Oxidation Of TMB Research Articles

Effective multicolor visual biosensor for ochratoxin Adetection enabled by DNAzyme catalysis and gold nanorod etching.

Anovel detection technique is introducedthat offers sensitive and reliable ochratoxin A (OTA) detection. The method leverages the etching of gold nanorods (AuNRs) stabilized by hexadecyl trimethyl ammonium bromide (CTAB) using the oxidized form of 3,3',5,5'-tetramethyl benzidine sulfate (TMB2+), creating a susceptible multicolor visual detection system for OTA.The visual detection is enabledby Mg2+-assisted DNAzyme catalysis combined with the catalytic hairpin assembly (CHA) signal amplification strategy. The presence of OTA triggers CHA and signaling responses along with the formation of G-quadruplex-hemin DNAzyme, which promotes the oxidation of TMB with H2O2, leading to the etching of AuNRs and a reduction in their aspect ratio.AuNRs experienced a blue shift in the longitudinal localized surface plasmon resonance peak, resulting in a color change.The technique has been shown to detect OTA with a low detection limit of 0.309 pg/mL, demonstrating high sensitivity and specificity. The detection technique offers versatility by enabling the detection of other pollutants through a simple replacement of the aptamer, expanding the range of detection platforms available for pollutant determinations.

Redox-mediated dsDNA-dye photooxidase mimic enable catalytic oxidation of 3,3′,5,5′-tetramethylbenzidine by dissolved O2 at neutral pH for improved biosensing

Catalytic oxidation of 3,3′,5,5′-Tetramethylbenzidine (TMB, an excellent chromogenic substrate) at neutral pH is critically important for amplified bioanalysis. Although some nanozymes exhibited the peroxidase activity at neutral pH, it is difficult to modulate their activity for homogeneous detection of biomolecules. In this work, we developed a redox-mediated dsDNA-dye photooxidase mimic that enables catalytic oxidation of TMB by dissolved O2 at neutral pH for improved biosensing. During illumination, the double-stranded DNA-SYBR Green I (dsDNA-SG) photogenerated singlet oxygen (1O2) can oxidized Mn2+ to Mn3+ that can efficiently oxidize TMB to produce a distinct blue within 4 min under neutral conditions. The catalytic oxidation of TMB can be readily modulated by the formation or dissociation of dsDNA during the sensing. After investigating a series of redox mediators, we found that only the Mn3+/Mn2+ redox mediator can lead to the oxidation of TMB at neutral pH. The maximum reaction rate of Mn2+-mediated dsDNA-SG photooxidase mimic under neutral conditions (pH 7.0) was 1.7 × 10−4 mM/s, even higher than that of horseradish peroxidase (HRP, 8.0 × 10−5 mM/s). The redox-mediated dsDNA-SG photooxidase mimic was used for detection of APE1 at pH 7.0 with over 130-fold higher sensitivity than that at 4.0, owing to the high enzymatic activity of APE1 at neutral pH. Meanwhile, we further extended this photooxidase mimic for the sensitive detection of DNA (LOD, 8 pM) and heavy metal ions at neutral pH. The redox-mediated dsDNA-dye photooxidase mimic with the ease of modulating its enzymatic activity and working at neutral pH is quite appealing for biosensing.

A Novel Approach for Colorimetric Detection of Glyphosate in Food Based on a Split Aptamer Nanostructure and DNAzyme Activity.

Glyphosate has become the most widely used herbicide worldwide in recent years. There are many concerns about toxicity and mutagenicity from long-term use of glyphosate in humans and animals. Therefore, the methods that can help in easy and quick detection of this chemical compound in food and water are critical. In this work, a biosensor was fabricated by combining the enzymatic properties of a specific DNA G-quadruplex and selectivity of a split aptamer to detect glyphosate in foods and water in a quick and simple colorimetric manner. The color change in this method is based on the oxidation of TMB by the G-quadruplex enzyme and the function of aptamer to trap glyphosate, which is visible to the naked eye in the presence and absence of the herbicide. The biosensor showed its high performance in various real samples of water and foods and provided a detection limit of 1.37 nM (R² = 0.9899) with a linear response range of 100 to 400 nM of glyphosate. This biosensor can provide an innovative, cheap and fast approach for the detection and monitoring of glyphosate in various foods and water.

