Determination of domoic acid in seawater and phytoplankton by liquid chromatography–tandem mass spectrometry

Abstract

Domoic acid (DA) is an algal neurotoxin produced by diatoms primarily of the genus Pseudo-nitzschia and is responsible for the human intoxication syndrome known as amnesic shellfish poisoning. A method has been developed to determine DA in seawater and phytoplankton matrices by liquid chromatography–tandem mass spectrometry for both quantitation and confirmation purposes. Sample extraction and clean-up was achieved on a C18 solid-phase extraction (SPE) cartridge. An acidic condition is critical for retaining hydrophilic DA on the cartridge. Direct injection of SPE eluate for analysis is recommended in order to avoid loss of DA by drying with heat prior to resuspension and injection. DA was quantified using the fragments produced from the protonated DA ion through multiple reaction monitoring (MRM). Recoveries exceeded 90% for all seawater samples spiked with DA and approximated 98% of toxin standard added to cultured phytoplankton material. Acceptable reproducibility (ca. 5% or less) was obtained for all intra-day and inter-day samples. The detection limit was 30 pg/ml level with a 20 μl injection volume, which demonstrated the value of this method for not only confirming DA production by minimally toxic phytoplankton species, but also for investigating the potentially important role of dissolved DA in marine food webs.