A novel sensitive, competitive, and time-resolved luminescence sensor for the detection of ofloxacin (OFL) was developed in this study. The sensor used OFL-specific aptamer as a recognition molecule and rolling circle amplification (RCA) as a signal amplification tool. In this way, the time-resolved luminescence can not only avoid background noise from sample, but also provide robust luminescence for detection. Besides, the separation and enrichment of target veterinary drug can be conducted assisted by magnetic treatment. Under optimal conditions, the logarithmic correlation between the concentration of OFL and the luminescence intensity was found to be linear in the range of 5 × 10-5 × 10 mol L (R = 0.9988), with a detection limit (LOD) of 32.1 pmol L. Furthermore, this method was applied to the determination of OFL in chicken and pork samples, exhibiting good recovery (72.5-100%) and repeatability (RSD < 10.0%). These results confirm that this novel established method has good application potential for the detection of OFL in food samples.
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