General preparation method and application of high-entropy nano-enzyme

Abstract

The invention belongs to the technical field of functional nano materials and biomimetic catalysis, and discloses a universal preparation method and application of high-entropy nano enzyme. According to the method, various metal salts, polyvinylpyrrolidone, benzyl alcohol and formaldehyde are used as raw materials, and various high-entropy alloy nanoparticles and nanosheets are prepared under the same reaction condition through a solvothermal method. The prepared high-entropy nano-enzyme has the advantages of controllable composition, uniform morphology, high catalytic activity and the like, can simulate the activity of various natural enzymes, and is suitable for the fields of artificial cell systems, biological catalysis, sensing and the like.

Description

Technical Field

[0001] This invention relates to the fields of functional nanomaterials and biomimetic catalysis, and more specifically to a general method for preparing high-entropy nanozymes and their applications in biocatalysis, biosensing, and artificial cell systems. Background Technology

[0002] High-entropy alloy nanomaterials have shown broad application prospects in the field of nanozymes in recent years due to their unique compositional diversity, structural stability, and synergistic catalytic performance. Traditional nanozymes are mostly limited to single or bimetallic components, resulting in limited catalytic activity and stability. High-entropy nanozymes, through the synergistic effect of multiple metal elements, can simulate the catalytic activity of various natural enzymes, such as peroxidase and glucose oxidase, and have already been preliminarily applied in biosensing and disease diagnosis. For example, a detection platform constructed using PdMoPtCoNi high-entropy nanozymes, with its abundant active sites and tunable electronic structure, has achieved ultrasensitive detection of biomarkers such as glucose and sarcosine, providing a new tool for digital health monitoring. However, the preparation of high-entropy nanozymes still faces challenges such as difficulty in compositional control, uneven morphology, and stringent synthesis conditions. Currently, there is a lack of a universal, efficient, and scalable synthesis method capable of controllably preparing various high-entropy nanozymes.

[0003] In existing technologies, a "double-layer ice recrystallization" strategy can synthesize high-entropy nanoparticles containing multiple immiscible metals, but it requires sub-zero temperatures and relies on special quenching and annealing processes. This is complex and difficult to scale up. Furthermore, the construction of multi-level structures is limited by ice crystal control conditions, making it difficult to guarantee the uniformity of product morphology and composition. Another traditional method, high-temperature pyrolysis or hot-injection, either requires extreme reaction conditions (such as high-temperature calcination or inert gas protection) or is only applicable to specific metal systems. It often requires the use of substrates or end-capping ligands to prevent particle coarsening, which not only increases post-processing steps but also masks some catalytically active sites, reducing enzymatic performance. These methods fail to simultaneously achieve versatility, simplicity, and scalability, severely hindering the industrial application of high-entropy nanozymes.

[0004] Therefore, developing a high-entropy nanozyme preparation method that is versatile, easy to operate, and has tunable morphology and activity is an urgent problem to be solved by those skilled in the art. Summary of the Invention

[0005] In view of this, the present invention provides a general preparation method for high-entropy nanozymes and their applications. Using various metal salts, polyvinylpyrrolidone (PVP), benzyl alcohol and formaldehyde as raw materials, a series of high-entropy alloy nanoparticles (HEANPs) and nanosheets (HEANSs) are prepared efficiently and controllably by a solvothermal method, and their application in biomimetic catalysis is demonstrated.

[0006] The solvothermal method for benzyl alcohol proposed in this invention eliminates the need for extreme low or high temperature conditions. Through simple mixing, ultrasonic dispersion, and a mild solvothermal reaction of near-equimolar ratio metal salts with PVP, uniform doping of multiple metal components with controllable morphology can be achieved. This method overcomes the pain points of existing technologies, such as harsh synthesis conditions, narrow applicability, complex post-processing, and easy masking of active sites. Furthermore, the amount of raw materials can be flexibly adjusted within a reasonable range, further enhancing the practicality and universality of the method. Developing such a versatile, simple, and morphology- and activity-tunable high-entropy nanozyme preparation method is of great significance for promoting its application in artificial cells, biomedicine, and other fields.

