Antifungal composition

Abstract

A composition is described comprising an anionic surfactant, especially sodium dodecyl sulphate, an organic acid, especially lactic acid, and one or more compounds of Formula I, especially trans-anethole and / or estragole. The composition may find use in an antifungal or fungicidal application.

Description

[0001] ANTIFUNGAL COMPOSITION

[0002] BACKGROUND

[0003] Moulds (or “molds” in the USA) are multicellular filamentous fungi which can grow in many environments, including inside households under certain conditions. Mould may grow on surfaces such as a floor or a wall, and may also grow on surfaces of objects or devices, especially where those objects or devices come into contact with moisture or are difficult to dry. For example, cleaning tools or appliances may eventually become colonised by mould if not fully dried. When mould spores are present in large quantities, they can present a health hazard to humans, potentially causing allergic reactions and respiratory problems. Furthermore, mould within cleaning devices may lead to unpleasant smells and may cause the spread of fungal spores during subsequent use of the device.

[0004] It is traditional to use household bleach to attempt to remove or kill fungus, such as mould. This involves treatment of the surface on which the mould has become established with the composition containing bleach. However bleach may be unsuitable for use on certain household surfaces, and may cause damage to those surfaces.

[0005] Furthermore, many existing disinfectant compositions are only suitable for use on precleaned surfaces, or are less effective on “dirty” surfaces, because the presence of organic matter on a surface is known to reduce the efficacy of the compositions in removing or suppressing the growth of unwanted organisms such as fungi.

[0006] SUMMARY

[0007] The invention provides a composition which may be effective for killing fungal spores, in particular spores of fungi which are known to form mould. Without wishing to be bound by theory, such killing of spores may mean that it is less likely that the mould will grow. Furthermore, the composition may be more suitable for use in a wider variety of environments (for example, on a wider range of surfaces) than alternative compositions such as bleach-containing compositions. A first aspect of the invention is a composition comprising an anionic surfactant, an organic acid, and one or more compounds of Formula I:

[0008] Formula I wherein R1is an unsaturated C3-C6 hydrocarbyl group, and R2is a C1-C3 alkyl group.

[0009] Such a composition may be used to treat environments (e.g. surfaces) which have been colonised by fungus, such as mould, and may be effective at killing or rendering inviable the fungal spores within the mould, to reduce the likelihood that the mould will recolonise the environment after removal.

[0010] Furthermore, the composition may be less harsh than bleach, and may therefore be used in a wider variety of environments without the risk of damage, for example damage to a surface.

[0011] A second aspect of the invention is a method of killing or preventing the growth of fungus, comprising contacting the fungus with the composition according to the first aspect.

[0012] Such a method may be effective at preventing the growth or regrowth of fungus, such as mould.

[0013] A third aspect of the invention is the use of a compound according to Formula I as an antifungal or fungicidal agent in a composition:

[0014] Formula I wherein R1is an unsaturated C3-C6 hydrocarbyl group, and R2is a C1-C3 alkyl group.

[0015] Without wishing to be bound by theory, it is believed that the compound according to Formula I may impart the composition with antifungal or fungicidal properties, or enhanced antifungal or fungicidal properties, relative to a composition lacking such a compound.

[0016] A fourth aspect of the invention is a method of manufacturing the composition according to the first aspect.

[0017] FURTHER OPTIONS AND PREFERENCES

[0018] The composition of the first aspect contains one or more compounds according to Formula I:

[0019] Formula I wherein R1is an unsaturated C3-C6 hydrocarbyl group, and R2is a C1-C3 alkyl group.

[0020] The composition may comprise a mixture of different compounds, each selected from compounds according to Formula I. For example, the composition may comprise two different compounds, each selected from compounds according to Formula I.

[0021] By “unsaturated C3-C6 hydrocarbyl group” is meant a hydrocarbyl group containing a total number of carbon atoms in the range 3-6, and containing at least one C=C double bond. The unsaturated C3-C6 hydrocarbyl group may be bound to the phenylene group of the compound according to Formula I by either an sp2or sp3hybridised carbon atom.

[0022] R1may be an unsaturated C3-C5 hydrocarbyl group, for example an unsaturated C3-C4 hydrocarbyl group, or an unsaturated C3 hydrocarbyl group. R1may be an unsaturated straight-chain C3-C6 hydrocarbyl group. In other words, all of the carbon atoms within R1may lie within a single contiguous chain, with no branches.

