Skip to main content

Advertisement

SpringerLink
Log in

Development and single laboratory validation of an optical biosensor assay for tetrodotoxin detection as a tool to combat emerging risks in European seafood

  • Research Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

Tetrodotoxin (TTX) is a potent neurotoxin emerging in European waters due to increasing ocean temperatures. Its detection in seafood is currently performed as a consequence of using the Association of Analytical Communities (AOAC) mouse bioassay (MBA) for paralytic shellfish poisoning (PSP) toxins, but TTX is not monitored routinely in Europe. Due to ethical and performance-related issues associated with this bioassay, the European Commission has recently published directives extending procedures that may be used for official PSP control. An AOAC-accredited high-performance liquid chromatography (HPLC) method has now been accepted by the European Union as a first action screening method for PSP toxins to replace the MBA. However, this AOAC HPLC method is not capable of detecting TTX, so this potent toxin would be undetected; thereby, a separate method of analysis is required. Surface plasmon resonance (SPR) optical biosensor technology has been proven as a potential alternative screening method to detect PSP toxins in seafood. The addition of a similar SPR inhibition assay for TTX would complement the PSP assay in removing the MBA. The present report describes the development and single laboratory validation in accordance with AOAC and IUPAC guidelines of an SPR method to be used as a rapid screening tool to detect TTX in the sea snail Charonia lampas lampas, a species which has been implicated in 2008 in the first case of human TTX poisoning in Europe. As no current regulatory limits are set for TTX in Europe, single laboratory validation was undertaken using those for PSP toxins at 800 μg/kg. The decision limit (CCα) was 100 μg/kg, with the detection capability (CCβ) found to be ≤200 μg/kg. Repeatability and reproducibility were assessed at 200, 400, and 800 μg/kg and showed relative standard deviations of 8.3, 3.8, and 5.4 % and 7.8, 8.3, and 3.7 % for both parameters at each level, respectively. At these three respective levels, the recovery of the assay was 112, 98, and 99 %.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Moczydlowski EG (2013) The molecular mystique of TTX. Toxicon 63:165–183

    Article  CAS  Google Scholar 

  2. Yasumoto T, Yotsu-Yamashita M (1996) Chemical and etiological studies on TTX and its analogs. Toxin Rev 15(2):81–90

    Article  CAS  Google Scholar 

  3. Fozzard HA, Lipkind GM (2010) The TTX binding site is within the outer vestibule of the sodium channel. Mar Drugs 8:219–234

    Article  CAS  Google Scholar 

  4. Williams BL (2010) Behavioral and chemical ecology of marine organisms with respect to tetrodotoxin. Mar Drugs 8:381–398

    Article  CAS  Google Scholar 

  5. French RJ, Yoshikami D, Sheets MF, Olivera BM (2010) The tetrodotoxin receptor of voltage-gated sodium channels—perspectives from interactions with μ-conotoxins. Mar Drugs 8:2153–2161

    Google Scholar 

  6. Lee HL, Ruben PC (2008) Interaction between voltage-gated sodium channels and the neurotoxin, TTX. Channels 2:407–412

    Article  Google Scholar 

  7. Narahashi T (2001) Pharmacology of TTX. Toxin Rev 20:67–84

    Article  CAS  Google Scholar 

  8. Narahashi T (2008) TTX: a brief history. Proc Jpn Acad Ser B Phys Biol Sci 84:147–154

  9. Noguchi T, Arakawa O (2008) TTX—distribution and accumulation in aquatic organisms, and cases of human intoxication. Mar Drugs 6:220–242

    Article  CAS  Google Scholar 

  10. Saoudi M, Abdelmouleh A, El Feki A (2010) TTX: a potent marine toxin. Toxin Rev 29(2):60–70

    Article  CAS  Google Scholar 

  11. Alcaraz A, Whipple RE, Gregg HR, Andresen BD, Grant PM (1999) Analysis of TTX. Forensic Sci Int 99(1):35–45

    Article  CAS  Google Scholar 

  12. How CK, Chern CH, Huang YC, Wang LM, Lee CH (2003) TTX poisoning. Am J emerg med 21:51–54

    Article  Google Scholar 

  13. Hungerford JM, Committee on Natural Toxins and Food Allergens (2006) Marine and freshwater toxins. J AOAC Int 89(1):248–269

    CAS  Google Scholar 

  14. Yakes BJ, Deeds J, White K, Degrasse SL (2011) Evaluation of surface plasmon resonance biosensors for detection of TTX in food matrices and comparison to analytical methods. J Agric Food Chem 59(3):839–846

    Article  CAS  Google Scholar 

  15. Rodriguez P, Alfonso A, Vale C, Alfonso C, Vale P, Tellez A, Botana LM (2008) First toxicity report of TTX and 5,6,11-trideoxyTTX in the trumpet shell Charonia lampas lampas in Europe. Anal Chem 80:5622–5629

