Antiradical activity of different parts of Walnut (Juglans regia L.) fruit as a function of genotype

The objective of this work was to analyse phenolic compounds and antiradical capacity of different parts of walnut fruit among six genotypes of Juglans regia L. Therefore, total phenolic and flavonoid content were determined and methanolic extracts of walnut genotypes were considered by the reducing power, DPPH (2,2-diphenyl-1-picrylhydrazyl), superoxide anion and nitric oxide radical scavenging. Significant differences were found in phenolic content and radical scavenging capacity of different parts of fruits and among various genotypes. High correlation coefficient (R² = 0.81) was observed between phenol content and radical scavenging activity, but this was not always true (R² = 0.01). These results demonstrated that walnut genotypes have different phenolic compounds and phenolic compounds have different radical scavenging power. The differences of phenolic compounds were confirmed by using high performance liquid chromatography (HPLC).     

Evaluation of antioxidant ability of ethanolic extract from dill (Anethum graveolens L.) flower

Antioxidant activities of ethanolic extract from dill flower and its various fractions were evaluated with 2,2-diphenyl-1-picrylhydrazyl radical scavenging, Trolox equivalent antioxidant capacity, reducing power, chelating power, and β-carotene bleaching assays. The flower extract was successively separated into n-hexane, ethyl acetate and ethanol soluble fractions by liquid–liquid partition. Dill leaf and seed extracts were used for comparison. In all assays, the flower extract showed higher antioxidant activity than the leaf and seed extracts. With regard to various fractions of the flower extract, the sequence for antioxidant activity was ethyl acetate fraction > ethanol fraction > original flower extract > n-hexane fraction. Phenols including flavonoids and proanthocyanidins should be responsible for antioxidant abilities of the flower extract. Chlorogenic acid, myricetin, and 3,3’,4′,5,7-pentahydoxyflavan (4 → 8)-3,3′,4′,5,7-pentahydoxyflavan were the major phenolic acid, flavonoid, and proanthocyanidin, respectively, in the dill flower extract.     

In Vitro Antioxidant and Antiglycation Properties of Methanol Extract and Its Different Solvent Fractions of Musa paradisiaca L. (Cv. Nendran) Inflorescence

Methanolic extract of Musa paradisiaca inflorescence and its different solvent fractions, petroleum ether, ethyl acetate, butanol, and water were studied for their total phenolic and flavonoid content, free radical scavenging, and antiglycation activities, and these properties were compared with standard antioxidant compounds. Ethyl acetate fraction exhibited higher antioxidant and antiglycation properties than other fractions. IC₅₀ values of ethyl acetate fraction for 1,1-diphenyl-2-picrylhydrazyl method, 2,2-azinobis-3 ethylbenzothiazoline-6-sulphonic acid method, superoxide radical scavenging, hydroxyl radical scavenging, nitric oxide radical scavenging, and antiglycation activities were 9.80, 13.50, 26.40, 19.71, 25.73, and 31.00 μ g/ml, respectively. Total phenolic content of ethyl acetate (21.52 mg GAE/g) was significantly higher than other fractions. There was positive linear correlation between antioxidant activity and total phenolic content.      

Chemical composition and antioxidative activity of Moldavian balm (Dracocephalum moldavica L.) extracts

Moldavian balm (Dracocephalum moldavica L., Lamiaceae) is a perennial herb native to central Asia and naturalized in eastern and central Europe. It is commonly consumed as a food-related product and as a herbal preparation because of its reputed medicinal properties. Despite its importance, few reports exist in the literature regarding the chemistry or antioxidant activity of this species. In this study, the aerial material of Moldavian balm collected from Iran was extracted by Soxhlet using seven solvents of different polarity, viz., petroleum ether, dichloromethane, acetonitrile, ethyl acetate, methanol, n-butanol and water. The qualitative-quantitative chemical composition of each extract was determined using high-performance liquid chromatography coupled to photodiode array detection. For each extract, the total phenolic content was estimated as was the in vitro antioxidant activity using the iron(III) reduction assay, the β-carotene-linoleic acid bleaching assay and the 1,1-diphenyl-2-picrylhydrazyl and 2,2′-azinobis(3-ethylbenzothiazoline-6-sulphonate) free radical scavenging assays. Hydroxylated cinnamic acids, their derivatives and flavonoids were identified and quantified within the extracts, with rosmarinic acid being the most abundant component identified. The extracts demonstrated different degrees of potency within each assay, however, the observed pattern was not necessarily replicated between assays indicating the importance of the use of more than one screening technique to estimate the antioxidant activity of plant extracts.     

