亲水液相色谱-质谱联用法测定河豚子中的河豚毒素

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,5 μm);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5 μL。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0.0313~2.00 μg·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,5 μm)at 30 ℃ using acetonitrile-0.3% formic acid (70∶30)as the mobile phase at a flow rate of 0.5 mL·min^-1.Quantification was performed by tandem-mass spectrometry with positive 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 recovery of tetrodotoxin was 95.9%-102.7%. Conclusion: The validated method is shown to be simple,sensitive,rapid,reproducible and suitable for the determination of tetrodotoxin in Fugu roe.