1-1-diphenyl-2-picrylhydrazyl and ethyl-acetate

Green tea contains a variety of biologically active constituents that are widely used in the pharmaceutical and food industries. Among them, simple catechins constitute a major group of compounds that is primarily responsible for the high biologic activity of green tea extracts. Therefore, the application of optimized extraction conditions may result in obtaining high value extracts. The main purpose of the study was to compare the content of polyphenols, mainly catechins, and the antioxidant activity of green tea extracts obtained by three different extraction methods: simple maceration, ultrasound extraction and accelerated solvent extraction using six various solvent systems. The quality of the extracts was evaluated by LC-ESI-Q-TOF-MS methodologies and spectrophotometric determinations. The obtained results revealed that catechins' extraction efficiency was identical for the three techniques studied. However, larger quantitative differences among the samples were observed when using different solvents. The total content of major catechins and gallic acid was within a very wide range of 10.2-842 mg/L. Ethyl acetate was by far the least effective extractant, regardless of the extraction technique used. After all, the solvent system composed of ethanol:water (1:1

Topics: Acetates; Antioxidants; Biphenyl Compounds; Catechin; Chromatography, Liquid; Ethanol; Extraction and Processing Industry; Gallic Acid; Mass Spectrometry; Picrates; Plant Extracts; Polyphenols; Principal Component Analysis; Solvents; Spectrophotometry; Tea; Ultrasonic Waves; Water

In the traditional system of medicine, leaves and stem bark of Euphorbia tithymaloides L. have been used for the treatment of asthma, persistent coughing, laryngitis, skin diseases and mouth ulcers. Some studies have reported the anti-inflammatory and antimicrobial activities of phytochemicals from the leaf; however, the analysis of essential oil and its antioxidant property is still unexplored.. This study evaluates the in vitro antioxidant potential of the essential oil and organic extracts from aerial parts of Euphorbia tithymaloides L.. Thirty one compounds representing 96.37% of total oil were detected by GC-MS, of which eugenol (22.52%), phenyl ethyl alcohol (14.63%), 3-pentanol (9.22%), caryophyllene oxide (7.73%), isoeugenol (7.32%), pentadecanol (5.14%), spathulenol (5.11%) and α-pinene (3.32%) were the major compounds. The oil and ethyl acetate extract displayed potent DPPH (IC50 = 13.67 and 17.59 µg/mL, respectively) and superoxide (IC50 = 21.83 and 42.34 µg/mL, respectively) radical-scavenging activities among all the tested samples. The oil and methanol extract also exhibited remarkable nitric oxide radical-scavenging activities (IC50 = 90.45 and 112.63 µg/mL, respectively) among other extracts. Furthermore, the methanol extract contained the highest amount of total phenolics as compared to other samples.. The results demonstrate that the oil and extracts of E. tithymaloides could serve as natural antioxidants for using in pharmaceutical or cosmetic industries.

Topics: Acetates; Antioxidants; Biphenyl Compounds; Chloroform; Euphorbia; Hexanes; Methanol; Oils, Volatile; Picrates; Plant Components, Aerial; Plant Extracts; Superoxides; Turkey

Background Zingiber officinale Roscoe has been used in traditional medicine for the treatment of neurological disorder. This study aimed to investigate the phenolic contents, antioxidant, acetylcholinesterase enzyme (AChE) inhibitory activities of different fraction of Z. officinale root grown in Vietnam. Methods The roots of Z. officinale are extracted with ethanol 96 % and fractionated with n-hexane, ethyl acetate (EtOAc) and butanol (BuOH) solvents. These fractions evaluated the antioxidant activity by 1,1-Diphenyl -2-picrylhydrazyl (DPPH) assay and AChE inhibitory activity by Ellman's colorimetric method. Results Our data showed that the total phenolic content of EtOAc fraction was highest equivalents to 35.2±1.4 mg quercetin/g of fraction. Our data also demonstrated that EtOAc fraction had the strongest antioxidant activity with IC50 was 8.89±1.37 µg/mL and AChE inhibitory activity with an IC50 value of 22.85±2.37 μg/mL in a dose-dependent manner, followed by BuOH fraction and the n-hexane fraction is the weakest. Detailed kinetic analysis indicated that EtOAc fraction was mixed inhibition type with Ki (representing the affinity of the enzyme and inhibitor) was 30.61±1.43 µg/mL. Conclusions Our results suggest that the EtOAc fraction of Z. officinale may be a promising source of AChE inhibitors for Alzheimer's disease.

Topics: Acetates; Acetylcholinesterase; Antioxidants; Biphenyl Compounds; Cholinesterase Inhibitors; Inhibitory Concentration 50; Phenols; Picrates; Plant Extracts; Vietnam; Zingiber officinale

Topics: 1-Butanol; Acetates; Anti-Bacterial Agents; Antioxidants; Bacteria; Biphenyl Compounds; Cistaceae; Flavonoids; Free Radical Scavengers; Microbial Sensitivity Tests; Phenols; Picrates; Plant Components, Aerial; Plant Extracts; Polyphenols; Solvents

Topics: 2-Acetylaminofluorene; Acetates; Alkaline Phosphatase; Animals; Antioxidants; Biphenyl Compounds; Chemical Fractionation; Chromatography, High Pressure Liquid; Free Radical Scavengers; Glutathione; Lipid Peroxidation; Liver; Male; Metabolic Detoxication, Phase I; Metabolic Detoxication, Phase II; Oxidative Stress; Phytochemicals; Picrates; Plant Extracts; Protective Agents; Pteris; Rats, Wistar; Transaminases; Tumor Suppressor Protein p53

The essential oil extracted from Desfontaine's rupturewort, Herniaria fontanesii J. Gay subsp. fontanesii growing wildly in Tunisia, was analyzed using GC and GC-MS techniques. The free radical scavenging capacity and total phenol contents of three crude extracts having different polarities (n-hexane, ethyl acetate and methanol) were examined. Thus, a total of 35 constituents were identified in the Desfontaine's rupturewort essential oil representing 89.8% of the whole constituents. The oil was dominated by hexadecanoic acid, caryophyllene oxide, terpin-4-ol, khusimone and trans-sabinene hydrate. The total phenolic contents ranged from 16.91 to 92.27 mg of gallic acid/g of dry weight and they were found to be significantly higher in methanol than in polar ethyl acetate and hexane extracts. Correlations were observed between the phenolic contents and the antioxidant properties. Thus, the antioxidant activity of the methanol extract was superior to that of all samples tested (IC50 = 0.21 ± 0.04 mg/mL).

Topics: Acetates; Bicyclic Monoterpenes; Biphenyl Compounds; Caryophyllaceae; Free Radical Scavengers; Gallic Acid; Gas Chromatography-Mass Spectrometry; Hexanes; Methanol; Monoterpenes; Oils, Volatile; Palmitic Acid; Picrates; Plant Components, Aerial; Plant Extracts; Polycyclic Sesquiterpenes; Sesquiterpenes; Solvents; Terpenes; Tunisia

An efficient method for the rapid extraction, separation and purification of bioactive lignans, arctiin and arctigenin, from Fructus arctii by microwave-assisted extraction coupled with high-speed countercurrent chromatography was developed. The optimal extraction conditions of arctiin and arctigenin were evaluated by orthogonal array. Arctigenin could be converted from arctiin by hydrochloric acid hydrolysis. The separations were performed at a preparative scale with two-phase solvents composed of ethyl acetate-ethanol-water (5 : 1 : 5, v/v/v) for arctiin, and n-hexane-ethyl acetate-ethanol-water (4 : 4 : 3 : 4, v/v/v/v) for arctigenin. From 500 mg of crude extract sample, 122.3 mg of arctiin and 45.7 mg of arctigenin were obtained with the purity of 98.46 and 96.57%, and the recovery of 94.3 and 81.6%, respectively. Their structures were determined by comparison with the high-performance liquid chromatography retention time of standard substance as well as UV, FT-IR, electrospray ion source (ESI)-MS, (1)H-NMR and (13)C-NMR spectrum. According to the antioxidant activity assay, arctigenin had stronger 1,1-diphenyl-2-picrylhydrazyl free radicals scavenging activity.

Topics: Acetates; Arctium; Biphenyl Compounds; Chromatography, High Pressure Liquid; Countercurrent Distribution; Ethanol; Free Radical Scavengers; Furans; Glucosides; Hexanes; Hydrochloric Acid; Hydrolysis; Lignans; Liquid-Liquid Extraction; Microwaves; Picrates; Plant Extracts; Solvents; Water

Context The present study deals with new biological properties of the wild edible Diplotaxis simplex (Viv.) Spreng (Brassicaceae). Objectives The current study evaluates the antioxidant, the anti-inflammatory and the anti-cancer properties of ethyl acetate and ethanol extracts from D. simplex flowers. Materials and methods The anti-proliferative activity of the extracts (10-70 μg/mL) was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) against human colon cancer cell line Caco-2. The anti-inflammatory potential was evaluated by the inhibitory effect of the extracts (1.5-7.5 mg/mL) on phospholipase A2 activity as well as on carrageenan-induced paw oedema in mice. Extracts (200 mg/kg) or indomethacin (50 mg/kg) as positive control were injected intraperitoneally for albino mice prior to the induction of the oedema by carrageenan. Antioxidant activities were investigated using various complementary methods. Results Flower extracts contained a high level of polyphenolics (17.10-52.70 mg GAE/g) and flavonoids (74.20-100.60 mg QE/g), which correlate with its appreciable antioxidant potential in β-carotene peroxidation (IC50 value: 12.50-27.10 μg/mL), DPPH(•) radical-scavenging (IC50 value: 0.20-0.40 mg/mL), Fe(3+ )reducing (EC50 value: 0.10-0.14 mg/mL) and Fe(2+ )chelating (IC50 value: 0.20-0.60 mg/mL) assays. These extracts were effective in inhibiting cancer cell growth (IC50 value: 62.0-63.25 μg/mL). Besides, the ethyl acetate extract inhibited phospholipase A2 activity (IC50 value: 2.97 mg/mL) and reduced the paw oedema in mice (from 0.38 ± 0.01 to 0.24 ± 0.01 cm), 4 h post-carrageenan challenge. Conclusion These data suggest that D. simplex may be useful as a candidate in the treatment of inflammation and the colon cancer.

