WO2021075425A1

WO2021075425A1 - Disinfectant including ultrafine-bubble-containing solution

Info

Publication number: WO2021075425A1
Application number: PCT/JP2020/038631
Authority: WO
Inventors: 藤田　豊久; 蘭因張; ジョルジドドビバ; 裕史松井; 宏美黒川; 税鈴木
Original assignee: ベルパック株式会社; ベルテクノサービス株式会社
Priority date: 2019-10-18
Filing date: 2020-10-13
Publication date: 2021-04-22
Also published as: JPWO2021075425A1; WO2021075043A1; JP7450849B2; WO2021075332A1; JPWO2021075332A1; JPWO2021075043A1

Abstract

[Problem] Ultrafine bubbles can be present for a long period of time, and therefore the ROS elimination effect thereof could also persist for a long period of time. The effect of ultrafine bubble beverages containing various gases on reactive oxygen is unknown, and the present invention addresses the study of this effect. [Solution] With this disinfectant containing an ultrafine-bubble-containing organic solution, it is possible to provide a very safe disinfectant having a reactive-oxygen-reinforcing effect and a disinfectant effect, the disinfectant including carbon dioxide ultrafine bubbles and hydrogen ultrafine bubbles and not including chemical substances.

Description

Fungicide containing ultrafine bubble-containing solution

The present invention relates to a disinfectant containing an ultrafine bubble-containing solution, and more particularly to a disinfectant containing an ultrafine bubble-containing solution containing carbon dioxide and hydrogen.

Active oxygen (hereinafter, also referred to as "ROS") is a causative agent of many diseases, while active oxygen has a high reaction activity, so it has a defense function to eliminate foreign substances (microorganisms) that have entered from the outside. I've also come to understand that it has.

When neutrophils such as white blood cells and macrophages recognize foreign substances and toxic substances in the body and take them in and decompose them, active oxygen works to decompose bacteria and the like. White blood cells (neutrophils) phagocytose when bacteria invade the body, and white blood cells use NAD (P) H oxidase to react NADH (NADPH) with H + and oxygen to generate hydrogen peroxide. It is said to have a biological defense mechanism that sterilizes bacteria that are produced and that are still proliferating even after being phagocytosed and protect them from infection.

However, hydroxyl radicals (OH ·) and the superoxide anion radical (· O 2 _^-) active oxygen, such as, for high low stability reactive, difficult to expect a sustained action for such the sterilizing action Is.

On the other hand, Ultrafine Bubble (hereinafter, also referred to as "UFB (Ultrafine Bubble)") has a high negative zeta potential in a neutral solution and has an antioxidant ability. The size of UFB decreases with the passage of time, and it is considered that the UFB can exist even after the passage of one month.

Sufficient studies have not yet been conducted on the sustained antioxidant and bactericidal effects of ultrafine bubbles. For example, hydrogen has a high reducing power, but the antioxidant capacity of nanofine bab hydrogen water (ultrafine bubble aqueous solution containing hydrogen) has not been sufficiently investigated.

Conventionally, there are very few reports on the relationship between ultrafine bubbles and bactericidal action. A disinfectant containing a weakly alkaline stable sodium hypochlorite solution and containing ultrafine bubbles is disclosed (Patent Document 1 below). However, this is based on the bactericidal action of sodium hypochlorite and is not related to the bactericidal action of the ultrafine bubble itself.

Regarding the creation of ultrafine bubbles, Ahmed et al. Considered ultrafine bubble-containing aqueous solutions of air, nitrogen, and oxygen using cylindrical ceramic nanofiltration membranes instead of the conventional method of generation by hydrodynamics, acoustics, particles, and optical cavitation. However, it has been reported that the size of air bubbles can be reduced to the nano level (see Non-Patent Document 1 below).

JP-A-2018-090547

As mentioned above, since the ultrafine bubble can exist for a long period of time, there is a possibility that the peculiar action of the ultrafine bubble such as antioxidant action will continue for a long period of time. However, the effect of ultrafine bubbles of various gases on active oxygen is unknown, and its examination has been an issue.

The present inventors have investigated the antioxidative ability and the effect on various diseases of nanofine bubbles of various gases, and the present inventors include at least one gas of carbon dioxide and hydrogen inside the ultrafine bubbles. We found that the solution containing ultrafine bubbles has a continuous ROS scavenging ability, a cytotoxic effect on cancer cells, and an antitumor effect on cancer-bearing mice, and has an active oxygen scavenging ability, and cytotoxicity on cancer cells. Provided are a solution containing ultrafine bubbles containing at least one gas of carbon dioxide and hydrogen inside the ultrafine bubbles, which has an effect and an antitumor effect on cancer-bearing mice, and a beverage containing the same, and a medicine. It is disclosed in PCT / JP2019 / 041060 that it can be done.

In view of the above problems, the present inventors are studying the antioxidative ability of nanofine bubbles of various gases and their application to various uses. Surprisingly, carbon dioxide ultrafine bubbles and hydrogen ultra It has been found that a fine bubble-containing solution, particularly a mixed solution of an organic solvent / water containing the ultrafine bubble, has an ROS enhancing action and a bactericidal action based on the ROS enhancing action.