[0007] To achieve the above objectives, the present invention provides a general method for preparing high-entropy nanozymes, comprising the following steps: (1) Dissolve various metal salts and polyvinylpyrrolidone (PVP) in benzyl alcohol and stir; (2) The mixed solution is subjected to ultrasonic treatment to ensure uniform dispersion of the system; (3) Transfer the uniformly dispersed solution to a polytetrafluoroethylene-lined high-pressure reactor and add formaldehyde dropwise; (4) Seal the reactor and react in an oven at 200 ℃ for 12-18 h; (5) After the reaction is complete, the product is naturally cooled to room temperature, washed repeatedly with a mixture of water and ethanol, centrifuged and dried to obtain high-entropy nanozymes.

[0008] Preferably, each of the various metal salts is 0.02-0.05 mmol, the polyvinylpyrrolidone is 200-300 mg, and the benzyl alcohol is 15-25 mL.

[0009] Preferably, the metal salts are used in an equimolar ratio.

[0010] Preferably, the metal salt includes, but is not limited to, a combination of ferric chloride, cobalt chloride, copper chloride, manganese chloride, palladium chloride, platinum chloride, iridium chloride, rhodium chloride, ruthenium chloride, and zinc chloride.

[0011] More preferably, the metal salt includes at least five of the metal salts mentioned above.

[0012] Preferably, the stirring speed is 450 rpm and the stirring time is 30 min.

[0013] Preferably, the ultrasonic treatment conditions are 300W and 30min.

[0014] Preferably, the amount of formaldehyde used is 0.7 mL.

[0015] Preferably, the high-entropy nanoenzyme has the morphology of spherical nanoparticles or two-dimensional nanosheets, and its size is adjustable in the range of 4-15 nm.

[0016] Preferably, the high-entropy nanozyme has glucose oxidase-like and peroxidase-like catalytic activities.

[0017] The application of the high-entropy nanozyme described in this invention in artificial cell systems, biosensing, or biomimetic catalysis.

[0018] The core technical mechanism of this invention stems from the synergistic regulation of raw material combination and reaction conditions, and the selection of each component and parameter is irreplaceable. Multiple metal salts (such as ferric chloride and cobalt chloride) are used as metal sources for high-entropy alloys. Strict adherence to near-equimolar ratios (e.g., 0.03 mmol each) is required to ensure uniform competition and coordination of multiple metal ions during the reaction, preventing phase separation due to excess of a single metal ion. This is fundamental to forming a high-entropy single-phase structure. Polyvinylpyrrolidone (PVP) possesses both dispersive and coordinating properties. The pyrrolidone groups in its molecular chain can form temporary coordination bonds with metal ions, limiting rapid reduction and aggregation. Simultaneously, it regulates particle growth rate through steric hindrance, which is crucial for achieving morphological uniformity. Benzyl alcohol not only provides a stable reaction environment as a solvent, but its hydroxyl groups can also form hydrogen bond networks with PVP, further optimizing the dispersion of metal ions. Furthermore, the hydrophobicity of the benzene ring structure can adjust the dielectric constant of the reaction system, promoting the selective reduction of metal ions by formaldehyde. Formaldehyde, as a mild reducing agent, has a highly compatible reducing capacity with the dielectric environment of the benzyl alcohol system, allowing for reduction up to 200... The reducing group is slowly released at ℃ to avoid rapid precipitation of metal ions and ensure that multiple metal ions are reduced simultaneously to form an alloy structure. If other reducing agents (such as sodium borohydride or hydrazine hydrate) are used instead, the metal components will separate due to the excessively fast reduction rate, and a high-entropy structure cannot be formed. Controlling the reaction conditions is equally essential. The combination of 30 minutes of stirring and 30 minutes of sonication is a necessary step to achieve uniform dispersion of the system. Stirring promotes the full dissolution and coordination of metal salt and PVP, while sonication breaks down the small agglomerates formed in the early stage, ensuring that metal ions are evenly distributed in the solvent, laying the foundation for subsequent simultaneous reduction. The reaction temperature of 200 ℃ and the reaction time of 12–18 hours are matched. 200 ℃ can activate the reduction activity of formaldehyde while ensuring the stability of the benzyl alcohol system and avoiding solvent decomposition or PVP failure due to high temperature. The reaction time of 12–18 hours provides sufficient time for the diffusion, coordination, reduction and alloying of metal ions. If the temperature is below 180 ℃, the reduction reaction will be incomplete and a complete alloy structure cannot be formed. If the temperature is above 220 ℃, it will lead to excessive particle growth and uncontrolled morphology. The amount of 0.7 mL of formaldehyde is precisely matched with the total molar number of metal salts to ensure complete reduction of metal ions, while avoiding side reactions (such as carbonization) caused by excessive formaldehyde, which would affect the purity of the product.

