• Title/Summary/Keyword: 크로마토그래피

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Adsorption Characteristics of Liquid Chromatography with Preparative Packings (제조용 충전물을 사용한 액체 크로마토그래피의 흡착특성)

  • Choi, Yong Seok;Lee, Chong Ho;Row, Kyung Ho
    • Applied Chemistry for Engineering
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    • v.9 no.3
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    • pp.430-434
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    • 1998
  • Preparative HPLC (High-Performance Liquid Chromatography) is mainly used for separating useful component from biological samples. By reversed-phase HPLC packed with preparative packings ($15{\mu}m$), the adsorption characteristics with sample size were investigated. Sample was 5'-GMP, a flavor enhancer, and the composition of mobile phase was 20mM $KH_2PO_4$ solution:methanol (97:3 vol.%). From the experimental results, the effect of sample size on retention factor was negligible, but the peak was asymmetrical above $1{\mu}g$ of sample. In addition, the increase in sample size deteriorated the number of theoretical plates, and at small concentration, the number of theoretical plates was less because of large peak width. In the experimental condition, the adsorption isotherm of 5'-GMP was relatively well represented by Freundlich equation.

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Purification and Characterization of Xylanase from Bacillus sp. A-6 (Bacillus sp. A-6의 Xylanase의 정제와 특성)

  • Choi, Suk-Ho
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.147-152
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    • 2009
  • A xylanase was purified from the culture supernatant of Bacillus sp. A-6 by using ultrafiltration and ion exchange chromatography on the column of SP-Sepharose using 5 mM acetate buffer, pH 5.0. The xylanase was eluted from the column at the concentration less than 0.05 M NaCl. The eluted xylanase was shown to be a single protein band in SDS-PAGE. Zymogram analysis indicated that the protein band in SDS-PAGE had the enzyme activity to hydrolyze oat spelt xylan. The molecular weights of the xylanase were 15,000 based on SDS-PAGE and 14,100 based on gel filtration chromatography. Thin layer chromatography showed that the xylanase hydrolyzed oat spelt xylan into xylobiose and high-molecular-weight xylooligosaccharides. The relative activities of the heated xylanase decreased to 80% at $40^{\circ}C$ after 7 hr and less than 40% at $60^{\circ}C$ after 1 hr.

토양균에서 항생물질 및 효소억제제의 분리와 구조 연구

  • 구양모;이윤영;김경자;최응칠;김범태;주정호;이창훈
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.179-179
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    • 1994
  • 새로운 항생물질을 개발하기 위하여 토양으로부터 분리한 균주를 액체 및 고체배지에서 배양하여 여러 검정균에 대하여 종이디스크법으로 항균효력을 조사하였다. 그 결과 (+), G(-), fungi 등에 강한 항균 효력을 보인 토양균 SNUS 8810-43과 Mycobacterium, fungi에 항균력을 보인 토양균 SNUS 8810-129를 선택하여 각각의 배양액에서 항생물질을 분리하고, 분리한 항생물질의 구조를 규명하고자 하였다. 토양균 SNUS 8810-43의 배양액으로부터 항생물질을 분리하기 위하여 양이온 교환 수지 관 크로마토그래피와 셀룰로오스 관 크로마토그래피를 수행하여 시료 JJH-II-46-43을 얻었다. 시료 JJH-II-46-43의 IR, $^1$H-NMR, $^{13}$C-NMR, $^1$H-$^1$H COSY, $^1$H-$^{13}$C COSY, FAB-MS 스펙트럼을 얻어 분리한 항생물질의 구조를 분석하여 이 항생물질의 구조가 N-methylstreptothricin과 동일하다는 것을 확인하였다. Mycobacterium smegmatis에 강한 활성을 나타내는 물질을 토양균 SNUS 8810-129로 부터 분리하였다. 토양균 SNUS 8810-129를 배양한 V-8 아가판을 메탄올로 추출하여 이를 실리카겔 관 크로마토그래피와 preparative TLC로 시료 LCH-IV-17B, LCH-III-387을 얻었다. 시료LCH-IV-l7B, LCH-III-387의 $^1$H-NMR, $^{13}$C-NMR, FAB-MS, CI-MS, IR등의 스펙트럼을 얻어 분리한 항생물질의 구조를 분석하여 이 항생물질이 glycolipid계 항생물질이라는 것을 알았다. $^{13}$C-NMR 상의 자료와 화학적인 방법으로 구성당을 조사한 결과 이 항생물질을 이루고있는 당은 rhamnose 임을 알았다. 또 이 항생물질을 구성하는 지방산은 화학적인 방법과 MS 스펙트럼, $^{13}$C-NMR 스펙트럼으로부터 hydroxydecanoic acid인 것으로 확인되었다. 항생물질 LCH-III-387와 항생물질 LCH-IV-l7B는 각각 rhamnose를 1, 2개 포함하고 있는 것으로 확인되었다. 그리고 동일한 탄소수의 지방산을 가지고 있는 것으로 생각되었다. 이들 항생물질을 이루는 구성당과 지방산간의 정확한 연결및 구조, 생리활성에 관한 연구는 계속 수행중에 있다.