Antioxidant activities and related polyphenolic constituents of the methanol extract fractions from Broussonetia papyrifera stem bark and wood

Broussonetia papyrifera radix, fruits, leaves, and stems exhibit antioxidant, antinociceptive, antityrosinase, anti-inflammatory, and antiplatelet activities. However, study of the antioxidant activity of stem bark has been limited. In this work the antioxidant activity of stem bark and wood extracts were compared using different methods, and their phenolic constituents were analyzed. The bark ethyl acetate fraction exhibited the highest 2,2-diphenyl-2-picrylhydrazyl hydrate (DPPH) radical, hydroxyl radical scavenging activity, and the highest total phenolic and flavonoid contents. Both bark and wood n-butanol fractions showed stronger superoxide anion radical scavenging activity and reducing potential than other fractions. The bark and wood hexane fraction provided the highest level of ion chelating power. The high performance liquid chromatography (HPLC) analysis reveals there are more phenolic compounds in bark than in wood. These findings suggest that bark exhibits a higher antioxidant capacity than wood and that ethyl acetate and n-butanol fractions of B. papyrifera bark are potential natural resources for pharmacology of functional foods.     

Free radical scavenging activities and bioactive substances of Jerusalem artichoke (Helianthus tuberosus L.) leaves

The total phenolic content and radical scavenging activities of Jerusalem artichoke (Helianthus tuberosus L.) leaves were investigated. Results indicated that the ethyl acetate fraction contained the highest total phenolic content (266.69 ± 2.51 mg GAE/g dry extract) accompanied with strongest free radical scavenging abilities. Following an in vitro radical scavenging activity-guide fractionation procedure, six phenolic compounds which strongly quenched free radicals were separated from ethyl acetate fraction. Among them, 3-O-caffeoylquinic acid and 1,5-dicaffeoylquinic acid played a dominant role due to their strong free radical scavenging abilities and their high contents. The content of 3-O-caffeoylquinic acid in n-butanol fraction was 74.58 ± 1.05 mg/g, while 1,5-dicaffeoylquinic acid in ethyl acetate fraction was 104.51 ± 2.86 mg/g. The results imply that the leaves of Jerusalem artichoke might be a potential source of natural antioxidants.     

Survey of 55 Turkish Salvia taxa for their acetylcholinesterase inhibitory and antioxidant activities

In the current work, our target was to screen inhibitory potentials of 55 Turkish Salvia taxa, 28 of which are endemic, against acetylcholinesterase (AChE), which is a chief enzyme in pathogenesis of Alzheimer’s disease (AD). The dichloromethane, ethyl acetate, and methanol extracts prepared from 55 Salvia taxa were tested for their AChE inhibitory activity at 25, 50, and 100 μg/ml using an ELISA microplate reader. The extracts were also screened for their scavenging effect against DPPH radical and iron-chelating capacity. Total phenol and total flavonoid contents of Salvia fruticosa were determined. Among the 165 Salvia extracts screened, only the dichloromethane extract of S. fruticosa showed inhibition towards AChE at 100 μg/ml having 51.07% of inhibition, while only the dichloromethane and ethyl acetate extracts of Salvia cilicica had a notable iron-chelating capacity at 100 μg/ml having 54.71% of chelating capacity. Most of the extracts showed remarkable scavenging effect against DPPH radical.     

Evaluation of antioxidant activities of various solvent extracts of Carissa opaca fruits

The chloroform and aqueous fractions of Carissa opaca fruit, a traditional medicinal fruit in Pakistan possessed a high amount of total phenolic and flavonoid contents as compare to other solvent fractions with potent antioxidant activities in scavenging DPPH, superoxides, hydroxyl, hydrogen peroxide, ABTS radicals, and had strong iron chelating activity. On the other hand, the ethyl acetate fraction showed the highest inhibition of β-carotene linoleic acid peroxidation and phosphomolybdate assay. A high correlation coefficient existed between EC₅₀ values of DPPH, superoxides, hydroxyl, hydrogen peroxide, ABTS radicals, total phenolics and flavonoids, but a non significant correlation was found in the case of iron chelaters, β-carotene and phosphomolybdate assay. This study verified that the chloroform and aqueous fractions have strong antioxidant activities which were correlated with its high level of phenolics and flavonoids. These fractions can be used as a source of potential antioxidant or functional food material.     

