Method for synchronously disinfecting and sterilizing air and object surface by using peracetic acid composite disinfectant

Abstract

The invention discloses a method for synchronously disinfecting and sterilizing air and object surfaces by using a peracetic acid composite disinfectant, and aims to solve the problem that the disinfection effect and safety of peracetic acid are influenced by the influence of the temperature of a contact area or the surface when the peracetic acid is used for disinfecting indoor air and object surfaces. The disinfection and sterilization method comprises the following steps: 1, mixing hydrogen peroxide, glacial acetic acid, 1-butyl-3-methylimidazole phosphotungstate, 1-hydroxyethylidene-1, 1-diphosphonic acid and sodium glucoheptonate, stirring to react, and standing to obtain a high-concentration peracetic acid stock solution; 2, diluting the peracetic acid stock solution with standard hard water; and 3, filling the peracetic acid solution into a hydrogen peroxide aerial fog sterilizer or a vaporizing sterilizer, and sterilizing by using peracetic acid dry fog or vaporized peracetic acid. Vaporized peracetic acid not only has properties similar to those of vaporized hydrogen peroxide, but also enhances the disinfection effect and safety of the peracetic acid composite disinfectant by utilizing the synergistic effect of peracetic acid and hydrogen peroxide.

Description

Technical Field

[0001] The invention belongs to the field of disinfection, and in particular relates to an aerosol disinfection method using a composite peracetic acid disinfectant which can be used for simultaneous disinfection of air and the surface of an object. Background Art

[0002] Peracetic acid is known as an ideal broad-spectrum antimicrobial agent. It has high disinfection and sterilization capabilities, no harmful decomposition products, higher oxidation potential than hydrogen peroxide, and lacks catalase and peroxidase that cause hydrogen peroxide to inactivate. Peracetic acid is a more effective antimicrobial agent than hydrogen peroxide. Even at low concentrations and low temperatures, and in the presence of organic matter, it can still maintain effective disinfection capabilities against spores. Under the same sterilization conditions, peracetic acid has better disinfection and sterilization effects than hydrogen peroxide; compared with vaporized hydrogen peroxide, vaporized peracetic acid is faster and has a larger disinfection area for animal laboratories, clean rooms, biosafety cabinets, and animal care. However, at present, most of the peracetic acid disinfection of indoor air and object surfaces still remains at constant spray, ultra-low volume spray, and heated fumigation. These methods are easily condensed due to the influence of the temperature of the contact area or surface, which affects their disinfection efficacy and safety, and will cause the loss of peracetic acid during continuous heating. Therefore, the existing peracetic acid spraying with ultra-low volume sprayer requires 100mg / m 3 Or heating fumigation at least 1000mg / m 3 In addition, this method is prone to peracetic acid spillage during fumigation, which is dangerous.

[0003] Compared with peracetic acid, vaporized hydrogen peroxide disinfection has matured. At present, there are two main ways for hydrogen peroxide to occur in commercially available hydrogen peroxide disinfection machines. One is the vaporized hydrogen peroxide sterilization technology, which uses flash evaporation technology to drip a high-concentration hydrogen peroxide solution (generally >30%) onto the surface of a heated smooth metal object to instantly vaporize it into particles of 2 to 6 μm, and then spray it into the environment for disinfection and sterilization; the other is the hydrogen peroxide "dry mist" disinfection and sterilization technology, which uses ultra-high-speed rotation to physically centrifuge the hydrogen peroxide solution to form a uniform spray. The hydrogen peroxide concentration used in this technology is generally around 5%, with a small amount used and low corrosiveness, and can quickly and efficiently kill bacteria, molds, viruses, etc. However, hydrogen peroxide air disinfection also has some defects, mainly due to the lack of penetration of hydrogen peroxide. The results show that after 30% hydrogen peroxide is flash-evaporated to produce 1000ppm vaporized hydrogen peroxide for 20 minutes, the killing logarithm of anthrax spores on smooth surfaces is greater than 6, while the killing logarithm of carpet and pine wood surfaces is only 3.0 and 3.7. The possible reason is that the spores deposited on non-porous smooth surfaces can fully contact with vaporized hydrogen peroxide, while the spores of porous materials such as carpet and pine wood can easily penetrate into the pores or deep inside. Hydrogen peroxide has insufficient penetration and cannot kill the spores that penetrate inside. In addition, vaporized hydrogen peroxide is more unstable than peracetic acid, and it will decompose quickly after the occurrence, and its residence time in the air is short. Summary of the invention

[0004] The purpose of the present invention is to solve the problem that peracetic acid is easily condensed when used to disinfect indoor air and object surfaces due to the temperature of the contact area or surface, thereby affecting its disinfection efficacy and safety, and to provide a safe, environmentally friendly and multi-scenario applicable disinfection method, namely, a method for using peracetic acid to simultaneously disinfect and sterilize air and object surfaces.

[0005] The method for simultaneous disinfection and sterilization of air and surfaces with a composite peracetic acid disinfectant of the present invention is implemented by the following steps:

[0006] 1. Mix hydrogen peroxide, glacial acetic acid, 1-butyl-3-methylimidazole phosphotungstate, hydroxyethylidene diphosphonic acid and sodium glucoheptonic acid, stir and react for 8 to 12 hours, then let stand for treatment, and filter to obtain a high-concentration peracetic acid stock solution;

[0007] 2. Use standard hard water to dilute the high-concentration peracetic acid stock solution into a peracetic acid solution with a concentration of 500mg / L to 3000mg / L;

[0008] 3. Load the peracetic acid solution into the hydrogen peroxide aerosol disinfector, control the generation rate to 10mL / min~20mL / min, the particle size of the peracetic acid spray is ≤5μm, and spray the generated peracetic acid dry mist or nano mist into the air or on the surface of objects for disinfection and sterilization.

