2-2--azino-di-(3-ethylbenzothiazoline)-6-sulfonic-acid and 6-hydroxy-2-5-7-8-tetramethylchroman-2-carboxylic-acid

Topics: Antimalarials; Antioxidants; Benzothiazoles; Biphenyl Compounds; Chloroquine; Chromans; Parasitic Sensitivity Tests; Peroxides; Picrates; Plasmodium falciparum; Sulfonic Acids

This work aimed at investigating the behaviour of Trolox, vitamin E analogue, in presence of macromolecule-bound antioxidants in aqueous radical medium. Three main groups of macromolecule-bound antioxidants were assayed: dietary fiber (DF), protein and lipid-bound antioxidants, represented by whole wheat, soybean and olive oil products, respectively. Experimental studies were carried out in aqueous ABTS (2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)) radical medium. Trolox and macromolecule-bound antioxidants were added to radical separately and together in different concentrations. Antioxidant capacities were determined using QUENCHER procedure. pH of radical media was altered for DF and protein-bound antioxidant studies to examine its effect. Chemometric tools were used for experimental design and multivariate data analysis. Results revealed antagonistic interactions for Trolox with all macromolecule-bound antioxidants. The reason behind this antagonism was investigated through oxidation reactions of Trolox via mass spectrometry analysis. Consequently, a proof was obtained for inhibitory effect of bound-antioxidants on auto-regeneration reactions of Trolox.

Topics: Antioxidants; Benzothiazoles; Chromans; Glycine max; Olive Oil; Oxidation-Reduction; Plant Proteins; Sulfonic Acids; Triticum; Vitamin E

Apples (Malus domestica L.) are the most common source of phenolic compounds in northern European diet. Besides pectins, dietary fibers, vitamins, and oligosaccharides they contain phenolic compounds of different classes. Apple powders are convenient functional forms retaining significant amounts of phenolic antioxidants. In this study reducing and radical scavenging profiles of freeze-dried powders of "Aldas,ˮ "Auksis,ˮ "Connel Red,ˮ "Ligol,ˮ "Lodel,ˮ and "Rajkaˮ were determined and phenolic constituents were identified using ultra high-performance liquid chromatography coupled to quadrupole and time-of-flight mass spectrometers. A negative ionization mode was applied and seventeen compounds: phenolic acids (coumaroylquinic, chlorogenic), flavonoids (quercetin derivatives), and procyanidin derivatives (B1, B2, and C1) were identified in all tested apple samples. Total values of Trolox equivalents varied from 7.72 ± 0.32 up to 20.02 ± 0.52 and from 11.10 ± 0.57 up to 21.42 ± 0.75 μmol/g of dry weight of apple powder in FRAP (ferric reducing antioxidant power) and ABTS (2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) postcolumn assays, respectively. The greatest Trolox equivalent values were determined for apples of "Aldasˮ cultivar. Chlorogenic acid and procyanidin C1 were the most significant contributors to total reducing and radical scavenging activity in all apple cultivars tested, therefore they could be considered as markers of antioxidant activity.

Topics: Antioxidants; Benzothiazoles; Biflavonoids; Catechin; Chlorogenic Acid; Chromans; Flavonoids; Fruit; Humans; Malus; Oxidation-Reduction; Phenols; Plant Extracts; Powders; Proanthocyanidins; Quercetin; Sulfonic Acids

Eight phenolic compounds including: p-coumaric acid, vanillic acid, caffeic acid, chlorogenic acid, trolox, quercetin, curcumin, and resveratrol were treated with riboflavin (RF) photosensitization and in vitro antioxidant capacities of the mixtures were determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2' azino bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) assays. Mixtures containing p-coumaric acid and vanillic acid under RF photosensitization showed increases in ferric ion reducing ability and radical scavenging activity of DPPH, whereas mixtures of other compounds had decreases in both radical scavenging ability and ferric reducing antioxidant power. Hydroxycoumaric acid and conjugated hydroxycoumaric and coumaric acids were tentatively identified from RF photosensitized p-coumaric acid, whereas dimmers of vanillic acid were tentatively identified from RF photosensitized vanillic acid. RF photosensitization may be a useful method to enhance antioxidant properties like ferric ion reducing abilities of some selected phenolic compounds.

Topics: Antioxidants; Benzothiazoles; Biphenyl Compounds; Caffeic Acids; Chlorogenic Acid; Chromans; Coumaric Acids; Curcumin; Light; Molecular Structure; Oxidation-Reduction; Phenols; Picrates; Plant Extracts; Propionates; Quercetin; Resveratrol; Riboflavin; Stilbenes; Sulfonic Acids; Vanillic Acid

Topics: Antioxidants; Benzothiazoles; Biphenyl Compounds; Butylated Hydroxyanisole; Chromans; Coprinus; Culture Media; Inhibitory Concentration 50; Molecular Structure; Picrates; Sesquiterpenes; Sulfonic Acids

Topics: Antineoplastic Agents; Antioxidants; Benzothiazoles; Biological Assay; Biphenyl Compounds; Chromans; Drug Screening Assays, Antitumor; Free Radical Scavengers; Humans; MCF-7 Cells; Picrates; Sulfonic Acids; Taurine; Tumor Cells, Cultured

Using a TLC autographic assay for radical-scavenging activity with the ABTS radical, the presence of lipophilic antioxidants in the larvae of the wild bruchid seed beetle Bruchidius dorsalis was detected. Assay-guided fractionation of the CHCl3-soluble fraction of the larvae resulted in the isolation of new glycerolipids, designated dorsamin-A763, -A737, -A765, -A739, and -A767, comprising 1,2-diacyl-sn-glycero-3-dehydrophenylalanine ester structural units. The ABTS radical scavenging activity of the dorsamin-A's was comparable with or stronger than that of Trolox.

Topics: Animals; Antioxidants; Benzothiazoles; Chromans; Coleoptera; Feeding Behavior; Glycolipids; Larva; Molecular Structure; Phenylalanine; Reactive Oxygen Species; Sulfonic Acids

Reaction kinetics in the Trolox equivalent antioxidant capacity (TEAC) assay between ABTS(+•) [2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) cation radical] and compounds with different structure, molecular weight, number of OH groups, and redox potential were investigated by recording loss of ABTS(+•) absorbance (734 nm) continuously over time. Curves showed six distinguishable kinetic patterns, including both immediate and extended reaction components. Radical quenching rates in the immediate component most relevant to reactions in foods and tissues depended on phenol structure and steric accessibility to the hindered radical, while reaction stoichiometry correlated with the number of phenol groups (>0.81) but not redox potential. Current assay procedures measure antioxidant capacity under conditions not relevant to actual applications and do not determine radical quenching rates. Results raise serious questions regarding the ability of reactions with the hindered ABTS(+•) to rank actual radical quenching by compounds with different structures and invalidate reporting antioxidant activity as Trolox equivalents.

Topics: Antioxidants; Benzothiazoles; Chromans; Kinetics; Molecular Structure; Oxidation-Reduction; Sulfonic Acids

Alpinia oxyphylla Miquel (Zingiberaceae) is a traditional Chinese herbal medicine widely used for the treatment of intestinal disorders, urosis and diuresis. However, information about antioxidant and cytotoxic properties of its fruits remains to be elucidated.. The ethanol crude extract (CE) and its fractions [petroleum ether fraction (PF), ethyl acetate fraction (EF), n-butanol fraction (BF) and water fraction (WF) extracted by petroleum ether, ethyl acetate, n-butanol and water, respectively] of A. oxyphylla fruits were investigated for their antioxidant activity and cytotoxicity.. The total phenolic content (TPC) and antioxidant activity of the extracts were determined by Folin-Ciocalteu reagent, 1,1-diphenyl-2-picrylhydrazyl (DPPH(•)), Trolox equivalent antioxidant capacity and reducing power assay. Cytotoxicity of the extracts (0-200 μg/mL) was tested on six human cancer cell lines (breast cancer cell line, cervix carcinoma cell line, lung adenocarcinoma cell line, liver carcinoma cell line, gastric cancer cell line and colon cancer cell line) using the sulforhodamine B assay.. The TPC of extracts varied from 8.2 to 20.3 mg gallic acid equivalents/g dry weight. DPPH radical scavenging effect of extracts decreased in the order of EF > BF > CE > PF > WF, with IC50 values ranging from 74.7 to 680.8 μg/mL. 2,2-azo-bis(3-Ethylbenzothiazoline-6-sulfoic acid) diammonium salt scavenging activity ranged from 0.118 to 0.236 mmol Trolox equivalence/mg extract. The extracts exhibited concentration-dependent reducing power, and EF showed the highest reducing ability. A satisfactory correlation (R(2) > 0.826) between TPC and antioxidant activity was observed. In addition, EF, PF and CE exhibited potent anticancer effects on six cancer cell lines with IC50 values ranging from 40.1 to 166.3 μg/mL.. The ethanol extract of A. oxyphylla fruit, especially the EF, was found to possess potent antioxidant and anticancer activities, and thus a great potential for the application in food and drug products.

Topics: Alpinia; Antineoplastic Agents, Phytogenic; Antioxidants; Benzothiazoles; Biphenyl Compounds; Cell Line, Tumor; Cell Proliferation; Cell Survival; Chromans; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Ethanol; Fruit; Humans; Inhibitory Concentration 50; Oxidation-Reduction; Phenols; Phytotherapy; Picrates; Plant Extracts; Plants, Medicinal; Solvents; Sulfonic Acids

We studied antioxidant activity of new derivative of pyrrolo[1,2-α]benzimidazole RU-792 and compared its effects on free radical processes with those of the reference antioxidant Trolox in four model free-radical systems. RU-792 had high antioxidant activity determined by its intrinsic antiradical properties. RU-792 was superior to Trolox by antioxidant activity in the models of Fe(2+)-induced chemiluminescence of lipids with 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid), but less effective in the model of luminol-dependent chemiluminescence.

Topics: Benzimidazoles; Benzothiazoles; Biphenyl Compounds; Chromans; Free Radical Scavengers; Free Radicals; Kinetics; Lipid Peroxidation; Oxidation-Reduction; Picrates; Pyrroles; Sulfonic Acids

Accurate monitoring of the antioxidant status or of oxidative stress in patients is still a big challenge in clinical laboratories. This study investigates the possibility of applying a newly developed total antioxidant capacity assay method based on laccase or peroxidase oxidized syringaldazine [Tetramethoxy azobismethylene quinone (TMAMQ)] which is referred to here as SyrinOX, as a diagnostic tool for monitoring both oxidative stress and antioxidant status in patients. Attempts to adapt the Randox total antioxidant procedure [simultaneous incubation of the radical generating system (metmyoglobin and H(2) O(2) ) and antioxidant sample] for SyrinOX were abandoned after it was discovered that the H(2) O(2) reacted with enzymatically generated TMAMQ and ABTS radicals at a rate of 6.4 × 10(-2) /μM/s and 5.7 × 10(-3) /μM/s respectively. Thus this study for the first time demonstrates the negative effects of H(2) O(2) in the Randox system. This leads to erroneous results because the total antioxidant values obtained are the sum of radicals reduced by antioxidants plus those reacting with the radical generating system. Therefore they should be avoided not only for this particular method but also when using other similar methods. Consequently, SyrinOX is best applied using a three-step approach involving, production of TMAMQ, recovery and purification (free from enzyme and other impurities) and then using TMAMQ for measuring the total antioxidant capacity of samples. Using this approach, the reaction conditions for application of SyrinOX when measuring the total antioxidant capacity of plasma sample were determined to be 50% (v/v) ethanol/50 mM sodium succinate buffer pH 5.5, between 20 and 25 °C for at least 1 h.