Assessment of antiradical potential of Calluna vulgaris (L.) Hull and its major flavonoid

BACKGROUND: Antioxidant capacity of the chloroform, ethyl acetate, n‐butanol and water fractions of the aerial parts of Calluna vulgaris (L.) Hull (Ericaceae) has been assessed in this study. Antioxidant capacity of the plant was screened by assays of 2,2‐diphenyl‐β‐picrylhydrazyl, superoxide anion and hydrogen peroxide scavenging, metal‐chelating activity and reducing power. Butylated hydroxyanisole was used as reference in all assays; ethylene diamine tetraacetic acid was also used as reference in the assay of metal‐chelating activity. Total phenolic contents of the fractions were determined by the Folin–Ciocalteu method. RESULTS: Liquid chromatography/diode array detection/mass spectrometry was used for phytochemical identification of the fractions. Kaempferol‐3‐O‐β‐D ‐galactoside was found to be the major constituent in the ethyl acetate fraction (37.1 ± 0.9%), followed by the n‐butanol fraction (4.6 ± 0.1%). High occurrence of antioxidant capacity, with the exception of metal‐chelating activity, was observed in the ethyl acetate and chloroform fractions as well as in kaempferol‐3‐O‐β‐D ‐galactoside. CONCLUSION: Calluna vulgaris and its major flavonoid, kaempferol‐3‐O‐β‐D ‐galactoside, show high antioxidant capacity in various assays. As far as is known, this is the first report on antioxidant capacity of C. vulgaris and its major flavonoid. Copyright © 2009 Society of Chemical Industry     

Anticholinesterase and antioxidant effects of the ethanol extract, ethanol fractions and isolated flavonoids from Cistus laurifolius L. leaves

In the current study, the n-hexane, chloroform, ethyl acetate, water, and n-butanol fractions obtained from the main ethanol extract of Cistus laurifolius L. were evaluated for their cholinesterase inhibitory effects against acetyl- (AChE) and butyrylcholinesterase (BChE), at 50, 100, and 200 μg/ml, using an ELISA microplate reader. The antioxidative effect of the extract and fractions was also determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, ferrous ion-chelation capacity, and ferric-reducing antioxidant power (FRAP) test systems. Total phenol and flavonoid contents of the extract and fractions were calculated using Folin-Ciocalteau and aluminium chloride reagents. Three flavonoid derivatives; 3-O-methylquercetin (1), 3,7-O-dimethylquercetin (2), and 3,7-O-dimethylkaempferol (3) isolated from the CHCl₃ fraction were also tested in the same manner. Our experimental findings indicated that the ethanol extract exerted the highest AChE inhibition (80.07 ± 1.06% at 200 μg/ml). The ethyl acetate and n-butanol fractions displayed the best activity against DPPH and FRAP assays.     

Antioxidant capacity of pummelo and navel oranges: Extraction efficiency of solvents in sequence

Extraction efficiency of five different solvents on the antioxidant capacities of pummelo and navel oranges was measured by five widely used antioxidant methods. Freeze-dried edible parts of pummelo and navel oranges were extracted with five different solvents such as hexane, ethyl acetate, acetone, methanol and methanol:water (8:2). The total phenolic content of the extracts was determined by Folin-Ciocalteu method and expressed as catechin equivalents. The highest phenolic content was obtained from ethyl acetate and the minimum phenolic content was found in methanol extract. The dried extracts were screened for their radical scavenging activity using three different methods such as α,α-diphenyl-β-picrylhydrazyl (DPPH), 2,2′-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical assay and ORAC methods. Ethyl acetate extract from navel orange and pummelo was found to be most active radical scavenging activity, whereas hexane extract from pummelo and methanol extract from navel orange was found to be lowest activity. Moreover, all the extracts from pummelo and navel orange were studied for their antioxidant capacity by the formation of phosphomolybdenum complex and reducing power by potassium ferricyanide reduction method at various concentrations. The order of antioxidant capacity of pummelo and navel orange was found to be ethyl acetate>acetone>MeOH:water>methanol> hexane and ethyl acetate>MeOH:water>acetone>methanol>hexane, respectively. Acetone and methanol extracts from pummelo and navel oranges showed highest reducing power than other extracts at 1000 μg/ml. Significant differences in antioxidant capacity were found between the values obtained by the same method in different solvents and as well as each extract antioxidant capacity obtained by the different method. Results of the present study may be due to the extent of antioxidant capacity of each extract is in accordance with the amount of carotenoids, phenolics and vitamin C present in the extracts.     

