Disinfection method

Abstract

The present disclosure relates to a method for disinfecting air and surfaces in a space. The method includes dispersing a disinfecting composition containing a diol (e.g., triethylene glycol) into the space at a frequency to maintain the disinfecting composition in the form of an aerosol at a concentration in the space ranging from about 0.02 mg / m3 to about 0.09 mg / m3.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 376,642, filed on September 22, 2022, and U.S. Provisional Application No. 63 / 481,725, filed on January 26, 2023, the contents of each of which are incorporated herein by reference in their entirety. Technical Field

[0003] The present disclosure relates to compositions and methods for disinfecting a space (eg, an indoor space). Background Art

[0004] Human diseases are usually caused by pathogenic microorganisms such as bacteria, viruses and fungi. The movement of infectious particles from the host or infected individuals to susceptible new victims can occur by various mechanisms, including breathing aerosolized fluids from the host, contact with surfaces contaminated by the host or host body fluids, or by spreading to the victim from the host or contaminated surfaces on the hands of the victim or a third party. Specific propagation mechanisms depend on organisms and specific environments. For example, in hospitals and other clinical settings, organisms such as Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter species (collectively referred to as ESKAPE pathogens) and Clostridium difficile can cause a large number of hospital-acquired infections. In addition, multidrug-resistant organisms (such as MRSA) (mainly bacteria) that are resistant to one or more classes of antimicrobial agents have special clinical significance due to their acquired resistance. Hospital-acquired infections have become a major problem in the healthcare industry. As another example, in the food preparation industry, large-scale food packaging facilities are periodically associated with outbreaks of antibiotic-resistant Salmonella enterica, resulting in numerous deaths. As another example, mold or fungus in a space may cause health-related problems and property damage. Therefore, there is a need for improved methods and compositions for disinfecting spaces in healthcare facilities, food packaging facilities, and other environments. Summary of the invention

[0005] The present inventors unexpectedly discovered that aerosols (e.g., droplets) containing triethylene glycol (TEG) can be maintained in the air of an indoor space at low concentrations while still effectively inactivating (e.g., killing) pathogens in the air and / or on surfaces in the indoor space, thereby effectively sanitizing (e.g., disinfecting or sterilizing) the indoor space (i.e., occupied or unoccupied by humans).

[0006] In one aspect, the disclosure features a method for disinfecting a space, the method comprising dispersing a disinfecting composition comprising triethylene glycol into the space at a frequency selected to maintain the disinfecting composition in aerosol form in the space at a range of about 0.02 mg / m 3 To about 0.09mg / m 3 concentration.

[0007] Other features, objects, and advantages will be apparent from the description and from the claims. DETAILED DESCRIPTION

[0008] As defined herein, unless otherwise indicated, all percentages expressed are to be understood as weight percentages relative to the total weight of the composition.

[0009] In general, the present disclosure relates to compositions and methods for disinfecting a space (e.g., an indoor space) by inactivating (or killing) pathogens (e.g., microorganisms or bacteria, viruses or fungi) in the space. Examples of suitable spaces include offices, schools, hotels, halls, theaters, reception rooms, bathrooms, health care facilities (e.g., nursing homes, wards (e.g., intensive care facilities) and medical offices (e.g., dental clinics)), food packaging facilities, agricultural farming facilities, indoor farms (e.g., poultry farms, pig farms or cattle farms), institutional kitchens, cafeterias, restaurants, public transportation (buses, trains, subways and airplanes), ambulances, indoor stadiums and sports facilities, law enforcement facilities (e.g., prisons), government facilities, elevators, retail outlets and other indoor public or private spaces. Typically, the disinfecting compositions described herein can be used to inactivate pathogens in unoccupied spaces (e.g., spaces not occupied by people) or occupied spaces (e.g., spaces occupied by people).

[0010] As used herein, the term "inactivating pathogens" means killing pathogens or otherwise inactivating pathogens. For example, the disinfecting compositions described herein can inactivate viruses, kill bacteria, kill mycobacteria, kill spores, kill mold and mildew.

[0011] In some embodiments, pathogens that can be inactivated by the disinfecting compositions described herein may include bacteria (e.g., gram-positive bacteria, gram-negative bacteria, or antibiotic-resistant bacteria), fungi (e.g., molds), viruses (e.g., enveloped viruses or non-enveloped viruses), and spores (e.g., spores produced by fungi or bacteria). Examples of pathogens that can be inactivated by the disinfecting compositions described herein include methicillin-resistant Staphylococcus aureus (MRSA; gram-positive bacteria), molds (e.g., Aspergillus species such as Aspergillus brasiliensis), Pseudomonas (e.g., Pseudomonas aeruginosa; gram-negative bacteria), Listeria (e.g., Listeria monocytogenes; gram-positive bacteria), Salmonella (e.g., Salmonella enterica; gram-negative bacteria), Klebsiella (e.g., Klebsiella pneumoniae; gram-negative bacteria), Mycobacteria (e.g., Mycobacterium tuberculosis), spores (e.g., anthrax spores, Clostridium difficile spores, or mold spores), or mixtures thereof. Examples of enveloped viruses that can be inactivated by the disinfecting compositions described herein include Lassa virus, Marburg virus, pneumonia, smallpox, croup virus, human parainfluenza virus (HPIV), respiratory syncytial virus (RSV), Ebola virus, German measles (rubella), herpes simplex (HSV), mumps, influenza (e.g., H1N1 virus), coronavirus (e.g., SARA-CoV-2 virus), and varicella. Examples of non-enveloped viruses that can be inactivated by the disinfecting compositions described herein include rhinoviruses, enteroviruses, and parvoviruses.

[0012] In some embodiments, the disclosure features a method of disinfecting a space (e.g., an indoor space). In some embodiments, the method may include dispersing (e.g., by dispersing, spraying, nebulizing, atomizing, or vaporizing) a disinfectant composition containing triethylene glycol into the space (e.g., an indoor space) to maintain the disinfectant composition in aerosol form in the space at a range of about 0.02 mg / m 3 To about 0.09mg / m 3 concentration, thereby inactivating (e.g., killing) any pathogens in the space (e.g., pathogens introduced into the space before or after the space is treated with the disinfecting composition). In some embodiments, dispersing the disinfecting composition can be performed by a system that generates an aerosol (visible or invisible to the human eye) in the space, such as a vaporizer (e.g., a smoke generator), a nebulizer (e.g., an odor dispersion unit), or an atomizer (e.g., a humidifier). When the aerosol is visible to the human eye, the aerosol can be in the form of fog, smoke, or haze. The system can be those known in the art, such as fog / smoke machines or smoke simulators used for emergency training or for the lighting industry to produce dramatic effects.

