• 제목/요약/키워드: Anion-exchange HPLC

검색결과 62건 처리시간 0.03초

셀레늄이 강화된 이스트에서 셀레늄 단백질의 분리 및 확인 (Separation and identification of selenoproteins in selenium-enriched yeast)

  • 김경미;박용남
    • 분석과학
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    • 제26권6호
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    • pp.357-363
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    • 2013
  • Selenium-containing proteins were separated from selenium-enriched yeast (SEY) using Trizol$^{(R)}$ reagent followed by anion exchange (AE) chromatography. This method is simpler and less time consuming than electrophoresis. Five selenium containing proteins were identified by on-line AE HPLC-ICP/MS (high performance liquid chromatography-inductively coupled plasma/mass spectrometry). Each protein was enzymatically hydrolyzed to seleno-amino acids and separated with RP (reverse phase) HPLC for the identification of selenoproteins.

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

  • 이상기;김기욱;양자열;인상환;이수연
    • 약학회지
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    • 제45권4호
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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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광화학적으로 제조된 Superoxide Radical을 이용한 Superoxide Dismutase를 생산하는 해양미생물의 탐색 및 효소정제 (Screening and Purification of Superoxide Dismutase Producing Marine Bacterium Using Photochemically Generated Superoxide Ion)

  • 조기웅
    • 미생물학회지
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    • 제38권2호
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    • pp.81-85
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    • 2002
  • 해양세균으로부터 superoxide dismutase를 생산하는 균주를 광화학적 superoxide ion 형성 및 nitrite정량방법으로 검색하여 고활성 균주 B-446을 확보하고 본 균주로부터 35-75% ammonium sulfate precipitation, DEAE-Sephadex A-25 ion exchange chromatography, Sephadex C-200 gel filtration chromatography, High-Q anion exchange chromatography그리고 HPLC-GPC를 이용하여 수율 6%, 정제도 32.3 배의 정제된 SOD를 얻었으며 정제 과정중 효소활성 분석에 본 광화학적 superoxide ion생성 방법으로 사용하였다.

고성능 액체크로마토그래피(HPLC)를 이용한 3가, 6가 크롬의 동시정량에 관한 연구 (Simultaneous Determination of Chromium (III) and Chromium(VI) by High Performance Liquid Chromatography(HPLC))

  • 노재훈;김치년;김춘성;김규상
    • 한국산업보건학회지
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    • 제4권2호
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    • pp.189-197
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    • 1994
  • Analytic methods for Cr(VI) level in industrial hygienic field were suggested by the National Institute for Occupational Safety and Health(NIOSH method 7600, 7604). There were growing needs for measurement of Cr(III) and Cr(VI) levels simultaneously. Two analytical methods were suggested to determine Cr(III) and Cr(VI) levels simultaneously. The one is method by using reversed phase high peformance liquid chromatography(HPLC) and the other is by using ion exchange HPLC. The purpose of this work was to evaluate the usefulness of these two analytic methods. For the difference of ionic charges of Cr(III)-ethylendiamine tetraacetic acid(EDTA) chelate and $CrO_4{^-2}$, we could detect them simultaneously by ion exchange HPLC. Also, we attempted to determine the levels of Cr(III) and Cr(VI) chelated with sodium diethyldithiocarbamate(NaDDTC) by using reversed phase HPLC. The confirmation of Cr(III) and Cr(VI) were checked by fraction collector and nameless atomic absorption spectrometer. The optimal conditions for the formation of Cr(III)-EDTA chelate were two hours incubation period with pH 5. Cr(III)-EDTA and Cr(VI) in EDTA solution were successfully separated by anion exchange column using $Na_2CO_3/NaOH$ mixture as mobile phase. Peaks of Cr(III)-EDTA and Cr(VI) in EDTA were identified at 5 minutes and 7 minutes of retention time respectively by the ion exchange HPLC. The formation of Cr(III)-NaDDTC and Cr(VI)-NaDDTC chelates were twelve hours incubation period. Cr(III)-NaDDTC and Cr(VI)-NaDDTC chelates were separated by reversed phase column using methanol and water mixture as mobile phase. Peaks of Cr(VI)NaDDTC and Cr(III)-NaDDTC chelates were identified at 13 minutes and 26 minutes of retention time respectively by the reversed phase HPLC. Due to reduction of Cr(VI) to Cr(III), it seems to be not suitable for simultaneous determination of Cr(III)-NaDDTC and Cr(VI)-NaDDTC chelates by reversed phase HPLS. Simultaneos determination of Cr(III) and Cr(VI) by ion exchange HPLC was more accurate and simple method.

