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Faster ciguatoxin extraction methods for toxicity screening.

Christopher R Loeffler1, Astrid Spielmeyer2

  • 1Department of Safety in the Food Chain, National Reference Laboratory for the Monitoring of Marine Biotoxins, German Federal Institute for Risk Assessment, Max-Dohrn-Str. 8-10, 10589, Berlin, Germany. Christopher.loeffler@bfr.bund.de.

Scientific Reports
|September 17, 2024
PubMed
Summary
This summary is machine-generated.

New methods for extracting ciguatoxins (CTXs) from fish offer faster, more accurate testing for ciguatera poisoning (CP). These high-throughput techniques improve seafood safety and global screening capabilities for CTX contamination.

Keywords:
(max. 6, American spelling): marine biotoxinsCiguateraExtractionFish poisoningMass spectrometryNeuro-2a bioassay

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Area of Science:

  • Food safety science
  • Analytical chemistry
  • Toxicology

Background:

  • Ciguatera poisoning (CP) is a significant global health issue linked to consuming seafood contaminated with ciguatoxins (CTXs).
  • Current methods for detecting CTXs in seafood are hindered by complex extraction and slow cleanup procedures, limiting high-throughput analysis.
  • The increasing global demand for seafood necessitates rapid and reliable testing for CTX contamination.

Purpose of the Study:

  • To develop and compare novel, high-throughput methods for extracting ciguatoxins (CTXs) from various fish tissues.
  • To evaluate the sensitivity, specificity, and robustness of these new extraction protocols.
  • To enhance global capabilities for screening seafood for CTX contamination and mitigate public health risks.

Main Methods:

  • Extraction of CTXs from diverse fish species (wet and freeze-dried) using mechanical maceration (acetone or methanol) or enzymatic digestion followed by solvent extraction.
  • Assessment of crude extracts for CTX-like toxicity using an in vitro mouse neuroblastoma (N2a) cell-based assay (CBA).
  • Confirmation of CTX analog retention using liquid chromatography-tandem mass spectrometry (LC–MS/MS) and comparison with international seafood contaminant guidance.

Main Results:

  • No significant differences in toxin estimates were observed among the three extraction methods (p=0.219), indicating their interchangeability.
  • Methanol-based and enzyme-based extraction methods demonstrated superior speed (>16 samples/day) and 100% accuracy.
  • All tested extraction methods successfully confirmed CTX analogs and met international seafood market contaminant guidance.

Conclusions:

  • Developed extraction methods significantly increase sample processing throughput for CTX analysis.
  • Methanol and enzyme-based extraction protocols are preferred for their speed, accuracy, and reliability in detecting CTXs.
  • These advancements can alleviate bottlenecks in CTX screening, improving environmental and human health protection against ciguatera poisoning.