JP2023028568A - Agent for disinfection and virus inactivation - Google Patents

Abstract

To provide an agent for disinfection and virus inactivation that allows any kind of water to be blended therein without limitation, has a reliable and safe image, and contains a pH modifier with high pH stability.SOLUTION: An agent for disinfection and virus inactivation contains citrus seed extract, an amino acid or a salt thereof, and water, and has a pH of 8.0 or more.SELECTED DRAWING: None

Description

FIELD OF THE DISCLOSURE The present disclosure relates to disinfecting and virus inactivating agents.

For example, as disclosed in Patent Document 1, a sterilization/virus deactivator has been proposed that contains grapefruit seed extract as an active ingredient and has an alkaline pH. In this type of sterilization/virus inactivator, since the pH affects the efficacy, it is required to maintain the pH stably for a long period of time.

On the other hand, since disinfectants and virus inactivators used in general households are required to have a safe and secure image, it is easy for consumers to understand that the pH adjuster that is added to them is also safe. For example, it is preferably a substance used as a food additive.

In this regard, the sterilization/virus inactivating agent of Patent Document 1 uses a buffering agent composed of sodium carbonate and sodium bicarbonate, thereby making it possible to maintain a suitable pH for a long period of time.

Japanese Patent Application Laid-Open No. 2021-059512

By the way, the sterilization/virus deactivator of Patent Document 1 is an aqueous solution and is a water-based formulation. As a result of investigation by the inventors of the present application, there is no problem when using purified water (ion-exchanged water) as water, but it is clear that precipitation may occur when carbonate is added when tap water is used. became. Therefore, the sterilization/virus deactivator of Patent Document 1 has a problem that the water that can be blended is limited, and cheap tap water cannot be used. For the same reason, when the disinfectant/virus deactivator of Patent Document 1 is used in a plumbing area (such as a kitchen sink) in areas with extremely hard water, there is a possibility that deposits will form where it is used.

The present disclosure is in view of this point, there is no restriction on the water that can be blended, it has a safe and secure image, and it contains a pH adjuster that has excellent long-term pH stability. The purpose is to provide an agent.

In order to achieve the above object, the disinfectant/virus deactivator according to the present disclosure comprises a citrus seed extract such as grapefruit seed extract, a first amino acid or a salt thereof, and water such as tap water. have.

According to this configuration, the sterilization/virus deactivator is composed of a safe substance that is used as a food additive, so the image of safety and security can be easily conveyed to consumers. The water may be purified water (ion-exchanged water) or tap water. Whichever water is used, precipitation hardly occurs and the pH is stable over a long period of time. Therefore, high sterilization efficacy and virus inactivation efficacy can be obtained over a long period of time. Also, it can be used in areas with hard water (kitchen sinks, etc.) without deposits.

Also, the isoelectric point of the first amino acid can be 8 or higher. As a result, the pH of the sterilization/virus inactivating agent can be adjusted to 8.0 or higher to obtain high sterilization efficacy and virus inactivation efficacy. In addition, by setting the pH of the sterilization/virus deactivator to 9.0, the efficacy can be further enhanced, for example, sterilization can be performed in a short time and viruses can be inactivated. .

In addition, the sterilization/virus deactivator may further contain a pH stabilizer. A pH stabilizer can be, for example, a second amino acid that is different from the first amino acid. The dissociation constant derived from the _NH2 group of the second amino acid or salt thereof can be 8.0 or greater. By setting the dissociation constant to 8.0 or more, the sterilization/virus inactivating agent is stabilized at a pH at which high sterilization efficacy and virus inactivation efficacy can be obtained.

As described above, it contains citrus seed extract, amino acid, and water, and the pH is adjusted to 8.0 or higher, so there are no restrictions on the water that can be blended, and it has a safe and secure image. Moreover, the pH can be stabilized over a long period of time, and high sterilization efficacy and virus inactivation efficacy can be obtained. Also, it can be used around water in areas with hard water without generating sediments.

Hereinafter, embodiments of the present invention will be described in detail. It should be noted that the following description of preferred embodiments is essentially merely illustrative, and is not intended to limit the invention, its applications, or its uses.

