Epilobium angustifolium L. Extracts as Valuable Ingredients in Cosmetic and Dermatological Products

Epilobium angustifolium L. is a popular and well-known medicinal plant. In this study, an attempt to evaluate the possibility of using this plant in preparations for the care and treatment of skin diseases was made. The antioxidant, antiaging and anti-inflammatory properties of ethanolic extracts from Epilobium angustifolium (FEE) were assessed. Qualitative and quantitative evaluation of extracts chemically composition was performed by gas chromatography with mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC). The total polyphenol content (TPC) of biologically active compounds, such as the total content of polyphenols (TPC), flavonoids (TFC), and assimilation pigments, as well as selected phenolic acids, was assessed. FEE was evaluated for their anti-inflammatory and antiaging properties, achieving 68% inhibition of lipoxygenase activity, 60% of collagenase and 49% of elastase. FEE also showed high antioxidant activity, reaching to 87% of free radical scavenging using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 59% using 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS). Additionally, in vitro penetration studies were performed using two vehicles, i.e., a hydrogel and an emulsion containing FEE. These studies showed that the active ingredients contained in FEE penetrate through human skin and accumulate in it. The obtained results indicate that E. angustifolium may be an interesting plant material to be applied as a component of cosmetic and dermatological preparations with antiaging and anti-inflammatory properties.     

Comparative Study of the Chemical Compositions and Antioxidant Activities of Fresh Juices from Romanian Cucurbitaceae Varieties

Cucurbitaceae is a family of health-promoting plants due to their compounds with beneficial effects. The aim of this study was to analyze, for the first time, the chemical composition, the antioxidant activity and the metal chelating properties of fruit juices obtained from four different species of the Cucurbitaceae family cultivated in Romania, namely Momordica charantia, Cucumis metuliferus, Benincasa hispida and Trichosanthes cucumerina. The samples of juice were analyzed by high-performance liquid chromatography (HPLC) and all the four species displayed high levels of the two triterpenes, oleanolic and ursolic acids, and also in phenolic compounds, including catechin, (−)-epicatechin and gallic acid. The juices demonstrated significant antioxidant activity against the free radical 2,2-diphenyl-1-picrylhydrazyl (ranging from 20 to 95%,), a good iron binding ability (ranging from 7.45 ± 0.28% to 86.95 ± 0.97%) and also promising antioxidant potential against the ABTS radical (ranging from 4.97 to 32.60 μETx/mL juice). Our findings raise interesting questions for further research on Cucurbitaceae fruit juices and, consequently, their very good antioxidant potential suggests these fruits should be further explored for their protective effect against oxidative damage. This is the first time the chemical composition and antioxidant activities of fruit juices from these four Romanian Cucurbitaceae varieties have been investigated.     

Antioxidant potential and authenticity of some commercial fruit juices studied by EPR and IRMS

Antioxidant status of foods, plant, or fruit products is generally characterized by means of spectroscopic methods. Methods like HPLC, UV-VIS, or MS spectroscopy are used to understand the chemical and physical properties of different samples, and also EPR spectroscopy seems to be a valuable tool to characterize antioxidant activity of juice beverages. In this technique, certain antioxidants present in fruit juices interact with free radicals interrupting the chain reaction that can possibly damage essential molecules. Recording the EPR signal decay caused by the reaction with a natural or artificial reducing agent, it is possible to draw conclusions about the antioxidant capability of materials. IRMS is a powerful tool to distinguish between an authentic fruit juice and a juice obtained by concentrate dilution. This technique allows also the detection of commercial C₄ cane or corn derived sugar syrups in C₃ fruit juices. In the present study, four commercial fruit juices were investigated using stable isotope measurements (oxygen, hydrogen, and carbon) and EPR measurements in order to check the correct labeling in the Romanian markets and to compare antioxidant activity of the studied juices and the reference. It was proven that the number of paramagnetic species decreases in time with different reaction rates and this was correlated with the antioxidant activity of the studied juices. Stable isotope ratio measurements have demonstrated that the fruit juices studied were reconstructed from concentrates with tap water, according to their label.     