Topics: Acetates; Animals; Anti-Infective Agents; Antineoplastic Agents, Phytogenic; Antioxidants; Biphenyl Compounds; Brassicaceae; Caco-2 Cells; Carrageenan; Cell Proliferation; Colonic Neoplasms; Disease Models, Animal; Dose-Response Relationship, Drug; Edema; Ethanol; Flowers; Humans; Inhibitory Concentration 50; Male; Mice; Phospholipase A2 Inhibitors; Phospholipases A2; Phytotherapy; Picrates; Plant Extracts; Plants, Medicinal; Solvents

Bergenia ciliata (locally known as Zakhm-e-hayat; wound healer) is commonly employed for wound healing, curing diarrhea and vomiting, fever, cough and pulmonary affections. Local community uses this plant as tea decoction with table salt. B. ciliata crude extract and its fractions were subjected to antibacterial, antioxidant effects as well as determination of total flavonoids and phenolics, DNA damage and anticancerous activities following standard protocols. Increased percentage inhibition of free radical in DPPH assay as well as elevated phenolic and flavonoid contents revealed antioxidant potential of this potent herb. Ethyl acetate and aqueous extracts showed IC(50) of 0.7 and 0.3 mg/ml respectively, against H157 cell line. Antibacterial analysis showed MIC 0.4-10mg/ml for crude extract and fractions. The results obtained conclude that extracts of B. ciliata contain remedial latent and can be used as possible source for drug development by pharmaceutical industries.

Topics: Acetates; Anti-Bacterial Agents; Antineoplastic Agents, Phytogenic; Antioxidants; Bacteria; Biphenyl Compounds; Cell Line, Tumor; Cell Survival; Chemical Fractionation; Dose-Response Relationship, Drug; Humans; Inhibitory Concentration 50; Microbial Sensitivity Tests; Neoplasms; Phytotherapy; Picrates; Plant Extracts; Plants, Medicinal; Rhizome; Saxifragaceae; Water

The antioxidant activities of ethyl acetate extraction of purple rice (EAEPR) were evaluated by various methods in vitro and in vivo. In in vitro antioxidant assays, EAEPR was found to have strong 2, 2-diphenyl-1-picrylhydrazyl radical scavenging activity hydroxyl radical, reducing power and metal-ion chelating activity. In in vivo antioxidant assays, mice were administered with EAEPR via gavage for 42 consecutive days. As a result, administration of EAEPR significantly enhanced the activities of glutathione peroxidase in serums and livers of mice. EAEPR could improve the lipid status, especially total cholesterol and low-density lipoprotein cholesterol levels. In addition, total phenolic content of EAEPR was 188.21mg/g. The main phenolic compounds in EAEPR analyzed by ultra-high performance liquid chromatography tandem mass spectrometry were determined as ferulic acid and quercetin. The contents of ferulic acid and quercetin in EAEPR were 14.21mg/g and 35.28mg/g, respectively. The Nrf2 expression was significantly elevated after administration of EAEPR.These results suggested that EAEPR had potent antioxidant activity and could be explored as a novel natural antioxidant.

Topics: Acetates; Animals; Antioxidants; Atherosclerosis; Biphenyl Compounds; Chelating Agents; Diet; Fatty Liver; Free Radical Scavengers; Gene Expression Regulation; Iron; Lipids; Liver; Male; Mice; Oryza; Oxidation-Reduction; Phenols; Picrates; Real-Time Polymerase Chain Reaction

The reactivity of tocopherols with 2,2-diphenyl-1-picrylhydrazyl (DPPH) was studied in model systems in order to establish a method for quantifying vitamin E in plant oils. The method was optimized with respect to solvent composition of the assay medium, which has a large influence on the course of reaction of tocopherols with DPPH. The rate of reaction of α-tocopherol with DPPH is higher than that of γ-tocopherol in both protic and aprotic solvents. In ethyl acetate, routinely applied for the analysis of antioxidant potential (AOP) of plant oils, reactions of tocopherols with DPPH are slower and concentration of tocopherols in the assay has a large influence on their molar reactivity. In 2-propanol, however, two electrons are exchanged for both α- and γ-tocopherols, independent of their concentration. 2-propanol is not toxic and is fully compatible with polypropylene labware. The chromatographically determined content of tocopherols and their molar reactivity in the DPPH assay reveal that only tocopherols contribute to the AOP of sunflower oil, whereas the contribution of tocopherols to the AOP of linseed oil is 75%. The DPPH assay in 2-propanol can be applied for rapid and cheap estimation of vitamin E content in plant oils where tocopherols are major antioxidants.

Topics: 2-Propanol; Acetates; alpha-Tocopherol; Antioxidants; Biological Assay; Biphenyl Compounds; Flax; gamma-Tocopherol; Kinetics; Oxidation-Reduction; Picrates; Plant Oils; Solvents; Sunflower Oil; Tocopherols; Vitamin E

Our study aimed to investigate the antidepressant-like effect of ethyl acetate extract of the flowers of Campsis grandiflora (EFCG) in a mice model of chronic unpredictable mild stress (CUMS).. HPLC-Q-TOF-MS was used to identify the chemical constituents of EFCG. The DPPH assay and ABTS radical-scavenging assay were performed to measure the antioxidant properties. The protective properties of EFCG against H2 O2 -induced oxidative damage were analysed in PC12 cells. The changes of behaviour profiles were investigated by using open-field test, sucrose preference test, forced swimming test (FST) and tail suspension test (TST). Brain tissue samples of mice were collected, and antioxidative measure levels were measured.. The result showed that EFCG had the most active anti-oxidative effect and the protective effect against H2 O2 oxidative injury in PC12 cells. Treatment with the EFCG significantly reduced the depressant-like severity and immobility period as compared with untreated CUMS mice in FST and TST. Moreover, EFCG significantly elevated the contents of superoxide dismutase, Glutathione Peroxidase and decreased the contents of Malonaldehyde (MDA) in mice brain.. Our study found first the antidepressant activity of the EFCG. The results suggested the therapeutic potential of EFCG for depressive disorder.

Topics: Acetates; Animals; Antidepressive Agents; Antioxidants; Behavior, Animal; Benzothiazoles; Bignoniaceae; Biphenyl Compounds; Brain; Chromatography, High Pressure Liquid; Disease Models, Animal; Flowers; Food Preferences; Male; Mice; Motor Activity; Oxidative Stress; PC12 Cells; Phytotherapy; Picrates; Plant Extracts; Plants, Medicinal; Rats; Solvents; Spectrometry, Mass, Electrospray Ionization; Stress, Psychological; Sulfonic Acids; Swimming

In this study, antioxidant activity, total phenolic and flavonoids content of four different fractions from the traditional Korean polyherbal medicine of Modified Bo-yang-Hwan-o-Tang (mBHT) was determined using spectrophotometric methods. Antioxidant activity of fractions was expressed as percentage of DPPH radicals inhibition and IC₅₀ values (μg/ml). Values in percentage ranged from 48.35 to 77.43%. The reducing powers of all the extracts were comparable with that of positive control sample of Butylated hydroxyl tolune (BHT) and ascorbic acid which was found to be dose dependent. Total phenolic content ranged from 106.83 ± 0.002 to 188.661 ± 0.002 mg/g, expressed as gallic acid equivalents. The total flavonoid contents varied from 28.44 ± 0.001 to 105.25 ± 0.001 mg/g, expressed as quarcetin equivalents. Ethyl acetate fractions of mBHT showed the highest phenolic (188.66 mg GAE/g) and flavonoids (105.25 mg QAE/g) contents and strong antioxidant activity. Total phenolics and flavonoid content of all the mBHT fractions were found reasonably correlated with IC₅₀ of DPPH (R²=0.980 and 0.932, respectively). The high contents of phenolic compounds indicated that these compounds responsible for antioxidant activity. Therefore, ethyl acetate fractions of mBHT can be regarded as promising candidates for natural plant sources of antioxidants.

Topics: Acetates; Antioxidants; Biphenyl Compounds; Chemical Fractionation; Flavonoids; Phenols; Picrates; Plant Extracts; Solvents

In this study, extracts were obtained from leaves of Pyrus communis L., Pyrus elaeagrifolia Pall., and Pyrus pyrifolia (Bum.) Nak. These extracts were tested for antiradical and antibacterial activity, as well as for the amount of total phenolic compounds, hydroquinone and arbutin. The antiradical activity was measured using 2,2-diphenyl-1-picrylhydrazyl radical and antibacterial activity with the disk diffusion method. The amount of phenolic compounds was determined using Folin Ciocalteu's phenol reagent, but the amount of hydroquinone and arbutin was measured with high performance liquid chromatography. The strongest antiradical activity was observed for ethyl acetate extract from leaves of P. communis L., and the lowest for the poorly soluble fraction (precipitate) from leaves of P. elaeagrifolia Pall. The highest number of antiradical units per gram of raw materials was noted for leaves of P. communis. The strongest antibacterial activity was measured for ethyl acetate extracts. The calculation of Spearman rank correlation coefficients indicated the existence of a positive correlation between contents of hydroquinone in extracts and their antibacterial activity for almost all investigated bacterial strains. The strains of fungi such as Candida albicans and Saccharomyces cerevisiae were completely resistant to the action of extracts.

Topics: Acetates; Anti-Bacterial Agents; Antioxidants; Arbutin; Bacillus subtilis; Biphenyl Compounds; Disk Diffusion Antimicrobial Tests; Escherichia coli; Helicobacter pylori; Hydroquinones; Picrates; Plant Extracts; Plant Leaves; Pseudomonas aeruginosa; Pyrus; Solvents; Species Specificity; Staphylococcus aureus

Chemical investigation of Codonopsis ovata resulted in the isolation and identification of β-sitosterol-3-O-glycoside, luteolin, apigenin, gentiacaulein, swertiaperenine, β-sitosterol, taraxeryl-3-acetate, and 3β-acetoxyoleanane-12-one. A rapid, precise, sensitive and validated HPTLC method for simultaneous quantification of these natural products (NPs) was developed on silica-gel 60F254 plate using ternary solvent system, n-hexane:ethyl acetate:formic acid (10.5:3.5:0.43, v/v/v). Markers were quantified after post chromatographic derivatization with cerric ammonium sulfate reagent. The method was validated for accuracy, precision, LOD, LOQ and all calibration curves showed a good linear relationship (r>0.9924) within test range. Precision was evaluated by intra- and inter-day tests with RSDs <2.59%, accuracy validation recovery 92.43-99.50% with RSDs <1.00%. Apigenin was found major component (natural abundance: 1.103%) and β-sitosterol the least (0.0263%). The NPs displayed antioxidant activity with luteolin exhibiting maximum effect at 1μg/mL concentration (75.9% for DPPH and 43.7% for ABTS) and others at 10 and 25μg/mL, suggesting thereby their apparent potential use for the prevention of free radical induced diseases or as an additive element to food and pharmaceutical industry.

