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Updated: May 21, 2026

Improved Polymerase Chain Reaction-restriction Fragment Length Polymorphism Genotyping of Toxic Pufferfish by Liquid Chromatography/Mass Spectrometry
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A comparative study for PSP toxins quantification by using MBA and HPLC official methods in shellfish.

B Ben-Gigirey1, M L Rodríguez-Velasco, A Otero

  • 1European Union Reference Laboratory for Marine Biotoxins-EURLMB, Spanish Food Safety and Nutrition Agency, Ministry of Health, Social Policy and Equality, CITEXVI, Campus Universitario de Vigo, Fonte das Abelleiras, Lagoas-Marcosende, 36310 Vigo, Spain.

Toxicon : Official Journal of the International Society on Toxinology
|June 12, 2012
PubMed
Summary

This study compared two methods for detecting Paralytic Shellfish Poisoning (PSP) toxins in molluscs. Results showed 85% agreement with legal limits, though some differences in toxicity were observed between methods.

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Area of Science:

  • Marine Biology
  • Food Safety
  • Analytical Chemistry

Background:

  • Commission Regulation (EC) N° 2074/2005 established the biological method as the reference for Paralytic Shellfish Poisoning (PSP) toxins detection.
  • Commission Regulation (EC) N° 1664/2006 introduced the Lawrence method as an alternative to the reference method.

Purpose of the Study:

  • To compare AOAC Official Methods of Analysis 959.08 (Biological method) and 2005.06 (Liquid Chromatography with fluorescence detection) for PSP toxin detection.
  • To evaluate the influence of extraction solvent on total sample toxicity.

Main Methods:

  • Analysis of 40 diverse shellfish samples (mussels, clams, scallops, etc.) using both official methods.
  • Acetic and hydrochloric acid extractions performed on all samples, followed by simultaneous analysis.
  • Comparison of mouse bioassay (MBA) with HCl extraction and Liquid Chromatography with fluorescence detection (HPLC-FLD) with acetic acid extraction.

Main Results:

  • 85% of samples showed consistent results regarding compliance with legal limits between MBA and HPLC-FLD.
  • A linear correlation coefficient of r² = 0.69 was observed, with no significant differences indicated by paired t-tests.
  • Toxicity discrepancies were noted: higher levels by MBA in 15/18 Gymnodinium-profiled shellfish, and higher levels by HPLC-FLD in 9 samples.

Conclusions:

  • Both AOAC methods demonstrate good agreement for regulatory compliance in PSP toxin detection.
  • Differences in toxicity measurements highlight the need for further investigation into extraction, clean-up, and toxin profile complexities.
  • The study underscores potential challenges in accurately quantifying diverse PSP toxin profiles using current official methods.