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

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Antioxidation Activities of Organic Solvent Fractions Obtained from Seaweed, Hizikia fusiformis (톳(Hizikia fusiformis)에서부터 분리된 유기용매 분획물의 항산화활성)

  • Kim, Min-Jeong;Lee, Hye-Hyeon;Seo, Min-Jeong;Kang, Byoung-Won;Park, Jeong Uck;Jeong, Yong Kee
    • Journal of Life Science
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    • v.23 no.3
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    • pp.361-367
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    • 2013
  • Hizikia fusiformis has been widely used in Oriental herbal medicine and health food. To identify antioxidation properties that contain natural bioactive substances, we investigated the distribution of active compounds existing in batches of organic solvent fractionation. A dried form of H. fusiformis was subjected to sequential fractionation using n-hexane, ethyl acetate, n-BuOH, and aqueous n-BuOH. The results showed that among the four isolated fractions, the n-BuOH fraction showed the highest antioxidation activities. The n-BuOH fraction was applied to reserve-phase silica gel column chromatography, which produced three fractions: BA, BB, and BC. Among these fractions, BB showed the highest antioxidation activities, which increased in a concentration-dependent manner. At a concentration of 1.0 mg/ml n-BuOH fraction, the activities of DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging and reducing power were approximately $45{\pm}0.14%$ and $1.34{\pm}0.23$, respectively. In addition, the activities of ${\beta}$-carotene-linoleic acid, hydrogen peroxide scavenging, and ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)) radical scavenging were $76{\pm}0.12%$, $82{\pm}0.06%$, and $65{\pm}0.17%$, respectively. These findings suggest that the BB fraction contains potent antioxidation properties and that it could be used in the production of natural and functional foods.

Polyacetylene Compounds from Panax ginseng C.A. Meyer (인삼의 Polyacetylene 화합물)

  • Shim Sang Chul;Chang Suk-Ku
    • Proceedings of the Ginseng society Conference
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    • 1988.08a
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    • pp.122-128
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    • 1988
  • Several major polyacetylene compounds were isolated from the petroleum-ether fraction of fresh Korean ginseng roots through solvent fractionation. partition and silica gel column chromatography. Further separation of acetylenic compounds was accomplished by bonded normal phase HPLC utilizing a moderately nonpolar microparticulate column. The preparative separation for the various spectral measurements was carried out by low pressure preparative liquid chromatography. The chemical structure of these polyacetylenes separated was determined by UV. IR/FTIR. $^{1}H$ NMR. mass spectral and elemental analysis. These are identified to be heptadeca-1-en-4.6-diyn-3.9.l0.-triol [1] heptadeca-1.9-dien-4.6-diyn-3-ol. heptadeca-1.8-dien-4.6-diyn-3.10-diol and the 4th was denatured polyacetylene. heptadeca-1.4-dien-6.8-diyn-3.10-diol. Two different p-substituted benzoates of panaxynol were synthesized for the determination of exciton chirality. The circular dichroism spectra in the UV region show that panaxynol p-bromobenzoate and p-dimethyl-aminobenzoate constitute negative exciton chirality [2]. Isolated major polyacetylene compounds were irradiated in aerated solution with 300 nm UV light to obtain the oxidized product at the allylic alcohol center to corresponding carbonyl compounds such as heptadeca-1-en-4.6-diyn-9.10-diol-3-one and heptadeca-1.9-dien-4.6-diyn-3-one. These photooxidation compounds have en-on-diyne chromophore and undergo nucleophilic addition reaction with methanol to yield ${\beta}-methoxy$ carbonyl compounds such as heptadeca-9-en-4.6-diyn-1-methoxy-3-one and heptadeca-4.6-diyn-1-methoxy-9.10-diol-3-one.