Therefore, an object of the present invention is to provide an ultrafine bubble-containing bactericidal agent containing carbon dioxide ultrafine bubbles and hydrogen ultrafine bubbles, which have active oxygen enhancing action and bactericidal action.

That is, the present invention is as follows.
[1]
A disinfectant for organic solvent-based solutions containing hydrogen ultrafine bubbles and carbon dioxide ultrafine bubbles.
[2]
The bactericidal agent according to [1], wherein the organic solvent-based solution is a mixed solution of an organic solvent and water.
[3]
The bactericidal agent according to [2], wherein the organic solvent is an alcohol-based organic solvent.
[4]
The fungicide according to [3], wherein the alcohol-based organic solvent is at least one selected from the group consisting of ethanol, ethylene glycol, and isopropanol.
[5]
The bactericidal agent according to [3] or [4], wherein the alcohol-based organic solvent is ethylene glycol.
[6]
The fungicide according to [5], wherein the ethylene glycol content is 25% to 55%.
[7]
The bactericidal agent according to [1], wherein the organic solvent-based solution comprises one or a plurality of organic solvents.
[8]
The fungicide according to [7], wherein the plurality of organic solvents are isopropanol and kerosene.
[9]
The fungicide according to [8], wherein the isopropanol content is 25% to 35%.
[10]
The bactericidal agent according to any one of [1] to [9], which is prepared by further blowing hydrogen into an aqueous solution containing carbon dioxide ultrafine bubbles to generate hydrogen ultrafine bubbles.
[11]
A method for producing a disinfectant for an organic solvent-based solution containing hydrogen ultrafine bubbles and carbon dioxide ultrafine bubbles. Hydrogen ultrafine is further blown into an ultrafine bubble-containing solution containing carbon dioxide ultrafine bubbles. A method of producing bubbles to produce the disinfectant.

According to the present invention, it is possible to provide an ultrafine bubble-containing bactericidal agent containing carbon dioxide ultrafine bubbles and hydrogen ultrafine bubbles, which have active oxygen enhancing action and bactericidal action.

It is a figure which shows the carbon dioxide ultrafine bubble making apparatus. It is a figure which shows the hydrogen ultra fine bubble making apparatus. It is a figure which shows the main part of the IFA measuring apparatus of the ultrafine bubble which concerns on this invention. It is a figure which shows the stability of a hydrogen ultrafine bubble. It is a figure which shows the adduct of G-CYPMPO and active oxygen in the active oxygen detection method (spin trap method / ESR method) in this invention. It is a figure which shows the hydroxyl radical scavenging action of hydrogen ultrafine bubble and carbon dioxide ultrafine bubble in ethanol (EtOH) aqueous solution. It is a figure which shows the hydroxyl radical scavenging action of hydrogen and carbon dioxide ultrafine bubble in an aqueous solution of ethanol (EtOH). It is a figure which shows the superoxide anion scavenging action of hydrogen ultrafine bubble and carbon dioxide ultrafine bubble in ethanol (EtOH) aqueous solution. It is a figure which shows the superoxide anion scavenging action of hydrogen and carbon dioxide ultrafine bubble in an aqueous solution of ethanol (EtOH). It is a figure which shows the hydroxyl radical scavenging action of hydrogen ultrafine bubble and carbon dioxide ultrafine bubble in an ethylene glycol (EG) aqueous solution. It is a figure which shows the hydroxyl radical scavenging action of hydrogen and carbon dioxide ultrafine bubble in the ethylene glycol (EG) aqueous solution. It is a figure which shows the superoxide anion scavenging action of hydrogen ultrafine bubble and carbon dioxide ultrafine bubble in an ethylene glycol (EG) aqueous solution. It is a figure which shows the superoxide anion scavenging action of hydrogen and carbon dioxide ultrafine bubbles in an aqueous ethylene glycol (EG) solution. It is a figure which shows the bactericidal action of the ultrafine bubble containing bactericidal agent which concerns on this invention.

Hereinafter, embodiments of the present invention will be described in detail.

(Organic solvent solution containing ultrafine bubbles)
The ultrafine bubble-containing fungicide according to the present invention is an organic solvent solution containing ultrafine bubbles. The organic solvent solution containing the ultrafine bubble is an organic solvent solution containing an ultrafine bubble containing both carbon dioxide and hydrogen gases inside the ultrafine bubble.

In the organic solvent solution containing the ultrafine bubbles, at least one gas of carbon dioxide and hydrogen may be contained inside each ultrafine bubble contained in the solution.

The organic solvent solution is a solution containing an organic solvent, and may be a mixed solution of an organic solvent and water, or a solution composed of one or a plurality of organic solvents.

A bubble is a closed space composed of a gas surrounded by a gas other than a gas, and a bubble completely surrounded by a liquid is a planktonic gas. Among these planktonic gases, bubbles having a diameter of 1 micrometer or less are called ultrafine bubbles (Fine Bubble Society Association).

Since ultrafine bubbles are extremely small bubbles, it is known that the bubbles have a negative potential called the zeta potential, and the bubbles can exist in the liquid for an extremely long time.

Hereinafter, carbon dioxide, hydrogen, and an ultrafine bubble-containing solution containing both carbon dioxide and hydrogen are provided inside the ultrafine bubble, respectively, carbon dioxide ultrafine bubble, hydrogen ultrafine bubble, and carbon dioxide and hydrogen ultra. It is called fine bubble.

