Antioxidant, antimicrobial, and anticancer activity of 3 Umbilicaria species

Abstract: The aim of this study is to investigate in vitro antioxidant, antimicrobial, and anticancer activity of the acetone extracts of the lichens Umbilicaria crustulosa, U. cylindrica, and U. polyphylla. Antioxidant activity was evaluated by 5 separate methods: free radical scavenging, superoxide anion radical scavenging, reducing power, determination of total phenolic compounds, and determination of total flavonoid content. Of the lichens tested, U. polyphylla had largest free radical scavenging activity (72.79% inhibition at a concentration of 1 mg/mL), which was similar as standard antioxidants in the same concentration. Moreover, the tested extracts had effective reducing power and superoxide anion radical scavenging. Total content of phenol and flavonoid in extracts was determined as pyrocatechol equivalent, and as rutin equivalent, respectively. The strong relationships between total phenolic and flavonoid contents and the antioxidant effect of tested extracts were observed. The antimicrobial activity was estimated by determination of the minimal inhibitory concentration by the broth microdilution method. The most active was extract of U. polyphylla with minimum inhibitory concentration values ranging from 1.56 to 12.5 mg/mL. Anticancer activity was tested against FemX (human melanoma) and LS174 (human colon carcinoma) cell lines using MTT method. All extracts were found to be strong anticancer activity toward both cell lines with IC₅₀ values ranging from 28.45 to 97.82 μg/mL. The present study shows that tested lichen extracts demonstrated a strong antioxidant, antimicrobial, and anticancer effects. That suggests that lichens may be used as possible natural antioxidant, antimicrobial, and anticancer agents.     

Antioxidant Activities In Vitro of Ethanol Extract and Fractions from Mushroom,Lenzites Betulina

The antioxidant potential of different fractions of Lenzites betulina was investigated. Fraction 2 showed the highest antioxidant activity in the 2, 2‐diphenyl‐1‐picrylhydrazyl (DPPH) and hydroxyl (OH) radical‐scavenging assays. Three natural p‐terphenyls were identified in this fraction. The structure of a new natural compound was established as 2‐phenyl‐[1H‐2‐benzopyran][4,3‐e][p]benzoquinone in addition to two other compounds, 2,5‐diphenyl‐3,6‐dimethoxy‐p‐benzoquinone and 2‐phenyl‐3‐methoxy‐[1H‐2‐benzopyran][4,3‐e][p]benzoquinone that have been previously reported in L. betulina. The antioxidant activities of these p‐terphenyls were evaluated by DPPH and OH radical‐scavenging assays. 2‐phenyl‐3‐methoxy‐[1H‐2‐benzopyran][4,3‐e][p]benzoquinone was the most active in the OH radical‐scavenging test (IC ₅₀ = 37.69 μg/mL) and showed little inhibition in the DPPH assay. The chemical components of L. betulina therefore have some antioxidant capacities, and this species could be used as a potential source of new natural antioxidants.     

不同极性柳叶蜡梅叶萃取物总酚含量及其抗氧化、抑菌能力研究

采用甲醇回流提取、梯度萃取得到正己烷萃取物、二氯甲烷萃取物、乙酸乙酯萃取物、正丁醇萃取物和水萃取物5个不同极性部分,以多酚含量、还原能力、对1,1-二苯基-2-三硝基苯肼(DPPH)自由基的清除能力及抑制微生物生长为评价指标,研究其体外抗氧化和抑菌活性能力。结果表明:柳叶腊梅叶多酚量较为丰富,乙酸乙酯萃取物含量最多,为(309.5±4.81)mg/g。柳叶腊梅叶萃取物具有较强的抗氧化活性,其还原能力和对DPPH自由基清除能力随其浓度的增加而增强。甲醇、正己烷、二氯甲烷、乙酸乙酯、正丁醇和水萃取物对DPPH自由基清除能力的半数抑制浓度(IC50)值分别为1.24,1.11,0.61,0.48,0.98,1.36 mg/mL。柳叶腊梅叶萃取物对大肠杆菌、金黄葡萄球菌、枯草芽孢杆菌和绿脓杆菌均有抑制作用,二氯甲烷萃取物的抑菌活性最显著,其对4种受试菌的最低抑制浓度为2 mg/mL。故柳叶腊梅叶萃取物具有较好的抗氧化和抑菌特性,为进一步分离提取活性化合物提供依据。     

