• 제목/요약/키워드: oxygen radical absorbance capacity(ORAC) assay

검색결과 45건 처리시간 0.025초

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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    • 제9권1호
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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.

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

  • 김성환;김영목
    • 동아시아식생활학회지
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    • 제17권3호
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    • pp.393-401
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    • 2007
  • 인삼의 여러 생리 활성 가운데 항산화 정도를 알아보기 위하여 백삼(6 년근), 백삼(5 년근), 피부백삼(5 년근), 피부백삼 (4 년근) 의 80% 에탄올 엑기스, 에틸아세테이트 분획, 수포화 부탄올 분획, 물 분획을 얻은 후 LC/Mass 를 사용하여 사포닌 함량을 조사하고 기존의 여러 가지 항산화 작용 측정 방법들의 오류를 없애고 더욱 정확한 결과를 낼 수 있는 대처 방안으로 선정된 ORAC Assay에 의해 항산화 활성을 검토하였다. 인삼 중 사포닌은 ginsenoside Rg 1 과 Rb1 이 주요 성분으로 다량 함유하고 있었으며, Rc, Rb2, Re 등이 뒤를 이었고, 그밖에도 Rd, Rg3, Rh1가 널리 분포하고 있었다. 피부백삼 5 년근의 경우 에탄올 엑기스와 수포화 부탄올 분획에서 다른 인삼 분획에 비해 높은 함유량을 보였으나 실험의 한계상 인삼 재배기간과 인삼 종류별 각각의 분획에 대한 사포닌 함량 비교는 어려웠다. 인삼의 각 분획별 항산화 활성은 80% 에탄올 엑기스, 에틸아세테이트 분획, 수포화 부탄올 분획, 물 분획 모두에서 나타났고 비교적 전체 유기 용매 분획의 값이 비슷하였으며, 수층 분획이 다소 낮은 활성을 보였다. 검체 인삼들의 각 용매추출 분획 상호간의 유의성 비교에서는 모든 인삼 검체의 에틸아세테이트 분획에서만 유의성을 나타내었다(p>0.05). 인삼 중 항산화 활성은 에틸아세테이트 층으로 이행되는 폴리페놀 계통 성분이나 일부 비극성의 사포닌에 의한 것으로 추측되고 있으나 모든 분획에서 나타난 것으로 보아 이들 외에 산성 다당체, 당 단백질, 수용성 다당류 등 다른 생리활성 물질에 대한 연구가 요구된다.

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In vitro and Cellular Antioxidant Activity of Arginyl-fructose and Arginyl-fructosyl-glucose

  • Lee, Jung-Sook;Kim, Gyo-Nam;Lee, Sang-Hyun;Kim, Eui-Su;Ha, Kyoung-Soo;Kwon, Young-In;Jeong, Heon-Sang;Jang, Hae-Dong
    • Food Science and Biotechnology
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    • 제18권6호
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    • pp.1505-1510
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    • 2009
  • Arginyl-fructose (AF) and arginyl-fructosyl-glucose (AFG) were chemically synthesized and purified. Their in vitro and cellular antioxidant activity was investigated using oxygen radical absorbance capacity (ORAC) and cellular antioxidant activity assay, respectively. The peroxyl radical scavenging activity of AF was much higher than that of AFG, which was in good agreement with their reduction capacity to donate electrons or hydrogen atoms. On the other hand, the hydroxyl radical scavenging activity of AF was weaker than that of AFG, which was consistent with their metal chelating activity, suggesting that AFG-$Cu^{2+}$ complex may be less redox-active than AF-$Cu^{2+}$ complex due to 1 glucose molecule attached. The cellular antioxidant activity of AF and AFG appeared to depend on both their permeability into cell membrane and the scavenging activity on peroxyl or hydroxyl radicals. These results indicate that AF and AFG, Maillard reaction products, may have a high potential as a material for the development of nutraceutical food with antioxidant activity.

Inhibition of Matrix Metalloproteinases-12 (MMP-12) and Anti-oxidant Effect of Xanthohumol from Hop (Humulus lupulus L.)