Topics: Antioxidants; Benzothiazoles; Biological Assay; Biomarkers; Buffers; Calibration; Chromans; Ethanol; Humans; Hydrazones; Hydrogen Peroxide; Hydrogen-Ion Concentration; Indicators and Reagents; Kinetics; Metmyoglobin; Oxidation-Reduction; Oxidative Stress; Predictive Value of Tests; Reference Standards; Reproducibility of Results; Solvents; Spectrum Analysis; Succinic Acid; Sulfonic Acids; Temperature; Thiazoles

Substantial evidence exists to support the hypothesis that high fruit and vegetable consumption, rich in antioxidants, can reduce the incidence of several disease states. The aim of this study was to compare the results obtained by six spectrophotometric biochemical methods including the ferric reducing antioxidant power (FRAP), 2, 2-diphenyl-1-picryhydrazyl (DPPH•), 2,2-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS•⁺), copper (II) reducing capacity (CUPRAC) and Cerium (IV) reducing antioxidant capacity (CERAC) assays as well as Folin-Ciocalteu method (FC) for the measurement of total antioxidant capacity (TAC) and total polyphenols (TP) in different commercially available vegetable juices. There was a significant positive correlation between the results obtained for FRAP, ABTS•⁺, CUPRAC, CERAC and FC (0.68 ≤ r ≤ 0.96, P < 0.01). DPPH• was only correlated with CERAC (r = 0.66, P < 0.01). Beetroot juice had the highest TAC and TP regardless of the method of analysis.

Topics: Antioxidants; Benzothiazoles; Beverages; Biphenyl Compounds; Chromans; Fluorescence Recovery After Photobleaching; Picrates; Polyphenols; Spectrophotometry; Sulfonic Acids; United Kingdom; Vegetables

Vanillin, a compound widely used in foods, beverages, cosmetics and drugs, has been reported to exhibit multifunctional effects such as antimutagenic, antiangiogenetic, anti-colitis, anti-sickling, and antianalgesic effects. However, results of studies on the antioxidant activity of vanillin are not consistent.. We systematically evaluated the antioxidant activity of vanillin using multiple assay systems. DPPH radical-, galvinoxyl radical-, and ABTS(+)-scavenging assays, ORAC assay and an oxidative hemolysis inhibition assay (OxHLIA) were used for determining the antioxidant activity.. Vanillin showed stronger activity than did ascorbic acid and Trolox in the ABTS(+)-scavenging assay but showed no activity in the DPPH radical- and galvinoxyl radical-scavenging assays. Vanillin showed much stronger antioxidant activity than did ascorbic acid and Trolox in the ORAC assay and OxHLIA. In the ABTS(+)-scavenging assay, ORAC assay and OxHLIA, vanillin reacted with radicals via a self-dimerization mechanism. The dimerization contributed to the high reaction stoichiometry against ABTS(+) and AAPH-derived radicals to result in the strong effect of vanillin. Oral administration of vanillin to mice increased the vanillin concentration and the antioxidant activity in plasma. These data suggested that antioxidant activity of vanillin might be more beneficial than has been thought for daily health care.. Based on the results of the present study, we propose the addition of antioxidant capacity to the multifunctionality of vanillin.

Topics: Animals; Antioxidants; Ascorbic Acid; Benzaldehydes; Benzhydryl Compounds; Benzothiazoles; Biphenyl Compounds; Chromans; Chromatography, High Pressure Liquid; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical; Erythrocytes; Free Radical Scavengers; Hemolysis; Mice; Molecular Structure; Oxidation-Reduction; Picrates; Sheep; Sulfonic Acids

This study reports the antioxidant and antibacterial activities of four fresh mango seed extracts from Thai varieties. Total phenol contents determined by the Folin-ciocalteu method revealed the highest values to be in MKE, Chok-a-nan variety (399.8 mgGAE/g extract) and MSE of Nam-dok-mai variety (377.2 mgGAE/g extract). Both extracts showed potent ABTS˙+ radical and DPPH˙ radical scavenging activities with the lower half inhibition concentration (IC50) values than those of the reference compounds; vitamin C, trolox and BHA, respectively. Their antioxidant property of MSE and MKE is strongly correlated with the total phenol contents (r=0.98 and 0.98, respectively). When combined the MSE and MKE of the Fah-lun variety showed the strongest antioxidant activity. All mango seed extracts showed interesting antibacterial activity against both gram positive and gram negative bacteria as determined by disc diffusion method. The most sensitive pathogenic strain inhibited by all extracts (especially Kaew variety) was Pseudomonas aeruginosa ATCC 27853. This work suggests potential applications for practical uses of mango seed extracts from Thai varieties, as sources of antioxidant and antibacterial agents.

Topics: Anti-Bacterial Agents; Antioxidants; Ascorbic Acid; Bacteria; Benzothiazoles; Biphenyl Compounds; Chromans; Diffusion; Free Radical Scavengers; Indicators and Reagents; Mangifera; Microbial Sensitivity Tests; Phenols; Picrates; Plant Extracts; Seeds; Sulfonic Acids; Thailand

Cellular damage induced by free-radicals like Reactive Oxygen and Nitrogen Species (ROS and RNS) has been implicated in several disorders and diseases, including ageing. Hence naturally occurring anti-oxidant rich-herbs play a vital role in combating these conditions. The present study was carried out to investigate the in vitro free-radical quenching capacity of a known Ayurvedic poly-herbal formulation called Vayasthapana Rasayana.. Methanol extracts of Vayasthapana Rasayana formulation (VRF) were studied for in vitro total antioxidant activity along with phenolic content and reducing power. In vitro assays like DPPH, FRAP, ABTS scavenging to evaluate radical quenching potential were performed.. The formulation has shown 94% at 0.1 mg/ml DPPH free-radical scavenging activity as against 84% at 0.1 mg/ml for standard ascorbic acid (IC₅₀ value 5.51 μg/ml for VRF and 39 μg/ml for standard). It has a significant higher ferric reducing potential also (OD 0.87 at 700 nm & 0.21 at 0.1 mg/ml for VRF and standard, respectively). The total phenolic content (gallic acid equivalent) of the VRF is 8.3 mg per g of dry mass. Total antioxidant capacity of the formulation, estimated by FRAP was 1150 ± 5 μM Fe(II)/g dry mass. ABTS radical scavenging activity of VRF was 69.55 ± 0.21% at 100 μg/ml concentration with a IC50 value of 69.87 μg/ml as against 9% and 95% by ascorbic acid and Trolox (at 70.452 μg/ml and 0.250 μg/ml concentrations, respectively).. In Indian traditional Ayurvedic system, use of VRF is in regular practice for mainly combating age-related disorders and diseases as many of the components of the Rasayana are known for their free-radical scavenging activity. This study has validated the potential use of VRF as an anti-oxidant to fight age-related problems.

Topics: Aging; Antioxidants; Ascorbic Acid; Benzothiazoles; Biphenyl Compounds; Chromans; Ferric Compounds; Humans; Magnoliopsida; Medicine, Ayurvedic; Oxidative Stress; Phenols; Picrates; Plant Extracts; Reference Standards; Sulfonic Acids; Thiazoles

Ramalin (γ-glutamyl-N'-(2-hydroxyphenyl)hydrazide), a novel compound, was isolated from the methanol-water extract of the Antarctic lichen Ramalina terebrata by several chromatographic methods. The molecular structure of ramalin was determined by spectroscopic analysis. The experimental data showed that ramalin was five times more potent than commercial butylated hydroxyanisole (BHA) in scavenging 1-diphenyl-2-picryl-hydazil (DPPH) free radicals, 27 times more potent in scavenging 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid free radicals (ABTS(+)) than the vitamin E analogue, trolox, and 2.5 times more potent than BHT in reducing Fe(3+) to Fe(2+) ions. Similarly, ramalin was 1.2 times more potent than ascorbic acid in scavenging superoxide radicals and 1.25 times more potent than commercial kojic acid in inhibiting tyrosinase enzyme activity, which ultimately leads to whitening of skin cells. Ramalin showed no or very little cytotoxicity in human keratinocyte and fibroblast cells at its antioxidant concentration. Furthermore, ramalin was assessed to determine its antioxidant activity in vivo. One microgram per milliliter ramalin significantly reduced the released nitric oxide (NO) and 0.125 μg/ml ramalin reduced the produced hydrogen peroxide (H(2)O(2)) in LPS (lipopolysaccharide)-stimulated murine macrophage Raw264.7 cells. Considering all the data together, ramalin can be a strong therapeutic candidate for controlling oxidative stress in cells.

Topics: Animals; Antarctic Regions; Antioxidants; Ascorbic Acid; Benzothiazoles; Biphenyl Compounds; Butylated Hydroxyanisole; Cell Line; Cell Survival; Chromans; Fibroblasts; Free Radicals; Fungal Proteins; Glutamates; Humans; Hydrogen Peroxide; Keratinocytes; Lichens; Mice; Molecular Structure; Monophenol Monooxygenase; Nitric Oxide; Picrates; Pyrones; Sulfonic Acids

We investigated in vitro antioxidant activities of 49 endophytic fungi isolated from the liverwort Scapania verrucosa. Based on morphological and molecular identification, the endophytic fungi isolated were classified into seven genera (Hypocrea, Penicillium, Tolypocladium, Chaetomium, Xylaria, Nemania, and Creosphaeria), all belonging to one family (Xylariaceae). By screening with the 2,2'-azino-di(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) decolorization assay, the ethyl acetate extracts of five endophytic fungi (T7, T21, T24, T32, and T38 strains), which exhibited remarkable Trolox equivalent (TE) antioxidant capacity (ranging from 997.06 to 1248.10 μmol TE/g extract), were selected and their antioxidant capacity was further evaluated by assays for 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, hydroxyl radical scavenging, reducing power, and ferrous ion chelating. The ethyl acetate extracts of two endophytic fungi (T24 and T38) were found to have comparable scavenging abilities on both DPPH-free radicals (93.9 and 88.7%, respectively, at 50 μg/mL) and hydroxyl radicals (97.1 and 89.4%, respectively, at 2 mg/mL) when compared with those of the positive controls (ascorbic acid and butylated hydroxytoluene, respectively). Although their reducing powers were similar to that of butylated hydroxytoluene, as indicated by absorbance (0.35 and 0.30 at 50 μg/mL, respectively), only the T38 strain's ethyl acetate extract showed ferrous ion chelating ability (92.9% at 1 mg/mL) comparable to that of the EDTA-2Na control. These endophytic fungi in S. verrucosa are a potential novel source of natural antioxidants.