Achieving ultrasensitive detection of glyphosate in fruits and vegetables using a triple-mode strategy based on carbon dots nanozymes derived from expired drugs

Rapid and sensitive detection of glyphosate (GLP) holds significant importance in the monitoring of environmental pollution and potential risks to human health. In this study, carbon dots nanozymes (CDszymes) with peroxidase-like activity were synthesized rapidly using a microwave-assisted method, employing expired drugs as raw materials. In the presence of H2O2, CDszymes catalyze the oxidation of TMB to generate blue oxTMB, which exhibits a photothermal effect under near-infrared light irradiation; an inner filter effect (IFE) may occur between oxTMB and CDszymes. By coupling the cascade catalysis of AChE and ChOx to generate H2O2, GLP effectively inhibits the activity of AChE, constructing a colorimetric/fluorescent/photothermal response platform for GLP. In colorimetry, the detection limit of GLP was 0.33 ng/mL. The detection limits of GLP by fluorescence method and photothermal method were 0.02 ng/mL and 0.41 ng/mL, respectively. The efficacy of this methodology has been successfully demonstrated in fruit and vegetable, it also provides a strategy for the high-value conversion of expired drugs.

Development of MnO2 nanosheets nanozyme-based colorimetric and photothermal dual-signal platform for monitoring alkaline phosphatase activity

Herein, a dual-signal biosensing platform for monitoring alkaline phosphatase (ALP) activity was developed. This platform combined the oxidase-mimicking activity of MnO2 nanosheets (NS) with the colorimetric and photothermal properties of oxidized 3,3′,5,5′-tetramethylbenzidine (oxTMB). MnO2 NS effectively catalyzed the oxidation of TMB to oxTMB, resulting in a color change from colorless to blue and a significant light absorption band at 652 nm. Meanwhile, oxTMB demonstrated pronounced photothermal properties, with the solution exhibiting a notable increase in temperature upon irradiation with an 808 nm laser. In the presence of ALP, 2-Phospho-L-ascorbic acid trisodium salt (AAP) was converted into L-ascorbic acid (L-AA), which then reacted with MnO2 NS. Consequently, MnO2 NS was inhibited from oxidizing TMB to oxTMB. Based on this principle, a dual-readout assay that integrated both colorimetric and photothermal signals for ALP detection was developed. The detection limits were 0.08 mU/mL for the colorimetric method and 0.12 mU/mL for the photothermal method. Furthermore, the platform was successfully applied to human serum samples, demonstrating its effectiveness in detecting ALP in complex biological matrices. The combination of colorimetric and photothermal responses enhanced both the accuracy and reliability of ALP detection, making this platform a promising tool for biomedical applications.

Near-infrared light-enhanced colorimetric signal amplification strategy for tumor marker detection based on MoS2/CuO/Au nanocomposite.

Anovel signal amplification strategy was developedby combining near-infrared light with MoS2/CuO/Au nanocomposite for building acolorimetric immunoassay. First, MoS2/CuO/Au nanocomposite was synthesized by precipitation and photoreduction methods and characterized by scanning electron microscopy (SEM) and X-ray powder diffraction (XRD). MoS2/CuO/Au nanocomposite has oxidase-like activity and can oxidize TMB to form a blue product (TMBox). Further, the catalytic oxidation of TMB was accelerated under near-infrared (NIR) laser radiation. The sandwich-type colorimetric immunoassay was constructed using MoS2/CuO/Au nanocomposite. Under the enhancement of near-infrared light, carcinoembryonic antigen (CEA) was sensitively detected in the range 0.1 to 40ng/mL with the limit of detection of 0.03ng/mL. Moreover, the immunosensor has excellent selectivity and anti-interference, good repeatability, and stability.

A Tunable Threshold Colorimetric DNA Logic Gate for Intuitive Assessment of Chemical Contaminant Exceedance.

Current molecular logic gates are predominantly focused on the qualitative assessment of target presence, which has certain limitations in scenarios requiring quantitative assessment, such as chemical contaminant monitoring. To bridge this gap, we have developed a novel DNA logic gate featuring a tunable threshold, specifically tailored to the limits of contaminants. At the core of this logic gate is a DNA-gold nanoparticle (AuNP) hybrid film that incorporates aptamer sequences to selectively bind to acetamiprid (ACE) and atrazine (ATR). Upon interaction with these contaminants, the film degrades, releasing AuNPs that, in the presence of Hg2+, catalyze the oxidation of TMB, resulting in a visible blue coloration on test paper. This aptamer-enabled process effectively establishes an OR logic gate, with ACE and ATR as inputs and the appearance of blue color as the output. A key innovation of our system is its tunable input threshold. By adjusting the concentration of Hg2+, we can fine-tune the color mutation points to match the input threshold to predefined limits, such as Maximum Residue Limits (MRLs). This alignment allows semiquantitative assessment of contaminant levels, providing intuitive visual feedback of contaminant exceedance. Validation experiments with spiked samples confirm its accuracy and reliability by closely matching HPLC results. Therefore, our colorimetric DNA logic gate is emerging as a promising tool for easy and semiquantitative monitoring of chemical contaminants across diverse applications.

