Electron spin resonance spectroscopy studies on the free radical scavenging activity of wine anthocyanins and pyranoanthocyanins

Anthocyanins are a group of natural occurring pigments responsible for the red-blue color of grapes and many fruits and vegetables. Anthocyanins and derived pigments are of double interest, one technological, as they can be used as natural colorants, and another one due to their implication on human health through their antioxidant activity. Although there are numerous studies regarding the antioxidant activity of grape extracts as well as red wine, the free radical scavenging activity of purified anthocyanins and pyranoanthocyanins is largely unknown. In the present study, the hydroxyl and superoxide anion scavenging activities of anthocyanins and their pyruvic acid adducts were systematically investigated by electron spin resonance spectroscopy and spin trapping. The 3-glucosides of delphinidin, cyanidin, petunidin, pelargonidin and malvidin, and the pyruvic adduct of the 3-glucoside of delphinidin exhibited a potent superoxide anion radical scavenging and, to a lesser extent hydroxyl anion radical scavenging activity. The pyranoanthocyanins of cyanidin, petunidin, malvidin and pelargonidin showed a high capacity to scavenge superoxide anion radicals but did not scavenge hydroxyl radicals. Current data indicate that formation of anthocyanin adducts with pyruvic acid, which may occur during wine ageing or fruit juice processing, decreases the hydroxyl and superoxide anion scavenging and thus could decrease the antioxidant potential of these compounds.     

Changes in free-radical scavenging ability of kombucha tea during fermentation

Kombucha tea is a fermented tea beverage produced by fermenting sugared black tea with tea fungus (kombucha). Free-radical scavenging abilities of kombucha tea prepared from green tea (GTK), black tea (BTK) and tea waste material (TWK) along with pH, phenolic compounds and reducing power were investigated during fermentation period. Phenolic compounds, scavenging activity on DPPH radical, superoxide radical (xanthine–xanthine oxidase system) and inhibitory activity against hydroxyl radical mediated linoleic acid oxidation (ammonium thiocyanate assay) were increased during fermentation period, whereas pH, reducing power, hydroxyl radical scavenging ability (ascorbic acid–iron EDTA) and anti-lipid peroxidation ability (thiobarbituric assay) were decreased. From the present study, it is obvious that there might be some chances of structural modification of components in tea due to enzymes liberated by bacteria and yeast during kombucha fermentation which results in better scavenging performance on nitrogen and superoxide radicals, and poor scavenging performance on hydroxyl radicals.     

荷叶功能成分的提取及其对自由基清除作用的研究

研究了荷叶中的功能成分黄酮类化合物和生物碱的水提取工艺条件。结果表明 :以水为溶剂来提取荷叶中的黄酮类化合物及生物碱时 ,较佳的工艺条件为 ,m (水 )∶m(荷叶 ) =30∶1,于 80～ 90℃提取 1.5h ,提取液呈酸性。同时采用电子自旋共振捕集技术 (ESR) ,研究了以荷叶在水溶剂中的提取物 (LLE)对羟基自由基 (·OH)和超氧阴离子自由基 (O-2 ·)的清除效果。结果表明 :LLE对·OH和O- 2 ·有很强的清除能力 ,2 6.94 μg/mL的LLE对次黄嘌呤 黄嘌呤氧化酶体系产生O-2 ·的清除率达 65 .60 % ,LLE浓度大于 8.98mg/mL时 ,可以全部清除由Fenton反应体系产生的·OH。     

Antioxidant activities of red pepper (Capsicum annuum) pericarp and seed extracts

