CN118165978A - Sequential nano catalyst and preparation method thereof - Google Patents

Sequential nano catalyst and preparation method thereof Download PDF

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CN118165978A
CN118165978A CN202410277726.3A CN202410277726A CN118165978A CN 118165978 A CN118165978 A CN 118165978A CN 202410277726 A CN202410277726 A CN 202410277726A CN 118165978 A CN118165978 A CN 118165978A
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graphene oxide
mesoporous organic
organic silicon
silicon oxide
pmo
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王庆
杨真露
万志敏
许梓
彭云松
唐雷
王荣品
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Guizhou Provincial Peoples Hospital
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    • C12Y101/03—Oxidoreductases acting on the CH-OH group of donors (1.1) with a oxygen as acceptor (1.1.3)
    • C12Y101/03004—Glucose oxidase (1.1.3.4)

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Abstract

The invention provides a sequential nano catalyst and a preparation method thereof, and belongs to the technical field of catalysts. The invention provides a preparation method of a sequential nano catalyst, which comprises the following steps: mixing graphene oxide coated mesoporous organic silicon oxide, a first solvent and an activating agent, and performing carboxyl activation treatment to obtain activated graphene oxide coated mesoporous organic silicon oxide; and mixing the activated graphene oxide coated mesoporous organic silicon oxide with glucose oxidase, and performing condensation reaction to obtain the sequential nano catalyst. According to the invention, firstly, carboxyl activation treatment is carried out on the graphene oxide coated mesoporous organic silicon oxide, and then glucose oxidase is connected to the surface of the mesoporous organic silicon oxide through amino-carboxyl condensation reaction, so that the glucose oxidase in the sequential nano catalyst exerts glucose oxidase activity, and the graphene oxide exerts peroxidase activity and photo-thermal property, so that a nano reagent with sequential catalysis is formed.

Description

Sequential nano catalyst and preparation method thereof
Technical Field
The invention belongs to the technical field of catalysts, and particularly relates to a sequential nano catalyst and a preparation method thereof.
Background
The enzyme has active catalytic performance and substrate specificity, and is important to ensure the functions and metabolism of cells. Most natural enzymes are proteins or RNAs, which suffer from the disadvantages of instability, high cost and complex synthesis. With the development of nanotechnology, some nanomaterials with enzyme-like activity have been developed, called "nanoenzymes". Compared with natural enzymes, the nano-enzyme has higher stability and lower cost, and can be produced in batches. Different nanoenzymes may exert different enzyme activities, such as catalase-like activity, peroxidase-like activity, oxidase-like activity, and superoxide dismutase-like activity. Nanoezymes have been studied in biomedical analysis, imaging, environmental protection, and the like.
The characteristics of the tumor microenvironment include excessive hydrogen peroxide, hypoxia, acidity, low activity of hydrogenase and the like. The nano enzyme can utilize the characteristics of tumor microenvironment to exert corresponding enzyme activity and generate cytotoxicity molecules such as Reactive Oxygen Species (ROS), including singlet oxygen (1O2), superoxide anions (O 2), hydroxyl radicals (OH) and the like, which are beneficial to killing tumor cells.
Graphene Oxide (GO) is a two-dimensional platelet nanomaterial with excellent chemical/mechanical stability, strong Near Infrared (NIR) region absorption, good thermal and electrical conductivity. It has been widely studied in the fields of nanoelectronics, biosensors, drug delivery, photothermal therapy, and bioimaging. In addition, GO also exhibits peroxidase-like activity, which can catalyze the generation of hydroxyl radicals from H 2O2. Based on this property, GO has been studied for glucose detection, but its use in tumor therapy has not been fully studied, and thus it is necessary to further explore its antitumor potential. However, there are some challenges in the use of GO for organisms, and studies have shown that GO has a concentration-dependent cytotoxicity on human cells, which is thought to be caused by the direct physical damage to cell membranes by the sharp edges of GO.
Mesoporous organic silica (PMO) is a mesoporous material with organic and inorganic frameworks. The modified mesoporous material has larger specific surface area, ordered mesopores, surface characteristics easy to modify and good biocompatibility and biodegradability. Many studies have constructed multifunctional PMO-based nanosystems that can be used for drug delivery, bioimaging, combined treatment of tumors, etc. The shape and size of the PMO can be adjusted relatively easily by optimizing the amount of surfactant, the ratio of ethanol and water, the silicon source dose, and the concentration of ammonia. Studies have reported spherical, egg yolk, double, deformable hollow and microminiature PMO, etc. Research shows that GO can be coated on the surface of spherical mesoporous silica nano particles to obtain a multifunctional nano platform which is used for tumor treatment and has good biocompatibility, but does not have sequential catalytic activity. Therefore, constructing a catalyst with sequential catalytic activity for the combined synergistic treatment of tumors becomes a technical problem to be solved in the art.
