Interlaboratory evaluation of multiple LC-MS/MS methods and a commercial ELISA method for determination of tetrodotoxin in oysters and mussels

Andrew D. Turner*, Karl J. Dean, Monika Dhanji-Rapkova, Sonia Dall'Ara, Florella Pino, Claire McVey, Simon Haughey, Natasha Logan, Christopher Elliott, Ana Gago-Martinez, Jose Manuel Leao, Jorge Giraldez, Ryan Gibbs, Krista Thomas, Ruth Perez-Calderon, Dermot Faulkner, Hugh McEneny, Veronique Savar, Damien Reveillon, Philipp HessFabiola Arevalo, J. Pablo Lamas, Eva Cagide, Mercedes Alvarez, Alvaro Antelo, Mirjam D. Klijnstra, Michalina Oplatowska-Stachowiak, Tim Kleintjens, Nermin Sajic, Michael J. Boundy, Benjamin H. Maskrey, D. Tim Harwood, Jesús M González Jartín, Amparo Alfonso, Luis Botana

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)
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Abstract

Background
Given the recent detection of tetrodotoxin (TTX) in bivalve molluscs but the absence of a full collaborative validation study for TTX determination in a large number of shellfish samples, interlaboratory assessment of method performance was required to better understand current capabilities for accurate and reproducible TTX quantitation using chemical and immunoassay methods.

Objective
The aim was to conduct an interlaboratory study with multiple laboratories, using results to assess method performance and acceptability of different TTX testing methods.

Methods
Homogenous and stable mussel and oyster materials were assessed by participants using a range of published and in-house detection methods to determine mean TTX concentrations. Data were used to calculate recoveries, repeatability, and reproducibility, together with participant acceptability z-scores.

Results
Method performance characteristics were good, showing excellent sensitivity, recovery, and repeatability. Acceptable reproducibility was evidenced by HorRat values for all LC–MS/MS and ELISA methods being less than the 2.0 limit of acceptability. Method differences between the LC–MS/MS participants did not result in statistically different results. Method performance characteristics compared well with previously published single-laboratory validated methods and no statistical difference was found in results returned by ELISA in comparison with LC–MS/MS.

Conclusion
The results from this study demonstrate that current LC–MS/MS methods and ELISA are on the whole capable of sensitive, accurate, and reproducible TTX quantitation in shellfish. Further work is recommended to expand the number of laboratories testing ELISA and to standardize an LC–MS/MS protocol to further improve interlaboratory precision.

Highlights
Multiple mass spectrometric methods and a commercial ELISA have been successfully assessed through an interlaboratory study, demonstrating excellent performance.

Original languageEnglish
Pages (from-to)356-369
Number of pages14
JournalJournal of AOAC International
Volume106
Issue number2
Early online date06 Jan 2023
DOIs
Publication statusPublished - Mar 2023

Keywords

  • Agronomy and Crop Science
  • Analytical Chemistry
  • Environmental Chemistry
  • Food Science
  • Pharmacology

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