A sterilization/virus deactivator according to an embodiment of the present invention contains a citrus seed extract, a first amino acid or a salt thereof, and water. The citrus seed extract and the first amino acid are also used as food additives, and the water is drinkable water. Therefore, the sterilization/virus deactivator is highly safe and can be used safely on tableware and the like in general households. The first amino acid or salt thereof preferably functions as a pH adjuster or pH stabilizer.

The disinfectant/virus inactivator is adjusted to pH 8.0 or higher. Specifically, the pH of the disinfecting/virus inactivating agent can be adjusted by the first amino acid or a salt thereof. In other words, the sterilization/virus deactivator contains the first amino acid or a salt thereof such that the pH of the sterilization/virus deactivator is 8.0 or higher. It can also be said that the sterilization/virus deactivator comprises an aqueous citrus seed extract solution containing the first amino acid or a salt thereof as a pH adjuster. That is, in this case, it can be said that the first amino acid or its salt functions as a pH adjuster. The pH of the disinfecting/virus inactivating agent can be set to a desired value depending on the type and content of the first amino acid or its salt, and may be 9.0 or higher, or 9.5 or higher. good too.

When blending the first amino acid as a pH adjuster, an amino acid with an isoelectric point (IEP) of 8 or higher can be used. Specific examples of the first amino acid in this case include arginine with an isoelectric point of 10.76 and lysine with an isoelectric point of 9.75. , an amino acid having an isoelectric point of 8 or more can be used as the first amino acid. Even when an amino acid having an isoelectric point of 8 or more other than arginine and lysine is used as the first amino acid, the action and effect of the disinfectant/virus inactivator are almost unchanged. The isoelectric point of the first amino acid may be 9 or higher, or 9.5 or higher. By using an amino acid with an isoelectric point of 8 or more as the first amino acid, a sufficient disinfecting effect against fungi and an inactivating effect against the non-enveloped feline calicivirus can be obtained. As the first amino acid, only one type of amino acid may be contained, or any two or more types of amino acids may be contained.

Moreover, a salt of the first amino acid can be blended as a pH adjuster. Specifically, salts such as sodium salts and potassium salts of amino acids that have a pH of 8 or higher after dissolution in water can be used. In this case, the pH after dissolving the salt may be within the above range, and the isoelectric point of the amino acid itself need not be 8 or higher.

The sterilization/virus deactivator preferably further contains a pH stabilizer for stabilizing the pH over a long period of time. "Long term" means, for example, 3 months or 6 months at room temperature. A pH stabilizer is an agent for stabilizing the pH so that the change in pH during this period is less than a predetermined value (for example, 1.0 or less). As mentioned above, an amino acid or a salt thereof can be used as a pH stabilizer. In addition, when the first amino acid (or its salt) is blended as a pH adjuster, a second amino acid (or its salt) different from the first amino acid (or its salt) is used as the pH stabilizer. be able to.

When blending an amino acid or a salt thereof as a pH stabilizer, for example, an amino acid having a dissociation constant (pKa2) derived from an NH ₂ group of 8.0 or higher can be used. In addition, although not essential, the pH should be 9.0 or higher or 9.5 or higher in order to exhibit high efficacy even if the contact time with the disinfectant/virus inactivator is short. is desirable. In this case, an amino acid having a dissociation constant derived from the _NH2 group of 9.0 or more, or 9.5 or more is used as a pH stabilizer. Examples of pH stabilizers include glutamic acid (pKa2 = 9.47), tryptophan (pKa2 = 9.41), glycine (pKa2 = 9.78), alanine (pKa2 = 9.87) and valine (pKa2 = 9.87). 74), leucine (pKa2 = 9.74), isoleucine (pKa2 = 9.76), cysteine (pKa2 = 10.70), tryptophan (pKa2 = 9.41), aspartic acid (pKa2 = 9.90), etc. Or these salts can be used. As the second amino acid, only one type of amino acid may be contained, or any two or more types of amino acids may be contained.

Substances other than amino acids (for example, sodium hydroxide, etc.) can also be used as pH adjusters. In this case, the first amino acid or salt thereof functions as a pH stabilizer, i.e. arginine, lysine, glutamic acid, tryptophan, glycine, alanine, valine, leucine, isoleucine, cysteine, tryptophan, aspartic acid, etc., and Any one or any two or more of these salts can be used.

