Antibacterial disinfection method using seaw and thymol in cooperation and product thereof

Abstract

The antibacterial disinfection product adopting SAEW cooperates with thymol, including independently assembled and step-by-step used quorum sensing inhibitor A and disinfectant B; the quorum sensing inhibitor A includes 5-methyl-2-isopropyl phenol and deionized water completely dissolved in an organic solvent, and the concentration of 5-methyl-2-isopropyl phenol in the quorum sensing inhibitor A is not less than 256 μg / mL; the disinfectant B includes micro-acidic electrolytic water with an effective chlorine concentration of 30 ppm; the quorum sensing inhibitor A is filled into a spray bottle to obtain spray A; the disinfectant B is filled into a spray bottle to obtain spray B. Through the combined application of SAEW and the quorum sensing inhibitor thymol from plants, a new type of composite disinfectant with high efficient synergy is obtained. The combination can significantly reduce the risk of traditional chemical disinfectants in bacterial drug resistance and pathogenicity improvement, show good antibacterial and antibiofilm effects, and can reduce the frequency of bacterial drug resistance mutation, and is expected to become an important means of clinical and public environmental anti-infection prevention strategy.

Description

Technical Field

[0001] This invention patent relates to the field of antibacterial disinfection technology of slightly acidic electrolyzed water, and in particular to the antibacterial disinfection method and products using SAEW in combination with thymol. Background Technology

[0002] Chemical disinfectants can be classified into nine categories based on their active ingredients: peroxides, aldehydes, alcohols, chlorine-containing disinfectants, iodine-containing disinfectants, phenols, ethylene oxide, biguanides, and quaternary ammonium salts. Given the potential environmental hazards posed by traditional disinfectant residues, slightly acidic electrolyzed water (SAEW) has been proposed as a new, safe, and environmentally friendly disinfectant. SAEW, also known as hypochlorous acid water, is a solution produced by electrolyzing dilute electrolytes (usually containing NaCl and / or HCl) in an electrolysis chamber without a diaphragm. SAEW solutions have a near-neutral pH (5.0-6.5), and the available chlorine exists almost entirely in the form of hypochlorous acid molecules, enabling it to kill various bacteria, fungi, and common viruses in a short time. Compared to other commonly used disinfectants such as sodium hypochlorite and 75% ethanol, SAEW does not leave disinfectant residues, does not corrode instrument surfaces, and is non-irritating to eyes, skin, and respiratory tract. Due to its broad-spectrum bactericidal properties, rapid and efficient disinfection, low cost, and environmental safety, SAEW has broad application prospects in cleaning and disinfection in the food, medical, environmental, and surface industries.

[0003] However, the potential harm caused by the irrational use of chemical disinfectants has attracted attention. Existing research indicates that the use of chemical disinfectants can induce the emergence of disinfectant-resistant bacteria, some of which exhibit cross-resistance to clinically available antimicrobial agents. Furthermore, chemical disinfectants can promote the transfer of bacterial resistance genes at environmental concentrations, exacerbating the resistance situation. The emergence and spread of these resistant strains seriously endanger public health and limit the selection of clinically available antimicrobial agents.

[0004] Furthermore, disinfectants can enhance the virulence of pathogenic microorganisms. For example, peracetic acid causes high levels of sublethal damage to the foodborne pathogen Listeria monocytogenes, and bacteria in this state exhibit upregulated expression of virulence genes and enhanced invasiveness to host cells. Chlorhexidine-induced resistant Gram-negative bacteria show upregulation of the virulence factors LPS assembly protein and fimbrial forming protein. Studies have shown that reactive oxygen species (ROS) generated during disinfectant damage may be an important means for pathogens to regulate virulence factors; that is, when ROS decreases to nonlethal levels, it is more likely to induce drug-resistant mutations in pathogens. Therefore, antiviral therapy has become a novel treatment strategy for combating multidrug-resistant bacterial infections in clinical practice.