    Article  CAS  Google Scholar 

  16. Fernández-Ortega JF, Morales-de los Santos JM, Herrera-Gutiérrez ME, Fernández-Sánchez V, Rodríguez Loureo P, Alfonso Rancaño A, Téllez-Andrade A (2010) Seafood intoxication by TTX: first case in Europe. J Emerg Med 39:612–617

    Article  Google Scholar 

  17. Bentur Y, Ashkar J, Lurie Y, Levy Y, Azzam ZS, Litmanovich M, Golik M, Gurevych B, Golani D, Eisenman A (2008) Lessepsian migration and TTX poisoning due to Lagocephalus sceleratus in the eastern Mediterranean. Toxicon 52:964–968

    Article  CAS  Google Scholar 

  18. Katikou P, Georgantelis D, Sinouris N, Petsi A, Fotaras T (2009) First report on toxicity assessment of the Lessepsian migrant pufferfish Lagocephalus sceleratus (Gmelin, 1789) from European waters (Aegean Sea, Greece). Toxicon 54:50–55

    Article  CAS  Google Scholar 

  19. Rodriguez P, Alfonso A, Otero P, Katikou P, Georgantelis D, Botana LM (2012) Liquid chromatography–mass spectrometry method to detect TTX and its analogues in the puffer fish Lagocephalus sceleratus (Gmelin, 1789) from European waters. Food Chem 132(2):1103–1111

    Article  CAS  Google Scholar 

  20. Silva M, Azevedo J, Rodriguez P, Alfonso A, Botana LM, Vasconcelos V (2012) New gastropod vectors and TTX potential expansion in temperate waters of the Atlantic Ocean. Mar drugs 10:712–726

    Article  CAS  Google Scholar 

  21. Commission Regulation (EC) no. 854/2004 of the European Parliament and of the Council of 29 April (2004) Laying down specific rules for the organisation of official controls on products of animal origin intended for human consumption. Off J Eur Communities L226:83–126

    Google Scholar 

  22. AOAC International (2005) AOAC official method 959.08. In: Official methods of analysis of AOAC International, section 49.10.01, 18th ed. AOAC International, Gaithersburg

  23. Yasumoto T (1991) The manual for methods of food sanitation tests. Japan Food Hygienic Association, Tokyo

    Google Scholar 

  24. AOAC International (2005) AOAC official method 2005.06. In: Official methods of analysis of AOAC International, section 49.10.03, 18th ed. AOAC International, Gaithersburg

  25. Doucette GJ, Powell CL, Do EU, Byon CY, Cleves F, McClain SG (2000) Evaluation of 11-[H3]-TTX use in a heterologous receptor binding assay for PSP toxins. Toxicon 38:1465–1474

    Article  CAS  Google Scholar 

  26. Chen CY, Chou HN (1998) Detection of TTX by high performance liquid chromatography in lined moon shell and puffer fish. Acta Zool Taiwanica 9(1):41–48

    CAS  Google Scholar 

  27. Kouassi Nzoughet J, Campbell K, Barnes P, Cooper KM, Chevallier OP, Elliott CT (2013) Comparison of sample preparation methods and validation of an UPLC-MS/MS procedure to EU guidelines for the quantification of TTX present in marine gastropods and pufferfish. Food Chem 136:1584–1589

    Article  Google Scholar 

  28. Kawatsu K, Hamano Y, Yoda T, Teano Y, Shibata T (1997) Rapid and highly sensitive enzyme immunoassay for quantitative determination of TTX. Jpn J Med Sci Biol 50:133–150

    CAS  Google Scholar 

  29. Matsumura K, Fukiya S (1992) Indirect competitive enzyme immunoassay for TTX using a biotin avidin system. J AOAC Int 75(5):883–886

    CAS  Google Scholar 

  30. Raybould TJG, Bignami GS, Inouye LK, Simpson SB, Byrnes JB, Grothaus PG, Vann DC (1992) A monoclonal antibody based immunoassay for detecting TTX in biological samples. J Clin Lab Anal 6:65–72

    Article  CAS  Google Scholar 

  31. Tao J, Wei WJ, Nan L, Lei LH, Hui HC, Fen GX, Jun LY, Jing Z, Rong J (2010) Development of competitive indirect ELISA for the detection of TTX and a survey of the distribution of TTX in the tissues of wild puffer fish in the waters of south-east China. Food Addit Contam 27(11):1589–1597

    Article  CAS  Google Scholar 

  32. Taylor AD, Ladd J, Etheridge S, Deeds J, Hall S, Jiang S (2008) Quantitative detection of TTX (TTX) by a surface plasmon resonance (SPR) sensor. Sensors Actuators B 130:120–128

