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Method for Simultaneous Determination of Anatoxin-a and Microcystins in Korean Water Systems by Using LC/MS/MS  

Moon, Jeong-Suk (Han River Water Environment Research Center, National Institute of Environmental Research)
Kim, Hwa-Bin (Woongjin Coway Environmental Technology Research Center)
Park, Hae-Kyung (Water Environment Research Department)
Lee, Jae-Jung (Geum River Water Environment Research Center)
Park, Jong-Hwan (Youngsan River Water Environment Research Center)
Lee, Hae-Jin (Nakdong River Water Environment Research Center)
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Abstract
This study was purposed to develop an effective LC/MS/MS method for simultaneously determining five pre-treated cyanotoxins (anatoxin-a, microcystins-RR, -YR, -LR and -LA) of cyanobacteria blooms. Cyanobacterial bloom samples were collected from 11 major lakes and three downstream areas of river around Korea during 2005~2009. Cyanotoxins were identified in 38 samples from the lakes. The validity of the method was evaluated and the recovery rates were found ranging from 83~87%. The MDL turned out to be $0.046\;{\mu}g\;L^{-1}$ for anatoxin-a and $0.066\;{\mu}g\;L^{-1}$ for microcystins (RR, YR, LR and LA), which indicates that the method has high sensitivity and accuracy. The most dominant genus of the cyanobacterial blooms was Microcystis, which accounted for 71% of the analysed samples. Microcystis also contained the largest amount of microcystins ($398.5\;{\mu}g\;gDW^{-1}$) among the analyzed cyanobacteria. The analysis of the five cyanotoxins showed that anatoxin-a ranged between $0{\sim}41.833\;{\mu}g\;gDW^{-1}$ and microcystins ranged between $6.311{\sim}2,148.786\;{\mu}g\;gDW^{-1}$. Among the microcystins, micocystin-RR took up 58.3%, the largest portion. Anatoxin-a was found to account for 77.8% of the samples. This study has its significance in that it allowed the establishment of toxin criteria appropriate for the Korean water systems. Further studies may be necessary to conduct for improving water treatment methods.
Keywords
simultaneous determination; LC/MS/MS; microcystin; anatoxin-a; cyanobacteria;
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1 Park, H.D., B.C. Kim, E.K. Kim and T. Okino. 1997. Hepatotoxic microcystines and neurotoxic anatoxina in cyanobacterial blooms from Korean lakes. Environmental Toxicology and Water Quality 13: 225-234.
2 Rapala, J., K. Sivonen, R. Lukkainen and S.I. Niemela. 1993. Anatoxin-a concentration in Anabaena and Aphanizomenon under different environmental conditions and comparason of growth by toxic and non toxic Anabaena strains a laboratory study. Journal of Applied Phycology 5: 581-591.   DOI   ScienceOn
3 Rinehart, K.L., M. Namikoshi and B.W. Choi. 1994. Structure and biosynthesis of toxins from bluegreen algae (Cyanobacteria). Journal of Applied Phycology 6: 159-176.   DOI   ScienceOn
4 Sivonen, K. 2009. Cyanobacterial Toxins. Encyclopedia of Microbiology (Third Edition). pp. 12-24.
5 Sivonen, K., K. Himberg, R. Luukkainen, S.I. Niemela, G.K. Poon and G.A. Codd. 1989a. Preliminary characterization of neurotoxic blooms and strains from Finland. Toxicity Assessment 4: 339-352.   DOI
6 Sivonen, K., W.W. Carmichael, M. Namikoshi, K.L. Rinehart, A.M. Dahlem and S.I. Niemela. 1990. Isolation and characterization of hepatotoxic microcystin homologos from the filamentous freshwater cyanobacterium Nostoc sp. strain 152. Applied and Environmental Microbialogy 56: 2650-2657.
7 Henriksen, P. 1996b. Toxic cyanobacteria/blue-green algae in Danish fresh waters. Thesis, Dept. of Phycology. University of Copenhagen.
8 Hilborn, E.D., W.W. Carmichael, M. Yuan and S.M. Azevedo. 2005. A simple colorimetric method to detect biological evidence of human exposure to microcystins. Toxicon 46(2): 218-221.   DOI   ScienceOn
9 Kim, B.C., E.K. Kim, D.J. Pyo, H.D. Park and W.M. Heo. 1995. Toxic cyanobacterial blooms in Korean lakes. Journal of Korean Society on Water Quality 11(3): 231-237.
10 Kim, J.H., M.A. Yun and H.C. Kim. 2009. Method for simultaneous determination of cyanobacteria in water by LC/MS/MS. Journal of Korean Society on Water Quality 25(4): 597-605.
11 Krishnamyrthy, T., L. Szafraniec, D.F. Hunt, J. Shabanowitz, J.R. Yates, C.R. Hauer, W.W. Carmichael, O. Skulberg, G.A. Codd and S. Missler. 1989. Structural characterization of toxic cyclic peptides from blue-green algae by tandem mass spectrometry. Proceedings of National Academy of Sciences USA 86: 770-774.   DOI   ScienceOn
12 Krishnamurthy, T., W.W. Carmichael and E.W. Sarver. 1986. Investigations of freshwater cyanobacteria (blue-green algae) toxic peptides. I. Isolation, purification and characterization of peptides from Microcystis aeruginosa and Anabaena flos-aque. Toxicon 24: 865-873.   DOI   ScienceOn
13 Ministry of Environment. 2008. Harful Algal Bloom Alert System operating manual.
14 Kusumi, T., T. Ooi, M.M. Watanabe, H. Takahashi and H. Kakisawa. 1987. Cyanoviridin RR, a toxin from the cyanobacterium (blue-green alga) Microcystis viridis. Tetrahedron Letters 28: 4695-4698.   DOI   ScienceOn
15 Masahiko, T., D. Shigeki and Y. Kenji. 1999. Analysis of anatoxin-a infreshwaters by automated on-line derivatization-liquid chromatography-electrospray mass spectrometry. Journal of Chromatography A 862: 191-197.   DOI
16 Meriluoto, J.A., O.A. Sandstrom, J.E. Eriksson, G. Remand, A.G. Graig and J. Chattopadhyaya. 1989. Structure and toxicity of a peptide hepatotoxin from the cyanobacterium Oscillatoria agardhii. Toxicon 24: 1021-1034.
