Compounded formulation for air disinfection

Abstract

The present invention provides a disinfectant composition comprising 0.01-5 parts by weight of a compound of formula (I), 1-10 parts by weight of benzalkonium chloride, and the remainder being pure water. By compounding ethylicin and benzalkonium chloride, the present invention can effectively eliminate environmental pathogens at a very low dosage, thereby achieving a good disinfection effect.

Description

A compound preparation for air disinfection Technical Field

[0001] This invention relates to the field of environmental hygiene, and more specifically, to a compound preparation for air disinfection. Background Technology

[0002] Environmental disinfection refers to the elimination or inhibition of pathogens, viruses, bacteria, fungi, etc. in the environment to prevent the spread of diseases caused by pathogen contamination. With the increasing frequency of pathogen infections in recent years, the significance of environmental disinfection has become more prominent, and choosing an ideal disinfectant is the key to doing a good job in the prevention and control of infectious diseases.

[0003] Ethylene is a plant-based biomimetic pesticide with the chemical formula C4H. 10 O2S2 is a colorless or slightly yellow oily liquid with a garlic-like odor. It is an ethyl homologue of allicin, and its modified chemical structure is shown in the following formula:

[0004] Since the beginning of the 21st century, the compounding of chemical disinfectants has become an important research direction. Combinations of multiple disinfectant agents can synergistically enhance each other, broadening the antibacterial spectrum of disinfectants and improving their killing power against pathogenic microorganisms, resulting in a series of highly efficient and broad-spectrum disinfectants. Previous studies have shown that allicin is a highly efficient broad-spectrum biomimetic bactericide, but its strong garlic odor limits its use in indoor environmental disinfection. Therefore, there is a need in this field to develop a novel, odorless, easy-to-use, and highly efficient compound formulation for environmental disinfection.

[0005] Summary of the Invention

[0006] The purpose of this invention is to provide an odorless, non-toxic, and highly effective environmental disinfection compound preparation.

[0007] A first aspect of the present invention provides a disinfectant composition comprising:

[0008] Compound (I) 0.01–5 parts by weight,

[0009] 1-10 parts by weight of benzalkonium chloride; and

[0010] The remaining amount of pure water.

[0011] In another preferred embodiment, the disinfectant composition comprises: 0.1 to 3 parts by weight of compound of formula (I), 1 to 8 parts by weight of benzalkonium chloride, and the balance being pure water.

[0012] In another preferred embodiment, the disinfectant composition comprises: 0.5 to 2 parts by weight of compound of formula (I), 2 to 6 parts by weight of benzalkonium chloride, and the balance being pure water.

[0013] In another preferred embodiment, the disinfectant composition comprises the following components:

[0014] 0.05-0.2 wt% of compound of formula (I);

[0015] 0.1-1.0 wt% benzalkonium chloride; and

[0016] The remaining amount of pure water.

[0017] In another preferred embodiment, the disinfectant composition comprises 0.08-0.12 wt% of a compound of formula (I);

[0018] 0.4-0.5 wt% benzalkonium chloride; and

[0019] The remaining amount of pure water.

[0020] In another preferred embodiment, the disinfectant composition is an air disinfectant.

[0021] In another preferred embodiment, the disinfectant composition has a sterilization rate of ≥95%.

[0022] In another preferred embodiment, the LD50 of the disinfectant composition is... 50 >5000 mg / kg·bw.

[0023] In another preferred embodiment, the LC of the disinfectant composition 50 >10000mg / m 3 .

[0024] In another preferred embodiment, the disinfectant composition kills Staphylococcus aureus.

[0025] In a second aspect, the present invention provides a solid disinfectant powder, comprising:

[0026] Compound (I) 0.01–5 parts by weight,

[0027] 1 to 10 parts by weight of benzalkonium chloride.

[0028] In another preferred embodiment, the solid disinfectant powder comprises:

[0029] 0.5 to 2 parts by weight of compound (I),

[0030] 4-5 parts by weight of benzalkonium chloride.