Substances other than amino acids (for example, sodium salt of ethylenediaminetetraacetic acid) can also be used as pH stabilizers. In this case, the first amino acid or salt thereof can be one that functions as a pH adjuster, ie, arginine, lysine, or the like.

It is particularly preferred that both the pH adjuster and the pH stabilizer consist of amino acids or salts thereof. In this case, the sterilization/virus inactivator can be composed only of food ingredients (citrus seed extract and amino acids or their salts), making it highly safe, highly effective, and extremely practical. A bacteria/virus inactivator can be realized.

The upper limit of the pH of the sterilization/virus inactivator can be, for example, 11.5, and the contents of the first amino acid and the second amino acid can be set so that the pH is 11.5 or less. can. The upper limit of the pH of the disinfecting/virus inactivating agent may be pH 11.0. As a result, the sterilization/virus deactivator does not exhibit strong alkalinity, so safety during handling is enhanced.

The aqueous solution of citrus seed extract is obtained by dissolving citrus seed extract in, for example, tap water or ion-exchanged water. Tap water is water supplied by a general public water supply, and although the hardness of tap water differs somewhat depending on the region (region), tap water can be used anywhere without any problem. In this embodiment, tap water and ion-exchanged water can be mixed at any ratio and used.

Citrus seed extracts include, for example, grapefruit seed extract. The concentration of the grapefruit seed extract in the aqueous solution can be set within the range of 0.1% by mass or more and 5.0% by mass or less. Also, the lower limit of the concentration of the grapefruit seed extract is preferably 0.15% by mass, more preferably 0.2% by mass. Also, the upper limit of the concentration of the grapefruit seed extract is preferably 3.0% by mass, more preferably 0.8% by mass.

Grapefruit seed extract is extracted and purified from grapefruit seeds and is generally accepted as a food additive. When the grapefruit seed extract is obtained from grapefruit, the seeds are removed from the harvested grapefruit, the removed seeds are crushed, and the extract can be extracted from the crushed product. At this time, the grapefruit seed extract may be extracted from the pulverized material in an undried state, or the grapefruit seed extract may be extracted from the pulverized material in a freeze-dried state.

A solution such as water or alcohol can be used to extract the grapefruit seed extract. Examples of alcohol used as a solvent for extraction include ethanol. When extracting the grapefruit seed extract, the seeds may be heated, for example, to 30°C or higher. Grapefruit seed extract contains fatty acids, flavonoids, and the like. The grapefruit seed extract is preferably food grade, but it does not necessarily have to be food grade as long as safety is ensured.

Citrus seed extracts may be those other than grapefruit seed extracts, and seed extracts similarly extracted from citrus seeds such as lemon can also be used. A mixture of one citrus seed extract and another citrus seed extract may also be used. Even when a seed extract other than the grapefruit seed extract is used, the action and effect of the disinfectant/virus inactivator are almost the same.

The sterilization/virus deactivator may contain a component that produces carbonate ions when ionized or a component that produces hydrogen carbonate ions when ionized. Ampholytes such as (baking soda), sodium bicarbonate, sodium carbonate and the like can be mentioned. However, if tap water is used when these are blended, precipitation may occur as described above.

In addition, the sterilization/virus deactivator may contain a lower alcohol (specifically, ethanol). By containing alcohol, quick-drying property is improved, and it is possible to provide a bactericidal/virus inactivating agent with excellent usability. However, the disinfecting/virus inactivating agent of the present invention exhibits sufficient efficacy even if it does not contain alcohol. When alcohol is not contained, the sterilization/virus deactivator is extremely excellent in that it does not cause skin irritation or alcohol odor due to alcohol, and can be used safely even near infants.

