JP7641230B2 - Virus inactivation composition, method for enhancing virus inactivation efficacy, and virus inactivation method - Google Patents

Description

The present invention relates to a virus inactivation agent composition comprising a virus inactivating component in an aqueous solution, a method for enhancing virus inactivation efficacy, and a virus inactivation method.

Food poisoning caused by norovirus, a member of the Calicivirus family, has become a problem in food manufacturing and cooking facilities, medical facilities, educational facilities, and the like. Known methods for inactivating norovirus include using a sodium hypochlorite solution and disinfecting with boiling water. However, disinfection with hypochlorite is difficult to use, as it bleaches the treated surface and generates a foul odor, and is not safe enough because it generates chlorine gas when mixed with acidic bleach. In addition, with boiling water disinfection, the temperature drops due to contact with the object to be disinfected, making it difficult to maintain the desired temperature. Furthermore, there is a risk of burns from boiling water. In light of this situation, there is a demand for a virus inactivator that is both safe and highly effective at inactivating viruses.

In order to inactivate such viruses, virus inactivators containing lower alcohols as virus inactivation components are widely used. For example, Patent Document 1 discloses a viricide containing at least 70% by weight of ethanol and/or propanol and 0.5 to 5% by weight of a short-chain organic acid.

Furthermore, Patent Document 2 discloses a method for inactivating non-enveloped virus particles by contacting the virus particles with an alcohol composition containing an effect-enhancing amount of one or more enhancers selected from the group consisting of C1-6 alcohol, and cationic oligomers or polymers, proton donors, chaotropic agents, and mixtures thereof, and wherein when the alcohol composition contains a proton donor, the alcohol composition further contains a synergistic amount of a cationic oligomer or polymer, and also discloses that the alcohol concentration is 50% by mass or more.

Furthermore, Patent Document 3 discloses a disinfectant solution that contains (A) 50 to 70% by weight of ethanol and (B) 0.05 to 4.50% by weight of at least one selected from the group consisting of organic acids, organic acid salts, and ethanolamines, and has a pH of 6 to 12.

Japanese Unexamined Patent Publication No. 63-14702 Special Publication No. 2009-526060 JP 2016-210807 A

However, the virus inactivators in Patent Documents 1 to 3 all have a relatively high alcohol concentration, which causes problems with flammability and alcohol irritation. Therefore, caution is required when using them in places where fire is used, such as around the kitchen. In addition, there is a risk of paint or wax peeling off when a disinfectant containing ethanol is applied to a resin component, and there is also the problem that it is difficult to use for people who do not like the smell of ethanol or whose hands become rough from ethanol. In addition, the pH of Patent Document 3 is neutral to alkaline, ranging from 6 to 12, so there is also the problem of corrosion due to alkali. Furthermore, since food, tableware, etc. are present around the kitchen, there has been a demand for a highly safe virus inactivator that does not affect health even if it adheres to them.

In view of the above problems, the present invention aims to provide a virus inactivator composition that can reduce the amount of ethanol, a virus inactivating ingredient, has a high virus inactivation effect, and is also safe, as well as a method for enhancing virus inactivation efficacy and a virus inactivation method.

In order to achieve the above object, the present invention provides a virus inactivator composition comprising (A) 10% by mass to 60% by mass of ethanol as a virus inactivation component, (B) fumaric acid as a virus inactivation efficacy enhancing component, mixed with (C) water to form an acidic aqueous solution , wherein the blending amount of the virus inactivation efficacy enhancing component is 0.05% by mass or more .

The present invention also relates to a virus inactivator composition comprising (A) 10% by mass to 60% by mass of ethanol as a virus inactivating component, and (B) one or more components selected from phosphoric acid and citric acid as a virus inactivation efficacy enhancing component, mixed with (C) water to form an acidic aqueous solution, wherein the blending amount of the virus inactivation efficacy enhancing component is 0.05% by mass or more and 0.5% by mass or less (however, excluding the cases where citric acid and oyster extract are contained, the cases where a glycerol fatty acid ester and grapefruit seed extract are contained, and the cases where a phloroglucinol polymer is contained) .

The present invention also provides a method for producing an acidic aqueous solution comprising mixing (A) 10% by mass to 60% by mass of ethanol as a virus inactivation component, and (B) one or more components selected from phosphoric acid and citric acid as a virus inactivation effect enhancing component with (C) water,
The virus inactivator composition is characterized in that the blending amount of the virus inactivation efficacy enhancing component is 0.05% by mass or more and 0.5% by mass or less ( however, excluding the cases where the composition contains citric acid and a persimmon extract, where the composition contains a glycerol fatty acid ester and a grapefruit seed extract, where the composition contains a phloroglucinol polymer, where the composition contains a zinc-containing compound, and where the composition contains PEG/PPG-20-35/5-30 dimethicone).

