Biological activity of peanut (Arachis hypogaea) phytoalexins and selected natural and synthetic Stilbenoids

The peanut plant (Arachis hypogaea L.), when infected by a microbial pathogen, is capable of producing stilbene-derived compounds that are considered antifungal phytoalexins. In addition, the potential health benefits of other stilbenoids from peanuts, including resveratrol and pterostilbene, have been acknowledged by several investigators. Despite considerable progress in peanut research, relatively little is known about the biological activity of the stilbenoid phytoalexins. This study investigated the activities of some of these compounds in a broad spectrum of biological assays. Since peanut stilbenoids appear to play roles in plant defense mechanisms, they were evaluated for their effects on economically important plant pathogenic fungi of the genera Colletotrichum, Botrytis, Fusarium, and Phomopsis. We further investigated these peanut phytoalexins, together with some related natural and synthetic stilbenoids (a total of 24 compounds) in a panel of bioassays to determine their anti-inflammatory, cytotoxic, and antioxidant activities in mammalian cells. Several of these compounds were also evaluated as mammalian opioid receptor competitive antagonists. Assays for adult mosquito and larvae toxicity were also performed. The results of these studies reveal that peanut stilbenoids, as well as related natural and synthetic stilbene derivatives, display a diverse range of biological activities.     

Localized production of phytoalexins by peanut (Arachis hypogaea) kernels in response to invasion by Aspergillus species

Peanuts respond to fungal infection by synthesizing phytoalexins, most of which are antibiotic stilbenes. The mechanism and dynamics of phytoalexin formation in the peanut have not been studied. One of the most popular peanut cultivars in the southeastern United States, Georgia Green, was investigated for its ability to produce phytoalexins in response to infection by soil fungal strains. The experimental design allowed for study of phytoalexin production in peanut kernels layer-by-layer. The layers were dissected from different depths of the kernel starting from the infected area down to healthier tissues. Six peanut phytoalexins, trans-resveratrol, trans-arachidin-1, trans-arachidin-2, trans-arachidin-3, trans-3-isopentadienyl-4,3',5'-trihydroxystilbene, and SB-1, were detected in the kernel slices and quantitated. All of the fungal strains triggered phytoalexin production; however, the composition of phytoalexins varied significantly by layer. After incubation for 24 h, tissues remote from the infected area produced almost exclusively trans-resveratrol, whereas closer to the infected area tissues synthesized all six phytoalexins. In all of the experiments, after 48 h of fungal growth, deeper layers produced all tested phytoalexins. There was a significant difference in phytoalexin production elicited by some fungal isolates. No association was observed between phytoalexin production and toxigenic potential of fungal strains that elicited the production in mature peanut kernels.     

trans-resveratrol content in commercial peanuts and peanut products

A modified high-performance liquid chromatographic (HPLC) method for determination of trans-resveratrol (resveratrol) in peanuts and peanut products has been developed. Resveratrol was extracted with acetonitrile-water (90/10, v/v) by blending with diatomaceous earth at high speed followed by purification of an aliquot of the extract on a minicolumn packed with Al(2)O(3)-ODS (C(18)) mixture. The column was eluted with acetonitrile-water (90/10, v/v), eluate was evaporated under nitrogen, and residue was dissolved in HPLC mobile phase. Resveratrol in an aliquot of purified extract was quantitated by HPLC on silica gel with n-hexane-2-propanol-water-acetonitrile-acetic acid (1050/270/17/5/1, v/v) as a mobile phase. The recovery of resveratrol added to diatomaceous earth at 0.05 microg/g was 98.95 +/- 17.79%; the recovery of the standard added to fresh peanuts (with 0.070 microg/g natural level of resveratrol) at 0.50, 5.00, and 10.00 microg/g was 117.23 +/- 8.87, 100.10 +/- 2.49, and 100.45 +/- 1.51%, respectively. The quantitation limit of resveratrol in fresh peanuts was about 0. 01 microg/g. Roasted peanuts had the lowest content of resveratrol of 0.055 +/- 0.023 microg/g (n = 21), while in peanut butter its concentration was significantly higher, 0.324 +/- 0.129 microg/g (n = 46), and boiled peanuts had the highest level of 5.138 +/- 2.849 microg/g (n = 12). Resveratrol content in commercial peanut products was similar to the resveratrol content of the raw peanut fractions routinely used for making them.     