Disclosure of Invention
The invention aims to provide a sequential nano catalyst and a preparation method thereof. GOD in the sequential nano-catalyst prepared by the preparation method provided by the invention exerts glucose oxidase activity, GO exerts peroxidase activity and photo-thermal property, and a sequentially catalyzed nano-reagent is formed.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a preparation method of a sequential nano catalyst, which comprises the following steps:
(1) Mixing graphene oxide coated mesoporous organic silicon oxide, a first solvent and an activating agent, and performing carboxyl activation treatment to obtain activated graphene oxide coated mesoporous organic silicon oxide;
(2) And (3) mixing the activated graphene oxide coated mesoporous organic silicon oxide obtained in the step (1) with glucose oxidase, and performing condensation reaction to obtain the sequential nano catalyst.
Preferably, the preparation method of the graphene oxide coated mesoporous organic silicon oxide in the step (1) comprises the following steps:
1) Mixing a surfactant, a second solvent and a catalyst to obtain a mixed solution;
2) Mixing the mixed solution obtained in the step 1), a silicon source and 3-aminopropyl trimethoxy silane, and performing hydrolytic polycondensation reaction to obtain mesoporous organic silicon oxide;
3) And 2) mixing the mesoporous organic silicon oxide obtained in the step 2), water and graphene oxide solution, and performing electrostatic self-assembly to obtain the graphene oxide coated mesoporous organic silicon oxide.
Preferably, the mass ratio of mesoporous organic silica to graphene oxide in the step 3) is 2: (1-2).
Preferably, the activator in the step (1) is EDC solution and NHS solution.
Preferably, the concentration of the EDC solution and the NHS solution are independently 1-2 mg/mL.
Preferably, in the step (1), the mass ratio of the graphene oxide coated mesoporous organic silica to EDC to NHS is 1: (1-1.2): (1-1.4).
Preferably, the temperature of the carboxyl group activation treatment in the step (1) is room temperature, and the time of the carboxyl group activation treatment is 2-3 hours.
Preferably, the mass ratio of the graphene oxide coated mesoporous organic silica in the step (1) to the glucose oxidase in the step (2) is 1: (2-3).
Preferably, the temperature of the condensation reaction in the step (2) is room temperature, and the time of the condensation reaction is 20-24 hours.
The invention also provides the sequential nano catalyst prepared by the preparation method.
The invention provides a preparation method of a sequential nano catalyst, which comprises the following steps: mixing graphene oxide coated mesoporous organic silicon oxide, a first solvent and an activating agent, and performing carboxyl activation treatment to obtain activated graphene oxide coated mesoporous organic silicon oxide; and mixing the activated graphene oxide coated mesoporous organic silicon oxide with glucose oxidase, and performing condensation reaction to obtain the sequential nano catalyst. According to the invention, firstly, carboxyl activation treatment is carried out on the graphene oxide coated mesoporous organic silicon oxide, and then glucose oxidase is connected to the surface of the mesoporous organic silicon oxide through amino-carboxyl condensation reaction, so that the glucose oxidase in the sequential nano catalyst exerts glucose oxidase activity, and the graphene oxide exerts peroxidase activity and photo-thermal property, so that a nano reagent with sequential catalysis is formed. The results of the examples show that GOD exerts glucose oxidase activity and GO exerts peroxidase activity and photo-thermal properties in the sequential nano-catalyst prepared by the preparation method of the invention, and a nano-reagent with sequential catalysis is formed.
