• Title/Summary/Keyword: DPPH assay

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Antioxidant Activity and Protection from DNA Damage by Water Extract from Pine (Pinus densiflora) Bark

  • Jiang, Yunyao;Han, Woong;Shen, Ting;Wang, Myeong-Hyeon
    • Preventive Nutrition and Food Science
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    • v.17 no.2
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    • pp.116-121
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    • 2012
  • Water extract from Pinus densiflora (WPD) was investigated for its antioxidant activity and its ability to provide protection from DNA damage. A series of antioxidant assays, including a 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical-scavenging assay, a reducing power assay, a metal-chelating assay, a superoxide radical scavenging assay, and a nitrite scavenging ability, as well as a DNA damage protection assay were performed. Total phenolic content was found to be 211.32 mg Tan/g WPD. The extract scavenged 50% DPPH free radical at a concentration of 21.35 ${\mu}g/mL$. At that same concentration, the reducing power ability of WPD was higher than that of ${\alpha}$-tocopherol. The extract chelated 68.9% ferrous ion at the concentration of 4 mg/mL. WPD showed better nitrite scavenging effect at the lower pH. Meanwhile, WPD exhibited a strong capability for DNA damage protection at 1 mg/mL concentration. Taken together, these data suggest water extract from Pinus densiflora could be used as a suitable natural antioxidant.

Effect of Arctigenin on Tyrosinase Activity and Melanin Production in B16 Melanoma Cells (B16 Melanoma 세포에서 Arctigenin이 Tyrosinase 활성과 Melanin 생성에 미치는 영향)

  • Lee, Dong Ja;Sim, Sang Soo
    • YAKHAK HOEJI
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    • v.56 no.6
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    • pp.395-400
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    • 2012
  • To investigate the possibility of development as a whitening agent using arctigenin, we measured DPPH assay, NBT/XO assay, intracellular ROS scavenging assay, tyrosinase assay and MSH-induced melanin production in B16 melanoma cells. Arctigenin dose-dependently had anti-oxidant activity in DPPH, NBT/XO and intracellular ROS assay. Although arctigenin did not inhibit purified tyrosinase activity, it dose-dependently inhibited tyrosinase activity and melanin production in B16 melanoma cells stimulated by $1{\mu}M$ ${\alpha}$-MSH. In particular, arctigenin at a concentration $100{\mu}M$ inhibited ${\alpha}$-MSH-stimulated tyrosinase activity and melanin production by $50.9{\pm}2.9%$ and $69.0{\pm}6.5%$ respectively. And typical tyrosinase inhibitor, arbutin, inhibited $57.7{\pm}2.9%$ and $65.1{\pm}5.0%$ respectively. Such an similar inhibitory effect of arctigenin and arbutin in B16 melanoma cells may be due to the inhibition of MSH signal pathway rather than the direct inhibition of tyrosinase. Therefore, these results suggest that arctigenin may be useful for the development as whitening agents.

Comparison of lymphocyte DNA damage levels and total antioxidant capacity in Korean and American diet

  • Lee, Min Young;Kim, Hyun A;Kang, Myung Hee
    • Nutrition Research and Practice
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    • v.11 no.1
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    • pp.33-42
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    • 2017
  • BACKGROUND/OBJECTIVE: This study aims to measure the in vitro antioxidant capacity of Korean diet (KD) with American diet (AD) as a control group and to examine the ex vivo DNA damage reduction effect on human lymphocytes. MATERIALS/METHODS: The KD applied in this study is the standard one-week meals for Koreans (2,000 kcal/day) suggested by 2010 Dietary Reference Intakes for Koreans. The AD, which is the control group, is a one-week menu (2,000 kcal/day) that consists of foods that Americans would commonly take in according to the National Health and Nutrition Examination Survey. The antioxidant capacity of each menu was measured by means of the total phenolic assay and 3 in vitro antioxidant activity assays (2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, trolox equivalent antioxidant capacity (TEAC), Oxygen radical absorbance capacity ($ORAC_{ROO{\cdot}}$)), while the extent of ex vivo lymphocyte DNA damage was measured by means of the comet assay. RESULTS: When measured by means of TEAC assay, the in vitro antioxidant capacity of the KD of the day was higher than that of the AD (P < 0.05) while there was no significant difference in total phenolic contents and DPPH and ORAC assays. The ex vivo lymphocyte DNA damage protective effect of the KD was significantly higher than that of the AD (P < 0.01). As for the one-week menu combining the menus for 7 days, the total phenolic assay (P < 0.05) and in vitro antioxidant capacity (P < 0.001, DPPH; P < 0.01, TEAC) of the KD menu were significantly higher than those of the AD menu. Likewise, the ex vivo DNA damage reduction rate of the Korean seven-day menu was significantly higher than that of the American menu (P < 0.01). CONCLUSION: This study demonstrates that the high antioxidant capacity and DNA damage protective effect of KD, which consists generally of various plant foods, are higher than those of typical AD.

