• Title/Summary/Keyword: chromatography

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Improved Separation of Organic Explosives by Modified Micellar Electrokinetic Capillary Chromatography (Modified Micellar Electrokinetic Capillary Chromatography에 의한 폭약 성분의 분리능 향상)

  • Park, Sung-Woo;Yang, Young-Geun;Hong, Sungwook;Kim, Taek-Jae
    • Analytical Science and Technology
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    • v.10 no.5
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    • pp.325-331
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    • 1997
  • Among various CE separation methods, micellar electrokinetic capillary chromatography(MECC) method using sodium dodecylsulfate(SDS) provides rapid and accurate separation of organic explosive constituents with easy. The running buffer was composed with 2.5 mM borate and 25mM SDS(pH 8.5). Addition of 1M urea and 10% organic modifiers (acetonitrile, methanol and ethanol) improves the resolution of adjacent explosive constituents. When 15 explosive constituents were developed in MECC, most constituents were separated successively while RDX/TNB and DNN/DEP were not, and detection limits of separated compounds are in range of 1 to 4 ppm.

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Cyclodextrin Glucanotransferase from Bacillus stearothermophilus:Purification by Affinity Chromatography and Its Properties (Bacillus stearothermophilus가 생산하는 Cyclodextrin Glucanotransferase: Affinity Chromatography를 이용한 정제 및 성질)

  • 안중훈;황진봉;김승호
    • Microbiology and Biotechnology Letters
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    • v.18 no.6
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    • pp.585-590
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    • 1990
  • The cyclodextrin glucanotransferase (CGTase) was purified from the culture broth of Bacillus stearothermophilus by ammonium sulfate precipitation and affinity chromatography. The specific activity of the CGTase increased by about 31-fold from 111.5 U/mg protein to 3445.0 Ulmg protein. The SDSPAGE indicated that the purified CGTase was homogeneous and the molecular weight of the purified CGTase was about 78,000. The optimum pH and temperature was 6.0 and $60^{\circ}C$, respectively. This enzyme was stable from pH 5.5-10.0. The enzyme retained its full activity at the incubation temperature up to $60^{\circ}C$ and calcium ion increased the thermal stability. The isoelectric point was about 4.8.

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Comparison of the Exopeptidase Activity of Fractions from Crude Extracts of Octopus Octopus vulgaris Cuvier Hepatopancreas Using Different Fractionation Methods

  • Kim, Min Ji;Kim, Hyeon Jeong;Kim, Ki Hyun;Heu, Min Soo;Kim, Jin-Soo
    • Fisheries and Aquatic Sciences
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    • v.17 no.2
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    • pp.181-187
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    • 2014
  • This study was performed to identify the optimum fractionation method and conditions to obtain exopeptidase-active fractions from octopus hepatopancreas (HP) crude extracts (CEs) using four techniques: solid ammonium sulfate fractionation, polyethylene glycol (PEG) fractionation, anion exchange chromatography, and gel filtration chromatography. The fractions with the highest total activity toward L-leucine-p-nitroanilide (Leu-pNA) were fraction IV from the ammonium sulfate and PEG fractionation, and fraction II in ion exchange and gel filtration chromatography. The total exoprotease activity of these fractions was highest in fraction IV (4,050.20 U) of ammonium sulfate fractionation, followed by fraction II (3,600.28 U) from gel filtration chromatography, fraction IV (2,861.30 U) from PEG fractionation, and fraction II (2,576.28 U) from ion exchange chromatography. These results suggest that ammonium sulfate fractionation using 60-80% ammonium sulfate was the most efficient method for separating the exoprotease active fractions from CEs of octopus HP.

