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目的建立鱼丸中河鲀毒素(TTX)的超高效液相色谱-串联四级杆质谱联用测定方法。方法鱼丸样品中的TTX经酸化甲醇提取,免疫亲和柱净化,氮气吹干后用流动相定容,经Hilic Amide(2.1 mm×100 mm,1.8μm)色谱柱分离,甲醇-0.1%甲酸(含5 mmol/L乙酸铵)溶液作为流动相梯度洗脱,选择电喷雾离子源正离子多反应监测(MRM)模式进行监测,外标法基质标准曲线定量。结果 TTX在HILIC Amide柱上能与杂质良好分离,在2 ng/ml~200 ng/ml时,线性关系良好,相关系数为0.999 1。在10μg/kg、50μg/kg和100μg/kg的加标水平下,回收率为72.9%~95.2%,相对标准偏差为2.32%~5.62%,方法的定量限为10μg/kg。应用该方法对市售的30份鱼丸样品进行检测,均未检出TTX。结论该法操作简单快速,准确度和灵敏度高,适合于鱼丸中TTX的检测,具有较好的应用价值。
Objective To establish a method for determination of tetrodotoxin (TTX) in fish balls by ultra performance liquid chromatography-tandem quadrupole mass spectrometry. Methods TTX in fish sample was extracted by acidified methanol, purified by immunoaffinity column, dried with nitrogen and fixed with mobile phase, separated on a Hilic Amide column (2.1 mm × 100 mm, 1.8 μm), methanol-0.1% formic acid (Containing 5 mmol / L ammonium acetate) as the mobile phase gradient elution. Electrospray ionization (ESI) multi-reaction monitoring (MRM) was used to monitor the concentration. Results TTX was well separated from impurities on the HILIC Amide column with a good linearity of 2 ng / ml ~ 200 ng / ml with a correlation coefficient of 0.999 1. The recoveries ranged from 72.9% to 95.2% at a spiked level of 10μg / kg, 50μg / kg and 100μg / kg with a relative standard deviation of 2.32% -5.62%. The limit of quantification was 10μg / kg. Thirty samples of commercially available fish balls were tested by this method. No TTX was detected. Conclusion The method is simple, rapid, accurate and sensitive, suitable for the detection of TTX in fish balls, and has a good application value.