A novel enzyme-silver antiviral and antibacterial composite preparation and a preparation method thereof

Abstract

The present application relates to the technical field of disinfectant, and specifically provides a novel enzyme-silver antiviral and antibacterial composite preparation, which comprises the following components in parts by weight: water 60-80 parts, sodium carbonate 1-3 parts, sodium chloride 1-3 parts, sodium bicarbonate 6-8 parts, citric acid 1-3 parts, sorbitol 6-8 parts, composite enzyme 3-6 parts, and nano-silver carrier 11-15 parts; the preparation is prepared from safe, non-toxic and non-irritating materials, has good storage stability and long-lasting antibacterial effect, the added composite enzyme comprises broad-spectrum nuclease, lysozyme and neutral protease, can completely eliminate viruses from the source, and the added nano-silver carrier can effectively adsorb viruses and bacteria, and has remarkable antibacterial and antiviral effect.

Description

Technical Field

[0001] This invention belongs to the technical field of disinfectants, specifically relating to a novel enzyme-silver antiviral and antibacterial compound preparation and its preparation method. Background Technology

[0002] Viruses and bacteria are ubiquitous in our environment. When our immune system is weakened, they can easily invade and cause various diseases. Therefore, hospitals, schools, and other places where people gather are frequently disinfected. However, most known disinfection methods use ingredients with strong, irritating odors, such as ether, peracetic acid, and chloroform, which can irritate the respiratory tract and even cause significant harm to the human body and central nervous system, making them unsuitable for crowded places. Enzymes, as antiviral and antibacterial components, are also used in disinfectants, but they rarely completely kill most viruses and bacteria, and their activity is easily affected by various environmental factors. Silver ions can kill almost all microorganisms, but they cannot completely eliminate some stubborn viruses, and silver ions are relatively expensive and unstable. Therefore, a novel composite disinfectant containing both enzymes and silver ions is of significant research value. Summary of the Invention

[0003] To address the above-mentioned problems, the present invention aims to provide a novel enzyme-silver antiviral and antibacterial compound preparation and its preparation method.

[0004] The technical content of this invention is as follows:

[0005] This invention provides a novel enzyme-silver antiviral and antibacterial compound formulation, comprising the following components: water, sodium carbonate, sodium chloride, sodium bicarbonate, citric acid, sorbitol, compound enzyme, and nano-silver carrier.

[0006] The compound formulation comprises the following components by weight: 60-80 parts water, 1-3 parts sodium carbonate, 1-3 parts sodium chloride, 6-8 parts sodium bicarbonate, 1-3 parts citric acid, 6-8 parts sorbitol, 3-6 parts compound enzyme, and 11-15 parts nano silver carrier.

[0007] The sodium carbonate and sodium bicarbonate have a buffering effect and also have certain bactericidal and deodorizing effects.

[0008] The citric acid has buffering, complexing and chelating effects, which enhance the bactericidal and sterilizing effects.

[0009] Sodium chloride has a solubilizing effect and a certain antibacterial effect;

[0010] The sorbitol is a protective agent that plays a protective and stabilizing role for the complex enzyme;

[0011] The complex enzyme includes a broad-spectrum nuclease, lysozyme, and neutral protease.

[0012] The complex enzyme comprises a broad-spectrum nuclease, lysozyme, and neutral protease, with a mass ratio of 2-4:200-350:150-200 for the broad-spectrum nuclease, lysozyme, and neutral protease.

[0013] The broad-spectrum nuclease is a nuclease endonuclease that can degrade nucleic acids into monophosphate nucleotides. It exhibits high efficiency in degrading any form of DNA and RNA without protein cleavage activity, and its nuclease activity reaches 1 × 10⁻⁶. 6 U / mg protein;

[0014] The neutral protease is an endonuclease that can be used for various protein hydrolysis processes.

[0015] The lysozyme is an alkaline enzyme that can hydrolyze the mucopolysaccharides in pathogenic bacteria. It can break the β-1,4 glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine in the cell wall, causing the cell wall to decompose and rupture, thereby releasing the contents and dissolving the bacteria. It can also directly bind to negatively charged viral proteins to inactivate the virus.

[0016] This invention provides a method for preparing a novel enzyme-silver antiviral and antibacterial compound formulation, comprising the following steps:

[0017] Step 1: Mix the compound enzyme, sorbitol and an appropriate amount of water, freeze dry, add the nano silver carrier and mix thoroughly for 2-3 hours to obtain the premixed solution;

[0018] The preparation method of the nano-silver carrier is as follows: sodium alginate is dissolved by stirring with an organic phosphoric acid solution of 9-13% by mass at a temperature of 60-70℃, and then 60-70 mmol / L silver nitrate solution is added. While heating and stirring, a dodecyl betaine solution of 2-3 times the volume and 10%-15% by mass is added dropwise. The mixture is then homogenized and dispersed by high-pressure microfluidic jet and allowed to stand to obtain the nano-silver carrier. The mass ratio of sodium alginate to silver nitrate solution is 5-8:2-4.

