Use of a cuprous phosphide

Abstract

The application relates to an application of cuprous phosphide and belongs to the technical field of cuprous phosphide application. The application solves the problems that the antibacterial performance of an antibacterial material is low in the prior art, the antibacterial material depends on external conditions such as light in the use process, the preparation method of the antibacterial material is complex, the condition is harsh, and the cost is high. The application provides the application of cuprous phosphide as the antibacterial material, the application is simple, super-efficient, has a broad-spectrum bactericidal effect, and does not depend on external auxiliary conditions in the use process, and does not need a complex and harsh preparation method.

Description

Technical Field

[0001] This invention belongs to the field of cuprous phosphide application technology, and specifically relates to an application of cuprous phosphide. Background Technology

[0002] Bacterial diseases are infectious or contagious illnesses caused by bacteria. Clinically, the most effective treatment for bacterial diseases is antibiotics. However, long-term and continuous use of antibiotics has led to the emergence of drug-resistant bacteria.

[0003] To address this technical challenge, researchers have developed various nanomaterials with unique antibacterial mechanisms as novel antibacterial agents. Among existing technologies, the antibacterial application of silver nanoparticles is a successful example. In recent years, other nanomaterials with unique physicochemical properties have also been developed for antibacterial applications. Mechanisms of action include inducing bacterial cell membrane or cell wall disruption through physical contact between the material and bacteria, thereby killing the bacteria, or using auxiliary conditions such as light / heat to mediate antibacterial action. Furthermore, nanomaterials with enzyme-like activity (also known as enzyme mimics or nanozymes), particularly peroxidase-like and oxidase-like materials, have been developed as novel antibacterial materials due to their ability to catalyze the conversion of oxygen or hydrogen peroxide into reactive oxygen species, exhibiting excellent antibacterial properties.

[0004] These research findings demonstrate the enormous potential of nanomaterials as antibiotic alternatives, but existing antibacterial materials still have some shortcomings. (1) Excessive release of silver ions poses potential hazards to the environment and human health, and the use of precious metals increases costs. (2) Physical sterilization is less efficient, requiring higher concentrations or longer durations, which limits its practical application. (3) Antibacterial applications mediated by auxiliary conditions such as light / heat are condition-dependent in practical use. (4) Most oxidase-like and peroxidase-like mimics require acidic conditions (pH 3-4) for optimal reaction, limiting their application in antibacterial applications. Therefore, developing new antibacterial materials that can overcome the above limitations as antibiotic alternatives remains both necessary and challenging.

[0005] Cuprous phosphide, Cu3P, is produced by the reaction of copper and phosphorus. It is grayish-yellow in color, brittle, does not react with water, is insoluble in dilute inorganic acids, but readily soluble in strong oxidizing acids, undergoing redox reactions. In current technologies, the applications of cuprous phosphide are mainly concentrated in photocatalysis, electrocatalysis, and lithium-ion / sodium-ion batteries. One existing technology reports a Cu3P nanowire-modified copper mesh electrode for the inactivation of pathogens in water. The essence of this technology is electric field sterilization, with Cu3P serving as a modifying material to enhance electrode function. The tip properties of Cu3P influence the electric field effect, and the mechanism of enhanced electric field sterilization is explained as electroporation (or electrical breakdown) (J. Mater. Chem. A, 2018, 6, 18813). The application of cuprous phosphide itself as an antibacterial material has not yet been discovered, and the related antibacterial mechanism has not been reported.

[0006] This invention provides an application of cuprous phosphide itself as an antibacterial material, which does not rely on electric field sterilization and requires no external auxiliary conditions. Furthermore, its antibacterial mechanism is explained in principle as follows: the material's enzyme-like properties generate reactive oxygen species, thereby endowing it with bactericidal properties. Simultaneously, the material mediates the depletion of glutathione within bacterial cells and possesses hydrolytic enzyme-like activity, increasing bacterial cell membrane permeability, thus enhancing the antibacterial effect. The bactericidal characteristics of this material overcome the limitations of aforementioned nano-antibacterial materials: it does not release metal ions, requires extremely low concentrations, requires no auxiliary conditions, and is effective across a wide pH range from acidic to neutral. Summary of the Invention

[0007] This invention addresses the problems of low antibacterial performance of existing antibacterial materials, reliance on external auxiliary conditions during use, and complex, demanding, and costly preparation methods. It provides an application of cuprous phosphide.

[0008] The technical solution adopted by the present invention to solve the above-mentioned technical problems is as follows.

[0009] This invention provides an application of cuprous phosphide as an antibacterial material.

