• 제목/요약/키워드: chromatography

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

  • 박성우;양영근;홍성욱;김택제
    • 분석과학
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    • 제10권5호
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    • pp.325-331
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    • 1997
  • 계면활성제로 sodium dodecylsulfate(SDS)를 이용한 micellar electrokinetic capillary chromatography (MECC) 방법을 이용하여 유기 폭약 성분을 신속하고 정확하게 분석할 수 있는 방법을 연구하였다. 완충요액으로는 2.5mM borate와 25mM SDS(pH 8.5)를 사용하였고, 완충용액에 1M urea와 10% 유기 용매(acetonitrile, methanol 및 ethanol)를 첨가한 결과 분리능이 향상되는 현상을 관찰할 수 있다. 15종의 폭약 혹은 그 구성 성분을 MECC 방법으로 분리한 결과 RDX/TNB 및 DNN/DEP를 제외한 성분을 분리할 수 있었고, 검출한계는 1~4ppm으로 나타났다.

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

  • 안중훈;황진봉;김승호
    • 한국미생물·생명공학회지
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    • 제18권6호
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    • pp.585-590
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    • 1990
  • Bacillus stearothermophilus가 생산하는 cyclodextrin glucanotransferase (CGTase)를 ammonium sulfate 법과 affinity chromatography법에 의해 정제하였다. 이 CGTase의 비활성은 11.5U/mg protein에서 33445.0U/mg protein으로 증가하였다. 이 효소의 분자량을 측정하기 위해 SDS-PAGE를 실시한 결과 분자량은 약 7,800이었다. 이 효소의 최적 pH와 온도는 각각 6.0과 $60^{\circ}C$이었다. 이 효소는 pH5.5와 10.0에서 안정하였다. 이 효소에 calcium ion을 첨가하였더니 열안정성이 증가하였다. 이 효소의 등전점은 약 4.8 이었다.

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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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    • 제17권2호
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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.

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

  • 박귀근;정규훈;소림수행
    • 한국미생물·생명공학회지
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    • 제27권4호
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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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역상컬럼 및 고체분산체를 이용한 무정형 아데포비어디피복실의 제조 (Preparation of Amorphous Adefovir dipivoxil using Reverse Phase Column Chromatography and Solid Dispersion Method)

  • 윤명식;오다원;맹효찬;홍혜숙;박미경;이용택;이시범;조일환;문병석
    • 약학회지
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    • 제54권4호
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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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    • 제26권4호
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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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    • 한국생물공학회 2005년도 생물공학의 동향(XVII)
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    • pp.566-569
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    • 2005
  • 한국산 개비자나무로부터 새로운 개념의 homoharringtonine 분리 및 정제 공정을 개발하였다. 메탄올을 사용한 4회 반복 추출에 의해 biomass로부터 대부분(>99%)의 homoharringtonine을 회수 할 수 있었다. 흡착 공정에서는 활성 백토(active clay)를 사용하여 추출물에 포함되어 있는 식물유래 타르, 왁스 성분을 효과적으로 제거하였다. Silica gel low-pressure chromatography공정을 통해서 순도 52% 이상의 homoharringtonine을 얻었으며, HPLC 공정을 통해 고순도 및 고수율의 homharringtonine을 정제할 수 있었다. 정제된 homoharringtonine의 분자량 및 구조분석을 수행한 결과 표준물질(Homharringtonine)과 동일 물질임을 확인할 수 있었다.

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

  • 정형진
    • 한국자원식물학회지
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    • 제9권1호
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    • pp.47-54
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    • 1996
  • 할미꽃의 뿌리에 함유되어 있는 제초활성물질의 확인과 동정을 위하여 이들을 추출 정제하고 활성성분 및 제초활성을 조사하여 다음과 같은 결과를 얻었다. 1. 각종 칼럼크로마토그래피법과 Prep. HPLC의 순차적인 적용에 의해 제초활성물질이 포함된 할미꽃 뿌리의 메타놀추출물을 정제하여 2개의 활성분획을 획득하였다. 2. 최종적으로 얻어진 활성분획들을 100ppm의 농도로 조제하여 처리한 결과, 식용피 유근의 생장 억제율은 대조구에 비해 각각 48% 및 60%로 나타났다. 3. GC-MS Spectrometry에 의해 분리된 활성분획 중의 성분들을 기존자료들과 비교하여 수종의 지방산 에스테르, 2종의 탄화수소류, squalene, evidonol 및 1종의 diazepin 관련 화합물이 동정되었다.

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식품포장재내 비휘발성 잠재 이행물질들의 스크리닝을 위한 이차원크로마토 그래피와 범용검출기의 이용 (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)

  • 윤찬석;이근택
    • 한국포장학회지
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    • 제16권1호
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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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양파 추출물에서의 글루타티온 전달효소 활성 저해제에 관한 연구 (A Study of Glutathione S-transferase Inhibitors obtained from Allium cepa var. cepa Extract)

  • 이광수;박경숙
    • 한국식품영양학회지
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    • 제26권4호
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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.