苦丁茶提取物的分离纯化、鉴定及不同萃取部位活性分析

以苦丁茶为原料，对其乙醇提取物和不同溶剂萃取部位的抗氧化活性、抗补体活性、抑制癌细胞活性进行探究，并对活性最佳的萃取部位进一步分离纯化，分析鉴定出单体化合物。抗氧化活性以总还原力、清除1,1-二苯基-2-三硝基苯肼自由基和超氧阴离子自由基的能力为指标，通过溶血实验评价抗补体活性，并采用MTT法测定对人乳腺癌细胞MCF-7的存活率，结合多种分离技术获得不同单体化合物，通过对质谱、核磁共振波谱的数据分析鉴定其化学结构。结果表明，苦丁茶乙醇提取物及各部位均具有良好的抗氧化效果，其中乙酸乙酯部位不仅在实验质量浓度范围内有较强的抗氧化活性（0.8 mg/mL均达到90%以上），还具有良好的抗补体活性（79.67±0.99）%，能很好抑制人乳腺癌MCF-7细胞增殖活性（54.8±0.26）%，对乙酸乙酯部位进行分离纯化，总共分离得到11 个单体化合物。由此可见，苦丁茶乙醇提取物和各部位均具有抗氧化、抗补体活性和人乳腺癌MCF-7细胞抑制活性，乙酸乙酯作为萃取剂时活性最佳。分离的单体化合物鉴定后多为酚类、黄酮类物质。     

Antioxidant, antimicrobial and anticancer activities of three Parmelia species

BACKGROUND: Lichens are symbiotic organisms consisting of algae and fungi. They are used for human and animal nutrition and in the production of colours, perfumes and alcohol. Lichens have also been used in traditional medicine to treat diseases such as jaundice, pulmonary, stomach and cranial diseases. In this study the acetone extracts of three lichens, Parmelia caperata, Parmelia sulcata and Parmelia saxatilis, were tested for their antioxidant, antimicrobial and anticancer potential. RESULTS: Of the lichens tested, P. saxatilis had the highest free radical‐scavenging activity (55.3% inhibition). Moreover, all tested extracts showed effective reducing power and superoxide anion radical scavenging. Strong relationships between total phenolic and flavonoid contents and antioxidant effects of the tested extracts were observed. The extract of P. sulcata was most active in terms of antimicrobial ability, with minimum inhibitory concentration values ranging from 0.78 to 12.5 mg L^?1. All extracts were found to have strong anticancer activity, with IC₅₀ values ranging from 9.55 to 22.95 µg mL^?1. CONCLUSION: The present study showed that the tested lichen extracts exhibited strong antioxidant, antimicrobial and anticancer effects. This suggests that lichens may be used as possible natural antioxidant, antimicrobial and anticancer agents. Copyright © 2012 Society of Chemical Industry     

Antioxidant and DNA protection activities of a glycoprotein isolated from a seaweed,Saccharina japonica

A glycoprotein with a molecular weight of ～10 kDa was purified from Saccharina japonica, and its antioxidant activity was evaluated using five different antioxidant assays. The highest antioxidant activities of the purified glycoprotein determined by DPPH, ABTS, SODA, SORS and FRAP assays were 85%, 95%, 94%, 72% and 97%, respectively. About 37% nitric oxide scavenging and 85% xanthine oxidase inhibition activities were also observed at a concentration of 5 mg mL^?1. The glycoprotein exhibited optimal activity levels of free radical scavenging at pH 7–9 and 40 °C. Antioxidant activity decreased when treated with ethylenediaminetetraacetic acid and metal ions. Moreover, its scavenging activity decreased when the glycoprotein was treated with deactivation agents such as pronase E or sodium periodate (NaIO₄), and it showed protective activity against DNA cleavage induced under oxidative conditions.     

