Small-molecule protein environment air disinfecting and deodorizing preparation and application thereof

Abstract

The present application relates to the technical field of environmental purification and odor removal, and discloses a small-molecule protein environmental air disinfection and odor removal preparation and application thereof.The preparation is made of core protein components, sodium citrate, sorbitol, an anti-browning agent, L-sodium ascorbate, riboflavin, cyclodextrin or cyclodextrin derivatives, a surfactant, a preservative and deionized water, wherein the core protein components are enzymatic products with a molecular weight less than or equal to 20 kDa, and the preparation is prepared by establishing an anti-browning buffer environment, dissolving functional components, slowly adding the core protein components to make them disperse and dissolve, and finally adjusting the pH value and setting the volume.The core protein components with a molecular weight less than or equal to 20 kDa have more active sites, and can enhance the capture and combination capacity with indoor air chemical pollutants, and can act on bacterial cell structures or virus activity, so as to realize the multifunctional air purification effect of pollutant removal and biological disinfection and inactivation.

Description

Technical Field

[0001] This invention relates to the field of environmental purification and odor elimination technology, specifically to a small molecule protein-based environmental air disinfection and deodorization preparation and its application. Background Technology

[0002] With socio-economic development and improved living standards, people's demands for indoor air quality are increasing. Indoor pollutants come from a wide range of sources, mainly including chemical pollutants (such as formaldehyde, ammonia, benzene, and total volatile organic compounds) and biological pollutants (such as bacteria, mold, and viruses). These pollutants not only produce unpleasant odors but also pose a threat to human health. Currently, the main technologies for treating these pollutants include physical adsorption, chemical oxidation, and biological purification technologies.

[0003] Existing air purification technologies have their limitations in practical applications. Physical adsorption technology (represented by activated carbon) mainly utilizes the porous structure of materials for adsorption, but this method only physically captures pollutants and cannot degrade them. Furthermore, the adsorption materials easily reach adsorption saturation and are difficult to regenerate. Simultaneously, it has low removal efficiency for polar small molecules such as formaldehyde and amines. Chemical oxidation technology (such as using ozone or hydrogen peroxide) reacts rapidly, but the oxidants used are usually highly corrosive and irritating, causing harm to equipment and human health while removing odors. Moreover, it lacks the ability to specifically degrade complex odor molecules, easily leading to secondary pollution.

[0004] Bioremediation technologies, particularly those using enzymes or protein-based formulations, have garnered attention due to their natural origin and biodegradability. These formulations utilize the active sites of proteins or peptides to bind to or decompose pollutants. However, existing protein-based active substances (such as enzymes) face stability issues in practical applications. They are highly sensitive to environmental conditions (such as pH and temperature) and are susceptible to denaturation, aggregation, or browning during storage and use due to oxidative factors, leading to reduced activity and limiting their effectiveness.

[0005] Therefore, how to develop an air purification agent with excellent stability, capable of broadly removing a variety of chemical and biological pollutants (especially small molecule polar substances such as formaldehyde and amines), and safe and without secondary pollution is a technical problem that needs to be solved in this field. Summary of the Invention

[0006] To address the shortcomings of existing technologies, this invention provides a small molecule protein-based environmental air disinfection and deodorization preparation and its application, which solves the problems of poor stability, low removal efficiency of small molecule polar substances such as formaldehyde and amines, and easy generation of secondary pollution in existing air purification preparations.

[0007] To address the above problems, the present invention provides the following technical solution:

[0008] This invention provides a small molecule protein-based environmental air disinfection and deodorization preparation, employing the following technical solution:

[0009] A small molecule protein-based environmental air disinfection and deodorization agent, made from raw materials comprising the following parts by weight:

[0010] Core protein components: 50.0-80.0 parts;

[0011] Sodium citrate 1.0-5.0 parts;

[0012] Sorbitol 2.0-15.0 parts;

[0013] Anti-browning agent 0.05-0.3 parts;

[0014] L-Sodium ascorbate 5.0-15.0 parts;

[0015] Riboflavin 0.01-0.2 parts;

[0016] 5.0-20.0 parts of cyclodextrin or cyclodextrin derivatives;

[0017] Surfactant 0.05-0.5 parts;

[0018] Preservative: 0.1-0.8 parts;

[0019] The remainder is deionized water.

[0020] By employing the above technical solution, the components of this formulation synergistically construct a stable system. The core protein component is the main active substance that achieves the disinfection and deodorization functions. In the formulation, sodium citrate, sorbitol, and an anti-browning agent construct an anti-browning buffer environment in aqueous solution to inhibit the degradation of the core protein component. Simultaneously, cyclodextrin or cyclodextrin derivatives act as inclusion agents to encapsulate the core protein component and riboflavin. Sodium L-ascorbate acts as an antioxidant. The above components work together to control browning and inactivation of the formulation caused by inter-component interactions (such as oxidation and Maillard reactions) during storage and use.

