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目的建立免疫亲和柱净化-超高效液相色谱-串联质谱法测定烤鱼片中河豚毒素(TTX)的检测方法。方法样品经酸化甲醇提取,提取液经磷酸盐缓冲液稀释、免疫亲和色谱柱净化后,在ACQUITY UPLC BEH Amide柱(2.1 mm×100 mm,1.7μm)上进行分离,以5 mmol/L乙酸铵溶液(含0.1%甲酸)-乙腈(30∶70,V/V)为流动相,电喷雾正离子多反应模式监测,外标法定量。结果河豚毒素在0.5μg/L~50μg/L时具有良好的线性关系,相关系数>0.995,检出限为0.3μg/kg,回收率为81.7%~92.6%,相对标准偏差<10%。采用该方法对70份烤鱼片样品进行测定,其中9件样品中检出TTX,含量为9.12μg/kg~155μg/kg。结论该方法具有快速、灵敏、简便、准确等优点,适用于烤鱼片中河豚毒素的测定。
OBJECTIVE To establish a method for the determination of tetrodotoxin (TTX) in grilled fish fillets using immunoaffinity column cleanup-ultra performance liquid chromatography-tandem mass spectrometry. Methods The samples were extracted with acidified methanol. The extracts were diluted with phosphate buffer, purified by affinity chromatography, and separated on an ACQUITY UPLC BEH Amide column (2.1 mm × 100 mm, 1.7 μm) with 5 mmol / L acetic acid Ammonium solution (containing 0.1% formic acid) -acetonitrile (30:70, V / V) was used as the mobile phase. Electrospray positive ion was monitored by multiple reaction mode and quantified by external standard method. Results The tetrodotoxin had a good linearity at 0.5μg / L ~ 50μg / L, the correlation coefficient was> 0.995, the detection limit was 0.3μg / kg, the recovery was 81.7% ~ 92.6% and the relative standard deviation was less than 10%. Totally 70 samples of roasted fish fillet were determined by this method. Among them, TTX was detected in 9 samples, the content was 9.12μg / kg ~ 155μg / kg. Conclusion The method is rapid, sensitive, simple, accurate and suitable for the determination of tetrodotoxin in grilled fish fillets.