title: A mussel (Mytilus edulis) tissue certified reference material for the marine biotoxins azaspiracids.
abstract: Azaspiracids (AZAs) are lipophilic biotoxins produced by marine algae that can contaminate shellfish and cause human illness. The European Union (EU) regulates the level of AZAs in shellfish destined for the commercial market, with liquid chromatography-mass spectrometry (LC-MS) being used as the official reference method for regulatory analysis. Certified reference materials (CRMs) are essential tools for the development, validation, and quality control of LC-MS methods. This paper describes the work that went into the planning, preparation, characterization, and certification of CRM-AZA-Mus, a tissue matrix CRM, which was prepared as a wet homogenate from mussels (Mytilus edulis) naturally contaminated with AZAs. The homogeneity and stability of CRM-AZA-Mus were evaluated, and the CRM was found to be fit for purpose. Extraction and LC-MS/MS methods were developed to accurately certify the concentrations of AZA1 (1.16 mg/kg), AZA2 (0.27 mg/kg), and AZA3 (0.21 mg/kg) in the CRM. Quantitation methods based on standard addition and matrix-matched calibration were used to compensate for the matrix effects in LC-MS/MS. Other toxins present in this CRM at lower levels were also measured with information values reported for okadaic acid, dinophysistoxin-2, yessotoxin, and several spirolides.
authors.affiliation: National Research Council of Canada, Measurement Science and Standards, Biotoxin Metrology, 1411 Oxford Street, Halifax, NS, B3H 3Z1, Canada, pearse.mccarron@nrc-cnrc.gc.ca.
authors.forename: Pearse
authors.initials: P
authors.lastname: McCarron
authors.forename: Sabrina D
authors.initials: SD
authors.lastname: Giddings
authors.forename: Kelley L
authors.initials: KL
authors.lastname: Reeves
authors.forename: Philipp
authors.initials: P
authors.lastname: Hess
authors.forename: Michael A
authors.initials: MA
authors.lastname: Quilliam
mesh_terms: D000818:Animals; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods; D008387:Marine Toxins / Q000032:analysis* / Q000592:standards; D008978:Mollusk Venoms; D049880:Mytilus edulis / Q000737:chemistry*; D019319:Okadaic Acid / Q000032:analysis / Q000031:analogs & derivatives; D035964:Oxocins / Q000032:analysis; D011714:Pyrans / Q000032:analysis; D012015:Reference Standards; D013141:Spiro Compounds / Q000032:analysis* / Q000592:standards; D053719:Tandem Mass Spectrometry / Q000379:methods / Q000592:standards; D000095702:Polyether Toxins
chemical_list: D008387:Marine Toxins; D008978:Mollusk Venoms; D019319:Okadaic Acid; D035964:Oxocins; D011714:Pyrans; D013141:Spiro Compounds; C406592:azaspiracid; D000095702:Polyether Toxins; C066632:yessotoxin; C076221:dinophysistoxin 2