Antioxidant activity of compounds isolated from the pyroligneous acid, Rhizophora apiculata

The polyphenols (CPAE II) was isolated from the dichloromethane extract of the pyroligneous acid, Rhizophora apiculata by simultaneous acid base and solvent extraction method. Its qualitative and quantitative composition was studied by gas chromatography mass spectroscopy (GC/MS) and out of 57 peaks, 52 compounds were identified, representing 95.47% of the total polyphenols. The CPAE II was then fractionated to four fractions (F1–F4) by means of thin layer chromatography and silica gel column chromatography with dichloromethane, dichloromethane/chloroform/ethyl acetate mixture (8:1:1; 4:3:3, v/v/v), and ethyl acetate, respectively. The antioxidant properties of the CPAE II and the fractions were evaluated. Among the four fractions, fraction 1 (F1) was the most potent in DPPH radical scavenging activity and molybdenum (VI) reducing power. It was subjected to further purification by means of silica gel column chromatography with hexane, hexane/diethyl ether mixture (9:1, 6:1, 3:1, v/v), and diethyl ether, respectively. 2,6-Dimethoxyphenol (syringol) and dihydroxybenzenes (catechol and 3-methoxycatechol) were isolated and identified by GC/MS, ¹H NMR, ¹³C NMR spectral analyses, and confirmed by GC co-injection with authentic standards. Syringol, catechol and 3-methoxycatechol constitute 39.08, 4.21 and 1.10% of F1, respectively. Their antioxidant activities were evaluated by DPPH radical scavenging activity, ABTS radical cation scavenging activity, phosphomolybdenum and ferric reducing antioxidant power (FRAP). The trend in antioxidant capacity was similar in all the four assays, with dihydroxybenzenes > 2,6-dimethoxyphenols, although discrepancies in the ranking within the dihydroxybenzenes were present. These three compounds which showed significant antioxidant activities were isolated for the first time from the pyroligneous acid, R. apiculata.     

Cladonia lichens and their major metabolites as possible natural antioxidant,antimicrobial and anticancer agents

The aim of this study is to investigate in vitro antioxidant, antimicrobial, and anticancer activities of the acetone extracts of the lichens Cladonia furcata, Cladonia pyxidata and Cladonia rangiferina and their atranorin and fumarprotocetraric acid constituents. Antioxidant activity was evaluated by free radical scavenging, superoxide anion radical scavenging, reducing power, and determination of total phenolic and flavonoid compounds. As a result of the study atranorin had largest free radical scavenging activity with IC₅₀ values 131.48 μg/mL. Moreover, the tested samples had effective reducing power and superoxide anion radical scavenging. Total content of phenol and flavonoid in extracts was determined as pyrocatechol equivalent, and as rutin equivalent, respectively. The strong relationships between total phenolic and flavonoid contents and the antioxidant effect of tested extracts were observed. The antimicrobial activity was estimated by determination of the minimal inhibitory concentration by the broth microdilution method. The most active was fumarprotocetraric acid with minimum inhibitory concentration values ranging from 0.031 to 0.125 mg/mL. Anticancer activity was tested against FemX (human melanoma) and LS174 (human colon carcinoma) cell lines using MTT method. All samples were found to be strong anticancer activity toward both cell lines with IC₅₀ values ranging from 10.97 to 41.23 μg/mL.     