The present invention also provides a method for producing an acidic aqueous solution comprising mixing (A) 10% by mass to 60% by mass of ethanol as a virus inactivation component, and (B) one or more components selected from phosphoric acid and citric acid as a virus inactivation effect enhancing component with (C) water,
The virus inactivator composition is characterized in that the blending amount of the virus inactivation efficacy enhancing component is 0.05% by mass or more and 0.5% by mass or less ( however, excluding the cases where the composition contains citric acid and a persimmon extract, where the composition contains a glycerol fatty acid ester and a grapefruit seed extract, where the composition contains a phloroglucinol polymer, where the composition contains a zinc-containing compound, where the composition contains PEG/PPG-20-35/5-30 dimethicone, and where the composition contains a moisturizer) .

Furthermore, the present invention is characterized in that, in the virus inactivator composition having the above-mentioned configuration, the blending mass ratio (A)/(B) of the virus inactivating component to the virus inactivation effect-enhancing component satisfies 1.30≦ _log10 [(A)/(B)]≦3.10.

The present invention is further characterized in that the virus inactivating agent composition of the above configuration contains 10% to 40% by mass of ethanol as the virus inactivating component.

The present invention is further characterized in that the virus inactivating agent composition of the above configuration contains 30% to 60% by mass of ethanol as the virus inactivating component.

The present invention also relates to a method for enhancing virus inactivation efficacy, comprising adding (B) fumaric acid as a virus inactivation efficacy enhancing component to a virus inactivator composition containing (A) 10% by mass to 60% by mass of ethanol as a virus inactivation component and (C) water, wherein the amount of the virus inactivation efficacy enhancing component added is 0.05% by mass or more relative to the virus inactivator composition .

The present invention also relates to a method for enhancing virus inactivation efficacy, comprising adding (B) one or more components selected from phosphoric acid and citric acid as virus inactivation efficacy enhancing components to a virus inactivator composition containing (A) 10% by mass to 60% by mass of ethanol as a virus inactivating component, and (C) water, in an amount of 0.05% by mass or more and 0.5% by mass or less of the virus inactivating efficacy enhancing components added relative to the virus inactivator composition (however, excluding the cases where citric acid and oyster extract are contained, the cases where a glycerol fatty acid ester and grapefruit seed extract are contained, and the cases where a phloroglucinol polymer is contained) .

The present invention also relates to a method for inactivating non-enveloped viruses, which comprises contacting the virus with the virus inactivator composition having the above-described configuration.

According to the first aspect of the present invention, 10% to 60% by mass of ethanol (A) as a virus inactivating component, and (B) fumaric acid as a virus inactivation efficacy enhancing component are mixed with (C) water to form an acidic aqueous solution, and the amount of the virus inactivation efficacy enhancing component is set to 0.05% by mass or more , thereby providing a virus inactivator composition with a low ethanol concentration. Therefore, the virus inactivator composition is free from the risk of flammability or irritation to the skin caused by ethanol or alkali, has an excellent feel when used, and has high virus inactivation efficacy.

According to the second to fourth aspects of the present invention, a virus inactivator composition having a low ethanol concentration is obtained by mixing (A) 10% to 60% by mass of ethanol as a virus inactivation component, and (B) one or more types selected from phosphoric acid and citric acid as a virus inactivation efficacy enhancing component with (C) water to prepare an acidic aqueous solution, and setting the blending amount of the virus inactivation efficacy enhancing component to 0.05% to 0.5% by mass. Thus, a virus inactivator composition having excellent usability, high virus inactivation efficacy, and no risk of flammability or irritation to the skin caused by ethanol or alkali is obtained.

Furthermore, according to the fifth aspect of the present invention, in the virus inactivator composition of any of the first to fourth aspects, the blending mass ratio (A)/(B) of the virus inactivating component to the virus inactivation efficacy enhancing component satisfies 1.30≦ _log10 [(A)/(B)]≦3.10, so that the blending amount of the virus inactivating efficacy enhancing component relative to the virus inactivating component can be made necessary and sufficient.

Furthermore, according to the sixth aspect of the present invention, in the virus inactivator composition of any one of the first to fifth aspects, the amount of ethanol, which is a virus inactivating component, is set to 10% by mass to 40% by mass, thereby providing a virus inactivator composition with excellent usability in which the odor and irritation of ethanol are further reduced.