[0023] R1may be an unsubstituted, unsaturated C3-C6 hydrocarbyl group. R1may be an unsubstituted, unsaturated straight-chain C3-C6 hydrocarbyl group.

[0024] By “C1-C3 alkyl group” is meant a saturated hydrocarbyl group containing a total number of carbon atoms in the range 1-3.

[0025] R2may be an unsubstituted C1-C3 alkyl group.

[0026] R2may be methyl or ethyl. R2may be methyl.

[0027] R2may be unsubstituted methyl or ethyl. R2may be unsubstituted methyl.

[0028] The composition of the first aspect may contain one or more compounds according to Formula I, wherein R1is an unsaturated C3-C4 hydrocarbyl group, and R2is a C1-C2 alkyl group. The composition of the first aspect may contain one or more compounds according to Formula I, wherein R1is an unsaturated C3 hydrocarbyl group, and R2is methyl. The composition of the first aspect may contain one or more compounds according to Formula I, wherein R1is an unsaturated unsubstituted C3 hydrocarbyl group, and R2is methyl.

[0029] The one or more compounds of Formula I may comprise or consist of one or both

[0030] These compounds are known as / ra / / .s-anethole ( ) and estragole ). They are naturally-occurring compounds which are found in plants. They may be found separately or in combination in certain essential oils. For example, / ra / z.s-an ethole is found in fennel oil, and may also be found in aniseed oil. Estragole may be found in basil oil and tarragon oil. Some essential oils may contain both / ra / z.s-anethole and estragole, although one of the two compounds is often dominant.

[0031] The one or more compounds of Formula I may be present in the composition in an amount of at least 0.01 wt%, for example at least 0.015 wt%, at least 0.02 wt%, at least 0.05 wt%, at least 0.08 wt% or at least 0.10 wt%, based on the total weight of the composition.

[0032] The one or more compounds of Formula I may be present in the composition in an amount of up to 5.0 wt%, for example up to 4.0 wt%, up to 3.0 wt%, up to 2.0 wt% or up to 1.0 wt%, based on the total weight of the composition.

[0033] The one or more compounds of Formula I may be present in the composition in an amount of from 0.015 to 1.0 wt%, based on the total weight of the composition, for example from 0.018 to 1.0 wt%, from 0.015 to 0.50 wt%, from 0.018 to 0.50 wt% or from 0.10 to 0.50 wt%.

[0034] Such quantities of the one or more compounds of Formula I may contribute to enhanced fungicidal properties of the composition. They may also allow the quantities of further components of the composition to be reduced, without jeopardising the fungicidal properties of the composition. For example, the presence of the one or more compounds of Formula I may allow the amounts of anionic surfactant and / or organic acid within the composition to be reduced while maintaining good fungicidal properties of the composition.

[0035] The composition may comprise an essential oil component which contains the one or more compounds of Formula I. Certain essential oils are known to contain compounds which have a structure according to Formula I, so including one or more of such essential oils within an essential oil component of the composition is a way to provide the required compound according to Formula I. For example, fennel oil, aniseed oil, basil oil and tarragon oil are all known to contain one or more of / ra / z.s-anethole and estragole. The essential oil component may comprise or consist of one or more individual essential oils. The essential oil component may comprise or consist of a single type of essential oil.

[0036] The essential oil component may comprise or consist of one or more of fennel oil, aniseed oil, basil oil, and tarragon oil.

[0037] The essential oil component may comprise or consist of fennel oil.

[0038] The essential oil component may be present in the composition in an amount to achieve the above-defined quantities of the compound according to Formula I. The amount of essential oil component used may depend on the concentration of the one or more compounds according to Formula I within the chosen essential oil component.

[0039] The essential oil component may be present in the composition in an amount of from 0.05 to 10 wt%, for example from 0.05 to 9 wt%, from 0.05 to 8 wt%, from 0.05 to 7 wt%, from 0.05 to 6 wt%, from 0.05 to 5 wt%, from 0.05 to 2 wt%, from 0.05 to 1 wt% or from 0.1 to 1 wt%.

[0040] The composition of the first aspect contains an anionic surfactant.

