†Key Lab of Sustainable Utilization of Technology Research for Fishery Resource of Zhejiang Province, Marine Fisheries Research Institute of Zhejiang, 28 Tiyu Street, Zhoushan 316021, P.R. China.
‡Jiangsu Meizheng Biotechnology Company Limited, Wuxi 214135, P.R. China.
J Agric Food Chem. 2015 Apr 1;63(12):3129-34. doi: 10.1021/acs.jafc.5b00045. Epub 2015 Mar 23.
A highly selective and sensitive method was developed for the determination of tetrodotoxin (TTX) in marine organisms by immunoaffinity chromatography (IAC) purification coupled with ultrahigh performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). An IAC column was prepared and used to cleanup the extracted samples. The operating conditions of the IAC column were optimized, and the capacity of new IAC column was found to be 1106 ng mL(-1), which was sufficient for TTX determination. The MS/MS conditions and UPLC mobile phase were also studied to optimize the operation conditions. Fortified marine organism samples at levels of 0.3-5.0 ng g(-1) were utilized, and the average recoveries were 86.5-103.6% with intra- and inter-day relative standard deviations less than 7.22 and 9.88%, respectively. The limits of detection and quantification were 0.1 and 0.3 ng g(-1), respectively. The method was later successfully applied for the determination of TTX in 100 marine organism samples collected from local markets.
建立了一种用于通过免疫亲和色谱（IAC）纯化结合超高效液相色谱串联质谱（UPLC-MS/MS）测定海洋生物中河豚毒素（TTX）的高度选择性和灵敏的方法。制备了IAC 柱并用于净化提取的样品。优化了 IAC 柱的操作条件，发现新的 IAC 柱的容量为 1106ngmL(-1)，足以进行 TTX 测定。还研究了 MS/MS 条件和 UPLC 流动相，以优化操作条件。利用水平为 0.3-5.0ngg(-1)的加标海洋生物样品，平均回收率为 86.5-103.6%，日内和日间相对标准偏差分别小于 7.22%和 9.88%。检测限和定量限分别为 0.1 和 0.3ngg(-1)。该方法后来成功应用于从当地市场采集的 100 个海洋生物样品中测定 TTX。