Peanut roots as a source of resveratrol

A potent antioxidant, resveratrol (3,4',5-trihydroxystilbene), was extracted using 80% methanol from peanut roots (Arachis hypogaea L.), isolated with a solid-phase extraction column, purified by a semipreparative HPLC, and identified with 1H NMR and MS. The highest and lowest resveratrol contents in the peanut roots of 2000 fall and 2001 spring crops were 1.330 and 0.130 mg/g and 0.063 and 0.015 mg/g, respectively. When the dehydrated peanut root powders of spring and fall crops were combined and cooked with pork-fat patties (1%, w/w) and the separated oils were stored at 60 degrees C for conjugated diene hydroperoxide (CDHP) determination, CDHP contents of the control oils increased after 3 days of storage, whereas the contents in the peanut root-treated oils of spring and fall crops did not increase after 9 and 15 days of storage, respectively. It is of merit to find that peanut roots, usually left in the field as agricultural waste, contain resveratrol and bear potent antioxidative activity.     

Determination of the phytoalexin resveratrol (3,5,4'-trihydroxystilbene) in peanuts and pistachios by high-performance liquid chromatographic diode array (HPLC-DAD) and gas chromatography-mass spectrometry (GC-MS)

The phytoalexin resveratrol (3,5,4'-trihydroxystilbene) in edible peanut (Arachis hypogaea L.) and pistachio (Pistacia vera L.) varieties grown in Turkey was analyzed by high-performance liquid chromatographic diode array and gas chromatography-mass spectrometric detection. trans-Resveratrol in six peanut varieties, five pistachio varieties, and four market samples ranged between 0.03 and 1.92 microg/g. The Cerezlik 5025 peanut (1.92 +/- 0.01 microg/g) and Ohadi pistachio genotype (1.67 +/- 0.01 microg/g) had significantly higher trans-resveratrol contents. Peanuts contained 0.03-1.92 microg/g (av = 0.84 microg/g) of trans-resveratrol, whereas pistachio contained 0.09-1.67 microg/g (av = 1.15 microg/g). With exposure to UV light for 1 min, trans-resveratrol concentrations of samples ranged from 0.02 to 1.47 microg/g and those of cis-resveratrol from 0.008 to 0.32 microg/g. The occurrence of resveratrol in peanut and pistachio was confirmed by total ion chromatograms (TIC) of bis[trimethylsilyl]trifluoroacetamide derivatives of resveratrol isomers and comparison of the mass spectral fragmentation data with those of a resveratrol standard. Formation of the cis-isomer in pistachios was higher than in peanuts.     

Cancer chemopreventive and antioxidant activities of pterostilbene,a naturally occurring analogue of resveratrol

Pterostilbene, a natural methoxylated analogue of resveratrol, was evaluated for antioxidative potential. The peroxyl-radical scavenging activity of pterostilbene was the same as that of resveratrol, having total reactive antioxidant potentials of 237 +/- 58 and 253 +/- 53 microM, respectively. Both compounds were found to be more effective than Trolox as free radical scavengers. Using a plant system, pterostilbene also was shown to be as effective as resveratrol in inhibiting electrolyte leakage caused by herbicide-induced oxidative damage, and both compounds had the same activity as alpha-tocopherol. Pterostilbene showed moderate inhibition (IC50 = 19.8 microM) of cyclooxygenase (COX)-1, and was weakly active (IC50 = 83.9 microM) against COX-2, whereas resveratrol strongly inhibited both isoforms of the enzyme with IC50 values of approximately 1 microM. Using a mouse mammary organ culture model, carcinogen-induced preneoplastic lesions were, similarly to resveratrol, significantly inhibited by pterostilbene (ED50 = 4.8 microM), suggesting antioxidant activity plays an important role in this process.     

Crocetin, dimethylcrocetin, and safranal bind human serum albumin: stability and antioxidative properties