Identification of antioxidant compounds of Mucuna sempervirens by high-speed counter-current chromatographic separation-DPPH radical scavenging detection and their oestrogenic activity

High-speed counter-current chromatography (HSCCC) was used to separate an ethanolic extract of leaves of Mucuna sempervirens into fractions which were then detected their antioxidant activity using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. The fractions were grouped into seven larger fractions (components) based on their antioxidant activity. The seven components were isolated by preparative HPLC to yield 12 flavonoids and two phenolic acids identified by electrospray ionisation mass spectrometry and nuclear magnetic resonance analyses. The oestrogenic activity of the 12 isolated flavonoids was evaluated by the luciferase assay based on the MVLN cell line. The results indicate that leaves of M. sempervirens are a flavonoid-rich resource that may supply antioxidant and oestrogenic compounds to the human body. The HSCCC separation-DPPH radical scavenging detection could be widely applied for rapid screening and isolation of antioxidants from complex plant extracts.     

Antioxidant and antimicrobial activities of Polygonum cognatum Meissn extracts

The antioxidant activities, reducing powers, 2,2‐diphenyl‐l‐picrylhydrazyl (DPPH) radical‐scavenging activities, total phenolic compound contents and antimicrobial activities of ether, ethanol and hot water extracts of Polygonum cognatum Meissn were studied in vitro. The highest antioxidant activity was found in the water extract. However, there were no statistically significant differences among 15 µg ml^?1 extract‐containing samples in linoleic acid emulsion (0.02 M , pH 7.0) during 120 h of incubation (P > 0.05). The reducing power of the water extract was the highest, but its reducing power was markedly lower than that of ascorbic acid. The highest DPPH radical‐scavenging activity was found in the water extract, with 50% DPPH radical scavenging at a concentration of 100 µg ml^?1 dried water extract, while at the same concentration of dried ethanol extract the value was 12%. Surprisingly, no DPPH radical‐scavenging activity was observed in the ether extract. The concentrations of phenolic compounds found were 0.48, 0.50 and 0.01 µg ml^?1 gallic acid equivalent in 10 µg ml^?1 water, ethanol and ether extracts respectively. The ether and ethanol extracts showed antimicrobial activity against Staphylococcus aureus and Bacillus subtilis. The water extract did not show antimicrobial activity against the studied micro‐organisms. © 2002 Society of Chemical Industry     

何首乌中二苯乙烯苷的体外抗氧化研究

从传统抗衰老中药何首乌中提取、分离、纯化得到不同组分:乙醇粗提物、大孔吸附树脂层析样品及二苯乙烯苷晶体,并运用HPLC-MS、NMR、X-衍射对二苯乙烯苷晶体进行了分析鉴定。对照白藜芦醇分别对三组分的抗氧化清除自由基性能进行了测定。包括还原能力、亚油酸体系抗氧化性能以及清除DPPH自由基、清除羟基自由基、清除超氧阴离子能力。结果表明:不同纯度的二苯乙烯苷均具有还原性,抗氧化及清除自由基性能,其中以二苯乙烯苷纯品最强。其半抑制率分别为:DPPHIC508.06μg/ml;羟基自由基IC502.75μg/ml;超氧阴离子IC500.57μg/ml;以上各指标均略高于白藜芦醇。     

Identification of radical scavenging peptides (<3 kDa) from Burgos-type cheese

The aim of the present study was to identify low molecular weight peptides with radical scavenging activity from cheese made using different types of rennet: batch 1, animal rennet (95% chymosin and 5% bovine pepsin), batch 2, rennet of plant origin (Cynara cardunculus) and batch 3, microbial rennet (Mucor miehei). After preparation of the peptide extracts (<3 kDa), antioxidant activity was assayed by their DPPH radical scavenging and metal chelating a6ctivity. All of the batches showed antioxidant activity, which could be dependent on the peptides which are present in extracts: Batch 2 and 3 showed the highest values for DPPH inhibition and chelating effect. Fourteen fractions out of the total peptide fractions collected after RP-HPLC analysis showed radical scavenging activity using the DPPH inhibition method. Free amino acids and peptides were identified from these fractions. One of the peptides, derived from α_s1-casein, was a potential new antioxidant peptide. These antioxidant peptides were present in a lower content in extracts obtained from animal rennet cheese in comparison with the other extracts.     

