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The preparation of certified calibration solutions for azaspiracid-1, -2, and -3, potent marine biotoxins found in shellfish

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Abstract

The production and certification of a series of azaspiracid (AZA) calibration solution reference materials is described. Azaspiracids were isolated from contaminated mussels, purified by preparative liquid chromatography and dried under vacuum to the anhydrous form. The purity was assessed by liquid chromatography–mass spectrometry and nuclear magnetic resonance spectroscopy. The final concentration of each AZA in a CD3OH stock solution was determined by quantitative NMR spectroscopy. This solution was then diluted very accurately in degassed, high purity methanol to a concentration of 1.47 ± 0.08 μmol/L for CRM-AZA1, 1.52 ± 0.05 μmol/L for CRM-AZA2, and 1.37 ± 0.13 μmol/L for CRM-AZA3. Aliquots were dispensed into argon-filled glass ampoules, which were immediately flame-sealed. The calibration solutions are suitable for method development, method validation, calibration of liquid chromatography or mass spectrometry instrumentation and quality control of shellfish monitoring programs.

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Acknowledgments

The first two authors contributed equally to this work. The authors would like to thank all members of the NRC Certified Reference Materials Program for assistance with ampouling the AZA calibration solutions. Funding for materials retrieval and isolation of AZAs was provided by the Irish National Development Plan through the ASTOX project (ST/02/02, 2003–2007), and the ASTOX2 project (Irish National Development Plan 2007–2013, grant number PBA/AF/08/001(01)). This is NRC publication number 51766.

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Author notes

  1. Nils Rehmann

    Present address: Research and Productivity Council of New Brunswick, 921 College Hill Rd., Fredericton, NB, E3B 6Z9, Canada

  2. Philipp Hess

    Present address: Ifremer, Centre de Nantes, Département Environnement Microbiologie et Phycotoxines, Rue de l’Ile d’Yeu, 44311, Nantes, Cedex 03, France

Authors and Affiliations

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

    Ruth A. Perez, Sheila Crain, Patricia LeBlanc, Cheryl Craft, Shawna MacKinnon, Kelley Reeves, Ian W. Burton, John A. Walter, Michael A. Quilliam & Jeremy E. Melanson

  2. Marine Institute, Rinville, Oranmore, Co., Galway, Ireland

    Nils Rehmann & Philipp Hess

Authors
  1. Ruth A. Perez
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  2. Nils Rehmann
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  3. Sheila Crain
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  4. Patricia LeBlanc
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  5. Cheryl Craft
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  6. Shawna MacKinnon
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  7. Kelley Reeves
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  8. Ian W. Burton
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  9. John A. Walter
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  10. Philipp Hess
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  11. Michael A. Quilliam
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Corresponding author

Correspondence to Jeremy E. Melanson.

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Perez, R.A., Rehmann, N., Crain, S. et al. The preparation of certified calibration solutions for azaspiracid-1, -2, and -3, potent marine biotoxins found in shellfish. Anal Bioanal Chem 398, 2243–2252 (2010). https://doi.org/10.1007/s00216-010-4161-2

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  • DOI: https://doi.org/10.1007/s00216-010-4161-2

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