• 제목/요약/키워드: Microcystin

검색결과 107건 처리시간 0.029초

Evaluation of Methods for Cyanobacterial Cell Lysis and Toxin (Microcystin-LR) Extraction Using Chromatographic and Mass Spectrometric Analyses

  • Kim, In S.;Nguyen, Giang-Huong;Kim, Sung-Youn;Lee, Jin-Wook;Yu, Hye-Weon
    • Environmental Engineering Research
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    • 제14권4호
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    • pp.250-254
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    • 2009
  • Contamination of microcystins, a family of heptapeptide hepatotoxins, in eutrophic water bodies is a worldwide problem. Due to their poisoning effects on animals and humans, there is a requirement to characterize and quantify all microcystins present in a sample. As microcystins are, for most part, intracellular toxins produced by some genera of cyanobacteria, lysing cyanobacterial cells to release all microcystins is considered an important step. To date, although many cell lysis methods have been used, little work has been conducted comparing the results of those different methods. In this study, various methods for cell lysis and toxin extraction from the cell lysates were investigated, including sonication, bead beating, freeze/thaw, lyophilization and lysing with TritonX-100 surfactant. It was found that lyophilization, followed by extraction with 75% methanol, was the most effective for extracting toxins from Microcystis aeruginosa cells. Another important step prior to the analysis is removing impurities and concentrating the target analyte. For these purposes, a C18 Sep-Pak solid phase extraction cartridge was used, with the percentage of the eluent methanol also evaluated. As a result, methanol percentages higher than 75% appeared to be the best eluting solvent in terms of microcystin-leucine-arginine (MC-LR) recovery efficiency for the further chromatographic and mass spectrometric analyses.

Effect of Chlorination on Removal of Cyanobacterial Microcystins

  • Jung, Jong-Mun;Park, Hong-Ki;Lee, You-Jung;Jung, Eun-Young;Kwon, Ki-Won;Shin, Pan-Se;Joo, Gea-Jae
    • 한국환경과학회지
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    • 제11권11호
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    • pp.1157-1163
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    • 2002
  • The effective removal of microcystins by chlorination was investigated on a laboratory scale. With an initial chl.a concentration of more than 1,000 $\mu\textrm{g}$/ℓ, the required chlorine dose for the effective removal of microcystins from the raw water was more than 8.0 mg/ℓ. Whereas, a chlorine dose of 3.0 mg/ℓcould effectively remove microcystins from raw water containing a chl.a concentration of less than 1,000 $\mu\textrm{g}$/ℓ. The microcystin removal was more effective below pH 8.0, plus the optimum pH range was unrelated to the concentration of toxic algal material. Although chlorination is one of the most effective methods for reducing the toxin from blue-green algae, it causes cell lysis and toxin release. However, it was demonstrated that the released cell lysates and toxins could be effectively removed by a higher dose of the oxidant. The highest removal efficiency of dissolved microcystins(initial concentration: 280 $\mu\textrm{g}$ L$\^$-1/) was with a chlorine dose of 5.0 mg/ℓ.

Hepatotoxicity Induced by Microcystin-LR in Rat

  • Kim, Bum-Seok;Cho, Jae-Woo;Kwon, Hyuk-Nyun;Blank, Ivar;Borisova, Irina;Ejaz, Sohail;Chekarova, Irina;Kwon, Jung-Kee;Lim, Chae-Woong
    • Toxicological Research
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    • 제22권4호
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    • pp.375-380
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    • 2006
  • Microcystin-LR (MC-LR) is a cyanobacterial hepatotoxin mainly produced by Microcystis aeruginosa. The current study examined the effects of a single intraperitoneal dose of MC-LR in rats. Female Sprague-Dawley rats were intraperitoneally injected with MC-LR ($100{\mu}g/kg$ body weight) and they were sacrificed at 0, 20, 40, 80, 160 min, or 12 h after injection. Clinically, animals showed lethargy and had ruffled hair beginning at 40 min post injection. In the gross findings, liver was enlarged and its color was changed into dark red beginning at 40 min post injection. Microscopically, dissociation of centrilobular hepatocytes and hemorrhage was observed in the hepatic central legions and such pathological changes were then extended to the portal regions of liver by time course manner. Interestingly at 80 min after MC-LR injection, the entrapped eosinophilic materials that may be necrotic fragments of dissociated hepatocytes were found in the capillaries of lung and renal glomerulus. Ultrastructurally, microvilli of the hepatocytes were disrupted or lost at all time points. Furthermore, the Disse space and gap junctions were widened beginning at 40 min post injection. These results suggest that liver is the major target organ of MC-LR and isolated hepatocytes by the effects of such hepatotoxin may secondarily reduce the physiological function of lung and kidney.

