Simultaneous Determination of Tetrodotoxin in the Fresh and Heat-Processed Aquatic Products by High-Performance Liquid Chromatography–Tandem Mass Spectrometry

Abstract

Tetrodotoxin (TTX) was simultaneously detected in the fresh and heat-processed aquatic products by high-performance liquid chromatography–tandem mass spectrometry method. The detection conditions were investigated, including the chromatography column and mobile phase. Based on the optimized parameters, a sensitive determination method of TTX was established. The proposed method featured the merits of a good linear relationship between signal and TTX concentration (R2 = 0.9998), a wide detection matrix-based range of 0.2–100 ng/g, and a low detection limit of 0.2 ng/g, etc. The spiked assays evidenced its accuracy and reliability with recoveries of 90.5–107.2%. Finally, the developed method was simultaneously successfully applied in the determination of TTX in various fresh and heat-processed aquatic products.

Analytical Method Optimization of Tetrodotoxin and Its Contamination in Gastropods

“…The extraction of TTX from seafood mainly included two methods: boiling-assisted extraction with HCl or HAc/water [ 15 , 16 , 17 , 22 ] and ultrasound-assisted extraction with an HAc/methanol/water mixture [ 19 , 20 , 23 , 25 ]. The boiling-assisted extraction method has a good effect on protein precipitation, but there is a loss in the recovery of TTX.…”

“…Although the boiling time has decreased from 30 min [ 26 ] to 5 min [ 17 , 27 ], the method recoveries were only 70–84%. The recoveries of the ultrasound-assisted extraction method were 66.9–107% [ 19 , 20 , 23 ]. The recovery was related to the water content in the extract solvent mixture.…”

“…The immune affinity SPE cartridge can selectively adsorb TTX and effectively remove matrix interferences. It is suitable for sample pretreatment of low levels of TTX in seafood [ 19 , 20 ]. However, due to the limitation of column capacity, it is not suitable for the clean-up of high concentrations, especially toxic levels of TTX.…”

Tetrodotoxin and Its Analogues in Food: Recent Updates on Sample Preparation and Analytical Methods Since 2012

“…Analytical standards for TTX and its analogues are important because they are required for most analysis types. Certified reference materials for TTX are available from many sources: Funakoshi (purity = 96%, Tokyo, Japan), Nacalai tesque (purity ≥94%, Kyoto, Japan), Wako pure chemical industries (purity >99%, Osaka, Japan), Affixscientific Corporation (purity ≥98%, Fremont, CA, USA), Sigma-Aldrich (St. Louis, MO, USA), Cifga (Lugo, Spain), ,,, Tocris Bioscience (purity ≥98.8%, Bristol, UK), ,, Biorbyt Ltd. (Cambridge, UK), Abcam plc (Cambridge, UK), Latoxan (Valence, France), , J&K Scientific Ltd. (Beijing, China), Must Bio-Tech Co., Ltd. (Chengdu, China), , among others. Sometimes, TTX standards contain its analogues, for example, a TTX standard solution from CIFGA (Lugo, Spain) contains TTX, 4,9-anhydroTTX, 11-deoxyTTX, 4-epiTTX, and trace levels of 5,6,11-trideoxyTTX and 11-norTTX-6-ol .…”

“…Advantages and Limitations of Analytical Techniques of TTX and Its Analogues13,33,45,54,62,67,71,80,84 , high resolution, high sensitivity, excellent qualitative ability, high m/z analytes are extremely well resolved, unique capabilities of nontarget analysis and retrospectively review.TOF MS quantitative ability is inferior to QqQ MS, expensive instrumentation, complex maintenance, specialized personnel, TOF baselines are noisier than Orbitrap MS. not related to ion abundance, higher resolution than TOF MS, high sensitivity, excellent qualitative ability, low m/z analytes are extremely well resolved, multiplexing capability (Q-Orbitrap), unique capabilities of nontarget analysis and retrospectively review.Orbitrap MS quantitative ability is inferior to QqQ MS, expensive instrumentation, complex maintenance, specialized personnel, limited ion capacity, limited data acquisition speed when monitoring fast UHPLC peaks.…”