Analytical and Bioanalytical Chemistry

, Volume 396, Issue 6, pp 2321–2330 | Cite as

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

  • Mònica Campàs
  • Pablo de la Iglesia
  • Margarita Fernández-Tejedor
  • Jorge Diogène
Original Paper

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.

Figure

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

Keywords

Yessotoxin (YTX) Phosphodiesterase (PDE) Inhibition assay Protoceratium reticulatum Liquid chromatography–tandem mass spectrometry (LC-MS/MS) 

Supplementary material

216_2009_3445_MOESM1_ESM.pdf (96 kb)
ESM 1(PDF 96 kb)

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Mònica Campàs
    • 1
  • Pablo de la Iglesia
    • 1
  • Margarita Fernández-Tejedor
    • 1
  • Jorge Diogène
    • 1
  1. 1.IRTASant Carles de la RàpitaSpain

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