[0009] The method for simultaneous disinfection and sterilization of air and surfaces with a composite peracetic acid disinfectant of the present invention is implemented by the following steps:

[0010] 1. Mix hydrogen peroxide, glacial acetic acid, 1-butyl-3-methylimidazole phosphotungstate, hydroxyethylidene diphosphonic acid and sodium glucoheptonic acid, stir and react for 8 to 12 hours, then let stand for treatment, and filter to obtain a high-concentration peracetic acid stock solution;

[0011] 2. Use standard hard water to dilute the high-concentration peracetic acid stock solution into a peracetic acid solution with a concentration of 20g / L to 100g / L;

[0012] 3. Load the peracetic acid solution into the hydrogen peroxide vaporization disinfector, control the flash temperature to 110℃~150℃, and blow the generated vaporized peracetic acid into the air or onto the surface of objects through a fan for disinfection and sterilization.

[0013] The vaporized peracetic acid of the present invention not only has properties similar to those of vaporized hydrogen peroxide, but also utilizes the synergistic effect of peracetic acid and hydrogen peroxide to enhance the gas permeability sterilization ability of vaporized peracetic acid in narrow spaces and cavities similar to porous rocks through a fluid slip mechanism. Therefore, in closed spaces such as high-level biosafety laboratories, high-risk infectious disease foci, biosafety event sites, clean laboratories, clean workshops, hospitals, and logistics systems, hydrogen peroxide disinfection and sterilization technology is directly used to vaporize peracetic acid. Due to the use of a lower sterilization concentration and an improved vaporization process, as the half-life of staying on the surface is shortened, the exposure risk is reduced, and the compatibility problem with various materials is solved, it is suitable for various occasions as a safe and environmentally friendly low-temperature sterilization method. DETAILED DESCRIPTION

[0014] Specific implementation method 1: The method for simultaneous disinfection and sterilization of air and surfaces using a peracetic acid composite disinfectant in this implementation method is implemented according to the following steps:

[0015] 1. Mix hydrogen peroxide, glacial acetic acid, 1-butyl-3-methylimidazole phosphotungstate, hydroxyethylidene diphosphonic acid and sodium glucoheptonic acid, stir and react for 8 to 12 hours, then let stand for treatment, and filter to obtain a high-concentration peracetic acid stock solution;

[0016] 2. Use standard hard water to dilute the high-concentration peracetic acid stock solution into a peracetic acid solution with a concentration of 500mg / L to 3000mg / L;

[0017] 3. Load the peracetic acid solution into the hydrogen peroxide aerosol disinfector, control the generation rate to 10mL / min~20mL / min, the particle size of the peracetic acid spray is ≤5μm, and spray the generated peracetic acid dry mist or nano mist into the air or on the surface of objects for disinfection and sterilization.

[0018] This embodiment uses hydrogen peroxide "dry mist" and "nano mist" disinfection and sterilization technology to produce peracetic acid.

[0019] This embodiment is used in closed spaces such as high-level biosafety laboratories, high-risk infectious disease foci, biosafety incident sites, clean laboratories, clean workshops, hospitals, and logistics systems to kill bacterial spores, viruses, bacterial propagules, and fungi in the air and on the surface of objects using low-concentration peracetic acid aerosol, and achieves a sterilization effect. The bacterial spores are selected from at least one of the following: anthrax spores, Bacillus subtilis var. niger spores, Bacillus stearothermophilus spores, and Bacillus cereus spores;

[0020] The virus comprises an enveloped virus and a non-enveloped virus, and is selected from at least one of the following: polio virus, HIV, mad cow virus, Creutzfeldt-Jakob virus, new coronavirus, hand, foot and mouth disease virus, avian influenza virus, influenza A (H1N1) virus and influenza A (H7N9) virus;

[0021] The bacteria and fungi are selected from at least one of the following: intestinal pathogens, pyogenic cocci, pathogenic yeasts, common pathogens of hospital infections and drug-resistant bacteria.

[0022] Specific embodiment 2: This embodiment is different from the specific embodiment 1 in that the preparation method of 1-butyl-3-methylimidazolium phosphotungstate in step 1 is as follows:

[0023] 0.01 mol of phosphotungstic acid was dissolved in 100 mL of water to prepare a phosphotungstic acid aqueous solution, and then 0.06 mol of 1-butyl-3-methylimidazole bromide [Bmim] Br was added to the phosphotungstic acid aqueous solution, and the reaction was stirred at room temperature to collect the reaction precipitate, and 1-butyl-3-methylimidazole phosphotungstate was obtained after washing and drying.

[0024] 1-Butyl-3-methylimidazolium phosphotungstate [Bmim] prepared in this embodiment 3 PW 12 O 40 It is a white powder and is used as a catalyst for preparing peracetic acid stock solution.

[0025] Specific implementation method three: This implementation method is different from specific implementation method two in that the stirring reaction time at room temperature is 10 to 15 hours.

[0026] Specific embodiment 4: This embodiment is different from specific embodiments 1 to 3 in that in step 1, 300 to 350 mL of hydrogen peroxide with a mass fraction of 30%, 600 to 700 mL of glacial acetic acid, 0.03 to 0.08 g of 1-butyl-3-methylimidazole phosphotungstate, 3 to 3.5 g of hydroxyethylidene diphosphonic acid and 1 to 2 g of sodium glucoheptonic acid are mixed, stirred for reaction for 8 to 12 hours, and then allowed to stand for treatment, and filtered to obtain a high concentration peracetic acid stock solution.

[0027] Specific implementation method 5: This implementation method is different from specific implementation method 4 in that the static treatment time is 4 to 6 days.

[0028] Specific embodiment 6: This embodiment is different from specific embodiments 1 to 5 in that the concentration of peracetic acid in the high concentration peracetic acid stock solution in step 1 is 80 to 120 g / L, and the concentration of hydrogen peroxide is 40 to 70 g / L.

[0029] Specific embodiment 7: This embodiment is different from the specific embodiments 1 to 6 in that the amount of peracetic acid generated in step 3 is controlled to be 10 mg / m 3 ~30 mg / m 3 The action time is 30min to 120min.

[0030] In this embodiment, the relative humidity (RH) of the air is 60% to 80%.