Purification and Application of Earthworm /alpha-Galactosidase by Affinity Chromatography (Galactomannan 이용에 관한 연구 : Affinity Chromatography에 의한 지렁이 유래 /alpha-Galactosidase의 정제 및 응용법)

  • 박귀근;정규훈;소림수행
    • Microbiology and Biotechnology Letters
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    • v.27 no.4
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    • pp.298-303
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    • 1999
  • An $\alpha$-D-galactosidase ($\alpha$-D-galactoside galactohydrolase, EC 3. 2. 1. 22) from earthworm was purified by affinity chromatography using N-$\varepsilon$-aminocaproyl-$\alpha$-D-galactopyranosylamine coupled to sepharose and its properties were examined. The specific activity of the purified enzyme, tested with p-nitrophenyl-$\alpha$-D-galactopyranoside as substrate, was 314 units/mg protein, representing an 122-fold purification of the original crude extract. The final preparation obtained from by Sephadex G-25 chromatography showed a single band on SDS-polyacrylamide gel electrophoresis. The molecular weight was determined to be 48,000 by SDS-polyacrylamide gel electrophoresis. The purified galactosidase was showed maximum activity at pH 4.5 and 4$0^{\circ}C$, and was stable in the pH and temperature ranges from 4.0 to 5.5 and 30 to 5$0^{\circ}C$, respectively. The enzyme activity was inhibited by Zn2+, Hg2+ and Co2+. When the purified $\alpha$-galactosidase treated to guar gum for 6 hour, gel-promoting property was increased. It was clear that enzymatic elimination of galactose from guar gum by purified $\alpha$-galactosidase would lead to a significant increase in gelation ability.

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Preparation of Amorphous Adefovir dipivoxil using Reverse Phase Column Chromatography and Solid Dispersion Method (역상컬럼 및 고체분산체를 이용한 무정형 아데포비어디피복실의 제조)

  • Yoon, Myeong-Sik;Oh, Da-Won;Maeng, Hyo-Chan;Hong, Hye-Suk;Park, Mi-Kyung;Lee, Yong-Tack;Lee, Si-Beum;Cho, Il-Hwan;Moon, Byoung-Seok
    • YAKHAK HOEJI
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    • v.54 no.4
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    • pp.316-321
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    • 2010
  • Adefovir dipivoxil which was originally developed by Gilead Sciences has been used as treatments of HIV and HBV, especially a therapeutics for HBeAg positive and negative chronic patients. We developed highly efficient purification method using reverse phase column chromatography for mass production and a stable amorphous Adefovir dipivoxil using solid dispersion method. Reverse phase column chromatography led to highly pure product, more than 99.7% by HPLC and can be used for mass production compared with normal column chromatography. Solid dispersion method containing watersoluble polymer and Isomalt showed improved stability of amorphous Adefovir dipivoxil against heat and moisture.

Separation of Burnup Monitors in Spent Nuclear Fuel Samples by Liquid Chromatography

  • Joe, Kih-Soo;Jeon, Young-Shin;Kim, Jung-Suck;Han, Sun-Ho;Kim, Jong-Gu;Kim, Won-Ho
    • Bulletin of the Korean Chemical Society
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    • v.26 no.4
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    • pp.569-574
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    • 2005
  • A coupled column liquid chromatography system was applied for the separation of the burnup monitors in spent nuclear fuel sample solutions. A reversed phase column was studied for the adsorption behavior of uranyl ions using alpha-hydroxyisobutyric acid as an eluent and used for the separation of plutonium and uranium. A cation exchange column prepared by coating 1-eicosylsulfate onto the reversed phase column was used for the separation of the lanthanides. In addition, retention of Np was checked with the reversed phase column and cation exchange column, respectively, according to the oxidation states to observe the interference effect for the separation of burnup monitors. This chromatography system showed a great reduction in separation time compared to a conventional anion exchange method. A good agreement from the burnup data was obtained between for this method and a conventional anion exchange method to within 1% of a difference for the spent nuclear fuel samples of about 40 GWD/MTU.

Identification and Characterization of Homoharringtonine from Cephalotaxus koreana

  • Kim, Byung-Sik;Kim, Jin-Hyun
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.566-569
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    • 2005
  • A novel purification method was developed for producing homoharringtonine from Cephalotaxus koreana, to guarantee high purity and yield. Our simple, efficient procedure for isolating and purifying homoharringtonine from C. koreanabiomass consisted of solvent extraction, synthetic adsorbent treatment, low-pressure chromatography, followed by high performance liquid chromatography (HPLC). The use of active clay treatment and silica gel low-pressure chromatography in the pre-purification process allowed for the rapid, efficient separation of homoharringtonine from interfering compounds and dramatically increased the yield and purity of crude homoharringtonine for high-performance liquid chromatography (HPLC) purification steps compared with alternative processes. Homoharringtonine could be obtained simply with high yield and purity from biomass using this purification method, while minimizing solvent use and the scale and complexity of HPLC operations for homoharringtonine purification. Purified homoharringtonine was identified and characterized.