Crocetin (CRT) and dimethylcrocetin (DMCRT) are derived from crocins which are found in the stigmas of saffron (Crocus sativus L.), while safranal is the main component of saffron's essential oil. The aim of the present study was to examine their interaction with human serum albumin in aqueous solution at physiological conditions using constant protein concentration and various ligand contents. FT-IR and UV-visible spectroscopic methods were used to determine the ligands' binding mode, the binding constant, and the effects of ligand complexation on protein secondary structure. Structural analysis showed that crocetin, dimethylcrocetin, and safranal bind nonspecifically (H-bonding) via protein polar groups with binding constants of Kcrt =2.05 (+/-0.30) x 103 M-1, Kdmcrt = 9.60 (+/-0.35) x 104 M-1, and Ksaf = 2.11 (+/-0.35) x 103 M-1. The protein secondary structure showed no major alterations at low ligand concentrations (1 microM), whereas at higher content (1 mM), decrease of alpha-helix from 55% (free HSA) to 43-45% and increase of beta-sheet from 17% (free HSA) to 18-22% and random coil 7% (free HSA) to 10-14% occurred in the ligand-HSA complexes. The results point to a partial unfolding of protein secondary structure at high ligand content. The antioxidant activity of CRT, DMCRT, and safranal was also tested by the DPPH* antioxidant activity assay, and their IC50 values were compared to that of well-known antioxidants such as Trolox and butylated hydroxy toluene (BHT). The IC50 values of CRT and safranal were 17.8 +/- 1 microg/mL and 95 +/- 1 microg/mL, respectively, while the inhibition of DMCRT reached a point of 38.8%, which corresponds to a concentration of 40 microg/mL, and then started to decrease. The IC50 values of Trolox and BHT were 5.2 +/- 1 microg/mL and 5.3 +/- 1 microg/mL, respectively.     

Interrelationship of phytoalexin production and disease resistance in selected peanut genotypes

In peanuts, a mechanism of resistance to fungal infection is reportedly due to the synthesis of stilbene phytoalexins, which are antibiotic, low molecular weight metabolites. The phytoalexin-associated response of different peanut genotypes to exogenous invasion in the field has not been investigated and may be useful for breeding resistant peanut cultivars. Five peanut genotypes, Georgia Green, Tifton 8, C-99R, GK-7 High Oleic, and MARC I, which differ in resistance to major peanut diseases, were investigated for their ability to produce phytoalexins under field conditions in South Georgia in 2001 and 2002. Five known peanut phytoalexins, trans-resveratrol, trans-arachidin-1, trans-arachidin-2, trans-arachidin-3, and trans-3'-isopentadienyl-3,5,4'-trihydroxystilbene, were quantitated. The phytoalexins were measured in peanuts of different pod maturity (yellow, orange, brown, and black) with or without insect pod damage (externally scarified or penetrated). Kernels from insect-damaged pods of C-99R and Tifton 8 genotypes had significantly higher concentrations of phytoalexins than other genotypes. The same genotypes were the most resistant to tomato spotted wilt virus and late leaf spot, while MARC I, which is highly susceptible to these diseases, produced very low concentrations of phytoalexins. However, there was no significant difference in phytoalexin production by undamaged peanut pods of all tested genotypes. trans-Arachidin-3 and trans-resveratrol were the major phytoalexins produced by insect-damaged peanuts. In damaged seeds, the concentrations of trans-3'-isopentadienyl-3,5,4'-trihydroxystilbene were significantly higher in Tifton 8 as compared to other genotypes. There was an association between total phytoalexin production and published genotype resistance to major peanut diseases. Stilbene phytoalexins may be considered potential chemical markers in breeding programs for disease-resistant peanuts.     

Resveratrol oligomers isolated from Carex species inhibit growth of human colon tumorigenic cells mediated by cell cycle arrest

Research has shown that members of the Carex genus produce biologically active stilbenoids including resveratrol oligomers. This is of great interest to the nutraceutical industry given that resveratrol, a constituent of grape and red wine, has attracted immense research attention due to its potential human health benefits. In the current study, five resveratrol oligomers (isolated from Carex folliculata and Carex gynandra ), along with resveratrol, were evaluated for antiproliferative effects against human colon cancer (HCT-116, HT-29, Caco-2) and normal human colon (CCD-18Co) cells. The resveratrol oligomers included one dimer, two trimers, and two tetramers: pallidol (1); α-viniferin (2) and trans-miyabenol C (3); and kobophenols A (4) and B (5), respectively. Although not cytotoxic, the resveratrol oligomers (1-5), as well as resveratrol, inhibited growth of the human colon cancer cells. Among the six stilbenoids, α-viniferin (2) was most active against the colon cancer cells with IC(50) values of 6-32 μM (>2-fold compared to normal colon cells). Moreover, α-viniferin (at 20 μM) did not induce apoptosis but arrested cell cycle (in the S-phase) for the colon cancer but not the normal colon cells. This study adds to the growing body of knowledge supporting the anticancer effects of resveratrol and its oligomers. Furthermore, Carex species should be investigated for their nutraceutical potential given that they produce biologically active stilbenoids such as α-viniferin.     