Drawings
FIG. 1 is a transmission electron microscope image of PMO in example 1;
FIG. 2 is a transmission electron microscopy image of PMO@GO of example 1;
FIG. 3 is an ultraviolet-visible absorption spectrum of PMO, PMO@GO, GOD, and PMO@GO-GOD of example 1;
FIG. 4 is a Fourier infrared spectrum of PMO@GO and PMO@GO-GOD in example 1;
FIG. 5 shows the color change of PMO@GO-GOD and PMO@GO mixed with a glucose oxidase activity detecting reagent at different concentrations;
FIG. 6 shows absorbance at 500nm after mixing PMO@GO-GOD and PMO@GO with a glucose oxidase activity detecting reagent at different concentrations;
FIG. 7 shows the color change of different groups mixed with equal amounts of H 2O2 and TMB;
FIG. 8 shows absorbance at 652nm after mixing different groups with equal amounts of H 2O2 and TMB;
FIG. 9 shows the temperature change of PMO@GO-GOD solutions of different concentrations irradiated with 808nm (1.0W/cm 2) laser light;
FIG. 10 shows the temperature change of PMO@GO-GOD solution irradiated under 808nm lasers with different powers.
Detailed Description
The invention provides a preparation method of a sequential nano catalyst, which comprises the following steps:
(1) Mixing graphene oxide coated mesoporous organic silicon oxide, a first solvent and an activating agent, and performing carboxyl activation treatment to obtain activated graphene oxide coated mesoporous organic silicon oxide;
(2) And (3) mixing the activated graphene oxide coated mesoporous organic silicon oxide obtained in the step (1) with glucose oxidase, and performing condensation reaction to obtain the sequential nano catalyst.
The source of each raw material is not particularly limited, and commercially available products known to those skilled in the art may be used.
According to the invention, graphene oxide coated mesoporous organic silicon oxide, a first solvent and an activating agent are mixed, and carboxyl activation treatment is carried out, so that activated graphene oxide coated mesoporous organic silicon oxide is obtained.
In the invention, the preparation method of the graphene oxide coated mesoporous organic silicon oxide preferably comprises the following steps:
1) Mixing a surfactant, a second solvent and a catalyst to obtain a mixed solution;
2) Mixing the mixed solution obtained in the step 1), a silicon source and 3-aminopropyl trimethoxy silane, and performing hydrolytic polycondensation reaction to obtain mesoporous organic silicon oxide;
3) And 2) mixing the mesoporous organic silicon oxide obtained in the step 2), water and graphene oxide solution, and performing electrostatic self-assembly to obtain the graphene oxide coated mesoporous organic silicon oxide.
In the present invention, the surfactant, the second solvent and the catalyst are preferably mixed to obtain a mixed solution.
In the present invention, the surfactant is preferably cetyltrimethylammonium bromide or cetyltrimethylammonium chloride.
In the present invention, the second solvent is preferably ethanol and water; the volume ratio of the ethanol to the water is preferably 0.3: (1 to 1.3), more preferably 0.3:1.3.
In the present invention, the catalyst is preferably aqueous ammonia; the mass concentration of the ammonia water is preferably 22-28%, more preferably 25%; the volume ratio of the catalyst to the second solvent is preferably 1: (150 to 160), more preferably 1:160. the catalyst can accelerate the reaction speed and reduce the reaction temperature.
In the present invention, the mixing of the surfactant, the second solvent and the catalyst is preferably performed under stirring conditions; the temperature of the stirring is preferably 30-50 ℃, more preferably 35 ℃; the stirring time is preferably 1 to 2 hours. The stirring speed is not particularly limited, so long as the raw materials are uniformly mixed.
After the mixed solution is obtained, the mixed solution, a silicon source and 3-aminopropyl trimethoxy silane are preferably mixed for hydrolysis and polycondensation reaction to obtain the mesoporous organic silicon oxide.
In the present invention, the silicon source is preferably ethyl orthosilicate and bis [3- (triethoxysilyl) propyl ] tetrasulfide; the volume ratio of the tetraethoxysilane to the bis [3- (triethoxysilyl) propyl ] tetrasulfide is preferably (8-9): 1, more preferably 9:1.
In the present invention, the volume ratio of the second solvent to the silicon source is preferably (700 to 800): 1, more preferably 800:1.
In the present invention, the volume ratio of the mass of the surfactant to the silicon source is preferably (4 to 5) g:10mL, more preferably 4g:10mL.
In the present invention, the volume ratio of the silicon source to 3-aminopropyl trimethoxysilane is preferably 20: (1-2), more preferably 20:1. the invention synthesizes mesoporous organic silicon oxide with positive charges by adopting 3-aminopropyl trimethoxy silane.
In the invention, the mixing of the mixed solution, the silicon source and the 3-aminopropyl trimethoxysilane is preferably performed by rapidly adding the silicon source into the mixed solution and then adding the 3-aminopropyl trimethoxysilane. The invention can be used for quickly adding a silicon source to uniformly nucleate.