Physicochemical,Antioxidant, Microstructural Properties and Bioaccessibility of Dark Chocolate with Plant Extracts

In this study, dark chocolates (DCh) containing zinc lactate (ZnL) were enriched with extracts from elderberries (EFrE), elderflowers (EFlE), and chokeberries (ChFrE) to improve their functional properties. Both dried plant extracts and chocolates were analyzed for antioxidant capacity (AC) using four different analytical methods: 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), cupric ion-reducing antioxidant capacity (CUPRAC), and ferric-reducing antioxidant power (FRAP), while total phenolic content (TPC) was determined by Folin–Ciocalteu (F–C) assay. An increase in antioxidant properties of fortified chocolates was found, and the bioaccessibility of their antioxidants was evaluated. The highest AC and TPC were found in ChFrE and chocolate with chokeberries (DCh + ChFrE) before and after simulated in vitro digestion. Bioaccessibility studies indicated that during the simulated digestion the AC of all chocolates reduced significantly, whereas insignificant differences in TPC results were observed between chemical and physiological extracts. Moreover, the influence of plant extracts on physicochemical parameters such as moisture content (MC), fat content (FC), and viscosity of chocolates was estimated. Furthermore, scanning electron microscopy with dispersive energy spectroscopy (SEM-EDS) was used to analyze surface properties and differences in the chemical composition of chocolates without and with additives.     

Antioxidant, anti-lipoxygenase and cytotoxic activity of Leptadenia pyrotechnica (Forssk.) decne polyphenolic constituents

Leptadenia pyrotechnica Forssk is a traditional medicinal herb used for treatment of inflammatory diseases and cancer. In this research, the aqueous ethanolic crude extract of Leptadenia pyrotechnica aerial parts, along with its ethyl acetate, n-butanol and water partitioning fractions were evaluated for their antioxidant capacity, polyphenolic content, anti-inflammatory and anti-cancer properties. The total antioxidant capacity was estimated by the FRAP, DPPH, ABTS and β-carotene bleaching assays.The ethyl acetate fraction exhibited the highest polyphenolic content (252.27 mg gallic acid/g) and the best antioxidant activity (1.2, 0.57, 0.45 mmol ascorbic acid equivalent/g in the FRAP, ABTS and DPPH assays, respectively). Furthermore, the same extract showed appreciable anti-inflammatory via lipoxygenase (LOX) inhibitory activity (IC?? = 1.41 μg/mL). Moreover, the ethyl acetate fraction also showed the strongest cytotoxic effect (IC?? = 43.16 μg/mL) against MCF-7 human breast cancer cell line. These results suggest that this plant may be considered an interesting source of compounds with antioxidant, anti-inflammatory and anti-cancer properties for therapeutic, nutraceutical and functional food applications.     

Studies on the potential antioxidant properties of Senecio stabianus Lacaita (Asteraceae) and its inhibitory activity against carbohydrate-hydrolysing enzymes

This study showed for the first time the antioxidant and hypoglycaemic properties of the methanol, n-hexane and ethyl acetate extracts from Senecio stabianus Lacaita, a plant that belongs to the Asteraceae family. The antioxidant activities were carried out using two different in vitro assays, namely 2,2-diphenyl-1-picrylhydrazyl (DPPH) test and 2,2'-azinobis(3-ethylbenzthiazoline-6-sulphonate) (ABTS) test. The ethyl acetate extract showed the highest activity with inhibitory concentration 50% (IC(50)) values of 35.5 and 32.7?μg?mL(-1) on DPPH test and ABTS test, respectively. This activity may be related to a good total phenol and flavonoid content. All extracts were also tested for their potential inhibitory activity of α-amylase and α-glucosidase digestive enzymes. The n-hexane extract exhibited the highest α-amylase inhibition with an IC(50) value of 0.21?mg?mL(-1). Through bioassay-guided fractionation processes seven fractions (A-G) were obtained and tested. Based on the phytochemical analysis, the activity of n-hexane extract may be related to the presence of monoterpenes and sesquiterpenes.     

