Simultaneous determination of biotoxins DSP and AZAs in bivalve molluscs and fish by liquid chromatography/tandem mass spectrometry.

Abstract

A method has been developed for simultaneous determination of the toxins OA, DTX-1, AZA-1, AZA-2 and AZA-3 in various aquatic products as these can cause diarrhoetic shellfish poisoning (DSP) in humans, an intoxication characterized by vomiting and diarrhea. Separation of the toxins was achieved on a C18 column (150 mm × 2.1 mm, 3.5 µm) using an acetonitrile/water gradient with formic acid as an eluent modifier. Electrospray ionisation (ESI) in negative mode was used to generate the molecule related ion [M-H](-), for OA and DTX-1, while ESI in positive mode was used to generate the molecule related ion [M+H](+) for AZAs. Samples were extracted with 80% methanol, followed by partitioning with ethyl acetate, purified on a Poly-Sery MAX cartridge and finally analyzed by LC/ESI-MS/MS in the multiple reaction monitoring (MRM) mode. The limit of detection (LOD) and limit of qualification (LOQ) of the method were in the range of 0.02-0.79 µg/kg and 0.07-2.64 µg/kg in Scomberomorus niphonius, blood clam and oyster, respectively, recoveries of the toxins at three fortification levels ranged from 71.3% to 104.8% with relative standard deviation from 1.0% to 12.5%. The calibration curves were well linear between the LC peak area of the selected ion pair and the concentration of the toxins, with the correlation coefficient over 0.99. The method was sufficiently sensitive to permit the determination of the toxins DSP and AZA in sea food.