Determination of tetrodotoxin in puffer-fish tissues, and in serum and urine of intoxicated humans by liquid chromatography with tandem mass spectrometry

• Published in: Food Hygiene and Safety Science (Shokuhin Eiseigaku Zasshi) Vol. 47; no. 2; pp. 46 - 50
• Main Authors: Akaki, K.(Fukuoka-shi. Inst. for Hygiene and Environment (Japan)), Hatano, K
• Format: Journal Article
• Language: Japanese
• Published: Japan Japanese Society for Food Hygiene and Safety 01.04.2006
• Subjects: Adult; Aged; Aged, 80 and over; ANALYTICAL METHODS; Animals; BLOOD SERUM; Chromatography, Liquid; ESPECTROMETRIA DE MASAS; Female; Foodborne Diseases - blood; Foodborne Diseases - metabolism; Foodborne Diseases - urine; HPLC; Humans; LC/MS/MS; Male; MASS SPECTROMETRY; Middle Aged; ORINA; POISSON ARME; puffer-fish; PUFFERS (FISH); serum; SERUM SANGUIN; SPECTROMETRIE DE MASSE; SUERO SANGUINEO; TAMBOR (PEZ); TECHNIQUE ANALYTIQUE; TECNICAS ANALITICAS; Tetraodontiformes - metabolism; tetrodotoxin; Tetrodotoxin - analysis; Tetrodotoxin - blood; Tetrodotoxin - poisoning; Tetrodotoxin - urine; TOXINAS; TOXINE; TOXINS; URINE
• ISSN: 0015-6426; 1882-1006
• DOI: 10.3358/shokueishi.47.46

A simple and rapid method was developed for the analysis of tetrodotoxin in puffer-fish tissues, and in serum and urine of humans poisoned after consuming puffer-fish, by means of high-performance liquid chromatography with tandem mass spectrometry (LC/MS/MS). Tetrodotoxin was extracted with 2% acetic acid. The extracted solution from puffer-fish tissues was diluted with water, and the extracted solution from human serum and urine was cleaned up by LC/MS/MS with a methacrylate-styrenedivinylbenzene cartridge. The LC separation was performed on a C18 column (50 mm x 2.1 mm i.d.) using 10 mmol/L IPCC-MS7-methanol (65 : 35) as the mobile phase at a flow rate of 0.2 mL/min. The mass spectral acquisition was done in the positive ion mode by applying selected reaction monitoring (SRM). The recoveries of tetrodotoxin were 79-90% from puffer-fish tissues fortified at 0.1 mug/g and 1 mug/g, and 93-101% from human serum and urine fortified at 0.5 ng/mL and 5 ng/mL. The detection limits of tetrodotoxin were 0.01 mug/g in puffer-fish tissues and 0.1 ng/mL in human serum and urine. Thirty samples of puffer-fish from wholesale markets, and 7 serum and 5 urine samples of humans poisoned after consuming puffer-fish were analyzed by this method. Tetrodotoxin was detected in all puffer-fish tissues, and all serum and urine samples at the levels of 0.04-140 mug/g, 0.9-1.8 ng/mL and 15-150 ng/mL, respectively.