Enzyme-Assisted Solid-Phase Microextraction Coupled with a DNA Nanowalker for Dual-Amplified Detection of Chloramphenicol in Animal-Derived Food Products

Chloramphenicol (CAP), an aminoalcohol antibiotic, exerts its action on bacterial ribosomes, thereby obstructing protein synthesis. However, the use of CAP in husbandry may lead to its excessive accumulation in animal-derived food products. This presents potential risks to consumer health. This study developed a novel dual-amplification fluorescence detection method by integrating enzyme-assisted solid-phase microextraction (SPME) with a Fe₃O₄@Au NP-based DNA nanowalker for the detection of CAP in food. The combination of a quartz rod-based SPME biosensor and DNA nanowalker effectively eliminated matrix interference, enabling the conversion of CAP and enhancement of detection signals through two cyclic amplification processes. The strategy demonstrated high sensitivity with a limit of detection of 28.1 aM as well as a wide linear range from 0.1 fM to 1 nM (with <i>R</i>² > 0.99). This method also demonstrates robust stability and accuracy in detecting trace amounts of CAP in both authentic and prepared positive samples.