[0021] Preferably, the anti-browning agent is selected from one or a combination of sodium sulfite or sodium metabisulfite;

[0022] The cyclodextrin or cyclodextrin derivative is selected from one or a combination of two of β-cyclodextrin or hydroxypropyl-β-cyclodextrin;

[0023] The surfactant is selected from one or a combination of two of polysorbate 80 or sucrose fatty acid esters;

[0024] The preservative is selected from one or a combination of potassium sorbate or a compound of phenoxyethanol and ethylhexylglycerin.

[0025] Preferably, the core protein component is an enzymatic hydrolysis product of soy protein isolate or whey protein isolate.

[0026] By adopting the above technical solution, the molecular weight of the core protein component is controlled within the range of small molecule peptides ≤20kDa, giving the active substance a larger specific surface area and more exposed active groups. This enhances its ability to capture and bind gaseous chemical pollutants (formaldehyde, ammonia, benzene, etc.) and strengthens its ability to destroy microbial cell structures.

[0027] Preferably, the formulation has the following technical features:

[0028] The pH value of the preparation is 6.0-8.0;

[0029] The sodium citrate, the sorbitol, and the anti-browning agent together constitute an anti-browning buffer environment in the formulation.

[0030] Preferably, the core protein component is prepared by a method comprising the following steps:

[0031] S01. Weigh the protein raw materials and add them to the deionized water to prepare a protein emulsion. Heat the emulsion at 85-95°C for 15-20 minutes to decompose the protein and obtain the decomposed protein emulsion.

[0032] S02. Cool the helical protein emulsion to 50-60℃, adjust the pH to 8.0-9.0, add alkaline protease and perform the first isothermal stirring hydrolysis to obtain the first enzymatic hydrolysate;

[0033] S03. Adjust the pH of the first enzymatic hydrolysate to 6.5-7.5, adjust the temperature to 45-55℃, add flavor protease and perform a second isothermal stirring hydrolysis to obtain the second enzymatic hydrolysate.

[0034] S04. The secondary enzymatic hydrolysate is rapidly heated to 95-100°C and held for 10-15 minutes to inactivate the enzyme, thereby obtaining an inactivated enzymatic hydrolysate.

[0035] S05. After cooling the inactivated enzyme hydrolysate to room temperature, centrifuge to collect the supernatant, separate it using an ultrafiltration membrane system with a molecular weight cutoff of 20 kDa, collect the filtrate that has permeated through the membrane and dry it.

[0036] By employing the above technical solution, this preparation method uses a two-stage tandem enzymatic hydrolysis process involving high-temperature unwinding and alkaline, flavor-enhancing proteases to degrade large-molecule proteins into small-molecule peptides. Step S05 uses an ultrafiltration membrane system with a molecular weight cutoff of 20 kDa for separation, limiting the molecular weight distribution of the final product (i.e., the core protein component) to ≤20 kDa.

[0037] Preferably, the parameters of the preparation method satisfy:

[0038] In step S01, the mass-volume concentration of the helical protein emulsion is 10%-15% (w / v);

[0039] In step S02, the amount of alkaline protease added is 1500-3000 U / g substrate protein, and the hydrolysis time is 60-120 minutes.

[0040] In step S03, the amount of flavor protease added is 1000-2500 U / g substrate protein, and the hydrolysis time is 60-120 minutes.

[0041] Preferably, the formulation is prepared by the following steps:

[0042] S1. Use deionized water, which accounts for approximately 80% of the total volume, as the base solution;

[0043] S2. Add sodium citrate, sorbitol and anti-browning agent to the base liquid in sequence, stir until the solid is completely dissolved, establish an anti-browning buffer environment and obtain a buffer solution;

[0044] S3. Add the L-ascorbic acid sodium, the riboflavin, and the cyclodextrin or cyclodextrin derivative to the buffer solution in sequence, and stir continuously until completely dissolved to obtain a functional solution;

[0045] S4. Slowly add the core protein component to the functional solution and stir to fully disperse and dissolve the core protein component until the solution is homogeneous and transparent to obtain a protein mixture.

[0046] S5. Add the surfactant and the preservative to the protein mixture in sequence, stir until homogeneous, and obtain the solution to be adjusted.

[0047] S6. Detect and adjust the pH value of the solution to be adjusted, and finally use deionized water to make up to the total volume of the preparation to obtain the formulation.

[0048] Preferably, in step S2, the order of adding each component is as follows: first add the sodium citrate and the sorbitol, dissolve them, and then add the anti-browning agent;

[0049] In step S4, the cyclodextrin or cyclodextrin derivative has been completely dissolved before the core protein component is added.

[0050] By adopting the above technical solution, the formulation preparation process employs a strategy of controlling the order of ingredient addition. The sequence of step S2 ensures the pre-establishment of an anti-browning buffer environment; the limitation of step S4 ensures the pre-dissolution of the inclusion agent. Adding the core protein component to the established buffer environment and inclusion system is a key step in stabilizing the protein component during dissolution.

[0051] This invention also provides the application of a small molecule protein-based environmental air disinfection and deodorization agent in removing indoor air chemical pollutants, as well as its application in air disinfection and virus inactivation, employing the following technical solutions:

[0052] The formulation is used to remove indoor air chemical pollutants, including formaldehyde, ammonia, benzene, and total volatile organic compounds.