[0031] Specific implementation method eight: The method for simultaneous disinfection and sterilization of air and surfaces using a composite peracetic acid disinfectant in this implementation method is implemented according to the following steps:

[0032] 1. Mix hydrogen peroxide, glacial acetic acid, 1-butyl-3-methylimidazole phosphotungstate, hydroxyethylidene diphosphonic acid and sodium glucoheptonic acid, stir and react for 8 to 12 hours, then let stand for treatment, and filter to obtain a high-concentration peracetic acid stock solution;

[0033] 2. Use standard hard water to dilute the high-concentration peracetic acid stock solution into a peracetic acid solution with a concentration of 20g / L to 100g / L;

[0034] 3. Load the peracetic acid solution into the hydrogen peroxide vaporization disinfector, control the flash temperature to 110℃~150℃, and blow the generated vaporized peracetic acid into the air or onto the surface of objects through a fan for disinfection and sterilization.

[0035] This embodiment uses hydrogen peroxide "vaporization" disinfection and sterilization technology (flash technology) to generate peracetic acid.

[0036] Specific embodiment 9: This embodiment is different from specific embodiment 8 in that the generation rate of vaporized peracetic acid in step 3 is controlled to be 0.8 mL / min to 1.2 mL / min, and the generation amount is 80 mg / m3 ~120 mg / m 3 .

[0037] In this embodiment, the amount of generation is per 1m 3 The amount of vaporized peracetic acid sprayed into the space is 80 to 120 mg.

[0038] Specific implementation method ten: The difference between this implementation method and specific implementation method eight is that the disinfection and sterilization time in step three is 30 minutes to 120 minutes.

[0039] Embodiment: The method for simultaneous disinfection and sterilization of air and surfaces using a composite peracetic acid disinfectant in this embodiment is implemented according to the following steps:

[0040] 1. Dissolve phosphotungstic acid (0.01mol, 28.8g) in 100ml water to prepare phosphotungstic acid aqueous solution, then add 1-butyl-3-methylimidazole bromide [Bmim]Br (0.06mol, 13.2g) to the phosphotungstic acid aqueous solution, control the stirring speed to about 1000 rpm, and keep observing and reduce the stirring speed in time within 2h during the stirring process to reduce splashing. Stir at room temperature for 12h to obtain a white precipitate, filter and wash with water for more than 3 times until there is no bromide ion in the filtrate, and dry the obtained solid at 80℃ for 24h to obtain a white powder [Bmim] 3 PW 12 O 40 catalyst;

[0041] 2. Place 333 mL of hydrogen peroxide (30%), 666 mL of glacial acetic acid, 0.05 g of 1-butyl-3-methylimidazolium phosphotungstate, 3.333 g of hydroxyethylidene diphosphonic acid and 1.666 g of sodium glucoheptonic acid in a three-necked flask, stir magnetically for 10 h, transfer to a glass bottle, let stand for 5 days and then filter to obtain a high-concentration peracetic acid stock solution that can be stored for a long time (peracetic acid concentration>100 g / L, hydrogen peroxide concentration is 40 g / L);

[0042] 3. Use standard hard water to dilute the high concentration peracetic acid stock solution into peracetic acid solutions with concentrations of 1000 mg / L, 2000 mg / L, and 3000 mg / L (calculated as peracetic acid).

[0043] Reagents and materials used in this example:

[0044] Test strain: Bacillus subtilis var. niger (ATCC 9372) spores.

[0045] DE Neutralizing Broth: Weigh 39.02 g and add to 1000 mL of distilled water, divide into portions, sterilize with 121°C pressure steam for 15 min, and then conduct a neutralizing agent identification test. After passing the identification test, use it for later use;

[0046] Tryptone soy broth (TSB): weigh 30 g and add into 1000 mL distilled water, sterilize by pressure steam at 121°C for 15 min before use;

[0047] Tryptone soy agar (TSA): weigh 40 g and add to 1000 mL of distilled water. Sterilize with pressure steam at 121°C for 15 min before use.

[0048] Diluent (TPS): 1 g / L tryptone and 8.5 g / L sodium chloride were added to 1000 mL of distilled water and sterilized by pressure steam at 121°C for 15 min before use;

[0049] Nutrient broth (NB) medium: weigh 18 g and add into 1000 mL distilled water, sterilize by pressure steam at 121°C for 15 min before use;

[0050] Test tube: 1.8cm×18cm, sterilized for later use;

[0051] Carrier: White plain cotton cloth, which should be degreased first. The degreasing method is as follows: ① Boil the carrier in water containing detergent for 30 minutes; ② Wash with tap water; ③ Boil with distilled water for 10 minutes; ④ Rinse with distilled water until the pH is neutral; ⑤ Dry and iron for use. Then, remove one warp and one weft yarn around the edge of the degreased cloth according to the size specified by the carrier, and then cut it into carriers of 10mm×10mm according to the yarn drawing marks, and sterilize it with pressure steam for use.

[0052] Aerosol chamber: A pair of adjacent aerosol chambers are constructed with stainless steel and glass, one for disinfection experiments and the other for experimental controls. Both are equipped with temperature and humidity control devices, as well as ventilators, filtration sterilization or other disinfection devices and corresponding pipelines, as well as windows for opening spray infection, disinfectant administration, sampling, sleeve operation, sample transfer and air stirring devices.

[0053] The test grouping of this embodiment is as follows:

[0054] Test group: Bacillus subtilis black variant (ATCC 9372) spore-stained slides, peracetic acid generation (measured in peracetic acid) was 10 mg / m 3 , 20mg / m 3 , 30mg / m 3 , the test was carried out at action time of 30min, 60min, 90min and 120min;

[0055] Positive control group: Use standard hard water instead of disinfectant solution and conduct the test according to the same steps as above. The result represents the bacterial content in the test system and is used as the live bacterial concentration of the control group;

[0056] Negative control group: observe whether the relevant reagents and culture medium used in the same experiment are contaminated.

[0057] Application Example 1: The peracetic acid solution is loaded into a hydrogen peroxide aerosol disinfector, and the generated peracetic acid spray is sprayed onto the surface of an object for disinfection and sterilization.

