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Colorimetric and electrochemical phosphodiesterase inhibition assays for yessotoxin detection: development and comparison with LC-MS/MS

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Abstract

This work describes the development and applicability of two functional assays for the detection of yessotoxin (YTX), a polycyclic ether marine toxin produced by dinoflagellates. The assays are based on the interaction between this toxin and the phosphodiesterase (PDE) enzyme and the subsequent measurement of the enzyme activity by colorimetric and electrochemical methods. Firstly, several enzyme substrates were tested in order to select those able to be detected by colorimetry or electrochemistry after enzymatic hydrolysis. The substrates that provided the highest absorbance values and density currents were p-nitrophenyl phenylphosphonate and α-naphthyl phosphate, respectively. After optimisation of the experimental parameters, limits of detection of 0.8 and 0.6 µM were attained by colorimetry and electrochemistry, respectively. An inhibitory effect of YTX on the PDE activity was observed. The assays have been applied to the analysis of YTX production by Protoceratium reticulatum cultures, and results were compared with liquid chromatography–tandem mass spectrometry analysis.

Schematic representation of the electrochemical phosphodiesterase inhibition assay for yessotoxin detection.

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Acknowledgements

This research was funded by the Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria through the projects RTA2006-00103-00-00 and RTA2008-00084-00-00. Dr. Campàs acknowledges financial support from the Departament d’Educació i Universitats de la Generalitat de Catalunya through the Beatriu de Pinós programme and the Ministerio de Ciencia e Innovación and the Fondo Social Europeo through the Ramón y Cajal programme. Technical assistance of IRTA’s staff is gratefully acknowledged.

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  1. IRTA, Ctra. Poble Nou, km. 5.5, 43540, Sant Carles de la Ràpita, Spain

    Mònica Campàs, Pablo de la Iglesia, Margarita Fernández-Tejedor & Jorge Diogène

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  1. Mònica Campàs
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  2. Pablo de la Iglesia
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  3. Margarita Fernández-Tejedor
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  4. Jorge Diogène
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Correspondence to Mònica Campàs.

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Campàs, M., de la Iglesia, P., Fernández-Tejedor, M. et al. Colorimetric and electrochemical phosphodiesterase inhibition assays for yessotoxin detection: development and comparison with LC-MS/MS. Anal Bioanal Chem 396, 2321–2330 (2010). https://doi.org/10.1007/s00216-009-3445-x

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