Topics: Acetates; Ammonium Sulfate; Benzothiazoles; Biphenyl Compounds; Calibration; Chromatography, Thin Layer; Codonopsis; Formates; Free Radical Scavengers; Hexanes; Limit of Detection; Linear Models; Phytotherapy; Picrates; Plant Extracts; Plants, Medicinal; Reference Standards; Reproducibility of Results; Silica Gel; Solvents; Sulfonic Acids

In order to screen the Catalpa plant with high antioxidant activity and confirm the corresponding active fractions from Catalpa ovata G. Don, C. fargesii Bur., and C. bungei C. A. Mey., total flavonoid contents and antioxidant activities of the extracts/fractions of Catalpa plant leaves were determined. The determined total flavonoid content and antioxidant activity were used as assessment criteria. Those compounds with antioxidant activity were isolated with silica gel column chromatography and ODS column chromatography. Our results showed that the total flavonoid content in C. bungei C. A. Mey. (30.07 mg/g · DW) was the highest, followed by those in C. fargesii Bur. (25.55 mg/g · DW) and C. ovata G. Don (24.96 mg/g · DW). According to the determination results of total flavonoid content and antioxidant activity in 3 clones of leaves of C. bungei C. A. Mey., the total flavonoid content and antioxidant activity in crude extracts from C. bungei C. A. Mey. 6 (CA6) leaves were the highest. Moreover, the results showed that the total flavonoid content and antioxidant activities of ethyl acetate (EA) fraction in ethanol crude extracts in CA6 leaves were the highest, followed by n-butanol, petroleum ether (PE), and water fractions. Two flavonoid compounds with antioxidant activity were firstly isolated based on EA fraction. The two compounds were luteolin (1) and apigenin (2), respectively.

Topics: 1-Butanol; Acetates; Alkanes; Antioxidants; Apigenin; Bignoniaceae; Biphenyl Compounds; Ethanol; Hydroxyl Radical; Luteolin; Picrates; Plant Extracts; Plant Leaves; Solvents

Based on the ethnomedicinal uses and the effective outcomes of natural products in various diseases, this study was designed to evaluate Isodon rugosus as possible remedy in oxidative stress, alzheimer's and other neurodegenerative diseases. Acetylecholinestrase (AChE) and butyrylcholinesterase (BChE) inhibitory activities of crude methanolic extract (Ir.Cr), resultant fractions (n-hexane (Ir.Hex), chloroform (Ir.Cf), ethyl acetate (Ir.EtAc), aqueous (Ir.Aq)), flavonoids (Ir.Flv) and crude saponins (Ir.Sp) of I. rugosus were investigated using Ellman's spectrophotometric method. Antioxidant potential of I. rugosus was determined using DPPH, H2O2 and ABTS free radicals scavenging assays. Total phenolic and flavonoids contents of plant extracts were determined and expressed in mg GAE/g dry weight and mg RTE/g of dry sample respectively.. Among different fractions Ir.Flv and Ir.Cf exhibited highest inhibitory activity against AChE (87.44 ± 0.51, 83.73 ± 0.64%) and BChE (82.53 ± 0.71, 88.55 ± 0.77%) enzymes at 1 mg/ml with IC50 values of 45, 50 for AChE and 40, 70 μg/ml for BChE respectively. Activity of these fractions were comparable to galanthamine causing 96.00 ± 0.30 and 88.61 ± 0.43% inhibition of AChE and BChE at 1 mg/ml concentration with IC50 values of 20 and 47 μg/ml respectively. In antioxidant assays, Ir.Flv, Ir.Cf, and Ir.EtAc demonstrated highest radicals scavenging activities in DPPH and H2O2 assays which were comparable to ascorbic acid. Ir.Flv was found most potent with IC50 of 19 and 24 μg/ml against DPPH and H2O2 radicals respectively. Whereas antioxidant activates of plant samples against ABTS free radicals was moderate. Ir.Cf, Ir.EtAc and Ir.Cr showed high phenolic and flavonoid contents and concentrations of these compounds in different fractions correlated well to their antioxidant and anticholinestrase activities.. It may be inferred from the current investigations that the Ir.Sp, Ir.Flv and various fractions of I. rugosus are good sources of anticholinesterase and antioxidant compounds. Different fractions can be subjected to activity guided isolation of bioactive compounds effective in neurological disorders.

Topics: Acetates; Acetylcholinesterase; Antioxidants; Benzothiazoles; Biphenyl Compounds; Butyrylcholinesterase; Chloroform; Cholinesterase Inhibitors; Complex Mixtures; Flavonoids; Free Radical Scavengers; Free Radicals; Hexanes; Hydrogen Peroxide; Inhibitory Concentration 50; Isodon; Medicine, Traditional; Methanol; Oxidative Stress; Picrates; Plant Components, Aerial; Plant Extracts; Saponins; Spectrophotometry; Sulfonic Acids

Cactus (Opuntia spp) is widely cultivated as a vegetable, fruit, and forage crop and has been used in traditional medicine in American Indian, Mexican, and Korean cultures. Accumulative evidence from both in vitro and in vivo studies using cacti suggests their biological and pharmacological activities, such as their anti-cancer and anti-inflammatory roles in different cancer cells. In this study, the Opuntia humifusa stem (OHS) was extracted with different solvents and screened for radical scavenging activity using 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS˙(+)) and 1,1-diphenyl-2-picryl hydrazyl (DPPH). In addition, the total phenolic and flavonoid contents of each extract were analyzed using the Folin-Ciocalteu method and high performance liquid chromatography, respectively. Further, the cacti's bioactive fractions were evaluated for cell cytotoxicity and to understand their mechanism of action on human colon cancer (SW480) and breast cancer (MCF7) cells. An ethyl acetate (EtOAc) extract exhibited the highest cytotoxicity and resulted in an up-regulated expression of the pro-apoptotic protein Bax (bcl-2 associated X protein) and a down-regulated expression of the anti-apoptotic protein Bcl2 in both SW480 and MCF7 cells. The apoptosis was mediated through activation of caspase 8, 9, and 3/7 activities as well as PARP cleavage in SW480 cells, while the same extract activated only a caspase 9 activity in MCF7 cells. Furthermore, incubation of cells with the EtOAc extract down-regulated the expression of inflammatory molecules such as cyclooxygenase-2 (COX2) and inducible nitric oxide synthase (iNOS) in SW480 cells but not in MCF7 cells. Taken together, these results suggest that SW480 colon cancer cells are more susceptible to bioactive compounds present in OHS and may have potential in the prevention of cancer through modulation of apoptosis markers and inhibition of inflammatory pathways.

Topics: Acetates; Anti-Inflammatory Agents; Antioxidants; Apoptosis; bcl-2-Associated X Protein; Benzothiazoles; Biphenyl Compounds; Caspase 3; Caspase 7; Caspase 8; Caspase 9; Chromatography, High Pressure Liquid; Colon; Cyclooxygenase 2; Down-Regulation; Flavonoids; Free Radical Scavengers; Humans; MCF-7 Cells; Nitric Oxide Synthase Type II; Opuntia; Picrates; Plant Extracts; Sulfonic Acids; Up-Regulation

Research on natural products has gained a wide popularity due to the potential of discovering active compounds. The antioxidant properties contained in plants have been proposed as one of the mechanisms for the observed beneficial effect. Therefore, the present study investigated the antioxidant activity and total phenolic contents of various solvent extracts of Albizia procera leaves.. Antioxidant activity of the methanol extract and its derived fractions petroleum ether (APP), carbon tetrachloride (APC), dichloromethane (APD), ethyl acetate (APE), and residual aqueous fraction (APA) of the leaves of Albizia procera was performed by in vitro chemical analyses. Total phenolic content of the APM and other five fractions were also determined. APM and its derived fractions were also subjected to preliminary phytochemical screening test for various constituents.. Phytochemical screening revealed the presence of saponins, steroids, tannins, glycosides and flavonoids in the extracts. Amongst the extracts, APE showed the highest total phenolic content (449.18 ± 18.41mg of gallic acid equivalent/g of extract). In DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging test, the IC(50) value of APM, APP, APC, APD, APE and APA was 43.43, 63.60, 166.18, 41.15, 11.79, and 63.06 μg/mL, respectively. Therefore, among the APM and its derived fractions, APE showed the highest antioxidant activity which is comparable to that of standard ascorbic acid (AA) (IC(50) 10.12 μg/mL). The total antioxidant capacity was found to be varied in different fractions. The reducing activity on ferrous ion was ranked as APE > APD > APM > APA > APC.. The above evidences suggest that APE of A. procera leaf is a potential source of natural antioxidant and can be used to prevent diseases associated with free radicals.

Topics: Acetates; Antioxidants; Biphenyl Compounds; Free Radical Scavengers; Free Radicals; Methanol; Phenol; Picrates; Plant Extracts; Plant Leaves; Solvents

As the use of various anticancer drugs is associated with many undesirable side effects, there is an urgent need for the discovery of new, better, and specific anticancer compounds. Antioxidant and antiproliferative activities as well as effects on cell morphology were investigated for methanol (M), chloroform (C), ethyl acetate (E), and aqueous (A) extracts of Caulerpa peltata, Gelidiella acerosa, Padina gymnospora, and Sargassum wightii using 2,2-diphenyl-1-picrylhydrazyl radical-scavenging, ferrous ion chelation, and resazurin-based growth inhibition (in A549, HCT-15, MG-63, and PC-3 cell lines) assays. A general trend was the greater extraction of phenols and flavonoids by chloroform and ethyl acetate, which showed higher activity in many assays. These non-polar C and E extracts showed higher DPPH radical-scavenging and growth inhibitory activities in A549, HCT-15, and PC-3 cells. However, higher ferrous ion chelation (A extracts) and growth inhibition in MG-63 cells (M and A extracts) were seen for the polar extracts. Furthermore, P. gymnospora and C. peltata emerged as promising sources for antiproliferative agents that could be explored for their own activity and as leads for the development of other compounds.

