• Title/Summary/Keyword: Gel chromatography

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Studies on screening test for carbamate insecticides by thin layer chromatography (Thin Layer Chromatography에 의한 유기 카바메이트계 농약의 스크리닝 테스트에 관한 연구(1))

  • 박성우
    • Journal of Environmental Health Sciences
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    • v.6 no.1
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    • pp.23-26
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    • 1979
  • 한국에서 사용하고 있는 carbamate계 살충제의 사용율(%)은 1968년경에는 2.1%인데 비해서 현재는 급증되고 있는 실정에 있다. 따라서 이로 인한 자타살 사건, 환경오염, 살포시 급만성 중독 등의 사고가 빈번히 발생하고 있는 동시에 법화학적 분석에도 많은 문제점이 되두되고 있으므로 1차적으로 시판되고 있는 carbamate계 살충제를 thin layer chromatography에 의해서 성분의 분리 확인 시험을 실시한 결과 1) Silica Gel G-60 및 Silica Gel F-254에서 적합하였고 2) 전개용매는 chloroform: benzeneP: carbontetra chloride (60:30:25), cyclohexane: acetone (80:20), cyclohexane:acetone(70:30), ether:hexane(80:20)에서 9종의 살충제가 잘 분리되었으며 3) UV light (2537 ${\AA}$) 및 2.6-dibromoquinone-4-chlorimide는 전개는 Rf치를 육안으로 구분하는데 가장 양호하였고 4) Carbamate계 살충제 중 Bux-2와 Sevin은 Silica Gel G-60에서 UV light (2537 ${\AA}$)에서 특이한 청보라 및 청색형광을 관찰할 수 있었다. 5) 이상의 결과를 보아 법화학적 시료중에 함유된 카바메이트계 농약의 분석은 가능할 것이며 또한 이 방법의 적용이 현재의 실정으로서는 적합한 것으로 사료되는 바이다.

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Purification of Eicosapentaenoic Acid (EPA) and Docosahexaenoic Acid (DHA) Esters from Squid Oil by Silver Ion Chromatography (은 이온 크로마토그래피에 의한 오징어유로부터 eicosapentaenoic acid(EPA) 및 docosahexaenoic acid(DHA)의 분리농축)

  • Gyoung, Young-Soo;Yu, Ying-Lian;Yoon, Jung-Ro
    • Korean Journal of Food Science and Technology
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    • v.36 no.2
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    • pp.361-364
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    • 2004
  • EPA and DHA extracted from methyl esterified squid oil were purified by silver exchanged resin, silver nitrate-impregnated silica gel, silver exchanged zeolite, and silica gel column chromatography, among which column chromatography using mixture of silver exchanged resin and silica gel (10% by weight) showed the best result. By this simple purification method, EPA and DHA were concentrated from 12.5 to 27.9% (yield, 86,0%) and from 21.7 to 49.5% (yield, 87.3%), respectively. Silver exchanged resin had additional advantages of outstanding reusability and simple recovery of silver.

Investigation of brassinosteroid substances in the shoots of japonica type Korean rice (일반계 벼의 brassinosteroid활성물질 검색)

  • Hyun, Kyu-Hawn;Park, Keun-Hyung
    • Applied Biological Chemistry
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    • v.33 no.4
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    • pp.307-314
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    • 1990
  • Brassinosteroid-like substances in two japonica type Korean rices were investigated. The extracts from the shoots at the maximum tillering stage were purified by solvent fractionation, silica gel adsorption chromatography, Sephadex LH-20 chromatography, charcoal adsorption chromatography, preparative TLC, Bondesil chromatography and HPLC of normal phase and reverse phase, successively. Biological activity of each purification step were monitored by the rice lamina inclination test. Two cultivars tested in this experiment produced brassinosteroids and endogenous brassinosteroids showing similiarity between two cultivars.