[0019] The organic phosphoric acid solution is a phytic acid solution.

[0020] Step 2: Add the remaining raw materials and water to the mixing pot and continue stirring and mixing. Then add the premixed liquid and continue to mix thoroughly. Adjust the pH to neutral to obtain the compound preparation.

[0021] The freeze-drying temperature is -15 to 20°C.

[0022] This invention provides an application of a novel enzyme-silver antiviral and antibacterial compound preparation;

[0023] The applications include, but are not limited to, the preparation of disinfectant sprays, air sterilizers, and antibacterial and antiviral wipes.

[0024] Beneficial Effects: The novel enzyme-silver antiviral and antibacterial compound preparation obtained by this invention is formulated from various safe and odorless ingredients. It is effective against most bacteria and viruses, and has a long-lasting bactericidal and disinfecting effect. The added compound enzyme is a combination of lysozyme, neutral protease and broad-spectrum nuclease. Lysozyme decomposes the bacterial cell wall and denatures the genetic material, neutral protease hydrolyzes proteins, and broad-spectrum nuclease can degrade RNA and DNA at will. The three enzymes work together to quickly achieve antibacterial and antiviral effects and can kill bacteria and viruses more thoroughly. It works synergistically with the nano-silver carrier and has a kill rate of ≥99.9% against pathogens such as Escherichia coli, Staphylococcus albus, Aspergillus niger, poliovirus, and human coronavirus.

[0025] This invention provides a method for preparing a novel enzyme-silver antiviral and antibacterial composite formulation. Sodium alginate and phytic acid solution undergo a cross-linking reaction via hydrogen bonds, giving sodium alginate antibacterial and antiviral properties. Silver nitrate solution is then added to induce a complexation reaction. The added dodecyl betaine solution disperses the bactericidal components and enhances the bactericidal effect. The phosphate groups of phytic acid have a complexing effect on metal ions. The three react with each other, improving the stability of silver ions and resulting in significant sterilization and disinfection effects. The composite enzyme is then mixed with the protective agent sorbitol and freeze-dried, which effectively maintains the enzyme's activity. Further mixing allows the freeze-dried composite enzyme to be encapsulated by a nano-silver carrier, further stabilizing the enzyme through slow release. The resulting formulation has a stability period of ≥90 days at room temperature and a half-life of ≥30 days for the bio-enzyme activity. Simultaneously, the composite enzyme and silver ions work together to target bacteria and viruses adsorbed by the nano-silver carrier, making the entire sterilization and virus eradication process rapid and efficient.

[0026] This invention provides an application of a novel enzyme-silver antiviral and antibacterial compound preparation. This compound preparation is safe and harmless, leaves no residue after use, and can be applied to various disinfection needs in daily life, such as air disinfection, human skin disinfection, and object disinfection. It can also be used in the research and development of related disinfection equipment. Detailed Implementation

[0027] The present invention will be further described in detail below through specific implementation examples. It should be understood that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of protection of the present invention. After reading the present invention, any modifications of the present invention in various equivalent forms by those skilled in the art fall within the scope of the appended claims.

[0028] Unless otherwise specified, all raw materials and reagents used in this invention are from the conventional market.

[0029] Example 1

[0030] A novel enzyme-silver antiviral and antibacterial compound formulation comprises the following components by weight: 70 parts water, 2 parts sodium carbonate, 2 parts sodium chloride, 7 parts sodium bicarbonate, 2 parts citric acid, 7 parts sorbitol, 4 parts compound enzyme, and 13 parts nano silver carrier.

[0031] The complex enzyme comprises a broad-spectrum nuclease, lysozyme, and neutral protease, with a mass ratio of 3:275:175.

[0032] The preparation of the novel enzyme-silver antiviral and antibacterial compound formulation includes the following steps:

[0033] Step 1: Mix the compound enzyme, sorbitol and an appropriate amount of water, freeze-dry at -17℃, add the nano silver carrier and mix thoroughly for 3 hours to obtain the premixed solution;

[0034] The preparation method of the nano-silver carrier is as follows: sodium alginate is dissolved in 11% phytic acid solution at 65°C by stirring, then 65 mmol / L silver nitrate solution is added. While heating and stirring, dodecyl betaine solution with a volume ratio of 2 times and a mass concentration of 13% is added dropwise. The mixture is then homogenized and dispersed using high-pressure microfluidic jet. After standing, the nano-silver carrier is obtained. The mass ratio of sodium alginate to silver nitrate solution is 6:3.