The rapid addition operation is not particularly limited in the present invention, and may be performed by an operation well known to those skilled in the art.
In the present invention, the 3-aminopropyl trimethoxysilane is preferably added 15 minutes after the silicon source is added.
In the present invention, the temperature of the hydrolytic polycondensation reaction is preferably room temperature; the hydrolysis polycondensation reaction time is preferably 20 to 30 hours, more preferably 24 hours.
After the hydrolysis and polycondensation reaction is completed, the product obtained by the hydrolysis and polycondensation reaction is preferably subjected to primary centrifugation, washing, extraction, secondary centrifugation, repeated extraction, secondary centrifugation and drying in sequence, so that the mesoporous organic silicon oxide is obtained.
The present invention is not particularly limited to the above-described one-time centrifugation operation, and may be performed by any operation known to those skilled in the art.
In the present invention, the detergent used for the washing is preferably ethanol; the number of times of washing is preferably 3.
In the invention, the extracting agent adopted in the extraction is preferably a mixed solution of concentrated hydrochloric acid and ethanol; the volume ratio of the concentrated hydrochloric acid to the ethanol is preferably 1:500; the temperature of the extraction is preferably 60 ℃; the extraction time is preferably 3 hours. The concentration of the concentrated hydrochloric acid is not particularly limited, and may be any concentration known to those skilled in the art.
The secondary centrifugation operation is not particularly limited, and may be performed by any operation known to those skilled in the art.
In the present invention, the number of times of repeating extraction and secondary centrifugation is preferably 2. The invention can remove the surfactant in the material by adopting extraction and secondary centrifugation.
The drying operation is not particularly limited, and may be performed by any operation known to those skilled in the art.
After mesoporous organic silicon oxide is obtained, the mesoporous organic silicon oxide, water and graphene oxide solution are preferably mixed, and electrostatic self-assembly is carried out to obtain graphene oxide coated mesoporous organic silicon oxide.
In the invention, the mass ratio of the mesoporous organic silicon oxide to the graphene oxide is preferably 2: (1-2), more preferably 2:1.
The water consumption and the concentration of the graphene oxide solution are not particularly limited, so long as the mass ratio of mesoporous organic silicon oxide to graphene oxide is ensured to be 2: (1-2).
In the present invention, the mixing of the mesoporous organic silica, water and the graphene oxide solution is preferably mixing the mesoporous organic silica and water, and then mixing with the graphene oxide solution.
In the present invention, the temperature of the electrostatic self-assembly is preferably room temperature; the time of the electrostatic self-assembly is preferably 20-30 hours, more preferably 24 hours; the electrostatic self-assembly is preferably carried out under light-protected conditions. The invention can prevent raw materials from oxidation by carrying out electrostatic self-assembly under the light-shielding condition.
After the electrostatic self-assembly is completed, the product obtained by the electrostatic self-assembly is preferably subjected to centrifugation and washing in sequence, so that the graphene oxide coated mesoporous organic silicon oxide is obtained.
The operation of the centrifugation and washing is not particularly limited in the present invention, and may be performed by an operation well known to those skilled in the art.
According to the invention, APTES is added to synthesize mesoporous organic silicon oxide with positive charges, the diameter is about 70nm, and then the graphene oxide with negative charges is wrapped on the surface of PMO to obtain PMO@GO, so that the nano particles still keep regular morphology and good dispersibility.
In the present invention, the first solvent is preferably water.
In the present invention, the activator is preferably EDC solution and NHS solution; the concentration of the EDC solution and the NHS solution is independently preferably 1-2 mg/mL; the solvent of the activator is preferably water. The invention adopts the activator to activate carboxyl, which is favorable for the subsequent condensation reaction.
In the invention, the mass ratio of the graphene oxide coated mesoporous organic silicon oxide to EDC to NHS is preferably 1: (1-1.2): (1 to 1.4), more preferably 1:1.2:1.4.
The operation of mixing the graphene oxide coated mesoporous organic silicon oxide, the first solvent and the activating agent is not particularly limited, and the technical scheme for preparing the mixed material, which is well known to the person skilled in the art, can be adopted.
In the present invention, the temperature of the carboxyl group activation treatment is preferably room temperature; the time for the carboxyl group activation treatment is preferably 2 to 3 hours.
