CN113172232A

CN113172232A - Preparation method and application of Cu-In-Zn-Se composite nano-microspheres

Info

Publication number: CN113172232A
Application number: CN202110386883.4A
Authority: CN
Inventors: 盛扬; 薛亚波; 陈斌; 许红涛
Original assignee: JIANGSU CHENGUANG PAINT CO Ltd
Current assignee: JIANGSU CHENGUANG PAINT CO Ltd
Priority date: 2021-04-12
Filing date: 2021-04-12
Publication date: 2021-07-27
Anticipated expiration: 2041-04-12
Also published as: CN113172232B

Abstract

The invention belongs to the technical field of nano antibacterial materials, and particularly relates to a preparation method and application of a Cu-In-Zn-Se composite nano microsphere, wherein the preparation method comprises the following steps: (1) preparing a mixed solution of cuprous iodide, indium acetate, zinc stearate, 1-octadecene, oleylamine and dodecyl mercaptan; (2) preparing a precursor solution of selenium, and mixing and heating selenium powder, tri-n-octyl phosphine, 1-octadecene and oleylamine for dissolving; (3) heating the mixed solution in the step (1) to be fully dissolved, then adding a selenium precursor solution into the mixed solution, heating the mixed solution to 200 ℃ and 280 ℃, and preserving the heat for 10-40min to obtain a reaction solution; (4) centrifuging the reaction solution, cleaning and dispersing the solid phase In chloroform to obtain a chloroform solution of Cu-In-Zn-Se nanoparticles; (5) uniformly mixing a chloroform solution of Cu-In-Zn-Se nano particles with a chloroform solution of poly (maleic anhydride-alt-1-octadecene) and then drying; (6) and (5) adding the product obtained in the step (5) into a sodium hydroxide solution, performing ultrasonic dispersion, and removing redundant ions by a dialysis method until the pH value is neutral.

Description

Preparation method and application of Cu-In-Zn-Se composite nano-microspheres

Technical Field

The invention belongs to the technical field of nano antibacterial materials, and particularly relates to a preparation method and application of a Cu-In-Zn-Se composite nano microsphere.

Background

The microorganisms are everywhere, and especially in various public places such as hospitals, stations, hotels and the like, the personnel are complex and frequently move, pathogenic bacteria are easily spread, and the threat to public health is brought. Since ultraviolet rays are heavily scattered after entering the atmosphere, daily sunlight is insufficient to sufficiently kill microorganisms, and therefore, an antibacterial measure must be taken to inhibit their mass propagation. It is well known that the widespread use of pharmaceutical antibacterial agents, such as various antibiotics, has led to the emergence of drug-resistant bacteria and superbacteria. The non-drug antibacterial agent has obvious advantages in the aspects of antibacterial drug resistance and broad-spectrum antibacterial property because the non-drug antibacterial agent is not an action mechanism of biochemical reaction. However, the traditional non-drug antibacterial agents also have a plurality of problems, such as potential heavy metal biotoxicity of silver ion antibacterial agents to human bodies.

Photocatalytic antibiosis is widely concerned as a new non-drug antibiosis technology, but the prior photodynamic antibacterial agent materials have some defects, which seriously restrict the further popularization and application of the materials. For example, TiO₂And ZnO nano powder has wider band gap, can only absorb purple light with the wavelength less than 400nm in sunlight, and has lower utilization rate of solar energy; the organic photosensitive micromolecules have poor light stability and cannot continuously generate the bacteriostatic effect.

To fully utilize the solar spectral width, the semiconductor band gap width is reduced, but the excited state energy (Eg) is still higher than that of oxygen>0.97 eV). The Cu-In-Zn-Se nano particle is a direct band gap semiconductor nano material and has the advantages of narrow band gap (theoretical 1.04eV), low toxicity, high light absorption coefficient and the like. Especially, the nano-material prepared by the method has excellent optical performance. Under the action of illumination, the Cu-In-Zn-Se nano particles have the function of protecting the environmentThe oxygen and water molecules in the solution are converted into active oxygen species (e.g., singlet oxygen)¹O₂Hydroxyl radical, etc.). These reactive oxygen species can kill bacteria by oxidation. And because the inorganic semiconductor material has incomparable structure and light stability of organic molecules, the inorganic semiconductor material can continuously act and keep long-acting antibacterial ability. However, no published report of the application of the Cu-In-Zn-Se quaternary alloy nanoparticles In the field of photodynamic antibacterial is found at present.

