亲水液相色谱-质谱联用法测定河豚子中的河豚毒素

Determination of tetrodotoxin in Fugu roe by hydrophilic interaction liquid chromatography coupled with triple quadrupole mass spectrometry

目的:建立河豚子中河豚毒素的亲水液相色谱三重四极杆质谱联用(LC-MS/MS)定量检测方法。方法:样品经匀浆,超声,乙腈沉淀离心后取上清液进样测定。色谱柱为Innovation HP Amide柱(100 mm×3.0 mm,5μm);流动相为乙腈-0.3%甲酸(体积比为70∶30);流速为0.5 mL.min-1;柱温为30℃;进样量5μL。以正离子多反应监测(MRM)方式进行定量分析,用于监测的离子对为[M+H]+320.1→162.1。结果:河豚毒素的线性范围为0.0313～2.00μg.mL-1,r=0.9999;检测限(S/N=3)为0.30 ng.mL-1,即3.0 pg;加样回收率在95.9%～102.7%之间。结论:本法简单、快速,灵敏度高,重复性好,适用于河豚子中河豚毒素的定量测定。

Objective： To establish an LC - MS/MS method for the determination of tetrodotoxin in Fugu roe. Methods： The analyte was vortex - mixed, extracted with acetonitrile by ultrasound followed by centrifugation. The supernatant was injected into the LC - MS/MS system for analysis. The separation was performed on an Innovation HP Amide column （100 mm×3.0 mm,5μm）at 30 ℃ using acetonitrile -0. 3 % formic acid （70： 30）as the mo-bile phase at a flow rate of 0. 5 mL · min^ - 1. Quantification was performed by tandem - mass spectrometry with posi- tive electro - spray ionization （ESI） and multiple reaction monitoring （MRM） at [ M ＋ H] ^＋ m/z 320. 1→162. 1 for tetrodotoxin. Results：The calibration curve was linear over the tetrodotoxin concentration range of 0. 0313 -2.00 μg · mL^ - 1 （ r = 0. 9999）. The lower limit of quantification of tetrodotoxin was 0. 30 ng · mL^ - 1. The average recov-ery of tetrodotoxin was 95.9% - 102. 7%. Conclusion：The validated method is shown to be simple, sensitive, rap-id, reproducible and suitable for the determination of tetrodotoxin in Fugu roe.