• Title/Summary/Keyword: fast atom bombardment/MS

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Chemical Synthesis and Determination of Biological Activity of the Epidermal Growth Factor-Like Domain of Mouse Betacellulin

  • Shin, Song-Yub;Kang, Shin-Won;Ha, Jong-Myung
    • BMB Reports
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    • v.28 no.2
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    • pp.87-93
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    • 1995
  • To investigate the biological functions of the EGF-like domain of mouse betacellulin (BTC), mouse BTC(33-80), a 48-residue peptide corresponding to the EGF-like domain, was synthesized by stepwise solidphase methods using a 9-fluorenylmethoxycarbonyl (Fmoc) strategy. The homogeneity of synthetic mouse BTC(33-80) was confirmed by analytical reversed phase (RP)-HPLC, amimo acid analysis, and fast atom bombardment mass spectrometer (FAB-MS). Three disulfide bond pairings of synthetic mouse BTC(33-80) were established by amino acid analysis of cysteine-containing fragments derived from thermolytic digestion. These were consistent with the pairings of EGF and transforming growth factor ($TGF-{\alpha}$). The EGF-Iike domain of mouse BTC showed equipotent activity in both EGF-receptor binding on A-431 epidermoid carcinoma cells, and mitogenesis on NIH-3T3 fibroblast cells, as compared with authentic h-EGF. Results suggest that the EGF-Iike domain of BTC plays a significant role in mitogenic activity with an EGF-receptor mediated system.

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Rapid Separation and Indentification Method of Tea Catechins (녹차 중 카테킨류의 신속 분리 및 동정법)

  • Lee, Jeong-Hee;Lee, Yong-Moon;Moon, Dong-Cheul
    • Analytical Science and Technology
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    • v.5 no.3
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    • pp.333-338
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    • 1992
  • The tea tannins, epigallocatechin, epigallocatechin gallate, were successfully separated by a Sephadex LH-20 column by the acetone based gradient elution. Each fractions was collected by monitoring at 280nm. Purified fractions were directly characterized by fast atom bombardment mass spectrometry. Epigallocatechin and epigallocatechin gallate were identified and shown as low as 70% purity in the reversed phase column. This revised method is more advantageous than known methods in purity and rapidity.

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Isolation and Characterization of Bacillus Strains for Biological Control

  • Kim, Han-Soo;Park, Jiyong;Cho, Sung-Won;Park, Kee-Hyun;Lee, Gung-Pyo;Ban, Soo-Jung;Lee, Chang-Roo;Kim, Chung-Sun
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.196-201
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    • 2003
  • The object of this study was to characterize Bacillus strains GB-017 and GB-0356, which produce antifungal substances, especially for plant pathogens. In addition, this study was undertaken to characterize the culture conditions required for the production of antifungal substances and to document some of the properties of the antifungal substance produced by these soil-isolated strains. Strains GB-0365 and GB-017 were found to be bacillus-shaped, gram-positive and motile, and to inhibit Botrytis cineria, Fusarium sp., Pythium sp., and Rhizoctonia solani. Antagonistic activity was maintained up to pH 9.0, and the antifungal activity was stable to heat at 80$^{\circ}C$ for 1 h. Antifungal substances were separated and purified using ion exchange and adsorption columns including WK-I0(H$\^$+) (pH 7.0), HP20 column (pH 3.0) and IPA (pH 3.0). and IPA. Its UV absorption spectrum showed major peaks at 231 and 259 nm, corresponding to polyene and lactone. A fast atom bombardment mass spectrum (FAB MS) showed a highest peak at 441 m/z and major peaks at 192, 205, and 370 m/z.

Re-evaluation of physicochemical and NMR data of triol ginsenosides Re, Rf, Rg2, and 20-gluco-Rf from Panax ginseng roots

