An antimicrobial composition

[0001] The present invention relates to an antibacterial composition. In particular, it relates to an anti-bacterial composition intended for use in body washing, oral care or for cleaning hard surfaces.

[0002] Disinfecting and disinfecting soap compositions containing a chlorine-based antibacterial agent such as triclosan are known. To provide effective antibacterial action, such compositions require a rather long contact time. In practice, users, especially children, do not spend much time washing and, as a result, washing with such compositions does not provide adequate protection against surface or dermal infection, or adequate protection against disease. The user, despite washing his hands, is likely to have skin with rather inadequate bacterial removal and may contaminate further animate surfaces and / or dead surfaces and cause the spread of pathogens, and consequently diseases.

In general, users, and especially children, washing soiled hands before eating with a slow-acting antibacterial composition for a relatively short period of time risk contact with diseases. In addition, in addition to abrasives, many antibacterial active substances are incorporated into oral care compositions, such as compositions applied to teeth, but generally such active substances require several minutes, if not hours, before effective antibacterial action is obtained. People often brush their teeth or rinse their mouth for very short periods of time, for example, on the order of 1 minute or less, making such compositions quite ineffective in providing the desired benefits.

[0003] Similarly, in the area of ​​cleaning hard surfaces, e.g. cleaning floors, table surfaces or tools, the antibacterial substances contained in the composition are in contact with the ground for less than a few minutes, after which the surface is either wiped or rinsed with water. Such short periods of cleaning action are ineffective to provide the desired benefits because most known antibacterial substances commonly used in such products need several hours to provide the required bacterial killing.

[0004] Therefore, there is a need to provide a composition that gives a relatively more effective antibacterial effect when the cleaning / washing period is relatively short, usually about 5 minutes or less, preferably less than 2 minutes, and in many cases less than one minute. or sometimes as little as 15 seconds or less.

[0005] The inventors of the present invention have surprisingly found that compositions containing selected ingredients, namely thymol and terpineol, in selected proportions provide a relatively fast antibacterial effect.

[0006] GB-366 870 (Namlooz Vennootschap, 1931) describes the process of perfuming toilet soaps. Two different examples of soap perfumes are described, one containing thymol and the other containing terpineol. No disclosure of a specific soap composition comprising a mixture of thymol and terpineol.

[0007] US Patent Publication No. 6,534,042 (Pfizer, 2003) describes an oral care composition comprising from 0.01 to 5% by weight of a citrus aroma or citrus aroma component, and from 0.01 to 5% by weight of phenolic compounds selected from the group consisting of menthol, eucalyptol (cineol), methyl salicylate, thymol, triclosan, and mixtures thereof, and orally acceptable

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Carrier. The citrus aroma component is selected from the group consisting of limonene, citral, cadiene, decyl aldehyde, linalool, terpineol, linalyl esters, terpinyl acetate, citronellol, alpha-terpinene, 2-decanal, alpha-pinene, beta-pinene, 3-penternal, decanal , and C8 to C10 and C12 aldehydes, acids, and esters found in citrus aromas, and mixtures thereof. The oral care composition has been reported to be useful in retarding plaque development, in treating gums, and in reducing the population of viable oral microorganisms. No disclosure of a specific composition comprising thymol and terpineol.

[0008] GB-508 407 (Shepherd, 1938) describes an aseptic product and a method for its preparation comprising the steps of mixing salol and thymol in a 1: 3 weight ratio, melting this mixture and cooling to form crystals. The exemplary composition described contains 59 parts of crystals, 41 parts of terpineol, 200 parts of red turkey oil, and 200 parts of water. The composition described in this document contains about 8% by weight thymol and about 8% by weight terpineol, and is said to be particularly useful for disinfecting air. Similarly, the publication of international patent application WO 2008/088827 discloses several hundred embodiments of compositions intended for controlling insects and pests, one of which contains thymol and terpineol in similarly high concentrations.

[0009] US 2004/0014818 (Boeck) discloses a bactericidal formulation in the form of a solution, cream or ointment mixed with photosynthesized hydrocarbons, hydrocarbon isolates, 2-hydroxy-1-isopropyl-4-methylbenzene (thymol) and butylated hydroxytoluene, many such compositions are illustrated by examples, each having 10 to 20 compounds with antibacterial efficacy. This patent publication does not disclose that a particular combination of terpineol and thymol, in certain amounts, provides a rapid antibacterial effect.

