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Determination of tetrodotoxin by high performance liquid chromatography-tandem mass spectrometry

Objective To establish a new method for determination of tetrodotoxin(TTX) in puffer fish by high performance liquid chromatography- tandem mass spectrometry. Methods The sample was sonicated after homogenizing with 0.03 mol/L acetic acid solution. After a low-speed centrifugation, the supernatant was mixed with equivalent methanol. With a subsequent high-speed centrifugation, the supernatant was filtered through a 0.22 μm membrane filter before injection. LC-MS/MS analysis was conducted by Waters ACQUITY UPLC BEH Amide(2.1 mm× 100 mm, 1.7 μm) column with the mobile phase of 0.1% formic acid in water and acetonitrile at positive mode. Results There was a good linearity of TTX in the range of 5~1000 ng/m L. The matrix recoveries was 85.8%~93.2% and the absolute recoveries was 47.8%~48.9%. The detection limit is 5μg/kg and the limit of quantification is 2 0 μg/kg. Conclusion Compared with the previous methods, the new method is simple, rapid, accurate, sensitive and at low cost for the determination of TTX in aquatic products.