QuEChERS/液相色谱-串联质谱法测定红鳍东方鲀肉中河鲀毒素

Determination of Tetrodotoxin in Flesh of Fugu Rubripes by QuEChERS Combined with Liquid Chromatography-Tandem Mass Spectrometry

建立了快速检测红鳍东方鲀肉中剧毒生物毒素河鲀毒素（Tetrodotoxin,TTX）含量的高效液相色谱-串联质谱（HPLC-MS/MS）确证方法.分析物用1％乙酸-甲醇溶液于50℃超声提取,C18和石墨化碳黑（GCB）混合粉末作为吸附剂QuEChERS净化,高速冷冻离心,上清液经尼龙-聚四氟乙烯复合膜过滤;TSK-gel Amide-80 3 μm色谱柱分离,乙腈-0.15％乙酸（含10 mmol/L乙酸铵）溶液作为流动相梯度洗脱,对滤液进行HPLC-MS/MS分析.外标法质标准曲线定量.

A simple, rapid, sensitive and cheap high performance liquid chromatography -tandemquadrupole mass spectrometric （HPLC - MS/MS） method was developed for the determination of tetro-dotoxin（Trx） that is one of the most toxic non protein small biotoxin in Fugu rubripes muscle. Theanalytes were ultrasonically extracted from homogeneous fugu muscle samples with 1% acetic acidmethanol solution at 50 ℃, purified by freezing high speed centrifugation, and succeeded cleaned byQuEChERS （ Quick, Easy, Cheap, Effective, Rugged and Safe） method using a mixture of CleanertPestiCarb and Cls as sorbent. The analyte was determined by HPLC - MS/MS using acetonitrile and0. 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 stand-ard method. Recoveries of TYX at spiked levels of 30, 50, 100 μg/kg were between 74. 0% and105.0% with relative standard deviations（ RSD, n=6） not more than 11.0%. The limit of detec-tion and limit of quantitation were 10 /xg/kg and 30 μg/kg, respectively. With the advantages ofgood accuracy, repeatability, sensitivity, simplicity, economy and environment friendiness, thismethod could meet the requirements of practical analysis of Trx in fugu flesh samples.