The copper, zinc and selenium elements are rich in the earth crust. With the continuous improvement of the recovery and purification technology, the supply industry of indium is relatively stable. Therefore, the development of the photodynamic antibacterial agent based on the Cu-In-Zn-Se nano particles has important application value for solving the drug resistance of microorganisms and realizing long-acting antibiosis.

Disclosure of Invention

In order to solve the problems, the invention discloses a preparation method and application of a Cu-In-Zn-Se composite nano microsphere, wherein the preparation method comprises the steps of adding a Se precursor solution into a mixed solution containing Cu, In and Zn, heating for reaction, separating to obtain Cu-In-Zn-Se alloy nano particles, uniformly mixing a chloroform solution of the Cu-In-Zn-Se alloy nano particles with a chloroform solution of poly (maleic anhydride-alt-1-octadecene), drying, adding a sodium hydroxide solution into a dried product for dispersion, dialyzing to obtain an aqueous solution of the Cu-In-Zn-Se composite nano microsphere, the preparation method is simple, the Cu-In-Zn-Se composite nano microsphere can be stably dispersed In water, does not generate agglomeration, and can play a lasting role under visible light catalysis, Has antibacterial effect.

In order to achieve the purpose, the invention adopts the following technical scheme:

a preparation method of Cu-In-Zn-Se composite nano microspheres comprises the following steps:

(1) preparing a mixed solution of cuprous iodide, indium acetate, zinc stearate, 1-octadecene, oleylamine and dodecanethiol;

(2) preparing a precursor solution of selenium, mixing selenium powder, tri-n-octyl phosphine, 1-octadecene and oleylamine, and heating to dissolve;

(3) heating the mixed solution prepared in the step (1) in a protective atmosphere until the mixed solution is fully dissolved, then adding the selenium precursor solution prepared in the step (2), heating to 200-280 ℃, and preserving the temperature for 10-40min to obtain a reaction solution;

(4) cooling the reaction solution obtained In the step (3), centrifuging, cleaning the obtained solid phase, and dispersing In chloroform to obtain a chloroform solution of Cu-In-Zn-Se alloy nanoparticles;

(5) uniformly mixing the chloroform solution of the Cu-In-Zn-Se alloy nanoparticles obtained In the step (4) with the chloroform solution of poly (maleic anhydride-alt-1-octadecene), and drying;

(6) and (3) adding the product obtained In the step (5) into a sodium hydroxide solution, performing ultrasonic dispersion, and then removing redundant ions by a dialysis method until the pH value is neutral to obtain an aqueous solution of the Cu-In-Zn-Se composite nano microsphere.

Specifically, the molar ratio of cuprous iodide to indium acetate in step (1) is 1: 3.

Specifically, the molar ratio of cuprous iodide to zinc stearate in step (1) is 1: 1.

Specifically, the selenium precursor solution in the step (3) is injected at one time; the solvent used for cleaning the solid phase in the step (4) is ethanol or isopropanol.

Specifically, In the step (5), the mass ratio of the Cu-In-Zn-Se alloy nanoparticles to the poly (maleic anhydride-alt-1-octadecene) is 1:5-1: 10.

Specifically, the drying manner in the step (5) is normal temperature drying or heating drying.

Specifically, the concentration of the sodium hydroxide solution in the step (6) is 0.05-0.1 mol/L.

An application of a Cu-In-Zn-Se composite nano microsphere, wherein the Cu-In-Zn-Se composite nano microsphere is used as a photodynamic antibacterial agent.

Specifically, the application method of the Cu-In-Zn-Se composite nano microsphere comprises the following steps: the Cu-In-Zn-Se composite nano microspheres are dispersed In water to prepare a bacteriostatic agent solution with the concentration of 0.01-5.0 mg/mL.

Specifically, the Cu-In-Zn-Se composite nano microsphere has photocatalytic antibacterial property under visible light, and has long-acting photostability and antibacterial property.

