Low-temperature stable antibacterial gel nanomaterial, and preparation method and application thereof

Abstract

The present application relates to the technical field of disinfectant, in particular to a low-temperature stable antibacterial gel nanomaterial and a preparation method and application thereof.The disinfectant comprises a low-temperature resistant disinfecting hydrogel and a calcium chloride aqueous solution, and the low-temperature resistant disinfecting hydrogel is dispersed in the calcium chloride aqueous solution in the form of microspheres.The low-temperature resistant disinfecting hydrogel is prepared from raw materials including quaternary ammonium cation chloride, acrylic acid, calcium chloride and water, and the molar ratio of the quaternary ammonium cation chloride, the acrylic acid and the calcium chloride is 1:(1-3):(1-3).The disinfectant of the present application can make the low-temperature resistant disinfecting hydrogel dispersed in the calcium chloride aqueous solution in the form of microspheres, the low-temperature resistant disinfecting hydrogel in the form of microspheres has a large specific surface area and good dispersibility, can be in contact with the surface of the object to be disinfected more uniformly, and can improve the disinfection efficiency, and this unique dispersion system can make the disinfectant still have good fluidity and bactericidal effect in a low-temperature environment.

Description

Technical Field

[0001] This invention relates to the field of disinfectant technology, and in particular to a low-temperature stable antibacterial gel nanomaterial, its preparation method, and its application. Background Technology

[0002] In low-temperature environments, sterilization and disinfection are required in various scenarios. In cold chain transportation and storage, such as food and pharmaceutical cold chains, packaging, equipment, and storage spaces must be disinfected to prevent microbial contamination. In medical institutions, low-temperature operating rooms and cryopreservation equipment must ensure the hygiene and safety of surgical instruments, equipment, and preserved samples. In densely populated areas such as public places and winter sports venues in cold northern regions, low-temperature disinfection of public facilities and equipment is necessary to reduce the risk of pathogen transmission. In industrial production, low-temperature food processing workshops and low-temperature equipment in chemical enterprises also require disinfection to ensure product quality and equipment safety. These disinfection efforts in low-temperature environments are crucial for ensuring public health safety, maintaining product quality, and ensuring stable equipment operation.

[0003] There are many types of commercially available disinfectants, including quaternary ammonium salt disinfectants, chlorine-containing disinfectants, and peroxide disinfectants. However, these conventional disinfectants usually solidify at low temperatures, which greatly reduces their bactericidal ability and makes them unable to achieve a bactericidal effect at low temperatures. Summary of the Invention

[0004] This invention provides a low-temperature stable antibacterial gel nanomaterial, its preparation method, and its application, in order to solve the problem that existing disinfectants will become less effective or ineffective when used in low-temperature environments.

[0005] According to a first aspect of the present invention, the present invention provides a low-temperature stable antibacterial gel nanomaterial, comprising a low-temperature resistant disinfection hydrogel and a calcium chloride aqueous solution, wherein the low-temperature resistant disinfection hydrogel is dispersed in the calcium chloride aqueous solution in the form of microspheres; the low-temperature resistant disinfection hydrogel is prepared from raw materials comprising quaternary ammonium cationic chloride, acrylic acid, calcium chloride and water, wherein the molar ratio of quaternary ammonium cationic chloride, acrylic acid and calcium chloride is 1∶(1-3)∶(1-3).

[0006] This invention discloses a low-temperature stable antibacterial gel nanomaterial. A low-temperature resistant disinfectant hydrogel is dispersed in microspheres in a calcium chloride aqueous solution. The microspheres of the low-temperature resistant disinfectant hydrogel have a large specific surface area and good dispersibility, enabling more uniform contact with the surface of the object to be disinfected, thus improving disinfection efficiency. This unique dispersion system allows the disinfectant to maintain good fluidity and bactericidal effect even at low temperatures, solving the problem of traditional disinfectants solidifying and becoming ineffective at low temperatures. The low-temperature resistant disinfectant hydrogel is prepared from raw materials such as quaternary ammonium cationic chloride, acrylic acid, calcium chloride, and water. These components synergistically enhance the bactericidal ability of the disinfectant while ensuring its stability at low temperatures.

