基于丝网印刷电极和重氮官能化的赭曲霉素A电化学免疫传感检测方法的建立与应用  

Establishment and Application of Ochratoxin A Detection Immunosensor Method Based on Screen Printing Electrodes and Diazo Functionalization

作　　者：宋心怡 王鑫[1] 赵晓磊[1] 何金兴[1] SONG Xinyi;WANG Xin;ZHAO Xiaolei;HE Jinxing(Qilu University of Technology(Shandong Academy of Sciences),Jinan 250000,Shandong,China)

机构地区：[1]齐鲁工业大学(山东省科学院),山东济南250000

出　　处：《食品研究与开发》2024年第17期149-158,共10页Food Research and Development

基　　金：国家自然科学基金项目(32102061);山东省自然科学基金面上项目(ZR2021MC187)。

摘　　要：该文将赭曲霉素A(ochratoxin A,OTA)抗体和重氮盐接枝在电极表面,构建一种用于OTA快速检测的电化学免疫传感器。通过电沉积的方法将重氮盐修饰于丝网印刷碳电极(screen-printed carbon electrodes,SPCE),接枝的重氮膜作为底层,使羧基官能团暴露在电极表面。然后使用碳二亚胺对羧基进行活化,用交联的方式使抗体和羧基进行酰胺键合从而实现抗体的固定化。利用循环伏安(cyclic voltammetry,CV)法和电化学阻抗谱(electrochemical impedance spectroscopy,EIS)表征电化学免疫传感器的特性。结果表明,在最佳工作条件下,所开发的电化学免疫传感器的线性检测范围为20~200 ng/mL(R2=0.9970),检测限为0.5 ng/mL,在检测加标样品中OTA方面表现出优异的电化学性能,适用于食品中OTA的检测。此外,在干扰物存在的情况下,该传感器对OTA具有高度选择性,且贮存14 d稳定性仍较好。该电化学免疫传感器易于构建、灵敏、快速且稳定,对实际样品中OTA的检测限低,分析时间短和成本低。In this paper,an electrochemical immunosensor for the rapid detection of ochratoxin A(OTA)was constructed by grafting OTA antibody and diazonium salt on the electrode surface.In this work,the screenprinted carbon electrodes(SPCE)were modified with diazo salt by electro-deposition.The grafted diazo film served as the bottom layer to expose the carboxyl functional groups to the electrode surface.Then carbodiimide was used to activate the carboxyl groups.The antibody and carboxyl groups were crosslinked to conduct amide bonding to realize the immobilization of the antibody.The electrochemical immunosensors were characterized by cyclic voltammetry(CV)and electrochemical impedance spectroscopy(EIS).It was found that under the best working conditions,the linear detection range of the developed electrochemical immunosensor was 20-200 ng/mL(R2=0.9970),and the detection limit was 0.5 ng/mL.Showing excellent electrochemical performance in the detection of OTA in labeled samples,the immunosnesor was suitable for the detection of OTA in food.In addition,in the presence of interferent,the sensor was highly selective to OTA and stayed stable for 14 days.The electrochemical immunosensor is easy to construct,sensitive,fast,and stable,with low detection limits,analysis time,and cost for OTA in real samples.

关 键 词：赭曲霉素A 电化学免疫传感器 重氮化 丝网印刷碳电极 快速检测 

分 类 号：O657.1[理学—分析化学] TS207.5[理学—化学]