A three-channel biosensor based on stimuli-responsive catalytic activity of the Fe3O4@Cu for on-site detection of tetrodotoxin in fish

Tetrodotoxin (TTX), a lethal neurotoxin, poses a grave threat to human health. The available spectroscopic methods suffer from limitations such as complex procedures and inadequate on-site capabilities. In this study, we proposed a method using Fe3O4@Cu as a catalytic biosensor combined with SERS, colorimetry and image processing for TTX detection. Integrating the aptamer amplifies the specificity of the system and masks the catalytic activity of Fe3O4@Cu. The catalytic efficiency of Fe3O4@Cu in the H2O2-TMB reaction can quantify the concentration of TTX in the system. Consequently, oxidation of TMB (oxTMB) led to the generation and change of signals for SERS, colorimetry and image processing, enabling a three-channel quantitative detection of TTX. Under the optimal conditions, the detection limit of established SERS, colorimetry and image processing were 0.055, 2.127 and 0.243 ng/mL, respectively. This three-channel biosensor was applied to real samples, providing an accurate, stable and adaptable alternative for on-site TTX detection.

Engineering an enzyme-like catalytic sensor for on-site dual-mode evaluation of total antioxidant capacity.

Anovel Fe-MoOx nanozyme, engineered with enhanced peroxidase (POD)-like activity through strategic doping and the creation of oxygen vacancies, is introducedto catalyze the oxidation of TMB with high efficiency. Furthermore, Fe-MoOx is responsive to single electron transfer (SET) and hydrogen atom transfer (HAT) mechanisms related to antioxidants and can serve as a desirable nanozyme for total antioxidant capacity (TAC) determination. The TAC colorimetric platform can reach a low LOD of 0.512μM in solution and 24.316μM in the smartphone-mediated RGB hydrogel (AA as the standard). As proof of concept, the practical application in real samples was explored. The work paves a promising avenue to design diverse nanozymes for visual on-site inspection of food quality.

A novel and facile ultraviolet-induced photo-reduction for preparing oxidase-like AuNCs@H2N-ZIF-8 composites in alcohol-water solutions

The composites of H2N-ZIF-8 supported gold nanoclusters (AuNCs@H2N-ZIF-8) were prepared by UV-induced photochemical reduction method in the methanol–water solution. The photochemical reaction was attributed to the generation of free radical as.CH2·–OH, which reduced Au(III) to Au(0) and produced monodisperse AuNCs (<2.5 nm) in the mesoporous of H2N-ZIF-8. The generated AuNCs@H2N-ZIF-8 show oxidase-like properties to catalyze the oxidation of TMB, ABTS and OPD under mild conditions. Besides, the mechanism study by EPR shows that the composites can promote oxygen to generate singlet oxygen (1O2) and superoxide radicals (·O2-), endowing it excellent catalytic activity of oxidation. Further studies revealed that the large number of exposed small AuNCs in the mesoporous pores of H2N-ZIF-8 and the highly active 211 crystalline facets formed at the edges contribute to the ROS generation intrinsically. Therefore, the present study demonstrates the significance of photochemical reduction method to prepare highly efficient oxidase-like materials, opening up a new avenue to construct noble metal materials as ultra-small nanozyme.

Bimetallic MOF-derived three-dimensional nanoflowers PdCoOx as peroxidase mimic activity for determining total antioxidant capacity

Bimetallic MOF derivatives have shown excellent performance as nano-enzymes in the field of catalysis. Herein, PdCo oxide nanoflowers with three-dimensional flower were prepared by a simple pyrolysis method on a precursor of bimetallic PdCo-MOF. PdCoOx showed excellent peroxidase mimic activity, which could significantly promote the oxidation of TMB by H2O2. Compared with CoOx, the peroxidase mimic activity of the optimized PdCoOx-300 increased by 2.41-fold. PdCoOx-300 has high affinity for TMB and H2O2 with Km values of 0.16 mM and 2.11 mM, which are only 57.03% and 36.87% of HRP, respectively. The highly specific peroxidase mimic activity is conducive to the sensitive detection of H2O2, glucose and ascorbic acid with limit of detection of 10, 100 and 10 nM, respectively. Furthermore, the total antioxidant capacity in the actual beverage samples was conducted, which showed good anti-interference ability and recovery rate.

