Surface engineering of nanozymes has been recognized as a potent strategy to improve their catalytic activity and specificity. We synthesized polydopamine-coated Co₃O₄ nanoparticles (PDA@Co₃O₄ NPs) through simple dopamine-induced self-assembly and demonstrated that these NPs exhibit catalase-like activity by decomposing H₂O₂ into oxygen and water. The activity of PDA@Co₃O₄ NPs was approximately fourfold higher than that of Co₃O₄ NPs without PDA, possibly due to the additional radical scavenging activity of the PDA shell. In addition, PDA@Co₃O₄ NPs were more stable than natural catalase under a wide range of pH, temperature, and storage time conditions. Upon the addition of a sample containing sulfide ion, the activity of PDA@Co₃O₄ NPs was significantly inhibited, possibly because of increased mass transfer limitations via the absorption of the sulfide ion on the PDA@Co₃O₄ NP surface, along with NP aggregation which reduced their surface area. The reduced catalase-like activity was used to determine the levels of sulfide ion by measuring the increased fluorescence of the oxidized terephthalic acid, generated from the added H₂O₂. Using this strategy, the target sulfide ion was sensitively determined to a lower limit of 4.3 µM and dynamic linear range of up to 200 µM. The fluorescence-based sulfide ion assay based on PDA@Co₃O₄ NPs was highly precise when applied to real tap water samples, validating its potential for conveniently monitoring toxic elements in the environment.

Paper-based test strip consisting of cerium oxide nanoparticles (nanoceria) as hydrogen peroxide (H2O2)-dependent color-changing nanozymes and cholesterol oxidase (ChOx) has been developed for convenient colorimetric determination of cholesterol without the need for chromogenic substrate. The construction of the cholesterol strip begins with physical adsorption of nanoceria on the paper surface, followed by covalent immobilization of ChOx via silanization, chitosan-mediated activation, and glutaraldehyde treatment of the nanoceria-embedded paper matrices. In the presence of cholesterol, ChOx catalyzes its oxidation to produce H2O2, which forms peroxide complex on the nanoceria surface and induces visual color change of the nanoceria-embedded paper from white/light yellow into intense yellow/orange, which was conveniently quantified with an image acquired by a conventional smartphone with the ImageJ software. Using this strategy, target cholesterol was specifically determined down to 40 μM with a dynamic linear concentration range of 0.1 to 1.5 mM under neutral pH condition, which is suitable to measure the serum cholesterol, with excellent stability during 20 days and reusability by recovering its original color-changing activity for 4 consecutive cycles. Furthermore, the practical utility of this strategy was successfully demonstrated by reliably determining cholesterol in human blood serum samples. This study demonstrates the potential of self color-changing nanozymes for developing colorimetric paper strip sensor, which is particularly useful in instrumentation-free point-of-care environments.

A reagent-free colorimetric method for galactose quantification using a composite of cerium oxide nanoparticles (nanoceria) and galactose oxidase (Gal Ox) entrapped in an agarose gel was developed. In the presence of galactose, the Gal Ox entrapped within the agarose gel catalyzed the oxidation of galactose to generate H₂O₂, which induced a color change from white to intense yellow. This reaction occurred without any chromogenic substrate. This color transition is presumed to be due to the H₂O₂-mediated alteration of the oxidation state of cerium ions present on the surface of the nanoceria. The intensity of color change was quantified by acquiring an image with a conventional smartphone, converting the image to cyan-magenta-yellow-black (CMYK) mode, and subsequently analyzing the image using the ImageJ software. Using this strategy, galactose concentration was specifically determined with excellent sensitivity of as low as 0.05 mM. The analytical utility of the assay was successfully verified by correctly determining diverse levels of galactose in human serum, which is enough to diagnose galactosemia, a genetic disorder characterized by the malfunctioning of enzymes responsible for galactose metabolism. The assay employing a hydrogel composite with entrapped nanoceria and Gal Ox, is a simple, cost-effective, and rapid colorimetric assay for galactose quantification, without using any chromogenic reagent. This cost-effective method has great potential for the diagnosis of galactosemia and is highly promising in comparison to the laborious instrumentation-based methods currently in use.

