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目的采用免疫亲和柱净化鱼肉和肝脏中的河鲀毒素,建立高效液相色谱-三重四级杆质谱串联(LC-MS/MS)方法检测鱼肉和肝脏中的河鲀毒素,为水产品中的河鲀毒素检测提供方法依据。方法选用Zic-Hilic色谱柱(150 mm×2.1 mm,5μm),以10 mmol/L甲酸铵-0.1%甲酸-乙腈为流动相,采用梯度洗脱进行分离。样品用1%乙酸-甲醇沉淀蛋白提取,上清液加入PBS缓冲液后经免疫亲和柱净化,将洗脱液氮吹至干定容后上机测定。多重反应监测(MRM)方式检测。结果河鲀毒素的线性范围为1.0~1 000.0 ng/ml,鱼肉和肝脏中河鲀毒素的检出限分别为0.3和0.2μg/kg,回收率在52.4%~72.6%之间。结论本方法特异性强、提取效果好、无基质抑制效应,适用于鱼肉和肝脏中河鲀毒素的痕量检测。
Objective To purify tetrodotoxin in fish and liver by immunoaffinity column and establish the method of high performance liquid chromatography-triple quadrupole mass spectrometry (LC-MS / MS) to detect tetrodotoxin in fish and liver, The method of detection of tetrodotoxin is provided. Methods Zic-Hilic column (150 mm × 2.1 mm, 5 μm) was used as the mobile phase. The mobile phase consisted of 10 mmol / L ammonium formate-0.1% formic acid-acetonitrile and separated by gradient elution. Samples were extracted with 1% acetic acid-methanol precipitated protein. The supernatant was added to PBS buffer and purified by immunoaffinity column. The eluate was blown to dryness and then measured on the machine. Multiple Reaction Monitoring (MRM) mode detection. Results The linear range of tetrodotoxin was 1.0-1 000.0 ng / ml. The detection limits of tetrodotoxin in fish meat and liver were 0.3 and 0.2 μg / kg, respectively. The recoveries ranged from 52.4% to 72.6%. Conclusion The method has strong specificity, good extraction effect and no matrix inhibition effect, and is suitable for the trace detection of tetrodotoxin in fish and liver.