Due to the diversification and complexity of organophosphorus pesticide residues brings great challenges to the detection work. Therefore, we developed a dual-ratiometric electrochemical aptasensor that could detect malathion (MAL) and profenofos (PRO) simultaneously. In this study, metal ions, hairpin-tetrahedral DNA nanostructures (HP-TDN) and nanocomposites were used as signal tracers, sensing framework and signal amplification strategy respectively to develop the aptasensor. Thionine (Thi) labeled HP-TDN (HP-TDNThi) provided specific binding sites for assembling Pb2+ labeled MAL aptamer (Pb2+-APT1) and Cd2+ labeled PRO aptamer (Cd2+-APT2). When the target pesticides were present, Pb2+-APT1 and Cd2+-APT2 were dissociated from the hairpin complementary strand of HP-TDNThi, resulting in reduced oxidation currents of Pb (I) and Cd (I), respectively, while the oxidation currents of Thi (I) remained unchanged. Thus, I/I and I/I oxidation current ratios were used to quantify MAL and PRO, respectively. In addition, the gold nanoparticles (AuNPs) encapsulated in the zeolitic imidazolate framework (ZIF-8) nanocomposites (Au@ZIF-8) greatly increased the catch of HP-TDN, thereby amplifying the detection signal. The rigid three-dimensional structure of HP-TDN could reduce the steric hindrance effect on the electrode surface, which could greatly improve the recognition efficiency of the aptasensor for the pesticide. Under the optimal conditions, the detection limits of the HP-TDN aptasensor for MAL and PRO were 4.3 pg mL-1 and 13.3 pg mL-1, respectively. Our work proposed a new approach to fabricating a high-performance aptasensor for simultaneous detection of multiple organophosphorus pesticides, opening a new avenue for the development of simultaneous detection sensors in the field of food safety and environmental monitoring.
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