[0010] Preferably, cuprous phosphide is applied in either a liquid phase or a solid phase.

[0011] Preferably, the liquid phase application involves adding cuprous phosphide to a liquid containing bacteria; the solid phase application involves placing cuprous phosphide on the surface of an object containing bacteria.

[0012] Preferably, in the liquid phase application, the minimum final concentration of cuprous phosphide to achieve an inhibition rate of over 99.9% against Gram-positive bacteria is 0.5 μg / mL. -1 and 1.5 μg mL -1 .

[0013] Preferably, in the liquid phase application, the minimum final concentration at which cuprous phosphide achieves an antibacterial rate of over 99.9% against natural water bodies is 0.8 μg / mL. -1 .

[0014] Preferably, the solid-phase application is to prepare cuprous phosphide into a washing solution and apply it to the surface of an object containing bacteria, or to prepare cuprous phosphide into a dressing and apply it to the surface of an object containing bacteria.

[0015] Preferably, the concentration of cuprous phosphide in the washing solution is 0.5 μg / mL. -1 Up to 40 μg mL -1 The amount of cuprous phosphide used in the dressing is 0.5 μg cm. -2 Up to 40 μg cm -2 .

[0016] Preferably, the antibacterial time is greater than 0 min, and more preferably, the antibacterial time with an inhibition rate of 99.9% or higher is greater than 20 min.

[0017] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0018] This invention provides the application of cuprous phosphide as an antibacterial material. This application is simple, highly efficient, and has a broad-spectrum bactericidal effect. It does not rely on auxiliary conditions such as light / heat during use and does not require complex or harsh preparation methods. Attached Figure Description

[0019] To more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these embodiments without creative effort.

[0020] Figure 1 These are TEM images of Cu3P used in Examples 1-19 of this invention;

[0021] Figure 2 The XRD patterns of Cu3P used in Examples 1-19 of this invention are shown.

[0022] Figure 3 The results of the antibacterial activity test of Cu3P in Examples 1-2 of this invention;

[0023] Figure 4 The results of the antibacterial activity test of Cu3P in Examples 3-12 of this invention;

[0024] Figure 5 The antibacterial test results of Cu3P in Example 13 of this invention;

[0025] Figure 6 The antibacterial test results of Cu3P in Example 14 of this invention;

[0026] Figure 7 The antibacterial test results of Cu3P in Example 15 of this invention;

[0027] Figure 8 The antibacterial test results of Cu3P in Example 16 of this invention;

[0028] Figure 9 The antibacterial test results of Cu3P in Example 17 of this invention;

[0029] Figure 10 The results of Cu3P cytotoxicity detection in Examples 18-19 of this invention are shown. Detailed Implementation

[0030] To further understand the present invention, preferred embodiments of the present invention are described below in conjunction with specific implementation methods. However, it should be understood that these descriptions are only for further illustrating the features and advantages of the present invention and not for limiting the claims of the present invention.

[0031] The cuprous phosphide of the present invention can be used as an antibacterial material. There are no special limitations on the specific application form, and it can be applied to various application forms of powdered antibacterial materials in the prior art, that is, the cuprous phosphide can be used in liquid phase or solid phase.

[0032] For liquid phase applications, cuprous phosphide is preferably added to the liquid containing bacteria. In liquid phase applications, the minimum final concentration of cuprous phosphide to achieve an inhibition rate of over 99.9% against Gram-positive bacteria is 0.5 μg / mL. -1 The minimum final concentration at which cuprous phosphide achieves an inhibition rate of over 99.9% against Gram-negative bacteria is 1.5 μg / mL. -1 The minimum final concentration at which cuprous phosphide achieves an antibacterial rate of over 99.9% in natural water bodies is 0.8 μg / mL. -1 There are no specific restrictions on the concentration of the bacterial strain in the liquid containing bacteria; the concentration can be 3 × 10⁻⁶. 6 CFU mL -1 the following.

[0033] Solid-phase application is preferably performed by placing cuprous phosphide on the surface of an object containing bacteria. Specifically, solid-phase application involves preparing a washing solution of cuprous phosphide and applying it to the surface of the object containing bacteria, or preparing a dressing of cuprous phosphide and applying it to the surface of the object containing bacteria. The concentration of cuprous phosphide in the washing solution is 0.5 μg / mL. -1 Up to 40 μg mL -1 The solute is distilled water. The amount of cuprous phosphide used in the dressing is 0.5 μg cm⁻¹.-2 Up to 40 μg cm -2 .

