JP7442833B2 - Antibacterial method - Google Patents

Description

The present invention relates to an antibacterial method, and particularly to a safe antibacterial method.

Antibacterial water generally contains antiseptic and sterilizing ingredients that inhibit the propagation and growth of microorganisms to prevent the contents and containers from deterioration such as putrefaction caused by microorganisms. Many antiseptic and sterilizing ingredients are listed on the list of ingredients that can be mixed in (positive list), so there are restrictions on their combination.

For example, it is an antibacterial water for wiping containing a disinfectant, which is used for purposes selected from the group consisting of sensitive skin, acne prevention, and acne improvement, and contains the following ingredients (A) and the following ingredients (B). Antibacterial water characterized by containing the following component (C), the following component (D), and the following component (E) is known (Patent Document 1).
Ingredient (A): Anti-inflammatory agent Ingredient (B): Disinfectant Ingredient (C): Fatty acid ester nonionic surfactant Ingredient (D): Humectant Ingredient (E): Water

JP2016-132623A

As described above, many of the conventional technologies including the above-mentioned Patent Document 1 contain sterilizers to prevent the contents and containers from deterioration such as putrefaction caused by microorganisms. On the other hand, many consumers still believe that these antibacterial ingredients are harmful, so it is desirable not to include preservatives and disinfectants. Therefore, it is desirable to suppress the propagation and growth of microorganisms by means other than antiseptic and sterilizing ingredients before use, so that the contents and containers do not undergo deterioration such as putrefaction due to microorganisms.

Therefore, an object of the present invention is to provide an antibacterial method that has antibacterial activity and is safe.

In order to achieve the above object, the present inventors conducted intensive studies on compositions that have antibacterial properties without incorporating preservative or sterilizing ingredients, and as a result, they discovered the present invention.

That is, the indoor antibacterial method of the present invention provides an antibacterial water composition containing at least one amino acid selected from L-cysteine, L-arginine, L-lysine, L-histidine, or salts thereof, An antibacterial water composition having a pH of 10.9 or more is sprayed indoors , and a piezoelectric vibrator or a two-fluid nozzle is used in the spraying to generate a mist. The present invention is characterized by comprising a step of suppressing floating viruses , and a step of antibacterializing the interior of the room by the suppression .

Further, in a preferred embodiment of the antibacterial method of the present invention, in the spraying step, the average particle diameter of the sprayed mist is 10 μm or less.

In a preferred embodiment of the antibacterial method of the present invention, the antibacterial water composition further contains reducing ion water.

Further, in a preferred embodiment of the antibacterial method of the present invention, the content of the amino acid is in the range of 1.0% by mass to 0.00001% by mass based on the total amount of the antibacterial water composition. shall be.

Further, in a preferred embodiment of the antibacterial method of the present invention, the content of the reducing ion water is in the range of 10% by mass to 90% by mass with respect to the total amount of the antibacterial water composition. do.

Further, in a preferred embodiment of the antibacterial method of the present invention, the antibacterial water composition is characterized in that it does not contain a pH adjuster.

According to the antibacterial method of the present invention, it is possible to provide an antibacterial water composition having antibacterial properties without adding so-called preservatives or bactericidal agents.

FIG. 1 is a diagram showing an example of the effect of the antibacterial water composition in one embodiment used in the method of the present invention. FIG. 2 is a diagram showing an example of the effect of the antibacterial water composition in one embodiment used in the method of the present invention. FIG. 3 is a diagram showing an example of the effect of the antibacterial water composition in one embodiment used in the method of the present invention. FIG. 4 is a diagram showing an example of the effect of an embodiment of the method of the present invention.

The antibacterial method of the present invention provides an antibacterial water composition containing at least one amino acid selected from L-cysteine, L-arginine, L-lysine, L-histidine, or salts thereof, the antibacterial water composition comprising: The method is characterized by comprising a step of spraying an antibacterial water composition having a pH of 10.9 or higher, more preferably 11.5 or higher. This is an attempt to provide an antibacterial method for consumers who do not want the addition of antibacterial ingredients by using a composition with antibacterial action that does not contain so-called preservatives or bactericidal agents.