[0035] Step 2: Add the remaining raw materials and water to the mixing pot and continue stirring and mixing. Then add the premixed liquid and continue to mix thoroughly. Adjust the pH to neutral to obtain the compound preparation.

[0036] Example 2

[0037] A novel enzyme-silver antiviral and antibacterial compound formulation comprises the following components by weight: 60 parts water, 1 part sodium carbonate, 1 part sodium chloride, 6 parts sodium bicarbonate, 1 part citric acid, 6 parts sorbitol, 3 parts compound enzyme, and 11 parts nano silver carrier.

[0038] The complex enzyme comprises a broad-spectrum nuclease, lysozyme, and neutral protease, with a mass ratio of 2:200:150 for the broad-spectrum nuclease, lysozyme, and neutral protease.

[0039] The preparation of the novel enzyme-silver antiviral and antibacterial compound formulation includes the following steps:

[0040] Step 1: Mix the compound enzyme, sorbitol and an appropriate amount of water, freeze-dry at -15℃, add the nano silver carrier and mix thoroughly for 2 hours to obtain the premixed solution;

[0041] The preparation method of the nano-silver carrier is as follows: sodium alginate is dissolved in 9% phytic acid solution at 60°C by stirring, then 60 mmol / L silver nitrate solution is added, and while heating and stirring, a 10% dodecyl betaine solution with a volume ratio of 2 is added dropwise. The mixture is then homogenized and dispersed using a high-pressure microfluidic jet, and allowed to stand to obtain the nano-silver carrier. The mass ratio of sodium alginate to silver nitrate solution is 5:2.

[0042] Step 2: Add the remaining raw materials and water to the mixing pot and continue stirring and mixing. Then add the premixed liquid and continue to mix thoroughly. Adjust the pH to neutral to obtain the compound preparation.

[0043] Example 3

[0044] A novel enzyme-silver antiviral and antibacterial compound preparation comprises the following components by weight: 80 parts water, 3 parts sodium carbonate, 3 parts sodium chloride, 8 parts sodium bicarbonate, 3 parts citric acid, 8 parts sorbitol, 6 parts compound enzyme, and 15 parts nano silver carrier.

[0045] The complex enzyme comprises a broad-spectrum nuclease, lysozyme, and neutral protease, with a mass ratio of 4:350:200 for the broad-spectrum nuclease, lysozyme, and neutral protease.

[0046] The preparation of the novel enzyme-silver antiviral and antibacterial compound formulation includes the following steps:

[0047] Step 1: Mix the compound enzyme, sorbitol and an appropriate amount of water, freeze-dry at -20℃, add the nano silver carrier and mix thoroughly for 3 hours to obtain the premixed solution;

[0048] The preparation method of the nano-silver carrier is as follows: sodium alginate is dissolved in phytic acid solution with a mass fraction of 13% at 70°C by stirring, and then 70 mmol / L silver nitrate solution is added. While heating and stirring, dodecyl betaine solution with a volume ratio of 3 times and a mass concentration of 15% is added dropwise. The mixture is then homogenized and dispersed using high-pressure microfluidic jet. After standing, the nano-silver carrier is obtained. The mass ratio of sodium alginate to silver nitrate solution is 8:4.

[0049] Step 2: Add the remaining raw materials and water to the mixing pot and continue stirring and mixing. Then add the premixed liquid and continue to mix thoroughly. Adjust the pH to neutral to obtain the compound preparation.

[0050] Comparative Example 1

[0051] The difference between Comparative Example 1 and Example 1 is that no broad-spectrum nuclease was added to the complex enzyme in Comparative Example 1, and its amount was replaced by an average increase of the other two enzymes, while other reaction conditions remained unchanged.

[0052] Comparative Example 2

[0053] The difference between Comparative Example 2 and Example 1 is that lysozyme is not added to the complex enzyme in Comparative Example 2, but its amount is replaced by an average increase of the other two enzymes, while other reaction conditions remain unchanged.

[0054] Comparative Example 3

[0055] The difference between Comparative Example 3 and Example 1 is that no phytic acid solution was added to the silver nanoparticle carrier in Comparative Example 3, while other reaction conditions remained unchanged.

[0056] Comparative Example 4

[0057] The difference between Comparative Example 4 and Example 1 is that Comparative Example 4 directly adds silver nitrate solution instead of silver nanoparticle carrier, while other reaction conditions remain unchanged.

[0058] Efficacy verification

[0059] I. Sterilization effect test

[0060] According to the "Disinfection Technical Specifications" (2002 edition), for 20m 3 The test chamber was sterilized, and then the bacterial suspension was diluted and sprayed into the entire test chamber. 4L of the disinfectant preparations prepared in the examples and comparative examples were sprayed into the test chamber for disinfection for 1 hour. Sampling was carried out using a QSP-B-A026 six-stage airborne microbial sampler, and then incubated in plates for 48 hours. The bacterial content before and after disinfection was detected to obtain the sterilization rate of the disinfectant preparation. The experiment was repeated 3 times, and the average value was taken. At the same time, each sample was placed under the same conditions for 3 months, and the experiment was repeated to test its bactericidal effect. The results are shown in Table 1.