[0034] In this invention, the antibacterial time is preferably 5 minutes or more; more preferably 20 minutes or more; and most preferably 20 minutes.

[0035] The terminology used in this invention generally has the meanings commonly understood by those skilled in the art, unless otherwise stated. In this invention, significant antibacterial effect is defined as an inhibition rate of 99.9% or higher. Natural water bodies refer to the natural complexes of the Earth's surface covered by water, including oceans, rivers, lakes, swamps, and glaciers. Water bodies not only refer to water itself but also include various substances contained within them, such as dissolved substances, suspended substances, aquatic organisms, and substrate. Objects refer to all tangible matter that objectively exists in nature, including the human body.

[0036] To enable those skilled in the art to better understand the technical solutions of the present invention, the present invention will be further described in detail below with reference to embodiments.

[0037] In the following examples, various processes and methods not described in detail are conventional methods known in the art. Unless otherwise specified, the materials, reagents, apparatus, instruments, and equipment used in the following examples are commercially available. Cuprous phosphide was obtained by phosphating copper oxide with sodium hypophosphite at a reaction temperature of 300°C for 2 hours, a heating rate of 5°C / min, and a mass ratio of sodium hypophosphite to copper oxide of 15:1. Copper oxide was prepared according to the methods described in the literature (Konar, S.; Kalita, H.; Puvvada, N.; Tantubay, S.; Mahto, MK; Biswas, S.; Pathak, A., Shape-dependent catalytic activity of CuO nanostructures. Journal of Catalysis 2016, 336, 11-22.).

[0038] The present invention will be further illustrated below with reference to the embodiments.

[0039] Example 1

[0040] Step 1: Transfer a single Gram-negative Escherichia coli (E. coli) colony to 20 mL of Luria-Bertani (LB) medium and incubate at 37°C in a shaker for 12 h. Then, dilute to 3 × 10⁻⁶ with sterile PBS buffer at pH 7.4. 6 CFUmL -1 A bacterial solution was obtained. A final concentration of 1.5 μg / mL was added to the bacterial solution. -1Cu3P (morphology as) Figure 1 As shown in (a), the spherical nanoparticles have a particle size of approximately 35 nm, and the XRD pattern is as follows. Figure 2 (As shown in curve (a)). After treatment for 0 min, 5 min, 10 min, and 20 min respectively, the cells were diluted with sterile PBS buffer at pH 7.4 to a colony concentration of 10. 3 CFU mL -1 The bacterial dilution was obtained. Finally, 100 μL of the bacterial dilution was evenly spread onto LB agar plates and incubated at 37°C for 24 h. The number of colonies was then counted using the plate count method. The results are as follows: Figure 3 As shown.

[0041] from Figure 3 It can be seen that the concentration is 1.5 μg / mL -1 Cu3P can achieve a bactericidal efficiency of 99.9% against Gram-negative bacteria Escherichia coli (E. coli) after 20 min.

[0042] Example 2

[0043] A single Gram-positive methicillin-resistant Staphylococcus aureus (MRSA) was transferred to 20 mL of Luria-Bertani (LB) medium and incubated for 12 h in a shaker at 37 °C. The culture was then diluted to 3 × 10⁻⁶ with sterile PBS buffer at pH 7.4. 6 CFU mL -1 A bacterial solution was obtained. Adding to the bacterial solution a final concentration of 0.5 μg / mL... -1 Cu3P (morphology as) Figure 1 As shown in (a), the spherical nanoparticles have a particle size of approximately 35 nm, and the XRD pattern is as follows. Figure 2 (As shown in curve (a)). After treatment for 0 min, 5 min, 10 min, and 20 min respectively, the cells were diluted with PBS to a colony concentration of 10. 3 CFU mL -1 The bacterial dilution was obtained. Finally, 100 μL of the bacterial dilution was evenly spread onto LB agar plates and incubated at 37°C for 24 h. The number of colonies was then counted using the plate count method. The results are as follows: Figure 3 As shown.

[0044] from Figure 3 It can be seen that the concentration is 0.5 μg / mL -1 Cu3P achieved a bactericidal efficiency of 99.9% against methicillin-resistant Staphylococcus aureus (MRSA) in 20 minutes.

