Extraction Storage and Qualitative Analysis of Free Radicals Scavenging Substances from Sweet Potato Leaves

[ Objective ] The research aimed to provide reference for the application of extracts from sweet potato leaves in anti-aging cosmetics. [ Method ] The extraction and storage conditions for free radicals scavenging substances from sweet potato leaves were optimized by orthogonal test and the bioactive components in extracts were investigated by correlation analysis. [ Result] Sweet potato leaves contain the bioactive substances scavenging DPPH free radical and hydroxyl free radical. Extracting solvent species is the most important factor that influencing extraction yield. The optimal extraction and storage conditions are as following： water as solvent, pH 8.0 of extracting liquid, storage at 25 ℃. There is a good positive linear relationship between the content of total phenols in sweet potato leaves and corresponding scavenging rates against both the DPPH free radical and hydroxyl free radical. For the content of total flavones in sweet potato leaves, just a correlation with scavenging rate against hydroxyl free radical shown in test. [ Conclusion] The phenols in ex- tracts could effectively scavenge both the DPPH free radical and hydroxyl free radical, whereas the flavones in extracts can only function on the hydroxyl free radical.

Component Analysis and Free Radicals Scavenging Activity of Physalis alkekengi L. Polysaccharide

A crude polysaccharide was extracted from Physalis alkekengi L. fruit. HPLC was used for the component analysis of the polysaccharide. The results indicate that Physalis alkekengi L. polysaccharide（PAP） was composed of rhamnose, xylose, arabinose, galactose, and glucose. Free radicals scavenging activity of PAP was studied through 3 free radicals scavenging tests. PAP exhibited high scavenging effects on OH and DPPH radicals, and both the scavenging rates were about 80%. The scavenging rate of O2^- radical was about 22%.

STUDIES ON FREE RADICAL SCAVENGING ACTIVITY IN CHINESE SEAWEEDS PART I. SCREENING RESULTS

Antioxidants have attracted the attention of researchers due to their beneficial effects as free radical scavengers. Application of a stable free radical named 1,1 diphenyl 2 picrylhydrazyl(DPPH) to screen the free radical scavenging activity in 27 species of Chinese seaweed showed that 15 of them had significant activity in at least one of the organic solvent extracts. The most interesting seaweed species were Gelidium amansii, Gloiosiphonia capillaris, Polysiphonia urceolata, Sargassum kjellmanianum, Desmarestia viridis, and Rhodomela teres.

Phytochemical screening and free radical scavenging activity of Citrullus colocynthis seeds extracts

Objective:To study the phytochemical screening of different extracts from Citrullus colocynthis(C.colocynthis)seeds extracts and to assess their antioxidant activity on the DPPH free radical scavenging.Methods:Phytochemical screening,total content of polyphenols and flavonoids of C.colocynthis seeds extracts,including a crude aqueous extract(E1),a defatted aqueous extract(E2),a hydromethanolic extract(HM),an ethyl acetate extract(EA)and a n-butanol extract(n-B)was carried out according to the standard methods and to assess their corresponding effect on the antioxidant activity of this plant.Results:None of these extracts contained detectable amount of alkaloid,quinone,antraquinone,or reducing sugar.Catechic tannins and flavonoids were abundant in E1,MM and EA,whilst terpenoids were abundantly present in E1 and n-B but only weekly in HM.Coumarins were found in E2,EA and n-B.Polyphenols,expressed as gallic acid equivalent,amounted,per 100 g plant matter,to 329,1002 and 150 mg in EA,HM an E1 respectively.Flavonoids,expressed as catechin equivalent,amounted.per 100 g plant matter to 620,241 and 94 mg in EA,HM and E1 respectively.Comparable values were found in n-B and E1,with lower values in E2.Quercetin,myricetin and gallic acid were found in the EA and HM extracts by thin layer chromatography,The antioxidative effect of these extracts yielded,when tested at a concentration of 2000μg/mL in a 1,1-diphenyl-2-picrylhydrazyl assay,a reducing percentage of 88.8%with EA,74.5%with HM and 66.2%with E1,and corresponding IC_(50)of 350,580and 500μg/mL as compared to 1.1μg/mL for ascorbic acid.Conclusions:These qualitative and quantitative analytical data document the presence in C.colocynthis extracts of such chemical compounds as flavonoids responsible for the antioxidant activity,as well as other biological activities of this plant.