[0007] Preferably, the molar ratio of quaternary ammonium cationic chloride, acrylic acid, and calcium chloride is 1:(1.5-2.5):(1.5-2.5). More preferably, the molar ratio of quaternary ammonium cationic chloride, acrylic acid, and calcium chloride is 1:2:2.

[0008] Furthermore, the volume ratio of acrylic acid to water is (0.5-1.5):1. This ratio range has been optimized to ensure the formation of a stable, low-eutectic solvent during the preparation process, thereby producing a high-performance, low-temperature resistant disinfectant hydrogel. If the ratio exceeds this range, the properties of the mixed solution may be unstable, affecting the subsequent preparation results and the performance of the disinfectant.

[0009] Furthermore, the volume ratio of acrylic acid to water is (0.8-1.2):1. In some specific embodiments, the volume ratio of acrylic acid to water is 1:1.

[0010] Furthermore, the calcium chloride aqueous solution contains 10-30% calcium chloride by mass. This concentration range provides a suitable dispersion environment for the low-temperature resistant disinfectant hydrogel, allowing the microspheres to be stably dispersed within it, preventing gel particle aggregation or precipitation, thereby ensuring the stability of the disinfectant during storage and use. In addition, calcium chloride itself has a certain bactericidal effect, and its presence within this concentration range can synergistically enhance the bactericidal ability of the disinfectant and improve its effectiveness against various pathogens.

[0011] Furthermore, the calcium chloride aqueous solution contains 15-25% calcium chloride by mass, preferably 18-22%. In some specific embodiments, the calcium chloride aqueous solution contains 20% calcium chloride by mass.

[0012] Furthermore, the quaternary ammonium cationic chloride is selected from one or both of choline chloride and benzalkonium chloride. Both of these compounds are effective bactericidal components with good bactericidal properties and biocompatibility. Choline chloride and benzalkonium chloride differ in structure and bactericidal mechanism; using them alone or in combination can achieve synergistic bactericidal effects, improve the disinfectant's ability to kill different types of pathogens, and enhance the broad-spectrum bactericidal performance of the disinfectant.

[0013] Furthermore, the low-temperature resistant disinfectant hydrogel has a particle size of 80-90 nm. This particle size range allows the gel particles to be uniformly dispersed in the calcium chloride aqueous solution, avoiding aggregation or precipitation caused by excessively large particles, thus ensuring the stability and uniformity of the disinfectant. In addition, this particle size of the low-temperature resistant disinfectant hydrogel has a large specific surface area, enabling the disinfectant to make more thorough contact with the surface of the object to be disinfected, improving disinfection efficiency, reducing the amount of disinfectant used, and also facilitating the rapid diffusion and action of the disinfectant in low-temperature environments.

[0014] Furthermore, the temperature of the low temperature is ≤-20℃.

[0015] According to a second aspect of the present invention, the present invention also provides a method for preparing the above-mentioned low-temperature stable antibacterial gel nanomaterial, comprising the following steps:

[0016] Step (1): Mix acrylic acid, water, quaternary ammonium cationic chloride and calcium chloride to obtain a mixed solution;

[0017] Step (2): Heat the mixed solution in a water bath at 50-70°C for 10-30 minutes to obtain a clear and transparent low eutectic solvent;

[0018] Step (3): Add the eutectic solvent into the mold and irradiate with ultraviolet light to obtain a transparent low-temperature sterilization resistant hydrogel;

[0019] Step (4): The low-temperature resistant disinfection hydrogel is added to a calcium chloride aqueous solution for ultrasonic crushing to obtain a low-temperature stable antibacterial gel nanomaterial.

[0020] The present invention discloses a method for preparing low-temperature stable antibacterial gel nanomaterials, comprising steps such as mixing raw materials, heat treatment, ultraviolet irradiation, and ultrasonic disruption. This method ensures the operability and repeatability of the disinfectant preparation process, facilitating industrial production and practical application. By strictly controlling the conditions of each step, such as temperature, time, and ultraviolet irradiation parameters, the prepared disinfectant can be guaranteed to have stable performance and good quality, meeting the disinfection requirements for use in low-temperature environments.

