• Title/Summary/Keyword: DPPH assay

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Antioxidant Activity and DNA Protective Effect against Oxidative Stress of Pinus rigida × taeda Cone (리기테다 소나무 솔방울의 항산화 활성 및 산화적 DNA 손상에 대한 억제 효과)

  • Choi, Jisoo;Jang, Taewon;Min, Youngsil;Lee, Manhyo;Park, Jaeho
    • Journal of Convergence for Information Technology
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    • v.10 no.11
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    • pp.168-176
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    • 2020
  • Reactive oxygen species (ROS) damage DNA and cause cancer. Therefore, the research is being conducted on the development of antioxidants for the removal of ROS. This study was performed to investigate antioxidant activity and protective effect against oxidative DNA damage using ethyl acetate fractions from the cone of Pinus rigida × taeda (ERT). The antioxidant activity was evaluated using the DPPH, ABTS radical scavenging assay, reducing power assay, and Fe2+ chelating assay. Also, the contents of phenolic compounds and vitamin C related to antioxidant activity were analyzed to confirm phytochemicals. The DNA protective effect against oxidative stress was confirmed by the φX-174 RF I plasmid DNA cleavage assay. As a result, ERT showed DPPH and ABTS radical scavenging activities in a concentration-dependent manner. The results of reducing power and Fe2+ chelating activities were 77.32 ± 2.28% and 64.09 ± 1.01% at 200 ㎍/㎖. Also, ERT showed a DNA protective effect against oxidative stress.

Analysis of Antioxidative Activity and Antimutagenic Effect of Ethanol Extract from Schizandra chinensis Baillon (오미자(Schizandra chinensis Baillon) 에탄올 추출물의 항산화 활성 및 항돌연변이 활성 분석)

  • Jeon, Yeon-Bee;Kil, Jin-Bee;Lim, So-Min;Kim, Mee-Hyun;Kim, Mee-Ra
    • Journal of the East Asian Society of Dietary Life
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    • v.18 no.5
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    • pp.746-752
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    • 2008
  • In this study, the antioxidative activity and antimutagenic effect of ethanol extracts from Schizandra Chinensis Baillon were assessed with regard to natural antioxidants. The antioxidative activity and antimutagenicity of ethanol extracts from Schizandra chinensis Baillon were evaluated by measuring the radical-scavenging effect on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, and by performing the Ames test using 4-nitroquinoline-1-oxide (4-NQO) and sodium azaid as a mutagen with Salmonella typhimurium TA100, respectively. The total polyphenolic compound and flavonoid compound contents of Schisandra chinensis Baillon were also assessed. The DPPH radical-scavenging activity of ethanol extracts from Schisandra chinensis Baillon was 57.2% on the $500\;{\mu}g$/assay. The $IC_{50}$ on DPPH radical-scavenging effect of ethanol extract was $435\;{\mu}g$/assay. In addition, the inhibition rates of ethanol extract from Schisandra chinensis Baillon toward mutagenicity induced by 4-NQO and sodium azide were 82.45% and 45.3%, respectively. Furthermore, the total polyphenol and total flavonoid contents of the extract were 9.53 mg/g and 3.97 mg/g, respectively. These results indicate that the ethanol extract from Schisandra chinensis Baillon evidences a fairly good antioxidative and antimutagenic effect.

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Antioxidant Activity and Inhibitory Effect on Oxidative DNA Damage of Ethyl Acetate Fractions Extracted from Cone of Red Pine (Pinus densiflora) (솔방울 에틸아세테이트 분획물의 항산화 및 산화적 DNA 손상 억제 활성)