In the present invention, since the alkalinity of the antibacterial water composition is extremely low, even if the antibacterial water composition is sprayed and comes into contact with the body, it will instantly become weakly acidic due to the skin's neutralizing ability. It can be used as non-irritating antibacterial water. In other words, the antibacterial water composition used in the antibacterial method of the present invention has a pH of 10.9 or higher to around pH 11.5, so it is an antibacterial water that has antibacterial properties even without the addition of preservatives, and should be applied to the skin. However, one of its characteristics is that it instantly becomes weakly acidic (pH around 6.0 on the skin surface).

In a preferred embodiment of the antibacterial method of the present invention, in the spraying step, the average particle diameter of the sprayed mist is 10 μm or less, from the viewpoint of sufficient diffusion in the space when performing antibacterial treatment in the space. More preferably, the thickness is 3 μm or less.

In a preferred embodiment of the antibacterial method of the present invention, in the spraying step, a piezoelectric vibrator or a two-fluid nozzle is used to sufficiently diffuse the antibacterial water composition into the space. It is characterized by generating mist.

In the present invention, the case where a piezoelectric vibrator is used to generate mist will be explained as follows. As long as it uses a piezoelectric vibrator, it is not particularly limited, but for example, a mist device leased from Sunny Place Co., Ltd. (product name: PLUME2, manufactured by Sunny Place Co., Ltd., for example, the mist device described in No. 5801778, etc.) ) can be used. The piezoelectric vibrator plates provided in the water storage tank can each vibrate at an operating frequency of 2.4MHz using an operating board. When the piezoelectric vibrator plate vibrates, the liquid in the water storage tank has energy, and it is possible to generate mist with various particle sizes, for example, mist with an average particle size of 3 μm or less. Note that the piezoelectric vibrator plate is configured to operate by receiving an operating voltage of 24V from the power supply unit of the main body of the device, and is capable of continuous spraying.

Additionally, with a single-fluid nozzle, the particles are large and may not be sufficiently dispersed within the room, causing the floor to become wet, but by using a two-fluid nozzle, particles with an average particle diameter of 10 μm can be made into fine particles. Therefore, it becomes possible to exhibit virus suppression performance. A two-fluid nozzle can refer to a nozzle that atomizes a liquid using a high-speed flow of compressed air or the like. It has the following features compared to single-fluid nozzles that spray using only a pump. 1. Excellent atomization performance (fluid nozzles can atomize particles with an average particle size of 10 μm or less, which is difficult with a single fluid nozzle); 2. Large turndown (The ratio between the minimum and maximum adjustable spray flow rate is called turndown ratio, and the spray flow rate can be adjusted over a wide range while keeping the particle size and flow distribution constant, making it suitable as a spray flow rate adjustment nozzle. 3. Large foreign matter passage diameter (compared to a single fluid nozzle with the same amount of water), 3. Large foreign matter passage diameter.

In the two-fluid nozzle, as the gas, in addition to compressed air, an inert gas (such as N ₂ ) or steam can be used.

Examples of the type of two-fluid nozzle include internal mixing type, external mixing type, and collision type, all of which are applicable to the present invention.

Note that the average particle diameter can be determined by a liquid immersion method. The immersion method is a method in which the mist is caught on a plate glass coated with a thick layer of silicone oil, a quickly enlarged photograph is taken, and the number of particles by size is counted from the resulting photograph.

Further, in a preferred embodiment of the antibacterial method of the present invention, the content of the amino acid is in the range of 1.0% by mass to 0.00001% by mass, more preferably 0% by mass, based on the total amount of the antibacterial water composition. It is characterized by a range of .01% by mass to 0.0001% by mass. The reason why this range is set is that an amount of this level can be effective as a composition having antibacterial properties.

In the antibacterial water composition applicable to the antibacterial method of the present invention, the pH value can be adjusted as appropriate by adjusting the type and amount of amino acids to be blended. In general, the pH can be adjusted using a pH adjuster including sodium carbonate, sodium hydrogen carbonate, etc. in addition to sodium carbonate, but the use of such a pH adjuster increases the alkalinity of the composition and makes it difficult to maintain the pH. The acidity becomes stronger, and when applied to the skin, it tends to be difficult to instantly return to a slightly acidic state.

Therefore, from this point of view, a preferred embodiment of the present invention is characterized in that it does not contain a pH adjuster.

In addition, in the present invention, the pH of the antibacterial water composition is set to be 10.9 or higher.With such a pH value, many favorable effects can be achieved without adding so-called antiseptic and sterilizing ingredients. This is because it is possible to eliminate alkaline microorganisms.