Topics: Acetates; Antioxidants; Benzothiazoles; Biphenyl Compounds; China; Chromans; Complex Mixtures; Endophytes; Free Radical Scavengers; Free Radicals; Fungi; Genes, Fungal; Genes, rRNA; Hepatophyta; Hydroxyl Radical; Iron Chelating Agents; Oxidation-Reduction; Picrates; Solvents; Sulfonic Acids

Antioxidant and antimicrobial activities, as well as total phenol (TP, Folin-Ciocalteu method) and phenolic acid (UPLC-MS/MS) contents of leaf and flower infusions of Teucrium arduini L. from six different mountainous localities in Croatia (Ucka, Vosac, Sveti Jure, Snjeznica, Vaganac, Susanj) were analysed in this study. Antioxidant capacity was evaluated using the ferric reducing/antioxidant power (FRAP) assay, as well as 2,2-diphenyl-1-picrylhydrazyl (DPPH), and 2,2'-azinobis (3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radical scavenging assays. The antioxidant potency composite index (ACI), giving equal weight to all three methods used to quantify antioxidant capacity, was the highest for the sample from Vosac (96.7) among flower infusions, while maximum ACI (100) was determined for the infusion from Ucka among leaf infusions. Strong positive correlation was found between the total phenols and ACI for leaf (r=0.953) and flower (r=0.977) infusions. Our results point to significantly (p<0.05) different TP content between leaf and flower infusions, as well as across localities. Leaf infusions of T. arduini from Susanj exhibited marked antibacterial activity against Staphylococcus aureus, while none of the tested infusions exhibited antimicrobial activity against gram-negative bacterial species, or the tested fungal species.

Topics: Anti-Infective Agents; Antioxidants; Bacteria; Benzothiazoles; Biphenyl Compounds; Chromans; Croatia; Ferric Compounds; Flowers; Free Radical Scavengers; Fungi; Glycosides; Indicators and Reagents; Microbial Sensitivity Tests; Phenols; Picrates; Plant Extracts; Plant Leaves; Reducing Agents; Sulfonic Acids; Teucrium

In this study, 24 Wistar rats were allocated to 4 groups of 6 animals each. Groups 1 and 2 were fed a basal diet, while groups 3 and 4 were fed the basal diet supplemented further with ground rosemary at 1% level. Following 6-weeks feeding, groups 2 and 4 were injected 1 ml CCl(4)/kg bw and after six hours all animals were sacrificed. Results showed that feeding rosemary before CCl(4) treatment resulted in decline (P<0.05) of the increased aspartate transaminase, alanine transaminase and alkaline phosphatase activities and increase (P<0.05) of the reduced cholesterol and triacylglycerols in serum. It also decreased (P<0.05) lipid peroxidation and increased (P<0.05) the reduced hydroxyl anion radical and hydrogen peroxide scavenging activities in serum, liver, kidney and heart tissues. In addition, it increased (P<0.05) the reduced ABTS radical cation and the superoxide anion scavenging activities in all tissues except in heart and in kidney and heart tissues, respectively. These results suggest that dietary rosemary has the potential to become a promising functional food component.

Topics: Animals; Antioxidants; Benzothiazoles; Carbon Tetrachloride Poisoning; Chromans; Diet; Electron Transport; Female; Free Radicals; Kidney; Ledum; Lipid Peroxidation; Liver; Malondialdehyde; Myocardium; NADPH-Ferrihemoprotein Reductase; Phenols; Plant Leaves; Rats; Rats, Wistar; Reactive Oxygen Species; Sulfonic Acids

The influence of genotype and climatic factors, e.g. mean temperature and mean global radiation level, on the antioxidant activity of kale was investigated. Therefore, eight kale cultivars, hybrid and traditional, old cultivars, were grown in a field experiment and harvested at four different times. In addition to the investigation of the total phenolic content, the overall antioxidant activity was determined by TEAC assay and electron spin resonance spectrometry. A special aim was to characterize the contribution of single flavonoids to the overall antioxidant activity using an HPLC-online TEAC approach. The antioxidant activity and the total phenolic content were influenced by the genotype and the eco-physiological factors. The HPLC-online TEAC results showed that not all flavonol glycosides contribute to the overall antioxidant activity in the same manner. Taking the results of the structural analysis obtained by HPLC-ESI-MS(n) into account, distinct structure-antioxidant relationships have been observed.

Topics: Antioxidants; Benzothiazoles; Brassica; Chromans; Chromatography, High Pressure Liquid; Climate; Colorimetry; Electron Spin Resonance Spectroscopy; Flavonoids; Free Radical Scavengers; Genotype; Phenols; Spectrometry, Mass, Electrospray Ionization; Sulfonic Acids

Antioxidants are health beneficial compounds that can protect cells and macromolecules (e.g., fats, lipids, proteins, and DNA) from the damage of reactive oxygen species (ROS). Solvent effect is a crucial parameter on the chemical behaviour of antioxidant compounds but there has been limited information regarding its role on antioxidant capacity and its assays. Therefore, the present study was undertaken to investigate the total antioxidant capacity (TAC) of some certain lipophilic and hydrophilic antioxidants, measured in different solvent media such as ethanol (EtOH) (100%), methanol (MeOH) (100%), methanol/water (4:1, v/v), methanol/water (1:1, v/v), dichloromethane (DCM)/EtOH (9:1, v/v). The cupric reducing antioxidant capacity (CUPRAC) values of selected antioxidants were experimentally reported in this work as trolox equivalent antioxidant capacity (TEAC), and compared to those found by reference TAC assays, i.e., 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)/persulphate (ABTS/persulphate) and ferric reducing antioxidant power (FRAP) methods. The TAC values of synthetic mixtures of antioxidants were experimentally measured as trolox equivalents and compared to those theoretically found by making use of the principle of additivity of absorbances assuming no chemical interaction between the mixture constituents. Possible synergistic (e.g., BHT and BHA in DCM/EtOH) or antagonistic behaviours of these synthetic mixtures were investigated in relation to solvent selection.

Topics: Anions; Antioxidants; Benzothiazoles; Chemistry Techniques, Analytical; Chromans; Copper; Hydrogen Bonding; Methanol; Methylene Chloride; Models, Chemical; Solvents; Sulfonic Acids; Tea; Water

The aim of the study was to analyse the relation between total antioxidant capacity and immunosuppressive therapies, renal function and hematocrit in kidney transplant patients. The study included 46 adult patients during the maintenance period (>1 year) following renal transplantation, treated with different combinations of immunosuppressive agents--most commonly with cyclosporine (n = 23) or tacrolimus (n = 15). The total antioxidant capacity (TAOC) of plasma was measured using Trolox-equivalent antioxidant capacity (TEAC) assay. Patients treated with cyclosporine had significantly greater TAOC compared with those treated with tacrolimus (1.16 +/- 0.46 mmol/L vs. 0.80 +/- 0.37 mmol/L, p = 0.018, respectively). There was a significantly negative correlation between TAOC and plasma creatinine (rs = -0.551, p = 0.033) and a positive correlation between TAOC and creatinine clearance or hematocrit in patients treated with tacrolimus but not with cyclosporine (r = 0.525, p = 0.045 or rs = 0.629, p = 0.012, respectively). Immunosuppressive therapy with cyclosporine was associated with higher TAOC. Anemia can be an independent risk factor for an increase of oxidative stress. Although subject numbers werelimited, TAOC was positively associated with renal function in patients treated with tacrolimus.

Topics: Adult; Aged; Antioxidants; Benzothiazoles; Calibration; Chromans; Creatinine; Cyclosporine; Diabetes Mellitus; Female; Hematocrit; Humans; Immunosuppressive Agents; Indicators and Reagents; Kidney Function Tests; Kidney Transplantation; Male; Middle Aged; Sulfonic Acids; Young Adult

The antioxidant activity of a novel synthetic selenadiazole derivative SPO against DPPH and ABTS free radicals was evaluated using spectrometric methods. The results show that the detection wavelength and stable time for DPPH system were 515 nm and 30 min respectively, while those for ABTS system were 734 nm and 6 min, respectively. SPO could effectively and rapidly inhibited the formation of ABTS and DPPH free radicals in a dose- and time-dependent manner, indicating the potent antioxidant activity of SPO under both hydrophilic and hydrophobic conditions. In the optimized systems, the IC50 values of SPO were 85.2 micromol x L(-1) (DPPH assay) and 36.5 micromol x L(-1) (ABTS assay), respectively, which were comparable with the standard antioxidant Trolox, and significantly better than the positive controls BHA and BHT. Taken together, our results suggest the potential applications of selenadiazole derivatives as antioxidative agents.

Topics: Antioxidants; Benzothiazoles; Biphenyl Compounds; Chromans; Free Radicals; Organoselenium Compounds; Oxidation-Reduction; Picrates; Spectrum Analysis; Sulfonic Acids

Comparison of the effectiveness of antioxidant activity of three thiol compounds, D-penicillamine, reduced L-glutathione, and 1,4-dithioerythritol, expressed as a radical-scavenging capacity based on the two independent methods, namely a decolorization 2,2'-azinobis[3-ethylbenzothiazoline-6-sulfonic acid] assay and a rotational viscometry, is reported. Particular concern was focused on the testing of potential free-radical scavenging effects of thiols against hyaluronan degradation, induced by hydroxyl radicals. A promising, solvent-independent, antioxidative function of 1,4-dithioerythritol, comparable to that of a standard compound, Trolox(®), was confirmed by the 2,2'-azinobis[3-ethylbenzothiazoline-6-sulfonic acid] assay. The new potential antioxidant 1,4-dithioerythritol exhibited very good solubility in a variety of solvents (e.g., H(2)O, EtOH, and DMSO) and could be widely accepted and used as an effective antioxidant standard instead of a routinely used Trolox(®) on 2,2'-azinobis[3-ethylbenzothiazoline-6-sulfonic acid] assay.

Topics: Antioxidants; Benzothiazoles; Chromans; Dithioerythritol; Drug Evaluation, Preclinical; Free Radical Scavengers; Glutathione; Hyaluronic Acid; Hydroxyl Radical; Oxidation-Reduction; Penicillamine; Solubility; Solvents; Spectrophotometry; Structure-Activity Relationship; Sulfhydryl Compounds; Sulfonic Acids; Thiazoles; Viscosity

L-adrenaline belongs to a group of the compounds known as catecholamines, which play an important role in the regulation of physiological process in living organisms. The antioxidant activity and antioxidant mechanism of L-adrenaline was clarified using various in vitro antioxidant assays including 1,1-diphenyl-2-picryl-hydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), N,N-dimethyl-p-phenylenediamine (DMPD(+)), and superoxide anion radicals (O(2)(-)) scavenging activity, hydrogen peroxide (H(2)O(2)), total antioxidant activity, ferric ions (Fe(3+)) and cupric ions (Cu(2+)) reducing ability, ferrous ions (Fe(2+)) chelating activity. L-adrenaline inhibited 74.2% lipid peroxidation of a linoleic acid emulsion at 30 microg/mL concentration. On the other hand, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), alpha-tocopherol and trolox displayed 83.3, 82.1, 68.1 and 81.3% inhibition on the peroxidation of linoleic acid emulsion at the same concentration, respectively. BHA, BHT, alpha-tocopherol and trolox were used as reference antioxidants and radical scavenger compounds. Moreover, this study will bring an innovation for further studies related to antioxidant properties of L-adrenaline. According to present study, L-adrenaline had effective in vitro antioxidant and radical scavenging activity.