[0021] Furthermore, in step (3), the ultraviolet lamp used for ultraviolet irradiation has a wavelength of 365nm and a power of 800-900mW / cm².2 The irradiation time is 20-40 seconds.

[0022] By optimizing the ultraviolet irradiation conditions, the eutectic solvent can undergo a rapid and uniform polymerization reaction under ultraviolet light to form a transparent, low-temperature resistant sterilization hydrogel, while avoiding side reactions or degradation of gel performance caused by excessive irradiation.

[0023] Furthermore, in step (3), the eutectic solvent is added into the mold until the liquid level is flush with the mold surface. This operational detail of adding the eutectic solvent into the mold until the liquid level is flush with the mold surface ensures the molding quality of the gel, giving it a uniform thickness and good appearance, and further improving the effectiveness and stability of the disinfectant.

[0024] Preferably, the mold is a silicon mold.

[0025] Furthermore, in step (4), the shear rate of the ultrasonic disruption is 10,000-30,000 rpm, and the time is 30-90 s. Suitable ultrasonic disruption parameters can effectively break down the low-temperature resistant disinfectant hydrogel into smaller particles, making it more uniformly dispersed in the calcium chloride aqueous solution, thereby further improving the stability and disinfection efficiency of the disinfectant.

[0026] Furthermore, in step (4), the shear rate of ultrasonic fragmentation is 15000-25000 rpm, and the time is 50-70 s. In some specific embodiments, in step (4), the shear rate of ultrasonic fragmentation is 20000 rpm, and the time is 1 min.

[0027] According to a third aspect of the present invention, the present invention also provides the application of the above-described low-temperature stable antibacterial gel nanomaterial or the low-temperature stable antibacterial gel nanomaterial prepared by the above-described method for preparing disinfectants for killing African swine fever virus and / or pseudorabies virus and / or avian influenza H1N1.

[0028] The disinfectant of this invention can be used to prepare disinfectants for killing African swine fever virus, pseudorabies virus, and avian influenza H1N1 virus. These three viruses are pathogens of significant public health importance. Applying this disinfectant to kill these viruses can effectively reduce the risk of virus transmission and ensure public health safety. Considering the characteristics of these viruses, this disinfectant maintains good bactericidal effects even at low temperatures (≤-20℃), meeting the needs for virus killing in special scenarios such as cold chain transportation and low-temperature storage, and filling a gap in existing technologies in the field of low-temperature disinfection.

[0029] Furthermore, in the disinfectant for killing African swine fever virus and / or pseudorabies virus and / or avian influenza H1N1, the mass concentration of the low-temperature stable antibacterial gel nanomaterial is 0.025-0.05%. This concentration range has been experimentally verified to ensure that the disinfectant effectively kills the virus while avoiding increased costs and potential environmental impacts caused by excessively high concentrations.

[0030] This invention provides a low-temperature stable antibacterial gel nanomaterial by dispersing a low-temperature resistant disinfectant hydrogel in microspheres in a calcium chloride aqueous solution. The microsphere-shaped low-temperature resistant disinfectant hydrogel has a large specific surface area and good dispersibility, enabling more uniform contact with the surface of the object to be disinfected, thus improving disinfection efficiency. This unique dispersion system allows the disinfectant to maintain good fluidity and bactericidal effect at low temperatures, solving the problem of traditional disinfectants solidifying and becoming ineffective at low temperatures. The low-temperature resistant disinfectant hydrogel is prepared from raw materials such as quaternary ammonium cationic chloride, acrylic acid, calcium chloride, and water. These components synergistically enhance the bactericidal ability of the disinfectant at a specific molar ratio, while ensuring its stability at low temperatures. Attached Figure Description

[0031] To more clearly illustrate the technical solutions in this invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0032] Figure 1 Transmission electron microscopy image of a low-temperature stable antibacterial gel nanomaterial provided in Embodiment 1 of the present invention.

