基质分散固相萃取净化-亲水液相色谱-串联质谱法检测织纹螺与贝类中河鲀毒素

Determination of tetrodotoxin in nassarius and shellfish by hydrophilic liquid chromatography-tandem mass spectrometer coupled with dispersive solid phase extraction

目的针对海产品常见中毒原因分析需求,建立基质分散固相萃取净化-亲水液相色谱-串联三重四极杆质谱(HILIC-MS/MS)快速定性定量检测织纹螺和贝类中河鲀毒素的新方法。方法 1.0 g样品经0.1%乙酸溶液沸水浴提取后,用50 mg亲水亲油平衡填料(HLB)和5 mg石墨化碳黑(GCB)吸附剂吸附净化,最后经乙腈蛋白沉淀后过0.22μm聚四氟乙烯(PTFE)滤膜,亲水液相色谱柱(150 mm×2.0 mm,3μm)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2.0~40.0 ng/ml浓度范围内,河鲀毒素呈现良好的线性关系,相关系数r^2≥0.999;以3倍基线噪声所对应的浓度为检出限时,河鲀毒素的方法检出限可达10.0μg/kg;在25、100和200μg/kg的加标水平时,方法回收率为74.2%~87.9%,相对标准偏差为2.3%~9.1%。应用本方法对浙江沿海地区市售织纹螺和贝类样品进行检测,15份织纹螺中有14份检出河鲀毒素,检出率为93.3%,含量范围为0.04~15.75 mg/kg,60份贝类样品均未检出河鲀毒素。结论该检测方法准确、快速、易操作,能满足典型海产品中河鲀毒素的公共卫生应急检测或日常监测要求。

Objective In order to analyze of poisoning causes,a new method was established utilizing hydrophilic liquid chromatography-tandem triple quadrupole mass spectrometer（ HILIC-MS/MS） coupled with dispersive solid phase extraction for rapid qualitative and quantitative analysis of tetrodotoxin in nassarius and shellfish. Methods Sample（ 1. 0 g） was extracted with 0. 1% acetic acid in boiling water bath,purified by dispersive solid phase extraction with50 mg hydrophilic-lipophilic balance（ HLB）,5 mg graphitized carbon black（ GCB） and protein precipitation with acetonitrile,and then filtered through a polytetrafluoroethylene（ PTFE） membrane. The analytes were 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 quantification. Results Tetrodotoxin showed good linearity in the concentration range between 2. 0 and 40. 0 ng/ml,the correlation coefficient was higher than 0. 999. The detection limit of tetrodotoxin in seafood was 10. 0 μg/kg. The rates of recovery varied between 74. 2% and 87. 9% with relative standard deviations from 2. 3% to 9. 1% at spiked concentrations of 25,100 and 200 μg/kg. The proposed method was applied in the detection of tetrodotoxin in shellfish and nassarius from coastal cities of Zhejiang Province. Conclusion The method was accurate,fast,easy to operate,which could meet the requirements of public health emergency testing or routine testing.