Topics: Acetates; Analysis of Variance; Antineoplastic Agents; Biphenyl Compounds; Caulerpa; Cell Line, Tumor; Cell Proliferation; Chloroform; Drug Discovery; Flavonoids; Free Radical Scavengers; Humans; India; Iron Chelating Agents; Methanol; Phaeophyceae; Phenols; Picrates; Plant Extracts; Rhodophyta; Sargassum; Water

Hyaluronidases have been found as the target enzymes in the development of osteoarthritis (OA) disease. While there is still no curative treatment for this disease, recent studies on the treatment of OA were focused on the effectiveness of natural products which are expected to improve the symptoms with minimal side effects. The aim of this study was to screen selected Malaysian plants on their anti-hyaluronidase activity as well as to evaluate the active plant and its derived fractions on its potential anti-arthritic and antioxidant activities.. A total of 20 methanolic crude extracts (bark and leaf) from ten different plants were screened using a colorimetric hyaluronidase enzymatic assay. The active plant extract (Payena dasyphylla) was then studied for its hyaluronidase inhibitory activity in the interleukin-1β (IL-1β) stimulated human chondrocytes cell line (NHAC-kn) using zymography method. The Payena dasyphylla methanolic bark extract was then fractionated into several fractions in where the ethyl acetate (EA) fraction was evaluated for its inhibitory effects on the HYAL1 and HYAL2 gene expressions using reverse transcription-polymerase chain reaction (RT-PCR) technique. While the MMP-3 and MMP-13 protein expressions were evaluated using western blot method. The phenolic and flavonoid contents of the three fractions as well as the antioxidant property of the EA fraction were also evaluated.. Bark extract of Payena dasyphylla (100 μg/ml) showed the highest inhibitory activity against bovine testicular hyaluronidase with 91.63%. The plant extract also inhibited hyaluronidase expression in the cultured human chondrocyte cells in response to IL-1β (100 ng/ml). Similarly, treatment with Payena dasyphylla ethyl acetate (EA) fraction (100 μg/ml) inhibited the HYAL1 and HYAL2 mRNA gene expressions as well as MMP-3 and MMP-13 protein expression in a dose dependent manner. Payena dasyphylla EA fraction has demonstrated the highest amount of phenolic and flavonoid content with 168.62 ± 10.93 mg GAE/g and 95.96 ± 2.96 mg RE/g respectively as compared to water and hexane fractions. In addition, the Payena dasyphylla EA fraction showed strong antioxidant activity with IC₅₀ value of 11.64 ± 1.69 μg/mL.. These findings have shown that Payena dasyphylla might contained potential phenolic compounds that inhibiting the key enzyme in osteoarthritis development, which is the hyaluronidase enzyme through interruption of HYAL1 and HYAL1 gene expressions. The degradation of cartilage could also be inhibited by the plant through suppression of MMP-3 and MMP-13 protein expressions. We also reported that the inhibitory effect of Payena dasyphylla on hyaluronidase activity and expression might be due to its anti-oxidant property.

Topics: Acetates; Antioxidants; Biphenyl Compounds; Cell Line; Cells, Cultured; Chondrocytes; Flavonoids; Humans; Hyaluronoglucosaminidase; Interleukin-1beta; Metalloproteases; Methanol; Picrates; Plant Bark; Plant Extracts; Sapotaceae

The present study evaluated the antioxidant potential of Vernonia condensata Baker (Asteraceae). Dried and powdered leaves were exhaustively extracted with ethanol by static maceration followed by partition to obtain the hexane, dichloromethane, ethyl acetate, and butanol fractions. Total phenols and flavonoids contents were determined through spectrophotometry and flavonoids were identified by HPLC-DAD system. The antioxidant activity was assessed by DPPH radical scavenging activity, TLC-bioautography, reducing power of Fe(+3), phosphomolybdenum, and TBA assays. The total phenolic content and total flavonoids ranged from 0.19 to 23.11 g/100 g and from 0.13 to 4.10 g/100 g, respectively. The flavonoids apigenin and luteolin were identified in the ethyl acetate fraction. The IC50 of DPPH assay varied from 4.28 to 75.10 µg/mL and TLC-bioautography detected the antioxidant compounds. The reducing power of Fe(+3) was 19.98 to 336.48  μg/mL, while the reaction with phosphomolybdenum ranged from 13.54% to 32.63% and 56.02% to 135.00% considering ascorbic acid and rutin as reference, respectively. At 30 mg/mL, the ethanolic extract and fractions revealed significant effect against lipid peroxidation. All these data sustain that V. condensata is an important and promising source of bioactive substances with antioxidant activity.

Topics: Acetates; Antioxidants; Apigenin; Biphenyl Compounds; Chromatography, High Pressure Liquid; Flavonoids; Free Radical Scavengers; Luteolin; Malondialdehyde; Molybdenum; Oxidation-Reduction; Phenols; Phosphoric Acids; Picrates; Plant Extracts; Spectrophotometry, Ultraviolet; Thiobarbituric Acid Reactive Substances; Vernonia

Eugenia polyantha bark extracts were found to have potential antioxidative activities. This study is an effort to investigate the antioxidative compounds in E. polyantha. In vitro antioxidatve assay were used as guided tools for the isolation of antioxidative compounds. The methanol-water extracts showed the highest level of free radical-scavenging activity (ED50) = 180 microg mL(-1) and protection from beta-carotene bleaching (8.7 microg mL(-1)). The methanol-water (1:1) extracts exhibited strong DPPH scavenging activity and protection against beta carotene bleaching and was subjected to repeated silica gel column chromatography. The n-butanol, acetone and ethyl acetate solubles exhibited the highest antioxidative activities, derivatization was conducted to the isolated antioxidative compounds prior to identification. Catechin, gallic acid and rutin were isolated from those solubles as active compounds present in the Eugenia polyantha bark.

Topics: 1-Butanol; Acetates; Acetone; Antioxidants; beta Carotene; Biological Assay; Biphenyl Compounds; Chemical Fractionation; Chromatography; Methanol; Oxidation-Reduction; Phytotherapy; Picrates; Plant Bark; Plants, Medicinal; Silica Gel; Solvents; Syzygium; Water

Tridax procumbens L., Asteraceae, has been extensively used for various ailments in the Ayurvedic system of medicine. Previous studies have revealed remarkable phytoconstituents from Tridax procumbens L. with significant antioxidant activity. The aim of the present study is to measure the anti-DPPH activity of the purified isolated compounds from n butanol soluble part and ethyl acetate soluble part of successive methanolic extract of Tridax procumbens L. We thus quantified the total phenolic and total flavonoids in different purified isolated compounds, the whole of the tests were evaluated with a sample cone. of 100 microg mL(-1) and were determined spectrophotometrically using Folin-ciocaltue and AlCl3 reagents, respectively. DPPH (1,1-diphenyl, 2-picryl hydrazyl) assay was used to determine the in vitro antioxidant activity of different isolated compounds. Isolated compounds, one from ethyl acetate soluble part (EF-I) and one from n butanol soluble part (BF-II) were reported to possess a significant anti DPPH activity with lowest IC50 values 67.26 and 80.90 microg mL(-1), respectively while comparable to standard ascorbic acid with IC50 value of 59.62 microg mL(-1), due to the high concentration of phenols 146.4 microg mL(-1) from EF-I and 142.2 microg mL(-1) from BF-II and flavonoids 48 and 42.5 microg mL(-1) found in EF-I and BF-II isolated compounds, respectively.

Topics: 1-Butanol; Acetates; Antioxidants; Asteraceae; Biphenyl Compounds; Flavonoids; Methanol; Phenols; Phytotherapy; Picrates; Plant Extracts; Plant Weeds; Plants, Medicinal; Solvents; Spectrophotometry

Considering the growing evidence of the presence of antioxidant compounds in plant extracts, the objectives of this study were to identify antioxidant compounds in Lindera obtusiloba Blume (Lauraceae) and to evaluate their antimelanogenic activities on B16F10 melanoma cells. Organic solvent fractions were separated from L. obtusiloba extracts (LOE). The ethyl acetate fraction (LOE-E) was significantly active against oxidative damage induced by tert-butyl hydroperoxide in primary rat hepatocytes. Two single purified compounds, quercitrin (quercetin-3-O-α-L-rhamnopyranoside) and afzelin (kaempferol-3-O-α-L-rhamnoside), were identified by HPLC and NMR. These compounds were evaluated for antioxidant activities by 1,1-diphenyl 2-picrylhydrazyl (DPPH) radical scavenging assay and ferric reducing antioxidant power (FRAP) assay, and for their antimelanogenic activities by tyrosinase inhibitory assay melanin formation inhibition assay and Western bolt analysis for the signaling pathway. The significant effects of quercitrin on antioxidant and antimelanogenic activities, and signal modulation of ERK and MITF in B16F10 melanoma cells were observed. This is the first report to identify quercitrin in L. obtusiloba and its whitening effect.

Topics: Acetates; Animals; Antioxidants; Biphenyl Compounds; Cell Line, Tumor; Extracellular Signal-Regulated MAP Kinases; Gene Expression; Hepatocytes; Lindera; Male; Mannosides; Melanins; Melanoma, Experimental; Microphthalmia-Associated Transcription Factor; Monophenol Monooxygenase; Picrates; Plant Extracts; Primary Cell Culture; Proanthocyanidins; Quercetin; Rats; Rats, Sprague-Dawley; Signal Transduction; tert-Butylhydroperoxide

The antioxidant efficiency of dry extracts from inflorescences and/or leaves of seven Sorbus species was studied using four in vitro tests of SET (single electron transfer) and HAT-type (hydrogen atom transfer) mechanisms. The 70% methanol extracts and its diethyl ether, ethyl acetate, n-butanol and water fractions were tested in parallel with the phenolic standards, e.g., caffeic acid, quercetin, BHA, BHT, and Trolox. The SET-type activity of the extracts depended primarily on the extraction solvent. The most valuable extracts were n-butanol and ethyl acetate ones, which activity was high in the DPPH (EC(50) = 3.2-5.2 μg/mL), TEAC (2.8-4.0 mmol Trolox/g), and FRAP (9.8-13.7 mmol Fe2+/g) tests, and strongly correlated with the total phenolic levels (39.6-58.2% of gallic acid equivalents). The HPLC-PDA analysis of the extracts led to the identification of chlorogenic acid, isoquercitrin, hyperoside, rutin, quercetin 3-O-sophoroside, and sexangularetin 3-O-β-D-glucopyranoside as the main components. Apart from flavonoids and hydroxycinnamic acids, proanthocyanidins have also a significant impact on the SET-type activity. The HAT-reactivity of the extracts in the linoleic acid peroxidation test (IC(50) = 36.9-228.3 μg/mL) depended more strongly on the plant tissue than on the extraction solvent, and its correlation with the phenolic content was weak. Both SET and HAT-type activity of the most potent Sorbus extracts was comparable with the activity of the standards, indicating their great potential as effective sources for health products.