Occurrence of resveratrol in edible peanuts

Resveratrol has been associated with reduced cardiovascular disease and reduced cancer risk. This phytoalexin has been reported in a number of plant species, including grapes, and may be one of the compounds responsible for the health benefits of red wine. Analytical methods for measuring resveratrol in wine and peanuts were adapted to isolate, identify, and quantify resveratrol in several cultivars of peanuts. Aqueous ethanol (80% v/v) extracts from peanuts without seed coats were purified over alumina/silica gel columns and analyzed by reversed phase HPLC using a C-18 column. Peanuts from each market type, Virginia, runner, and Spanish, produced in four different locations contained from 0.03 to 0.14 microg of resveratrol/g. Seed coats from runner and Virginia types contained approximately 0.65 microg/g of seed coat, which is equivalent to <0.04 microg/seed. Quantitative analysis of 15 cultivars representing 3 peanut market types, which had been cold stored for up to 3 years, indicated a range of 0.02-1.79 microg/g of peanut compared to 0.6-8.0 microg/mL in red wines.     

Cyclooxygenase inhibitory and antioxidant compounds from crabapple fruits

Crabapple trees belong to the Malus genus (Rosaceae) and bear fruits that are sparingly consumed and used in the preparation of fruit beverages. Cyclooxygenase (COX) enzyme inhibitory and antioxidant bioassay-guided fractionation of the aqueous and methanol extracts of Malus x kornicensis and Malus x Indian Summer yielded (+)-catechin (1), (-)-epicatechin (2), cyanidin-3-O-beta-galactopyranoside (3), and amygdalin (4). Pure compounds 1-4 were obtained by HPLC, identified by LC-ES/MS, CD, and NMR spectroscopic methods and evaluated for their COX enzyme inhibitory and antioxidant activities. In COX-1 and -2 enzyme inhibitory assays, compounds 1-3 (all at 80 microM) showed activities of 20.4, 46.3%; 57.6, 47.9%; and 8.2, 13.7%, respectively, compared to naproxen (54.3, 41.3%; 10 microM), ibuprofen (47.5, 39.8%; 10 microM), Celebrex (46.2, 66.3%; 1.67 ppm), and Vioxx (23.8, 88.1%, 1.67 ppm). In the antioxidant assay, the catechins (1-2) and anthocyanin (3) (all at 40 microM) showed activities of 61.3, 62.5, and 60.1%, respectively. The synthetic antioxidants, tert-butylhydroquinone (TBHQ), butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), and vitamin E (all tested at 10 microM), gave 75.2, 80.1, 70.0, and 10.2% activities, respectively. The cyanogenic glycoside, amygdalin (4), and its hydrolysis products, mandelonitrile (5) and benzaldehyde (6), were not active in the antioxidant or COX enzyme inhibitory assays at 80 microM concentrations.     

Sequestering ability of butylated hydroxytoluene,propyl gallate,resveratrol, and vitamins C and E against ABTS,DPPH, and hydroxyl free radicals in chemical and biological systems

The antioxidant capacity of butylated hydroxytoluene (BHT; 2,6-di-tert-butyl-p-cresol), propyl gallate (3,4,5-trihydroxybenzoic acid n-propyl ester), resveratrol (trans-3,4',5-trihydroxystilbene), and vitamins C (l-ascorbic acid) and E [(+)-alpha-tocopherol] was studied in chemical and biological systems. The chemical assays evaluated the capacity of these antioxidants to sequester 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS.) and 1,1 diphenyl-2-picrylhydrazyl (DPPH.). A new colorimetric method to determine hydroxyl radical scavenging is also described. The biological tests use the eucaryotic cells of Saccharomyces cerevisiae treated with the antioxidants in the presence of the stressing agents apomorphine, hydrogen peroxide, and paraquat dichloride (methylviologen; 1,1'-dimethyl-4,4'-bipyridinium dichloride). The results in chemical systems showed that all of the antioxidants were able to significantly inhibit the oxidation of beta-carotene by hydroxyl free radicals. The assays in yeast showed that the antioxidant activity of the tested compounds depended on the stressing agent used and the mechanism of action of the antioxidant.     