Furthermore, according to the seventh aspect of the present invention, in the virus inactivator composition of any one of the first to fifth aspects, by setting the blending amount of ethanol, which is a virus inactivating component, to 30% by mass to 60% by mass, a virus inactivator composition is obtained in which the virus inactivation effect is further enhanced.

Furthermore, according to the eighth aspect of the present invention, by adding (B) fumaric acid as a virus inactivation efficacy enhancing component to a virus inactivator composition containing (A) 10% by mass to 60% by mass of ethanol as a virus inactivating component and (C) water , and setting the amount of the virus inactivation efficacy enhancing component to be added to be 0.05% by mass or more relative to the virus inactivator composition , the virus inactivation efficacy of ethanol, which is a virus inactivating component, can be improved, and a virus inactivator composition that contains a small amount of ethanol and has high virus inactivation power can be produced.

Furthermore, according to the ninth aspect of the present invention, by adding (B) one or more types of virus inactivation effect enhancing components selected from phosphoric acid and citric acid as a virus inactivation effect enhancing component to a virus inactivator composition containing (A) 10% by mass to 60% by mass of ethanol as a virus inactivation component, and (C) water, and setting the amount of the virus inactivation effect enhancing component to be added in an amount of 0.05% by mass or more and 0.5% by mass or less relative to the virus inactivator composition, the virus inactivation effect of ethanol, which is a virus inactivation component, can be improved, and a virus inactivator composition that contains a small amount of ethanol and has high virus inactivation power can be produced.

Furthermore, according to the tenth aspect of the present invention, by contacting a non-enveloped virus with the virus inactivator composition of the first aspect, an effective method for inactivating a non-enveloped virus which has low drug susceptibility and is difficult to inactivate can be achieved.

The virus inactivator composition of the present invention will be described in detail below. The virus inactivator composition of the present invention is prepared by mixing (A) 10% to 60% by mass of ethanol as a virus inactivation component, (B) one or more specific organic acids or inorganic acids as a virus inactivation effect enhancing component, with (C) water to form an acidic aqueous solution.

It was known that ethanol (A), which is blended as a virus inactivating component in the virus inactivator composition of the present invention, has a high virus inactivating effect. However, the present inventors were the first to discover that the combined use of ethanol with a specific organic acid and/or inorganic acid described below exhibits a synergistic virus inactivating effect.

If the amount of ethanol in the virus inactivating composition of the present invention is too small, sufficient virus inactivation effect may not be obtained. On the other hand, if the amount is too large, there are problems such as increased flammability and irritation.

In the virus inactivator composition of the present invention, as shown in the examples described below, ethanol is blended in an amount of 10% by mass or more and 60% by mass or less based on the entire virus inactivator composition in order to obtain sufficient virus inactivation efficacy and reduce flammability and irritation caused by ethanol. In addition, blending ethanol in an amount of 10% by mass or more and 40% by mass or less based on the entire virus inactivator composition is preferable because it can further reduce the odor and irritation of ethanol. In addition, blending ethanol in an amount of 30% by mass or more and 60% by mass or less based on the entire virus inactivator composition is preferable because it can further enhance the virus inactivation effect.

Specific examples of organic acids that are blended into the virus inactivator composition of the present invention as the virus inactivation efficacy enhancing component (B) include fumaric acid and citric acid. Specific examples of inorganic acids include phosphoric acid. These organic acids and/or inorganic acids may be used alone or in combination of two or more.

Among the above organic acids and/or inorganic acids, the use of fumaric acid and phosphoric acid in particular can significantly improve the virus inactivation efficacy of ethanol, a virus inactivation ingredient, at a low blend amount.

The pH of the virus inactivator composition of the present invention is preferably 1 to 6. This makes the virus inactivator composition acidic to weakly acidic, and compared to alkaline virus inactivator compositions, it is not corrosive to the skin and provides a virus inactivator composition with excellent usability.

The amount of the viral inactivation efficacy enhancing component in the viral inactivator composition of the present invention is not particularly limited, but if the amount is too small, a sufficient enhancing effect on the viral inactivation efficacy of ethanol may not be obtained.