It is preferable that the ultrafine bubble also has a specific 50% average particle size. Since the gas contained inside the ultrafine bubble has a particle size smaller than a specific 50% average particle size, the stability of the ultrafine bubble is improved and the ultrafine bubble can exist as an ultrafine bubble for a long time. The 50% average particle size in the present invention usually refers to the 50% average particle size with respect to the number of ultrafine bubbles.

The 50% average particle size of the carbon dioxide ultrafine bubble is in the range of 50 nm to 300 nm in water. A 50% average particle size in this range allows it to exist as an ultrafine bubble for several days. It is difficult to prepare ultrafine bubble carbon dioxide with a 50% average particle size less than 50 nm.

The 50% average particle size of the hydrogen ultrafine bubble is in the range of 10 nm to 500 nm in water. A 50% average particle size in this range allows it to exist as an ultrafine bubble for several days. It is difficult to prepare ultrafine bubbles with a 50% average particle size less than 10 nm.

Table 1 shows an example of the particle sizes of carbon dioxide ultrafine bubbles and hydrogen ultrafine bubbles in water, an ethanol (EtOH) aqueous solution, and an ethylene glycol (EG) aqueous solution. Compared with water, in a mixed solution of an organic solvent and water (hereinafter, also referred to as "organic solvent aqueous solution"), all ultrafine bubbles tend to have a larger particle size.

The ultrafine bubble has a specific ultrafine bubble content. In order for the ultrafine bubble-containing solution to exhibit its antioxidant capacity, it is necessary for the ultrafine bubble to contain a certain amount of gas.

The content of the carbon dioxide ultrafine bubbles is preferably in the range of 100 million / mL to 10 billion / mL, and more preferably in the range of 1 billion / mL to 10 billion / mL.

The content of the hydrogen ultrafine bubbles is preferably in the range of 100 million / mL to 100 billion / mL, and further preferably in the range of 100 million / mL to 50 billion / mL. preferable.

(Preparation of ultrafine bubble-containing solution)
Examples of the method for preparing the ultrafine bubble-containing solution include a spiral flow system method, a pressurized dissolution system, an ultrasonic wave method, passage through porous ceramics, and porosity. Various methods such as passing through a plastic film and a double bottle hydrogen generator are known.

The carbon dioxide ultrafine bubble can be prepared by various methods. Among them, the carbon dioxide ultrafine bubble is prepared by passing carbon dioxide from a pressurized tank through porous ceramics (the porous ceramics passing method) or passing through a porous plastic membrane, and then blowing it into a liquid. (See FIG. 1). As the porous plastic film, a film that removes ordinary fine particles can also be used for nanobubble preparation.

The hydrogen ultrafine bubble can be prepared by various methods. Among them, the hydrogen ultra fine bubble is prepared by electrolysis (electrolysis method for producing fine bubble) using a double bottle hydrogen generator (Woo Co., Ltd., Gas & Water Double Hydrogen Bottle (registered trademark)). Fine bubbles are preferred. A solution containing hydrogen ultrafine bubbles can be prepared by the double bottle hydrogen generator (see FIG. 2).

The carbon dioxide and hydrogen ultrafine bubble solution allows carbon dioxide to pass through porous ceramics (the porous ceramics passing method) or a porous plastic film, and then blows into the solution containing hydrogen ultrafine bubbles. It can be prepared by or by causing the carbon dioxide ultrafine bubble to generate hydrogen ultrafine bubble by a double bottle hydrogen generator.

Further, the carbon dioxide and hydrogen ultrafine bubble-containing solution can be prepared by at least one method selected from the group consisting of the pressurized dissolution system and the spiral flow system. According to these methods, a large amount of carbon dioxide and hydrogen ultrafine bubble-containing solution can be prepared.

(Characteristic measurement method for ultrafine bubble-containing solution)
The characteristics of ultrafine bubbles are measured by the Resonant Mass Method, Particle Trajectory Method, Laser Diffraction Method, and Dynamic Light Scattering Method (DLS Method). ), Interactive Force MFP Method (IFA Method), etc. are known.

The characteristics of the ultrafine bubble in the present invention can be measured by various methods. Among them, in the measurement of the ultrafine bubble of the present invention, it is possible to measure an ultrafine bubble having a small particle size equivalent to that of the dynamic light scattering method (DLS) method, and since it has high accuracy, the following interactions occur. It is preferable to use a force device (IFA) measurement method.

The particle size distribution of the ultrafine bubble in the present invention was measured in a solution using the IFA measuring device shown in FIG. 3 (see FIG. 4). According to this measuring device, "the particle size distribution of fine bubbles from several nm to several 100 μm can be measured with high accuracy, and the particle concentration, the light transmittance of the liquid, and the refractive index are not affected. "Measurable" (Patent No. 650265, p. 3, paragraph [0009]). An example of this measuring device is shown in FIG. The measured values were in good agreement with those measured by the Dynamic Light Scattering (DLS) method.

The method for measuring an interaction force device is disclosed in Japanese Patent Application Laid-Open No. 2016-109453 (Invention title "Fine bubble particle size distribution measuring method and measuring device", Japanese Patent No. 6502657).