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Separation and Purification of Angiotensin-I Converting Enzyme Inhibitory Peptides from Layer Hydrolysate (김 가수분해물로부터 Angiotensin-I Converting Enzyme저해 Peptide의 분리$\cdot$정제)

  • LEE Heon-Ok;KIM Dong-Soo;DO Jeong-Ryong;KWAN Dae-Young
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.34 no.2
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    • pp.164-172
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    • 2001
  • The angiotensin-I converting enzyme (ACE) inhibitors from laver hydrolysate was isolated. Among the 13 kinds of proteases, Maxazyme NNP was most effective for preparing the high ACE inhibitory compound. In extraction conditions of ACE inhibitory peptide from laver hydrolysate, ACE inhibitory activity of hydrolysate treated with diethylether for decolorization and that of $70\%$ ethanol soluble fraction among the different ethanol concentrations were higher than other preparations. Low molecular fraction less than 3,000 dalton of layer hydrolysate separated by ultrafiltration had the highest ACE inhibitory activity, for further separation of ACE inhibitory peptide from laver hydrolysate, gel filtration chromatography (Sephadex G-25), reverse-phase HPLC (ODS & Vydac C-18) and gel permeation chromatography (Superdex Peptide HR) were performed. The molecular mass of the ACE inhibitory peptide fractions of gel permeation chromatography determined by electrospray-mass spectrometer were 413.48 (S1O2V2V1P),346.86 (S1O2V2V2P) and 320.32 (S2O6V3V1P) dalton and their amino acid sequence were Val-Gln-Gly-Asn, Thr-Glu-Thr and Phe-Arg, respectively.

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Purification and Characterization of a Juvenile Hormong Binding Protein from Whole Body Homogenates of the Wax Moth, Galleris mellonella Final Instar Larvae (꿀벌부채명나방 종령유충에서 유약호르몬 결합단백질의 정제와 특성)

  • 안기흥;전상학;이경로
    • Korean journal of applied entomology
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    • v.37 no.1
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    • pp.59-64
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    • 1998
  • A juvenile hormone binding protein (JHBP) has been isolated from the whole body homogenate of Galleria rnellonella final instar larvae by gel filtration. The isolated protein is homogenous as judged by column chromatography and gel electrophoresis in the presence and absence of denaturing agent. The JHBP has a relative molecular weight of 32 k by denaturing gel electrophresis and 28 k by gel filtration. The protein exhibits a dissociation constant of 3.9 x M for JH 111.

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Development of Chromatographic Downstream Processing for The Purification of Monoclonal Antibody from Ascites Fluid: Part 1. Tandem Use of Hydroxylapatite Chromatography and Gel Permeation Chromatography (복수로부터의 단세포군 항체의 정제를 위한 크로마토그래프 분리 정제 시스템의 개발 - 1. 히드록실 아파타이트 크로마토그라프와 겔 여과 크로마토그라프로 이루어진 2단계 연속공정 사용 -)

  • Ahn, I.S.;Park, C.Y.
    • Microbiology and Biotechnology Letters
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    • v.17 no.1
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    • pp.19-23
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    • 1989
  • A sequential system composed of hydroxylapatite chromatography and gel permeation chromatography was developed to purify the IgM type monoclonal antibody against the colon cancer cell SC-1 from the ascitic fluid of mice injected with the murine hybridoma CH07E02. In the hydroxylapatite chromatographic step the band dilution could be reduced by controlling the gradient and flow rate of the eluent, the sodium phospate buffer, the optimum values for these variables being 5.82$\times$10$^{-3}$M/cm and 0.2$m\ell/\textrm{cm}^2$/min, respectively. A degree of purity better than 99.99% as judged from silverstaining of the SDS-PAGE bands, was obtained by adding the gel permeation chromatographic step in tandem.

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Chromatographic Fractionation of Protease Inhibitors from Fish Eggs (어류 알로부터 Protease Inhibitors의 크로마토그래피법에 의한 분획)

  • Kim, Jin-Soo;Kim, Ki Hyun;Kim, Hyeon Jeong;Kim, Min Ji;Park, Sung Hwan;Lee, Hyun Ji;Heu, Min Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.46 no.4
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    • pp.351-358
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    • 2013
  • A protease inhibitor from fish eggs was fractionated using chromatographic methods. The fractionation efficiency was evaluated in terms of specific inhibitory activity (SIA, U/mg), purity (fold), total inhibitory activity (TIA, U), and recovery (%). The protease inhibitor (PI) from egg extracts of skipjack tuna (ST Katsuwonus pelamis), yellowfin tuna (YT Thunnus albacares) and Alaska pollock (AP Theragra chalcogramma) was fractionated using Sephadex G-50 gel filtration and DEAE-Sepharose CL-6B anion exchange chromatography based on protein size exclusion and net charge, respectively. Fractions exhibiting strong inhibitory activity were contained in the 30-50 kDa fraction on gel filtration and in the range of 0.4-0.7 M NaCl gradient fraction on anion exchange chromatography. The respective TIA and percent recovery of the fraction obtained with gel filtration toward trypsin and $N{\alpha}$-benzoyl-L-arginine-p-nitroanilide (BAPNA) were 2,758.7 U and 29.6% for ST, 1,005.5 U and 25.6% for YT, and 1,267.5 U and 26.0% for AP. Gel filtration chromatography was more effective at fractionating PI than using ion exchange chromatography. These results suggest that fish eggs act as serine protease inhibitors and might be useful for protease inhibition in foodstuffs.

