and flavonoids (95.53%) after gastrointestinal digestion in the bioaccessible fraction as well as a reduced the antioxidant activity (10.79%–94.84%), suggesting that phenolic compounds are responsible for antioxidant activity.

: Total phenolic and flavonoid contents in different extracts of Nepeta binaludensis Jamzad aerial parts were evaluated by colorimetric methods and the best results were found in 50% hydro-methanol extract with total phenolic (4.42 ± 0.0014 mg of gallic acid/g of dry mass of plant) and flavonoid (0.8 ± 0.0009 mg of quercetin/g of dry mass of plant) contents. Antioxidant activity of the extracts were also evaluated using 1,1-diphenyl-2-picrylhedrazyl (DPPH) assay. The best result was found in 50% hydro-methanol extract (the half-maximal inhibitory concentration IC50 = 3.0 mg/mL). A good correlation between the radical scavenging activity and total phenolic and flavonoid contents of the extracts was observed. Antimicrobial activity of 50% hydro-methanol and ethyl acetate extracts of the aerial parts of N. binaludensis were also evaluated on four different bacteria and a fungus in the current study. Considerable antibacterial activity against Staphylococcus aureus (4.0 mg/mL) and Streptococcus pyogenes (4.0 mg/mL) were observed for ethyl acetate extract in comparison with the other samples.

In vitro antioxidant studies of Begonia malabarica Lam. and Begonia floccifera Bedd

In traditional medicine, numerous diseases are treated using the leaves of Anthocleista vogelii. The total phenolic content and flavonoid content (antioxidants); and the effect of leaves extracts (ethanolic extract and fractions [n-hexane, dichloromethane and ethyl acetate]) of Anthocleista vogelii were determined in this study. The free radical scavenging property of the extracts were investigated using nitric oxide, hydrogen peroxide and 1, 1-Diphenyl-2-picrylhydrazyl assays. Spectrophotometric method was used for evaluating the antioxidant property of Anthocleista vogelii leaves. In this study, the results showed that Anthocleista vogelii leaves extracts exerted antioxidant and free radical scavenging activities. Total Phenol and flavonoid were present in the leaves extract. Ethyl acetate fraction exerted a more free radical scavenging effect more than other extracts of Anthocleista vogelii leaves. Ehtyl acetate fraction scavenged nitric oxide (68.19%), hydrogen peroxide (80.0%) and 1, 1-Diphenyl-2-picrylhydrazyl (92.02%). Ethyl acetate fraction also had the highest total antioxidant capacity (70.69 mg AAE/g), phenol (55.17 mg GAE/g) and flavonoid (60.23 mg QE/g) content than other extracts of Anthocleista vogelii leaves. The results from this study conclude that, Anthocleista vogelii leaves extracts (ethanolic extract and fractions [n-hexane, dichloromethane and ethyl acetate]) have antioxidant activities and ethyl acetate fraction exerted more antioxidant activity.

Summary Introductıon: Instead of synthetic antioxidants, using of natural products with antioxidant activity is demanded. Objectıve: The aim of this study is to evaluate the antioxidant activity and total phenolic and flavonoid contents of the Trachystemon orientalis (borage) obtained from Ordu. Methods: The water and ethanol extracts of the borage plant were investigated by using several in vitro antioxidant activity tests such as total antioxidant capacity, O2 •−, •OH, DPPH and ABTS radicals scavenging activities, hydrogen peroxide scavenging activity, FRAP, Fe2+ chelating and reducing power assays in order to make effective comparisons. Results: Water extract of the T. orientalis had the more phenolic (90 mg GA/g extract) and flavonoid (56.88 mg CT/g extract) contents than ethanol extract. Furthermore, both of the extracts showed strong antioxidant and radical scavenging activities determined by different in vitro models. The highest total antioxidant capacity as ascorbic acid equivalents of 244.50 mg/g extract was also calculated for water extract. Generally, a relationship between total phenolic content and antioxidant activity was established. It is also an important finding for an edible food source that the phenolic and flavonoid content and antioxidative activities for water extract was higher than the ethanol extract. Conclusıon: T. orientalis could be used as a food ingredient instead of synthetic antioxidants and all results will contribute to the recent increase in investigations on using natural products in many areas such as food, pharmacy, alternative medicine and natural therapy.

Capsicum annuum, a type of chilli, is a species that is extensively grown, a good source of bioactive chemicals, and an important component of diets that promotes health. Waste seeds from chilli processing have the potential to be used as an alternative source for antioxidants due to their chemical compositions, however, C. annuum seeds are still poorly studied. Thus, this study was carried out to evaluate the differences between C. annuum pericarp and seed extracts in terms of total phenolic and flavonoid contents and antioxidant activity. Antioxidant activity was measured using DPPH free radical scavenging assay. The pericarp extract exhibited a higher level of total phenolics (31.26 ± 6.71 μg GAE/mg dry weight) compared to the seeds (26.76 ± 2.08 μg GAE/mg dry weight) albeit not statistically significant. Similarly, a higher total flavonoid content was observed in the pericarp compared with the seed extract (21.65 ± 1.13 vs. 16.43 ± 0.37 μg QE/mg dry weight, p < 0.05). As a result, in comparison to the seed extract, the pericarp extract likewise demonstrated a greater DPPH radical scavenging activity. Despite having lower levels of total phenolics, total flavonoids, and DPPH scavenging activity than the pericarp, the seed extract is nevertheless significant and may be used as a substitute source of phenolics or other compounds rather than being thrown away as waste. This study promotes sustainability and the conversion of waste product into valuable resources of that may be used in the nutraceutical, pharmaceutical and food industry.

