Radical scavenging activities of Rio Red grapefruits and Sour orange fruit extracts in different in vitro model systems

Antioxidant fractions from two different citrus species such as Rio Red (Citrus paradise Macf.) and Sour orange (Citrus aurantium L.) were extracted with five different polar solvents using Soxhlet type extractor. The total phenolic content of the extracts was determined by Folin-Ciocalteu method. Ethyl acetate extract of Rio Red and Sour orange was found to contain maximum phenolics. The dried fractions were screened for their antioxidant activity potential using in vitro model systems such as 1,1-diphenyl-2-picryl hydrazyl (DPPH), phosphomolybdenum method and nitroblue tetrazolium (NBT) reduction at different concentrations. The methanol:water (80:20) fraction of Rio Red showed the highest radical scavenging activity 42.5%, 77.8% and 92.1% at 250, 500 and 1000 ppm, respectively, while methanol:water (80:20) fraction of Sour orange showed the lowest radical scavenging activity at all the tested concentrations. All citrus fractions showed good antioxidant capacity by the formation of phosphomolybdenum complex at 200 ppm. In addition, superoxide radical scavenging activity was assayed using non-enzymatic (NADH/phenaxine methosulfate) superoxide generating system. All the extracts showed variable superoxide radical scavenging activity. Moreover, methanol:water (80:20) extract of Rio Red and methanol extract of Sour orange exhibited marked reducing power in potassium ferricyanide reduction method. The data obtained using above in vitro models clearly establish the antioxidant potential of citrus fruit extracts. However, comprehensive studies need to be conducted to ascertain the in vivo bioavailability, safety and efficacy of such extracts in experimental animals. To the best of our knowledge, this is the first report on antioxidant activity of different polar extracts from Rio Red and Sour oranges.     

Fucoxanthin as the major antioxidant in Hijikia fusiformis, a common edible seaweed

The radical scavenging activity of Japanese edible seaweeds was screened by the DPPH (1-diphenyl-2-picrylhydrazyl) assay to evaluate the DPPH radical scavenging activity in organic extracts. The fresh brown alga Hijikia fusiformis showed the strongest DPPH radical scavenging activity, followed by Undaria pinnatifida and Sargassum fulvellum. The major active compound from Hijikia fusiformis in its acetone extract was identified as fucoxanthin by 13C-NMR spectroscopy.     

桦褐孔菌提取物抗氧化活性研究

以乙酸乙酯和甲醇为提取剂,采用索氏提取法,剩余残渣采用热水浸提,最终得到桦褐孔菌不同极性提取物,对其DPPH自由基、羟基自由基及超氧阴离子自由基清除活性作用进行了研究,确定桦褐孔菌的抗氧化能力,为深入研究和开发桦褐孔菌功能性食品奠定理论基础。实验结果表明桦褐孔菌具有较好的抗氧化活性,其中乙酸乙酯提取物的DPPH自由基清除率、羟基自由基清除率和超氧阴离子自由基清除率均高于其他两组分及BHT,桦褐孔菌提取物有望成为功能性食品组分中合成抗氧化剂的天然替代品。     

蓝莓果渣提取物总酚含量及抗氧化活性研究

研究了蓝莓果渣提取物总酚含量及其抗氧化活性。分别采用水、40%乙醇及纤维素酶辅助乙醇超声提取蓝莓果渣,并用Folin-Ciocalteu试剂对3种提取物的总酚含量进行评估;并采用DPPH清除实验及O2—.清除实验对3种提取物的抗氧化活性进行研究。实验结果表明,纤维素酶辅助超声提取蓝莓果渣的总酚含量最高,可达425.36±15.21 mg GAE.100 g-1DW,远远高于水提物(169.46±9.75 mg GAE.100 g-1DW)及醇提物(218.39±12.54mg GAE.100 g-1DW)中的总酚含量。且纤维素酶辅助乙醇超声提取物对DPPH的清除能力为2.67±0.13 gVc.100 g-1DW,对O—.2的清除能力2.48±0.14 g Vc.100 g-1DW,明显好于醇提物及水提物抗氧化活性。     

