Advertisement

Analytical and Bioanalytical Chemistry

, Volume 406, Issue 22, pp 5303–5312 | Cite as

Validation of a confirmatory method for lipophilic marine toxins in shellfish using UHPLC-HR-Orbitrap MS

  • Gabriel Orellana
  • Julie Vanden Bussche
  • Lieven Van Meulebroek
  • Michiel Vandegehuchte
  • Colin Janssen
  • Lynn VanhaeckeEmail author
Research Paper

Abstract

Lipophilic marine toxins are produced by harmful microalgae and can accumulate in edible filter feeders such as shellfish, leading to an introduction of toxins into the human food chain, causing different poisoning effects. During the last years, analytical methods, based on liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS), have been consolidated by interlaboratory validations. However, the main drawback of LC-MS/MS methods remains the limited number of compounds that can be analyzed in a single run. Due to the targeted nature of these methods, only known toxins, previously considered during method optimization, will be detected. Therefore in this study, a method based on ultra-high-performance liquid chromatography coupled to high-resolution Orbitrap mass spectrometry (UHPLC-HR-Orbitrap MS) was developed. Its quantitative performance was evaluated for confirmatory analysis of regulated lipophilic marine toxins in shellfish flesh according to Commission Decision 2002/657/EC. Okadaic acid (OA), dinophysistoxin-1 (DTX-1), pectenotoxin-2 (PTX-2), azaspiracid-1 (AZA-1), yessotoxin (YTX), and 13-desmethyl spirolide C (SPX-1) were quantified using matrix-matched calibration curves (MMS). For all compounds, the reproducibility ranged from 2.9 to 4.9 %, repeatability from 2.9 to 4.9 %, and recoveries from 82.9 to 113 % at the three different spiked levels. In addition, confirmatory identification of the compounds was effectively performed by the presence of a second diagnostic ion (13C). In conclusion, UHPLC-HR-Orbitrap MS permitted more accurate and faster detection of the target toxins than previously described LC-MS/MS methods. Furthermore, HRMS allows to retrospectively screen for many analogues and metabolites using its full-scan capabilities but also untargeted screening through the use of metabolomics software.

Keywords

Ultra-high-performance liquid chromatography high-resolution Orbitrap mass spectrometry Lipophilic marine toxins Okadaic acid Commission Decision 2002/657/EC Azaspiracids Yessotoxins 

Notes

Acknowledgments

The authors wish to thank Mieke Naessens, Ine Opsteyn, and Dirk Stockx for their technical contribution to this manuscript.