(Characteristics of ultrafine bubble-containing solution)
The particle diameter of the particle size the ultra-fine bubbles of ultra-fine bubbles was measured by the interaction force system method. The particle size of the carbon dioxide ultrafine bubble (50% average diameter of the bubble size) was measured to be 115 nm in water, and the particle size of the hydrogenated ultrafine bubble (50% average diameter of the bubble size) was 130 nm in water. Measured (see Table 1).

Stability of Ultra Fine Bubbles The change in particle size of the Ultra Fine Bubbles over time was measured by the above-mentioned interaction force device method, and its stability (sustainability) was evaluated. The carbon dioxide ultrafine bubbles existed in distilled water for several days and disappeared after 4 days. The stability of the carbon dioxide ultrafine bubble in distilled water was several days, and the stability due to the ultrafine bubble was confirmed. It was found that the hydrogen ultrafine bubble had a particle size of 25 nm after the lapse of 40 days, and the particle size became smaller with the lapse of time. It was found that the hydrogen ultrafine bubble was stably present even after 40 days (see FIG. 4).

The contents of carbon dioxide ultrafine bubbles and hydrogen ultrafine bubbles in the ultrafine bubble-containing solution in water are 100 million / mL to 10 billion / mL and 0.1, respectively, according to the particle size and density measurement. It was in the range of 100 billion pieces / mL to 100 billion pieces / mL.

The ultrafine bubble according to the present invention has a sufficiently small particle size and its internal content, and can exist stably for a long period of time.

Hydroxy radical (OH ·) and the superoxide anion radical (· O ₂ ^-) Measurement of, G-CYPMPO as a spin trapping agent (R): 2- (5,5-dimethyl- 2-oxo-2-l5- [1,3,2] dioxaphosphinan-2-yl) -2-methyl-3,4-dihydro-2H-pyrro- line N-oxide {2- (5,5-dimethyl-2-oxo-1,3, Spin-using a 2-dioxaphosphinan-2-yl) -3,4-dihydro-2-methyl-2H-pyrrole N-oxide (chemical formula 1 below) and an ESR spectrometer (JES-TE25X manufactured by JEOL). This was done by recording the ESR spectrum of the trapping adduct.

In a hydrogen peroxide solution (0.1 w / v%) under UV irradiation, that is, an adduct of hydroxyl radical (OH ·) and G-CYPMPO®, and in a hypoxanthine / xanthine oxidase system, that is, superoxide. anion radical (· O ₂ ^-) and the ESR spectrum of the adduct of G-CYPMPO (R) shown in FIG. In the figure, the diamond mark (◆) superoxide anion radical (· O ₂ ^-) peaks of adduct, inverse triangle (▼) indicates a peak of hydroxy radicals (OH ·) adduct. The peak-to-peak intensity of both peaks is used for measuring the amount of active oxygen.

The Kohri's ESR spin trap method, which is a typical method, was used for data analysis. The peak-to-peak intensity of the selected ESR line of the free radical adduct was followed in the presence and absence of antioxidants.

It is conceivable that a wrapping reaction with the following free radicals (R) occurs in the presence of a spin trapping agent (SP) and an antioxidant (AO).
R + SP → R adduct rate constant: tk _sp (1)
R + AO → Product rate constant: k _AO (2)
If I ₀ and I are the ESR peak heights in the presence of ST only and ST + AO, respectively, the amount of product in formula (2) is I _0- I. Therefore, I ₀ / I-1 is calculated to quantify the free radical capture capacity.

To quantify the free radical capture capacity, the oxidant species is mixed with the free radical generating system hours prior to ESR measurements. When I ₀ and I have ESR peak heights in the presence of ST alone and ST + oxidant, respectively, the amount of free radical generation system oxidized to the oxidant species is I _0- I. Therefore, I ₀ / I-1 is calculated to quantify the free radical capture capacity.

(Characteristics of ultrafine bubbles in aqueous ethanol solution)
Hydrogen ultrafine bubble, carbon dioxide ultrafine bubble, hydrogen and carbon dioxide ultrafine bubble (hydrogen blown) in an ethanol aqueous solution (50%, 20%, 10%) which is a form of a mixed solvent of the organic solvent and water. Next, the active oxygen (ROS) scavenging action of carbon dioxide injection: H2 ⇒ CO2, and then hydrogen injection: CO2 ⇒ H2) was examined, and the results are shown in Table 2.

As in water, carbon dioxide ultrafine bubbles show a strong hydroxyl radical scavenging effect (Table 2, 3, 8 and 13), and hydrogen ultrafine bubbles have a hydroxyl radical (HO) scavenging effect of carbon dioxide. It was weaker than the ultrafine bubble (Table 2, 2, 7, 12). On the other hand, regarding the superoxide anion scavenging action, all ultrafine bubbles showed a weak scavenging action at low ethanol concentrations (20%, 10%) (Table 2, 22, 23, 27, 28).

In an ethanol aqueous solution, the hydroxyl radical scavenging action of carbon dioxide and hydrogen ultrafine bubbles is different from that in water, and carbon dioxide blowing (H2⇒CO2) is next to hydrogen blowing and then hydrogen blowing. It was stronger than the inclusion (CO2⇒H2) (4, 9, 14 in Table 2).