온라인 시료주입과 UPLC-Orbitrap 질량분석법을 이용한 수질 조류독소의 고속분석방법 개발 및 환경시료적용 (Fast and Accurate Determination of Algal Toxins in Water using Online Preconcentration and UPLC-Orbitrap Mass Spectrometry)

  • 장제헌;김윤석;최재원
    • 한국물환경학회지
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    • 제28권6호
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    • pp.843-850
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    • 2012
  • Due to the fast response to algae bloom issue in drinking water treatment plant, very fast determination methodology for algal toxin is required. In this study, column switching technique based online preconcentration method was combined with high resolution full scan mass spectrometer to save sample preparation time and to obtain fast and accurate result. After parameter optimization of online preconcentration, 1mL filtered sample was directly injected to trap column with switching valve system. Next, target toxins are eluted by 98% acetonitrile and analysed with 150 - 1,100 amu scan range at 50,000 resolving power. Method detection limit (MDL) for microcystin-LR, the most toxic isomer, was 0.1 ng/mL and others such as microcystin-YR, microcystin-RR and nodularin were 0.08, 0.03 and 0.04 ng/mL, respectively. This is the best improved sensitivities with 1mL volume in the literature. Furthermore, due to the use of ultra pressure HPLC (UPLC), the whole method run was completed in 4 min. Real sample applications for 173 sample including 55 surface water and 118 treatment plant samples for raw and treated water could be done within 16 hours. In our calculation, this methodology is roughly 80% faster than the previous manual solid-phase extraction with LC-MS/MS method.

Microcystis 3종(M. aeruginosa, M. ichthyoblabe, M. viridis)의 성장과 microcystins 생성에 대한 온도 및 조도의 영향 (Effects of the Temperature and Light Intensity on the Growth and Microcystin Production of Three Species of Microcystis (M. aeruginosa, M. ichthyoblabe, M. viridis))

  • 이경락;정원화;김진희;김한순
    • 생태와환경
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    • 제43권3호
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    • pp.400-408
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    • 2010
  • 수화가 발생한 영천댐과 안계댐에서 분리한 Microcystis (M. aeruginosa, M. ichthyoblabe, M. viridis) 3종의 온도 및 조도에 대한 성장률과 microcystins 생성 특성을 알아보기 위해 배양실험을 실시하였다. 성장률은 3종 모두 $30^{\circ}C$ 이상에서 최고값을 나타내었으나 microcystins은 $20^{\circ}C$에서 최대로 생성되었으며, 온도가 증가할수록 microcystins 생성은 감소하였다. Microcystis aeruginosa와 M. ichthyoblabe은 $70\;{\mu}mol\;m^{-2}\;s^{-1}$의 광 조건에서 최대성장률을 나타낸 반면, M. viridis은 $35\;{\mu}mol\;m^{-2}\;s^{-1}$의 광 조건에서 최대성장률을 나타내었다. Microcystis aeruginosa, M. ichthyoblabe 및 M. viridis는 각각 $120\;{\mu}mol\;m^{-2}\;s^{-1}$, $70\;{\mu}mol\;m^{-2}\;s^{-1}$ 그리고 $35\;{\mu}mol\;m^{-2}\;s^{-1}$의 광 조건에서 최대의 microcystins를 생성하였다. 온도와 조도에 대한 이들 3종의 microcystins 생성정도와 microcystins 타입은 종간에 뚜렷한 차이를 나타내는 종 특이성을 보였다.