  • Cho, Jin-Gyeong;In, Seo-Ji;Jung, Ye-Jin;Cha, Byeong-Ju;Lee, Dae-Young;Kim, Yong-Bum;Yeom, Myeonghun;Baek, Nam-In
    • Journal of Ginseng Research
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    • v.38 no.2
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    • pp.116-122
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    • 2014
  • Ginseng roots were extracted with aqueous methanol, and extracts were suspended in water and extracted successively with ethyl acetate and n-butanol. Column chromatography using the n-butanol fraction yielded four purified triol ginseng saponins: the ginsenosides Re, Rf, Rg2, and 20-gluco-Rf. The physicochemical, spectroscopic, and chromatographic characteristics of the ginsenosides were measured and compared with reports from the literature. For spectroscopic analysis, two-dimensional nuclear magnetic resonance (NMR) methods such as $^1H$-$^1H$ correlation spectroscopy, nuclear Overhauser effect spectroscopy, heteronuclear single quantum correlation, and heteronuclear multiple bond connectivity were employed to identify exact peak assignments. Some peak assignments for previously published $^1H$-and $^{13}C$-NMR spectra were found to be inaccurate. This study reports the complete NMR assignment of 20-gluco-Rf for the first time.

Physico-Chemical Properties and Antimicrobial Activity of Pyocyanine Produced by Pseudomonase aeruginosa KLP-2 (Pseudomonas aeruginosa KLP-2가 생산한 Pyocyanine의 항균활성 및 생리화학적 성상)

  • 박은희;이상준;차인호
    • Journal of Life Science
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    • v.11 no.5
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    • pp.483-488
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    • 2001
  • The antimicrobial substance produced by Pseudomonas aeruginosa KLP-2 strain was purified and identified. The substance was identified as a pyocyanine by the fast atom bombardment mass(FAB-MS). In physic-chemical properties, the pyocyanine was dark blue needles, and was soluble in various organic solvents such as chlorogorm, methanol, ethanol and ethyl acetae. The pyocyanine possessed a ultraviolet absorbance spectrum in methanol, 0.1 M HCl, and chlorogorm. The maximum absorption peak of the pyocyanine showed at 318 mm in methanol. The molecular formula of the pyocyanine was determined to the $C_{13}$ H$_{10}$ N$_{2}$O and protonate molecular ion species (M+H)$^{+}$ was observed at m/z 211 by FAB-MS. The pyocyanine showed antimicrobial against Bacillus cereus, Bacillus subtilis, Micrococcus luteus, Rodococcus equi, Staphylococcus aureus, Streptococcus faecalis, E. col, Legionella pneumophila, Shigella flexneri Shigella boydii, shgella sonnei, NAG Vibrio cholerae, Vibrio parahaemolyticus, Vibro vulnificus, Yersinia enterocolitica, and Saccharomyces cerevisiae. However, Salmonella spp. Shigela dysenteriae, 3 strains of Pseudomonas aeruginosa, Klebsiela pneumoniae, and Aspergillus niger were resistant to the pyocyanine. The pyocyanine showed the highest antimicrobial activity aganist Legionella pneumophila based on the size of inhibition zone by the disk contained 0.5 $\mu\textrm{g}$ of the pyocyanine.e.

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Identification of NMR Data for ginsenoside Rg1 (Ginsenoside Rg1의 NMR 데이터 동정)

  • Lee, Dae-Young;Cho, Jin-Gyeong;Lee, Min-Kyung;Lee, Jae-Woong;Park, Hee-Jeong;Lee, Youn-Hyung;Yang, Deok-Chun;Baek, Nam-In
    • Journal of Ginseng Research
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    • v.32 no.4
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    • pp.291-299
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    • 2008
  • The fresh ginseng roots were extracted in aqueous methanol (MeOH), and the obtained extracts were partitioned using ethyl acetate (EtOA), n-butanol (n-BuOH), and water, successively. The repeated silica gel column chromatography for n-BuOH fraction afforded a purified ginsenoside $Rg_1$. The physico-chemical, spectroscopic and chromatographic data of ginsenoside $Rg_1$, such as crystallization characteristics, melting point, specific rotation, infrared spectrometry (IR) data, fast atom bombardment/mass spectrometry (FAB/MS) data, nuclear magnetic resonance (NMR) data, retention factor (Rf) in thin layer chromatography (TLC) experiment, and retention time (r.t.) in HPLC analysis, were measured and compared with those reported in literatures. Especially, the previous literatures reported different data for ginsenoside $Rg_1$ in the $^{1}H-$ and $^{13}C$-NMR experiments. This paper gives the exactly assigned NMR data through 2D-NMR experiments, such as $^{1}H-^{1}H$ correlation spectroscopy (COSY), hetero nuclear single quantum correlation (HSQC), and hetero nuclear multiple bond connectivity (HMBC).