Synthesis and Evaluation of Antioxidant Properties of 2-Substituted Quinazolin-4(3H)-ones

Quinazolinones represent an important scaffold in medicinal chemistry with diverse biological activities. Here, two series of 2-substituted quinazolin-4(3H)-ones were synthesized and evaluated for their antioxidant properties using three different methods, namely DPPH, ABTS and TEAC_CUPRAC, to obtain key information about the structure–antioxidant activity relationships of a diverse set of substituents at position 2 of the main quinazolinone scaffold. Regarding the antioxidant activity, ABTS and TEAC_CUPRAC assays were more sensitive and gave more reliable results than the DPPH assay. To obtain antioxidant activity of 2-phenylquinazolin-4(3H)-one, the presence of at least one hydroxyl group in addition to the methoxy substituent or the second hydroxyl on the phenyl ring in the ortho or para positions is required. An additional ethylene linker between quinazolinone ring and phenolic substituent, present in the second series (compounds 25a and 25b), leads to increased antioxidant activity. Furthermore, in addition to antioxidant activity, the derivatives with two hydroxyl groups in the ortho position on the phenyl ring exhibited metal-chelating properties. Our study represents a successful use of three different antioxidant activity evaluation methods to define 2-(2,3-dihydroxyphenyl)quinazolin-4(3H)-one 21e as a potent antioxidant with promising metal-chelating properties.     

Antioxidant and Antiglycation Effects of Cistus × incanus Water Infusion,Its Phenolic Components,and Respective Metabolites

Reactive oxygen and carbonyl species promote oxidative and carbonyl stress, and the development of diabetes, metabolic syndrome, cardiovascular diseases, and others. The traditional herb Cistus × incanus is known for its antioxidant properties; therefore, the current study aimed to assess how the chemical composition of a C. incanus water infusion corresponds with its antioxidative and antiglycative effects in vitro. The composition of infusions prepared from commercial products was analyzed with UHPLC-ESI-qTOF-MS. Total phenolics, flavonoids, and non-flavonoid polyphenols were determined. Antioxidant activity of infusions and selected polyphenols was investigated using DPPH, ABTS, and FRAP. Fluorometric measurements and methylglyoxal capture were performed to investigate the antiglycation activity. PCA and PLS-DA models were applied to explore the correlation between chemical and antioxidant results. The principal flavonoids in C. incanus were flavonols. In vitro tests revealed that a stronger antioxidant effect was demonstrated by plant material from Turkey rich in flavonoids, followed by Albania and Greece. Flavonols and ellagic acid displayed stronger antiradical and reducing power than EA-derived urolithins. Hyperoside was the most potent inhibitor of glycation. The results indicate that flavonoids are primarily responsible for rock rose antioxidant and antiglycation properties. PLS-DA modeling can be used to identify the origin of plant material with sensitivity and specificity exceeding 86%.     

Actinidia arguta Leaf as a Donor of Potentially Healthful Bioactive Compounds: Implications of Cultivar,Time of Sampling and Soil N Level

The aim of this study was to assess the enzymatic and non-enzymatic antioxidant status of kiwiberry (Actinidia arguta) leaf under different N regimes tested three times in field conditions during the 2015 growing season in two cultivars (‘Weiki’ and ‘Geneva’). Leaf total antioxidant capacity using ABTS, DPPH and FRAP tests was evaluated in the years 2015 to 2017, which experienced different weather conditions. Both cultivars exhibited a significant fall in leaf L-ascorbic acid (L-AA) and reduced glutathione (GSH) as well as global content of these compounds during the growing season, while total phenolic contents slightly (‘Weiki’) or significantly (‘Geneva’) increased. There was a large fluctuation in antioxidative enzyme activity during the season. The correlation between individual antioxidants and trolox equivalent antioxidant capacity (TEAC) depended on the plant development phase. The study revealed two peaks of an increase in TEAC at the start and end of the growing season. Leaf L-AA, global phenolics, APX, CAT and TEAC depended on the N level, but thiol compounds were not affected. Over the three years, TEAC decreased as soil N fertility increased, and the strength of the N effect was year dependent. The relationship between leaf N content and ABTS and FRAP tests was highly negative. The antioxidant properties of kiwiberry leaves were found to be closely related to the plant development phase and affected by soil N fertility.     