[0013] In some embodiments, the concentration of the disinfecting composition in aerosol form (e.g., the concentration of droplets containing the disinfecting composition) in the space may be at least about 0.02 mg / m 3 (e.g., at least about 0.03 mg / m 3 , at least about 0.04 mg / m 3 , at least about 0.05 mg / m 3 , at least about 0.06 mg / m 3 , at least about 0.07 mg / m 3 or at least about 0.08 mg / m 3 ) to a maximum of about 0.09 mg / m 3 (e.g., up to about 0.08 mg / m 3 , up to about 0.07mg / m 3 , up to about 0.06mg / m 3 , up to about 0.05mg / m 3 , up to about 0.04mg / m 3 or up to about 0.03 mg / m 3 For example, the concentration of the disinfectant composition in the form of an aerosol in the air may range from about 0.02 mg / m 3 To about 0.05mg / m 3 Typically, the above concentration range of the aerosolized disinfectant composition is significantly lower than the concentration range that can be detected by the human eye (i.e., at least about 0.5 mg / m 3 ), and therefore invisible to the human eye. Without wishing to be bound by theory, it was surprisingly found that the disinfectant composition in aerosol form was maintained at a concentration as low as about 0.02 mg / m 3 To about 0.09mg / m 3 The concentration of about 0.02 mg / m can effectively inactivate (or kill) various pathogens in the space (suspended in the air or on the surface). In addition, without wishing to be bound by theory, it is surprisingly found that although the concentration of the aerosolized disinfectant composition is maintained within the above concentration range (i.e., about 0.02 mg / m 3 To about 0.09mg / m 3 ) can effectively inactivate (or kill) various pathogens in the space, but increasing the concentration of the aerosolized disinfectant composition beyond this concentration range does not necessarily increase the efficacy of the composition. In addition, without wishing to be bound by theory, it is believed that the aerosolized disinfectant composition can generate a vapor containing a glycol (e.g., triethylene glycol or propylene glycol), which can inactivate pathogens in the space.

[0014] Typically, the aerosolized disinfectant composition can form a vapor containing TEG and other optional components in the disinfectant composition (e.g., another glycol, such as propylene glycol) in the treated space. In some embodiments, the total concentration of TEG (including TEG in the aerosol and TEG in the vapor) or glycol (for example, including both TEG and another glycol such as propylene glycol) in the space can be at least about 0.4 mg / m 3 (e.g., at least about 0.5 mg / m 3 , at least about 0.6 mg / m 3 , at least about 0.8 mg / m 3 , at least about 1 mg / m 3 , at least about 1.2 mg / m 3 , at least about 1.4 mg / m 3 , at least about 1.5 mg / m 3 , at least about 1.6 mg / m 3 or at least about 1.8 mg / m 3 ) or up to about 2 mg / m 3 (e.g., up to about 1.8 mg / m 3 , up to about 1.6 mg / m 3 , up to about 1.5 mg / m 3 , up to about 1.4 mg / m 3 , up to about 1.2 mg / m 3 , up to about 1 mg / m 3 , up to about 0.8 mg / m 3 , up to about 0.6mg / m 3 or up to about 0.5 mg / m 3 For example, the total concentration of TEG (including TEG in aerosol and TEG in vapor) or glycol (including glycol in aerosol and in vapor) in the air may be about 0.4 mg / m 3 About 0.7 mg / m 3 (e.g., about 0.5 mg / m 3 About 0.6 mg / m 3 ).

[0015] In some embodiments, in order to maintain the disinfectant composition in aerosol form at a desired level in a space (e.g., at about 0.02 mg / m 3 To about 0.09mg / m 3The methods described herein may include dispersing the disinfecting composition at a frequency that may depend on various factors, such as the size of the space, the temperature and humidity of the space, the airflow conditions in the space (e.g., the air exchange rate), the particle size of the aerosolized composition, and the amount of composition introduced into the space during each dispersion period. In some embodiments, the methods described herein may include dispersing the disinfecting composition for at least about 5 seconds (e.g., at least about 10 seconds, at least about 15 seconds, at least about 20 seconds, at least about 25 seconds, or at least about 30 seconds) to at most about 1000 seconds (e.g., at most about 900 seconds, at most about 800 seconds, at most about 700 seconds, at most about 600 seconds, at most about 500 seconds, or at most about 300 seconds) of a dispersion period (during which an amount of the disinfecting composition is introduced into the space), followed by a non-dispersion period (during which the disinfecting composition is not introduced into the space) of about 15 seconds (e.g., at least about 20 seconds, at least about 30 seconds, at least about 40 seconds, at least about 50 seconds, at least about 1 minute, at least about 2 minutes, at least about 4 minutes, or at least about 5 minutes) to about 10 minutes (e.g., at most about 8 minutes, at most about 6 minutes, at most about 5 minutes, at most about 4 minutes, at most about 2 minutes, or at most about 1 minute). In some embodiments, the methods described herein may include dispensing the disinfectant composition in an amount of at least about 2 grams (e.g., at least 3 grams, at least about 4 grams, or at least about 5 grams) to at most about 25 grams (e.g., at most about 20 grams, at most about 15 grams, at most about 10 grams, at most about 9 grams, at most about 8 grams, at most about 7 grams, at most about 6 grams, or at most about 5 grams) for a space having a volume of about 1000 cubic feet every 6 hours to maintain the aerosolized disinfectant composition at a desired concentration level (e.g., about 0.02 mg / m 3 To about 0.09mg / m 3 ).

[0016] In some embodiments, in order to maintain the disinfectant composition in aerosol form at a desired level in a space (e.g., at about 0.02 mg / m 3 To about 0.09mg / m 3The method described herein may include dispersing the disinfectant composition into the space as needed. In such embodiments, the method may include dispersing the disinfectant composition into the space by a system including an aerosol generator (e.g., a diffusion device) and a sensor. The aerosol generator may be configured to generate aerosol particles of an appropriate size (such as a particle size described herein) so that the aerosol particles can be suspended in the air. The sensor may be in fluid communication with the air in the space to be processed and configured to measure the concentration of the aerosolized disinfectant composition in the space. The measured concentration may be provided on a display on the sensor or provided to the user through a mobile application. In addition, the sensor may be configured to operate the aerosol generator to disperse the disinfectant composition into the space. The concentration of the aerosolized disinfectant composition in the space may be measured by a sensor, and the operation of the aerosol generator may be controlled to achieve the desired concentration of the aerosolized disinfectant composition in the space. The concentration measurement and dispersion steps may be repeated periodically as needed to maintain the desired concentration of the aerosolized disinfectant composition in the space. Examples of such systems are described in U.S. Provisional Application No. 63 / 365,729, the entire contents of which are incorporated herein by reference. In some embodiments, the above method can be automated so that the aerosol concentration can be automatically controlled within a desired range (e.g., about 0.02 mg / m 3 To about 0.09mg / m 3 ).

[0017] In some embodiments, the disinfectant composition can be in a stable state within the space to be treated (i.e., the concentration of the aerosolized disinfectant composition can be maintained at about 0.02 mg / m 3 To about 0.09mg / m 3 In the range of 5 to 24 hours per day) for a long period of time (e.g., 5 minutes to 24 hours per day). For example, the disinfecting composition can be maintained at a desired concentration in a steady state for at least about 5 minutes (e.g., at least about 10 minutes, at least about 20 minutes, at least about 30 minutes, at least about 1 hour, at least about 2 hours, at least about 3 hours, at least about 4 hours, or at least about 6 hours) per day to at most about 24 hours (e.g., at most about 18 hours, at most about 15 hours, at most about 12 hours, at most about 10 hours, or at most about 9 hours). Without wishing to be bound by theory, it is believed that the disinfecting composition described herein can be used in a relatively low aerosol concentration (e.g., about 0.02 mg / m 3 To about 0.09mg / m 3) forms saturated vapor, and the vapor pressure of saturated vapor can make aerosol particles form a uniform dispersion in the whole space in a stable state. Generally, vapor refers to a substance (e.g., a disinfecting composition) in the gas phase at a temperature below its critical temperature (e.g., by increasing the pressure on the vapor without reducing the temperature of the vapor, the vapor can be condensed into a liquid). In addition, generally, an aerosol refers to a suspension of droplets (e.g., droplets of a disinfecting composition) in the air or another gas. In addition, generally, saturated vapor refers to a substance (e.g., a disinfecting composition) in the gas phase, which will condense into a liquid phase under small changes in pressure and / or temperature.

