• 제목/요약/키워드: TBA assay

검색결과 47건 처리시간 0.032초

작약 약침액이 tert-butyl hydroperoxide 로 유도된 흰쥐 배양 간세포의 지질과산화반응 및 항산화효소 활성에 미치는 영향 (Effects of Paeoniae Radix Aqua-Acupuncture Solution on Tert-Butyl Hydroperoxide Induced Lipid Peroxidation and Antioxidative Enzymes in Cultured Rat Liver Cells)

  • 문진영
    • Journal of Acupuncture Research
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    • 제17권3호
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    • pp.176-187
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    • 2000
  • Objectives : This study was purposed to investigate the antioxidative effects of Paeoniae radix aqua-acupuncture solution(PR) on culture liver cell system, lipid peroxidation and antioxidative enzyme activities in tert-butyl hydroperoxide(t-BHP) treatmented conditions. Methods : Cultured normal rat liver cell(Ac2F) were prepared and incubated with or without PR(at 2% volume in culture medium). After 16~18hr, cells placed in DMEM medium without serum, and then incubated with 1mM t-BHP for 2hr. Viable cells were detected by MTT assay, and the levels of lipid peroxide(LPO) were measured by TBA method. And catalase activity was measured as the decrease in hydrogen peroxide absorbance at 240nm on spectrophotometer using 30mM hydrogen peroxide. Superoxide dismutase(SOD) were assayed by recording the inhibition of nitro blue tetrazolium reduction with xanthine and xanthine oxidase. Glutathione peroxidase(GPX) activity was determined by the modified coupled assay developed by Paglia and Lawrence. The reaction was started by addition of 2.2mM hydrogen peroxide as substrate. The change in absorbance at 340nm was measured for 1min on spectrophotometer. Glutathione-S-transferase(GST) activity was assayed with CDNB as substrate and enzyme activity of GST towards the glutathione conjugation of CDNB. Results : Cell killing was significantly enhanced by addition of t-BHP compared to those of untreated group. PR pretreated cell resisted the toxic effects of t-BHP. LPO levels of t-BHP treatment group were significantly higher than other groups. This increased level was significandy reduced by PR pretreatment. The t-BHP treatment resulted in a decrease of catalase, GPX and GST activities. By contrast, PR pretreatment markedly increased compare to those of untreated groups. Conclusions : T-BHP which can produce intracellular free radical was used for inducer of the peroxidation of cellular lipids. PR protected the cell death induced by t-BHP and significantly increased cell viabiliry in the normal rat liver cell, and showed effective inhibition of lipid peroxidation, and elevations of catalase, GPX and GST activities. These results suggested that PR might play a protective role in lipid peroxidation by free radicals.

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Inhibitory effects of medicinal plants on elastase activity and biological activities in the active Plant extracts

  • 이건국;조중재;최중덕
    • 대한화장품학회지
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    • 제23권2호
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    • pp.7-22
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    • 1997
  • Inhibitory effects of 200 medicinal plants on elastase activity were screened. Among the 200 plants, six plant extracts exhibited more than 65% of inhibition on elastase activity by their total methanol extracts at 1, 000$\mu\textrm{g}$/$m\ell$ as a final concentration. For six active plants including Areca catechu, Cinnamonum cassia, Myristica frograns, Curcumma longa, Alponia katsumadai, and Dryopteris crassirrhizoma, the inhibitory effects of their comsituents were examined on the activity of human leukocyte elastase, hyaluronidase and lipid peroxidation. In lipid peroxidation assay using TBA method, three of the 6 plants including Curcumma longa, Areca Catechu and Alponia katsumadai exhibited more than 70% of inhibition at the concentration of 1, 000$\mu\textrm{g}$/$m\ell$, but only one plant, Areca Ctechu showed high inhibitory effect on hyaluronidase activity. The results suggest that medicinal plants showing several biological activities may be potent inhibitors of anti-aging process on skin and that might be useful for application in cosmetics.