The invention has the following beneficial effects:

(1) according to the preparation method of the Cu-In-Zn-Se composite nano microsphere, a precursor solution of Se is added into a mixed solution containing Cu, In and Zn, a Cu-In-Zn-Se alloy nano particle is obtained through separation after heating reaction, then a chloroform solution of the Cu-In-Zn-Se alloy nano particle and a chloroform solution of poly (maleic anhydride-alt-1-octadecene) are uniformly mixed and dried, and a dried product is added with a sodium hydroxide solution for dispersion and dialysis to obtain an aqueous solution of the Cu-In-Zn-Se composite nano microsphere; the Cu-In-Zn-Se composite nano-microspheres can be stably dispersed In water without agglomeration and can play a lasting and effective antibacterial role under the catalysis of visible light;

(2) the preparation method of the Cu-In-Zn-Se composite nano-microspheres optimizes the dosage ratio of each component, improves the yield of active oxygen substances and enhances the photodynamic antibacterial effect; the reaction temperature is higher, the crystallinity of the prepared nano particles can be improved, and the product quality is also improved;

(3) according to the invention, Cu-In-Zn-Se nano particles are compounded with poly (maleic anhydride-alt-1-octadecene) In chloroform, so that inorganic Cu-In-Zn-Se nano particles can be transferred from an oil phase to a water phase, and the obtained composite nano microspheres can keep stable dispersion In water and do not agglomerate;

(4) the Cu-In-Zn-Se composite nano-microsphere has good antibacterial performance, and compared with a photodynamic antibacterial agent prepared from organic photosensitive molecules, the Cu-In-Zn-Se composite nano-microsphere has good photostability, and the optical performance is only slightly reduced under long-time exposure of visible light. Therefore, the Cu-In-Zn-Se composite nano-microsphere has wide application prospect In the field of photodynamic antibiosis;

(5) the Cu-In-Zn-Se composite nano microsphere aqueous solution prepared by the invention has very good antibacterial effect on escherichia coli, and can reach an antibacterial rate of 90% at 500 mug/mL.

Drawings

The invention is further illustrated with reference to the following figures and examples.

FIG. 1 is a fluorescence spectrum of Cu-In-Zn-Se nanoparticles prepared In example 1 of Cu-In-Zn-Se nanoparticles and comparative examples 1 to 4 of Cu-In-Zn-Se nanoparticles;

FIG. 2 is CuIn prepared from Cu-In-Zn-Se nanoparticles example 1₃ZnSe₆A plot of the fluorescence intensity of the nanoparticles as a function of time under visible light exposure;

FIG. 3 is a graph of the change of absorbance of singlet oxygen indicator 9, 10-anthryl-bis (methylene) dipropionic acid (ADMA) with time under the action of light caused by Cu-In-Zn-Se composite nano microspheres prepared In composite nano microsphere example 1 and composite nano microsphere comparative examples 1-4;

FIG. 4 shows the antibacterial efficiency of the Cu-In-Zn-Se composite nanospheres prepared In the composite nanosphere example 1 and the composite nanosphere comparative examples 1-4 on Escherichia coli after being diluted with different concentrations of aqueous solutions.

Detailed Description

To further clarify the objects, technical solutions and advantages of the present invention, the present invention will now be further described with reference to specific examples, which are intended to illustrate the invention without further limiting it.

Cu-In-Zn-Se nanoparticles example 1

Preparation of CuIn₃ZnSe₆The method comprises the following specific steps of:

(1) weighing 1mmol of cuprous iodide, 3mmol of indium acetate, 1mmol of zinc stearate, 12mL of 1-octadecene, 1mL of dodecanethiol and 4mL of oleylamine, and preparing a mixed solution;

(2) weighing 8mmol of selenium powder, 1mL of 1-octadecene, 1mL of tri-n-octylphosphine and 0.3mL of oleylamine, and heating to 120 ℃ for dissolving to obtain a precursor solution of selenium;

(3) transferring the mixed solution prepared in the step (1) to a 50mL three-neck flask, heating to 120 ℃ under the protection of nitrogen to fully dissolve the mixed solution, then adding the precursor solution of selenium prepared in the step (2) into the mixed solution at one time, continuously heating the solution to 250 ℃, keeping the temperature for 20min, and cooling to obtain a reaction solution;

(4) and (4) cooling the reaction solution obtained In the step (3), centrifuging, washing the solid phase with ethanol for three times to obtain a brown solid, and dispersing the brown solid In chloroform according to the concentration of 10mg/mL to obtain a chloroform solution of the Cu-In-Zn-Se alloy nanoparticles.