Free radical scavenging activity and phenolic content in achenes and thalamus from Fragaria chiloensis ssp. chiloensis, F. vesca and F. x ananassa cv. Chandler

The total phenolic, flavonoid and anthocyanin content of achenes (true fruit) and thalamus (receptacle) from the native South American Fragaria chiloensis ssp. chiloensis (f. patagonica and f. chiloensis), Fragaria vesca and Fragaria x ananassa cv. Chandler was determined by spectrophotometric means. Highest phenolic content was found in F. vesca while lowest content was measured for white strawberry (F. chiloensis ssp. chiloensis, f. chiloensis). The total anthocyanin and total flavonoid contents in the samples investigated was lower for the white strawberry and higher in F. x ananassa cv. Chandler. Total flavonoid content showed a better correlation than total anthocyanins with the free radical scavenging effect of the extracts measured by means of the DPPH discoloration assay. In the superoxide anion assay all the acetone extracts of strawberries showed similar activity. The data presented in this study demonstrate that the amount of phenolic compounds differ significantly between species and subspecies and determine the free radical scavenging activity of fruits. On a w/w basis, higher total phenolics including flavonoids was found in achenes. The highest total anthocyanin content was found in the achenes of F. chiloensis and F. vesca, while F. ananassa presented higher antocyanin content in thalamus. The main anthocyanin in thalamus of F. ananassa (95%) were pelargonidin derivatives which were also present in F. chiloensis ssp. chiloensis f. patagonica (62.6%) but were not detected in F. vesca and F. chiloensis ssp. chiloensis f. chiloensis. In F. vesca and F. chiloensis ssp. chiloensis f. chiloensis only cyanidin derivatives were found both in thalamus and achenes.     

Studies on antioxidant potential of methanol extract/fractions of Acacia auriculiformis A. Cunn

Antioxidant activities of the methanol extract/fractions of Acacia auriculiformis A. Cunn were evaluated by three in vitro experiments, namely, DPPH, relative reducing power and hydroxyl radical (site specific and non site specific) assays. The differential activities of methanol extract/fractions could be correlated with their respective total phenolic contents and compared with standards (BHT and l-ascorbic acid.). The bark powder of the plant was extracted with different solvents of increasing and decreasing polarity by a maceration extraction method and then the methanol extract was further partitioned with ethyl acetate and water. The scavenging activity of extract was found to be enhanced on fractionating the extract. Moreover, among the two fractions (ethyl acetate and water fraction) and the crude extract, the water fraction exhibited good scavenging responses of 72.0%, (57.2%), 1.76 (1.52), 88.0% (82.6%) and 93.6% (83.47) in DPPH, reducing power, site specific and non-site specific hydroxyl radical scavenging assay in increasing and (decreasing) order of solvent polarity at maximum concentration, respectively. Studies are in progress to evaluate the effect of extracts/fractions in other antioxidant assays and to identify the factors responsible for the activity.     

In vitro scavenging capacity of annatto seed extracts against reactive oxygen and nitrogen species

Bixa orellana L. (annatto), from Bixaceae family, is a native plant of tropical America, which accumulates several carotenoids (including bixin and norbixin), terpenoids, tocotrienols and flavonoids with potential antioxidant activity. In the present study, the in vitro scavenging capacity of annatto seed extracts against reactive oxygen species (ROS) and reactive nitrogen species (RNS) was evaluated and compared to the bixin standard. Annatto extracts were obtained using solvents with different polarities and their phenolic compounds and bixin levels were determined by high performance liquid chromatography coupled to diode array detector. All annatto extracts were able to scavenge all the reactive species tested at the low μg/mL range, with the exception of superoxide radical. The ethanol:ethyl acetate and ethyl acetate extracts of annatto seeds, which presented the highest levels of hypolaetin and bixin, respectively, were the extracts with the highest antioxidant capacity, although bixin standard presented the lowest IC₅₀ values.     

