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A mussel tissue certified reference material for multiple phycotoxins. Part 2: liquid chromatography–mass spectrometry, sample extraction and quantitation procedures

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Abstract

A freeze-dried mussel tissue certified reference material (CRM-FDMT1) containing multiple groups of shellfish toxins has been prepared. Toxin groups present in the material include okadaic acid and the dinophysistoxins, azaspiracids, yessotoxins, pectenotoxins, spirolides and domoic acid. In this work, analytical methods have been examined for the characterisation of the candidate CRM. A comprehensive extraction procedure was developed, which gave good recovery (>98%) for all lipophilic toxins studied. A fast liquid chromatography–mass spectrometry (LC-MS) method was developed that separates the major toxins according to the MS ionisation mode of optimum sensitivity. Matrix effects associated with analysis of these extracts using the developed LC-MS method were assessed. Standard addition and matrix-matched calibration procedures were evaluated to compensate for matrix effects. The methods and approaches will be used for the precise characterisation of the homogeneity and stability of the various toxins in CRM-FDMT1 and for the accurate assignment of certified values. The developed methods also have excellent potential for application in routine regulatory monitoring of shellfish toxins.

LC-MS/MS chromatogram of CRM-FDMT1: a mussel tissue CRM containing multiple groups of phycotoxins

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References

  1. Anonymous (2004) Regulation (EC) No 853/2004 of the European parliament and of the council of 29 April 2004 laying down specific hygiene rules for food of animal origin. Official Journal of the European Union L 139 of 30 April 2004

  2. McCarron P, Emteborg H, Nulty C, Rundberget T, Loader JI, Miles CO, Teipel K, Quilliam MA, Hess P (2011) A mussel tissue certified reference material for multiple toxins. Part 1: design and preparation. Anal Bioanal Chem. doi:10.1007/s00216-011-4786-9

  3. Quilliam MA, Hess P, Dell'Aversano C (2001) Recent developments in the analysis of phycotoxins by liquid chromatography-mass spectrometry. De Koe WJ, Samson RA, van Egmond HP, Gilbert J, Sabino M (Eds) Proceedings of the Xth International IUPAC Symposium on Mycotoxins and Phycotoxins, 21–25 May 2000, Guaruja (Brazil), pp 383–391

  4. McNabb P, Selwood AI, Holland PT, Aasen J, Aune T, Eaglesham G, Hess P, Igarishi M, Quilliam M, Slattery D, van de Riet J, van Egmond HP, Van den Top H, Yasumoto T (2005) Multiresidue method for determination of algal toxins in shellfish: single-laboratory validation and interlaboratory study. J AOAC Int 88:761–772

    CAS  Google Scholar 

  5. Fux E, McMillan D, Bire R, Hess P (2007) Development of an ultra-performance liquid chromatography-mass spectrometry method for the detection of lipophilic marine toxins. J Chromatogr A 1157:273–280

    Article  CAS  Google Scholar 

  6. Stobo LA, Lacaze JP, Scott AC, Gallacher S, Smith EA, Quilliam MA (2005) Liquid chromatography with mass spectrometry—detection of lipophilic shellfish toxins. J AOAC Int 88:1371–1382

    CAS  Google Scholar 

  7. Gerssen A, Mulder PPJ, McElhinney MA, de Boer J (2009) Liquid chromatography–tandem mass spectrometry method for the detection of marine lipophilic toxins under alkaline conditions. J Chromatogr A 1216:1421–1430

    Article  CAS  Google Scholar 

  8. Hess P, Nguyen L, Aasen J, Keogh M, Kilcoyne J, McCarron P, Aune T (2005) Tissue distribution, effects of cooking and parameters affecting the extraction of azaspiracids from mussels, prior to analysis by liquid chromatography coupled to mass spectrometry. Toxicon 46:62–71

    Article  CAS  Google Scholar 

  9. Quilliam MA (1995) Analysis of diarrhetic shellfish poisoning toxins in shellfish tissue by liquid chromatography with fluorometric and mass spectrometric detection. J AOAC Int 78:555–570

    CAS  Google Scholar 

  10. Rodrigues SM, Vale P, Botelho MJ, Gomes SS (2009) Extraction recoveries and stability of diarrhetic shellfish poisoning (DSP) toxins in naturally contaminated samples. Food Addit Contam A 26:229–235

    Article  CAS  Google Scholar 

  11. McCarron P, Emteborg H, Hess P (2007) Freeze-drying for the stabilisation of shellfish toxins in mussel tissue (Mytilus edulis) reference materials. Anal Bioanal Chem 387:2475–2486

    Article  CAS  Google Scholar 

  12. Ito S, Tsukada K (2002) Matrix effect and correction by standard addition in quantitative liquid chromatographic-mass spectrometric analysis of diarrhetic shellfish poisoning toxins. J Chromatogr A 943:39–46

