• Title/Summary/Keyword: ORAC assay

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Microplate-Based Oxygen Radical Absorbance Capacity (ORAC) Assay of Hydrophilic and Lipophilic Compartments in Plasma

  • Kwak Ho Kyung;Blumberg Jeffrey B.;Chen Chung Yen;Milbury Paul E.
    • Nutritional Sciences
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    • v.9 no.1
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    • pp.48-54
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    • 2006
  • Methods have been developed to evaluate the total antioxidant capacity of foods and plasma but limitations are associated with their ability to determine precisely the contribution of lipophilic antioxidants in a lipid milieu as well as interactions among them Thus, we modified the Oxygen Radical Absorbance Capacity (ORAC) assay to determine the peroxyradical scavenging ability of both hydrophilic and lipophilic compartments in plasma The hydrophilic ORAC assay was performed in a phosphate buffer system utilizing 2,2'-azobis (2-amidinopropane) dihydrochloride as a peroxyradical generator and fluorescein as the target The lipophilic ORAC assay was carried out in a dimethylsulfoxide :butyronitrile (DMSO/BN, 9:1 v/v) system using 2,2'-azobis (2,4-dimethyl valeronitrile) as a peroxyradical generator and BODIPY C11 581/591 as the target Analyses were conducted in bovine serum supplemented with water - and lipid - soluble antioxidants and in human plasma. Albumin (0.5$\sim$5 g/dL) and uric acid (0.1$\sim$0.5 $\mu$mol/L) increased hydrophilic ORAC values in a dose-dependent fashion ($R^{2}$=0.97 and 0.98, respectively) but had no impact on lipophilic ORAC values. $\alpha$-Tocopherol (15$\sim$200 $\mu$mol/L) increased lipophilic ORAC values in a dose-dependent fashion ($R^{2}$=0.94); neither $\alpha$-tocopherol nor $\beta$-carotene had an impact on hydrophilic ORAC values. However, addition of $\beta$-carotene at physiological concentration (0.23$\sim$1.86 $\mu$mol/L), either alone or in combination with other carotenoids, had no significant impact on lipophilic ORAC values. Thus, while assays of 'total antioxidant capacity' in biological matrices would be a useful research and clinical tool, existing methods are limited by the lack of complete responsiveness to the full range of dietary antioxidants.

Determination of the Antioxidant Capacity of Korean Ginseng Using an ORAC Assay (ORAC Assay 에 의한 인삼의 항산화 활성 연구)

  • Kim, Sung-Hwan;Kim, Young-Mok
    • Journal of the East Asian Society of Dietary Life
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    • v.17 no.3
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    • pp.393-401
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    • 2007
  • This study was performed to investigate the antioxidant activity of Korean ginseng using an ORAC(Oxygen Radical Absorbance Capacity) assay. Four fractions each (80% ethanol, ethyl acetate, water saturated 1-butanol, and water) were obtained from different ginseng samples (White Ginseng: ; 6 yrs-., 5 yrs-., ; Cork Ginseng: ; 5 yrs-., 4 yrs-.). The saponin content of each fraction was quantified by LC/MS, and the antioxidant capacity of the ginseng was measured by the ORAC assay. The ORAC method, which was recently validated using automatic liquid handling systems, has been adapted for manual handling with the use of a conventional fluorescence microplate reader. Furthermore, the ORAC assay provides a direct measure of hydrophilic chain-breaking antioxidant capacity against peroxy radical, which is the exiting and emission of 2,2'-Azobis (2-methylpropionamidine)-dihychloride (AAPH). As a result of our experiments, ginsenosides Rg1 and Rb1 were the two major saponins found in the ginseng samples, and Rc, Rb2, Re, Rd, Rg3, and Rh1 were detected in a small quantities. For the antioxidant capacities of the fractions (80% ethanol, ethyl acetate, butanol, and water), we found that the organic solvent fraction had similar antioxidant capacities, and were higher than the capacity of the water fraction. When determining the similarities in each fraction, only the ethyl acetate fraction showed similarity compared to other fractions (p>0.05). The antioxidant capacity of ginseng may come from phenolic compounds and some nonpolar saponins. However, based on the results of this study, we hypothesize that some acidic polysaccharides and other biological components may contribute to its antioxidant capacity. Additional research is required to determine other possible biological response modifiers that contribute to the antioxidant capacity of ginseng.

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Phenolic Compounds from the Twigs of Stewartia pseudocamellia (노각나무 가지의 Phenol성 성분)

  • Bae, Jong Jin;Kwak, Jong Hwan
    • Korean Journal of Pharmacognosy
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    • v.46 no.4
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    • pp.303-308
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    • 2015
  • Ten phenolic compounds were isolated from the twigs of Stewartia pseudocamellia. The isolated compounds were identified as 5,7,3',5'-tetrahydroxyflavanone (1), 3,5,7,3',5'-pentahydroxyflavanone (2), quercetin (3), (+)-aromadendrin (4), (+)-ampelopsin (5), myricetin (6), (+)-catechin (7), (-)-epicatechin (8), kaempferol (9), and fraxin (10) by spectroscopic analysis. Compounds 1, 2, 4, 6, 8, and 9 were isolated from this plant for the first time. The antioxidant activities of compounds 1-10 were evaluated by the DPPH and/or ORAC (oxygen radical absorbance capacity) assay. Compounds 3, 5-9 showed significant antioxidative effects on DPPH assay. Among the active compounds, 6 exhibited higher activity compared to trolox on ORAC assay.

