• Title/Summary/Keyword: NMR analysis

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Vitis amurensis Ruprecht root inhibited ${\alpha}$-melanocyte stimulating hormone-induced melanogenesis in B16F10 cells

  • Jin, Kyong-Suk;Oh, You Na;Hyun, Sook Kyung;Kwon, Hyun Ju;Kim, Byung Woo
    • Nutrition Research and Practice
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    • v.8 no.5
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    • pp.509-515
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    • 2014
  • BACKGROUND/OBJECTIVES: The root of Vitis amurensis Ruprecht, a sort of wild-growing grape, has been used in oriental medicine for treatment of skin ailments; however, its dermatological activity is not sufficiently understood. The aim of this study was to investigate tyrosinase inhibitory and anti-melanogenic activities of V. amurensis Ruprecht root methanol extract (VARM) in B16F10 mouse melanoma cells and to attempt to isolate and identify the active compound issued from VARM. MATERIALS/METHODS: Anti-melanogenic activity of VARM was analyzed in ${\alpha}$-melanocyte stimulating hormone (MSH)-stimulated B16F10 cells through evaluation of antioxidative activity as well as inhibited tyrosinase activity and melanin contents compared with those of kojic acid and arbutin. After anti-melanogenic analysis of VARM, serial fractionation, nuclear magnetic resonance (NMR), and thin layer chromatorgraphy (TLC) were applied for identification of active compounds contained in VARM. RESULTS: VARM significantly inhibited oxidative stress and tyrosinase activity and attenuated ${\alpha}$-MSH-induced melanin production in B16F10 cells. For isolation of active compounds, VARM was fractionated using a series of organic solvents, including dichloromethane ($CH_2Cl_2$), ethyl acetate (EtOAc), and n-butanol (n-BuOH). Among fractions showing anti-melanogenic activity, the CH2Cl2 fraction induced the most potent attenuation of melanogenesis without cytotoxicity and the major compound in the $CH_2Cl_2$ fraction was identified as betulinic acid. Betulinic acid isolated from the $CH_2Cl_2$ fraction of VARM significantly attenuated ${\alpha}$-MSH-induced melanogenesis in a dose dependent manner, which was stronger than that of arbutin used as a positive control. CONCLUSIONS: These results indicate that VARM inhibits oxidative stress, tyrosinase activity, and ${\alpha}$-MSH-induced melanogenesis in B16F10 cells, due primarily to the active compound, betulinic acid, in the $CH_2Cl_2$ fraction.

Effects of Microbacterium laevaniformans Levans Molecular Weight on Cytotoxicity

  • Oh, Im-Kyung;Yoo, Sang-Ho;Bae, In-Young;Cha, Jae-Ho;Lee, Hyeon-Gyu
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.985-990
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    • 2004
  • Levans produced from Microbacterium laevaniformans were isolated, characterized, and fractionated by molecular weight. TLC, HPLC, and GC-MS analyses of the exopolysaccharide showed that it was a fructan-type polymer and was composed of (2,6)- and (2,1)-glycosidic linkages. $^{13}C$-NMR analysis proved that the polysaccharide was mainly a $\beta$-(2,6)-linked levan-type polysaccharide. To investigate the cytotoxicity of the acetone-precipitated levan fractions such as M1, M2, and M3, HepG2, P388D1, U937, SNU-1, and SNUC2A cell lines were screened. Among the cell lines tested, the cytotoxicity of M1- M3 fractions were detected from only SNU-1 and HepG2 cells at the dosage level of $100-800\mu\textrm{g}ml$. The M2 fraction M_r$, 80,000) at 400 $mu{g/ml}$ had the greatest cell growth inhibition (84.6%) on SNU-1, while the M1 $(M_r$, 50,000) at $800\mu\textrm{g}ml$ showed the greatest (46.32%) on HepG2. To obtain more uniform M_r$ fractions of levan, the levan was further fractionated from S1 $(M_r$ 1,000,000) to S5 $(M_r$ 10,000) using gel permeation chromatography. Again, the S1-S5 fractions had strong cytotoxicity on SNU-1 and HepG2 cell lines. The greatest inhibition effects of S4 $(M_r$ 80,000) on SNU-1 and S5 $(M_r$ 10,000) on HepG2 were shown to be 49.5% and 73.0%, respectively. The cytotoxicity of the levan fractions was more effective on SNU-1 than on HepG2. Although the relationship between the Mw and the cytotoxicity was not clear, smaller $M_r$, fractions of levan showed greater growth inhibition effect on the cancer cell lines in general. Therefore, it was indicated that a specific Mw class of levan is responsible for the effective cytotoxicity.