Topics: alpha-Tocopherol; Antioxidants; Benzothiazoles; Biphenyl Compounds; Butylated Hydroxyanisole; Butylated Hydroxytoluene; Chromans; Copper; Dose-Response Relationship, Drug; Epinephrine; Ferrous Compounds; Free Radical Scavengers; Hydrogen Peroxide; Linoleic Acid; Lipid Peroxidation; Oxidation-Reduction; Picrates; Piperidones; Structure-Activity Relationship; Sulfonic Acids; Superoxides

We previously reported that treatment of spontaneously hypertensive rats (SHR) with liver growth factor (LGF), an albumin-bilirubin complex with a covalent bond, reduces blood pressure, improves nitric oxide (NO)-dependent vasodilatation, and exerts vascular antifibrotic actions. Because bilirubin, albumin, and albumin-bound bilirubins have antioxidant properties, we hypothesize that LGF might exert its cardiovascular actions through an antioxidant mechanism. We have tested in vitro the capacity of LGF to scavenge ABTS cation and peroxyl and hydroxyl radicals and to protect vascular NO from degradation by superoxide anion. We have also compared the antioxidant capacity of LGF with that of its molecular components albumin and bilirubin and the reference antioxidant trolox. LGF exhibited antioxidant capacity against all free radicals tested at lower concentrations than albumin, bilirubin, and trolox. LGF, bilirubin, and albumin were also able to protect endothelial NO from superoxide anion degradation in a fashion similar to that of superoxide dismutase or tiron, but at much lower concentrations. These data, together with our previous results in SHR, suggest that LGF might exert its cardiovascular regenerative actions, at least in part, through an antioxidant mechanism and that LGF could be a relevant circulating antioxidant in situations of oxidative stress.

Topics: Animals; Antioxidants; Benzothiazoles; Bilirubin; Blood Pressure Determination; Carotid Arteries; Chromans; Endothelial Cells; Fibrosis; Hydroxyl Radical; Hypertension; Male; Nitric Oxide; Oxidative Stress; Peroxides; Protein Binding; Rats; Rats, Sprague-Dawley; Serum Albumin; Serum Albumin, Bovine; Serum Albumin, Human; Sulfonic Acids

Radical scavenging activities of bovine milk components were quantified following size exclusion chromatography (SEC) with postcolumn characterization of fractions using the scavenging of 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) radicals (ABTS*(+)) in the Trolox equivalent antioxidant capacity (TEAC) assay and peroxyl radicals formed from cleavage of 2,2'-azobis(2-amidinopropane) (AAPH) in the oxygen radical absorbance capacity (ORAC) fluorometric assay. Caseins were quantitatively the major radical scavenger species in both assays, whereas beta-lactoglobulin (beta-lg) and alpha-lactalbumin (alpha-la) were much less active and only in the peroxyl radical assay. The radical scavenging activity of the caseins could be quantitatively accounted for by their constituent amino acids, as there were no effects of denaturing agents or complete digestion with proteases. In contrast, the activities of the whey proteins were dependent on denaturation or partial hydrolysis and dominated by the free thiol in beta-lg. A component in milk serum with a molecular mass of approximately 100 kDa contributed significantly to both ABTS*(+) and peroxyl radical scavenging but was absent in whey. This radical scavenger was identified as beta-casein. The only significant low molecular weight radical scavenger species were identified as ascorbate and urate in both assays.

Topics: Amino Acids; Animals; Antioxidants; Benzothiazoles; Caseins; Cattle; Chromans; Chromatography, Gel; Free Radical Scavengers; Lactalbumin; Lactoglobulins; Milk; Milk Proteins; Peroxides; Protein Folding; Sulfonic Acids; Whey Proteins

The influence of home cooking methods (boiling, microwaving, pressure-cooking, griddling, frying, and baking) on the antioxidant activity of vegetables has been evaluated in 20 vegetables, using different antioxidant activity assays (lipoperoxyl and hydroxyl radicals scavenging and TEAC). Artichoke was the only vegetable that kept its very high scavenging-lipoperoxyl radical capacity in all the cooking methods. The highest losses of LOO. scavenging capacity were observed in cauliflower after boiling and microwaving, pea after boiling, and zucchini after boiling and frying. Beetroot, green bean, and garlic kept their antioxidant activity after most cooking treatments. Swiss chard and pepper lost OH. scavenging capacity in all the processes. Celery increased its antioxidant capacity in all the cooking methods, except boiling when it lost 14%. Analysis of the ABTS radical scavenging capacity of the different vegetables showed that the highest losses occurred in garlic with all the methods, except microwaving. Among the vegetables that increased their TEAC values were green bean, celery, and carrot after all cooking methods (except green bean after boiling). These 3 types of vegetables showed a low ABTS radical scavenging capacity. According to the method of analysis chosen, griddling, microwave cooking, and baking alternately produce the lowest losses, while pressure-cooking and boiling lead to the greatest losses; frying occupies an intermediate position. In short, water is not the cook's best friend when it comes to preparing vegetables.

Topics: Antioxidants; Apium; Benzothiazoles; Chromans; Cooking; Daucus carota; Fabaceae; Free Radical Scavengers; Hot Temperature; Hydroxyl Radical; Lipid Peroxidation; Lipid Peroxides; Microwaves; Pressure; Sulfonic Acids; Vegetables; Water

Cerium (Ce) is a rare earth metal that is not known to have any biological role. Cerium oxide materials of several sizes and shapes have been developed in recent years as a scaffold for catalysts. Indeed even cerium oxide nanoparticles themselves have displayed catalytic activities and antioxidant properties in tissue culture and animal models. Because of ceria's ability to cycle between the +3 and +4 states at oxygen vacancy sites, we investigated whether cerium metal would catalyze a Fenton-like reaction with hydrogen peroxide. Indeed, cerium chloride did exhibit radical production in the presence of hydrogen peroxide, as assessed by relaxation of supercoiled plasmid DNA. Radical production in this reaction was also followed by production of radical cation of 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS). Radical scavengers and spin traps were capable of competing with ABTS for radicals produced in this cerium dependent Fenton-like reaction. Electron paramagnetic resonance experiments reveal both hydroxyl radical and superoxide anion in a reaction containing cerium and hydrogen peroxide. Based on these results we propose that cerium is capable of redox-cycling with peroxide to generate damaging oxygen radicals.

Topics: Benzothiazoles; Catalysis; Cerium; Chromans; DNA Damage; Electron Spin Resonance Spectroscopy; Free Radicals; Hydrogen Peroxide; Iron; Spin Trapping; Sulfonic Acids

The objective of this research was to re-evaluate the antioxidant effects of the prenylated flavonoids from Sophora flavescens via in vitro 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), peroxynitrite (ONOO(-)), and total reactive oxygen species (ROS) assays. In addition, a further examination of kuraridinol, kurarinol, and kurarinone, also isolated from S. flavescens, was carried out by the inhibition of tert-butylhydroperoxide (t-BHP)-induced intracellular ROS generation and t-BHP-induced activation of nuclear factor-kappaB (NF-kappaB). Upon re-examination of the ethyl acetate (EtOAc) soluble fraction of S. flavescens, two major prenylated chalcones, including kuraridin and kuraridinol, along with a minor prenylated flavonol, kushenol C, were isolated as good DPPH scavengers. This was in contrast to the prenylated flavanones, sophoraflavanone G and kurarinone, which were isolated from the methylene chloride (CH(2)Cl(2)) fraction of the same source. Five flavanones consisting of kushenol E, leachianone G, kurarinol, sophoraflavanone G, and kurarinone exhibited significant antioxidant potentials in the ABTS, ONOO(-), and total ROS assays; however, the prenylated chalcones and prenylated flavonol showed more potent scavenging/inhibitory activities than the prenylated flavanones. Therefore, the prenylated chalcones and prenylated flavonol, rather than the prenylated flavanones, may make important contributions toward the marked antioxidant capacities of S. flavescens. Furthermore, kuraridinol, kurarinol, and kurarinone showed significant inhibitory activities against intracellular ROS levels as well as NF-kappaB activation by t-BHP. Overall, the results indicate that S. flavescens and its prenylated flavonoids may possess good anti-inflammatory activity, which is implicated in their significant antioxidant activity.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Antioxidants; Benzothiazoles; Biphenyl Compounds; Cell Line; Chromans; Flavonoids; Genes, Reporter; Humans; Luciferases; Magnetic Resonance Spectroscopy; NF-kappa B; Peroxynitrous Acid; Picrates; Plant Extracts; Plant Roots; Prenylation; Rats; Reactive Oxygen Species; Sophora; Sulfonic Acids; tert-Butylhydroperoxide; Thiazoles

Thymoquinone (TQ) is the bioactive constituent of the volatile oil of Nigella sativa L. and has been shown to exert antioxidant antineoplastic and anti-inflammatory effects. During the study of its possible mechanism of action, we found that TQ reacts chemically (i.e. nonenzymatically) with glutathione (GSH), NADH and NADPH. A combination of liquid chromatography/UV-Vis spectrophotometry/Mass spectrometry analyses was used to identify the products of these reactions. The reaction that occur in physiological conditions indicates the formation of only two products, glutathionyl-dihydrothymoquinone after rapid reaction with GSH, and dihydrothymoquinone (DHTQ) after slow reaction time with NADH and NADPH. Measurement of the antioxidant activity of reduced compounds against organic radicals such as 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) also revealed a potential scavenging activity for glutathionyl-dihydrothymoquinone similar to that of DHTQ. Under our experimental conditions, TQ shows lower scavenging activities than glutathionyl-dihydrothymoquinone and DHTQ; it is very interesting to observe that the reduced compounds apparently show an antioxidant capacity equivalent to Trolox. The results indicate a possible intracellular nonenzymatic metabolic activation of TQ dependent on GSH, NADH or NADPH that may represent a "cellular switch" able to modulate cellular antioxidant defences.

Topics: Antioxidants; Benzoquinones; Benzothiazoles; Biphenyl Compounds; Chromans; Chromatography, High Pressure Liquid; Erythrocytes; Free Radical Scavengers; Glutathione; Humans; Hydrazines; Molecular Structure; NAD; NADP; Oxidation-Reduction; Picrates; Sulfonic Acids

The efficiencies of two traditional extraction methods used in Chinese medicine (the decoction method and the maceration method) were evaluated for the extraction of antioxidants from medicinal plants. A group of medicinal plants possessing nutritious and tonic functions were chosen as model plants. A commonly used extraction method was used as a reference method. The antioxidant capacities and total phenolic contents of the extracts were measured by ferric-reducing antioxidant power and Trolox equivalent antioxidant capacity assays as well as the Folin-Ciocalteu method, respectively. The results obtained indicated that the two traditional extraction methods could effectively extract antioxidants from medicinal plants. These extraction methods can be applied to the analysis and purification of antioxidants in plants, respectively. At home, people can use these methods to extract antioxidants from plants for consumption. In the food industry, these methods could be utilized to prepare crude extracts from plants containing antioxidants for use as food additives.

