超声-固相萃取高效液相色谱-串联质谱法测定红鳍东方豚中的河豚毒素

Determination of tetrodotoxin in Fugu rubripes by high performance liquid chromatography-tandem mass spectrometry coupled with ultrasonic extraction and solid phase extraction

中文关键词:  高效液相色谱-串联质谱法  红鳍东方豚  河豚毒素

英文关键词:high performance liquid chromatography-tandem mass spectrometry  Fugu rubripes  tetrodotoxin

中文摘要:

      目的 建立快速、准确检测红鳍东方豚中河豚毒素(tetrodotoxin, TTX)的高效液相色谱-串联质谱(HPLC-MS/MS)的分析方法。方法 样品用0.1%乙酸水溶液超声提取, C18固相萃取柱净化, 用亲水性色谱柱ZIC?-cHILIC(150 mm×2.1 mm, 3 μm)洗脱, 乙腈-0.1%甲酸水为流动相。选择正离子多反应监测(MRM)模式, [M＋H]+, m/z 320.3/302.2为定量离子, 进行HPLC-MS/MS 测定, 外标法定量。结果 河豚毒素在5~100 ng/mL的范围内呈良好的线性关系, 相关系数为0.9997, 回收率为82%~91%, 相对标准偏差RSD为 5.3%~15.7%, 方法的定量限为5 ng/mL。结论 该方法快速准确, 灵敏度高, 适用于红鳍东方豚中河豚毒素的测定。

英文摘要:

      Objective To establish an accurate, rapid analytical high performance liquid chromatography- tandem mass spectrometry (HPLC-MS/MS) method for the determination of tetrodotoxin (TTX) in the Fugu rubripes. Methods TTX was extracted from sample by 0.1% acetic acid-water, and then purified by solid-phase extraction with C18 sorbent. The separation was performed on the ZIC?-cHILIC (150 mm×2.1 mm, 3 μm) at 35 ℃ using acetonitrile-0.1% acetic acid-water(90:10, v:v) as mobile phase. Samples were analyzed by HPLC-MS/MS and quantified with the external standard method. Quantification was carried out through tandem mass spectrometry with positive electro-spray ionization (ESI) and multiple reaction monitoring (MRM) at m/z [M+H] 320.3 302.2 for TTX. Results Calibration curve was linear over the TTX concentration range of 5~100 ng/mL (r=0.9997), with the limit of quantification of 5 ng/mL. The recoveries were between 82% and 91% with the relative standard deviations (RSD) ranging from 5.3% to 15.7%. Conclusion The method was shown to be sensitive, rapid and suitable for the determination of TTX in Fugu rubripes.