Antigenotoxicity of Ginseng Petroleum Ether Extract and its Action Mechanism (인삼 지용성성분인 유전독성억제효과와 작용기전)

  • 허문영
    • Journal of Food Hygiene and Safety
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    • v.13 no.3
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    • pp.243-251
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    • 1998
  • Panax ginseng C.A. Meyer has been extensively used in the traditional oriental medicine as a restorative, tonic and prophylatic agent. Petroleum ether extract of panax ginseng C.A. Meyer (GPE) and its several fractions (PI-P5) were tested for the evaluation of antigenotoxicity against N-methyl-N-nitrosourea (MNU) and benzo(a)pyrene [B(a)P]-induced micronucleated reticulocytes in mouse peripheral blood. GPE and P2 showed more significant anticlastogenicity than other fractions did. To elucidate the anticlastogenic action mechanism of GPE and P2 against B(a)P, the alteration of B(a)P metabolism was studied. GPE and P2 inhibited B(a)P metabolism in the presence of 8-9 mix and decreased B(a)P-DNA binding in calf thymus DNA with 8-9 mix. They also decreased [$^3H$] MNU induced DNA binding and methylation to 7-methyl guanine and $O^{6}-methyl$ guanine adducts in calf thymus DNA by RPLC analysis. These results suggest that the anticlastogenicity of GPE and P2 on the B(a)P or MNU-induced clastogenicity is due to decrease of DNA binding with B(a)P or MNU, the inhibition of metabolism with B(a)P and the inhibition of methylation in DNA. Therefore, GPE and P2 may be useful chemopreventive agents of alkylating agent like MNU and secondary carcinogen like B(a)P.

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An Antifungal Subatance, 2,4-Diacetylphloroglucinol Produced from Antagonistic Bacterium Pseudo-monas fluorescens 2112 Against Phytophthora capsici (Phytophthora capsici를 길항하는 Pseudononas fluorescens 2112가 생산하는 항진균 항생물질 2,4-diacetylphloroglucinol)

  • 이은탁;김상달
    • Microbiology and Biotechnology Letters
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    • v.29 no.1
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    • pp.37-42
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    • 2001
  • An antifungal substance was purified from culture broth of Pseudomonas flulorescens 2112 that showed a broad-spectrum antagonistic activity against various phytopathogenic fungi including capsici. The substance was identified as 2,4-diacetylphloro-glucinol basd on NMR analysis. The 2,4-diacetylphloroglcinol showed antibiotic activity in broad acidic range from pH 1.0 to pH 9.0. About 83% of initial activity was remained after incubation for 30min ar $60^{\circ}C$, however, the activity was dropped up to 50% after 30 min incubation in $80^{\circ}C$. When the nucleotides of P. capsici treated with 2,4-diacetylphloroglucinol were labeled with[$^{3}$ H]-Adenin, the newly synthesized and radioactive-labeled RNA was significantly reduced than those of untreated P. capsici. indicating that the 2,4-diacetylphloroglucinol inhibits RNA synthesis.