[0041] The anionic surfactant may be a sulfate, sulfonate, phosphate or carboxylate surfactant. The anionic surfactant may be a sulfate surfactant. The anionic surfactant may be an alkyl sulfate.

[0042] The anionic surfactant may comprise or consist of a dodecyl sulfate surfactant, for example ammonium dodecyl sulfate (ALS) or sodium dodecyl sulfate (SDS).

[0043] The anionic surfactant may comprise or consist of sodium dodecyl sulphate (SDS). The anionic surfactant may consist of sodium dodecyl sulphate (SDS). The anionic surfactant may be present in the composition in an amount of from 0.1 to 5 wt%, based on the total weight of the composition, for example from 0.2 to 5 wt%, from 0.3 to 5 wt%, from 0.4 to 5 wt%, from 0.5 to 5 wt%, from 0.5 to 4 wt%, from 0.5 to 3 wt%, from 0.5 to 2 wt% or from 0.5 to 1 wt%.

[0044] The composition of the first aspect contains an organic acid.

[0045] The organic acid may comprise or consist of one or more carboxylic acids. The organic acid may comprise or consist of one or more Ci-6 carboxylic acids (i.e. carboxylic acids containing a total number of carbon atoms in the range 1-6).

[0046] The organic acid may comprise or consist of a Ci-6 carboxylic acid comprising at least one additional -OH group aside from any -OH groups within the carboxylate group.

[0047] The organic acid may comprise or consist of one or more of methanoic acid, ethanoic acid, propanoic acid and butanoic acid, each optionally substituted by one or more -OH groups. The organic acid may comprise or consist of one or more of methanoic acid, ethanoic acid and propanoic acid, each optionally substituted by one or more -OH groups. The organic acid may comprise or consist of propanoic acid, optionally substituted by one or more -OH groups.

[0048] The organic acid may comprise or consist of one or more C3 carboxylic acids. The organic acid may comprise or consist of a single type of C3 carboxylic acid. The organic acid may comprise or consist of a C3 carboxylic acid comprising at least one additional -OH group aside from any -OH groups within the carboxylate group.

[0049] The organic acid may comprise or consist of lactic acid (2-hydroxypropanoic acid).

[0050] The organic acid may be present in the composition in an amount of from 1 to 10 vol%, for example 3 to 10 vol%, based on the total volume of the composition. The organic acid may be present in the composition in an amount of from 1.0 to 12.5 wt%, based on the total weight of the composition, for example from 1.0 to 12.0 wt%, from 1.0 to 10.0 wt%, from 1.0 to 8.0 wt%, from 1.0 to 7.0 wt%, from 1.0 to 6.5 wt%, or from 1.0 to 6.5 wt%.

[0051] The organic acid may be present in the composition in an amount of at least 2.5 wt%, for example at least 3.0 wt%, at least 3.5 wt%, at least 4.0 wt% or at least 4.5 wt%, based on the total weight of the composition.

[0052] The organic acid may be present in the composition in an amount of from 2.5 to 12.5 wt%, based on the total weight of the composition, for example from 3.5 to 12.0 wt%, from 3.5 to 10.0 wt%, from 3.5 to 8.0 wt%, from 3.5 to 7.0 wt%, from 3.5 to 6.5 wt%, or from 4.5 to 6.5 wt%.

[0053] The composition of the first aspect comprises the anionic surfactant, the organic acid, and the one or more compounds of Formula I.

[0054] The composition may comprise the anionic surfactant, the organic acid, the one or more compounds of Formula I, and a solvent.

[0055] The composition may comprise the anionic surfactant, the organic acid, the one or more compounds of Formula I, wherein the balance of the composition is a solvent.

[0056] The composition may consist of the anionic surfactant, the organic acid, the one or more compounds of Formula I, and a solvent.

[0057] The solvent may comprise or consist of one or more of water, ethanol, methanol, propanol, ethyl acetate, and acetone.

[0058] The composition may comprise or consist of 0.1 to 5 wt% anionic surfactant, 3.5 to 12.5 wt% organic acid, 0.015 to 1.0 wt% of the one or more compounds of Formula I, and a solvent, based on the total weight of the composition. The composition may comprise or consist of 0.1 to 5 wt% anionic surfactant, 3.5 to 12.5 wt% organic acid, 0.015 to 1.0 wt% of the one or more compounds of Formula I, wherein the balance of the composition is a solvent, based on the total weight of the composition.

