• Title/Summary/Keyword: 이온크로마토그래피

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Developement of Automatic Microanalysis System of Ammonia Gas in Clean Room by Using of Wet Denuder (디누더를 이용한 클린룸 내 미량 암모니아 가스 자동 측정 시스템 개발)

  • 이응선;김정수;김광영;유승교
    • Proceedings of the Korea Air Pollution Research Association Conference
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    • 2000.11a
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    • pp.101-102
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    • 2000
  • 반도체 생산에서 암모니아는 정밀성을 요구하는 공정 특히, 광증폭 공정에서 산과 반응하여 불량을 일으키는 주 원인이며, 고메모리로 갈수록 청정도 기준이 엄격해져서 1 ppbv 이하의 농도를 검출할 수 있는 기술이 요구되고 있다. 현재 반도체 회사에서의 암모니아 분석은 임핀저를 이용하여 가스를 포집한 후 이온크로마토그래피로 분석 하고 있으나, 이는 많은 시간과 인력이 요구되며, 실시간 분석이 어렵기 때문에 반도체 클린룸의 관리에 어려움이 있다. (중략)

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Identification of Proteins in Egg White Using Ion Exchange Cartridge and RP-HPLC (이온교환 카트리지와 RP-HPLC를 이용한 난백 단백질의 확인)

  • Kim, Hyun Moon;Kim, Ah Reum;Lee, Chang Soo;Kim, In Ho
    • Korean Chemical Engineering Research
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    • v.50 no.4
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    • pp.713-717
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    • 2012
  • Approximately forty proteins in egg white have been widely studied for their functional properties. To develop a procedure of separation for pure and non-altered proteins from egg white, purification study was conducted to isolate lysozyme, ovotransferrin, and ovalbumin. Ion exchange cartridge can selectively separate proteins from egg white, and reversed-phase HPLC (RP-HPLC) could identify separated proteins. Proteins in egg white were purified by HI trap ion exchange cartridge SP and Q with buffers pH 8.0 and 5.2. C18 column (Phenomenex, USA) was used for RP-HPLC analysis and isocratic mobile phase was used with acetonitrile (ACN)/distilled water (DW)/trifluoroacetic acid (TFA) in the ratio of 50/50/0.1. Comparing the retention times of standards in RP-HPLC experiments showed that ovotransferrin, ovalbumin, and lysozyme in egg white were eluted successively in the RP-HPLC column after the pretreatment in SP and Q ion exchange cartridges.

Quantitative Real-Time PCR of Porcine Parvovirus as a Model Virus for Cleaning Validation of Chromatography during Manufacture of Plasma Derivatives (혈장분획제제 제조공정에서 크로마토그래피 세척 검증을 위한 모델바이러스로서의 Porcine Parvovirus 정량)

  • Kil Tae Gun;Kim Won Jung;Lee Dong Hyuk;Kang Yong;Sung Hark Mo;Yoo Si Hyung;Park Sue-Nie;Kim In Seop
    • Korean Journal of Microbiology
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    • v.41 no.3
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    • pp.216-224
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    • 2005
  • Chromatography has now been used successfully to provide the requisite purity for human plasma-derived biop-harmaceuticals such as coagulation factors and immunoglobulins. Recently, increasing attention has been focused on establishing efficient cleaning procedures to prevent potential contamination by microorganisms as well as carry-over contamination from batch to batch. The purpose of present study was to develop a cleaning validation system for the assurance of virus removal and/or inactivation during chromatography process. In order to establish an assay system for the validation of virus clearance during chromatography cleaning process, a quantitative real-time PCR method for porcine parvovirus(PPV) was developed, since PPV, a model virus for human parvovirus B19, has a high resistance to a range of physico-chemical treatment. Specific primers for amplification of PPV DNA was selected, and PPV DNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be 1.5 $TCID_{50}/ml$. The established real-time PCR assay was successfully applied to the validation of PPV removal and cleaning during SP-Sepharose cation chromatography for thrombin purification and Q-Sepharose anion chromatography for factor VIII purification. The comparative results obtained by real-time PCR assay and infectivity titrations suggested that the real-time PCR assay could be a useful method for chromatography cleaning validation and that it could have an additive effect on the interpretation and evaluation of virus clearance during the virus removal process.

