Nanozymes, which have high enzyme-like activity of natural enzymes, are very promising for analytical purposes, in particular, for the development of methods for sensitive, quantitative detection of practically important analytes – biomarkers of common diseases or pharmaceutical products. Recently, it has been reported that artificial enzymes with laccase-like activity or “nanolaccases (nLacs),” can serve as catalytic elements for the creation of sensitive methods for catecholamines. Our work aimed to obtain laccase-like nanozymes and characterize and demonstrate their suitability for spectrophotometric adrenaline (AD) analysis. In this article, we report on preparing five hexacyanoferrate nanoparticles (HCF NPs) that possess laccase-like activity, particularly, Co-HCF, Ni-HCF, Mn-HCF, Zn-HCF, and Cu-HCF. Among the investigated nLacs, Cu-HCF was selected and characterized. It was shown that Cu-HCF reveals the highest activities, is stable in various pH conditions in the range 3.0–6.5, and has satisfactory stored stability. A new spectrophotometric method for the quantitative detection of AD was created using the selected nLacs. The linearity of the proposed method is in the range from 5 μM to 50 μM (0.66–11 μg/ml), and the limit of detection is 1.5 μM (0.33 μg/ml), which is lower than that catalyzed by native laccase (1.15 μg/ml). The proposed method was tested on the real samples of pharmaceuticals, and the obtained data agree with the data declared by the producer. The resulting nLacs have great potential for use in catalysis of mimetics, environmental restoration, and sensor design. Thus methods, the obtained Cu-HCF has great potential application in spectrophotometric and biosensor method for analysis of biologically active toxic compounds in surface waters.