    Article  CAS  Google Scholar 

  13. Fux E, Rode D, Bire R, Hess P (2008) Approaches to the evaluation of matrix effects in the liquid chromatography-mass spectrometry (LC-MS) analysis of three regulated lipophilic toxin groups in mussel matrix (Mytilus edulis). Food Addit Contam A 25:1024–1032

    Article  CAS  Google Scholar 

  14. King R, Bonfiglio R, Fernandez-Metzler C, Miller-Stein C, Olah T (2000) Mechanistic investigation of ionization suppression in electrospray ionization. J Am Soc Mass Spectrom 11:942–950

    Article  CAS  Google Scholar 

  15. These A, Scholz J, Preiss-Weigert A (2009) Sensitive method for the determination of lipophilic marine biotoxins in extracts of mussels and processed shellfish by high-performance liquid chromatography–tandem mass spectrometry based on enrichment by solid-phase extraction. J Chromatogr A 1216:4529–4538

    Article  CAS  Google Scholar 

  16. McElhinney MA (2007) Matrix effects. development of clean-up and LC-techniques contributing towards a reference LCMS method for the analysis of lipophilic marine toxins. MPhil Thesis, Dublin Institute of Technology

  17. Gerssen A, Olst EHW, Mulder PPJ, Boer J (2010) In-house validation of a liquid chromatography tandem mass spectrometry method for the analysis of lipophilic marine toxins in shellfish using matrix-matched calibration. Anal Bioanal Chem 397:3079–3088

    Article  CAS  Google Scholar 

  18. NRC-CNRC (2002) Certified calibration solution for the spiromine toxins, 13-desmethyl spirolide C. Certificate of analysis, Institute for Marine Biosciences, Halifax, NS, Canada

  19. Yasumoto T, Takizawa A (1997) Fluorometric measurement of yessotoxins in shellfish by high-pressure liquid chromatography. Biosci Biotechnol Biochem 61:1775–1777

    Article  CAS  Google Scholar 

  20. Kilcoyne J, Fux E (2010) Strategies for the elimination of matrix effects in the liquid chromatography tandem mass spectrometry analysis of the lipophilic toxins okadaic acid and azaspiracid-1 in molluscan shellfish. J Chromatogr A 1217:7123–7130

    Article  CAS  Google Scholar 

  21. Gerssen A, McElhinney MA, Mulder PPJ, Bire R, Hess P, Boer J (2009) Solid phase extraction for removal of matrix effects in lipophilic marine toxin analysis by liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem 394:1213–1226

    Article  CAS  Google Scholar 

  22. McCarron P, Kilcoyne J, Miles CO, Hess P (2009) Formation of azaspiracids-3, -4, -6, and -9 via decarboxylation of carboxyazaspiracid metabolites from shellfish. J Agric Food Chem 57:160–169

    Article  CAS  Google Scholar 

  23. Yasumoto T, Murata M, Oshima Y, Sano M, Matsumoto GK, Clardy J (1985) Diarrhetic shellfish toxins. Tetrahedron 41:1019–1025

    Article  CAS  Google Scholar 

  24. Mountfort DO, Suzuki T, Truman P (2001) Protein phosphatase inhibition adapted for determination of total DSP in contaminated mussel. Toxicon 39:383–390

    Article  CAS  Google Scholar 

  25. Doucet E, Ross N, Quilliam MA (2007) Enzymatic hydrolysis of esterified diarrhetic shellfish poisoning toxins and pectenotoxins. Anal Bioanal Chem 389:335–342

    Article  CAS  Google Scholar 

  26. Torgersen T, Wilkins AL, Rundberget T, Miles CO (2008) Characterization of fatty acid esters of okadaic acid and related toxins in blue mussels (Mytilus edulis) from Norway. Rapid Commun Mass Spectrom 22:1127–1136

    Article  CAS  Google Scholar 

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Acknowledgements

Technical assistance from Elliott Wright is gratefully acknowledged. Dr. Jeremy Melanson is thanked for reviewing this manuscript. This is NRCC manuscript number 51796.

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Authors and Affiliations

  1. National Research Council Canada, Institute for Marine Biosciences, 1411 Oxford Street, Halifax, NS, B3H 3Z1, Canada

    Pearse McCarron, Sabrina D. Giddings & Michael A. Quilliam

Authors
  1. Pearse McCarron
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  2. Sabrina D. Giddings
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  3. Michael A. Quilliam
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Correspondence to Pearse McCarron.

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McCarron, P., Giddings, S.D. & Quilliam, M.A. A mussel tissue certified reference material for multiple phycotoxins. Part 2: liquid chromatography–mass spectrometry, sample extraction and quantitation procedures. Anal Bioanal Chem 400, 835–846 (2011). https://doi.org/10.1007/s00216-011-4803-z

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  • DOI: https://doi.org/10.1007/s00216-011-4803-z

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