Production of Hepatotoxin by the Cyanobacterium Scytonema sp. Strain BT 23

  • Ashok, Kumar;Singh, D.P.;Tyagi, M.B.;Kumar, Arvind;Prasuna, E.G.;Thakur, J.K.
    • Journal of Microbiology and Biotechnology
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    • 제10권3호
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    • pp.375-380
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    • 2000
  • The preliminary screening of several cyanobacteria, using mice bioassay, reveale the production of a hepatotoxin by the cyanobacterium Scytonema sp. strain BT 23 isolated from soil. An intraperitoneal injection of the crude toxin (LD50 56 mg/kg body wt) from this strain caused the death of the mice within 40 min, and the anmals showed slinical signs of mice within 40 min, and the animals showed clinical signs of hepatotoxicity. The toxin was purified and partially characterized. The active fraction appears to be nonpolar in nature and shows absorption peaks at 240 and 285 nm. The purified toxin had an LD50 of TEX>$100<\mu\textrm{g}/kg$ body wt and the test mice died within 40 min of toxin administration. The toxin-treated mice showed a 1.65-fold increase in liver weight at 40 min and the liver color chnged to dark red due to intrahepatic hemorrhage and pooling of blood. Furthermore, the administration of the toxin to test mice induced a 2.58, 2.63, and 2.30-fold increse in the activity of the serum enzymes alanine aminotransferase, lactate dehydrogenase, and alkaline phosphatase, respectively. Further experiments with the 14C-labeled toxin revealed a maximum accumulation of the toxin in the liver. The clinical symptoms in the mice were similar to those produced by microcystin-L.R. These results suggest that hepatotoxins may also be produced in non bloom-forming planktonic cyanobacteria.

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남조류 독소 정량을 위한 스트립분석법과 HPLC 분석법의 비교 (Comparison of strip analysis and HPLC analysis for the quantitative analysis of cyanobacterial toxin)

  • 표동진;임미연
    • 분석과학
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    • 제28권3호
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    • pp.168-174
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    • 2015
  • 남조류 독소인 마이크로시스틴은 여름철 우리나라 여러 호수들에 존재하여 물고기와 가축 그리고 인간에게 강한 독성을 나타내는 독소이다. 본 논문에서는 이러한 수중 남조류 독소, 마이크로시스틴을 정량분석하는 두가지 방법, 즉 고성능 액체크로마토그래피(HPLC) 분석법과 최근 우리 실험실에서 개발한 스트립분석법의 비교를 시도하였다. 두 분석법의 측정가능농도범위가 많이 달라 HPLC 법으로 먼저 측정한 후 시료를 물로 희석시켜 스트립분석법에 적용하였다. 서로 다른 농도의 마이크로시스틴을 함유한 7가지 물시료들을 사용하여 HPLC분석법과 스트립 분석법으로 남조류 독소 총량을 측정하였다. 그 결과 두 분석법의 정량측정결과가 매우 잘 일치하는 것을 볼 수 있었다. 두 분석법의 상관분석 결과 r 값은 0.99998 이었으며 통계적인 유의성을 나타내는 p 값은 0.00001 이었다.

소양호에서 남조류 독소, 마이크로시틴의 정량 분석 (Quantitative Analysis of Microcystins, Cyanobacterial Toxins in Soyang Lake)

  • 이정애;이소영;표동진
    • 대한화학회지
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    • 제46권6호
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    • pp.535-540
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    • 2002
  • 남조류 독소인 마이크로시틴은 호수에서 매우 미량 존재하기 때문에 이를 정확히 정량 분석하기가 쉽지 않다. 본 연구에서는 소양호에서 채취한 물시료 중에 포함된 남조류 독소, 마이크로시틴을 정량 분석하기 위하여 두 가지 서로 다른 정량 분석법을 시도하였으며 이 둘의 정량 결과를 비교${\cdot}$분석 하였다. 첫째는 고체상 추출과 HPLC(High Performance Liquid Chromatography)를 이용한 분석법을 이용하였으며 둘째는 마이크로시틴의 단일클론항체를 이용한 효소면역흡착분석법(Enzyme-Linked Immunosorbent Assay,ELISA)을 이용하였다.