[0010] None of the patents cited above relates to the problem of slow action of antibacterial compositions.

[0011] US Patent 5,965,518 (Nakatsu et al., 1999) describes a fragrance composition with antibacterial activity, containing between 3 and 20% by weight of a phenolic compound, and between 20 and 80% by weight of a non-aromatic terpenoid. Examples of phenolic compounds include amyl salicylate, cavacrol, dihydroeugenol, eugenol, hexyl eugenol, hexyl salicylate, isoeugenol, methyl eugenol, methyl isoeugenol, methyl salicylate, tert-butylcresol, thymol, and viline. Examples of non-aromatic terpenoid compounds include cedren, cineol, citral, citronellal, citronellol, cymene, paradihydrolinalool, dihydromyrcenol, farnesol, geraniol, hexyl cinnamic aldehyde, hydroxycytronallol, hydroxycitronellal, isocytral, limonene, linaloolol, menthol terpinenol, and tetrahydromyrcenol. There is no specific disclosure of a composition comprising thymol and terpineol. Such compositions reduce the bacterial titer by at least 1.5 log cfu / ml in 5 minutes when used at a concentration of about 0.25%. Although they provide some improvement in the speed of antibacterial activity, there remains a need for a composition with a relatively fast antibacterial effect. [0012] The object of the present invention is to avoid or alleviate at least one of the disadvantages of the prior art, or to provide a useful alternative.

[0013] Another object of the present invention is to provide antibacterial compositions that have a relatively fast antibacterial effect.

[0014] In accordance with a first aspect of the present invention, it provides an antibacterial composition comprising:

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(A) 0.01 to 5% by weight thymol, (b) 0.01 to 5% by weight terpineol, (c) 1 to 80% by weight of anionic surfactant, and (d) carrier.

[0015] According to a second aspect of the present invention, it provides a method for disinfecting a surface comprising the steps of:

(a) applying the composition of the first aspect of the present invention to a surface; and (b) flushing the surface with a suitable solvent.

[0016] These and other aspects, features and benefits will be apparent to those skilled in the art when reading the present detailed description and the appended claims. For the avoidance of any doubt, any features of one aspect of the present invention may be used in any other aspect of the invention. The word "contains" is intended to mean "embracing" but not necessarily "consists of" or "built of". In other words, the stages and options listed are not necessarily exhaustive. It is believed that the following examples are intended to illustrate the invention but not to limit the invention to examples only. Similarly, all percentages given are weight / weight percentages, unless otherwise stated.

[0017] With the exception of the described examples of the present invention and comparative examples, or unless explicitly stated otherwise, all numbers in this specification indicating the amounts of material or reaction conditions, physical properties of the materials, and / or use are to be understood as modified by the word " about". Unless otherwise specified, numerical ranges expressed as "from x to y" should be understood to include x and y values. If for a particular feature many preferred ranges are described in the form "from x to y", then it is understood that all ranges connecting different endpoints are also considered. [0018] The antibacterial composition comprises thymol, terpineol and a carrier. The various components of the antibacterial composition are described below. The compositions of the present invention are preferred for non-therapeutic use, and more specifically for use for cleaning the surface of the human body, including skin, hair or mouth, or for cleaning hard surfaces.

[0019] Preferably, the antibacterial composition contains 0.05 to 5%, more preferably 0.1 to 1%, and most preferably 0.1 to 0.4% by weight thymol. The most useful antibacterial compositions of the present invention have thymol more than 0.05 and less than 0.3% by weight thymol. Such preferred thymol concentration ranges are important because below the preferred low thymol concentration, the desired rapid action of antibacterial kinetics in combination with terpineol would not occur. At concentrations higher than higher preferred thymol concentrations, when present in combination with terpineol, where the kinetics of action would not be impaired, the present inventors have found that not as for therapeutic / pesticidal / herbicidal applications where sensory aspects are not critical, in the present application, which is preferably cleaning / washing the body, mouth, or use for cleaning hard surfaces, the product is in contact with the hands, lips or other body parts, and then sensory aspects such as smell and sensation on the skin would be worse. Thymol can be added to the antibacterial composition in purified form.