Polyphenol content and antioxidant activity of fourteen wild edible fruits from Burkina Faso

A total of fourteen (14) species of wild edible fruits from Burkina Faso were analyzed for their phenolic and flavonoid contents, and their antioxidant activities using the DPPH, FRAP and ABTS methods. The data obtained show that the total phenolic and total flavonoid levels were significantly higher in the acetone than in the methanol extracts.Detarium microcarpum fruit had the highest phenolic and the highest flavonoid content,followed by that of Adansonia digitata, Ziziphus mauritiana, Ximenia americana and Lannea microcarpa. Significant amounts of total phenolics were also detected in the other fruit species in the following order of decreasing levels: Tamarindus indica > Sclerocaryabirrea > Dialium guineense > Gardenia erubescens > Diospyros mespiliformis > Parkiabiglobosa > Ficus sycomorus > Vitellaria paradoxa. Detarium microcarpum fruit also showed the highest antioxidant activity using the three antioxidant assays. Fruits with high antioxidant activities were also found to possess high phenolic and flavonoid contents. There was a strong correlation between total phenolic and flavonoid levels and antioxidant activities.     

Global chemical composition and antioxidant and anti-tuberculosis activities of various extracts of Globularia alypum L. (Globulariaceae) leaves

In this work, an evaluation of the biological activities of Globularia alypum L. extracts and their global chemical composition was realized. Extracts from G. alypum were obtained by two extraction methods. The composition of polyphenols (8.5-139.95 g gallic acid equivalent/Kg of dry mass), tannins (1.39-18.65 g catechin equivalent/Kg of dry mass), anthocyanins (8.17-70.69 mg cyanidin equivalent/Kg of dry mass) and flavonoids (0.31-19.28 g quercetin equivalent/Kg of dry mass) was evaluated. The samples were subjected to a screening for their antioxidant activities using the DPPH* and ABTS*+ assays. For the first time, the anti-tuberculosis activity (H(37)Rv) for G. alypum was tested against Mycobacterium tuberculosis. The strongest antioxidant activity was obtained for the methanol extract (IC(50 ) = 15.58 ± 0.168 mg/L) and the best anti-tuberculosis activity was obtained for the petroleum ether extract (IC(50 )= 77 mg/L). We have found a positive correlation between the total phenolics content and the antioxidant activity R2 = 0.88 (DPPH*) and R2 = 0.97 (ABTS*+). We have found also a positive correlation between the flavonoid content and the antioxidant activity R2 = 0.91 (DPPH*) and R2 = 0.91 (ABTS*+).     

Antioxidant Capacity and Total Phenol Content in Hop and Malt Commercial Samples

The antioxidant properties of commercial hops and malted barley were evaluated by electroanalytical methods and compared with spectrophotometric methods. Hop group showed better antioxidant capacity and total phenols content (TPC) values than malt group. The highest electrochemical Index (EI) was observed for HCO hop sample. Electrochemical Index (EI) showed a good Pearson correlation with ABTS (2,2′‐azino‐bis (3‐ethylbenzothiazoline‐6‐sulphonic acid)), DPPH (2,2‐diphenyl‐1‐picrylhydrazyl), and TPC, being of 0.85, 0.77 and 0.86 respectively for hop samples. While for the malt samples were 0.96, 0.69 and 0.49, respectively. The samples were compared to each other within each group and statistical differences can be observed, p<0.05. Therefore, the electroanalytical tools showed to be practical and easy to evaluate the antioxidant properties of hop and malt samples. The results may contribute to the selection of raw material for brewing and therefore with the quality of one of beverage one most consumed around the world.     

Assessment of the Antioxidant Capacity of Standard Compounds and Fruit Juices by a Newly Developed Electrochemical Method: Comparative Study with Results from Other Analytical Methods

The antioxidant capacity (AOC) of some standard compounds and commercial beverages was determined using an electroanalytical method which measures the OH radical scavenging ability (OH‐RSC). It is based on the degradation of a thin polyphenol film electrodeposited on glassy carbon following the attack by OH radicals produced by hydrogen peroxide photolysis. The degradation is prevented in presence of increasing concentrations of antioxidants. The same samples were also analyzed by DPPH, ABTS, and ORAC assays. The OH‐RSC method provided AOC values highly correlated with those resulting from the application of ORAC assay for both juices (R=0.90) and standard antioxidants (R=0.96).     

