• Title/Summary/Keyword: anion exchange column

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A Multidimensional System for Phosphopeptide Analysis Using TiO2 Enrichment and Ion-exchange Chromatography with Mass Spectrometry

  • Cho, Kun;Yoo, Ji-Sun;Kim, Eun-Min;Kim, Jin-Young;Kim, Young-Hwan;Oh, Han-Bin;Yoo, Jong-Shin
    • Bulletin of the Korean Chemical Society
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    • v.33 no.10
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    • pp.3298-3302
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    • 2012
  • Although offline enrichment of phosphorylated peptides is widely used, enrichment for phosphopeptides using $TiO_2$ is often performed manually, which is labor-intensive and can lead to irreproducible results. To address the problems associated with offline enrichment and to improve the effectiveness of phosphopeptide detection, we developed an automated online enrichment system for phosphopeptide analysis. A standard protein mixture comprising BSA, fetuin, crystalline, ${\alpha}$-casein and ${\beta}$-casein, and ovalbumin was assessed using our new system. Our multidimensional system has four main parts: a sample pump, a 20-mm $TiO_2$-based column, a weak anion-exchange, and a strong cation-exchange (2:1 WAX:SCX) separation column with LC/MS. Phosphorylated peptides were successfully detected using the $TiO_2$-based online system with little interference from nonphosphorylated peptides. Our results confirmed that our online enrichment system is a simple and efficient method for detecting phosphorylated peptides.

Isolation and Characterization of Exogenously Expressed Calmodulin from Endogenous Tobacco Calmodulin by Anion-exchange Fast Protein Liquid Chromatography

  • Oh, Suk-Heung;Cha, Youn-Soo;Lee, Tae-Kyoo
    • BMB Reports
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    • v.28 no.4
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    • pp.306-310
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    • 1995
  • A Mono Q HR 5/5 anion-exchange column with a FPLC system was used to separate exogenously expressed calmodulin from endogenous tobacco calmodulins. Transgenic tobacco calmodulins were purified by fractionation with ammonium sulfate, precipitation with sulfuric acid and hydrophobic chromatography on phenyl-Sepharose CL-4B. The purified calmodulins were chromatographed in the FPLC using the column. This method was selected because of the slight differences in the net charge of foreign and endogenous plant calmodulins due to amino acid sequence differences. By this approach, the exogenously expressed calmodulin was isolated from endogenous tobacco calmodulins. The isolated calmodulin was characterized by amino acid composition analysis as well as methylation analysis.

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Determination of Impurities in Uranium Dioxide by Neutron Activation Analysis (중성자방사화분석법에 의한 이산화우라늄중의 불순물정량)

  • Nak Bae Kim;Hae-Ill Bak;Chul Lee
    • Nuclear Engineering and Technology
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    • v.13 no.4
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    • pp.237-244
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    • 1981
  • The preliminary concentration of trace elements in uranium dioxide using an anion exchange resin is presented for neutron activation analysis. The uranyl solution in sulfuric acid is adjusted to the acidity of about pH 2.7 and loaded on a column of the anion exchange resin. An appropriate volume of eluates obtained from the column shows good recoveries of trace elements. By combining this preconcentration with a radiochemical separation scheme, which was developed for the determinations of impurities in aluminum, it is possible to determine 21 trace elements in reactor grade uranium dioxide.

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Separation of Goid, Palladium and Platinum in Chromite by Anion Exchange Chromatography for Inductively Coupled Plasma Atomic Emission Spectrometric Analysis

  • Choe, Gwang Sun;Lee, Chang Hyeon;Park, Yeong Jae;Jo, Gi Su;Kim, Won Ho
    • Bulletin of the Korean Chemical Society
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    • v.22 no.8
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    • pp.801-806
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    • 2001
  • A study has been carried out on the separation of gold, iridium, palladium, rhodium, ruthenium and platinum in chromite samples and their quantitative determination using inductively coupled plasma atomic emission spectrometry (ICP-AES). The dissolution condition of the minerals by fusion with sodium peroxide was optimized and chromatographic elution behaviour of the rare metals was investigated by anion exchange chromatography. Spectral interference of chromium, a matrix of the minerals, was investigated on determination of gold. Chromium interfered on determination of gold at the concentration of 500 mg/L and higher. Gold plus trace amounts of iridium, palladium, rhodium and ruthenium, which must be preconcentrated before ICP-AES was separated by anion exchange chromatography after reducing Cr(Ⅵ) to Cr(III) by H2O2. AuCl4- retained on the resin column was selectively eluted with acetone- HNO3-H2O as an eluent. In addition, iridium, palladium, rhodium and ruthenium remaining on the resin column were eluted as a group with concentrated HCl. However, platinum was eluted with concentrated HNO3. The recovery yield of gold with acetone-HNO3-H2O was 100.7 ${\pm}2.0%$, and the yields of palladium and platinum with concentrated HCl and HNO3 were 96.1 ${\pm}1.8%$ and 96.6 ${\pm}1.3%$, respectively. The contents of gold and platinum in a Mongolian chromite sample were 32.6 ${\pm}$ 2.2 ${\mu}g$/g and 1.6 $\pm$ 0.14 ${\mu}g$/g, respectively. Palladium was not detected.