Cu-In-Zn-Se nanoparticle comparative example 1

Cu-In-Zn-Se nanoparticle comparative example 1 is different from Cu-In-Zn-Se nanoparticle example 1 In that: comparative example 1 of Cu-In-Zn-Se nanoparticles cuprous iodide was 2mmol, indium acetate was 2mmol In step (1), and the remainder was the same as In example 1 of Cu-In-Zn-Se nanoparticles to give 10mg/mL of CuInZn_0.5Se_2.5Chloroform solution of nanoparticles.

Cu-In-Zn-Se nanoparticle comparative example 2

Cu-In-Zn-Se nanoparticle comparative example 2 is different from Cu-In-Zn-Se nanoparticle example 1 In that: Cu-In-Zn-Se nanoparticles comparative example 2 copper iodide 3.0mmol and indium acetate 1.0mmol In step (1), and the remainder as Cu-In-Zn-Se nanoparticles example 1, to give 10mg/mL Cu₃InZnSe₄Chloroform solution of nanoparticles.

Comparative Cu-In-Zn-Se nanoparticle example 3

Cu-In-Zn-Se nanoparticle comparative example 3 is different from Cu-In-Zn-Se nanoparticle example 1 In that: Cu-In-Zn-Se nanoparticles comparative example 3 Zinc stearate In step (1) was 0.5mmol, the remainder as Cu-In-Zn-Se nanoparticles example 1, 10mg/mL CuIn₃Zn_0.5Se_5.5Chloroform solution of nanoparticles.

Comparative Cu-In-Zn-Se nanoparticle example 4

Cu-In-Zn-Se nanoparticle comparative example 4 is different from Cu-In-Zn-Se nanoparticle example 1 In that: Cu-In-Zn-Se nanoparticles comparative example 4 Zinc stearate In step (1) was 2.0mmol, the remainder as Cu-In-Zn-Se nanoparticles example 1, 10mg/mL CuIn₃Zn₂Se₇Chloroform solution of nanoparticles.

As can be seen from the fluorescence spectrum of FIG. 1, the CuIn of Cu-In-Zn-Se nanoparticle example 1₃ZnSe₆The nanoparticles have stronger fluorescence intensity, which indicates less defects and less heat loss, and the quantum yield measured by a comparative method is about 37%. The Cu-In-Zn-Se nanoparticle comparative example 1 and the Cu-In-Zn-Se nanoparticle comparative example 2 gradually decreased the fluorescence intensity as the Cu ratio increased, indicating more heat loss. Measured quantum yield of-2% and-15%. The decrease in quantum yield indicates that it is useful for catalyzing O₂To be converted into¹O₂The efficiency of (a) is lowered, and the yield of the active oxygen species is reduced, making it difficult to achieve the antibacterial effect of Cu-In-Zn-Se nanoparticle example 1. The Cu-In-Zn-Se nanoparticle comparative example 3 and the Cu-In-Zn-Se nanoparticle comparative example 4 have decreased fluorescence intensity and decreased quantum yield to-20% and-5% when the amount of Zn doping was decreased or increased, compared to the Cu-In-Zn-Se nanoparticle example 1, and thus it was also difficult to achieve the antibacterial effect of the Cu-In-Zn-Se nanoparticle example 1.

As can be seen from FIG. 2, the CuIn prepared In Cu-In-Zn-Se nanoparticle example 1₃ZnSe₆The nanoparticles have good photostability and photobleaching resistance. After continuous irradiation with visible light for about 2000 hours, the fluorescence intensity is reduced by less than 10%, which proves that the original structure is remained and is not degraded or decomposed.