[0058] The simulated field experiment procedure is as follows:

[0059] Adjust 20m 3 The temperature in the aerosol chamber is 20±1℃. Place the pre-prepared bacterial slices in a sterile plate in the most difficult to sterilize position in the aerosol chamber. Set the flow rate of the hydrogen peroxide aerosol disinfector to 20mL / min, add 200mL of the prepared 1000mg / L, 2000mg / L, and 3000mg / L peracetic acid solution, place the hydrogen peroxide aerosol disinfector in a corner of the aerosol chamber, and start the disinfector. After the occurrence is over, the humidity in the aerosol chamber is about 70%.

[0060] Start the disinfection timer when the disinfection machine is turned on. Take out a bacterial slice at 30, 60, 90, and 120 minutes after the start of disinfection and place it in a test tube containing 5 mL of DE neutralization broth. After shaking for 40 seconds or tapping 200 times, take 1.0 mL of the eluate and inoculate it on a plate. Inoculate two plates for each sample.

[0061] Preparation of the bacterial staining sheet: Spread the sterilized carrier sheet flat in a sterile plate, drip spore solution of Bacillus subtilis black variant (ATCC 9372) on each sheet, with a dripping amount of 10 μL per sheet. After dripping, the bacterial staining carrier can be placed in a 37°C incubator to dry (about 20 minutes to 30 minutes), or placed in the shade at room temperature to dry naturally before use. The number of recovered colonies on each bacterial sheet is 1×10 6 CFU / piece~5×10 6 CFU / tablet.

[0062] For the positive control group, the control group aerosol chamber was treated with standard hard water instead of disinfectant. After the treatment of the test group was completed, the infected carrier was placed in a test tube containing 5 mL of neutralizer solution, shaken for 40 seconds or knocked 200 times, and diluted 10 times with diluent. The suspension with appropriate dilution was selected, and 1.0 mL was drawn to inoculate the plate, and two plates were inoculated for each sample. At the same time, 1.0 mL of the neutralizer and diluent for the test were drawn to inoculate the plate, and two plates were inoculated for each sample as the negative control group. After each group was inoculated with the plate, 15 mL to 20 mL of TSA was poured, and after solidification, it was placed in a 36°C ± 1°C incubator for 72 hours, and the number of colonies was counted. The test was repeated 3 times.

[0063] The experimental results are as follows:

[0064] Use hydrogen peroxide "dry fog" and "nano fog" disinfection and sterilization technology. When the flow rate of the disinfection machine is 20mL / min, add 200mL of 1000mg / L, 2000mg / L, and 3000mg / L peracetic acid solution to generate peracetic acid aerosol to 20m 3 The black variant of Bacillus subtilis spores were disinfected in the aerosol chamber. As shown in Table 1, when the amount of peracetic acid was 10 mg / m 3 When the peracetic acid amount is 20mg / m 3 When the peracetic acid amount is 30mg / m 3 The sterilization effect of Bacillus subtilis var. niger spores on the cloth piece can be achieved within 30 minutes (killing logarithm>6.00).

[0065] Table 1 The disinfection effect of hydrogen peroxide "dry fog" and "nano fog" disinfection and sterilization technology on the surface of objects by generating peracetic acid

[0066]

[0067]

[0068] Application Example 2: The peracetic acid solution is loaded into a hydrogen peroxide vaporization disinfection machine, and the generated vaporized peracetic acid is sprayed onto the surface of the object for disinfection and sterilization.

[0069] The simulated field experiment procedure of the hydrogen peroxide "vaporization" disinfection and sterilization technology (flash evaporation technology) to generate peracetic acid on the surface of an object is as follows:

[0070] Adjust 20m 3 The temperature in the aerosol chamber is 20±1℃, and the RH is 60%-70%. The pre-prepared bacterial slices are placed in the most difficult to sterilize position in the aerosol chamber using sterile plates. The flash temperature of the hydrogen peroxide "vaporization" sterilizer is set to 120℃, the flow rate is 1.0mL / min, and the prepared 100g / L peracetic acid solution is added. The peracetic acid generation time is set to 12min, 18min, and 24min (equivalent to a peracetic acid generation amount of 60mg / m 3 , 90mg / m 3 , 120mg / m 3), place the hydrogen peroxide vaporizing disinfection machine in the center of the aerosol chamber and start the disinfection machine.

[0071] Start the disinfection timer when the disinfection machine is turned on. Take out a bacterial slice at 30, 60, 90, and 120 minutes after the start of disinfection and place it in a test tube containing 5 mL of DE neutralization broth. After shaking for 40 seconds or tapping 200 times, take 1.0 mL of the eluate and inoculate it on a plate. Inoculate two plates for each sample.

[0072] For the positive control group, the control group aerosol chamber was treated with standard hard water instead of disinfectant. After the treatment of the test group was completed, the infected carrier was placed in a test tube containing 5 mL of neutralizer solution, shaken for 40 seconds or knocked 200 times, and diluted 10 times with diluent. The suspension with appropriate dilution was selected, and 1.0 mL was drawn to inoculate the plate, and two plates were inoculated for each sample. At the same time, 1.0 mL of the neutralizer and diluent for the test were drawn to inoculate the plate, and two plates were inoculated for each sample as the negative control group. After each group was inoculated with the plate, 15 mL to 20 mL of TSA was poured in, and after solidification, it was placed in a 36°C ± 1°C incubator for 72 hours, and the number of colonies was counted. The test was repeated 3 times.

[0073] The experimental results are as follows:

[0074] Under the conditions of aerosol room temperature of 20±1℃ and RH of 60%~70%, hydrogen peroxide "vaporization" disinfection and sterilization technology (flash technology) was used, and the flash temperature of hydrogen peroxide "vaporization" disinfection machine was set to 120℃ and the flow rate was 1.0mL / min. 100g / L peracetic acid was added and the use time was 12min, 18min, and 24min respectively (equivalent to 60mg / m 3 , 90mg / m 3 , 120mg / m 3 ), for 20m 3 The black variant of Bacillus subtilis spores were disinfected in the aerosol chamber. As shown in Table 2, when the amount of peracetic acid was 60 mg / m 3 When the peracetic acid generation amount is 90mg / m 3 When the peracetic acid generation amount is 120mg / m 3The sterilization effect of Bacillus subtilis var. niger spores on the cloth piece can be achieved within 30 minutes (killing logarithm>6.00).