In this study, we examined the antioxidant activities of red pepper (Capsicum annuum, L.) pericarp and red pepper seed extracts. The extracts were evaluated by various antioxidant assays, including 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) radical scavenging, superoxide anion radical scavenging, hydroxyl radical scavenging, [2,2′‐azino‐bis(3‐ethylbenzthiazoline‐6‐sulphonic acid)] (ABTS) radical scavenging, ferrous chelating activity, superoxide dismutase (SOD) activity and reducing power, along with the determination of total phenolic and flavonoid contents. All the extracts showed strong antioxidant activity by the testing methods. The red pepper pericarp extract exhibited strong ferrous chelating activity and high scavenging activity against free radicals, including both the hydroxyl and DPPH radicals, but it exhibited weaker scavenging activity for the superoxide anion radical and for SOD. In contrast, the red pepper seed extract exhibited strong SOD activity and high scavenging activity against the superoxide anion radical, but showed weaker ferrous chelating activity, hydroxyl radical scavenging, and DPPH radical scavenging. We observed that the reducing power level and ABTS radical scavenging activity of the red pepper seed were higher than those of the red pepper pericarp at the highest tested concentration. Most of the test results for the red pepper seed and red pepper pericarp extracts increased markedly with increasing concentration; however, the metal chelating, SOD and ABTS radical scavenging activities did not increase with the concentration. Highest total phenolic and flavonoid contents were obtained from the red pepper pericarp extracts. Overall, the red pepper seed and red pepper pericarp extracts were highly effective for the antioxidant properties assayed, with the exceptions of ferrous chelating activity, hydroxyl radical scavenging and SOD activity.     

Identification of phenolic compound in manuka honey as specific superoxide anion radical scavenger using electron spin resonance (ESR) and liquid chromatography with coulometric array detection

Apitherapy has become the focus of attention as a form of folk and preventive medicine for treating certain conditions and diseases as well as promoting overall health and well‐being. In apitherapy, honey is the therapeutic agent used for dressing surgical wounds, burns or skin ulcers, as well as for dyspepsia, peptic ulcer, etc., because of its antioxidant activity. Therefore, it is important to determine the antioxidants in honey by analytical techniques. In the present study, the antioxidant activities of honeys from different floral sources were investigated by electron spin resonance (1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) and H₂O₂/NaOH/DMSO scavenging systems), liquid chromatography with coulometric array detection (LC‐ED), and liquid chromatography with electrospray mass spectrometry (LC‐MS). The antioxidant activities of some unifloral honeys (acacia, Chinese milk vetch, buckwheat and manuka) were evaluated using the radical scavenging systems. It was shown that DPPH radical scavenging activity was significantly different among the honeys, with buckwheat and manuka honeys having significantly higher scavenging activity than acacia honey. In addition, only manuka honey had specific scavenging activity for superoxide anion radicals. The compound responsible for this activity in manuka honey was identified by LC‐ED and LC‐MS. Careful examination of the LC‐ED chromatographic patterns of manuka and other honey samples revealed a distinct peak in the chromatogram of manuka honey to be methyl syringate (MSYR). The radical scavenging activity of MSYR was specific for superoxide anion radicals, similar to the case of manuka honey. Copyright © 2005 Society of Chemical Industry     

Antioxidant properties of herbal extracts selected from screening for potent scavenging activity against superoxide anions

BACKGROUND: In a previous study with the electron spin resonance (ESR) spin‐trapping method on large‐scale screening with ethanol extracts of approximately 1000 kinds of herbs, four herbal extracts with prominently potent ability to scavenge superoxide anions were chosen, namely the extracts from Punica granatum (peel), Syzygium aromaticum (bud), Mangifera indica (kernel) and Phyllanthus emblica (fruit)). In the present study, these extracts were further examined to determine if they also scavenge hydroxyl radicals, by applying the ESR spin‐trapping method, and if they have heat resistance as a desirable characteristic feature. RESULTS: Experiments with the Fenton reaction and UV irradiation of hydrogen peroxide, both of which generate hydroxyl radicals, showed that all four extracts had potent ability to directly scavenge hydroxyl radicals. Each extract exerted more potent hydroxyl radical‐scavenging activity than mannitol and salicylic acid. Furthermore, the scavenging activities against superoxide anions and hydroxyl radicals of the extracts of P. granatum (peel), M. indica (kernel) and P. emblica (fruit) proved to be heat‐resistant. CONCLUSION: The four herbal extracts chosen from extensive screening possess desirable antioxidant properties. In particular, three of them are expected to be suitable for food processing in which thermal devices are used, because of their heat resistance. Copyright © 2008 Society of Chemical Industry     

