• Title/Summary/Keyword: Preparative Chromatography

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Determination and Isolation of Antioxidative Serotonin Derivatives, N-(p-Coumaroyl)serotonin and N-feruoylserotonin from Safflower Seeds (홍화종자에서 항산화성 Serotonin계 화합물, N-(p-Coumaroyl)serotonin과 N-feruoylserotonin의 분리 및 정량분석)

  • Lee, Kang-Soo;Kim, Yun-Hee;Chung, Nam-Jin
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.53 no.2
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    • pp.171-178
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    • 2008
  • In the methanol extract from safflower seeds, two kinds of antioxidant were detected by preparative HPLC [$\mu$-Bondapak $C_{18}$ column ($7.8{\times}300\;mm$)]. Two unknown compounds were defined as CA and CB which had peaks at 22.1 min and 24.5 min, respectively. Antioxidant activity was measured by their scavenging ability on the stable tree radical of 1,1-diphenly-2-picrylhydrazyl (DPPH). For bulk extraction of antioxidants, the methanol extract was fractionated with hexan, chloroform, ethyl acetate and butanol. The ethyl acetate traction showing the highest DPPH radical scavenging activity was further purified by silica gel column chromatography to CA and CB. By NMR analysis, CA and CB were identified as N-(p-Coumaroyl)serotonin and N-feruoylserotonin, respectively. The content of N-(p-Coumaroyl)serotonin and N-feruoylserotonin were analyzed by reverse phase HPLC using a $\mu$-Bondapak $C_{18}$ column ($3.9{\times}300\;mm$) with linear gradient elution from 10% acetonitrile to 50% acetonitrile for 30min on UV detector at 300 nm. The contents of N-(p-Coumaroyl)serotonin and N-feruoylserotonin were 4.11 mg/g DW and 7.29 mg/g DW, respectively, and these two DPPH radical scavengers were detected only in the hull of seeds.

PLANT BIOCHEMISTRY OF GINSENG SAPONINS(III) Radioactive Studies (2). Sodium Acetate-U-$C^{14}$ Experiment

  • The Korean Society of Ginseng The Korean Society of Ginseng
    • Proceedings of the Ginseng society Conference
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    • 1974.09a
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    • pp.101-113
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    • 1974
  • The radioactive compound sodium $acetate-U-C^{14}$ (C-14 acetate) was administered to two- and four-year-old July and September American ginseng (Panax quinquefolium L.) plants and cuttings. The C-14 acetate uptake was approximately $99\%.$ The autoradiochromatograms suggest that the saponins(panaquilins) isolated by preparative thin-layer chromatography contained impurities, especially those isolated from the leaf and stem extracts. The root and fruit methanol extracts yielded relatively pure saponins. The large amounts of panaquilin B and its proximity to panaquilin C on preparative thin-layer plates resulted in some admixing. The average concentration $(\%$ plant dry weight) of semipurified saponins were high in the leaves $(13.8\%),$ compared to fruits $(9.8\%),\;stems\;(7.9\%)\;and\;roots\;(6.3\%).$ The average percentage of C-14 acetate incorporation into panaquilins was $4.8\%.$ The average percentage of C-14 acetate incorporation into panaquilins B and C was higher $(1.40\%\;and\;1.13\%,$ respectively) than that into panaquilin C, (d), G-1 and G-2 $(0.75\%,\;0.65\%,\;0.13\%\;and\;0.53\%,$ respectively). Panaquilin synthesis may be depending upon the part collection period and age of the plant. The average percentage of C-14 acetate incorporation into panaquilin B is high in roots $(0.58\%)\;and\;stems\;(0.48\%);$ that into panaquilins C and (d) high in leaves $(0.40\%\;and\;0.45\%,$ respectively); and that into panaquilin E high in roots and leaves $(0.55\%\and\;0.50\%,$ respectively). Panaquilin G-2 was synthesized in all parts of plants. The panaquilins appear to be biosynthesized more actively in July than September (exception-panaquilin G-l). Panaquilins B, C and G-1 may be biosynthesized more actively in four-year-old plants and panaquilins (d) and E more actively in two-year-old plants. The results from expectance with cuttings suggest that the panaquilins are synthesized de novo in the above-ground parts of ginseng plants, and that panaquilin G-l may be synthesized de novo in the leaf. It is known from the tissue culture studies that panaquilins are produced by leaf, stem and root callus tissues and callus-root cultures of American and Korean ginseng plants. Panaquilins may actively be synthesized de novo in most any cell or organ of the ginseng plants. It was verified that C-14 acetate was incorporated into the panaxadiol portions of the panaquilins of two-year-old plants (sp. act., 0.56 $m{\mu}Ci/mg$) and four-year-old plants (sp. act., 0.54 $m{\mu}Ci/mg$).