  • Jang, Tae Won;Nam, Su Hwan;Park, Jae Ho
    • Korean Journal of Plant Resources
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    • v.29 no.2
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    • pp.163-170
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    • 2016
  • Antioxidant activity and inhibitory effect on oxidative DNA damage of ethyl acetate fractions extracted from Cone of Red Pine (Pinus densiflora) were investigated to find utilization of Cone, by-product of Red Pine, thrown out after berry shatter, as a new natural plant resource. Cone from P. densiflora was extracted with methanol (MeOH) and separated to petroleum ether, ethyl acetate and water fraction. Among them, ethyl acetate fraction was used. The antioxidant activity was conducted by the 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical, 2, 2'-Azino-bis (3-ethylbenzothiazoline-6 sulfonic acid) diammonium salt (ABTS) radical scavenging assay, Fe2+ chelating assay and reducing power assay. The inhibitory effect on oxidative DNA damage was determined by DNA cleavage assay using φX-174 RF I plasmid. The results of DPPH and ABTS radical scavenging activity at 200 ㎍/㎖ of extracts were 86.50% and 95.80% respectively, which were similar figures compared with L-ascorbic acid as control. Fe2+ chelating activity was 77.96% and reducing power was 0.77 at 200 ㎍/㎖. Total phenolic component was 27.29±0.3 ㎎/g and Vitamin C content was 1.84±0.1 ㎎/g. Also ethyl acetate fraction from Cone has inhibitory effect, using φX-174 RF I plasmid on DNA cleavage assay. In conclusion, Cone, by-product of P. densiflora, showed high antioxidant activity and inhibitory effect on oxidative DNA damage. Therefore this study suggests Cone, useless by-product, can be developed as a new natural plant resource with lots of utilization such as an effective antioxidant, natural medicine, food, cosmetics and so on.

Antioxidant Activities of Rubus coreanus Miquel and Morus alba L. Fruits (복분자와 오디의 항산화 특성)

  • Jun, Hyun-Il;Kim, Young-Ae;Kim, Young-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.3
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    • pp.381-388
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    • 2014
  • The antioxidant activities of extracts from Rubus coreanus Miquel (black raspberry) and Morus alba L. (mulberry) fruits were investigated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, 2,2-azino-bis-(3-ethylbenzo-thiazoline-6-sulfonic acid) (ABTS) assay, and reducing power assay. Aqueous mixtures of ethanol, methanol, and acetone were analyzed in order to determine the most effective extraction solvent for the two fruits. Black raspberry and mulberry extracts with the 60:40 acetone-water mixtures (v/v) showed the highest DPPH radical scavenging activities (56.2 and 85.2%, respectively) compared to the other extraction solvents. The 60% acetone extract was finally selected as an extraction solvent and then sequentially fractionated according to solvent polarity. Among the fractions of the two fruits, the ethyl acetate fraction showed the highest antioxidant activity as well as total phenolic and total flavonoid contents. In addition, there were high correlation coefficients between antioxidant activities and their contents. The $EC_{50}$ value of the ethyl acetate fraction from mulberry fruit was 2.2 times lower than that of butylated hydroxytoluene (BHT) in DPPH assay. The major phenolic acid and anthocyanin of the two fruits were protocatechuic acid and cyanidin-3-glucoside, respectively.

Comparison of the Total Phenolic and Flavonoid Contents and Antioxidant Activities of Four Kinds of Sand Dune Plants Living in Taean, Korea (태안에서 서식하는 사구식물 4종의 폴리페놀, 플라보노이드 함량과 항산화 활성 비교)

  • Kim, Soon-seok;Cha, Hyeon-cheol
    • Korean Journal of Plant Resources
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    • v.30 no.1
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    • pp.8-16
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    • 2017
  • This study attempted to investigate the antioxidant properties of four different species of sand dune plants (Calystegia soldanella, Messerschmidia sibirica, Vitex rotundifolia and Rosa rugosa). In order to validate the antioxidant activity of these plants, we first determined the total amount of flavonoid versus phenolic contents (TFC/TPC) and extracted crude flavonoids for measuring antioxidant activities that were determined by DPPH, ABTS and FRAP assays with radical scavenging effects. We found that highest amounts of TPC were detected in R. rugose with values of 110.20 mg/g (leaves) and 65.71 mg/g (stems), while the highest amounts of TFC in V. rotundifolia with values of 38.07 mg/g (leaves) and 6.55 mg/g (stems). We further examined how closely related the amounts of TFC/TPC in antioxidant activities and found that R. rugose has the highest activities of radical scavenging with values of $63.4{\mu}g/ml$ and $51.2{\mu}g/ml$ determined by DPPH and ABTS assays compared with the value of $21.2{\mu}g/ml$ by FRAP assay. It is of note that there is a statistically significant correlation between the resulting antioxidant activities and the total ratio of TFC and TPC, suggesting that the different amounts of TFC/TPC may directly contribute to the various antioxidant activities.