[0033] Figure 2 This is one of the images showing the bactericidal effect of the disinfectant obtained in Example 1 of the present invention on Escherichia coli.

[0034] Figure 3 This is the second diagram showing the bactericidal effect of the disinfectant obtained in Example 1 of the present invention on Escherichia coli. Detailed Implementation

[0035] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0036] The sources of the raw materials used in the following examples and comparative examples are as follows:

[0037] Acrylic acid (AA), purity: AR, >99% (GC), manufactured by McLean.

[0038] Choline chloride (ChCl), 99% purity, manufactured by Inokai.

[0039] Benzalkonium chloride (BC), 99% purity, manufactured by McLean.

[0040] Calcium chloride, 99% purity, manufactured by Solarbio.

[0041] Sodium chloride, 99% purity, manufactured by Solarbio.

[0042] Example 1

[0043] This embodiment provides a low-temperature stable antibacterial gel nanomaterial, comprising a low-temperature resistant disinfection hydrogel and a calcium chloride aqueous solution. The low-temperature resistant disinfection hydrogel is dispersed in microspheres in the calcium chloride aqueous solution. The low-temperature resistant disinfection hydrogel is prepared from raw materials including quaternary ammonium cationic chloride, acrylic acid, calcium chloride, and water, wherein the molar ratio of quaternary ammonium cationic chloride, acrylic acid, and calcium chloride is 1:2:2. The mass concentration of the calcium chloride aqueous solution is 20%.

[0044] The preparation method of this disinfectant includes the following steps:

[0045] 0.072 mol of acrylic acid and water were mixed evenly at a volume ratio of 1:1. Then, 0.036 mol of choline chloride and 0.072 mol of calcium chloride were added to a glass container and mixed evenly to prepare a mixed solution.

[0046] The mixed solution was placed in a constant temperature heating magnetic stirrer and heated in a water bath at 60°C for 20 minutes to obtain a clear, transparent, homogeneous and stable solution.

[0047] Add an appropriate amount of solution to the silicone mold until the liquid level is flush with the mold surface, and then expose it to a 365 nm ultraviolet lamp (850 mW / cm²). 2 A transparent, low-temperature resistant disinfectant hydrogel can be prepared by irradiation for 60 seconds.

[0048] Low-temperature resistant sterilization hydrogel was added to a beaker containing 100 mL of 20 wt% calcium chloride aqueous solution and subjected to ultrasonic disruption to obtain low-temperature stable antibacterial gel nanomaterials. Among these, the low-temperature stable antibacterial gel nanomaterials include... Figure 1 As shown, the low-temperature resistant disinfection hydrogel is dispersed in calcium chloride aqueous solution in the form of nanospheres with a particle size of 86.2±2.4nm.

[0049] Comparative Example 1

[0050] This comparative example provides a disinfectant whose preparation method differs from that of Example 1 only in that sodium chloride is used instead of calcium chloride.

[0051] Comparative Example 2

[0052] This comparative example provides a disinfectant whose preparation method differs from that of Example 1 only in that sodium hypochlorite is used instead of quaternary ammonium cationic chloride.

[0053] Example 2

[0054] The disinfectant obtained in Example 1 was subjected to a bactericidal experiment. The experimental method was as follows: the disinfectant in Example 1 was diluted with water to a mass concentration of 0.002%, and Escherichia coli (10) bacteria were added. 6 CFU precipitate was mixed with 0.002% disinfectant at -20°C for 1 min and 5 min respectively; then centrifuged at 6000 rpm for 5 min, resuspended in 100 μL LB broth, spread on LB solid medium, and incubated overnight. Images were taken and the samples were counted for analysis. Figure 2 As shown, the disinfectant of the present invention can kill all Escherichia coli within 1 minute when the mass concentration is 0.002%.