[0018] In some embodiments, the aerosolized disinfectant composition can be maintained at a uniform concentration (i.e., at about 0.02 mg / m 3 To about 0.09mg / m 3 ) for a long period of time (e.g., 5 minutes to 24 hours).

[0019] In some embodiments, the method may further include vaporizing the disinfectant composition (e.g., in an aerosol generator such as a humidifier, a fog / haze machine, a smoke generator, a sprayer, or atomizer) before dispersing (e.g., spraying) the disinfectant composition into the space. In some embodiments, vaporizing the composition can be performed by treating the composition with steam or heat. For example, when vaporizing the composition by heating, the method may include delivering the composition to a heat exchanger to vaporize the composition. The heated steam can be forced through a nozzle as steam and / or droplets (or liquid particles) to form a visible or invisible aerosol. For example, when a vaporizer is used to disperse (e.g., spray) the disinfectant composition described herein into the space, the vaporizer can have a liquid reservoir and a pump (e.g., an electric pump) can be used to push the disinfectant composition in the liquid reservoir into a heat exchanger, where the disinfectant composition is vaporized. The heated steam is forced through a nozzle as steam and as droplets (or liquid particles) to form a visible or invisible aerosol.

[0020] In some embodiments, when a nebulizer (e.g., sprayer) is used to disperse (e.g., spray) the disinfection composition into a space, the composition can be converted into an aerosol by pressure. For example, the composition can be pumped or siphoned into a series of chambers of a specific size in a nebulizer or sprayer to increase the pressure and speed of the composition, thereby converting it from a liquid to an aerosol. In some embodiments, ultrasound and / or vibrating mesh technology can be used to disperse the composition into a space by a nebulizer or sprayer.

[0021] In some embodiments, the disinfection composition is dispersed to form vapor and / or droplets (or liquid particles) containing triethylene glycol (or other glycol). In some embodiments, the droplets can form an aerosol containing triethylene glycol (or other glycol). In some embodiments, the composition is dispersed to form an aerosol, vapor, or a mixture thereof. In some embodiments, the aerosol droplets may have an average diameter of at least about 10 nm (e.g., at least about 20 nm, at least about 50 nm, at least about 100 nm, at least about 200 nm, at least about 500 nm, at least about 1 μm, at least about 2 μm, or at least about 5 μm) to at most about 10 μm (e.g., at most about 8 μm, at most about 6 μm, at most about 5 μm, at most about 4 μm, at most about 2 μm, at most about 1 μm, or at most about 0.5 μm). In some embodiments, the methods described herein may be used per cm 3 A space (e.g., an indoor space) produces at least about 2000 (e.g., at least about 3000, at least about 4000, at least about 5000, at least about 6000, at least about 8000, or at least about 10,000) to at most about 150,000 (e.g., at most about 50,000 or at most about 25,000) droplets.

[0022] In some embodiments, the disinfection composition is dispersed (e.g., sprayed) can be intermittently (e.g., at constant intervals or at irregular intervals). In some embodiments, when the composition is applied intermittently at constant intervals, the frequency of application can be changed as needed depending on factors such as the concentration of triethylene glycol or other glycols in the composition, the temperature and humidity of the space, the size of the space, the desired concentration of the composition in the space, and the air exchange rate. In some embodiments, the preferred temperature of the space can be in the range of about 5°C to about 50°C (e.g., about 10°C to about 30°C or about 15°C to about 30°C). In some embodiments, the preferred relative humidity of the space can be in the range of about 5% to about 75% (e.g., about 15% to about 70%, about 30% to about 65%, or about 45% to about 60%). In some embodiments, the time period between two applications of the composition can be at least about 10 seconds (e.g., at least about 30 seconds, at least about 1 minute, at least about 1.5 minutes, at least about 2 minutes, at least about 5 minutes, at least about 10 minutes, at least about 30 minutes, or at least about 1 hour) to at most about 2 hours (e.g., at most about 1 hour, at most about 30 minutes, at most about 10 minutes, or at most about 5 minutes).

[0023] In some embodiments, the disinfecting composition can be dispersed (e.g., sprayed) continuously. In some embodiments, the disinfecting composition can be continuously dispersed into the space for at least about 5 minutes (e.g., at least about 10 minutes, at least about 30 minutes, at least about 1 hour, or at least about 2 hours) to at most about 24 hours (e.g., at most about 12 hours, at most about 9 hours, at most about 6 hours, at most about 5 hours, at most about 4 hours, at most about 3 hours, at most about 2 hours, or at most about 1 hour).

[0024] Without wishing to be bound by theory, it is believed that the disinfection composition or TEG at a concentration within the range described herein can effectively kill or inactivate at least 95% (e.g., at least about 96%, at least about 97%, at least about 98%, at least 98.5%, at least 99%, at least 99.5%, or at least 99.9%) of pathogens in a space within a short period of time (e.g., at most 60 minutes, at most 30 minutes, at most 15 minutes, at most 10 minutes, at most 5 minutes, at most 3 minutes, at most 2 minutes, at most 1 minute, or at most 30 seconds). In some embodiments, for certain pathogens (e.g., mold) that are known to be difficult to inactivate, the disinfection composition or TEG at a concentration within the range described herein may take a relatively long time (e.g., at least about 4 hours, at least about 8 hours, at least about 12 hours, or at least about 24 hours) to achieve the above-mentioned inactivation efficacy (e.g., at least about 95%). In some embodiments, at least about 0.5 grams (e.g., at least about 1 gram, at least about 2 grams, at least about 3 grams, or at least about 4 grams) and / or at most about 10 grams (e.g., at most about 5 grams) of the disinfecting composition can be applied to a space with a volume of 1000 cubic feet every 4 hours (e.g., every two hours or every hour).

[0025] In some embodiments, in order to practice the disinfection method described herein in an indoor space, the system described herein (e.g., a vaporizer or an atomizer such as a sprayer) can be placed in the center or one or more sides of the indoor space to be treated. In some embodiments, the system described herein can be incorporated into an HVAC system that controls the temperature, humidity and / or purity of the air in the indoor space. In such an embodiment, the system described herein may not need to be placed in the indoor space to be treated. In some embodiments, multiple systems can be used in appropriate locations to ensure uniform distribution of the disinfection composition. The disinfection composition described herein can be applied from the system to the indoor space until the desired disinfection (or sterilization) level is reached. In some embodiments, the disinfection composition can be applied continuously or intermittently (e.g., every minute or every 10 minutes) to maintain the desired disinfection level.