[0019] As can be seen from the above technical solution, compared with the prior art, the beneficial effects achieved by the present invention are as follows: This invention offers significant technical advantages and excellent preparation performance, enabling the universal preparation of high-entropy nanozymes. By flexibly adjusting the metal salt combination, it efficiently synthesizes high-entropy alloy nanoparticles (HEANPs) and two-dimensional nanosheets (HEANSs) with different compositions. The products exhibit uniform morphology, controllable size (4–15 nm), and absence of obvious impurity phases, demonstrating high purity in the high-entropy single-phase structure. The preparation process requires no extreme conditions (such as low-temperature quenching or high-temperature calcination), is simple to operate, has a short process, and allows for fine-tuning of raw material dosage within a reasonable range, making it suitable for large-scale production. The catalytic activity is outstanding. Leveraging the compositional diversity and synergistic effect of the high-entropy structure, the products possess multiple biomimetic enzyme catalytic activities (such as peroxidase and oxidase), with abundant and fully exposed active sites, resulting in significantly higher catalytic efficiency than traditional single-metal or bimetallic nanozymes. Simultaneously, the structural stability of the high-entropy alloy endows the products with excellent catalytic durability, maintaining high catalytic activity even after multiple cycles, thus solving the problems of limited catalytic performance and insufficient stability of traditional nanozymes. With broad application prospects, the product has important application value in fields such as biomimetic catalysis, biosensing, disease diagnosis, and environmental governance. For example, it can be used as a highly efficient catalytic probe for ultrasensitive detection of biomarkers (such as glucose and hydrogen peroxide), or as a catalytic material for pollutant degradation. Moreover, the preparation process is green and environmentally friendly, and the product has good biocompatibility, laying the foundation for the industrial application of high-entropy nanozymes. Attached Figure Description

[0020] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.

[0021] Figure 1 These are TEM images of the FeCoCuMnPd high-entropy alloy nanoparticles (HEANPs) prepared in Example 1 of this invention. Figures 1a and 1c are images under high magnification, clearly showing the regular morphology and smooth surface of the spherical nanoparticles. Figures 1b and 1d are images under low magnification, showing the overall uniform distribution of the nanoparticles.

[0022] Figure 2 Figure 2a shows the TEM images of FeCoCuMnPt HEANPs prepared in Example 2 of this invention. Figure 2b shows the image under high magnification, displaying the uniform size and crystal morphology of individual nanoparticles. Figure 2c shows the image under low magnification, displaying the overall dispersion and porous agglomeration structure of the nanoparticles. Figure 2c shows the image under medium magnification, revealing the porous interconnected features inside the agglomerates.

[0023] Figure 3 Figure 3a shows the TEM images of FeCoCuMnIr HEANPs prepared in Example 3 of this invention. Figure 3a is a high-powered image, clearly showing the morphological details of individual particles. Figure 3b is a medium-powered image (150 k×), showing the close arrangement of particles in the aggregation region. Figure 3c is a low-powered image, showing the overall distribution and aggregation morphology of the nanoparticles.

[0024] Figure 4 These are TEM images of FeCoCuMnRh HEANPs prepared in Example 4 of this invention, where images abcde are images under different magnifications, respectively. Figure 4 Figure 4a shows the overall distribution of polydisperse spherical and planar nanoparticles; Figure 4b shows the coexistence of spherical particles and planar structures; Figure 4c shows the regularity of spherical particles; Figure 4d shows the ultrathin characteristics of planar structures; Figure 4e is a high-resolution TEM (HRTEM) image showing the lattice fringes and crystal structure of the material.

[0025] Figure 5 Figure 5a shows the TEM images of FeCoCuMnRu HEANPs prepared in Example 5 of this invention. Figure 5a is a low-power image showing the overall distribution and interconnection and aggregation trend of quasi-spherical nanoparticles. Figure 5b is a medium-power image showing the structural features of interconnected aggregates. Figure 5c is a high-power image clearly showing the morphology and surface state of individual quasi-spherical particles.

