• 제목/요약/키워드: column plot

검색결과 45건 처리시간 0.032초

Presence of Rhodanese in the Cytosolic Fraction of the Fruit Bat (Eidolon helvum) Liver

  • Agboola, Femi Kayode;Okonji, Raphael Emuebie
    • BMB Reports
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    • 제37권3호
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    • pp.275-281
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    • 2004
  • Rhodanese was isolated and purified from the cytosolic fraction of liver tissue homogenate of the fruit bat, Eidolon helvum, by using ammonium sulphate precipitation and CM-Sephadex C-50 ion exchange chromatography. The specific activity was increased 130-fold with a 53% recovery. The $K_m$ values for KCN and $Na_2S_2O_3$ as substrates were $13.5{\pm}2.2\;mM$ and $19.5{\pm}0.7\;mM$, respectively. The apparent molecular weight was estimated by gel filtration on a Sephadex G-100 column to be 36,000 Da. The optimal activity was found at a high pH (pH 9.0) and the temperature optimum was $35^{\circ}C$. An Arrhenius plot of the heat stability data consisted of two linear segments with a break occurring at $35^{\circ}C$. The apparent activation energy values from these slopes were 11.5 kcal/mol and 76.6 kcal/mol. Inhibition studies on the enzyme with a number of cations showed that $Mg^{2+}$, $Mn^{2+}$, $Ca^{2+}$, and $Co^{2+}$ did not affect the activity of the enzyme, but $Hg^{2+}$ and $Ba^{2+}$ inhibited the enzyme.

역상액체크로마토그래피에서 벤조산류의 머무름 (Retention of Benzoic Acids in RPLC)

  • 이재석;최범석
    • 분석과학
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    • 제13권3호
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    • pp.269-276
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    • 2000
  • 역상액체크로마토그래피에서 7종의 벤조산의 머무름에 대해서 연구하였다. 벤조산의 불감시간 측정은 무기염이 적당하였다. 로그 용량인자(log k')와 Hildebrand의 용해인자(${\delta}$)는 직선관계를 가지며 머무름 메카니즘이 소용매성 효과에 영향을 받는다. 용질의 전이 엔탈피는 온도증가 그리고 이동상에서 유기변형제의 비와 반비례 관계를 가진다. 이동상에서 물의 부피분율과 log k'의 관계로부터 얻은 기울기인 S값은 양의 값을 가진다. 자유에너지 변화는 유기변형제의 부피분율의 증가와 비례한다. 소수성지표 ${\varphi}_0$은 컬럼의 온도와 반비례한다.

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현호색의 Acetylcholinesterase 활성 저해 성분 및 그 작용기전 (An Acetylcholinesterase Inhibitor Isolated from Corydalis Tuber and Its Mode of Action)

  • 황세영;장영표;변순정;전미희;김영중
    • 생약학회지
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    • 제27권2호
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    • pp.91-95
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    • 1996
  • In the course of searching for acetylcholinesterase inhibitors from crude drugs, it was found that total MeOH extract of Corydalis Tuber showed significant inhibitory effect on acetylcholinesterase. To isolate acetylcholinesterase inhibitors from Corydalis Tuber, total MeOH extract of the the crude drug was subjected to activity guided fractionation. The MeOH extract was suspended in water and fractionated with methylene chloride and subjected to acid-base fractionation. Silica gel column chromatography of the basic fraction which showed significant inhibitory effect on acetylcholinesterase was carried out and 5 subfractions (1-5) were obtained. From subtraction 4, compound I was isolated. The structure of isolated compound I was identified by spectroscopic parameters of $^1H-NMR$, $^{13}C-NMR$, EI-MS and FAB-MS. The compound I was identified as berberine. It was found from the Lineweaver-Burk plot that berberine was a reversible and specific inhibitor of acetylcholinesterase having 90% inhibitory effect at the concentration of $2.5{\mu}M$.