After activated graphene oxide coated mesoporous organic silicon oxide is obtained, the activated graphene oxide coated mesoporous organic silicon oxide is mixed with glucose oxidase, and condensation reaction is carried out, so that the sequential nano catalyst is obtained.
In the invention, the mass ratio of the graphene oxide coated mesoporous organic silicon oxide to the glucose oxidase is preferably 1: (2-3), more preferably 1:2.
The operation of mixing the activated graphene oxide coated mesoporous organic silicon oxide and glucose oxidase is not particularly limited, and the technical scheme for preparing the mixed material, which is well known to the person skilled in the art, can be adopted.
In the present invention, the temperature of the condensation reaction is preferably room temperature; the time of the condensation reaction is preferably 20 to 24 hours, more preferably 24 hours.
After the condensation reaction is completed, the invention preferably carries out centrifugation and washing on the product obtained by the condensation reaction in sequence to obtain the sequential nano catalyst.
The operation of the centrifugation and washing is not particularly limited in the present invention, and may be performed by an operation well known to those skilled in the art.
According to the invention, glucose Oxidase (GOD) is connected to the surface of the graphene oxide coated mesoporous organic silicon oxide through an amino-carboxyl condensation reaction, and GOD in the nano particles exerts glucose oxidase activity, GO exerts peroxidase activity and photo-thermal performance, so that a sequentially catalyzed nano reagent is formed.
According to the sequential nano catalyst provided by the invention, the two-dimensional flaky graphene oxide is wrapped on the surface of the spherical mesoporous silicon oxide, so that the cell damage caused by the sharp edge of the two-dimensional material is reduced, and animal experiments show that the nano material has good biocompatibility.
According to the invention, firstly, carboxyl activation treatment is carried out on the graphene oxide coated mesoporous organic silicon oxide, and then glucose oxidase is connected to the surface of the mesoporous organic silicon oxide through amino-carboxyl condensation reaction, so that the glucose oxidase in the sequential nano catalyst exerts glucose oxidase activity, and the graphene oxide exerts peroxidase activity and photo-thermal property, so that a nano reagent with sequential catalysis is formed.
According to the invention, two-dimensional flaky GO is wrapped on the surface of spherical PMO, and Glucose Oxidase (GOD) is loaded on the surface, so that a simple multifunctional nano system PMO@GO-GOD with good biocompatibility and sequential catalytic activity is constructed, and the multifunctional nano system PMO@GO-GOD is used for the catalytic treatment and the combined photothermal treatment of breast cancer; wherein, GO is wrapped on the surface of PMO, which can reduce the damage of the two-dimensional structure to the cell membrane and improve the biocompatibility; the loaded GOD can decompose glucose in the tumor microenvironment into H 2O2, and then GO catalyzes the conversion of H 2O2 into hydroxyl radicals through the peroxidase-like activity of the GOD so as to kill tumor cells; in addition, the GO has higher photo-thermal conversion capability, so that effective photo-thermal treatment can be realized under the irradiation of laser; the PMO@GO-GOD has good glucose oxidase and peroxidase-like activity, and can effectively generate active oxygen. The results of the cell and animal experiments show that PMO@GO-GOD can realize the combination of catalytic treatment and photo-thermal treatment, obviously increase the generation of hydroxyl free radicals and effectively kill tumors. Furthermore, PMO@GO-GOD has good biosafety in vivo.
The invention also provides the sequential nano catalyst prepared by the preparation method.