[0031] In another preferred embodiment, the solid disinfectant powder is dissolved in water to prepare a solution of the compound of formula (I) with a concentration of 0.05-0.2 wt%, for use in air disinfection.

[0032] A third aspect of the present invention provides a method for air disinfection, comprising the steps of:

[0033] Disinfection is carried out by applying a disinfectant as described in the first aspect of the present invention to the space to be disinfected, or by applying an aqueous solution of a solid disinfectant powder as described in the second aspect of the present invention.

[0034] In another preferred embodiment, the amount of disinfectant used in the space is 1–20 mL / m². 3 .

[0035] In another preferred embodiment, the amount of disinfectant used in the space is 5–15 mL / m². 3 The optimal concentration is 8–12 mL / m 3 .

[0036] In another preferred embodiment, the weight percentage of the compound of formula (I) in the aqueous solution is 0.05-0.2 wt%.

[0037] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Detailed Implementation

[0038] Through extensive and in-depth research, and after numerous experiments and screenings, the inventors unexpectedly discovered for the first time that a combination of allicin E and benzalkonium chloride can effectively suppress the pungent odor of allicin E and effectively kill environmental pathogens even at very low dosages, demonstrating excellent disinfection effects. Based on this, the inventors completed this invention.

[0039] the term

[0040] To facilitate understanding of the invention, certain technical and scientific terms are specifically defined below. Unless otherwise expressly defined herein, all other technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art to which this invention pertains. Before describing the invention, it should be understood that the invention is not limited to the specific methods and experimental conditions described, as such methods and conditions can vary. It should also be understood that the terminology used herein is intended only to describe particular embodiments and is not intended to be restrictive; the scope of the invention will be limited only by the appended claims.

[0041] As used herein, the term “comprising” or its variations such as “including” or “comprising” are to be understood as including the said element or component without excluding other elements or other components.

[0042] The term “about” can refer to a value or composition within an acceptable margin of error for a particular value or composition as determined by a person skilled in the art, depending in part on how the value or composition is measured or determined. For example, as used herein, the expression “about 100” includes all values ​​between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0043] As used herein, unless otherwise stated, any concentration range, percentage range, proportion range, or integer range shall be understood to include any integer value within the range and, where appropriate, its fractional value (e.g., one-tenth and one-hundredth of an integer).

[0044] As used herein, the term “and / or” refers to and covers any and all possible combinations of one or more of the related listed items.

[0045] benzalkonium chloride

[0046] Benzalkonium chloride is a quaternary ammonium-based cationic surfactant, a heterogeneous mixture of various even-numbered alkyl chain alkylbenzyl chlorides. Its structural formula is as follows:

[0047] Garlic E

[0048] ALE (Allicin E) is a single compound synthesized biomimetically by modifying the structure outside the antibacterial active group of ordinary allicin. Its structure is shown in the following formula:

[0049] Garlic E has broad-spectrum bactericidal and green safety characteristics, and can effectively replace traditional antibiotics, supporting environmental disinfection, animal and plant protection, preservation, and upgrading of the biopharmaceutical industry.

[0050] The air disinfection agent of the present invention

[0051] In this invention, the terms "air disinfection preparation of the present invention", "disinfectant composition of the present invention" and "disinfectant of the present invention" are used interchangeably and all refer to a disinfection composition comprising 0.01 to 5 parts by weight of compound of formula (I), 1 to 10 parts by weight of benzalkonium chloride, and the balance being pure water.

[0052] The disinfectant of this invention contains garlic E, which has good biosafety, low toxicity, and good bactericidal effect.

[0053] In a preferred embodiment, the disinfectant comprises: 0.1 to 3 parts by weight of compound of formula (I), 1 to 8 parts by weight of benzalkonium chloride, and the remainder being pure water.

[0054] In a preferred embodiment, the disinfectant comprises: 0.5 to 2 parts by weight of compound of formula (I), 2 to 6 parts by weight of benzalkonium chloride, and the remainder being pure water.

