• Title/Summary/Keyword: silica-gel column chromatography

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Isolation and Structural Determination of Anti-Helicobacter pylori Compound from Fungus 60686. (곰팡이에서 분리한 Helicobacter pylori 항균물질의 분리 및 구조규명)

  • 남궁준;연승우;백남수;김태한;김영호;김창진;김기원
    • Microbiology and Biotechnology Letters
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    • v.26 no.2
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    • pp.137-142
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    • 1998
  • Helicobacter pylori is a Gram-negative bacterium which causes chronic gastritis and is associated with gastric ulcer, duodenal ulcer and gastric carcinoma. In the process of screening of antibacterial activities against H. pylori from soil microorganisms, fungus No. 60686 was isolated. After fermentation of No.60686, the antibacterial compound was isolated, purified and followed by extraction of mycelium with organic solvents, acetone and ethyl acetate, through silica gel chromatography, LH-20 gel chromatography and HPLC. As a result of the structural analyses of the compound by IR, $^1$H- and $^{13}$C-NMR, FAB/Mass spectrophotometer, the compound having the antimicrobial activity was identified as chaetoglobosin A ($C_{32}H_{36}N_2O_5$), a cytochalasan derivative. The antimicrobial activity of chaetoglobosin A was tested against Gram-positive and negative bacteria by paper disk method. Among the test strains of 9 Gram-positive bacteria and 18 Gram-negative bacteria containing 4 H. pylori strains, the growth of 4 H. pylori strains and 3 S. aureus strains (SG 511, 285 and 503) was only inhibited by chaetoglobosin A. Also it was shown that its growth inhibition against H. pylori strains was stronger than that against S. aureus strains at the treatment of the same concentration. Therefore it was concluded that chaetoglobosin A has a specific growth inhibition against H. pylori of the tested bacteria.

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AB3217-A and B, herbicidal compounds related to anisomycin from Streptomyces sp. ME-13 (Streptomyces sp. ME-13 균주가 생산하는 anisomycin계 AB3217 화합물의 제초활성)

  • Kim, Won-Kon;Kim, Jong-Pyung;Park, Dong-Jin;Kim, Chang-Jin;Kwak, Sang-Soo;Yoo, Ick-Dong
    • Applied Biological Chemistry
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    • v.39 no.2
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    • pp.153-158
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    • 1996
  • During the screening of herbicidal substances from microbial secondary metabolites using photoautotrophic cells, a strain of ME-13 with strong herbicidal activity was isolated from soil. Based on the taxonomic studies, the strain was identified as Streptomyces. Two active compounds were purified from the culture broth through the column chromatographies using active charcoal, silica gel, MCI gel, and ODS HPLC. The compounds were identified as AB3217-A and B, respectively, related to anisomycin by spectroscopic methods. AB3217-A and B completely suppressed the germination of radish and barnyard grass at 25 ppm. In comparison to anisomycin, they showed the 6 times higher inhibitory activities against the growth of shoot and root of radish and barnyard grass with EC5O of around 6 ppm.

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Isolation and Identification of Low Molecular Volatile Compounds from Ethyl Acetate Layer of Korean Black Raspberry (Rubus coreanus Miq.) Wine (복분자(Rubus coreanus Miquel) 와인의 ethyl acetate 획분으로부터 저분자 휘발성 화합물들의 분리 및 구조해석)

  • Cho, Jeong-Yong;Kim, Seong-Ja;Lee, Hyoung-Jae;Kim, Jin-Young;Lym, Ik-Jae;Kang, Seong-Koo;Park, Keun-Hyung;Moon, Jae-Hak
    • Korean Journal of Food Science and Technology
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    • v.43 no.5
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    • pp.558-563
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    • 2011
  • In the course of our investigation for chemical constituents in the ethyl acetate layer of Korean black raspberry wine, five compounds were isolated and purified by silica gel column chromatography and high-performance liquid chromatography. The isolated compounds were identified as ethyl succinate (1), vanillic acid (2), ethyl 3,4-dihydroxybenzoate (3), furan-2-ol (4), and 4-(4-hydroxyphenyl)butan-2(S)-ol (5) based on the spectroscopic data of electrospray ionization tandem mass spectrometry and nuclear magnetic resonance. The presence of 2 in Korean black raspberry has previously reported. However, 1 and 3-5 in Korean black raspberry and its wine were isolated for the first time.