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Antioxidant Activities of Isoflavones from the Rhizomes of Belamcanda chinensis on Carbon Tetrachloride-Induced Hepatic Injury in Rats

  • Jung, Sang-Hoon;Lee, Yeon-Sil;Lim, Soon-Sung;Lee, Sanghyun;Shin, Kuk-Hyun;Kim, Yeong-Shik
    • Archives of Pharmacal Research
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    • 제27권2호
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    • pp.184-188
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    • 2004
  • The present study was carried out to clarify whether tectorigenin and tectoridin isolated from the rhizomes of Belamcanda chinensis (Iridaceae) inhibit hepatic damage induced by carbon tetrachloride ($CCl_4$)-intoxication in rats by the experimental methods in vitro and in vivo. Tectorigenin and tectoridin exhibited a significant decrease in serum transaminase activities elevated by hepatic damage induced by $CCl_4$-intoxication in rats, as well as in a lipid peroxidation causing a significant decrease in malondialdehyde (MDA) production by thiobarbituric acid (TBA)-reactant assay. Both compounds also showed strong increase in the antioxidant enzymes such as hepatic cytosolic superoxide dismutase (SOD), catalase and glutathione peroxidase (GSH-px) activities in $CCl_4$-intoxicated rats. These results suggested that tectorigenin and tectoridin isolated from the rhizomes of B. chinensis possess not only the antioxidative, but also the hepatoprotective activities in $CCl_4$ -intoxicated rats.

Antioxidant Activity of Salad Vegetables Grown in Korea

  • Xin Zao;Song, Kyung-Bin;Kim, Mee-Ree
    • Preventive Nutrition and Food Science
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    • 제9권4호
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    • pp.289-294
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    • 2004
  • The antioxidant activity of forty two kinds of salad vegetables grown in Korea was evaluated. Methanol extract of freeze-dried vegetable was assayed by radical scavenging activity using 1,2-diphenyl-2-picrylhydrazyl (DPPH) and Fe^{2+}$-catalyzed lipid peroxidation inhibition by TBA method. Total phenolics were determined colorimetrically using Folin-Ciocalteu reagent. The highest radical scavenging activity was expressed by perilla leaf, followed by dandelion leaf, red and green leafy lettuce, of which $IC_{50}$ was less than 0.10 mg/mL. Angelica leaf showed the highest inhibitory action for lipid peroxidation with $95\%$, and then dandelion leaf, water spinach, and perilla leaf inhibited over $80\%$. However, lettuce (Iceberg) and young Chinese cabbage exhibited the lowest antioxidant activity based on both assay methods. Highly positive correlations between antioxidative activities and total phenolics were observed (p < 0.001). The results suggested that salad vegetables, especially perilla leaf, leafy lettuce, dandelion or angelica, could be used for easily accessible sources of natural antioxidants.

어류의 가열조리 및 보존에 의해 생성되는 변이원성 물질의 정량적 해석과 제어법 (Screening and prevention of the mutagenicity for fishes accordind to cookery and storage)