[0045] Example 3

[0046] Step 1: Transfer a single Gram-negative Escherichia coli (E. coli) colony to 20 mL of Luria-Bertani (LB) medium and incubate at 37°C in a shaker for 12 h. Then, dilute to 3 × 10⁻⁶ with sterile PBS buffer at pH 7.4. 6 CFUmL -1 A bacterial solution was obtained. A final concentration of 1.5 μg / mL was added to the bacterial solution. -1 Cu3P (morphology as) Figure 1 As shown in (b), the particles have an irregular morphology and a size of approximately 50 nm. The XRD pattern is as follows. Figure 2 (As shown in curve (b)). After treatment for 0 min and 20 min respectively, the cells were diluted with sterile PBS buffer at pH 7.4 to a colony concentration of 10. 3 CFU mL -1 The bacterial dilution was obtained. Finally, 100 μL of the bacterial dilution was evenly spread onto LB agar plates and incubated at 37°C for 24 h. The number of colonies was then counted using the plate count method. The results are as follows: Figure 4 As shown.

[0047] from Figure 4 It can be seen that the concentration is 1.5 μg / mL -1 Cu3P can achieve a bactericidal efficiency of 99.9% against Gram-negative bacteria Escherichia coli (E. coli) after 20 min.

[0048] Example 4

[0049] A single Gram-positive methicillin-resistant Staphylococcus aureus (MRSA) was transferred to 20 mL of Luria-Bertani (LB) medium and incubated for 12 h in a shaker at 37 °C. The culture was then diluted to 3 × 10⁻⁶ with sterile PBS buffer at pH 7.4. 6 CFU mL -1 A bacterial solution was obtained. A final concentration of 0.5 μg / mL was added to the bacterial solution. -1 Cu3P (morphology as) Figure 1 As shown in (b), the particles have an irregular morphology and a size of approximately 50 nm. The XRD pattern is as follows. Figure 2 (As shown in curve (b)). After treatment for 0 min and 20 min respectively, the cells were diluted with PBS to a colony concentration of 10. 3 CFU mL -1 The bacterial dilution was obtained. Finally, 100 μL of the bacterial dilution was evenly spread onto LB agar plates and incubated at 37°C for 24 h. The number of colonies was then counted using the plate count method. The results are as follows: Figure 4 As shown.

[0050] from Figure 4 It can be seen that the concentration is 0.5 μg / mL -1 Cu3P achieved a bactericidal efficiency of 99.9% against methicillin-resistant Staphylococcus aureus (MRSA) in 20 minutes.

[0051] Example 5

[0052] Step 1: Transfer a single Gram-negative Escherichia coli (E. coli) colony to 20 mL of Luria-Bertani (LB) medium and incubate at 37°C in a shaker for 12 h. Then, dilute to 3 × 10⁻⁶ with sterile PBS buffer at pH 7.4. 6 CFUmL -1 A bacterial solution was obtained. A final concentration of 1.5 μg / mL was added to the bacterial solution. -1 Cu3P (morphology as) Figure 1 As shown in (c), the spherical nanoparticles have a particle size of approximately 100 nm, and the XRD pattern is as follows. Figure 2 (As shown in curve (c)). After treatment for 0 min and 20 min respectively, the cells were diluted with sterile PBS buffer at pH 7.4 to a colony concentration of 10. 3 CFU mL -1 The bacterial dilution was obtained. Finally, 100 μL of the bacterial dilution was evenly spread onto LB agar plates and incubated at 37°C for 24 h. The number of colonies was then counted using the plate count method. The results are as follows: Figure 4 As shown.

[0053] from Figure 4 It can be seen that the concentration is 1.5 μg / mL -1 Cu3P can achieve a bactericidal efficiency of 99.9% against Gram-negative bacteria Escherichia coli (E. coli) after 20 min.

[0054] Example 6

[0055] A single Gram-positive methicillin-resistant Staphylococcus aureus (MRSA) was transferred to 20 mL of Luria-Bertani (LB) medium and incubated for 12 h in a shaker at 37 °C. The culture was then diluted to 3 × 10⁻⁶ with sterile PBS buffer at pH 7.4. 6 CFU mL -1 A bacterial solution was obtained. A final concentration of 0.5 μg / mL was added to the bacterial solution. -1 Cu3P (morphology as) Figure 1 As shown in (c), the spherical nanoparticles have a particle size of approximately 100 nm, and the XRD pattern is as follows. Figure 2 (As shown in curve (c)). After treatment for 0 min and 20 min respectively, the cells were diluted with PBS to a colony concentration of 10.3 CFU mL -1 The bacterial dilution was obtained. Finally, 100 μL of the bacterial dilution was evenly spread onto LB agar plates and incubated at 37°C for 24 h. The number of colonies was then counted using the plate count method. The results are as follows: Figure 4 As shown.