A Study on Ion Exchange Characteristics with Composition and Concentration of Electrolyte, Ratio of Ion Exchange Resin (전해질 성분 및 농도, 이온교환 수지 비율에 따른 이온교환 특성 연구)

  • Ahn Hyun-Kyoung;Rhee In-Hyoung;Yoon Hyoung-Jun;Jeong Hyun-Jun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.7 no.4
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    • pp.727-732
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    • 2006
  • The object of this study was to investigate the influence of composition and concentration of electrolyte, ratio of cation to anion exchange resin of mixed ion exchange column in the performance of ion exchange. Also this work examined the removal capability of suspended solids by ion exchange resin and the effect of particule on the characteristics of ion exchange. Breakthrough time was extended as the amount of ions and particles present in liquid was decreased. The case of anion, the breakthrough sequence is $Cl^{-}, but the case of cation, the breakthrough sequence is $Na^{+}. As for the ratio of cation to anion exchange resin of 1:2, the breakthrough time was prolonged compared with that of 1:1 and 1:3. For the electrolyte of equal concentration containing suspended solid, breakthrough time was contracted less than 20%. It results in the increase in the removal capacity of cation exchange resin. For the higher ratio of cation exchange resin, suspended solids are shorten the cation's breakthrough time so that the runtime of ion exchange resin tower is increased.

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The Mixed-Bed Ion Exchange Performance and Temperature Effects at Ultra-Low Concentrations - 2.Temperature Effects - (초저이온 농도범위에서 혼합층 이온교환능과 온도의 영향 - 2. 온도의 영향 -)

  • Yoon, Tae Kyung;Noh, Byeong Il;Lee, Chang Won;Moon, Byung Hyun;Lee, Gang Choon;Jo, Myung Chan
    • Applied Chemistry for Engineering
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    • v.10 no.2
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    • pp.206-211
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    • 1999
  • Mixed-bed ion exchange performance was studied experimentally with variations of cation to anion resin ratio, resin weight and temperature at ultralow sodium chloride solution concentrations of less than $1.0{\times}10^{-4}M$. Analyzing the effluent concentration histories the performance test was examined as a function of tested solution volume for a laboratory-scale continuous flow column until both the cation and anion-exchange resins were exhausted. Initial leakage was observed for both cation and anion breakthrough curves, but serious at cation breakthrough curve because of low selectivity coefficient. The slope of breakthrough curve was affected by selectivity coefficient and temperature. The slope of anion breakthrough curve was steep because of the large selectivity coefficient, and ion exchange rates increased as temperature increased. The temperature effect decreased as the total volume was increased or as the resins were exhausted.

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Isolation and Purification of Fibrinolytic Enzyme of Edible Mushroom, Sarcodon aspratus(Berk.)S. Ito (능이버섯으로부터 Fibrin 분해활성이 있는 단백질의 분리 및 정제)

  • 이종호;양정례;정청송;김희숙;조재선
    • Journal of Life Science
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    • v.11 no.6
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    • pp.561-567
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    • 2001
  • To isolate and purify fibrinolytic active substance from Sarcodon aspratus(N $H_4$)$_2$S $O_4$ precipitation, DE52 anion exchange column chromatography, Sephacryl-S 200gel filtration chromatography and Mono S cation FPLC were carried out and the characterizations of the purified enzyme were investigated. The bound active fraction on DE52 anion exchange column chromatography were eluted with 0.2 M NaCI and the fibrionlytic enzyme was purified after following Sephacryl-S200 gel fitration chromatography and Mono S cation EPLC. The specific activity of purified enzyme was 55.2 U/mg protein and increased 11.3 fold comparing crude extract and the yield was 49.5%. 12% SDS-PAGE electrophoresis and gel filtration chromatography revealed that Sarcodon aspratus fibrionloytic enzyme was highly purified and had 29.300 Da molecular weight. Enzyme activity of the purified fibrinolytic enzyme from Sarcodon aspratus was increased on higher pH and was stable until pH 10.5. On temperature dependent stability, the enzyme activity was decrease sharply but remained 25% relative activity on 8$0^{\circ}C$. This enzyme activity was inhibited by heavy metal ion, C $U^{2+}$ and $Co^{3+}$ with 68% and 38%, respectively. And also, the enzyme activity was inhibited with $Ca^{2+}$ chelator EDTA and serine protease inhibitor PMSF. These results from this study suggested that the fibrinolycit enzyme from Sarcodon aspratus is a serine protease and the enzyme activity was increased by $Ca^{2+}$ or $Mg^{2+}$ ion.n.ion.n.

