• Title/Summary/Keyword: Antioxidant assay

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Protective Effect of Aqueous Extract from Erigeron annuus Against Cell Death Induced by Free Radicals

  • Myeongguk Jeong;Hyeokjin Kwon;Youngdon Ju;Go-Eun Choi;Kyung-Yae Hyun
    • Biomedical Science Letters
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    • v.28 no.4
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    • pp.307-311
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    • 2022
  • The extract of EA lacks studies showing its efficacy other than that it contains caffeic acid, an active compound that has antioxidant and neuroprotective effects on nerve cells. Therefore, in this study, we attempted to determine the effectiveness of EA extraction. In this study, we performed a DPPH assay to determine the antioxidant potential of EA. And then, the cytotoxic concentration of EA in HaCaT keratinocytes was determined, and the antioxidant effect was determined by measuring the malondialdehyde (MDA). The results of DPPH, a chemical antioxidant assay, clearly demonstrated the antioxidant capacity of EA extracted with distilled water. In addition, cell-based assays provide useful information on the protective effect of EA on oxidative stress-induced apoptosis.

In Vitro Antioxidant Activity of Alnus firma Extracts (사방오리(Alnus firma) 추출물의 in vitro 항산화 활성)

  • Choi, Hye Jung;Joo, Woo Hong
    • Journal of Life Science
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    • v.29 no.2
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    • pp.231-238
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    • 2019
  • This study evaluated the antioxidant activity of the extract and fractions of Alnus firma. Alnus firma had the highest total phenolic content ($452.80{\pm}7.01{\mu}g$ gallic acid equivalents/mg) in a methanol (MeOH) fraction and the highest total flavonoid content ($112.29{\pm}11.14{\mu}g$ rutin equivalents/mg) and antioxidant capacity ($936.23{\pm}0.07{\mu}g$ ${\alpha}$-tocopherol equivalents/mg) in an ethylacetate (EA) fraction. The antioxidant activities of various solvent extract fractions of Alnus firma were evaluated using various antioxidant assays, including ${\beta}$-carotene-linoleate assay, reducing power assay, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, metal chelating activity assay, superoxide anion radical scavenging assay, and lipid peroxidation inhibition assay using the ferric thiocyanate method. These activities were compared with those of ascorbic acid, butylated hydroxyanisole (BHA), gallic acid (GA), butylated hydroxytoluene (BHT), and ${\alpha}$-tocopherol. First, at a $250{\mu}g/ml$ concentration, the EA and MeOH fractions of A. firma showed 92.43% and 89.20% DPPH radical scavenging activity, respectively. Second, $50{\mu}g/ml$ of the EA fraction exhibited 72.49% superoxide anion radical scavenging activity, a little greater than the same dose of GA (60.88%). Finally, 0.5 and 1 mg/ml of the EA fraction showed 73.45% and 73.29% inhibition of peroxidation in the ${\beta}$-carotene-linoleic acid system, respectively. The decreasing order of reducing power was EA fraction > n-butanol (BuOH) fraction > dichloromethane (DCM) fraction > n-hexane (HX) fraction. The results obtained in the present study indicated that Alnus firma can be used as an easily accessible potential source of natural antioxidants.

Protective Effect of Co-treatment of Lutein and Fucoidan Against AAPH-Induced Damage in THP-1 Cells (루테인과 후코이단 병용 처리에 의한 AAPH 유도 세포 손상 억제)