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Parameter Estimation of Perillyl Alcohol in RP-HPLC by Moment Analysis

  • Row Kyung Ho;Lee Chong Ho;Kang Ji Hoon
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제7권1호
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    • pp.16-20
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    • 2002
  • Parameter estimations were made for the reversed-phase adsorption of perillyl alcohol (POH), a potent anti-cancer agent, on octadecylsilyl-silica gel (ODS). The average particle diameter of ODS was about $15\;{\mu}m$, and the particles were packed in the column $(3.9\;\times\;300mm)$. The mobile phase used was a mixture of acetonitrile and water, in which the acetonitrile ranged between 50 and $70\;(v/v\;\%)$. The first absolute moment and the second central moment were determined from the chromatographic elution curves by moment analysis. Experiments were carried out using POH solutions within the linear adsorption range. The fluid-to-particle mass transfer coefficient was estimated using the Wilson-Geankoplis equation. The axial dispersion coefficient and the intra particle diffusivity were determined from the slope and intercept of a plot of H vs $1/u_0$, respectively. The contributions of each mass-transfer step were axial dispersion, fluid-to-particle mass transfer, and intraparticle diffusion.

Ganoderma lucidum이 생산하는 Polygalacturonase의 정제 및 특성 (Purification and Properties of Polygalacturonase from Ganoderma lucidum)

  • 윤숙;김명곤;홍재식;김명숙
    • 한국균학회지
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    • 제22권4호
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    • pp.298-308
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    • 1994
  • Ganoderma lucidum이 생산하는 polygalacturonase의 유용활용 방안을 위한 효소의 특성을 연구하기 위하여 버섯배양물로부터 효소를 생산하고 정제하여 그 특성을 검토한 결과는 다음과 같다. Endo-polygalacturonase는 배양여액으로부터 ammonium sulfate 침전, Biogel P-100, DEAE-cellulose, Sehpadex G-150 column chromatography에 의하여 순차적으로 정제되었고, Sehpadex G-150 column chromatography에서 re-gel filtration까지 실시한 결과 specific activity가 892unit/mg protein로 배양여액보다도 약 56배의 정제도를 나타냈으며, exo-polygalacturonase는 ammonium sulfate침전, Biogel P-100, DEAE-cellulose column chromatography까지 부분정제한 결과 9.2배의 정제도를 나타냈다. SDS polyacrylamide gel에서 전기영동을 실시한 결과 단일 band를 나타냈으며, 분자량은 54,000 dalton이었다. Endo-polygalacturonase와 exo-polygalacturonase는 citrus pectin이나 pectic acid보다 apple pectin에 친화도가 높았으며, Lineweaver-Burk plot로부터 계산된 apple pectin에 대한 Km 치는 각각 1.44와 10.6 mg/ml이었다. 이 두 효소의 작용 최적 pH는 4.0이었으나 pH 안정성에서는 endo-polygalacturonase는 $pH\;4.0{\sim}6.0$에서, exo-polygalacturonase는 $pH\;3.5{\sim}5.5$ 범위에서 안정하였다. 효소반응 최적 온도는 endo-polygalacturonase는 $40^{\circ}C$ 이었으나 exo-polygalacturonase는 $60^{\circ}C$에서 최대의 활성을 나타냈고 열안정성도 exo-polygalacturonase가 endo-polygalacturonase보다 더 높은 온도에서도 안정하였다. 또한 endo-polygalacturonase는 $Ca^{++}$$Mn^{++}\;ion$에, exo-polygalacturonase에는 $Ca^{++}\;ion$에 의해 효소반응이 촉진되었다.

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Aspergillus niger가 생산(生産)하는 섬유소(纖維素) 분해효소(分解酵素)의 정제(精製) 및 특성(特性) (Purification and Characterization of Cellulolytic Enzymes from Aspergillus niger)

  • 박관화;오태광;신재두
    • Applied Biological Chemistry
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    • 제24권3호
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    • pp.186-193
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    • 1981
  • Aspergillus niger가 분비하는 섬유소분해효소(纖維素分解酵素)를 Sephadex G-150 column을 통과시켜서 ${\beta}-glucosidase$와 CM-cellulase을 분리(分離)하고 CM-cellulase는 다시 DEAE-Sephadex A-50 column을 통과시켜서 세개의 CM-cellulase F I, F II , F III를 분리하여 그 특성(特性)을 각기 조사 하였다. F I의 최적조건(最適條件)은 pH 4.0 및 $50^{\circ}C$였고 열불활성화특성(熱不活性化特性)은 Arrhenius plot에서 2개의 기울기가 다른 직선을 나타냈다. 즉 $60{\sim}70^{\circ}C$ 사이에서는 Z값이 $4^{\circ}C$ 이고 $70{\sim}98^{\circ}C$ 사이에서는 $383^{\circ}C$였다. F II는 pH 4.7 및 $50^{\circ}C$에서 최적조건(最適條件)을 보였고 Z값은 $8^{\circ}C$, F III는 pH 4.3 및 $60^{\circ}C$에서 높은 활성(活性)을 가졌고 Z값은 $10^{\circ}C$였다. 조(粗) CM-cellulase의 경우는 최적조건(最適條件)이 pH 4.3 및 $60^{\circ}C$로 F I , F II 및 F III와 비슷한 경향을 보였으나 Z값은 $21.5^{\circ}C$로 큰 차이(差異)를 보였다. 한펀 ${\beta}-glucosidase$는 정제효소(精製酵素) 및 조효소(粗酵素)가 다 같이 pH 4.7 및 $60^{\circ}C$에서 가장 높은 활성(活性)을 나타냈으며 Z값은 $7^{\circ}C$였다.