Composite Nanoparballoball example 1

The preparation method of the Cu-In-Zn-Se composite nano microsphere comprises the following specific steps:

(1) dissolving 100mg of poly (maleic anhydride-alt-1-octadecene) in 10mL of chloroform at a concentration of 10 mg/mL;

(2) taking 1 part of chloroform solution of Cu-In-Zn-Se nanoparticles prepared In Cu-In-Zn-Se nanoparticle example 1 and 9 parts of chloroform solution of poly (maleic anhydride-alt-1-octadecene), uniformly mixing, and removing a solvent to obtain dry solid powder;

(3) adding 90mL of 0.08M NaOH solution into the solid powder, performing ultrasonic treatment for about 1h to fully disperse the solid powder, dialyzing for 12h by using a 8000Da dialysis membrane, replacing the dialyzed solution every 3h, and diluting with sterile water to obtain a Cu-In-Zn-Se composite nano microsphere aqueous solution with the concentration of 10mg/mL after dialysis.

Comparative example 1 of composite nanospheres

Substantially the same as In composite nanosphere example 1 except that a chloroform solution of Cu-In-Zn-Se nanoparticles prepared In Cu-In-Zn-Se nanoparticle comparative example 1 was used In step (2) of composite nanosphere comparative example 1.

Comparative example 2 of composite nanospheres

Substantially the same as In composite nanosphere example 1 except that a chloroform solution of Cu-In-Zn-Se nanoparticles prepared In Cu-In-Zn-Se nanoparticle comparative example 2 was used In step (2) of composite nanosphere comparative example 2.

Comparative example 3 of composite nanospheres

Substantially the same as In example 1 except that a chloroform solution of Cu-In-Zn-Se nanoparticles prepared In comparative example 3 was used In step (2) of comparative example 3.

Comparative example 4 of composite nanospheres

Substantially the same as In composite nanosphere example 1 except that a chloroform solution of Cu-In-Zn-Se nanoparticles prepared In Cu-In-Zn-Se nanoparticle comparative example 4 was used In step (2) of composite nanosphere comparative example 4.

FIG. 3 is¹O₂The absorbance of the indicator ADMA changes along with time under the photocatalysis of the Cu-In-Zn-Se composite nano microspheres. Due to CuIn₃ZnSe₆Has higher quantum yield, so more excited-state electrons can react with dissolved oxygen In the environment to generate In comparison with other Cu-In-Zn-Se nano particles¹O₂And therefore has the highest¹O₂Yield, the fastest decrease in absorbance of ADMA. Since the Cu-In-Zn-Se nanoparticles have good photostability, the Cu-In-Zn-Se nanoparticles can be continuously generated¹O₂Degrading ADMA. ADMA can still be rapidly degraded after 4 hours of continuous light irradiation.

Composite Nanoparballoball example 2

(2) taking 1 part of chloroform solution of Cu-In-Zn-Se nanoparticles prepared In Cu-In-Zn-Se nanoparticle example 1 and 10 parts of chloroform solution of poly (maleic anhydride-alt-1-octadecene), uniformly mixing, and removing a solvent to obtain dry solid powder;

(3) adding 90mL of 0.1M NaOH solution into the solid powder, performing ultrasonic treatment for about 1h to fully disperse the solid powder, dialyzing for 12h by using a 8000Da dialysis membrane, replacing the dialyzed solution every 3h, and diluting with sterile water to obtain a Cu-In-Zn-Se composite nano microsphere aqueous solution with the concentration of 10mg/mL after dialysis.

Composite Nanoparballoball example 3

(2) taking 1 part of chloroform solution of Cu-In-Zn-Se nanoparticles prepared In Cu-In-Zn-Se nanoparticle example 1 and 5 parts of chloroform solution of poly (maleic anhydride-alt-1-octadecene), uniformly mixing, and removing a solvent to obtain dry solid powder;

(3) adding 90mL of 0.05M NaOH solution into the solid powder, performing ultrasonic treatment for about 1h to fully disperse the solid powder, dialyzing for 12h by using a 8000Da dialysis membrane, replacing the dialyzed solution every 3h, and diluting with sterile water to obtain a Cu-In-Zn-Se composite nano microsphere aqueous solution with the concentration of 10mg/mL after dialysis.