  • 홍이진;이준경;구성자
    • 한국식품조리과학회지
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    • 제16권6호
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    • pp.652-662
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    • 2000
  • 섭취 빈도가 높은 어류중 붉은 살 생선인 고등어(Mackerel), 삼치(Saury pike), 방어(Yellowtail), 연어(Salmon), 흰 살 생선인 조기 및 염장조기(Croaker, Salted Croaker), 및 갈치(Trichiurus)를 시료로 하여 , 가스그릴, 전기그릴과 microwave oven으로 가열 조리한 후 일정기간 동안 냉장 및 냉동 보존한 시료와 보존한 시료를 microwave로 재가열하였을 때 변이원성을 in vitro test로 Ames/salmonella assay를, in vivo test 로 Micronucleous test를 실시하여 검색하고, 그 제어방법으로 항암성분을 많이 함유하고 있는 녹황색 채소즙으로 전처리하여 돌연변이 억제능을 검토하고자 한다. 또한, 지질의 과산화도를 측정하여 변이원 생성과 과산화 지질간의 상관성을 검토한 결과는 다음과 같다. 1.어류의 가열조리에 의해 대조군(생시료)보다 변이원성이 유의적으로 증가되었는데 , 생성된 변이원은 모두 농도 의존성을 나타내었으며, 5 mg/plate인 농도구간에서 돌연변이원성이 유의적으로 높게 나타났다(p<0.05). 2. 조리온도가 높아질수록 변이원성이 유의적으로 높아졌으며 대사활성물질을 첨가했을 경우 무첨가시보다 2-4 배정도 돌연변이원성이 높게 나타났다. 3. 조리기기에 따른 복귀 돌연변이 균수는 가스그릴과 전기그릴군 간에는 유의적인 차이가 없었으나 MW에서 조리한 시료의 경우에는 유의적으로 낮게 나타났다(p<0.05). 4. 생선 종류별 변이원성을 검색한 결과 흰살 생선이 붉은 살 생선에 비해 유의적으로 낮은 변이원성을 나타내었으며, 붉은 살 생선간에 유의적인 차이는 나타나지 않았으나 고등어가 약간 높은 변이원성을 나타내었고, 흰살 생선중에서도 유의차는 없었으나 염장조기가 가장 낮은 변이원성을 나타내었다. 5. 채소즙 및 녹차 추출물로 처리 후 가열조리을 한 시료의 경우 모두 대조군에 비해 유의적으로 변이원성이 억제되었으며, 녹차>생강>무순으로 변이원성 억제효과가 나타났다. 6. 저장에 의해 돌연변이원성이 유의적으로 증가하였는데, 냉장 저장시 저장 3일째부터, 냉동저장시에는 저장 7일째부터 변이원성이 유의적으로 증가하였다. 냉동저장보다 냉장저장시 더 높은 변이원성을 나타내었고 MW에 의한 재가열에 의해 변이원성이 증가되었다. 7. Micronucleus test 결과, 조리온도가 증가할수록 골수 소핵 형성이 유의적으로 증가되었고, 채소즙 처리군의경우 생강>녹차>무순으로 골수 소핵 형성 억제효과를 보였다. 8. 조리, 재가열 및 저장 조건에 따른 산패도(TBA value)를 측정한 결과 저장기간이 길어질수록 산패도가 증가하였고 냉장 저장시료의 경우 저장 3일째부터 유의 적으로 산패도가 증가되었으며 냉동 저장 시료의 경우에 는 저장 7일째부터 유의적으로 높은 산패도를 나타내었다. 냉장저장 시료가 냉동저장 시료보다 더 높은 산패도를 나타내었으며, microwave에 의한 재가열에 의해 산패도가 증가되었다. 9. 채소즙 및 녹차 추출물로 처리 한 고등어의 DPPH에 대한 전자공여능(%EDA)측정 결과는 양파>녹차>생강>양배추 순으로 양파의 전자공여능이 가장 높게 나타났다. 10.전기그릴 26$0^{\circ}C$에서 조리한 시료의 냉장저장 및 냉동저장에 따른 변이원성과 과산화지질 생성간의 상관관계는 냉장 저장했을 때 0.93으로 높은 상관관계를 나타내었고 재가열했을 경우 0.98로 상관관계가 더 높게 나타났다. 냉동저장 역시 0.92를 나타내어 상당히 높은 상관관계를 나타내었으나 재가열했을 경우에는 0.89로 오히려 낮은 상관관계를 보였다.

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왕쥐똥나무잎 추출물의 항산화 활성 (Antioxidant Activities of Extracts from Ligustrum ovalifolium H. Leaves)

  • 김연숙;이승재;황진우;김이화;박표잠;정재현
    • 한국식품영양과학회지
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    • 제40권12호
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    • pp.1642-1647
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    • 2011
  • 본 연구는 왕쥐똥나무잎(Ligustrum ovalifolium H.) 추출물의 항산화 활성을 탐색하고자 물과 에탄올로 각각 추출하였다. 왕쥐똥나무잎에 포함된 총 폴리페놀과 총 플라보노이드 함량을 측정한 결과, 물 추출물($105.5{\pm}1.31$ mg GAE/g extract)과 에탄올 추출물($102.1{\pm}1.82$ mg GAE/g extract)은 비슷한 총 폴리페놀 함량을 함유하고 있었으며, 총 플라보노이드 함량 또한 물 추출물($84{\pm}1.72$ mg CE/g extract)과 에탄올 추출물($82.8{\pm}1.65$ mg CE/g extract)이 비슷한 양을 함유하고 있었다. ESR을 이용한 라디칼 소거능을 측정한 결과, hydroxyl 라디칼을 제외한 DPPH, alkyl 라디칼 소거능은 물 추출물 및 에탄올 추출물 모두 높은 활성을 나타내었다. 특히 alkyl 라디칼 소거능은 비타민 C와 같은 활성을 나타내었다. ABTS를 이용한 라디칼 소거활성, FRAP을 이용한 총항산화능 측정 및 환원력을 통한 항산화 활성을 측정한 결과에서도 왕쥐똥나무잎 추출물이 뛰어난 항산화효과를 가지고 있음이 확인되었다. 특히, FTC 및 TBA법을 이용한 지질과산화 억제 효능을 살펴본 결과, 왕쥐똥나무잎 물추출물과 에탄올 추출물 모두 지질과산화 억제효과가 높았으며 ${\alpha}$-토코페롤보다 활성이 훨씬 우수하였다. 한편, 세포독성을 살펴보기 위하여 정상 간세포(human liver, Chang cells)를 이용하여 MTT assay를 통한 세포의 생존율을 살펴본 결과 물 추출물 및 에탄올 추출물 모두 0.5 mg/mL의 농도까지 전혀 독성을 나타내지 않았다. 따라서 이 연구를 바탕으로 앞으로 왕쥐똥나무잎을 이용한 천연 항산화제로서의 가능성에 대한 추가적인 연구가 필요할 것으로 판단된다.

