液质联用快速测定河鲀鱼肝脏中河鲀毒素含量的方法研究

QuEChERS-HPLC-Tandem Mass Spectrometry for Determination of Tetrodotoxin in Fugu Liver

采用高效液相色谱-串联质谱（HPLC-MS/MS）方法进行了河鲀鱼肝脏中河鲀毒素（Tetrodotoxin,TTX）含量的快速检测研究。河鲀鱼肝脏样品用1%乙酸甲醇溶液超声提取2次,正己烷脱脂后用C18和石墨化碳黑（GCB）混合吸附剂净化,冷冻高速离心,上清液经聚四氟乙烯-尼龙复合膜过滤;TSK-gel Amide-80 3μm色谱柱分离,乙腈和0.15%乙酸（含10mmol/L乙酸铵）溶液为流动相梯度洗脱进行HPLC-MS/MS分析。外标法定量测定。方法检出限为40μg/kg,在50～1 000μg/kg范围内线性关系良好。在河鲀鱼肝脏中添加50,75,100μg/kg TTX进行重复性试验,回收率在79.2%～105.3%之间,相对标准偏差（RSD）不大于9.0%（n=6）。本方法准确、灵敏、实用,节约时间和试剂,操作简单,易重现,满足目前对河鲀鱼肝脏中TTX含量快速定量检测的技术要求。

A simple, rapid, sensitive and cheaper high performance liquid chromatography-tandem qua- drupole mass spectrometry（ HPLC-ESI-MS/MS） method was developed for the determination of Tetro- dotoxin（TTX）in Fugu liver. The analytes were extracted from fugu liver samples by acetic acid metha- nol（1 ： 99, V/V）, defatted by hexane then purified by freezed centrifugation and mixture of Cleanert PestiCarb and C18 sobent. The analyte was determined by HPLC-MS/MS using acetonitrile and water （0. 15 % acetic acid and 10 mmol/L ammonium acetate） as the mobile phase by gradient elution. The quantification was performed with the external standard method. The curve of calibration was linear within the range of 50-1000. 0μg/kg and the detection limit was 40μg/kg. Recoveries of spiked tetro- dotoxin at level of 50, 75, 100μ/kg were beteen 79.2 % and 105.3% with relative standard deviation （RSD）was less than 9. 0 %（n=6）. The method has good accuracy, sensitivity, repeatability, simplici- ty, fast and economy to meet practical analysis of TTX in fugu liver samples.