[0059] The composition may comprise or consist of 0.1 to 5 wt% SDS, 3.5 to 12.5 wt% lactic acid, 0.015 to 1.0 wt% of one or more of / ra / z.s-anethole and estragole, and a solvent, based on the total weight of the composition. The composition may comprise or consist of 0.1 to 5 wt% SDS, 3.5 to 12.5 wt% lactic acid, 0.015 to 1.0 wt% of of one or more of trans- anethole and estragole, wherein the balance of the composition is a solvent, based on the total weight of the composition.

[0060] The composition may comprise or consist of 0.5 to 1 wt% anionic surfactant, 3.6 to 6.5 wt% organic acid, 0.10 to 0.50 wt% of the one or more compounds of Formula I, and a solvent, based on the total weight of the composition. The composition may comprise or consist of 0.5 to 1 wt% anionic surfactant, 4.5 to 6.5 wt% organic acid, 0.10 to 0.50 wt% of the one or more compounds of Formula I, wherein the balance of the composition is a solvent, based on the total weight of the composition.

[0061] The composition may comprise or consist of 0.5 to 1 wt% SDS, 3.6 to 6.5 wt% lactic acid, 0.10 to 0.50 wt% of one or more of / ra / z.s-anethole and estragole, and a solvent, based on the total weight of the composition. The composition may comprise or consist of 0.5 to 1 wt% SDS, 4.5 to 6.5 wt% lactic acid, 0.10 to 0.50 wt% of one or more of / ra / z.s-anethole and estragole, wherein the balance of the composition is a solvent, based on the total weight of the composition.

[0062] The composition may comprise at least 0.5 wt% SDS, at least 2.5 wt% lactic acid and at least 0.05 wt% of an essential oil component comprising one or more of fennel oil, aniseed oil, basil oil, and tarragon oil.

[0063] The composition may comprise from 1.0 to 2.0 wt% SDS, at least 4.5 wt% lactic acid and at least 1.0 wt% of an essential oil component comprising one or more of fennel oil, aniseed oil, basil oil, and tarragon oil. The composition may be in liquid form. The composition may be an aqueous solution of the anionic surfactant, organic acid, and one or more compounds of Formula I.

[0064] The composition may contain one or more further additives. The amount of any further additives may be up to 5 wt% of the composition, for example up to 4 wt%, up to 3 wt%, up to 2 wt% or up to 1 wt%.

[0065] For example, the composition may comprise an alcohol, such as ethanol. The alcohol may be present in an amount of up to 5 wt% of the composition, for example up to 4 wt%, up to 3 wt%, up to 2 wt% or up to 1 wt%, or from 0.5 to 2 wt%. The alcohol may assist in the dissolution or dispersion of the essential oil component in the composition.

[0066] Other optional additives may include one or more of fragrance, colorant, thickener, pH adjuster and antioxidant. Other antimicrobial, such as antifungal or antibacterial, agents may also be included in the composition.

[0067] The second aspect of the invention is a method of killing or preventing the growth of fungus, comprising contacting the fungus with the composition according to the first aspect.

[0068] The fungus may comprise or consist of mould.

[0069] The ability of a composition to kill fungal spores or prevent the growth of mould as described herein may be characterised by the “log reduction” (R) in fungal spore growth or concentration. The log reduction, R, represents the extent to which the composition is able to kill the spores and prevent their subsequent proliferation. The R for a composition is defined as the base- 10 logarithm of the ratio of the number of fungal spores before and after exposure to the composition. So, for example, a “1-log reduction” represents a 10- fold (i.e. 90%) decrease in the number of spores, a “2-log reduction” represents a 100-fold (i.e. 99%) reduction in the number of spores, and so on. Many compositions of the invention may achieve at least a 4-log (i.e. 99.99%) reduction in the number of spores. The method of the second aspect may provide at least a log-2 reduction in the number or concentration of fungal spores, for example at least a log-2.5, at least a log-2.6, at least a log-2.7, at least a log-2.8, at least a log-2.9, at least a log-3, at least a log-3.5, or at a least log-4 reduction.