[0055] In a preferred embodiment, the disinfectant comprises the following components: 0.05-0.2 wt% of a compound of formula (I); 0.1-1.0 wt% of benzalkonium chloride; and the balance being pure water.

[0056] In a preferred embodiment, the disinfectant comprises the following components: 0.08-0.12 wt% of a compound of formula (I); 0.4-0.5 wt% of benzalkonium chloride; and the balance being pure water.

[0057] In a preferred embodiment, the disinfectant has a sterilization rate of ≥95%.

[0058] In a preferred embodiment, the LD50 of the disinfectant 50 >5000 mg / kg·bw.

[0059] In a preferred embodiment, the LC of the disinfectant 50 >10000mg / m 3 .

[0060] In a preferred embodiment, the disinfectant kills Staphylococcus aureus.

[0061] The main advantages of this invention include:

[0062] 1. The disinfectant of this invention has good biosafety, low toxicity, and low LD50. 50 >5000 mg / kg·bw, LC 50 >10000mg / m 3 .

[0063] 2. The disinfectant of the present invention has a good bactericidal effect, with a bactericidal rate of ≥95%.

[0064] 3. The disinfectant formula of the present invention is simple, the ingredients are safe, and it does not damage chromosomes in the body.

[0065] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.

[0066] Preparation example of garlic E disinfectant solution

[0067] The garlic E-ingredient disinfectant of the present invention has the components shown in Table 0:

[0068] Table 0

[0069] Garlic E disinfectant solution was prepared by compounding according to the prescription shown in Table 0.

[0070] Neutralizer Identification Test in Example 1

[0071] 1.1 Method

[0072] The prepared garlic E disinfectant solution was used as the test sample, and the test was conducted according to the method in Appendix A of GB27948-2020 "General Requirements for Air Disinfectants". Test strain: Staphylococcus albus (8032), the 4th generation fresh slant culture was used for the test, and the strain was provided by BioVector NTCC Collection Center. Neutralizing agent: 2 times disinfectant to neutralize broth. The test groups were: (1) nutrient agar plate containing neutralizing agent + bacterial suspension; (2) (disinfectant + nutrient agar plate containing neutralizing agent) + bacterial suspension; (3) nutrient agar plate of the same batch + bacterial suspension, acted for 10 min, and the test was repeated 3 times.

[0073] 1.2 Results

[0074] The average number of bacteria in group 1 was 1.06 × 10⁻⁶. 3 CFU / mL, the average bacterial count in group 2 was 8.70 × 10⁻⁶. 2 CFU / mL, the average bacterial count in group 3 was 1.20 × 10⁻⁶. 3 The error rates for CFU / mL colony counts in groups 1, 2, and 3 were 10.54%, 10.31%, and 13.57%, respectively. The group numbers and their corresponding contents listed in the table are the same as those in GB 27948-2020 "General Requirements for Air Disinfectants".

[0075] Table 1. Results of the identification test for the neutralizing agent of Staphylococcus aureus (8032)

[0076] Note: The negative control showed no sterile growth.

[0077] Neutralizing agent identification tests showed that a 2x concentration of disinfectant effectively neutralized the bactericidal effect of the disinfectant of this invention on Staphylococcus albus (8032). Negative effects of the neutralizing agent and its products on Staphylococcus albus (8032) and the culture medium were not observed. This result indicates that the disinfectant of this invention has minimal impact on non-pathogenic microorganisms, thus making it a safe and effective disinfectant product.

[0078] Example 2: Simulation of on-site test of air disinfection effect

[0079] 2.1 Method

[0080] According to section 2.1.3.4 of the "Disinfection Technical Specifications" (2002 edition), an on-site simulation test of the air disinfection effect of the disinfectant of this invention was conducted. Staphylococcus aureus (8032) was used, and its fourth-generation fresh slant culture was used for the test. The strain was provided by the BioVector NTCC Collection Center. Neutralizing agent: 2 times the amount of disinfectant to neutralize the broth. Aerosol chamber: 20m 3 , 2 rooms.

