Temperature Dependence of the Pre-Chromatographic ‘Lawrence’ Method for Bivalves Contaminated with Paralytic Shellfish Poisoning Toxins

Saxitoxins are potent neurotoxins originating the acute human neurological syndrome of paralytic shellfish poisoning (PSP) via bivalve vectors. The official testing method in the European Union, commonly known as the ‘Lawrence method’, involves pre-column oxidation steps. The Portuguese monitoring adopted the hydrogen peroxide oxidation screening approach for bivalves contaminated with Gymnodinium catenatum toxins, which can quantify chromatographically at once 6 out of 10 analogues commonly found in bivalves. Seasonal fluctuation in the fluorescence yield of calibration curves was observed across years in a consistent manner. It correlated with fluctuations in average monthly air temperature in Lisbon, highlighting the importance of recording the room temperature during the oxidation steps as a matter of routine practice. Incubation experiments also showed an increase in fluorescence yield with temperature, more pronounced for the 11-hydroxysulphate analogues (dcGTX2 + 3, C1 + 2, GTX2 + 3) than for the 11-H toxins (dcSTX, GTX5[B1], STX). Temperature can be exploited to increase fluorescence yield, assisting in spectral confirmation, but must not exceed 40–50 °C to avoid toxin decomposition or production of extra oxidation products.