亲水液相色谱-串联质谱法检测即食烤鱼片中的河豚毒素

Determination of tetrodotoxin in instant roast fish fillet by hydrophilic interaction chromatography-tandem mass spectrometer

目的针对即食烤鱼片中毒原因分析这一迫切需求,利用亲水液相色谱-串联三重四极杆质谱仪(HILIC-MS/MS)建立了快速定性定量检测即食烤鱼片中河豚毒素的方法。方法取1.0 g即食烤鱼片样品经0.1%甲酸溶液提取,HLB固相萃取柱净化后过膜,亲水液相色谱柱分离,电喷雾离子化,选择反应监测(SRM)模式检测,基质匹配外标法定量。结果在2.5 ng/ml^100 ng/ml时河豚毒素呈现良好的线性关系,相关系数为0.999 5;以3倍基线噪声所对应的浓度为检出限时,即食烤鱼片基质中河豚毒素的方法检出限可达10.0μg/kg;在25μg/kg、100μg/kg和200μg/kg的加标水平时,方法的回收率为73.4%~89.5%,相对标准偏差为2.9%~10.4%。结论该检测方法准确、快速、易操作,能满足即食烤鱼片中河豚毒素的公共卫生应急检测或日常监测要求。

Objective In order to satisfy the causes analysis of instant toast fish fillet poisoning,a new method utilizing hydrophilic interaction chromatography- tandem triple quadrupole mass spectrometer（ HILIC- MS / MS） for rapid qualitative and quantitative analysis of tetrodotoxin in instant roast fish fillet was established. Methods 1. 0 g instant toast fish fillet samples was extracted with 0. 1% formic acid,HLB solid phase extraction column was used to purify and then filtered by membrane.The analyte was separated on a HILIC column,and detected in selected reaction monitoring（ SRM） mode via positive electrospray ionization. The matrix matching and external standard method was used for quantitation. Results Tetrodotoxin showed good linearity with correlation coefficient of 0. 999 5 in the concentration range between 2. 5 ng / ml and 100 ng / ml. The detection limit of tetrodotoxin in instant toast fish fillet can reach to 10. 0 μg / kg based on the 3- time baseline noise. The rates of recovery varied between 73. 4% and 89. 5% with relative standard deviations from 2. 9% to 10. 4% at spiked concentrations of25 μg / kg,100 μg / kg and 200 μg / kg. Conclusion The method is accurate,fast,easy to operate,which can satisfy the requirements of public health emergency testing or routine testing.