Evaluation of free radical scavenging activity of various extracts of leaves from Kedrostis foetidissima(Jacq.)Cogn

The present study was aimed to investigate the in vitro free radical scavenging activity of various leaf extracts(aqueous,methanol,acetone chloroform and petroleum ether)of Kedrostis foetidissima.In vitro free radical scavenging activities of the extracts were assessed against DPPH and hydroxyl radicals.The metal chelating activity and reducing power ability of the extracts were also determined.The free radical scavenging activity was found to be high in methanolic extract for DPPH and hydroxyl radicals in a concentration dependent manner followed by chloroform,aqueous,acetone and petroleum ether extracts.The metal chelating activity and reducing power ability was also found to be high in methanolic extract.The difference in scavenging potential of the extracts may be due to variation in the percentage of phytoconstituents extracted in various solvents.Thus the result suggests that the methanolic leaf extract of K.foetidissima could serve as a potential source of antioxidants and can be explored as a therapeutic agent in free radical induced diseases.©2015 Beijing Academy of Food Sciences.Production and hosting by Elsevier B.V.All rights reserved.

Butein imparts free radical scavenging, anti-oxidative and proapoptotic properties in the flower extracts of Butea monosperma

The flower of Butea monosperma(Lam.)(Fabaceae)has been used in traditional Indian medicine in the treatment of many ailments including liver disorders.To understand the pharmacological basis of its beneficial effects,the extracts of dried flowers in water,methanol,butanol,ethyl acetate and acetone were evaluated for free radical scavenging and pro-apoptotic activities in cell cultures(human hepatoma Huh-7 cell line and immortalized AML-12 mouse hepatocytes).Butrin and butein-the active constituents of flower extracts-were used as reference molecules.The levels of cell injury markers like lactate dehydrogenase,glutathione and lipid peroxidation and primary antioxidant enzymes glutathione S-transferase and catalase were also measured.The aqueous and butanolic extracts exhibited better 2,2-diphenyl-1-picrylhydrazyl scavenging and cytotoxic activities in hepatoma cells than in immortalized hepatocytes.Interestingly,butein inhibited 2,2-diphenyl-1-picrylhydrazyl radical better than butrin.The aqueous and butanolic extracts were further investigated for hepatoprotection against carbon tertrachloride-induced biochemical changes and cell death.Both extracts,just as butrin and butein,significantly reversed the cellular glutathione levels and lipid peroxidation,and glutathione–S-transferase activity.Lactate dehydrogenase leakage and cell death were also prevented.However,only butein revived the catalase activity.Thus,the butein content of Butea monosperma flower extracts is important for free radical scavenging activity,apoptotic cell death and protection against oxidative injury in hepatic cells.

Study on the Free Radical Scavenging Activity of Sea Cucumber (Paracaudina chinens var.) Gelatin Hydrolysate

Gelatin from the sea cucumber(Paracaudina chinens var.) was hydrolyzed by bromelain and the hydrolysate was found to have a high free radical scavenging activity. The hydrolysate was fractionated through an ultrafiltration membrane with 5 kDa molecular weight cutoff(MWCO). The portion(less than 5 kDa) was further separated by Sephadex G-25. The active peak was col-lected and assayed for free radical scavenging activity. The scavenging rates for superoxide anion radicals(O2·-) and hydroxyl radi-cals(·OH) of the fraction with the highest activity were 29.02% and 75.41%,respectively. A rabbit liver mitochondrial free radical damage model was adopted to study the free radical scavenging activity of the fraction. The results showed that the sea cucumber gelatin hydrolysate can prevent the damage of rabbit liver and mitochondria.

Sonme Factors that Affect the Free Radical-scavenging Activity of Tea Extracts

Some factors that affect the free radical-scavenging activety of two tea extracts were studied in vitro. It was found that concentration of tea extract or heating tea extract or treating with activated carbon and diatomite all had obvious effect on the scavenging activety of green tea extract ,but heating or treating with diaomite had less effect on the scavenging activity of black tea extract. Ascorbic acid, for having synergic effect with tea extracts, could enhance the scavenging activity of tea extracts markedly, and the contrary was cupric ion. Reducing sugars such as fructose and glucose also had some syncrgic effect to tea extracts.

