• Title/Summary/Keyword: cation exchange chromatography

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Operating Parameters for Glutamic Acid Crystallization in Displacement Ion Exchange Chromatography

  • Lee, Kisay
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.2 no.2
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    • pp.117-121
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    • 1997
  • Glutamic acid can be crystallized inside cation exchange column when displacer NaOH concentration is high enough to concentrate displaced glutamic acid beyond its solubility limit. Resulting crystal layer of glutamic acid was moved with liquid phase through the column, and thus could be eluted from the column and recovered in fraction collector. For the purpose of enhancing crystal recovery, effects of operating parameters on the crystal formation were investigated. The increase in the degree of crosslinking of resin favored crystal recovery because of its low degree of swelling. Higher concentration of displacer NaOH was advantageous. If NaOH concentration is too high, however, crystal recovery was lowered due to the solubility-enhancing effects of high pH and ionic strength. The decrease of mobile phase flow rate enhanced crystal recovery because enough time to attain local equilibrium could be provided, but film diffusion would control the overall crystal formation with extremely low flow rate. Lower temperature reduced solubility of glutamic acid and thus favored crystal formation unless the rate of ion exchange was severely reduced. The ion exchange operated by displacement mode coupled with crystallization was advantageous in reducing the burden of further purification steps and in preventing purity-loss resulted from overlapping between adjacent bands.

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Synthesis of $\textrm{N}_{G}$-Mon $o^{14}\textrm{C}$-methyl]-L-arginine ($\textrm{N}_{G}$-Mon $o^{14}\textrm{C}$-methyl]-L-arginine의 합성)

  • 조영봉
    • Environmental Analysis Health and Toxicology
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    • v.1 no.1
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    • pp.77-80
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    • 1986
  • Radiochemical synthesis of $N^{G}$-mono[$^{14}$ C-methyl]-L-arginine is described. The compound was synthesized from radio-active mono[$^{14}$ C]-methylamine as easily and purified by strong cation-exchange resin (NH form) liquid chromatography using a gradient of ammonium hydroxide, and crystallized as flavianate. The free amino acid was successfully prepared by strirring its flavianate and strong anion-exchange resin (OH- form), which could remove the flavianic acid from its salt in water below room temperature. Purity of the compound was tested by thin-layer chromatography, thin-layer electro-phoresis, and scintillation spectrometry.y.

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Study on the of the Correlation between Soil Chemical Properties and Bioactive Compounds of Acer tegmentosum Maxim.

  • Lee, Dong Hwan;Park, Youngki;Hong, Seong Su;Park, Gwang Hun;Kim, Hyun-Jun
    • Korean Journal of Plant Resources
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    • v.34 no.6
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    • pp.566-574
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    • 2021
  • This research was carried out to investigate the correlation between soil chemical properties and bioactive compounds of Acer tegmentosum Maxim. The methods of determining bioactive compounds were determined by high performance liquid chromatography, that contained (-)-gallocatechin (0.04±0.01 ~ 0.43±0.28%), salidroside (0.90±0.06 ~ 3.86±0.59%), tyrosol (0.03±0.00 ~ 0.43±0.00%), (-)-catechin (0.05±0.01 ~ 0.37±0.14%), 6'-O-galloylsalidroside (0.02± 0.01 ~ 0.31±0.06%), (-)-epicatechin-gallate (0.01±0.00 ~ 0.04±0.01%). The soil chemical properties analysis such as soil pH, electric conductivity (EC), organic matter (OM), total nitrogen (TN), available phosphate (Avail. P2O5), exchangeable cation and cation exchange capacity (CEC) were performed following the standard manual. The correlation analysis between soil chemical properties and bioactive compounds of A. tegmentosum, soil pH, available phosphate and exchangeable cation (Ca2+ and Mg2+) were negatively correlated with content of salidroside. On the other hand, soil exchangeable cation (Na+) showed positive correlation with content of salidroside. The results of this study was able to investigate the correlation between soil chemical properties and bioactive compounds of A. tegmentosum.