[0081] The specific experimental steps are as follows:

[0082] 1. Inoculation: Take the test bacterial suspension and dilute it to the required concentration with nutrient broth culture medium. Spray while stirring. After spraying, continue stirring with a fan for 5 minutes, and then let it stand for 5 minutes.

[0083] 2. Sampling: Simultaneously, both the control and experimental groups were sampled before disinfection using a six-stage sieve air impactor sampler. The sampler was placed 1 m above the center of the chamber, and sampling was conducted at a flow rate of 28.3 L / min. This served as the control group and a positive control (i.e., the amount of contaminating bacteria) before the start of the experiment and before disinfection treatment in the experimental group. The sampling time for the control group was 20 s, and the sampling times for the experimental group before and after disinfection treatment were 20 s and 5 min, respectively.

[0084] 3. Disinfection: Disinfect the aerosol chamber of the experimental group for 60 minutes according to the instructions. Simultaneously, treat the aerosol chamber of the control group with the same method (without disinfectant). After the specified time, sample the aerosol chambers of both groups using the aforementioned method.

[0085] 4. After sampling, the sampling plates were incubated at 37.0℃ for 48 hours, and the results were observed and the number of colonies was counted.

[0086] 5. Negative control: 1.0 mL each of unused culture medium from the same batch, sampling solution, and PBS were incubated simultaneously with the above samples. The experiment was repeated three times, and the kill rate was calculated for each iteration.

[0087] 2.2 Results

[0088] Table 2 Results of simulated field test on air disinfection effect

[0089] Note: The negative control showed no sterile growth.

[0090] The above results indicate that at 20m 3 In a confined space, under experimental conditions, the air disinfectant was applied at a concentration of 10 mL / m². 3The concentration of the spray was applied to this enclosed space for 60 minutes, and the kill rate of Staphylococcus aureus in three tests was >99.90%, which meets the disinfection qualification requirements of GB 27948-2020 "General Requirements for Air Disinfectants".

[0091] Example 3: Field test of disinfectant for air disinfection

[0092] 3.1 Method

[0093] According to section 2.1.3.5 of the "Disinfection Technical Specifications" (2002 edition), a field test was conducted on the air disinfection effect of the disinfectant solution of this invention. Neutralizing agent: 2 times the amount of disinfectant solution to neutralize the broth. Room volume: 30m³. 3 .

[0094] The specific steps are as follows: Before treatment, use a six-stage sieve air impactor to sample naturally occurring bacteria in the air as a pre-disinfection sample (positive control); during the experiment, the sample mixture is diluted at the required dose of 10 mL / m³. 3 The concentration of the solution was atomized into the air. After 60 minutes of incubation, nutrient agar plates containing the corresponding neutralizing agent were placed in an impactor sampler, and air samples were taken from the same sampling point. During sampling, the six-mesh air impactor sampler was placed 1 m above the center of the room, with one sampling point set up. Sampling was performed at a flow rate of 28.3 L / min for 5 minutes, and the experiment was repeated three times. After sampling, the plates were incubated at 37.0℃ for 48 hours, and the total colony count and mortality rate were calculated.

[0095] 3.2 Results

[0096] Table 3. Results of field tests on disinfection effectiveness

[0097] Note: The negative control showed no sterile growth.

[0098] The above results indicate that at 30m 3 In the room, under experimental conditions, the air disinfectant was applied at a concentration of 10 mL / m². 3 The concentration was atomized into the air, and after 60 minutes of action, the elimination rate of naturally occurring bacteria in the air was >90% in three tests, which meets the requirements of GB 27948-2020 "General Requirements for Air Disinfectants".

[0099] Example 4 Acute oral toxicity test

[0100] 4.1 Methods

[0101] The mice were KM mice, a total of 20, half male and half female, weighing 18-22g, and were obtained from Jinan Pengyue Experimental Animal Breeding Co., Ltd.

[0102] Test basis: Acute oral toxicity test 2.3.1 of the "Disinfection Technical Specifications" (2002 edition).