Carbon dioxide and hydrogen In the carbon dioxide injection (H2⇒CO2) next to the hydrogen injection of the ultrafine bubble, the hydroxyl radical scavenging action is enhanced by the carbon dioxide injection after the hydrogen injection (Table 2, 4, 9, 14). On the other hand, in the hydrogen injection (CO2⇒H2) after carbon dioxide injection, hydroxyl radicals are eliminated by carbon dioxide injection, but hydroxyl radicals are regenerated by hydrogen injection, and the hydroxyl radical scavenging action is reduced ( Table 2, 5, 10, 15).

That is, in carbon dioxide and hydrogen ultrafine bubbles (CO2⇒H2), the enhancement and persistence of hydroxyl radical activity was observed due to the re-elevation of hydroxyl radicals.

A re-elevation of the superoxide anion of carbon dioxide and hydrogen ultrafine bubbles was also observed, but the effect was weakly observed only in the low-concentration ethanol aqueous solution (10%, 20%) (Table 2, 25, 30). ..

From the above, in an aqueous ethanol solution, carbon dioxide and hydrogen ultrafine bubbles (CO2⇒H2) showed a hydroxyl radical enhancing effect (

Experiments

5, 10 and 15 in Table 2), and a weak superoxide anion enhancing effect was also observed. (25, 30 in Table 2). From this point, the aqueous ethanol solution containing ultrafine bubbles can be mentioned as one of the fungicides according to the present invention.

(Characteristics of ultrafine bubbles in other organic solvent systems)
The organic solvent-based disinfectant according to the present invention preferably contains the disinfectant as a mixed solvent of an organic solvent and water. Further, the organic solvent is preferably an alcohol-based organic solvent. The ethanol aqueous solution is a form of a mixed solution of the alcohol-based organic solvent and water. Examples of the alcohol-based organic solvent include ethylene glycol and isopropanol in addition to ethanol because ROS in the solution is enhanced.

The organic solvent system in the present invention is not particularly limited as long as carbon dioxide ultrafine bubbles and hydrogen ultrafine bubbles can exist and ROS in the solution is enhanced.

Similarly, as the mixed solution of the alcohol-based organic solvent and water, carbon dioxide ultrafine bubbles and hydrogen ultrafine bubbles can be present in the solution, as long as the ROS in the solution is enhanced. There is no particular limitation. In the alcohol-based organic solvent, it is considered that hydroxyl radical (HO ·) is further generated by ROS.

Ethylene glycol is preferable as the alcohol-based organic solvent. In this case, the ethylene glycol content in the mixed solvent of ethylene glycol and water is preferably 25% to 55% from the viewpoint of regeneration of active oxygen, particularly hydroxyl radical.

Examples of the organic solvent-based solution in the present invention include a mixed solvent of isopropanol and kerosene. This solution does not contain water, but ultrafine bubbles can be present. In the mixed solvent, the isopropanol content is preferably 25% to 55% from the viewpoint of regeneration of active oxygen, particularly hydroxyl radical.

For the regeneration of active oxygen, especially hydroxyl radical, in the organic solvent system, the following mechanism of Chemical formula 2 can be assumed.

Hydroxyl radicals are generated by light in the presence of hydrogen peroxide H2O2 in alcohol-based organic solvents such as ethanol and ethylene glycol. By blowing carbon dioxide into a solution, hydrogen peroxide H2O2 is reduced by the reactions of (2), (3) and (4) of Chemical formula 2. Further, by blowing hydrogen into the solution, the hydroxyl radical generated by light by the reaction (1) and the remaining hydrogen peroxide H2O2 generate a superoxide anion by the reactions (7) and (8). Here, when an alcohol-based organic solvent such as ethanol or ethylene glycol and a superoxide anion coexist, more hydrogen peroxide is generated by the reactions (12) and (13) by blowing hydrogen. A mechanism is conceivable.

(Fungicide containing ultrafine bubbles)
The disinfectant according to the present invention is characterized by being a disinfectant as the above-mentioned organic solvent-based solution containing hydrogen ultrafine bubbles and carbon dioxide ultrafine bubbles. Active oxygen regenerated in an organic solvent system containing the hydrogen ultrafine bubbles and carbon dioxide ultrafine bubbles, particularly hydroxyl radicals, is expected to have antibacterial, sterilizing, and bactericidal effects on bacteria and microorganisms.

Furthermore, since it is not necessary to contain a bactericidal chemical substance such as hypochlorous acid, the bactericidal agent according to the present invention is preferable from the viewpoint of high safety and low environmental load.

The disinfectant according to the present invention is an organic solvent-based disinfectant containing hydrogen ultrafine bubbles and carbon dioxide ultrafine bubbles, and is further prepared by hydrogen injection (CO2⇒H2) after carbon dioxide injection. It is unique in that.

(Method for producing a fungicide according to the present invention)
The method for producing a bactericide according to the present invention is a method for producing an ultrafine bubble-containing bactericide containing hydrogen ultrafine bubbles and carbon dioxide ultrafine bubbles, and is an ultrafine bubble-containing organic solvent containing carbon dioxide ultrafine bubbles. This is a method of producing an organic solvent containing hydrogen ultrafine bubbles and carbon dioxide ultrafine bubbles by further injecting hydrogen into the water.