Topics: Antioxidants; Benzothiazoles; Chemistry Techniques, Analytical; Chromans; Drugs, Chinese Herbal; Ethanol; Ferrous Compounds; Iron; Medicine, Chinese Traditional; Methanol; Molybdenum; Oxidation-Reduction; Phenols; Plants, Medicinal; Reproducibility of Results; Sulfonic Acids; Triazines; Tungsten Compounds; Water

The effect of doxorubicin on oxidation of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) by lactoperoxidase and hydrogen peroxide has been investigated. It was found that: (1) oxidation of ABTS to its radical cation (ABTS*(+)) is inhibited by doxorubicin as evidenced by its induction of a lag period, duration of which depends on doxorubicin concentration; (2) the inhibition is due to doxorubicin hydroquinone reducing the ABTS*(+) radical (stoichiometry 1: 1.8); (3) concomitant with the ABTS*(+) reduction is oxidation of doxorubicin; only when the doxorubicin concentration decreases to a near zero level, net oxidation of ABTS could be detected; (4) oxidation of doxorubicin leads to its degradation to 3-methoxysalicylic acid and 3-methoxyphthalic acid; (5) the efficacy of doxorubicin to quench ABTS*(+) is similar to the efficacy of p-hydroquinone, glutathione and Trolox C. These observations support the assertion that under certain conditions doxorubicin can function as an antioxidant. They also suggest that interaction of doxorubicin with oxidants may lead to its oxidative degradation.

Topics: Antibiotics, Antineoplastic; Benzothiazoles; Chromans; Doxorubicin; Free Radicals; Glutathione; Hydrogen Peroxide; Hydroquinones; Lactoperoxidase; Oxidants; Oxidation-Reduction; Phthalic Acids; Salicylates; Sulfonic Acids; Thiazoles

Caffeic acid (3,4-dihydroxycinnamic acid) is among the major hydroxycinnamic acids present in wine; sinapic acid, which is a potent antioxidant. It has also been identified as one of the active antioxidant. In the present study, the antioxidant properties of the caffeic acid were evaluated by using different in vitro antioxidant assays such as 2-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging, 1,1-diphenyl-2-picryl-hydrazyl free radical (DPPH) scavenging, total antioxidant activity by ferric thiocyanate method, total reductive capability using the potassium ferricyanide reduction method, superoxide anion radical scavenging and metal chelating activities. alpha-Tocopherol, trolox, a water-soluble analogue of tocopherol, butylated hydroxyanisole (BHA), and butylated hydroxytoluene (BHT) were used as the reference antioxidant compounds. At the concentrations of 10 and 30 microg/mL, caffeic acid showed 68.2 and 75.8% inhibition on lipid peroxidation of linoleic acid emulsion, respectively. On the other hand, 20 microg/mL of standard antioxidant such as BHA, BHT, alpha-tocopherol and trolox indicated an inhibition of 74.4, 71.2, 54.7 and 20.1% on peroxidation of linoleic acid emulsion, respectively. In addition, caffeic acid is an effective ABTS(+) scavenging, DPPH scavenging, superoxide anion radical scavenging, total reducing power and metal chelating on ferrous ions activities.

Topics: alpha-Tocopherol; Antioxidants; Benzothiazoles; Biphenyl Compounds; Butylated Hydroxyanisole; Butylated Hydroxytoluene; Caffeic Acids; Chelating Agents; Chromans; Dose-Response Relationship, Drug; Emulsions; Ferricyanides; Ferrous Compounds; Free Radical Scavengers; Hydrazines; Iron; Linoleic Acid; Lipid Peroxidation; Picrates; Reducing Agents; Sulfonic Acids; Superoxides; Thiocyanates

Maytenus ilicifolia is an important plant with potential on cancer treatment and has been largely used in Brazil and other countries. We have evaluated the crude ethanolic extract of M. ilicifolia as a potential antioxidant source using an assay based on the bleaching of the radical monocation 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS(*+)) and by HOCl scavenger capacity. Trolox and uric acid were used as positive controls. The results indicated M. ilicifolia root bark as a great source of antioxidants based on its potential as scavenger of radicals.

Topics: Antioxidants; Benzothiazoles; Chromans; Ethanol; Free Radical Scavengers; Inhibitory Concentration 50; Maytenus; Plant Bark; Plant Extracts; Sulfonic Acids; Uric Acid

The aim of this study was to characterize the antioxidant activity of three ascorbic acid (AA) derivatives O-substituted at the C-2 position of AA: ascorbic acid 2-glucoside (AA-2G), ascorbic acid 2-phosphate (AA-2P), and ascorbic acid 2-sulfate (AA-2S). The radical-scavenging activities of these AA derivatives and some common low molecular-weight antioxidants such as uric acid or glutathione against 1,1-diphenyl-picrylhydrazyl (DPPH) radical, 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation (ABTS+), or galvinoxyl radical were kinetically and stoichiometrically evaluated under pH-controlled conditions. Those AA derivatives slowly and continuously reacted with DPPH radical and ABTS+, but not with galvinoxyl radical. They effectively reacted with DPPH radical under acidic conditions and with ABTS+ under neutral conditions. In contrast, AA immediately quenched all species of radicals tested at all pH values investigated. The reactivity of Trolox, a water-soluble vitamin E analogue, was comparable to that of AA in terms of kinetics and stoichiometrics. Uric acid and glutathione exhibited long-lasting radical-scavenging activity against these radicals under certain pH conditions. The radical-scavenging profiles of AA derivatives were closer to those of uric acid and glutathione rather than to that of AA. The number of radicals scavenged by one molecule of AA derivatives, uric acid, or glutathione was equal to or greater than that by AA or Trolox under the appropriate conditions. These data suggest the potential usage of AA derivatives as radical scavengers.

Topics: Ascorbic Acid; Benzhydryl Compounds; Benzothiazoles; Biphenyl Compounds; Chromans; Free Radical Scavengers; Glutathione; Hydrogen-Ion Concentration; Kinetics; Picrates; Sulfonic Acids; Uric Acid

The total antioxidant capacity (TAC) and phenolic composition of 139 Pinotage wines (2002 and 2003 vintages) were determined using the 2,2'-azino-di(3-ethylbenzo-thialozine-sulfonic acid) scavenging assay and high-performance liquid chromatography, respectively. The contribution of individually quantified phenolic compounds to the wine TAC was calculated using their concentrations and Trolox equivalent antioxidant capacity (TEAC) values. The TEAC values of quercetin-3-galactoside, isorhamnetin, and peonidin-3-glucoside are reported for the first time. Between 11 and 24% of the measured TAC of Pinotage wines was explained by the sum of the calculated contributions of their quantified phenolic compounds comprising monomeric phenolic compounds and procyanidin B1. Ultrafiltration was carried out to attempt separation of monomeric and polymeric phenolic compounds. Analysis of ultrafiltration permeates and retentates enabled estimation of the TAC contribution of large molecular weight (MW) unknown compounds (46%) (>50 kDa), including oligomeric and polymeric phenolic compounds and small MW unknown compounds (34%) (<50 kDa). Three mixtures, containing 12 phenolic compounds in typical concentrations expected in Pinotage wines, exhibited 16-23% synergistic antioxidant activity. This suggests that synergy between phenolic compounds does play a role in the wine TAC but that it does not explain the large discrepancy between measured and calculated TAC values.

Topics: Antioxidants; Benzothiazoles; Chromans; Free Radical Scavengers; Phenols; Sulfonic Acids; Ultrafiltration; Wine

In the present study, the pH-dependent free radical-scavenging activity of betanin in the Trolox equivalent antioxidant capacity (TEAC) assay was determined. It was found that at a pH > 4 betanin is about 1.5-2.0-fold more active than some anthocyanins considered very good free radical scavengers as determined in the TEAC assay. The increase in the TEAC values of betanin with increasing pH is discussed in terms of its calculated phenolic OH homolytic bond dissociation energy (BDE) and ionization potential (IP). The results suggest that the exceptionally high antioxidant activity of betanin is associated with an increasing of its H-donation and electron-donation ability when going from cationic state to mono-, di- and tri-deprotonated states present at basic solutions.

Topics: Benzothiazoles; Beta vulgaris; Betacyanins; Chromans; Free Radical Scavengers; Sulfonic Acids

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.

Topics: Antifungal Agents; Antioxidants; Benzothiazoles; Biphenyl Compounds; Chromans; Fungi; Gas Chromatography-Mass Spectrometry; Hydrazines; Oils, Volatile; Picrates; Plant Extracts; Plant Leaves; Sulfonic Acids; Teucrium; Tunisia

Minocycline is neuroprotective in animal models of a number of acute CNS injuries and neurodegenerative diseases. While anti-inflammatory and anti-apoptotic effects of minocycline have been characterized, the molecular basis for the neuroprotective effects of minocycline remains unclear. We report here that minocycline and a number of antioxidant compounds protect mixed neuronal cultures in an oxidative stress assay. To evaluate the role of minocycline's direct antioxidant properties in neuroprotection, we determined potencies for minocycline, other tetracycline antibiotics, and reference antioxidant compounds using a panel of in vitro radical scavenging assays. Data from in vitro rat brain homogenate lipid peroxidation and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assays show that minocycline, in contrast to tetracycline, is an effective antioxidant with radical scavenging potency similar to vitamin E. Our findings suggest that the direct antioxidant activity of minocycline may contribute to its neuroprotective effects in some cell-based assays and animal models of neuronal injury.

Topics: Animals; Antioxidants; Antipyrine; Benzothiazoles; Cell Death; Cells, Cultured; Cerebral Cortex; Chromans; Deoxyribose; Dizocilpine Maleate; Dose-Response Relationship, Drug; Drug Interactions; Edaravone; Embryo, Mammalian; Free Radical Scavengers; Free Radicals; Glutamic Acid; Inhibitory Concentration 50; L-Lactate Dehydrogenase; Lipid Peroxidation; Minocycline; Neurons; Neuroprotective Agents; Oxidative Stress; Phenethylamines; Rats; Rats, Sprague-Dawley; Sulfonic Acids; Vitamin E

In this paper, a fast strategy for determining the total antioxidant capacity of Chinese green tea extracts is developed. This strategy includes the use of experimental techniques, such as fast high-performance liquid chromatography (HPLC) on monolithic columns and a spectrophotometric approach to determine the total antioxidant capacity of the extracts. To extract the chemically relevant information from the obtained data, chemometrical approaches are used. Among them there are correlation optimized warping (COW) to align the chromatograms, robust principal component analysis (robust PCA) to detect outliers, and partial least squares (PLS) and uninformative variable elimination partial least squares (UVE-PLS) to construct a reliable multivariate regression model to predict the total antioxidant capacity from the fast chromatograms.

Topics: Antioxidants; Benzothiazoles; Camellia sinensis; Chromans; Chromatography, High Pressure Liquid; Least-Squares Analysis; Multivariate Analysis; Regression Analysis; Sulfonic Acids; Tea

Topics: Amidines; Antioxidants; Benzothiazoles; Chromans; Ferric Compounds; Fluorescent Dyes; Food Analysis; Fruit; Oxidation-Reduction; Phycoerythrin; Rosaceae; Sulfonic Acids; Triazines

The Trolox Equivalent Antioxidant Capacity (TEAC) assay is based on the scavenging of the 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical (ABTS(*)) converting it into a colorless product. The degree of decolorization induced by a compound is related to that induced by trolox, giving the TEAC value. The assay is frequently used for constructing structure activity relationships (SARs). HPLC analysis of the reaction mixture, obtained after scavenging of ABTS(*) by the flavonoid chrysin, shows that a product is formed that also reacts with ABTS(*). The product has a higher antioxidant capacity and reacts faster with ABTS(*) than the parent compound, chrysin. In contrast to the reaction product of chrysin, the reaction product of trolox, which is formed during scavenging of ABTS(*), i.e. trolox quinone, does not react with ABTS(*). The experiments show that the TEAC is the antioxidant capacity of the parent compound plus the potential antioxidant capacity of the reaction product(s). This means that the TEAC assay does not necessarily reflect the antioxidant effect of only one structure. This hampers the applicability of the assay for the construction of SARs and for ranking antioxidants.