[0026] In some embodiments, the disinfecting composition described herein can be applied to the indoor space to be treated via an HVAC unit. For example, a system containing the disinfecting composition described herein can be connected to the return air plenum of the HVAC unit by a pipeline. The composition can then be applied to the indoor space by the HVAC unit until the desired disinfection (or sterilization) level is reached. This method can disinfect both the filter in the HVAC unit and the indoor space. In at least some embodiments, the composition will not be substantially removed from the air by the HVAC unit. For example, in some embodiments, at least some compositions in the form of droplets may be attached to the filter of the HVAC unit, but despite this, can still be vaporized and released from the filter and enter the environment.

[0027] In some embodiments, the space to be treated (e.g., indoor space) may include pathogens suspended in the air, and the disinfection method described herein can inactivate (e.g., kill) pathogens in the air. In some embodiments, the space may include pathogens on a surface (e.g., a hard or soft surface, or a non-porous or porous surface), and the disinfection method described herein can inactivate (e.g., kill) pathogens on the surface. In some embodiments, the surface can be any surface in the indoor space, such as the surface of a wall, floor, table, chair, computer, carpet, plant, or curtain. Without wishing to be bound by theory, it is believed that triethylene glycol (or other glycol) in the steam produced by the aerosolized disinfection composition can be attached to pathogens in the air or on the surface, to inactivate pathogens by denaturing or destroying the protein or membrane of the pathogen. In addition, without wishing to be bound by theory, it is believed that when the pathogen is a bacterium, mycobacterium, mold, or spore, the disinfection composition described herein can inactivate (e.g., kill) pathogens by drying.

[0028] In some embodiments, the disinfection composition described herein may include (e.g., contain, consist essentially of, or consist of) triethylene glycol (or other glycol) and water (e.g., deionized water). Triethylene glycol is miscible with water, has a boiling point of 286.5°C at a pressure of 101.325 kPa, and has a relatively low vapor pressure compared to water. Without wishing to be bound by theory, it is believed that the glycol (e.g., triethylene glycol or propylene glycol) in the vapor produced by the aerosolized disinfection composition inactivates the pathogen by condensing on the pathogen until the concentration of the glycol becomes high enough to denature the pathogen. In addition, without wishing to be bound by theory, it is believed that the glycol (e.g., triethylene glycol or propylene glycol) is highly hygroscopic and can inactivate (e.g., kill) the pathogen by absorbing water from the pathogen. In addition, without wishing to be bound by theory, it is believed that triethylene glycol (or propylene glycol) has very low acute or chronic toxicity when inhaled or ingested (especially at levels used in air for disinfecting (e.g., sterilizing) indoor spaces), and is therefore safe for use in indoor spaces (occupied or unoccupied).

[0029] Generally, the amount of glycol (e.g., triethylene glycol or propylene glycol) in the disinfection composition described herein is not particularly limited and can be varied as desired. For example, a disinfection composition containing a relatively low amount of triethylene glycol can achieve the same sterilization effect as a disinfection composition containing a relatively high amount of triethylene glycol by applying the former composition in an indoor space at a higher frequency or in a higher amount. In some embodiments, the disinfecting compositions described herein may include triethylene glycol in an amount of at least about 1% (e.g., at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 52%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, or at least about 80%) by weight of the composition to at most about 99.5% (e.g., at most about 99%, at most about 95%, at most about 90%, at most about 85%, at most about 80%, at most about 75%, at most about 70%, at most about 65%, at most about 60%, or at most about 50%) by weight. In some embodiments, triethylene glycol may be 100% (i.e., without any other ingredients) of the disinfecting compositions described herein. It is believed that application of a disinfecting composition containing relatively high amounts (eg, at least about 50% by weight) of triethylene glycol can increase germicidal efficiency and reduce the frequency of application of the composition.

[0030] In some embodiments, the water in the disinfection composition described herein is deionized water, reverse osmosis (RO) water, or ultrapure water (e.g., when used in a vaporizer). In some embodiments, the water may have a resistivity of at least 17 megohms, a total organic carbon content of at most about 10 ppb, and a bacterial count of at most about 10 CFU / ml. For example, the water may contain ions in an amount of at most about 50 ppm (e.g., at most about 40 ppm, at most about 30 ppm, at most about 20 ppm, at most about 10 ppm, at most about 5 ppm, or at most about 1 ppm) to at least about 1 ppb (e.g., at least about 10 ppb) of the total amount of water. In some embodiments, when the disinfection composition described herein is used in conjunction with a nebulizer (e.g., a sprayer), the water in the composition may be tap water (i.e., not deionized water, RO water, or ultrapure water).

[0031] In some embodiments, the disinfecting compositions described herein may include water in an amount of at least about 1% (e.g., at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 48%, at least about 50%, at least about 60%, or at least about 70%) by weight of the composition to at most about 99% (e.g., at most about 95%, at most about 90%, at most about 85%, at most about 80%, at most about 75%, at most about 70%, at most about 65%, at most about 60%, at most about 50%, or at most about 48%) by weight. Without wishing to be bound by theory, it is believed that the use of deionized water, RO water, or ultrapure water can minimize nozzle clogging (e.g., caused by the deposition of minerals in the water) of a system (e.g., a vaporizer) for applying the disinfecting compositions described herein, and thus can keep the system running for a long period of time. Furthermore, without wishing to be bound by theory, it is believed that the water in the disinfecting compositions described herein may facilitate the inactivation of pathogens by facilitating the evaporation of the glycol component of the disinfecting compound.

[0032] Without wishing to be bound by theory, it is believed that including water in the disinfectant composition can allow the composition to be easily sprayed, atomized or vaporized (e.g., by lowering the vaporization temperature and / or increasing the evaporation rate when the disinfectant composition is applied through an atomizer such as a sprayer, humidifier, fog / smoke machine or fog generator) and form an aerosol or vapor in the atmosphere. The water in the aerosol can evaporate quickly to form fine glycol droplets and / or vapor, which have a disinfecting effect and inactivate pathogens in the air or on surfaces. In addition, the water in the disinfectant composition can make the composition flammable, thereby producing a safer product than TEG alone (which is a flammable liquid with a flash point of 157°C).

[0033] In some embodiments, the disinfecting compositions described herein may further include optional ingredients, such as a glycol other than triethylene glycol. In some embodiments, the additional glycol may be propylene glycol. Without wishing to be bound by theory, it is believed that the additional glycol may increase the disinfecting effect of the composition. In some embodiments, the disinfecting compositions described herein do not include any additional glycol or any components other than triethylene glycol and water.

[0034] In some embodiments, the disinfecting compositions described herein may include an additional glycol (e.g., propylene glycol (PG)) in an amount of at least about 0.5% by weight (e.g., at least about 1%, at least about 1.5%, at least about 2%, at least about 2.5%, at least about 3%, at least about 3.5%, at least about 4%, at least about 4.5%) to at most about 99% by weight (e.g., at most about 95%, at most about 90%, at most about 80%, at most about 70%, at most about 60%, at most about 50%, at most about 40%, at most about 30%, at most about 20%, at most about 10%, at most about 5%, at most about 4.5%, at most about 4%, at most about 3.5%, at most about 3%, at most about 2.5%, at most about 2%, at most about 1.5%, or at most about 1%) of the composition.

