摘要
基于抗原与抗体间发生特异免疫反应时引起信号变化,结合PAMAM、石墨烯及1-丁基-3-甲基咪唑基四氟硼酸盐3种修饰材料的优势,通过物理方式将黄曲霉毒素B1(Aflatoxin B1,AFB1)抗体固定在修饰电极上建立一种电化学免疫传感器用于食品中AFB1的快速检测。以K_(3)[Fe(CN)_(6)]为探针,采用循环伏安法和差分脉冲伏安法将信号变化可视化,通过研究一定范围内的AFB1浓度与电化学信号间的关系实现对AFB1的定量检测。对试验条件进行了优化,结果为:PAMAM:石墨烯=1:1,(PAMAM:石墨烯):IL=1:1;用量5μL;pH值7.0;孵育温度为37℃;孵育时间30 min。在优化的实验条件下进行研究,发现当AFB1浓度在0.1~8.1 ng/mL之间时,该免疫传感器免疫前后的峰电流变化值与AFB1浓度的对数值呈良好的线性关系,检出限为0.03 ng/mL(S/N=3)。将该方法应用于玉米和玉米油中AFB1的检测,回收率为97.21%~103.02%,检测结果与高效液相色谱法、酶联免疫法均一致,且该法特异性强、稳定性好、重复性高,可以作为食品中AFB1的一种快速检测方法。
An electrochemical immunosensor was constructd for rapid determination of AFB1 in food basing on the signal change caused by the specific immune reaction between antigen and antibody.Immunosensor combined with the advantages of PAMAM,graphene and 1-butyl-3-methylimidazolyl tetrafluoroborate to immobilize aflatoxin B1(AFB1)antibody on the modified electrode physically.K_(3)[Fe(CN)_(6)]was used as a probe,the quantitative detection of AFB1 was realized using cyclic voltammetry and differential pulse voltammetry to visualize the signal changes and study the relationship between AFB1 concentration and electrochemical signal within a certain range.The experimental conditions were optimized as follows:PAMAM:graphene=1:1,(PAMAM:graphene):IL=1:1;Dosage was 5μL;pH=7.0;The incubation temperature was 37℃;The incubation time was 30 min.Under the optimized experimental conditions,the peak current of the immunosensor before and after immunization showed a good linear relationship with the logarithm of AFB1 concentration within 0.1~8.1 ng/mL with limit of detection(LOD)of 0.03 ng/mL(S/N=3).The method was applied to the determination of AFB1 in corn and corn oil,and the recovery rate was 97.21%~103.02%.The detection results were consistent with those of high performance liquid chromatography and enzyme-linked immunosorbent assay.The method can be used as a rapid detection method for AFB1 in food with strong specificity,good stability and high repeatability.
作者
胡俊平
田芳
齐瑞岳
任云
HU Junping;TIAN Fang;QI Ruiyue;REN Yun(Hebei Key Laboratory of Heterocyclic Compounds,College of Chemical Engineering and Materials of Handan University,Handan 056005,China)
出处
《食品科技》
CAS
北大核心
2023年第3期310-318,共9页
Food Science and Technology
基金
邯郸市科学技术研究与发展计划项目(19422072001-15)。