Effects of Chuanxiongqin hydrochloride on increasing the fluidity of brain cell membrane and scavenging free radicals in model rats with ischemia/reperfusion injury

BACKGROUND: The fluidity of cell membrane can be affected by various factors. Many experiments have confirmed that the ischemia/reperfusion of organic tissue can increase the contents of free radicals, which lead to high rigidity and low fluidity of cell membrane, and the conditions can be changed by Chuanxiongqin. OBJECTIVE: To observe the effect and mechanism of Chuanxiongqin hydrochloride on the fluidity of brain cell membrane in rat models of ischemia/reperfusion. DESIGN: A completely randomized controlled animal trial. SETTINGS: Institute of Brain Sciences; Department of Physiology, Medical College, Datong University. MATERIALS: Twenty male grade Ⅰ Wistar rats of 170-220 g were randomly divided into model group (n =10) and control group (n =10). Chuanxiongqin hydrochloride (molecular mass was 172.2) was purchased from the National Institute for the Control of Pharmaceutical and Biological Products (batch number: 0817-9803); Spin labelers: 5-doxyl-stearlic acid methylester (5DS), 16-doxyl-stearlic acid methylester (16DS), xanthine, xanthine oxidase (XOD) and 5,5-dimeth-1-pyrroline- N-oxide (DMPO) from Sigma Company; Bruker ESP 300 electron paramagnetic resonance (EPR) spectrometer by Bruker Company (Germany). METHODS: The experiments were carried out in the State Key Laboratory of Natural and Biomimetic Drugs, Peking University from June 2001 to July 2002. In the model group, rats were made into models of cerebral ischemia by 30-minute ligation and 2-hour reperfusion of common carotid arteries; The rats in the control group were not made into models. The order parameter (S) and rotational correlation time (τc) were detected with the ESR spectrometer by means of spin labeling. The greater the S and τc, the smaller the fluidity. Meanwhile, the clearance rate of free radicals was detected with ESR spin trapping. The measurement data were compared using the t test. MAIN OUTCOME MEASURES: The S, τc and clearance rates of O2 · and OH· free radicals were compared between the model group and control group. RESULTS: The S and τc in the model group [0.738 4±0.003 5; (8.472±0.027)×10-10 s/circle] were obviously different from those in the control group [0.683 9±0.008 3; (7.945±0.082)×10-10 s/circle, t =5.731, 5.918, P < 0.05], which suggested that ischemia/reperfusion injury decreased the fluidity of brain cell membrane. After adding Chuanxiongqin hydrochloride, there were no obvious differences between the model group [0.688 5±0.030 5; (7.886±0.341)×10-10 s/circle] and control group (P > 0.05), indicating that Chuanxiongqin hydrochloride could recover the fluidity of brain cell membrane after ischemia/reperfusion injury close to the level in the normal control group. Chuanxiongqin hydrochloride could directly scavenge the O2 · and OH· free radicals, and the maximal clearance rates were 83.92% and 44.99% respectively. CONCLUSION: Chuanxiongqin hydrochloride increases the fluidity of membrane of ischemia-injured brain cell by scavenging both O2 ·and OH· free radicals.

Comparative Analysis of the Antioxidant and Free-Radical Scavenging Activities of Different Water-Soluble Extracts of Green, Black and Oolong Tea Samples

Extracts of 40 green, black or oolong high quality tea samples from China, Japan or India among others, have been prepared according to the manufacturer instructions and assayed for total polyphenols content using the Folin-Ciocalteu phenol reagent. In addition, the antioxidant activity was assessed by the CUPRAC method and the free-radical scavenging activity was determined using the antioxidant-promoting decay of the stable free-radical DPPH. Caffeine and the most important catechins were identified using RP-HPLC previously validated method. Results from this study suggest that teas, even though they content similar amounts of caffeine, differ considerably in polyphenolic content and therefore in antioxidant and free-radical scavenging activities, depending on the part of the plant used and the process applied to the material for the preparation of the final tea presentation. Correlations among Folin-Ciocalteau’s reactivity of tea samples, the neocuproine reactivity, the DPPH decolourization and the HPLC analysis suggest that the antioxidant activity is due essentially to polyphenolic compounds present in teas, mainly EGCG. The best extraction method of tea powder was the one indicated by dealers. In general, the richest samples belong to green teas and the poorest samples belong to the black teas.

