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Homovanillic acid (HVA) is a catecholamine metabolite that plays a vital role in the diagnosis of several neurological disorders. Thus, a simple and cost‐effective sensing strategy for HVA determination is significant. Herein, we describe the development of a turn‐off fluorescence sensor for the effective determination of HVA using bovine serum albumin stabilized nickel nanoclusters (BSA NiNCs). This biocompatible and water‐suspended fluorophore exhibited strong emission at 405 nm upon exciting at 325 nm. The fluorescence intensity of BSA NiNCs was efficiently quenched by the addition of HVA within the linear range 9.00×10−7 M–5.00×10−8 M with a limit of detection of 9.23×10−9 M. A static quenching mechanism was proposed for the sensor due to the aggregation of BSA NiNCs on adding HVA. The practical utility of the sensor was tested in artificial urine and blood serum and a good recovery using the spike‐recovery method were obtained.

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