Experimental and Simulation Study of Lysozyme Separation in Cation Exchange Chromatography (양이온교환 크로마토그래피에서 Lysozyme 분리 실험과 전산모사)

  • Kim, Jung-Ae;Seong, Yeon-Kyeong;Kim, In-Ho
    • KSBB Journal
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    • v.21 no.3
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    • pp.220-223
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    • 2006
  • Lysozyme is an important antibacterial material, as effective food preservative. A number of lysozymes are found in nature such as egg white, where exists about 3.5% of egg proteins. In this study, carboxymethyl cation exchange chromatography has been used for separation of lysozyme. A simulation study by ASPEN was also performed for saving time and cost in chromatography purification experiments. Important parameters in experimental chromatography were sample loading amount, NaCl concentration, and pH of eluent. Simulation results were successfully fitted with chromatograms from experiments with change of parameters mentioned above.

Purification and Characterization of Superoxide Dismutase in Sphingomonas sp. KS 301 (Sphingomonas sp. KS 301의 Superoxide Dismutase 정제 및 특성)

  • Kang, Hee-Jeong;Jeong, Jae-Hoon;Choi, Ji-Hye;Son, Seung-Yeol
    • Korean Journal of Microbiology
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    • v.43 no.2
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    • pp.83-90
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    • 2007
  • Sphingomonas sp. KS 301, which was isolated from oil contaminated soil, was shown to have five different SODs (SODI, II, III, IV, V) which can be separated by DEAE-Sepharose chromatography, and SOD III was finally purified in this study by ammonium sulfate precipitation, DEAE-Sepharose chromatography, Superose 12 gel filtration and Uno-Q1 ion exchange chromatography. The molecular weight of SOD III was 23 kDa as determined by SDS-PAGE and the apparent molecular weight of the native enzyme was estimated to be approximately 71 kDa by Superose-12 gel filtration chromatography. These data suggest that the purified SOD consists of at least two subunits. The specific activity of the SOD III was higher than Mn type or Fe type SOD of Escherichia coli by 5 fold. To determine the type of SOD III, inhibitory effects of $NaN_{3},\;H_{2}O_{2},\;KCN$ were examined. 10 mM $NaN_{3}$ was able to inhibit 56% of the SOD III activity, which indicates that this SOD is Mn type. The optimum pH of the SOD III was 7.0 and the optimum temperature was $20^{\circ}C$. N-terminal amino acid sequence of purified SOD III was most similar to those of Psudomonase ovalis and Vibrio cholerae among bacteria.

Determination of Some Inorganic Anions in Saline Water by Ion Chromatography with UV Detection (이온크로마토그래피를 이용한 소금물중의 무기음이온들의 분리정량)

  • Han, Sun Ho;Park, Yang Soon;Park, Soon Dal;Joe, Kih Soo;Eom, Tae Yoon
    • Analytical Science and Technology
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    • v.12 no.2
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    • pp.99-104
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    • 1999
  • A stepwise gradient elution with two wavelengths detection was performed for the separation and determination of some anions in saline water. The eight anions such as iodate, bromate, nitrite, bromide, nitrate, chromate, iodide and thiocyanate were successfully separated using AS-7 column and sodium chloride/sodium phosphate buffer solution as an eluant within 40 min. The separation behaviors of anions were studied at various sodium chloride concentrations. The peak shapes of anions of bromate, nitrite, bromide and nitrate gradually broadened as the concentration of sodium chloride increased until 1.0 M in the sample solutions. However, no effect was observed in the peak shapes of chromate, iodide and thiocynate. A good linearity was obtained at the range of ppm(mg/L). The detection limit was proved to be $10-720{\mu}g/L$ for the eight anions with $50{\mu}L$ injection volume. This method was applied to the determination of $Br^-$, ${NO_3}^-$ and $I^-$ in sea water.