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Removal of Orthophosphate Ions from Aqueous Solutions Using the Anion Exchange Resin in the Form of $Cl^-$ Ion ($Cl^-$ 형태의 음이온 교환 수지를 이용한 오쏘인산 이온의 제거에 관한 연구)

  • Kim, Ki-Chul;Park, Su-Jin;Cha, Ran;Jeong, Tae-Young;Chung, Hyung-Keun
    • Journal of Korean Society of Environmental Engineers
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    • v.34 no.3
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    • pp.162-167
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    • 2012
  • The removal of orthophosphate ions from aqueous solutions by the anion exchange resin in the form of $Cl^-$ ion was investigated to elucidate the ion exchange mechanism which depends on the forms of orhthophoshate ions. In addition, the effects of alkalinity and other common anions were studied. The results showed that the orhthophosphate ions with the oxidation state of 2 and 3 ($HPO{_4}^{2-}$ and $PO{_4}^{3-}$) were effectively removed by the anion exchange resin, whereas the part of the $H_2PO_4{^-}$ ion passed through the ion exchange column. This suggested that the affinity of $H_2PO_4{^-}$ to the ion exchange resin was comparable with that of $Cl^-$ ion. In all cases, the effluent pHs have shown to be much lower than the calculated values, indicating that more $Cl^-$ ions than the orthophosphate equivalents in the influent were eluded. As the alkalinity increases, the decrease in pH was minimized. When the alkalinity was 100 mg/L ($CaCO_3$) or greater, 100 mg/L orthophosphate ions including $H_2PO_4{^-}$ were completely removed. The common anions such as $SO{_4}^{2-}$ and $NO_3{^-}$ were also removed by the anion exchange resin, and thus decreased the ion exchange capacity for the removal of orthophosphate.

Purification of the Vacuolar Arginine Transporter from Neurospora crassa (Neurospora crassa로부터 arginine transporter의 순수분리)

  • ;Weiss, R. L.
    • Korean Journal of Microbiology
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    • v.27 no.2
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    • pp.117-123
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    • 1989
  • Radioactive N-$\alpha$-p-nitrobenzoxycarbonyl (NBZ)-L-[2,$3-^{3}$H] arginyl diazomethane was used as an affinity label for the vacuolar arginine transporter in Neurospora crassa. Vacuolar matrix proteins were removed by fracturing the membranes with freeze-thaw method in dry ice/ethanol bath. Vacuolar membrane proteins were then wasged with 500mM NaCl to remove ionically bound derivatives and peripheral membrane proteins from vacuolar membranes. After dissolved in 1% Titon X-100, dissolved vacuolar memvrane proteins were separated with molecular sieve column chromatography, anion and cation exchange chromatographies. The arginine transporter was purified giving the purification factor of 1136.

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Determination of Glyphosate in Whole Blood by HPLC-fluorescence Detection (HPLC 형광검출법에 의한 Glyphosate의 혈중농도 측정)

  • 이상기;김기욱;양자열;인상환;이수연
    • YAKHAK HOEJI
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    • v.45 no.4
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    • pp.347-351
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    • 2001
  • A rapid and sensitive method for the determination of glyphosate, a phosphated amino acid herbicide, in whole blood is presented. After removal of protein, the whale blood was purified by using the anion exchange resin (Dowex 1), and derivatized with 9-fluorenylmethyl chloroformate (FMCL). Derivatized glyphosate from blood sample was injected onto a Whatman partisil 10SAX column and separated with 0.1M phosphate buffer (pH 2.5) and acetonitrile (ratio=3:1). The high performance liquid chromatography-fluorescence detection gave the detection limit of 86pg and linearity of 0.9999 in the range of 0.25 $\mu$g/ml and 25 $\mu$g/ml. The recoveries of glyphosate added to the blood samples were ranged from 75.3% to 100.4% compared to the samples prepared in water. The derivatized glyphosate was stable at various acidity and temperature. This method has been successfully applied to the blood samples of lethal intoxication with the herbicide glyphosate.

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