Identification of Water Soluble Metabolites of Pentachlorophenol(PCP) in the Suspension Cultures of Soybean and Rice Cells;3. Identification of PCP Glucose conjugates (콩과 벼 현탁배양(懸濁培養) 중 PCP 수용성대사물(水溶性代謝物)의 동정(同定);3. PCP glucose conjugates의 동정(同定))

  • Kim, Pil-Je;Park, Chang-Kyu
    • Korean Journal of Environmental Agriculture
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    • v.15 no.2
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    • pp.167-178
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    • 1996
  • In order to identify PCP glucose conjugates transformed from PCP in soybean and rice cell suspension cultures, the purified metabolites were acetylated, purified twice by HPLC using a normal and a reversed phase column, and then subjected to fast atom bombardment(FAB) mass spectrometric analysis. As were the conjugates, their acetylated derivatives of the glucose conjugates formed at the early stage(48 hr) of metabolism were separated by HPLC into three fractions. FABMS analysis of each fraction revealed that, at least in two fractions, the locations of the spectral peaks were practically coincident with those deducible from the structures of pentachlorophenyl and tetrachlorophenyl ${\beta}-D-glucopyranosides$. Based on information obtained from mass spectral and chromatographic analysis of not only the water-soluble metabolites but also aglycones and glycone, it is concluded that PCP is primarily metabolized to glucose conjugates, which account for more than 50% recovery of the PCP-conveyed radioactivity from the water soluble metabolites : The conjugates are mainly made up of pentachlorophenyl ${\beta}-D-glucopyranoside$, tetrachlorophenyl ${\beta}-D-glucopyranosides$( probably 2 or more isomers), and 2-hydroxy-3,4,5,6-tetrachlorophenyl ${\beta}-D-glucopyranoside$.

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Identification of Maysin and Related Flavonid Analogues in Corn Silks (옥수수 수염에서 Maysin 및 유사물질의 동정)

  • Kim, Sun-Lim;Snook, Maurice E.;Kim, E-Hun;Park, Cheol-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.45 no.3
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    • pp.151-157
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    • 2000
  • This study was carried out to isolate and identify the maysin and related flavonoid analogues in corn silks. Silks were covered with silk bag to prevent pollination and were sampled at 3-5 days after silking. The silks were filled with 100% MeOH and stored at $0^{\circ}C$ until analysis. The MeOH extracts of corn silks were filtered and concentrated at 35-4$0^{\circ}C$. The ${CH}_2$${Cl}_2$ was added on the concentrated aqueous solution to remove the chlorophyll and lipids. The Cis open column (25mm$\times$54 cm) was washed and activated with serial treatment of 500$m\ell$ of 100% MeOH(twice)longrightarrow75% MeOH longrightarrow50% MeOHlongrightarrow30% MeOHlongrightarrow100% $H_2$O(2 times). The concentrated aqueous solution was applied to the $C_{18}$ column and washed with $H_2O$ several times to remove the sugars and water soluble pigments. Neochlorogenic acid, chlorogenic acid and 4-caffeoylquinic acid were eluted with 10% MeOH, and rhamosyl isoorientin was eluted with 30% MeOH, but maysin was eluted with 50% MeOH from the $C_18$ open column. Collected fractions were analyzed with HPLC by using revers-phase Ultras-phere $C_{18}$ column (4.6$\times$250mm, 5$\mu\textrm{m}$) and $H_2$O (10% MeOH containing 0.1% $H_3$${PO}_4$)/MeOH (100% MeOH containing 0.1% H$_3$PO$_4$) linear gradient from 20% to 90% MeOH for 35 minutes, a flow rate of 1 $m\ell$/min and detection at 340nm. The selected fractions were concentrated and applied to the silicic acid column. Maysin was eluted with 500$m\ell$ of 100% ethyl acetate from the silicic acid column for the first purification, and the purity of collected fractions was about 75%, but the purity from the second purification with the Cis column (1/2 $\times$ 43") was greater than 95%. FAB-MS spectral data was obtained with VG7O-VSEQ VG analytical fast atom bombardment mass (UK). $^1$H-NMR and $^{13}$ C-NMR data were obtained with Bruker DPX 400 MHz NMR spectrometers (German) in DMSO-d$_{6}$ at 400 and 100 MHz, respectively.vely.

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