https://doi.org/10.4224/23002020
Vous avez des questions? Nous pouvons vous aider. Pour communiquer directement avec un auteur, consultez la
première page de la revue dans laquelle son article a été publié afin de trouver ses coordonnées. Si vous n’arrivez pas à les repérer, communiquez avec nous à PublicationsArchive-ArchivesPublications@nrc-cnrc.gc.ca.
Questions? Contact the NRC Publications Archive team at
PublicationsArchive-ArchivesPublications@nrc-cnrc.gc.ca. If you wish to email the authors directly, please see the first page of the publication for their contact information.
https://publications-cnrc.canada.ca/fra/droits
L’accès à ce site Web et l’utilisation de son contenu sont assujettis aux conditions présentées dans le site LISEZ CES CONDITIONS ATTENTIVEMENT AVANT D’UTILISER CE SITE WEB.
READ THESE TERMS AND CONDITIONS CAREFULLY BEFORE USING THIS WEBSITE. https://nrc-publications.canada.ca/eng/copyright
NRC Publications Archive Record / Notice des Archives des publications du CNRC :
https://nrc-publications.canada.ca/eng/view/object/?id=08994b47-6d85-4bd8-976c-a04ce72572e5 https://publications-cnrc.canada.ca/fra/voir/objet/?id=08994b47-6d85-4bd8-976c-a04ce72572e5
NRC Publications Archive
Archives des publications du CNRC
For the publisher’s version, please access the DOI link below./ Pour consulter la version de l’éditeur, utilisez le lien DOI ci-dessous.
Access and use of this website and the material on it are subject to the Terms and Conditions set forth at
Certified reference materials available from the NRCC Biotoxin Metrology program
Quilliam, Michael Arthur; McCarron, Pearse; Thomas, Krista; Crain, Sheila M.; Giddings, Sabrina; Beach, Daniel G.; LeBlanc, Pat; Calderon, Ruth Perez; Lewis, Nona Aisha; Reeves, Kelley; Hardstaff, William
Certified Reference Materials Available from the NRCC
Biotoxin Metrology Program
Measurement Science and Standards, Biotoxin Metrology
CRMs in italics are temporarily unavailable or in progress
Introduction
Ingestion of shellfish contaminated with toxins can result in diarrhea, amnesia, paralysis and even death. Routine monitoring prior to harvest is essential to ensure that seafood is safe for consumption and is necessary for international trade. Other algal toxins, originating from freshwater cyanobacteria, pose risks for drinking water. Testing for all these toxins requires validated methods, for which accurate calibration standards and certified reference materials (CRMs) are essential elements. The NRCC Biotoxin Metrology Program is a long running program that produces CRMs for marine and freshwater toxins under a rigorous quality system.
Marine toxins can be divided into two general classifications: hydrophilic and lipophilic. The hydrophilic toxins include the domoic acid and saxitoxin groups, which are responsible for amnesic shellfish poisoning (ASP) and paralytic shellfish poisoning (PSP), respectively. The lipophilic toxins include the okadaic acid group, pectenotoxins, azaspiracids, yessotoxins, and the cyclic imine group. The okadaic acid group of toxins is responsible for diarrhetic shellfish poisoning (DSP). Fresh and brackish water toxins comprise compounds from the microcystin, cylindrospermopsin, anatoxin, and saxitoxin groups.
This poster will provide an update on the biotoxin CRMs available from Biotoxin Metrology. There are two categories of materials: (a) calibration solution CRMs and (b) matrix CRMs. The former are critical for instrument calibration, while the latter are important for verifying the complete analytical method, from sample preparation and extraction through to data analysis.
Domoic Acid
Amnesic Shellfish Poisoning
Pseudonitzschia spp. H C H3 CH3 N H COOH COOH COOH Saxitoxin
Paralytic Shellfish Poisoning
Alexandrium tamarense O O N H2 N N N H2 + NH NH NH+2 OH OH H H Cl -Cl -Dinophysis spp. OkadaicAcid
Diarrhetic Shellfish Poisoning
O OH CH3 O OH OH O CH3 CH3 O O O H H OH OH CH3 O O CH3 Spirolide Alexandrium ostenfeldii Cyclic Imine Toxins
O O N O H O O O OH Pectenotoxin Dinophysis acuminata Pectenotoxins O O O O OH OH O O O O O OH O O Azaspiracid Azadinium spinosum Azaspiracid Shellfish Poisoning
O O O O O O O O O O O NaO3SO OH HO NaO3SO Protoceratium reticulatum Yessotoxins Yessotoxin Vulcanodinium rugosum Pinnatoxins Pinnatoxin-G Aphanizomenon issatschenkoi Anatoxin Anatoxin-a Microcystin-LR Microsystis aeruginosa Microcystins Cylindrospermopsis raciborskii Cylindrospermopsin Cylindrospermopsin NRC CRM-FDMT-1 homogeneity data.