[0005] Slightly acidic electrolyzed water (SAEW), like other chlorine-containing disinfectants, exerts its antibacterial effect through the oxidation potential of hypochlorous acid. However, its potential to promote bacterial resistance and virulence limits its application. Furthermore, the emergence of chlorine-resistant bacterial strains further restricts the antibacterial efficacy of SAEW. Summary of the Invention

[0006] The purpose of this invention is to solve the existing technical problems and provide an antibacterial disinfection method and product that uses slightly acidic electrolyzed water (SAEW) in combination with the plant extract thymol (5-methyl-2-isopropylphenol) to exhibit good antibacterial and anti-biofilm effects and can reduce the frequency of bacterial drug resistance mutations.

[0007] Antimicrobial and disinfectant products using SAEW in combination with thymol include separately assembled, step-by-step quorum sensing inhibitor A and disinfectant B;

[0008] Quorum sensing inhibitor A comprises 5-methyl-2-isopropylphenol and deionized water completely dissolved in an organic solvent, and the concentration of 5-methyl-2-isopropylphenol in quorum sensing inhibitor A is not less than 256 μg / mL;

[0009] Disinfectant B consists of slightly acidic electrolyzed water with an effective chlorine concentration of 30 ppm;

[0010] Group sensing inhibitor A is loaded into a spray bottle to obtain spray A;

[0011] Disinfectant B is filled into a spray bottle to obtain spray B.

[0012] Furthermore, the spray bottle can be a double-headed spray bottle, with one nozzle on one side for releasing spray A and the other nozzle on the other side for releasing spray B.

[0013] Preferably, the organic solvent in quorum sensing inhibitor A is ethanol with a mass fraction of 2-5%.

[0014] Preferably, the concentration of 5-methyl-2-isopropylphenol in quorum sensing inhibitor A is 256-1024 μg / mL.

[0015] Furthermore, the concentration of 5-methyl-2-isopropylphenol in quorum sensing inhibitor A is preferably 512 μg / mL.

[0016] Furthermore, the quorum sensing inhibitor A also contains 0.01–0.08% by mass of edible flavoring and 0.01–0.08% by mass of edible coloring.

[0017] Preferably, disinfectant B is prepared freshly using a slightly acidic electrolyzed water generator. Disinfectant B is prepared by electrolyzing NaCl and / or HCl solutions using a slightly acidic electrolyzed water generator.

[0018] The antibacterial disinfection method of the product described in this invention involves uniformly spraying spray A onto the area to be disinfected, allowing it to act for 10 minutes, and then uniformly spraying spray B onto the same area to be disinfected, thus completing the disinfection operation of the environment or medical device.

[0019] The antibacterial disinfection method of the product described in this invention uses quorum sensing inhibitor A as a mouthwash, which is left in the mouth for 10-30 seconds and then spat out. After rinsing with water, spray B is then applied to the mouth to clean the oral cavity, thus completing the oral disinfection operation for non-therapeutic purposes.

[0020] The antibacterial disinfection method of the product described in this invention involves using spray A to clean the oral cavity, applying it for 30-60 seconds, rinsing with water, and then using spray B to clean the oral cavity, thus completing an oral disinfection operation for non-therapeutic purposes.

[0021] Furthermore, for children, spray A can be evenly sprayed onto the surface of a cotton swab, and disinfection can be achieved by swirling the cotton swab in the mouth. After 30-60 seconds, rinse the mouth with water, then spray B evenly onto the surface of another cotton swab and repeat the swirling operation to complete the oral disinfection.

[0022] The preparation method of the product of the present invention includes:

[0023] 1) Preparation of spray A: Add solid thymol (5-methyl-2-isopropylphenol) to ethanol and stir until completely dissolved. Then add an aqueous solution of food flavoring and food coloring and stir well. Finally, add the remaining deionized water and stir well to obtain quorum sensing inhibitor A. Add quorum sensing inhibitor A to a spray bottle to obtain spray A.