Chemical composition and in vitro antifungal and antioxidant activity of the essential oil and methanolic extract of Teucrium sauvagei Le Houerou

The chemical composition and the in vitro antifungal and antioxidant activity of the essential oil and the methanolic leaf extracts of Teucrium sauvagei Le Houerou, an endemic medicinal plant growing in Tunisia, have been studied. More than 35 constituents having an abundance >or=0.2% were identified in the oil. beta-Eudesmol, T-cadinol, alpha-thujene, gamma-cadinene, and sabinene were the prevalent constituents. Results of the antifungal activity tests indicated that the methanolic extract inhibited the in vitro growth of seven dermatophytes, whereas the essential oil showed average inhibition against only three dermatophytes. In vitro antioxidant properties of the essential oil and the methanolic extract were determined by DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azinobis(3-ethylbenzothiazoline-6-sulphonic acid)) assays and compared to those of the synthetic antioxidant Trolox. Due to their antifungal and antioxidant properties, the essential oil and the methanolic extract of T. sauvagei may be of use as natural preservative ingredients in food and/or pharmaceutical industries.     

Antioxidant Interactions between S-allyl-L-cysteine and Polyphenols Using Interaction Index and Isobolographic Analysis

This work aims to study the antioxidant interactions between S-allyl-L-cysteine (SAC) and six natural polyphenols (quercetin, caffeic acid, sinapic acid, catechin, ferulic acid, and 3,4-dihydroxybenzoic acid) through the measurement of free-radical-scavenging activity of 1,1-diphenyl- 2-picryl-hydrazyl (DPPH), the radical-cation-scavenging activity of 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), and reducing power. Among the six natural polyphenols, caffeic acid showed the strongest synergistic effect with SAC according to DPPH and reducing power assays. Further investigations based on the results of interaction index and isobologram analysis showed that the antioxidant activity (DPPH, ABTS, and reducing power) of the combination of caffeic acid with SAC presented an increase with the raising of their individual concentrations in their mixture and along with a dose–response manner. The best synergistic effect between caffeic acid and SAC based on DPPH, ABTS, and reducing power assays were observed at the ratio of 1:20, 1:35, and 1:70, respectively. The excellent synergic antioxidant activity of the combination of caffeic acid with SAC in our study suggests SAC has a more broad and effective application prospects in food field.     

Ocimum campechianum Mill. from Amazonian Ecuador: Chemical Composition and Biological Activities of Extracts and Their Main Constituents (Eugenol and Rosmarinic Acid)

The essential oil (EO), the methanolic (MeOH), and the 70% ethanolic (70% EtOH) extracts obtained from the aerial parts of Ocimum campechianum Mill. (Ecuador) were chemically characterized through gas-chromatography coupled to mass spectrometry detector (GC-MS), high-performance liquid chromatography coupled to diode array-mass spectrometry detectors (HPLC-DAD-MS) and studied for their in vitro biological activity. The radical scavenger activity, performed by spectrophotometric 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays, highlighted significant IC₅₀ values for the EO, extracts and their main constituents (eugenol and rosmarinic acid). EO (and eugenol) showed noteworthy activity against Pseudomonas syringae pv. syringae and a moderate effect against clinical Candida strains, with possible synergism in association to fluconazole against the latter microorganisms. The extracts and pure molecules exhibited weak cytotoxic activity against the HaCat cell line and no mutagenicity against Salmonella typhimurium TA98 and TA100 strains, giving indication of safety. Instead, EO showed a weak activity against adenocarcinomic human alveolar basal epithelial cells (A549). The above-mentioned evidence leads us to suggest a potential use of the crude drug, extracts, and EO in cosmetic formulation and food supplements as antioxidant agents. In addition, EO may also have a possible application in plant protection and anti-Candida formulations.     

Evaluation of pesticide residue in grape juices and the effect of natural antioxidants on their degradation rate

Various studies have been drawn toward the beneficial properties of fruit juices because they have several components, such as phenols, vitamins, and flavonoids, with antioxidant effects. However, fruit juices can also contain residues of pesticides used as standard pest control methods in crops. Many of these pesticides are degraded through oxidative mechanisms, and their persistence in juices can be enhanced by antioxidants. This study covers the degradation of four pesticides, aldicarb, demeton-S-methyl, fenamiphos, and methiocarb, to their respective sulfoxide and sulfone in grape juices, water (pH 3.5) and water (pH 3.5) with quercetin (one of the most important flavonoids of grape) added in an attempt to establish whether the presence of antioxidants can affect the degradation rate of pesticides. For this purpose, a multiresidue method based on solid-phase extraction (SPE) was developed for the simultaneous determination of these pesticides and their metabolites in commercial juices. The extraction procedure was carried out in C₁₈ columns. The subsequent elution of pesticides was performed with dichloromethane prior to the determination by liquid chromatography-tandem mass spectrometry (LC-MS/MS), using two precursor-product ion transitions. Average recoveries for all the pesticides studied were higher than 80%, with relative standard deviations lower than 15% in the concentration range 0.005–0.05 μg/mL, and the quantification limits achieved ranged from 0.1 to 4.6 μg/L. The results demonstrated that degradation was slower in fruit juices and aqueous solutions with quercetin than in water. Several commercial grape juices were also analyzed to establish the levels of these pesticides. Methiocarb, fenamiphos, and demeton-S-methyl were found at low levels in some samples.     

