• Title/Summary/Keyword: 크로마토그라피

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Purification and Characterization of Lysyl Oxidase from Fetal Bovine Aorta in the presence of protease inhibitors -Evidence against polymorphism- (소(牛) 태아(胎兒) 대동맥(大動脈)으로부터 단백 분해효소 억제제 존제 하에 Lysyl Oxidase의 순수분리(純棒分離) -다양성(多形性)에 대(對)한 반론(反論)-)

  • Han, Song
    • Journal of Oral Medicine and Pain
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    • v.25 no.1
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    • pp.29-39
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    • 2000
  • Lysyl Oxidase from fetal bovine aorta was purified to homogenity using extraction, Sephacryl S200HR chromatography, Hydropore AX ion-exchange high performance liquid column chromatography, Cibacron blue affinity chromatography, and Sephacryl S-300 HR chromatography in the presence of protease inhibitor. The purified enzyme was active toward lathyritic collagen as well as elastin and was sensitive to aminonitriles such as BAPN. Upon Sephacryl S-300 HR chromatography, the enzyme was eluted as a peak with a $K_{av}$ value of 0.45 (65% of $V_t$ ) and it eluted from high performance liquid ion-exchange column (Hydropore |AX) at single position (ionic strength, I = 0.1~0.15). Once purified, it showed one band upon SDS-PAGE. It migrated to a band the mobility of which corresponded to a Mr of 33,500 upon reduction while it migrated to a 24,500 Mr position under the non-reducing condition. In constrast to other reports, it is concluded that fetal bovine aorta contains only one type of lysy oxidase.

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Determination of Polycyclic Aromatic Hydrocarbons in Chinmey Soot Extract by High-Resolution Gas Chromatography (고분리능 기체크로마토그라피에 의한 굴뚝재추출물안의 다환방향족 탄화수소의 정량에 관한 연구)

  • 서영화;이광우;허귀석
    • Journal of Korean Society for Atmospheric Environment
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    • v.4 no.2
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    • pp.11-19
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    • 1988
  • 대기오염물질 속에 함유되어 있는 다환방향족 탄화수소 (PAHs)는 분자식 구조가 비슷한 수십개의 이성질체가 여러 종류의 유기화합물과 혼합되어 존재한다. 본 연구에서는 환경오염 시료중의 PAHs를 분석하는데 있어서 분석방법과 결과를 비교하기 위하여 지침이 될 수 있는 환경표준 기준물을 개발할 목적으로 굴뚝 안쪽벽에서 긁어 채취한 검댕을 시료로 선책하여 액체/액체 용매 추출방법에 의해 PAHs 분류부분을 얻었다. Phenanthrene 이외의 30여종의 PAHs 화합물을 가스크로마토그라피의 머무른시간과 가스크로마토그라피/질량분석기에 의하여 분리, 판명하였고 5종의 주 PAHs 화합물을 정확하게 정량 분석하였다. 정량분석 결과의 신뢰도, 정확도, 정밀도는 미국 NBS의 표준기준물 1647을 분석하여 검정치와 비교함으로써 평가하였다.

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크로마토그라피를 이용한 당에스테르의 분리정제

  • Jo, Jeong-Yeon;Kim, Jae-Hun;Yu, In-Sang;Yu, Hyeon-Hui
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.630-631
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    • 2000
  • Sugar esters are produced by transesterification of fatty acids and sugars in organic solvent media. Chromatographic separation of sugar esters and fatty acids was investigated. $C_{18}$ column($3.9{\times}300mm$) was used in the chromatography experiment. The effect of flow rate, column temperature, and sample loading on the charateristics of chromatogram was studied. Sugar esters and fatty acids tested in this study was successfully separated with a high resolution.

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Development of Chromatographic Downstream Processing for The Purification of Monoclonal Antibody from Ascites Fluid: Part 1. Tandem Use of Hydroxylapatite Chromatography and Gel Permeation Chromatography (복수로부터의 단세포군 항체의 정제를 위한 크로마토그래프 분리 정제 시스템의 개발 - 1. 히드록실 아파타이트 크로마토그라프와 겔 여과 크로마토그라프로 이루어진 2단계 연속공정 사용 -)

  • Ahn, I.S.;Park, C.Y.
    • Microbiology and Biotechnology Letters
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    • v.17 no.1
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    • pp.19-23
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    • 1989
  • A sequential system composed of hydroxylapatite chromatography and gel permeation chromatography was developed to purify the IgM type monoclonal antibody against the colon cancer cell SC-1 from the ascitic fluid of mice injected with the murine hybridoma CH07E02. In the hydroxylapatite chromatographic step the band dilution could be reduced by controlling the gradient and flow rate of the eluent, the sodium phospate buffer, the optimum values for these variables being 5.82$\times$10$^{-3}$M/cm and 0.2$m\ell/\textrm{cm}^2$/min, respectively. A degree of purity better than 99.99% as judged from silverstaining of the SDS-PAGE bands, was obtained by adding the gel permeation chromatographic step in tandem.