The technical solutions of the present invention will be clearly and completely described in the following in connection with the embodiments of the present invention. It will be apparent that the described embodiments are only some, but not all, embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
The preparation method of the sequential nano catalyst comprises the following steps:
(1) 0.1g of CTAB, 37.5mL of ethanol, 162.5mL of pure water and 1.25mL of concentrated ammonia water were stirred at 35℃for 1 hour to obtain a mixed solution; wherein the mass concentration of the concentrated ammonia water is 25%;
(2) Fully and uniformly mixing 0.225mLTEOS and 0.025mLTESPTS, quickly adding the mixture into the mixed solution obtained in the step (1), adding 12.5uLAPTES after 15min, continuing to react for 24h, centrifuging, and washing with ethanol for 3 times to obtain a product;
(3) Resuspending the product obtained in the step (2) in a mixed solution of concentrated hydrochloric acid and ethanol with the mass concentration of 37%, stirring for 3 hours at 60 ℃, then repeating the step after centrifugation for three times, and drying to obtain mesoporous organic silicon oxide which is marked as PMO; wherein, the volume ratio of the concentrated hydrochloric acid to the ethanol is 1:500;
(4) Mixing 5mg of mesoporous organic silicon oxide with water, then mixing with graphene oxide solution, stirring for 24 hours at room temperature in a dark place for electrostatic self-assembly, centrifuging, and washing with pure water for 3 times to obtain graphene oxide coated mesoporous organic silicon oxide, which is denoted as PMO@GO; wherein, the mass ratio of mesoporous organic silicon oxide to graphene oxide is 2:1, a step of;
(5) Mixing 1mg of the graphene oxide coated mesoporous organic silicon oxide obtained in the step (4) with 1.2mL of EDC solution and 1.4mLNHS of solution, and activating for 2 hours at room temperature to obtain activated graphene oxide coated mesoporous organic silicon oxide; wherein, the concentration of the EDC solution and the NHS solution is 1mg/mL;
(6) And (3) adding 2mg of GOD into the activated graphene oxide coated mesoporous organic silicon oxide obtained in the step (5), performing condensation reaction for 24 hours at room temperature, centrifuging, and washing with pure water for 3 times to obtain a sequential nano catalyst, which is named as PMO@GO-GOD.
FIG. 1 is a transmission electron microscope image of PMO in example 1; FIG. 2 is a transmission electron microscopy image of PMO@GO of example 1; FIG. 3 is an ultraviolet-visible absorption spectrum of PMO, PMO@GO, GOD, and PMO@GO-GOD of example 1; FIG. 4 is a Fourier infrared spectrum of PMO@GO and PMO@GO-GOD of example 1.
The core of the sequential nanocatalyst prepared in example 1 is mesoporous organic silica, PMO, which is positively charged due to the introduction of amino groups during synthesis, and as can be seen from the electron microscopy image of fig. 1, PMO is a regular spherical particle with a diameter of about 70nm, a surface area of about 1647m 2 g-1, a pore volume of about 1.57cm 3 g-1, and a pore diameter of about 3.7nm; after the graphene oxide is wrapped, the charges of the nano particles PMO@GO are converted into negative charges, and good morphology and dispersibility are maintained; after GOD is connected, the nanoparticle PMO@GO-GOD shows an absorption peak at 970nm, which indicates that GOD is successfully connected; the Fourier infrared spectrum of FIG. 4 shows that PMO@GO-GOD shows absorption peaks of amide bands 1 and 2, indicating successful GOD ligation.
1. Evaluation of glucose oxidase and peroxidase Activity of PMO@GO-GOD prepared in example 1
1. And detecting the glucose oxidase activity of the PMO@GO-GOD by using a glucose oxidase detection kit.
900UL of detection working solution +100 mu LPMO@GO-GOD (different concentrations, 0, 31.25, 62.5, 125 and 250 mu g/mL, calculated as PMO equivalent and prepared by pure water), incubating for 2 hours, and testing the absorbance of the solution at 500nm by using an enzyme-labeled instrument; 900uL of detection working solution +100 mu LPMO@GO (different concentrations, 0, 31.25, 62.5, 125 and 250 mu g/mL, calculated as PMO equivalent, prepared with pure water), incubated for 2h, and the absorbance of the solution at 500nm was measured with an enzyme-labeled instrument as a control group. Wherein each set of experiments included three secondary wells.
FIG. 5 shows the color change of PMO@GO-GOD and PMO@GO mixed with a glucose oxidase activity detecting reagent at different concentrations; FIG. 6 shows absorbance at 500nm after mixing PMO@GO-GOD and PMO@GO with a glucose oxidase activity detection reagent at different concentrations.
As can be seen from fig. 5 and 6, as the pmo@go-GOD concentration increases, the color of the solution changes from colorless to orange-red and increases gradually, with a corresponding increase in absorbance at 500 nm; the control group showed no obvious color or absorbance change, indicating that PMO@GO-GOD had glucose oxidase activity.
2. The peroxidase activity of PMO@GO-GOD was detected using Tetramethylbenzidine (TMB).
TMB, when oxidized, produces a distinct green color. The groups are four: glucose (GLU) +PMO, GLU+PMO@GO, PMO@GO-GOD, GLU+PMO@GO-GOD, concentrations were 250 μg/mL (calculated as PMO equivalent). Each group contained an equal amount of H 2O2 (final concentration 50 mM) and TMB (final concentration 800 nM). After incubation of each group for 30min, the color change was observed and the absorbance at 652nm was measured for each group of solutions.
