Determination of Tetrodotoxin in Fugu Liver Using Hydrophilic Interaction Liquid Chromatography Coupled with Triple Quadrupole Mass Spectrometry

Abstract:

Objective: To develop a LC-MS-MS method for determination of tetrodotoxin (TTX) in Fugu liver. Method: The analyte was vortex-mixed, extracted with acetonitrile by ultrasound, and followed by centrifugation. The supernatant (5 μL) was injected into the LC-MS-MS system for analysis. The separation was performed on an Innovation HP Amide column (100 mm×3.0 mm, 5 μm) at 30 ℃ using acetonitrile-0.3% formic acid (70:30) as mobile phase at a flow rate of 0.5 mL·min^-1. Quantification was through tandem-mass spectrometry with positive electro-spray ionization (ESI) and multiple reaction monitoring (MRM) at m/z [M+H]⁺320.1162.1 for TTX. Result: Calibration curve was linear over the TTX concentration range of 0.0313-2.00 mg·L^-1 (r=0.999 9). The lower limit of quantification of TTX was 0.300 μg·L^-1. The average recovery of TTX was 96.3%-103.0%. Conclusion: The validated method was shown to be simple, sensitive, rapid, reproducible and suitable for the determination of TTX in Fugu liver.