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Discrimination of Panax ginseng Roots Cultivated in Different Areas in Korea Using HPLC-ELSD and Principal Component Analysis

  • Lee, Dae-Young;Cho, Jin-Gyeong;Lee, Min-Kyung;Lee, Jae-Woong;Lee, Youn-Hyung;Yang, Deok-Chun;Baek, Nam-In
    • Journal of Ginseng Research
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    • 제35권1호
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    • pp.31-38
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    • 2011
  • In order to distinguish the cultivation area of Panax ginseng, principal component analysis (PCA) using quantitative and qualitative data acquired from HPLC was carried out. A new HPLC method coupled with evaporative light scattering detection (HPLC-ELSD) was developed for the simultaneous quantification of ten major ginsenosides, namely $Rh_1$, $Rg_2$, $Rg_3$, $Rg_1$, Rf, Re, Rd, $Rb_2$, Rc, and $Rb_1$ in the root of P. ginseng C. A. Meyer. Simultaneous separations of these ten ginsenosides were achieved on a carbohydrate analytical column. The mobile phase consisted of acetonitrile-water-isopropanol, and acetonitrile-water-isopropanol using a gradient elution. Distinct differences in qualitative and quantitative characteristics for ginsenosides were found between the ginseng roots produced in two different Korean cultivation areas, Ganghwa and Punggi. The ginsenoside profiles obtained via HPLC analysis were subjected to PCA. PCA score plots using two principal components (PCs) showed good separation for the ginseng roots cultivated in Ganghwa and Punggi. PC1 influenced the separation, capturing 43.6% of the variance, while PC2 affected differentiation, explaining 18.0% of the variance. The highest contribution components were ginsenoside $Rg_3$ for PC1 and ginsenoside Rf for PC2. Particularly, the PCA score plot for the small ginseng roots of six-year old, each of which was light than 147 g fresh weight, showed more distinct discrimination. PC1 influenced the separation between different sample sets, capturing 51.8% of the variance, while PC2 affected differentiation, also explaining 28.0% of the variance. The highest contribution component was ginsenoside Rf for PC1 and ginsenoside $Rg_2$ for PC2. In conclusion, the HPLC-ELSD method using a carbohydrate column allowed for the simultaneous quantification of ten major ginsenosides, and PCA analysis of the ginsenoside peaks shown on the HPLC chromatogram would be a very acceptable strategy for discrimination of the cultivation area of ginseng roots.

헤드스페이스 GC/MS를 이용한 폴리염화비닐/폴리염화비닐리덴 식품 포장 중 염화비닐/염화비닐리덴 잔류량 연구 (A Study on Residual Amounts of Vinyl Chloride/Vinylidene Chloride in Poly(vinyl chloride)/Poly(vinylidene chloride) Food Packaging using Headspace GC/MS)

  • 성준현;곽인신;박성관;김형일;임호수;이주영;변미연;김소희
    • 한국식품과학회지
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    • 제43권1호
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    • pp.6-11
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    • 2011
  • PVC 및 PVDC 재질의 식품포장에 잔류할 수 있는 원료물질인 VC 및 VDC를 PLOT Q 컬럼이 장착된 headspace-GC/MS를 이용하여 분석하였다. VC 및 VDC 각각에 대하여 m/z=61, 62, 64 및 m/z=61, 96, 98 이온을 선택 이온으로 하여 분석하였으며, 내부표준 물질로는 1-CP를 사용하였다. 분석법 검증결과 본 연구에서 도입한 분석법은 유럽연합의 재질규격 시험에 적합하였으며, 국내유통 PVC 및 PVDC 재질의 식품 포장 103 품목에 대한 분석 결과, PVDC 식품포장 1건에서 재질 중 VDC가 검출되었으나, 이는 유럽연합에서 설정하고 있는 재질규격 이내에 드는 미미한 수준이었으며, 나머지 모든 시료에서는 VC 및 VDC가 검출되지 않았다.

