title: Liquid chromatography--multiple tandem mass spectrometry for the determination of ten azaspiracids, including hydroxyl analogues in shellfish.
abstract: Azaspiracids (AZAs) are a group of polyether toxins that cause food poisoning in humans. These toxins, produced by marine dinoflagellates, accumulate in filter-feeding shellfish, especially mussels. Sensitive liquid chromatography-electrospray ionisation mass spectrometry (LC-ESI-MS(n)) methods have been developed for the determination of the major AZAs and their hydroxyl analogues. These methods, utilising both chromatographic and mass resolution, were applied for the determination of 10 AZAs in mussels (Mytilus edulis). An optimised isocratic reversed phase method (3 microm Luna-2 C18 column) separated 10 azaspiracids using acetonitrile/water (46:54, v/v) containing 0.05% trifluoroacetic acid (TFA) and 0.004% ammonium acetate in 55 min. Analyte determination using MS3 involved trapping and fragmentation of the [M + H]+ and [M + H - H2O]+ ions with detection of the [M + H - 2H2O]+ ion for each AZA. Linear calibrations were obtained for AZA1, using spiked shellfish extracts, in the range 0.05-1.00 microg/ml (r2 = 0.997) with a detection limit of 5 pg (signal : noise = 3). The major fragmentation pathways in hydroxylated azaspiracids were elucidated using hydrogen/deuterium (H/D) exchange experiments. An LC-MS3 method was developed using unique parent ions and product ions, [M + H - H2O - CgH10O2R1R3]+, that involved fragmentation of the A-ring. This facilitated the discrimination between 10 azapiracids, AZA1-10. Thus, this rapid LC-MS3 method did not require complete chromatographic resolution and the run-time of 7 min had detection limits better than 20 pg for each toxin.
authors.affiliation: PROTEOBIO, Mass Spectrometry Centre for Proteomics and Biotoxin Research, Department of Chemistry, Cork Institute of Technology, Bishopstown, Cork, Ireland.
authors.forename: Mary
authors.initials: M
authors.lastname: Lehane
authors.forename: María José
authors.initials: MJ
authors.lastname: Fidalgo Sáez
authors.forename: Ana Braña
authors.initials: AB
authors.lastname: Magdalena
authors.forename: Isabel
authors.initials: I
authors.lastname: Ruppén Cañás
authors.forename: Mónica
authors.initials: M
authors.lastname: Díaz Sierra
authors.forename: Brett
authors.initials: B
authors.lastname: Hamilton
authors.forename: Ambrose
authors.initials: A
authors.lastname: Furey
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: James
mesh_terms: D000818:Animals; D049872:Bivalvia / Q000737:chemistry*; D002138:Calibration; D002850:Chromatography, Gel / Q000379:methods*; D008387:Marine Toxins / Q000032:analysis* / Q000145:classification; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013141:Spiro Compounds / Q000032:analysis* / Q000145:classification; D000095702:Polyether Toxins
chemical_list: D008387:Marine Toxins; D013141:Spiro Compounds; C406592:azaspiracid; D000095702:Polyether Toxins