杜仲素抗氧化活性研究

以VC 为阳性对照,对杜仲素和高纯绿原酸清除羟自由基(·OH)、超氧阴离子自由基(O2·)及1,1- 二苯基-2- 苦苯肼自由基(DPPH 自由基)活性进行研究,并通过计算清除率比较清除各种自由基活性的效果。结果表明：杜仲素在不同自由基体系中抗氧化活性不同,清除自由基能力为O2·＞ DPPH 自由基>·OH,对3 种自由基的最大清除率依次为95.85%、75.31% 和31.10%。因此,杜仲素具有较强的抗氧化作用。     

Comparative study on composition and antioxidant properties of mint and black tea extract

The antioxidant properties of plants could be correlated with oxidative stress defence in different human diseases. The present study was undertaken to evaluate and compare the antioxidant potential and the phytochemical composition in the aqueous extracts of mint leaves, black tea and black tea enriched with mint extract. All the three preparations exhibited free radical‐scavenging potential for nitric oxide (NO) radical, superoxide anion radical and hydroxyl radical, and the values were lesser than those of the antioxidants which acted as standards. In comparison, the mint extract exhibited higher free radical and NO scavenging effect. Hydroxyl radical and superoxide scavenging effects were more pronounced in tea with the mint extract, while the reducing power was exhibited more significantly by the black tea extract. The phytochemical compounds were identified and the total phenols and flavonoids were quantified and compared between these extracts.     

不同花色种类白茶的抗氧化活性及其主要品质化学成分分析

目的：分析不同花色种类白茶（白毫银针、白牡丹和寿眉）茶汤的抗氧化活性、茶汤中的主要品质化学成分以及二者的相关性。方法：采用铁还原抗氧化能力、清除1,1-二苯基-2-三硝基苯肼自由基活性、清除2,2’-联氮基-双-（3-乙基苯并噻唑啉-6-磺酸）二铵盐自由基活性以及清除超氧阴离子自由基活性4 种不同的分析方法,检测分析3 种不同花色种类白茶茶汤的抗氧化活性;同时测定不同花色种类白茶茶汤中的主要品质化学成分的含量,并在此基础上进行相关性分析。结果：3 种不同花色种类的白茶相比,白毫银针的抗氧化活性最强,显著高于白牡丹和寿眉的抗氧化活性（P＜0.05）;而白牡丹和寿眉的抗氧化活性之间不存在显著性差异（P＞0.05）。游离氨基酸、莽草酸、表没食子儿茶素没食子酸酯等化学成分的含量在3 种不同花色种类的白茶茶汤中存在显著差异（P＜0.05）。相关性分析表明,白茶茶汤的抗氧化活性主要与儿茶素类化合物总量、总黄酮含量以及有机酸总量具有较大的相关性,其中与表没食子儿茶素没食子酸酯和奎尼酸的相关性最强。结论：不同花色种类白茶的抗氧化能力及其品质化学成分的含量水平存在较大差异。     

3 种食用菌多糖自由基清除作用研究

研究金顶侧耳、姬菇和毛头鬼伞3 种食用菌粗多糖和精多糖对超氧阴离子自由基(O2·)、羟自由基(·OH)、DPPH 自由基的清除作用。结果表明,金顶侧耳、姬菇和毛头鬼伞精多糖与粗多糖相比,对O2·清除率分别提高62.5%、58.3% 和51.5%;对·OH 清除率分别提高33.1%、42.3% 和33.2%;对DPPH 自由基清除率分别提高56.0%、63.5% 和60.3%。粗多糖经精制后,自由基清除能力明显提高,表明多糖是清除自由基的主要功效成分。     

Antioxidant Effects of Herbal Tea Leaves from Yacon (Smallanthus sonchifolius) on Multiple Free Radical and Reducing Power Assays,Especially on Different Superoxide Anion Radical Generation Systems