Application example

Inhibition of the Cu-In-Zn-Se composite nano-microspheres on escherichia coli:

(1) escherichia coli (ATCC,25922) was added to the sterilized LB liquid medium, incubated overnight on a shaker (37 ℃ C., 250rpm), and then diluted to 1X 10 with the sterilized LB liquid medium⁵CFU/mL, then added to 48-well plates (200. mu.L/well);

(2) diluting 10mg/mL of Cu-In-Zn-Se composite nanoparticle aqueous solutions prepared In composite nanoparticle example 1 and composite nanoparticle comparative examples 1-4 to different concentrations (10. mu.g/mL, 20. mu.g/mL, 50. mu.g/mL, 100. mu.g/mL, 200. mu.g/mL, 500. mu.g/mL, 1mg/mL, 2mg/mL, 5mg/mL) by using sterile water, adding 48-well plates (200. mu.L/well) of step (1), placing In a shaking table (37 ℃, 250rpm) for illumination culture for 8h, and taking out; mu.L of PBS (pH 7.4, 10mM) was added to 200. mu.L (concentration 1X 10) of E.coli⁵CFU/mL), culturing in dark environmentAfter 8h, the sample was taken out as a control.

(3) Respectively diluting 10 the Cu-In-Zn-Se composite nano microsphere aqueous solution cultured In the step (2) by using sterile water⁵After doubling, 100. mu.L of the suspension was uniformly applied to LB solid medium, and after 1 day of culture in an incubator at 37 ℃, the number of viable bacteria was observed and the antibacterial efficiency was calculated.

FIG. 4 is a comparison of antibacterial effects of Cu-In-Zn-Se composite nano microsphere aqueous solutions with different concentrations and different compositions on Escherichia coli. Due to CuIn₃ZnSe₆Has the highest¹O₂The yield and therefore the antibacterial effect are also best, and the antibacterial rate can reach 90 percent at 500 mu g/mL.

In light of the foregoing description of the preferred embodiment of the present invention, many modifications and variations will be apparent to those skilled in the art without departing from the spirit and scope of the invention. The technical scope of the present invention is not limited to the content of the specification, and must be determined according to the scope of the claims.

Claims

1. A preparation method of Cu-In-Zn-Se composite nano microspheres is characterized by comprising the following steps: the method comprises the following steps:

2. The method for preparing Cu-In-Zn-Se composite nanospheres according to claim 1, wherein: the molar ratio of cuprous iodide to indium acetate in the step (1) is 1: 3.

3. The method for preparing Cu-In-Zn-Se composite nanospheres according to claim 1, wherein: the molar ratio of cuprous iodide to zinc stearate in the step (1) is 1: 1.

4. The method for preparing Cu-In-Zn-Se composite nanospheres according to claim 1, wherein: the selenium precursor solution in the step (3) is injected at one time; the solvent used for cleaning the solid phase in the step (4) is ethanol or isopropanol.

5. The method for preparing Cu-In-Zn-Se composite nanospheres according to claim 1, wherein: in the step (5), the mass ratio of the Cu-In-Zn-Se alloy nanoparticles to the poly (maleic anhydride-alt-1-octadecene) is 1:5-1: 10.

6. The method for preparing Cu-In-Zn-Se composite nanospheres according to claim 1, wherein: and (5) drying at normal temperature or heating.

7. The method for preparing Cu-In-Zn-Se composite nanospheres according to claim 1, wherein: the concentration of the sodium hydroxide solution in the step (6) is 0.05-0.1 mol/L.

8. Use of Cu-In-Zn-Se composite nanospheres according to any of claims 1 to 7, characterized In that: the Cu-In-Zn-Se composite nano-microsphere is used as a photodynamic antibacterial agent.

9. The use of Cu-In-Zn-Se composite nanospheres according to claim 8, wherein: the application method comprises the following steps: the Cu-In-Zn-Se composite nano microspheres are dispersed In water to prepare a bacteriostatic agent solution with the concentration of 0.01-5.0 mg/mL.

10. The use of Cu-In-Zn-Se composite nanospheres according to claim 8, wherein: the Cu-In-Zn-Se composite nano microsphere has photocatalytic antibacterial property under visible light and long-acting photostability and antibacterial property.