References

  1. 1.
    Paz B, Riobó P, Franco JM (2011) Rapid Commun Mass Spectrom 25:3627–3639CrossRefGoogle Scholar
  2. 2.
    Van Dolah FM (2000) Environ Health Perspect 108:133–141CrossRefGoogle Scholar
  3. 3.
    Scholin CA, Gulland F, Doucette GC, Benson S, Busman M, Chavez FP, Cordaro J, Delong R, De Vogelaere A, Harvey J, Haulena M, Lefebvre M, Lipscomb T, Loscutoff S, Lowenstine LJ, Marin R, Miller PE, McLellan WA, Moeller PDR, Powell CL, Rowles T, Silvagni P, Silver M, Spraker T, Trainer T, Van Dolah FM (2000) Nature 403:80–83CrossRefGoogle Scholar
  4. 4.
    Blay P, Hui JPM, Chang J, Melanson JE (2011) Anal Bioanal Chem 400:577–585CrossRefGoogle Scholar
  5. 5.
    Dominguez HJ, Paz B, Daranas AH, Norte M, Franco JM, Fernández LL (2010) Toxicon 56:191–217CrossRefGoogle Scholar
  6. 6.
    Garcia-Altares M, Diogene J, De la Iglesia P (2013) J Chromatogr A 1275:48–60CrossRefGoogle Scholar
  7. 7.
    Reguera B, Velo-Suárez L, Raine R, Park MG (2012) Harmful Algae 14:87–106CrossRefGoogle Scholar
  8. 8.
    Gerssen A, Mulder PPJ, de Boer J (2011) Anal Chim Acta 685:176–185CrossRefGoogle Scholar
  9. 9.
    Ciminiello P, Dell’Aversano C, Dello Iacovo E, Fattorusso E, Forino M, Tartaglione L, Rossi R, Soprano V, Capozzo D, Serpe L (2011) Anal Bioanal Chem 401:1043–1050CrossRefGoogle Scholar
  10. 10.
    Ciminiello P, Dell’Aversano C, Fattorusso E, Magno S, Tartaglione L, Cangini M, Pompei M, Guerrini F, Boni L, Pistocchi R (2006) Toxicon 47:597–604CrossRefGoogle Scholar
  11. 11.
    Jauffrais T, Kilcoyne J, Séchet V, Herrenknecht C, Truquet P, Hervé F, Baptiste Bérard J, Nulty C, Taylor S, Tillmann U, Milles CO, Hess P (2012) Mar Drugs 10:1382CrossRefGoogle Scholar
  12. 12.
    Suzuki T, Quilliam MA (2011) J Anal Sci 27:571–584CrossRefGoogle Scholar
  13. 13.
    EURLMB, EU-Harmonised SOP-LIPO-LC-MS/MS (2011) 4:1–31Google Scholar
  14. 14.
    Domènech A, Cortés-Francisco N, Palacios O, Franco JM, Riobó P, Llerena JJ, Vichi S, Caixach J (2014) J Chromatogr A 1328:16–25CrossRefGoogle Scholar
  15. 15.
    Škrabáková Z, O’Halloran J, van Pelt FNMA, James KJ (2010) Rapid Commun Mass Spectrom 24:2966–2974CrossRefGoogle Scholar
  16. 16.
    Vanhaecke L, Van Meulebroek L, De Clercq N, Vanden Bussche J (2013) Anal Chim Acta 767:118–127CrossRefGoogle Scholar
  17. 17.
    De Clerck N, Vanden Bussche J, Croubels S, Delahaut P, Vanhaecke L (2013) Chromatogr A 1301:111–121CrossRefGoogle Scholar
  18. 18.
    Wille K, De Brabander HF, De Wulff E, Van Caeter EP, Janssen CR, Vanhaecke L (2012) Trends Anal Chem 35:87–108CrossRefGoogle Scholar
  19. 19.
    Fux E, McMillan D, Bire R, Hess P (2007) J Chromatogr A 1157:273–280CrossRefGoogle Scholar
  20. 20.
    Fux E, Rode D, Bire R, Hess P (2008) Food Addit Contam Part A 25:1024–1032CrossRefGoogle Scholar
  21. 21.
    Commissions Decision 2002/657/EC (2002) Off J Eur Communities L221:8–36Google Scholar
  22. 22.
    Gerssen A, Van Olst EHW, Mulder PPJ, De Boer J (2010) Anal Bioanal Chem 397:3079–3088CrossRefGoogle Scholar
  23. 23.
    Bijttebier SKA, D’Hondt E, Hermans N, Apers S, Voorspoels S (2013) J Mass Spectrom 48:740–754CrossRefGoogle Scholar
  24. 24.
    Commission Decision 96/23/EC, Off (1996) J Eur Commun L125:10Google Scholar
  25. 25.
    Kilcoyne J, Fux E (2010) J Chromatogr A 1217:7123–7130CrossRefGoogle Scholar
  26. 26.
    Stobo LA, Lacaze JP, Scott AC, Gallacher S, Smith EA, Quilliam MA (2005) J AOAC Int 88:1371–1382Google Scholar
  27. 27.
    These A, Scholz J, Preiss-Weigert A (2009) J Chromatogr A 1216:4529–4538CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Gabriel Orellana
    • 1
    • 2
  • Julie Vanden Bussche
    • 1
  • Lieven Van Meulebroek
    • 1
  • Michiel Vandegehuchte
    • 2
  • Colin Janssen
    • 2
  • Lynn Vanhaecke
    • 1
    Email author
  1. 1.Faculty of Veterinary Medicine, Department of Veterinary Public Health and Food Safety, Laboratory of Chemical AnalysisGhent UniversityMerelbekeBelgium
  2. 2.Laboratory of Environmental Toxicology and Aquatic EcologyGhent UniversityGhentBelgium

Personalised recommendations