Identification of NMR Data for ginsenoside Rg1 (Ginsenoside Rg1의 NMR 데이터 동정)

  • Lee, Dae-Young;Cho, Jin-Gyeong;Lee, Min-Kyung;Lee, Jae-Woong;Park, Hee-Jeong;Lee, Youn-Hyung;Yang, Deok-Chun;Baek, Nam-In
    • Journal of Ginseng Research
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    • v.32 no.4
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    • pp.291-299
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    • 2008
  • The fresh ginseng roots were extracted in aqueous methanol (MeOH), and the obtained extracts were partitioned using ethyl acetate (EtOA), n-butanol (n-BuOH), and water, successively. The repeated silica gel column chromatography for n-BuOH fraction afforded a purified ginsenoside $Rg_1$. The physico-chemical, spectroscopic and chromatographic data of ginsenoside $Rg_1$, such as crystallization characteristics, melting point, specific rotation, infrared spectrometry (IR) data, fast atom bombardment/mass spectrometry (FAB/MS) data, nuclear magnetic resonance (NMR) data, retention factor (Rf) in thin layer chromatography (TLC) experiment, and retention time (r.t.) in HPLC analysis, were measured and compared with those reported in literatures. Especially, the previous literatures reported different data for ginsenoside $Rg_1$ in the $^{1}H-$ and $^{13}C$-NMR experiments. This paper gives the exactly assigned NMR data through 2D-NMR experiments, such as $^{1}H-^{1}H$ correlation spectroscopy (COSY), hetero nuclear single quantum correlation (HSQC), and hetero nuclear multiple bond connectivity (HMBC).

Constituents of Antimutagenic Factor from Brown Rice (현미의 항돌연변이 활성물질의 구성성분)

  • Chun, Hyang-Sook;Moon, Tae-Wha;Kim, In-Ho
    • Applied Biological Chemistry
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    • v.38 no.5
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    • pp.478-483
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    • 1995
  • To investigate the constituents of antimutagenic factor from brown rice, methanol extracts were fractionated into ether, ethylacetate, buthanol and aqueous fractions. The ether fractions showed distinct antimutagenic effect and active spot were selected by silica gel chromatography. The specific activity of active spot decreased with isolation of the active components from the methanol extract. Qualitative analyses of the active spot by using various spraying reagents revealed that ninhydrin and orcinol did not develop colored reactions. But, ferric chloride, 2,7-dichlorofluorescein, antimony pentachloride, phosphomolybdic acid, bromothymol blue and rhodamine 6G led to colored reactions. These results suggested that the consitituents of active material were neither polar nor nitrogen-containing compounds and that they may contain phenolic compounds and fatty acid derivatives. Main compounds of the active spot were analyzed to be o-hydroxy benzyl alcohol(saligenin), octanoic acid(caprylic acid), 9,12-cis-octadecadienoic acid(linoleic acid), 11-cis-octadecenoic acid(oleic acid), hexadecanoic acid(palmitic acid), 1H-indole-2-carboxylic acid and 1,2-benzenedicarboxylic acid(phthalate) in GC/Mass spectrum, and antimutagenicity of these active compounds using standard regeant was reconfirmed in S. typhimurium reversion assay.

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Identification of Antioxidative Components from Ethanol Extract of Rhus verniciflua STOKES (옻나무 에탄올 추출물로부터 항산화 활성 물질의 구조동정)

  • Kim, In-Won;Shin, Dong-Hwa;Baek, Nam-In
    • Korean Journal of Food Science and Technology
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    • v.31 no.6
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    • pp.1654-1660
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    • 1999
  • The free phenolic acid fraction of the chloroform extract from 75% ethanol extract of Rhus verniciflua STOKES (RCF) showed stronger antioxidative activity than BHT, BHA and ${\delta}-tocopherol$ at the same concentration. RCF components were isolated and identified by silica gel column chromatography, thin layer chromatography, mass spectrometer and $^1H-NMR\;and\;^{13}C-NMR$. The antioxidative activity was confirmed by electron donating activity, Rancimat method and thiobarbituric acid test in liposome system. RCF-11 could be further separated into three fractions. The antioxidative active compounds were purified and identified as gallic acid, butin and butein. The RCF-13 was purified and identified as sulfuretin.

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Isolation and Identification of Antimicrobial Compound from Jakyak(Paeonia japonica var. pilosa N$_{AKAI}$ (백작약으로부터 식품부패 미생물에 대한 항균성 물질의 분리 및 동정)

  • 황재선;전희정;한영실
    • Korean journal of food and cookery science
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    • v.16 no.5
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    • pp.445-452
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    • 2000
  • Antimicrobial activity of Jakyak(Paeonia japonica var. pilosa $N_{AKAI}$) was investigated. Methanol extract of dried Jakyak was fractionated to hexane, chloroform, ethylacetate, butanol and aqueous fraction. Ethylacetate fraction among these fractions showed the highest inhibitory effect on the microorganisms such as B. subtilis, L. monocytogenes, and V. parahaemolyticus at 500 $\mu\textrm{g}$/disc. Ethylacetate fraction was further fractionated into 11 fractions by silica gel column and thin layer chromatography(TLC). The results showed that ethylacetate fractions No. 3, 4 and 5 had the highest antimicrobial activity. They were mixed again, re-separated, and 5 fractions were obtained. Among them, the highest inhibitory effect was obtained in No. 3 fraction, which was identified as cetyl alcohol by HPLC and GC-MS.S.