[0070] R may be determined by exposing a sample containing a test species such as Aspergillus brasiliensis (ATCC 16404) spores to the composition for 15 minutes, neutralising the composition, then incubating the sample for 48 hours at 30 °C in a malt extract agar plate and then comparing the number of spores with a benchmark agar plate containing spores incubated under the same conditions which have not been exposed to the composition.

[0071] Aspergillus brasiliensis (ATCC 16404) spores are considered to be a challenging organism, such that efficacy against Aspergillus brasiliensis (ATCC 16404) spores can be considered indicative of a highly fungicidal composition.

[0072] For example, the method in EN 1650:2019, “Chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of fungicidal activity of chemical disinfectants and antiseptics used in food, industrial, domestic, and institutional areas”, may be applied to determine R for a composition of the invention.

[0073] The method of the second aspect may result in at least a 2-log reduction in the number of Aspergillus brasiliensis (ATCC 16404) spores per mL, for example at least a log-2.5, at least a log-2.6, at least a log-2.7, at least a log-2.8, at least a log-2.9, at least a log-3, at least a log-3.5, or at least a log-4 reduction.

[0074] The method of the second aspect may comprise applying the composition according to the first aspect to an environment which contains mould. The method of the second aspect may comprise applying the composition according to the first aspect to an environment which is colonised by mould. The method of the second aspect may comprise applying the composition according to the first aspect to a surface. The method of the second aspect may comprise applying the composition to a surface without any pre-treatment of the surface, for example without any preliminary cleaning of the surface. Many known fungicidal compositions require pre-cleaning of surfaces, because the efficacy of the composition is reduced in the presence of organic contaminants. The compositions of the invention may provide improved fungicidal activity on “dirty” surfaces containing e.g. organic contaminants, without the need for an earlier cleaning step.

[0075] The method may comprise contacting the mould with the composition, to do one or more of (a) remove the mould, (b) prevent further proliferation of the mould, and (c) prevent regrowth of the mould after removal.

[0076] The method may comprise applying the composition to a surface containing a mould colony, to do one or more of (a) remove the mould from the surface, (b) prevent further proliferation of the mould, and (c) prevent regrowth of the mould after removal.

[0077] The method may comprise applying the composition to an environment (e.g. a surface) which does not contain any mould, to reduce the likelihood of any future growth of mould in the environment.

[0078] The method may comprise applying the composition to the surface, allowing the composition to remain in contact with the surface for a period of time, and rinsing the surface to remove the composition. The period of time may be e.g. at least 30 seconds, at least 1 minute or at least 5 minutes.

[0079] The surface may be a floor, a wall or a work-surface. The surface may be a surface of a household object, for example a toy. The surface may be a surface of a device. The surface may be an interior surface of a cleaning device. The surface may be an interior surface of a household cleaning device, where the interior surface comes into contact with moisture during use of the household cleaning device. The method may therefore comprise contacting the interior surface of the device with the composition to sanitise the device, for example to remove any existing mould growth and / or prevent further mould growth within the device. Administration of the composition onto the surface may be done by any suitable application method. For example, the composition may be applied to an applicator such as a mop, sponge or cloth, before wiping the surface with the applicator. The composition may be poured or sprayed directly onto the surface. Where the surface is an interior surface of a cleaning device, the composition may be poured or injected into the device, may be added to the cleaning fluid which is ordinarily used within the device, or may be used in place of the cleaning fluid which is ordinarily used within the device, for an occasional mould treatment procedure.

[0080] The third aspect of the invention is the use of a compound according to Formula I, as defined under the first aspect, as an antifungal or fungicidal agent in a composition.

[0081] The options set out under the first aspect above for the variables within Formula I apply equally to the third aspect, but for brevity are not repeated here.

[0082] The options set out under the first aspect above for the amounts of the compound according to Formula I within the composition apply equally to the third aspect, but for brevity are not repeated here.

[0083] The composition may further comprise an anionic surfactant and an organic acid. The options set out under the first aspect above for the identities and quantities of the anionic surfactant and organic acid apply equally to the third aspect, but for brevity are not repeated here.