Topics: Acetates; Benzothiazoles; Biphenyl Compounds; Butanols; Chloroform; Chromatography, High Pressure Liquid; Coumaric Acids; Ether; Free Radical Scavengers; Humans; Lipid Peroxidation; Methanol; Oxidation-Reduction; Phenols; Picrates; Plant Extracts; Reference Standards; Solid Phase Extraction; Solvents; Sorbus; Sulfonic Acids

Bryophyllum pinnatum (Lank.) Oken (Crassulaceae) is a perennial succulent herb widely used in traditional medicine to treat many ailments. Its wide range of uses in folk medicine justifies its being called "life plant" or "resurrection plant", prompting researchers' interest. We describe here the isolation and structure elucidation of antimicrobial and/or antioxidant components from the EtOAc extract of B. pinnatum.. The methanol extract displayed both antimicrobial activities with minimum inhibitory concentration (MIC) values ranging from 32 to 512 μg/ml and antioxidant property with an IC50 value of 52.48 μg/ml. Its partition enhanced the antimicrobial activity in EtOAc extract (MIC = 16-128 μg/ml) and reduced it in hexane extract (MIC = 256-1024 μg/ml). In addition, this process reduced the antioxidant activity in EtOAc and hexane extracts with IC50 values of 78.11 and 90.04 μg/ml respectively. Fractionation of EtOAc extract gave seven kaempferol rhamnosides, including; kaempferitrin (1), kaempferol 3-O-α-L-(2-acetyl)rhamnopyranoside-7-O-α-L-rhamnopyranoside (2), kaempferol 3-O-α-L-(3-acetyl)rhamnopyranoside-7-O-α-L-rhamnopyranoside (3), kaempferol 3-O-α-L-(4-acetyl)rhamnopyranoside-7-O-α-L-rhamnopyranoside (4), kaempferol 3-O-α-D- glucopyranoside-7-O-α-L-rhamnopyranoside (5), afzelin (6) and α-rhamnoisorobin (7). All these compounds, except 6 were isolated from this plant for the first time. Compound 7 was the most active, with MIC values ranging from 1 to 2 μg/ml and its antioxidant activity (IC50 = 0.71 μg/ml) was higher than that of the reference drug (IC50 = 0.96 μg/ml).. These findings demonstrate that Bryophyllum pinnatum and some of its isolated compounds have interesting antimicrobial and antioxidant properties, and therefore confirming the traditional use of B. pinnatum in the treatment of infectious and free radical damages.

Topics: Acetates; Anti-Infective Agents; Antioxidants; Bacteria; Biphenyl Compounds; Candida; Glycosides; Hexanes; Inhibitory Concentration 50; Kaempferols; Kalanchoe; Methanol; Microbial Sensitivity Tests; Molecular Structure; Picrates; Plant Extracts

The aim of this study was to investigate the antioxidant and free-radical scavenging activity of extract and fractions from various parts of Elsholtzia ciliata. The inflorescences, leaves, stems and roots of E. ciliata were extracted separately and two phenolic component enrichment methods: ethyl acetate-water liquid-liquid extraction and macroporous resin adsorption-desorption, were adopted in this study. The antioxidant activities of water extracts and fractions of E. ciliata were examined using different assay model systems in vitro. The fraction root E (purified by HPD300 macroporous resin) exhibited the highest total phenolics content (497.2 ± 24.9 mg GAE/g), accompanied with the highest antioxidant activity against various antioxidant systems in vitro compared to other fractions. On the basis of the results obtained, E. ciliata extracts can be used potentially as a ready accessible and valuable bioactive source of natural antioxidants.

Topics: Acetates; Antioxidants; Biphenyl Compounds; Lamiaceae; Liquid-Liquid Extraction; Phenols; Picrates; Plant Components, Aerial; Plant Extracts; Plant Roots; Solid Phase Extraction; Water

A solvent system consisting of ethyl acetate, ethyl alcohol and water, in the volume ratio of 4.5:4.5:1, was developed and used to extract, at room temperature, betulin from white birch bark and antioxidants from spices (rosemary, thyme, sage, and oregano) and white oak chips. In addition, under reflux conditions, trimyristin was extracted from nutmeg using the same solvent system, and eugenol from olives was extracted using a mixture of salt water and ethyl acetate. The protocol demonstrates the use of water in organic solvents to extract natural products from plants. Measurement of the free-radical scavenging activity using by 2,2-diphenyl-1-picrylhydrazyl (DPPH) indicated that the extraction of plant material using ethyl acetate, ethyl alcohol and water (4.5:4.5:1, v/v/v) was exhaustive when carried out at room temperature for 96 h.

Topics: Abietanes; Acetates; Betula; Biphenyl Compounds; Ethanol; Eugenol; Free Radical Scavengers; Free Radicals; Liquid-Liquid Extraction; Myristica; Origanum; Picrates; Plant Bark; Plant Extracts; Quercus; Rosmarinus; Salvia officinalis; Solvents; Thymus Plant; Triglycerides; Triterpenes; Water

To evaluate the antioxidant potential of different extract/fractions of Anthocephalus cadamba (A. cadamba) (Roxb.) Miq. (Rubiaceae) and study the tentative identification of their active constituents.. The extract/fractions were screened for antioxidant activity using various in vitro assays viz. DPPH assay, ABTS assay, superoxide anion radical scavenging assay, reducing power assay and plasmid DNA nicking assay. Total phenolic content of extract/fractions was determined by colorimetric method. An ultra-performance LC-electrospray-quadrupole-time of flight mass spectrometry method was used to analyse the active constituents of extract/fractions of A. cadamba.. The ethyl acetate fraction was found to be most active fraction in all the assays as compared to other extract/fractions. The IC(50) value of ethyl acetate fraction (ETAC fraction) was 21.24 μg/mL, 1.12 μg/mL, 9.68 μg/mL and 57.81 μg/mL in DPPH assay, ABTS assay, reducing power assay and superoxide scavenging assay respectively. All the extract/fractions also showed the potential to protect the plasmid DNA (pBR322) against the attack of hydroxyl radicals generated by Fentońs reagent. The bioactive compounds were identified by UPLC-ESI-QTOF-MS, by comparing the mass and λ(max) with literature values.. The potential of the extract/fractions to scavenge different free radicals in different systems indicated that they may be useful therapeutic agents for treating radical-related pathologic damage.

Topics: Acetates; Antioxidants; Biphenyl Compounds; Chromatography, High Pressure Liquid; Colorimetry; DNA Damage; DNA, Circular; Oxidation-Reduction; Phenols; Picrates; Plant Extracts; Plant Leaves; Plasmids; Regression Analysis; Rubiaceae; Spectrometry, Mass, Electrospray Ionization

Selaginella sinensis (Selaginellaceae) is used extensively in traditional Chinese medicine (TCM) for the treatment of many kinds of chronic diseases. In this study, fractionation of the methanol extract of S. sinensis by different polarity solvents indicated the ethyl acetate fraction exhibited an potent 1,1-diphenyl-2-picryhydrazyl (DPPH) radical scavenging activity with the IC(50) value of 44.9 μM. In order to evaluate the scientific basis, antioxidant peaks were firstly screened using DPPH spiking test through high performance liquid chromatography (DPPH-HPLC). Under the target-guidance of DPPH-HPLC experiment, two flavonoids and six biflavonoids, quercetin (1), apigenin (2), amentoflavone (3), robustaflavone (4), 2,3-dihydroamentaflavone (5), hinokiflavone (6), 4'-O-methyl-robustaflavone (7) and ginkgetin (8) were separated by high-speed counter-current chromatography (HSCCC) method using n-hexane-ethyl acetate-methanol-water (8:8:9:7) as the solvent system with purities 98.2%, 97.6%, 99.4%, 92.3%, 98.5%, 98.9% and 99.6%, respectively. The structures were identified by electrospray ionization mass spectrometry (ESI-MS) and nuclear magnetic resonance (NMR) analysis. Antioxidant activity of eight isolated compounds was assessed by the radical scavenging effect on DPPH radical, compound 1 showed strongest antioxidant activities with IC(50) values of 3.2 μM, while compounds 2-8 showed weak antioxidant activities. This is the first report on simultaneous separation of eight antioxidant compounds from S. sinensis by HSCCC, moreover, apigenin and 4'-O-methyl-robustaflavone were first identified from this plant. Results of the present study indicated that the combinative method using DPPH-HPLC and HSCCC could be widely applied for rapid screening and isolating of antioxidants from complex TCM extract.

Topics: Acetates; Antioxidants; Biphenyl Compounds; Chemical Fractionation; Chromatography, High Pressure Liquid; Countercurrent Distribution; Flavonoids; Nuclear Magnetic Resonance, Biomolecular; Picrates; Plant Extracts; Selaginellaceae; Solvents

Three extracts of different polarities of Glycyrrhiza glabra L. leaves were characterized and evaluated for their antioxidant, anti-genotoxic and anti-inflammatory activity. In total, thirty components have been identified and quantified through the use of liquid chromatography (LC) with ultraviolet-visible diode-array-detector (UV-vis-DAD) and mass spectrometry (MS). The main components belong to the polyphenols family, being flavonoid and dihydrostilbene derivatives. The extracts have been investigated for their antioxidant, anti-genotoxic and anti-inflammatory activities, which are fundamental requirements of efficacious chemo-preventive agents. The ethyl acetate extract proved to be the most valuable, evidently for the conspicuous presence of several polyphenols, namely flavonoids and dihydrostilbenes.

Topics: Acetates; Anti-Inflammatory Agents; Antioxidants; Biphenyl Compounds; Colorimetry; Drug Evaluation, Preclinical; Drugs, Chinese Herbal; Glycyrrhiza; Hexanes; Methanol; Molybdenum; Mutagenicity Tests; Picrates; Plant Extracts; Plant Leaves; Tungsten Compounds

Methanolic extract of Jasminum mesnyi Hance leaves having antidiabetic activity was subjected to fractionation to obtain antioxidant and antihyperglycemic rich fraction. Different concentrations of ethyl acetate and n-butanol fractions were subjected to antioxidant assay by DPPH method, nitric oxide scavenging activity and reducing power assay. The fractions showed dose dependent free radical scavenging property in all the models. IC50 values for ethyl acetate and n-butanol fractions were 153.45 +/- 6.65 and 6.22 +/- 0.25 microg/ml, respectively, as compared to L-ascorbic acid and rutin (as standards; IC50 values 6.54 +/- 0.24 and 5.43 +/- 0.21 microg/ml, respectively) in DPPH model. In nitric oxide scavenging activity, IC50 values were 141.54 +/- 9.95 microg/ml, 35.12 +/- 1.58 microg/ml, 21.06 +/- 0.95 microg/ml and 29.93 +/- 0.32 microg/ml for ethyl acetate, n-butanol fractions, L-ascorbic acid and rutin, respectively. n-Butanol fraction showed a good reducing potential and better free radical scavenging activity as compared to ethyl acetate fraction. Potent antioxidant n-butanol fraction showed better oral glucose tolerance test (antihyperglycemic) at par with metformin (standard drug), n-Butanol fraction contained secoiridoid glycosides which might be responsible for both antioxidant and antihyperglycemic activity.