花脸香蘑菌丝体提取物的体外抗氧化活性

分别测定花脸香蘑Lepista sordida发酵菌丝体不同浓度乙醇溶剂提取液在4种抗氧化模型中的抗氧化作用。结果表明, 各种菌丝体提取液对二苯基苦味酰基苯肼自由基(·DPPH)和羟自由基(·OH)均有显著的清除作用, 其清除作用随着乙醇溶剂浓度的提高而降低: 在3 g/L菌丝生药浓度时, 去离子水提取物对DPPH自由基和羟自由基的清除率分别为90.55%和81.9%; 在6 g/L菌丝生药浓度时, 50%乙醇和70%乙醇提取物对DPPH自由基的清除率分别为53.7%和45.2%, 对羟自由基的清除率分别为79.4%和78.4%。各菌丝提取物也具有极强的抗脂质过氧化能力, 在5 g/L菌丝生药浓度64 h内, 水提物、50%乙醇和70%乙醇提取物对亚油酸过氧化物的抑制率分别为100%、96%和89.2%。在一定浓度下, 各提取液对邻苯三酚自氧化也均有抑制作用。     

益智的抗氧化作用

本文探讨了益智(Alpinia oxyphylla Miquel)超临界CO2提取物及其渣的水提物、正丙醇提取物和乙酸乙酯提取物的抗氧化作用,测定了总酚含量、黄酮含量、抗氧化力、还原能力、DPPH清除率.结果表明,益智超临界CO2提取物和正丙醇提取物的总酚含量最高,均为5.53%,乙酸乙酯提取物的总酚含量为4.04%,水提物总酚含量最低,为O.89%.抗氧化力与酚含量相关(R2=0.703).四种提取物中黄酮含量顺序为:乙酸乙酯提取物(6.29%)＞丙醇提取物(5.81%)＞水提取物(4.85%)＞超临界CO2提取物(4.70%).在还原能力、清除DPPH自由基和羟自由基方面,乙酸乙酯提取物表现出了很强的抗氧化能力,呈现剂量依赖关系.     

Neuroprotection and free radical scavenging effects of <Emphasis Type="Italic">Osmanthus fragrans</Emphasis>

The ethanol extract of dried flowers Osmanthus fragrans (OFE) was assessed for free radical scavenging effects measured by the bleaching of the 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical, scavenging of the hydroxyl anion, investigation of the ferric reducing/antioxidant power (FRAP) and lipid-peroxidation inhibition in rat tissues. OFE contained a high amount of total flavonoid and polyphenol. OFE presented the effects in the metal reducing power, FRAP assay with IC₅₀ values of 0.23 μg/ml, and 7.74 μg/ml, respectively. OFE presented similar activities toward the DPPH and hydroxyl anion scavenging ability with IC₅₀ values of 10 μg/ml. OFE with IC₅₀ values between 46 and 97 μg/ml inhibited lipid peroxidation initiated by ferrous chloride in rat brain, liver, heart and kidney mitochodrias. Moreover, the neuroprotective activity of OFE was investigated under different insults (glutamate, arachidonic acid, and 6-hydroxydopamine) in Wistar rat primary cortical neurons. OFE with EC₅₀ values between 66 and 165 μg/ml attenuated the neurotoxicity on MTT and LDH assays. In addition, the AKT protein expression of excitotoxicity and oxidative stress was displayed by western blotting analysis. OFE could up-regulate the glutamate and 6-OHDA decreased AKT expression. This is the first demonstration of the neuroprotective, free radical scavenging and anti-oxidative effects of O. fragrans.     

印度块菌提取物抗氧化活性的研究

对印度块菌Tuber indicum子实体的提取物,包括55%乙醇粗提物(ECE)、石油醚提取物(PEF)和乙酸乙酯提取物(EAF)的清除DPPH自由基和羟基自由基能力、铁离子鳌合能力以及各提取物的总酚含量等进行了研究和测定,结果显示,3种提取物的清除自由基能力和铁离子鳌合能力具有显著差异(P0.05);ECE对DPPH自由基的清除活性最高,其EC50值为1.61g/L;EAF对羟基自由基及铁离子表现出较强的清除或螯合的能力,其EC50值分别为3.31g/L和0.70g/L;EAF的总酚含量(2.964mg GAE/g提取物)最高,其次是ECE,总酚含量为(2.618mg GAE/g提取物);PEF的清除自由基和铁离子鳌合能力较差,其总酚含量也最低(1.124mg GAE/g提取物);总酚含量与印度块菌提取物清除自由基以及鳌合铁离子的能力密切相关。     