[0026] Figure 6 Figure 6a shows the TEM images of the FeCoCuZnMnPd high-entropy alloy nanosheets (HEANSs) prepared in Example 6 of this invention. Figure 6a is a low-magnification image showing the overall distribution of the ultrathin two-dimensional sheet structure; Figure 6b is a medium-magnification image showing the spread-out state and size uniformity of the sheet structure; Figure 6c is a high-magnification image showing the ultrathin characteristics and edge morphology of the sheets; and Figure 6d is an HRTEM image showing the lattice structure and crystal orientation of the nanosheets. Detailed Implementation

[0027] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0028] The specific technical solution of this invention is as follows: A general preparation method for high-entropy nanozymes, comprising the following steps: Step 1: Dissolve 0.03 mmol of each metal salt and 250 mg of PVP together in 20 mL of benzyl alcohol and stir for 30 minutes; Step 2: Sonicate the mixed solution for 30 minutes to ensure uniform dispersion of the system; Step 3: Transfer the well-dispersed solution to a 100 mL polytetrafluoroethylene-lined high-pressure reactor and add 0.7 mL of formaldehyde dropwise. Step 4: Seal the reactor and place it in a 200 ℃ oven for 12–18 hours; Step 5: After the reaction is complete, allow the mixture to cool naturally to room temperature. Wash the product repeatedly with a mixture of water and ethanol, centrifuge, and then dry to obtain the high-entropy nanozyme.

[0029] The present invention will be further described in detail below with reference to embodiments: Preparation of artificial cell systems based on microfluidic technology: Example 1: Preparation of FeCoCuMnPd high-entropy alloy nanoparticles (HEANPs) Step 1: Add ferric chloride (FeCl3) 6H2O), cobalt chloride (CoCl2) 6H2O), copper chloride (CuCl2) 2H2O), manganese chloride (MnCl2) 0.03 mmol each of 4H2O and palladium chloride (PdCl2) and 250 mg PVP were added to 20 mL of benzyl alcohol and stirred at 450 rpm for 30 minutes. Step 2: 300W ultrasonic treatment for 30 minutes; Step 3: Transfer to a 100 mL high-pressure reactor and add 0.7 mL of formaldehyde dropwise; Step 4: After sealing, react at 200 °C for 12 hours; Step 5: After cooling, wash with a water / ethanol mixture, centrifuge and dry to obtain FeCoCuMnPd HEANPs.

[0030] The morphology of the obtained product is as follows Figure 1 As shown.

[0031] Example 2: Preparation of FeCoCuMnPt HEANPs The steps are the same as in Example 1, except that the metal salts are replaced with 0.03 mmol each of ferric chloride, cobalt chloride, copper chloride, manganese chloride, and platinum chloride.

[0032] The reaction time has been extended to 18 hours.

[0033] The morphology of the obtained product is as follows Figure 2 As shown.

[0034] Example 3: Preparation of FeCoCuMnIr HEANPs The steps are the same as in Example 1, except that the metal salts are replaced with 0.03 mmol each of ferric chloride, cobalt chloride, copper chloride, manganese chloride, and iridium chloride.

[0035] The reaction time is 15 hours.

[0036] The morphology of the obtained product is as follows Figure 3 As shown.

[0037] Example 4: Preparation of FeCoCuMnRh HEANPs The steps are the same as in Example 1, except that the metal salts are replaced with 0.03 mmol each of ferric chloride, cobalt chloride, copper chloride, manganese chloride, and rhodium chloride.

[0038] The reaction time is 12 hours.

[0039] The morphology of the obtained product is as follows Figure 4 As shown.

[0040] Example 5: Preparation of FeCoCuMnRu HEANPs The steps are the same as in Example 1, except that the metal salts are replaced with 0.03 mmol each of ferric chloride, cobalt chloride, copper chloride, manganese chloride, and ruthenium chloride.

[0041] The reaction time is 18 hours.

[0042] The morphology of the obtained product is as follows Figure 5 As shown.

[0043] Example 6: Preparation of FeCoCuZnMnPd high-entropy alloy nanosheets (HEANSs) Step 1: Add 0.03 mmol each of ferric chloride, cobalt chloride, copper chloride, zinc chloride, manganese chloride, and palladium chloride, along with 250 mg of PVP, to 20 mL of benzyl alcohol and stir at 450 rpm for 30 minutes. Step 2: 300W ultrasound for 30 minutes; Step 3: Transfer to a high-pressure reactor and add 0.7 mL of formaldehyde; Step 4: React at 200 °C for 12 hours to induce the formation of a two-dimensional structure using the solvent template effect; Step 5: After cooling, wash, centrifuge, and dry to obtain ultrathin HEANSs.

[0044] The morphology of the obtained product is as follows Figure 6 As shown.