High-performance colorimetric sensor based on PtRu bimetallic nanozyme for xanthine analysis

The identification and quantification of xanthine are crucial for assessing the freshness and quality of food products, particularly in the seafood industry. Herein, a new approach was developed, involving the in-situ controllable growth of Pt91Ru9 nanoparticles on graphitic carbon nitride to yield Pt91Ru9@C3N4 catalytic materials. By integrating Pt91Ru9@C3N4 with the xanthine/xanthine oxidase (XOD) enzyme catalytic system, a nanozyme-enzyme tandem platform was obtained for the quantification analysis of xanthine. Under the catalytic oxidation of xanthine by XOD in the presence O2, H2O2 was generated. Upon the addition of peroxidase-like activity of Pt91Ru9@C3N4, H2O2 can be decomposed into •OH and 1O2, which can further catalyze the oxidation of TMB to its oxidation product oxTMB with an absorption peak at 652 nm. This smartphone-assisted portable colorimetric sensor for visual monitoring xanthine with a low detection limit of 8.92 nmol L−1, and successfully applied to detect xanthine in grass carp and serum samples.

Multifunctional magnetic tags with photocatalytic and enzyme-mimicking properties for constructing a sensitive dual-readout ELISA

ELISA has become the gold standard for detecting harmful substances due to its specific antibody recognition and sensitive enzyme-catalyzed reactions. In this study, multifunctional magnetic Prussian blue nanolabels (MPBNs) were synthesized using a simple gentle two-step method to achieve a dual-readout mode. The MPBNs provide a sensitive colorimetric signal by efficiently catalyzing the oxidation of TMB and exhibit prominent photocatalytic degradation activity towards Rhodamine B (RhB). Supplemented by the quenching effect of oxTMB, the fluorescence was enabled to serve as a sensitive second signal. The magnetic property of the labels facilitates the separation and enrichment of the target, thereby improving sensitivity. Utilizing the versatile MPBNs, the visual limit of detection (vLOD) for Staphylococcus aureus is as low as 100 CFU/mL, with a quantitative analysis range of 102–108 CFU/mL. The introduction of photocatalytic reactions into immunoassay has opened up a new signal response system with strong momentum for development and application.

The construction of peroxidase-like Cu-BDC@FeMo6 for colorimetric detection of H2O2 and dopamine

Artificial mimic enzymes have attracted wide attention for their superior characteristics to natural enzymes in extensive applications of diagnosis and therapy. In this paper, a nanozyme Cu-BDC@FeMo6 was designed and fabricated by embedding polyoxometalate (NH4)3[FeMo6O18(OH)6]·6H2O (FeMo6) into a copper-based metal–organic framework (Cu-BDC). The integration of FeMo6 and Cu MOF based on the synergies of oxidability of FeMo6 and oxygen-driven reversible Cu+/Cu2+ enhanced the peroxidase mimicking activity, which accomplished the sensitive visual monitoring of H2O2 and dopamine (DA). The detection of H2O2 was driven by Cu-BDC@FeMo6 catalyzing the oxidation of TMB with colorimetric evolution to blue, and consecutive monitoring DA via the reverse process of reduction of ox-TMB was further achieved. The limit of detection (LOD) of H2O2 and DA were 10 μM and 2.27 μM, with excellent stability, and outstanding selectivity. The mechanism of the catalysis was further evaluated, and the generation of O2·- played a crucial role in the catalysis for oxidation. The logic gate design was constructed to illustrate the process and application. This work provides a feasible reference for the reasonable design of simulated enzyme in biosensor applications.

Colorimetric array sensor based on bimetallic nitrogen-doped carbon-based nanozyme material to detect multiple antioxidants.

