Detection of free and covalently bound microcystins in animal tissues by liquid chromatography-tandem mass spectrometry |
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Authors: | Milla-Riina Neffling Emilie Lance |
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Institution: | a Department of Biochemistry and Pharmacy, Åbo Akademi University, Tykistökatu 6 A, Biocity 3rd floor, FI-20520, Turku, Finland b UMR CNRS Ecobio 6553, University of Rennes 1, Avenue du Général Leclerc, 35042, Rennes Cedex, France |
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Abstract: | Microcystins are cyanobacterial hepatotoxins capable of accumulation into animal tissues. The toxins act by inhibiting specific protein phosphatases and both non-covalent and covalent interactions occur. The 2-methyl-3-methoxy-4-phenylbutyric acid (MMPB) method determines the total, i.e. the sum of free and protein-bound microcystin in tissues. The aim of the method development in this paper was to tackle the problems with the MMPB methodology: the rather laborious workflow and the loss of material during different steps of the method. In the optimised workflow the oxidation recovery was of acceptable level (29-40%), the extraction efficiency good (62-97%), but the signal suppression effect from the matrix remained severe in our system (16-37% signal left). The extraction efficiency for the determination of the free, extractable microcystins, was found to be good, 52-100%, depending on the sample and the toxin variant and concentration. |
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Keywords: | Microcystins MMPB method Total contamination levels |
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