The sterilization/virus deactivator can be used by accommodating it in a container having a spray lever and spraying it onto various articles (objects to be sterilized, objects to be virus inactivated). As the container having the spray lever, various conventionally used containers can be mentioned, and any container can be used. Alternatively, the disinfectant/virus deactivator can be sprayed by a spray device equipped with a hand-pump or an electric pump. In addition, the sterilization/virus inactivator can also be used by applying it to the article or by dripping it. The sterilization/virus inactivator can also be used by spraying it directly on, for example, cooking utensils such as cutting boards and kitchen knives, countertops, tableware, dishcloths, towels, and the like. The disinfectant/virus inactivator can also be used by spraying clothes, floors, walls, toilet bowls, washbasins, and interiors of automobiles. Alternatively, a nonwoven fabric or the like may be impregnated with a sterilizing/virus inactivating agent and provided in the form of a wet tissue. You can also spray the disinfectant/virus inactivator directly onto your hands.

(pH stability and presence or absence of precipitate)

Table 1 shows the components of the disinfecting/virus inactivating agents according to Examples 1 to 4, and the unit is % by weight. Arginine and sodium hydroxide are pH adjusters. In addition, sodium glutamate, EDTA.2Na (disodium salt of EDTA) is a pH stabilizer. That is, in any of the formulations of Examples 1 to 4, at least one of the pH adjuster and the pH stabilizer uses an amino acid (first amino acid).

Table 2 shows the results of the pH stability test and the presence or absence of precipitation in Examples 1-4. "pH after 1 week at 60°C" is the pH at the time of 1 week after the pH is adjusted and immediately stored in a heat-retaining cabinet at 60°C, and "pH after 4 weeks at 60°C" is the pH. It is the pH at the time when 4 weeks have passed since it was placed in a 60° C. heat-retaining chamber immediately after adjustment.

All of the formulations of Examples 1 to 4 exhibit excellent pH stability, with extremely small changes in pH of 0.7 or less from the initial pH. In addition, no sediment was observed after 4 weeks, even though tap water was used in all formulations of Examples 1-4. As Comparative Example 1, for example, as disclosed in Patent Document 1, even in an example in which the initial pH was set to 10 with sodium carbonate and sodium hydrogen carbonate, the pH stability was high, but the ion exchange A sediment was seen when tap water was used instead of water. In Comparative Example 2, when the initial pH was set to 11.96 with alkaline electrolyzed water and calcined shellfish calcium, the pH after 3 weeks at 60°C was 10.16, a large change of about 1.8. was taken.

In Examples 1 to 4, all of the water is tap water, but similar results can be obtained by using ion-exchanged water for all or part of the water.

(Trichophyton eradication efficacy test)
Example 2 shown in Table 1 was used as a test agent in the Trichophyton eradication efficacy test. The strain is Trichophyton mentagrophytes (NBRC32409). The test location is a sterile room and an incubation room set at 25°C.

First, by mixing 0.1 ml of the bacterial suspension adjusted to about 10 ⁹ cfu/ml and 10 ml of the test agent, the test agent (Example 2 or physiological saline) was added to the bacterial suspension for a predetermined time. (20 seconds, or 60 seconds). After a predetermined period of time, 1 ml of the mixture of the bacterial suspension and the test agent was sampled and added to 9 ml of the SCDLP liquid medium for dilution. Four dilution series were prepared, 200 μl of the diluted solution was spread on a nutrient agar medium, and the number of viable cells was measured. The results are shown in Table 3. In Example 2, the number of viable bacteria was reduced by two digits or more from the physiological saline solution (blank), and therefore the trichophytosis eradication rate in Example 2 was 99%. It should be noted that the test preparations of Examples 1, 3 and 4 exhibit similar sterilization rates against Trichophyton.

(Virus inactivation test)
The test agent used in the virus inactivation test is Example 2 shown in Table 1. The viruses tested were feline calicivirus and the so-called novel coronavirus SARS-CoV-2.

First, the results for feline calicivirus will be described. This feline calicivirus is being tested as an alternative to the structurally similar norovirus, which belongs to the same taxonomic family. This is because norovirus is difficult to culture and no method has yet been established for easily evaluating the infectivity titer. That is, it is generally believed that an agent that exhibits sufficient antiviral activity in tests using feline calicivirus is highly likely to exhibit sufficient antiviral activity against norovirus. The test method is as follows.