[0084] The use of the third aspect may comprise the use to achieve at least a log-2 reduction in the number or concentration of fungal spores, for example at least a log-2.5, at least a log-2.6, at least a log-2.7, at least a log-2.8, at least a log-2.9, at least a log-3, at least a log-3.5, or at least a log-4 reduction.

[0085] The use of the third aspect may comprise the use to achieve at least a 2-log reduction in the number of Aspergillus brasiliensis (ATCC 16404) spores per mL, for example at least a log-2.5, at least a log-2.6, at least a log-2.7, at least a log-2.8, at least a log-2.9, at least a log-3, at a least log-3.5, or at least a log-4 reduction.

[0086] Such log reductions may be determined by the methods set out above under the second aspect.

[0087] The use of the third aspect may comprise the use to provide one or more of:

[0088] (a) removing mould;

[0089] (b) preventing further proliferation of mould; and

[0090] (c) reducing the likelihood of future growth of mould.

[0091] The fourth aspect is a method of manufacturing the composition according to the first aspect.

[0092] The method may comprise combining the anionic surfactant, the organic acid and the one or more compounds of Formula I; and mixing to form the composition.

[0093] The method may comprise: adding the anionic surfactant to a solvent, such as water, to form a first mixture; stirring or agitating the first mixture; adding the one or more compounds according to Formula I to the first mixture to form a second mixture; stirring or agitating the second mixture; adding the organic acid to the second mixture to form a third mixture; and stirring or agitating the third mixture.

[0094] The stirring or agitating of the first mixture may comprise stirring, for example with a stirrer bar.

[0095] The stirring or agitating of the third mixture may comprise sonication. The one or more compounds according to Formula I may be provided within an essential oil component, which is added to the first mixture. The essential oil component may comprise or consist of an essential oil dissolved in a solvent, such as ethanol.

[0096] The stirring or agitating to form the first mixture may be performed for at least 1 minute, for example at least 2 minutes, at least 5 minutes or at least 10 minutes.

[0097] The stirring or agitating to form the second mixture may be performed for at least 10 seconds, for example at least 30 seconds, at least 40 seconds or at least 60 seconds.

[0098] The stirring or agitating to form the second mixture may comprise a first step of stirring for at least 10 seconds, for example at least 30 seconds, at least 40 seconds or at least 60 seconds; and a second step of sonication for at least 1 minute, for example at least 2 minutes, at least 5 minutes, at least 10 minutes or at least 20 minutes.

[0099] EXAMPLES

[0100] Example 1

[0101] Compositions S1-S18 and C1-C12 were prepared having the ingredients set out in Tables 1-4. In each case the balance of the composition was water.

[0102] Compositions S1-S18 demonstrate the invention, whereas compositions C1-C12 are comparative compositions.

[0103] Table 1 Table 2

[0104] Table 3 Table 4

[0105] Composition C9 would not form a homogeneous mixture

[0106] The compositions were prepared according to the following method:

[0107] SDS powder added to a sterile beaker containing a sterile stirrer bar. The required volume of water was added and mixed on magnetic stir plate at 350 rpm for 10 minutes.

[0108] Meanwhile, the appropriate essential oil was either provided neat, or dissolved in the ethanol for compositions containing ethanol.

[0109] The essential oil (or essential oil in ethanol) was then added to the SDS solution in the beaker and was stirred for 60 seconds.

[0110] Lactic acid was then added to beaker and the solution was stirred for a further 60 seconds.

[0111] The beaker was then placed in a sonicating water bath and the mixture was sonicated for 20 minutes.

[0112] Three commercially available fennel oils, FEO1, FEO2 and FEO3, were obtained and used as indicated in the Tables above. FEO1, FEO2 and FEO3 were subjected to GC-MS analysis to confirm the composition of the oils. For all three oils, / ra / z.s-an ethole and estragole were identified as major components. The amounts of / ra / z.s-an ethole and estragole in each of the oils was as follows (based on GC-MS peak area analysis): Table 5

[0113] Three commercially available aniseed oils, AE01, AEO2 and AEO3, were obtained and used in some of the compositions as indicated in the Tables above. Aniseed oil is known to contain high levels of / ra / z.s-anethole and low levels of estragole.

[0114] Three commercially available basil oils, BEO1, BEO2 and BEO3, were obtained and used in some of the compositions as indicated in the Tables above. Basil oil is known to contain estragole.