库克诺你果汁提取物体外清除自由基及抗氧化活性研究

本文对诺你果汁多糖、乙醇溶出物和乙酸乙酯萃取物体外对超氧阴离子(O2·)、羟自由基(·OH)、DPPH和脂质过氧化(LPO)的抑制作用进行了研究。超氧阴离子(O2·)由邻苯三酚自氧化产生;羟自由基(·OH)由Fenton反应产生;利用Fe2 诱发卵黄脂蛋白产生丙二醛(MDA),TBA法测定。所有测定均为分光光度法。结果表明,与已知抗氧化剂L抗坏血酸相比,乙醇溶出物和乙酸乙酯萃取物均有明显的捕捉自由基和抗氧化能力,而多糖捕捉自由基和抗氧化能力很低,且对O2·没有抑制作用,反而会增加其生成速度。     

余甘子活性成分含量与抗氧化性研究

研究了不同产地余甘子的总多酚、黄酮和多糖含量,同时测定了各产地余甘子提取物的抗氧化活性。结果表明,广东野生品种鲜果的总多酚含量、总黄酮含量和水溶性多糖含量最高,它们在每克干果中的含量分别为(204.15±6.85)mg没食子酸当量、(81.11±8.67)mg芦丁当量和(19.78±1.22)mg葡萄糖当量。广东野生品种提取物的抗氧化活性较高,其还原力为1.344±0.14,羟基自由基和超氧阴离子自由基的清除力分别为91.50%±3.53%,92.31%±1.30%。广东栽培种提取物的DPPH自由基清除力最高,达到92.09%±1.52%。     

Evaluation of in-vitro antioxidant and antibacterial properties of Commelina nudiflora L. extracts prepared by different polar solvents

The study explored on the commonly available weed plant Commelina nudiflora which has potential in-vitro antioxidant and antimicrobial activity. The different polar solvents such as ethanol, chloroform, dichloromethane, hexane and aqueous were used for the soxhlet extraction. The extracts were identified pharmacologically as important bioactive compounds and their potential free radical scavenging activities, and antimicrobial properties were studied. C. nudiflora extracts were monitored on their in-vitro antioxidant ability by DPPH and ABTS radical scavenging assay. Aqueous extract shows significant free radical scavenging activity of 63.4 mg/GAE and 49.10 mg/g in DPPH and ABTS respectively. Furthermore, the aqueous crude extract was used in antibacterial studies, which shows the highest inhibitory activity against Pseudomonas aeruginosa, Escherichia coli and Salmonella typhi. Among all the extracts, aqueous extract of C. nudiflora has significant control over free radical scavenging activity and inhibition of the growth of food pathogenic bacteria. Also, the aqueous extract contains abundance of phenolics and flavonoids higher than other extracts. This study explored weed plant C. nudiflora as a potential source of antioxidant and antibacterial efficacy and identified various therapeutic value bioactive compounds from GC–MS analysis.Abbreviations: ABTS, 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulphonic acid), DPPH, 2,2-diphenyl-1-picrylhydrazyl, GAE, gallic acid equivalent, GC–MS, gas chromatography and mass spectrometry     

Antioxidant activities of enzymatic extracts from brown seaweeds

Potential antioxidative activities of enzymatic extracts from seven species of brown seaweeds were evaluated using four different reactive oxygen species (ROS) scavenging assays containing DPPH (1,1-diphenyl-2-pricrylhydrazyl) free radical, superoxide anion, hydroxyl radical and hydrogen peroxide scavenging assay. The brown seaweeds were enzymatically hydrolyzed to prepare water-soluble extracts by using five carbohydrate degrading enzymes (Viscozyme, Celluclast, AMG, Termamyl and Ultraflo) and five proteases (Protamex, Kojizyme, Neutrase, Flavourzyme and Alcalase) of commercial and inexpensive enzymes obtained from Novozyme Co. (Novozyme Nordisk, Bagsvaerd, Denmark). The enzymatic extracts exhibited more prominent effects in hydrogen peroxide scavenging activity (approximately 90%) compared to the other scavenging activities and the activity of enzymatic extracts was even higher than that of the commercial antioxidants. In particular, Ultraflo and Alcalase extracts of S. horneri were dose-dependent and thermally stable. Moreover the two enzymatic extracts strongly inhibited DNA damage (approximately 50%). Those extracts showed significantly (p<0.05) remarkable scavenging effects in DPPH free radical scavenging assay and the activity indicated a marked correlation with phenolic contents. From the results, enzymatic extracts of the brown seaweeds might be valuable antioxidative sources.     