Effect of Antioxidant Activity and Induction of DNA Damage on Human Gastric Cancer Cell by Rubus coreanus Miquel (복분자 추출물의 항산화활성 및 인간 위암 세포주에 대한 유전적 손상 유도)

  • Jeon, Sang-Kyung;Lee, Ji-Won;Lee, In-Seon
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1723-1728
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    • 2007
  • Rubus coreanus Miquel (RCM), a type of red raspberry, grows wild in Korea and China and its unripe fruit is used as a folk medicine for the treatment of impotence and as a diuretic. RCM was extracted with methanol and then further fractionated it into for different types. In this study, we investigated the antioxidant activity of a RCM extract (ext.) and its fraction (fr.). DPPH free radical scavenging activity assay, total polyphenols contents, total flavonoids contents assay were used to analyze antioxidant activity. The DPPH free radical scavenging activity $(RC_{50}:1.67{\mu}g/ml)$ and total polyphenols contents $(546.25{\mu}g/mg)$ were higher in butanol fraction than in other fr. And total flavonoids contents was higher in ethylacetate fr. $(141.78{\mu}g/mg)$. We applied comet assay to measure the DNA damage in the individual cells and exposed time course at $IC_{50}$. Comet assay is a rapid and sensitive fluorescent microscopic method to examine DNA damage and repair at individual cell level. The butanol fro from RCM significantly induced 54.12%, 57.95% of DNA damage after treated RCM for 8 hr. In conclus

Purification of Antioxidant substance from the stem bark of Rhus verniciflua

  • Kim, Jung-Bae
    • Proceedings of the Korean Journal of Food and Nutrition Conference
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    • 2001.12a
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    • pp.126-126
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    • 2001
  • The Rhus verniciflua contains alkly(en)-catechol type allergens with a saiurated or unsaturated alkly chain of 15 or 17 carbon atoms. It has been recognized as an extremely active allergen causing skin reactions similar In poison ivy. The allergic contact dermatitis induced by the urushiol is known to be mediated be T lymphocytes whicht specifically recognize the hepten urushiol. Therefore. direct use of this plant as a medicinal purpose might imply a considerable hazard in Korea. In this study, using the established method for the detoxification from the stem bark of Rhus verniciflua, an strong antioxidant substance was isolated and characterized DPPH (diphenypricryl hydrazyl) assay measures hydrogen atom-donating activity and hence provides a measure of free radical scavenging antioxidant activity. DPPH, a purple-colored stable free radical, is reduced to yellow-colored diphenylpicryl hydrazine by antioxidants to deducing agents. Antioxidative effects of the water extract from RV were measured by DPPH assay. Twenty microliters of the extract was added to 1ml of 100mM DPPH solution in ethanol The mixture was shaken and left to stand for 10min at room temperature. The crude water extracts was purified by using HPLC method with a DEAE (anionic type), CN, ODS column. The purified compound remained stable at pH 3.0-6,0, but unstable above pH 6.5. It was stable heat at 10$0^{\circ}C$ for 4 hours, but still had about 80% of residual activity after treatment at 10$0^{\circ}C$ for 5 hours. The elemental composition of the HR-EI mass spectrum at m/z 170.02 was estimated the empirical formula as $C_{7}$ $H_{6}$ $O_{5}$. $C_{10}$ $H_4$ $O_2$N$_1$, $C_{5}$ $H_4$ $O_4$N$_3$, $C_{8}$$H_2O$$_1$N$_4$. In antimicrobial test, no inhibition was observed against Gram-positive and negative bacteria. This compound was stronger than that of commercial antioxidant by DPPH test, such as BHT, BHC at the same concentration (20$\mu$g/ml).ml).

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Antioxidant Activities of Bacterial Culture Extracts Isolated from Arctic Lichens (북극 지의류로부터 분리한 미생물 배양 추출액의 항산화 활성)

  • Kim, Mi-Kyeong;Park, Hyun;Oh, Tae-Jin
    • Microbiology and Biotechnology Letters
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    • v.40 no.4
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    • pp.333-338
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    • 2012
  • Lichens are a symbiosis between fungi, algae and cyanobacteria. Our group recently studied the antioxidant properties of some bacterial species isolated from Arctic lichens and we confirmed that they possess high antioxidant activities. In this paper, we investigated the antioxidant capacity of 5 microorganisms newly isolated from 4 Arctic lichen species, Cladonia sp., Sterocaulon sp., Umbilicaria sp. and Cetraria sp., using various solvent extractions. We carried out 1,1-diphenyl-2-picryl-hydrazyl (DPPH) and 2,2'-azino-bis(3-ethyl benzothiazoline-6-sulphonic acid) (ABTS) free radical scavenging activity test and ferric reducing antioxidant power (FRAP) assay. Also total phenolic and flavonoid content assays were performed. Among the bacterial culture extracts of the tested lichen-microorganisms, ethyl acetate extracts of Burkholderia sordidicola S5-B(T) had not only a high antioxidant activity (72.9%) when compared with the ascorbic acid used as the control (51.3%) in the DPPH assay, but also a high amount of phenolic content as well as flavonoid content. As a result, these lichen-microorganisms may be potentially useful sources of natural antioxidants.