[0024] 2) Preparation of spray B: Prepare a NaCl solution at a ratio of 1.875 g / L, electrolyze the NaCl solution using a slightly acidic water electrolysis generator to obtain 50 ppm SAEW, then dilute it with water to 30 ppm to obtain disinfectant B, and add disinfectant B to a spray bottle to obtain spray B.

[0025] The beneficial effects of this invention are: by combining SAEW with thymol, a quorum sensing inhibitor derived from plants, a novel composite disinfectant with highly efficient synergistic effects is obtained. This combination can significantly reduce the risks associated with the development of bacterial resistance and increased pathogenicity associated with traditional chemical disinfectants. Even low concentrations of SAEW exhibit good antibacterial and anti-biofilm effects, high safety, and can reduce the frequency of bacterial resistance mutations, making it a promising tool for infection control strategies in clinical settings and public environments. Attached Figure Description

[0026] Figure 1This is a graph showing the colony count results of the SAEW-assisted thymol removal (24-well plate) bacterial maturation biofilm experiment in Example 2 of the present invention;

[0027] Figure 2 This is a graph showing the colony count results of the SAEW synergistic thymol removal experiment for the mature bacterial biofilm of medical catheters in Example 2 of the present invention.

[0028] Figure 3 This is a graph showing the statistical results of ROS fluorescence intensity of the strain treated with SAEW and thymol in Example 3 of the present invention.

[0029] Figure 4 In the experiment of SAEW synergistically reducing bacterial virulence with thymol in Example 4 of this invention lasA, lasB, rhlA, pqsA and pqsE Statistical results of relative expression levels;

[0030] Figure 5 The graph shows the effect of 512 μg / mL thymol on the resistance mutation frequency of Pseudomonas aeruginosa PAO1 after high-concentration SAEW resistance treatment in Example 5 of this invention.

[0031] Figure 6 This is a photograph of the results of the acute skin toxicity test in mice in Example 6 of this invention;

[0032] Figure 7 These are photographs of the right eyes of mice in each group during the mouse eye stimulation experiment in Example 6 of this invention. Detailed Implementation

[0033] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments.

[0034] Example 1: Preparation of an antibacterial and disinfectant product using SAEW in synergy with thymol

[0035] 1) Preparation of spray A: 32, 64, 128, 256, 512 and 1024 μg / mL of 5-methyl-2-isopropylphenol (Shanghai Yuanye Biotechnology Co., Ltd.) solids were added to ethanol and stirred until completely dissolved. Then, aqueous solutions of food flavoring and food coloring were added and stirred evenly. Finally, the remaining water was added and stirred evenly to obtain quorum sensing inhibitor A for each group. Then, each group of quorum sensing inhibitor A was added to a spray bottle to obtain spray A.

[0036] The quorum sensing inhibitor A also includes: 2-5% ethanol, 0.01-0.08% food flavoring, 0.01-0.08% food coloring, and the balance being deionized water.

[0037] 2) Preparation of spray B: Prepare a NaCl solution at a ratio of 1.875 g / L, electrolyze the NaCl solution using a slightly acidic water electrolysis generator to obtain 50 ppm SAEW, then dilute it with pure water to 30 ppm to obtain disinfectant B, and add disinfectant B to a spray bottle to obtain spray B.

[0038] Disinfectant B consists of slightly acidic electrolyzed water with an effective chlorine concentration of 30 ppm.

[0039] Example 2: Thymol combined with slightly acidic electrolyzed water (SAEW) effectively removes mature bacterial biofilm.

[0040] 2.1 SAEW synergistically removes mature bacterial biofilms with thymol

[0041] Pseudomonas aeruginosa strain PAO1 (purchased from the National Center for Clinical Laboratories, National Health Commission, NCCL) was added to LB broth (OXOID, UK), and the McFarland turbidity was adjusted to 0.5 to obtain a bacterial suspension. 1 mL of this bacterial suspension was added to each well of a 24-well plate and incubated at 37°C for 12 hours in a CO2 incubator to form a mature biofilm. Before the experiment, the mature biofilm was rinsed with PBS (Guangzhou Kerong Biotechnology Co., Ltd.) to remove suspended bacteria.