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Determination of Glutathione in Biological Samples by Ion-pairing HPLC/FLD (이온쌍 HPLC/FLD를 이용한 생체 시료중의 Glutathione 농도 분석)

  • Yoo, Jeong-Yeon;Lee, Kyoung-Ok;Shin, Ho-Sang
    • Analytical Science and Technology
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    • v.12 no.1
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    • pp.28-33
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    • 1999
  • Glutathione(GSH) in biological samples was determined by high performance liquid chromatographic(HPLC) method with fluorescence detector(FLD) after monobromobimane(MBB) or 4-fluoro-7-sulfobenzofurazan(SBD-F) derivatization. The detection limit of $0.03{\mu}g/mL$ was obtained after MBB derivatization, derivative of MBB was about 200 times more sensitive than that of SBD-F. N-acetylcysteine was used as internal standard and tetrabutylammonium ion as counter ion for better separation. The determination by ion-pairing chromatography after MBB derivatization was characterized by linearity in the range between $0.08{\sim}8.33{\mu}g/mL$ with a good correlation coefficient of 0.998. By precision test appeared relative standard deviation at less than 5% at three different concentrations. This method can be used for the analysis of GSH in plasma and tissue.

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Induced Circular Dichroism Spectra and Chromatography by Interaction of Some Racemic Cobalt (III) Complexes and d-Tartrate$^2$- in Aqueous Solution (d-주석산이온과 라세미코발트(III) 착물과의 상호작용에 따른 유발원편광이색성 Spectra와 크로마토그래피)

  • Chang Eon Oh;Dae Ho Kang;Gab Choul Shin
    • Journal of the Korean Chemical Society
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    • v.25 no.5
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    • pp.306-310
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    • 1981
  • Induced circular dichroism spectra of racemic cobalt(III) complexes for $[Co(en)_3]^{3+},\;[Co(tn_)3]^{3+},\;cis-[Co(NH_3)(en)_2]^{3+},\;[Co({\beta}-ala)(en)2_]^{2+},\;[Co(gly)(en)_2]^{2+}\;and\;[Co(acac)(en)_2]^{2+}$ were measured when they were dissolved in aqueous d-tartrate2- solution at room temperature. Only a single negative CD spectrum was observed for all the complexes above in visible region(400∼500nm). It was interpreted that these CD bands were attributed to the difference in interaction between ${\Lambda}$-and ${\Delta}$-enantiomers with d-tartrate$^{2-}$. Namely, when d-tartrate$^{2-}$ was added to ${\Lambda}$-enantiomer and ${\Delta}$-enantiomer, it caused ${\Lambda}$-enantiomer to change greatly and ${\Delta}$-enantiomer to change only slightly; combined the results proved induced circular dichroism. The enantiomer for which the eluent has a stronger affinity should be eluted faster in ion-exchange column chromatography. It is possible to predict the elution order of chromatography from the sign of the induced CD if stronger interaction of chiral anion with the complex leads to greater change in the natural CD spectrum of the complex. The elution order was in complete agreement with the prediction from the sign of the induced CD spectrum for all the measured complexes.

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Separation for the Determination of $^{59/63}Ni$ in Radioactive Wastes (방사성 폐기물 내 $^{59/63}Ni$ 정량을 위한 분리)

  • Lee, Chang-Heon;Jung, Kie-Chul;Choi, Kwang-Soon;Jee, Kwang-Young;Kim, Won-Ho
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.3 no.4
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    • pp.309-317
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    • 2005
  • A study on the separation of $^{99}Tc,\;^{94}Nb,\;^{55}Fe,\;^{90}Sr\;and\;^{59/63}Ni$ in various radioactive wastes discharged from nuclear power plants has been performed for a use in their quantification which is indispensible for the evaluation of the radionuclide inventory Ni was recovered along with Ca, Mg, Al, Cr, Ti, Mn, Ce, Na, K, and Cu through the sequential separation procedure of Re(as a surrogate of $^{99}Tc$), Nb, Fe and Sr by anion exchange and Sr-Spec extraction chromatography. In this research, chemical separation of Ni from the co-existing elements was investigated by cation exchange and Ni-Spec extraction chromatography. Precipitation behaviour of Ni and the co-existing elements with dimethylglyoxime(DMG) was investigated in ammonium $citrate/ethanol-H_2O$ and tartaric $acid/acetone-H_2O$ in order to purify separated Ni fractions and to prepare $^{59/63}Ni$ source for the radioactivity measurement using a gas proportional counter. Recovery of Ni separated through ion exchange chromatographic separation procedure was $92.1\%$ with relative standard deviation of $0.9\%$. In addition, recovery of Ni with DMG in the tartaric $acid/acetone-H_2O$ was $85.6\%$ with relative standard deviation of $1.9\%$.

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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.