[0020] Alternatively, thyme oil or thyme extract containing thymol may be added to the antimicrobial composition, ensuring that thymol is present in the desired concentration in the composition of the present invention. Thyme oil or thyme extract is obtained from the thyme plant. The term "thyme" refers to plants belonging to the genus Thymus and includes, but is not limited

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EP 2 348 838 B1 to the following species: Thymus vulgaris, Thymus zygis, Thymus satureoides, Thymus mastichina, Thymus broussonetti, Thymus maroccanus, Thymus pallidus, Thymus algeriensis, Thymus serpyllum, Thymus pulegoide, and Thymus citriodorus.

[0021] The structure of thymol and its carvacrol isomer is as follows:

carvacrol

<img file="PL2348838T3_D0001.tif" />

thymol

<img file="PL2348838T3_D0002.tif" />

[0022] Preferably the antibacterial composition contains 0.05 to 5%, more preferably 0.1 to 1%, and most preferably 0.4 to 0.6% by weight terpineol. The most useful fast acting antibacterial compositions of the present invention have a terpineol content greater than 0.05 and less than 1% by weight. These preferred terpineol concentration ranges are important for some reasons because the thymol concentration ranges are favorable for satisfying the desired rapid action of antibacterial kinetics and are not sensory unpleasant when used in body care, oral care or hard cleaning products. surface. Preferably, terpineol is selected from alpha-terpineol, beta-terpineol, gamma-terpineol, or mixtures thereof. It is particularly preferred that terpineol is alpha-terpineol. Terpineol can be added to the antimicrobial composition in purified form.

[0023] Alternatively, terpineol-containing pine oil may be added to the antimicrobial composition.

[0024] The structure of the terpineol compound is shown below:

<img file="PL2348838T3_D0003.tif" />

[0025] The antibacterial composition comprises a carrier. The carrier is selected from the group consisting of water, oil, solvent, inorganic particulate material, starches, and mixtures thereof. Preferably, the carrier comprises from 0.1 to 99% by weight of the composition. The antibacterial composition may be in the form of a solid, liquid, gel, paste, or soft solid, and the carrier may be selected by a person skilled in the art depending on the format / form of the antibacterial composition chosen.

[0026] Examples of inorganic particulate materials include clay, talc, calcite, dolomite, silica, and aluminosilicate. Examples of oils include mineral oils, vegetable oils, and oils and waxes from the processing of petroleum. Examples of solvents include alcohols, ethers and acetone.

[0027] The starch may be natural starch obtained from food grains, or it may be a modified starch.

[0028] Particularly preferred carriers are water or an oil / solvent, and a more preferred carrier is a mixture of water and oil. In most anticipated uses, such as body care /

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For washing, oral care, and cleaning hard surfaces, the antibacterial composition may be formulated in an aqueous base (where the carrier is water), for example in gel products, or in a pure oil / solvent base, for example in anhydrous stick products or products containing a propellant. However, the most preferred product form has an emulsion base (and the carrier is water and oil), for example in liquid soap products, in the form of a lotion, or in the form of a semi-solid, intended for hand washing, face washing, for washing the body or for washing hair; for use in toothpastes / dental compositions intended for oral care, or in products designed to clean hard surfaces in the form of cubes or liquids. Thus, a particularly preferred antibacterial composition includes:

a. More than 0.05 and less than 0.3% by weight thymol;

b. More than 0.05 and less than 1% by weight of terpineol; and

c. carrier.

[0029] Preferably, the antibacterial composition contains 1 to 80% surfactant. Generally, the surfactants can be selected from the surfactants described in well-known textbooks such as "Surface Active Agents" volume 1 by Schwartz &amp; Perry, Interscience 1949, volume 2 by Schwartz, Perry &amp; Berch, Interscience 1958, and / either in the current edition of the publication "McCutcheon's Emulsifiers and Detergents" published by the Manufacturing Confectioners Company, or in the textbook "Tenside-Taschenbuch", H. Stache, 2nd edition, Carl Hauser Verlag, 1981. Any type of surfactant can be used, i.e. anionic, cationic, non-ionic, zwitterionic or amphoteric.