Antioxidant and Anti-wrinkling Effects of Extracts from Nelumbo nucifera leaves (하엽(荷葉) 추출물이 항산화 효능 및 피부노화에 미치는 영향)

  • Park, Chan-Ik;Park, Geun-Hye
    • The Korea Journal of Herbology
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    • v.31 no.4
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    • pp.53-60
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    • 2016
  • Objective : The purpose of this study was to investigate anti-aging and antioxidant effects of extracts of Nelumbo nucifera leaves (NN-L) using ethanol on skin .Methods : Each part of leaves(NN-L), flowers(NN-F) and stem(NN-S) was extracted with 70% ethanol. We performed radical scavenging assay(DPPH, ABTS+, Superoxide anion radical), elastase inhibition assay, collagenase inhibition assay. NN-L extracts were tested for cell viability(MTT assay), MMP-1 inhibition and MMP-1 protein expression on CCD-986sk cells (human fibroblast line).Results : Recently, many studies have reported that elastin is also involved in inhibiting or repairing wrinkle formation, although collagen is a major factor in the skin wrinkle formation. We measured its free radical scavenging activity, elastase inhibitory activity and expression of MMP-1 (matrix metalloprotease-1) in human fibroblast cells. Among the parts of Nelumbo nucifera, NN-L showed the highest antioxidant activities and in radical scavenging. DPPH, ABTS+ and Superoxide anion radical scavenging activity of NN-L at concentration of 1,000 μg/mL were 91.43%, 99.31% and 73.7% respectively. In vitro elastase and collagenase inhibition effects of NN-L at concentration of 1,000 μg/mL was 42.8% and 55.3% respectively. The ethanol extract of NN-L showed cell viability of 95.4% in 50 μg/mL concentration. In addition, The results from Western blot assay showed that NN-L decreased the expression of MMP-1 protein in a dose-dependent manner (by up to 35.0% at 50 μM).Conclusion : The findings suggest that the NN-L great potential as a cosmeceutical ingredient with antioxidant and anti-wrinkling effects.

Antioxidant Activity of Orange Flesh and Peel Extracted with Various Solvents

  • Park, Jae-Hee;Lee, Minhee;Park, Eunju
    • Preventive Nutrition and Food Science
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    • v.19 no.4
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    • pp.291-298
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    • 2014
  • The aim of this study was to investigate the antioxidant activity of orange (Citrus auranthium) flesh (OF) and peel (OP) extracted with acetone, ethanol, and methanol. Antioxidant potential was examined by measuring total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity (RSA), total radical-trapping antioxidant potential (TRAP), oxygen radical absorbance capacity (ORAC), and cellular antioxidant activity (CAA). The comet assay was used to determine the protective effects of OF and OP against $H_2O_2$-induced DNA damage. TPC was highest in the acetone extracts of OF and OP. DPPH RSA was also higher in the acetone extracts than in the ethanol extracts. The DPPH RSA was highest in the acetone extracts of OF. The TRAP and ORAC values of the all extracts increased in a dose-dependent manner. In the TRAP assay, the acetone extracts of OF and OP had the lowest $IC_{50}$ values. In the CAA assay, the methanol and acetone extracts of OP had the lowest $IC_{50}$ values. All of the samples protected against $H_2O_2$-induced DNA damage in human leukocytes, as measured by the comet assay, but the acetone extracts of OP had the strongest effect. These results suggest that acetone is the best solvent for the extraction of antioxidant compounds from OF and OP. Furthermore, the high antioxidant activity of OP, which is a by-product of orange processing, suggests that it can be used in nutraceutical and functional foods.

