亲和免疫柱净化-高效液相色谱-三重四级杆串联质谱法测定土鳖虫中的黄曲霉毒素

Detection of aflatoxins in Eupolyphaga by immunoaffinity column clean-up and high performance liquid chromatography with triple quadrupole mass spectrometry

目的建立土鳖虫中的黄曲霉毒素(AF)的亲和免疫柱净化-高效液相色谱-三重四级杆串联质谱检测方法。方法以X-Brigde-C18(3.5μm,3.0 mm×50 mm)色谱柱分离,串联四级杆质谱仪检测,MRM模式进行定性定量分析,样品以60%甲醇为溶剂提取,检测土鳖虫中的AF B1、AF B2、AF G1、AF G2。结果 4种AF质谱检测的线性范围宽,相关性好,r≥0.997 9;方法精密度以6次测定值的RSD表示,为1.5%~3.4%;方法回收率用3个浓度进行添加实验,回收率范围为85.6%~98.9%;定量限范围为0.2μg/L^0.7μg/L;日内精密度的RSD(n=5)范围为1.1%~2.9%,日间精密度的RSD(n=9)范围为1.5%~2.8%。结论本方法专属性强,操作简单,快捷,可作为土鳖虫中AF定量定性的有效检测方法。

Objective To develop an immunoaffinity column clean-up and high performance liquid chromatography with triple quadrupole mass spectrometry（ HPLC-MS/MS） method for the determination of aflatoxins（ AF） in Eupolyhpaga.Methods The analysis was performed by a HPLC-MS/MS system with X-Brigde-C18（ 3. 5 μm,3. 0 mm × 50 mm） column.Multiple-reaction monitoring（ MRM） was performed to identify and quantify AF B1,AF B2,AF G1 and AF G2,which were extracted with 60% methanol-water. Results Four linear calibration curves were obtained with good correlation,with r≥0. 997 9.The precision of the method was showed by RSD（ n = 6） ranged from 1. 5% to 3. 4%. The recoveries were determined at three concentrations and ranged from 85. 6% to 98. 9%. The ranges of LOQ were from 0. 2 μg/L to 0. 7 μg/L and the RSDs of intra-day precision（ n = 5） and inter-day precision（ n = 9） were within 1. 1%-2. 9% and 1. 5%-2. 8%,respectively. Conclusion The method is specific,simple and fast and can be used to detect aflatoxins in Eupolyhpaga.