Yacon (Smallanthus sonchifolius), a native Andean plant, has been cultivated as a crop and locally used as a traditional folk medicine for the people suffering from diabetes and digestive/renal disorders. However, the medicinal properties of this plant and its processed foods have not been completely established. This study investigates the potent antioxidative effects of herbal tea leaves from yacon in different free radical models and a ferric reducing model. A hot‐water extract exhibited the highest yield of total polyphenol and scavenging effect on 1,1‐diphenyl‐2‐picryl hydrazyl (DPPH) radical among four extracts prepared with hot water, methanol, ethanol, and ethylacetate. In addition, a higher reducing power of the hot‐water extract was similarly demonstrated among these extracts. Varying concentrations of the hot‐water extract resulted in different scavenging activities in four synthetic free radical models: DPPH radical (EC₅₀ 28.1 μg/mL), 2,2′‐azinobis(3‐ethylbenzothiazoline‐6‐sulfonic acid) cation radical (EC₅₀ 23.7 μg/mL), galvinoxyl radical (EC₅₀ 3.06 μg/mL), and chlorpromazine cation radical (EC₅₀ 475 μg/mL). The yacon tea‐leaf extract further demonstrated superoxide anion (O₂^?) radical scavenging effects in the phenazine methosulfate‐NADH‐nitroblue tetrazolium (EC₅₀ 64.5 μg/mL) and xanthine oxidase assay systems (EC₅₀ 20.7 μg/mL). Subsequently, incubating human neutrophilic cells in the presence of the tea‐leaf extract could suppress the cellular O₂^? radical generation (IC₅₀ 65.7 μg/mL) in a phorbol 12‐myristate 13‐acetate‐activated cell model. These results support yacon tea leaves may be a good source of natural antioxidants for preventing O₂^? radical‐mediated disorders.     

Comparative Flavonoids Contents of Selected Herbs and Associations of Their Radical Scavenging Activity with Antiproliferative Actions in V79-4 Cells

ABSTRACT:  To elucidate the health benefit of herbal teas on the cytotoxicity induced by H₂O₂ in V79-4 cells, herbal extracts and its flavonoids were tested using lactate dehydrogenase release and determining intracellular reactive oxygen species generation and antioxidant activity with superoxide radical scavenging assay. Significant decrease in cell viability was observed on V79-4 cells treated with H₂O₂ (1 mM), while herbal extracts and its flavonoids including catechin and epigallocatechin gallate prevented the LDH release from H₂O₂ cytotoxicity. Total catechin contents of green tea (65.6 mg/g of dry matter) were significantly higher than other herbal teas (35.8 to 1.2 mg/g of DM). The relative concentration of the 4 major tea catechins ranked EGCG > EGC > EC > C. Green tea exhibited the lowest IC₅₀ values (2 g fresh herb/100 mL) of superoxide radical scavenging activity among the tested herbal tea, which indicates powerful antioxidant activity in O₂^·− radicals scavenging, followed by black tea, dandelion, hawthorn, rose hip, chamomile.     

不同贮藏年份蜜兰香型岭头单丛茶抗氧化成分及抗氧化活性

测定了贮藏5、10、15、20年和2016年制作(实验当年做为对照)的蜜兰香型岭头单丛茶中茶多酚、黄酮、游离氨基酸、茶黄素、儿茶素和咖啡碱等几种抗氧化成分的含量及其抗氧化活性,并研究了两者之间的相关性。结果表明:在20年贮期内,随着贮藏时间的延长,蜜兰香型岭头单丛茶的茶多酚、黄酮和儿茶素含量上升,游离氨基酸和咖啡碱含量下降,茶黄素含量先上升后下降。不同贮藏年份的岭头单丛茶清除DPPH自由基和羟自由基的能力,抑制超氧阴离子自由基的能力及还原力均有差异,通过综合指数(APC)评价抗氧化能力结果可知:抗氧化活性贮藏20年>10年>15年>5年>对照。单丛茶水提物对DPPH自由基和羟基自由基的清除能力与茶多酚、黄酮、儿茶素的含量呈显著性正相关(p<0.05)。还原能力和抑制超氧阴离子自由基能力与各抗氧化成分之间相关性不显著(p>0.05)。     