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Development of New Separation Technique, Modifier Composition Programming in Supercritical Fluid Chromatography (초임계 유체 크로마토그래피에서 새로운 분리방식인 변형제 조성 프로그래밍법 개발)

  • Kim, Hohyun;Pyo, Dongjin
    • Analytical Science and Technology
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    • v.10 no.5
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    • pp.350-356
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    • 1997
  • Supercritical Fluid Chromatography(SFC) has been developed as an analytical technique for the compounds that is difficult to analyze by conventional chromatography. Since supercritical fluid $CO_2$ is difficult to elute solutes with high polarity, modified supercritical $CO_2$, was used as a mobile phase. In conventional method, silica column which is saturated with modifier was used. However, with this method, we can not control the quantity of modifier. In this paper, we developed a new method which can control quantity of modifier mixed in supercritical fluid $CO_2$. The quantity of $H_2O$ mixed was measured with amperometric microsensor which was made by perflurosulfonate ionomer(PFSI) film. we have also obtained a good supercritical fluid chromatogram of PAH mixture by use of a modifier composition programming method.

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Enrichment of Lithium Isotopes by Cation Exchange Chromatography (양이온 교환 크로마토그래피에 의한 리튬 동위원소의 농축)

  • Kim, Dong Won;Kim, Chang Suck;Choi, Ki Young;Jeon, Young Shin;Jeong, Young Kyu;Park, Sung Up
    • Analytical Science and Technology
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    • v.7 no.2
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    • pp.201-204
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    • 1994
  • Cation exchange column chromatography of lithium was carried out to investigate the lithium isotope separation in aqueous ion exchange system. A Pyrex glass column of $50cm{\times}6mm$ inner radius with a water jacket was used as the separation column in experiment. Upon column chromatography using hydrochloric and succinic acid mixtures as an elunent, single separation factor, ${\alpha}$, 1.0068 was obtained. From the experiment, it was found that $^6Li$ was enriched in the resin phase and $^7Li$ in the solution phase.

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Purification of Glucose Oxidase by Affinity Chromatography and Its Characterization (친화성 크로마토그래피를 이용한 글루코오스 옥시다아제의 정제와 효소특성)

  • Ko Jung Hwan;Byun Si Myung
    • Journal of the Korean Chemical Society
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    • v.23 no.3
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    • pp.165-174
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    • 1979
  • A purification technique of glucose oxidase was developed. Using the gluconyl-${\omega}$-aminohexyl Sepharose affinity chromatography, it was partially purified 14.6 folds with 79.7% yield. With the combination of the affinity chromatography and Sepharose 6B gel filtration, the enzyme was purified 27.2 folds from the broth with 74.1% yield. The final purified preparation showed 90.83 U of glucose oxidase activity per mg of protein and a single band by 7% polyacrylamide gel electrophoresis. The absorption spectrum and substrate specificity of the enzyme were studied and the fianal preparation showed the optimal pH between 5.6 and 6.0, the optimal temperature at $40^{\circ}C$, $8.5{\times}10^{-3}M$ of $K_m$ for D-glucose, and 3.43 kcal/mole of the activation energy.

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Simulation of Preparation Protein Chromatography (제조용 단백질 크로마토그래피의 시뮬레이션)

  • 김인호;이선묵;황우성
    • KSBB Journal
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    • v.14 no.3
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    • pp.371-376
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    • 1999
  • Simulation of preparative protein chromatography becomes necessary for separation as well as optimal operation. A mathematical model describing the behavior of elution peaks in preparative protein chromatography for single and binary component separation was solved numerically using a PDEsolver Macsyma$^{\circledR}$(Macsyma Inc., Arlington, MA, U.S.A.). Band profiles were calculated with the equilibrium-dispersive model of chromatography. The effects of the sample volume, concentrations of solutes in the sample, flow velocity and column length on the band profile of the elution peaks are discussed. The results in this paper suggest the model simulation for the binary mixture can be extended to multicomponent separations.

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Studies on the Toxic Activity of Bacillus sphaericus Spo -D1216 Asporogenic Mutant (Bacillus sphaericus Asporogenic Mutant Spo - D1216의 독성에 관한 연구)

  • 복거중;김영한;이형환
    • Microbiology and Biotechnology Letters
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    • v.13 no.2
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    • pp.157-162
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    • 1985
  • The cell cultures and crude extracts of Bacillus sphaericus 1593 K-5 and its mutant Spo-Dl216 were respectively bioassayed against Culex pipiens var. pollens mosquito larvae. The B. sphaeriucs 1593 K-5 showed toxic activity against the larvae. LC$_{50}$ values (cells/$m\ell$) was 2.6$\times$10$^2$. Also the LC$_{50}$ ($\mu\textrm{g}$ Protein/$m\ell$) of the crude extract was 10.26. However, B. sphaericus Spo-Dl216 didn't show toxic activity against the larvae. The soluble cytoplasmic toxin in broken B. sphaeriucs 1593k-5 cells was partially purified by gel permeation chromatography and ion exchange chromatography. Among the fractions of the gel permeation chromatography only a single fraction was found to be toxic. LC$_{50}$ values ($\mu\textrm{g}$ protein/$m\ell$) of the active fraction was 0.182. The active fraction of the gel permeation was subjected to ion exchange chromatography. Only a single fraction showed toxic activity and its LC$_{50}$ values ($\mu\textrm{g}$ protein/$m\ell$) was 0.02..02.

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