[0045] To demonstrate the progressiveness of the raw material selection, dosage, and condition parameters of this invention, multiple comparative examples were set up during the research and development process, as follows: Comparative Example 1: Comparative Example with Different Solvent Substitutions Using the exact same raw materials, amounts, and reaction conditions as in Example 1, except that the solvent benzyl alcohol was replaced with an equal volume of ethanol, the product exhibited significant agglomeration after the reaction. The particle size distribution was uneven (20–50 nm), and XRD analysis revealed multiphase impurity peaks, failing to form a pure-phase high-entropy alloy structure. This is because the dielectric constant and hydrogen bonding ability of ethanol cannot form a stable dispersed network with PVP, leading to uneven metal ion reduction rates, which in turn triggers component separation and particle agglomeration, demonstrating the irreplaceable nature of benzyl alcohol as a solvent.

[0046] Comparative Example 2: Comparative Examples with Different Reducing Agent Substitutions Using the exact same raw materials, amounts, and reaction conditions as in Example 1, except that formaldehyde was replaced with an equimolar amount of sodium borohydride (NaBH4). Due to the excessively strong reducing power of sodium borohydride, metal ions precipitated rapidly, resulting in an amorphous powder with no obvious nanoparticle morphology. Furthermore, no characteristic crystal plane diffraction peaks of the high-entropy alloy could be detected, indicating that the strong reducing agent would disrupt the simultaneous reduction and alloying process of multiple metal ions, while the mild reducing properties of formaldehyde were key to the formation of the high-entropy structure.

[0047] Comparative Example 3: PVP dosage deviates from the comparative example Using the exact same raw materials, solvents, and reaction conditions as in Example 1, only the amount of PVP was adjusted to 100 mg (below the limit of this invention). After the reaction, the product particles exhibited severe agglomeration, a wide size distribution (15–35 nm), and some particles showed obvious angular defects. Catalytic activity testing showed that its peroxidase-like activity was only 32% of that of the product in Example 1. If the amount of PVP was increased to 500 mg (above the limit of this invention), excessive PVP coated the product surface, causing the catalytic active sites to be masked, and the peroxidase-like activity decreased to 45% of that of the product in Example 1. This demonstrates that the 250 mg PVP dosage is well-suited to the total molar amount of the metal salt, representing the optimal choice that balances morphological uniformity and active site exposure.

[0048] Comparative Example 4: Metal Salt Amount Not Approximately Equimolar Comparative Example Using the exact same solvent, reducing agent, PVP dosage, and reaction conditions as in Example 1, the only difference was the amount of ferric chloride adjusted to 0.1 mmol, while the other metal salts (cobalt chloride, copper chloride, manganese chloride, and palladium chloride) remained at 0.03 mmol each. XRD analysis showed that the product was dominated by an iron-based alloy phase, with other metal elements existing only as dopants. It did not form a high-entropy single-phase structure, exhibiting limited catalytic activity and poor stability. After three cycles, the activity decreased by 60%, while the product from Example 1 maintained over 85% activity after five cycles. This demonstrates that a near-equimolar ratio of multiple metal salts is the core prerequisite for forming a high-entropy structure and ensuring synergistic catalytic performance.

[0049] Comparative Example 5: Reaction temperature deviates from that of the comparative example Using the exact same raw materials and amounts as in Example 1, and other reaction conditions, only the reaction temperature was adjusted to 180 °C (below the range defined in this invention). The product after the reaction was not completely reduced, containing a large amount of metal oxide impurities, and the high-entropy alloy purity was only 65%. If the reaction temperature was adjusted to 230 °C (above the range defined in this invention), the product particles excessively grew, increasing in size to 20–40 nm, and exhibited irregular morphology. The density of catalytic active sites decreased, and the peroxidase-like activity was only 58% of that of the product in Example 1. This demonstrates that a reaction temperature of 200 °C is the optimal condition for ensuring complete reduction, alloying, and controllable morphology.

[0050] Application Example 1: Application of High-Entropy Nanozymes in Glucose Metabolism Simulation The FeCoCuMnPd high-entropy alloy nanoparticles (HEANPs) prepared in Example 1 of this invention were used as a model catalyst to investigate their bifunctional catalytic performance in glucose metabolism simulation. The specific experimental process and results are as follows.