Assessment of antioxidant activities of three wild medicinal plants from Bahrain

This study reports the antioxidant properties of three wild medicinal plants from Bahrain, namely Aizoon canariense L., Asphodelus tenuifolius Cav., and Emex spinosus (L.) Campdera. Antioxidant and antiradical activities of dried materials of these plants were investigated using FRAP, DPPH and ABTS assays. Total phenolics, free phenolics and total flavonoids were also determined. E. spinosus was ranked by the assays as the plant possessing the highest antioxidant and antiradical activities with an average FRAP value of 1.84 mmol/g and IC₅₀ of 10.7 and 7.75 mg/ml for DPPH and ABTS assays, respectively. A. tenuifolius ranked second with a mean FRAP value of 0.69, IC₅₀ DPPH of 1.72 and ABTS of 0.36. A. canariense possessed the lowest activities with a mean FRAP value of 0.6, and averaged IC₅₀ of 103.8 and ABTS of 14.6. E. spinosus possessed the highest content of free phenolics (mg/100 g) (64.64) followed by A. tenuifolius (45.21) and A. canariense (32.23). E. spinosus also exhibited the highest total flavonoids with an average 82.71 mg/100 g followed by A. canariense (55.92) and A. tenuifolius (49.10). The studied medicinal plants possess considerable antioxidant activities and may contribute to the well-being of individuals who consume them.     

Common Trends and Differences in Antioxidant Activity Analysis of Phenolic Substances Using Single Electron Transfer Based Assays

Numerous assays were developed to measure the antioxidant activity, but each has limitations and the results obtained by different methods are not always comparable. Popular examples are the DPPH and ABTS assay. Our aim was to study similarities and differences of these two assay regarding the measured antioxidant potentials of 24 phenolic compounds using the same measurement and evaluation methods. This should allow conclusions to be drawn as to whether one of the assays is more suitable for measuring specific subgroups like phenolic acids, flavonols, flavanones, dihydrochalcones or flavanols. The assays showed common trends for the mean values of most of the subgroups. Some dihydrochalcones and flavanones did not react with the DPPH radical in contrast to the ABTS radical, leading to significant differences. Therefore, to determine the antioxidant potential of dihydrochalcone or flavanone-rich extracts, the ABTS assay should be preferred. We found that the results of the flavonoids in the DPPH assay were dependent on the Bors criteria, whereas the structure–activity relationship in the ABTS assay was not clear. For the phenolic acids, the results in the ABTS assay were only high for pyrogallol structures, while the DPPH assay was mainly determined by the number of OH groups.     

Free radical scavenging activities of naturally occurring and synthetic analogues of sea urchin naphthazarin pigments

Antioxidant activities of minor pigments of sea urchins (1-5) and synthetic naphthazarins (7-13) were evaluated and compared with echinochrome A (6) using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) scavenging assays. Structure-activity relationships showed that the antioxidant activities of the tested compounds depended on the number and positions of hydroxyl groups. Compounds bearing 3 or 2 hydroxyl groups on a naphthazarin core (5,8-dihydroxy-1,4-naphthoquinone) were the most active in both assays. Echinochrome A (6) (IC50 7.0 microM) and its monomethyl ethers 7 (IC50 15.0 microM) and 8 (IC50 15.0 microM) displayed stronger activities than Trolox (IC50 16.0 microM) in the DPPH and ABTS assays (TE = 3.41, 2.35, and 2.35 mM, respectively). Compounds with either one or without hydroxyl groups on a naphthazarin core displayed activities significantly lower than Trolox in both assays. These results suggest that hydroxylated naphthazarin pigments of sea urchins have a potential use as natural antioxidants.