Evaluation of lipid peroxidation inhibition and free radical scavenging abilities of 5,6,7-trimethoxy dihydroflavonols

Four naturally rare 5,6,7-trimethoxy-2,3-cis-dihydroflavonols （3-6） and two 5,6,7-trimethoxy-2,3-trans-dihydroflavonols （7-8） were designed and synthesized. Their antioxidative properties were evaluated by way of examining their scavenging capacities towards DPPH and O2^＊- free radicals, as well as by measuring their inhibitory ability against LPO. Both the 2,3-trans and the 2,3- cis conformers exhibited certain quenching abilities to DPPH and O2^＊- radicals, while most of the synthetic dihydroflavonols demonstrated remarkable inhibition to LPO.

Why Static O-H Bond Parameters Cannot Characterize the Free Radical Scavenging Activity of Phenolic Antioxidants: ab initio Study

The static O-H bond parameters including O-H bond length, O-H charge difference, O-H Mulliken population and O-H bond stretching force constant (k) for 17 phenols were calculated by ab initio method HF/6-31G**. In combination with the O-H bond dissociation enthalpies (BDE) of the phenols determined by experiment, it was found that there were poor correlationships between the static O-H bond parameters and O-H BDE. Considering the good correlationship bt tween O-H BDE and logarithm of free radical scavenging rate constant for phenolic antioxidant, it is reasonable to believe that the ineffectiveness of static O-H bond parameters in characterizing antioxidant activity arises from the fact that they cannot measure the O-H BDE.

Free Radical-Scavenging Properties and Antioxidant Activity of Fractions from Cranberry Products

Lipid peroxidation inhibition capacity and antiradical activity were evaluated in HPLC fractions of different polarity obtained from two cranberry juices and three extracts isolated from frozen cranberries and pomace containing antho-cyanins, water-soluble and apolar phenolic compounds, respectively. Compounds with close polarities were collected to obtain between three and four fractions from each juice or extract. The cranberry phenols are good free radi-cal-scavengers, but they were less efficient at inhibiting the lipid peroxidation. Of all the samples tested, the intermediate polarity fraction of extract rich in apolar phenolic compounds of fruit presented the highest antiradical activity while the most hydrophobic fractions of the anthocyanin-rich extract from fruit and pomace appeared to be the most efficient at inhibiting the lipid peroxidation. The antioxidant or pro-oxidant activity of fractions increased with the con-centration. The phenol polarity and the technological process to manufacture cranberry juice can influence the antioxidant and antiradical activities of fractions.

Application of Theories of Free Radicals and Electron Spin Resonance for Research of Traditional Chinese Medicine

The present paper reviews new findings in redoxproperties of the active constituent of Chinese herbalmedicine(CHM),a kind of CHM or a compoundprescription as an antioxidant.Firstly,we have studiedtheir antioxidant and prooxidant actions with electronspin resonance(ESR).The results show that the activecomponents from over 10 kinds of CHM are able toscavenge the oxygen free radicals but propyl gallate

Tentative Identification of Phytochemicals and Antioxidant Activities during Fruit-Ripening on Chamaedorea radicalis Mart.