Investigation into the antioxidant activity and chemical composition of alcoholic extracts from defatted marigold (Tagetes erecta L.) residue

Many plants contain bioactive compounds such as polyphenols, flavonoids and tannins, which have antioxidant properties. Antioxidants help protect cells from damage caused by free radicals. The objective of this study was to investigate the antioxidant activity of plant extracts from Malcolmia aegyptiaca and Matthiola livida species. Aerial parts of the plants were collected and dried, and methanolic extracts were prepared from the dried plants. The total phenolic, flavonoid and tannin content of the extracts was determined and the antioxidant activity of the extracts was evaluated using various in vitro and in vivo assays. M. livida had a higher total phenolic and flavonoid content than M. aegyptiaca. Both extracts showed antioxidant activity in the assays, with M. livida generally exhibiting higher antioxidant activity than M. aegyptiaca. M. livida and M. aegyptiaca are both good sources of bioactive compounds with antioxidant activity. Therefore, these plants have potential for use in the development of natural antioxidants for the prevention and treatment of oxidative stress-associated diseases.

Microscopic fungi synthesize a wide range of phenolic compounds that play an important role in cell wall formation, as well as perform signaling and protective functions. One of the important functions of phenolic compounds, primarily polyphenols, is antioxidant activity. The most active polyphenols are flavonoids, their synthesis in microscopic fungi has not been studied enough. The aim of the study was to determine the total phenols and flavonoids content and antioxidant activity of Talaromyces funiculosus strains isolated from different econiches. Methods. 20 strains of the microscopic fungus T. funiculosus – 10 endophytes and 10 saprotrophs were used in the experiments. Cultivation of filamentous fungi was performed under submerged conditions with shaking at 210–230 rpm in Erlenmeyer flasks containing 200 ml of Czapek liquid medium at a temperature 26±2°C for 6 days. The biomass of the strains was filtered, dried at 70°C and extracted with 95% methanol (1:50 w/v). The total phenols content was determined using Folin-Ciocalteu reagent by the method of Arora, Chandra (2010) with some modifications and expressed as mg of gallic acid equivalents (GAE) per g of dry weight mycelium. Flavonoids in each extract were determined spectrophotometrically with AlCl3 and expressed as mg of quercetin equivalents (QE) per g of dry weight mycelium. Antioxidant activity of methanolic extracts was measured as radical scavenging activity by the percentage inhibition of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. Results. The total phenolic content of methanolic extracts of T. funiculosus strains ranged from 0.12±0.007 to 1.97±0.150 mg GAE/g. The highest level of phenolic compounds was found for saprotroph UCM F-16790 (1.97±0.150) and endophyte UCM F-16795 (1.71±0.137 mg GAE/g). The flavonoid content in methanolic extracts of the studied strains varied in the range of 0.02±0.001–0.39±0.041 mg QE/g. All studied strains of T. funiculosus showed antioxidant activity against DPPH radical, which was 12.19±2.34–60.37±7.24% inhibition. Statistical analysis revealed a strong positive correlation between the total phenols and flavonoids content (Pearson’s correlation coefficient r=0.92); between antioxidant activity, phenols (r=0.82) and flavonoids (r=0.81) content in methanol extracts of T. funiculosus strains. The total content of phenols and flavonoids and antioxidant activity did not depend on the trophic group of the studied strains. The moderate correlation between flavonoids content and antioxidant activity of methanolic extracts of endophytic T. funiculosus strains (r=0.46) may be related to their ability to synthesize a wider range of polyphenolic compounds with antioxidant activity. Conclusions. The studied T. funiculosus strains have the ability to synthesize phenolic compounds and flavonoids in different concentrations. Correlation analysis revealed strong correlations between total phenolic and flavonoid contents and antioxidant activity. The reason for phenomenon of moderate correlation (r=0.46) between flavonoid content and antioxidant activity in endophytic strains may be the synthesis of other compounds by individual strains. The determination of individual components synthesized by the most active strains is of scientific interest for further study. T. funiculosus UСM F-16795 and UСM F-16790 strains had a quite high level of phenols and flavonoids. Obtained data may be promising for the search for phenolic compounds with antioxidant properties to increase plant resistance to negative impacts of environmental stress factors.

The total phenolics and antioxidant activities of fenugreek, green tea, black tea, grape seed, ginger, rosemary, gotu kola, and ginkgo extracts, vitamin E, and tert-butylhydroquinone, were determined. Grape seed and green tea were analyzed for their phenolic constituents using high-performance liquid chromatography. The total phenolics of the plant extracts, determined by the Folin-Ciocalteu method, ranged from 24.8 to 92.5 mg of chlorogenic acid equivalent/g dry material. The antioxidant activities of methanolic extracts determined by conjugated diene measurement of methyl linoleate were 3.4-86.3%. The antioxidant activity of the extracts using chicken fat by an oxidative stability instrument (4.6-10.2 h of induction time) followed a similar trend in antioxidant activity as determined by the Folin-Ciocalteu method. Seven phenolics in grape seed and green tea extracts were identified that ranged from 15.38 to 1158.49 and 18.3 to 1087.02 mg/100 g of extract, respectively. Plant extracts such as green tea and grape seed extracts can be used to retard lipid oxidation in a variety of food products.