[0056] from Figure 4 It can be seen that the concentration is 0.5 μg / mL -1 Cu3P achieved a bactericidal efficiency of 99.9% against methicillin-resistant Staphylococcus aureus (MRSA) in 20 minutes.

[0057] Example 7

[0058] Step 1: Transfer a single Gram-negative Escherichia coli (E. coli) colony to 20 mL of Luria-Bertani (LB) medium and incubate at 37°C in a shaker for 12 h. Then, dilute to 3 × 10⁻⁶ with sterile PBS buffer at pH 7.4. 6 CFUmL -1 A bacterial solution was obtained. A final concentration of 1.5 μg / mL was added to the bacterial solution. -1 Cu3P (morphology as) Figure 1 As shown in (d), the spherical nanoparticles have a particle size of approximately 400 nm, and the XRD pattern is as follows. Figure 2 (As shown in curve (d)). After treatment for 0 min and 20 min respectively, the cells were diluted with sterile PBS buffer at pH 7.4 to a colony concentration of 10. 3 CFU mL -1 The bacterial dilution was obtained. Finally, 100 μL of the bacterial dilution was evenly spread onto LB agar plates and incubated at 37°C for 24 h. The number of colonies was then counted using the plate count method. The results are as follows: Figure 4 As shown.

[0059] from Figure 4 It can be seen that the concentration is 1.5 μg / mL -1 Cu3P can achieve a bactericidal efficiency of 99.9% against Gram-negative bacteria Escherichia coli (E. coli) after 20 min.

[0060] Example 8

[0061] A single Gram-positive methicillin-resistant Staphylococcus aureus (MRSA) was transferred to 20 mL of Luria-Bertani (LB) medium and incubated for 12 h in a shaker at 37 °C. The culture was then diluted to 3 × 10⁻⁶ with sterile PBS buffer at pH 7.4. 6 CFU mL -1A bacterial solution was obtained. A final concentration of 0.5 μg / mL was added to the bacterial solution. -1 Cu3P (morphology as) Figure 1 As shown in (d), the spherical nanoparticles have a particle size of approximately 400 nm, and the XRD pattern is as follows. Figure 2 (As shown in curve (d)). After treatment for 0 min and 20 min respectively, the cells were diluted with PBS to a colony concentration of 10. 3 CFU mL -1 The bacterial dilution was obtained. Finally, 100 μL of the bacterial dilution was evenly spread onto LB agar plates and incubated at 37°C for 24 h. The number of colonies was then counted using the plate count method. The results are as follows: Figure 4 As shown.

[0062] from Figure 4 It can be seen that the concentration is 0.5 μg / mL -1 Cu3P achieved a bactericidal efficiency of 99.9% against methicillin-resistant Staphylococcus aureus (MRSA) in 20 minutes.

[0063] Example 9

[0064] Step 1: Transfer a single Gram-negative Escherichia coli (E. coli) colony to 20 mL of Luria-Bertani (LB) medium and incubate at 37°C in a shaker for 12 h. Then, dilute to 3 × 10⁻⁶ with sterile PBS buffer at pH 7.4. 6 CFUmL -1 A bacterial solution was obtained. A final concentration of 1.5 μg / mL was added to the bacterial solution. -1 Cu3P (morphology as) Figure 1 As shown in (e), the spherical nanoparticles have a particle size of approximately 30 nm, and the XRD pattern is as follows. Figure 2 (As shown in curve (e)). After treatment for 0 min and 20 min respectively, the cells were diluted with sterile PBS buffer at pH 7.4 to a colony concentration of 10. 3 CFU mL -1 The bacterial dilution was obtained. Finally, 100 μL of the bacterial dilution was evenly spread onto LB agar plates and incubated at 37°C for 24 h. The number of colonies was then counted using the plate count method. The results are as follows: Figure 4 As shown.

[0065] from Figure 4 It can be seen that the concentration is 1.5 μg / mL -1 Cu3P can achieve a bactericidal efficiency of 99.9% against Gram-negative bacteria Escherichia coli (E. coli) after 20 min.