As shown in the examples below, in order to obtain a sufficient virus inactivation efficacy enhancing effect, the organic acid and/or inorganic acid, which is a virus inactivation efficacy enhancing component, is preferably blended in an amount of 0.05% by mass or more, more preferably 0.07% by mass or more, and even more preferably 0.1% by mass or more, relative to the entire virus inactivator composition. Also, the organic acid and/or inorganic acid, which is a virus inactivation efficacy enhancing component, is preferably blended in an amount of 5% by mass or less, more preferably 2% by mass or less, and even more preferably 1% by mass or less, relative to the entire virus inactivator composition.

In the present invention, the blending mass ratio (A)/(B) of the virus inactivating component (A) and the virus inactivation efficacy enhancing component (B) preferably satisfies 1.30≦ _log10 [(A)/(B)]≦3.10, more preferably satisfies 1.50≦ _log10 [(A)/(B)]≦3.00, and further preferably satisfies 2.10≦ _log10 [(A)/(B)]≦2.90.

The virus inactivator composition of the present invention can be blended with a surfactant as necessary. For example, surfactants have the property of generating foam (foamability), and by using a surfactant with excellent foamability, dripping when the virus inactivator composition of the present invention is sprayed onto a wall surface with a trigger spray or the like is suppressed and the applied area is also easily visible.

As the surfactant to be incorporated in the virus inactivator composition of the present invention, any of anionic surfactants, cationic surfactants, nonionic surfactants, and amphoteric surfactants can be suitably used. Among these, cationic surfactants that have antiviral activity in addition to surface activity can be more suitably used. The amount of surfactant incorporated in the virus inactivator composition of the present invention is not particularly limited, but is preferably 0.1% by mass or more and 10% by mass or less.

Examples of anionic surfactants include fatty acid soaps, alkylbenzene sulfonates, linear alkylbenzene sulfonates, alkyl sulfates, α-olefin sulfonates, alkyl phosphates, polyoxyethylene alkyl ether sulfates, polyoxyethylene alkyl phenyl ether sulfates, and polyoxyethylene alkyl ether phosphates.

Examples of cationic surfactants include quaternary ammonium salts such as lauryltrimethylammonium chloride, benzalkonium chloride, benzethonium chloride, didecyldimethylammonium chloride, and chlorhexidine gluconate.

Examples of nonionic surfactants include polyoxyethylene alkyl ethers, polyoxyethylene alkylphenyl ethers, polyoxyethylene higher fatty acid esters, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene glycerin fatty acid esters, polyoxyethylene hydrogenated castor oil, coconut oil fatty acid diethanolamide, polyoxyethylene polyoxypropylene alkyl ethers, fatty acid alkanolamides, and alkylamine oxides.

Examples of amphoteric surfactants include betaine surfactants. Specific examples include lauryl-N,N-dimethylacetate betaine, lauryl amidopropyl-N,N-dimethylacetate betaine, and coconut alkyl amidopropyl-N,N-dimethylhydroxypropyl sulfobetaine.

The virus inactivating composition of the present invention is an aqueous type, and water is mainly used as the solvent. Examples of water include purified water such as ion-exchanged water and reverse osmosis water, ordinary tap water, industrial water, deep sea water, etc.

Furthermore, the virus inactivator composition of the present invention may contain other ingredients, such as inorganic antibacterial agents, organic antibacterial agents, virus inactivators, anti-algae agents, anti-rust agents, solvents, chelating agents, fragrances, deodorizing components, pH adjusters, etc., as necessary, within a range that does not impair the effects of the present invention, thereby imparting antibacterial effects, virus inactivation effects, anti-algae effects, anti-rust effects, cleaning effects, fragrances, deodorizing properties, etc.

Examples of inorganic antibacterial agents, organic antibacterial agents, and virus inactivators include isopropylmethylphenol (IPMP), carvacrol, thymol, triclosan, methylparaben, ethylparaben, propylparaben, butylparaben, 4-chloro-3,5-dimethylphenol, orthophenylphenol, o-cresol, m-cresol, p-cresol, tebuconazole, enilconazole, grapefruit seed extract, persimmon seed extract, grape seed extract, monolaurin, monocaprin, monocaprylin, benzoic acid, sorbic acid, glycine, alkyldiethylaminoglycine, polylysine, dehydroacetic acid, chloramine, 3-iodo-2-propyl-N-butylcarbamate (IPBC), phenoxyethanol, silver zeolite, zinc pyrithione, thiamine lauryl sulfate, milt protein, hydroxyalkylchitosan, and chitosan.

An example of an anti-algae agent is sodium dichloroisocyanurate. An example of an anti-rust agent is sodium benzoate.