[0075] Table 2 The disinfection effect of hydrogen peroxide "vaporization" disinfection and sterilization technology (flash technology) on the surface of objects using peracetic acid

[0076]

[0077] Application Example 3: The peracetic acid solution is loaded into a hydrogen peroxide aerosol disinfector, and the generated peracetic acid mist is sprayed into the air for disinfection and sterilization.

[0078] Test 1: Neutralizer Identification Test

[0079] Test strain: Staphylococcus albuterol (8032).

[0080] Organic interferences: 3.0% bovine serum albumin;

[0081] Tryptone soy agar (TSA): weigh 40 g and add to 1000 mL of distilled water. Sterilize with pressure steam at 121°C for 15 min before use.

[0082] Diluent (TPS): 1 g / L tryptone and 8.5 g / L sodium chloride were added to 1000 mL of distilled water and sterilized by pressure steam at 121°C for 15 min before use;

[0083] Nutrient agar medium (containing neutralizer): weigh 33 g of nutrient agar and 0.05 g of sodium thiosulfate and add them to 1000 mL of distilled water. Sterilize with pressure steam at 121 °C for 15 min before use.

[0084] Test tube: 1.8cm×18cm, sterilized for later use;

[0085] Aerosol chamber: A pair of adjacent aerosol chambers are constructed with stainless steel and glass, one for disinfection experiments and the other for experimental controls. Both are equipped with temperature and humidity control devices, as well as ventilators, filtration sterilization or other disinfection devices and corresponding pipelines, as well as windows for opening spray infection, disinfectant administration, sampling, sleeve operation, sample transfer and air stirring devices.

[0086] Bacteria infection device: air compressor, pressure gauge, gas flow meter, aerosol generator;

[0087] Air microorganism sampling device: six-stage sieve air impact sampler, air extraction equipment;

[0088] Environmental monitoring equipment: temperature and humidity meter.

[0089] The experimental operation process of this embodiment is as follows:

[0090] (1) Preparation of bacterial suspension: Take 18-24 h fresh culture of 3-8 generations of Staphylococcus aureus, repeatedly blow and wash with 5 mL of nutrient broth medium to wash off the bacterial moss, and dilute with nutrient broth to a concentration of 5 × 10 2 CFU / mL~3×10 3 CFU / mL, for future use.

[0091] (2) Experimental groups and operation steps

[0092] Group 1: Take 0.1 mL of the diluted bacterial suspension and spread it evenly on two nutrient agar plates containing neutralizers to count the live bacteria. Observe whether the neutralizer has an inhibitory effect on the growth of the test bacteria;

[0093] Group 2: Set the flow rate of the hydrogen peroxide aerosol disinfector to 20mL / min, add 200mL of the prepared 1000mg / L peracetic acid solution and place it in a corner of the aerosol chamber, start the disinfector, let it work for 30min, and immediately use a six-stage air impact sampler equipped with a nutrient agar plate containing a neutralizer to sample. The air sampling volume is the same as the preset sampling time (or sampling volume) after disinfection in the air disinfection identification test. After the sampling is completed, let it work for 10min, and then take 0.1mL of the above-mentioned diluted bacterial suspension respectively, and evenly spread it on 6 plates where the disinfectant was collected, and count the live bacteria culture to observe whether the neutralization product has an inhibitory effect on the growth of the test bacteria.

[0094] Group 3: Take 0.1 mL of the diluted bacterial suspension and spread it evenly on the surface of 2 ordinary nutrient agar plates to count the live bacteria as a bacterial count control.

[0095] Group 4: Pipette 0.1 mL of the diluent respectively, spread it evenly on two nutrient agar plates containing a neutralizer and two ordinary nutrient agar plates, and culture them in a constant temperature incubator at 36℃±1℃ for 48 h, as the negative control group.

[0096] (3) Evaluation regulations: If the test results meet all the following conditions, the tested neutralizer can be judged as qualified. The bacterial count of Groups 1 to 3 is 50 CFU / plate to 300 CFU / plate, and the error rate of colony counts among the three groups should not exceed 15%; Group 4 has sterile growth; The test is repeated 3 times, and each test should meet the above conditions.

[0097] The test results are as follows:

[0098] As shown in Table 3, the test was repeated 3 times, and the colony counts of all plates from Group 1 to Group 3 were 74-95 CFU / plate. The colony count error rate among the three groups should be 2.96% to 5.06%, not exceeding 15%. Group 4 showed aseptic growth, and the test results of the four groups all met the standard requirements. This indicates that the selected neutralizer can effectively neutralize the peracetic acid compound disinfectant, and the neutralizer and the neutralization product solution have no effect on the growth of Bacillus subtilis variant spores.

[0099] Table 3 Neutralizer identification test results

[0100]

[0101]

[0102] Experiment 2: Evaluation of air disinfection effect

[0103] 1. Reagents and Materials Used

[0104] Test strain: Staphylococcus albuterol (8032).

[0105] Organic interferences: 3.0% bovine serum albumin;

[0106] Tryptone soy agar (TSA): weigh 40 g and add to 1000 mL of distilled water. Sterilize with pressure steam at 121°C for 15 min before use.

[0107] Diluent (TPS): 1 g / L tryptone and 8.5 g / L sodium chloride were added to 1000 mL of distilled water and sterilized by pressure steam at 121°C for 15 min before use;

[0108] Nutrient agar medium (containing neutralizer): weigh 33 g of nutrient agar and 0.05 g of sodium thiosulfate and add them to 1000 mL of distilled water. Sterilize with pressure steam at 121 °C for 15 min before use.

[0109] Anti-foaming agent: olive oil;

[0110] Test tube: 1.8cm×18cm, sterilized for later use;

[0111] Aerosol chamber: A pair of adjacent aerosol chambers are constructed with stainless steel and glass, one for disinfection experiments and the other for experimental controls. Both are equipped with temperature and humidity control devices, as well as ventilators, filtration sterilization or other disinfection devices and corresponding pipelines, as well as windows for opening spray infection, disinfectant administration, sampling, sleeve operation, sample transfer and air stirring devices.