This study presents the development of luminol and gold nanoparticle co-functionalized graphene oxide (GO-AuNPs-L) hybrids as enhanced luminogenic signaling molecules in the chemiluminescence immunoassay (CLIA) for detecting C-reactive protein (CRP), a key biomarker of inflammation and cardiovascular diseases. When compared to free luminol, the GO-AuNPs-L hybrids significantly increased and prolonged the CL signal based on their synergistic enhancement in electron transfer during CL production. Based on the performance, the hybrids were employed as signaling molecules in both well plate-based and lateral flow CLIA platforms, showing substantial improvements in signal intensity and sensitivity in CRP detection. These results highlight the potential of GO-AuNPs-L hybrids as versatile and highly sensitive luminogenic molecules for immunological CRP detection, offering promising applications in clinical laboratory settings as well as in point-of-care diagnostics.

Light-activated nanozymes possess several advantages, such as light-mediated activity regulation, utilization of molecular oxygen as a green oxidant, and highly enhanced activity; however, the types of light-activated nanozymes are still limited. In this study, we found that Mg aminoclay-based Fe₃O₄/TiO₂ hybrids (MgAC-Fe₃O₄/TiO₂) exhibited peroxidase-like catalytic activity to catalyze the oxidation of the peroxidase substrate 2,2′-azino-bis(3-ethylbenzo-thiazoline-6-sulfonic acid) diammonium salt (ABTS) in the presence of H₂O₂, which was significantly enhanced under ultraviolet (UV)-light irradiation. Compared with MgAC-Fe₃O₄ and MgAC-TiO₂, MgAC-Fe₃O₄/TiO₂ showed around three-fold enhancement in the absorption intensity corresponding to the oxidized ABTS under UV-light irradiation, presumably due to the synergistic effect between Fe₃O₄ and TiO₂, thereby facilitating photocatalytic electron transfer during the catalytic action. In addition, the MgAC-Fe₃O₄/TiO₂ showed vivid stability enhancement in wide range of pH and temperature values compared with natural peroxidase. The UV-light-driven MgAC-Fe₃O₄/TiO₂-based system was successfully applied for the colorimetric detection of phenolic compounds, including pyrocatechol and resorcinol, in a dynamic linear range of 0.15–1.30 mg/mL with a limit of detection as low as 0.1 mg/mL. Further, the system could successfully determine the phenolic compounds in spiked tap water, and thus, it can be used for practical applications. We believe that the UV-light-driven enhancement in the peroxidase-like catalytic performances highlights the potential of MgAC-Fe₃O₄/TiO₂ for detecting phenolic compounds as well as other clinically and environmentally important substances.

We present a dual-mode optical sensing strategy for selective and sensitive detection of sulfide ions (S²⁻), employing copper-anchored nitrogen-doped graphene quantum dots (Cu@N-GQDs) as bifunctional nanozymes. The Cu@N-GQDs were synthesized via citric acid pyrolysis in the presence of ammonium hydroxide (serving as both nitrogen source and reductant) and copper chloride, leading to uniform incorporation of copper oxide species onto the N-GQD surface. The resulting nanohybrids exhibit two synergistic functionalities: intrinsic fluorescence comparable to pristine N-GQDs, and significantly enhanced peroxidase-like catalytic activity attributed to the anchored copper species. Upon interaction with sulfide ions, the system undergoes a dual-optical response: (i) fluorescence quenching via Cu-S complexation, and (ii) inhibition of peroxidase-like activity due to the deactivation of Cu catalytic centers via the interaction with S²⁻. This dual-signal strategy enables sensitive quantification of S²⁻, achieving detection limits of 0.5 µM (fluorescence) and 3.5 µM (colorimetry). The sensor demonstrates excellent selectivity over competing substances and high reliability and precision in real tap water samples. These findings highlight the potential of Cu@N-GQDs as robust, bifunctional, and field-deployable nanozyme probes for environmental and biomedical sulfide ion monitoring.