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Screening for multiple classes of marine biotoxins by liquid chromatography–high-resolution mass spectrometry

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Abstract

Marine biotoxins pose a significant food safety risk when bioaccumulated in shellfish, and adequate testing for biotoxins in shellfish is required to ensure public safety and long-term viability of commercial shellfish markets. This report describes the use of a benchtop Orbitrap system for liquid chromatography–mass spectrometry (LC-MS) screening of multiple classes of biotoxins commonly found in shellfish. Lipophilic toxins such as dinophysistoxins, pectenotoxins, and azaspiracids were separated by reversed phase LC in less than 7 min prior to MS data acquisition at 2 Hz with alternating positive and negative scans. This approach resulted in mass accuracy for analytes detected in positive mode (gymnodimine, 13-desmethyl spirolide C, pectenotoxin-2, and azaspiracid-1, -2, and -3) of less than 1 ppm, while those analytes detected in negative mode (yessotoxin, okadaic acid, and dinophysistoxin-1 and -2) exhibited mass errors between 2 and 4 ppm. Hydrophilic toxins such as domoic acid, saxitoxin, and gonyautoxins were separated by hydrophilic interaction LC (HILIC) in less than 4 min, and MS data was collected at 1 Hz in positive mode, yielding mass accuracy of less than 1 ppm error at a resolving power of 100,000 for the analytes studied (m/z 300–500). Data were processed by extracting 5 ppm mass windows centered around the calculated masses of the analytes. Limits of detection (LOD) for the lipophilic toxins ranged from 0.041 to 0.10 μg/L (parts per billion) for the positive ions, 1.6–5.1 μg/L for those detected in negative mode, while the domoic acid and paralytic shellfish toxins yielded LODs ranging from 3.4 to 14 μg/L. Toxins were detected in mussel tissue extracts free of interference in all cases.

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Acknowledgements

The authors wish to thank Dr. Pearse McCarron for reviewing this manuscript and for providing extracts of the reference material containing lipophilic toxins. Loan of the Exactive™ mass spectrometer from Thermo Fisher Scientific is also gratefully acknowledged. This is NRC publication number 51791.

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Authors and Affiliations

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

    Pearl Blay, Joseph P. M. Hui & Jeremy E. Melanson

  2. Thermo Fisher Scientific, 355 River Oaks Parkway, San Jose, CA, 95134, USA

    James Chang

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  1. Pearl Blay
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  2. Joseph P. M. Hui
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  4. Jeremy E. Melanson
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Correspondence to Jeremy E. Melanson.

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Blay, P., Hui, J.P.M., Chang, J. et al. Screening for multiple classes of marine biotoxins by liquid chromatography–high-resolution mass spectrometry. Anal Bioanal Chem 400, 577–585 (2011). https://doi.org/10.1007/s00216-011-4772-2

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  • DOI: https://doi.org/10.1007/s00216-011-4772-2

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