First determination of extracellular paralytic shellfish poisoning toxins in the culture medium of toxigenic dinoflagellates by HILIC-HRMS.

Paralytic shellfish poisoning (PSP) toxins have received considerable attention in recent years because of their adverse effects on marine breeding industries and human health. In this study, a reliable method for the analysis of extracellular PSP toxins in the culture medium of marine toxic dinoflagellates was developed for the first time using graphitized carbon black-solid-phase extraction and hydrophilic interaction liquid chromatography-high-resolution mass spectrometry. The limit of quantification of typical PSP toxins in algal culture medium ranged from 0.072 μg/L to 0.151 μg/L under optimal conditions. Satisfactory absolute recoveries (87.5%-102.4%), precision (relative standard deviation ≤ 7.6%), and linearity (R ≥ 0.9998) were also achieved. In addition, the proposed method was applied to screen and determine the extracellular PSP toxins of two typical toxigenic dinoflagellates, Alexandrium minutum and Alexandrium tamarense. The total concentrations of the extracellular PSP toxins in A. minutum and A. tamarense over the whole growth period were within 2.0-735.5 and 2.0-19.2 μg/L, respectively. The concentrations of extracellular PSP toxins varied remarkably in the different growth stages of A. minutum and A. tamarense, and the contents of some extracellular PSP toxins were substantially higher than those of intracellular PSP toxins. Therefore, the extracellular PSP toxins released by toxigenic red tide algae cannot be ignored, and their environmental fate, bioavailability, and potential harm to aquatic environment need to be investigated in future studies.