A simple,rapid,sensitive and cheap high performance liquid chromatography-tandem quadrupole mass spectrometric（HPLC-MS/MS） method was developed for the determination of tetrodotoxin（TTX）that is one of the most toxic non protein small biotoxin in Fugu rubripes muscle.The analytes were ultrasonically extracted from homogeneous fugu muscle samples with 1% acetic acid methanol solution at 50 ℃,purified by freezing high speed centrifugation,and succeeded cleaned by QuEChERS（Quick，Easy，Cheap，Effective，Rugged and Safe） method using a mixture of Cleanert PestiCarb and C18 as sorbent.The analyte was determined by HPLC-MS/MS using acetonitrile and 0.15% acetic acid（containing 10 mmol/L ammonium acetate）as mobile phase by gradient elution．The quantitation of TTX was performed using matrix-matched calibration curve by the external standard method.Recoveries of TTX at spiked levels of 30,50,100 μg/kg were between 74.0% and 105.0% with relative standard deviations（RSD,n=6） not more than 11.0%.The limit of detection and limit of quantitation were 10 μg/kg and 30 μg/kg,respectively.With the advantages of good accuracy,repeatability,sensitivity,simplicity,economy and environment friendiness,this method could meet the requirements of practical analysis of TTX in fugu flesh samples．