Topics: 1-Butanol; Acetates; Animals; Antioxidants; Biphenyl Compounds; Blood Glucose; Diabetes Mellitus, Experimental; Free Radical Scavengers; Glucose Tolerance Test; Hypoglycemic Agents; Inhibitory Concentration 50; Jasminum; Methanol; Nitric Oxide; Phytotherapy; Picrates; Plant Extracts; Plant Leaves; Rats; Rats, Wistar; Reference Standards

The 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity of the fruits of Rubus chingii was studied in vitro. Ethanolic extract, ethyl acetate and n-butanol fractions from dried R. chingii fruits revealed strong DPPH free radical scavenging activity with IC(50) values of 17.9, 3.4 and 4.0 μg/mL, respectively. The ethyl acetate and n-butanol fractions were further purified by a combination of silica gel chromatography, Lobar RP-8 chromatography, and high-pressure liquid chromatography (HPLC). Nine compounds were isolated, where methyl (3-hydroxy-2-oxo-2,3-dihydroindol-3-yl)-acetate (2), vanillic acid (5), kaempferol (7), and tiliroside (9) showed stronger DPPH free radical scavenging activity than that of ascorbic acid (131.8 μM) with IC(50) values of 45.2, 34.9, 78.5, and 13.7 μM, respectively. In addition, rubusine (1) is a new compound discovered in the present study and methyl (3-hydroxy-2-oxo-2,3-dihydroindol-3-yl)-acetate (2), methyl dioxindole-3-acetate (3), and 2-oxo-1,2-dihydroquinoline-4-carboxylic acid (4) were isolated from the fruits for the first time.

Topics: Acetates; Alkaloids; Biphenyl Compounds; Butanols; Chromatography, High Pressure Liquid; Ethanol; Flavonoids; Free Radical Scavengers; Fruit; Oxidation-Reduction; Phenols; Picrates; Plant Extracts; Rubus

Aqueous (ET1) and alcoholic (ET2) extracts from Mactra veneriformis were studied for their antioxidant potentials using various in vitro assays. The ET2 was fractioned into four parts according to the polarity of the extractive medium, viz. ET2-p (petroleum ether), ET2-e (ethyl acetate), ET2-n (n-butanol) and ET2-w (water). Among the four fractions, ET2-w showed the strongest reducing power, and highest 2,2 diphenyl-1-picrylhydrazyl (DPPH) scavenging and metal chelating activities, while the ET2-p possessed higher hydroxyl radical scavenging activities. It was found that ET2-w had large amounts of free amino acids and oligosaccharides, while ET2-p contained a large amount of unsaturated fatty acids. We conclude that amino acids, oligosaccharides and unsaturated fatty acids all contribute to the antioxidant activity of M. veneriformis. In order to elucidate the above, antioxidative components involved in the antioxidant activities were also detected. It was found that amino acids and carbohydrates were the major components of ET2-w. The amino acids were Tau (0.26%), Tyr (0.38%), Met (0.31%), Cys (0.38%), Arg (1.79%) and so on, while carbohydrates were oligosaccharides that were composed of disaccharides and trisaccharides. The unsaturated fatty acids were the major components of ET2-p, which contained a high amount of docosahexaenoic acid (9.03%) and eicosapentaenoic acid (18.49%).

Topics: 1-Butanol; Acetates; Alkanes; Amino Acids; Analysis of Variance; Animals; Antioxidants; Biphenyl Compounds; Bivalvia; Chelating Agents; Complex Mixtures; Docosahexaenoic Acids; Eicosapentaenoic Acid; Fatty Acids, Unsaturated; In Vitro Techniques; Iron; Oligosaccharides; Picrates; Water

The antioxidant activities of the ethanol, petroleum ether, ethyl acetate, n-butanol and water extract fractions from the seeds of papaya were evaluated in this study. The ethyl acetate fraction showed the strongest DPPH and hydroxyl free radical-scavenging activities, and its activities were stronger than those of ascorbic acid and sodium benzoate, respectively. The n-butanol fraction demonstrated the greatest ABTS⁺ radicals scavenging activity. The ethyl acetate fraction and the n-butanol fraction not only showed higher antioxidant activities than the petroleum ether fraction, water fraction and ethanol fraction, but also showed higher superoxide anion and hydrogen peroxide radicals scavenging activities than those of the other extract fractions. The high amount of total phenolics and total flavonoids in the ethyl acetate and n-butanol fractions contributed to their antioxidant activities. The ethyl acetate fraction was subjected to column chromatography, to yield two phenolic compounds, p-hydroxybenzoic acid and vanillic acid, which possessed significant antioxidant activities. Therefore, the seeds of papaya and these compounds might be used as natural antioxidants.

Topics: 1-Butanol; Acetates; Alkanes; Biphenyl Compounds; Carica; Chemistry, Pharmaceutical; Chromatography, Gel; Ethanol; Flavonoids; Free Radical Scavengers; Hydroxyl Radical; Parabens; Picrates; Plant Extracts; Seeds; Solvents; Vanillic Acid; Water

To explore the hepatoprotective and anti-oxidant activities of the methanolic leaf extract of Bridelia micrantha (B. micrantha) on paracetamol induced liver damage in Wistar rats.. Parameters were measured including alanine aminotransaminase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), bilirubin and total protein. The anti-oxidant effects were studied using the 1, 1-Diphenynl-2-Picrylhydrazyl (DPPH) and Ferric Reducing Antioxidant Power (FRAP) assay methods.. B. micrantha extract decreased the level of AST in the rats given PCM from (129.47±0.92I) IU/L to (57.78±1.71) IU/L (P<0.05). This was lower than the value for Silymarin which was (59.92±1.41) IU/L. ALT concentration was reduced from (150.18±2.23) IU/L to (79.10±2.01) IU/L (P<0.05). ALP was reduced from (49.86±0.85) IU/L to (29.64±1.53) IU/L (P<0.05). Total bilirubin was reduced from (2.14±0.10 mg/dL) to (0.18±0.07) mg/dL (P<0.05) while total protein was increased from (4.26±0.30) mg/dL to (6.20±0.19) mg/dL (P<0.05). Concentrations ranging from 10 - 400 μg/mL of B. micrantha were assayed for antioxidant activities. The DPPH assay showed 98% antioxidant activity at concentration of 400 μg/mL. The FRAP values were 0.016, 0.39, 0.455, 0.601 and 1.382 μM at 10, 50, 100, 200 and 400 μg/mL respectively.. Results suggest that B. micrantha has hepatoprotective and anti oxidant potentials. However, further work involving fractionation needs to done to isolate the active compound responsible for the hepatoprotective activity.

Topics: Acetaminophen; Acetates; Alanine Transaminase; Alkaline Phosphatase; Analgesics, Non-Narcotic; Animals; Aspartate Aminotransferases; Bilirubin; Biological Assay; Biphenyl Compounds; Blood Proteins; Chemical and Drug Induced Liver Injury; Dose-Response Relationship, Drug; Euphorbiaceae; Ferric Compounds; Free Radicals; Humans; Male; Phytotherapy; Picrates; Plant Extracts; Plant Leaves; Rats; Rats, Wistar

To determine the acetylcholinesterase inhibitory (AChEI) and antioxidant activity of the ethyl acetate and methanol extracts of 12 traditional medicinal plants used in the treatment of neurological disorders.. AChEI activity was determined spectrophotometrically using the Ellman's colorimetric method. Antioxidant activity was carried out by determining the ability of the extracts to scavenge 2,2-diphenyl-1-picryl hydrazyl (DPPH) and 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radicals. The levels of total phenols, flavonoids and flavonols were determined quantitatively using spectrophotometric methods.. AChEI was observed to be dose-dependent. Lannea schweinfurthii (L. schweinfurthii) (Engl.) Engl. and Scadoxus puniceus (S. puniceus) (L.) Friis & I. Nordal. root extracts showed the lowest IC(50) value of 0.000 3 mg/mL for the ethyl acetate extracts while Zanthoxylum davyi (Z. davyi) (I. Verd.) P.G. Watermann had the lowest IC(50) value of 0.01 mg/mL for the methanol extracts in the AChEI assay. The roots of Piper capense (P. capense) L.f., L. schweinfurthii, Ziziphus mucronata (Z. mucronata) Willd., Z. davyi and Crinum bulbispermum (C. bulbispermum) (Burm.f.) Milne-Redh. & Schweick. showed noteworthy radical scavenging activity and good AChEI activity.. Five plants show good antioxidant and AChEI activity. These findings support the traditional use of the plants for treating neurological disorders especially where a cholinesterase mechanism and reactive oxygen species (ROS) are involved.

Topics: Acetates; Acetylcholinesterase; Africa, Southern; Benzothiazoles; Biphenyl Compounds; Cholinesterase Inhibitors; Colorimetry; Dose-Response Relationship, Drug; Flavonoids; Flavonols; Free Radicals; Fruit; Humans; Inhibitory Concentration 50; Methanol; Phenols; Picrates; Plant Extracts; Plant Roots; Plants, Medicinal; Reactive Oxygen Species; Solvents; Spectrophotometry; Sulfonic Acids

The aim of this study was to investigate the cytoprotective properties of the ethyl acetate fraction of Sargassum muticum (SME) against ultraviolet B (UVB)-induced cell damage in human keratinocytes (HaCaT cells). SME exhibited scavenging activity toward the 1,1-diphenyl-2-picrylhydrazyl radicals and hydrogen peroxide (H(2)O(2)) and UVB-induced intracellular reactive oxygen species (ROS). SME also scavenged the hydroxyl radicals generated by the Fenton reaction (FeSO(4) + H(2)O(2)), which was detected using electron spin resonance spectrometry. In addition, SME decreased the level of lipid peroxidation that was increased by UVB radiation, and restored the level of protein expression and the activities of antioxidant enzymes that were decreased by UVB radiation. Furthermore, SME reduced UVB-induced apoptosis as shown by decreased DNA fragmentation and numbers of apoptotic bodies. These results suggest that SME protects human keratinocytes against UVB-induced oxidative stress by enhancing antioxidant activity in cells, thereby inhibiting apoptosis.