This work aims to determine the phytochemical characterization of the pericarp of Chamaedorea radicalis Mart.fruit as a non-timber product with potential to obtain phytochemicals with potential applications in the industry.Fruit from C.radicalis were grouped in four ripening stages named as S1,S2,S3 and S4,according to maturity;S1 the most unripe stage and S4 the completely ripe stage.Determinations of total phenolic compounds,free radical scavenging activities and total flavonoid contents using spectrophotometric methods were done.Also,the tentative identification of phytochemicals during fruit ripening was done using UPLC-MS-MS.Total phenolic compound(TPC)content ranged from 7.24 to 12.53 mg gallic acid equivalents per gram of fresh weight(mg GAE/g FW).Total flavonoids(TF)contents ranged from 0.163 to 0.23 mg of quercetin equivalents per g FW(mg QE/g FW).Free radical scavenging activity against DPPH and ABTS radicals varied from 40.80 to 53.68 and from 22.29 to 37.76 mmol Trolox equivalents g FW(mmol TE/g FW),respectively.Antioxidant assay in vitro by FRAP(ferric reducing antioxidant power)method showed that S3 was the highest level with antioxidant power while S4 was the lowest with Red ripeness stage showed the lowest contents for all determinations.Mass spectrometry allowed detection of 26 compounds,including phenolics,alkaloids and saponins.Afzelin,Kaempferol 3-neohesperidoside and the four saponins identified were present in all ripeness stages.Preliminary phytochemical identification and the spectrophotometric determinations showed that the pericarp of C.radicalis presented antioxidants and compounds related to alkaloids,phenolics and saponins.The presence and abundance of each phytochemical regarding each ripeness stage should be considered.

桑葚果实、桑葚干酒和桑葚利口酒中酚类化合物的高效液相色谱串联电喷雾二级质谱分析及其抗氧化活性比较

为了解桑葚果酒和桑葚果实在酚类化合物的种类、含量及抗氧化活性之间的差异,该研究以桑葚果实、桑葚干酒和桑葚利口酒为试材,采用高效液相色谱串联电喷雾二级质谱(high performance liquid chromatography-electron spray ionization-mass spectrometry/mass spectrometry,HPLC-ESI-MS/MS)对酚类化合物进行定性定量分析。HPLC-ESI-MS/MS分析结果表明,在桑葚果实、桑葚干酒和桑葚利口酒中均检测到花青素-3-葡萄糖苷和花青素-3-芸香糖苷,而花葵素-3-芸香糖苷和4-丙酮酸-花青素-3-葡萄糖苷分别只在桑葚干酒和桑葚利口酒中被检测到。同时,桑葚干酒和桑葚利口酒中花青素-3-葡萄糖苷和花青素-3-芸香糖苷的含量均显著低于桑葚果实。此外,在桑葚果实中鉴定的8种非花色苷多酚中,仅有4种存在于桑葚干酒和桑葚利口酒中。相较于桑葚果实,桑葚干酒和利口酒中原儿茶素、二氢槲皮素和槲皮素的含量显著升高。桑葚果实DPPH自由基清除能力介于桑葚干酒和利口酒之间。综上,桑葚果酒和桑葚果实中酚类化合物的种类、含量及抗氧化活性之间差异显著。

南、北五味子蛋白抗氧化活性和对HepG2细胞氧化应激损伤的修复作用

采用碱提酸沉法提取南五味子蛋白(Schisandra sphenanthera protein,SSP)和北五味子蛋白(Schisandra chinensis protein,SCP),并测定其体外清除自由基能力、还原能力和对H_(2)O_(2)诱导的HepG2细胞氧化损伤的修复作用。采用CCK-8法检测细胞增殖率,荧光检测胞内活性氧水平,酶联免疫吸附法检测细胞氧化应激产物和抗氧化物酶系活性。结果表明,SSP清除自由基和还原能力均优于SCP,SSP与SCP都能对H_(2)O_(2)诱导HepG2造成的细胞生长抑制起到修复作用(P<0.05),显著降低胞内活性氧、丙二醛水平(P<0.05),显著提高超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶和谷胱甘肽的水平(P<0.05),且SSP对H_(2)O_(2)诱导的HepG2氧化应激损伤修复能力优于SCP。综上,2种五味子蛋白中SSP更适合应用于抗氧化功能性食品或保健食品工业中。