[0042] In the experiment, the combined group first added 500 μL of quorum sensing inhibitor A containing 32, 64, 128, 256, 512, and 1024 μg / mL of thymol to each well, and allowed it to react for 10 minutes. After reaction, the solution was discarded, the cells were rinsed once with PBS, and then 500 μL of disinfectant B was added and treated for 10 minutes. After reaction, the solution was discarded, and 0.5% sodium thiosulfate solution (Shanghai Aladdin Biochemical Technology Co., Ltd.) was added to remove residual chlorine. Then, 1 mL of PBS was added to each well, and the cells were sonicated to release bacteria, obtaining a bacterial suspension. The bacterial suspension was then subjected to a 10-fold dilution of PBS. -1 -10 -7 After serial dilution, the cells were inoculated onto LB agar plates and incubated at 37°C for 16-18 hours before colony counting.

[0043] The difference between the single-drug group and the combination group is that only one quorum sensing inhibitor A or disinfectant B is added, and after 10 minutes of action, a 0.5% sodium thiosulfate solution is added to remove residual chlorine.

[0044] The control group differed from the combined group in that: 0.5% sodium thiosulfate solution was added directly to each well, followed by the addition of 1 mL of PBS to each well, and then the bacteria were sonicated to release them, obtaining a bacterial suspension. The bacterial suspension was then... -1 -10 -7 After serial dilution, the cells were inoculated onto LB agar plates and incubated at 37°C for 16-18 hours before colony counting.

[0045] like Figure 1 As shown, 32, 64, and 128 μg / mL (1 / 8xMIC - 1 / 8xMIC) thymol combined with 30 ppm SAEW did not show a combined effect; 256 μg / mL thymol combined with SAEW reduced PAO1 in the biofilm by 5.03 log CFU / mL, which was significantly better than the SAEW monotherapy group and the 256 μg / mL thymol monotherapy group, showing a statistically significant difference; 512 μg / mL thymol combined with SAEW reduced PAO1 in the biofilm by 5.73 log CFU / mL, which was close to the limit of detection (10). 2 Compared to the SAEW monotherapy group and the 512 μg / mL thymol monotherapy group, the effect was significantly improved, with statistically significant differences. Treatment of the biofilm with 1024 μg / mL thymol alone or with SAEW in synergy reduced the bacterial count in the biofilm to below the detection limit; therefore, the combined effect of thymol and SAEW could not be demonstrated in this example. Furthermore, due to the high concentration of thymol, it is easily precipitated, and the stability of population inhibitor A is poor.

[0046] The above results indicate that the application of thymol in combination with slightly acidic electrolyzed water (SAEW) can effectively remove mature bacterial biofilms, exhibiting good antibacterial and anti-biofilm effects. Furthermore, it was found that 256 μg / mL of thymol combined with SAEW can effectively remove biofilms, while 512 μg / mL of thymol combined with SAEW has the best effect. The bactericidal activity of 1024 μg / mL of thymol administered alone is below the detection limit.

[0047] 2.2 SAEW synergistically removes mature biofilm from medical catheters using thymol

[0048] According to the treatment method in 2.1, the 24-well plate was replaced with a 1 cm long medical catheter (Jiangsu Suyun Medical Equipment Co., Ltd.) for biofilm culture, and single-drug and combination experiments were conducted using population inhibitor A containing 128, 256, and 512 μg / mL thymol and disinfectant B containing 30 ppm, respectively.

[0049] The results are as follows Figure 2As shown, 128 μg / mL (1 / 2×MIC) thymol combined with 30 ppm SAEW did not show a combined effect; however, 256 μg / mL (1×MIC) and 512 μg / mL (2×MIC) thymol combined with 30 ppm SAEW showed better combined effects in clearing catheter biofilms. Specifically, 256 μg / mL thymol combined with SAEW could reduce the bacterial count in catheter biofilms by 2.7 log CFU / mL; and 512 μg / mL thymol combined with SAEW could reduce the bacterial count in catheter biofilms to below the detection limit.