[0103] Sample preparation: Weigh 5.00g of sample and add pure water to prepare 20mL of sample solution. Mix well and label for later use (final concentration is 250mg / mL).

[0104] Test method:

[0105] Animal preparation: KM mice were fasted overnight before the experiment, but their water intake was not restricted.

[0106] Method of administration: One maximum limit test, administered by gavage at a dose of 5000 mg / kg bw, with a gavage volume of 0.2 mL / 10 g bw.

[0107] Symptom observation: After exposure, KM mice were observed for signs of poisoning, the number of deaths, and the time of death. Autopsies were performed on dead animals and those euthanized at the end of the observation period. Abnormal tissues or organs were observed visually and further histopathological examinations were conducted. The observation period was 14 days.

[0108] Toxicity evaluation of disinfectants: LD50 50 LD50+ is practically non-toxic; 50 Those with a concentration of 501 mg / kg to 5000 mg / kg body weight are considered to have low toxicity; LD50 50 A dose of 51 mg / kg to 500 mg / kg body weight is considered moderately toxic; LD50 50 A dose of 1 mg / kg to 50 mg / kg body weight is considered highly toxic; LD50 50 Less than 1 mg / kg of body weight is considered highly toxic.

[0109] 4.2 Results

[0110] Table 4 Results of Acute Oral Toxicity Tests

[0111] No abnormal symptoms or deaths were observed in KM mice within 14 days of exposure, and gross necropsy examinations of the test animals after the observation period revealed no abnormalities. The acute oral LD50 of this sample in KM mice... 50 >5000 mg / kg·bw.

[0112] The above results indicate that the acute oral LD50 of the air disinfectant in KM mice is [not specified]. 50 The concentration of >5000 mg / kg·bw indicates that the acute oral toxicity test shows no practical toxicity, which meets the requirements of the "Disinfection Technical Specifications" (2002 edition).

[0113] Example 5 Acute Inhalation Toxicity Test

[0114] 5.1 Method

[0115] The mice were KM mice, a total of 20, half male and half female, weighing 18-22g, and were obtained from Jinan Pengyue Experimental Animal Breeding Co., Ltd.

[0116] Test basis: Acute inhalation toxicity test 2.3.2 of the "Disinfection Technical Specifications" (2002 edition).

[0117] Sample preparation: Collect 7.3 mL of sample; no preparation is required.

[0118] Sample specific gravity: Accurately transfer 6 1.0 mL test samples, weigh them and calculate their density. The average density is 1.01 g / mL and the relative density d (specific gravity) is approximately 1.01.

[0119] Test method:

[0120] Animal grouping: 20 KM mice were randomly divided into 4 groups, 2 males and 2 females.

[0121] Disinfection Method: Static disinfection was used. The static disinfection cabinet in this laboratory has a volume of 60L. According to section 2.3.2 of the "Disinfection Technical Specifications" (2002 edition), each disinfected mouse should be exposed to at least 3L of air per hour. This cabinet can hold 10 mice for 2 hours of disinfection. Four boxes (20 mice) were placed in the static disinfection cabinet in two separate sessions, each lasting 2 hours. Before disinfection, the animals were weighed. After placing the animals in the cabinet, 0.63mL of the test substance was added to the evaporation pan through the administration port. The evaporation temperature was set to 45℃ to allow it to evaporate, resulting in an air disinfectant concentration of 10605mg / m³. 3 (See the formula below for concentration calculation.)

[0122] In the formula, C: concentration of toxicity (mg / m³) 3 ); a: Disinfectant dosage (mL); V: Contamination cabinet volume (L); d: Disinfectant specific gravity.

[0123] Symptom observation: Observe and record the symptoms and mortality of KM mice during the exposure process and observation period. Perform autopsies on dead animals and animals euthanized at the end of the observation period for gross observation. If abnormal tissues or organs are found, further histopathological examination will be conducted. The observation period is 14 days.

[0124] LC 50 Calculation method: If the animal does not die within 14 days, LC is determined. 50 Value greater than 10000 mg / m 3 .