Flavonoids from the Flower of Clerodendrum trichotomum (누리장나무 꽃의 Flavonoid 성분)

  • Lee, Jong-Wook;Kang, Se Chan;Bae, Jong Jin;Lee, Kyung Bok;Kwak, Jong Hwan
    • Korean Journal of Pharmacognosy
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    • v.46 no.4
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    • pp.289-294
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    • 2015
  • Seven flavonoids were isolated from the flower of Clerodendrum trichotomum. Their structures were identified as apigenin (1), genistein (2), chrysoeriol (3), genistein 7-O-glucoside (4), kaempferol 3-O-glucoside (5), isorhamnetin 3-O-glucoside (6) and apigenin 7-O-glucoside (7) on the basis of spectral data. These compounds were isolated from C. trichotomum for the first time. The antioxidant activity of compounds 1-7 were evaluated by the ORAC (oxygen radical absorbance capacity) assay, and the ORAC values were expressed as relative trolox equivalent. All isolated compounds exhibited antioxidant activity.

Evaluation of Anti-Oxidant from Natural Products (천연물로부터 유래한 천연 항산화제 규명)

  • Kwon, Jin-A;Yang, Yoon-Jung;Park, Jong-Hyuk;Kang, Se-Chan
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2011.10a
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    • pp.20-20
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    • 2011
  • In this study, we analyzed 80%MeOH extract of fruits of sorbaria sorbifolia var. stellipila MAX. to measure the total antioxidant capacity by oxygen radical absorbance capacity (ORAC) assay, individual flavonoid content by high-performance liquid chromatography (HPLC). n-Hexane ($1.02{\pm}0.036$), $CH_2Cl_2$ ($0.95{\pm}0.025$), EtOAc ($1.94{\pm}0.065$), n-BuOH ($1.98{\pm}0.054$), D.W. ($1.2{\pm}0.032$) fractions were examined antioxidative activity by ORAC assay. It was revealed that EtOAc($1.94{\pm}0.065$), n-BuOH($1.98{\pm}0.054$) fractions had significant antioxidative activity. The isolation and separation were facilitated using open column chromatography, while separation, purification and identification were accomplished by using high-performance liquid chromatography (HPLC) and nuclear magnetic resonance (NMR).

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Antioxidant Effects and Nitrite Scavenging Ability of Extract from Acanthopanax cortex Shoot (오가피순 추출물의 항산화 효과 및 아질산염 소거능)

  • Yu, Seok-Yeong;Lee, Young-Jun;Song, Ho-Seong;Hong, Hee-Do;Lim, Jeong-Ho;Choi, Hyeon-Son;Lee, Boo-Yong;Kang, Suk-Nam;Lee, Ok-Hwan
    • The Korean Journal of Food And Nutrition
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    • v.25 no.4
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    • pp.793-799
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    • 2012
  • This study was conducted to examine the antioxidative effect and nitrite scavenging ability of extract from Acanthopanax cortex shoot. The total phenolic compound and flavonoids contents of extract from Acanthopanax corex shoot were $116.33{\pm}6.09mg\;GAE/g$ and $65.07{\pm}4.10mg\;RE/g$, respectively. Antioxidative activities were measured by various in vitro models such as DPPH radical scavenging activity, FRAP, reducing power, ABTS radical scavenging activity, ORAC assay. This results showed that the extract of Acanthopanax cortex shoot was effective in scavenging radicals and protecting oxidation when assessed various in vitro systems. Similarly, the nitrite scavenging ability of extract was increased in a dose-dependent manner. Moreover, ORAC value at a concentration of $0.1mg/m{\ell}$ was $103.4{\pm}5.6{\mu}M\;TE/g$. Considering high consumer demand beneficial health effects, Acanthopanax cortex shoot can be utilized to develop functional food health-promoting and natural antioxidant agents.

Antioxidative Constituents from Fruit of Sorbaria sorbifolia var. stellipila MAX. (쉬땅나무(Sorbaria sorbifolia var. stellipila MAX.) 열매의 항산화 활성 성분)

  • Park, Jong-Hyuk;Kwon, Jin-A;Yang, Yoon-Jung;Han, Hyo-Sang;Han, Min-Woo;Lee, Young-Il;Kim, In-Su;Lee, Jong-Ill;Kang, Se-Chan
    • Korean Journal of Plant Resources
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    • v.24 no.4
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    • pp.337-342
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    • 2011
  • The purpose of this study was to evaluated the antioxidative constituents and their activities of the 80% methanolic extracts from fruit of Sorbaria sorbifolia var. stellipila MAX. The isolation of active compound was performed in three steps: solvent partition, open column chromatography, and high-performance liquid chromatography (HPLC). The solvent fractions were tested for their antioxidant activities by oxygen radical absorbance capacity (ORAC). The antioxidant activity of 80% methanolic extracts by various solvent partitions was in the order of 80% MeOH (1.68 ${\pm}$ 0.027), n-hexane (1.02 ${\pm}$ 0.036), $CH_2Cl_2$ (0.95 ${\pm}$ 0.025), EtOAc (1.98 ${\pm}$ 0.065), n-BuOH (1.94 ${\pm}$ 0.054) and Water (1.28 ${\pm}$ 0.032). Therefore, the results indicated that the potential antioxidant activities and functional values were observed significantly at EtOAc fraction from fruit of S. sorbifolia, flavonoid compound isolated.