In the bactericidal agent according to the present invention, active oxygen regenerated in an organic solvent system containing the hydrogen ultrafine bubble and carbon dioxide ultrafine bubble, particularly hydroxyl radical, has an antibacterial action and a sterilizing action against bacteria, microorganisms and the like. Expected to have bactericidal action. This bactericidal action was confirmed by the bactericidal action against Escherichia coli in the following examples (Fig. 14).

Hereinafter, the present invention will be described in detail with reference to examples, but the present invention is not limited to these examples.

(1) Preparation of Ultrafine Bubble-Containing Aqueous Solution Experimental Example 1: Preparation of Ultrafine Bubble Carbon Dioxide Containing Solution The carbon dioxide ultrafine bubble-containing solution (aqueous solution) is prepared using a magnetic stirrer as shown in FIG. _{With stirring, carbon dioxide (CO 2} , 99.5%) in argon gas from a pressurized tank passes through porous ceramics in a beaker containing 200 mL of water at a rate of 0.4 L / min for 30 minutes. (The above-mentioned porous ceramics passing method) was prepared (hereinafter, also referred to as _{"containing solution CO 2").} The ultrafine bubble carbon dioxide in the contained solution is also hereinafter referred to as "UFB / CO ₂ ".

Experimental Example 2: Preparation of hydrogen ultrafine bubble-containing solution The hydrogen ultrafine bubble-containing solution (aqueous solution) is subjected to a double bottle hydrogen generator (manufactured by Woo, Gas & Water Double Hydrogen Bottle) as shown in FIG. It was produced by electrolysis using (trademark)) (hereinafter, also referred to as _{“UFB / EH 2”).} The hydrogen ultrafine bubble-containing solution (aqueous solution) was prepared in 200 mL of water for 30 minutes (hereinafter, also referred to as _{“containing solution EH 2”).} The concentration of ultrafine bubble hydrogen by electrolysis was 0.15% by volume, and was quantified using a specific gravity bottle.

Experimental Example 3: Preparation of carbon dioxide and hydrogen ultrafine bubbles according to the present invention The ultrafine bubble carbon dioxide and hydrogen mixed solution is added to the solution containing ultrafine bubble hydrogen (prepared in the previous item (2)) in the previous item (1). _{), While stirring using a magnetic stirrer, blow carbon dioxide (CO 2} , 99.5%) in argon gas from the pressurized tank for 30 minutes at a rate of 0.4 L / min. (Hereinafter, also referred to as "mixed solution CO ₂ / EH _2"). Alternatively, the ultrafine bubble carbon dioxide and hydrogen mixed solution is electrolyzed into the carbon dioxide ultrafine bubble-containing solution (prepared in the previous item (1)) for 30 minutes according to the method in the previous item (2). (Hereinafter, also referred to as "mixed solution EH ₂ / CO _2").

(2) Measurement of Ultra Fine Bubble by IFA Measuring Device The ultra fine bubble is measured in water using the IFA measuring device of FIG. 3, according to JP-A-2016-109453, "Fine Bubble Particle Size Distribution Measuring Method and Measuring Device". The particle size distribution was measured by the indicated IFA method (see FIG. 4).

(2) ROS scavenging action of ultrafine bubble-containing aqueous solution Experimental example 4: Generation of active oxygen (ROS) Hydroxyl radical (OH ·) is 0 under 5s ultraviolet radiation of a UV radiator (made by UV LIGHTSOURCE, SUPERCURE-203S). It was produced by decomposition of 1 w / t% hydrogen peroxide (H ₂ O _2). A mixture of 90 μL of 0.2 w / t% hydrogen peroxide and 20 μL of 100 mM G-CYPMPO was transferred to a disposable borosilicate ESR cell, and the ESR spectrum of the adduct of hydroxyl radical and G-CYPMPO was analyzed.

Superoxide anion radical (· _{O 2} ^-) it is, was generated in hypoxanthine / xanthine (HX / XO) system. A mixture of 10.970 units / mL XO, 20 mM HX 20 μL, and 100 mM G-CYPMPO 20 μL was transferred to a disposable borosilicate ESR cell and the ESR spectrum of the adduct of hydroxyl radicals and G-CYPMPO was analyzed.

Experimental Example 5: Measurement of active oxygen by ESR A novel radical trapper 2- (5,5-dimethyl-2-oxo-2-l5-[1,3,2] dioxaphosphinan-2-yl) -2- methyl-3,4-dihydro-2H-pyrro-line N-oxide {2-(5,5-dimethyl-2-oxo-1,3,2-dioxaphosphinan-2-yl) -3,4-dihydro-2 -Methyl-2H-pyrrole N-oxide, G-CYPMPO® was used to capture the free radicals of reactive oxygen species. 25 mg of G-CYPMPO® (100 mL) was dissolved in 2 mL of ultrapure water.

