• Title/Summary/Keyword: ion exchange chromatography

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Simultaneous Determination of Anions and Cations in Natural Water by Ion-exclusion/Cation-exchange Chromatography with a Weakly Acidic Cation-exchange Resin Column

  • Lee, Kwang-Pill;Choi, Seong-Ho;Park, Yu-Chul;Bae, Zun-Ung;Lee, Mu-Sang;Lee, Sang-Hak;Chang, Hye-Yong;Kwon, Se-Mok;Kazuhiko Tanaka
    • Bulletin of the Korean Chemical Society
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    • v.24 no.9
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    • pp.1324-1328
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    • 2003
  • The simultaneous determination of anions ($SO_4 ^{2-},\;Cl^-,\;and\;NO_3^-$) and cations ($Na^+,\;NH^{4+},\;K^+,\;Mg^{2+},\;and\;Ca^{2+}$) in natural water obtained by Nakdong River waters system in Korea were performed by ion-exclusion/cationexchange chromatography with conductimetric detection. The stationary phase was a polymethacrylate-based weakly acidic cation-exchange resin column in the $H^+$-form and a weak-acid eluent. When using only a 1.4 mM sulfosalicylic acid/6 mM 18-crown-6 ether as an eluent, good resolution of both anions and cations, minimum time required for the separation, and satisfactory detection sensitivity were obtained in a reasonable time. The method was successfully applied to the simultaneous determination of anions and cations in natural waters.

Subunits of Sweet Potato $\beta$-Amylase (고구마 $\beta$-아밀라제의 subunit)

  • 안용근
    • The Korean Journal of Food And Nutrition
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    • v.1 no.2
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    • pp.50-55
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    • 1988
  • B-Amylase was obtained from sweet potato extract in a crystalline state by dialysis against water after precipitated with acetone according to the method reported previously followed by DEAE Sephadex A-50 ion exchange chromatography plus gel chromatography of Sephadex G-200. The purified enzyme was homogeneous by SDS PAGE. The efforts had done to remove the miner bands in SDS PAGE by Sephadex G-200 gel chromatography, DATE Sephadex A-50 ion exchange chromatography. isoelectrophocusing, affinity chromatography, hydroxyapatite chromatography, recrystallizstion and HPLC on a column of TSK gel SW 3000 but have given any result. But, N -terminal amino acid of the enzyme was revealed mainly alanine and trace of glycine and glutamic acid. Therefore, it seems that the miner bands in SDS PAGE have a role of subunit.

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Purification and Partial Amino Acid Sequence of a Bacteriocin Produced by Lactococcus, sp. HY449 (Lactococcus sp. HY449가 생산한 Bacterisocin의 정제)

  • 오세종;이상준;김경태;김상교;박연희;백영진
    • Microbiology and Biotechnology Letters
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    • v.29 no.3
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    • pp.155-161
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    • 2001
  • A bactriocin produced by Lactrococcus sp. HY449 was purified by sequential purfication steps such as n-propanol-acetone precipitation ion -exchange chromatography using CM-Sequential CL6B. gel filtration chromatography using Sephacry HR100 and reverse-phase chromatography using pro RPC HR 5/10. Reverse-phase chromatography the final step of the purfication yielded a single symmetrical peak of bacteriocin activity The purification resulted in final yield of 3.25% and 413.35 fold increase of the specific activity of bacteriocin. The active fraction from reverse-phase chromatography was used for N-terminal amino acid analysis . The purified bacteriocin contained isoleucine, leucine, methionine, and glycine at but N-terminal end no aromatic amino acids. Calculation of the number of amino acid residues in the bacteriocin revealed that it is consisted of 32 residues assuming the molecular weight of bacteriocin to be about 3.6kDa. Edman degrandation elucidated amino acid residues of the first four of the N-terminus to be $NH_2$-Ile-Leu-Pro-GIn.

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Purification and Characterization of ATPase and Phosphatase of Light Membrane Vesicles Isolated from Cucurbita pepo (Cucurbita pepo에서 분리한 Light Membrane Vesicle의 ATPase와 Phosphatase의 정제 및 특성)

  • 오승은
    • Journal of Plant Biology
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    • v.33 no.4
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    • pp.325-332
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    • 1990
  • Light membrane vesicles were isolated from the zucchini hypocotyl by floatation on ficoll density gradients and the proteins were solubilized with Triton X100. Three ATP-hydrolyzing enzymes were partially purified by ion-exchange and gel filtration chromatography and isoelectric focusing. There are plasma membrane-type ATPase whose activity was inhibited by vanadate but not by nitrate, tonoplast-type ATPase which was sensitive to nitrate but insensitive to vanadate and one having a phosphatase activity with a pI value different from that of an acid phosphatase. A fraction was obtained after DEAE-ion-exchange chromatography crossreacting with polyclonal antibodies against Ca2+ -ATPase from human erythrocytes.

