• Title/Summary/Keyword: Solvent fractionation

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Isolation and Characterization of Helicobacter pylori Urease Inhibitor from Rubus coreanus Miquel (복분자(Rubus coreanus Miquel)로부터 Helicobacter pylori Urease Inhibitor의 분리 및 특성)

  • 양성우;호진녕;이유현;신동훈;홍범식;조홍연
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.5
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    • pp.769-777
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    • 2004
  • A Helicobacter pylori urease inhibitor from Rubus coreanus Miquel has been isolated and partially characterized for aiming to Prevent H. pylori growth and decrease harmful accumulation of ammonia in human gastric mucosa. We screened urease inhibitory activities in 519 extracts library prepared by solvent extraction from 173 kinds of edible plants, medicinal herbs, herbs and seaweeds using a colorimetric urease assay system. As results of primary and secondary screening, 70% acetone extract of Rubus coreanus Miquel was selected as potent candidate, showing about 24% inhibitory activity. The acetone extract was sequentially partitioned into RCE/RCWI and RCB/RCW2 layers with ethyl acetate and butanol. The major active component in RCW2, water layer from butanol fractionation was revealed to be peptidic or proteinous substance by inhibitory activity determination after pronase digestion and periodate oxidation. RCW2-IIIc a was isolated by sequential column chromatography on DEAE-Toyopearl 650C, Butrl-Toyopearl 650M and Sephadex LH-20. The isolated urease inhibitor RCW2-IIIc $\alpha$, was highly pure proteinous substance with molecular weight of 13kDa by high-performance gel permeation liquid chromatography. RCW2-IIIc$\alpha$ has about 5 times higher inhibitory activity than 70% acetone extract, showing high stability against heat treatment and peptic digestion.

Lipids from the rhizome of Cnidium officinalis Makino (천궁으로부터 lipid 의 분리 동정)

  • Kim, Hyoung-Geun;Jeon, Hyeong-Ju;Nguyen, Trong Nguyen;Lee, Dae Young;Baek, Nam-In
    • Journal of Applied Biological Chemistry
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    • v.64 no.4
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    • pp.343-349
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    • 2021
  • The rhizomes of Cnidium officinalis were extracted in aqueous MeOH, and the concentrate was fractionated via systematic solvent fractionation to EtOAc, n-BuOH, and aqueous fractions. The repeated column chromatography of EtOAc and n-BuOH fractions using silica gel, octadecyl silica gel, and Sephadex LH-20 as stationary phase to afford five lipids. They were identified to be methyl linoleate (1), linoleic aicd (2) 6-linoleoyl-𝛼-D-glucopyranosyl 𝛽-D-fructofuranoside (3), 1-linolenoyl-3-(𝛼-D-galactopyranosyl (1→6)-𝛽-D-galactopyranosyl) glycerol (4), and 1-linoleoyl-3-(𝛼-D-galactopyranosyl (1→6)-𝛽-D-galactopyranosyl) glycerol (5) on the basis of spectroscopic data such as IR, MS, and Nuclear magnetic resonance (NMR). Compounds 1 and 3-5 were isolated for the first time from this plant in this study. The NMR data of fatty acids 1 and 2 reported in literatures are different each other. Authors identified the NMR data without ambiguity. Compound 3, a conjugate of sucrose and fatty acid, and compounds 4 and 5, digalactosyl monoglyceride, are very rarely occurred in natural source. Through the immune enhancement and anticancer activity of the reported lipid compounds, the potential as various pharmacologically active materials of Cnidium officinalis rhizome can be expected.

Anti-inflammatory effects of Chamaecyparis obtusa (Siebold & Zucc.) Endl. Leaf Fractions (편백(Chamaecyparis obtusa (Siebold & Zucc.) Endl.) 잎 분획물의 항염증 효과)