Evaluation of free radical scavenging activity of Butea monosperma Lam

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.     

Free radical and reactive oxygen species scavenging activities of the extracts from seahorse, Hippocampus kuda Bleeler

Seahorse, Hippocampus kuda (SH) a marine teleost fish, is well known not only for its special medicinal composition and used as one of the most famous and expensive materials of traditional Chinese medicine. It was extracted with water (SHW), methanol (SHM), and ethanol (SHE), respectively and evaluated by various antioxidant assays. The including reducing power, total antioxidant, DPPH radical scavenging, hydroxyl radical scavenging, superoxide anion radical scavenging, alkyl radical scavenging, and protective effect on DNA damage caused by hydroxyl radicals generated. Further, the ROS level was detected using a fluorescence probe, 2′,7′-dichlorofluorescin diacetate (DCFH-DA), which could be converted to highly fluorescent dichlorofluorescein (DCF) with the presence of intracellular ROS on mouse macrophages, RAW264.7 cell and inhibited myeloperoxidase (MPO) activity in human myeloid, HL60 cells, respectively. Those various antioxidant activities were compared to standard antioxidants such as α-tocopherol. Among SHM exhibited the highest antioxidant activity in linoleic acid system, effective reducing power, DPPH radical scavenging, hydroxyl radical scavenging, superoxide radical scavenging, alkyl radical scavenging, inhibitory intracellular ROS, and inhibited MPO activity. Furthermore, MTT assay showed no cytotoxicity on mouse macrophages cell (RAW264.7) and human cell lines (MRC-5, HL60, U937). This antioxidant property depends on concentration and increasing with increased amount of extracts. The results obtained in the present study indicated that the see horse (Hippocampus kuda Bleeker) is a potential source of natural antioxidant.     

河南产景天三七抗氧化活性研究

用清除二苯代苦味酰基(DPPH)自由基、2,2′-连氨-(3-乙基苯并噻唑啉-6-磺酸)二氨盐(ABTS)自由基和铁离子还原/抗氧化能力(FRAP)测定法对洛阳和信阳产景天三七(Sedum aizoon)体外总抗氧化活性进行评价,并与阳性对照丁基羟基茴香醚(BHA)、没食子酸丙酯(PG)和二丁基羟基甲苯(BHT)比较。发现洛阳和信阳景天三七有较好的体外抗氧化活性。洛阳和信阳产乙酸乙酯部位活性最强,正丁醇部位活性次之。在6个提取部位中,信阳产景天三七乙酸乙酯部位活性最强,其清除DPPH、ABTS自由基能力强于阳性对照BHA和BHT,还原Fe3+的能力大于阳性对照BHT,洛阳产景天三七乙酸乙酯部位和正丁醇部位活性次之。     

In vitro antioxidant studies in leaves of Annona species

Antioxidant potential of leaves of three different species of Annona was studied by using different in vitro models eg., 1,1-diphenyl-2-picryl hydrazyl (DPPH), 2,2-azinobis-(3-ethylbenzothizoline-6-sulphonate) (ABTS), nitric oxide, superoxide, hydroxy radical and lipid peroxidation. The ethanolic extract of A. muricata at 500 microg/ml showed maximum scavenging activity (90.05%) of ABTS radical cation followed by the scavenging of hydroxyl radical (85.88%) and nitric oxide (72.60%) at the same concentration. However, the extract showed only moderate lipid peroxidation inhibition activity. In contrast, the extract of A. reticulata showed better activity in quenching DPPH (89.37%) and superoxide radical (80.88%) respectively. A.squamosa extract exhibited least inhibition in all in vitro antioxidant models excepting hydroxyl radical (79.79%). These findings suggest that the extracts of A. muricata possess potent in vitro antioxidant activity as compared to leaves of A. squamosa and A. reticulata suggesting its role as an effective free radical scavenger, augmenting its therapeutic     

Evaluation of antioxidant and neuroprotective effect of Hippophae rhamnoides (L.) on oxidative stress induced cytotoxicity in human neural cell line IMR32