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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.

Inhibitors of Tyrosinase and Melanogenesis from Galla rhois

  • Kim, Hyo-Jin;Jang, Dong-Il;Park, Sang-Won
    • Preventive Nutrition and Food Science
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    • v.2 no.4
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    • pp.285-290
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    • 1997
  • Previously, a 50% aqueous methanol extract of Galla rhois was shown to be the most potent tyrosinase inhibition activity with an {TEX}$IC_{50}${/TEX}(the concentration causing 50% inhibition of tyrosinase activity) of 0.2mg/ml of 205 crude drug extracts. To isolate tyrosinase inhibitors, the methanol extract was evaporated to a small volume in vacuo, and then partitioned stepwise with benzene and ethyl acetate(EtOAc). the EtOAc fraction was solubilized in 10% MeOH solution, and then fractionated successively by Diaion HP-20 and Sephadex LH-20 column chromatography, and preparative HPLC. Three phenolic compounds were isolated, and characterized as gallic acid(GA), methyl gallate(MG) and 1,2,3,4,6-penta-O-galloyl-$\beta$-D-glucose(PGG) by UV, IR, {TEX}${1}^H${/TEX}-&{TEX}${13}^C${/TEX}-NMR, and FAB-MS spectroscopy, PGG({TEX}$IC_{50}${/TEX}=50$\mu\textrm{g}$/ml) showed a considerable inhibitory effect against mushroom tyrosinase, while GA({TEX}$IC_{50}${/TEX}=1.6mg/ml) and MG({TEX}$IC_{50}${/TEX}=234$\mu\textrm{g}$/ml) did not show an appreciable effect. Meanwhile, MG inhibited greatly melanogenesis in a murine melanocyte cell line, Mel-Ab. MG and PGG showed typical noncompetitive inhibition patterns against mushroom tyrosinase. These results suggest that PGG and MG may be potentially useful as either anti-browning or anti-melanogenic agents in foods and cosmetics.

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Fractionation of Anticarcinogenic Enzyme Inducer(s) from Roasted Perilla (볶은 들깨박으로부터 암예방효소계 활성성분의 분획)

  • Hong, Eun-Young;Kang, Hee-Jung;Suh, Myung-Ja;Nam, Young-Jung;Kwon, Chong-Suk;Kim, Jong-Sang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.2
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    • pp.193-197
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    • 1997
  • Elevation of the activities of phase 2 enzymes such as quinone reductase(QR) provides protection against several types of neoplasia. In this study, we performed partial purification of QR inducer(s) from roasted and defatted perilla meal by solvent fractionation and thin layer chromatography. Cellular QR induction was most notable in chloroform fraction of roasted perilla extract, compared with other solvent fractions. QR inducer(s) was partially purified by TLC, with 0.8 of $R_f$ value in n-butanol : n-propanol : 2N-ammonium hydroxide(10 : 60 : 30). AHH-inducing activity in TLC fractions isolated from methanol extracts of roasted perilla comigrated with QR-inducing fraction, suggesting that QR and AHH are induced by the same compound. TLC fractions shown strong QR-inducing activity also had a potent antioxidative activity, suggesting that cellular QR enzyme is induced by antioxidant(s) present in roasted perilla.