Topics: Antioxidants; Benzothiazoles; Chromans; Chromatography, High Pressure Liquid; Flavonoids; Free Radical Scavengers; Quinones; Spectrophotometry, Ultraviolet; Structure-Activity Relationship; Sulfonic Acids

Topics: Antioxidants; Benzothiazoles; Chromans; Humans; Reactive Oxygen Species; Sulfonic Acids

The antioxidant and antimutagenic activities of the novel carboxyethyl derivatives of chitosan with three different degrees of substitution have been assayed in vitro in the unicellular flagellate Euglena gracilis subjected to the action of genotoxic agents acridine orange and ofloxacin. It has been demonstrated that chitosan derivatives exhibit concentration-dependent protective antigenotoxic activity against both mutagens. It is suggested that different mechanisms may be involved in its protective action--antioxidant activity in case of ofloxacin-induced DNA damage, as well as possible interaction with the cell membrane that prevents acridine orange from reaching the genetic compartments and subsequent damaging DNA through intercalative binding. Direct adsorption of acridine orange on chitosan derivatives was ruled out as a possible mechanism of protection on the basis of spectrophotometric measurements. Dependence of the antimutagenic properties of the studied chitosan derivatives on the degree of substitution was reversed in experiments involving acridine orange and ofloxacin, which also indicated different mechanisms of protection involved in these two cases.

Topics: Acridine Orange; Algorithms; Animals; Antimutagenic Agents; Antioxidants; Benzothiazoles; Carbohydrate Sequence; Chitosan; Chloroplasts; Chromans; DNA Damage; Esters; Euglena gracilis; Molecular Sequence Data; Mutagenicity Tests; Mutagens; Ofloxacin; Oxidants; Spectrophotometry, Ultraviolet; Sulfonic Acids

The 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation (ABTS(*)(+)) assay was adapted to a flow injection (FI) system to obtain a sensitive and rapid technique for the monitoring of antioxidant activity of pure compounds and complex matrixes, such as beverages and food extracts. The FI system includes a HPLC pump that flows the mobile phase (a solution of ABTS(*)(+) in ethanol) through a 20 microL loop injector, a single bead string reactor filled with acid-washed silanized beads, a delay coil and a photodiode array UV-visible detector. The technique was very sensitive, with limits of detection and of quantification of 4.14 and 9.29 micromol of Trolox/L, respectively, and demonstrated high repeatability and reproducibility. The proposed technique was then applied to the evaluation of the antioxidant activity of some pure compounds, demonstrating good agreement with published data obtained by the original spectrophotometric ABTS(*)(+) assay. Finally, the total antioxidant activity of 10 beverages was determined by both the proposed and the original method. The values ranged from 0.09 mmol L(-)(1) for cola to 49.24 mmol L(-)(1) for espresso coffee and did not result significantly different from those obtained by the original spectrophotometric ABTS(*)(+) assay (Student's paired t-test: t = 1.4074, p = 0.1929). In conclusion, the proposed FI technique seems suitable for the direct, rapid and reliable monitoring of total antioxidant activity of pure compounds and beverages and, due to the ability to operate in continuous, it allows the analysis of about 30 samples h(-)(1) making the assay particularly suitable for large screening of total antioxidant activity in food samples.

Topics: Antioxidants; Beer; Benzothiazoles; Beverages; Carbonated Beverages; Cations; Chromans; Citrus; Coffee; Flow Injection Analysis; Free Radicals; Fruit; Indicators and Reagents; Quality Control; Sensitivity and Specificity; Solutions; Spectrophotometry; Sulfonic Acids; Tea

A series of C4-substituted 1,4-dihydropyridines (DHP) with either secondary or tertiary nitrogen in the dihydropyridine ring were synthesized. All of these compounds together with some commercial DHP derivatives were tested for potential scavenger effects toward alkyl, alkylperoxyl radicals, and ABTS radical cation in aqueous media at pH 7.4. Kinetic rate constants were assessed either by UV/vis spectroscopy or GC/MS techniques. Tested compounds reacted faster toward alkylperoxyl radicals and ABTS radical cation than alkyl ones. N-Ethyl-substituted DHPs showed the lowest reactivity. Kinetic results were compared with either trolox or nisoldipine. Using deuterium kinetic isotope effect studies, we have proved that the hydrogen of the 1-position of the DHP ring is involved in the proposed mechanism. This fact is mostly noticeable in the case of alkyl radicals. In all cases, the respective pyridine derivative was detected as the main product of the reaction.

Topics: Benzothiazoles; Cations; Chromans; Dihydropyridines; Free Radical Scavengers; Free Radicals; Gas Chromatography-Mass Spectrometry; Hydrogen-Ion Concentration; Kinetics; Nisoldipine; Peroxides; Spectrophotometry, Ultraviolet; Sulfonic Acids; Vitamin E

Melatonin and classic antioxidants possess the capacity to scavenge ABTSb+ with IC50s of 4, 11, 15.5, 15.5, 17 and 21 microm for melatonin, glutathione, vitamin C, trolox, NADH and NADPH, respectively. In terms of scavenging ABTSb+, melatonin exhibits a different profile than that of the classic antioxidants. Classic antioxidants scavenge one or less ABTSb+, while each melatonin molecule can scavenge more than one ABTSb+, probably with a maximum of four. Classic antioxidants do not synergize when combined in terms of scavenging ABTSb+. However, a synergistic action is observed when melatonin is combined with any of the classic antioxidants. Cyclic voltammetry indicates that melatonin donates an electron at the potential of 715 mV. The scavenging mechanism of melatonin on ABTSb+ may involve multiple-electron donations via intermediates through a stepwise process. Intermediates including the melatoninyl cation radical, the melatoninyl neutral radical and cyclic 3-hydroxymelatonin (cyclic 3-OHM) and N1-acetyl-N2-formyl-5-methoxykynuramine (AFMK) seem to participate in these reactions. More interestingly, the pH of the solution dramatically modifies the ABTSb+ scavenging capacity of melatonin while pH changes have no measurable influence on the scavenging activity of classic antioxidants. An acidic pH markedly reduces the ABTSb+ scavenging capacity of melatonin while an increased pH promotes the interaction of melatonin and ABTSb+. The major melatonin metabolites that develop when melatonin interacts with ABTSb+ are cyclic 3-OHM and AFMK. Cyclic 3-OHM is the intermediate between melatonin and AFMK, and cyclic 3-OHM also has the ability to scavenge ABTSb+. Melatonin and the metabolites which are generated via the interaction of melatonin with ABTSb+, i.e. the melatoninyl cation radical, melatoninyl neutral radical and cyclic 3-OHM, all scavenge ABTSb+. This unique cascade action of melatonin, in terms of scavenging, increases its efficiency to neutralized ABTSb+; this contrasts with the effects of the classic antioxidants.

Topics: Antioxidants; Ascorbic Acid; Benzothiazoles; Cations; Chromans; Chromatography, High Pressure Liquid; Electrochemistry; Free Radical Scavengers; Free Radicals; Glutathione; Hydrogen-Ion Concentration; Kynuramine; Mechanics; Melatonin; NAD; NADP; Sulfonic Acids

The antioxidant activity of hydroxytyrosol, hydroxytyrosol acetate, oleuropein, 3,4-dihydroxyphenylelenolic acid (3,4-DHPEA-EA) and 3,4-dihydroxyphenylelenolic acid dialdehyde (3,4-DHPEA-EDA) towards oxidation initiated by 2,2'-azobis(2-amidinopropane) hydrochloride in a soybean phospholipid liposome system was studied. The antioxidant activity of these olive oil phenols was similar and the duration of the lag phase was almost twice that of alpha-tocopherol. Trolox, a water-soluble analogue of alpha-tocopherol, showed the worst antioxidant activity. However, oxidation before the end of the lag phase was inhibited less effectively by the olive oil phenols than by alpha-tocopherol and Trolox. Synergistic effects (11-20% increase in lag phase) were observed in the antioxidant activity of combinations of alpha-tocopherol with olive oil phenols both with and without ascorbic acid. Fluorescence anisotropy of probes and fluorescence quenching studies showed that the olive oil phenols did not penetrate into the membrane, but their effectiveness as antioxidants showed they were associated with the surface of the phospholipid bilayer.

Topics: alpha-Tocopherol; Amidines; Antioxidants; Ascorbic Acid; Benzothiazoles; Chromans; Fluorescence Polarization; Glycine max; Kinetics; Liposomes; Membrane Fluidity; Olive Oil; Oxidation-Reduction; Phenols; Plant Oils; Sulfonic Acids

To express the antioxidant capacity of plant foods in a more familiar and easily understood manner (equivalent to vitamin C mg/100 g), two stable radical species, ABTS(*)(-) and DPPH(*), commonly used for antioxidant activity measurements, were employed independently to evaluate their efficacies using apple polyphenolic extracts and seven polyphenolic standards including synthetic Trolox. Their antioxidant activities were expressed as vitamin C equivalent antioxidant capacity (VCEAC) in mg/100 g apple or mg/100 mL of the reference chemical compounds in 10 and 30 min using the ABTS(*)(-) and DPPH(*) scavenging assays, respectively. The antioxidant capacity of Gala apples and seven phenolic standards, determined by both ABTS(*)(-) and DPPH(*) scavenging assays, showed a dose-response of the first-order. Fresh Gala apples had a VCEAC of 205.4 +/- 5.6 mg/100 g using the ABTS assay, and the relative VCEACs of phenolic standards were as follows: gallic acid > quercetin > epicatechin > catechin > vitamin C > rutin > chlorogenic acid > Trolox. With the DPPH radical assay, the VCEAC of fresh Gala apples was 136.0 +/- 6.6 mg/100 g, and the relative VCEACs of seven phenolic standards were, in decreasing order, as follows: gallic acid > quercetin > epicatechin > catechin > or = vitamin C > Trolox > rutin > chlorogenic acid. Because the ABTS assay can be used in both organic and aqueous solvent systems, employs a specific absorbance at a wavelength remote from the visible region, and requires a short reaction time, it is a more desirable method than the DPPH assay. Therefore, it is recommended that antioxidant capacity be expressed as vitamin C mg/100 g equivalent (VCEAC) using the ABTS assay.