  • Lee, Keyong-Ho;Yoon, Won-Ho
    • The Korean Journal of Food And Nutrition
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    • v.23 no.3
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    • pp.306-310
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    • 2010
  • This study was designed to investigate the protective effect of the combination of fucoidan and lutein against AAPH-induced oxidative stress in THP-1 cells. The combination of fucoidan and lutein existed significant antioxidant effect on AAPH-damaged THP-1 cells by using lipid peroxidation and cellular antioxidant capacity assay. Fucoidan($1\;{\mu}g/m{\ell}$) and lutein($10\;{\mu}g/m{\ell}$) did not affect at all the viability of THP-1 cells, but protected the AAPH-damage of THP-1 cells at the same concentration. The viability of THP-1 cells was 0% with 1 mM AAPH alone, the protective effect of fucoidan($1\;{\mu}g/m{\ell}$) and lutein($10\;{\mu}g/m{\ell}$) was 37% and 36%, respectively. The combination of fucoidan($1\;{\mu}g/m{\ell}$) and lutein($10\;{\mu}g/m{\ell}$) exhibited significant inhibitory effect of lipid peroxidation using TBARS assay and cellular antioxidant capacity using DCFH-DA assay. In lipid peroxidation, the TBARS value of 1 mM AAPH alone was $0.8{\pm}0.03\;nM$ MDA, its of the combination of fucoidan($1\;{\mu}g/m{\ell}$) and lutein($10\;{\mu}g/m{\ell}$) was $0.2{\pm}0.05\;nM$ MDA. In cellular antioxidant capacity, the combination of fucoidan($1\;{\mu}g/m{\ell}$) and lutein($10\;{\mu}g/m{\ell}$) exhibited significant cellular antioxidant capacity of 76%, whereas quercetin($10\;{\mu}M$) as positive control exhibited the cellular antioxidant capacity of 32%. These results indicate that the cotreatment of fucoidan and lutein protects against AAPH-induced THP-1 cell damage by inhibiting lipid peroxidation, increasing cellular antioxidant capacity.

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.

Antioxidant and Neuronal Cell Protective Effect of Purple Sweet Potato Extract (자색고구마 추출물의 항산화 효과 및 신경세포 보호효과)

  • Kwak, Ji-Hyun;Choi, Gwi-Nam;Park, Ju-Hee;Kim, Ji-Hye;Jeong, Hee-Rok;Jeong, Chang-Ho;Heo, Ho-Jin
    • Journal of agriculture & life science
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    • v.44 no.2
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    • pp.57-66
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    • 2010
  • The antioxidant and neuronal cell protective effects of water extract from purple sweet potato were investigated. The total phenolics and monomeric anthocyanin contents of purple sweet potato extract were 44.25 mg/g and 2,394 mg/L, respectively. The antioxidant activities of purple sweet potato extract were evaluated using various antioxidant tests, including 1,1-diphenyl- 2-picrylhydrazyl (DPPH), 2,2'-azino- bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging activities, ferric reducing/antioxidant power (FRAP) and reducing power. In these assays, the extract of purple sweet potato presented significant radical scavenging activities, FRAP, and reducing power in a dose-dependent manner. MTT {3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl- tetrazoliumbromide} reduction assay showed significantly increase in cell viability when PC12 cells were pretreated with purple sweet potato extract. Because oxidative stress is also known to increase neuronal cell membrane breakdown, we further investigated by lactate dehydrogenase (LDH) and neutral red uptake assay. Purple sweet potato extract inhibited oxidative stress-induced membrane damage in neuronal cells. Therefore, these data results demonstrated that water extract of purple sweet potato have antioxidant activity and neuronal cell protective effect thus it has great potential as a natural source for human health.

Analysis of Antioxidant Activity and Cytotoxicity against Human Cancer Cell Lines of Extract from Atractylodes rhizoma fermented with Ganoderma lucidum Mycelium (창출과 영지버섯 균사체 발효 추출물의 항산화 활성 및 인체 암세포주에 대한 세포독성 분석)

