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 0.5g was treated with 5mL 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 (8.2min). 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.