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

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Isolation and Characteristics of Prodigiosin-like Red Pigment Produced by Serratia sp. KH-95. (Serratia sp. KH-95가 생산하는 Prodigiosin계 적색 색소의 분리 및 특성)

  • 김창호;김성호;홍석인
    • Microbiology and Biotechnology Letters
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    • v.26 no.4
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    • pp.283-289
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    • 1998
  • A bacterial strain KH-95 producing a high concentration of red pigment was isolated from the soil. The strain KH-95 was identified as a strain of Serratia sp. based on morphological and physiological characteristics. The optimal temperature and initial pH range for the production of pigment were 28$^{\circ}C$ and 7.0-8.0, respectively. The red pigment was purified through solvent extraction and silica gel column chromatography. Analyzing the structure of this pigment by instrumental analysis, it was identified as prodigiosin-like compound. In optimization of carbon and nitrogen sources, all carbon sources tested in this work inhibited the production of pigment except oils. Casein fumed out to be the most suitable nitrogen source for pigment production. Other nitrogen sources such as yeast extract, beef extract and peptone showed good cell growth but potently inhibited the production of pigment.

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Allium Jesdianum Extract Improve AcetaminophenInduced Hepatic Failure through Inhibition of Oxidative/Nitrosative Stress

  • Sohrabinezhad, Zohreh;Dastan, Dara;Asl, Sara Soleimani;Nili-Ahmadabadi, Amir
    • Journal of Pharmacopuncture
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    • v.22 no.4
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    • pp.239-247
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    • 2019
  • Objectives: Allium jesdianum (Aj) is a medicinal plant that has highlighted pharmacological features. In this study, the effects of Aj extract were examined on acetaminophen (APAP)-induced hepatic failure in rats. Methods: Methanolic fraction of hydro-alcoholic extract of Aj was obtained by silica gel column chromatography method. Animals were randomly divided into four groups each containing six rats and treated by gavage as follows: the first and second groups received normal saline, the third and fourth groups were received with 50 and 100 mg/kg of Aj extract, respectively. After two consecutive weeks, the groups 2-4 were given a single dose of APAP (2 g/kg). After 48 hours, blood and liver samples were collected for biochemical and histological examinations. Results: The findings of the study demonstrated that APAP caused a significant increase in ALT (P < 0.001), AST (P < 0.001), LDH (P < 0.001), ALP (P < 0.001) serum levels, hepatic lipid peroxidation (LPO; P < 0.001) and nitric oxide (NO; P < 0.001). In this regard, APAP led to the depletion of the total antioxidant capacity (TAC; P < 0.001), glutathione and total thiol groups (TTGs; P < 0.001), and structural change in the liver. In the Aj extract groups, a considerable improvement was found in the hepatic function alongside the histopathologic changes. Conclusion: This investigation indicated that the influential effects of Aj extract in APAP-induced hepatic failure might depend on its effect on improving oxidant/antioxidant balance in hepatic tissue.

Isolation and Identification of Bioactive Compounds from the Tuber of Brassica oleracea var. gongylodes

  • Prajapati, Ritu;Seong, Su Hui;Kim, Hyeung Rak;Jung, Hyun Ah;Choi, Jae Sue
    • Natural Product Sciences
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    • v.26 no.3
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    • pp.214-220
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    • 2020
  • Brassica oleracea var. gongylodes (red kohlrabi) is a biennial herbaceous vegetable whose edible bulbotuber-like stem and leaves are consumed globally. Sliced red kohlrabi tubers were extracted using methanol and the concentrated extract was partitioned successively with dichloromethane (CH2Cl2), ethyl acetate (EtOAc), n-butanol (n-BuOH) and water (H2O). Repeated column chromatography of EtOAc fraction through silica, sephadex LH-20 and RP-18 gel led to isolation of eleven compounds of which compound 1 was a new glycosylated indole alkaloid derivative, 1-methoxyindole 3-carboxylic acid 6-O-β-D-glucopyranoside. Others were known compounds namely, β-sitosterol glucoside (4), 5-hydroxymethyl-2-furaldehyde (5), methyl-1-thio-β-D-glucopyranosyl disulfide (6), 5-hydroxy-2-pyridinemethanol (7), (3S,4R)-2-deoxyribonolactone (8), n-butyl-β-D-fructopyranoside (9), uridine (10) and three fructose derivatives, D-tagatose (11), β-D-fructofuranose (12) and β-D-fructopyranose (13). Similarly, isolation from CH2Cl2 fraction gave two known indole alkaloids, indole 3-acetonitrile (2) and N-methoxyindole 3-acetonitrile (3). The structure elucidation and identification of these compounds were conducted with the help of 13C and 1H NMR, HMBC, HMQC, EIMS, HR-ESIMS and IR spectroscopic data, and TLC plate spots visualization. Compounds 2, 3, 4, 5, 6, 7, 8 and 9 are noted to occur in kohlrabi for the first time. Different bioactivities of these isolated compounds have been reported in literature.

