Abstract
将SiO2包覆的Fe3O4磁性纳米材料(SiO2@Fe3O4)表面偶联识别黄曲霉毒素B1(AFB1)的抗体(Ab),用于特异性分离富集谷物中的AFB1,进而与高效液相色谱-串联质谱法(HPLC-MS/MS)结合,用于大米、玉米和小麦中AFB1的高效准确检测。采用微波辅助水热合成法制备得到Fe3O4磁性纳米颗粒,并用100 μL正硅酸乙酯(TEOS)对其进行SiO2的包覆,得到SiO2@Fe3O4磁性纳米材料,随后进行抗体的偶联得到Ab-SiO2@Fe3O4;以pH=7.4的磷酸盐缓冲液(PBS)作为富集缓冲液,加入8 mg Ab-SiO2@Fe3O4,在37 ℃下反应10 min进行AFB1的分离富集,随后采用甘氨酸-盐酸(Gly-HCl)缓冲液对Ab-SiO2@Fe3O4分离富集的AFB1进行洗涤,将洗涤液氮吹后复溶,采用高效液相色谱-串联质谱法检测。在最佳条件下,方法检测AFB1的线性范围为2~50 μg/L,相关系数(R2)>0.99,检出限为0.04 μg/kg,定量限为0.13 μg/kg。在4个不同加标水平下,AFB1在3种谷物基质中的加标回收率为76.21%~92.85%, RSD≤5.29%。大米、玉米和小麦等实际谷物样品中AFB1的测定结果显示,在1个小麦样品和2个玉米样品中检出AFB1,其含量分别为0.38、0.13和0.47 μg/kg,其他样品中并未发现AFB1。方法将磁性纳米材料与HPLC-MS/MS相结合,实现了AFB1的高效分离富集,富集材料成本低廉,储存性能好,在30 min内即可完成前处理过程,可在较短的时间内实现大批量样品的实际分析,在谷物中真菌毒素的检测方面具有良好的应用前景。
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