• Title/Summary/Keyword: Silica gel 60

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Isolation and Characterization of Elastase Inhibitor from Areca catechu (빈랑으로부터 Elastase 저해물질의 분리 및 특성조사)

  • 조중제;이건국;조병기;최정도
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.26 no.1
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    • pp.163-186
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    • 2000
  • We have previously screened 150 medicinal plants on the inhibition of elastase and found a significant inhibitory effects of the extracts of Areca catechu L. on the aging and inflammation against the skin tissues. To isolate and identify the compounds having biological activity, we was further purified by each of the solvent fractions, silica gel column chromatography, preparative TLC and reversed-Phase HPLC. Peak in HPLC, which coincided with the inhibitory activity against elastase, was identified as Phenolic substance using various colorimetric methods, UV, and IR. $IC_{50}$/ values of phenolic substance purified from Areca catechu were 26.9 $\mu\textrm{g}$/$m\ell$ for porcine pancreatic elastase (PPE) and 60.8 $\mu\textrm{g}$/$m\ell$ for human neutrophil elastase (HNE). This Phenolic substance showed more potent activity than those of reference compounds, oleanolic acid (76.5 $\mu\textrm{g}$/$m\ell$ for PPE, 219.2 $\mu\textrm{g}$/$m\ell$ for HNE) and ursolic acid (31.0 $\mu\textrm{g}$/$m\ell$ for PPE, 118.6 $\mu\textrm{g}$/$m\ell$ for HNE). According to the Lineweaver-Burk Plots, the inhibition against both PPE and HNE by this phenolic substance was competitive with substrate. Phenolic substance from Areca catechu exhibited high free radical scavenging effect ($SC_{50}$/ : 6 $\mu\textrm{g}$/$m\ell$) and inhibited effectively hyaluronidase activity ($IC_{50}$/: 210 $\mu\textrm{g}$/$m\ell$). These results suggest that the Phenolic substance Purified from Areca catechu showed anti-aging effect by protecting connective tissue proteins.

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Differential Effects of Resveratrol and its Oligomers Isolated from Seeds of Paeonia lactiflora (Peony) on Proliferation of MCF-7 and ROS 17/2.8 Cells

  • Kim, Hyo-Jin;Lee, Won-Jung;Park, Yun-Hee;Cho, Sung-Hee;Park, Sang-Won
    • Preventive Nutrition and Food Science
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    • v.8 no.4
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    • pp.356-364
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    • 2003
  • A methanol extract from seeds of Paeonia lactiflora (Paeoniaceae, peony) was found to possess different antiproliferative activities against four different human cancer cell lines: Hela, MCF-7, HepG2 and HT-29. Furthermore, five different methanol (20, 40, 60, 80 and 100 % MeOH) fractions obtained by fractionation of the methanol extract of the seeds on a Diaion HP-20 column exhibited differential antiproliferative effects against the above four cancer cell lines. Among five fractions, the 60 % MeOH fraction showed relatively lower antiproliferative activity on MCF-7 estrogen-sensitive breast cancer cell than the other cancer cell lines. Systematic separation of 60% the MeOH fraction by silica gel and Sephadex LH-20 columns led to the isolation of four known stilbenes, trans-resveratrol (1), trans-(+)- $\varepsilon$ -viniferin (2), gnetin H (3) and suffruticosol B (4). The four stilbenes (1∼4) exerted differential biphasic effects on cell proliferation of MCF-7 cells in a similar manner as genistein, a soybean isoflavone used as a positive reference, in the concentration range from 1.0 to 200 $\mu$M. Three stilbenes (1 ∼ 3) weakly stimulated the proliferation of MCF -7 cells at doses below 10 JIM. However, strong antiproliferative effects on MCF-7 cell were exerted by extract 1 at a dose of 200 JIM, and by 2 and 3 at doses above 25 $\mu$M. In contrast, 4 inhibited the proliferation of MCF-7 cell at a dose below 25 $\mu$M, but stimulated cell proliferation at concentrations of 50 and 100 $\mu$M. All four stilbenes (1∼4) stimulated the proliferation of ROS 17/2.8 osteoblast-like cells in the range of 10$^{-10}$ ∼10$^{-1}$ $\mu$M. Compound 1 exhibited especially potent proliferative activity, although its activity was weaker than that of genistein. Additionally, three resveratrol oligomers (2∼4) also exhibited concentration-dependently moderate proliferative activity, but less than that of 1. These results suggest that resveratrol, and its dimer and trimers from the seeds of Paeonia lactiflora may act as a phytoestrogen, but in a somewhat different manner from that of genistein.