    Article  CAS  Google Scholar 

  33. Vaisocherova H, Taylor AD, Jiang S, Hegnerova K, Vala M, Homola J, Yakes BJ, Deeds J, DeGrasse S (2011) Surface plasmon resonance biosensor for determination of TTX: prevalidation study. J AOAC Int 94:596–604

    Google Scholar 

  34. Traynor I, Plumpton L, Fodey TL, Higgins C, Elliott CT (2006) Immunobiosensor detection of domoic acid as a screening test in bivalve mollusks: comparison with LC-based analysis. J AOAC Int 89:868–872

    CAS  Google Scholar 

  35. Stewart LD, Hess P, Connolly L, Elliott CT (2009) Development and single-laboratory validation of a pseudofunctional biosensor immunoassay for the detection of the okadaic acid group of toxins. Anal Chem 81:10208–10214

    Article  CAS  Google Scholar 

  36. Campbell K, Haughey SA, van den Top H, van Egmond H, Vilarino N, Botana LM, Elliott CT (2010) Single laboratory validation of a surface plasmon resonance biosensor screening method for paralytic shellfish poisoning toxins. Anal Chem 82:2977–2988

    Article  CAS  Google Scholar 

  37. Van den Top H, Haughey S, Vilariño N, Botana L, Van Egmond H, Elliott CT, Campbell K (2011) Surface plasmon resonance biosensor screening method for paralytic shellfish poisoning toxins: a pilot interlaboratory study. Anal Chem 83:4206–4213

    Article  Google Scholar 

  38. Campbell K, Stewart LD, Fodey TL, Haughey SA, Doucette GJ, Kawatsu K, Elliott CT (2007) An assessment of specific binding proteins suitable for the detection of paralytic shellfish poisons (PSP) using optical biosensor technology. Anal Chem 79(15):5906–5914

    Article  CAS  Google Scholar 

  39. Bates HA, Kostriken R, Rapoport H (1978) A chemical assay for saxitoxin. Improvements and modifications. J Agric food chem 26:252–254

    Article  CAS  Google Scholar 

  40. Thompson M, Ellison SLR, Wood R (2002) Harmonized guidelines for single-laboratory validation of methods of analysis (IUPAC technical report). Pure Applied Chemistry 74:835–855

    Article  CAS  Google Scholar 

  41. AOAC guidelines for single laboratory validation of chemical methods for dietary supplements and botanicals. Available at http://www.aoac.org/Official_Methods/slv_guidelines.pdf. Accessed 22 April 2013

  42. European Commission Decision (EC) no. 2002/657/EC implementing Council Directive 96/23/EC concerning the performance of analytical methods and the interpretation of results. Official Journal of the European Communities L221:8–34

Download references

Acknowledgments

This research was partly funded through The Interreg Project “ATLANTOX: Advanced Tests about New Toxins appeared in the Atlantic Area” and the CONffIDENCE: EU FP7 project “CONtaminants in Food and Feed: Inexpensive Detection for Control of Exposure” financially supported by the European Commission (grant agreement number 211326—collaborative project). The authors would like to thank Panagiota Katikou of the National Reference Laboratory for Marine Biotoxins in Greece for the provision of puffer fish samples.

Author information

Authors and Affiliations

  1. Institute for Global Food Security, School of Biological Sciences, Queen’s University, David Keir Building, Stranmillis Road, Belfast, BT9 5AG, UK

    Katrina Campbell, Simon A. Haughey & Christopher T. Elliott

  2. UK National Reference Laboratory for Marine Biotoxins, Agri-Food and Biosciences Institute, Stoney Road, Stormont, BT4 3SD, UK

    Paul Barnes & Cowan Higgins

  3. Division of Bacteriology, Osaka Prefectural Institute of Public Health, 3-69, Nakamichi 1-chome, Higashinari-ku, Osaka, 537-0025, Japan

    Kentaro Kawatsu

  4. Centro Interdisciplinar de Investigacao Marinha e Ambiental (CIIMAR), Rua dos Bragas, no. 289, 4050-123, Porto, Portugal

    Vitor Vasconcelos

Authors
  1. Katrina Campbell
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Paul Barnes
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Simon A. Haughey
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Cowan Higgins
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kentaro Kawatsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Vitor Vasconcelos
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Christopher T. Elliott
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Katrina Campbell.

Additional information

Published in the topical collection Rapid Detection in Food and Feed with guest editors Rudolf Krska and Michel Nielen.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Campbell, K., Barnes, P., Haughey, S.A. et al. Development and single laboratory validation of an optical biosensor assay for tetrodotoxin detection as a tool to combat emerging risks in European seafood. Anal Bioanal Chem 405, 7753–7763 (2013). https://doi.org/10.1007/s00216-013-7106-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-013-7106-8

Keywords

Search

Navigation