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Tetrodotoxin is a marine biotoxin with high acute toxicity. The levels in cooked seafood will help us to assess its intake in humans and may help assess the risk of toxicity. However, heavy matrices hinder the direct quantitation of tetrodotoxin. A quantitative method of measuring tetrodotoxin in cooked seafood using liquid chromatography with triple quadrupole mass spectrometry was established in this study. Tetrodotoxin was extracted from the sample matrix using 2% formic acid in methanol and cleaned using a cation exchange cartridge. The cleanup conditions were optimized. The matrix effects were determined using the postextraction spiking method and by comparing the slope of the linear regression equation in sample matrix to that in solvent. The limit of detection in the sample matrix was 5 μg/kg and the limit of quantification was 10 μg/kg. The mean recoveries at three spiking levels were 66.9-89.2% with relative standard deviations of 5.0-10.8% ( n = 6) in five different matrices. Tetrodotoxin was found at concentrations of 26.1-2462 μg/kg in nine of 83 cooked seafoods tested in this study. Eight analogs of Tetrodotoxin were detected in the samples studied. [ABSTRACT FROM AUTHOR]