[0066] Example 10

[0067] A single Gram-positive methicillin-resistant Staphylococcus aureus (MRSA) was transferred to 20 mL of Luria-Bertani (LB) medium and incubated for 12 h in a shaker at 37 °C. The culture was then diluted to 3 × 10⁻⁶ with sterile PBS buffer at pH 7.4. 6 CFU mL -1 A bacterial solution was obtained. A final concentration of 0.5 μg / mL was added to the bacterial solution. -1 Cu3P (morphology as) Figure 1 As shown in (e), the spherical nanoparticles have a particle size of approximately 30 nm, and the XRD pattern is as follows. Figure 2 (As shown in curve (e)). After treatment for 0 min and 20 min respectively, the cells were diluted with PBS to a colony concentration of 10. 3 CFU mL -1 The bacterial dilution was obtained. Finally, 100 μL of the bacterial dilution was evenly spread onto LB agar plates and incubated at 37°C for 24 h. The number of colonies was then counted using the plate count method. The results are as follows: Figure 4 As shown.

[0068] from Figure 4 It can be seen that the concentration is 0.5 μg / mL -1 Cu3P achieved a bactericidal efficiency of 99.9% against methicillin-resistant Staphylococcus aureus (MRSA) in 20 minutes.

[0069] Example 11

[0070] Step 1: Transfer a single Gram-negative Escherichia coli (E. coli) colony to 20 mL of Luria-Bertani (LB) medium and incubate at 37°C in a shaker for 12 h. Then, dilute to 3 × 10⁻⁶ with sterile PBS buffer at pH 7.4. 6 CFUmL -1 A bacterial solution was obtained. A final concentration of 1.5 μg / mL was added to the bacterial solution. -1 Cu3P (morphology as) Figure 1 As shown in (f), spherical nanoparticles with a particle size of approximately 80 nm are observed by XRD. Figure 2 (As shown in curve (f)). After treatment for 0 min and 20 min respectively, the cells were diluted with sterile PBS buffer at pH 7.4 to a colony concentration of 10. 3 CFU mL -1 The bacterial dilution was obtained. Finally, 100 μL of the bacterial dilution was evenly spread onto LB agar plates and incubated at 37°C for 24 h. The number of colonies was then counted using the plate count method. The results are as follows: Figure 4 As shown.

[0071] from Figure 4 It can be seen that the concentration is 1.5 μg / mL -1 Cu3P can achieve a bactericidal efficiency of 99.9% against Gram-negative bacteria Escherichia coli (E. coli) after 20 min.

[0072] Example 12

[0073] A single Gram-positive methicillin-resistant Staphylococcus aureus (MRSA) was transferred to 20 mL of Luria-Bertani (LB) medium and incubated for 12 h in a shaker at 37 °C. The culture was then diluted to 3 × 10⁻⁶ with sterile PBS buffer at pH 7.4. 6 CFU mL -1 A bacterial solution was obtained. A final concentration of 0.5 μg / mL was added to the bacterial solution. -1 Cu3P (morphology as) Figure 1 As shown in (f), spherical nanoparticles with a particle size of approximately 80 nm are observed by XRD. Figure 2 (As shown in curve (f)). After treatment for 0 min and 20 min respectively, the cells were diluted with PBS to a colony concentration of 10. 3 CFU mL -1 The bacterial dilution was obtained. Finally, 100 μL of the bacterial dilution was evenly spread onto LB agar plates and incubated at 37°C for 24 h. The number of colonies was then counted using the plate count method. The results are as follows: Figure 4 As shown.

[0074] from Figure 4 It can be seen that the concentration is 0.5 μg / mL -1 Cu3P achieved a bactericidal efficiency of 99.9% against methicillin-resistant Staphylococcus aureus (MRSA) in 20 minutes.

[0075] Examples 1-12 show that the size and morphology of Cu3P have little effect on the antibacterial efficiency of Cu3P.

[0076] Example 13

[0077] Cu3P (morphology as shown) Figure 1 As shown in (a), the spherical nanoparticles have a particle size of approximately 35 nm, and the XRD pattern is as follows. Figure 2 As shown in curve (a), Cu3P was dispersed in the Chagan Lake water sample, with a final concentration of 0.8 μg / mL. -1 After co-culturing at 37℃ for 5 min, 10 min, and 20 min respectively, 100 μL of water sample was evenly spread onto LB agar plates and incubated at 37℃ for 24 h. The number of colonies was then counted using the plate count method. The results are as follows: Figure 5 As shown.