Proximate composition and free radical scavenging activity of edible fruits from the Argentinian Yungas

The proximate composition and free radical scavenging effect of native food plants gathered in the Argentinian Yungas have been assessed. Some 25 samples were collected for proximate analysis and free radical scavenging effect of their MeOH‐soluble extracts. Total acidity, phenolics and solid content of 16 preserves prepared from native fruits have been determined. The samples belong to 13 different species corresponding to eight plant families, mainly Myrtaceae, Solanaceae and Ulmaceae. The highest organic acid contents (as citric acid) were found in the preserves of Psidium guineense and Cyphomandra betaceae with the lowest in Sideroxylon obtusifolium and Myrciantes pungens. Total phenolics in the preserves ranged from 0.34 g kg^?1 DM in Rhipsalis flocosa to 7.30 g kg^?1 DM in Celtis iguanae. The MeOH‐solubles of the fruits/petioles as well as the preserves were assessed for inhibition of the enzyme xanthine oxidase (XO), decoloration of the free radical 2,2‐diphenyl‐1‐picryl‐hydrazyl (DPPH) and scavenging of the superoxide anion. The highest effect of the solubles towards the DPPH assay was observed for M pungens and Rubus imperialis before processing. None of the samples was effective as a XO inhibitor or superoxide anion scavenger. The best protein sources in our study were the fruits of Carica quercifolia, Celtis iguanae and Sambucus nigra. Copyright © 2005 Society of Chemical Industry     

马兰提取物抗氧化性研究

目的：考查马兰中水提取物和乙醇提取物抗氧化活性。方法：分别以水和乙醇为提取液,探讨马兰中水提取物和乙醇提取物的抗氧化活性,包括还原能力以及清除DPPH自由基、羟自由基、超氧阴离子自由基和H2O2的能力。 结果：提取物主要成分为黄酮类化合物。马兰水提取物的抗氧化活性为：超氧阴离子自由基(IC50 2.2μg/mL)＞H2O2(IC50 2.8μg/mL)＞羟自由基(IC50 5.6μg/mL)＞DPPH自由基(IC50 8.5μg/mL) ＞金属螯合性(IC50 28.0μg/mL)。而马兰乙醇提取物的抗氧化活性为：H2O2(IC50 2.0μg/mL) ＞超氧阴离子自由基(IC50 2.1μg/mL)>羟自由基(IC50 2.5μg/mL)> DPPH自由基(IC50 5.6μg/mL) ＞金属螯合性(IC50 51.2μg/mL)。结论：马兰提取物具有较强的抗氧化活性。     

海芦笋提取物体外抗氧化活性的研究

通过测定海芦笋水提物、醇提物、海芦笋粗多糖和精多糖还原能力及对羟自由基(·OH)和超氧阴离子自由基(O2·)的清除能力,研究海芦笋提取物的抗氧化活性。结果表明：海芦笋水提物、醇提物和多糖对·OH 都有较强清除作用,但清除能力均低于对照物VC;在相同浓度下,各提取物的还原能力和对·OH 的清除能力呈相同趋势,均是海芦笋水提物较醇提物强,粗多糖较精多糖强。但海芦笋水提物和多糖在实验所选浓度范围内,对邻苯三酚自氧化没有抑制作用,而醇提物显示出一定抑制活性。     

板栗壳棕色素对活性氧自由基的清除作用

从清除羟基自由基和超氧阴离子自由基、抗脂质过氧化以及还原能力等方面对板栗壳棕色素的抗活性氧自由基进行了试验研究和评价,结果表明:板栗壳棕色素对 Fenton 反应产生的羟基自由基和光照核黄素产生的超氧阴离子自由基均具有较强清除作用.对脂质过氧化过程中产生的脂自由基和脂过氧化自由基有明显的抑制作用.且在试验浓度范围内,其最大清除率或抑制率分别可达到 83.0%、82.1%和 88.5%.表明板栗壳棕色素是一种优良的天然抗氧化剂和自由基清除剂.     