The ESR spectrum of the spin trapping adduct was recorded using an ESR spectrometer (JES-TE25X manufactured by JEOL). Typical ESR measurement conditions were as follows.
Microwave power: 4 mW, microwave frequency: 9.2 GHz, magnetic field: 328.0 mT, field sweep with: ± 7.5 mT, field modulation: 0.16 mT, sweep time: 1 minute, 0.003663 mT / Points, all 4096 points, ESR measurements were performed at room temperature.

In the presence of the spin trapping agent (SP) and the antioxidant (AO), a wrapping reaction with the following free radicals (R) occurs.
R + SP → R adduct rate constant: tk _sp (1)
R + AO → Product rate constant: k _AO (2)
If I ₀ and I are the ESR peak heights in the presence of ST only and ST + AO, respectively, the amount of product in formula (2) is I _0- I. Therefore, I ₀ / I-1 was calculated to quantify the free radical capture capacity.

To quantify the free radical capture capacity, the oxidant species is mixed with the free radical generating system hours prior to ESR measurement. When I ₀ and I have ESR peak heights in the presence of ST alone and ST + oxidant, respectively, the amount of free radical generation system oxidized to the oxidant species is I _0- I. Therefore, I ₀ / I-1 was calculated to quantify the free radical capture capacity.

Examples 1 to 5: Intensity of hydroxyl radical (OH ·) of the ultrafine bubble-containing ethylene glycol (EG) solution (50%) according to the present invention Control Example 1: Ultrafine bubble-free, Reference Example 2: Hydrogen Ultra Fine bubble containing, Reference Example 3: Carbon dioxide ultrafine bubble containing, Reference Example 4: Carbon dioxide and hydrogen ultrafine bubble containing (prepared by blowing hydrogen to carbon dioxide in order), and Example 5: Carbon dioxide and hydrogen ultrafine The hydroxyl radical (OH ·) strength of each 50% ethylene glycol solution containing bubbles (prepared by blowing in the order of carbon dioxide to hydrogen) was examined by ESR measurement. _{The I 0} / I-1 obtained from the ESR spectrum of the G-CYPMPO® adduct of the ultrafine bubble-containing solution is shown in FIGS. 10 and 11.

Based on FIGS. 10 and 11, the intensities of hydroxyl radicals are shown in Table 3 with strong ⊚, moderate ◯, weak Δ, and x with little or no recognition. Example 5: Carbon dioxide and hydrogen ultrafine bubble containing (prepared by blowing in the order of carbon dioxide to hydrogen) is compared with Reference Example 4: containing carbon dioxide and hydrogen ultrafine bubbles (prepared by blowing in the order of hydrogen to carbon dioxide). Therefore, stronger hydroxyl radicals (Example 5: ○, Reference Example 4: ×) were observed.

Examples 6 to 10: Strength of hydroxyl radical (OH ·) of ultrafine bubble-containing ethylene glycol aqueous solution (30%) according to the present invention Control example 6: Ultrafine bubble free, Reference example 7: Hydrogen ultrafine bubble content , Reference Example 8: Carbon dioxide ultrafine bubble containing, Reference Example 9: Carbon dioxide and hydrogen ultrafine bubble containing (prepared by blowing hydrogen to carbon dioxide in order), and Example 10: Carbon dioxide and hydrogen ultrafine bubble containing ( The hydroxyl radical (OH ·) strength of each 30% ethylene glycol aqueous solution (prepared by blowing in the order of carbon dioxide to hydrogen) was examined by ESR measurement. _{The I 0} / I-1 obtained from the ESR spectrum of the G-CYPMPO® adduct of the ultrafine bubble-containing solution is shown in FIGS. 10 and 11.

Based on FIGS. 10 and 11, the intensities of hydroxyl radicals are shown in Table 3 with strong ⊚, moderate ◯, weak Δ, and x with little or no recognition. Example 10: Carbon dioxide and hydrogen ultrafine bubble containing (prepared by blowing in the order of carbon dioxide to hydrogen) is compared with Reference Example 9: containing carbon dioxide and hydrogen ultrafine bubbles (prepared by blowing in the order of hydrogen to carbon dioxide). Therefore, stronger hydroxyl radicals (Example 10: ⊚, Reference Example 9: ×) were observed.

Examples 11 to 15: Strength of super oxide anion (O2 ·-) of ultrafine bubble-containing ethylene glycol aqueous solution (50%) according to the present invention Control example 11: Ultrafine bubble-free, Reference example 12: Hydrogen Ultrafine Bubble containing, Reference Example 13: Carbon dioxide ultrafine bubble containing, Reference Example 14: Carbon dioxide and hydrogen ultrafine bubble containing (prepared by blowing in the order of hydrogen to hydrogen), and Example 15: Carbon dioxide and hydrogen ultrafine bubble The superoxide anion (O2-) scavenging action of each 50% ethylene glycol aqueous solution of the content (prepared by blowing in the order of carbon dioxide to hydrogen) was examined by ESR measurement. _{The I 0} / I-1 obtained from the ESR spectrum of the G-CYPMPO® adduct of the ultrafine bubble-containing solution is shown in FIGS. 12 and 13.

Based on FIGS. 12 and 13, the intensities of superoxide anions are shown in Table 3 with strong ⊚, moderate ◯, weak Δ, and little or no x. Example 15: Carbon dioxide and hydrogen ultrafine bubble containing (prepared by blowing in the order of carbon dioxide to hydrogen) is compared with Reference Example 14: containing carbon dioxide and hydrogen ultrafine bubbles (prepared by blowing in the order of hydrogen to carbon dioxide). The latter was slightly stronger in the superoxide anion (Example 15: ○, Reference example 14: ⊚).