[0053] The preparation is used for air disinfection and virus inactivation, to kill Staphylococcus aureus, Escherichia coli, Candida albicans, or to inactivate influenza A virus and adenovirus.

[0054] By employing the above technical solution, the formulation, with its stable ≤20kDa small molecule core protein component, achieves pollutant removal and biological disinfection functions. Its exposed active groups (such as amino groups) chemically bind or react with formaldehyde, ammonia, etc., achieving the removal of chemical pollutants. The peptide structure can act on bacterial cell structures or viral activity, achieving biological disinfection and inactivation.

[0055] This invention provides a small molecule protein-based environmental air disinfection and deodorization formulation and its application. It has the following beneficial effects:

[0056] 1. This invention provides a core protein component with a molecular weight of less than or equal to 20 kDa, which enables the active substance to have a larger specific surface area and more exposed active groups, thereby enhancing its ability to capture and bind to indoor air chemical pollutants and achieving the removal of pollutants. At the same time, its peptide structure can act on bacterial cell structure or viral activity, thereby achieving the disinfection and inactivation function of microorganisms and viruses, and endowing the preparation with multifunctional air purification function.

[0057] 2. This invention constructs an anti-browning buffer environment in the formulation through the synergistic effect of sodium citrate, sorbitol, and an anti-browning agent, and stabilizes the core protein component and riboflavin by using cyclodextrin or cyclodextrin derivatives as inclusion agents. This dual mechanism of buffer environment and inclusion controls the degradation, browning, or aggregation of components caused by oxidation or Maillard reaction during storage and use, thereby maintaining the bioactivity of the core protein component and a uniform and transparent solution state, ensuring the stability of the formulation.

[0058] 3. This invention constructs a stable system using sodium citrate, sorbitol, anti-browning agent, and L-ascorbic acid sodium excipients, and uses core protein components for pollutant removal and biological disinfection. The degradation products of this preparation are water, carbon dioxide, and harmless salts. The preparation itself does not introduce new volatile chemicals or harmful residues, thus avoiding secondary pollution. Detailed Implementation

[0059] The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the specification of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0060] Preparation Examples 1-3:

[0061] Preparation Example 1:

[0062] This preparation example provides a method for preparing a core protein component, including the following steps:

[0063] Weigh out soy protein isolate, add deionized water, and prepare a protein emulsion with a mass-volume concentration of 10% (w / v).

[0064] The protein emulsion was heated in a water bath at 85°C for 15 minutes to decompose the protein.

[0065] Cool the feed solution to 50°C, adjust the pH of the feed solution to 8.0 using 2M NaOH solution, add 1500 U / g of alkaline protease based on the weight of the substrate protein, and hydrolyze at a constant temperature with stirring for 60 minutes.

[0066] Adjust the pH of the feed solution to 6.5 using 1M HCl solution, adjust the temperature to 45℃, add 1000U / g of flavor protease based on the weight of the substrate protein, and hydrolyze at a constant temperature with stirring for 60 minutes.

[0067] The enzyme hydrolysate was rapidly heated to 95°C and held for 10 minutes to inactivate the enzyme.

[0068] After cooling the inactivated enzyme hydrolysate to room temperature, centrifuge it, take the supernatant, and separate it using an ultrafiltration membrane system with a molecular weight cutoff of 20 kDa. Collect the filtrate that permeates through the membrane.

[0069] The collected filtrate was freeze-dried to obtain the core protein component peptide powder, which was then set aside for later use.

[0070] Preparation Example 2:

[0071] This preparation example provides a method for preparing a core protein component, including the following steps:

[0072] Weigh out soy protein isolate, add deionized water, and prepare a protein emulsion with a mass-volume concentration of 12.5% ​​(w / v).

[0073] The protein emulsion was heated in a water bath at 90°C for 15 minutes to decompose the protein.

[0074] Cool the solution to 55°C, adjust the pH of the solution to 8.5 using 2M NaOH solution, add 2000 U / g of alkaline protease based on the weight of the substrate protein, and hydrolyze at a constant temperature with stirring for 90 minutes.

[0075] Adjust the pH of the feed solution to 7.0 using 1M HCl solution, adjust the temperature to 50℃, add 1500U / g of flavor protease based on the weight of the substrate protein, and hydrolyze at a constant temperature with stirring for 90 minutes.

[0076] The enzyme hydrolysate was rapidly heated to 95°C and held for 15 minutes to deactivate the enzyme.

[0077] After cooling the inactivated enzyme hydrolysate to room temperature, centrifuge it, take the supernatant, and separate it using an ultrafiltration membrane system with a molecular weight cutoff of 20 kDa. Collect the filtrate that permeates through the membrane.

[0078] The collected filtrate was freeze-dried to obtain the core protein component peptide powder, which was then set aside for later use.

[0079] Preparation Example 3:

[0080] This preparation example provides a method for preparing a core protein component, including the following steps:

[0081] Weigh whey protein isolate (or soy protein isolate), add deionized water, and prepare a protein emulsion with a mass-volume concentration of 15% (w / v).

[0082] The protein emulsion was heated in a water bath at 95°C for 20 minutes to decompose the protein.

