Screening of tetrodotoxin in puffers using gas chromatography–mass spectrometry

Tetrodotoxin (TTX), a toxic compound found in some puffers can cause death to humans through consumption. We have developed a simplified method for the screening of TTX in puffers using GC–MS. A puffer tissue of 0.5 g was treated with 5 mL of 0.1% acetic acid, followed by alkaline hydrolysis, LLE or liquid–liquid extraction and N-methyl-N-TMS-trifluoroacetamide derivatization. The developed method used only a small sample and solvent, simplified LLE and derivatization procedures and short chromatographic analysis (8.2 min). All of these contribute to cost-saving, enhanced sample throughput and high sensitivity of the screening assay. The developed method was validated and proved to be within the acceptable range.