Dielectric breakdown technique was utlised to fabricate 5–6 nm nanopores for vanillin detection in various food samples. A highly selective aptamer (Van_74) with high binding affinity towards vanillin was used as capture probe. Under optimal conditions, aptamer/vanillin complex translocation induced deeper events than the bare aptamer. As a result, the proposed nanopore aptasensor exhibits a linear range from 0.5 to 5 nM (R² = 0.972) and a low detection limit of 500 pM, which is significantly better than conventional platforms. Furthermore, our aptasensor showed excellent immunity against different interferons and was used to detect vanillin in different food samples. The food sample measurements were confirmed with an additional UV–Vis assay, the results of the two techniques were statistically evaluated and showed no statistically significant difference. Hence, this work represents a proof-of-concept involving the design and testing of aptamer/nanopore sensors for small molecules detection, which plays a critical role in food safety.