[0125] Toxicity evaluation of disinfectants: LC 50 2h greater than 10000mg / m 3It is practically non-toxic; LC 50 1001 mg / m² over 2 hours 3 ~10000mg / m 3 It is classified as low toxicity; LC 50 101 mg / m² over 2 hours 3 ~1000mg / m 3 This is classified as moderately toxic; LC 50 10 mg / m² over 2 hours 3 ~100mg / m 3 This substance is highly toxic; LC 50 2h less than 10mg / m 3 It is highly poisonous.

[0126] 5.2 Results

[0127] Table 5 Results of Acute Inhalation Toxicity Tests

[0128] The results showed that KM mice did not exhibit any abnormal symptoms or die within 14 days of exposure, and gross necropsy examinations of the animals after the observation period revealed no abnormalities. The LC50 of this sample for acute inhalation toxicity in KM mice was [not specified]. 50 >10605mg / m 3 .

[0129] The results showed that the acute inhalation toxicity LC50 of the air disinfectant in KM mice was [not specified]. 50 >10000mg / m 3 The acute inhalation toxicity test showed that it was practically non-toxic, meeting the requirements of the "Disinfection Technical Specifications" (2002 edition).

[0130] Example 6 Acute Eye Irritation Test

[0131] 6.1 Method

[0132] An acute eye irritation test was conducted on the disinfectant solution of this invention according to section 2.3.4 of the "Disinfection Technical Specifications" (2002 edition).

[0133] The specific methods are as follows: Animal preparation: Before the experiment, examine both eyes of each New Zealand rabbit. If no abnormalities are found, the rabbit can be used for the experiment. Administration method: Take 0.1 mL of the test substance and instill it into the conjunctival sac of the left eye of the animal. Passively close the eye for 4 seconds, and rinse with physiological saline after 30 seconds. The right eye is treated with physiological saline as a normal control.

[0134] Symptom observation: The damage and recovery of the conjunctiva, iris, and cornea of ​​the New Zealand rabbits were observed visually at 1 hour, 24 hours, 48 ​​hours, and 72 hours after eye drops. The experiment was terminated if no irritation reaction was observed within 72 hours.

[0135] Evaluation criteria: Acute irritation responses of the cornea, iris, and conjunctiva of New Zealand rabbits were scored according to Table 6. The "average score" for each animal at three different observation times (24h, 48h, and 72h) was calculated for corneal damage, iris damage, conjunctival hyperemia, and conjunctival edema (i.e., the sum of the scores for each animal at 24h, 48h, and 72h divided by the number of observations, 3). The intensity of the irritant effect of the test substance on the New Zealand rabbit's eyes was determined according to Table 7, based on the average scores and recovery time of corneal, iris, and conjunctival hyperemia and edema.

[0136] Table 6 Scoring criteria for acute eye irritation

[0137] Table 7 Grading Criteria for Eye Irritation Response

[0138] Note: Complete recovery refers to the animal's eye stimulation response score: corneal damage = 0, iris damage = 0, conjunctival hyperemia = 0 or conjunctival edema = 0 or 1.

[0139] **Irritation:** The reversible inflammatory response that occurs after contact with the test substance.

[0140] ***Corrosivity: Irreversible tissue damage caused upon contact with the test substance.

[0141] 6.2 Results

[0142] No corneal damage, iris damage, conjunctival hyperemia, or conjunctival edema were observed at 24h, 48h, and 72h after eye drops. Eye irritation response scores are shown in Table 8.

[0143] Table 8 New Zealand Rabbit Eye Stimulation Scores

[0144] Note: The average score is the sum of the scores at 24h, 48h, and 72h divided by the number of observation periods, 3.

[0145] Under the experimental conditions, the disinfectant solution of this invention caused non-irritating damage to the eyes of New Zealand rabbits, which meets the requirements of the "Disinfection Technical Specifications" (2002 edition).