Copper-cobalt bimetallic nitrogen-doped carbon-based nanoenzymatic materials (CuCo@NC) were synthesized using a one-step pyrolysis process. A three-channel colorimetric sensor array was constructed for the detection of seven antioxidants, including cysteine (Cys), uric acid (UA), tea polyphenols (TP), lysine (Lys), ascorbic acid (AA), glutathione (GSH), and dopamine (DA). CuCo@NC with peroxidase activity was used to catalyze the oxidation of TMB by H2O2 at three different ratios of metal sites. The ability of various antioxidants to reduce the oxidation products of TMB (ox TMB) varied, leading to distinct absorbance changes. Linear discriminant analysis (LDA) results showed that the sensor array was capable of detecting seven antioxidants in buffer and serum samples. It could successfully discriminate antioxidants with a minimum concentration of 10nM. Thus, multifunctional sensor arrays based on CuCo@NC bimetallic nanoenzymes not only offer a promising strategy for identifying various antioxidants but also expand their applications in medical diagnostics and environmental analysis of food.

Reactive oxygen species independent oxidase like nanozyme for dual-mode analysis of α-glucosidase

Designing highly sensitive and selective assay for analyzing α-glucosidase activity is of critical value for diagnosis of related diseases. Herein, we proposed a biomineralization synthetic route to construct bovine serum albumin templated hybrid nanomaterials of Au nanoclusters (Au NCs) and oxygen deficient manganese dioxide (indicated as BAM). BAM exhibits ultra-fast oxidase (OXD)-like activity that directly oxidized TMB to generate product. Unexpected, the OXD-like activity of BAM is not relied on the reactive oxygen species or dissolved molecular oxygen, which has rarely been reported. The superior OXD-like activity of BAM is derived from the strong interactions between Au NCs and MnO2-x nanosheets that contributes to the partial reduction of MnO2-x and increase of Au(Ⅰ) species. And the catalytic mechanism is related to electron transfer from TMB to MnO2-x with Au NCs as intermediates. The analysis of α-glucosidase activity has been realized by BAM and 2-O-a-D-glucopyranosyl-L-ascorbic acid (AA2G) as the catalytic substrate with dual mode. The AA2G can be hydrolyzed by α-glucosidase into AA, which can obviously inhibit the oxidation of TMB and recover the fluorescence of Au NCs. And the inhibition ratio and recovery efficacy are related the activity of α-glucosidase. Hence, the activity of α-glucosidase can be analyzed by the changes of the signal of absorbance or fluorescence. The dual-mode detection can prevent false negative/ positive results, satisfying different demands in various analytical situation.

Boron-doped g-C3N4 supporting Cu nanozyme for colorimetric-fluorescent-smartphone detection of α-glucosidase

BackgroundDue to that the higher activity of nanozymes would bring outstanding performance for the nanozyme-based biosensing strategies, great efforts have been made by researchers to improve the catalytic activity of nanozymes, and novel nanozymes with high catalytic activity are desired. Considering the crucial role in controlling blood glucose level, strategies like colorimetric and chemiluminescence to monitor α-glucosidase are developed. However, multi-mode detection with higher sensitivity was insufficient. Therefore, developing triple-mode detection method for α-glucosidase based on great performance nanozyme is of great importance. ResultsIn this work, a novel nanozyme Cu–BCN was synthesized by loading Cu on boron doped carbon substrate g-C3N4 and applied to the colorimetric-fluorescent-smartphone triple-mode detection of α-glucosidase. In the presence of H2O2, Cu–BCN catalyzed the generation of 1O2 from H2O2, 1O2 subsequently oxidized TMB to blue colored oxTMB. In the presence of hydroquinone (HQ), the ROS produced from H2O2 was consumed, inhibiting the oxidation of TMB, which endows the possibility of colorimetric and visual on-site detection of HQ. Further, due to that the fluorescence of Mg-CQDs at 444 nm could be quenched by oxTMB, HQ could also be quantified through fluorescent mode. Since α-glucosidase could efficiently hydrolyze α-arbutin into HQ, the sensitive detection of α-glucosidase was realized. Further, colorimetric paper-based device (c-PAD) was fabricated for on-site α-glucosidase detection. The LODs for α-glucosidase via three modes were 2.20, 1.62 and 2.83 U/L respectively, high sensitivities were realized. SignificanceThe nanozyme Cu–BCN possesses higher peroxidase-like activity by doping boron to the substrate than non-doped Cu–CN. The proposed triple-mode detection of α-glucosidase is more sensitive than most previous reports, and is reliable when applied to practical sample. Further, the smartphone-based colorimetric paper-based analytical device (c-PAD) made of simple materials could also detect α-glucosidase sensitively. The smartphone-based on-site detection provided a convenient, instrument-free and sensitive sensing method for α-glucosidase.

Cu-BTC Derived Mesoporous CuS Nanomaterial as Nanozyme for Colorimetric Detection of Glutathione.