Formation of methylamine from NG-Monomethyl-L-arginine in Rat (흰쥐에서 NG-Monomethyl-L-arginine으로부터 methylamine의 생성)

  • Cho, Young Bong;Ahn, Young Kon;Choi, Hong Soon;Kim, Choon Sung
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.6 no.1
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    • pp.138-143
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    • 1996
  • After oral administration of 14C-labelled $N^G$-mono[methyl-14C]-L-arginine into rats, 38.2 % and 14.7 % of the administered radioactivity bad been recovered in the urine and stool during 10 days. In the urine, 59.4 % of the radioactivity was recovered in the first 24-hours and used for the indentification of the formation of methylamine. The strong cation-exchange resin column chromatography showed 6.3 %, 7.4 %, 4.9 %, and 81.5 % of the distributions of radioactivity of the neutral, monomethylamine, basic, and uneluted portions, respectively. The radioactivity of monomethylamine portion reeluted into the column chromatography was 39.5 %. The radioactivities corresponding monomethylamine in the column chromatography, thin-layer chromatography, and thin-layer electrophoresis were 39.5 %, 37.3 %, and 28.8 % of the recovered radioactivity, respectively.

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Analysis of Anthocyanin Pigments from Purple-Fleshed Sweet Potato (Jami) (자색고구마(자미) Anthocyanin 색소의 성분 분석)

  • 이란숙;김선재;임종환
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.4
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    • pp.555-560
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    • 2000
  • Anthocyanin pigments of purple-fleshed sweet potato (Ipomoea batatas) were extracted with methanol containing 1% HCL and purified with Amberlite IRC-50 cation exchange resin column chromatography. ndividual pigments were isolated by paper chromatography. Among the four bands obtained by paper chromatography, three major bands were identified to be pure pigments by HPLC system. Two pigments were identified through the analysis of acyl moiety, sugar moiety, alkaline degradation products of aglycone, Rf value of paper chromatogram and retention time of HPLC. The anthocyanin pigments of purple-fleshed weet potato seemed to be composed of peonidin-3-diglucoside-5-glucoside acylated with caffeic or ferulic acids.

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A Convenient Manufacturing Method for Mass Production of EGCG Rich Green Tea Extract (Epigallocatechin Gallate 고함유 녹차추출물의 제조공정 개선)

  • Seo, Eun Hye;Kim, Eun Jeong;Cheon, Seong Bong;Yoon, Min Ji;Choi, Sang Un;Ryu, Geon-Seek;Ryu, Shi Yong
    • Korean Journal of Pharmacognosy
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    • v.50 no.3
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    • pp.198-204
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    • 2019
  • A facile and convenient method was developed for the mass production of epigallocatechin gallate (EGCG) rich green tea extract (Er-GTE). The Er-GTE was successfully obtained from the crude water extract of green tea by the combination of two step purification, i.e., a simple adsorption process on the cation exchange resins (Trilite SCR-B) followed by the chromatography with Diaion HP-20 resins. The green tea extract produced by water extraction under $45^{\circ}C$ was subjected to adsorb on the strongly acidic cation exchange resin, Trilite SCR-B. The eluate passed through the resin was reabsorbed on Diaion HP-20 resin, which was subjected to elute with a mixture of water and alcohol by conventional chromatographical manner. The EGCG content in Er-GTE was estimated above 97% by HP-LC analysis and the newly developed method was regarded as the most suitable and appropriate process for the mass production of epigallocatechin gallate rich green tea extract (Er-GTE).

Moment Analysis (MA) of Lysozyme in Cation Exchange High Performance Liquid Chromatography (HPLC) (양이온교환 고성능액체크로마토그래피에서 라이소자임의 모멘트 분석)

  • Ko, Kwan Young;Kim, In Ho
    • Korean Chemical Engineering Research
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    • v.54 no.4
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    • pp.487-493
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    • 2016
  • The moment analysis of lysozyme was implemented using chromatograms that were obtained from weak cation exchange column in high performance liquid chromatography system. Three elution sodium phosphate buffers containing 1.0, 0.75, 0.5M sodium chloride were used. Experiments were conducted by varying flow rate, elution sodium chloride concentration, and lysozyme solute concentration. The general rate (GR) model was employed to calculate the first moment and the second moment. By plotting $L/u_0$ vs. $({\mu}_1-t_0)/(1-{\varepsilon}_e)(1-{\varepsilon}_i)$] equilibrium constants (K) were obtained from first moment analysis. Intra-particle diffusivity was obtained from theoretical plate number data. Based on the results of moment analysis, van Deemter plots were drawn in order to investigate the contributions of $H_{ax}$, $H_f$, and $H_d$ to total Height Equivalent to a Theoretical Plate (HETP, $H_{total}$). The effect of intra-particle diffusion ($H_d$) was the most dominant factor contributing to HETP while external mass transfer ($H_f$) was negligible factor.