Comparison of antioxidant activity and prevention of lymphocyte DNA damage by fruit and vegetable juices marketed in Korea (시판 천연 과일주스와 채소주스의 항산화능과 임파구 DNA 손상 방지 효능 비교)

  • Cho, Miran;Lee, Hye-Jin;Kang, Myung-Hee;Min, Hyesun
    • Journal of Nutrition and Health
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    • v.50 no.1
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    • pp.1-9
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    • 2017
  • Purpose: Fruit and vegetable juices are known to be rich sources of antioxidants, which have beneficial effects on diseases caused by oxidative stress. The purpose of this study was to directly compare the antioxidant activities of fruit and vegetable juices marketed in Korea. Methods: We analyzed four fruit juices, two vegetable juices, two yellow-green juices, and six mixed vegetable juices. Antioxidant activities were analyzed using 2,2-diphenyl-1-picrylhydrazyl (DPPH) test, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonate) (ABTS) test, and oxygen radical absorbance capacity (ORAC) assay. Protective effects against DNA damage were determined using an ex vivo comet assay with human lymphocytes. Results: DPPH radical scavenging activities were in the following order: blueberry juice > mixed vegetable C juice > kale juice > mixed vegetable P juice > grape juice. ABTS radical scavenging activities were in the following order: blueberry juice > mixed vegetable C juice > grape juice > mixed vegetable P juice > kale juice. Peroxyl radical scavenging activities as assessed by ORAC assay were in the following order: blueberry juice > kale juice > mixed vegetable C juice > grape juice. Grape or blueberry juice showed strong abilities to prevent DNA damage in lymphocytes, and the difference between them was not significant according to the GSTM1/GSTT1 genotype. Conclusion: Antioxidant activities of fruit and vegetable juices and ex vivo DNA protective activity increased in the order of blueberry juice, grape juice, and kale juice, although the rankings were slightly different. Therefore, these juices rich in polyphenols and flavonoids deserve more attention for their high antioxidant capacity.

Antioxidative Constituents from the Whole Plants of Euphorbia supina (애기땅빈대의 항산화 활성 성분)

  • Hong, Hyun-Kyung;Kwak, Jong-Hwan;Kang, Se-Chan;Lee, Jong-Wook;Park, Jong-Hyuk;Ahn, Jong-Woong;Kang, Hye-Sook;Choung, Eui-Su;Zee, Ok-Pyo
    • Korean Journal of Pharmacognosy
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    • v.39 no.3
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    • pp.260-264
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    • 2008
  • Eight compounds were isolated from the EtOAc soluble fraction of Euphorbia supina MeOH extract as the radical scavengers for antioxidant activity. Their structures were identified as kaempferol (1), quercetin (2), juglanin (3), avicularin (4), astragalin (5), isoquercitrin (6), hyperin (7), and nicotiflorin (8) by spectroscopic analysis. The antioxidant activity was evaluated by the ORAC (oxygen radical absorbance capacity) assay, which measures scavenging activity against peroxy radicals induced by 2,2'-azobis (2-methylpropionamidine) dihydrochloride, and the ORAC value is expressed as relative trolox equivalent. Compounds 4, 6, and 7 exhibited potent antioxidant activity, whereas the other compounds showed weaker activity than trolox.

Comparative Study on Biological Activities of Colored Potatoes, Hongyoung and Jayoung Cultivar (유색감자 홍영 및 자영 추출물의 생물학적 활성 비교)

  • Kang, Se-Chan; Choung, Myoung-Gun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.53 no.2
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    • pp.233-238
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    • 2008
  • This experiment was conducted to enhance the colored potatoes utilization and to determine the biological activity of colored potato extracts. In order to understand the factors responsible for the potent anti-oxidant ability of colored potatoes, it has been evaluated for anti-oxidative activity using oxygen radical absorbance capacity (ORAC) assay. 'Hongyoung' extract was significant anti-oxidant activities in ORAC assay. About two-fold higher radical absorbance capacity was found in 'Hongyoung' compared to that in 'Jayoung'. The ability of 80% ethanol extracts from colored potatoes to influence the inhibitory activity of nitric oxide (NO) and nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) has also been investigated. The various therapeutic benefit claims in the new functional medicinal usage of colored potatoes ascribed to the phenolic compounds and anthocyanin. This result revealed that the extracts of colored potatoes are expected to be good candidate for development into sources of free radical scavenger or COX-2 inhibiting agents.