Examples of solvents include hydrocarbon-based solvents such as normal paraffin, isoparaffin, liquid paraffin, naphthenic hydrocarbons, petrolatum, squalane, and α-olefin oligomers; alcohol-based solvents such as 1-propanol, 2-propanol (IPA), 1-butanol, 2-butanol, tertiary butanol, 1-pentanol, 1-hexanol, benzyl alcohol, and 2-phenylethanol; and glycol-based solvents such as 2-phenoxyethanol (ethylene glycol monophenyl ether), ethylene glycol, propylene glycol, 1-phenoxy-2-propanol (propylene glycol phenyl ether), 1,3-butylene glycol, propylene glycol monobutyl ether, dipropylene glycol monobutyl ether, and tripropylene glycol monobutyl ether.

Examples of fragrances include limonene such as d-limonene, pinene such as α-pinene and β-pinene, cymene such as p-cymene, hydrocarbon fragrances such as indene and caryophyllene, menthol such as linalool, geraniol, citronellol, and l-menthol, ethyl linalool, borneol, anise alcohol, β-phenethyl alcohol, p-menthane-3,8-diol, terpineol such as α-terpineol and γ-terpineol, 1-hexenol, cis-3-hexen-1-ol, and tetrahydrogeraniol. alcohol-based fragrances such as ethyl aldehyde, santalinol, cinnamyl alcohol, cedrol, etc.; ether-based fragrances such as galaxolide, β-naphthyl methyl ether, cineole, ambroxide, p-cresil methyl ether, etc.; phenol-based fragrances such as anethole, eugenol, isoeugenol, vanillin, ethyl vanillin, etc.; octanal, nonanal, undecyl aldehyde, undecanal, decyl aldehyde, n-butyraldehyde, isobutyraldehyde, hexyl aldehyde, citral, citronellal , aldehyde-based fragrances such as benzaldehyde, cinnamic aldehyde, anisaldehyde, cuminaldehyde, adoxal, amyl cinnamic aldehyde, and cyclamen aldehyde, musk ketone, carvone, menthone, camphor, acetophenone, butyrophenone, tonalide, α-ionone, β-ionone, α-methyl ionone, β-methyl ionone, α-isomethyl ionone, β-isomethyl ionone, γ-methyl ionone, γ-isomethyl ionone, damascone, α-damascone, β-damascone Ketone-based fragrances such as γ-butyl lactone, γ-nonalactone, γ-decalactone, γ-undecalactone, coumarin, cineole, ambrosia lid, jasmolactone, and other lactone-based fragrances; geranyl formate, octyl acetate, geranyl acetate, benzyl acetate, cinnamyl acetate, tetrahydrogeranyl acetate, menthyl acetate, linalyl acetate, butyl propionate, benzyl acetate, methyl benzoate, Allyl, allyl hexanoate, allyl heptanoate, allyl cyclohexane propionate, allyl amyl glycolate, amyl valerianate, amyl salicylate, isoamyl acetate, butyl acetate, ethyl butyrate, acetyl eugenol, isoamyl salicylate, allyl caproate, ethyl caproate, ethyl propionate, ethyl acetoacetate, methyl salicylate, citronellyl acetate, citronellyl formate, cinnamyl acetate, stearyl acetate , ester-based fragrances such as stearyl propionate, cedryl acetate, and terpinyl acetate, acetal-based fragrances such as amyl cinnamic aldehyde dimethyl acetal and citral dimethyl acetal, indole, geranyl nitrile, citronellyl nitrile, acetaldehyde phenylethyl propyl acetate, thesalon, auranthiol, linalool oxide, peppermint oil, orange oil, lemon oil, lavender oil, peppermint oil, eucalyptus oil, citronella oil, lime oil, yuzu oil, jasmine oil, Cypress oil, green tea essential oil, neroli oil, geranium oil, petitgrain oil, lemongrass oil, cinnamon oil, lemon eucalyptus oil, thyme oil, perilla oil, pine oil, rose oil, rosemary oil, camphor oil, aromatic oil, clary sage oil, sandalwood oil, spearmint oil, star anise oil, lavandin oil, oakmoss oil, ocotia oil, patchouli oil, tonka bean tincture, turpentine oil, crocodile bean tincture, basil oil, nutmeg oil, clove oil, boreal rose oil, cananga oil, cardamom oil, cassia oil, cedarwood oil, mandarin oil, Examples of oils that may be used include tangerine oil, anise oil, bay oil, coriander oil, elemi oil, fennel oil, galbanum oil, cypress oil, vetiver oil, bergamot oil, ylang-ylang oil, grapefruit oil, abies oil, accion oil, almond oil, angelica root oil, peper oil, mint oil, perch oil, boerboel oil, kayabuchi oil, gananga oil, capsicum oil, caraway oil, celery oil, cognac oil, cumin oil, jell oil, estgolan oil, garlic oil, ginger oil, hops oil, sage oil, and turpentine oil.