Studies on Ion-exchange Chromatography of Elements in Special Nonferrous Alloys (비철특수합금에서 금속원소의 이온교환 크로마토그라프에 관한 연구)

  • Kyung Woong Lee;Young Jin Yoo
    • Journal of the Korean Chemical Society
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    • v.29 no.2
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    • pp.178-182
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    • 1985
  • The purpose of this study was to develop a separation method of Zn(II), Cu(II) and Mg(II), by ion exchange chromatography using cation exchange resion (Dowex 50w${\times}$8, 80-100 mesh) and anion exchange (Amberlite IRA-400). Ion exchange resions were packed into 25 ${\times}$ 2cm ID column and flow rate was controlled to 0.30 ml/min. Good eluents for separation of nonferrous metal ions such as Zn(II), Cu(II), Mg(II) were as follow: 0.5M $NaNO_3$ (pH 3.1), 0.2~0.5M HCl + 50~60% Acetone, and 1M HAc + 0.1M NaAcf(pH 3.7) aqueous solution. The mixed solution of 0.1M NaAc(pH 3.7), 0.5M HCl + 50% Acetone were found to be the best eluent for step elution. Analysis of metals were determined by atomic absorption spectrophotometer. In addition, separated Zn(II) fraction was obtained by eluted with 0.12N HCl and 1.5N $NH_4OH$ aqueous solution. This solution was titrated by the E. D. T. A.

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Purification of Cellulase from Trichoderma viride and properties of Its Component Enzymes

  • Dong Won Kim;Tae Seung Kim
    • Bulletin of the Korean Chemical Society
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    • v.15 no.9
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    • pp.719-724
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    • 1994
  • Major cellulase components, such as three endoglucanases (endoglucanases I, II, and III) and one exoglucanase (exoglucanase II), were isolated from a commercial cellulase (Meicelase TP 60) derived from the fungus Trichoderma viride by a series of chromatography procedures. These procedures were the gel filtration on Bio-Gel, the anion exchange on DEAE-Bio-Gel A, the cation exchange on SP-Sephadex C50, and the affinity chromatography on Avicel cellulose. The average molecular weights determined by SDS-polyacrylamide gel electrophoretic analysis were 51,000, 59,000, 41,000 and 62,000 Da for endoglucanases I, II and III and exoglucanase II, respectively. The extinction coefficients, ${\varepsilon}^{1%}$ 280 nm, of these enzymes were 11.7, 3.3, 7.2 and 11.3, respectively. Among them, the endoglucanase II showed the very low value of the coefficient compared with the others. On the other hand, it was found that endoglucanase II and III were of more random hydrolytic mode on carboxymethylcellulose as compared with those of endoglucanase I and exoglucanase II. Especially, endoglucanase I showed less random action than that of exoglucanase II. In the hydrolysis of insoluble cellulose by the enzyme components, cellobiose was the major product, but glucose was the major product by endoglucanase III.

Purification and Characterization of Alcohol Dehydrogenase from Acetobacter sp. KM (Acetobater sp.KM Alcohol Dehydrogenase의 분리 및 특성)

  • 전홍성;차영주
    • KSBB Journal
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    • v.10 no.1
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    • pp.30-37
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    • 1995
  • Membrane-bound alcohol dehydrogenase(ADH) was purified to homogeneity from the acetic acid producing bacteria, Acetobacter sp. KM. The enzyme was solubilized and extracted with Triton X-100 and purified using the Mono-Q ion exchange chromatography and Superose 12 gel filtration chromatography. The enzyme was purified to 12-fold with a yield of 30%. The molecular weight of the purified enzyme was to be 335 KDa. SDS-PAGE of the enzyme showed two subunits with molecular weights of 79 KDa and 49 KDa. It indicated that the enzyme consisted of three subunits of the 79 KDa and two subunits of the 49 KDa. The purified .ADH preferentially oxidized straight chain aliphatic alcohol except methanol. Formaldehyde, acetaldehyde and glutaraldehyde were also oxidized. The apparent Km for ethanol was 1.04 mM and the optimum pH and temperature were 5.0∼6.0 and 32$^{\circ}C$, respectively. V2O5 and divalent cation such as ZnCl2 and NiCl2 inhibited enzymatic activity.

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