FIG. 7 shows the color change of different groups mixed with equal amounts of H 2O2 and TMB; FIG. 8 shows absorbance at 652nm after mixing different groups with equal amounts of H 2O2 and TMB.
As can be seen from fig. 7 and 8, the glu+pmo@go group has significantly higher absorbance than the glu+pmo group, indicating that GO does exert peroxidase activity therein; the absorbance of the GLU+PMO@GO-GOD group is significantly higher than that of the GLU+PMO@GO group and the PMO@GO-GOD group, indicating that the amount of H 2O2 can be increased after GOD is connected, thereby enhancing the peroxidase activity of GO.
2. Evaluation of the photo-thermal efficacy of PMO@GO-GOD prepared in example 1
PMO@GO-GOD solutions (0, 0.675,1.25,2.5,5mg/mL, calculated as equivalent PMO, prepared with pure water) of different concentrations were irradiated with a 808nm (1.0W/cm 2) laser, and the change in solution temperature with time was recorded. PMO@GO-GOD solution (3.5 mg/mL, calculated as equivalent amount of PMO) was irradiated with 808nm laser light (0.25,0.5,1.0 and 2.0W/cm 2) at different powers, and the change in solution temperature over time was recorded.
FIG. 9 shows the temperature change of PMO@GO-GOD solutions of different concentrations irradiated with 808nm (1.0W/cm 2) laser light; FIG. 10 shows the temperature change of PMO@GO-GOD solution irradiated under 808nm lasers with different powers.
As can be seen from fig. 9 and 10, the pmo@go-GOD solution temperature increases with increasing irradiation time, and the rate and extent of increase with increasing solution concentration and laser power.
From the above examples, it can be seen that GOD in the sequential nano-catalyst prepared by the preparation method provided by the invention exerts glucose oxidase activity, GO exerts peroxidase activity and photo-thermal property, and a nano-reagent with sequential catalysis is formed.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.

Claims (10)

1. A method for preparing a sequential nanocatalyst, comprising the steps of:
(1) Mixing graphene oxide coated mesoporous organic silicon oxide, a first solvent and an activating agent, and performing carboxyl activation treatment to obtain activated graphene oxide coated mesoporous organic silicon oxide;
(2) And (3) mixing the activated graphene oxide coated mesoporous organic silicon oxide obtained in the step (1) with glucose oxidase, and performing condensation reaction to obtain the sequential nano catalyst.
2. The preparation method of the graphene oxide coated mesoporous organic silica according to claim 1, wherein the preparation method of the graphene oxide coated mesoporous organic silica in step (1) comprises the following steps:
1) Mixing a surfactant, a second solvent and a catalyst to obtain a mixed solution;
2) Mixing the mixed solution obtained in the step 1), a silicon source and 3-aminopropyl trimethoxy silane, and performing hydrolytic polycondensation reaction to obtain mesoporous organic silicon oxide;
3) And 2) mixing the mesoporous organic silicon oxide obtained in the step 2), water and graphene oxide solution, and performing electrostatic self-assembly to obtain the graphene oxide coated mesoporous organic silicon oxide.
3. The method according to claim 2, wherein the mass ratio of mesoporous organic silica to graphene oxide in step 3) is 2: (1-2).
4. The method of claim 1, wherein the activator in step (1) is EDC solution or NHS solution.
5. The process of claim 4, wherein the EDC solution and NHS solution are independently at a concentration of 1 to 2mg/mL.
6. The preparation method of claim 4, wherein in the step (1), the mass ratio of the graphene oxide coated mesoporous organic silica, EDC and NHS is 1: (1-1.2): (1-1.4).
7. The method according to claim 1, wherein the temperature of the carboxyl group activation treatment in the step (1) is room temperature, and the time of the carboxyl group activation treatment is 2 to 3 hours.
8. The preparation method according to claim 1, wherein the mass ratio of the graphene oxide coated mesoporous organic silica in the step (1) to the glucose oxidase in the step (2) is 1: (2-3).
9. The method according to claim 1, wherein the temperature of the condensation reaction in the step (2) is room temperature, and the time of the condensation reaction is 20 to 24 hours.
10. The sequential nanocatalyst prepared by the method of any of claims 1-9.
CN202410277726.3A 2024-03-12 2024-03-12 Sequential nano catalyst and preparation method thereof Pending CN118165978A (en)

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