In a preferred embodiment of the antibacterial method of the present invention, the antibacterial water composition further contains reducing ion water. As mentioned above, the pH value can be adjusted as appropriate depending on the type of amino acid to be blended, the amount to be blended, etc. However, depending on the amino acid to be blended and the amount to be blended, there is a possibility that the desired pH value may not be achieved. In this case, reducing ionic water can be used. Reduced ion water can be electrolyzed and made into alkaline water. For example, examples of reduced ionized water include S-100 manufactured and sold by A.I. System Products Co., Ltd. S-100 is a high-performance reducing ionized water that has been electrolyzed and is labeled for cosmetics. The name is "water" and it is alkaline water with a pH of 12±0.5.

Further, in a preferred embodiment of the antibacterial method of the present invention, the content of the reducing ion water is in the range of 10% by mass to 90% by mass, more preferably 10% by mass, based on the total amount of the antibacterial water composition. It is characterized by a range of % by mass to 20% by mass.

An example of an antibacterial water composition applicable to the antibacterial method of the present invention will be described below using examples, but the present invention is not to be interpreted as being limited to the following examples.

Example 1
First, we tried an example of an antibacterial water composition that is alkaline and contains no antiseptic or sterilizing ingredients. Arginine was used as an example of an amino acid. Specifically, 1.0% by mass of L(+)-arginine (manufactured by Wako Pure Chemical Industries, Ltd.) was blended with 99.0% by mass of purified water to prepare an alkaline aqueous solution, and the pH was measured. In addition, for reference, 20% by mass of electroreducible ionized water S-100 (manufactured by A.I. System Products Co., Ltd.) was mixed with 80.0% by mass of purified water to create an alkaline aqueous solution and the pH was measured. (Reference example).

Note that the following models and electrodes were used for pH measurement.
pH meter model: pH METER F-71 (Horiba, Ltd.)
pH meter electrode: #9615-10D (Horiba Ltd.)

Example 2
Next, 20.0% by mass of S-100 and 1.0% by mass of L-arginine were blended with 79.0% by mass of purified water to prepare an alkaline aqueous solution, and the pH was measured in the same manner as in Example 1. .

Table 1 shows the pH measurements of Examples 1 and 2 and Reference Example.

From the results in Table 1, it was found that the reference example had the highest pH measurement value.

Examples 3 to 6
Based on the results in Table 1, various adjustment examples were created according to the same procedure as in Example 1. Table 2 shows component examples and adjustment examples of antibacterial water compositions applicable to the antibacterial method in one embodiment of the present invention.

Example 7
Next, an antibacterial water composition applicable to the antibacterial method of the present invention was prepared using L-histidine as the amino acid and various amounts of reduced ion water according to the procedure of the above example. did. The results are shown in Table 4.

As a result, it was found that a composition having a pH of 10.9 or higher can be prepared, and a composition having antibacterial properties can be prepared without containing so-called preservatives or bactericides. Furthermore, it has been found that if the same amounts of three types of basic amino acids are blended, 0.001% or less is preferable.

In addition, the present invention does not use pH adjusters such as sodium carbonate, sodium bicarbonate, etc. in addition to sodium carbonate, so it is possible to provide a composition with extremely low alkalinity, so when applied to the skin, it is possible to provide a composition with extremely low alkalinity. It has the advantageous effect of instantly becoming weakly acidic and not irritating the skin with its ability to neutralize the skin.

Example 8
Next, the antibacterial water composition applicable to the antibacterial method of the present invention was actually investigated for its virus inactivation effect using various viruses. Specifically, the virus inactivation effect was confirmed when the antibacterial water composition applicable to the antibacterial method of the present invention was reacted with influenza virus, feline calicivirus, and PED virus. Sterile phosphate buffer was used as a control material. Table 5 shows the components of the antibacterial water composition applicable to the present invention used in the virus inactivation test.