  • Yong-Jin Kwon
    • Journal of the Korean Applied Science and Technology
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    • v.40 no.6
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    • pp.1268-1277
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    • 2023
  • In this study, to evaluate the possibility of utilizing Chamaecyparis obtusa (Siebold & Zucc.) Endl. (C. obtusa) leaf fractions as anti-inflammatory functional materials, C. obtusa extract extracted with 99% ethanol (CO99EL) was fractionated with hexane (CO99EL-H), chloroform (CO99EL-C), ethyl acetate (CO99EL-E), butanol (CO99EL-B) and distilled water (CO99EL-W). The anti-inflammatory effects of each fraction was performed using lipopolysaccharide (LPS)-induced RAW264.7 mouse macrophages. Cytotoxicity was highest in CO99EL-H and CO99EL-C and lowest in CO99EL-W. Interestingly, LPS-induced iNOS expression and NO production were significantly reduced by CO99EL-H and CO99EL-E, and COX-2 expression was significantly reduced by CO99EL-B and CO99EL-W. In addition, interleukin (IL)-1𝛽, an inflammatory cytokine increased by LPS, was significantly reduced by CO99EL-C, CO99EL-E, CO99EL-B and CO99EL-W, and IL-6 was significantly reduced by CO99EL-B and CO99EL-W. Therefore, the janus kinase (JAK)/signaling transducer and activator of transcription (STAT) signaling pathway activated by LPS was significantly reduced by CO99EL-H and CO99EL-C, and the mitogen-activated protein kinase (MAPK) signaling pathway was slightly reduced by CO99EL-H and CO99EL-C. However, nuclear factor (NF)-𝜅B activity was not reduced by any fractions. Based on the results of this study, it was confirmed that CO99EL fractions have different anti-inflammatory mechanisms depending on the solvent used for fractionation.

Biological activity and analysis of α-glucosidase inhibitor from mulberry (Morus alba L.) wine (오디와인의 생리활성 및 α-glucosidase 저해제의 분석)

  • Son, Woo-Rim;Choi, Sang-Won
    • Food Science and Preservation
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    • v.20 no.6
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    • pp.877-885
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    • 2013
  • Wine extracts of four different berry fruits, such as mulberry, blueberry, strawberry, and raspberry, were investigated for antioxidant, anti-tyrosinase, and ${\alpha}$-glucosidase activities by using in vitro assays. Additionally, quantitative changes of ${\alpha}$-glucosidase inhibitor in mulberry wine were determined by HPLC according to mulberry cultivars and fermentation process. Among four berry wines examined, mulberry wine showed the most potent ${\alpha}$-glucosidase inhibitory activity with 69.37% at 0.23 mg/mL, while blueberry and strawberry wines exhibited the strongest inhibition against DPPH radical and tyrosinase activity, respectively. Four compounds were isolated and purified from mulberry wine by a series of isolation procedures, such as solvent fractionation, and Diaion HP-20, ODS-A, and Sephadex LH-20 column chromatographies. Among them, Comp. 4 exerted the strongest ${\alpha}$-glucosidase inhibitory activity ($I_C_{50}=31.57{\mu}M$), and its chemical structure was identified as quercetin by UV and NMR spectral analysis. Finally, the "Daeseongppong" (16.83 ppm) muberry wine had larger amount of quercetin than the "Iksuppong" (14.85 ppm) and "Cheongilppong" (8.92 ppm) mulberry wines, but their contents of three mulberry wines decreased considerably with aging process. These results suggest that mulberry wine containing quercetin acted as ${\alpha}$-glucosidase inhibitor may be useful as a potential functional wine for improving diabetic disorder.

Pro-apoptotic and Anti-adipogenic Effects of Proso Millet (Panicum miliaceum) Grains on 3T3-L1 Preadipocytes (기장(Panicum miliaceum)의 마우스 3T3-L1 세포에 대한 에폽토시스 유발 및 지방세포형성 억제 효능)