Examples 16 to 20: Strength of super oxide anion (O2 ·-) of ultrafine bubble-containing ethylene glycol aqueous solution (30%) according to the present invention Control example 16: Ultrafine bubble-free, Reference example 17: Hydrogen Ultrafine Bubble containing, Reference Example 18: Carbon dioxide ultrafine bubble containing, Reference Example 19: Carbon dioxide and hydrogen ultrafine bubble containing (prepared by blowing in the order of hydrogen to carbon dioxide), and Example 20: Carbon dioxide and hydrogen ultrafine bubble The strength of the superoxide anion (O2 ·-) of each 30% ethylene glycol aqueous solution containing (prepared by blowing in the order of carbon dioxide to hydrogen) was examined by ESR measurement. _{The I 0} / I-1 obtained from the ESR spectrum of the G-CYPMPO® adduct of the ultrafine bubble-containing solution is shown in FIGS. 12 and 13.

Based on FIGS. 12 and 13, the intensities of superoxide anions are shown in Table 3 with strong ⊚, moderate ◯, weak Δ, and x with little or no scavenging effect. Reference Example 20: Carbon dioxide and hydrogen ultrafine bubble containing (prepared by blowing in order from carbon dioxide to hydrogen) and Reference Example 19: Carbon dioxide and hydrogen ultrafine bubble containing (prepared by blowing in order from hydrogen to carbon dioxide) The latter was slightly stronger in the superoxide anion (Example 20: ○, Reference example 19: ⊚).

As shown in FIGS. 10 to 13 and Table 3, regarding the hydroxyl radical intensity of the ultrafine bubble-containing ethylene glycol aqueous solution according to the present invention, carbon dioxide and hydrogen ultrafine bubbles (prepared by blowing carbon dioxide to hydrogen in this order) are used. It turned out to be preferable to use.

Examples 1 to 5: Bactericidal effect of the ultrafine bubble-containing ethylene glycol (EG) solution (50%) according to the present invention Control Example 1: No additive, Control Example 2: Ultrafine bubble-free, Reference Example 4: 50% ethylene each containing carbon dioxide and hydrogen ultrafine bubbles (prepared by blowing in the order of carbon dioxide to carbon dioxide), and Example 5: containing carbon dioxide and hydrogen ultrafine bubbles (prepared by blowing in the order of carbon dioxide to hydrogen) The bactericidal effect of the glycol solution was evaluated by observing survival after 48 hours of incubation using Escherichia coli (Fig. 14).

In Example 5 (FIG. 14C), the survival of Escherichia coli was significantly suppressed as compared with Control Example 1 (FIG. 14D), as in Control Example 2 (FIG. 14A). In addition, the survival of Escherichia coli was further suppressed as compared with Reference Example 4 (FIG. 14a) (Table 3: Example 5). From the above, the bactericidal effect of the regenerated hydroxyl radical of the ultrafine bubble-containing bactericidal agent according to the present invention was confirmed.

Although the present invention has been described above using the embodiments and examples, it goes without saying that the technical scope of the present invention is not limited to the scope described in the above embodiments. It will be apparent to those skilled in the art that various changes or improvements can be made to the above embodiments. Further, it is clear from the description of the scope of claims that the form to which such a modification or improvement is added may be included in the technical scope of the present invention.

310 Electric balance 320 Platinum 330 Sample solution 340 Particle behavior part 341 Hemisphere 342 Flat plate 343 Piezo stage

Claims

A disinfectant for organic solvent-based solutions containing hydrogen ultrafine bubbles and carbon dioxide ultrafine bubbles.
The bactericidal agent according to claim 1, wherein the organic solvent-based solution is a mixed solution of an organic solvent and water.
The bactericidal agent according to claim 2, wherein the organic solvent is an alcohol-based organic solvent.
The bactericidal agent according to claim 3, wherein the alcohol-based organic solvent is at least one selected from the group consisting of ethanol, ethylene glycol, and isopropanol.
The bactericidal agent according to claim 3 or 4, wherein the alcohol-based organic solvent is ethylene glycol.
The bactericidal agent according to claim 5, wherein the ethylene glycol content is 25% to 55%.
The bactericidal agent according to claim 1, wherein the organic solvent-based solution comprises one or a plurality of organic solvents.
The fungicide according to claim 7, wherein the plurality of organic solvents are isopropanol and kerosene.
The bactericidal agent according to claim 8, wherein the content of the isopropanol is 25% to 35%.
The bactericidal agent according to any one of claims 1 to 9, which is prepared by further blowing hydrogen into an aqueous solution containing carbon dioxide ultrafine bubbles to generate hydrogen ultrafine bubbles.
A method for producing a disinfectant for an organic solvent-based solution containing hydrogen ultrafine bubbles and carbon dioxide ultrafine bubbles. Hydrogen ultrafine is further blown into an ultrafine bubble-containing solution containing carbon dioxide ultrafine bubbles. A method of producing bubbles to produce the disinfectant.