A New Compound Isolation and Structure Analysis from Phellodendron Amurense Fruit Extract (황벽나무 열매 추출물로부터 신규 화합물의 분리 및 구조분석)

  • Kim, Young-Hee;Choi, Jung Eun;Hong, Jin-Young;Jo, Chang Wook;Lee, Jeung-Min;Kim, Soo Ji
    • Journal of the Korean Wood Science and Technology
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    • v.41 no.4
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    • pp.269-275
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    • 2013
  • Antifungal and insecticidal activity of Korean traditional medicinal plants was carried out to develop natural material for the development of organic cultural heritage conservation. As a result, Phellodendron amurense fruit was finally selected as a candidate of antifungal and insecticidal natural material. An novel active compound was purified from the ethylacetate fraction of Phellodendron amurense fruits using silica gel and Sephadex LH-20 column chromatography and PTLC. The compound was obtained as yellow oil form; UV ${\lambda}_{max}$(MeOH): 260 nm. The chemical structure of novel compound was determined as (4'-ethyl-2'-methylfuranyl)-6-methoxy-7-methylnona-2E,4E,6Z,8E-tetraenoic acid on the basis of various NMR experiments including $^1H$- and $^{13}C$-NMR, HMQC, HMBC and ESI-mass spectrum.

Purification and Characterization of CDMHK, a Growth Inhibitory Molecule Against Cancer Cell Lines, from Myxobacterium sp. HK1 Isolated from Korean Soil

  • LEE HAN-KI;LEE IN-HYE;YIM JEE-SUN;KIM YONG-HO;LEE SANG-HEE;LEE KISAY;KOO YOON-MO;KIM SANG-JIN;JEONG BYEONG-CHUL
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.734-739
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    • 2005
  • Myxobacterium sp. HK1, isolated from Korean soil, degrades cellulose, differentiates to fruiting body, and its 16s rDNA has $95\%$ similarity to Polyangium sp. An anticancer molecule, CDMHK, was identified from culture broth of Myxobacterium sp. HK1, and purified by Diaion HP20, Silica gel, Sephadex LH-20 chromatography, and preparative HPLC using an YMC OSD-A C18 column. The molecular structure and formula were determined to be $C_{l2}H_{l9}N_3O_2$ (M.W 237) by MS spectrometry, 300 MHz $^{1}H\;and\;^{13}C$ NMR. The CDMHK was not active against Escherichia coli, Staphylococcus aureus, and Candida albicans. However, this molecule inhibited the growth of various cancer cell lines. The $ED_{50}$ values of CDMHK were determined to be 0.147, 0.086, 0.18, 0.166, and 0.142 $\mu$g/ml against A549, SK-OV-3, SK-MEL-2, VF498, and HCTl5 cancer cell lines, respectively. In addition, the CDMHK was able to induce apoptosis of the CCRF-CEM cancer cell line, evidenced by DNA fragmentation assay and DAPI staining.