[0115] Tarragon oil, which was also used, is known to contain high levels of estragole with little / ra / 7.s-an ethole.

[0116] Example 2 - fungicidal testing

[0117] The ability of the compositions to kill fungal spores was tested.

[0118] The test used was based on the method described by EN 1650:2019, “Chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of fungicidal activity of chemical disinfectants and antiseptics used in food, industrial, domestic, and institutional areas" .

[0119] 1 mL of a Test Suspension was first mixed with 1 mL of Interfering Substance A or Interfering Substance B. The Test Suspension contained spores of Aspergillus brasiliensis ATCC 16404, ensuring a density of spores within the range 1.5 x 107- 5 x 107. Interfering Substance A was a solution of 0.3 g / L bovine serum albumin fraction V, to represent “clean conditions”, and Interfering Substance B was a solution of 3.0 g / L bovine serum albumin fraction V, to represent “dirty conditions”. The Test Suspension and Interfering Substance were mixed for 2 minutes.

[0120] 8 mL of the composition from Example 1 to be tested for fungicidal properties was then added to the mixture. The composition from Example 1 was prepared for testing with 1.25x the concentration of ingredients relative to those provided in Tables 1-4 above, as required by EN 1650, to allow for the dilution when combined in the mixture. The mixture was then allowed 15 minutes contact time, before a 1 mL sample was extracted and combined with 1 mL water and 8 mL neutralising solution, to neutralise the active ingredients.

[0121] The neutralising solution was a mixture that contains substances known to inactivate various classes of biocides. The dilution of the actives into the neutralising solution also reduces the concentration of actives in the mixture. The neutralising solution was validated to confirm that it is effective according to the method described in EN1650. The composition of the neutraliser was the first composition described in Annex B of EN 13697.

[0122] After 5 minutes of neutralisation time, two separate 1 mL samples of the neutralised solution were extracted. The two samples were added to two separate, identical malt extract agar plates.

[0123] A third plate was prepared by adding 1 mL of the untreated Test Suspension containing fungal spores to a third, identical malt extract agar plate.

[0124] The plates were incubated for 48 hours at 30 °C and then the fungal growth counted and the logio reduction in the number of spores per mL for the inoculated sample, relative to the untreated sample, was calculated.

[0125] The results are set out in Table 6, based on an average count from the duplicate agar plates. Table 6

[0126] “7?” in Table 6 is the logio reduction in the number of spores, as defined herein. An R of 4 or greater indicates a 10,000-fold reduction in the number of spores, i.e. a 99.99% reduction.

[0127] The symbol “<” in Table 6 indicates that the fungal growth exceeded the upper limit for counting of fungi on agar plates.

[0128] The symbol “>” in Table 6 indicates that the fungal growth was below the lower limit for the counting of fungi on agar plates. Compositions S1-S18, all containing SDS, lactic acid and an essential oil component known to contain one or both of trans-an ethole and estragole, all demonstrated at least acceptable fungicidal properties, with many approaching or exceeding a 4-log reduction in the number of spores (10,000-fold reduction).

[0129] S4 and S5 demonstrated slightly lower reductions than the other inventive examples, which is thought to be due to the lower levels of lactic acid and / or SDS in those compositions. Nevertheless, fungicidal activity approaching a 3 -log reduction was achieved despite the low levels of these components.

[0130] Some of the comparative compositions including C4, C6 and C7 achieved higher reductions than S4 and S5, but it is inappropriate to compare any of C4, C6 or C7 directly with S4 or S5 due to the differing levels of lactic acid and / or SDS between these compositions. Each of C4, C6 and C7 contain high levels of SDS and lactic acid. Without wishing to be bound by theory, it is believed that the high levels of SDS and lactic acid allow some fungicidal activity to be achieved. However the activity of C4, which contained thyme oil (which does not contain any / ra / z.s-anethole or estragole) is significantly lower than SI, which contains the same amounts of SDS and lactic acid as C4 but with fennel oil (which contains / ra / z.s-anethole and estragole) instead of thyme oil. C6 and C7 contained only SDS and lactic acid, without any essential oil component. Comparing SI with C6 or C7 shows that the addition of fennel oil while maintaining the levels of SDS and lactic acid improves the fungicidal properties. Furthermore, comparing S8 or S9 with C6 or C7 shows that the presence of fennel oil allows the amount of SDS to be reduced in the composition, while providing an increase in fungicidal properties.

