分散固相萃取净化-液质联用法快速检测尿液中河豚毒素  

Rapid determination of tetrodotoxin in human urine by liquid chromatography-tandem mass spectrometer coupled with dispersive solid phase extraction

作　　者：方力[1] 邱凤梅 陈传 FANG Li;QIU Feng-mei;CHEN Zhuan(Key Laboratory of Health Risk Factors for Seafood of Zhejiang Prouince Zhoushan Municipal District Cenler for Disease Control and Preuention Zhoushan,Zhejiang 316021,China;不详)

机构地区：[1]浙江省海产品健康危害因素关键技术研究重点实验室,舟山市疾病预防控制中心,浙江舟山316021 [2]浙江省舟山市普陀区疾病预防控制中心,浙江舟山316100 [3]杭州医学院检验医学院,浙江杭州311399

出　　处：《中国卫生检验杂志》2022年第12期1434-1438,共5页Chinese Journal of Health Laboratory Technology

基　　金：舟山市科技计划项目(2021C31071);舟山市医药卫生科技计划项目(2020YA01)。

摘　　要：目的建立分散固相萃取净化-液相色谱-串联质谱快速定性定量检测尿液基质中河豚毒素的方法。方法准确吸取1.0ml尿液,用50mg亲水亲脂平衡(HLB)填料和2.5mg石墨化碳(GCB)吸附剂净化后过0.22μm PTFE滤膜,Acquity UPLC BEH Amide柱(150mm×3.0mm,1.7μm)分离,电喷雾离子化,选择反应监测模式检测,基质匹配外标法定量。结果在1.0μg/L~100μg/L浓度范围内,河豚毒素呈良好的线性关系,相关系数≥0.999;以10倍基线噪声所对应的浓度为定量限时,方法定量限可达1.0μg/L;在2.0μg/L,5.0μg/L、20.0μg/L和50.0μg/L的加标水平时,方法的日内和日间回收率分别为89.3%~94.4%和93.5%~99.0%,日内与日间相对标准偏差均<7.1%。结论该检测方法准确、快速成本低、易操作,适用于中毒患者尿样中河豚毒素的公共卫生应急检测和临床中毒病因识别检测。Objective To establish a method for quantitative analysis of tetrodotoxin in human urine using liquid chromatography-tandem mass spectrometer(LC-MS/MS) coupled with dispersive solid phase extraction.Methods A total of 1. 0 ml urine sample was cleaned up with dispersive solid phase extraction with 50 mg hydrophile-lipophile balance(HLB) material and 2. 5 mg graphitizable carbon(GCB), and then filtered through a PTFE membrane. The analyte was separated on an Acquity UPLC BEH Amide column(150 mm × 3. 0 mm, 1. 7 μm), 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 in the concentration range between 1. 0 μg/L-50. 0 μg/L, with correlation coefficient higher than0. 999. The quantitation limit of tetrodotoxin in human urine was 5 μg/L. The inter-and intra-day recoveries were 89. 3%-94. 4% and 93. 5%-99. 0%, respectively. The inter-and intra-day relative standard deviations(RSDs) were lower than7. 1% at spiked concentrations of 2. 0 μg/L, 5. 0 μg/L, 20. 0 μg/L, and 50. 0 μg/L.Conclusion The method is accurate,fast, easy to operate, which can satisfy the requirements of public health emergency testing or clinical testing.

关 键 词：分散固相萃取 尿液 河豚毒素 液相色谱-串联质谱法 中毒 

分 类 号：O657[理学—分析化学]