Assessment of the antioxidant potential and fatty acid composition of four Centaurea L. taxa from Turkey

This paper focused on the assessment of antioxidant property and fatty acid composition of four Centaurea species. The antioxidant activity of its methanol extract was evaluated by several in vitro experiments including phosphomolybedum assay, DPPH assay, β-carotene/linoleic acid, ferric and cupric reducing power. Total phenolic and flavonoid contents were also evaluated. The methanol extract of Centaurea pulcherrima var. pulcherrima showed the superior free radical scavenging activity, linoleic acid inhibition capacity, reducing power and also had the highest total phenolic content. A significant relationship between antioxidant capacity and total phenolic components was found. The oils of Centaurea taxa were also analysed for fatty acid concentration by gas chromatography. The principal fatty acids in the species were palmitic acid (23.38–30.49%) and linoleic acid (20.19–29.93%). These findings suggest that the Centaurea species could be used as a potential source of new natural antioxidants and unsaturated fatty acids in food industry, cosmetics and pharmaceutical preparations.     

Biological activities of flavonoids from Nitraria retusa (Forssk.) Asch. and their acylated derivatives

The antioxidant activity and antiproliferative property towards Caco2 cells of water and methanol extracts/fractions of Nitraria retusa were investigated. The total phenolic and flavonoid contents of the extracts/fractions were determined, and the four major flavonoids were identified as isorhamnetin, isorhamnetin-3-O-glucoside, isorhamnetin-3-O-rutinoside and isorhamnetin-3-O-robinobioside. The results showed a relationship between the extracts/fractions activities and their flavonoid contents. Moreover, the chloroform extract which was enriched with the aglycone flavonoid isorhamnetin exhibited the highest activities. The activities of N. retusa flavonoids were compared to those of model flavonoids, quercetin, isoquercitrin and rutin. In all cases, the aglycone compounds were more active than their glycosylated derivatives. Isorhamnetin-based flavonoids presented higher antiproliferative activities than quercetin-based ones, while similar antioxidant properties were observed. The enzymatic acylation of isorhamnetin-3-O-glucoside with ethyl laurate and ethyl butyrate enhanced its capacity to inhibit xanthine oxidase and its antiproliferative activity but decreased its radical scavenging activity.     

Antioxidant phenolic compounds from Smilax sebeana Miq.

Methanol extract and sub-fractions of Smilax sebeana rhizomes and roots were analyzed to evaluate the compounds involved in antioxidant activity. In all separated fractions of the different polarity solvents, ethyl acetate fraction showed the highest antioxidant activity and total phenolic content. This fraction was subjected to the sephadex LH-20 column and preparative HPLC for purification. Six phenolic compounds, chlorogenic acid (1), 4-formylphenol (2), epicatechin (3), cinchonain IIa (4), Ia (5) and Ib (6) were isolated and identified by spectroscopic analyses and further evaluated their potential antioxidant activities by DPPH and superoxide radical scavenging assays. Compared with synthetic antioxidant Trolox, except 4-formylphenol, the other isolated five compounds exhibited excellent antioxidant activities. This is the first report on the chemical constituents of S. sebeana which potentially involved in antioxidant activity. The results suggest that the ethyl acetate extract of S. sebeana might be used as a readily accessible source of natural antioxidant.     

Cyphomandra betacea Sendt. phenolics protect LDL from oxidation and PC12 cells from oxidative stress

Cyphomandra betacea Sendt. (tamarillo) is a subtropical fruit containing rich contents of anthocyanins and carotenoids. The antioxidant activity was investigated using a crude ethanol extract of C. betacea fruit and its partitioned fractions, i.e. the ethyl acetate, butanol and water fractions. The ethyl acetate fraction exhibited the highest DPPH scavenging activity, Trolox equivalent antioxidant capacity (TEAC) and total phenol content. C. betacea phenolics in ethyl acetate fraction inhibited copper-induced LDL oxidation equally to or more effectively than dl-α-tocopherol, as measured by decreased formation of thiobarbituric acid-reactive substances (TBARS), conjugated diene and relative electrophoretic mobility (REM). Furthermore, 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium (MTT) reduction assay showed that C. betacea phenolics in ethyl acetate fraction prevent oxidative stress-induced cell death in neuronal PC12 cells in a dose-dependent manner via attenuation of ROS production. In conclusion, C. betacea phenolics are potent antioxidants which can inhibit LDL oxidation in vitro and ROS production in PC12 cells.