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Characteristics of Bitter Peptides from a Cheese and a Soybean Paste (치즈 및 된장에서의 쓴 맛 펩타이드 특성)

  • Kim, Soo-Ho;Lee, Hyong-Joo
    • Korean Journal of Food Science and Technology
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    • v.17 no.4
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    • pp.276-282
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    • 1985
  • To characterize bitter peptides in fermented protein foods, peptides were extracted with 2:1 (v/v) chloroform-methand from various samples and separated into fractions I, II, and III by Sephadex G-25 gel chromatography. Amino acid compositions of Mozzarella cheese, soybean paste, and each fraction from the two samples were analyzed to calculate the average hydrophobicity. All the solvent extracts of the food samples had strong bitter taste, although the original samples did not taste bitter. The yield of solvent extraction ranged from 0.08 to 62.50% of total nigrogen of food samples. The average hydrophobicity calculated from the amino acid composition of Mozzarella cheese was 1376 cal/mole, solvent extract 1,623 cal/mole, gel chromatography traction I, 1,797 cal/mole, fraction II, 2,454 cal/mole, and fraction III, 1,559 cal/mole. In the case of soybean paste, the average hydrophobicity of original sample, solvent extract, gel chromatography fraction I, II, and III wre 1,229, 1,654, 1,900,998 cal/mole, respectively. The important amino acids in bitter peptides were leucine, 2016, phenylalanine, proline, and voline.

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Evaluation of vegetable oil rancidity by headspace gas chromatographic analysis (Headspace 가스크로마토그라피를 이용한 식용유의 산패도 측정)

  • Chun, Ho-Nam;Kim, Ze-Uook
    • Applied Biological Chemistry
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    • v.35 no.1
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    • pp.36-41
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    • 1992
  • Several commercial cottonseed, corn and rapeseed oils were stored at $60^{\circ}C\;and\;70^{\circ}C$ with daily exposure of fluorescent light for 12 hours and evaluated their rancidity by headspace gas chromatographic analysis of pentanal and hexanal. The data of gas chromatographic analysis was compared with organoleptic flavor evaluation. For headspace gas chromatographic analysis, the volatile compounds were recovered by porous polymer trap and flushed into a fused silica capillary column at $250^{\circ}C$. Twenty-three GC peaks were identified on the basis of relative retention time of reference compounds and gas chromatography-mass spectrometry. The results showed that the contents of pentanal and hexanal were linearly increased during storage. A very simple linear relationship was found between organoleptic flavor scores and amounts of two volatile compounds with very high correlation coefficient. This results suggested the possible implication of pentanal and hexanal as an quality index for rancidity evaluation of cottonseed, corn and rapeseed oils.

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A Gas Chromatographic Detector using Glow Discharge (글로우 방전을 이용한 기체크로마토그라피 검출기)

  • Han Chong Soo;Song, Seung Ho
    • Journal of the Korean Chemical Society
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    • v.35 no.4
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    • pp.350-354
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    • 1991
  • Characteristics of a gas chromatographic detector using glow discharge as the ionization source was studied in helium flow. Discharge current greater than 10$_6$ A was observed from the electric field 400 V/mm for the electrode distance 1 mm. The discharge current of 0.1~0.3 mA could be used for the detection of organic compounds. Discharge current was almost constant for the helium flow rate greater than 10 ml/min, but the discharge was easily disappeared by an injection of a small amount of organic compound in the flow rate of 0~30 ml/min. From the decrement of the discharge current depend on several compounds, it was suggested that the sensitivity of the glow discharge ionization chromatographic detector is strongly influenced by the molecular weight of the compounds.

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Properties of Ascorbate-Oxidizing Enzyme Purified from Pleurotus ostreatus (Pleurotus ostreatus에서 분리한 아스콜빈산 산화효소의 특성)

  • 황윤엽;김연란;강사욱
    • Korean Journal of Microbiology
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    • v.26 no.4
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    • pp.324-331
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    • 1988
  • Ascorbate oxidizing enzyme from the crude extract of Pleurotus ostreatus was purified by ammonium sulfate precipitation, preparative polyacrylamide gel electrophoresis, DEAE Sepharose CL-6B ion exchange chromatography and Sephadex G-150 gel filtration chromatography. The molecular weight of the enzyme estimated by Sephadex G-150 gel filtration chromatography was 140,000 and that of its subunit by SDS-polyacrylamide gel electrophoresis 66,000. The optimum pH for the maximum activity of the enzyme was 5.2 and the isoelectric point of the enzyme was 6.0 Km values for L-ascorbic acid and D-isoascorbic acid were both 2.2.$\mu$M, which indicates that the enzyme has the asme affinity towards both substrates.

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