[0035] In some embodiments, the disinfecting compositions described herein may optionally include substances that are generally recognized as safe ("GRAS") as defined by the U.S. Food and Drug Administration. Examples of suitable GRAS substances include dimethyl ether, glycerol, chlorine dioxide, and hypochlorous acid.

[0036] In some embodiments, the disinfecting compositions described herein may include (e.g., contain, consist essentially of, or consist of) about 50% to about 90% by weight of triethylene glycol and about 10% to about 50% by weight of water. In some embodiments, the disinfecting compositions described herein may include (1) triethylene glycol in an amount of about 52% to about 90% by weight of the composition; (2) water in an amount of about 5% to about 48% by weight of the composition; and (3) propylene glycol in an amount of about 0% to about 5% by weight (e.g., about 0.5% to about 5% by weight) of the composition. In some embodiments, the disinfecting compositions described herein may include about 52.25% by weight of triethylene glycol, about 1% by weight of propylene glycol, and about 46.75% by weight of deionized water.

[0037] The following examples are illustrative and are not intended to be limiting.

[0038] Example 1: Evaluation of the efficacy of disinfectant compositions against MS2 bacteriophage in aerosols

[0039] The efficacy of disinfection composition #1 against MS2 bacteriophage was tested at various airborne concentrations using two different test devices, namely, Hurricane 1800Flex (Chauvet DJ, Sunrise, FL) and Amhaze Stadium (Chauvet DJ, Sunrise, FL). Disinfection composition #1 contained triethylene glycol (52.25 wt%), propylene glycol (1 wt%), and deionized water (46.75%).

[0040] The MS2 bacteriophage (MS2) ATCC15597-B1 was selected for this test. The virus is a non-enveloped positive-strand RNA virus of the phage family Leviviridae. Bacterial cells are hosts for the phages, and for the MS2 phage, Escherichia coli 15597 was used for this purpose. Its small size, icosahedral structure, and environmental resistance make MS2 ideal for use as a surrogate virus.

[0041] MS2 is grown on an appropriate medium. Cultures used for test inocula are evaluated for sterility, harvested, washed in sterile phosphate-buffered saline and concentrated. Virus concentrations are determined after incubation at 36 ± 1 °C for 18-24 hours. MS2 samples are counted in 50% tryptic soy agar using standard dilution and plating techniques. The test inoculum is divided into two equal parts and added to the appropriate number of sprayers. No more than 20 ml of liquid culture should be added to each sprayer. The test parameters are as follows:

[0042] Inoculum volume added to nebulizer: 20 mL

[0043] Sampler medium (volume): Phosphate buffered saline with 0.1% Tween 80 (20 ml)

[0044] Sampling time: 10 minutes

[0045] Sampling type: Impinger, SKC biological sampler

[0046] Incubation temperature: 36±1℃

[0047] Virus atomization time: 60 minutes

[0048] Neck rinse medium (volume): Phosphate buffered saline (5 mL)

[0049] Disinfectant composition contact time: 0, 3 minutes, 15 minutes and 27 minutes

[0050] Counting medium: 50% tryptic soy agar

[0051] Incubation time: 18-24 hours

[0052] Test substances:

[0053] Disinfectant composition #1: triethylene glycol (52.25 wt%), propylene glycol (1 wt%), and DI water (46.75%)

[0054] Number of samples: 3

[0055] The test device is placed on the floor of the test room in a completely horizontal position at a distance of 5 feet from the low level carrier position. The test device is plugged into the socket and then the wired timer controller is plugged into the remote connector socket on the test device. The test device is allowed to heat up for 3 to 5 minutes. With the fluid intake tube in the test substance bottle, the manual button on the remote control is pressed to prime the machine. Then, the test disinfecting composition is weighed after priming and placed back into the test device for testing.

[0056] Before the test begins, set up the test room and complete the safety checklist. The test is started by aerosolizing MS2 for 60 minutes using a nebulizer and bringing the concentration to the desired amount. Once the target virus concentration is reached, a zero-time sample is collected, and then the test device is run for the specified contact time, and additional samples are collected for each contact time. In the baseline run, no test composition is added to the test chamber, and samples are collected at 3 minutes, 15 minutes, and 27 minutes to determine the death and sedimentation of the virus at these times. In the test run, the test device is run for a predetermined time (i.e., 30 seconds for the Amhaze Stadium device and 3 seconds for the Hurricane 1800Flex device) to add the target amount of disinfectant composition #1 to the test chamber, and samples are collected at each contact time (3 minutes, 15 minutes, and 27 minutes). By using SKC All samples were taken for 10 minutes in phosphate buffered saline (20 mL) with 0.1% Tween 80. The amount of disinfecting composition #1 used by the Amhaze Stadium device and the Hurricane 1800Flex device was 51 grams and 13 grams, respectively.

[0057] Once testing was complete, a decontamination process was performed using a 4-hour UV exposure before any scientist entered the testing room. The reduction in MS2 was calculated relative to the concentration of MS2 at time zero or the corresponding control run sample (if applicable).

[0058] Calculate the virus concentration in the test chamber using the following equation:

[0059] PFU / ml = (average plate count) × 1:10 serial dilution factor

[0060] PFU / m 3 =[(PFU / ml×Vs)÷(Ts×12.5L / min)]×(1000L / m 3 )

[0061] Where Vs = volume of biological sampler (ml), Ts = sampling time (min). In addition, the Log of virus concentration was calculated by the following equation: 10 Reduction and reduction percentage:

[0062] Reduction percentage = [(BA) / B] × 100%

[0063] Log 10 Reduction = Log(B / A)

[0064] Log compared to the baseline 10 Reduce = Test Log 10 Reduce-Baseline Log 10 reduce

[0065] Where B = the number of live test microorganisms at time zero after aerosolization; and A = the number of live test microorganisms after the contact time.

[0066] The test results are summarized in Table 1 below.

[0067] Table 1

[0068]

[0069] As shown in Table 1, the test involving the Amhaze Stadium device (which used 51 grams of disinfecting composition #1) showed a net reduction percentage of 98.44%, 98.92%, and at least 98.71% at 3 minutes, 15 minutes, and 27 minutes, respectively. In addition, the test involving the Hurricane 1800Flex device (which used 13 grams of disinfecting composition #1) achieved a net reduction percentage of 99.76%, at least 99.92%, and at least 99.89% at 3 minutes, 15 minutes, and 27 minutes, respectively.

[0070] Example 2: Evaluation of the efficacy of disinfectant compositions against MS2 bacteriophage in aerosols at reduced amounts

[0071] Disinfecting composition #1 was tested for efficacy against MS2 bacteriophage at reduced amounts using the GLP protocol with Amhaze Stadium (Chauvet DJ, Sunrise, FL).

[0072] The test procedure is described below.

[0073] Handling glassware

[0074] Rinse the specialized glassware required for aerosol testing, including the nebulizer and SKC Biosampler, with tap water, then rinse with deionized water. Carefully wrap the glassware in microfiber or other soft material, place in a sterilization bag, and autoclave. Allow the glassware to dry at room temperature or at elevated temperatures. Record the drying conditions at the time of the study. Allow the glassware to equilibrate to room temperature before use in the study.