Topics: Acetates; Antioxidants; Apoptosis; Biphenyl Compounds; Catalase; Cell Line; Cell Survival; DNA Fragmentation; Humans; Hydrogen Peroxide; Keratinocytes; Lipid Peroxidation; Oxidative Stress; Picrates; Plant Extracts; Radiation-Protective Agents; Reactive Oxygen Species; Sargassum; Superoxide Dismutase; Ultraviolet Rays

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

A combinative method using high-speed counter-current chromatography (HSCCC) and thin layer chromatography (TLC) as an antioxidant autographic assay was developed to separate antioxidant components from the fruits of Psoralea corylifolia. Under the guidance of TLC bioautography, eight compounds including five flavonoids and three coumarins were successfully separated from the fruits of P. corylifolia by HSCCC with an optimized two-phase solvent system, n-hexane-ethyl acetate-methanol-water (1:1.1:1.3:1, v/v/v/v). The separation produced 5.91mg psoralen, 6.26mg isopsoralen, 3.19mg psoralidin, 0.92mg corylifol A, and 2.43mg bavachinin with corresponding purities of 99.5, 99.8, 99.4, 96.4, and 99.0%, as well as three sub-fractions, in a single run from 250mg ethyl acetate fraction of P. corylifolia extract. Following an additional clean-up step by preparative TLC, 0.4mg 8-prenyldaidzein (purity 91.7%), 4.18mg neobavaisoflavone (purity 97.4%) and 4.36mg isobavachalcone (purity 96.8%) were separated from the three individual sub-fractions. The structures of the isolated compounds were identified by (1)H NMR and (13)C NMR. The results of antioxidant activity estimation by electron spin resonance (ESR) method showed that psoralidin was the most active antioxidant with an IC50 value of 44.7microM. This is the first report on simultaneous separation of eight compounds from P. corylifolia by HSCCC.

Topics: Acetates; Antioxidants; Autoradiography; Biphenyl Compounds; Chromatography, Thin Layer; Coumarins; Countercurrent Distribution; Ethanol; Flavonoids; Fruit; Nuclear Magnetic Resonance, Biomolecular; Picrates; Plant Extracts; Psoralea

On the basis of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay-guided purification, ellagic acid was isolated from the methanol extract of pomegranate fruit peel by liquid-liquid extraction and chromatographic techniques. A reversed-phase high-performance liquid chromatography was described for determination of ellagic acid in pomegranate fruit peel extract. The method involved the use of a TSK-gel ODS-80Tm column with a mixture of 2% aqueous acetic acid and methanol (gradient elution mode: 0-15 min, 40-60% v/v methanol and 15-20 min, 60% v/v methanol) as the mobile phase and detection at 254 nm. The parameters of linearity, repeatability, reproducibility, accuracy, and specificity of the method were evaluated. The recovery of the method was 98.5% and linearity (r(2) > 0.9995) was obtained for ellagic acid. A high degree of specificity as well as repeatability and reproducibility (relative standard deviation values less than 5%) were also achieved. The limits of detection and quantification were 1.00 and 2.50 microg/mL, respectively. The solvent for extraction of ellagic acid from pomegranate fruit peel was examined in order to maximize the ellagic acid content of the extract. A solution of 10% v/v water in methanol was capable of increasing the ellagic acid content in the extract up to 7.66% w/w. The ellagic acid content and antioxidant activity of the ethyl acetate fraction separated from the crude extract using water and ethyl acetate partition was higher than that of the crude extract.

Topics: Acetates; Antioxidants; Biphenyl Compounds; Chemical Fractionation; Chromatography, High Pressure Liquid; Chromatography, Reverse-Phase; Ellagic Acid; Food Analysis; Linear Models; Lythraceae; Methanol; Picrates; Reproducibility of Results; Sensitivity and Specificity

Antioxidant capacities of ethylacetate, butanol, and water fractions of peel, pulp, and seeds of Canarium odontophyllum Miq. (CO) were determined using various in vitro antioxidant models. Ethylacetate fraction of peel (EAFPE) exhibited the highest total phenolic (TPC), total flavonoid content (TFC), and antioxidant activities compared to pulp, seeds, and other solvent fractions. Antioxidant capacities were assayed by total antioxidant capability, 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical activity, ferric reducing antioxidant power (FRAP), and hemoglobin oxidation assay. Total phenolic content of ethylacetate fractions was positively correlated with the antioxidant activity. This is the first report on the antioxidant activities from CO fruit fractions. Thus, EAFPE can be used potentially as a readily accessible source of natural antioxidants and as a possible pharmaceutical supplement.

Topics: Acetates; Analysis of Variance; Antioxidants; Biphenyl Compounds; Burseraceae; Butanols; Chemical Fractionation; Fruit; Hemoglobins; Picrates; Plant Extracts; Seeds; Water

The present study estimates the free radical scavenging activity of the ethyl acetate extract/fractions of Acacia auriculiformis A. Cunn in different assays viz. 1'-1' diphenyl-2'picrylhydrazyl (DPPH), deoxyribose (site specific and non-site specific), relative reducing power, chelating power and lipid peroxidation. The bark powder of the plant was extracted with different solvents by maceration method in the order of increasing and decreasing polarity. The crude ethyl acetate extract was partitioned with ethyl acetate and water (Flow Chart 1 and 2). The scavenging activity of fractions was found to be more as compared to the crude extract. The percent inhibition with water fraction of ethyl acetate extract was observed to be 71.2%, 73.66%, 83.37%, 75.63% and 72.92% in DPPH, chelating power, lipid peroxidation, site specific and non-site specific deoxyribose scavenging assays respectively at maximum concentration tested. l-ascorbic acid and BHT were used as reference compounds for comparing the activity of plant extract/fractions. Studies are in progress to evaluate the effect of extract/fractions in other antioxidant assays and identify the factors responsible for the activity.

Topics: Acacia; Acetates; Biphenyl Compounds; Chelating Agents; Deoxyribose; Dose-Response Relationship, Drug; Free Radical Scavengers; Free Radicals; Hydrazines; Lipid Peroxidation; Oxidation-Reduction; Picrates; Plant Extracts

In the present study, ethyl acetate, butanol and aqueous fractions derived from total methanol extract of Butea monosperma flowers were evaluated for radical scavenging activities using different in vitro models like reducing power assay, scavenging of 2,2 diphenyl-1-picrylhydrazyl (DPPH) radical, nitric oxide radical, superoxide anion radical, hydroxyl radical and inhibition of erythrocyte hemolysis using 2, 2' azo-bis (amidinopropane) dihydrochloride (AAPH). Methanol extract along with its ethyl acetate and butanol fractions showed potent free radical scavenging activity, whereas aqueous fraction was found to be devoid of any radical scavenging properties. The observed activity could be due to the higher phenolic content in the extracts (16.1, 25.29, and 17.74% w/w in methanol extract, ethyl acetate and butanol fractions respectively). HPTLC fingerprint profile of the ethyl acetate and butanol fractions were developed which would serve as reference standard for quality control of the extracts.

Topics: 1-Butanol; Acetates; Biphenyl Compounds; Butea; Erythrocytes; Flowers; Free Radical Scavengers; Free Radicals; Hemolysis; Hydrazines; Hydroxyl Radical; Methanol; Nitric Oxide; Oxidation-Reduction; Picrates; Plant Extracts; Superoxides; Water

The antimutagenic and antioxidant properties of various phenolic fractions obtained from Andean purple corn were examined by the Ames test and the DPPH antiradical assay. An anthocyanin-rich water fraction (WF) and an ethyl acetate fraction (EAF) showed a dose-dependent antimutagenic behavior against the food mutagen Trp-P-1 with IC50 values of 321.7 +/- 21.36 and 95.2 +/- 10.95 microg of chlorogenic acid equiv/plate, respectively, indicating that EAF was a more potent antimutagen. The antioxidant activities for WF and EAF were 1.019 +/- 0.05 and 0.838 +/- 0.11 microg of Trolox equiv/mug of phenolics, respectively. Further fractionation of WF and EAF revealed an ethyl acetate subfraction, EA-IV, with high antimutagen potency that contained a quercetin derivative. The mechanism of antimutagenic action of the WF is predominantly a blocking effect on the S-9 Mix activation system of the mutagen, whereas for the EAF, it is a dual mechanism involving blocking of the S-9 Mix and a scavenging action on Trp-P-1 electrophiles.

Topics: Acetates; Antimutagenic Agents; Antioxidants; Biphenyl Compounds; Free Radicals; Mutagenicity Tests; Phenols; Picrates; Plant Extracts; Water; Zea mays

We evaluated total phenolic content, antioxidant activity, reducing power and antibacterial activity of ethanol, hexane, chloroform, ethyl acetate and aqueous extracts of aerial parts of Rumex japonicus HOUTT. The ethyl acetate extract had the highest amount of phenolic compounds. It also exhibited the highest reducing power and antioxidant activity when assayed by the 1,1-diphenyl-2-picrylhydrazyl (DPPH), beta-carotene bleaching and superoxide radical methods. The ethyl acetate extract possessed the strongest antibacterial activity against Bacillus subtilis, B. cereus and E. coli. GC-MS analysis indicated that ethyl acetate extract contained a variety of phenolic compounds. HPLC analysis showed that pyrogallol was the predominant phenolic compound in this extract. Thus, our study verified that the ethyl acetate extract has strong antioxidant and antibacterial activities which are correlated with its high level of phenolic compounds, particularly pyrogallol and pyrocatechin. This extract of R. japonicus aerial parts can be utilized as an effective and safe source of antioxidants.

Topics: Acetates; alpha-Tocopherol; Anti-Bacterial Agents; Antioxidants; Bacillus cereus; Bacillus subtilis; beta Carotene; Biphenyl Compounds; Butylated Hydroxytoluene; Chromatography, High Pressure Liquid; Gas Chromatography-Mass Spectrometry; Hydrazines; Oxidation-Reduction; Phenols; Picrates; Plant Components, Aerial; Plant Extracts; Rumex; Superoxides

In the course of research on the bioactive constituents of woody plants in the Cyugoku area of Japan, a methanol extract of the leaves of Alnus japonica were found to have strong antioxidative activity. Ethyl acetate soluble and n-buthanol soluble fractions of the methanol extract had a potent antioxidative effect. Both fractions were purified by silica gel column chromatography and HPLC using an ODS column to give four new diarylheptanoids along with known diarylheptanoids and flavonoids. These new compounds were elucidated to be 7-(3,4-dihydroxyphenyl)-5-hydroxy-1-(4-hydroxyphenyl)-3-heptanone-5-O-beta-D-xylopyranoside (1), 1-(3,4-dihydroxyphenyl)-5-hydroxy-7-(4-hydroxyphenyl)-3-heptanone-5-O-beta-D-xylopyranoside (2), 1,7-bis-(3,4-dihydroxyphenyl)-5-hydroxy-3-heptanone-5-O-[2-(2-methylbutenoyl)]-beta-D-xylopyranoside (3) and 1,7-bis-(3,4-dihydroxyphenyl)-5-methoxy-3-heptanone (4) using spectral methods and especially 1H-, 13C-NMR and 2D-NMR measurements. The isolated compounds including their main constituent, oregonin (5), were tested for antioxidative activity. Some of these compounds having two catechol structures showed potent antioxidative activity. Compounds having one catechol structure showed moderate antioxidative activity, but a peracetate of 5 having no catechol structure exhibited no antioxidative activity. Thus the catechol structure of the diarylheptanoids is indispensable for antioxidative activity.