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Characteristics of Nitrite Scavenger Derived from Seeds of Cassia tora L. (결명자 추출물의 아질산염 소거인자의 특성)

  • Park, Yeung-Beom;Lee, Tae-Gee;Kim, Oi-Kyung;Do, Jeong-Ryong;Yeo, Saeng-Gyu;Park, Yeung-Ho;Kim, Seon-Bong
    • Korean Journal of Food Science and Technology
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    • v.27 no.1
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    • pp.124-128
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    • 1995
  • The present study was conducted to elucidate nitrite scavenger derived from seeds of Cassia tora L. Methanolic extract of Cassia tora L was refractionated into ethyl ether, chloroform, ethyl acetate and water farction, and nitrite scavenging abilities of these fractions were investigated. Among these fractions, ethyl acetate fraction had the strongest nitrite scavenging ability(97%/2 mg). Therefore, to further investigate nitrite scavenger, ethyl acetate fraction was fractionated by silica gel column chromatography with a chloroform-methanol($10:0{\sim}0:10$) and then compound A and B were isolated. Compound A had a stronger nitrite-scavenging effect than compound B. Therefore the further separation of compound A was carried out by HPLC(32% acetonitrile. 1 ml/min) using ${\mu}Bondapak$C18 column$(3.9{\times}300 mm)$ and finally compound A-1 was obtained from compound A. Compound A-1 had by far nitrite-scavenging ability as compared with that of ascorbic acid. Compound A-1 was identified as $nor-rubrofusarin-6-{\beta}-mono-D-glucoside$ from the profiles of UV, IR and $^1H-NMR$.

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Isolation and Identification of α-Glucosidase Inhibitors from Morus Root Bark (상백피로부터 α-Glucosidase 저해제의 분리 및 동정)

  • Jang, Yeong Jung;Leem, Hyun Hee;Jeon, Yeong Hee;Lee, Dong Hee;Choi, Sang Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.7
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    • pp.1090-1099
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    • 2015
  • Among the four different parts of mulberry (Morus alba L.) tree, ethanol extract of Morus root bark showed the highest ${\alpha}$-glucosidase inhibitory activity ($IC_{50}=12.01{\mu}g/mL$). Bioassay-guided fractionation of the ethanolic extract of root bark by Diaion HP-20, silica gel, ODS-A, and Sephadex LH-20 column chromatographies led to the isolation of four compounds, including Compound (Comp.) 1 ($IC_{50}=5.22{\mu}g/mL$), Comp. 2 ($IC_{50}=1.78{\mu}g/mL$), Comp. 3 ($IC_{50}=2.94{\mu}g/mL$), and Comp. 4 ($IC_{50}=1.54{\mu}g/mL$) with strong ${\alpha}$-glucosidase inhibitory activities. Their chemical structures were elucidated as morusin (Comp. 1), kuwanon H (Comp. 2), chalcomoracin A (Comp. 3), and chalcomoracin B (Comp. 4) by UV and NMR spectral analyses. These results suggest that prenylflavonoid and mulberrofuran of Morus root bark may be useful as potential therapeutic agents for diabetes.

Purification and Characteriztion of an Antifungal Antibiotic from Bacillus megaterium KL 39, a Biocontrol Agent of Red-Papper Phytophtora Blight Disease. (고추역병균 Phytophthora capsici를 방제하는 길항균주 Bacillus megaterium KL39의 선발과 길항물질)

  • 정희경;김상달
    • Microbiology and Biotechnology Letters
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    • v.31 no.3
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    • pp.235-241
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    • 2003
  • For the biological control of Phytophthora blight of red-pepper caused by Phytophthora capsici, an antibiotic-producing plant growth promoting rhizobacteria (PGPR) Bacillus sp. KL 39 was selected from a local soil of Kyongbuk, Korea. The strain KL 39 was identified as Bacillus megaterium by various cultural, biochemical test and API and Microlog system. B. megaterium KL 39 could produce the highest antifungal antibiotic after 40 h of incubation under the optimal medium which was 0.4% fructose, 0.3% yeast extract, and 5 mM KCl at 30 C with initial pH 8.0. The antifungal antibiotic KL 39 was purified by Diaion HP-20 column, silica gel column, Sephadex LH-20 column, and HPLC. Its RF value was confirmed 0.32 by thin-layer chromatography with Ethanol:Ammonia:Water = 8:1:1. The crude antibiotic KL39 was active against a broad range of plant pathogenic fungi, Rhizoctonia solani, Pyricularia oryzae, Monilinia fructicola, Botrytis cinenea, Alteranria kikuchiana, Fusarium oxysporum and Fusarium solani. The purified antifungal antibiotic KL39 had a powerful biocontrol activity against red-pepper phytophthora blight disease with in vivo pot test as well as the strain B. megaterium KL 39.