Homogeneity and Stability
Homogeneity (ISO guide 30)
“A condition of being of uniform structure or composition with respect to one or more specified properties……”
Certification Methods
Values and uncertainties assigned following ISO guides
•
qNMR•
LC-CLND•
LC-MS/MS•
LC-UVD•
LC-pcox-FLD•
Accurate calibrationPreparati
on
of
Calibrati
on
Solutions
CR
Ms
Okadaic Acid/Dinophysistoxins (DSP toxins)
NRC CRM-OA okadaic acid
NRC CRM-DTX1 dinophysistoxin-1
NRC CRM-DTX2 dinophysistoxin-2
NRC CRM-DSP-Mus mussel tissue with DSP toxins
Pectenotoxins
NRC CRM-PTX2 pectenotoxin-2
Yessotoxins
NRC CRM-YTX yessotoxin
NRC CRM-hYTX 1-homo-yessotoxin
Cyclic Imine Toxins
NRC CRM-GYM gymnodimine NRC CRM-SPX1 13-desmethyl spirolide C NRC CRM-PnTX-G pinnatoxin-G
Azaspiracids
NRC CRM-AZA-1 azaspiracid-1 NRC CRM-AZA-2 azaspiracid-2 NRC CRM-AZA-3 azaspiracid-3NRC CRM-AZA-Mus mussel tissue with azaspiracids
Multi-toxin Freeze Dried Mussel Tissue
NRC CRM-FDMT-1
Paralytic Shellfish (PSP) Toxins
NRC CRM-C1&2 N-sulfocarbamoyl-gonyautoxin-1&2 NRC CRM-C3&4 N-sulfocarbamoyl-gonyautoxin-3&4 NRC CRM-dcGTX1&4 decarbamoylgonyautoxin-1&4 NRC CRM-dcGTX2&3 decarbamoylgonyautoxin-2&3 NRC CRM-dcNEO decarbamoylneosaxitoxin NRC CRM-dcSTX decarbamoylsaxitoxin NRC CRM-GTX1&4 decarbamoylgonyautoxin-1&4 NRC CRM-GTX2&3 decarbamoylgonyautoxin-2&3 NRC CRM-GTX5 gonyautoxin-5 NRC CRM-GTX6 gonyautoxin-6 NRC CRM-NEO neosaxitoxin NRC CRM-STX saxitoxin
NRC CRM-PSP-Mus mussel tissue with PSP toxins
Amnesic Shellfish (ASP) Toxins
NRC CRM-DA domoic acid
NRC CRM-iDAc iso-domoic acid C
NRC CRM-ASP-Mus mussel tissue with ASP toxins
Control Mussel Tissue
NRC CRM-Zero-Mus
Cylindrospermopsin
NRC CRM-CYN cylindrospermopsinMicrocystins
NRC CRM-MCRR microcystin-RR NRC CRM-MCLR microcystin-LR NRC CRM-dmMCLR [Dha7]-microcystin-LR NRC CRM-NODR nodularin-RAnatoxins
NRC CRM-ATX anatoxin-aNRC CRM-LWTX1 Lyngbya wollei saxitoxin analogue
NRC CRMs for Freshwater Biotoxins
NRC CRMs for Marine Biotoxins
Stability (ISO guide 30)
“Ability of a reference material, when stored under specified conditions, to maintain a stated property value within specified limits for a specified time……”
OA DTX1 DTX2 YTX AZA1 AZA2 AZA3 PTX2 SPX
13-desMe-C Average (mg/kg) 1.17 0.69 3.0 2.1 3.4 0.83 0.68 0.26 2.6 SD (n=20) 0.03 0.02 0.1 0.1 0.1 0.02 0.01 0.01 0.1 %RSD 2.8 2.9 2.3 4.7 1.6 1.8 1.7 5.6 3.6 µ*bb (%) 1.5 1.1 0.8 1.2 1.3 1.1 2.6 3.0 3.8
Preparati
on
of
Matri
x
CR
Ms
Acquisition of raw material (eg. shellfish, biomass, etc.)
Isolation and/or purity assessment
Quantitation of stock solution
Accurate dilution of stock solution
Dispensing in amber glass ampoules flushed with argon
Flame sealing of ampoules
Inspection and labelling of ampoules
Homogeneity and stability determination, characterization and certification.
Acquisition and profiling of raw material (eg. shellfish, biomass, etc.)
Combination of raw material and adjustment of moisture to 85%
Homogenisation
Degassing under N2
Dispensing aliquots while mixing & hermetic sealing of containers under N2
Capping bottles, inspection and labelling
Heat sealing bottles in foil pouches and transfer to proper storage
Homogeneity and stability determination, characterization and certification.
20 30 40 50 60 70 80 90 100 110 120 0 50 100 150 200 250 300 350 400 N o rmali ze d Days
Long Term Stability (OA in DSP-Mus-c) -80°C -2 stdev +2 stdev -20°C +4°C +20°c +40°C
Pearse McCarron, Krista Thomas, Sheila Crain, Sabrina Giddings, Daniel Beach, Pat LeBlanc,
Ruth Perez Calderon, Nancy Lewis, Kelley Reeves, William Hardstaff and Michael Quilliam
1411 Oxford Street, Halifax, Nova Scotia, B3H 3Z1, Canada
qNMR 1 2 3 4 4 ppm * * * * * 1 2 3 4 4 ppm 1 2 3 4 4 ppm 1 2 3 4 4 ppm * * * * * O O N H2 N H N N H2 + NH NH NH2+ OH OH H 1 3 7 9 10 12 6 Cl -Cl -Time (min) 0 2 4 6 8 10 12 14 STX NH4 + LC-CLND Time (min) 0 5 10 15 20 STX NEO dcSTX LC-pcox-FLD LC-MS/MS
OUR PARTNERS
Canadian Food Inspection AgencyBiotoxin Metrology’s quality system is recognized by SIM (Sistema Interamericano de Metrologia – Inter-american Metrology System), the Regional Metrology Organization for the Americas. SIM is committed to the \implementation of a Global Me\\asurement System within the Americas, in which all users can have confidence.
View publication stats View publication stats