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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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Purification and Characterization of the Regulatory Substance of Furfural Biodegradation in Pseudomonas fluorescens (Pseudomonas fluorescens에 의한 Furfural의 분해대사 조절물질에 관하여)

  • 이병웅;유병설;이계준;하영칠
    • Korean Journal of Microbiology
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    • v.23 no.4
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    • pp.241-247
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    • 1985
  • The objectives of this study were to isolate and identify ninhydrin positive substande(s) produced in the culture broth of Pseudomonas fluorescens. It was found that the NPS could stimulate bioconversion of furfural into furoic acid. In order to isolate the NPS from the culture broth, cell free filtrate was subjected to ion-exchange chromatography, gel-permeation and finally to cellulose column chromatography. The purified NPS was white amorphous power and very soluble in water, slightly soluble in methanol and very insoluble in organic solvents. UV, and IR absorption spectra. $^H$ and $^{13}C-NMR$ were measured in order to identify the chemical structure of the NPS.

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A Study of Protein Ion Exchange Chromatography based on Plate Theory (단이론에 따른 단백질 이온교환 크로마토그라피의 연구)

  • 김인호;김진태
    • Microbiology and Biotechnology Letters
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    • v.23 no.4
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    • pp.485-491
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    • 1995
  • Protein ion exchange chromatography was studied experimentally in order to prove the theoretical prediction from the linear model of Yamamoto, S. et al (1). Adsorption isotherms were measured as a function of ionic strength in a batch experiment. The relationship between the characteristics of chromatogram and the operating conditions of ionic strength, flow rate, length of column, concentration and amount of protein sample were studied. At the higher ionic strength, the lower flow rate and the longer column conditions, the higher number of plate was obtained. Satisfactory agreement was observed between the experimental and the calculated chromatograms except for the case of high protein concentration.

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Separation Characteristics of IgY (Immunoglobulin Yolk) in Various HPLC Columns (다양한 HPLC Column에서의 IgY(Immunoglobulin Yolk) 분리특성)

  • Song, Sung Moon;Kim, In Ho
    • Korean Chemical Engineering Research
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    • v.50 no.4
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    • pp.659-665
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    • 2012
  • IgY (Immunoglobulin Yolk) in egg yolk corresponds to IgG (Immunoglobulin G) in animal serum and plays an important role as immunological proteins in intestines. Carrageenan and Arabic gum were used as pretreatment agents to purify IgY from fresh egg yolk. DEAE (Diethylaminoethyl) Sepharose column in FPLC (Fast Protein Liquid chromatography) was an ion exchange tool to remove contaminants as well as to elute IgY from the column. GF HPLC (Gel Filtration High Performance Liquid Chromatography) enables to measure the molecular weights of IgY and to identify the purified IgY by comparing the molecular weight of standard IgY with the purified one. IgY is a heterogeneous group of different molecular weight and ionic properties, which was investigated with various IE HPLC (Ion Exchange High Performance Liquid Chromatography) columns such as AX, CX and SCX. Three peaks of IgY were separated in the AX column under the conditions of 0.5 M NaCl and pH=8. The SCX column also gave the three peaks of IgY at 0.5 M NaCl and pH=5.

Chromatographic Enrichment of Lithium Isotopes by Hydrous Manganese(IV) Oxide

  • Kim, Dong Won
    • Bulletin of the Korean Chemical Society
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    • v.22 no.5
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    • pp.503-506
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    • 2001
  • Separation of lithium isotopes was investigated by chemical ion exchange with a hydrous manganese(IV) oxide ion exchanger using an elution chromatography. The capacity of manganese(IV) oxide ion exchanger was 0.5 meq/g. One molar CH3COO Na solution was used as an eluent. The heavier isotope of lithium was enriched in the solution phase, while the lighter isotope was enriched in the ion exchanger phase. The separation factor was calculated according to the method of Glueckauf from the elution curve and isotopic assays. The single stage separation factor of lithium isotope pair fractionation was 1.021.