• Title/Summary/Keyword: DPPH Assay

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Chemical changes in resazurin by probiotics and its application for evaluating living bacterial cell counts and their reduction potentials (프로바이오틱스에 의한 레자주린의 화학적 변화와 생균수 및 환원활성 측정에의 적용)

  • Lee, Hyowon;Oh, Yeong Ji;Hong, Jungil
    • Korean Journal of Food Science and Technology
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    • v.53 no.2
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    • pp.204-212
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    • 2021
  • Resazurin, an oxidized blue dye, is reduced to resorufin, showing a peak absorbance change and emitting fluorescence due to the metabolic activity of living cells. In this study, the growth of Lactobacillus rhamnosus GG (LGG) and the redox potential of living probiotic bacteria were evaluated based on changes in the absorbance or fluorescence of resazurin. Fluorescence analysis is a more accurate and sensitive method for quantifying viable LGG than the colorimetric absorbance measurement of resazurin change. Fluorescence measurement could detect LGG of 6.5-9.5 log CFU/mL within 30 min with R2=0.99. No significant effect of further reduction of resorufin to dihydroresorufin by LGG was observed. Various probiotics showed different resazurin-reducing activities, and L. kimchicus had the highest reducing activity among the six probiotics tested. These findings suggests that fluorescence measurement in a resazurin-based assay is useful for analyzing bacterial growth and the redox potential of living probiotics.

Physiological activities of leaf extract of Lonicera morrowii A.Gray, a plant native to Ulleungdo (울릉도 자생식물인 섬괴불나무(Lonicera morrowii A.Gray) 잎 추출물의 생리활성)

  • Lee, Eun-Ho;Park, Hye-Jin;Hong, Eun-Jin;Akhmadjon, Sultanov;Kim, Byung-Oh;Jung, Hee-Young;Kang, In-Kyu;Cho, Young-Je
    • Journal of Applied Biological Chemistry
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    • v.63 no.4
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    • pp.443-449
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    • 2020
  • This study was performed to investigate total phenolic contents, anti-oxidant, biological disease-related enzyme inhibitory, and anti-microbial effects of extracts of Lonicera morrowii leaves prepared with water and 40% ethanol. Anti-oxidative activities of the extracts increased in a dose-dependent manner and were very high even at low phenolic concentration. At phenolic concentrations ranging between 50 and 200 ㎍/mL, the water and ethanol extracts inhibited 39.34-76.35 and 47.53-99.83% of xanthine oxidase activity, 30.21-79.06, and 59.40-87.14% of angiotensin converting enzyme activity, and 59.81-80.35 and 36.06-63.58% of α-glucosidase activity, respectively. All inhibitory effect were increased in a dose-dependent manner. In the paper disc agar diffusion assay against seven microbes, L. morrowii extracts had anti-bacteria activities against Helicobacter pylori, Propionibacterium acne but no effect against other bacteria. These results indicate that the extracts from L. morrowii leaves have the possibility to be developed as a physiologically functional source for prevention of adult diseases.

Antioxidant, Anti-inflammatory and Anti-photoaging Activities of Hydrolyzed Peony (Paeonia lactiflora Pall.) Flower (작약꽃 가수분해물의 항산화, 항염 및 광노화 억제 효능)

  • Kyung Ju Lee;You Ah Kim;Byoung Jun Park
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.49 no.3
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    • pp.259-267
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    • 2023
  • This study was conducted to evaluate physiological activity of flower extract of peony (Paeonia lactiflora Pall.) by hydrolysis and to use it as a valuable cosmetic ingredients. Four cultivar of peony flowers were extracted, and the highest active ingredient contents was selected, and that cultivar was used for hydrolyzing. The results showed that high concentration of hydrochloric acid (HCl) hydrolyzed, and biological hydrolysis using enzymes had no activity. The deglycosylation of peonidin 3,5-diglucoside occurred by hydrolysis. The hydrolysate contains 63.3 ppm of peonidin, a red-colored anthocyanin compound. The antioxidant activity of hydrolysate was compared with extract. The results showed the strong antioxidant activity in hydrolysate (96%) than extract (82%). In addition, hydrolysate of peony flower showed higher inhibitory activity of NO release than extract. UVA assay using fibroblast cell (CCD-986Sk) showed that hydrolysate increased cell viability than extract under UVA exposure. Based on these results, we anticipate that hydrolysate of peony flower can be used a valuable cosmetic ingredient.