Lipophilic phenols partially explain differences in the antioxidant activity of subfractions from methanol extract of camellia oil

The crude methanol extract (ME) from camellia oil was fractionated by serial benzene, ether, EtOAc and n-BuOH, divided into five subfractions designated as SF1–5. The obtained fractions as well as ME were evaluated for their antioxidant activities using DPPH assay, and their phenolic constituents were isolated using silica gel column chromatography and analysed by TLC, RP-HPLC–PDA, UPLC-ESI–MS and GC–MS. Results showed that the content of total phenolics (TPs) of ME, determined by the Folin–Ciocalteu method, was estimated to be 79.5?±?10.2?mg of p-hydroxybenzoic acid equivalent/kg oil, while that of the total flavonoids was 8.98?±?0.54?mg of rutin equivalent/kg oil. ME exhibited pronounced radical-scavenging activity against the stable DPPH radical (with an antioxidant capacity IC₅₀ value of 52.37?μg/mL), and three subfractions (SF1–3) separated from ME contributed the most significant activity with IC₅₀ of 28.41, 17.42 and 43.17?μg/mL, respectively. Consistent with the different capacity of scavenging DPPH radicals, the subfractions from ME showed different composition and contents of phenolic compounds as detected by TLC, RP-HPLC–PDA, UPLC-ESI–MS and GC–MS. Kaempferol, as well as a number of volatile phenolic compounds, was first identified from camellia oil. Results showed that the presence of phenolic compounds and flavonoids in each extracts may be responsible for its individual antioxidant activity, and the lipophilic phenolic fractions would likely account for a great part of the antioxidant properties of the oil.     

Determination of Chemical Composition,Total Phenolic,Antimicrobial, and Antioxidant Activities of Echinophora tenuifolia Essential Oil

In this study, the antioxidant and antimicrobial activities, total phenolic content, and essential oil composition of Echinophora tenuifolia L. subsp. sibthorpiana were investigated. The antioxidant activity of investigated essential oil was assessed by ABTS and DPPH assays. DPPH radical scavenging activity expressed by IC₅₀ was 2.84 g/L, whereas the TEAC value determined by ABTS assay was 0.032 g TEAC/kg plant. Total phenol content of essential oil determined by Folin-Ciocalteu method was calculated as 1.32 g GAE/kg plant. The essential oil extracted by hydrodistillation (Clevenger apparatus) was investigated by GC-MS technique and 78 compounds were identified. The main components of essential oils were found to be δ-3-carene (17.93%), p-cymene (8.99%), methyleugenol (16.41%), and α-phellandrene (9.33%). The antimicrobial activity of investigated essential oil was tested using a broth dilution method against 13 bacterial and 2 fungal microorganisms. The lowest minimum inhibitory concentration of essential oil against Bacillus cereus was 62.5 μg/mL while the antifungal activity was greater than 1000 μg/mL for both Candida albicans and Saccharomyces cereviciae. Investigated essential oil has a certain level of antioxidant and antimicrobial effects, which may be attributed to their chemical compounds. The antimicrobial efficiency of essential oil, especially against Bacillus cereus and Staphylocoocus spp., offers its effectiveness to treatment of wound or disease caused by Gram positive bacteria.     

Compositions, antioxidant and antimicrobial activities of Helichrysum (Asteraceae) species collected from Turkey

The methanolic extracts of 16 Helichrysum species were investigated for their in vitro antioxidant, radical scavenging and antimicrobial activities. All the extracts showed strong antioxidant and radical scavenging activity. The highest total antioxidant capacity as ascorbic acid equivalents (AAE) of 194.64 mg/g dry extract was obtained for Helichrysum noeanum in the phosphomolybdenum assay. The highest IC₅₀ value (7.95 μg/ml) was observed for the extract of Helichrysum stoechas subsp. barellieri in the DPPH assay. The total phenolic contents of the extracts ranged from 66.74 to 160.63 mg gallic acid equivalents (GAE)/g dry extract. The major component present in the extracts was identified as chlorogenic acid followed by apigenin-7-glucoside and apigenin by HPLC analysis. All the extracts showed significant antimicrobial activity against microorganisms containing 13 bacteria and two yeasts in the agar diffusion method.     