Topics: Antioxidants; Ascorbic Acid; Benzothiazoles; Biphenyl Compounds; Catechin; Chlorogenic Acid; Chromans; Free Radical Scavengers; Free Radicals; Gallic Acid; Kinetics; Malus; Oxidation-Reduction; Phenols; Picrates; Plant Extracts; Plants, Edible; Quercetin; Rutin; Sulfonic Acids

Peroxynitrite is implicated in numerous human diseases. Hence, there is considerable interest in potential therapeutic peroxynitrite scavengers. It has been claimed that uric acid is a powerful peroxynitrite scavenger. We previously observed that uric acid is a powerful inhibitor of tyrosine nitration induced by peroxynitrite, but fails to prevent alpha(1)-antiproteinase (alpha(1)-AP) inactivation induced by peroxynitrite. However, the reactivity of peroxynitrite is significantly modified by bicarbonate and this has not been considered in evaluating the scavenging activity of uric acid and other endogenous antioxidant compounds. In the presence of bicarbonate (25 mM), the ability of uric acid, ascorbate, Trolox, and GSH to inhibit peroxynitrite-mediated tyrosine and guanine nitration is decreased. Protection against peroxynitrite-mediated alpha(1)-AP inactivation is also decreased by ascorbate, Trolox, and GSH, but it is enhanced by uric acid. Bicarbonate also inhibits the ability of these compounds to prevent peroxynitrite-mediated ABTS radical cation formation. However, the abilities of these antioxidants to prevent peroxynitrite-mediated bleaching of pyrogallol red are enhanced by bicarbonate. These results show that physiologic concentrations of bicarbonate substantially modify the ability of uric acid to prevent peroxynitrite-mediated reactions. This study highlights the need to use several different assays in the presence of physiologically relevant concentrations of bicarbonate when assessing compounds for peroxynitrite scavenging, in order to avoid misleading results.

Topics: alpha 1-Antitrypsin; Antioxidants; Ascorbic Acid; Benzothiazoles; Bicarbonates; Chromans; Coloring Agents; Glutathione; Guanine; Humans; Indicators and Reagents; Peroxynitrous Acid; Pyrogallol; Reactive Nitrogen Species; Serine Proteinase Inhibitors; Sulfonic Acids; Tyrosine; Uric Acid

The use of N,N'-bis (2-hydroxybenzyl) ethylenediamine-N,N'-diacetic acid (HBED) for iron chelation therapy is currently being tested. Besides its affinity for iron, bioavailability, and efficacy in relieving iron overload, it is important to assess its anti- and/or pro-oxidant activity. To address these questions, the antioxidant/pro-oxidant effects of HBED in a cell-free solution and on cultured Chinese hamster V79 cells were studied using UV-VIS spectrophotometry, oximetry, spin trapping, and electron paramagnetic resonance (EPR) spectrometry. The results indicate that HBED facilitates Fe(II) oxidation but blocks O2(.-)-induced reduction of Fe(III) and consequently pre-empts production of .OH or hypervalent iron through the Haber-Weiss reaction cycle. The efficacy of HBED as a 1-electron donor (H-donation) was demonstrated by reduction of the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate)-derived nitrogen-centered radical cation (ABTS(.+)), accompanied by formation of a short-lived phenoxyl radical. HBED also provided cytoprotection against toxicity of H2O2 and t-BuOOH. Our results show that HBED can act both as a H-donating antioxidant and as an effective chelator lacking pro-oxidant capacity, thus substantiating its promising use in iron chelation therapy.

Topics: Animals; Antioxidants; Benzothiazoles; Cell Death; Cell Line; Cell Survival; Chromans; Cricetinae; Cricetulus; Cyclic N-Oxides; Cytoprotection; Edetic Acid; Electron Spin Resonance Spectroscopy; Hydrogen; Hydrogen Peroxide; Hydroxyl Radical; Iron Chelating Agents; Oxidation-Reduction; Oxygen; Phenols; Spectrophotometry; Spin Labels; Sulfonic Acids; Superoxides; tert-Butylhydroperoxide

Rapid scavenging of the model stable radical cation, ABTS(*+), has been applied to screen for the antioxidant activity of flavonoids. The reaction follows two distinct phases. For compounds with a monophenolic B-ring there is a rapid initial phase of reduction of ABTS(*+) within 0.1 s with no further change in the subsequent 2.9 s. In contrast, compounds with a catechol-containing B ring follow a fast initial scavenging phase with a slow secondary phase. Flavonoids with an unsubstituted B ring do not react within this time scale. The findings suggest that the structure of the B ring is the primary determinant of the antioxidant activity of flavonoids when studied through fast reaction kinetics.

Topics: Antioxidants; Benzopyrans; Benzothiazoles; Catechin; Catechols; Chromans; Coumaric Acids; Flavanones; Flavonoids; Flavonols; Free Radical Scavengers; Kinetics; Structure-Activity Relationship; Sulfonic Acids

Rates of free radical initiation were determined at 20 degrees C in 10 mM phosphate buffer (pH 7.4) in the systems metmyoglobin (methemoglobin)-H2O2 using 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) as the diammonium salt (ABTS). The catalytic activity of MetMb was 2-3-fold higher than that of MetHb. The process can be described by the Michaelis-Menten equation, from which effective values of Km and Vmax were calculated. Comparative kinetic studies on the inhibition of ABTS oxidation were carried out using Trolox, propylgallate (PG), polydisulfide of gallic acid (poly(DSG)), polydisulfide of (2-amino-4-nitrophenol) (poly(ADSNP)), and its conjugate with human serum albumin (HSA-poly(ADSNP)). The inhibitors were characterized by inhibition constants Ki and stoichiometric inhibition coefficients f (the number of radicals terminated by a single molecule of inhibitor). The minimum Ki and the maximum f values were obtained for poly(DSG), and in the system of MetHb-H2O2-ABTS they were 0.08 microM and 27.5, respectively. According to their antiradical activities, the inhibitors can be arranged as follows: poly(DSG) > poly(ADSNP) > PG > Trolox. PG, poly(DSG), poly (ADSNP), and its conjugate with HSA are suggested as "calibrators", i.e., inhibition standards for evaluation of antioxidant status of biological fluids in possible test systems based on the free radical-generating pair MetMb-H2O2 with ABTS as the acceptor of the active radicals.

Topics: Animals; Antioxidants; Benzothiazoles; Chromans; Flavonoids; Free Radicals; Humans; Hydrogen Peroxide; Kinetics; Methemoglobin; Metmyoglobin; Oxidation-Reduction; Phenols; Polymers; Polyphenols; Sulfonic Acids

A new cyclohexenone (1) and a new caffeoyl ester derivative (2), together with the known compounds (-)-isolariciresinol 3-alpha-O-beta-D-glucopyranoside (3), (+)-1-hydroxypinoresinol 1-O-beta-D-glucopyranoside (4), isoacteoside (5), luteolin 4'-O-beta-D-glucopyranoside (6), and indole-3-carboxylic acid (7), were isolated from the leaves of Bauhinia tarapotensis. The structures of these new compounds were determined by spectroscopic data analysis. The antioxidant activities of 1-7 were determined by measuring their free radical scavenging effects, using the 1,1-diphenyl-2-dipicrylhydrazyl free radical (DPPH) and Trolox equivalent antioxidant activity (TEAC) methods, and the coupled oxidation of beta-carotene and linoleic acid. Compounds 3-5 showed good activities in the DPPH and TEAC tests, while compounds 1 and 2 were active in the coupled oxidation of beta-carotene and linoleic acid bioassay.

Topics: Antioxidants; Benzothiazoles; Bepridil; Biphenyl Compounds; Caffeic Acids; Chromans; Chromatography, High Pressure Liquid; Cyclohexanones; Ecuador; Fabaceae; Flavonoids; Free Radical Scavengers; Glucosides; Indoles; Luteolin; Magnetic Resonance Spectroscopy; Molecular Structure; Phenols; Picrates; Plant Leaves; Plants, Medicinal; Sulfonic Acids

The effect of the pH on antioxidant properties of a series of hydroxyflavones was investigated. The pKa of the individual hydroxyl moieties in the hydroxyflavones was compared to computer-calculated deprotonation energies. This resulted in a quantitative structure activity relationship (QSAR), which enables the estimation of pKa values of individual hydroxyl moieties, also in hydroxyflavones for which these pKa values are not available. Comparison of the pKa values to the pH-dependent antioxidant profiles, determined by the TEAC assay, reveals that for various hydroxyflavones the pH-dependent behavior is related to hydroxyl moiety deprotonation, resulting in an increase of the antioxidant potential upon formation of the deprotonated forms. Comparison of these experimental results to computer calculated O-H bond dissociation energies (BDE) and ionization potentials (IP) of the nondeprotonated and the deprotonated forms of the various hydroxyflavones indicates that especially the parameter reflecting the ease of electron donation, i.e., the IP, and not the BDE, is greatly influenced by the deprotonation. Based on these results it is concluded that upon deprotonation the TEAC value increases (radical scavenging capacity increases) because electron-, not H*-, donation becomes easier. Taking into account that the mechanism of radical scavenging antioxidant activity of the neutral form of the hydroxyflavones is generally considered to be hydrogen atom donation, this implies than not only the ease of radical scavenging, but also the mechanism of antioxidant action changes upon hydroxyflavone deprotonation.

Topics: Antioxidants; Benzothiazoles; Chromans; Electron Transport; Flavonoids; Free Radical Scavengers; Hydrogen; Hydrogen-Ion Concentration; Hydroxyl Radical; Kinetics; Quantitative Structure-Activity Relationship; Sulfonic Acids; Thermodynamics

Scavenging of the ABTS (2,2'-azinobis[3-ethylbenzothiazoline-6-sulphonate])-derived nitrogen-centred radical cation (ABTS*+) was used to compare the total antioxidant activities of several seasonings used in Asian cooking. The results were expressed as Trolox equivalent antioxidant capacity (TEAC). The TEAC activities of dark soy sauces were found to be exceptionally high. In evaluating the TEAC of commercial products, attention must be paid to the addition of preservatives by manufacturers to the seasonings tested. Sodium benzoate (a preservative added to several seasonings) did not react significantly with ABTS*+, but the sulphite content of certain white wines may have led to an over-estimation of their TEAC.

Topics: Antioxidants; Asia; Benzothiazoles; Chromans; Cooking; Flavoring Agents; Food Preservatives; Glycine max; Hydrogen Peroxide; Kinetics; Sulfites; Sulfonic Acids; Wine

The antioxidant capacity of extracts of Crataegus oxyacantha, Hamamelis virginiana, Hydrastis canadensis, plants native to Europe and North America which have long been used in herbal medicine for the treatment of cardiac and circulatory functions, has been investigated. The total antioxidant potential conferred by all hydrogen donating antioxidants present in these extracts has been assessed by the ABTS assay and the relative order of antioxidant potential has been established. Gas chromatography coupled to mass spectrometry (GC-MS) has been used for the chemical identification of the antioxidant volatile compounds present in the extracts. The GC-MS data were related to the results obtained using the ABTS assay.

Topics: Antioxidants; Benzothiazoles; Chromans; Europe; Free Radicals; Gas Chromatography-Mass Spectrometry; Image Processing, Computer-Assisted; Indicators and Reagents; Magnoliopsida; North America; Plant Extracts; Plants, Medicinal; Rosales; Spectrophotometry, Ultraviolet; Sulfonic Acids

The 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulphonic acid) radical (ABTS(.+)) can be generated by the enzymatic system formed by hydrogen peroxide and horseradish peroxidase in an organic medium. The ABTS radical is easily generated in acidified ethanol medium in about 100 s with a stability of 1.7 x 10(-3) (-deltaabs/min) monitored at 730 nm. Other organic solvents, such as methanol or acetone, have lower radical generation times but the radical is less stable. The addition of Trolox or a lipophilic antioxidant such as alpha-tocopherol or beta-carotene produces a decrease in absorbance that can be used to estimate antioxidant capacity. Using a spectrophotometric end-point method and microplate-reader equipment, we have developed a method that estimates the antioxidant activity of different lipophilic vitamins. The use of Trolox as an antioxidant standard led to a limit of detection of 0.08 nmoles and limit of quantitation of 0.28 nmoles, while similar values were obtained for alpha-tocopherol and beta-carotene. The relative antioxidant activity values obtained by different antioxidants showed that alpha-tocopherol has a similar antioxidant potential to Trolox and that beta-carotene has 2.6 times the antioxidant potential of Trolox. In our opinion, this method can be useful for estimating the antioxidant activity in lipophilic samples and as a complement to other methods that measure antioxidant activity in aqueous media.

