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Analytical and Bioanalytical Chemistry

, Volume 400, Issue 9, pp 3165–3175 | Cite as

Validation of an accelerated solvent extraction liquid chromatography–tandem mass spectrometry method for Pacific ciguatoxin-1 in fish flesh and comparison with the mouse neuroblastoma assay

  • Jia Jun Wu
  • Yim Ling Mak
  • Margaret B. Murphy
  • James C. W. Lam
  • Wing Hei Chan
  • Mingfu Wang
  • Leo L. Chan
  • Paul K. S. LamEmail author
Original Paper

Abstract

Ciguatera fish poisoning (CFP) is a global foodborne illness caused by consumption of seafood containing ciguatoxins (CTXs) originating from dinoflagellates such as Gambierdiscus toxicus. P-CTX-1 has been suggested to be the most toxic CTX, causing ciguatera at 0.1 μg/kg in the flesh of carnivorous fish. CTXs are structurally complex and difficult to quantify, but there is a need for analytical methods for CFP toxins in coral reef fishes to protect human health. In this paper, we describe a sensitive and rapid extraction method using accelerated solvent extraction combined with high-performance liquid chromatography–tandem mass spectrometry (HPLC-MS/MS) for the detection and quantification of P-CTX-1 in fish flesh. By the use of a more sensitive MS system (5500 QTRAP), the validated method has a limit of quantification (LOQ) of 0.01 μg/kg, linearity correlation coefficients above 0.99 for both solvent- and matrix-based standard solutions as well as matrix spike recoveries ranging from 49% to 85% in 17 coral reef fish species. Compared with previous methods, this method has better overall recovery, extraction efficiency and LOQ. Fish flesh from 12 blue-spotted groupers (Cephalopholis argus) was assessed for the presence of CTXs using HPLC-MS/MS analysis and the commonly used mouse neuroblastoma assay, and the results of the two methods were strongly correlated. This method is capable of detecting low concentrations of P-CTX-1 in fish at levels that are relevant to human health, making it suitable for monitoring of suspected ciguateric fish both in the environment and in the marketplace.

Keywords

Ciguatera fish poisoning Ciguatoxin HPLC-MS/MS Accelerated solvent extraction Mouse neuroblastoma assay 

Notes

Acknowledgements

We thank Ms. Rachel P. P. Wong (School of Biological Science, The University of Hong Kong) for identification of fish species and Asia Pacific Application Support Center of AB Sciex for providing 5500 QTRAP and technical assistance. The work described in this paper was fully supported by a grant from the Research Grants Council of the Hong Kong Special Administrative Region, China (CityU3/CRF/08).

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Jia Jun Wu
    • 1
  • Yim Ling Mak
    • 1
  • Margaret B. Murphy
    • 1
  • James C. W. Lam
    • 2
  • Wing Hei Chan
    • 1
  • Mingfu Wang
    • 3
  • Leo L. Chan
    • 2
  • Paul K. S. Lam
    • 1
    Email author
  1. 1.State Key Laboratory in Marine Pollution and Department of Biology and ChemistryCity University of Hong KongKowloonChina
  2. 2.State Key Laboratory in Marine Pollution and Department of Biology and ChemistryCity University of Hong KongKowloonChina
  3. 3.School of Biological SciencesThe University of Hong KongHong KongChina

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