[0112] Bacteria infection device: air compressor, pressure gauge, gas flow meter, aerosol generator;

[0113] Air microorganism sampling device: six-stage sieve air impact sampler, air extraction equipment;

[0114] Environmental monitoring equipment: temperature and humidity meter.

[0115] 2. Test operation

[0116] (1) Preparation of bacterial suspension: Take 18-24 h fresh culture of 3-8 generations of Staphylococcus aureus, repeatedly blow and wash with 5 mL of nutrient broth medium, wash off the bacterial moss, filter with sterile absorbent cotton, and dilute with nutrient broth to a concentration of 5 × 10 7 CFU / mL~5×10 8 CFU / mL, for future use.

[0117] (2) Experimental groups and operation steps

[0118] At the same time, adjust the temperature of the two aerosol chambers (20±1℃). Put the equipment to be used into the aerosol chamber at once and close the door. After that, all operations and instrument control are carried out outdoors through a window with a sealed sleeve or a remote control. The door can be opened only after the test is completed.

[0119] The two aerosol chambers were sprayed with bacteria at the same time according to the set pressure, gas flow (28.3L / s) and spray time (5min); the spray was stirred with a fan; after the spray was completed, the stirring was continued for 5min, and then it was left to stand for 5min. At the same time, the control group and the test group were sampled before disinfection using a six-stage sieve air impact sampler, which served as the positive control (i.e., the amount of contaminated bacteria) before the start of the control group experiment and before the disinfection of the test group.

[0120] Set the flow rate of hydrogen peroxide mist disinfector to 20mL / min, add 200mL of prepared 1000mg / L peracetic acid solution (equivalent to the theoretical peracetic acid concentration of 10mg / m 3 ), place the hydrogen peroxide aerosol disinfection machine in a corner of the aerosol chamber and start the disinfection machine. After the occurrence is over, the humidity in the aerosol chamber is about 70%.

[0121] Start the disinfection timer when the disinfection machine is turned on. Take samples of the test group and the control group at the same time according to the above method 30 and 60 minutes after the start of disinfection. Put the sample plate directly into the 37℃ incubator and incubate for 48 hours. Observe the final results and count the number of growing colonies.

[0122] After completing the sampling and sample inoculation of the test group and the positive control group, the unused culture medium, sampling solution and PBS (1 to 2 portions each) from the same batch should be cultured or inoculated with the above two groups of samples as negative controls. If bacteria grow in the negative control group, it means that the culture medium or reagents used are contaminated, the test is invalid, and the sterile equipment should be replaced and repeated.

[0123] 3. Evaluation Method

[0124] The same test conditions are repeated 3 times. When the killing rate of the 3 results is ≥99.90%, the disinfection is considered qualified. The calculation method of killing rate is as follows:

[0125]

[0126] N t : The natural extinction rate of bacteria in the air;

[0127] V 0 With V t : are the air bacteria content before and at different times during the control group experiment;

[0128] K t : The killing rate of airborne bacteria by disinfection treatment;

[0129] V 0 'With V t ': respectively represents the bacterial content in the air before and at different times during the disinfection process of the experimental group.

[0130] The bacterial content in the air before and after disinfection is calculated according to the following formula:

[0131]

[0132] 4. Experimental Results

[0133] Use hydrogen peroxide "dry fog" and "nano fog" disinfection and sterilization technology. When the flow rate of the disinfection machine is 20mL / min, add 200mL of 1000mg / L peracetic acid solution to generate peracetic acid aerosol to 20m 3 The white Staphylococcus aureus in the aerosol chamber was disinfected. It can be seen from Table 4 that when the amount of peracetic acid was 10 mg / m 3 At this time, the sterilization rate of white Staphylococcus in the air of the aerosol chamber was 100.00%. At this time, the natural decay rate of white Staphylococcus in the air of the control group was below 26.44%.

[0134] Table 4 The killing effect of hydrogen peroxide "dry fog" and "nano fog" disinfection and sterilization technology on Staphylococcus aureus by generating peracetic acid

[0135]

[0136] Application Example 4: The peracetic acid solution is loaded into a hydrogen peroxide vaporization disinfection machine, and the generated vaporized peracetic acid is sprayed into the air for disinfection and sterilization.

[0137] The experimental operation process of this embodiment is as follows:

[0138] (1) Preparation of bacterial suspension: Take 18-24 h fresh culture of 3-8 generations of Staphylococcus aureus, repeatedly blow and wash with 5 mL of nutrient broth medium, wash off the bacterial moss, filter with sterile absorbent cotton, and dilute with nutrient broth to a concentration of 5 × 10 7 CFU / mL~5×10 8 CFU / mL, for future use.

[0139] (2) Experimental groups and operation steps

[0140] At the same time, adjust the temperature of the two aerosol chambers (20±1℃). Put the equipment to be used into the aerosol chamber at once and close the door. After that, all operations and instrument control are carried out outdoors through a window with a sealed sleeve or a remote control. The door can be opened only after the test is completed.

[0141] The two aerosol chambers were sprayed with bacteria at the same time according to the set pressure, gas flow (28.3L / s) and spray time (5min); the spray was stirred with a fan; after the spray was completed, the stirring was continued for 5min, and then it was left to stand for 5min. At the same time, the control group and the test group were sampled before disinfection using a six-stage sieve air impact sampler, which served as the positive control (i.e., the amount of contaminated bacteria) before the start of the control group experiment and before the disinfection of the test group.

[0142] Set the flash temperature of the hydrogen peroxide "vaporization" disinfection machine to 120°C and the flow rate to 5.0 mL / min, add the prepared 2000 mg / L peracetic acid solution, and set the peracetic acid generation time to 20 minutes (equivalent to a peracetic acid generation amount of 10 mg / m 3 ), place the hydrogen peroxide "vaporization" disinfection machine in the center of the aerosol chamber and start the disinfection machine.

[0143] Start the disinfection timer when the disinfection machine is turned on. Take samples of the test group and the control group at the same time according to the above method 30 and 60 minutes after the start of disinfection. Put the sample plate directly into the 37℃ incubator and incubate for 48 hours. Observe the final results and count the number of growing colonies.

