• Title/Summary/Keyword: Anion-exchange HPLC

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

  • Kim, Kyong-Mi;Pak, Yong-Nam
    • Analytical Science and Technology
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    • v.26 no.6
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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.

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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Screening and Purification of Superoxide Dismutase Producing Marine Bacterium Using Photochemically Generated Superoxide Ion (광화학적으로 제조된 Superoxide Radical을 이용한 Superoxide Dismutase를 생산하는 해양미생물의 탐색 및 효소정제)

  • 조기웅
    • Korean Journal of Microbiology
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    • v.38 no.2
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    • pp.81-85
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    • 2002
  • A marine bacterium producing superoxide dismutase, strain number B446, was screened with nitrite quantitation method using hydroxy amine and photochemically generated superoxide ion, and the superoxide dismutase was purified through 35-75% ammonium sulfate precipitation, DEAE-Sephadex A-25 ion exchange chromatography, Sephadex G-200 gel filtration chromatography, and High-Q anion exchange chromatography to a yield of 6% and purification fold of 32.3.

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

  • Roh, Jae Hoon;Kim, Chi Nyon;Kim, Choon Sung;Kim, Kyoo Sang
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.4 no.2
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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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Determination of Sulfur Dioxide in Pickles by Acid Distillation-HPLC Method and Monnier Williams Modified Method (산증류-HPLC법과 모니어윌리암스변법을 이용한 절임류중의 이산화황 함량 분석)

  • Jung, So-Young;Kim, Il-Young;Kim, Sung-Dan;Jang, Mi-Ra;Chang, Min-Su;Han, Ki-Young
    • Korean Journal of Food Science and Technology
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    • v.35 no.6
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    • pp.1028-1032
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    • 2003
  • To assess accurate methods for measuring sulfur dioxide residue in pickles, the acid distillation HPLC-UV method and Monnier Willams modified method were examined. By the acid distillation HPLC-UV method, sulfites released from pickles by acid distillation were absorbed in 1% triethanolamine solution and detected as sulfite ion by HPLC with UV monitoring at 240nm. An anion exchange column was employed with 1.8mM $Ma_2CO_3-1.7mM\;NaHCO_3$ solution as a mobile phase, $84.0{\sim}91.7%$ of sulfite added to pickled radish were recovered. Total sulfite levels from 48 kinds of pickles analyzed by acid distillation HPLC-UV was compared with those analyzed by the Monnier Williams modified method. The Monnier Williams modified method showed higher levels of sulfur dioxide than the acid distillation HPLC-UV method due to the presence of volatile acids in pickles. The concentration of sulfur dioxide was in the range of $N.D{\sim}173.05ppm$ in pickled radish and over 30ppm of sulfur dioxide from 3 samples by the acid distillation-HPLC-UV method.

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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    • v.6 no.1
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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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    • v.31 no.1
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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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    • v.23 no.2
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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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Screening of Antagonistic Actinomycetes for Potato Scab Control and Isolation of Antibiotic Compound (감자 더뎅이병원균에 대해 길항활성을 갖는 방선균 탐색 및 항균 활성물질의 분리)

  • Lee, Hyang-Burm;Cho, Jong-Wun;Lim, Chi-Hwan;Kim, Chang-Jin
    • Applied Biological Chemistry
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    • v.47 no.2
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    • pp.164-169
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    • 2004
  • In the course of our screening for biocontrol agent (BCA) against Streptomyces scabiei and S. turgidiscabies causing potato scab using 5,000 actinomtcete isolates, 9 antagonistic strains were selected as BCA candidates through in vitro and in vivo assay. An antagonistic strain, A020645 was highly resistant to some pesticides and antibiotics such as dazomet and mancozeb and showed high control value in vivo. Two bioactive compounds (compound A, B) were purified by anion exchange chromatography, solid phase (ODS) extraction, TLC and reverse phase HPLC. Their chemical structures are now thought to be nucleoside derivative as determined by $^1H-NMR$ data analysis. Their full chemical structures would be elucidated through $^{13}C-NMR$, HMQC and HMBC analyses. Further studies will be focused on fitness in soil and formulation of the BCA candidates.

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

  • 김현수;신재욱
    • Microbiology and Biotechnology Letters
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    • v.27 no.3
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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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