[0146] Example 7 Mouse bone marrow polychromatic erythrocyte micronucleus test

[0147] 7.1 Method

[0148] 1. Testing basis: Disinfection Technical Specifications (2002 edition) 2.3.8.4 Mouse bone marrow polychromatic erythrocyte micronucleus test.

[0149] 2. Preparation of the test substance:

[0150] High-dose group (dose of 5000 mg / kg·bw): Weigh 5.0 g of sample and add pure water to prepare 20 mL of sample solution. Mix well and label for later use (final concentration of 250 mg / mL).

[0151] Medium-dose group (dose of 2500 mg / kg·bw): Take 5 mL of the high-dose group sample solution, add pure water to prepare 10 mL of sample solution, mix well, and label for later use (final concentration of 125 mg / mL).

[0152] Low-dose group (dose of 1000 mg / kg·bw): Take 2 mL of the high-dose group sample solution, add pure water to prepare 10 mL of sample solution, mix well, and label for later use (final concentration of 50 mg / mL).

[0153] Preparation was carried out continuously for two days.

[0154] 3. Test methods:

[0155] 3.1 Animal preparation: Thirty KM mice were selected and randomly divided into three dosage groups: high, medium, and low dose groups. Each group consisted of 10 mice, half male and half female.

[0156] 3.2 Dosage design: The median lethal dose (LD50) of this product in KM mice is greater than 5000 mg / kg·bw. Therefore, the high dose is designed to be 5000 mg / kg·bw, the medium dose is half of the high dose, and the low dose is one-fifth of the high dose.

[0157] 3.3 Method of administration: The administration method was 30-hour oral gavage, with a 24-hour interval between two administrations. Samples were collected 6 hours after the second administration. The gavage volume was 0.2 mL / 10 g·bw.

[0158] 3.4 Animals were euthanized by cervical dislocation. The femur was harvested, and the medullary cavity was flushed with calf serum. A smear of the flushing solution was prepared, fixed, stained with Giemsa, and examined under a microscope. The number of PCEs, NCEs, and PCEs containing micronuclei was observed and counted. A total of 1000 PCEs were counted for each animal (when counting 200 PCEs, the observed NCEs were also counted).

[0159] 3.5 Negative and positive control groups were set up: 20 mice, half male and half female, were used. The positive control group was given cyclophosphamide (40 mg / kg·bw), and the negative control group was given the test substance solvent. The procedures for the positive and negative control groups were the same as those for the experimental group.

[0160] 3.6 Evaluation Regulations

[0161] In the negative control group mice, the micronucleus cell rate generally did not exceed 0.3%.

[0162] Statistical analysis was performed using the Poisson distribution u-test. When the increase in micronucleated cell rate in each dose group compared to the negative control group was statistically significant and a dose-response relationship was observed, or when only one dose group showed a significant increase in micronucleated cell rate, and this was confirmed by repeated experiments, the test substance was deemed to have an in vivo chromosomal damage effect.

[0163] 7.2 Results

[0164] The experimental results showed that there were no significant differences between the low, medium, and high dose groups and the negative control group (P>0.05), while there was a significant difference between the positive control group and the negative control group (P<0.01). The results are shown in Table 9.

[0165] Table 9 Results of mouse bone marrow polychromatic erythrocyte micronucleus assay

[0166] Note: **P<0.01, compared with the negative control group.

[0167] Under the experimental conditions, the disinfectant of this invention did not cause an increase in the micronucleus rate in the bone marrow of KM mice, therefore it did not have an in vivo chromosomal damage effect, was mutagenic negative, and met the requirements of the "Disinfection Technical Specifications" (2002 edition).

[0168] in conclusion

[0169] 1. Neutralizing agent identification test shows that 2 times disinfectant solution can effectively neutralize the bactericidal effect of the disinfectant solution of the present invention on Staphylococcus albus (8032). The neutralizing agent and the neutralization product have no adverse effects on Staphylococcus albus (8032) and culture medium.

