• Title/Summary/Keyword: Flash Column Chromatography

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Isolation and Structural Analysis of Acetyl Soyasaponin $A_1$ from Hypocotyl of Soybean (콩 Hypocotyl에서 Acetyl Soyasaponin $A_1$의 분리 및 구조 분석)

  • Kim, Sun-Lim;Bang, Myun-Ho;Kim, Jung-Tae;Chi, Hee-Youn;Chung Ill-Min;Kim, Hyun-Bok;Berhow Mark A.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.spc1
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    • pp.166-173
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    • 2006
  • Soyasaponins are phytochemicals of major interest fur their health benefits. Chemical investigation of a soybean phytochemical concentrate resulted in the isolation and identification of triterpenoid saponins. The MeOH extraction of defatted hypocotyl separated from soybeans was peformed by the automated solvent extractor (ASE). Fractionation was performed on a flash column ($150mm{\times}40mm$ i.d.) packed with a preparative $C_{18}$ reverse phase bulk packing material $(125\AA,\;55-105{\mu}m)$ and monitored at 210 nm, and collected 14 fractions. Consequent Fsat preparative column liquid chromatography (Fast PCLC) was performed for the purification of Fraction-I (Fr-I) collected from the fraction 8 and 9 of flash chromatography. Fsat PCLC was performed on a Luna $C_{18}\;10{\mu}m,\;100{\AA}$, semipreparative reverse phase column ($250cm{\times}50mm$ i.d.) for the purification of isolated unknown compound (Fr-I-2). Chemical structure of acetyl soyasaponin $A_1\;(MW:1436.6,\;C_{67}H_{104}O_{33})$ was identified and determined by a combination of extensive NMR ($^1H-NMR$, 400 MHz; $^{13}C-NMR$, 100 MHz; DEPT), IR, UV, and ESI-MS analysis.

Rapid Separation of Cellular Cyclosophoraoses Produced by Rhizobium Species

  • Seo, Dong-Hyuk;Lee, Sang-Hoo;Park, Hey-Lin;Kwon, Tae-Jong;Jung, Seun-Ho
    • Journal of Microbiology and Biotechnology
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    • v.12 no.3
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    • pp.522-525
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    • 2002
  • A very rapid and efficient separation technique for cellular rhizobial cyclosophoraoses was developed based on fractional precipitation and partition chromatography. Cyclosophoraoses are known to function in the osmotic regulation and root nodule formation of legumes during the nitrogen fixation process. Cyclosophoraoses are produced as unbranched cyclic (1longrightarrow12)-${\beta}$-D-glucans in Agrobacterium or Rhizobium species. Recent research has shown that cyclosophoraoses can form inclusion complexation with various unstable or insoluble guest chemicals, thereby implying great potential for industrial application. Typical separation of pure cellular cyclosophoraoses has been so far carried out by several time-consuming steps, including size exclusion, anion exchange, and desalting liquid chromatographies, with a relatively poor recovery. However, the proposed method demonstrated that the successive application of fractional ethanol precipitation and one step of silica gel-based flash column chromatography was enough to simultaneously purify neutral or anionic forms of cyclosophoraoses. This novel technique is very rapid and provides a high recovery.

Isolation and Characterization of Allelopathic Substances from Sorghum Stem (수수 줄기에 함유(含有)된 타감물질(他感物質)의 분리(分離) 및 특성(特性) 구명(究明))

  • Kim, S.Y.;De Datta, S.K.;Robles, R.P.;Kim, K.U.;Lee, S.C.;Shin, D.H.
    • Korean Journal of Weed Science
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    • v.14 no.2
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    • pp.156-162
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    • 1994
  • To better understand the exact nature of the major toxic compound responsible for phytotoxicity of sorghum stem, the most toxic compound from the stem extract was isolated by rapid chromatography and subsequently purified by thin-layer chromatography(TLC) and high pressure liquid chromatography(HPLC). Of the eight fractions isolated by rapid chromatography, the fraction with solvent combinations of butanol (8) : acetic acid (1) : water (1) had the highest toxicity. Further separation of the fraction by TLC in a solvent mixture of butanol (24) : acetic acid (16.4) : water (7) : propanol (1) showed that the spot with an $R_f$ 0.71 had one major peak with retention time of 20.40 minutes. Upon subjecting gas chromatography and the HPLC fraction to the mass spectrometry, the toxic compound is probably one of the four compounds ; 1-methyl-1-(2-propynyl)-hydrazine, 1-aziridineethanol, 5-chloro-2-pentanone, and 2-(methylseleno)-ethanamine.