Aim and objectiveHippophae rhamnoides is an edible, nutrient rich plant found in the northern regions of India. It belongs to the family Elaeagnaceae and is well known for its traditional pharmacological activities. The present study was aimed to investigate the antioxidant and neuroprotective activities of H. rhamnoides.MethodologyThe hydroalcoholic extract of H. rhamnoides was evaluated for free radical scavenging activity using DPPH, hydroxyl radical scavenging and ferric thiocyanate assays. In vitro neuroprotective activity was assessed on human neuroblastoma cell line-IMR32 against hydrogen peroxide (H₂O₂) induced cytotoxicity. The neuroprotective effect was determined by measuring the cell viability through tetrazolium dye MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) reducing assay and propidium iodide (PI) staining. Also the intracellular reactive oxygen species (ROS) activity was assessed using dichloro-dihydro-fluorescein diacetate (DCFDA) assay by flowcytometer.ResultsThe results of the study demonstrated that H. rhamnoides extract possesses potential free radical scavenging activity. The IC₅₀ value for DPPH and OH radical scavenging assay was 70.92 μg/ml and 0.463 mg/ml, also the extract was also found to have considerable level of lipid peroxidation activity. The neuroprotective effect of H. rhamnoides was confirmed by its cell viability enhancing capacity against hydrogen peroxide induced cell cytotoxicity. The extract acted on IMR32 cells in a dose dependent manner as observed through PI and MTT assays. The percentage intracellular ROS activity was reduced by 60–70% in treated cells compared to H₂O₂ control.ConclusionThus the outcome of the study suggests that H. rhamnoides acts as a neuroprotectant against oxidative stress induced neurodegeneration.     

Inhibitory effect of Teucrium ramosissimum extracts on aflatoxin B1, benzo[a]pyrene, 4-nitro-o-phenylenediamine and sodium azide induced mutagenicity: Correlation with antioxidant activity

The mutagenic potential of total oligomers flavonoids (TOF), ethyl acetate (EA) and petroleum ether (PE) extracts from aerial parts of Teucrium ramosissimum was assessed using Ames Salmonella tester strains TA98, TA100 and TA1535 with and without metabolic activation (S9). None of the different extracts produced a mutagenic effect. Likewise, the antimutagenicity of the same extracts was tested using the “Ames test”. Our results showed that T. ramosissimum extracts possess antimutagenic activity against all the tested genotoxicants (aflatoxin B₁, benzo[a]pyrene, 4-nitro-o-phenylenediamine and sodium azide) in the Salmonella assay systems used in this study. In addition, all extracts showed important free radical scavenging activity toward the radicals DPPH and ABTS except the PE extract.     

荸荠皮提取物对DPPH自由基清除活性

采用70%丙酮溶液对荸荠皮中抗氧化物质进行提取,得到红棕色浸膏。通过定性及定量方法分析了荸荠皮提取物中可能存在的具有抗氧化活性的物质;采用DPPH自由基法测定了荸荠皮提取物对DPPH自由基的清除能力。结果显示,荸荠皮提取物中含有多酚类和黄酮类等化合物,其多酚含量为3.31%(w/w,以干物质计)。DPPH自由基法显示,荸荠皮提取物具有一定的清除DPPH自由基能力,其清除能力与提取物浓度之间显示出良好的剂量-效应关系。该提取物(IC50值为130.37 ppm)对DPPH自由基清除能力略低于BHT(IC50值为94.16 ppm)。     

铁皮石斛化学成分及其抗氧化活性研究

采用硫酸-苯酚法、AlCl^₃比色法、酸性染料比色法测定铁皮石斛(Dendrobium officinale)花、叶、茎中多糖、黄酮、生物碱含量，通过DPPH和ABTS清除实验评价铁皮石斛花、叶、茎的水提物和乙醇提取物的体外抗氧化活性。结果表明，铁皮石斛不同部位的多糖含量茎>花>叶，黄酮含量花>叶>茎，生物碱在各个部位分布均较少。其中茎的多糖含量可达23.92%，花中黄酮含量可达1.847%。抗氧化能力评价表明，铁皮石斛花水提物、茎醇提物、花醇提物的DPPH自由基清除能力相对较好，半效应浓度(EC₅₀)分别为410.4 μg·mL^-1、454.1 μg·mL^-1、573.2 μg·mL^-1；铁皮石斛茎醇提物、花醇提物、花水提物ABTS自由基清除能力相对较好，半效应浓度(EC₅₀)分别为61.1 μg·mL^-1、62.2 μg·mL^-1、103.0 μg·mL^-1。铁皮石斛花的提取物抗氧化活性整体优于叶和茎，醇提物抗氧化能力优于水提物。