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Counter anion effects in anion exchange membrane-fabricated non-aqueous vanadium redox flow battery

  • Son, Pyeong Soo;Oh, Min-Seok;Ye, Jun-Hee;Choi, Seong-Ho
    • Analytical Science and Technology
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    • v.28 no.5
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    • pp.341-346
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    • 2015
  • In order to understand the counter anionic effects in a non-aqueous vanadium redox flow battery (VRFB), we synthesized four types of electrolyte salts (1-ethyltriethamine tertafluoroborate, [E-TEDA]+[BF4], 1-ethyltriethamine hexafluorophosphate, [E-TEDA]+[PF6], 1-butyltriethylamine tertafluoroborate, [B-TEDA]+[BF4], and 1-buthyltriethamine hexafluorophosphate [B-TEDA]+[PF6]) by counter anion exchange reaction after the SN2 reaction. We confirmed the successful synthesis of the electrolyte salts [E-TEDA]+[Br] and [B-TEDA]+[Br] via 1H-NMR spectroscopy and GC-mass analysis before the counter anion exchange reaction. The electric potential of the vanadium acetylacetonate, V(acac)3, as an energy storage chemical was shown to be 2.2 V in the acetonitrile solvent with each of the [E-TEDA]+[BF4], [E-TEDA]+[PF6], [B-TEDA]+[BF4], and [B-TEDA]+[PF6] electrolyte salts. In a non-aqueous VRFB with a commercial Neosepta AFN membrane, the maximum voltages reached 1.0 V and 1.5 V under a fixed current value of 0.1 mA in acetonitrile with the [E-TEDA]+[BF4] and [E-TEDA]+[PF6] electrolyte salts, respectively. The maximum voltage was 0.8 V and 1.1 V under a fixed current value of 0.1 mA in acetonitrile with the [B-TEDA]+[BF4] and [B-TEDA]+[PF6] electrolyte salts, respectively. From these results, we concluded that in the non-aqueous VRFB more of the [PF6] counter anion than the [BF4] counter anion was transported onto the commercial Neosepta AFN anion exchange membrane.

Induction of Apoptotic Cell Death in Human Jurkat T Cells by a Chlorophyll Derivative (Cp-D) Isolated from Actinidia arguta Planchon

  • Park, Youn-Hee;Chun, En-Mi;Bae, Myung-Ae;Seu, Young-Bae;Song, Kyung-Sik;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.27-34
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    • 2000
  • The chloroform and methanol (2;1, v/v) extract from an edible plant, Actinidia arguta Planchon, appeared to possess antitumor activity against human leukemias Jurkat T and U937 cells through inducing apoptosis. The substance in the solvent extract was purified by silica gel column chromatography, preparative TLC, and Sephadex LH-20 column chromatography. Characteristics of the substance analyzed by UV scanning analysis, $^1H$ and $^{13}C$ NMR spectra suggested that the substance belongs to the chlorophyll derivatives-like group. The $IC_{50}$ value of the chlorophyll derivative (Cp-D) determined by MTT assay was $15\mu\textrm{g}/ml$ for Jurkat, $10\mu\textrm{g}/ml$ for U937, and $11.4\mu\textrm{g}/ml$ for HL-60m and was more toxic to these leukemias than to solid tumors or normal fibroblast. In order to elucidate cellular mechanisms underlying the cytotoxicity, the effect of the Cp-D on Jurkat T cells was investigated. When cells were treated with the Cp-D at a concentration of $15\mu\textrm{g}/ml$, [3H]thymidine incorporation declined rapidly and wa undetectable in 1h. However, no significant changes were made in the cell cycle distribution of the cells by 24h. The sub-Gl peak representing apoptotic cells began to be detectable in 36h, at which time apoptotic DNA fragmentation was also detected on agarose gel electrophoresis, demonstrating that the cytotoxic effect of the Cp-D is attributable to the induced apoptosis. Under the same conditions, although the protein level of cyclin-dependent kinases such as cdc4, csk6, cdk2, and cdc2 was not significantly changed until 24h, the kinase activity of all c안 rapidly declined and reached a minimum level within 1-6h and then recovered to the initial level by 12h and sustained until 24h. These results suggest that inactivation of cdks at an inappropriate time during the cell cycle progression in jurkat T cells following a treatment with the Cp-D leads to induction of apoptotic cell death.