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Identification and Biological Activity of Two New Phytotoxins Isolated from Botrytis cinerea (Botrytis cinerea로부터 분리한 두 개의 새로운 phytotoxin의 구조 결정 및 생물활성)

  • Kim, Geum-Jung;Yoon, Mi-Young;Kim, Heung-Tae;Choi, Gyung-Ja;Jang, Kyoung-Soo;Choi, Yong-Ho;Park, Myung-Soo;Cha, Byeong-Jin;Kim, Jin-Cheol
    • Research in Plant Disease
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    • v.15 no.2
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    • pp.112-119
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    • 2009
  • We discovered two novel phytotoxins produced by the pathogenic fungus, Botrytis cinerea. Among the twenty-five B. cinerea isolates, which were obtained from various host plants in 1994 and 1996, twenty-two showed strong or moderate pathogenicity on five plants such as cucumber, tomato, red pepper, tobacco and Chinese cabbage. The culture filtrate of the B. cinerea 2-16 strain showed the most potent phytotoxic activity in a tobacco leaf-wounding assay. Two novel phytotoxins were isolated from the liquid cultures of B. cinerea 2-16 by ethyl acetate extraction, flash silica gel column chromatography, silica gel column chromatography, Sephadex LH-20 column chromatography, preparative TLC and subsequently preparative HPLC. Their chemical structures were determined to be 3-O-acetyl botcinol and 3-O-acetyl botcinolide, respectively, by mass and NMR spectral analyses. These two phytotoxins caused leaf necrosis in a leaf-wounding bioassay, and significant electrolyte leakage from leaf tissues of tobacco. In the two bioassays tested, 3-O-acetyl botcinol exhibited stronger phytotoxic activity than 3-O-acetyl botcinolide. This is the first report on the production of both 3-O-acetyl botcinol and 3-O-acetyl botcinolide from B. cinerea.

Isolation of Herbicidal Compounds from Pulsatilla koreana Roots (백두옹(Pulsatilia koreana Nakai) 뿌리로부터 제초활성물질의 분리)

  • 정형진
    • Korean Journal of Plant Resources
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    • v.9 no.1
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    • pp.47-54
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    • 1996
  • To search herbicidal compounds in Pulsatilla koreana Nakai, methanol extract of P. koreana roots was purified by sequences of XAD-7 column chromatography, silica gel adsorption column chromatography, silica gel flash column chromatography, preparative layer chromatography and preparative high performance liquid chromatography(Prep, HPLC).The final Prep. HPLC gave two herbicidally-active fractions. These fractions treatment at 100ppm inhibited the root length of Echinochloa crus-galli seedlings by 48% and 60% as compared with the control, respectively. Components in the two active fractions were analyzed by GC-MS Spectrometry. These compounds, which were isolated from P. koreana roots, were identified as several fatty esters, hydrocarbons, squalene, evidonol, and a diazepin analogue.

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Isolation and In vitro and In vivo Antifungal Activity of Phenylacetic acid Produced by Micromonospora aurantiaca Strain JK-1

  • Kim, Hyo-Jin;Hwang, In-Sun;Kim, Beom-Seok;Hwang, Byung-Kook
    • The Plant Pathology Journal
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    • v.22 no.1
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    • pp.75-89
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    • 2006
  • The actinomycete strain JK-1 that showed strong inhibitory activity against some plant pathogenic fungi and oomycetes was isolated from Jung-bal Mountain in Ko-yang, Korea. The strain JK-1 produced spores singly borne on sporophores and the spores were spherical and 0.9-1.2 11m in diameter. The cell wall of the strain JK-1 contained meso-diaminopimelic acid. The actinomycete strain JK-1 was identified as the genus Micromonospora based on the morphological, physiological, biochemical and chemotaxonomic characteristics. From the 168 rDNA analysis, the strain JK-1 was assigned to M aurantiaca. The antibiotic MA-1 was purified from the culture broth of M aurantiaca JK-1 using various purification procedures, such as Diaion HP20 chromatography, C18 flash column chromatography, silica gel flash column chromatography and Sephadex LH-20 column chromatography. $^{1}H-$, $^{13}C-NMR$ and EI mass spectral analysis of the antibiotic MA-1 revealed that the antibiotic MA-1 is identical to phenylacetic acid. Phenylacetic acid showed in vitro inhibitory effects against fungal and oomycete pathogens Alternaria mali, Botrytis cinerea, Magnaporthe grisea, Phytophthora capsici and yeast Saccharomyces cerevisiae at < 100 $\mug$ $ml^{-1}$. In addition, phenylacetic, acid completely inhibited the growth of Sclerotinia sclerotiorum, Bacillus subtilis, Candida albicans, Xanthomonas campestris pv. vesicatoria at < $\mug$ $ml^{-1}$. Phenylacetic acid strongly inhibited conidial germination and hyphal growth of M grisea and C. orbiculare. Phenylacetic acid showed significantly high levels of inhibitory' effect against rice blast and cucumber anthracnose diseases at 250 $\mug$ $ml^{-1}$. The control efficacies of phenylacetic acid against the two diseases were similar to those of commercial compounds tricyclazole, iprobenfos and chlorothalonil .n the greenhouse.