体外模拟胃肠消化过程中云南金花茶抗氧化能力变化

为探究云南金花茶在消化过程中总黄酮、总多酚释放及功能活性的变化情况,该研究采用体外模拟胃肠消化模型测定了云南金花茶消化液中总黄酮、总多酚浓度及自由基清除能力、铁离子还原能力和脂质过氧化抑制能力,并对其进行了相关性分析。结果表明,模拟胃消化过程中,云南金花茶总黄酮、总多酚含量均比口腔消化高,最高分别达229.99、327.65µg/mL,经肠消化后,总黄酮含量在38.56~78.88µg/mL之间,总多酚含量在30.04~65.52µg/mL之间,含量较低。在功能活性测定中,较口腔消化,胃消化阶段的DPPH·、ABTS+·清除能力和铁离子还原能力显著升高,脂质过氧化抑制能力随消化的进行呈上下波动;经肠消化后,DPPH·、ABTS+·清除能力和铁离子还原能力急剧下降,且均比胃消化阶段低,而脂质过氧化抑制能力则趋于平稳或逐渐上升。通过相关性及主成分分析,各指标之间存在一定的关联性。综上所述,云南金花茶是较好的活性物质来源,该研究可为探索云南金花茶在人体消化过程中活性物质的变化及作用机制提供一定的实验基础和新思路。

红曲糟源酶解蛋白肽的功能性评价

【目的】研究不同蛋白酶对红曲糟酶解的效果,并对其酶解液进行功能性评价,为红曲糟源蛋白肽的研发制备提供理论支持。【方法】以提高蛋白含量后的红曲糟为原料,利用碱性蛋白酶、胰蛋白酶、动物蛋白酶、木瓜蛋白酶、菠萝蛋白酶、胃蛋白酶、复配酶制剂F106、酵母抽提酶等不同蛋白酶进行酶解,考察其酶解率、1,1-二苯基-2-三硝基苯肼(DPPH)和2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)自由基清除抗氧化能力、黄嘌呤氧化酶(Xanthine oxidase,XOD)和血管紧张素转化酶(Angiotensin converting enzyme,ACE)抑制活性等生物活性功能。【结果】蛋白酶解率最高的为动物蛋白酶和胰蛋白酶酶解处理,分别为(71.43±1.03)%和(70.20±0.32)%。DPPH自由基清除抗氧化能力最优的是胃蛋白酶、碱性蛋白酶和酵母抽提酶酶解处理,蛋白肽的半数效应浓度(Median effective concentration,EC50)分别为(2.78±0.34)mg·mL^(-1)、(3.02±0.03)mg·mL^(-1)、(3.24±0.65)mg·mL^(-1);ABTS自由基清除抗氧化能力、XOD抑制活性能力最优的均为胃蛋白酶和碱性蛋白酶酶解液,其蛋白肽的EC50值分别为(1.54±0.07)mg·mL^(-1)、(6.45±0.27)mg·mL^(-1)和(10.71±0.06)mg·mL^(-1)、(17.68±0.04)mg·mL^(-1),二者的XOD半数抑制指数分别为(1.28±0.01)、(1.78±0.03);ACE抑制活性最优的为碱性蛋白酶酶解液和木瓜蛋白酶酶解液,蛋白肽的EC50值均为(0.27±0.01)mg·mL^(-1),半数抑制指数分别为(118.40±3.53)、(98.35±1.95)。【结论】红曲糟源蛋白经不同蛋白酶酶解后的肽段具有不同的生物活性,其中以胃蛋白酶酶解的XOD抑制活性能力较强,碱性蛋白酶和木瓜蛋白酶酶解的ACE抑制活性较优。

塔拉多糖超声提取及其自由基清除能力研究

为研究塔拉多糖的提取工艺及抗氧化活性,选用超声辅助提取法对塔拉多糖进行提取,围绕液料比、超声功率、提取温度和提取时间展开单因素试验,并通过响应面法对影响参数进行优化。经过统计分析和数学回归模型拟合得知,决定系数(R~2)为0.956,即精确系数为95.60%,表明该模型吻合度较好,适用于塔拉多糖超声提取工艺的优化,其最佳参数为10∶1 mL/g, 20 min, 150 W,40℃,实际平均得率为24.58%,接近理论期望得率25.07%。此外,又对塔拉多糖自由基的清除能力进行一阶动力学方面的研究,以维生素C为对照,根据试验数据得知,塔拉多糖的DPPH、ABTS~+对应IC 50分别为524.07和3 496.88μg/mL,说明其在自由基清除能力方面表现较为优异,因此在抗氧化性方面具有较好的应用价值。研究成果最终可为植物多糖的提取、纯化与应用提供可靠的理论指导与数据支撑。