[0170] 2. Simulation results of field tests on air disinfection effects show that at a depth of 20m... 3 In a confined space, under experimental conditions, the disinfectant solution of the present invention was used at a concentration of 10 mL / m². 3 The concentration of the spray was applied to this enclosed space for 60 minutes, and the kill rate of Staphylococcus aureus in three tests was >99.90%, which meets the disinfection qualification requirements of GB 27948-2020 "General Requirements for Air Disinfectants".

[0171] 3. Field test results of air disinfection effect show that at 30m 3 In the room, under experimental conditions, the disinfectant solution of the present invention was used at a concentration of 10 mL / m². 3 The concentration was atomized into the air, and after 60 minutes of action, the elimination rate of naturally occurring bacteria in the air was >90% in three tests, which meets the requirements of GB 27948-2020 "General Requirements for Air Disinfectants".

[0172] 4. Acute oral toxicity LD50 of disinfectant in KM mice 50The concentration of >5000 mg / kg·bw indicates that the acute oral toxicity test shows no practical toxicity, which meets the requirements of the "Disinfection Technical Specifications" (2002 edition).

[0173] 5. Acute inhalation toxicity (LC50) of disinfectant in KM mice 50 >10000mg / m 3 The acute inhalation toxicity test showed that it was practically non-toxic, meeting the requirements of the "Disinfection Technical Specifications" (2002 edition).

[0174] 6. The disinfectant caused non-irritating damage to the eyes of New Zealand rabbits, which meets the requirements of the "Disinfection Technical Specifications" (2002 edition).

[0175] 7. The disinfectant does not cause an increase in the micronucleus rate in the bone marrow of KM mice, therefore it does not have an in vivo chromosomal damage effect, is mutagenic negative, and meets the requirements of the "Disinfection Technical Specifications" (2002 edition).

[0176] In summary, the disinfectant of the present invention is a safe and effective disinfectant composition. Although it uses garlic E, a component with a garlic odor, its compound formulation ensures that it is odorless in the environment, suitable for air disinfection, and has very good safety for organisms.

[0177] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A disinfectant composition characterized in that, include: 0.01 to 5 parts by weight of the compound of formula (I), 1-10 parts by weight of benzalkonium chloride; and The remaining amount of pure water.

2. The disinfectant composition of claim 1, wherein, The disinfectant composition comprises: 0.1 to 3 parts by weight of compound (I), 1 to 8 parts by weight of benzalkonium chloride, and the balance being pure water.

3. The disinfectant composition of claim 1, wherein The disinfectant composition comprises: 0.5 to 2 parts by weight of compound (I), 2 to 6 parts by weight of benzalkonium chloride, and the balance being pure water.

4. The disinfectant composition of claim 1, wherein The disinfectant composition comprises the following components: 0.05-0.2 wt% of compound of formula (I); 0.1-1.0 wt% benzalkonium chloride; and The remaining amount of pure water.

5. The disinfectant composition of claim 4, wherein The disinfectant composition comprises the following component: 0.08-0.12 wt% of a compound of formula (I); 0.4-0.5 wt% benzalkonium chloride; and The remaining amount of pure water.

6. The disinfectant composition of claim 1, wherein The disinfectant composition is an air disinfectant.

7. A solid disinfectant powder, characterized in that, include: 0.01 to 5 parts by weight of the compound of formula (I), 1 to 10 parts by weight of benzalkonium chloride.

8. The disinfectant powder of claim 7, wherein include: 0.5 to 2 parts by weight of a compound of formula (I), 4-5 parts by weight of benzalkonium chloride.

9. A method of air disinfection, characterized in that, Including the following steps: Disinfection is carried out by applying the disinfectant as described in any one of claims 1-6, or by applying an aqueous solution of the solid disinfectant powder as described in any one of claims 7-8, to the space to be disinfected.

10. The method of claim 9, wherein, The amount of the disinfectant used in the space is 1 to 20 mL / m 3 .

11. The method of claim 9, wherein, The disinfectant is used in an amount of 5-15 mL / m 3 .

12. The method of claim 9, wherein, The disinfectant is used in an amount of 8-12 mL / m 3 .

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