Development of a method for the identification of azaspiracid in shellfishby liquid chromatography-tandem mass spectrometry

Azaspiracid is the main toxin responsible for a number of recent human into xications in Europe resulting from shellfish consumption. The first micro l iquid chromatography-tandem mass spectrometry (micro-LC-MS-MS) method was d eveloped for the determination of this novel shellfish poisoning toxin in m ussels. The analyte was extracted from whole mussel meat with acetone and c hromatographed on a C-18 reversed-phase column (1.0 mm I.D.) by isocratic e lution at 30 mu l/min with acetonitrile-water (85:15, v/v), containing 0.03 % trifluoroacetic acid. The toxin was ionised in an ionspray interface oper ating in the positive ion mode, where only the intact protonated molecule, [M+H](+), was generated at m/z 842. This served as precursor ion for collis ion-induced dissociation and three product ions, [M+H-nH(2)O](+) with n=1-3 , were identified for the unambiguous toxin confirmation by selected reacti on monitoring LC-MS-MS analysis. A detection limit of 20 pg, based on a 3:1 signal-to-noise ratio, was achieved for the analyte. This LC-MS-MS method was successfully applied to determine azaspiracid in toxic cultivated shell fish from two regions of Ireland. (C) 2000 Elsevier Science B.V. All rights reserved.