  • Lee, Keyong Ho;Yoon, Won Ho
    • Natural Product Sciences
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    • 제18권4호
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    • pp.261-265
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    • 2012
  • Xanthohumol was isolated from hops (Humulus lupulus L.), and then investigated anti-oxidant effect by AAPH-induced LLC-PK1 cell and oxygen radical absorbance capacity (ORCA) assays and MMP-12 inhibitory effect by direct MMP-12 inhibition assay. The treatment of xanthohumol protected LLC-PK1 cells from AAPH-induced cell damage such as cell viability, SOD and GSH-px reduction in a dose dependant manner (0.1, 1, and $5{\mu}M$), the SOD value was 2.98, 4.51, and 5.77 U/mg protein, and GSH-px value was 30.12, 49.32, and 60.11 U/mg protein. ORAC value of xanthohumol was showed as 4320, 12004, and $14209{\mu}M$ TE/g at the concentration 0.1, 1, and $5{\mu}M$, respectively. The change of SOD and GSH-px values was significantly correlated with the results of ORAC assay, that is, AAPH-induced cell and ORCA assays. In addition, inhibition of MMP-12 that is known to play an important role in skin aging was 14%, 37%, 46%, and 79% at the concentration of 0.01, 0.1, 1, and $5{\mu}M$, respectively. On the basis of these results, xanthohumol from hops (Humulus lupulus L.) showed interesting biological and pharmacological activity such as anti-oxidant effect and anti-aging.

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

  • 배종진;곽종환
    • 생약학회지
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    • 제46권4호
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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.

Evaluation of the Antioxidant Activities of Natural Components of Artemisia iwayomogi

  • Yan, Xi-Tao;Ding, Yan;Lee, Sang Hyun;Li, Wei;Sun, Ya-Nan;Yang, Seo Young;Jang, Hae Dong;Kim, Young Ho
    • Natural Product Sciences
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    • 제20권3호
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    • pp.176-181
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    • 2014
  • The antioxidant activities of 29 components isolated from the aerial parts of Artemisia iwayomogi were evaluated in vitro and in cell culture. Among the tested compounds, 2, 6, 8, 10, 13, and 14 exhibited the greatest peroxyl radical-scavenging activities in the oxygen radical absorbance capacity (ORAC) assay, and 2, 10, and 14 also showed significant reducing capacities. However, all compounds showed weak metal chelating activities. Their cellular antioxidant activities were evaluated in HepG2 cells. At $10{\mu}M$, compounds 6, 8, and 14 exhibited stronger protection against 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative stress than compounds 2, 10, and 13. Moreover, Compounds 2 and 8 were more effective in protecting against $Cu^{2+}$-induced oxidative stress than compounds 6, 10, 13, and 14 at $10{\mu}M$. These results suggest that the phenolic compounds in A. iwayomogi have the potential to be developed as natural antioxidants for the treatment of oxidative stress-related diseases.

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

  • 이종욱;강세찬;배종진;이경복;곽종환
    • 생약학회지
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    • 제46권4호
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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.

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

  • 권진아;양윤정;박종혁;강세찬
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2011년도 임시총회 및 추계학술발표회
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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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접골목 추출물에 의한 항산화 활성이 정상 간세포의 t-BHP 유발 산화스트레스에 미치는 영향 (Antioxidant Activity and Protective Effects of Extracts from Sambucus williamsii var. coreana on t-BHP Induced Oxidative Stress in Chang cells)

  • 김기태
    • 대한한의진단학회지
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    • 제17권3호
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    • pp.275-286
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    • 2013
  • In the present study, antioxidant activity and protective effect of extracts from Sambucus williamsii var. coreana stems (SWC) were evaluated on tert-butyl hydroperoxide (t-BHP) induced oxidative stress in human liver (Chang) cells. Antioxidant activities of the SWC extracts were determined by various radical scavenging activities, such as DPPH, ferric reducing antioxidant power (FRAP), 2,2'-azinobis-(3-ethybenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity and oxygen radical absorbance capacity (ORAC) assay. SWC extracts showed strong antioxidant effect on various assay. To determine the hepatoprotective effects of SWC on t-BHP induced oxidative damage, cell viability was measured using MTT assay. Pretreatment of SWC extracts showed increasing cell viability, decreasing ROS and restoring mitochondria membrane potential on t-BHP induced oxidative stress in Chang cells. Our findings suggest that SWC extracts may be considered a potential agent for therapeutic protective effect from oxidative stress through its antioxidant activity.

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

  • 홍현경;곽종환;강세찬;이종욱;박종혁;안종웅;강혜숙;정의수;지옥표
    • 생약학회지
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    • 제39권3호
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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.