[0083] Cool the feed solution to 60℃, adjust the pH of the feed solution to 9.0 using 2M NaOH solution, add 3000 U / g of alkaline protease based on the weight of the substrate protein, and hydrolyze at a constant temperature with stirring for 120 minutes.

[0084] Adjust the pH of the feed solution to 7.5 using 1M HCl solution, adjust the temperature to 55℃, add 2500 U / g of flavor protease based on the weight of the substrate protein, and hydrolyze at a constant temperature with stirring for 120 minutes.

[0085] The enzyme hydrolysate was rapidly heated to 100°C and held for 15 minutes to inactivate the enzyme.

[0086] After cooling the inactivated enzyme hydrolysate to room temperature, centrifuge it, take the supernatant, and separate it using an ultrafiltration membrane system with a molecular weight cutoff of 20 kDa. Collect the filtrate that permeates through the membrane.

[0087] The collected filtrate was freeze-dried to obtain the core protein component peptide powder, which was then set aside for later use.

[0088] Example 1:

[0089] This embodiment provides a small molecule protein-based environmental air disinfection and deodorization preparation. The preparation method, by weight, includes the following steps:

[0090] S1. Take approximately 80% of the total volume of the target solution of deionized water as the base solution;

[0091] S2. Add 1.0 part sodium citrate, 2.0 part sorbitol, and 0.05 part sodium sulfite to the base solution in sequence, and stir until the solid is completely dissolved to establish an anti-browning buffer environment;

[0092] S3. Add 5.0 parts of L-ascorbic acid sodium, 0.01 parts of riboflavin, and 5.0 parts of β-cyclodextrin to the above solution in sequence, and continue stirring until completely dissolved;

[0093] S4. Slowly add 50.0 parts of the core protein component prepared in Example 1, and stir to fully disperse and dissolve it until the solution is homogeneous and transparent;

[0094] S5. Add 0.05 parts of polysorbate 80 (Tween 80) and 0.1 parts of potassium sorbate to the solution in sequence, and stir until well mixed;

[0095] S6. Use 1M HCl or 2M NaOH solution to detect and adjust the pH of the solution to 6.0. Finally, use deionized water to bring the volume to the target level to obtain the deodorizing agent.

[0096] Example 2:

[0097] This embodiment provides a small molecule protein-based environmental air disinfection and deodorization preparation. The preparation method, by weight, includes the following steps:

[0098] S1. Take approximately 80% of the total volume of the target solution of deionized water as the base solution;

[0099] S2. Add 3.0 parts sodium citrate, 8.0 parts sorbitol, and 0.15 parts sodium sulfite to the base solution in sequence, and stir until the solid is completely dissolved to establish an anti-browning buffer environment;

[0100] S3. Add 10.0 parts of L-ascorbic acid sodium, 0.1 parts of riboflavin, and 10.0 parts of hydroxypropyl-β-cyclodextrin to the above solution in sequence, and stir continuously until completely dissolved;

[0101] S4. Slowly add 65.0 parts of the core protein component prepared in Example 2, and stir to fully disperse and dissolve it until the solution is homogeneous and transparent;

[0102] S5. Add 0.2 parts of polysorbate 80 (Tween 80) and 0.5 parts of phenoxyethanol and ethylhexylglycerin compound to the solution in sequence, and stir until homogeneous;

[0103] S6. Use 1M HCl or 2M NaOH solution to detect and adjust the pH of the solution to 7.0. Finally, use deionized water to bring the volume to the target level to obtain the deodorizing agent.

[0104] Example 3:

[0105] This embodiment provides a small molecule protein-based environmental air disinfection and deodorization preparation. The preparation method, by weight, includes the following steps:

[0106] S1. Take approximately 80% of the total volume of the target solution of deionized water as the base solution;

[0107] S2. Add 5.0 parts sodium citrate, 15.0 parts sorbitol, and 0.3 parts sodium metabisulfite to the base solution in sequence, and stir until the solid is completely dissolved to establish an anti-browning buffer environment;

[0108] S3. Add 15.0 parts of L-ascorbic acid sodium, 0.2 parts of riboflavin, and 20.0 parts of β-cyclodextrin to the above solution in sequence, and stir continuously until completely dissolved;

[0109] S4. Slowly add 80.0 parts of the core protein component prepared in Example 3, and disperse and dissolve it fully while stirring until the solution is homogeneous and transparent;

[0110] S5. Add 0.5 parts of sucrose fatty acid ester and 0.8 parts of phenoxyethanol and ethylhexylglycerol complex to the solution in sequence, and stir until homogeneous;

[0111] S6. Use 1M HCl or 2M NaOH solution to detect and adjust the pH of the solution to 8.0. Finally, use deionized water to bring the volume to the target level to obtain the deodorizing agent.

[0112] Comparative Examples 1-5:

[0113] Comparative Example 1:

[0114] Compared with Example 2, the difference is that the core protein component added in step S4 is replaced with an equal weight of soybean peptides prepared by conventional hydrochloric acid hydrolysis process, while the remaining steps and raw materials are the same.

[0115] Comparative Example 2:

[0116] Compared with Example 2, the difference is that the core protein component added in step S4 is replaced with an equal weight of unhydrolyzed soy protein isolate powder, while the remaining steps and raw materials are the same.