[0144] After completing the sampling and sample inoculation of the test group and the positive control group, the unused culture medium, sampling solution and PBS (1 to 2 portions each) from the same batch should be cultured or inoculated with the above two groups of samples as negative controls. If bacteria grow in the negative control group, it means that the culture medium or reagents used are contaminated, the test is invalid, and the sterile equipment should be replaced and repeated.

[0145] The experimental results are as follows:

[0146] Under the conditions of 20±1℃ and RH 60%~70% in the aerosol room, hydrogen peroxide "vaporization" disinfection and sterilization technology (flash technology) was used to sterilize 20m3 The white Staphylococcus aureus in the aerosol chamber was disinfected. It can be seen from Table 5 that when the amount of peracetic acid was 10 mg / m 3 When the disinfection lasted for 30 minutes, the sterilization rate of Staphylococcus aureus in the air of the aerosol chamber was 100.00%. At this time, the natural decay rate of Staphylococcus aureus in the air of the control group was below 20.30%.

[0147] Table 5 The killing effect of peracetic acid on Staphylococcus aureus by hydrogen peroxide "vaporization" disinfection and sterilization technology (flash technology)

[0148]

[0149] Application Example 5: The metal corrosivity of the disinfection method described in Application Example 2 was measured.

[0150] The disinfection method described in this embodiment was tested for metal corrosion according to GB / T 38498-2020.

[0151] 1. Materials used

[0152] Metal sheet: round, diameter 24±0.1mm, thickness 1.0mm, center hole diameter 2.0mm, total surface area approximately 9.80cm 2 . The finish is 4 to 6. The raw materials are as follows: carbon steel (GB700-65), stainless steel (GB1220-75), copper (GB2060-80), aluminum (GB1173-74).

[0153] Sandpaper: 120 grit water-repellent sandpaper, GB 2477.

[0154] Weighing cup.

[0155] Balance: Sensitivity 0.1mg.

[0156] Aerosol chamber: The aerosol chamber is constructed of stainless steel and glass, one for disinfection experiments and the other for experimental control. Both are equipped with temperature and humidity control devices, as well as ventilator filtration sterilization or other disinfection devices and corresponding pipelines, as well as windows for opening spray infection, disinfectant administration, sampling, sleeve operation, sample transfer and air stirring devices.

[0157] Hydrogen peroxide "vaporization" disinfection machine: using flash evaporation technology, more than 90% of the aerosol particles sprayed out are between 1μm and 10μm.

[0158] 2. Experimental Procedure

[0159] (1) Treatment of metal sheets before the experiment: First, soak 3 pieces of each of the four metal sheets in a surfactant-containing detergent for 10 minutes, thoroughly degrease, and wash; use 120-grit sandpaper to remove the oxide layer on both sides and the surrounding surface of the metal sheet, and then rinse with tap water. Degrease again with anhydrous ethanol, dry in a 50°C constant temperature box for 1 hour, and weigh after the temperature drops to room temperature. After the balance returns to zero, weigh each metal sheet 3 times, accurate to 0.1 mg, and take the average value as the weight before the test. When weighing, wear clean gloves and do not touch the sample directly with your hands.

[0160] (2) Placement of metal samples: 3 pieces of each metal sample are hung in order along one diagonal side of the aerosol chamber at equal distances from the inside, middle and outside, 0.8m to 1.2m above the ground. The placement and position of the test group and the control group should be the same. The test surface of the metal sample should not be directly exposed to the spray.

[0161] (3) Cyclic test: adjust two 20m 3 The temperature of the aerosol chamber was 20℃~25℃, and the relative humidity was 70%~80%. The experimental group used hydrogen peroxide "vaporization" disinfection and sterilization technology (flash technology), and the flash temperature of the hydrogen peroxide "vaporization" disinfection machine was set to 120℃, the flow rate was 1.0mL / min, and 100g / L peracetic acid was added for 24min (equivalent to 120mg / m 3 ), the disinfection time is calculated from the start of the machine, 120 minutes of disinfection and 30 minutes of ventilation after disinfection are recorded as one cycle, and 45 cycles are performed on the suspended metal samples.

[0162] (4) Sample processing: After the cycle is completed, take out the metal sample, rinse it with tap water, and then use a brush or other soft tools to remove the corrosion products; after the metal sample is cleaned and the corrosion products are removed, use coarse filter paper to absorb the moisture, place it in a plate with filter paper, put it in a 50℃ incubator, dry it for 1 hour, pick it up with tweezers, and weigh it on the balance after the temperature drops to room temperature; weigh it three times after the balance returns to zero, and take the average value as the weight after the test. When weighing, wear clean gloves as before the test, and do not touch the sample directly with your hands (the same below).

[0163] During the entire test period, the test shall not be interrupted. When the test must be interrupted for a long time, the metal samples of the test group and the control group should be taken out of the aerosol chamber at the same time, and the metal samples should be treated in the same way as the metal samples after the test is completed, and stored in the desiccator until the test is resumed.

[0164] The control group used pure water instead of disinfectant or the disinfection factor of the disinfection machine was turned off. The rest of the test steps and processes were the same as those of the test group. After the cycle was completed, the metal samples were taken out and cleaned, chemically treated, rinsed with water, dried, and weighed in the same way as the metal samples in the test group, and their average weight loss was calculated.

[0165] (5) Result calculation and evaluation criteria:

[0166] Observe and record the color change of the metal sheet and express it as the average value of the metal corrosion rate (R). When calculating, the weight loss value of the blank control group sample should be subtracted. The calculation formula is as follows

[0167]

[0168] [R is the corrosion rate, mm / a (millimeter / year); m is the weight of the metal sheet before the test, g; m t is the weight of the metal sheet after the test, g; m k The weight loss value of the sample after chemical treatment to remove corrosion products, g. If no chemical removal treatment is performed in the test, m is deleted from the formula during calculation. k Value; S is the total surface area of ​​the metal sheet, cm 2 ; t is the test time, h; d is the density of the metal material, kg / m 3 ].