[0030] A particularly preferred surfactant is soap. Soap is a suitable surfactant for use in body anti-bacterial compositions of the present invention. Preferably, the soap is C8-C24 soap, more preferably C10-C20 soap, and most preferably C12-C16 soap. The soap may or may not have one or more carbon or carbon double or triple bonds. The cation in such soap may be an alkali metal, alkaline earth metal cation or an ammonium cation. Preferably, the cation in the soap is selected from sodium, potassium or ammonium. More preferably the cation in the soap is sodium or potassium.

[0031] Soap can be made by saponifying fat and / or fatty acid. Generally, the fats and oils used for making soap may be such as tallow fat, tallow stearin, palm oil, palm stearin, soybean oil, fish oil, castor oil, rice bran oil, sunflower oil, coke oil, babassu oil , palm kernel oil, and more. In the above process, the fatty acids are derived from oils / fats selected from those derived from coconut, rice bran, peanuts, tallow, palm, palm kernels, cotton seeds, soybeans, castor oil, etc. Fatty acid soaps can also be made synthetically (e.g. by oxidation of petroleum, or by hydrogenation of carbon monoxide in the Fischer-Tropsch process). Resin acids such as those found in tallow oil may be used. Naphthenic acids are also suitable.

[0032] Tallow fatty acids may be derived from various animal sources, and generally contain about 1-8% myristic acid, about 21-32% palmitic acid, about 14-31% stearic acid, about 0-4% palmitoleic acid, about 36-50% oleic acid, and about 0-5% linolenic acid. Typical distribution is 2.5% myristic acid, 29% palmitic acid, 23% stearic acid5

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EP 2 348 838 B1, 2% palmitoleic acid, 41.5% oleic acid, and 3% linolenic acid. Other similar mixtures, such as those derived from various animal tallow and lard, are also suitable.

[0033] Coconut oil refers to mixtures of fatty acids having an approximately carbon chain length distribution of 8% C8, 7% C10, 48% C12, 17% C14, 8% C16, 2% C18, 7% oleic acid and 2 % linolenic acid (the first six of these fatty acids are saturated). Other sources that have similar carbon chain length distributions, such as palm kernel oil and babassu kernel oil, are covered by the term coconut oil.

[0034] A typical fatty acid blend consists of 5 to 30% coconut fatty acids, and 70 to 95% fatty acids derived from hardened rice bran oil. Fatty acids derived from other suitable oils / fats, such as peanuts, soybeans, tallow, palm, palm kernels, etc. may also be used in other appropriate proportions. The soap, when present in a solid form in accordance with the present invention, is present in an amount of 30 to 90%, preferably 50 to 85%, and more preferably 55 to 75% by weight of the composition. Soap, when present in a liquid form composition, is present in an amount of 0.5 to 20%, preferably 1 to 10% by weight of the composition.

[0035] The antibacterial composition of the present invention is useful in applications for cleaning hard surfaces. Preferred surfactants for such applications are non-ionic surfactants, such as ethoxylated C8-C22 and preferably C8-C16 fatty alcohols, containing between 1 and 8 ethylene oxide groups when the product is in liquid form. When the product is a solid product intended for use in the cleaning of hard surfaces, preferably the surfactants are selected from primary alkyl sulfates, secondary alkyl sulfonates, alkyl benzene sulfonates, or ethoxylated alkyl sulfates. The composition may still contain an anionic surfactant, such as an alkyl ether sulfate, preferably one that has between 1 and 3 ethylene oxide groups, either from a natural or synthetic source, and / or sulfonic acid. Sodium lauryl ether sulfates are particularly preferred. Alkyl polyglucosides may also be present in the composition, preferably those having a carbon chain length between C6 and C16. Suitable concentrations of surfactant in applications for cleaning hard surfaces in liquid form are generally from about 0.5 to 10%, and preferably from 1 to 5% by weight of the composition. In solid compositions, the surfactant is preferably present in amounts of 5 to 40%, preferably 10 to 30% by weight of the composition.