Study on DPPH Free Radical Scavenging and Lipid Peroxidation Inhibitory Activities of Vietnamese Medicinal Plants

  • Phan, Thi Anh Dao;Nguyen, Xuan Hai;Nguyen, Trung Nhan;Tran, Le Quan;Nguyen, Thi Thanh Mai
    • Natural Product Sciences
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    • v.18 no.1
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    • pp.1-7
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    • 2012
  • Among 90 Vietnamese medicinal plant extracts investigated for their antioxidant activity by DPPH assay at various concentrations from $10-100{\mu}g/mL$, 67 showed an inhibition rate over 50% at $100{\mu}g/mL$; 47 had greater than 50% inhibition at $50{\mu}g/mL$; 17 showed over 50% inhibition at $25{\mu}g/mL$. 8 extracts which exhibited strong inhibitory activity more than 50% inhibition at $10{\mu}g/mL$ were further tested for lipid peroxidation inhibition by TBA assay. They displayed activity with $IC_{50}$ values from 30.6 to $158.9{\mu}g/mL$. Until now, this is the first report on antioxidant activity of the female flower of Borassus flabellifer, and the stem of Combretum latifolium, Embelia ribes, Spatholobus parviflorus, and Tetrastigma erubescens. Fractionations of the EtOAc extract prepared from S. parviflorus led to the isolation of protocatechuic acid (1), ferulic acid (2), epicatechin (3), and gallic acid (4). These compounds showed significant DPPH inhibitory activity with $IC_{50}$ values from 6.5 to $23.6{\mu}M$.

Comparison of in vitro antioxidant capacities of Phragmites communis Trin. and Phragmites japonica Steud.

  • Kyeong Won Yun;Kyoung Sun Seo
    • Food Science and Preservation
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    • v.30 no.6
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    • pp.960-968
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    • 2023
  • The rhizome of Phragmites communis Trin. is used for vomiting and belching by clearing stomach and the sprout is used as tea. Phragmites japonica is similar with P. communis except the color of sheath is purple. This study is aimed to compare the in vitro antioxidant activity, total polyphenol and flavonoids contents of P. communis and P. japonica. The antioxidant activities of fractions from the two Phragmites plants were evaluated by 1,1-diphenyl-2- picrylhydrazyl (DPPH) and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging assay. The antioxidant activity varied with plant parts and extract solvents. The fractions of leaf extract from the two Phragmites plants (4.06±1.32-16.47±1.28%) showed higher antioxidant activity by DPPH assay compared with rhizome fractions of two Phragmites plants (0.00±0.00-14.15±0.07%), these are lower compared with ascorbic acid and butylhydroxyanisole (BHA). The highest ABTS radical scavenging activity was found for rhizome ether fraction, namely 74.95±0.56% and 73.04±1.85% for P. communis and P. japonica, these are higher than BHA. The total polyphenol and flavonoids contents were different with plant parts and extract solvents, likewise antioxidant activity. A significant correlation was shown between DPPH and ABTS radical scavenging activity. Considering the results of this study, the leaves and stems of P. communis and P. japonica are expected to be used as natural antioxidants.

Separation and Purification of Effective Components from the Alisma orientale and its Application as a Cosmeceutical Ingredient (택사추출물의 성분분리와 화장품 원료로서의 특성)