Examples of deodorizing ingredients include sugarcane extract, green tea extract, tea distillate, persimmon extract, grapefruit extract, moso bamboo extract, yuzu seed extract, and forsythia extract, but sugarcane extract is preferred from the standpoint of promoting the inactivation of norovirus in addition to its deodorizing effect.

Examples of pH adjusters include other organic acids such as acetic acid, malic acid, salicylic acid, other inorganic acids such as hydrochloric acid, sodium citrate, sodium carbonate, sodium bicarbonate, sodium hydroxide, etc.

The virus inactivator composition of the present invention thus obtained can be applied or sprayed to areas that have been touched by a virus-infected person, areas where vomit from a virus-infected person has been disposed of, clothing, and other virus-contaminated areas, thereby effectively removing the virus. Furthermore, the virus inactivator composition of the present invention is extremely highly practical because it can be applied safely and simply by limiting the blending amount of flammable and irritating ethanol to 10-60% by mass.

The virus inactivating composition of the present invention has a high inactivation effect not only on enveloped viruses such as influenza virus, coronavirus, and herpes virus, but also on non-enveloped viruses such as norovirus, rotavirus, rhinovirus, and adenovirus. Therefore, it can be suitably used to inactivate norovirus, which was difficult to inactivate with conventional virus removal agents.

The virus inactivator composition of the present invention has a very simple composition in which ethanol as a virus inactivating component and a specific organic acid and/or inorganic acid as a virus inactivation efficacy enhancing component are mixed with water to form an acidic aqueous solution. Therefore, it is easy to manufacture, and since it has a low ethanol concentration of 60 mass% or less and is acidic, it has little odor of ethanol and little irritation to skin, which are problems with many virus inactivators, and is extremely safe. In addition, it is also easy to use as there is no problem of discoloration due to silver ions.

The present invention is not limited to the above-described embodiments, and various modifications are possible within the scope of the claims. The technical scope of the present invention also includes embodiments obtained by appropriately combining the technical means disclosed in the different embodiments. The effects of the present invention will be explained in more detail below using examples, but the present invention is not limited to these examples.

[Preparation of test solution]
(A) Ethanol (manufactured by Wako Pure Chemical Industries, Ltd.), (B) fumaric acid, phosphoric acid, and citric acid monohydrate (all manufactured by Wako Pure Chemical Industries, Ltd.) were mixed in the mixing ratios (mass%) shown in Tables 1 and 3, and purified water was added to make up 100 mass%, to obtain test solutions (present inventions 1 to 12).

(A) Ethanol (manufactured by Wako Pure Chemical Industries, Ltd.) (B) Fumaric acid, phosphoric acid, citric acid monohydrate, lactic acid, benzoic acid, trisodium phosphate, trisodium citrate dihydrate (all manufactured by Wako Pure Chemical Industries, Ltd.), and disodium fumarate (manufactured by Tokyo Kaseihin Kogyo Co., Ltd.) were mixed in the proportions (mass %) shown in Tables 2 and 4, and purified water was added to make up 100 mass % to obtain test solutions (Comparative Examples 1 to 16).

[Confirmation test 1 for virus inactivation effect (Feline calicivirus)]
(Preparation of test virus solution)
CRFK cells (JCRB cell bank) were cultured in a monolayer in a tissue culture dish using a cell growth medium made of MEM medium (manufactured by Nacalai Tesque) supplemented with 10% fetal bovine serum. The cell growth medium was removed from the monolayer culture dish, and feline calicivirus (Feline calicivirus F-9 ATCC VR-782) was inoculated. Next, a cell maintenance medium made of MEM medium supplemented with 2% fetal bovine serum was added, and the cells were cultured for 1 to 5 days in a carbon dioxide incubator ( _CO2 concentration 5%) at 37±1°C. Feline calicivirus is a type of non-enveloped virus, and is widely used as an alternative to norovirus, which cannot be cultured in cells.

After the culture, the cell morphology was observed using an inverted phase contrast microscope to confirm that the cells had undergone morphological changes (cytopathic effect). Next, the culture solution was centrifuged at 1000 rpm for 3 minutes, and the resulting supernatant was ultrafiltered to obtain the test virus solution.