Bioactive compounds and antioxidant potential in fresh and dried Jaffa® sweeties,a new kind of citrus fruit

Bioactive compounds in the edible parts of fresh and dried Jaffa^® sweeties, a new kind of citrus fruit, were analysed and their antioxidant capacities were assessed. Antioxidant‐rich fractions were extracted from fresh and dried sweeties with 1.2 M HCl in methanol/water (1:1 v/v), and the antioxidant activities of these extracts were evaluated. Using the β‐carotene/linoleate model system, the extracts from equivalent quantities of fresh and dried sweeties showed 89 and 87% antioxidant activity respectively. Similarly, using the DPPH radical‐scavenging method, the extracts showed 87 and 85% antioxidant activity respectively. The best correlations were between caffeic acid content and β‐carotene and DPPH antioxidant activity values (r = 0.9849 and 0.9798 respectively, p = 0.005). Both fresh and dried sweeties are bioactive natural products; when fresh fruits are not available, properly dried sweeties could be used as a substitute. Copyright © 2004 Society of Chemical Industry     

Isolation and identification of an antioxidant flavonoid compound from citrus-processing by-product

BACKGROUND: Large amounts of citrus by‐products are released from juice‐processing plants every year. Most bioactive compounds are found in the peel and inner white pulp. Flavonoids are a widely distributed group of bioactive compounds. The methanolic extract of citrus peel powder has been shown to possess strong antioxidant activity. Therefore the aim of this study was to isolate the major antioxidant flavonoid compound from Citrus unshiu (satsuma) peel as citrus by‐product and evaluate its antioxidant activity. RESULTS: The major flavonoid isolated from C. unshiu peel was identified as quercetagetin. The structure of the compound was determined by tandem mass spectrometry and ultraviolet spectroscopy. Its antioxidant activity was assessed by assays of 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) radical, hydroxyl radical and intracellular reactive oxygen species (ROS) scavenging and DNA damage inhibition. Quercetagetin showed strong DPPH radical‐scavenging activity (IC₅₀7.89 µmol L^?1) but much lower hydroxyl radical‐scavenging activity (IC₅₀203.82 µmol L^?1). Furthermore, it significantly reduced ROS in Vero cells and showed a strong protective effect against hydrogen peroxide‐induced DNA damage. CONCLUSION: The results of this study suggest that quercetagetin could be used in the functional food, cosmetic and pharmaceutical industries. Copyright © 2011 Society of Chemical Industry     

Antioxidant phenolic compounds from Smilax sebeana Miq.

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

Evaluation of Synurus deltoides for antioxidant,antimicrobial, and anti-proliferative activities using In vitro assays

The antioxidant, antimicrobial, and anti-proliferative activities of water and ethanol extracts from leaves of Synurus deltoides (SD) were investigated. Extracts showed strong reductive powers, nitrite-scavenging activities, DPPH, ABTS, hydroxyl, and superoxide radical scavenging activities, and DNA damage prevention activities. SD extracts also showed strong cellular antioxidant activities in a dose-dependent manner with IC₅₀ values of 0.93 and 0.65 mg/mL for water and ethanol extracts, respectively. Antimicrobial activities were evaluated against 4 microorganisms. SD extracts showed high inhibitory activities against Pseudomonas aeruginosa and Staphylococcus aureus. The anti-proliferative activity was dose-dependent against human colon adenocarcinoma (HT-29) and mouse rectum carcinoma cells (CMT-93). In addition, HPLC analyses of SD leaf extracts revealed the presence of different phenolic compounds. SD can be considered as a source of functional foods and pharmaceuticals.