[0117] Comparative Example 3:

[0118] The difference from Example 2 is that riboflavin is not added in step S3, while the other steps and raw materials are the same.

[0119] Comparative Example 4:

[0120] Compared with Example 2, the difference is that sodium sulfite is not added in step S2, while the other steps and raw materials are the same.

[0121] Comparative Example 5:

[0122] The difference from Example 2 is that polysorbate 80 (Tween 80) is not added in step S5, while the other steps and raw materials are the same.

[0123] Test Examples 1-4:

[0124] Test Example 1:

[0125] This test case aims to verify the broad-spectrum purification capability of the finished formulation for typical indoor air pollutants.

[0126] Test subject:

[0127] Samples prepared in Examples 1, 2, and 3, and Comparative Examples 1, 2, 3, 4, and 5.

[0128] Experimental instruments and conditions: 3.0m³ environmental test chamber; high-precision thermometer and hygrometer; pollutant concentration detector (PID and electrochemical sensor); pollutant standard gas source.

[0129] Experimental environment: temperature (25.0±1.0)℃, relative humidity (50±5)%RH.

[0130] Experimental steps:

[0131] The 3.0m³ environmental test chamber was cleaned until the background concentration of each pollutant in the chamber was lower than the instrument detection limit.

[0132] Start the cabin circulation fan and inject pollutants through a standard air source to bring the pollutants in the cabin to the initial concentration shown in Table 1. Stable for 30 minutes;

[0133] Using a standard spray device, atomize 10.0 mL of the sample to be tested and spray it into the test chamber;

[0134] After the sample has been exposed to air for 24 hours (t=24h), the detection port is opened to measure the residual concentration of each pollutant in the chamber. );

[0135] Another blank control chamber was set up without spraying any sample. Under the same conditions (t=24h), the concentration after natural decay was measured. ), used to correct calculations;

[0136] Removal rate ( Calculate using the following formula:

[0137] ;

[0138] Test data:

[0139] Table 1: Comprehensive Odor Removal Performance Test Data

[0140]

[0141] in conclusion:

[0142] Table 1 shows that the formulations in Examples 1, 2, and 3, within the set parameter range, all exhibited removal effects on four pollutants: formaldehyde, ammonia, benzene, and TVOC, with removal rates ranging from 80.9% to 94.2%, confirming that the formulations have broad-spectrum deodorizing capabilities.

[0143] By analyzing the data from the proportional group, the synergistic mechanism of this scheme can be deconstructed:

[0144] The formaldehyde removal rates of Comparative Examples 1 and 2 were 55.0% and 42.0%, respectively, and the ammonia removal rates were 61.0% and 52.0%, respectively, lower than those of Example 2 (94.2% and 91.2%). This result indicates that conventional macromolecular proteins or acid-hydrolyzed proteins do not possess the corresponding deodorizing ability. This method, through a two-step enzymatic hydrolysis and 20 kDa ultrafiltration process used in Examples 1 to 3, achieved the desired deodorization. Small molecule peptides with -COOH active sites are the basis for Schiff base reactions and acid-base neutralization.

[0145] The data from Example 2 differed from those of Comparative Example 3, with Comparative Example 3 showing a formaldehyde removal rate of 73.0%, lower than the 94.2% of Example 2. This indicates that formaldehyde removal relies on a dual pathway: approximately 73.0% of the removal rate is attributed to the Schiff base reaction between the amino groups on the core protein component and formaldehyde; the remaining 21.2% of the removal efficiency is attributed to the catalytic reduction pathway of L-ascorbic acid sodium reducing formaldehyde to methanol under the catalysis of riboflavin.

[0146] Compared to Example 2, Comparative Example 5 showed similar removal rates for water-soluble formaldehyde and ammonia. However, Comparative Example 5's removal rates for hydrophobic benzene and TVOC were 15.0% and 48.0%, respectively, lower than Example 2's 83.5% and 86.9%. This indicates that the mechanism of action of the mass transfer enhancer polysorbate-80 is as follows: as a surfactant, it reduces the surface tension of droplets, disrupts the gas-liquid interface barrier, and allows non-polar molecules such as benzene and TVOC to enter the aqueous phase, where they are then encapsulated by the cyclodextrin in the formulation.

[0147] Comparative Example 4 showed the same removal rate as Example 2 in this test, indicating that sodium sulfite does not participate in the deodorization reaction within 24 hours, and its function is not reflected in removal efficiency. This set of data provides a distinction for subsequent stability tests to verify its anti-browning function.

[0148] This technical solution achieves the removal of both polar and non-polar pollutants through the synergistic effect of multiple components, including small molecule peptide active sites, catalytic reduction systems, supramolecular inclusion, and enhanced mass transfer.

[0149] Test Example 2:

[0150] This test case aims to verify the chemical appearance stability and biological stability of the finished formulation under accelerated storage conditions.

[0151] Test subjects: Example 2, Comparative Example 4, Comparative Example 6.

[0152] Experimental steps:

[0153] Chemical stability test:

[0154] Take sample solutions from Example 2 and Comparative Example 4 respectively, and place them in transparent sample bottles and seal them.