Isolation of Herbicidal Compound from Bulbs of Lycoris chinensis var. sinuolata K.H.Tae & S.T.Ko (진노랑상사화 인경으로부터 살초활성 물질의 분리)

  • Jang, Ho-Jin;Kim, Kun-Woo
    • Korean Journal of Weed Science
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    • v.30 no.4
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    • pp.437-444
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    • 2010
  • This study was conducted to determine the herbicidal activity of allelochemicals and identify herbicidal compounds in bulbs of Lycoris chinensis var. sinuolata. Methanol extract was purified by a series of silica gel flash column chromatography and HPLC. The final HPLC gave two active fractions and an herbicidal compound was obtained. By GC/MS analysis, the herbicidal compound was identified as montanine ($O^2$-methyl pancracine), an isoquinoline alkaloid. Montanine showed 100% of growth inhibition on the shoot and root of barnyardgrass (Echinochloa crus-galli) seedlings at $50\;{\mu}g\;mL^{-1}$ as compared with the control.

Antioxidant activity of ethanol extract and methanol fractions via column chromatography from Psidium guajava Leaf (구아바 잎 추출물 및 컬럼크로마토그래피를 이용한 메탄올 분획물의 항산화 활성)

  • Byeoung-Kyu Choi
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.266-271
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    • 2023
  • The antioxidant capacity of the Psidium guajava leaf extracted with EtOH and their MeOH fractions using column chromatography were evaluated by 1,1-diphenyl-2-picrylhydrazyl and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid radical scavenging assays, total phenolic and flavonoid content, and Superoxide dismutase (SOD) assay. To determine its utility as a functional material, the crude extract was fractionated by flash column chromatography on ODS using a stepwise elution with combinations of MeOH/H2O and then all the fractions were also investigated. In the results of antioxidant activities, the 40% and 60% MeOH fractions show the meaningful values, and then the two fractions were selected to examine the isolation and identification of the major constituents via HPLC and nuclear magnetic resonance. Further purification led to isolation of two quercetin derivatives; quercitrin (1) and isoquercetin (2). Through SOD assay, some methanol fractions via column chromatography and isolated compounds showed improved antioxidant activities compared to the extract.

Purification of Antifungal Antibiotic NH-B1 from Actinomycete NH 50 Antagonistic to Plant Pathogenic Fungi (식물병원진균에 길항효과가 있는 방선균 균주 NH50에서 항진균성 항생물질 NH-B1의 순수 분리)

  • 김현겸;김범석;문석식;황병국
    • Korean Journal Plant Pathology
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    • v.14 no.3
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    • pp.191-202
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    • 1998
  • About 300 actinomycetes were isolated from two forest and one sea-shore soil and tested for inhibitory effects on mycelial growth of six plant pathogenic fungi Magnaporthe grisea, Alternaria mali, Colletotrichum gloeosporioides, Phytophthora capsici, Fusarium oxysporum f. sp. cucumerinum, and Rhizoctonia solani. Among 300 actinomycetes tested, only 16 actinomycetes showed the antifungal activity against the test fungi. Isolate NH 50 was selected for production and purification of antifungal antibiotic substances. Actinomycete isolate NH 50 displayed the broad antifungal spectra against 11 plant pathogenic fungi. To identify actinomycete isolate NH 50, cultural characteristics on various agar media, diaminopimelic acid type, and morphological characteristics by scanning electron microscopy were examined. As a result, actinomycete isolate NH 50 was classified as a rare actinomycete that had LL-DAP type and did not produce spores. After incubation of isolate NH 50 in yeast extract-malt extract-dextrose broth, antifungal compound NH-B1 that inhibited mycelial growth of some plant pathogenic fungi was purified from the methanol eluates of XAD-16 resins by a series of purification procedures, i.e., silica gel flash chromatography, C18 flash chromatography, Sephadex LH-20 column chromatography, silica gel medium pressure liquid chromatography (MPLC), C18 MPLC, and high pressure liquid chromatography (HPLC). UV spectrum and 1HNMR spectrum of antifungal compound NH-B1 dissolved in methanol were examined. The antibiotic NH-B1 showed the major peaks at 230 and 271.2nm. Based on the data of 1H-NMR spectrum, NH-B1 was confirmed to be an extremely complex polymer of sugars called polysaccharides. The antibiotic NH-B1 showed strong antifungal activity against Alternaria solani and Cercospora kikuchi, but weak activity against M. grisea.

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Anti-cancer and Anti-inflammatory Effects of Mung Bean and Soybean Extracts (녹두 및 대두추출물의 항암 및 항염증 활성)

  • Imm, Jee-Young;Kim, Seok-Joong
    • Korean Journal of Food Science and Technology
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    • v.42 no.6
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    • pp.755-761
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    • 2010
  • The quinone reductase (QR) inducing activities of mung bean and soybean solvent extracts were compared using murine hepatoma cells (Hepa 1c1c7). The mung bean extracts (ethylacetate and ethanol) showed higher chemoprevention index values (7.88-8.22) than those of soybean extracts (2.9-5.2) from four different cultivars. The mung bean extracts also had significantly higher inhibitory effects (47-62% at 100 ${\mu}g$/mL) than the soybean extracts (15-42% at 100 ${\mu}g$/mL) against the production of nitric oxide and prostaglandin E2 in lipopolysccharide stimulated macrophage RAW264.7 cells without cytotoxicity. Among seven recovered fractions of mung bean ethanol extract obtained by C 18 silica flash column chromatography, the most non-polar fraction exhibited the highest chemoprevention index of 10.4.