Environmental Engineering Research

  • 제14권4호
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  • Pages.250-254
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  • 2009
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  • 1226-1025(pISSN)
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  • 2005-968X(eISSN)
대한환경공학회 (Korean Society of Environmental Engineers)
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Evaluation of Methods for Cyanobacterial Cell Lysis and Toxin (Microcystin-LR) Extraction Using Chromatographic and Mass Spectrometric Analyses

  • Kim, In S. (Bio-Environmental Engineering Laboratory, Department of Environmental Science and Engineering, Gwangju Institute of Science and Technology) ;
  • Nguyen, Giang-Huong (Bio-Environmental Engineering Laboratory, Department of Environmental Science and Engineering, Gwangju Institute of Science and Technology) ;
  • Kim, Sung-Youn (Bio-Environmental Engineering Laboratory, Department of Environmental Science and Engineering, Gwangju Institute of Science and Technology) ;
  • Lee, Jin-Wook (Bio-Environmental Engineering Laboratory, Department of Environmental Science and Engineering, Gwangju Institute of Science and Technology) ;
  • Yu, Hye-Weon (Bio-Environmental Engineering Laboratory, Department of Environmental Science and Engineering, Gwangju Institute of Science and Technology)
  • 발행 : 2009.12.31
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초록

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.

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참고문헌

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피인용 문헌

  1. Chemical lysis of cyanobacteria vol.9, pp.1, 2015, https://doi.org/10.1186/s13036-015-0007-y
  2. Histopathological and biochemical effects of cyanobacterial cells containing microcystin-LR on Tilapia fish vol.30, pp.1-2, 2016, https://doi.org/10.1111/wej.12169
  3. Phylogenomic Analysis of the Microviridin Biosynthetic Pathway Coupled with Targeted Chemo-Enzymatic Synthesis Yields Potent Protease Inhibitors vol.12, pp.6, 2017, https://doi.org/10.1021/acschembio.7b00124
  4. Electrochemical Flow-ELISA for Rapid and Sensitive Determination of Microcystin-LR Using Automated Sequential Injection System vol.17, pp.7, 2017, https://doi.org/10.3390/s17071639
  5. A high throughput targeted and non-targeted method for the analysis of microcystins and anatoxin-A using on-line solid phase extraction coupled to liquid chromatography–quadrupole time-of-flight high resolution mass spectrometry vol.409, pp.21, 2017, https://doi.org/10.1007/s00216-017-0437-0
  6. The Effect of Cyanobacterial Biomass Enrichment by Centrifugation and GF/C Filtration on Subsequent Microcystin Measurement vol.7, pp.3, 2015, https://doi.org/10.3390/toxins7030821
  7. The Development of an Effective Bacterial Single-Cell Lysis Method Suitable for Whole Genome Amplification in Microfluidic Platforms vol.9, pp.8, 2018, https://doi.org/10.3390/mi9080367
  8. Development of a real-time capacitive biosensor for cyclic cyanotoxic peptides based on Adda-specific antibodies vol.826, pp.None, 2009, https://doi.org/10.1016/j.aca.2014.03.028
  9. Effects of Different Coagulants on Coagulation Process for Removal of Microcystis aeruginosa vol.25, pp.3, 2021, https://doi.org/10.1007/s12205-021-0673-x
  10. Comparative Assessment of Physical and Chemical Cyanobacteria Cell Lysis Methods for Total Microcystin-LR Analysis vol.13, pp.9, 2021, https://doi.org/10.3390/toxins13090596