[0131] C1-C5 contain essential oil components which do not contain any / ra / z.s-anethole or estragole. Comparing these with SI, it is evident that the presence of / ra / z.s-an ethole or estragole provides a significant increase in fungicidal properties.

[0132] Three different commercially available fennel oils, FEO1, FEO2 and FEO3, were tested (in compositions S9-S11 respectively). The results show that each of these compositions achieved a significant log reduction in fungal spore concentration. Compositions S12-S18 were analogous to compositions S9-S11, with the same quantities of SDS and lactic acid present, but with fennel oil replaced with aniseed oil, basil oil or tarragon oil respectively. Each of aniseed oil, basil oil and tarragon oil are known to contain one or both of / ra / z.s-anethole and estragole. Compositions S12-S18 all achieved a greater than 4-log reduction in the fungal spore concentration.

[0133] Composition C9 (lactic acid and fennel oil) could not form a stable composition, and compositions C10-C12 performed poorly in the fungicidal testing with very low R values. This shows that SDS or lactic acid alone cannot provide the fungicidal properties observed for the compositions of the invention, and that alternative surfactants, including non-ionic surfactants such as Tween, also cannot provide the fungicidal properties observed for the compositions of the invention.

Claims

CLAIMS1. A composition comprising: an anionic surfactant, an organic acid, and one or more compounds of Formula I:Formula I wherein R1is an unsaturated C3-C6 hydrocarbyl group, and R2is a C1-C3 alkyl group.

2. The composition according to claim 1, wherein the one or more compounds ofFormula I comprise or consist of one or both of and3. The composition according to claim 1 or 2, wherein the anionic surfactant comprises or consists of sodium dodecyl sulphate (SDS).

4. The composition according to any one of the preceding claims, wherein the organic acid comprises or consists of lactic acid.

5. The composition according to any one of the preceding claims, wherein the anionic surfactant is present in the composition in an amount of from 0.5 to 5 wt%, based on the total weight of the composition.

6. The composition according to any one of the preceding claims, wherein the organic acid is present in the composition in an amount of from 2.5 to 10 vol%, based on the total volume of the composition.

7. The composition according to any one of the preceding claims, wherein the one or more compounds of Formula I are present in the composition in an amount of from 0.015 to 1.0 wt%, based on the total weight of the composition.

8. The composition according to any one of the preceding claims, wherein the composition comprises an essential oil component which contains the one or more compounds of Formula I.

9. The composition according to claim 8, wherein the essential oil component comprises one or more of fennel oil, aniseed oil, basil oil, and tarragon oil.

10. The composition according to claim 8, wherein the essential oil component consists of fennel oil.

11. The composition according to any one of the preceding claims, wherein the composition comprises at least 0.5 wt% SDS, at least 2.5 wt% lactic acid and at least 0.05 wt% of an essential oil component comprising one or more of fennel oil, aniseed oil, basil oil, and tarragon oil.

12. The composition according to any one of the preceding claims, wherein the composition comprises from 1.0 to 2.0 wt% SDS, at least 4.5 wt% lactic acid and at least 1.0 wt% of an essential oil component comprising one or more of fennel oil, aniseed oil, basil oil, and tarragon oil.

13. A method of killing or preventing the growth of fungus, comprising contacting the fungus with the composition according to any one of claims 1-12.

14. The method according to claim 13, wherein the fungus comprises or consists of mould.

15. The method according to claim 13 or 14, wherein the method results in a mean logio reduction in the number of Aspergillus brasiliensis (ATCC 16404) spores per mL which is at least 2, optionally at least 4.

16. Use of a compound according to Formula I as an antifungal or fungicidal agent in a composition:Formula I wherein R1is an unsaturated C3-C6 hydrocarbyl group, and R2is a C1-C3 alkyl group.

17. The use according to claim 16, wherein the composition further comprises an anionic surfactant and an organic acid.

18. A method of manufacturing the composition according to any one of claims 1-12.

19. The method according to claim 18, wherein the method comprises: combining the anionic surfactant, the organic acid and the one or more compounds of Formula I; and mixing to form the composition.

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