역상 액체 크로마토그래피에서 페놀 일치환체들의 머무름에 미치는 치환기들의 영향 (Effect of Substituted Groups on the Retention of Monosubstituted Phenols in Reversed-Phase Liquid Chromatography)

  • 김훈주;이인호;이대운
    • 대한화학회지
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    • 제38권8호
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    • pp.562-569
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    • 1994
  • $ C_{18}$ 및 Phenyl 컬럼에서 메탄올-물 및 아세토니트릴-물 혼합액을 이동상으로 하여 21종의 페놀일치환체들의 머무름을 조사하여 치환기가 머무름에 미치는 영향을 알아보았다. 아미노페놀을 제외한 모든 시료의 머무름은 Phenyl 컬럼보다 $ C_{18}$컬럼에서 더 컸으며, 두 이동상에 대해서는 물의 부피퍼센트가 같을때 메탄올계에서 더 컸다. 일반적으로 일치환 페놀류의 파라 이성질체는 메타 이성질체보다 빨리 용리되었고, 오르토와 메타 이성질체 사이에서는 용리순서가 치환기의 극성에 따라 다르게 나타났다. 페놀 일치환체들의 머무름은 시료와 미반응된 실란올과의 상호작용의 영향을 받는대 이러한 미반응된 실란올의 영향은 치환기의 hydrogen bonding acceptor basicity(${\beta}$)가 클수록 크게 나타났고 Phenyl 컬럼보다 $ C_{18}$ 컬럼에서 이들의 영향이 크게 나타났다. 시료의 van der Waals volume(VWV)과 ${\beta}$값을 함께 고려한 파라미터 (1.01VWV/100-1.84${\beta}$)와 시료의 머무름(log k')과는 좋은 직선관계가 있다. 따라서 $ C_{18}$ 및 Phenyl 컬럼에서 페놀 일치환체들의 머무름은 시료의 (1.01VWV/100-1.84${\beta}$)값으로 쉽게 예측할 수 있음을 알았다.

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상압가염형 폴리에스테르 섬유의 물성과 염색성 (The physical properties and the dyeability of the easily dyeable polyester yarn under atmospheric pressure)

  • 김태경;윤석한;신상엽;임용진;조규민
    • 한국염색가공학회지
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    • 제13권6호
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    • pp.33-33
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    • 2001
  • The physical properties and the dyeability of the easily dyeable polyester yarn(EDY) were investigated and compared with those of regular polyester (REG-PET). The EDY, copolymerized with small amount of polyethylene glycol(PEG), showed higher intensity of aliphatic CH peak in IR spectrum, lower density and lower compactness than those of the REG-PET from the analysis of IR, density gradient column and XRD respectively. In the physical properties, the EDY has lowers $T_g,\;T_m$, specific stress and initial modulus, and also has higher strain than that of the REG-PET. The EDY can be dyed under atmospheric pressure and its dyeing rate was faster than REG-PET due to low $T_d$, and this seems to be caused by the increased flexibility of Polymer chain in amorphous region of the EDY due to the copolymerization of PEG.ns being within the experimental error, the average values of lifetim. $\tau$(t) are taken for further calculations. Rate constants such as Stern-Volmer quenching constants K$_{sv}$, quenching rate parameters k$_q$ and k''$_q$, static quenching constant V and kinetic distance r are determined using the modified Stern-Volmer eq.tion and sphere of action static quenching model. In order to see whether the reactions are diffusion limited, equations k$_q$ = e$^{-Eq/RT}$ and k''$_q$ = e$^{-Eq/RT}$ are used to determine the values of E$_q$ and E''$_q$, the activati. energies for collisional quenching and the values of E$_q$ are 14.53, 17.28 and 16.20 kJ mole$^{-1}$ for MPNO1, MPNO2 and 2-PI respectively and the values of E''$_q$ are 14.62 and 17.73 for MPNO1 and MPNO2 respectively. From the magnitudes of various quantities it has.een concluded that the reactions are diffusion limited and the observed positive deviations in the S-V plot are due to static and dynamic quenching.