방사선 조사된 고불포화지방산 식이를 섭취한 Mouse의 산화적 스트레스 (Oxidative Stress of Mouse Fed Irradiated Diet Containing High Unsaturated Fatty Acid)

  • 서대영;박선영;강명희;서광선;이선영
    • Journal of Nutrition and Health
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    • 제39권7호
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    • pp.599-609
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    • 2006
  • In this study, we investigated the in vivo effect by intake of the irradiated foods such as mackerel and sesame seed which are high in unsaturated fatty acid through TBARS (thiobarbituric acid reactive subtance) and the tissue pathological and genotoxicological test. Thirty two ICR mice are divided into four groups, one non-irradiated (control) group and three irradiated (5, 10, 20 kGy, respectively) groups. Sesame seed and pulverized mackerel in modified AIN93M diet were mixed together then divided into four identical parts. Three parts of them were irradiated by doses of 5, 10, and 20 kGy. These experimental diet were fed to each group for 4, 8 and] 6 weeks. The results of the study were as follows: No significant differences in weight gain were found in each group. Peroxide value of the irradiated diet was higher than that of the non-radiated one and also increased according to the storage period. TBA values in plasma, liver, kidney and Peyer's patch were not significantly different among 4 groups. DNA% in tail, tail length (TL) and tail moment (TM) values of the blood lymphocyte in 4, 8 and 16 week groups and the liver in 16 week group were much measured over the control DNA % in tail of kidney of 8 week group was significantly than the control and TL and TM of 8 week and TM of 16 week groups showed a tendency of higher values. By Peyer's DNA % in tail of 8 week group, DNA % in tail and TM of 16 week groups increased significantly over the control, Ultrastructural examination shows myeline figures and swollen mitochondria in parietal cells and intestinal epithelial cells of 8 and 16 weeks groups. After this study, we need further investigations on the safety of highly consumed foods which contain high contents of unsaturated fatty acid, largely imported and which are possible to be irradiated.

보라우무 (Symphyocladia latiuscula) 추출물의 항산화활성 (The Antioxidant Activity in Extracts of Symphyocladia latiuscula)

  • 박혜진;정해영;최재수
    • 한국수산과학회지
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    • 제31권6호
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    • pp.927-932
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    • 1998
  • 일본, 따렌 등에 분포되어 있고 우리나라에서 동해안, 서해안, 남해에서 흔히 볼 수 있는 해조류로서, 조간대의 바위위에서 서식하는 빨강 검둥이과 홍조류인 보라우무 (Symphyocladia latiuscula)를 대상으로 하여 안정한 free radical인 1,1-diphenyl-2-picrylhydrazyl (DPPH)를 사용하여 그 소거능을 측정하고, 또한 생체내의 과산화지질을 평가하는 방법으로 과산화지질 분해산물인 malondialdehyde를 thiobarbituric acid(TBA)로 비색정량하는 방법과 Hepatocyte에 의해 생성되는 free radical 생성량 측정 등의 다양한 방법을 통하여 항산화능을 측정하였다. DPPH radical을 이용한 항산화활성 검색에서 강력한 radical 소거 효과를 나타내었다. 각 용매 분획별로 항산화활성을 살펴본 결과 수층을 제외한 모든 분획들에서 농도 의존적으로 항산화활성이 나타났다. 특히 dichloromethane과 hexane 가용성 분획의 항산화활성이 가장 크게 나타났다. 지질과산화 억제 효과에서도 유사한 결과를 보였으며 dichloromethane 가용성 분획의 항산화활성이 가장 높게 나타났으며 L-ascorbic acid의 항산화활성과 비교하여 더 높은 활성을 나타내었다. Free radical 생성 억제 효과에서도 dichloromethane 가용성 분획의 경우 control과 비교하여 0.1 mg/ml에서 $50\%$ 이상의 저해효과를 나타내었다.