[0050] Example 3: Analysis of Reactive Oxygen Species (ROS) Generation in Strains Treated with SAEW and Thymol

[0051] The reactive oxygen species (ROS) detection kit (Shanghai Beyotime Biotechnology Co., Ltd.) was used to detect the bacterial strain. PAO1 was cultured overnight in LB broth with shaking to obtain bacterial suspension. 8 mL of the bacterial suspension was centrifuged at 4000 rpm for 5 minutes, the supernatant was discarded, and the suspension was washed twice with PBS. Finally, the turbidity of the bacterial suspension was adjusted to OD600 of 0.3-0.4. Then, 12 μL of probe was mixed with 12 mL of bacterial suspension (1:1000 dilution of probe), and the mixture was transferred to a 15 mL centrifuge tube, wrapped in aluminum foil to protect from light, and incubated at 37°C for 20 minutes. The tube was inverted every 3-5 minutes to ensure that the probe and bacterial suspension were in full contact. After mixing the bacterial suspension and probe, the mixture was dispensed into 10 1.5 mL centrifuge tubes, 0.9 mL in each tube. After centrifugation at 4000 rpm for 5 minutes, the tubes were washed twice with PBS to remove free probe, and the experimental bacterial suspension loaded with probe was obtained.

[0052] Perform the following operations on the experimental bacterial culture:

[0053] The single-drug treatment groups were carried out according to the following protocols: 1) After centrifuging the bacterial cells at 4000 rpm for 5 minutes, add 1 mL of 30 ppm SAEW and treat the bacterial cells for 10 minutes; or 2) After centrifuging the bacterial cells at 4000 rpm for 5 minutes, add 1 mL of 128, 256, or 512 μg / mL of thymol and treat the bacterial cells for 10 minutes.

[0054] The combined group was treated as follows: First, 1 mL of 128, 256, and 512 μg / mL thymol was added to treat the bacterial cells for 10 minutes. After centrifugation at 4000 rpm for 5 minutes, the supernatant was discarded. Then, the bacterial cells were treated with 30 ppm SAEW for 10 minutes.

[0055] The positive control group used Rosup, the control reagent provided by the reactive oxygen species (ROS) detection kit, diluted 1:1000 by volume, and 1 mL was added to the mixed bacterial culture and treated for 10 minutes; the negative control group was treated with 1 mL of purified water for 10 minutes.

[0056] After processing, plate-laying and detection were performed, with 200 μL per well and 3 replicates per group. An excitation wavelength of 488 nm and an emission wavelength of 525 nm were used, with a gain of 100 / 80. The fluorescence intensity before and after stimulation was measured, and the data were recorded and analyzed. Figure 3 As shown, compared with the control group, the ROS level in the 30 ppm SAEW group was significantly increased; the ROS levels in the 128, 256, and 512 μg / mL thymol single-drug groups increased in a dose-dependent manner; compared with their respective single-drug groups, the ROS levels in each combination group were increased. Among them, the ROS level in the 256 μg / mL thymol and 30 ppm SAEW combination group was significantly higher than that in the 30 ppm SAEW group and the 256 μg / mL thymol group; when 512 μg / mL thymol was combined with SAEW, the data showed that the ROS increase was not as high as that in the single-drug group. This is because a better bactericidal effect was achieved in the combination, and the leakage and loss of intracellular fluorescent probes led to the ROS increase.

[0057] Example 4: Experiment on the reduction of bacterial virulence by SAEW in synergistic effect with thymol

[0058] RT-qPCR was used to investigate the changes in the expression levels of bacterial virulence genes under different drug combinations. This invention selected five known key virulence genes of PAO1 for analysis, including... lasA, lasB, rhlA, pqsA and pqsE The RT-qPCR experiment mainly includes three steps: RNA extraction, reverse transcription of RNA into cDNA, and cDNA qPCR. The research methods used in this invention are all classic RT-qPCR steps.