Topics: Antioxidants; Benzothiazoles; beta Carotene; Calibration; Chromans; Ethanol; Free Radicals; Horseradish Peroxidase; Indicators and Reagents; Reproducibility of Results; Sensitivity and Specificity; Solvents; Spectrophotometry; Sulfonic Acids; Vitamin E; Vitamins

Total reactive antioxidant potential (TRAP) of resinous exudates from Heliotropium species was evaluated by measuring the bleaching of stable free radicals. The antioxidant capacity of the resinous exudates in Trolox equivalents, evaluated from the bleaching of ABTS derived radical cations, ranged from 2.0 M (H. huascoense) to 5.2 M (H. stenophyllum), indicating a very high concentration of phenolic compounds. Considerably smaller values were obtained by measuring the bleaching of DPPH radicals. The ratio between the values obtained employing ABTS derived radicals and DPPH, ranged from 37 (H. megalanthum) to 4.5 (H. chenopodiaceum variety typica). The magnitude of the difference can be considered as an indication of the relative reactivity of the antioxidants present in the exudates. Similar ratios were observed when stoichiometric coefficients were evaluated for representative purified flavonoids obtained from the resinous exudates.

Topics: Antioxidants; Benzothiazoles; Bepridil; Biphenyl Compounds; Cations; Chromans; Flavonoids; Free Radicals; Phenols; Picrates; Plants; Resins, Plant; Sulfonic Acids

The influence of pH, intrinsic electron donating capacity, and intrinsic hydrogen atom donating capacity on the antioxidant potential of series of hydroxy and fluorine substituted 4-hydroxybenzoates was investigated experimentally and also on the basis of computer calculations. The pH-dependent behavior of the compounds in the TEAC assay revealed different antioxidant behavior of the nondissociated monoanionic form and the deprotonated dianionic form of the 4-hydroxybenzoates. Upon deprotonation the radical scavenging ability of the 4-hydroxybenzoates increases significantly. For mechanistic comparison a series of fluorobenzoates was synthesized and included in the studies. The fluorine substituents were shown to affect the proton and electron donating abilities of 4-hydroxybenzoate in the same way as hydroxyl substituents. In contrast, the fluorine substituents influenced the TEAC value and the hydrogen atom donating capacity of 4-hydroxybenzoate in a way different from the hydroxyl moieties. Comparison of these experimental data to computer-calculated characteristics indicates that the antioxidant behavior of the monoanionic forms of the 4-hydroxybenzoates is not determined by the tendency of the molecule to donate an electron, but by its ability to donate a hydrogen atom. Altogether, the results explain qualitatively and quantitatively how the number and position of OH moieties affect the antioxidant behavior of 4-hydroxybenzoates.

Topics: Antioxidants; Benzothiazoles; Chromans; Electrons; Fluorine; Free Radical Scavengers; Hydrogen; Hydrogen Bonding; Hydrogen-Ion Concentration; Hydroxylation; Indicators and Reagents; Parabens; Sulfonic Acids

A simple spectrophotometric method of determination of peroxyl radical-trapping capacity (PRTC) of body fluids and food products is proposed. In this method, decomposition of 2,2'-azobis(2-amidopropane) hydrochloride (ABAP) is the source of peroxyl and alkoxyl radicals which oxidize 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) to a green cation radical. Antioxidant present in a sample inhibit the reaction; the induction time of the reaction is proposed as a parameter enabling determination of antioxidant content. Standard assay conditions are: 20 mM ABAP and 150 microM ABTS in 0.1 M phosphate buffer, pH 7.0, at 37 degrees C; absorbance is monitored at 414 nm. A 10-min assay allows for determination of the induction time of appropriately diluted sample. As examples of application of this method, PRTC values of several types of beverages are reported.

Topics: Alcoholic Beverages; Amidines; Amino Acids; Antioxidants; Ascorbic Acid; Benzothiazoles; Chromans; Epinephrine; Free Radicals; Indicators and Reagents; Oxidation-Reduction; Peroxides; Plasma; Spectrophotometry; Sulfonic Acids; Time Factors

A range of catechins and oligomeric procyanidins was purified by high performance liquid chromatography (HPLC) from grape seed, apple skin, lentil and almond flesh. Catechins, galloylated epicatechin, glycosylated catechin, procyanidin dimers, galloylated dimers, trimer, and tetramer species were all identified, purified and quantified by HPLC, LC-MS and NMR. The antioxidant properties of these compounds were assessed using two methods: (a) inhibition of ascorbate/iron-induced peroxidation of phosphatidylcholine liposomes; (b) scavenging of the radical cation of 2,2'-azinobis(3-ethyl-benzothiazoline-6-sulphonate) (ABTS) relative to the water-soluble vitamin E analogue Trolox C (expressed as Trolox C equivalent antioxidant capacity, TEAC). Antioxidant activity in the lipid phase decreased with polymerisation in contrast with antioxidant action in the aqueous phase which increased from monomer to trimer and then decreased from trimer to tetramer. Galloylation of catechin and dimeric procyanidins decreased lipid phase and increased aqueous phase antioxidant activity. Glycosylation of catechin demonstrated decreased activity in both phases.

Topics: Anthocyanins; Antioxidants; Benzothiazoles; Biflavonoids; Catechin; Chromans; Dimerization; Fruit; Gallic Acid; Glycosylation; Inhibitory Concentration 50; Lipid Peroxidation; Liposomes; Phosphatidylcholines; Plants; Polymers; Proanthocyanidins; Seeds; Solubility; Sulfonic Acids

Topics: Antioxidants; Benzothiazoles; Blood Chemical Analysis; Chromans; Free Radicals; Humans; Indicators and Reagents; Spectrophotometry; Sulfonic Acids

A relatively simple and widely applied method for quantitating the total antioxidant capacity of body fluids and drug solutions based on the absorbance of the ABTS radical cation was evaluated. In this assay, the end-point is an antioxidant-induced decrease in absorbance at a fixed time. This decrease is used as an index of total antioxidant capacity. It is shown that Trolox, potassium cyanide and quercetin all decrease the absorbance of ABTS radical cations at a fixed time, but by different mechanisms. Trolox scavenges the ABTS radical, potassium cyanide inhibits radical formation, while quercetin acts by both mechanisms. Using this method antioxidant capacity may be overestimated, due to both a scavenger effect and an effect on the rate of ABTS oxidation. To distinguish between these effects, a post-addition assay was used in which the sample is added when the formation of radicals is stable. Using post- addition assay conditions enables discrimination between effects on radical scavenging and on the radical formation, two major mechanisms for antioxidant action. In extrapolating the results to an in vivo situation it should be questioned: (i) whether the peroxidase process does indeed mimic the process of radical formation in vivo, and (ii) whether the ABTS radicals do resemble the radical species involved in an in vivo situation. Results obtained in the ABTS radical-based methods should therefore be reviewed critically before the antioxidant capacity can be assessed.

Topics: Antioxidants; Benzothiazoles; Chromans; Cyanides; Free Radicals; Methods; Metmyoglobin; Oxidation-Reduction; Quercetin; Sulfonic Acids

The purpose of this study was to assess the relative antioxidant activities of a range of carotenes and xanthophylls through the extent of their abilities to scavenge the ABTS(.+) radical cation. The results show that the relative abilities of the carotenoids to scavenge the ABTS(.+) radical cation are influenced by the presence of functional groups with increasing polarities, such as carbonyl and hydroxyl groups, in the terminal rings, as well as by the number of conjugated double bonds.

Topics: Antioxidants; Benzothiazoles; beta Carotene; Carotenoids; Chromans; Cryptoxanthins; Free Radical Scavengers; Lutein; Structure-Activity Relationship; Sulfonic Acids; Xanthophylls

The 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical cation can be generated by incubation of ABTS and 2,2'-azo-bis(2- amidinopropane) at 45 degrees C. The ABTS radical cation is stable for several minutes at room temperature and reacts quantitatively and instantaneously with several antioxidants, such as Trolox, ascorbic acid, uric acid, cysteine, glutathione and bilirubin. In contrast, the ABTS radical cation reacts slowly with albumin. When serum is added to a solution of the ABTS radical cation, the bleaching of the radical follows biphasic kinetics, with a fast decay followed by a slow decay that takes place within several minutes. The fast decay is primarily due to uric acid, while the slow decay is related to the protein content of the sample. We propose that this procedure can provide an independent and simultaneous evaluation of the low molecular weight and protein antioxidants present in biological samples such as serum.

Topics: Antioxidants; Ascorbic Acid; Benzothiazoles; Bilirubin; Chromans; Cysteine; Glutathione; Indicators and Reagents; Sulfonic Acids; Temperature; Time Factors; Uric Acid

1. A new method has been developed for measuring the total antioxidant capacity of body fluids and drug solutions, based on the absorbance of the ABTS.+ radical cation. 2. An automated method for use on a centrifugal analyser, as well as a manual method, is described. 3. The procedure has been applied to physiological antioxidant compounds and radical-scavenging drugs, and an antioxidant ranking was established based on their reactivity relative to a 1.0 mmol/l Trolox standard. 4. The Trolox equivalent antioxidant capacity of plasma from an adult reference population has been measured, and the method optimized and validated. 5. The method has been applied to investigate the total plasma antioxidant capacity of neonates and how this may be compromised in prematurity.

Topics: Adult; Antioxidants; Benzothiazoles; Centrifugation; Chromans; Female; Humans; Infant, Newborn; Infant, Premature; Metmyoglobin; Reference Values; Spectrum Analysis; Sulfonic Acids

The reactions between Trolox C, a water-soluble vitamin E analogue, and several oxidizing free radicals including the hydroxyl radical and various peroxy radicals were examined by using the pulse-radiolysis technique. The results demonstrate that Trolox C may undergo rapid one-electron-transfer reactions as well as hydrogen-transfer processes; the resulting phenoxyl radical is shown to be relatively stable, in common with the phenoxyl radical derived from vitamin E. The reactions between the Trolox C phenoxyl radical and a variety of biologically relevant reducing compounds were examined by using both pulse radiolysis and e.s.r. The results demonstrate that the Trolox C phenoxyl radical is readily repaired by ascorbate (k = 8.3 x 10(6) dm3.mol-1.s-1) and certain thiols (k less than 10(5) dm3.mol-1.s-1) but not by urate, NADH or propyl gallate. Evidence from e.s.r. studies indicates that thiol-containing compounds may also enter into similar repair reactions with the alpha-tocopherol phenoxyl radical. Kinetic evidence is presented that suggests that Trolox C may 'repair' proteins that have been oxidized by free radicals.

Topics: Ascorbic Acid; Benzopyrans; Benzothiazoles; Binding, Competitive; Chromans; Electron Spin Resonance Spectroscopy; Electron Transport; Free Radicals; Hydrogen-Ion Concentration; Indicators and Reagents; Oxidation-Reduction; Pulse Radiolysis; Sulfhydryl Compounds; Sulfonic Acids