[0075] Aerosol chamber setup

[0076] Clean the aerosol test chamber (including walls, glove ports, and sampling locations) with a 1:100 dilution of household bleach. Record the disinfectant solution at the time of the study. Clean within 5 days of the start of testing and between test runs. After cleaning, wipe surfaces with sterile deionized water and allow surfaces to dry completely. On each day of testing and before each run, wipe the biosampler port with a 1:100 dilution of household bleach sprayed on a microfiber cloth. Record the disinfectant solution at the time of the study. Wipe the port and grommet thoroughly with the disinfectant solution and allow to dry completely.

[0077] Inoculum preparation

[0078] For MS2 bacteriophage ATCC 15597-B1: Store the prepared virus stock at -70°C ± 10°C until ready for testing. Remove the frozen stock from the freezer on the day of testing and allow to thaw. Grow host cultures in 10 mL of tryptic soy broth at 36 ± 1°C for 6-24 hours. Prepare the stock by diluting to ≥ 1.0 × 10 7 The inoculum was prepared at a target concentration of PFU / ml. The inoculum was counted. The nebulizer was prepared by adding 15-20 mL of the prepared inoculum inside the biosafety cabinet and then replacing the cap on the nebulizer before transferring to the chamber.

[0079] Aerosol efficacy room operation

[0080] Four chamber runs were performed. One was a baseline run, which was used to determine the natural settling and death of the test microorganisms after aerosolization. Three test runs were performed, in which the chamber was treated with each test substance batch using the test device.

[0081] Complete the safety checklist, including placing backup power, verifying HEPA filter functionality, checking UV lamps for proper operation, covering outlets, starting pressurized air, verifying sampler pumps, and checking chamber pressure. When the safety and pre-study checklists are completed to the point of adding the nebulizer, an appropriate N95 mask is donned by the participating analyst. The nebulizer is then placed and connected to pressurized air. The removable UV bulb is placed into the room, the chamber is exited, and the door is closed and sealed. Once nebulization begins, the door will not be reopened until decontamination occurs.

[0082] Before starting atomization, record the temperature and humidity. Start pressurized air and confirm that the nebulizers are running by visually observing that all six jets in both nebulizers are running. Allow the nebulizers to run for 60 minutes ± 30 seconds. Once atomization is complete, record the temperature and humidity.

[0083] Weigh and record the amount of Disinfection Composition #1 loaded into the apparatus. Operate the test apparatus until Disinfection Composition #1 is observed to flow from the apparatus. Prepare the apparatus outside the chamber for ≥5 minutes. Record the weight of Disinfection Composition #1 used during the preparation period. The test apparatus is then set up and operated by the operator. The operator records any data or apparatus settings related to the dosage of Disinfection Composition #1 entering the chamber. During the entire test run, Disinfection Composition #1 is added to the test chamber to achieve and maintain the preset aerosol concentration. Record the weight of Disinfection Composition #1 used during the test.

[0084] Aerosol collection

[0085] Use biological sampler, collect sample (including zero time) after atomization and subsequently after predetermined exposure time at intervals. Make sampler run for 10 minutes ± 10 seconds. According to manufacturer's suggestion, vacuum pump maintains 10-20psi pressure, so that sampler runs with 12.5L / min. After treatment time begins, collect sample in triplicate at following sampling time: zero time, 30 seconds, 15 minutes, 60 minutes. During sampling time, verify that sampler is sampling properly by observing the eddy current produced in sampler.

[0086] Handling Biological Samplers

[0087] Once the sample has been collected, move the sampler to a biosafety cabinet for processing. Rinse each neck with 5 mL of sterile phosphate buffered saline. Allow the liquid to drain into the collection cup of the container. Transfer the liquid to a sterile 50 mL conical container or equivalent sterile container. During the transfer, observe and record the total liquid volume. Then count the sample. All cultures and collected samples are counted by ml. As needed, the sample is diluted in phosphate buffered saline with a 1:10 dilution to observe the countable range of plaques. For this assay, the countable range is considered to be 25-250 colonies / plate. For a dilution series with only counts <25 on the least diluted plate, counts less than 25 can be used in the calculation. For a dilution series without counts, the detection limit is calculated, and this can be used to estimate the log reduction. It is preferred to use counts within 25-250, however, counts outside this range can also be used. For counts>250, it is recorded as too many to count (TNTC). The dilution is plated using the pour plate technique.

[0088] MS2 phage ATCC 15597-B1 was plated onto 50% TSA supplemented with 0.100 ml per plate of E. coli ATCC 15597. The plates were swirled well to mix and then allowed to solidify before incubation.

[0089] Aerosol Chamber Purification

[0090] Start all UV lamps equipped in the aerosol chamber to facilitate decontamination and allow them to run for at least 2 hours. Disconnect the air compressor and store it appropriately.

[0091] Neutralization Verification Laboratory Operation

[0092] To evaluate the neutralization of the test substances, a neutralization validation chamber run was performed on a representative batch of disinfectant composition #1. Neutralization of each combination of test microorganisms and disinfectant composition #1 was evaluated. A set of safety checks was performed and recorded at the time of the test. Once the safety and pre-study checklists were completed to the extent of adding the sprayer, the appropriate N95 mask was worn by the participating analysts. A removable UV bulb was placed in the room. The amount of disinfectant composition #1 loaded into the device was weighed and recorded. The test device was prepared until the disinfectant composition #1 was observed to flow out of the device. The device was prepared outside the room for ≥5 minutes. The chamber was treated with disinfectant composition #1. To simulate the test conditions, disinfectant composition #1 was released into the chamber by using the test device. Using an SKC biosampler filled with 20mL of neutralizer (PBS w / 0.001% Tween 80), the pump was turned on and air samples were collected for 10 minutes ± 10 seconds. According to the manufacturer's recommendations, the vacuum pump maintained a pressure of 10-20psi, allowing the sampler to operate at 12.5L / min. During the sampling time, verify that the sampler is sampling properly by observing the vortex generated within the sampler.The SKC Biosampler is then transferred to a biosafety cabinet and handled as described in the Handling Biosamplers section above.

[0093] Three analyses were prepared to ensure that equivalent levels of microorganisms were recovered from an inert control suspension (PBS), a suspension of a neutralizer alone, and a suspension of neutralized disinfectant composition #1 recovered from air as described above. Each of the above tubes was evaluated in equal volumes, where they were inoculated with 10-100 PFU of the test microorganism, vortexed, and then 1.0 ml aliquots were plated in duplicate. Multiple tubes of the three liquid types (PBS, neutralizer, and neutralized disinfectant composition #1) were inoculated to ensure that the target concentration of microorganisms was met. Multiple dilutions were performed and all data were recorded and included in the final report.

[0094] Counting of air samples

[0095] Incubate the counting plates of MS2 phage ATCC 15597-B1 and controls at 36°C ± 1°C for 18-24 hours.

[0096] Sterility control

[0097] On each day of use, a 1.0 ml aliquot of the neutralizer was plated. On each day of use, a 1.0 ml aliquot of phosphate buffered saline was plated. Plates containing only growth medium were incubated while testing. Purity streaks of the host microorganism were performed on each day of testing. Infectivity controls were performed where dilutions of the test inoculum were plated with the host microorganism to confirm infectivity. All sterility controls were incubated with the test plates.