[0169] Corrosivity classification standards:

[0170] Table 6 Corrosivity classification standards

[0171] Corrosion rate R(mm / a) level <0.0100 Almost no corrosion 0.0100～<0.100 Mild corrosion 0.100～<1.00 Moderate corrosion ≥1.00 Severe corrosion

[0172] 4. Experimental Results

[0173] Table 7 Corrosion degree of metals by application example 2 of the present invention

[0174]

[0175] At room temperature of 20℃~25℃ and relative humidity of 70%~80%, use hydrogen peroxide "vaporization" disinfection and sterilization technology (flash technology), set the hydrogen peroxide "vaporization" disinfection machine flash temperature to 120℃ and flow rate to 1.0mL / min, add 100g / L peracetic acid solution for 24min (equivalent to 120mg / m 3), the disinfection time is calculated from the start of the machine, 120 minutes of disinfection and 30 minutes of ventilation after disinfection are recorded as one cycle, and the suspended metal samples are cycled 45 times. As shown in Table 7, the corrosion rate of the hydrogen peroxide "vaporization" disinfection and sterilization technology (flash technology) with peracetic acid in the experimental group and the control group on stainless steel is basically no corrosion, and the corrosion rate of the hydrogen peroxide "vaporization" disinfection and sterilization technology (flash technology) with peracetic acid in the experimental group and the control group on aluminum, carbon steel and copper is mild corrosion. The test results show that the corrosion is caused by the humidity formed by the spray, and is not entirely due to the vaporization of the disinfectant.

Claims

1. A method for simultaneous disinfection and sterilization of air and surfaces using a composite peracetic acid disinfectant, characterized in that The method for simultaneous disinfection and sterilization of air and surfaces using a peracetic acid composite disinfectant is achieved by the following steps:

1. Mix hydrogen peroxide, glacial acetic acid, 1-butyl-3-methylimidazole phosphotungstate, hydroxyethylidene diphosphonic acid and sodium glucoheptonic acid, stir and react for 8 to 12 hours, then let stand for treatment, and filter to obtain a high-concentration peracetic acid stock solution; 2. Use standard hard water to dilute the high-concentration peracetic acid stock solution into a peracetic acid solution with a concentration of 500mg / L to 3000mg / L; 3. Load the peracetic acid solution into the hydrogen peroxide aerosol disinfector, control the generation rate to 10mL / min~20mL / min, the particle size of the peracetic acid spray is ≤5μm, and spray the generated peracetic acid dry mist or nano mist into the air or on the surface of objects for disinfection and sterilization.

2. The method for simultaneous disinfection and sterilization of air and surfaces using a composite peracetic acid disinfectant according to claim 1, characterized in that The preparation method of 1-butyl-3-methylimidazolium phosphotungstate in step 1 is as follows: 0.01 mol of phosphotungstic acid was dissolved in 100 mL of water to prepare a phosphotungstic acid aqueous solution, and then 0.06 mol of 1-butyl-3-methylimidazole bromide [Bmim] Br was added to the phosphotungstic acid aqueous solution, and the reaction was stirred at room temperature to collect the reaction precipitate, and 1-butyl-3-methylimidazole phosphotungstate was obtained after washing and drying.

3. The method for simultaneous disinfection and sterilization of air and surfaces using a composite peracetic acid disinfectant according to claim 2, characterized in that The stirring reaction time at room temperature is 10 to 15 hours.

4. The method for simultaneous disinfection and sterilization of air and surfaces using a composite peracetic acid disinfectant according to claim 1, characterized in that In step 1, 300-350 mL of 30% hydrogen peroxide, 600-700 mL of glacial acetic acid, 0.03-0.08 g of 1-butyl-3-methylimidazole phosphotungstate, 3-3.5 g of hydroxyethylidene diphosphonic acid and 1-2 g of sodium glucoheptonate are mixed, stirred for reaction for 8-12 hours, and then allowed to stand for treatment, and filtered to obtain a high-concentration peracetic acid stock solution.

5. The method for simultaneous disinfection and sterilization of air and surfaces using a composite peracetic acid disinfectant according to claim 4, characterized in that The static treatment time is 4 to 6 days.

6. The method for simultaneous disinfection and sterilization of air and surfaces using a composite peracetic acid disinfectant according to claim 1, characterized in that In step 1, the concentration of peracetic acid in the high-concentration peracetic acid stock solution is 80-120 g / L, and the concentration of hydrogen peroxide is 40-70 g / L.

7. The method for simultaneous disinfection and sterilization of air and surfaces using a composite peracetic acid disinfectant according to claim 1, characterized in that In step 3, the amount of peracetic acid generated is controlled to be 10 mg / m 3 ~30 mg / m 3 The action time is 30min to 120min.

8. A method for simultaneous disinfection and sterilization of air and surfaces using a peracetic acid composite disinfectant, characterized in that The method for simultaneous disinfection and sterilization of air and surfaces using a peracetic acid compound disinfectant is implemented in the following steps:

1. Mix hydrogen peroxide, glacial acetic acid, 1-butyl-3-methylimidazole phosphotungstate, hydroxyethylidene diphosphonic acid and sodium glucoheptonic acid, stir and react for 8 to 12 hours, then let stand for treatment, and filter to obtain a high-concentration peracetic acid stock solution; 2. Use standard hard water to dilute the high-concentration peracetic acid stock solution into a peracetic acid solution with a concentration of 20g / L to 100g / L; 3. Load the peracetic acid solution into the hydrogen peroxide vaporization disinfector, control the flash temperature to 110℃~150℃, and blow the generated vaporized peracetic acid into the air or onto the surface of objects through a fan for disinfection and sterilization.

9. The method for simultaneous disinfection and sterilization of air and surfaces using a composite peracetic acid disinfectant according to claim 8, characterized in that In step 3, the generation rate of vaporized peracetic acid is controlled to be 0.8 mL / min to 1.2 mL / min, and the generation amount is 80 mg / m 3 ~120 mg / m 3 .

10. The method for simultaneous disinfection and sterilization of air and surfaces using a composite peracetic acid disinfectant according to claim 8, characterized in that The disinfection and sterilization time in step three is 30 minutes to 120 minutes.

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