Visual electrochemiluminescence biosensing of aflatoxin M1 based on luminol-functionalized, silver...

A sensitive electrochemiluminescence (ECL) aptasensor for aflatoxin M1 (AFM1) detection by a closed bipolar electrode (BPE) array has been introduced. The thiolated AFM1 aptamer was immobilized on gold nanoparticle-coated magnetic Fe₃O₄ nanoparticles (Apt-GMNPs). Luminol-functionalized silver nanoparticle-decorated graphene oxide (GO-L-AgNPs) participates in π-π interactions with the unpaired bases of the immobilized aptamer (Apt-GMNPs-GO-L-AgNPs). After the Apt-GMNPs-GO-L-AgNPs were introduced to a gold anodic BPE array, the individual electrodes were subjected to different concentrations of AFM1. Upon the interaction of AFM1 with the aptamers, the GO-L-AgNPs detach from the aptamer; the resulting ECL of luminol and H₂O₂ at the anodic poles is monitored using a photomultiplier tube (PMT) or smartphone, and the images are analyzed using ImageJ software. This process triggers thionine reduction at the cathodic poles. Under the optimal conditions obtained by a face-centered central composite design (FCCD), the PMT-based detection of the BPE-ECL aptasensor exhibit a linear response over a wide dynamic range from 5 to 150ngmL^-1, with a detection limit of 0.01ngmL^-1. Additionally, smartphone-based detection shows a linear relationship between the ECL image gray value and the logarithmic concentration of the AFM1 target over a range of 10-200ngmL^-1, with a detection limit of 0.05ngmL^-1. Furthermore, the BPE-ECL aptasensor was successfully used to detect AFM1 in milk complex media without any serious interferences with reliable reproducibility (average relative standard deviation (RSD = 2.3%)). This smartphone-based detection opens a new horizon for bioanalysis that does not require a trained technician to operate and is a promising technology for point-of-care testing.