液相色谱串联质谱法检测腐乳中的黄曲霉毒素

Determination of aflatoxins in Sufu by ultra-high performance liquid chromatography tandem mass spectrometry

目的建立UPLC-MS/MS检测腐乳中4种黄曲霉毒素的方法。方法样品用乙腈/水(86/14,V/V)混合匀浆超声提取10 min,高速离心,上清液过多功能净化柱MycosepTM226,氮气吹至近干,初始流动相定容,0.22μm微孔滤膜过滤,经过Agilent ZORBAX XDB C18色谱柱分离,以乙腈和0.1%甲酸溶液为流动相,梯度洗脱,进行UPLC-MS/MS分析。质谱采用正离子扫描模式,离子源为ESI(+),定量检测方式为多反应监测(MRM)方式,利用保留时间和碎片信号比值判定定性结果。结果方法的线性范围为0.001μg/kg^0.16μg/kg,线性相关系数均>0.999 0,检出限为0.001μg/kg^0.009μg/kg;低、中、高3个水平的平均加标回收率为96.4%~99.7%,相对标准偏差(RSD)均<2.0%。结论该方法用于测定腐乳中4种黄曲霉毒素具有操作简单、干扰少、快速、准确可靠等特点,可作为腐乳中黄曲霉毒素检测的确证方法。为市场监督提供良好的基础数据。

Objective To establish a new method for quantitative analysis of aflatoxins in sufu using ultra-high performance liquid chromatography tandem mass spectrometry（ UPLC-MS/MS）. Methods The homogenized samples were extracted by acetonitrile aqueous solution（ 84/16,V/V） for 10 min in super-sonic instrument. After high speed centrifugation,it was purified by MycosepTM226 multifuntional column. Then the purified solution was concentrated by nitrogen,dissolved with the initial mobile phase and cleaned through 0. 22 μm millipore filter membrane. It was separated on a Agilent ZORBAX XDB C18 column with acetontril and water（ containing 0. 1% formic acid） as mobile phase for gradient elution,finally analyzed with UHPLC-MS/MS. Analyte identification and quantification were performed using multiple raction monitoring（ MRM） and electrospray ionization in positive. The ratio of retention time and fragment signal was used for qualitative analysis. Results The linearity range of this method was within 0. 001 μg/kg-0. 16 μg/kg,with correlation coefficients greater than 0. 999 0. The detection limit of this method was within 0. 001 μg/kg-0. 009 μg/kg; the average rcoveries of low,mid,high three spiking levels were within 96. 4%-99. 7% and the RSDs was less than 2. 0%. Conclusion The method of determination of 4 aflatoxins in sufu by UPLC-MS/MS had the advantages of easy operation,less interference from impurities and good accuracy. It could be used for determining 4 aflatoxins in sufu products,and provide basic data for market supervision.