[0155] Place the sample vials in a constant temperature incubator at 45℃ (±1.0℃) and store them in the dark for 14 days.

[0156] Samples were taken on day 0 and day 14, and the absorbance of the samples at a wavelength of 420 nm (A420) was measured using a spectrophotometer. This absorbance value was used to characterize the degree of browning of the solution.

[0157] Biological stability testing:

[0158] A mixed bacterial suspension was used as the challenge strain, containing Staphylococcus aureus (ATCC6538), Escherichia coli (ATCC8739), Pseudomonas aeruginosa (ATCC9027), Aspergillus niger (ATCC16404), and Candida albicans (ATCC10231), and the concentration of the mixed bacterial suspension was adjusted to approximately 1.0 x 10⁻⁶. 8 CFU / mL;

[0159] Samples from Example 2 and Comparative Example 6 were inoculated with a mixed bacterial suspension at a ratio of 100:1 (v / v) to ensure that the initial total bacterial count in the samples was not less than 1.0 x 10⁻⁶. 5 CFU / mL;

[0160] The inoculated samples were incubated at (25±1)℃.

[0161] Samples were taken on days 7 and 14 after inoculation, and the total number of surviving colonies (CFU / mL) in the samples was determined by plate counting.

[0162] Test data:

[0163] Table 2: Chemical Stability Test Data

[0164]

[0165] Table 3: Biostability Test Data

[0166]

[0167] in conclusion:

[0168] The data in Tables 2 and 3 confirm that the formulation of this technical solution possesses both chemical and biological stability.

[0169] Chemical stability analysis: Table 2 shows that after accelerated storage at 45°C for 14 days, Comparative Example 4's A420 reading increased from 0.053 to 0.491, and its appearance turned brown. In contrast, Example 2's A420 reading only slightly increased from 0.051 to 0.068, remaining clear and slightly yellow. This phenomenon is due to the fact that the formulation system is rich in the core protein component (small molecule peptide), and its terminal and side chain free amino groups (… Sodium sulfite is a highly reactive group that readily undergoes a Maillard reaction with trace amounts of carbonyl compounds present in the system, leading to browning. In Comparative Example 4, the absence of sodium sulfite meant this reaction was not inhibited. In Example 2, sodium sulfite, acting as an anti-browning agent, blocked the initiation step of the Maillard reaction through addition to the carbonyl group, ensuring the chemical appearance stability of the formulation during storage.

[0170] Biostability analysis: Table 3 shows that in Comparative Example 6, the total bacterial count continued to increase after inoculation with the mixed bacterial solution, indicating that the formulation itself does not possess antibacterial ability and is prone to microbial contamination. In Example 2, after inoculation with the same concentration of bacterial solution, the total bacterial count decreased to <10 CFU / mL at both 7 and 14 days, meeting the standards for the preservative challenge test. This indicates that the phenoxyethanol and ethylhexylglycerin complex, as a biostabilizer, is necessary to maintain the biosafety of the formulation during storage and use.

[0171] Based on the data from Test Example 1, Comparative Example 4, while possessing initial deodorizing properties, lacked chemical storage stability. This technical solution addresses the browning and microbial growth issues of small molecule peptide preparations during storage by introducing sodium sulfite and a biological stabilizer.

[0172] Test Example 3: Comparison Test of Antibacterial and Antifungal Effects

[0173] This test case aims to verify the bactericidal effect of the formulation on representative bacteria and fungi, and to analyze the molecular weight of the core protein components and the influence of mass transfer enhancers on the bactericidal effect through a comparative group analysis.

[0174] Test subjects and bacterial strains:

[0175] Test subject: The finished products prepared according to the formulations of Example 2, Comparative Example 2, and Comparative Example 5 were prepared into a 5.0% (w / v) test solution using sterile deionized water.

[0176] Tested bacterial strains: Staphylococcus aureus (ATCC6538), Escherichia coli (ATCC8739), Candida albicans (ATCC10231).

[0177] Experimental steps:

[0178] Preparation of bacterial suspension: Staphylococcus aureus, Escherichia coli, and Candida albicans were inoculated onto nutrient agar or Sabouraud agar medium, respectively, and eluted with physiological saline to prepare a suspension with a concentration of approximately 1.0 x 10⁻⁶. 8 CFU / mL bacterial suspension;

[0179] Bactericidal test: A quantitative bactericidal test of the suspension was conducted according to the "Disinfection Technical Specifications". Take 4.0 mL of 5.0% test solution, add 1.0 mL of bacterial suspension, and mix well;

[0180] Contact time: The mixture of sample and bacterial suspension was incubated at (25±1)℃ for 5.0 minutes;

[0181] Neutralization and counting: After the reaction time is reached, immediately aspirate 1.0 mL of the mixture into 9.0 mL of sterile neutralizing agent (containing lecithin and Tween), and shake to neutralize for 10 minutes;

[0182] Take the neutralized diluted solution and count the viable bacteria (CFU / mL) using the plate pour method.

[0183] A separate negative control group was set up using physiological saline instead of the test solution, and the initial colony count was determined.