[0059] First, the treatments were performed according to the following groups: (1) Control group: no treatment was given; (2) Single-drug group: treated with 30 ppm SAEW and 512 μg / mL thymol respectively; (3) Combined group: treated with 512 μg / mL thymol first, and then treated with 30 ppm SAEW after a 10-minute interval. After treatment, the biofilm was broken to release the bacteria, which were then cultured in LB medium at 37°C to obtain a sufficient bacterial quantity (1×10⁻⁶). 8 CFU / mL was used for RNA extraction.

[0060] Using rpoB as the housekeeping gene, a 2 -ΔΔCtThe expression levels of virulence genes were calculated using a method that... RNA was extracted according to the RNA extraction kit (Baori Biotechnology (Beijing) Co., Ltd.), and cDNA was synthesized using the bacterial RNA Miniprep kit (Baori Biotechnology (Beijing) Co., Ltd.) and the PrimeScript™ II 1st Strand cDNA Synthesis Kit (Baori Biotechnology (Beijing) Co., Ltd.). TB Green... ® PremixEx Taq™ PCR amplification was performed using Tli-RNase HPlμs. The amplification primer sequences are shown in Table 1.

[0061] Table 1. Primer sequences

[0062]

[0063] Pseudomonas aeruginosa PAO1 can degrade elastin (a major component of connective tissue), which may be an important reason for its pathogenicity and proliferation in human tissues. LasA is an extracellular enzyme essential for its elastase activity. Extracellularly secreted LasB is considered an important virulence factor of Pseudomonas aeruginosa. It is also involved in the processing of LasA, thereby enhancing elastase activity. Figure 4 As shown, compared with the control group and the single-drug group, the combination group significantly reduced the virulence gene of Pseudomonas aeruginosa. lasA , lasB The relative expression levels, especially lasA ;right rhlA , pqsA , pqsE No significant effect. The above results indicate that the combined group reduced the effect caused by… lasA, lasB Induced bacterial virulence has promising applications.

[0064] Example 5: Thymol reduces the frequency of drug resistance mutations in Pseudomonas aeruginosa PAO1 against SAEW.

[0065] Two high concentrations of SAEW, 60 ppm and 80 ppm, were selected as experimental reagents to simulate the formation of drug-resistant bacteria induced by high concentrations of disinfectant.

[0066] Pick a single pure PAO1 colony and incubate it in 20 mL of MH broth (OXOID, UK) with shaking for 18-24 hours. Centrifuge at 3000 rpm for 10 minutes, resuspend the precipitate, and add it to 200 mL of MH broth. Incubate with shaking at 200 rpm for 6 hours, then centrifuge again at 3000 rpm for 10 minutes. Discard the supernatant and adjust the bacterial concentration to 1.0 × 10⁻⁶. 9CFU / mL, and at the same time prepare MH solid culture medium containing 512 μg / mL thymol for later use.

[0067] First, treat the bacteria with 60 ppm and 80 ppm SAEW disinfectant for 10 minutes. Then, spread the mixed suspension containing SAEW and bacteria onto blank MH solid medium and drug-containing MH solid medium, respectively. The reaction of SAEW was not terminated here to simulate the inhibitory effect of the two on the frequency of drug resistance mutation under the combined action. The spread plates were incubated at 37°C for 3 days.

[0068] Colony count results as follows Figure 5 As shown, the results indicate that 512 μg / mL thymol can significantly reduce the PAO1 resistance mutation rate induced by SAEW, and can reduce the resistance mutation frequency by 1×10⁻⁶. 3 The combined use of thymol and SAEW can reduce the frequency of bacterial drug resistance mutations and make it less likely to induce the formation of drug-resistant bacteria.

[0069] Example 6 Acute skin toxicity test and ocular irritation test in mice

[0070] The single-test limit method was used in accordance with the "Cosmetic Safety Technical Specifications" (2015 edition).

