• Title/Summary/Keyword: ORAC assay

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Comparison of Antioxidant Activity and ${\alpha}$-Glucosidase Inhibiting Activity by Extracts of Galla rhois

  • Lee, Seung-Hyun;Lee, Sang-Han
    • Current Research on Agriculture and Life Sciences
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    • v.31 no.4
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    • pp.227-232
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    • 2013
  • We studied antioxidant activity and inhibitory effect of ${\alpha}$-glucosidase from aqueous, ethanolic and methanolic fractions of Galla rhois. In FRAP and ORAC assay for measuring antioxidant activity, we confirmed that Galla rhois extracts had strong antioxidant activity and ethanolic and methanolic extracts were relatively stronger than aqueous extract. We used trolox as a positive control. In order to measure the inhibitory effect of ${\alpha}$-glucosidase, we compared acarbose and Galla rhois extracts. As a result of ${\alpha}$-glucosidase inhibitory assay, aqueous, ethanolic and methanolic extracts of Galla rhois showed high inhibitory activitity and ethanolic and methanolic extracts were relatively stronger than aqueous extract. The 50% inhibitory concentrations (IC50s) of acarbose, aqueous, ethanolic and methanolic fractions were 0.45 mM, $0.53{\mu}g/ml$, $0.415{\mu}g/ml$ and $0.37{\mu}g/ml$, respectively. These results suggest that Galla rhois extracts can be a clinically useful anti-diabetic ingredient, indicating that it needs to be fractionated and isolated and should be further investigated.

Validation of Analytical Methods for Plasma Total Antioxidant Capacity by Comparing with Urinary 8-Isoprostane Level

  • Lee, Sang Gil;Wang, Taoran;Vance, Terrence M.;Hurbert, Patrice;Kim, Dae-Ok;Koo, Sung I.;Chun, Ock K.
    • Journal of Microbiology and Biotechnology
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    • v.27 no.2
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    • pp.388-394
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    • 2017
  • Although several analytical methods for measuring total antioxidant capacity (TAC) have been applied to biological samples, there were often dissimilar results due to the different principles of methods applied. Thus, this study aimed to validate four conventional analytical methods for measuring plasma TAC, including the ABTS assay, DPPH assay, FRAP assay, and ORAC assay, by comparing with urinary 8-isoprostane concentration. In addition, TAC results were compared with antioxidant enzyme activities including superoxide dismutase (SOD) and glutathione peroxidase in erythrocyte, and catalase in plasma. Plasma TAC measure by ABTS assay was strongly correlated with the result by FRAP assay. Plasma TAC by FRAP and ORAC assays were negatively correlated with erythrocyte SOD activity. The agreement among the four TAC assay methods and 8-isoprostane was determined using 95% prediction limits of linear regression, expressed as the mean of 8-isoprostane ${\pm}95%$ prediction limits. The ABTS method better agreed with 8-isoprostane than the other methods, demonstrating narrow prediction of limits. Furthermore, only plasma TAC determined by the ABTS assay was inversely correlated with urinary 8-isoprostane (r = -0.35, p < 0.05). In summary, the ABTS assay would be an appropriate method to measure overall plasma antioxidant capacity and predict the body's antioxidant status.

Antioxidant and DNA Damage Protective Activities of Freeze-Dried Blue Mussel (Mytilus edulis) (동결건조 진주담치 추출물의 항산화 및 DNA 손상 보호 활성)