[0184] Kill rate ( Calculate using the following formula:

[0185] ;

[0186] Test data:

[0187] Table 4: Comparison Test Data of Antibacterial and Antifungal Effects

[0188]

[0189] in conclusion:

[0190] Table 4 shows that Example 2 achieved a kill rate of more than 99.9% against Staphylococcus aureus, Escherichia coli, and Candida albicans after 5 minutes of action, demonstrating broad-spectrum bactericidal ability.

[0191] Comparative Example 2 used unhydrolyzed macromolecular proteins, which achieved bactericidal rates of 51.05% to 64.88% against the three bacterial strains. A comparison of the data from Example 2 and Comparative Example 2 indicates that the bactericidal activity of the formulation does not originate from the macromolecular protein. The small molecule peptide components (≤20 kDa) obtained through enzymatic hydrolysis and ultrafiltration in this formulation possess biological activities not found in the original protein. This activity is achieved through mechanisms similar to antimicrobial peptides (such as cell membrane penetration or metabolic interference).

[0192] Comparative Example 5, lacking polysorbate 80, exhibited a lower bactericidal rate (74.83%-87.91%) than Example 2. This indicates that polysorbate 80 played a synergistic role in the system. As a surfactant, the assisting small molecule peptide disrupted the cell membrane or cell wall structure of microorganisms, enhancing the bactericidal efficiency of the formulation. This function represents another synergistic mechanism beyond the mass transfer enhancement effect observed in Test Example 1.

[0193] Test Example 4: Comparison Test of Virus Inactivation Effect

[0194] This test case aims to verify the inactivation effect of the formulation on enveloped viruses (H1N1 influenza A virus) and non-enveloped viruses (human adenovirus type 5), and to analyze the role of mass transfer enhancers (surfactants) and small molecule peptide components in the inactivation process through a comparative group.

[0195] Test subjects, viruses, and cells:

[0196] Test subject: The finished products prepared according to the formulations of Example 2, Comparative Example 2, and Comparative Example 5 were prepared into a 5.0% (w / v) test solution using sterile deionized water.

[0197] Tested viruses: H1N1 influenza A virus (ATCCVR-1469); human adenovirus type 5 (ATCCVR-5).

[0198] Host cells: MDCK cells (for H1N1); HEK293 cells (for Adeno-5).

[0199] Experimental steps:

[0200] Virus suspension preparation: Prepare virus stock solution using the corresponding host cells and determine the titer;

[0201] Inactivation test: According to the "Disinfection Technical Specifications", a quantitative virus suspension killing test was conducted. At (25±1)℃, 0.5 mL of the test solution was mixed with 0.5 mL of virus suspension containing organic matter (5% fetal bovine serum).

[0202] Action time: The mixture is allowed to act for 30.0 minutes;

[0203] Neutralization and titration: Once the reaction time has elapsed, immediately dilute the mixture 10-fold with pre-cooled cell maintenance medium (MEM) (to terminate the reaction) and seed it into 96-well cell culture plates;

[0204] Culture and observation: Incubate at 37℃ and 5% CO2 for 5-7 days and observe the cytopathic effect (CPE).

[0205] Calculation: The viral titer (lgTCID50) was calculated using the Karber method, and the logarithmic value of viral inactivation was also calculated.

[0206] Inactivation logarithm = Virus control group titer (lgTCID50) - Experimental group titer (lgTCID50).

[0207] Test data:

[0208] Table 5: Comparison Test Data on Virus Inactivation Efficacy

[0209]

[0210] in conclusion:

[0211] Table 5 shows that Example 2 has an inactivation log value greater than 4.00 against H1N1 influenza A virus and an inactivation log value of 3.13 against human adenovirus type 5, indicating that it has the ability to inactivate both enveloped and non-enveloped viruses.

[0212] Analysis of the inactivation mechanism of enveloped viruses: Comparative Example 2 and Comparative Example 5. Comparative Example 5 lacks polysorbate 80 (Tween 80), and its inactivation log value against H1N1 virus is only 1.25, lower than that of Example 2 (>4.00). H1N1 is an enveloped virus, and its envelope is a lipid bilayer. This difference in data indicates that polysorbate 80, as a surfactant, is a key component in the inactivation of this type of virus, and its mechanism lies in dissolving the lipid envelope of the virus, leading to viral lysis and inactivation.

[0213] Analysis of the inactivation mechanism of non-enveloped viruses: Comparative Example 2 and Comparative Example 2. Comparative Example 2 used a large molecular weight protein, whose inactivation log value against human adenovirus type 5 (non-enveloped, composed of a protein capsid) was only 0.85, lower than 3.13 in Example 2. This data indicates that the large molecular weight protein does not have the ability to inactivate the viral capsid. The small molecular weight peptide (≤20kDa) obtained in this preparation method can interfere with or destroy the protein capsid structure of the virus, thereby achieving inactivation of non-enveloped viruses.

[0214] This technical solution utilizes the synergistic effect of small molecule peptide components and surfactants to target the protein capsid of non-enveloped viruses and the lipid envelope of enveloped viruses, respectively, thereby achieving broad-spectrum inactivation of different types of viruses.