[0098] The test results were calculated using the same equation outlined in Example 1 and are summarized in Table 2 below.

[0099] Table 2

[0100]

[0101] In this example, disinfecting composition #1 was intermittently added to the test chamber to achieve and maintain 0.437 mg / m 3 The test was performed in triplicate, and the average weight of disinfection composition #1 used during each test was 2.75 grams. As shown in Table 2 above, an average reduction percentage of 99.53%, 99.80% and 99.98% was achieved at 30 seconds, 15 minutes and 60 minutes, respectively, compared to time zero.

[0102] Example 3: Evaluation of the efficacy of disinfectant compositions against MS2 bacteriophage in aerosols at reduced amounts

[0103] Disinfecting composition #1 was tested for efficacy against MS2 bacteriophage at reduced amounts using a controlled release method using an Aura sprayer (Air Essentials, Miami, FL). The testing conditions and parameters are summarized below.

[0104] Test Material: Disinfectant Composition #1.

[0105] Test substance aerosol concentration: about 0.04mg / m 3 .

[0106] Test organism: MS2 bacteriophage.

[0107] Atomization stock solution concentration: 7.0x10 10 pfu / ml.

[0108] Atomization time: 20 minutes.

[0109] Room size: 16m 3 .

[0110] Exposure temperature: 22-24℃

[0111] Exposure humidity: 30-40% relative humidity.

[0112] Exposure time: Exposure time for this study was assessed as the amount of time of aerosol exposure.

[0113] Aerosol sampling duration: 10 minutes.

[0114] Aerosol sampling time points: 0 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, and 90 minutes.

[0115] Sedimentation slide duration: 90 minutes.

[0116] Neutralizer: Phosphate buffered saline solution + 0.05% Tween 80.

[0117] Agar plate medium: Tryptic soy agar.

[0118] Test Method

[0119] The test procedure is described below.

[0120] Chamber Preparation

[0121] Before the start of the test and between each test, wipe the chamber clean with 95% isopropyl alcohol and a non-fibrous towel. Clean all ports with a non-fibrous brush. Allow all surfaces to dry before the start of the test. For each cleaning of the chamber, record the lot number of the cleaning solution and the time and date of the cleaning.

[0122] Preparation of test organisms

[0123] Take 10 mL of previously prepared stock solution of MS2 bacteriophage (ATCC #15597-B1) from a -80°C freezer and thaw. The characteristics of the frozen stock used in this study were >1.0x10 11 pfu / ml. After thawing, the frozen stock was diluted 1:4 with phosphate buffered saline solution for testing. 50 ml of this prepared test inoculum was placed in the nebulizer cup of a Collison 24-jet nebulizer for testing. 5 mL of host culture E. coli ATCC #15597 was prepared in a 15 mL conical tube in tryptic soy broth and grown overnight in an incubator at 35 ± 2°C. After sampling, this host bacteria was used for plating of phage.

[0124] Neutralization Verification

[0125] Neutralization verification tests were run to ensure that all kill assessed by the test unit was due to kill in the air and not in the neutralizer PBS solution after sampling and during plating.

[0126] For the neutralization verification test, the amount of disinfectant composition #1 was based on a predetermined concentration of 0.04 mg / m 3To verify that any fluctuations in this concentration would not cause problems, validation was also performed at concentrations 1 log (10%) and 2 log (100%) higher than the concentration estimated in the chamber. Disinfection Composition #1 at these concentrations was diluted in PBS + 0.005% Tween 80. After thawing the MS2 phage from a frozen stock, it was diluted to less than 1.00 × 10 3 pfu / mL and added to each conical tube. The contact time for testing in the PBS mixture was 15 minutes. After 15 minutes, all samples were diluted, plated, incubated and counted for concentration. The concentration of each sample was compared with the control with only PBS solution and the control with samples diluted in sterile deionized water to confirm neutralization.

[0127] Chamber Test

[0128] Bioaerosol testing of disinfection composition #1 against MS2 bacteriophage consisted of a single control and a single test. The control was run to account for any natural mortality of this indoor airborne organism over time and to establish an untreated baseline concentration to compare with the test concentrations.

[0129] Along with the bioaerosol aspect of the test, a blank slide was placed in the chamber to assess the amount of fallout produced by the microorganisms. A 1" x 2" slide was placed on a table located 4 feet from the Aura dispersion device. The Aura reservoir was filled with Disinfecting Composition #1 in preparation for the test. The Aura was adjusted to the settings determined in the Concentration Matching section of the protocol. The device was then primed before being placed in the chamber. The prime consisted of running the device outside of the chamber for 20 minutes. This 20 minute prime ensured that the lines of the device were filled with fresh Disinfecting Composition #1 and that the device was operating correctly.

[0130] Before introducing bioaerosol, the device is placed on a stainless steel table, in the center of the room, 3 feet from the floor of the room. Before closing the door of the room, check the device to ensure that the appropriate settings are still enabled. After the door of the room is locked, check all outlets of the room to ensure that they are sealed, check all vacuum pumps to ensure that they are working, and verify that the pressurized air is working. After the inspection is completed, the device is remotely opened, and disinfection composition #1 is dispersed in the air of the room for 30 minutes to maintain the preset aerosol concentration before introducing the test bioaerosol. These 30 minutes are used as the pre-conditioning of the room, to simulate the environment that has been treated with disinfection composition #1. Before testing, the atomized stock solution is atomized at 40psi for 20 minutes. After the bioaerosol is atomized in the air of the room, the room is kept sealed until the purification and emptying of the room are completed. Temperature and humidity are recorded during each test.

[0131] Aerosol collection

[0132] All samples were collected using an AGI-30 impactor filled with 20 mL of sterile PBS with 0.005% Tween 80 added. The addition of Tween 80 was used to increase the impactor collection efficiency and for de-agglomeration of all microorganisms. The impactor simultaneously sampled from opposite corners of the chamber. After sampling, the impactors were pooled and collected in conical tubes for plating. The impactor sampled the chamber at 12.5 l / min using suction from a vacuum pump. Before the study began, each impactor used in the study was flow characterized. Aerosol samples were collected at 0, 15, 30, 45 and 60 minutes. All samples were collected for 10 minutes.

[0133] Settling plate collection

[0134] After the test is completed and the chamber is emptied, the sedimentation plate is removed from the chamber. Each plate is placed in a separate, individually labeled 50 mL conical tube containing 10 mL of sterile phosphate buffered saline solution + Tween 80 to be plated for concentration. Prior to plating, each conical tube is vortexed for 2 minutes and then vigorously shaken for another minute. The samples are then plated for concentration determination using the techniques described in the sample plating section below.

[0135] Sampler cleaning

[0136] Between each sample, the impactors were immersed in a 10% bleach solution and then immersed in DI water. They were then washed in a sink with high pressure tap water for a total of six times, followed by a final DI water rinse. The impactors were then air dried before being used for another sample.