0.1 mL of the test virus solution was added to 0.9 mL of the test solutions of Inventions 1 to 7 and Comparative Examples 1 to 11 prepared in Example 1 and mixed to prepare working solutions. After 1 minute, the working solutions were diluted 100-fold with MEM medium to prepare a 10-fold dilution series. Note that the same procedure was carried out using purified water to which the test virus solution was added as a control.

(Measurement of virus infectivity)
The cells used were cultured in a monolayer in a tissue culture microplate (96 wells) using a cell growth medium, and then the cell growth medium was removed and 0.1 mL of cell maintenance medium was added to each well. Next, 0.1 mL of a 10-fold dilution series of the working solution was inoculated into each of 4 wells, and the cells were cultured for 4 to 7 days in a carbon dioxide incubator ( _CO2 concentration 5%) at 37±1°C. After the culture, the presence or absence of morphological changes (cytopathic effect) of the cells was observed using an inverted phase contrast microscope, and the 50% cell culture infectious dose ( _TCID50 ) was calculated by the Reed-Muench method and converted into an infectious titer per 1 mL of the working solution. The infectious titer log reduction was calculated by comparing with the infectious titer of purified water used as a control.

The evaluation criteria for the virus removal effect were as follows: × when the log reduction in infectivity value was 1 or less, △ when it was more than 1 and 1.5 or less, ○ when it was more than 1.5 and 3 or less, and ◎ when it was more than 3. The evaluation results of the virus infectivity value are shown in Tables 1 and 2 together with the composition of the test solution, pH, log ₁₀ [(A)/(B)], and the log reduction in infectivity value.

As shown in Table 1, in the compositions 1 to 7 of the present invention, which contain (A) 10% to 40% by mass of ethanol and (B) 0.05% to 0.5% by mass of either fumaric acid, phosphoric acid, or citric acid monohydrate, the log reduction in infectivity against feline calicivirus was greater than 1.5, confirming that the compositions have a virus inactivation effect.

In particular, in Inventions 1 and 2, in which the amount of ethanol was 40% by mass and the amount of fumaric acid was 0.2% by mass and 0.1% by mass, respectively, and Invention 5, in which the amount of ethanol was 40% by mass and the amount of phosphoric acid was 0.05% by mass, the infectivity log reduction value was greater than 3, and it was confirmed that a more significant virus inactivation effect was obtained. Furthermore, from the results of Inventions 1 to 7 and Comparative Examples 1 to 5, it was confirmed that when ethanol was blended alone, or when fumaric acid, phosphoric acid, or citric acid monohydrate was blended alone, the infectivity log reduction value was less than 1 and no virus inactivation effect was observed, but that the virus inactivation effect was synergistically enhanced by blending ethanol with fumaric acid, phosphoric acid, or citric acid monohydrate.

Furthermore, from the results of Invention 6, in which the ethanol content was 10% by mass and the phosphoric acid content was 0.5% by mass, and Comparative Example 8, in which the ethanol content was 5% by mass and the phosphoric acid content was 0.5% by mass, it was confirmed that by making the ethanol content 10% by mass or more, the viral inactivation effect was synergistically enhanced by the inclusion of phosphoric acid.

Furthermore, from the results of Invention 5, in which the ethanol content is 40% by mass and the phosphoric acid content is 0.05% by mass, and Comparative Example 7, in which the ethanol content is 40% by mass and the phosphoric acid content is 0.02% by mass, it was confirmed that by making the phosphoric acid content 0.05% by mass or more, the virus inactivation efficacy of ethanol is enhanced.

In addition, in Comparative Examples 9 to 11, in which lactic acid, benzoic acid, and disodium fumarate were used at 0.2 mass% instead of fumaric acid, phosphoric acid, and citric acid monohydrate, the log reduction in infectivity was 1.5, 0.7, and 1.33, respectively, and sufficient virus inactivation effect was not observed.

[Confirmation test 2 of virus inactivation effect (Feline calicivirus)]
(Preparation of test virus solution)
CRFK cells (JRBC cell bank) were cultured in a monolayer in a tissue culture dish using a cell growth medium made of MEM medium (manufactured by Nacalai Tesque) with 10% fetal bovine serum. The cell growth medium was removed from the monolayer culture dish, and feline calicivirus (Feline calicivirus F-9 ATCC VR-782) was inoculated. Next, a cell maintenance medium made of MEM medium with 2% fetal bovine serum was added, and the cells were cultured in a carbon dioxide gas incubator ( _CO2 concentration 5%) at 37±1°C for 1 to 5 days.

