论文部分内容阅读
目的建立高效液相色谱-串联四级杆质谱联用仪多反应监测技术(MRM)测定鱼体中的河豚毒素(TTX)含量的方法。方法海鱼样品用80%酸化甲醇水溶液(含0.1%乙酸)超声提取,二氯甲烷脱脂净化,超滤管离心,取滤液过膜后进行分析。目标物TTX以p H=3.5的乙腈-0.06%甲酸铵溶液(65∶35,V/V)为流动相,经HILIC色谱柱(100 mm×2.1 mm,3μm)等梯度洗脱分离,采用电喷雾离子源,选择质谱多反应监测正离子模式,以保留时间和河豚毒素的二级质谱特征碎片离子予以双定性确证,基质标准曲线校正,外标法定量。结果河豚毒素与杂质能良好分离,在0.10~2.0 mg/kg范围内线性关系良好(r=0.997),方法检出限为0.012 mg/kg,定量限为0.041 mg/kg,加标回收率为75.7%~108.1%,相对标准偏差为1.4%~5.1%。应用该方法对20份实际样品进行检测,均未检出TTX。结论本方法准确、快捷、简便,可应用于食物中毒因子识别与鉴定。
Objective To establish a method for the determination of tetrodotoxin (TTX) in fish by multi-reaction monitoring (MRM) with high performance liquid chromatography-tandem quadrupole mass spectrometry. Methods The marine fish samples were extracted with 80% acidified aqueous solution of methanol (containing 0.1% acetic acid). The samples were degreased with methylene chloride and centrifuged in ultrafiltration tubes. The filtrate was passed through the membrane and analyzed. The target TTX was separated on a HILIC column (100 mm × 2.1 mm, 3 μm) using acetonitrile-0.06% ammonium formate solution (65:35, V / V) with p H = 3.5 as mobile phase. Spray ion source, select the mass spectrometry multiple reaction monitoring positive ion mode, retention time and tetrodotoxin secondary ion mass spectrometry fragment ions to be double qualitative confirmation, matrix calibration curve calibration, external standard method. The results showed that tetrodotoxin could be separated from impurities with good linearity in the range of 0.10-2.0 mg / kg (r = 0.997). The detection limit was 0.012 mg / kg and the limit of quantification was 0.041 mg / kg. The recoveries were 75.7% ~ 108.1%, the relative standard deviation was 1.4% ~ 5.1%. Twenty actual samples were tested by this method and no TTX was detected. Conclusion The method is accurate, rapid and simple and can be applied to the identification and identification of food poisoning agents.