  • Park, Mi Hye;Kim, Mee Ra
    • The Korean Journal of Food And Nutrition
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    • v.30 no.3
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    • pp.454-463
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    • 2017
  • Ganoderma lucidum has been traditionally used as a medicine for treatment of bronchitis, arthritis, and high blood pressure, and it has been reported to display many biological activities including anticancer and immune activities. Since mushroom mycelium is known to have excellent biological activities together with mushroom fruiting body, studies on biological activities of mushroom mycelium have been actively conducted. Thus, the present study compared the biological activities before and after the cultivation of Ganoderma lucidum mycelium on Atractylodes rhizoma. When the radical scavenging activity was assessed by the DPPH assay, ARGL (ethanol extract of Atractylodes rhizoma mycelium fermented with Ganoderma lucidum) showed radical scavenging activity of 5.58~82.56% at concentrations of $10{\sim}500{\mu}g/assay$, while AR (ethanol extract of Atractylodes rhizoma) showed radical scavenging activity of 5.27~72.08% at the same concentrations. When measured by using the ABTS assay, ARGL showed higher radical scavenging activity than AR, which was consistent with the result obtained by the DPPH assay. In the MTT assay, the cytotoxicity of ARGL against all cell lines was higher than that of AR. In particular, the cytotoxicities of AR and ARGL against Hep3B at a concentration of $400{\mu}g/assay$ were 71.81% and 86.40%, respectively. In addition, the result obtained by the SRB assay was consistent with the result obtained by the MTT assay. According to the results mentioned above, there is a high probability that medicinal herb cultures using mycelium can be used as sources of functional foods since the cytotoxicities against cancer cells and antioxidant activities increased when the mycelium was fermented with Atractylodes rhizoma.

Evaluation of Antioxidant Activity of Sugar Alcohols Using TOSC (Total Oxy-radical Scavenging Capacity) Assay (TOSC 법을 이용한 당알코올의 항산화 활성 평가)

  • Kang, Keon-Wook;Kwak, Sang-Hoon;Yun, Sei-Young;Kim, Sang-Kyum
    • Toxicological Research
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    • v.23 no.2
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    • pp.143-150
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    • 2007
  • Although animal and epidemiological studies have suggested oxidative stress as an etiological factor in pathogenesis including cancer, inflammation, sepsis, fibrosis, cardiovascularlneurodegenerative diseases and aging-related disorders, conflicting results have been obtained in clinical trial with antioxidants. The reason for this discrepancy remains unknown but may be due, in part, to the lack of a validated assay system for evaluating antioxidant capacity. The antioxidant activity of a series of sugar alcohols against peroxyl radicals, hydroxyl radicals and peroxynitrites was determined by the total oxy-radical scavenging capacity (TOSC) assay and cell-based assay using H4IIE cells. Specific TOSC values calculated from the slope of the linear regression for erythritol, xylitol, sorbitol or mannitol against peroxyl radicals was $2.1{\pm}0.2,\;3.7{\pm}0.3,\;9.1{\pm}0.3$ or $8.7{\pm}1.1$ TOSC/mM, respectively. Specific TOSC values for erythritol, xylitol, sorbitol or mannitol against peroxynitrite was $1.9{\pm}0.3,\;3.9{\pm}0.4,\;7.8{\pm}0.7$ or $7.7{\pm}0.5$ TOSC/mM, respectively. These results suggest that oxy-radical scavenging capacity is dependent on the number of aliphatic hydroxyl group in sugar alcohols of monosaccharide. Tert-butylhydroperoxide (t-BHP)-induced cell toxicity determined by MTT assay was marginally attenuated by 10 mM erythritol, but completely inhibited by 10 mM xylitol, 2 mM sorbitol or 0.75 mM maltitol, a disaccharide alcohol. Oxidative stress markers, such as glutathione (GSH) and malondial-dehyde (MDA) levels, were measured in t-BHP-treated cells using HPLC equipped with a fluorescence detector and a reverse phase column. Erythritol did not change the levels of GSH and MDA in H411E cells treated with t-BHP. The t-BHP-induced changes in cellular GSH and MDA levels were ameliorated by 10 mM xylitol and completely blocked by 10 mM sorbitol and maltitol. These results indicate that sugar alcohols protect cells against oxidative stress via scavenging oxy-radical and suggest that TOSC assay in conjunction with cell-based assay is a valid method for evaluating antioxidant capacity of natural and synthetic chemicals.

