Kanamycin (KAN) is widely used as a growth hormone analog and an antibacterial agent. However, abuse of this substance has resulted in the accumulation of excessive residue levels in foods of animal origin, which presents a significant risk to human health. A chemiluminescent aptasensor was constructed for the rapid quantitative detection of KAN by combining the properties of Co3O4 nanoparticles (Co3O4 NPs) nanozyme activity and DNA aptamer with high specificity. The DNA aptamer/Co3O4 NPs nanozyme regulated the chemiluminescence signal by exploiting the chemiluminescent properties of luminol oxidation by H2O2. Specific binding of KAN to the aptamer led to the formation of a steric hindrance block in the solution, which inhibited the activity of nanozyme and reduced signal intensity. The degree of signal reduction is related to the concentration of KAN. Under optimal conditions, there was good linearity between KAN concentration and chemiluminescence signal intensity in the range of 0.5-8.0 μΜ, with a detection limit of 0.26 μΜ. The detection system performed well in the presence of competing antibiotics and was virtually unaffected. The method was also suitable for the detection of KAN in milk samples with sample recoveries of 97.8%-99.1%. The chemiluminescence sensor has the advantages of low cost, specificity, and sensitivity, and does not require an external light source or modification of the nucleic acid aptamer which makes it a promising candidate for applications in the field of food detection.
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