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A Tuber Lectin from Arisaema jacquemontii Blume with Anti-insect and Anti-proliferative Properties

  • Kaur, Manpreet;Singh, Kuljinder;Rup, Pushpinder Jai;Kamboj, Sukhdev Singh;Saxena, Ajit Kumar;Sharma, Madhunika;Bhagat, Madhulika;Sood, Sarvesh Kumar;Singh, Jatinder
    • BMB Reports
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    • v.39 no.4
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    • pp.432-440
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    • 2006
  • A tuber lectin from Arisaema jacquemontii Blume belonging to family Araceae was purified by employing a single step affinity chromatography using column of asialofetuin-linked amino activated silica beads and the bound lectin was eluted with 100 mM glycine-HCl buffer pH 2.5. The purified A. jacquemontii lectin (AJL) showed a single protein band with an apparent molecular mass of 13.4 kDa when submitted to SDS-polyacrylamide gel electrophoresis under reducing as well as non-reducing conditions. The native molecular mass of AJL determined by gel filtration on a Biogel P-200 column was 52 kDa and its carbohydrate content was estimated to be 3.40%. Thus AJL is a tetrameric glycoprotein. The purified lectin agglutinated erythrocytes from rabbit but not from human. Its activity was not inhibited by any of the mono- and disaccharides tested except N-acetyl-D-lactosamine having minimal inhibitory sugar concentration (MIC) 25 mM. Among the glycoproteins tested only asialofetuin was found to be inhibitory (MIC $125\;{\mu}g/mL$). A single band was obtained in native PAGE at pH 4.5 while PAGE at pH 8.3 showed two bands. Isoelectric focusing of AJL gave multiple bands in the pI range of 4.6-5.5. When incorporated in artificial diet AJL significantly affected the development of Bactrocera cucurbitae (Coquillett) larvae indicating the possibility of using this lectin in a biotechnological strategy for insect management of cucurbits. Larvae fed on artificial diet containing sub-lethal dose of AJL showed a significant decrease in acid phosphatase and alkaline phosphatase activity while esterase activity markedly increased as compared to larvae fed on diet without lectin. Out of various human cancer cell lines employed in sulphorhodamine B (SRB) assay, this lectin was found to have appreciable inhibitory effect on the in vitro proliferation of HCT-15, HOP-62, SW-620, HT-29, IMR-32, SKOV-3, Colo-205, PC-3, HEP-2 and A-549 cancer cell lines by 82, 77, 73, 70, 41, 41, 37, 29, 21 and 21% respectively.

Flavonol Glycosides from the Aerial Parts of Aceriphyllum rossii and Their Antioxidant Activities

  • Han Jae-Taek;Bang Myun-Ho;Chun Ock-Kyoung;Kim Dae-Ok;Lee Chang-Yong;Baek Nam-In
    • Archives of Pharmacal Research
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    • v.27 no.4
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    • pp.390-395
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    • 2004
  • The methanol extract obtained from the aerial parts of Aceriphyllum rossii (Saxifragaceae) was fractionated into ethyl acetate (EtOAc), n-BuOH and $H_2O$ layers through solvent fractionation. Repeated silica gel column chromatography of EtOAc and n-BuOH layers afforded six flavonol glycosides. They were identified as kaempferol 3-O-$\beta$-D-glucopyranoside (astragalin, 1), quercetin 3-O-$\beta$-D-glucopyranoside (isoquercitrin, 2), kaempferol 3-O-$\alpha$-L-rhamnopyranosyl $(1{\to}6)-\beta$-D-glucopyranoside (3), quercetin 3-O$\alpha$-L-rharnnopyranosyl $(1{\to}6)-\beta$-D-qlucopyrano-side (rutin, 4), kaempferol 3-O-[$\alpha$-L-rharnnopyranosyl $(1{\to}4)-\alpha$-L-rhamnopyranosyl $(1{\to}6)-\beta$-D-glucopyranoside] (5) and quercetin 3-O-[$\alpha$-L-rhamnopyranosyl $(1{\to}4)\alpha$-L-rhamnopyranosyl $(1{\to}6)\beta$-D-glucopyranoside] (6) on the basis of several spectral data. The antioxidant activity of the six compounds was investigated using two free radicals such as the ABTS free radical and superoxide anion radical. Compound 1 exhibited the highest antioxidant activity in the ABTS $\{2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)\}$ radical scavenging method. 100 mg/L of compound 1 was equivalent to $72.1\pm1.4\;mg/L$ of vitamin C, and those of compounds 3 and 5 were equivalent to $62.7\pm0.5\;mg/L$ and $54.3\pm1.3\;mg/L$ of vitamin C, respectively. And in the superoxide anion radical scavenging method, compound 5 exhibited the highest activity with an $IC_{50}$ value of $17.6{\pm}0.3{\mu}M$. In addition, some physical and spectral data of the flavonoids were confirmed.