The activity of antioxidants and suppression of cancer cell proliferation in extracts of Orostachys japonicus A. Berger (바위솔 추출물의 항산화활성 및 암세포 증식억제)

  • Kim, Chung-Hyun;Park, Jae-Ho;Lim, Jong-Kuk;Lee, Kon-Joo;Chung, Gyu-Young;Jeong, Hyung-Jin
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.1
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    • pp.31-39
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    • 2003
  • The antioxidative activities and biological properties in the EtOAc extracts and purified extracts of Orostachys japonicus were measured by assay of DPPH, xanthine/xanthine oxidase and mammalian cells(2-12). Scavenging of DPPH radical and inhibition of xanthine/xanthine oxidase of EtOAc extracts were showed the highest activity in the arable land and in September. The S-4 fraction showed the highest activity among the silica-gel column chromatography fractions. LH-4 fraction showed higher activity than the other fractionsins in assay of DPPH and xanthine/xanthine oxidase. Fatty acids and phenolic compounds were identified by GC/MS and main compounds were 1,2,3-benzenetriol, alpha-androsta-7,14-diene in LH-4 fraction. The activities of POD and SOD in samples havested on different habitats were high such as arable land> intermountain> seashore. That of POD and SOD in crude extracts of late stage were higher than early stage. Isozyme bands of crude extracts samplinged in all habitats and all growing stages showed two bands and the signal intensity showed strongly according to passage of growing stage. The purified extracts of LH-4 fraction showed excellent inhibition effect in proliferation of HL-60 cells and markedly suppressed colony formation in mouse fibroblast cells. Dose response between partially purified extracts(400ppm) and negative control did not produced statistically significant reduction in colony formation.

Colored coating of SiO2-TiO2-MxOy(M = Cu, Co, Cr) thin films by the sol-gel process (졸-겔법에 의한 SiO2-TiO2-MxOy(M=Cu, CO, Cr)계 박막의 제조 및 색상에 관한 연구)

  • Kim, Sangmoon;Lim, Yongmu;Hwang, Kyuseog
    • Journal of Korean Ophthalmic Optics Society
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    • v.3 no.1
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    • pp.229-235
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    • 1998
  • This paper reports the preparation and characterization of colored coatings of $SiO_2-TiO_2-M_xO_y$ (M = Co, Cr or Cu). Films of different compositions ranging from a molar content of transition metals of 5% to 20% have been prepared on soda-lime-silica slide glasses by the sol-gel process. The films have been characterized by a photospectroscopy. The color and reflectance of the films was expressed in Lab color chart and on spectra plot. 'L' as lightness and all reflectance decreased with increase of the content of transition metals. The coating of Co, Cu and Cr cotaining system showed light blue, green and lemon-yellowish color, respectively.

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Coating and Characterization of Al2O3-CoO Thin Films by the sol-gel Process (졸-겔법을 이용한 Al2O3-CoO계 박막의 제조와 특성에 관한 연구)

  • Shim, Moonsik;Lim, Yongmu
    • Journal of Korean Ophthalmic Optics Society
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    • v.4 no.2
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    • pp.123-128
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    • 1999
  • This paper reports the preparation and characterization of colored coatings of $Al_2O_3$-CoO. Films of 25mol% CoO doped $Al_2O_3$, have been prepared on soda-lime-silica slide glasses by the sol-gel process from Al-alkoxide and Co-nitrate. The films have been characterized by a photospectroscopy and hardness tester. The color, spectral reflectance and spectral transmittance of the films was expressed in Lab color chart and on spectra plot. Microhardness of the films increased with increasing of the heating temperature. Transmittance and reflectance of the films decreased with increase of the heating temperature and coating times. The coating films showed various light-yellow, deep-yellow, greenish-yellow color as a function of the coating times and heating temperature.