Germination of peanut kernels to enhance resveratrol biosynthesis and prepare sprouts as a functional vegetable

Bioactive benefits of resveratrol in the diets have attracted extensive interests of the public. Peanut is one of the potent natural sources of resveratrol. In this study, germination of peanut kernels to enhance resveratrol biosynthesis and preparation of sprouts as a functional vegetable was conducted. When the rehydrated kernels of three peanut cultivars were germinated at 25 degrees C and relative humidity 95% in dark for 9 days, resveratrol contents increased significantly from the range of 2.3 to 4.5 microg/g up to the range of 11.7 to 25.7 mug/g depending upon peanut cultivar. In comparison with the sprout components, resveratrol contents were highest in the cotyledons, slightly lower in the roots, and not detected in the stems. When the sprouts were heated in boiling water for 2 min, resveratrol contents varied in a limited range. Methanol extracts of the freeze-dried sprouts exhibited potent 1,1-diphenyl-2-picryl-hydrazyl scavenging activity and antioxidative potency against linoleic acid oxidation. These activities increased with an increase of germination time. After 9 days of germination, total free amino acid, sucrose, and glucose contents increased significantly while crude protein contents decreased and the large sodium dodecyl sulfate polyacrylamide gel electrophoresis protein molecules of the kernels were extensively degraded. From a practical viewpoint, it is of potency to prepare peanut sprouts as a functional vegetable.     

Detection and quantitation of resveratrol in tomato fruit (Lycopersicon esculentum Mill.)

Resveratrol is a stilbene phytoalexin well-known for its presence in grape, wine, and peanut. As a result of its antioxidant and chemopreventative properties, it has gained much attention as a functional food ingredient. A gas chromatography-mass spectrometry method for the detection of resveratrol, its 3-glucopyranoside piceid, and the cis isomers of both compounds has been developed and used to quantitate the levels of these compounds in the skin of commercially available tomato fruit (Lycopersicon esculentum Mill.). The resveratrol concentration remains relatively stable during fruit maturation, reaching a maximum concentration in the skin of 18.4 +/- 1.6 microg/g dry weight at 4 weeks postbreaker. No stilbenes were detected in the flesh of tomato fruit.     

Comparison of ozone and UV-C treatments on the postharvest stilbenoid monomer, dimer, and trimer induction in var. 'Superior' white table grapes

Postharvest treatment of seedless white table grapes (var. 'Superior') with different gas ozone concentrations (3.88 and 1.67 g/h) for 1, 3, and 5 h induced an increase in stilbenoid biosynthesis [trans-resveratrol, piceatannol, and viniferinas (resveratrol dehydrodimers and dehydrotrimers)] during storage at 22 degrees C and 95% relative humidity. The maximal resveratrol concentration was reached after 2 days of storage, and this amount was similar to that induced by optimized UV-C treatments (1 min, 510 W, 40 cm). Although similar resveratrol concentrations accumulated in grapes after both UV-C and O3 treatments (maximum ozone production and time), the ozone treatment was more efficient in inducing viniferins accumulation in grape berries. A sequence in the biosynthesis of stilbenoids was observed, starting with the resveratrol monomer, continuing with the resveratrol dehydrodimers epsilon-viniferin and delta-viniferin, and ending with four different resveratrol dehydrotrimers. These trimers were different from alpha-viniferin, a trimer previously reported to be induced in grapes after biotic and abiotic stresses. Two alpha-viniferin isomers were also detected in the ozone-treated grapes, although at very low concentrations that prevented their quantification.     

Catalytic inhibition of human DNA topoisomerase II by interactions of grape cell culture polyphenols

Previously, we isolated mixed polyphenolic fractions on a toyopearl matrix (TP-2 to TP-6) from grape cell cultures that were highly potent catalytic inhibitors in a human DNA topoisomerase II assay for cancer chemoprevention. The objectives of this study were to evaluate the potency of, and potential interactions between, individual fractions and some of the purified bioactive polyphenols that comprise these fractions on human DNA topoisomerase II catalytic activity. Treatments that combined anthocyanin-rich fractions (TP-2; 0.5 or 2.0 microg of dried material/mL), fractions containing catechins, procyanidin dimers, and flavanones (TP-4; 0.25 microg of dried material/mL), and/or fractions enriched with procyanidin oligomers and polymers (TP-6; 0.15 or 0.5 microg of dried material/mL) showed additive effects toward catalytic inhibition of the enzyme. Epicatechin gallate (IC50 = 0.029 microM), myricetin (0.39 microM), procyanidin B2 (PB2, 4.5 microM), and resveratrol (65.7 microM), constituents of the most bioactive mixed fraction from grape cell culture (TP-4), each individually provided potent catalytic inhibition of topoisomerase II. In addition, potentiating interactions between the PB2 and the other polyphenolic constituents mentioned above and between myricetin and resveratrol were clearly demonstrated. A synergistic interaction between myricetin and resveratrol was also confirmed with isobolographic analysis at a molar ratio of 1:70.     