[0078] from Figure 5 As can be seen, the blank (Control) culture dishes without Cu3P material treatment contained a variety of different bacterial colonies, indicating that the original lake water contained diverse bacteria. Based on gene sequencing and annotation results, the lake water contained a rich diversity of microorganisms. The 20 most abundant genera were g__GpIIa, g__Exiguobacterium, g__Bacillariophyta, g__Acinetobacter, g__unclassified_Planctomycetaceae, g__unclassified_Acidimicrobidae, g__Gemmobacter, g__unclassified_Rhizobiales, g__Phenylobacterium, g__Brevundimonas, g__unclassified_Micrococcineae, g__unclassified_Bacteria, g__Chlorophyta, g__Ilumatobacter, g__Planococcus, g__Spartobacteria_genera_incertae_sedis, g__Luteolibacter, g__unclassified_Verrucomicrobiaceae, g__Sphingomonas, and g__Paracoccus. This indicates that the lake water samples contained a large number of Gram-positive and Gram-negative bacteria, in addition to *Escherichia coli* and MRSA. The concentration of Cu3P was 0.8 μg / mL. -1 It can kill more than 99.9% of various bacteria within 20 minutes.

[0079] Example 13 shows that Cu3P material has broad-spectrum antibacterial properties.

[0080] Example 14

[0081] Single *E. coli* or MRSA bacteria were transferred to 20 mL of LB medium and incubated in a shaker at 37°C for 12 h. The culture was then diluted to 3 × 10⁻⁶ with sterile PBS buffer at pH 7.4. 6 CFUmL -1 A bacterial solution was obtained. A final concentration of 0.5 μg / mL was added to the bacterial solution. -1 1.0 μg mL -1 1.5 μg mL -1 2.0 μg mL -1 2.5 μg mL-1 5.0 μg mL -1 10.0 μg / mL -1 20.0 μg mL -1 40.0 μg mL -1 Cu3P (morphology as) Figure 1 As shown in (a), the spherical nanoparticles have a particle size of approximately 35 nm, and the XRD pattern is as follows. Figure 2 (As shown in curve (a)). After treatment for 0 min and 20 min respectively, the cells were diluted with PBS to a colony concentration of 10. 3 CFU mL -1 The bacterial dilutions were obtained. 100 μL of each bacterial dilution was evenly spread onto LB agar plates and incubated at 37°C for 24 h. The number of colonies was then counted using the plate count method. The results are as follows: Figure 6 As shown.

[0082] from Figure 6 It can be seen that the final concentrations are greater than or equal to 0.5 μg / mL. -1 and 1.5 μg mL -1 Cu3P can achieve a bactericidal efficiency of over 99.9% against Gram-positive and Gram-negative bacteria within 20 minutes.

[0083] Example 14 shows that the minimum concentration of Cu3P that exhibits significant antibacterial activity against both Gram-positive and Gram-negative bacteria is 0.5 μg / mL. -1 and 1.5 μg mL -1 Even at concentrations higher than the minimum, Cu3P exhibits significant antibacterial activity, meaning its antibacterial efficiency reaches over 99.9%. Other concentrations of Cu3P also demonstrate antibacterial activity.

[0084] Example 15

[0085] Single *E. coli* or MRSA bacteria were transferred to 20 mL of LB medium and incubated in a shaker at 37°C for 12 h. The culture was then diluted to 10 CFU / L with sterile PBS buffer at pH 7.4. -1 (Also written as 0.01 CFU / mL) -1 ), 1 CFU mL -1 100 CFU / mL -1 1×10 4 CFU mL -1 1×10 6 CFUmL -1 3×10 6 CFUmL -1 1×10 8 CFUmL -1A bacterial solution was obtained. Add mL to the bacterial solution to a final concentration of 5.0 μg. -1 Cu3P (morphology as) Figure 1 As shown in (a), the spherical nanoparticles have a particle size of approximately 35 nm, and the XRD pattern is as follows. Figure 2 (As shown in curve (a)). After treatment for 0 min and 20 min respectively, the cells were diluted with PBS to a colony concentration of 10. 3 CFU mL -1 The bacterial dilutions were obtained. 100 μL of each bacterial dilution was evenly spread onto LB agar plates and incubated at 37°C for 24 h. The number of colonies was then counted using the plate count method. The results are as follows: Figure 7 As shown.

[0086] from Figure 7 It can be seen that Cu3P material is effective at concentrations ranging from 0.01 to 1 × 10⁻⁶ per milliliter. 8 CFUmL -1 The bacteria can be sterilized with an efficiency of over 99.9% after 20 minutes of action.

[0087] Example 15 shows that the antibacterial effect of Cu3P material is not limited by the concentration of bacteria.

[0088] Example 16

[0089] Prepare 0.5 to 40.0 μg mL -1 Cu3P mother liquor (Cu3P morphology as follows) Figure 1 As shown in (a), the spherical nanoparticles have a particle size of approximately 35 nm, and the XRD pattern is as follows. Figure 2 As shown in curve (a), the solvent was distilled water, which was added dropwise to the mouse wound as a washing solution. The mouse wound was approximately 80 mm in size. 2 Incision. A blank control group was also set up, i.e., no Cu was added. 3 Group P, whose wounds healed naturally. Starting from 0 minutes, the wound area was measured every 12 hours, and the results are as follows: Figure 8 As shown, Cu3P has bactericidal and wound-healing properties.