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Effect of Feeding Cyanidin 3-glucoside (C3G) High Black Rice Bran on Nutrient Digestibility, Blood Measurements, Growth Performance and Pork Quality of Pigs

  • Kil, D.Y.;Ryu, S.N.;Piao, L.G.;Kong, C.S.;Han, S.J.;Kim, Y.Y
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권12호
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    • pp.1790-1798
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    • 2006
  • Two experiments were conducted to investigate the effect of feeding cyanidin 3-glucoside (C3G) high black rice bran on nutrient digestibility, blood measurements, growth performance and pork quality of pigs. In Exp. I, a total of fifteen pigs (19.91${\pm}$1.80 kg, average initial body weight) were used in assay of nutrient digestibility and blood measurements. All pigs were allotted to 5 treatments with 3 replicates according to a completely randomized design (CRD) in an individual metabolic crate. Treatments included 1) CON: basal diet, 2) BRB-2: basal+brown rice bran 2%, 3) BRB-4: basal+brown rice bran 4%, 4) CRB-2: basal+C3G high black rice bran 2% and 5) CRB-4: basal+C3G high black rice bran 4%. The digestibility of dry matter (DM), crude protein (CP), crude fat (CF), crude ash (CA) and crude fiber (CF) was not affected by dietary treatments. Serum triglyceride (TG) and high density lipoprotein (HDL) cholesterol concentrations were not affected by addition of C3G high black rice bran. However, at the end of experiment, pigs fed rice bran showed decreased tendency in total cholesterol concentration. Especially pigs fed C3G high black rice bran showed significantly lower total cholesterol concentration compared to pigs fed brown rice bran (p<0.03). There was numerically lower total cholesterol concentration with increasing levels of black rice bran in the diet. In terms of serum glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT), there were no significant differences among treatments, even though pigs fed CRB-4 showed the lowest GOT concentration compared to other pigs. In Exp. II, sixteen finishing pigs (average initial body weight 89.96${\pm}$0.35 kg) were divided into 4 treatments to investigate the effect of feeding C3G high black rice bran on growth performance and pork quality. There were no significant differences in average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR) among the treatments. Pigs fed C3G high black rice bran showed numerical decrease in ADG and increase in FCR while not effecting feed intake. There was no significant difference in live weight, carcass weight, carcass rate, backfat thickness and carcass grade. However, pigs fed C3G high black rice bran tended to show lower backfat thickness than pigs fed basal diet. Pigs fed C3G high black rice bran showed a tendency of decreased TBA value than pigs fed basal diet, although there was no overall significant difference among treatments. In conclusion, nutrient digestibility, blood measurements, growth performance and pork quality were not significantly affected by feeding C3G high black rice bran to pigs. However, C3G high black rice bran might have an effect on lowering serum total cholesterol and decrease the TBA value in pork compared to control group and these effects might be due to high concentration of antioxidative compounds in C3G high black rice bran.

Lactobacillus spp.와 Bacillus coagulans의 Glutathione Sulphydryl 함유율과 황산화 활성 (Glutathione Sulphydryl Contents and Antioxidant Activities of Lactobacillus spp. and Bacillus coagulans)

  • 변정열;윤영호
    • Journal of Dairy Science and Biotechnology
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    • 제22권2호
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    • pp.91-97
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    • 2004
  • 활성산소의 세포막 파괴 효과로부터 보호작용과 항산화 활성이 강한 균주를 선별하기 위하여 33균주에 대한 세포네 glutathione 함유율과 linoleic acid 산화억제활성을 측정하였다. Lactobacillus casei CU4114 25.15 umol/g으로 가장 높은 함유율을 보이고 있으며Lactobacillus rhamnosus CU4201, Lactobacillus plantarum CU4203이 14.0umol/g으로 통계적인 유의성을 나타내는 수준으로 높은 함유율을 나타내었다. 포자형 성균주중의 glutathione 함유율 측정 결과 Lactobacillus sporogenes CU4109, Sporolactobacillus inulinus CU4110 은 Bacillus coagulans 균주의 GSH 함유율보다 유의성이 인정되는 높은 함유율을 나타내었다. L. acidophilus CU4111, L. casei CU4114, L. brevis CU4206, L. helveticus Cu4231은 Lactobacillus spp. 중 높은 항산화 활성을 나타내었고 Bacillus coagulans의 항산화 활성범위는 6.02${\sim}$51.72%로 나타났으며 Bacillus coagulans 균주의 항산화 활성이 Lactobacillus spp.의 산화억제활성보다 높은 성향을 나타내었다.

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