Aburatubolactam C, a Novel Apoptosis-inducing Substance Produced by Marine Streptomyces sp. SCRC A-20

  • Bae, Myung-Ae;Yamada, Kaoru;Uemura, Daisuke;Seu, Jung-Hwan;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.8 no.5
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    • pp.455-460
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    • 1998
  • In the course of screening for new antitumor substances, a novel cytotoxic agent inducing apoptotic cell death was isolated from the culture broth of marine bacterial strain SCRC A-20. Strain SCRC A-20 was separated from a mollusk and was chemotaxonomically identified as a Streptomyces sp. The cytotoxic substance was purified by organic solvent extraction followed by silica gel column chromatography and preparative TLC. HRFAB-MS determined its molecular formula to be $C_{30}H_{40}N_2O_5$ (MW 508). The 1D and 2D NMR spectral data demonstrated that the substance has a novel lactam structure of a 20-membered macrocycle coupled with a unique acyl tetramine and bicyclo[3.3.0] octane, which includes three methyl groups, six olefinic protons, five carbonyl groups, a conjugated diene and a dienone. The substance, named aburatubolactam C, appeared to be cytotoxic for various continuously proliferating tumor cells of human and murine origins. The $IC_{50}$ values determined by MIT assay were in the range of 0.3 to $5.8\mug/ml$. When Jurkat T cells were treated with $3\mug/ml$. of aburatubolactam C, the apoptotic DNA fragmentation was detectable within 3 h, indicating that the cytotoxic effect of aburatubolactam C on tumor cells is attributable to the induced apoptosis.

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Control Efficacy of Streptomyces sp. A501 against Ginseng Damping-off and Its Antifungal Substance

  • Minh, Nguyen Van;Woo, E-Eum;Lee, Gang-Seon;Ki, Dae-Won;Lee, In-Kyoung;Lee, Sang-Yeob;Park, Kyeonghun;Song, Jaekyeong;Choi, Jae Eul;Yun, Bong-Sik
    • Mycobiology
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    • v.45 no.1
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    • pp.44-47
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    • 2017
  • Ginseng damping-off, caused by the fungal pathogens Rhizoctonia solani and Pythium sp., is a critical disease in ginseng seedling. In a continuing effort to find microorganisms with the potential of acting as a biocontrol agent against Rhizoctonia damping-off, we found that a Streptomyces sp. A501 showed significant antifungal activity against Rhizoctonia solani. In field experiment to test the efficacy of Streptomyces sp. A501 in controlling ginseng damping-off, the incidence of damping-off disease was meaningfully reduced when ginseng seeds were soaked in the culture broth of Streptomyces sp. A501 before sowing. To perform characterization of the antifungal compound, we isolated it from the culture broth of strain A501 through Diaion HP-20 and silica gel column chromatographies and preparative high-performance liquid chromatography. The structure of the antifungal compound was assigned as fungichromin by spectroscopic methods, mainly nuclear magnetic resonance and electrospray ionization-mass analysis.

Isolation, Structure Determination and Biological Activity of 25-Epi, $3{\alpha}-Carboxyacetylquercinic$ Acid in Daedalea dickinsii (띠미로버섯 중 25-Epi, $3{\alpha}-Carboxyacetylquercinic$ Acid의 분리정제, 구조결정 및 생리활성)

  • Bae, Kang-Gyu;Min, Tae-Jin
    • Journal of the Korean Chemical Society
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    • v.44 no.1
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    • pp.37-44
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    • 2000
  • In order to develope bioactive substances, dried fmit body of Daedalea dickinsii collected from Taeback mountain was extracted with ethanol, The compounds 1 and 2 were purified by solvent extraction, silica gel column chromatography and recrystallization from the ethanol extracts. Melting point, molecular weight and molar extinction coefficient of the compound 1 were estimated to be $167{\sim}168^{\circ}C$, 572 and 5,040 at 208 nm. The structure of this compound 1 has been elucidated to be 24S, 25S, $3{\alpha}-carboxyacetylquercinic$ acid using spectroscopic properties. This compound showed antibiotic activities against fungi, yeasts and bacteria and it also showed anticancer activity $IC_{50}=64.5\;{\mu}M$) against Korean stomach cancer cell. Melting point, molecular weight and molar extinction coefficient of the compound 2 were estimated to be $233{\sim}235^<\circ}$, 572 and 5,080 at 208 nm. The structure of this compound was elucidated to be 24S, 25R, $3{\alpha}-carboxyacetylquercinic$ acid. The compound 2 showed different antibiotic activities with the compound 1.

