The chemiluminescence (CL) analysis based on label-free dual-aptasensor was developed for simultaneous detection of adenosine triphosphate (ATP) and chloramphenicol (CAP) in food. Magnetic microspheres and polystyrene microspheres used as separating and immobilizing carriers which immobilized the two different captured DNA, respectively. Then these carriers were put in the mixture of ATPs, CAPs, ATP-binding aptamers and CAP-binding aptamers to make one-pot label-free recognized interaction. The more ATP or CAP molecules binding their aptamers, the less aptamers left on the surface of carriers reducing the CL signals. The proposed aptasensor exhibited high selectivity and sensitivity for ATP and CAP with the limits of detection of 3.76 × 10 moL/L and 2.48 × 10 moL/L, respectively. Finally, this method is further validated by measuring the recovery of ATP/CAP spiked in three different food samples.
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