[0055] Example 3

[0056] The disinfectant obtained in Example 1 was used to conduct a metal surface sterilization experiment. The experimental method was as follows: (1) The disinfectant in Example 1 was diluted with water to 0.002%, and 1 ml of the 0.002% disinfectant was spread on the metal surface and placed at -20°C; (2) 100 μl of Escherichia coli (10 μL) was added to the metal surface on the 7th, 14th, 21st and 28th days respectively. 6 (3) After 1 min, take a sample and place it in a neutralizing agent (1 L PBS plus 1 g sodium thiosulfate + 10 g Tween 80 + 5 g lecithin); (4) then centrifuge at 6000 rpm for 5 min, resuspend in 100 μL LB culture medium, spread on a solid culture medium, and incubate overnight. Take pictures and count the results for analysis. Figure 3 As shown, the disinfectant of the present invention can stably kill all Escherichia coli within 4 weeks at a mass concentration of 0.002%.

[0057] Example 4

[0058] The disinfectant obtained in Example 1 was subjected to a virus sterilization experiment. The experimental method was as follows:

[0059] (1) Take 1 ml of virus (10CT <) suspension and drop it vertically to the center of the carrier (placed at -20℃). Use tweezers to hold the edge of the filter paper to absorb and spread it.

[0060] (2) Using a calibration spray bottle (output 0.8 mL / s), spray the surface at a uniform speed in a Z-shaped trajectory from a distance of 30 cm, using 10 mL, and let it stand until the specified time point.

[0061] (3) Three sampling areas were selected using the nine-square grid method. Samples were taken at 10 min, 20 min, and 30 min respectively. After sampling, the samples were placed in a neutralizing agent (1L PBS plus 1g sodium thiosulfate + 10g Tween 80 + 5g lecithin).

[0062] (4) Viral nucleic acid was extracted for RT-PCR reaction, and the CT value was calculated. The experimental results for African swine fever virus, pseudorabies virus and avian influenza H1N1 virus are shown in Tables 1, 2 and 3 below.

[0063] Table 1

[0064]

[0065] As shown in Table 1, in the RT-PCR experiment, the disinfectant of the present invention was diluted with water to prepare disinfectant solutions with a mass concentration of 0.025% and 0.05%. The disinfectant of the present invention completely degraded the nucleic acid of African swine fever virus at a mass concentration of 0.05%, thus having a good effect on the prevention and control of African swine fever.

[0066] Table 2

[0067]

[0068] As can be seen from the experimental results in Table 2, in the RT-PCR experiment, the disinfectant of the present invention completely degraded the nucleic acid of pseudorabies virus at 0.025%.

[0069] Table 3

[0070]

[0071] As can be seen from the results in Table 3, in the RT-PCR experiment, the disinfectant of the present invention completely degraded the nucleic acid of avian influenza H1N1 virus at 0.025%.

[0072] The effects of Example 1, Comparative Example 1, and Comparative Example 2 were compared in the following disinfection experiment. The experimental method was as follows:

[0073] (1) Take 1 ml of virus (10CT <) suspension and drop it vertically to the center of the carrier (placed at -20℃). Use tweezers to hold the edge of the filter paper to absorb and spread it.

[0074] (2) Using a calibrated spray bottle (output volume 0.8 mL / s), spray disinfectant (mass concentration of 0.05%) at a uniform speed in a Z-shaped trajectory at a distance of 30 cm from the surface, using 10 mL, and let it stand until the specified time point.

[0075] (3) Three sampling areas were selected using the nine-square grid method. Samples were taken at 10 min, 20 min, and 30 min respectively. After sampling, the samples were placed in a neutralizing agent (1L PBS plus 1g sodium thiosulfate + 10g Tween 80 + 5g lecithin).

[0076] (4) Viral nucleic acid was extracted for RT-PCR reaction, and the CT value was calculated. The experimental results for African swine fever virus, pseudorabies virus and avian influenza H1N1 virus are shown in Tables 4, 5 and 6 below.

[0077] Table 4: The effectiveness of disinfectants in eliminating African swine fever virus

[0078]

[0079] Table 5: The effectiveness of disinfectants in eliminating pseudorabies virus.