Studies on the Toxic Activity of Bacillus sphaericus Spo -D1216 Asporogenic Mutant (Bacillus sphaericus Asporogenic Mutant Spo - D1216의 독성에 관한 연구)

  • 복거중;김영한;이형환
    • Microbiology and Biotechnology Letters
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    • v.13 no.2
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    • pp.157-162
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    • 1985
  • The cell cultures and crude extracts of Bacillus sphaericus 1593 K-5 and its mutant Spo-Dl216 were respectively bioassayed against Culex pipiens var. pollens mosquito larvae. The B. sphaeriucs 1593 K-5 showed toxic activity against the larvae. LC$_{50}$ values (cells/$m\ell$) was 2.6$\times$10$^2$. Also the LC$_{50}$ ($\mu\textrm{g}$ Protein/$m\ell$) of the crude extract was 10.26. However, B. sphaericus Spo-Dl216 didn't show toxic activity against the larvae. The soluble cytoplasmic toxin in broken B. sphaeriucs 1593k-5 cells was partially purified by gel permeation chromatography and ion exchange chromatography. Among the fractions of the gel permeation chromatography only a single fraction was found to be toxic. LC$_{50}$ values ($\mu\textrm{g}$ protein/$m\ell$) of the active fraction was 0.182. The active fraction of the gel permeation was subjected to ion exchange chromatography. Only a single fraction showed toxic activity and its LC$_{50}$ values ($\mu\textrm{g}$ protein/$m\ell$) was 0.02..02.

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Purification of Glucoamylase Produced by Rhizopus oryzae (Rhizopus oryzae가 생산(生産)하는 Glucoamylase의 정제(精製))

  • Hou, Won-Nyong;Chung, Man-Jae
    • Korean Journal of Food Science and Technology
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    • v.16 no.3
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    • pp.322-328
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    • 1984
  • These experiments were conducted to purify the glucoamylase produced by Rhizopus oryzae. Two forms of glucoamylase (GI and GII) from Phizopus oryzae were purified by $(NH_2)_2SO_4$ fractionation, acetone fractionation and successive column chromatography on DEAE-cellulose and CM-cellulose. The specific activities of GI and GII toward soluble starch were 157.6 U/㎎. protein (37.5 fold of crude extract), and 164.7 U/㎎. protein (39.2 fold of curde extract), respectively, and the yields of them were 4.3% and 3.8%, respectively. The two purified enzymes have shown a single band by polyacrylamide disc gel electrophoresis and SDS-polyacrylamide gel electrophoresis. The protein bands of their electrophoresis gel were revealed to have glucoamylase activity by iodine staining and were proved to be glycoprotein by periodic acid Schiff's staining.

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Characterization of 65 kD Protein in Latex Excreted from Euphorbia lathyris (Euphorbia lathyris에서 분비되는 Latex 65kD 단백질의 특성규명)

  • Park, Hee-Sung
    • Journal of Plant Biotechnology
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    • v.31 no.4
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    • pp.319-323
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    • 2004
  • Soluble latex protein fraction excreted from Euporbia lathyris laticifer was resolved by 10% SDS-polyacrylamide gel electrophoresis to identify distinctively displayed latex major protein bands including ELp65, ELp55, ELp43, ELp32 and ELp23. Among them, ELp65 was purified by ammonium sulfate precipitation, gel permeation chromatography and ion exchange chromatography. Its N-terminal amino acid sequencing revealed its homology to the leading region of mature peptide of tomato p69a subtilisin-like protease, suggesting a certain role involved in plant defense system. In the analysis of Southern blot hybridization using PCR-amplified tomato p69a probe DNA, E. lathyris genome was suggested to have a gene family consisting of 3-5 gene members putatively encoding subtilisin-like proteases.