In this paper, Cu-BTC derived mesoporous CuS nanomaterial (m-CuS) was synthesized via a two-step process involving carbonization and sulfidation of Cu-BTC for colorimetric glutathione detection. The Cu-BTC was constructed by 1,3,5-benzenetri-carboxylic acid (H3BTC) and Cu2+ ions. The obtained m-CuS showed a large specific surface area (55.751 m2/g), pore volume (0.153 cm3/g), and pore diameter (15.380 nm). In addition, the synthesized m-CuS exhibited high peroxidase-like activity and could catalyze oxidation of the colorless substrate 3,3',5,5'-tetramethylbenzidine to a blue product. Peroxidase-like activity mechanism studies using terephthalic acid as a fluorescent probe proved that m-CuS assists H2O2 decomposition to reactive oxygen species, which are responsible for TMB oxidation. However, the catalytic activity of m-CuS for the oxidation of TMB by H2O2 could be potently inhibited in the presence of glutathione. Based on this phenomenon, the colorimetric detection of glutathione was demonstrated with good selectivity and high sensitivity. The linear range was 1-20 μM and 20-300 μM with a detection limit of 0.1 μM. The m-CuS showing good stability and robust peroxidase catalytic activity was applied for the detection of glutathione in human urine samples.

A photothermal colorimetric dual-signal sensor for Staphylococcus aureus in food based on CuS nanoparticles

Staphylococcus aureus (S. aureus) is a common foodborne pathogenic organism, and rapid detection with high sensitivity and targeted killing are essential for controlling its spread. Therefore, a sandwich structure of magnetic bead-target bacteria-copper sulfide was constructed in this paper. The photothermal conversion function and peroxidase (POD) activity of CuS NPs were utilized to establish a dual sensing platform for temperature and color rendering, that is, after capturing target bacteria, light was converted to heat signal by infrared irradiation. Moreover, the specific detection of S. aureus was realized by detecting the temperature change of the complex and the oxidation of TMB by nano-enzymes of residual CuS in the supernatant to form oxTMB blue product, which formed the change of UV (ultraviolet) − Visible spectroscopy. Based on the high specificity binding and strong signal amplification of the method, the linear range of the photothermal specific probe combined with MBs-S. aureus-CuS was 10–106 CFU/mL, and the detection limit was 1.407 CFU/mL. The linear range of the color specific probe was 10–106 CFU/mL, R2 = 0.9853, and the detection limit was 1.15 CFU/mL. The photothermal efficiency of MBs-S. aureus-CuS system can reach 62.84 %, which has excellent photothermal conversion effect, and the temperature rises to about 70 °C within 7 min, which can directly kill bacteria in the detection. In addition, the portable infrared thermal imager is applied to realize the real-time field detection of S. aureus. The study was significant, because exploratory experiments were conducted to conceive a proof-of-concept based on multiple bacterial sensors that could easily be adapted to a variety of fields where bacteria pose a threat.

Core-shell structure N-doped graphene quantum dots Fe3O4/Co3O4 nanoparticles for colorimetric detection of H2O2

Hydrogen peroxide (H2O2) is an environmentally friendly intermediate in the environmental field and also plays an important role in human metabolism, but the production and accumulation of excessive H2O2 will cause harm to the human body, resulting in serious damage to cells. Therefore, N-doped graphene quantum dots Fe3O4/Co3O4 nanoparticles with core-shell structure were prepared by in-situ synthesis and precipitation. The experimental results show that the prepared Co3O4 nanoparticles have more efficient peroxidase activity than Co3O4 nanoparticles alone. It can catalyze the oxidation of 3, 3, 5, 5-tetramethylbenzidine (TMB) to produce a blue product (TMBox) in the presence of H2O2. In addition, Co3O4 with negative charge in the shell can produce a strong electrostatic driving coordination effect with positively charged TMB, which enhances the affinity between the nanoparticles and the substrate, and selectively catalyzes H2O2 oxidation of colorless TMB to turn it blue. Based on these experimental results, a simple, low-cost, and efficient colorimetric method for H2O2 detection was established, which showed a sensitive response to H2O2 in the range of 0.02–2.5 μM with a detection limit of 2.3 nM. In addition, due to the magnetic properties of the prepared nanoparticles, they could be easily recovered by applying a magnetic field. This research provides a viable approach for magnetic nanomaterials with encouraging prospects for environmental monitoring, biosensing, and more.