17 Namikoshi, M. and K.L. Rinehart. 1996. Bioactive compounds produced by cyanobacteria. Journal of Industrial Microbialogy & Biotechnology 17: 373-384.   DOI   ScienceOn
18 Codd, G.A. 2000. Cyanobacterial toxins, the perception of water quality, and the prioritisation of eutrophication control. Ecological Engineering 16: 51-60.
19 Dai, M., P. Xie, G. Liang, J. Chen and H. Lei. 2008. Simultaneous dertermination of microcystin-LR and its glutathione conjugate in fish tissues by liquid chromatography tandem mass spectrometry. Journal of Chromatography B 862(1-2): 43-50.   DOI
20 Edwards, C., K.A. Beattie, C.M. Scrimgeour and G.A. Codd. 1992. Identification of anatoxin-a in benthic cyanobacteria (blue-green algae) and in associated dog poisonings at Loch Insh. Scotland. Toxicon 30: 1165-1175.   DOI   ScienceOn
21 Falconer, I., J. Bartram, I. Chorus, T. Kuiper-Goodman, H. Utkilen, M. Burch and J. Codd. 1999. Safe levels and safe practices. p. 155-178. In: Toxic Cyanobacteria in Water a Guide to Their Public Health Consequences, Monitoring and Management (Chorus, I. and J. Bartram, eds.). Spoon Press, on behalf of WHO, London.
22 Falconer, I.R. 1993. Mechanism of peptide toxins from blue-green algae, p. 176-186. In: Algal Toxins in Seafood and Drinking Water (Falconor, I.R. ed.). Academic Press, San Diego.
23 Falconer, I.R. 1999. An overview of problems caused by toxic blue-green algae (cyanobacteria) in drinking and recreational water. Environmental Toxicology 14: 5-12.   DOI
24 Hawkins, P.R., S. Novic, P. Cox, B.A. Neilan, B.P. Burns, G. Shaw, W. Wickramasinghe, Y. Peerapornpisal, W. Euangyuttikarn, T. Itayama, T. Saitou, M. Mizuochi and Y. Inamori. 2005. A review of analytical methods for screening the public health risk from microcystin in the aquatic environment. Journal Water Supply: Research and Technology-AQUA 54.8, 509-518.
25 Harada, K., K. Matsuura, M. Suzuki, H. Oka, M.F. Watanabe, S. Oishi, A.M. Dahlem, V.R. Beasley and W.W. Carmichael. 1988. Analysis and purification of toxic peptides from cyanobacteria by reversed-phase high-performance liquid chromatography. Journal of Chromatography A 448: 275-283.   DOI
26 Harada, K., Y. Kimura, K. Ogawa, M. Suzuki, A.M. Dahlem, V.R. Beasley and W.W. Carmichael. 1989. A new procedure for the analysis and purification of naturally occurring anatoxin-a from the bluegreen algae Anabaena flos-aquae. Toxicon 27: 1289-1296.   DOI   ScienceOn
27 Harada, K.I., I. Ohtani, K. Iwamoto, M. Suzuki, M.F. Watanabe, M. Watanabe and K. Terao. 1994. Isolation of cylindrospermopsin from a cyanobacterium Umezakia natans and its screening method. Toxicon 32: 73-84.   DOI   ScienceOn
28 Baker, P.D. and A.R. Humpage. 1994. Toxicity associated with commonly occurring cyanobacteria in surface waters of the Murray-Darling Basin. Austrailia. Journal of Marine and Freshwater Research 45: 773-786.   DOI
29 Bruno, M., D.A. Barbini, E. Pierdominici, A.P. Serse and A. Ioppolo. 1994. Anatoxin-a and previously unknown toxin in Anabaena planctonica from blooms found in Lake Mulargia (Italy). Toxicon 32: 369-373.   DOI   ScienceOn
30 Botana, L. 2007. Phycotoxins, Chemistry and Biochemistry. Blackwell Publishing. ISBN-13:978-0-8138-2700-1. 345pp.
31 Bumke-Vogt, C., W. Mailahn and I. Chorus. 1999. Anatoxin-a and neurotoxic cyanobacteria in German lakes and reservoirs. Environmental Toxicology 14: 117-125.   DOI
32 Carmichael, W.W. 1992. Cyanobacteria sceondary metabolites the cyanotoxins. Journal of Applied Bacteriology 72(6): 445-459.   DOI
33 Carmichael, W.W., D.F. Biggs and P.R. Gorham. 1975. Toxicology and pharmacological action of Anabaena flos-aquae toxin. Science 187: 542-544.   DOI
34 Carmichael, W.W. and I.R. Falconer. 1993. Diseases related to freshwater blue-green algal toxins, and control measures, p. 187-209. In: Algal Toxins in Seafood and Drinking Water (Falconer, I.R. ed.). Academic Press, London.
35 Chorus, I. and J. Bartram. 1999. Toxic Cyanobacteria in Water. Spon Press, London.