高效液相色谱-串联质谱法检测腰果中的黄曲霉毒素

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

建立了腰果中4种黄曲霉毒素的高效液相色谱-串联质谱检测方法(HPLC-MS/MS)。样品用甲醇-水(8∶2,v/v)溶液提取后用弗罗里硅土柱净化,5 mL丙酮-水-甲酸溶液(96∶3.5∶0.5,v/v/v)洗脱,氮吹至干,1 mL甲醇定容;在资生堂MG C18色谱柱(100 mm×3.0 mm,3μm)上梯度洗脱分离,然后采用电喷雾离子化三重四极杆串联质谱测定。实验结果表明,4种黄曲霉毒素在各自的线性范围内峰面积与其质量浓度线性关系良好,相关系数(r2)大于0.997;检出限(信噪比为3)为0.009～0.04μg/kg,定量限(信噪比为10)为0.03～0.12μg/kg;平均回收率为63.0%～78.5%,相对标准偏差为2.8%～9.1%,均符合痕量分析的要求。评价了基质效应,信号抑制/增强值为88.8%～99.4%,说明净化后的基质效应较小。该方法简单快速、准确可靠,可用于腰果中黄曲霉毒素的检测。

A method for the determination of four aflatoxins in cashew using high performance liquid chromatography-tandem mass spectrometry（HPLC-MS/MS） was developed.The sample was extracted with methanol-water（8∶2,v/v） solution,followed by a cleanup procedure with Florisil column.The target compounds were eluted using 5 mL acetone-water-formic acid（96∶3.5∶0.5,v/v/v） solution.The eluate was dried under N2,then dissolved in 1 mL methanol.Four aflatoxins were separated in MG C18 column（100 mm×3.0 mm,3 μm） adopting a gradient program within 15 min.A triple quadrupole mass spectrometry equipped with an electrospray ionization source operated in the positive ion mode was used to detect the aflatoxins.The good correlation coefficients（r20.997） of the four aflatoxins were obtained within their respective linear ranges.The limits of detection（S/N=3） were between 0.009 μg/kg and 0.04 μg/kg,and the limits of quantification（S/N=10） were between 0.03 μg/kg and 0.12 μg/kg.The recoveries were in a range of 63.0%-78.5% with the relative standard deviations（RSDs） varied from 2.8% to 9.1%.The validation results meet the requirements of trace assay.Matrix effects were estimated and the signal suppression/enhancement ranged from 88.8% to 99.4%.The results indicate that the developed method is simple,fast,accurate,and can be applied for the determination of fours aflatoxins in cashew.