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Isolation and characterization of an antifungal substance from Burkholderia cepacia, an endophytic bacteria obtained from roots of cucumber.

  • Park, J.H.;Park, G.J.;Lee, S.W;Jang, K.S.;Park, Y.H.;Chung, Y.R.;Cho, K.Y.;Kim, J.C.
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.95.2-96
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    • 2003
  • In order to develop a new microbial fungicide for the control of vegetable diseases using endophytic bacteria, a total of 260 bacterial strains were isolated from fresh tissues of 5 plant species. After they were cultured in broth media, their antifungal activities were screened by in vivo bioassays against Botrytis cinerea(tomato gray mold), Pythium ultimum(cucumber damping-off), Phytopkhora infestans(tomato late blight), Colletotrichum orbiculare(cucumber anthracnose), and Blumeria graminis f. sp. hordei(barley powdery mildew). As the results of screening, 38 bacterial strains showed potent antifungal activities against at least one of 5 plant pathogens. A bacterial strain EB072 displayed potent disease control activities against 3 plant diseases. Among the bacterial strains with a potent antifungal activity against cucunlber anthracnose, three bacterial strains, EB054, EB151 and EB215, also displayed a potent in vitro antifungal activity against C. acutatum, a fungal agent causing pepper anthracnose. A bacterial strain EB215 obtained from roots of cucumber was identified as Burkholderia cepacia based on its physiological and biochemical characteristics and 165 rRNA gene sequence. An antifungal substance was isolated from the liquid cultures of B. cepacia EB215 strain by ethyl acetate partitioning, repeated silica gel column chromatography, and invitro bioassay, Its structural determination is in progress by various instrumental analyses.

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Studies on Screening and Isolation of Esterase Inhibitors from Soil Microorganisms (II). Isolation of Inhibitors and Associated Lipids from Streptomyces Strain DMC-498 (한국 토양균중 Esterase 저해제 검색 및 분리에 관한 연구(제 2 보) Streptomyces Strain DMC-498 균주의 저해성분 및 관련 지질의 분리)

  • 이승정;김하원;곽진환;심미자;허영근;김성원;최응칠;김병각
    • Journal of Food Hygiene and Safety
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    • v.2 no.4
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    • pp.181-189
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    • 1987
  • To find esterase inhibitors and in the metabolites of Streptomyces strain DMC-498, two active compounds were isolated from the methanol extract of the mycelia of the strain by Silica gel column chromatography and preparatory argentation TLC. These compounds were proved to show competitive inhibition. Compound B was found to consist of linoleic and oleic acids. Fifty percent inhibition concentration ($lC_{50}$) of linoleic acid was $0.045\mu\textrm{g}/ml$, whereas oleic acid exhibited no inhibitory activity. Associated lipids: isostearic acid, isostearic acid methyl ester, oleic acid methyl ester and linoleic acid methyl ester, were isolated from the same extract, showing no inhibition of the esterase. Compound A was found to be a liquid inhibitor with an alicyclic ring and two or more oxygens, its molecular weight being more than 500.

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Volatile Flavor Components of Codonopsis lanceolata Traut. (Benth. et Hook.) (더덕뿌리중의 휘발성 향기성분)

  • Park, Joon-Yung;Kim, Young-Hoi;Kim, Kun-Soo;Kwag, Jae-Jin
    • Applied Biological Chemistry
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    • v.32 no.4
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    • pp.338-343
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    • 1989
  • The volatile oil of the root of Codonopsis lanceolata Traut. (Benth. et Hook.) was isolated by steam distillation and extraction method and fractionated by silica gel column chromatography. The total volatile oil and each fractions were analyzed by GG, GC-MS and retention indices matching. A total of 50 components were identified in the volatile oil including 16 terpene and terpene alcohols, 13 hydrocarbons, 5 alcohols, 6 aldehyde and ketones, 6 acids, 2 esters and 2 miscellaneous components. The major components were n-hexanal (7.3% of total volatile oil), trans-2-hexenal (24.9%), n-hexanol (19.8%), cis-3-hexen-1-ol (5.6%) and trans-2-hexen-1-ol (29.4%).

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