[0051] The experiment used a 0.2 M HAc-NaAc buffer solution at pH 4.5 as the reaction system. The reaction substrates were a 10 mM glucose solution and a 0.5 mM 3,3',5,5'-tetramethylbenzidine (TMB) solution. Nanozyme concentration gradients were set at 0 μg / mL (blank group), 20 μg / mL, 40 μg / mL, 60 μg / mL, 80 μg / mL, and 100 μg / mL. After incubation at a constant temperature of 37℃ for 30 minutes, the absorbance (OD) of the reaction system at 652 nm was measured using a UV-Vis spectrophotometer. 652 The absorbance value is positively correlated with the amount of TMB oxidation products generated, and can directly reflect the strength of catalytic activity.

[0052] The results of bifunctional catalytic activity detection of FeCoCuMnPd HEANPs at different concentrations are shown in Table 1.

[0053] Table 1

[0054] OD of the blank group (without nanozyme) 652 The value was only 0.082, indicating that glucose and TMB hardly underwent spontaneous oxidation under the experimental conditions, and the catalytic reaction depended on the catalytic effect of FeCoCuMnPd HEANPs. As the nanozyme concentration increased from 20 μg / mL to 100 μg / mL, the OD of the reaction system... 652 The value linearly increased from 0.356 to 2.000, showing a clear concentration dependence, demonstrating that the catalytic activity of the nanozyme is positively correlated with its concentration, and no catalytic saturation phenomenon was observed within the tested concentration range, exhibiting good dosage control flexibility. When the nanozyme concentration was 100 μg / mL, the OD... 652 The value reached 2.000, with a relative catalytic activity 24.4 times that of the control group. This result indicates that FeCoCuMnPd HEANPs possess highly efficient bifunctional catalytic activity—first, mimicking glucose oxidase activity to oxidize glucose into gluconic acid and hydrogen peroxide (H2O2), and then mimicking peroxidase activity to catalyze the oxidation of TMB by H2O2 to generate blue oxidation products, thus completely replicating the key redox reactions in glucose metabolism. These experimental results fully validate the application potential of the high-entropy nanozyme prepared in this invention in the field of biomimetic catalysis. Its bifunctional catalytic properties can replace the combination of natural glucose oxidase and peroxidase, and it possesses advantages such as structural stability, simple preparation, and controllable cost, providing core material support for the subsequent development of glucose sensing and detection platforms and artificial metabolic systems.

[0055] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A universal method for preparing high-entropy nanozymes, characterized in that, Includes the following steps: (1) Dissolve various metal salts and polyvinylpyrrolidone in benzyl alcohol, stir, and obtain a mixed solution; (2) Ultrasonic treatment of the mixed solution; (3) Transfer the mixed solution to a high-pressure reactor and add formaldehyde; (4) After sealing, react at 200 °C for 12-18 h; (5) After the reaction is complete, the product is cooled, washed, centrifuged, and dried to obtain a high-entropy nanozyme.

2. The general preparation method of a high-entropy nanozyme according to claim 1, characterized in that, Each of the various metal salts is 0.02-0.05 mmol, the polyvinylpyrrolidone is 200-300 mg, and the benzyl alcohol is 15-25 mL.

3. The general preparation method of high-entropy nanozymes according to claim 2, characterized in that, The metal salts are used in an equimolar ratio.

4. The general preparation method of a high-entropy nanozyme according to claim 1, characterized in that, The metal salt is selected from a combination of ferric chloride, cobalt chloride, copper chloride, manganese chloride, palladium chloride, platinum chloride, iridium chloride, rhodium chloride, ruthenium chloride, and zinc chloride.

5. The general preparation method of a high-entropy nanozyme according to claim 1, characterized in that, The stirring speed was 450 rpm and the time was 30 min.

6. The general preparation method of a high-entropy nanozyme according to claim 1, characterized in that, The ultrasonic treatment conditions were 300W and 30min.

7. The general preparation method of a high-entropy nanozyme according to claim 1, characterized in that, The amount of formaldehyde used is 0.7 mL.

8. The general preparation method of a high-entropy nanozyme according to claim 1, characterized in that, The high-entropy nanozyme has the morphology of spherical nanoparticles or two-dimensional nanosheets.

9. The general preparation method of a high-entropy nanozyme according to claim 1, characterized in that, The high-entropy nanozyme exhibits glucose oxidase-like and peroxidase-like catalytic activities.

10. The application of the high-entropy nanozymes obtained by the method described in claims 1-9 in artificial cell systems, biosensing, or biomimetic catalysis.

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