After culturing, the cell morphology was observed using an inverted phase contrast microscope, and it was confirmed that the cells had undergone morphological changes (cytopathic effect). Next, the culture solution was centrifuged at 1000 rpm for 3 minutes, and the resulting supernatant was used as the virus suspension. The virus suspension and a filter-sterilized 20% meat extract aqueous solution were mixed in a ratio of 3:1 to prepare the test virus solution. The test virus solution contained 5% meat extract as a load.

0.1 mL of the test virus solution was added to 0.9 mL of the test solutions of Invention Nos. 8 to 12 and Comparative Examples 12 to 16 prepared in Example 1 and mixed to prepare working solutions. After 1 minute, the working solutions were diluted 100-fold with MEM medium to prepare a 10-fold dilution series. Note that the same procedure was carried out using purified water to which the test virus solution was added as a control.

The method for measuring the virus infectivity and the evaluation criteria were the same as in Example 2. The evaluation results of the virus infectivity are shown in Tables 3 and 4 together with the formulation of the test solution, pH, log ₁₀ [(A)/(B)], and the log reduction in the infectivity.

As shown in Table 3, in the compositions of inventions 8 to 12, which contain (A) 40% to 60% by mass of ethanol and (B) 0.05% to 0.2% by mass of either fumaric acid, phosphoric acid, or citric acid monohydrate, the log reduction in infectivity against feline calicivirus was 2.5 or more, confirming that an immediate virus inactivation effect can be obtained even in the presence of a load of meat extract.

In contrast, as shown in Table 4, in Comparative Example 12, in which ethanol was used alone at 60% by mass, and in Comparative Example 13, in which the amount of ethanol was 60% by mass and the amount of phosphoric acid was 0.02% by mass, the infectivity log reduction value was less than 1, and no virus inactivation effect was observed. In Comparative Examples 14 and 15, in which disodium fumarate and trisodium phosphate were used instead of fumaric acid and phosphoric acid, the infectivity log reduction values were 1.43 and 1.24, respectively, and no sufficient virus inactivation effect was observed. Furthermore, in Comparative Example 15, the pH of the test solution was 12.51, which was strongly alkaline, raising concerns about alkaline corrosiveness and rough hands.

In addition, in Comparative Example 16, in which trisodium citrate was used instead of citric acid monohydrate, the log reduction in infectivity was 1.93, indicating a virus inactivation effect; however, the pH of the test liquid was alkaline at 9.15, raising concerns about the corrosiveness and rough hands caused by the alkali.

From the above results, it was confirmed that by mixing ethanol and one or more selected from fumaric acid, phosphoric acid, and citric acid monohydrate with water to prepare an acidic aqueous solution, a virus inactivator composition that has sufficient virus inactivation effect against feline calicivirus and is also highly safe can be obtained.

It was also confirmed that when combined with ethanol, fumaric acid, phosphoric acid, and citric acid monohydrate act as ingredients that enhance the virus inactivation efficacy of ethanol.

Although the inactivation effect of the virus inactivation composition of the present invention on enveloped viruses is not shown here, since enveloped viruses are more sensitive to drugs than non-enveloped viruses, it goes without saying that the virus inactivation composition of the present invention also has an immediate inactivation effect on enveloped viruses.

The present invention relates to a virus inactivator composition, a method for enhancing virus inactivation efficacy, and a virus inactivation method that have a relatively low concentration of ethanol, a virus inactivating component, and are acidic to weakly acidic and therefore highly safe, and is particularly suitable for use as a virus inactivator composition that is directly applied by spraying or the like to places contaminated with viruses.

Claims (2)

(A) 10% by mass to 60% by mass of ethanol as a virus inactivation component, (B) fumaric acid as a virus inactivation effect enhancing component, and (C) water are mixed to prepare an acidic aqueous solution;
A virus inactivator composition comprising the virus inactivation efficacy enhancing component in an amount of 0.05 mass% or more , the virus inactivation efficacy enhancing component being targeted at non-enveloped viruses . (A) a virus inactivating agent composition containing 10% by mass to 60% by mass of ethanol as a virus inactivating component and (C) water, and (B) fumaric acid as a virus inactivation efficacy enhancing component is added to the virus inactivating agent composition;
A method for enhancing virus inactivation efficacy, characterized in that the amount of the virus inactivation efficacy enhancing component added is 0.05 mass% or more relative to the virus inactivator composition, and the method is directed to non-enveloped viruses .