Characterization of resveratrol content in ten wild species of section Arachis, genus Arachis

Peanut is one of the few plants that synthesizes resveratrol, a phenolic compound of the stilbene class, which has been associated with reduced risk of developing chronic diseases, such as cancer, cardiovascular diseases, skin diseases, pulmonary diseases, diabetes and neurological diseases. Resveratrol was detected in different parts of the peanut plant, including roots, leaves, seeds and their derivatives. The wild species of the Arachis section are also strong candidates to synthesize resveratrol because they are phylogenetically closely related to cultivated peanut. Our objective was to characterize the resveratrol content in ten wild species of Arachis with three different genomes (A, B and K). The plant material was composed of leaves of the ten species treated (test) and not treated (control) with ultraviolet (UV) radiation. The test and control samples were extracted and the identification and quantification of resveratrol was performed using high performance liquid chromatography (HPLC). All species studied synthesized resveratrol and the concentrations ranged from 299.5 μg/g in A. kempff-mercadoi to 819.9 μg/g in A. cardenasii. DPPH antioxidant activity varied between 18.7 % for A. duranensis and 48.2 % in A. simpsonii. The results showed that wild Arachis species are a potential source of alleles for improvement of cultivated peanut, with the aim of achieving higher resveratrol content in leaves.     

Antioxidant evaluation in dessert spices compared with common food additives. Influence of irradiation procedure

The antioxidant properties of seven dessert spices (anise, cinnamon, ginger, licorice, mint, nutmeg, and vanilla) were compared with those of the common food antioxidants butylated hydroxyanisole (BHA) (E-320), butylated hydroxytoluene (BHT) (E-321), and propyl gallate (E-310). The influence of irradiation process on antioxidant activity was also evaluated. Mint and cinnamon exhibited a higher percentage of inhibition of oxidation than the other spices analyzed and the food antioxidants, as tested by the lipid peroxidation assay (LOO*). Nutmeg, anise, and licorice showed the strongest protection in the deoxyribose assay (OH*). Vanilla exhibited the highest antioxidant activity in the peroxidase-based assay (H2O2). Nutmeg, propyl gallate, ginger, and licorice improved the stability of oils (sunflower, corn, and olive) and fats (butter and margarine) against oxidation (110 degrees C Rancimat). Cinnamon was a better superoxide radical scavenger than the other analyzed spices and additives. When the Trolox equivalent antioxidant capacity (TEAC) assay was used to provide a ranking order of antioxidant activity, the result in decreasing order of antioxidant capacity was cinnamon approximately equal to propyl gallate > mint > anise > BHA > licorice approximately equal to vanilla > ginger > nutmeg > BHT. Irradiated samples did not show significant differences (p < 0.05) in the antioxidant activity with respect to the non-irradiated samples (1, 3, 5, and 10 kGy) in the assays used.     

Potential food additives from Carex distachya roots: identification and in vitro antioxidant properties

In the present study, the chemical composition and antioxidant properties of root methanol extract of Carex distachya Desf. (Cyperaceae) were assessed to use this plant as sources of food additives and nutraceuticals. The IC50 of the extract (4.2 microg/mL), derived from the DPPH radical scavenging capacity assay, was similar to those of ascorbic acid, alpha-tocopherol, and BHT. These results revealed a strong antioxidant activity because of the presence of an extraordinary quantity of bioactive phytochemicals. The phytochemical study of the root extract led to the isolation and identification of new and known polyphenols, most of them common constituents of plant foods. A total of 16 polyphenols, identified on the basis of spectroscopic data as 7 lignans, 4 phenylethanoids, 3 resveratrol derivatives, a monolignol, and a secoiridoid glucoside, were isolated. The tentative structural elucidation of the new metabolites 5'-O-beta-D-glucopyranosyloxy-3,3'-dimethoxy-7,9'-epoxylignan-4,8',9-triol and 3,5-bis-O-beta-D-glucopyranosyloxy-3'-methoxy-trans-stilben-4'-ol have been performed by a combined approach using ESI/TQ/MS techniques and 1D and 2D NMR experiments. All of the compounds have been tested for their antioxidant activity using six different antioxidant and radical scavenging tests. Interestingly, the extract contained high quantities of polyphenols, most of them reported as constituents of edible plants, such as grape and olive, suggesting that the methanol root extract of this plant could be used as a source of natural antioxidants useful as potential food additives.