超高效液相色谱-串联质谱法测定闽东地区织纹螺中河豚毒素

Determination of tetrodotoxin in Nassarius of eastern Fujian by ultra performance liquid chromatography-tandem mass spectrometry

中文关键词:  河豚毒素  织纹螺  超高效液相色谱-串联质谱法

英文关键词:tetrodotoxin  Nassarius  ultra performance liquid chromatography-tandem mass spectrometry

中文摘要:

      目的 建立一种测定织纹螺中河豚毒素(tetrodotoxin, TTX)含量的超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)。方法 织纹螺样品用1%乙酸水溶液提取, 0.5%乙酸甲醇溶液稀释, -18 ℃冷冻10 min后离心, 经亲水作用色谱柱分离, 乙腈-0.1%甲酸水溶液作为流动相梯度洗脱, UPLC-MS/MS分析, 基质匹配标准曲线外标法定量。结果 该方法的检出限为10 μg/kg, 定量限为25 μg/kg, 满足欧洲食品安全委员会对双壳贝类和腹足类中河豚毒素安全限量值44 μg/kg的检测要求。在织纹螺中添加50、100、250 μg/kg河豚毒素进行加标回收试验, 河豚毒素回收率为79.6%~118.1%, 相对标准偏差为(relative standard deviations, RSDs)为7.7%~10.6% (n=6)。采用该方法测定闽东地区14份织纹螺样品, 最高检出河豚毒素25.7 mg/kg。结论 该研究方法准确性高、成本低、操作性强, 能够满足当前批量定量检测织纹螺中河豚毒素含量的技术要求。

英文摘要:

      Objective To develop a method for the determination of tetrodotoxin (TTX) in Nassarius by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Methods The samples were extracted with 1% acetic acid aqueous solution, diluted with 0.5% acetic acid methanol solution, frozen at -18 ℃ for 10 min, and centrifuged, then separated by hydrophilic column, acetonitrile-0.1% formic acid aqueous solution as mobile phase gradient elution, analyzed by UPLC-MS/MS, and quantified by matrix matching standard curve with external standard method. Results The limit of detection and limit of quantitation were 10 and 25 μg/kg, respectively, which met the current action limit of 44 μg/kg for bivalves and gastropods enforced in the European Food Safety Commission. The method exhibited recoveries ranging from 79.6% to 118.1%, and the relative standard deviations (RSDs) was demonstrated ranging from 7.7% to 10.6% (n=6). This method was used to test 14 batches of Nassarius samples collected from east Fujian, the results of which were found to contain TTX at levels up to 25.7 mg/kg. Conclusion The method has been found to be accurate, low-costed and practical, which can meet the current technical requirements of batch quantitative determination of TTX in Nassarius.