1. 10 μL of the virus solution is inoculated into 90 μL of the test agent (sterilized water in Example 2).
2. After 30 seconds, serially dilute the solution obtained in 1 above with DMEM (any FBS concentration).
3. 50 μl of the solution obtained by serially diluting in 2 above is inoculated to cells cultured on a cell culture microplate (96 wells).
4. After 1 hour, aspirate the liquid from each hole.
5. 100 μl of DMEM with an arbitrary FBS concentration is added dropwise and cultured for 4 days in a carbon dioxide incubator (CO ₂ concentration: 5%).
6. About 100 μl of ethanol:acetic acid=4:1 (weight ratio) is poured into each well for immobilization (about 40 minutes).
7. After removing the fixative, 30 μl of 45% ethanol containing 0.5% amido black and 30 μl of 10% acetic acid are added to stain the cells (for about 1 hour).
8. Immerse the plate in a container filled with water, and wash away the staining solution so as not to damage the cells (about 1 hour).

After confirming the viability of the cells with the naked eye, the 50% tissue culture infectious titer (TCID ₅₀ /ml) was calculated. The lower this value, the lower the infectivity. Then, the virus inactivation rate of Example 2 compared with sterilized water (blank) was determined. The results are shown in Table 4. As shown in Table 4, in Example 2, the feline calicivirus infectivity titer was reduced by four orders of magnitude or more from the blank, and therefore the inactivation rate of feline calicivirus in Example 2 was 99.99% or more. Met. It should be noted that the test agents of Examples 1, 3 and 4 exhibit similar inactivation rates against feline calicivirus.

In general, non-enveloped viruses such as feline calicivirus are more resistant to various disinfectants and antibacterial agents than enveloped viruses such as influenza virus. Therefore, agents that are effective against non-enveloped viruses are likely to be effective against enveloped viruses in a shorter period of time.

Next, a study using an isolate of the so-called novel coronavirus SARS-CoV-2, which is believed to cause the novel coronavirus infection (COVID-19), will be described. The test agent is Example 2.

1. First, cells are cultured in a monolayer in a cell culture microplate (96 wells) using a cell growth medium. Cells were VeroE6/TMPRSS2 cells and 100 μl of cell growth medium was used per well (well).
2. 90 μl of the test agent (Example 2 or sterilized water) is mixed with 10 μl of the virus solution and allowed to act for a predetermined time (10 seconds).
3. After the action, 0.05 ml is collected from the mixture and diluted 100-fold with the medium to stop the action.
4. A 10-fold serial dilution series is prepared in a cell maintenance medium, 50 μl of each is inoculated into monolayer cultured cells, and cultured for 3 to 4 days.

Cells were confirmed to be alive or dead by microscopic observation, and the 50% tissue culture infectivity titer (TCID ₅₀ /ml) was calculated by the Behrens-Karber method. The lower this value, the lower the infectivity. Then, the virus inactivation rate of Example 2 compared with sterilized water (blank) was determined.

As shown in Table 5, the sterilization/virus deactivator of Example 2 reduced the infectious titer of the novel coronavirus by more than three digits from the blank, despite the extremely short contact time of 10 seconds. Therefore, more than 99.9% of the new coronavirus could be inactivated.

(Efficacy test for various bacteria)
In addition, as a result of conducting an efficacy test against various bacteria, as shown in Table 6, the sterilization rate against multiple kinds of bacteria was 99.99% or more. The test method is as follows. 0.1 ml of the bacterial suspension adjusted to 10 ⁷ to 10 ⁹ cfu/ml for each bacterial solution was brought into contact with 10 ml of the test agent (Example 2 or physiological saline) for 20 seconds, and then 1 ml of the liquid was added to the SCDLP liquid medium. Added to 9 ml and diluted. Vibrio parahaemolyticus was the SCDLP liquid medium containing 3% sodium chloride. Then, it was diluted with the same solution as above, and the number of viable bacteria was measured.

Acinetobacter is Acinetobacter baumannii JCM6841. Campylobacter is Campylobacter jejuni subsp. Jejuni ATCC33560. E. coli (O-157) is Escherichia coli (E. coli, serotype O157:H7, verotoxin type I and II producing strain) ATCC43895. Legionella is Legionella pneumophila GIFU9134. The Listeria is Listeria monocytogenes VTU 206. Salmonella is Salmonella enterica subsp enterica NBRC3313. Staphylococcus aureus is Staphlococcus aureus NBRC12732. Vibrio parahaemolyticus is Vibrio parahaemolyticus RIMD2210100.