  • Lee, Dae-Woo;Kim, Young-Jin;Kim, Young-Sil;Kim, Jong-Heon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.32 no.1 s.55
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    • pp.23-28
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    • 2006
  • In this study, we performed anti-oxidation, whitening, cell recovery and anti-inflammation effects with Alisma orientale to evaluate the cosmeceutical properties. Alisma orientate extract (30, 70, 100 % MeOH) exhibited a significant tree radical scavenging effect against 1,1-diphenyl-2-picryl hydrazine (DPPH) radical generation and showed tyrosinase inhibition effect in a dose dependent manner (over 0.5% concentration). In cell proliferation assay using human fibroblast, it didn't show any proliferation effect but showed safety from cytotoxicity under 0.05% concentration. For whitening assay, we evaluated the melanin synthesis rate using B16 melanocyte and it showed a significant inhibitory effect (up to 40% under 0.05% concentration). After major screening assay, we separate 3 fractions from Alisma orientate extract by MPLC and performed cell recovery assay, melanin synthesis inhibition assay and anti-inflammatory assay. The third fraction showed a cell recovery effect over 30% against radical damage and remarkable repression in melanin synthesis and COX-2 synthesis.

The Anti-inflammatory Effect and DPPH Free Radical Scavenging Capability of Rhizoma drynariae Aqueous Extract (골쇄보 열수추출물 약침액의 항염증반응 및 DPPH 라디칼 소거능 활성에 대한 연구)

  • Kim, Yong-Min;Kim, Ee-Hwa
    • Korean Journal of Acupuncture
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    • v.32 no.4
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    • pp.169-176
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    • 2015
  • Objectives : This study is to investigate the effects of Rhizoma drynariae aqueous extract(RDA) on cell cytotoxicity, Nitric Oxide (NO) and Prostaglandin $E_2(PGE_2)$ production and 1,1-diphenyl-2-picryl ghdrazyl(DPPH) free radical scavenging capability. Methods : Cell cytotoxicity was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay. The production of NO was measured by Griess assay. The production of $PGE_2$ was measured by immunoassay. And, the anti-oxidant activity was measured by the DPPH method. Results : Cell cytotoxicity in 50, 100, 200 and $400{\mu}g/ml$ RDA did not increase significantly compared to the RDA untreated group. RDA($200{\mu}g/ml$ and $400{\mu}g/ml$) inhibited NO and $PGE_2$ production in lipopolysaccharide-stimulated RAW 264.7 cells. RDA had high DPPH free radical scavenging capability. Conclusions : This study indicates that RDA inhibits NO and $PGE_2$ production in lipopolysaccharide-stimulated RAW 264.7 cells and improve DPPH free radical scavenging capability. RDA may have an anti-inflammation effect and an anti-oxidant activity.

Comparison of Antioxidant and Antimicrobial Activities in Siraegi (Dried Radish Greens) according to Cooking Process (조리 과정 중 시래기의 항산화 활성 및 항균 활성 비교)

  • Park, Cho-Hee;Kim, Kyoung-Hee;Yook, Hong-Sun
    • The Korean Journal of Food And Nutrition
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    • v.27 no.4
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    • pp.609-618
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    • 2014
  • We evaluated the yield of extract, antioxidant compounds (total phenolic, total flavonoid), antioxidant (DPPH assay, ABTS assay, Oyaizu's assay, FRAP assay), and antimicrobial activities of Siraegi (dried radish greens) according to cooking process (non-blanched, blanched, seasoned). The yield of non-blanched Siraegi was 4.91%, blanched Siraegi was 0.33%, and seasoned Siraegi was 7.55%. In total polyphenol and flavonoid contents, seasoned Siraegi extracts showed higher antioxidant compounds ($129.85{\pm}0.62mg$ GAE/100 g FW, $35.56{\pm}1.19mg$ CHE/100 g FW) than non-blanched and blanched. Total antioxidant activities (DPPH assay, ABTS assay, FRAP assay, reducing power) were shown to be in the order of seasoned Siraegi > non-blanched Siraegi > blanched Siraegi. In antimicrobial activity, non-blanched Siraegi (5, 10 mg/disc) showed antimicrobial activity against B. cereus, E. cloacae, and E. coli (9.25 mm), and P. aeruginosa (9 mm) at 10 mg/disc. In terms of antimicrobial activity, non-blanched Siraegi was good but eating the dried vegetable was difficult so it is essential to blanch it. Also, with many added seasonings in the process of cooking, it can be easy to eat. Overall, the results of this study demonstrate that cooked Siraegi with seasoning would be the most efficient way of ingesting the antioxidant material.