[0080]

[0081] Table 6: The effectiveness of disinfectants in eliminating avian influenza H1N1 virus

[0082]

[0083] The experimental data in Tables 4-6 show that the disinfectant in Example 1 has a significant inactivation effect on African swine fever virus, pseudorabies virus, and avian influenza H1N1 virus at a low temperature of -20℃. Its CT values ​​were undetectable (NA) at 10 min, 20 min, and 30 min, indicating that the viral nucleic acid was completely degraded. In contrast, the disinfectants in Comparative Example 1 (using sodium chloride instead of calcium chloride) and Comparative Example 2 (using sodium hypochlorite instead of quaternary ammonium cationic chloride) were less effective; their CT values ​​still showed the presence of viral nucleic acid at the same time points, indicating that the virus was not completely inactivated. The CT values ​​of the control group showed that viral nucleic acid could be detected normally without the use of disinfectant, further demonstrating the high efficiency of the disinfectant in Example 1.

[0084] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A cryo-stable antibacterial gel nanomaterial, characterized in that, The low-temperature resistant disinfection hydrogel is dispersed in the calcium chloride aqueous solution in the form of microspheres; the low-temperature resistant disinfection hydrogel is prepared from raw materials including quaternary ammonium cation chloride, acrylic acid, calcium chloride and water, wherein the molar ratio of the quaternary ammonium cation chloride, the acrylic acid and the calcium chloride in the low-temperature resistant disinfection hydrogel is 1: (1-3): (1-3).

2. The cryostabilized antimicrobial gel nanomaterial of claim 1, wherein, The volume ratio of the acrylic acid to the water is (0.5-1.5): 1; And / or, the mass percentage of the calcium chloride in the calcium chloride aqueous solution is 10-30%.

3. The cryostabilized antimicrobial gel nanomaterial of claim 1, wherein, The quaternary ammonium cation chloride is selected from one or both of choline chloride or benzalkonium chloride.

4. The cryostabilized antimicrobial gel nanomaterial of claim 1, wherein, The particle size of the low-temperature resistant disinfection hydrogel is 80-90 nm.

5. The cryo-stabilized antimicrobial gel nanomaterial of claim 1, wherein, The temperature of the low temperature is ≤-20℃.

6. The process for the preparation of cryostabilized antimicrobial gel nanomaterial according to any one of claims 1 to 5, characterized in that, The method comprises the following steps: Step (1): mixing acrylic acid, water, quaternary ammonium cation chloride and calcium chloride to obtain a mixed solution; Step (2): heating the mixed solution in a water bath at 50-70℃ for 10-30 min to obtain a clear and transparent low eutectic solvent; Step (3): adding the low eutectic solvent into a mold and performing ultraviolet irradiation to obtain a transparent low-temperature resistant disinfection hydrogel; Step (4): adding the low-temperature resistant disinfection hydrogel into a calcium chloride aqueous solution and performing ultrasonic crushing to obtain a low-temperature stable antibacterial gel nanomaterial.

7. The process for the preparation of cryostabilized antimicrobial gel nanomaterial according to claim 6, characterized in that, In the step (3), the ultraviolet lamp used in the ultraviolet irradiation has a wavelength of 365 nm, a power of 800-900 mW / cm 2 , and an irradiation time of 20-40 s. And / or, in step (3), the low eutectic solvent is added into the mold until the liquid surface is flush with the surface of the mold.

8. The process for the preparation of cryostabilized antimicrobial gel nanomaterial according to claim 6, characterized in that, In step (4), the shear rate of the ultrasonic crushing is 10000-30000 rpm, and the time is 30-90 s.

9. Use of the low-temperature stable antibacterial gel nanomaterial of any one of claims 1-5 or the low-temperature stable antibacterial gel nanomaterial prepared by the method of any one of claims 6-8 in the preparation of a disinfectant for killing African swine fever virus and / or pseudorabies virus and / or avian influenza H1N1.

10. Use according to claim 9, characterized in that, In the disinfectant for killing African swine fever virus and / or pseudorabies virus and / or avian influenza H1N1, the mass concentration of the low-temperature stable antibacterial gel nanomaterial is 0.025-0.05%.

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