Potential of Wood Vinegar in Enhancing Fruit Yield and Antioxidant Capacity in Tomato

  • Benzon, Hiyasmin Rose L.;Lee, Sang Chul
    • Korean Journal of Plant Resources
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    • v.29 no.6
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    • pp.704-711
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    • 2016
  • Tomatoes are considered as one of the main components of daily meals in most households. Thus, it is important to invest in studies enhancing their yield and nutritional value. The study evaluated the effect of wood vinegar (WV) on tomato under greenhouse conditions. Data on fruit number, fruit weight, and plant height were recorded. Total phenolic content (TPC) and antioxidant activity of tomato were determined using the Folin-Ciocalteu reagent and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, respectively. FRR-CF+500x-WV significantly increased the fruit number (86.11%) and fruit weight (81.78%) over the control. Results were comparable to HRR-CF+250x-WV, indicating that conventional fertilizer and WV may have synergistic effect on each other. TPC and scavenging effect on DPPH radical was improved by 250x-WV application. The increase in the phenolic compounds can be attributed to WV as a direct source or as a factor triggering the plants to produce more secondary metabolites with the purpose of increasing natural defenses. The significant effect obtained by applying the full recommended rate of conventional fertilizer on the carotenoid content was due to the availability of the major nutrients needed by the plant. Correlation analysis showed that carotenoids have negative correlation with TPC and DPPH. However, TPC and DPPH showed that these are positively correlated with each other. Tomatoes are excellent source of antioxidants associated with the reduction of some human diseases and improved health. The results provided evidence that WV alone and/or its combination with conventional fertilizers has favorable effects on the quality of tomato.

In Vitro Antioxidant Activity Profiles of ${\beta}$-Glucans Isolated from Yeast Saccharomyces cerevisiae and Mutant Saccharomyces cerevisiae IS2

  • Song, Hee-Sun;Moon, Ki-Young
    • Food Science and Biotechnology
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    • v.15 no.3
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    • pp.437-440
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    • 2006
  • To explore the possible usefulness of ${\beta}$-glucans as natural antioxidants, the antioxidant profiles of ${\beta}$-glucan, extracted from Saccharomyces cerevisiae KCTC 7911, and water soluble and insoluble mutant ${\beta}$-glucan, isolated from yeast mutant S. cerevisiae IS2, were examined by five different in vitro evaluation methods: lipid peroxidation value (POV), nitric oxide (NO), 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, reducing power, and ${\beta}$-carotene diffusion assay. The antioxidant activities of all ${\beta}$-glucans evaluated in POV test were comparable to or better than that of the known antioxidant, vitamin C. Remarkably, the ${\beta}$-glucan and water insoluble mutant ${\beta}$-glucan possessed 2.5-fold more potent activity than vitamin C at a dosage of 2 mg. Although vitamin C showed 100-fold greater activity than all ${\beta}$-glucans in NO and DPPH tests for measuring the radical scavenging capacity, all ${\beta}$-glucans revealed higher radical scavenging activity than the known radical scavenger, N-acetyl-L-cysteine (NAC), in DPPH test. The water insoluble mutant ${\beta}$-glucan had 2.6- and 5-fold greater antioxidative activity than water soluble ${\beta}$-glucan in NO and DPPH tests, respectively, showing that all ${\beta}$-glucans were able to scavenge radicals such as NO or DPPH. While all ${\beta}$-glucans revealed lower antioxidant profiles than vitamin C in both reducing power activity and ${\beta}$-carotene agar diffusion assay, the ${\beta}$-glucan and water insoluble mutant ${\beta}$-glucan did show a marginal reducing power activity as well as a considerable ${\beta}$-carotene agar diffusion activity. These results confirmed the potential usefulness of these ${\beta}$-glucans as natural antioxidants.