  • Lee, Seon Woo;Choi, Mi-Joo;Kim, Si-Kyung;Lee, Seung-Cheol;Park, Eunju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.12
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    • pp.1801-1807
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    • 2014
  • Blue mussels (Mytilus edulis) are widely distributed among the world's oceans in various habitats. The purpose of this study was to investigate the effects of freeze-drying on the antioxidant and antigenotoxic activities of blue mussels collected in the Gyeongnam coast area of Korea. Raw (RM) and freeze-dried blue mussel flesh (FRM) were extracted with ethanol, methanol, and water. Antioxidant activities were evaluated on the basis of DPPH radical scavenging activity, oxygen radical absorbance capacity (ORAC), cellular antioxidant capacity (CAC), and antigenotoxic activity (comet assay). Except for the water extract, RM and FRM showed DPPH radical scavenging activities, which increased upon freeze-drying in MeOH extract. The highest ORAC value was observed in water extract of RM and MeOH extract of FRM. CAC was protected against AAPH-induced oxidative stress in HepG2 cells by both RM and FRM extracts. Freeze-drying lowered ORAC value of water extract, whereas it increased CAC activity, suggesting that antioxidant activities varied according to the generated radicals. All extracts from RM and FRM showed antigenotoxic activities by reducing $H_2O_2$-induced DNA damage in human leukocytes. Freeze-drying had no effect on antigenotoxicity of blue mussels. Taken together, these results indicate that blue mussels possess antioxidant and antigenotoxic properties, and freeze-drying might be a useful processing method for blue mussels to retain their maximum physiological potential as a functional food.

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 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.

Antioxidant Activity and Safety Evaluation of Juice Containing Protaetia brevitarsis (흰점박이꽃무지(Protaetia brevitarsis) 함유 음료의 in vitro 항산화 관련 생리활성효능 및 안전성 검증)

  • Park, Jae-Hee;Kim, So-Yun;Kang, Min-Gu;Yoon, Min-Soo;Lee, Yang-Il;Park, Eun-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.1
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    • pp.41-48
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    • 2012
  • The purpose of this study was to assess the antioxidant activity of vegetable extracts (pumpkin, aloe, and artichoke) containing Protaetia brevitarsis (PB) and the clinical and pathological changes in ICR mice after a single oral administration. The total polyphenol (TP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging ability, total radical trapping antioxidant potential (TRAP), oxygen radical absorbance activity (ORAC), and single cell gel electrophoresis assay were done to measure their antioxidant activities. The effect of vegetable extracts containing PB in TP and the ORAC value was significantly higher than those without PB. In addition, all extracts had effective $DPPH{\cdot}$ scavenging and $ABTS{\cdot}+$ scavenging activities. The protective effect of vegetable extracts with/without PB on $H_2O_2$-induced DNA damage was found. In a single-dose toxicity study, mortality, body weight, physiological signs, and biochemical analysis were analyzed. Seventy mice were randomly assigned to 7 experimental groups and were administered three vegetable extracts with and without PB (2 g/kg). A full 14 days after administration, no mice mortality was observed in any group. Body weight, physiological signs, and biochemical analysis were never significantly different from those of the control group. Taken together, these findings indicate that vegetable extracts containing PB with antioxidant activities and safety could be applied as medicinal and edible resources in an industrial area.

Enhancement of the Anti-hyperglycemic and Antioxidant Activities of Five Selected Beans by the Germination Process (발아에 따른 콩류의 식후 혈당 상승 억제효능과 항산화 활성)

  • Cho, Cha-Young;Choi, Hwang-Yong;Jo, Sung-Hoon;Ha, Kyoung-Soo;Chung, Ji-Sang;Jang, Hae-Dong;Kwon, Young-In
    • The Korean Journal of Food And Nutrition
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    • v.25 no.2
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    • pp.246-252
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    • 2012
  • After a mixed carbohydrate diet, inhibition of ${\alpha}$-amylase and ${\alpha}$-glucosidase involved in the digestion and absorption of carbohydrates can significantly decrease the postprandial increase of blood glucose level. In the course of screening these useful enzyme inhibitors, we selected five kinds of bean, using an in-vitro enzyme inhibition assay method. To evaluate the effect of germination process on the functionality of the bean, we investigated the inhibitory activities of the water extracts of non-germinated bean and germinated bean against ${\alpha}$-amylase and ${\alpha}$-glucosidase, relevant to postprandial hyperglycemia. We also investigated the oxygen radical absorbance capacity(ORAC), total phenolics content, and postprandial blood glucose lowering effect in rats(Sprague-Dawley rat model). Most germinated beans showed significantly higher ${\alpha}$-glucosidase inhibitory activity, compared with non-germinated beans. Among germinated beans, Glycine max had the highest ${\alpha}$-glucosidase inhibitory activity(53.3%). The water extract of germinated Phaseolus vulgaris L. had the highest ${\alpha}$-amylase inhibitory activity(95.1%), followed by Glycine max(58.7%), and Glycine max L. Merr(54.1%). Furthermore, the five germinated beans also showed high antioxidant activities in ORAC assay. Results suggested that the germination process may improve and enhance the anti-hyperglycemia potential and antioxidant activity of the bean.