[0090] Example 17

[0091] Cu3P powder was used as a dressing (Cu3P morphology as follows) Figure 1 As shown in (a), the spherical nanoparticles have a particle size of approximately 35 nm, and the XRD pattern is as follows. Figure 2 (As shown in curve (a)), take 0.5 to 40.0 μg cm -2 It was placed directly on a mouse wound, which was approximately 80 mm in size. 2 Incision was made. A blank control group, i.e., without Cu, was also set up. 3The wound healing naturally in group P dressing was observed. Wound area was measured every 12 hours starting from 0 min. Figure 9 The indicated dosage is 1.5 μg cm. -2 The measurement results indicate that Cu3P can be used as a medical dressing for wound sterilization and recovery.

[0092] Examples 16 and 17 illustrate Cu 3 P nanomaterials have medical antibacterial applications and can be used in liquid or solid phases.

[0093] Example 18

[0094] 100 μL of MCF-7 cells in DMEM medium were seeded into 96-well plates at a density of 5000 cells / well, with five wells per group, for a total of five groups. Cu3P (morphology as shown) was added to the five wells of each group. Figure 1 As shown in (a), the spherical nanoparticles have a particle size of approximately 35 nm, and the XRD pattern is as follows. Figure 2 As shown in curve (a), the concentration of Cu3P in each pore of the group was 0 μg / mL. -1 The concentration of Cu3P in each pore of the group was 2 μg / mL. -1 The concentration of Cu3P in each well of the group was 4 μg / mL. -1 The concentration of Cu3P in each pore of the group was 8 μg / mL. -1 The concentration of Cu3P in each pore of the group was 16 μg / mL. -1 After incubation for 24 hours, CCK-8 solution was added to each well, and incubation continued for another hour. Cell viability was then determined by measuring the absorbance at 450 nm using a microplate reader. The results are as follows: Figure 10 As shown.

[0095] from Figure 10 It can be seen that the concentration of Cu3P is 16 μg / mL. -1 At that time, it still had no significant impact on the survival rate of MCF-7 cells.

[0096] Example 19

[0097] HepG2 cells (100 μL) in DMEM medium were seeded into 96-well plates at a density of 5000 cells / well, with five wells forming a group, for a total of five groups. Cu3P (morphology as shown) was added to the five wells of each group. Figure 1 As shown in (a), the spherical nanoparticles have a particle size of approximately 35 nm, and the XRD pattern is as follows. Figure 2 As shown in curve (a), the concentration of Cu3P in each pore of the group was 0 μg / mL. -1 The concentration of Cu3P in each pore of the group was 2 μg / mL. -1The concentration of Cu3P in each well of the group was 4 μg / mL. -1 The concentration of Cu3P in each pore of the group was 8 μg / mL. -1 The concentration of Cu3P in each pore of the group was 16 μg / mL. -1 After incubation for 24 hours, CCK-8 solution was added to each well, and incubation continued for another hour. Cell viability was then determined by measuring the absorbance at 450 nm using a microplate reader. The results are as follows: Figure 10 As shown.

[0098] from Figure 10 It can be seen that the concentration of Cu3P is 16 μg / mL. -1 At that time, it had no significant effect on the survival rate of MCF-7 cells.

[0099] Examples 18 and 19 illustrate Cu 3 P material exhibits minimal toxicity to mammalian cells.

[0100] 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. Use of cuprous phosphide in the preparation of antibacterial materials; The application form of the cuprous phosphide is liquid application; The liquid application is adding cuprous phosphide into liquid containing bacteria; In the aforementioned liquid-phase application, the minimum final concentration at which cuprous phosphide achieves an inhibition rate of over 99.9% against Gram-positive bacteria is 0.5 μg / mL. -1 The minimum final concentration of cuprous phosphide to achieve an inhibition rate of over 99.9% against Gram-negative bacteria is 1.5 μg / mL. -1 ; The cuprous phosphide is spherical nanoparticle; The antibacterial material does not rely on external auxiliary conditions in use.

2. The use of the cuprous phosphide according to claim 1 in the production of an antibacterial material, characterized in that, The antibacterial time for achieving a bacteriostatic rate of 99.9% or more is 20 min or more.

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