Antioxidant Activity and Neuroprotective Effect of Psoralea corylifolia Linne Extracts (파고지 (Psoralea corylifolia Linne) 추출물의 항산화 작용에 의한 신경세포 보호효과)

  • Yoon, Mi-Young;Lee, Bo-Bae;Kim, Ju-Young;Kim, Yong-Seong;Park, Eun-Ju;Lee, Seung-Cheol;Park, Hae-Ryong
    • Korean Journal of Pharmacognosy
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    • v.38 no.1
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    • pp.84-89
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    • 2007
  • The present study describes the preliminary evaluation of the antioxidant activities and the neuroprotective effect of methanolic extracts from Psoralea corylifolia Linne (PCE). The antioxidant activities and neuroprotective effect of the PCE were evaluated by total phenolic contents (TPC), DPPH radical scavenging activity (RSA), reducing power (RP), MTT reduction assay, and LDH release assay. TPC, DPPH RSA, and RP of the extract at concentration of 100 ${\mu}g$ was 125.93 ${\mu}g$, 63.81%, 0.138, respectively, and those were concentration dependent. The treatment of PC12 and N18-RE-105 cells with various PCE concentrations under $H_2O_2$ resulted in the induction of protective effect in a dose-dependent manner, as determined by the results of an MTT reduction assay and LDH release assay. Therefore, these results suggest that PCE could be a new potential candidate as an antioxidant against neuronal diseases.

Fluctuations in Phenolic Content and Antioxidant Capacity of Green Vegetable Juices during Refrigerated Storage

  • Kim, Seong Yeong
    • Preventive Nutrition and Food Science
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    • v.20 no.3
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    • pp.169-175
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    • 2015
  • Shinseoncho and kale were made into green vegetable juices by building block [shinsenocho branch (SB), shinsenocho leaf (SL), kale branch (KB), and kale leaf (KL)]. Fluctuations in their phenolic contents and antioxidant capacities were analyzed during refrigerated storage at $4^{\circ}C$ for 28 days. Total polyphenolic contents of leaf parts showed a decreasing tendency after 4 days (SL) or 7 days (KL), whereas branch parts showed fluctuating values during the entire storage period. The 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging capacity was rapidly decreased in SB and in SL at 28 days (P<0.001), whereas KL showed a slightly increasing tendency after 14 days. For the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity, SL showed a sharp fall at 28 days (P<0.001), and KL showed a decreasing tendency after 14 days (P<0.001). SB showed a steady decrease during the entire storage period and KB indicated a nearly zero (0.97%) at 28 days. Pearson's coefficients for the correlation between antioxidant capacities measured by the ABTS and DPPH assays, and the total polyphenolic contents were determined. The results showed that the ABTS assay (r=0.934, P<0.001) was more strongly positively correlated with the total phenolic contents than the DPPH assay (r=0.630, P<0.001). In conclusion, when considering all building blocks, green vegetable juices, including kale and shinseoncho may have kept antioxidant capacities for up to 14 days under refrigeration, and the ABTS assay better reflects a positive correlation with the total phenolic contents when compared to the DPPH assay.

Evaluation of Antioxidant and Anticancer Activity of Guarana and Graviola in Human Colon Cancer (과라나와 그라비올라가 인체 대장암에 미치는 항산화 및 항암효과에 대한 연구)

  • Lee, Myeong-Seon
    • Journal of Korea Entertainment Industry Association
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    • v.13 no.1
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    • pp.217-223
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    • 2019
  • Colon cancer is the most common form of cancer diagnosis in worldwide. There are growing interests in the health benefits associated with consumption of fruits and vegetables, especially for the prevention of cancer, cardiovascular or other chronic diseases. The objective of the present study was to investigate the antioxidant and anticancer activities of natural product, guarana(GR) and graviola(GV) in human colon carcinoma HCT-116 cells. MTT assay, flow cytometry analysis were employed to investigate the anticancer mechanism and DPPH assay was determined to the antioxidant activity to scavenge free radicals in extract of these. All two extracts showed significantly antioxidant activity at 50mg/ml of concentration. GR and GV reduced HCT-116 cell proliferation in a dose dependent manner. Specially GR treatment(96.65±3.71) also significantly increased the sub-G1 population more than GV(79.58±2.87) treatment in HCT-116 at the concentration of 10mg/ml, as shown by flow cytometry assay. Statistical analyses revealed GR and GV exhibited significantly high (P < 0.05) cytotoxicity in HCT-116. These findings indicate that GN and GV may serve as novel therapeutic agents for colon cancer treatment and future leads for drug development.