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산증류-HPLC법과 모니어윌리암스변법을 이용한 절임류중의 이산화황 함량 분석 (Determination of Sulfur Dioxide in Pickles by Acid Distillation-HPLC Method and Monnier Williams Modified Method)

  • 정소영;김일영;김성단;장미라;장민수;한기영
    • 한국식품과학회지
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    • 제35권6호
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    • pp.1028-1032
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    • 2003
  • 절임류에 함유된 이산화황함량의 정확한 분석을 위해 산 증류후 HPLC-UV detector를 이용한 방법과 Monnier-Williams변법으로 실험해보았다. 산증류-HPLC-UV법은 절임류중의 아황산염을 산증류 시킨 후 발생하는 이산화황을 1% triethanolamine용액에 흡수시켜 이를 UV 240nm에서 정량하는 것으로서, 고정상은 음이온 교환컬럼을, 이동상으로는 1.8mM $Na_2Co_3$와 1.7mM $NaHCO_3$용액을 사용하였다. 이 방법으로 단무지에 아황산염을 첨가하여 실험 한 결과 $84.0{\sim}91.7%$의 회수율을 얻었다. 시중에 유통되는 48종의 절임류에 대해 모니어윌리암스변법과 산증류-HPLC-UV법을 이용하여 이산화황 함량을 조사해본 결과, 산증류-HPLC-UV법에 비해 모니어월리암스법에서 휘발성 산으로 인해 높은 농도를 나타내었다. 산증류-HPLC-UV법으로 분석한 절임류의 이산화황 농도는 불검출${\sim}173.05ppm$이었고, 기준인 30ppm을 초과하는 제품은 3건이었다.

Studies on the interaction of thiamines and cyclodextrins

  • Im, In-Seon;Lee, Wang-Kyu;Park, Man-Ki;Kim, Bak-Kwang
    • Archives of Pharmacal Research
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    • 제6권1호
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    • pp.35-44
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    • 1983
  • Interactions between thiamine.HCl and its disulfide derivatives TTFD. TPD and .alpha., .betha. cyclodextrins were investigated. By measuring the H-NMR, C-NMR chemical shifts, the assumption that cyclodextrin may from a inclusion complex with thiamines was supported qualitatively. To calculated the stability constants of them, anion exchange chromatography was applied. The simple, rapid HPLC method was proved to be pertinent thiamine/cyclodextrin system which was chemically unstable and less soluble.

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Enantioseparation of Neutral Compounds on a Quinine Carbamate-Immobilized Zirconia in Reversed-Phase Capillary Electrochromatography

  • Lee, Mun-Rak;Gwon, Ju-Rim;Park, Jung-Hag
    • Bulletin of the Korean Chemical Society
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    • 제31권1호
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    • pp.82-86
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    • 2010
  • Quinine (QN) is a weak anion-exchange type chiral selector and QN-based silica stationary phases have been widely used for enantioseparation of acidic chiral analytes in HPLC and recently in CEC. In this work we report enantioseparation of non-acidic chiral analytes on a quinine carbamate-immobilized zirconia (QNZ) in reversed-phase (RP) CEC. Influences of pH, composition of the buffer, acetonitrile content and the applied voltage on enantioseparation were examined. Enantiomers of the analytes investigated are well separated in acetonitrile/phosphate buffer mobile phases. Separation data on QNZ were compared to those on QN-bonded silica (QNS). Retention was longer but better enantioselectivity and resolution were obtained on QNZ than QNS.