Comparison of Antioxidant, Antidiabetic, and Antithrombotic Activities of Native Korean and Improved Pepper Varieties (국내 재래종 고추 및 개량종 고추의 항산화, 항당뇨 및 항혈전 활성의 비교)

  • Eun-Seo Lim;Seong-Im Park;Jong-Sik Kim;Ho-Yong Sohn
    • Journal of Life Science
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    • v.34 no.6
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    • pp.385-392
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    • 2024
  • Hot pepper (Capsicum annuum L.) is a valuable plant that is widely used worldwide for food and medicinal purposes. This study compared ethanol extracts of five native Korean varieties (Yuwol-cho, Subi-cho, Sumihyang, Gounbit, and Chilseong-cho) and five improved varieties (Dabok, Cheongyang, Chungseong, Olbokhap, and Shin-honggildong) of peppers cultured in Korea. The extracts were analyzed for color difference, polyphenol content, and their antioxidant, antidiabetic, and antithrombotic activities. The extracts of the improved varieties exhibited higher levels of redness and lower levels of yellowness compared to the native varieties. Polyphenol and flavonoid content analysis revealed significantly higher levels in the Yuwol-cho and Sumihyang varieties, which also exhibited scavenging activities in 1,1-diphenyl-2-picryl hydrazyl anion-, 2,2-azobis(3-ethylbenzothiazoline-6-sulfonate) cation-radical scavenging, and reducing power assay. The Chilseong-cho, Yuwol-cho, and Dabok varieties showed significantly higher nitrite scavenging activity. Antidiabetic activity based on α-glucosidase inhibition was observed in the Subi-cho, Sumihyang, and Gounbit extracts. Evaluation of antithrombotic activity showed that the Yuwol-cho extract prolonged thrombin time by 1.61 times compared to the solvent control at a concentration of 5 mg/ml, while the Dabok extract prolonged prothrombin time and activated partial thromboplastin time by 1.33 times and 2.21 times, respectively. All pepper extracts showed no erythrocyte hemolysis activity up to a concentration of 5 mg/ml. Our results suggest that native Korean peppers have the potential to serve as valuable sources of antioxidant, antidiabetic, and antithrombotic agents. This research also indicates the possibility of replacing improved pepper varieties, which incur significant seed usage fees, with native Korean pepper varieties.

Antioxidant Activities and Cytoprotective Effects of Lonicera japonica Thunb. Extract and Fraction against Oxidative Stress (인동덩굴 추출물과 분획물의 항산화 활성 및 산화적 스트레스에 대한 세포 보호 효과)

  • Lee, Ye Seul;Yun, Mid Eum;Lee, Yun Ju;Park, Young Min;Lee, Sang Lae;Park, Soo Nam
    • Microbiology and Biotechnology Letters
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    • v.46 no.1
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    • pp.18-28
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    • 2018
  • In this study, the antioxidant activities and cytoprotective effects against oxidative stress of Lonicera japonica Thunb. 50% ethanol extract and ethyl acetate fraction were investigated. Using the 1,1-diphenyl-2-picrylhydrazyl assay, the free radical scavenging activity (FSC50) of L. japonica Thunb. 50% ethanol extract and ethyl acetate fraction was determined as 152.00 and $77.25{\mu}g/ml$, respectively. To measure the reactive oxygen species (ROS) scavenging activity, the total antioxidant capacity (OSC50) was determined by using a luminol-dependent chemiluminescence assay. The antioxidant activity of the ethyl acetate fraction ($0.33{\mu}g/ml$) was approximately four times stronger than that of the 50% ethanol extract ($1.12{\mu}g/ml$). The protective effect against $^1O_2$-induced cellular damage of human erythrocytes (${\tau}_{50}$) was 46.0 min at $10{\mu}g/ml$ of the 50% ethanol extract and 52.3 min at $1{\mu}g/ml$ of the ethyl acetate fraction. We also investigated the cytoprotective effects against oxidative stress induced by $H_2O_2$ and the intracellular ROS scavenging activity in response to UVB irradiation and found that the extract and fraction protected human skin cells from damage and reduced ROS. These results confirmed that L. japonica Thunb. was a valuable plant-derived natural antioxidant with potential for development as an antioxidative functional ingredient.

Comparison of Antioxidant and ${\alpha}$-Glucosidase Inhibition Activities among Water Extracts and Sugar Immersion Extracts of Green Pepper, Purslane and Shiitake (청고추, 쇠비름, 표고버섯의 물 추출물 및 당침액의 항산화 활성 및 ${\alpha}$-Glucosidase Inhibition 활성 비교)