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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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Studies on antimicrobial agent produced by lactobacillus acidophilus (Lactobacillus acidophilus가 생산한 항균물질(抗菌物質)에 관한 연구(硏究))

  • Kim, Dong-shin
    • Korean Journal of Veterinary Research
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    • v.24 no.2
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    • pp.149-162
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    • 1984
  • The research was conducted(1) to confirm the agent(s) responsible for the antimicrobial activity contained in the fermented tomato juice with L. acidophilus(2) to extract and purify the antimicrobial agent(s)(3) to find the biological, physical and chemical properties of the agent(s). The following results were obtained and summarized as followings; 1. The agent responsible for the inhibitory activity was confirmed by both well assay method using fermented tomato juice with L. acidophilus and turbidimetric technique using the cell-free filtrate or neutralized filtrate of tomato acidohilus culture and found exerted antimicrobial agent other than lactic acid. 2. The procedures of purification : The isolation and purification of antimicrobial agent from the lyophilized acidophilus tomato culture were carried out by (1) methanol extraction (2) acetone extraction, (3) Sephadex G-50 gel filtration (4) paper chromatography and (5) thin layer chromatography. 3. The biological, physical and chemical properties of antimicrobial agent: The biological, physical, chemical properties of the purified antimicrobial agent were: (1) The antimicrobial activity was strong against test organisms; Bacillus subtilis(ATCC 6633), Escheichia coli(ATCC 25922), Staphylococcus aureus(ATCC 167), Pseudomonas fluorescens(KFCC 32394), Proteus vulgaris and Shigella dysenteriae. (2) The pH value of the agent was 2.0 and the inhibitory activity was lost when it was neutralized at 7.0 of pH and the agent was heat stable at $121^{\circ}C$ for 60 minutes. (3) The ultraviolet light absorption spectra of methanol-acetone extract and TLC fraction exhibited a maximum absorption at 260nm and 224nm respectively. (4) The most purified agent from TLC plate increased about 130-fold in activity. (5) The agent isolated from TLC plate was free from $H_2O_2$ or lactic acid. 4. Bioautographic assy: By means of bioautography of the agent on silica gel of TLC plate a strong inhibitory activity against B. subtilis was demonstrated. 5. Mass spectrometry: The agent obtained from TLC plate was analyzed by mass spectrometry which show the parent peak at m/e 264 suggesting the molecular weight of the compound and molecular group such as [$C_2H{^+}_4$], [CO], [CH=NH], [$C_3{H^}4_7$], [$\begin{array}{rcl}O\\{\parallel}\\CH_3-C\\\end{array}$], [$C_6-H{^+}_{11}$], [$C_5H{^+}_{11}$], [$\begin{array}{rcl}O\\{\parallel}\\C_5H_7-C^+\\\end{array}$] were suggested.

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Anti-cancer Activity of Styrax japonica Bark Extrats (때죽나무(Styrax japonica) 수피 추출물의 항암 활성)

  • Kwon, Oh-Woong;Kim, Woo-Jin;Lee, Hak-Ju
    • Journal of the Korean Wood Science and Technology
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    • v.42 no.1
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    • pp.68-77
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    • 2014
  • A compound has been isolated from the methanol extract of Styrax japonica bark using conventional chromatographic methods including silica gel chromatography, TLC and HPLC. The molecular formula of Styraxlignolide F analyzed by spectrometric analyses using FAB-MS, NMR was found to be $C_{27}H_{34}O_{11}Na$. The cytotoxicity of the styralignolide F was showed 15.2% in $1.0mg/m{\ell}$ on human kidney cell (HEK 293). As anticancer activity of $CH_2Cl_2$ fraction, over 60% of AGS and MCF-7 cells were inhibited in concentration of $1.0mg/m{\ell}$. In the results of anticancer test using quantification of Bcl-2, $CH_2Cl_2$ fraction showed lower Bcl-2 and p53 expression than those of styraxlignolide F and other fractions. In apoptosis of human lung carcunoma cancer cell (A549), $CH_2Cl_2$ fraction showed the highest inhibition rate (46.9%) and styralignolide F was the next (43.5%). The $CH_2Cl_2$ fraction showed higher anti-cancer activities than isolated substance (styraxlignolide F), probably due to the crude extract showing synergic effects by other components.

