A class of hybrid molecularly imprinted polymeric nanoparticles (nanoMIPs) comprising the in situ formation of gold nanoparticles (AuNPs) immobilised in a molecularly imprinted D-gluconate polymer has been designed with the objective of attempting the electrochemical quantification of gluconic acid (GA) in a wine setting. The imprinted polymers were synthesised in the presence of AuNP precursors in a pre-polymerisation mixture, which were confined to one another during the polymerisation of the chains. This allowed the formation of hybrid electroactive responsive imprinted nanoparticles (hybrid AuNPs@GA-nanoMIP), which exhibited enhanced electron conductivity. The morphological characterisation of the produced nanoMIPs revealed a fully decorated Au spherical surface of 200 nm in diameter. This resulted in a large active surface area distribution, as well a pronounced electrochemical peak response at the commercial screen-printed platinum electrode (SPPtE), accompanied by enhanced electron kinetics. The AuNPs@GA-nanoMIP sensor demonstrated the ability to detect a broad range of GA concentrations (0.025-5 mg/mL) with exceptional selectivity and reproducibility. The calibration curves were fitted with different isotherm models, such as the Langmuir, Freundlich and Langmuir-Freundlich functions. Moreover, the efficacy of the detection method was demonstrated by the recovery rates observed in real samples of Italian red wine. This research contributes to the development of a robust and reliable electrochemical sensor for the on-site determination of gluconic acid in food analysis.