  • Lee, Sung Mee;Kang, Yun Hwan;Kim, Dae Jung;Kim, Kyoung Kon;Lim, Jun Gu;Kim, Tae Woo;Choe, Myeon
    • Journal of the East Asian Society of Dietary Life
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    • v.24 no.1
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    • pp.101-108
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    • 2014
  • This study was conducted in order to compare the biological activities of water extracts and sugar immersion extracts of green pepper (Capsicum annuum L.), purslane (Portulaca oleracea L.) and shiitake (Lentinula edodes (Berk.) Pegler) by measuring total polyphenol and flavonoid contents, antioxidant activities and inhibitory effects on ${\alpha}$-amylase and ${\alpha}$-glucosidase. The contents of total polyphenols and flavonoids were higher in water extracts than in sugar immersion extracts. The anti-oxidative activities of water and sugar immersion extracts were measured using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activity assay and reducing power assay. All extracts scavenged radicals in a concentration-dependent manner, and water extracts showed stronger radical scavenging activity and reducing power than sugar immersion extract. However, they all exhibited lower activities than ascorbic acid. Compared to the anti-diabetic drug acarbose, which was used as a positive control, the two types of extracts exhibited low ${\alpha}$-glucosidase inhibitory activities, although the activity of sugar immersion extracts were 2-fold higher than that of water extracts. ${\alpha}$-Amylase inhibitory action was not observed for any of the extracts. Finally, by cytotoxicity test, we confirmed that sugar immersion extracts were safer than water extracts. These results indicate that water extracts and sugar immersion extracts of green pepper, purslane and shiitake have different advantages in terms of their antioxidant and anti-diabetic effects, respectively.

Antioxidative, Antimicrobial, and Anti-proliferative Activities of the Floret and Stalk of Broccoli (Brassica oleracea L.) (브로콜리 꽃송이 및 줄기의 항산화, 항균 및 대장암 세포 생육억제효과)

  • Kim, Mi-Sun;Lee, Ye-Seul;Kwon, Ha-Young;Kim, Jong Sik;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.42 no.1
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    • pp.58-66
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    • 2014
  • In the course of study for a use for non-edible parts of broccoli (Brassica oleracea L), and the development of processed food utilizing these parts, edible floret and non-edible stalk were extracted with ethanol and different organic solvent fractions were prepared. With 10 different extracts and fractions, their useful components and various biological activities, such as antioxidant, antimicrobial and anti-proliferation activity, were investigated. The stalk has more abundant water soluble carbohydrate when compared with the floret, and floret has higher hexane-soluble pigments. Analysis of total flavonoid and total polyphenol contents showed that the floret has 1.5~1.99 times higher concentrations than the stalk. Among the fractions, ethylacetate (EA) fractions have the highest amount of total flavonoid and total polyphenol. The stalk and floret possessed 9.45 and 42.01 mg-total flavonoid/g, respectively. In the antioxidation activity assay, the EA fraction of floret showed strong radical scavenging activity and reducing power, while the n-hexane fraction of the stalk exhibited nitrite scavenging activity. In the antimicrobial activity assay, the EA fraction of floret showed a strong and broad-range of antibacterial activity, irrespective of gram positive or gram negative bacteria. In a while, the hexane and EA fractions revealed anti-proliferative effects against the human colorectal cancer cell HCT-116. Strong anti-proliferative activities were found in the hexane fraction of stalk (18.4% of cell viability), and the n-butanol fraction of floret (6.9% of cell viability). Our results suggest that the further study of the characterization of active fractions and the identification of active components from different parts of broccoli are needed to develop functional foods or novel plant-derived medicines.

A Comparison of the Components and Biological Activities in Raw and Boiled Red Beans (Phaseolus radiatus L.) (생팥과 증자팥의 성분 및 생리활성 비교)

  • Lee, Ryun Kyung;Kim, Mi-Sun;Lee, Ye-Seul;Lee, Man-Hyo;Lee, Jong Hwa;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.42 no.2
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    • pp.162-169
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    • 2014
  • In the course of study for the development of functional food using red beans (azuki beans, Phaseolus radiatus L.), the ethanol extracts from raw-red bean (RRB) and boiled-red bean (BRB) were prepared, and the components and various biological activities of both were compared. It was observed that the extraction yield, and the total polyphenol content, of the BRB were 1.2 times higher than that of the RRB. However, the contents of total flavonoid, total sugar and reducing sugar in the BRB were 30, 27.9 and 30.8% respectively when compared with those of RRB. In relation to antioxidative activity, both RRB and BRB exhibited moderate DPPH anion, ABTS cation, and nitrite scavenging activities and reducing power, though in all cases RRB demonstrated stronger activities than BRB. The extracts of RRB and BRB did not reveal any antimicrobial activities. In a ${\alpha}$-amylase inhibitory activity assay, RRB was higher than BRB, while BRB showed higher ${\alpha}$-glucosidase inhibitory activity than RRB. A strong and particular activity was observed in an anti-thrombosis activity assay of RRB. The extract of RRB demonstrated strong inhibitions against prothrombin and blood coagulation factors, with moderate thrombin inhibition. However, the extract of BRB did not exhibit any significant anti-thrombosis activity. Our results indicate that RRB has different, but useful biological activities, and loss or elimination of the biologically active substances in RRB occurs during the production of BRB. Therefore, to develop more functional foods from red beans, a study of suitable boiling, heating and drying processes is essential, and the efficient re-use of boiled waste water from the boiling process is necessary. These results could be applied to the further development of functional red bean beverages and sweat red bean pastes.