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Chemical Constituents of Domestic Quercus spp. Barks (국내산 참나무속 수종 수피의 추출성분)

  • Kim, Jin-Kyu;Kwon, Dong-Joo;Lim, Soon-Sung;Bae, Young-Soo
    • Journal of the Korean Wood Science and Technology
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    • v.38 no.4
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    • pp.359-374
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    • 2010
  • This study was carried out to investigate the chemotaxonomical correlation and chemical constituents of domestic Quercus spp. barks. The barks of Q. mongolica, Q. aliena, Q. serrata, Q. acutissima, Q. dentata, and Q. variabilis were collected in the experimental forest of Kangwon National University. The combined extracts were successively fractionated with n-hexane, methylene chloride and ethyl acetate using a separation funnel. A portion of the ethyl acetate and H2O soluble materials of each species were chromatographed on a Sephadex LH-20 column using various aqueous MeOH and EtOH-hexane as washing solvents. Spectrometric analysis such as NMR and MS, including TLC, were performed to characterize the structures of the isolated compounds. Ellagic acid (0.03 g), (+)-catechin (4.59 g), taxifolin (3.35 g), and glucodistylin (20.52 g) were isolated from Q. mongolica bark. Gallic acid (0.18 g), (+)-catechin (8.52 g), (+)-gallocatechin (0.09 g), taxifolin (0.54 g), and glucodistylin (3.28 g) were characterized from Q. acutissima bark. Gallic acid (0.38 g), ellagic acid (0.11 g), (+)-catechin (2.01 g), (+)-gallocatechin (0.12 g), and glucodistylin (0.39 g) were identified from Q. dentata bark. Ellagic acid (1.51 g), (+)-catechin (21.91 g), and glucodistylin (3.91 g) were purified from Q. aliena bark. Ellagic acid (0.84 g), (+)-catechin (0.82 g), taxifolin (4.02 g), and glucodistylin (21.50) were isolated from Q. serrata bark. Gallic acid (0.24 g), caffeic acid (0.05 g), (+)-catechin (0.32 g), and glucodistylin (0.65 g) were purified from Q. variabilis bark. (+)-Catechin and glucodistylin were isolated from all the barks. Glucodistylin can be a taxonomic index on Quercus spp.

Synthesis and Characterization of a Di-$\mu$-oxo-bridged Molybdeum(V) Complexes (두 개 산소가교형 몰리브덴(V) 착물의 합성과 그 성질에 관한 연구)

  • Doh, Gil Myung;Kim, Ill Chool;Choi, Bo Yong
    • Journal of the Korean Chemical Society
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    • v.39 no.3
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    • pp.198-203
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    • 1995
  • The Mo(V) $di-\mu-oxo$ type $(Mo_2O_4(H_2O)_2L)$ complexes $(L:\;C_3H_7CH(SCH_2COOH)_2,\;C_6H_5CH(SCH_2COOH)_2,\;CH_3OC_6H_4CH(SCH_2COOH)_2,\;C_5H_{10}C(SCH_2COOH)_2,\;C_3H_7C(CH_3)(SCH_2COOH)_2,\;C_3H_7CH(SCH_2CH_2COOH)_2,\;C_6H_5CH(SCH_2CH_2COOH)_2)$ have been prepared by the reaction of $[Mo_2O_4(H_2O)_6]^{2+}$ with a series of dithiodicarboxy ligands. These complexes are completed by two terminal oxygens arranged trans to one another and each ligand forms a chelate type between two molybdenum. In $Mo_2O_4(H_2O)_2L$, two $H_2O$ coordinated at trans site of terminal oxygens. The prepared complexes have been characterized by elemental analysis, infrared spectra, electronic spectra, and nuclear magnetic resonance spectra. In the potential range -0.00 V to -1.00 V at a scan rate of 20 $mVs^{-1}$, a cathodic peak at -0.50∼-0.58 V (vs. SCE) and an anodic peak at -0.40∼-0.43 V (vs. SCE) have been observed in aquous solution. The ratio of the cathodic to anodic current ($I_{pc}/I_{pa}$) is almost 1, we infer that redox is reversible reaction.