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Effect of Citrus aurantium var amara on weight change in mice

  • Sarker, Satyajit Dey;Habibi, Bohlol;Sharifi, Tohid;Asnaashari, Solmaz;Nahar, Lutfun;Delazar, AndAbbas
    • Advances in Traditional Medicine
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    • v.8 no.3
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    • pp.222-227
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    • 2008
  • Citrus aurantium var. amara L., commonly known as 'bitter orange' or 'sour orange', of the family Rutaceae, has traditionally been used in the treatment of various ailments, and it possesses different types of pharmacological properties. As a part of our on-going studies on the plantsfrom the Iranian flora, the extract of C. aurantium var. amara has been studied for its weight lossproperties using the mice model. While the Sep-Pak fraction, 20% methanol (MeOH) in water, of the hydro-methanolic extract of the peels of C. aurantium var. amara fruits, when injectedintraperitoneal (i.p.) at a dose of 10 mg/kg, significantly decreased the level of weight gain of the mice in comparison with control the group (P < 0.01), the Sep-Pak fraction 80% MeOH in water decreased the initial weight of mice by 0.44% in six weeks. The administration of the total extract(10 and 20 mg/kg, i.p.), and the Sep-Pak fractions, 40% and 60% MeOH in water (10 mg/kg, i.p.)did not show any significant change of weight of the test mice. Of the two active fractions, the80% MeOH in water fraction did not show any noticeable adverse effects on mice, and was therefore analysed by reversed-phase preparative high performance liquid chromatography resulting in the isolation and identification of four major components, two coumarins, meranzin hydrate (1) and bergamottin (2), and two flavonoids, xanthomicrol 5,4'-di-methyl ether (tangeritin, 3) and hymenoxin 5,7-di-methyl ether (nobiletin, 4).

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.

Antimicrobial Effects of Ethanol Extracts from Korean and Indonesian Plants (국내 및 인도네시아산 식물의 에탄올 추출물의 항균효과)

  • Kim, Moo-Sung;Lee, Dong-Cheol;Hong, Jong-Eun;Chang, Ih-Seop;Cho, Hong-Yon;Kwon, Yong-Kwan;Kim, Hee-Yun
    • Korean Journal of Food Science and Technology
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    • v.32 no.4
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    • pp.949-958
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    • 2000
  • Antimicrobial effects of 150 kinds of Korean and 82 kinds of Indonesian plants were investigated to develope natural food preservatives. Extracts of the plants with 70% ethanol were tested their antimicrobial effects against several food spoilage microorganisms, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans and Aspergillus niger. Seventeen kinds of Korean and eighteen kinds of Indonesian plants were found relatively effective, of which Myristica fragrans and Melaleuca leucadendra were the most effective, respectively. The major fractions of the two plant extracts showing antimicrobial activity were further purified by solvent fractionation, silicagel column chromatography and preparative HPLC. The purified substances were identified as limonene and caprylic acid in M. fragrans, and ${\alpha}-terpineol$ in M. leucadendra, respectively.

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Study on the Components of the Anti-oxidant Activity of Panax Ginseng (인삼(人蔘)의 항산화(抗酸化) 성분(成分)에 관(關)한 연구(硏究))

  • Kim, Man-Wook;Choi, Kang-Ju;Cho, Yung-Hyun;Hong, Soon-Keun
    • Applied Biological Chemistry
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    • v.23 no.3
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    • pp.173-177
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    • 1980
  • The anti-oxidant components in Panax ginseng, which were known to have antiaging effect, were fractionated by various solvents and then isolated by the preparative thin layer chromatography. A developing system with silicagel G using toluene-chloroform-actene (20 : 13 : 13) as mobile phase was applied. The components were investigated the anti-oxidant activity by ${\alpha}$, ${\alpha}'-diphenyl-{\beta}-picrylhydrazyl$ (DPPH) at 517nm. Two effective components were obtained from red ginseng and were indophenol reducing substances.

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