The protective effect of Eucommia ulmoides leaves on high glucose-induced oxidative stress in HT-29 intestinal epithelial cells (고당으로 유도된 산화적 스트레스에 대한 두충 잎 추출물의 장 상피 세포 보호 효과)

  • Han Su Lee;Jong Min Kim;Hyo Lim Lee;Min Ji Go;Ju Hui Kim;Hyun Ji Eo;Chul-Woo Kim;Ho Jin Heo
    • Food Science and Preservation
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    • v.31 no.1
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    • pp.183-196
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    • 2024
  • This study investigated the protective effect of the aqueous extract of Eucommia ulmoides leaves (AEEL) against high glucose-induced human colon epithelial HT-29 cells. The 2,2'-azino-bis (3-ethyl benzothiazoline-6-sulfonic acid) (ABTS), 1,1-diphenyl-2-picrylhydrazy (DPPH) radical scavenging activities, ferric reducing/antioxidant power (FRAP), and malondialdehyde (MDA) analyses indicated that AEEL had significant antioxidant activities. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed that AEEL increased cell viability against high glucose-, H2O2-, and lipopolysaccharide (LPS)-induced cytotoxicity in HT-29 cells. Also, the 2'-7'-dichlorodihydrofluorescein diacetate (DCF-DA) assay indicated that AEEL decreased intracellular reactive oxygen species (ROS) against high glucose-, H2O2-, and lipopolysaccharide (LPS)-induced cytotoxicity in HT-29 cells. AEEL showed inhibitory activities against α-glucosidase and inhibited the formation of advanced glycation end products (AGEs). AEEL showed significant positive effects on the viability and titratable acidity of L. brevis. The high-performance liquid chromatogram (HPLC) analysis identified chlorogenic acid and rutin as the major compounds of AEEL. These results suggested that AEEL has the potential to be used as a functional food source to suppress blood glucose levels and protect the gut from high glucose-induced oxidative stress.

Antioxidative Activity, Component Analysis, and Anti-elastase Effect of Aspalathus linearis Extract (루이보스 추출물의 항산화 활성, 성분 분석 및 엘라스테이즈 저해 효과)

  • Park, Soo-Nam;Yang, Hee-Jung;Won, Bo-Ryoung;Lim, Young-Jin;Yoon, Sun-Kyeong;Ji, Dong-Hwan;Choi, Jee-Yeon;Han, Seung-Joo;Lee, Chung-Woo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.33 no.4
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    • pp.251-262
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    • 2007
  • In this study, the antioxidative effects, inhibitory effects on elastase, and components of Aspalathus linearis extracts were investigated. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities ($FSC_{50}$) of extract/fractions of Aspalathus linearis were in the order: 50 % ethanol extract ($11.50\;{\mu}g/mL$) < deglycosylated flavonoid aglycone fraction ($8.47\;{\mu}g/mL$) < ethylacetate fraction ($4.76\;{\mu}g/mL$). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of some Aspalathus linearis extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activities were ethylacetate fraction ($OSC_{50},\;4.58\;{\mu}g/mL$) < deglycosylated flavonoid aglycone fraction ($2.20\;{\mu}g/mL$) < 50 % ethanol extract ($1.09\;{\mu}g/mL$). 50 % Ethanol extract showed the most prominent scavenging activity. The protective effects of extract/fractions of Aspalathus linearis on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Aspalathus linearis extracts suppressed photohemolysis in a concentration dependent manner, particularly 50 % ethanol extract exhibited the most prominent celluar protective effect (${\tau}_{50}$, 272.00 min at $50\;{\mu}g/mL$). Aglycone fractions obtained from the deglycosylation reaction of ethylacetate fraction among the Aspalathus linearis extracts, showed 3 bands in TLC and 3 peaks in HPLC experiments (360 nm). Three components were identified as luteolin (composition ratio, 18.24 %), quercetin (58.79), and kaempferol (22.97). TLC chromatogram of ethylacetate fraction of Aspalathus linearis extract revealed 7 bands and HPLC chromatogram showed 9 peaks, which were identified as isoorientin (composition ratio, 14.71 %), orientin (28.84 %), vitexin (5.63 %), rutin and isovitexin (12.73 %), hyperoside (9.24 %), isoquercitrin (5.40 %), luteolin (1.48 %), quercetin (17.61 %) and kaempferol (4.59 %) in the order of elution time. The inhibitory effect of aglycone fraction on elastase ($IC_{50},\;9.08\;{\mu}g/mL$) was very high. These results indicate that extract/fractions of Aspalathus linearis can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. And component analysis of Aspalathus linearis extract and inhibitory activity on elastase of the aglycone fraction could be applicable to new functional cosmetics for smoothing wrinkles.