The test microorganisms are as follows.
・Influenza virus: swine influenza virus H1N1 IOWA strain, cultured cells: MDCK cells (canine kidney-derived cell line)
・Feline calicivirus: feline calicivirus F9 strain, cultured cells: CRFK cells (feline kidney-derived cell line) *Substitute for norovirus
・PED virus: Porcine epidemic diarrhea virus P-5V strain, cultured cells: vero cells (established cell line derived from African green monkey kidney epithelium) *Porcine infectious coronavirus (as a substitute for the new coronavirus)

Regarding the setting of test plots, etc., the test plots are as follows.
Treatment: Add 0.1mL of virus solution to 1mL of test material (antibacterial water composition applicable to the antibacterial method of the present invention) Sensitization time: 30 minutes after start of test

In the control area, the results are as follows.
Treatment: Add 0.1 mL of virus solution to 1 mL of phosphate buffer Sensitization time: 0 minutes, 30 minutes after start of test

The test method was carried out with reference to ``Virus Experimental Studies General Theory, Second Revised Edition, Maruzen Co., Ltd. Virus Neutralization Test Method.''

<Test procedure>
1) Preliminary exam:
Prior to the test, the effects of the test materials on cultured cells (cytotoxicity) were investigated. After serially diluting the test material 10 times with phosphate buffer, it was inoculated into cultured cells, and the highest concentration indicating the normal state of the cells after culture was confirmed, and the virus concentration to be used in the test was determined. As a result, no cytotoxicity was confirmed in MDCK cells, CRFK cells, or vero cells. For this reason, the concentration of each virus added was set to 10 ⁵ TCID ₅₀ /mL or more, and the detection limit was set to <10 ^1.5 TCID 50 /mL.

2) Main test/test solution mixture:
According to the test category, 1 mL each of the test materials and phosphate buffer was taken out, and the virus solution was added to the concentration determined in the preliminary test. After adding the virus solution, the mixture was allowed to stand at room temperature (25°C) for a predetermined period of time.

3) Main test/cell inoculation and bacterial count measurement:
The sensitized mixed solution for each test category was serially diluted 10 times, and 100 μL each was inoculated onto cells cultured in a 96-well plate. Judgment is made by culturing at 37°C and carbon dioxide (5%) for 5 days, then in the case of influenza virus, collect the culture supernatant in each well and check for the presence or absence of virus proliferation by hemagglutination reaction. The concentration was calculated. In the case of feline calicivirus and PED virus, the cultured cells were observed under a microscope, and the presence or absence of virus proliferation was confirmed by CPE (cytopathia) appearing in the cultured cells, and its concentration was calculated.

<Results>
1) Influenza virus The test results for influenza virus are shown in Figure 1. In the control group, no change in viral load was observed from the start of the test until 30 minutes (10 ^8.1 TCID ₅₀ /mL (130000000)). In the test area, the concentration was 10 ^3.9 TCID ₅₀ /mL (8000) (99.99% decrease) 30 minutes after the start.

2) PED virus
The test results for PED virus are shown in Figure 2. In the control group, no change in viral load was observed from the start of the test until 30 minutes (10 ^8.1 TCID ₅₀ /mL (130000000)). In the test area, the concentration was 10 ^3.9 TCID ₅₀ /mL (8000) (99.99% decrease) 30 minutes after the start.

3) Feline calicivirus The test results for feline calicivirus are shown in Figure 3. In the control group, no change in viral load was observed from the start of the test until 30 minutes (10 ^6.7 TCID ₅₀ /mL (5,000,000)). In the test area, the concentration was 10 ^1.9 TCID ₅₀ /mL (99.99% decrease) 30 minutes after the start.

This time, we conducted a test of the inactivation effect of the test material on influenza virus, feline calicivirus, and PED virus. As a result, the test material (antibacterial water composition applicable to the antibacterial method of the present invention) had a virus inactivation effect of 99.99% or more against influenza virus, feline calicivirus, and PED virus in a reaction time of 30 minutes or more. It turns out that there is something.

Example 9
Next, we conducted a test for antibacterial effects against indoor airborne viruses. Specifically, this test was conducted in a 25m3 space to evaluate the suppression performance against indoor airborne viruses (influenza virus) by operating test equipment (test material spray). The test viruses are as follows.

Test virus: Influenza virus: swine influenza virus H1N1 IOWA strain Cultured cells: MDCK cells (canine kidney-derived cell line)

<Test materials>
Test equipment: PLUME (The test equipment was used in continuous operation mode for 60 minutes. Test material: Amion (*The test material was used as an undiluted solution.) (Note that Amion is a composition having the components shown in Table 5 above. .

Table 6 shows the ward settings.

The test was conducted with reference to JEM1467 "Household Air Purifier" Appendix D "Indoor Airborne Virus Suppression Performance Evaluation Test".