Topics: 1-Butanol; Acetates; Alnus; Antioxidants; Biphenyl Compounds; Chromatography, High Pressure Liquid; Diarylheptanoids; Ethanol; Flavones; Free Radical Scavengers; Magnetic Resonance Spectroscopy; Methanol; Oxidants; Oxidation-Reduction; Picrates; Plant Extracts; Plant Leaves; Solvents; Spectrometry, Mass, Fast Atom Bombardment; Superoxides

Bauhinia monandra Kurz. is used in Brazil for the treatment of diabetes. Since this activity may be correlated with the presence of antioxidant compounds, leaf extracts of B. monandra were evaluated for their radical scavenging capacity (RSC). An ethanolic extract was taken up in aqueous methanol and partitioned with hexane, chloroform, ethyl acetate to yield three organic extracts together with remaining aqueous extract. The RSC was determined spectrophotometrically using 1,1-diphenylpicrylhydrazyl free radical (DPPH). The chloroform and ethyl acetate extracts were the most appropriate as sources of antioxidant compounds as shown by their inhibition concentration (IC50) and inhibition percentage (IP) values. The antioxidant activity of such extracts was attributed to the presence of three compounds of different polarities (flavonoids and steroids). The chloroform and ethyl acetate extracts exhibited an IC50 of approximately 2 mg/g DPPH and IP values in the range of 60-65%. The results indicate that the extracts of B. monandra have a very potent antioxidant activity, compared with the pure catechins used as positive controls and with other plant extracts.

Topics: Acetates; Antioxidants; Bauhinia; Biphenyl Compounds; Chloroform; Chromatography, Thin Layer; Free Radical Scavengers; Hexanes; Hydrazines; Inhibitory Concentration 50; Kinetics; Picrates; Plant Extracts; Plant Leaves; Spectrophotometry

Radical scavenging activities of extracts and constituents in Cornus capitata adventitious root cultures were evaluated by using 1,1-diphenyl-2-pycrylhydrazyl (DPPH) and superoxide anion radicals. Inhibitory activity against peroxidation of linoleic acid was assayed by using the thiobarbituric acid (TBA) method. Ethyl acetate and aqueous fractions were prepared from adventitious roots cultured in Murashige-Skoog liquid medium with 0.1 microM Cu(2+) (0.1CuMS) or 10 microM Cu(2+) (10CuMS). The highest scavenging activities on DPPH and superoxide anion radicals were observed in the ethyl acetate fraction from 0.1CuMS. In the inhibitory activity against linoleic acid oxidation, the ethyl acetate fraction from 10CuMS was highest among the fractions tested. The ethyl acetate fraction of adventitious roots cultured in 0.1CuMS contained mainly galloylglucoses (1,2,3,6-tetragalloylglucose and 1,2,3,4,6-pentagalloylglucose). The ethyl acetate fraction of adventitious roots cultured in 10CuMS contained mainly ellagic acid derivatives [3,3'-di-O-methylellagic acid 4-(5"-acetyl)-alpha-L-arabinofuranoside and stenophyllin H1]. Aqueous fractions prepared from both media contained iridoid glycosides (dihydrocornin and cornin). Tetra- and pentagalloylglucoses showed strong inhibitory activities (61.9 and 85.2%, respectively) against linoleic acid oxidation relative to those of butylated hydroxytoluene (BHT) (91.1%) or alpha-tocopherol (49.5%) at 50 microM concentration. Although both ellagic acid derivatives had weak activities (<50%) on DPPH and superoxide anion radical scavenging, 3,3'-di-O-methylellagic acid 4-(5"-acetyl)-alpha-L-arabinofuranoside was stronger (74.7%) than alpha-tocopherol (49.5%) in inhibiting linoleic acid oxidation at 50 microM concentration. Iridoid glycosides exhibited little activity against DPPH and superoxide anion radicals or against oxidation of linoleic acid.

Topics: Acetates; Antioxidants; Biphenyl Compounds; Cornus; Free Radical Scavengers; Linoleic Acid; Lipid Peroxidation; Picrates; Plant Extracts; Plant Roots; Superoxides; Thiobarbituric Acid Reactive Substances

Since reactive oxygen species (ROS) and hydroxyl radicals (*OH) play an important role in the pathogenesis of many human degenerative diseases, much attention has focused on the development of safe and effective antioxidants. Preliminary experiments have revealed that the methanol (MeOH) extract of the stem of Prunus davidiana exerts inhibitory/scavenging activities on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, total ROS and peroxynitrites (ONOO-). In the present study, the antioxidant activities of this MeOH extract and the organic solvent-soluble fractions, dichloromethane (CH2Cl2), ethyl acetate (EtOAc), and n-butanol (n-BuOH), and the water layer of P. davidiana stem were evaluated for the potential to inhibit *OH and total ROS generation in kidney homogenates using 2',7'-dichlorodihydrofluorescein diacetate (DCHF-DA), and for the potential to scavenge authentic ONOO-. We also evaluated the inhibitory activity of seven flavonoids isolated from P. davidiana stem, kaempferol, kaempferol 7-O-beta-D-glucoside, (+)-catechin, dihydrokaempferol, hesperetin 5-O-beta-D-glucoside, naringenin and its 7-O-beta-D-glucoside, on the total ROS, *OH and ONOO- systems. For the further elucidation of the structure-inhibitory activity relationship of flavonoids on total ROS and *OH generation, we measured the antioxidant activity of sixteen flavonoids available, including three active flavonoids isolated from P. davidiana, on the total ROS and *OH systems. We found that the inhibitory activity on total ROS generation increases in strength with more numerous hydroxyl groups on their structures. Also, the presence of an ortho-hydroxyl group, whether on the A-ring or B-ring, and a 3-hydroxyl group on the C-ring increased the inhibitory activity on both total ROS and *OH generation.

Topics: 1-Butanol; Acetates; Animals; Antioxidants; Ascorbic Acid; Biphenyl Compounds; Catechin; Chromans; Drug Evaluation, Preclinical; Flavonoids; Free Radical Scavengers; Hesperidin; Hydrazines; Hydroxyl Radical; Kaempferols; Kidney; Male; Methanol; Methylene Chloride; Penicillamine; Peroxynitrous Acid; Picrates; Plant Extracts; Plant Stems; Prunus; Rats; Rats, Wistar; Reactive Oxygen Species; Structure-Activity Relationship; Water

Thirty-six different extracts of six herbs and aromatic plants (fennel, common melilot, milfoil, lavandin cv. Super, spike lavender, and tarragon) were evaluated for their radical scavenging activity by the DPPH*, NBT/hypoxanthine superoxide, and *OH/luminol chemiluminescence methods, and for their antioxidant activity by the beta-carotene blenching test. The total phenolic content was also determined by the Folin-Ciocalteu method. The plant material included cultivated plants and their wastes after being distilled for essential oils. Both remarkably high phenolic content and radical scavenging activities were found for the ethyl acetate and dichloromethane fractions among the different plant extracts. In general, the distilled plant material was found to exhibit a higher phenolic content as well as antioxidant and radical scavenging activities than the nondistilled material. Ethyl acetate and dichloromethane extracts, and even some crude extract, of both distilled and nondistilled plants exhibited activities comparable to those of commercial extracts/compounds, thus making it possible to consider some of them as a potential source of antioxidants of natural origin.

Topics: Acetates; Antioxidants; Artemisia; beta Carotene; Biphenyl Compounds; Foeniculum; Free Radical Scavengers; Free Radicals; Hydroxyl Radical; Lavandula; Luminescent Measurements; Luminol; Mediterranean Region; Methylene Chloride; Phenols; Picrates; Plant Extracts

Beans were pearled to evaluate the feasibility of increasing antioxidant activity and phenolic antioxidants. Phenolics were concentrated mostly in the hull fraction at about 56 mg of catechin equivalents per gram of sample. The methanolic extracts of the pearled bean samples were screened for antioxidant potential using the beta-carotene-linoleate and the 1,1-diphenyl-2-picrylhydrazyl (DPPH) in vitro model systems. The pearled material, also referred to as milled samples, exhibited antioxidant activity that correlated with phenolic content and inhibited DPPH significantly in a dose-dependent manner. Phenolics and antioxidant activities were also examined in chromatographic fractions of methanolic extracts of manually obtained hulls that represented a model used previously to ascertain antimutagenic activity. Fractions extracted with ethyl acetate/acetone and acetone displayed antioxidant activity, which implies potent free radical scavenging activity with antimutagenic activity.

Topics: Acetates; Acetone; Antimutagenic Agents; Antioxidants; beta Carotene; Biphenyl Compounds; Free Radical Scavengers; Kinetics; Linoleic Acid; Methanol; Phaseolus; Phenols; Picrates; Plant Extracts; Seeds

Residues of the oil-extracting process of oilseeds contain bioactive substances such as phenolic compounds, which could be used as natural antioxidants for the protection of fats and oils against oxidative deterioration. Thus, the extraction of such constituents from residual material can be considered to contribute to the added value of these residues, which could justify their isolation. In the present work the fat-free residues of eight different oilseeds whose oils are usable for nutritional applications, and also as renewable resources, were extracted with 70% methanol, 70% acetone, water, and ethyl acetate/water, respectively. The resulting extracts were investigated regarding their content of total phenolic compounds by the Folin-Ciocalteau assay, sinapine, flavanoids, and the UV-absorption spectra. Further, the antioxidant activity of the extracts was characterized by the DPPH method, the beta-carotene-linoleic acid assay, and ESR investigations. The fat-free residues of the different oilseeds contained considerable amounts of extractable substances. The yields decreased with decreasing polarity of the solvent in the order water, 70% methanol, 70% acetone, and ethyl acetate. The ratio of total phenolic compounds to the extractable compounds ranged from 3 to 19%. There was no significant correlation between the amount of total extractable compounds and the total phenolic compounds (p < 0.001). All extracts showed remarkable antioxidant activities determined with the different methods. The effects depended strongly on the solvent used for the extraction as well as on the extracted residue. A correlation between the methods used for the characterization of the antioxidant activity and the composition of the residues could not be shown.

Topics: Acetates; Acetone; Antioxidants; beta Carotene; Biphenyl Compounds; Choline; Electron Spin Resonance Spectroscopy; Flavonoids; Free Radical Scavengers; Methanol; Phenols; Picrates; Plant Oils; Seeds; Water