Isolation and Characterization of Proteoglycan Derived From Human Placenta and its Biological Activities

  • Lee, Kyung-Bok;Kim, Jong-Sig;Yoo, Yung-Choon;Kwak, Sang-Tae;Song, Kyung-Sik;Kim, Yeong-Shik
    • Archives of Pharmacal Research
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    • 제23권2호
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    • pp.182-186
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    • 2000
  • Chondroitin sulfates proteoglycans were isolated from human placenta. For the identification of enzymatic digestion products of isolated proteoglycan, strong anion exchange-high performance liquid chromatography (SAX-HPLC) was performed. By the action of chondroitin ABC and chondroitin B lyase, three unsaturated disaccharides 2-acetamide-2-deoxy-3-O-($\beta$-D-gluco-4-enepyranosyluronic acid)-D-galactose ($\delta$Di-OS), 2-acetamide-2-deoxy-3-O-($\beta$-D-gluco-4-enepyranosyluronic acid)-6-O-su lfo-D-galactose ($\delta$Di-6S) and 2-acetamide-2-deoxy-3-O-($\beta$-D-gl uco-4-enepyranosyluronic acid)-4-O-sulfo-D-galactose ($\delta$Di-4S) were produced from the human placenta proteoglycan. The anticoagulant activity of chondroitin sulfate proteoglycan was evaluated by activated partial thromboplastin time (aPTT) assay and thrombin time (TT) assay. The clotting times of aPTT and TT were increased from 72 to 144 sec and 19 to 27 sec, respectively. The Immune-modulating activity of chondroitin sulfate proteoglycan was examined by cell proliferation assay and these results suggest that it may play a role in suppression of the function of immune-related cells.

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감자 더뎅이병원균에 대해 길항활성을 갖는 방선균 탐색 및 항균 활성물질의 분리 (Screening of Antagonistic Actinomycetes for Potato Scab Control and Isolation of Antibiotic Compound)

  • 이향범;조종운;임치환;김창진
    • Applied Biological Chemistry
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    • 제47권2호
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    • pp.164-169
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    • 2004
  • 감자더뎅이병 방제용 생물제제(biocontrol agenL BCA)를 개발하기 위하여 국내 토양으로부터 분리된 5,000여 방선균 균주를 대상으로 더뎅이병 관련 병원균(Streptomyces scabiei및 S. turgidiscabies)에 대한 in vitro또는 in vivo 활성검정을 실시하였다. 활성검정 결과 길항력이 우수한 균주로서 9균주가 선발되었으며 실제 재배포장에서 사용되고 있는 dazomet및 mancozeb 등 농약에 대한 감수성 시험을 실시하여 A020645 균주가 길항활성 뿐만 아니라 약제저항성이 가장 높음을 확인하고 더뎅이병 방제용 BCA 균주로 선발하였다. 본 균주로부터 항균활성물질을 분리하기 위하여 액체배양액으로부터 음이온교환 크로마토그래피(anion exchange chromatography), solidphase(ODS) extraction, TLC, 역상 HPLC 등을 실시하여 최종적으로 compound A와 B를 순수 분리하였다. Compound A와 B는 NMR 분석 결과 nucleoside계 화합물로 판단된다.

Methanol 자화 방선균으로부터 Aminoglycoside 내성 저해물질의 정제 및 특성 (Purification and Characterization of Aminoglycoside-Resistant inhibitior from methylotrophic Actinomycetes)

  • 김현수;신재욱
    • 한국미생물·생명공학회지
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    • 제27권3호
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    • pp.215-222
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    • 1999
  • Methylotrophic actinomycetes No. 155 produced an aminoglycoside antibiotics(AG)-resistant inhibitor. We have previously reported that the inhibitor shows strong inhibition to sisomicin-resistant strain. In order to understand the functions of inhibitor and sisomicin-resistance, characterizations and purification of inhibitor were investigated. Strain No. 155 was tentatively identified as Nocardiopsis sp. based on morphological and some physiological characteristics. In the antimicrobial activity test, the addition of inhibitor to sisomicin showed a reduction effect of MIC on the test strains such as Gram(+), Gram(-) bacteria and yeasts. The combination of the inhibitor and various antibiotics revealed synergistic against E. coli K-12 and B. subtilis PCI 219. The induced intracellular proteins from sisomicin-resistant strain exhibited the sisomicin inactivation by invitro test. And the induced intracellular proteins were inactivated by addition of the inhibitor. The inhibitor compound was purified by anion exchange chromatography(Dowex-1) and HPLC using Asahipak ES-502C column. The purified inhibitor compound was detected in a single peak(above 98.5% purity) through the HPLC analysis.

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