Evaluation of Environmental Mutagens-Complex Mixture in Diesel Exhaust Respirable Particulate Matter

  • Kim, Soung-Ho;Ryu, Byung-Tak;Jang, Hyoung-Seok;Kim, Yun-Hee;Lee, Do-Han;Han, Kyu-Tae;Oh, Seung-Min;Chung, Kyu-Hyuck
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.05a
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    • pp.194-194
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    • 2003
  • The International Agency for Research on Cancer (IARC, 1989) has classified whole diesel exhaust as probably carcinogenic to humans. Diesel exhaust particulate matter (DPM) adsorbs different chemical substances including PAHs and nitroarenes. DPM is emphasized because it is a major component of diesel exhaust, it is suspected of contributing to a health hazard. Diesel exhaust is a complex mixture of carbon particles and associated organics and inorganics, and it is not known what fraction or combination of fractions cause the health effects [cancer effects, noncancer effects (respiratory tract irritation/inflammation and changes in lung function)] that have been observed with exposure to diesel exhaust. In order to identify which chemical classes are responsible for the majority of the observed biological activities, we performed a particular biological/chemical analysis. Respirable particulate matter (PM2.5: <2.5mm) was collected from diesel engine exhaust using a high-volume sampler equipped with a cascade impactor. Particulate oganic matter was extracted by the dichloromethane/sonication method and the crude extract was fractionated according to EPA recommended procedure into seven fractions by acid-base partitioning and silica gel column chromatography. We examined genotoxic potentials of diesel exhaust particulate matter using novel genotoxicity tests, which are rapid, simple and sensitive methods for assessing DNA-damage at the DNA and chromosomal level (comet assay, in vitro MN test and Ames test). Higher genotoxic potency was observed in non polar fractions and several PAHs were detected by GC-MS, such as 1,2,5,6 dibenzanthracene, chrysene, 1,2-benzanthracene, phenanthrene and fluoranthene.

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Isolation of Natural Antioxidants from the Root of Zingiber officinale R. (생강(Zingiber officinale R.)근경으로부터 항산화 활성물질의 분리)

  • Bang, Myun-Ho;Song, Jung-Choon;Kim, Sun-Lim;Hur, Han-Sun;Baek, Nam-In
    • Applied Biological Chemistry
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    • v.44 no.3
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    • pp.202-205
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    • 2001
  • The root of Zingiber officinale R. was extracted with 80% aqueous MeOH, and the concentrated extract was fractionated with EtOAc, n-BuOH and water phases. The EtOAc fraction exhibited most potent antioxidant activity among them, and two active compounds were isolated by activity-guided separation using repetitive silica gel column chromatography. On the bases of spectral data and the chemical characteristics, the structure of the compounds were determined as 6-shogaol and 12-shogaol, respectively. The $EC_{50}$ values of them were evaluated as 26.4 ${\mu}M$ and 22.5 ${\mu}M$, respectively, where these values were higher than the commercial antioxidants, such as, BHA, BHT and ${\alpha}-tocopherol$.

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Presence of cis-11, 12-Methylene Octadecanoic Acid in the Oils of Ternstroemia gymnanthera (후피향 종실의 cis-11, 12-Methylene Octadecanoic acid에 관한 연구)

  • Kim, Seong-Jin;Joh, Yong-Goe;Lim, Hee-Ryeong;Choi, Eun-Jin;Kim, Tae-Sook
    • Korean Journal of Food Science and Technology
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    • v.23 no.1
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    • pp.68-75
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    • 1991
  • The seed oil of Ternstroemia gymnanthera. a species of the Ternstroemiaceae, is mainly composed of triglyceride(92.4%), followed by polar lipids(5.9%), sterol(1.2%) and pigments(0.5%). This oil contains 4.8% of cis-11, 12-methylene octadecanoic acid(lactobacillic acid) in the fatty acid composition of the total oil. This identification is based on information from non-urea inclusion formation, silver nitrate impregnated silica gel column and gas liquid chromatography, $^1H-&\;^{13}C-nuclear$ magnetic resonance and mass spectroscopy. Smaller amounts(0.1%) of presumptive 9, 10-methylene hexadecanoic acid(dihydro malvalic acid) is also detected. The major fatty acids in this oil are C18 : 1(36.1%), C18: 2(30.9%), C16: 0(15.1%), C16: 1(7.6%) and C18: 0(3.4%).

