亲水液相色谱-三重四极杆质谱检测河豚鱼肝中的河豚毒素

Determination of Tetrodotoxin in Fugu Liver Using Hydrophilic Interaction Liquid Chromatography Coupled with Triple Quadrupole Mass Spectrometry

目的:建立河豚鱼肝中河豚毒素(TTX)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。极性聚酰胺键合硅胶柱InnovationTMHP Amide(100 mm×3.0mm,5μm),流动相乙腈-0.3%甲酸(体积比为70∶30),流速为0.5 mL·min-1;柱温30℃,进样量5μL。选择正离子多反应监测(MRM)模式,[M+H]+,m/z 320.1162.1进行定量分析。结果:TTX的线性范围为0.031 3～2.00 mg·L-1(r=0.999 9);检测限(S/N=3)为3.0 pg,加样回收率在96.3%～103.0%。结论:该法灵敏度高,重复性好,简便快速,适用于河豚毒素的测定。

Objective：To develop a LC-MS-MS method for determination of tetrodotoxin（TTX） in Fugu liver.Method：The analyte was vortex-mixed,extracted with acetonitrile by ultrasound,and followed by centrifugation.The supernatant（5 μL） 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 mobile phase at a flow rate of 0.5 mL·min-1.Quantification was through tandem-mass spectrometry with positive electro-spray ionization（ESI） and multiple reaction monitoring（MRM） at m/z ＋320.1162.1 for TTX.Result：Calibration curve was linear over the TTX concentration range of 0.0313-2.00 mg·L-1（r=0.999 9）.The lower limit of quantification of TTX was 0.300 μg·L-1.The average recovery of TTX was 96.3%-103.0%.Conclusion：The validated method was shown to be simple,sensitive,rapid,reproducible and suitable for the determination of TTX in Fugu liver.