[0071] The application area was 10% of the mouse's body surface area, and the hair-removed area was 3cm × 3cm. The drugs were evenly applied to the shaved area on the mouse's back according to the following treatments: control group (1 mL deionized water), 1 mL of 512 μg / mL thymol, 1 mL of 30 ppm SAEW, and combination group (1 mL of 512 μg / mL thymol applied first, followed by 1 mL of 30 ppm SAEW). The mice were observed at 1 hour, 3 hours, 8 hours, and 24 hours after administration. The mice showed no significant changes in their general condition; their respiration, limb function, and behavior were all normal. Figure 6 As shown, in the acute skin toxicity test in mice, compared with the control group, no redness, damage or erythema was observed in the mouse skin within 1-24 hours after treatment in the single drug group and the combination group, proving that the combined application of SAEW and thymol is not skin irritating.

[0072] Mouse eye stimulation experiment: The experimental animals were 6 SPF grade mice in each group (half male and half female, females were non-pregnant and had never given birth), for a total of 4 groups. After 3 days of acclimatization, the mouse eye stimulation experiment was carried out.

[0073] During the experiment, the mouse eyeballs were gently opened, and the following solutions were instilled into the right eye of the mice: control group (10 μL deionized water), 10 μL 512 μg / mL thymol, 10 μL 30ppm SAEW, and a combination group (10 μL 512 μg / mL thymol was instilled first, followed by 10 μL 30ppm SAEW). The mouse eye was then passively closed for 1 second to prevent loss of the test substances. 10 μL of deionized water was instilled into the left eye of each group. The eyes were not rinsed within 24 hours after instillation. The mice's eyes were observed and photographed at 1 hour, 24 hours, and 48 hours after instillation. Observations showed that the condition of the left eyeballs in each group was similar to that of the right eyeball in the control group.

[0074] like Figure 7 As shown, in the mouse ocular irritation experiment, compared with the control group, no redness, swelling, inflammation, cloudiness, or tearing was observed in the eyes within 1-48 hours after treatment with 30 ppm SAEW, 512 μg / mL thymol, and the combined group. This proves that the combined application of SAEW and thymol is not ocular irritating.

[0075] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. An antibacterial disinfectant product employing SAEW in conjunction with thymol, characterized in that, The quorum sensing inhibitor A and the disinfectant B are independently assembled and used in steps. The quorum sensing inhibitor A comprises 5-methyl-2-isopropyl phenol and deionized water completely dissolved in an organic solvent, and the concentration of 5-methyl-2-isopropyl phenol in the quorum sensing inhibitor A is 256-512 ug / mL. The disinfectant B comprises micro-acidic electrolytic water with an effective chlorine concentration of 30 ppm. The quorum sensing inhibitor A is filled into a spray bottle to obtain a spray A. The disinfectant B is filled into a spray bottle to obtain a spray B. The disinfectant B is prepared by a micro-acidic electrolytic water generator.

2. The antiseptic disinfecting product of claim 1, wherein, The organic solvent in the quorum sensing inhibitor A is ethanol with a mass fraction of 2-5%.

3. The method of antibacterial disinfection of a product according to claim 1 or 2, characterized in that, The spray A is uniformly sprayed to the place to be disinfected, and after 10 minutes of action, the spray B is uniformly sprayed to the same place to be disinfected, thereby completing the disinfection operation of the environment or medical equipment.

4. The method of antibacterial disinfection of a product according to claim 1 or 2, characterized in that, The quorum sensing inhibitor A is used as a gargle, and after staying in the oral cavity for 10-30 s, it is spit out, and then the oral cavity is cleaned by using the spray B, thereby completing the oral disinfection operation for non-treatment purposes.

5. The method of antibacterial disinfection of the product according to claim 1 or 2, characterized in that, The spray A is used to clean the oral cavity, and after 30-60 s of action, the oral cavity is cleaned by using the spray B, thereby completing the oral disinfection operation for non-treatment purposes.

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