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Anti-proliferative and Antioxidant Activities of 1-methoxy-3-methyl-8-hydroxy-anthraquinone, a Hydroxyanthraquinoid Extrolite Produced by Amycolatopsis thermoflava strain SFMA-103

  • Kumar, C. Ganesh;Mongolla, Poornima;Chandrasekhar, Cheemalamarri;Poornachandra, Yedla;Siva, Bandi;Babu, K. Suresh;Ramakrishna, Kallaganti Venkata Siva
    • Microbiology and Biotechnology Letters
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    • v.45 no.3
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    • pp.200-208
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    • 2017
  • Actinobacteria are prolific producers of a large number of natural products with diverse biological activities. In the present study, an actinobacterium isolated from sunflower rhizosphere soil sample collected from Medak, Andhra Pradesh, South India was identified as Amycolatopsis thermoflava strain SFMA-103. A pigmented secondary metabolite in culture broth was extracted by using methanol and it was further purified by silica gel column chromatography with methanol-chloroform solvent system. Structural elucidation studies based on UV-visible, 1D and 2D-NMR, FT-IR, and mass spectroscopic analyses confirmed the structure as 1-methoxy-3-methyl-8-hydroxy-anthraquinone. It showed significant in vitro anticancer activity against lung cancer and lymphoblastic leukemia cells with $IC_{50}$ values of 10.3 and $16.98{\mu}M$, respectively. In addition, 1-methoxy-3-methyl-8-hydroxy-anthraquinone showed good free radical scavenging activity by DPPH method with an $EC_{50}$ of $18.2{\mu}g/ml$. It also showed other promising superoxide radical scavenging, nitric oxide radical scavenging and inhibition of lipid peroxidation activities. This is a first report of anti-proliferative and antioxidant activities of 1-methoxy-3-methyl-8-hydroxy-anthraquinone isolated from A. thermoflava strain SFMA-103 which may find potential application in biotechnological and pharmaceutical fields.

Anticoagulation and Anticancer Constituents from Eugenia caryophyllata Thunb

  • Han, Kyung-Min;Kim, Dong-Hyun;Ahn, Eun-Mi;Lee, Youn-Hyung;Chung, In-Sik;Kim, Dae-Keun;Kwon, Byoung-Mog;Kim, Sung-Hoon;Baek, Nam-In
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.2
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    • pp.82-88
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    • 2007
  • From the EtOAc fraction of Eugenia caryophyllata, four compounds were isolated through activity-guided silica gel column chromatography, From the result of spectroscopic data including NMR, MS and IR, the chemical structures of the compounds were determined as 1-allyl-4-hydroxy-3-methoxybezene acetate (eugenol acetate, 1), 1-allyl-4-hydroxy-3-methoxybezene (eugenol, 2), $3{\beta}-hydroxyolean-12-en-28-oic$ acid (oleanolic acid, 3) and $2{\alpha}$, $3{\beta}-dihydroxyolean-12-en-28-oic$ acid (maslinic acid, 4). Compounds 3 and 4 were isolated for the first time from this plant. Also, compounds 1, 2 and 3 exhibited relatively high platelet aggregation inhibitory activity with the $IC_{50}$ values of 0.24, 0.09 and 0.07 mM, respectively. Compound 2 significantly prolonged activated partial thromboplastin time (aPTT) with the value of $124{\pm}11.2$ seconds as compared to the control with the value of $37.5{\pm}2.2$ seconds at the concentration of 50 ${\mu}g/ml$. Compounds 1 and 3 revealed inhibitory activity on farnesyl protein transferase (FPTase) with the $IC_{50}$ values of 0.49 and 0.24 mM and compounds 1 and 2 highly inhibited the growth of rat-H-ras cells with the $Gl_{50}$ values of 6.63 and 5.70 ${\mu}M$, respectively.