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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    • v.18 no.4
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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.

Screening of Traditional Medicines for Antioxidative and Anti-proliferative Effects on Rat Mesangial Cells (한약재 추출물의 항산화 및 사구체혈관간세포 증식 억제활성 탐색)

  • Sohn, Eun-Hwa;Jang, Seon-A;Woo, Han Goo;Koo, Hyun Jung;Han, Hyo-Sang;Kang, Se Chan
    • Korean Journal of Plant Resources
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    • v.26 no.5
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    • pp.652-657
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    • 2013
  • In the present study, anti-oxidative and the RMC proliferation inhibitory propeties of 80% ethanol extracts from 63 kinds of traditional medicines were investigated. Inhibitory effects of RMC proliferation were showed that Dalbergia odorifera T. Chen., Melia azedarach Linn$\acute{e}$ and Hydnocarpus anthelmintica Pierre. Among them Hydnocarpus anthelmintica Pierre had the highest anti-oxidative activity ($ORAC_{PE\;value}=1.6$, DPPH = 81.1), but Dalbergia odorifera T. Chen. and Melia azedarach Linn$\acute{e}$ had no effects. These results suggest that the Hydnocarpus anthelmintica Pierre could prevent or protect from kidney disease as antioxidant and anti-proliferative agent for RMC.

Effect of Red Ginseng Extract on Storage and Antioxidant Activity of Tofu (홍삼 추출물이 두부의 저장성과 항산화 활성에 미치는 영향)

  • Lee, Jung-Suk;Kim, Gyo-Nam;Jang, Hae-Dong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.11
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    • pp.1497-1506
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    • 2008
  • In relation to making tofu with red ginseng extract, the effects of added red ginseng extract on the storage and antioxidant activity of tofu were investigated. When red ginseng extract was added in $0.5%{\sim}2%$ of dried soybean, differences in texture, color and sensory properties were compared. The textural properties of tofu slightly increased depending on the added amount of red ginseng extract in terms of the hardness and springiness: significant difference was demonstrated in the tofu with 2% red ginseng extract, and the color properties also significantly increased in terms of the values of L, a, and b. As a result of sensory evaluation, although the preference to color tended to diminish, as the peculiar taste of red ginseng was significantly perceived, the overall preference did not show any statistical difference. Therefore, the addition of red ginseng extract may be interpreted as not having negative effect on the preference of consumers. There was no change at the early stage of storage in pH, turbidity and total microbial count when the tofu was stored at $15^{\circ}C$. However, the changes in pH and in turbidity, and total microbial counts according to the addition of red ginseng extract took place from the second day and from the sixth day of storage, respectively, suggesting that the shelf-life of tofu may be extended due to its antimicrobial effect. Especially, the highest antimicrobial effect was observed tofu with 2% red ginseng extract. Concerning the antioxidant activity of tofu using oxygen radical absorbance capacity (ORAC) assay, the antioxidant activity significantly increased along with the increase in concentration of red ginseng extract. In ORACOH.assay, the pro-oxidant activity was shown in the tofu added with red ginseng extract.