  • Jun, Do Youn;Lee, Ji Young;Han, Cho Rong;Kim, Kwan-Pil;Seo, Myung Chul;Nam, Min Hee;Kim, Young Ho
    • Journal of Life Science
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    • v.24 no.5
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    • pp.505-514
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    • 2014
  • To examine the anti-obese activity of miscellaneous cereal grains, 80% ethanol extracts from eight selected miscellaneous cereal grains were compared for their cytotoxic effects on 3T3-L1 murine preadipocytes. The ethanol extract of proso millet exhibited the highest cytotoxicity. Further fractionation of the ethanol extract with methylene chloride, ethyl acetate, and n-butanol showed that the cytotoxicity of the ethanol extract was mainly partitioned into the butanol fraction. As compared with differentiated mature adipocytes, 3T3-L1 preadipocytes were more susceptible to the cyctotoxicity of the butanol fraction. When each organic solvent fraction (25 ${\mu}g/ml$) was added during the differentiation period for 6 days, the cell viability was not affected significantly except for the butanol fraction, but the intracellular lipid accumulation declined to a level of 81.5%~50.3% of the control. The Oil Red O staining data also demonstrated that the ethanol extract as well as the butanol fraction could inhibit the differentiation of 3T3-L1 preadipocytes into mature adipocytes. The presence of the butanol extract during the induced adipocytic differentiation also resulted in a significant reduction in the expression levels of critical adipogenesis mediators $(C/EBP{\alpha}$, $PPAR{\gamma}$, aP2, and LPL) to a barely detectable or undetectable level and the cells retained the fibroblast-like morphology of 3T3-L1. In 3T3-L1 cells, the cytotoxicity of the butanol fraction (50-100 ${\mu}g/ml$) was accompanied by mitochondrial membrane potential (${\Delta}{\psi}m$) loss, caspase-3 activation, and PARP degradation. Taken together, these results indicate that proso millet grains possess pro-apoptotic and anti-adipocytic activities toward adipocytes, which can be applicable to prevention of obesity.

Anticarcinogenic Effects of Extracts from Gloiopeltis tenax (참가사리 분획물의 암 예방효과)

  • Jung, Young-Hwa;Jung, Bok-Mi;Shin, Mi-Ok;Bae, Song-Ja
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.4
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    • pp.395-401
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    • 2006
  • In this study, we investigated anticarcinogenic effects of extracts from Gloiopeltis tenax (GT). GT was extracted with methanol (GTM), which was then further fractionated into four fractions by using solvent fractionation method, affording methanol (GTMM), hexane (GTMH), butanol (GTMB) and aqueous (GTMA) soluble fractions. We determined the cytotoxic effects of these fractions on cancer cells by MTT assay. Among various fractions of GT, the GTMM showed the strongest cytotoxic effect at concentration of $150{\mu}g/mL$, displaying 95.97% on HepG2 cell lines and 93.64% on HT-29 cell lines, respectively. And, the anti-proliferative effect of GT was accompanied by a marked in increase of levels of Bad, Bax, Bok and Bak protein and activation of caspase-3, caspase-7 and PARP protein. Also, we observed quinone reductase (QR) induced effects in all fraction layers of GT on HepG2 cells. The QR induced effects of the GTMM and GTMB on HepG2 cells at concentration of $60{\mu}g/mL$ showing inductive indexes of 2.86 and 2.04 compared to the control value of 1.0.

Enzymological Evaluation of Oral Inflammation inhibitory activity by Aloe vera peel extract (Aloe vera peel 추출물에 의한 구강염증 저해 효과의 효소학적 평가)

  • Pack, Chung-Sun;Ryu, Il-Hwan;Lee, Kap-Sang
    • Korean Journal of Food Science and Technology
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    • v.33 no.6
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    • pp.753-759
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    • 2001
  • To evaluate antiinflammation of Aloe vera peel, antiimflammation substances were extracted from Aloe vera peel and identified, and we investigated the effect of the its substance the inhibitory effect on the activity of hyaluoronidase, elastase, collagenase and prostaglandin endoperoxide synthase. The water extract from Aloe vera peel were successfully purified with solvent fractionation, silica gel column chromatography, preparative thin layer chromatography and UV spectrometer. Two purified active substances were identified as aloe-emodin and barbaloin by Mass Spectrometer, $^1H-NMR$ and FT-IR. Aloe-emodin and barbaloin. $IC_{50}$ values of aloe-emodin and barbaloin against hyaluronidase activity were 40 and $70\;{\mu}g/mL$, respectively. Leuckocyte elastase, which is related to the destruction of various tissue, $IC_{50}$ values of them were 50 and $60\;{\mu}g/mL$, respectively. $IC_{50}$ values of aloe-emodin and barbaloin against collagenase activity were 40 and $60\;{\mu}g/mL$, respectively. and $IC_{50}$ values of aloe-emodin and barbaloin aganist the prostaglandin endoperoxide synthase, which play an important role in inflammatory reactions, were 40 and $70\;{\mu}g/mL$, respectively. Inhibitory effects of aloe-emodin, barbaloin and aspirin against carrageenan paw edema were 74.9, 52.9 and 51.9% as inhibiton percentage, respectively, at dose of 100 mg/kg and that of indomethancin was 49.7 at dose of 10 mg/kg. Cell cytotoxicity of barbaloin against human gingival cells was lower than that of aloe-emodin. Aloe-emodin and barbaloin did not show cytotoxicity against human gingival cells at concentration of 1.0 and $5.0\;{\mu}g/mL$, However, aloe-emodin and barbaloin showed less cytotoxicity than chlorhexidine, which usually have been used as the agent of anticaries and antiinflammation. These results suggested that aloe-emodin and barbaloin from Aloe vera peel have the effect of anticaries and antiinflammation.