Claims

1. A small molecule protein-based environmental air disinfection and deodorization preparation, characterized in that, The formulation is made from raw materials comprising the following parts by weight: Core protein components: 50.0-80.0 parts; Sodium citrate 1.0-5.0 parts; Sorbitol 2.0-15.0 parts; Anti-browning agent 0.05-0.3 parts; L-Sodium ascorbate 5.0-15.0 parts; Riboflavin 0.01-0.2 parts; 5.0-20.0 parts of cyclodextrin or cyclodextrin derivatives; Surfactant 0.05-0.5 parts; Preservative: 0.1-0.8 parts; The remainder is deionized water; The core protein component is an enzymatic hydrolysis product of soy protein isolate or whey protein isolate. The core protein component is prepared by a method including the following steps: S01. Weigh the protein raw materials and add them to the deionized water to prepare a protein emulsion. Heat the emulsion at 85-95°C for 15-20 minutes to decompose the protein and obtain the decomposed protein emulsion. S02. Cool the helical protein emulsion to 50-60℃, adjust the pH to 8.0-9.0, add alkaline protease and perform the first isothermal stirring hydrolysis to obtain the first enzymatic hydrolysate; S03. Adjust the pH of the first enzymatic hydrolysate to 6.5-7.5, adjust the temperature to 45-55℃, add flavor protease and perform a second isothermal stirring hydrolysis to obtain the second enzymatic hydrolysate. S04. The secondary enzymatic hydrolysate is rapidly heated to 95-100°C and held for 10-15 minutes to inactivate the enzyme, thereby obtaining an inactivated enzymatic hydrolysate. S05. After cooling the inactivated enzyme hydrolysate to room temperature, centrifuge to collect the supernatant. Separate the supernatant using an ultrafiltration membrane system with a molecular weight cutoff of 20 kDa. Collect the filtrate that has permeated through the membrane and dry it to obtain the core protein component.

2. The small molecule protein-based environmental air disinfection and deodorization preparation according to claim 1, characterized in that: The anti-browning agent is selected from one or a combination of two of sodium sulfite or sodium metabisulfite; The cyclodextrin or cyclodextrin derivative is selected from one or a combination of two of β-cyclodextrin or hydroxypropyl-β-cyclodextrin; The surfactant is selected from one or a combination of two of polysorbate 80 or sucrose fatty acid esters; The preservative is selected from one or a combination of potassium sorbate or a compound of phenoxyethanol and ethylhexylglycerin.

3. The small molecule protein-based environmental air disinfection and deodorization preparation according to claim 1, characterized in that, The formulation is prepared by the following steps: S1. Use deionized water, which accounts for 80% of the total volume, as the base solution; S2. Add sodium citrate, sorbitol and anti-browning agent to the base liquid in sequence, stir until the solid is completely dissolved, establish an anti-browning buffer environment and obtain a buffer solution; S3. Add the L-ascorbic acid sodium, the riboflavin, and the cyclodextrin or cyclodextrin derivative to the buffer solution in sequence, and stir continuously until completely dissolved to obtain a functional solution; S4. Slowly add the core protein component to the functional solution and stir to fully disperse and dissolve the core protein component until the solution is homogeneous and transparent to obtain a protein mixture. S5. Add the surfactant and the preservative to the protein mixture in sequence, stir until homogeneous, and obtain the solution to be adjusted. S6. Detect and adjust the pH value of the solution to be adjusted, and finally use deionized water to make up to the total volume of the preparation to obtain the formulation.

4. The small molecule protein-based environmental air disinfection and deodorization preparation according to claim 3, characterized in that: In step S2, the order of adding each component is as follows: first add the sodium citrate and the sorbitol, dissolve them, and then add the anti-browning agent; In step S4, the cyclodextrin or cyclodextrin derivative has been completely dissolved before the core protein component is added.

5. The small molecule protein-based environmental air disinfection and deodorization preparation according to claim 1, characterized in that, The pH value of the formulation is 6.0-8.0; the sodium citrate, the sorbitol, and the anti-browning agent together constitute an anti-browning buffer environment in the formulation.

6. The small molecule protein-based environmental air disinfection and deodorization preparation according to claim 1, characterized in that, The parameters of the preparation method satisfy: In step S01, the mass-volume concentration of the helical protein emulsion is 10%-15% (w / v); In step S02, the amount of alkaline protease added is 1500-3000 U / g substrate protein, and the hydrolysis time is 60-120 minutes. In step S03, the amount of flavor protease added is 1000-2500 U / g substrate protein, and the hydrolysis time is 60-120 minutes.

7. The application of a small molecule protein-based environmental air disinfection and deodorizing agent according to any one of claims 1-6 in removing indoor air chemical pollutants, characterized in that, The chemical pollutants include formaldehyde, ammonia, benzene, and total volatile organic compounds.

8. The application of a small molecule protein-based environmental air disinfection and deodorization agent according to any one of claims 1-6 in air disinfection and virus inactivation, characterized in that, The formulation is used to kill Staphylococcus aureus, Escherichia coli, and Candida albicans, or to inactivate influenza A virus and adenovirus.

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