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Isolation of Herbicidal Compounds from Pulsatilla koreana Roots (백두옹(Pulsatilia koreana Nakai) 뿌리로부터 제초활성물질의 분리)

  • 정형진
    • Korean Journal of Plant Resources
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    • v.9 no.1
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    • pp.47-54
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    • 1996
  • To search herbicidal compounds in Pulsatilla koreana Nakai, methanol extract of P. koreana roots was purified by sequences of XAD-7 column chromatography, silica gel adsorption column chromatography, silica gel flash column chromatography, preparative layer chromatography and preparative high performance liquid chromatography(Prep, HPLC).The final Prep. HPLC gave two herbicidally-active fractions. These fractions treatment at 100ppm inhibited the root length of Echinochloa crus-galli seedlings by 48% and 60% as compared with the control, respectively. Components in the two active fractions were analyzed by GC-MS Spectrometry. These compounds, which were isolated from P. koreana roots, were identified as several fatty esters, hydrocarbons, squalene, evidonol, and a diazepin analogue.

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The Use of the Online Two-dimensional Liquid Chromatography Coupled with a Universal Detector for the Screening of Non-volatile Potential Migrants in Food Packaging Materials (식품포장재내 비휘발성 잠재 이행물질들의 스크리닝을 위한 이차원크로마토 그래피와 범용검출기의 이용)

  • Yoon, Chan-Suk;Lee, Keun-Taik
    • KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
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    • v.16 no.1
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    • pp.9-18
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    • 2010
  • For screening test of the non-volatile compounds which migrate from food packaging materials into foodstuffs, the traditional high performance liquid chromatography (HPLC) systems suffer from the lack of universal detector with high sensitivity and universality and high efficiency HPLC separation column which provides complete separation of complex mixtures into all individual substances. In this work, the use possibility of online two-dimensional liquid chromatography (2D-LC) system coupled with a charged aerosol detector (CAD), a universal detector, was reviewed. 2D-LC system permits to improve peak capacity and resolving power for complex mixtures. Charged aerosol detector (CAD) offers a new feasibility for detection of any non-volatile compounds with high sensitivity and constant response factor in a calibration range. The combination of size exclusion chromatography (SEC) and normal phase HPLC (NP-HPLC) is most frequently used for the separation of the natural and synthetic polymers which are mainly used as raw materials for the manufacture of food packaging materials. However, there is no commercial software available for data acquisition and handling and therefore the quantification in 2D-LC analysis is still rare.

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A Study of Glutathione S-transferase Inhibitors obtained from Allium cepa var. cepa Extract (양파 추출물에서의 글루타티온 전달효소 활성 저해제에 관한 연구)

  • Lee, Kwang-Soo;Park, Kyung-Sook
    • The Korean Journal of Food And Nutrition
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    • v.26 no.4
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    • pp.725-730
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    • 2013
  • In this study, an ethanol extract being obtained from Allium cepa var. cepa examins the inhibitory effects on the glutathione S-transferase and the separation had been done by silica-gel column chromatography using various eluents, such as ethyl acetate, methanol, and 50% methanol. A volume of column fraction was 50ml and evaporation has been performde by the rotary evaporator under reduced pressure. Each fraction is being examined by thin layer chromatography and the UV spectrum at 365 nm was used to investigate separation patterns of spots on thin layer chromatography. When the eluent was changed, the spot patterns showed another different pattern on thin layer chromatography, so on. Fractions showing similar pattern are combined and eventually, three fractions are obtained. Each fraction is testified to examine the inhibition effects on glutathione S-transferase. All of these showed inhibition effects on glutathione S-transferase. The GC-MS shows that each fraction contains more than 2 compounds.