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Evaluation for the Antimicrobial, Antioxidant and Anti-thrombosis Activity of Korean Traditional Liquors (시판 전통주의 항균, 항산화 및 항혈전 활성 평가)

  • Ryu, Hee-Young;Kum, Eun-Joo;Bae, Kyung-Hwa;Kim, Young-Kwan;Kwun, In-Sook;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.35 no.3
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    • pp.238-244
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    • 2007
  • To identify the useful biological activities and the superiority in quality of Korean traditional liquors, we prepared the alcohol-free concentrates from different types of commercial traditional liquors, Takju ($T1{\sim}T3$), Yakju/Cheongju ($Y-1{\sim}Y-5$), Fruit wine (F-1) and Soju (S-1, S-2), respectively. We investigate their compositions as well as antimicrobial, antioxidant, and anti-thrombosis activity. Among the 11 traditional liquors, Y-3, Y-4, Y-5 and F-1 showed higher total-polyphenol, total-flavonoids and reducing sugars than the others. The strong antibacterial and anti-thrombosis activities were identified in Y-3, Y-4, Y-5 and F-1, and a minor antioxidant activity was found in F-l. The antibacterial activity of the Y-3, Y-4, Y-5 and F-1 alcohol-free concentrates showed a broad-spectrum, and growth inhibition was found in gram-positive, gram-negative, and ampicillin-resistant bacteria. The sequential solvent fractionation of Y-3, Y-4, Y-5 and F-1, and following analysis showed that ethyl acetate fractions of Y-3, Y-4, Y-5 and F-1 possess strong antibacterial and anti-thrombosis activity. Especially, the ethyl acetate fractions of Y-3, Y-4 and F-1 showed superior anti-thrombosis activity compared than that of aspirin. Our results suggest that the useful substances are produced from substrates and edible plant added during the fermentation, and the Korean traditional liquors could be developed as strong antibacterial and anti-thrombosis agents.

Studies on the Development of Natural Preservatives from Natural Products (전통식품 및 천연물에서 천연보존료 개발에 관한 연구)

  • Kim, Hee-Yun;Lee, Young-Ja;Hong, Ki-Hyoung;Kwon, Yong-Kwan;Lee, Ju-Yeun;Kim, So-Hee;Ha, Sang-Chunl;Cho, Hong-Yon;Chang, Ih-Seop;Lee, Chul-Won;Kim, Kil-Saeng
    • Korean Journal of Food Science and Technology
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    • v.31 no.6
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    • pp.1667-1678
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    • 1999
  • Certain parts of 190 kinds of medicinal herbs and 171 kinds of original materials of food were extracted by methanol. The extracts were tested their microbial inhibition activities against several food spoilage microorganisms, Micrococcus luteus, Bacillus subtilis, Bacillus cereus, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Escherichia coli, Saccharomyces cerevisiae, Candida albicans, Penicillium citrinum, Aspergillus flavus and Aspergillus niger. The methanol extracts of Cornus officinalis, Evodia officinalis, Glycyrrhiza glabra, Salvia miltiorrhiza. Schizandrae fructus, Coptidis rhizoma, aroma hop and bitter hop were shown inhibitory effect on certain species of gram(+) bacteria. Aroma hop and bitter hop were shown inhibitory effect on certain species of gram(-) bacteria. The methanol extract of Salvia miltiorrhiza exhibited a strong antibacterial activities. It was purified by solvent fractionation, silicagel column chromatography, prep. TLC, prep. HPLC. The purified active substance was identified as cryptotanshinone by EIMS, $1^H-NMR,\;{13}^C-NMR$ and DEPT. Cryptotanshinone showed a strong antibacterial activity against gram positive bacteria $(MIC\;:\;3.91{\sim}62.50\;{\mu}g/mL)$. Especially, this compound was the most strong activity against Bacillus subtilis $(MIC\;:\;3.91\;{\mu}g/mL)$.

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