[0061] Table 1

[0062]

[0063] As shown in Table 1, the compound preparation prepared by this invention has significant sterilization effects on Escherichia coli, Staphylococcus albus and Aspergillus niger, with a sterilization rate of over 99%, meeting the qualified standards. Its effective bactericidal active ingredients have high stability, and under normal storage conditions, its bactericidal effect lasts for more than 3 months without any significant weakening.

[0064] II. Disinfection Efficacy Test

[0065] The test reference was based on the "Disinfection Technical Specifications" (2002 edition). The test groups of the example and comparative examples were tested at 0 days and 30 days, respectively. The test results are shown in Table 2.

[0066] Table 2

[0067]

[0068]

[0069] As shown in Table 2, the present invention has a high elimination rate for common infectious viruses, with elimination rates of ≥99.9% for poliovirus, human coronavirus, and influenza A virus. Combined with Table 1, it can be seen that the compound enzyme added in the present invention consists of lysozyme, neutral protease, and broad-spectrum nuclease, which have synergistic effects. Reducing the types of enzymes will affect the bactericidal effect. At the same time, the nano-silver carrier is prepared by silver nitrate solution, sodium alginate, and phytic acid. Compared with nano-silver carriers prepared by single nano-silver or without phytic acid, it has more stable and powerful bactericidal and disinfection effects.

[0070] This invention utilizes the combined action of three enzymes to effectively hydrolyze the viral protein coat and nucleotide chains, thereby inactivating the virus. Freeze-drying protects and maintains enzyme activity. Furthermore, nano-silver, phytic acid, and sodium alginate cross-link to form a nano-silver carrier, which possesses potent bactericidal and disinfecting properties. Dodecyl betaine solution acts as a dispersant and also adsorbs the complex enzyme, maintaining its stability. When sprayed onto object surfaces, it adsorbs bacteria and viruses. The combined action of the silver carrier, enzymes, and other components achieves long-lasting and highly effective antibacterial properties.

Claims

1. A novel enzyme-silver antiviral and antibacterial compound preparation, characterized in that, It contains the following components: water, sodium carbonate, sodium chloride, sodium bicarbonate, citric acid, sorbitol, complex enzymes, and nano-silver carriers; By weight, the compound preparation contains 60-80 parts water, 1-3 parts sodium carbonate, 1-3 parts sodium chloride, 6-8 parts sodium bicarbonate, 1-3 parts citric acid, 6-8 parts sorbitol, 3-6 parts compound enzyme, and 11-15 parts nano silver carrier. The complex enzyme comprises a broad-spectrum nuclease, lysozyme, and neutral protease, with a mass ratio of 2-4: 200-350: 150-200. The preparation method of the nano-silver carrier is as follows: sodium alginate is dissolved in an organic phosphoric acid solution with a mass fraction of 9-13% at a temperature of 60-70℃ by stirring, and then a silver nitrate solution with a mass concentration of 60-70 mmol / L is added. While heating and stirring, a dodecyl betaine solution with a volume multiple of 2-3 times and a mass concentration of 10%-15% is added dropwise. The mixture is then homogenized and dispersed using a high-pressure microfluidic jet and allowed to stand to obtain the nano-silver carrier. The mass ratio of sodium alginate to silver nitrate solution is 5-8:2-4.

2. The novel enzyme-silver antiviral and antibacterial compound preparation according to claim 1, characterized in that, The organic phosphoric acid solution is a phytic acid solution.

3. The preparation method of a novel enzyme-silver antiviral and antibacterial compound preparation according to claim 1 or 2, characterized in that, Includes the following steps: Step 1: Mix the compound enzyme, sorbitol and water, freeze-dry, add the silver nanoparticle carrier and mix thoroughly for 2-3 hours to obtain the premixed solution; Step 2: Add the remaining raw materials and water to the mixing pot and continue stirring and mixing. Then add the premixed liquid and continue to mix thoroughly. Adjust the pH to neutral to obtain the compound preparation.

4. The preparation method of a novel enzyme-silver antiviral and antibacterial compound preparation according to claim 3, characterized in that, The freeze-drying temperature is -15 to -20°C.

5. The application of the novel enzyme-silver antiviral and antibacterial compound preparation according to claim 1 or 2, characterized in that, The applications include, but are not limited to, the preparation of disinfectant sprays, air sterilizers, and antibacterial and antiviral wipes.

Images