title: Determination of azaspiracids in shellfish using liquid chromatography/tandem electrospray mass spectrometry.
abstract: Azaspiracid (AZA1), a recently discovered marine toxin, is responsible for the new human toxic syndrome, azaspiracid poisoning (AZP), which is caused by the consumption of contaminated shellfish. A new, sensitive liquid chromatography/mass spectrometry (LC/MS) method has been developed for the determination of AZA1 and its analogues, 8-methylazaspiracid (AZA2) and 22-demethylazaspiracid (AZA3). Separation of these toxins was achieved using reversed-phase LC and coupled, via an electrospray ionisation (ESI) source, to an ion-trap mass spectrometer. Spectra showed the protonated molecules, [M + H]+, and their major product ions, due to the sequential loss of two water molecules, [M + H - H2O]+, [M + H - 2H2O]+, in addition to fragment ions that are characteristic of these cyclic polyethers. A highly specific and sensitive LC/MS(3) analytical method was developed and, using shellfish extracts containing AZA1, the detection limit (S/N = 3) was 4 pg on-column, corresponding to 0.8 ng/mL. Using the protocol presented here, this is equivalent to 0.37 ng/g shellfish tissue and good linear calibrations were obtained for AZA1 in shellfish extracts (average r2 = 0.9988). Good reproducibility was achieved with % RSD values (N = 5) ranging from 1.5% (0.75 microg/mL) to 4.2% (0.05 microg/mL). An efficient procedure for the extraction of toxins from shellfish aided the development of a rapid protocol for the determination of the three predominant azaspiracids.
authors.affiliation: Ecotoxicology Research Unit, Chemistry Department, Cork Institute of Technology, Cork, Ireland.
authors.forename: Ambrose
authors.initials: A
authors.lastname: Furey
authors.forename: Ana
authors.initials: A
authors.lastname: Braña-Magdalena
authors.forename: Mary
authors.initials: M
authors.lastname: Lehane
authors.forename: Cian
authors.initials: C
authors.lastname: Moroney
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: James
authors.forename: Masayuki
authors.initials: M
authors.lastname: Satake
authors.forename: Takeshi
authors.initials: T
authors.lastname: Yasumoto
mesh_terms: D000818:Animals; D002138:Calibration; D002853:Chromatography, Liquid; D004064:Digestive System / Q000737:chemistry; D005517:Foodborne Diseases; D007202:Indicators and Reagents; D008387:Marine Toxins / Q000032:analysis*; D008460:Meat / Q000032:analysis; D012758:Shellfish / Q000032:analysis*; D021241:Spectrometry, Mass, Electrospray Ionization; D013141:Spiro Compounds / Q000032:analysis*; D000095702:Polyether Toxins
chemical_list: D007202:Indicators and Reagents; D008387:Marine Toxins; D013141:Spiro Compounds; C406592:azaspiracid; D000095702:Polyether Toxins