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Organotitanium Chemistry II. Solvolytic Reaction of Dichlorodiacetatotitanium (IV) with Various Alcohols (유기-티탄화학 (제2보). Dichlorodiacetatotitanium (IV) 여러 알콜과의 가용매반응)

  • Hoo Sung Lee;Young Sun Un;Sohn Youn Soo;Choi Q. Won
    • Journal of the Korean Chemical Society
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    • v.17 no.3
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    • pp.174-181
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    • 1973
  • Dichlorodiacetatotitanium(IV) dissolves in alcohols with chemical reaction. Such solvolytic reaction of $TiCl_2(OAc)_2$with various alcohols has been studied by means of solution NMR spectroscopy and chemical analysis of the isolated products. The reaction of $TiCl_2(OAc)_2$ with primary alcohols has shown to be a quantitative two-step ligand substitution process as shown in the following: $TiCl_2(OAc)_2+ROH{\to}TiCl_2(OAc)_2(OR)+AcOH$ $TiCl_2(OAc)_2(OR)+ROH{\to}TiCl_2(OAc)_2+AcOH$The molecular form initially soluble in organic solvents has been found to be the monosubstituted species $TiCl_2(OAc)(OR)$. Alcoholysis with t-butyl alcohol has shown remarkable differences. When the mole ratio of t-butyl alcohol to $TiCl_2(OAc)_2$ is less than 1/2, the following reaction is dominant. $TiCl_2(OAc)_2+t-ButOH{\to}TiCl_2(OAc)_2+t-ButCl$However, at higher mole ratio another substitution process resembling the first step reaction with primary alcohols is competitively accompanied. The reaction with t-butyl alcohol also differs from that with primary alcohols in that only one either of the two chloro-or acetato-ligands in $TiCl_2(OAc)_2$ is subjected to substitution.

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Metal Complexes of Ambidentate Ligand(Ⅳ). Nickel(Ⅱ) and Palladium(Ⅱ) Complexes of bis(isonitrosoacetylacetone)diimine Derivatives (Ambidentate 리간드의 금속착물 (제 4 보). Bis(isonitrosoacetylacetone)diimine 유도체를 리간드로 하는 니켈(Ⅱ) 및 팔라듐(Ⅱ) 착물)

  • Man-Ho Lee;Seon-Ho Lee
    • Journal of the Korean Chemical Society
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    • v.32 no.5
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    • pp.428-435
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    • 1988
  • Some nickel(Ⅱ) and palladium(Ⅱ) complexes of the ambidentate ligands derived from condensation of the isonitrosoacetylacetone and various diamines, $Ni(IAA)_2-en$, $Ni(IAA)_2-pn$, $Ni(IAA)_2-tn$, $Pd(IAA)_2-en$, PdCl(IAA)-pn, and $Pd(IAA)_2$-tn, where (IAA)$_2$-en, $(IAA)_2$-tn, and (IAA)-pn represent N,N'-enthylenbis(isonitrosoacetylacetone imine), N,N'-propylenebis(isonitrosoacetylacetone imine), N,N'-trimethylenebis(isonitrosoacetylacetone imine) and N-(2-aminopropyl)isonitrosoacetylacetone imine, respectively, have been prepared. The nickel(Ⅱ) and palladium(Ⅱ) complexes were characterized on the bases of the elemental analysis, IR, NMR, and electronic spectra. It is suggested that a isonitroso group of (IAA)$_2$-en or (IAA)$_2$-tn coordinates to the metal ion through the nitrogen atom to form five-membered ring, while the other isonitroso group of (IAA)$_2$-en or (IAA)$_2$-tn coordinates to the metal ion through the oxygen atom to form six membered ring in square-planar complexes of Ni(IAA)$_2$-tn and Pd(IAA)$_2$-en. And two isonitroso groups of (IAA)$_2$-en, (IAA)$_2$-pn, or (IAA)$_2$-tn coordinate to the metal ion through the nitrogen atom to form five-membered rings in square-planar complexes of Ni(IAA)$_2$-en, Ni(IAA)$_2$-pn, and Pd(IAA)$_2$-tn. On the other hand, square-planar PdCl(IAA)-pn is formed by the reaction of propylenediamine with a isonitrosoacetylacetone in the presence of palladium(Ⅱ)ion.

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