[0137] Sample plating

[0138] For sample plating, overnight host bacteria (Escherichia coli ATCC #15597) were diluted 1:9 in PBS. 1.5 mL microcentrifuge tubes were filled with 800 μl sterile PBS and 100 μl of diluted host bacteria stock solution from the incubator. The number of tubes required is based on the number of samples to be plated and the predicted dilution range required for sample plating. Using this set of dilution tubes, the test samples were serially diluted. For this study, a standard droplet plaque assay technique was established for plating. The countable range of this assay is 5-50 plaques. Counts>50 pfu are marked as too many to count (TNTC). The samples were plated in triplicate on pre-labeled TSA culture plates with their respective dilution ranges. The plates were dried and then placed in an incubator set at 35 ± 2 ° C for 24 hours.

[0139] Incubation and observation

[0140] The plates were incubated at 35 ± 2°C for 20-24 hours before being removed for counting. Plate counts were recorded in a laboratory notebook at the time of counting and transferred to a Microsoft Excel spreadsheet for analysis.

[0141] Study Control

[0142] Control count

[0143] The MS2 phage aerosol inoculum produced >1.0×10 9 PFU / m 3 Acceptable starting concentration counts.

[0144] Sterility control

[0145] Sterility testing of the culture medium showed no growth of contaminants.

[0146] Culture purity

[0147] The host culture showed no signs of contamination when streaked on tryptic soy agar plates.

[0148] Neutralization control

[0149] The neutralizer produced no signs of contaminant growth after plating.

[0150] Data analysis

[0151] Calculation of % reduction

[0152] The starting bioaerosol concentration of the test run was compared to the starting concentration of the control run to calculate the effect of pre-treating the chamber by disinfection composition #1. The nebulization method and nebstock remained the same from the control run to the test run, so the starting bioaerosol concentration of the test run was expected to be the same as the control. Therefore, the changes in the starting bioaerosol concentration observed in the test run were attributed to the pre-treatment of the chamber.

[0153] To determine the log reduction at each time point in the experiment, the percentage of viable virus remaining airborne in the chamber was compared to the initial bioaerosol concentration in the uninhibited control experiment. This remaining percentage was then multiplied by the log 10 function to obtain the logarithmic reduction.

[0154] For the net reduction, the natural reduction from the control run was subtracted from the test run to account for the natural reduction not caused by Disinfecting Composition #1.

[0155] Compare the settlement plates from the control trial directly to the settlement plates from the test trial to assess the reduction. The difference between the control trial and the test trial is multiplied by the log 10 function to obtain the log reduction. Since natural mortality should be the same between the two trials, the reduction was assessed as a net reduction.

[0156] For a viable impact, the aerosol concentration is collected (C a ) is calculated as follows:

[0157] C a =(C imp ×I vol ×t) / Q imp

[0158] The effective aerosol concentration is collected (C a ) is cfu or pfu per liter of room air; effective impactor concentration collection (C Imp ) is the counted cfu or pfu / ml from the impactor sample; the impactor sample collection volume (I vol ) is 20 mL of collection fluid / impinge; the impactor sample flow rate (Q imp ) was 12.5 L / min; and the impactor sampling time (t) was 5 or 10 minutes, depending on the test.

[0159] The effective delivery efficiency of the aerosol system (expressed as %) is calculated as follows:

[0160] Efficiency = (C a / V p )×100%

[0161] Where V p is the effective particles per liter of air in the chamber. All test results were averaged to obtain the mean and standard deviation for each test set. The total effective bioagent concentration and the reduction over time were expressed as the log reduction of the aerosolized bioagent and were calculated for each test.

[0162] The test results were calculated using the same equation outlined in Example 1 and are summarized in Table 3 below.

[0163] Table 3

[0164]

[0165] As shown in Table 6, the disinfection composition #1 was maintained at a very low aerosol concentration in the air (about 0.04 mg / m 3) unexpectedly showed an average net Log after 15-25 min exposure to MS2 bacteriophage in aerosol 10 The pathogen reduction was 2.61. In other words, the composition was able to inactivate over 99% of MS2 bacteriophage in aerosols in only 15-25 minutes. The above results indicate that disinfection composition #1 will effectively inactivate MS2 bacteriophage in aerosol form in a space.

[0166] In addition, the sedimentation test showed that the average concentration of MS2 bacteriophages deposited on the slides was 3.02E+06 pfu / slide in the control experiment (where Disinfectant Composition #1 was not introduced into the test chamber) and 6.37E+02 pfu / slide in the test experiment (where Disinfectant Composition #1 was introduced into the test chamber). In other words, the results showed that Disinfectant Composition #1 was able to inactivate 99.98% (i.e., Log 1.0) of the MS2 bacteriophages in the air during 90 minutes. 10 The above results also show that disinfection composition #1 can effectively inactivate pathogenic MS2 bacteriophage in the air.

[0167] Other embodiments are within the following claims.

Claims

1. A method for disinfecting an indoor space, comprising: The disinfectant composition comprising triethylene glycol is dispersed into the space at a frequency selected to maintain the disinfectant composition in aerosol form in the space at a range of about 0.02 mg / m 3 To about 0.09mg / m 3 concentration.

2. The method of claim 1, wherein the method comprises dispensing the disinfecting composition for a dispensing period of about 5 seconds to about 1000 seconds, followed by a non-dispensing period of about 15 seconds to about 10 minutes.

3. The method according to claim 1 or 2, wherein the total amount of triethylene glycol in the space is maintained in the range of about 0.4 mg / m 3 About 2 mg / m 3 concentration.

4. The method according to any one of claims 1 to 3, wherein the concentration of the disinfecting composition in aerosol form in the space is in the range of about 0.02 mg / m 3 To about 0.05mg / m 3 .

5. The method according to any one of claims 1 to 4, wherein the disinfecting composition is in a stable state in the space.

6. The method according to any one of claims 1 to 5, wherein the method forms the disinfecting composition in the space that is invisible to the human eye.

7. The method of any one of claims 1-6, wherein the amount of triethylene glycol is from about 10% to about 90% by weight of the disinfecting composition.

8. The method according to any one of claims 1 to 7, wherein the disinfecting composition further comprises water.

9. The method of claim 8, wherein the amount of water is from about 5% to about 90% by weight of the disinfecting composition.

10. The method of any one of claims 1-9, wherein the disinfecting composition further comprises propylene glycol.

11. The method of claim 10, wherein the amount of propylene glycol is from about 0.5% to about 20% by weight of the disinfecting composition.

12. The method of any one of claims 1-11, wherein the disinfecting composition comprises about 10% to about 90% by weight of triethylene glycol and about 10% to about 90% by weight of water.

13. The method of any one of claims 1-12, wherein the disinfecting composition comprises about 52.25 weight percent triethylene glycol, about 1 weight percent propylene glycol, and about 46.75 weight percent water.

14. The method according to any one of claims 1 to 13, wherein the disinfecting composition is dispersed to form a mixture comprising an aerosol and a vapor.

15. The method of claim 14, wherein the aerosol comprises droplets having an average diameter of about 10 nm to about 10 μm.

16. The method according to any one of claims 1 to 15, wherein the method is performed by a nebulizer or a vaporizer.

17. The method of any one of claims 1-16, wherein the selected frequency is determined based on sensor data representing a concentration of the disinfecting composition in the space.

18. The method according to any one of claims 1 to 16, wherein: The selected frequency is a predetermined frequency.

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