Table 7 shows the results of an efficacy test against Pseudomonas aeruginosa (NBRC13275). The test method is as follows. After contacting 0.1 ml of the bacterial suspension adjusted to about 10 ⁸ cfu/ml with 10 ml of the test preparation for 20 seconds, 1 ml of the liquid was added to 9 ml of the SCDLP liquid medium and diluted. After diluting with the same solution as above, 200 μl of the diluted solution was spread on a nutrient agar medium, and the number of viable bacteria was measured.

As shown in Table 7, the antibacterial/virus inactivating agent exerts a sufficient antibacterial effect even against Pseudomonas aeruginosa.

As described above, a composition containing a citrus seed extract, an amino acid, and water and having a pH of 8.0 or higher exhibits a high disinfecting effect, and can be called a disinfecting agent. In addition, a composition containing a citrus seed extract, an amino acid, and water and having a pH of 8.0 or higher exhibits a high virus-inactivating effect, and can be called a virus-inactivating agent. . In addition, the above composition can also be called a sterilizing agent having a virus-inactivating effect, or a virus-inactivating agent having a sterilizing effect.

(contaminant)
Next, contamination will be explained. The staining property can be determined by spraying the test agent onto, for example, a black object, drying it completely, and then visually observing whether or not any residual powder is observed. If powder residue is found, it can be determined that there is contamination, and if no powder residue is found, it can be determined that there is no contamination. Concerning staining, all of Examples 1 to 4 were free of staining.

(Sterilization method, virus inactivation method)
The treatment of attaching or contacting a composition containing a citrus seed extract, an amino acid, and water and having a pH of 8.0 or more to an object to be sterilized sterilizes the object to be sterilized as described above. It is a sterilization method because it can be used. In addition, the treatment of adhering or contacting a virus inactivation target with a composition containing a citrus seed extract, an amino acid, and water and having a pH of 8.0 or higher is effective for virus inactivation as described above. It is a virus inactivation method as it can inactivate viruses present in the object. In addition, since sterilization and virus inactivation can be performed at the same time, the treatment of adhering or contacting the composition to an article or the like can also be called a sterilization and virus inactivation method.

The time for which the composition is brought into contact with an article or the like can be, for example, 30 seconds or longer. This time can be changed depending on the type of bacteria or virus, and can be set to, for example, 120 seconds or more depending on the type.

(Action and effect of the embodiment)
As described above, according to the sterilization/virus deactivator according to this embodiment, a desired pH can be maintained for a long period of time while maintaining a safe and secure composition containing a citrus seed extract, an amino acid, and water. It can be kept stable and obtain high sterilization effect and virus inactivation effect.

The above-described embodiments are merely examples in all respects and should not be construed in a restrictive manner. Furthermore, all modifications and changes within the equivalent range of claims are within the scope of the present invention.

As described above, the disinfectant/virus inactivator according to the present invention can be used by spraying it on, for example, cooking utensils, countertops, tableware, dish towels, towels, and the like.

Claims (6)

citrus seed extract and
a first amino acid or a salt thereof;
containing water and
A disinfecting/virus inactivating agent with a pH of 8.0 or higher. In the sterilization / virus inactivating agent according to claim 1,
A disinfecting/virus inactivating agent with a pH of 9.0 or higher. In the sterilization / virus inactivating agent according to claim 1 or 2,
The disinfecting/virus inactivating agent, wherein the first amino acid has an isoelectric point of 8 or more. In the sterilization/virus deactivator according to any one of claims 1 to 3,
A disinfectant/virus inactivator further containing a pH stabilizer. In the sterilization / virus inactivating agent according to claim 4,
The pH stabilizing agent is a second amino acid different from the first amino acid or a salt thereof. In the sterilization/virus deactivator according to claim 5,
A disinfecting/virus inactivating agent, wherein the dissociation constant derived from the _NH2 group of the second amino acid or its salt is 8.0 or more.