<Virus liquid preparation method>
1) Thaw the virus solution cryopreserved at -70°C or below and inoculate it into MDCK cells.
2) After adsorbing the virus to the cells at 37°C for 1 hour, the inoculated virus solution was removed and the cells were washed twice with sterile PBS.
3) MEM (cell maintenance medium) was added and cultured at 37°C and 5% CO2 for 3 days until the conditions of 4) above were achieved.
4) When approximately 70 to 80% cytopathic effect (hereinafter referred to as CPE) was observed, the cultured virus solution was collected.
5) The collected culture supernatant was centrifuged at 3000 rpm for 30 minutes, and the supernatant was dispensed to serve as a test virus solution.

<Test procedure and method>
(1) Test environment A closed space of approximately 25 m3 (3.5 x 3.5 x 2.2 m) was used as the test environment. During the test, the temperature was 25°C ± 5°C, and the humidity at the start of the test was 50% RH.
(2) Method 1. To prepare for the test, 5 mL of the virus solution was sprayed into the test chamber, and after the spraying was finished, the room was stirred with a fan for 2 minutes.
2. In the control group, the start of the test was immediately after 5 mL of virus solution was sprayed into the test room, and airborne viruses in the room were collected over time according to the test settings while the internal stirring fan was running.
3. After installing and operating the test equipment, the test area was sprayed with virus solution in the same manner as the start of the test, and airborne viruses in the room were collected over time according to the test settings with the test equipment and stirring fan operating.
4. Viruses were collected using an impinger by sucking 10 L of room air into 10 mL of cell maintenance medium (MEM medium) at 5 L/min for 2 minutes. The MEM medium containing the virus was further serially diluted 10 times in MEM medium.
5. After inoculating MDCK cells with each dilution, they were cultured at 37°C under 5% CO2 for 5 days.
6. The virus titer (TCID50) was determined from the presence or absence of CPE and the presence or absence of chicken red blood agglutination reaction in the culture supernatant.

<Evaluation>
In the test results, the reduction rate (%) in the test area relative to the control area was calculated for each testing time point to confirm the effect. In addition, in this test, the reduction rate was calculated using the following formula.

Reduction rate (%) = 100 × (control area - test area) / control area

The test results for influenza virus are shown below. In the control group, a natural decline in the viral load was observed from the start of the test until 60 minutes later (104.5 → 104.1 TCID50/10Lair). In the test area, it was 104.1 TCID50/10Lair (59.3% decrease) 6 minutes after the start of the test, 103.7 TCID50/10Lair (75.0% decrease) after 30 minutes, and 102.9 TCID50/10Lair (93.9% decrease) 60 minutes after the start of the test. Table 1 shows the influenza virus test results (TCID50/10Lair).

This test was conducted to confirm the effectiveness of test equipment against airborne influenza viruses in a 25m3 space. Test results showed a reduction in airborne influenza virus infectivity of 59.3% in 6 minutes, 75.0% in 30 minutes, and 93.9% in 60 minutes. Therefore, according to the antibacterial method of the present invention, it has been found that by spraying the antibacterial water composition for 60 minutes or more, the antiviral activity value becomes 1.2 or more. (Antiviral activity value = log (control group/infectious titer after time) - log (test group/infectious titer after time))

According to the present invention, since it has antibacterial properties without adding preservatives or bactericidal agents, it has high industrial utility value in a wide range of fields.

Claims (6)

An antibacterial water composition containing at least one amino acid selected from L-cysteine, L-arginine, L-lysine, L-histidine, or salts thereof, wherein the antibacterial water composition has a pH of 10.9. a step of spraying the above antibacterial water composition indoors , generating a mist by using a piezoelectric vibrator or a two-fluid nozzle in the spraying, and suppressing indoor floating viruses; An indoor antibacterial method characterized by comprising the steps of: , antibacterializing the room . The method according to claim 1, wherein in the spraying step, the average particle diameter of the sprayed mist is 10 μm or less. 3. The method according to claim 1 , wherein the antibacterial water composition further contains reducing ion water. According to any one of claims 1 to 3 , the content of the amino acid is in the range of 1.0% by mass to 0.00001% by mass with respect to the total amount of the antibacterial water composition. Method described. The method according to claim 3, wherein the content of the reducing ion water is in the range of 10% by mass to 90% by mass with respect to the total amount of the antibacterial water composition. The method according to any one of claims 1 to 5 , wherein the antibacterial water composition does not contain a pH adjuster.