云南种植的三种颜色玛咖体外抗氧化活性分析?

采用铁离子还原能力(FRAP)、Fenton 反应体系、DPPH 法及邻苯三酚自氧化体系评价种植于云南的玛咖乙醇提取物体外抗氧化能力。结果表明：在云南种植的3 种颜色玛咖乙醇提取物均表现出抗氧化活性,且呈现一定的量效关系,但在4 种抗氧化体系中的结果有所不同;紫色玛咖的总还原能力最强,黄色次之,白色最弱,3200m 海拔地区种植的玛咖高于其他相对低海拔地区的玛咖;紫色和黄色玛咖的清除DPPH 自由基能力高于白色,3200m 海拔地区种植的玛咖高于其他相对低海拔地区的玛咖;3 种颜色玛咖对羟自由基的清除能力明显,各样品的清除率在加入量为200μL 时均达到90% 以上,3000m 海拔地区种植的黄色玛咖最强;3 种颜色玛咖对超氧阴离子自由基的清除能力在实验范围内均较弱。     

Analysis of Free-Radical Scavenging of Yerba Mate (Ilex paraguriensis) using Electron Spin Resonance and Radical-Induced DNA Damage

ABSTRACT:  Mate (MT) is a popular South American beverage that has been used as a traditional medicine for centuries, spurring recent interest in its nutraceutical properties. MT is prepared as an infusion of leaves from the Yerba Mate ( llex paraguriensis ) tree. MT has been reported to have antioxidant properties  in vitro  and  in vivo , but these have not been fully characterized in terms of effects against specific radicals. Accordingly, we examined the antioxidant effects of an MT infusion against hydroxyl and superoxide radicals in both chemical and cell culture assays. MT infusions were prepared at 3.10 g/L in boiling water and diluted to experimental dilutions from this stock. Electron spin resonance (ESR) experiments indicated that MT scavenged hydroxyl radicals (produced via the Fenton reaction) and superoxide radicals (produced via the xanthine/xanthine oxidase enzymatic reaction) at all concentrations tested ( P  < 0.05). Further controls indicated that superoxide radical scavenging was not due to xanthine oxidase inhibition. MT scavenged hydroxyl radicals and decreased cellular oxygen consumption in a dose-dependent manner in Cr(VI)-stimulated RAW 264.7 cells, based on ESR and oxygraph measurements ( P  < 0.05). Similarly, MT also inhibited hydroxyl-radical-induced lipid peroxidation and DNA damage in a dose-dependent manner in RAW 264.7 cells, based on malondialdehyde and Comet assay data ( P  < 0.05). This study indicates that MT possesses potent antioxidant effects against hydroxyl and superoxide radicals in both chemical and cell culture systems, as well as DNA-protective properties. These data further clarify the reported antioxidant effects of Yerba Mate infusions.     

Antioxidative capacity of rhizome extract and rhizome knot extract of edible lotus (Nelumbo nuficera)

Extracts from rhizomes (Lotus roots) knot (LRK) and whole rhizomes (LR) were investigated for their antioxidative capacity in comparison with commonly used antioxidants from plant material. Radical scavenging activity was measured spectrophotometrically using the stable radicals 1-diphenyl-2-picrylhydrazyl and 2,2′-azino(3-ethylbenzothioazolino-6-sulfonate), and measured by electron spin resonance (ESR) trapping of the transient carbon-centered 1-hydroxyethyl radical (generated in a Fenton-type reaction). Efficiencies as chain-breaking antioxidants were evaluated by electrochemical measurement of oxygen consumption rate in a peroxidating methyl linoleate emulsion. LRK exhibited high antioxidative capacity, as measured by each of four different methods. LR, however, only showed a significantly high scavenging activity for small carbon-centered radicals, as measured by the ESR method. Total phenol content in the plant extract correlated with the antioxidant capacity, except for the scavenging of carbon-centered radical. Lotus rhizomes knot, as a waste from food production, will be a potential material for extracting antioxidants.