Isolation and Identification of Antioxidant Substances from the Stems of Butterbur (Petasites japonicus) (머위(Petasites japonicus) 엽병으로부터 항산화 물질의 분리 및 동정)

  • Kim, Min-Young;Yi, Jung-Hyun;Hwang, Yun-Yi;Song, Kyung-Sik;Jun, Mi-Ra
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.8
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    • pp.979-984
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    • 2008
  • The stems of P etasites japonicus were extracted with ethanol and then partitioned with hexane, chloroform, ethyl acetate, n-butanol and water, successively. The antioxidant potency of five crude fractions were determined using (1) 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, (2) thiobarbituric acid reactive substances (TBARS) assay in the linoleic acid model system, and (3) lipoxygenase inhibition assay. Among the crude fractions, the ethyl acetate fraction exhibited the most potent antioxidant effect. By activity-guided fractionation, compound PJ-4 was isolated from the ethyl acetate fraction through the repeated silica gel open column chromatography. The chemical structure of the isolated compound was determined as kaempferol by $^1H-$ and $^{13}C$-NMR analysis and its antioxidative capacity was further investigated. DPPH radical scavenging activity of the compound was 65.76% at the concentration of $100 \;{\mu}g/mL$. The inhibitory activity of the compound against lipid peroxidation and lipoxygenase exhibited 43.47% and 58.60%, respectively at the concentration of $100\;{\mu}g/mL$. The result suggests that the compound may serve as a useful natural antioxidant and furthermore indicates the possibility of developing the stems of Petasites japonicus as a natural antioxidant source.

INHIBITION OF BROWNING REACTIONS OCCURRING IN THE STORAGE OF DRIED OYSTER 2. Inhibitory Effect of Sodium Sulfite Treatment and the Addition of Antioxidants (건조굴 저장중의 갈변방지 2. 아황산소오다 처리 및 항산화제 효과)

  • CHOI Jin-Ho;LEE Kang-Ho;KIM Mu-Nam
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.10 no.1
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    • pp.17-22
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    • 1977
  • In the present paper, the effect of sodium sulfite treatment on tile inhibition of browning reactions occurring in the storage of dried oyster was tested and the supplementary effect of antioxidantsaddedwasalsomentioned. Dried oysters treated with sodium sulfite solutions as described in the previous paper(Lee and Choi, 1977) were stored in the bottles with silica gel bags at room temperature with or without the application of antioxidants. The ethanol solution of an antioxidant mixture(BHA, BHT, plus, synergists) was sprayed on the surface of cooked oyster before drying. The density of brown pigment was determined spectrophotometrically by measuring the absorbance at 420 and 440 nm of both fractions of pigment extract, namely chloroform-methanol and water soluble fractions, which represent the brown color developed by fat oxidation and Maillard reactions respectively. TBA value was also measured for the oxidative rancidity in oysters during the storage. It appeared from the results that the 0.5 M sodium sulfite-60minute treated samples showed better effect after 150 day storage at room temperature. Controlling tile pH of treating solutions, did not reveal so much different in inhibitory effect in the aspect of color but a more reduction of tyrosine and reducing sugar was resulted with acidic solution than with alkaline solution. The development of brown color in dried oyster seemed to be leaded rather by the oxidative rancidity of lipids than sugar-amino reactions particularly in a long-term storage since the browning of chloroform-methanol fraction progressed more rapidly than of water. soluble fraction. The application of antioxidant, therefore, could largely retard the browning of the product as appeared in the results that sodium sulfite treated oyster with addition of antioxidant kept the best color during the storage.

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