Determination of lipophilic marine toxins in mussels. Quantification and confirmation criteria using high resolution mass spectrometry.
A multitoxin method has been developed for quantification and confirmation of lipophilic marine biotoxins in mussels by liquid chromatography coupled to high resolution mass spectrometry (HRMS), using an Orbitrap-Exactive HCD mass spectrometer. Okadaic acid (OA), yessotoxin, azaspiracid-1, gymnodimine, 13-desmethyl spirolide C, pectenotoxin-2 and Brevetoxin B were analyzed as representative compounds of each lipophilic toxin group. HRMS identification and confirmation criteria were established. Fragment and isotope ions and ion ratios were studied and evaluated for confirmation purpose. In depth characterization of full scan and fragmentation spectrum of the main toxins were carried out. Accuracy (trueness and precision), linearity, calibration curve check, limit of quantification (LOQ) and specificity were the parameters established for the method validation. The validation was performed at 0.5 times the current European Union permitted levels. The method performed very well for the parameters investigated. The trueness, expressed as recovery, ranged from 80% to 94%, the precision, expressed as intralaboratory reproducibility, ranged from 5% to 22% and the LOQs range from 0.9 to 4.8pg on column. Uncertainty of the method was also estimated for OA, using a certified reference material. A top-down approach considering two main contributions: those arising from the trueness studies and those coming from the precision's determination, was used. An overall expanded uncertainty of 38% was obtained.
Food Science and Nutrition Department, XaRTA (Catalonian Reference Network on Food Technology), Campus de l'Alimentació de Torribera, University of Barcelona, Av/ Prat de la Riba, 171, 08921 Santa Coloma de Gramenet, Spain; Mass Spectrometry Laboratory/Organic Pollutants, IDAEA-CSIC, c/ JordiGirona, 18-26, 08034 Barcelona, Spain.
Albert
A
Domènech
Mass Spectrometry Laboratory/Organic Pollutants, IDAEA-CSIC, c/ JordiGirona, 18-26, 08034 Barcelona, Spain.
Nuria
N
Cortés-Francisco
Mass Spectrometry Laboratory/Organic Pollutants, IDAEA-CSIC, c/ JordiGirona, 18-26, 08034 Barcelona, Spain.
Oscar
O
Palacios
Instituto de Investigaciones Marinas [U. A. Fitoplancton Tóxico (CSIC-IEO)], Av/ E. Cabello, 6, 36208 Vigo, Pontevedra, Spain.
José M
JM
Franco
Instituto de Investigaciones Marinas [U. A. Fitoplancton Tóxico (CSIC-IEO)], Av/ E. Cabello, 6, 36208 Vigo, Pontevedra, Spain.
Pilar
P
Riobó
Quality Assurance Unit, CID-CSIC, c/ Jordi Girona, 18-26, 08034 Barcelona, Spain.
José J
JJ
Llerena
Food Science and Nutrition Department, XaRTA (Catalonian Reference Network on Food Technology), Campus de l'Alimentació de Torribera, University of Barcelona, Av/ Prat de la Riba, 171, 08921 Santa Coloma de Gramenet, Spain.
Stefania
S
Vichi
Mass Spectrometry Laboratory/Organic Pollutants, IDAEA-CSIC, c/ JordiGirona, 18-26, 08034 Barcelona, Spain. Electronic address: josep.caixach@cid.csic.es.
Josep
J
Caixach
D000818:Animals; D049872:Bivalvia / Q000737:chemistry*; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods; D008387:Marine Toxins / Q000032:analysis*; D013058:Mass Spectrometry / Q000379:methods; D049880:Mytilus edulis / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012758:Shellfish / Q000032:analysis
D008387:Marine Toxins
High resolution mass spectrometry; Identification and confirmation; Lipophilic marine toxins; Quantification; Uncertainty; Validation method