Aflatoxin B1 Electrochemical Aptasensor Based on Tetrahedral DNA Nanostructures Functionalized Three Dimensionally Ordered Macroporous MoS₂-AuNPs Film

In food safety evaluation, aflatoxin B1 (AFB1) is an important indicator. In this work, we developed an AFB1 electrochemical aptasensor based on a tetrahedral DNA nanostructures (TDNs) immobilized three dimensionally ordered macroporous MoS₂-AuNPs hybrid (3DOM MoS₂-AuNPs) recognition interface and horseradish peroxidase (HRP) functionalized magnetic signal amplifier. To greatly enhance the recognition efficiency, sensitivity, and stability of the aptasensor, the AFB1 aptamer-incorporated TDNs were ingeniously combined with the 3DOM MoS₂-AuNPs film for the construction of the sensing interface. The aptamers would release from the electrode surface after they reacted with AFB1, and then the hybridization-free TDNs formed. Thus, the biocomposite of DNA helper strands (H1)/HRP functionalized AuNPs-SiO₂@Fe₃O₄ nanospheres would combine with the hybridization-free TDNs due to the hybridization of H1 and TDNs. The more AFB1 existed in the solution, the more H1/HRP-AuNPs-SiO₂@Fe₃O₄ could be combined onto the 3DOM MoS₂-AuNPs surface. The current response coming from HRP-catalyzed reduction of H₂O₂ using thionine (Thi) as electrochemical probe was proportional with the AFB1 concentration. Upon optimal conditions, the aptasensor showed specificity for AFB1, achieving a good linear range of 0.1 fg/mL-0.1 μg/mL and the detection limit of 0.01 fg/mL. Furthermore, the developed aptasensor was also applied for detecting AFB1 content in rice and wheat powder samples, obtaining good results in conformity with those achieved from the high-performance liquid chromatography tandem mass spectrometry (HPLC-MS) method.