高效液相色谱-串联质谱法检测河豚毒素的 方法研究

Determination of tetrodotoxin by high performance liquid chromatogra-phy-tandem mass spectrometry

中文关键词:  高效液相色谱－串联质谱法  河豚毒素  河豚  亲水性色谱分析柱

英文关键词:high performance liquid chromatography-tandem mass spectrometry  tetrodotoxin  aquatic products  hydrophilic interaction chromatography column

中文摘要:

      目的 建立了一种河豚中河豚毒素(TTX)的高效液相色谱－串联质谱检测方法。方法 取适量样品, 在0.03 mol/L乙酸溶液中均质后超声提取样品中的河豚毒素。粗提液低速离心, 取上清液加入等量甲醇充分混匀后进行高速离心。高速离心后上清液过0.22 μm滤膜后待测。采用Waters ACQUITY UPLC BEH Amide(2.1 mm×100 mm, 1.7 μm)色谱分析柱, 含0.1%甲酸的MilliQ水及乙腈溶液为流动相, 电喷雾正离子模式进行分析。结果 河豚毒素在5~1000 ng/mL的范围内线性关系良好, 相对回收率为83.3%~93.2%, 绝对回收率为47.8%~48.9%, 检出限为5 μg/kg, 定量限为2 0μg/kg。结论 与以往检测TTX的LC-MS/MS方法相比, 新建方法操作简单、快速、准确、灵敏并且实验成本低, 为河豚中TTX的定量检测提供了有效的实用技术手段。

英文摘要:

      Objective To establish a new method for determination of tetrodotoxin (TTX) in puffer fish by high performance liquid chromatography - tandem mass spectrometry. Methods The sample was sonicated after homogenizing with 0.03 mol/L acetic acid solution. After a low-speed centrifugation, the supernatant was mixed with equivalent methanol. With a subsequent high-speed centrifugation, the supernatant was filtered through a 0.22 μm membrane filter before injection. LC-MS/MS analysis was conducted by Waters ACQUITY UPLC BEH Amide (2.1 mm× 100 mm, 1.7 μm) column with the mobile phase of 0.1% formic acid in water and acetonitrile at positive mode. Results There was a good linearity of TTX in the range of 5~1000 ng/mL. The matrix recoveries was 85.8%~93.2% and the absolute recoveries was 47.8%~48.9%. The detection limit is 5 μg/kg and the limit of quantification is 2 0 μg/kg. Conclusion Compared with the previous methods, the new method is simple, rapid, accurate, sensitive and at low cost for the determination of TTX in aquatic products.