[0036] The antibacterial composition of the present invention is useful in oral care compositions, for example in dental / toothpaste compositions, or a mouthwash product. In such applications, preferred surfactants are anionic, nonionic or amphoteric surfactants, preferably anionic or amphoteric. Preferably, the anionic surfactant is an alkali metal alkyl sulfate, and more preferably sodium lauryl sulfate (SLS). Mixtures of anionic surfactants may also be used. A preferred amphoteric surfactant is betaine, and more preferably alkylamidopropyl betaine (in which the alkyl group has a linear chain C10-C18), and most preferably it is cocamidopropyl betaine (CAPB). Mixtures of amphoteric surfactants may also be used. In general, appropriate surfactant concentrations in care applications

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% Of the oral cavity is from about 2% to about 15%, preferably from about 2.2% to about 10%, and more preferably from about 2.5 to about 5% by weight of the total composition.

[0037] Thus, in a very preferred aspect of the invention, the antimicrobial compositions contain soap, alkyl sulfate or (linear alkyl) benzenesulfonate as surfactants.

[0038] The inventors of the present invention quite unexpectedly determined that when thymol alone and terpineol alone alone do not provide rapid antimicrobial kinetic activity, then the combination of thymol and terpineol at specific concentrations provides a synergistic antibacterial effect that is particularly important in washing operations when contact of antibacterial active substances with the surface is short, i.e. it is on the order of less than 5 minutes, and preferably less than 2 minutes, even more preferably less than a minute, and in many cases less than 15 seconds. Preferably, such washing processes involve the use of a surfactant for cleaning action. Even more surprised by the inventors, the surfactant alone does not provide rapid killing of microbes at the concentration found in washing processes, but it provides further improvement in reducing bacterial titer on the surface in a short period of time when the surface is washed with a composition comprising terpineol, thymol and additional surfactant. Thus, while on the one hand the surfactant is generally known to be responsible for cleaning the dirt as well as the antibacterial active substances used in the composition, in the present invention it provides highly useful additional benefits in that it increases the reduction of bacterial titer in a composition containing a combination of thymol and terpineol only.

[0039] Further additional advantages of the present invention are that it is observed that surfaces treated with a composition comprising terpineol and thymol, surprisingly, allow continuous surface protection against bacterial growth for a significant period of time after application.

[0040] The composition may still contain various additional ingredients known to those skilled in the art. Such additional ingredients include, but are not limited to: fragrances, pigments, preservatives, emollients, sunscreen protectors, emulsifiers, gelling agents or thickeners, propanol, are particularly preferred as a carrier for the antibacterial cloth / washer, or an antibacterial composition intended for decontaminating the hands.

[0041] In accordance with another aspect of the present invention, it provides a method of disinfecting a surface, the method comprising the steps of:

(a) applying the composition of the first aspect of the present invention to a surface; and (b) flushing such surface with a suitable solvent.

[0042] A preferred solvent for flushing the surface is water, but it could also be a mixture of water and alcohol. The word "flushing" as used herein includes the act of wiping the surface with a suitable cloth / washcloth. Thus, such a surface, e.g., hand, face, body, mouth, or any hard surface, e.g. equipment, is first contacted with the composition of the present invention. It is then preferably rinsed with sufficient water after a predetermined period of time to remove any visible or felt residual composition. Alternatively, the alcohol wipe / cloth or the water / alcohol impregnated cloth / cloth may be used to wipe the surface so that it is visibly free from the antibacterial composition. Preferably, the substrate rinsing step is carried out in less than 5 minutes, preferably

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In less than 2 minutes, and even more preferably less than a minute, and in many cases less than 15 seconds, after the step of applying the composition to the given substrate.

[0043] In accordance with another aspect of the invention, it provides non-therapeutic benefits.

[0044] Thus, in accordance with yet another aspect of the invention, it provides the use of a composition comprising 0.01 to 5% by weight thymol, 0.01 to 5% by weight terpineol, and a carrier for faster reduction of bacterial titer.

[0045] According to yet another aspect of the invention, it provides the use of a composition comprising 0.01 to 5% by weight thymol, 0.01 to 5% by weight terpineol, and a carrier, for improving surface hygiene of the human body. Such human surfaces include skin, hands and mouth. In a preferred aspect, there is provided the use of a composition comprising 0.01 to 5% by weight thymol, 0.01 to 5% by weight terpineol, and a carrier, for improved hand hygiene. Yet another preferred aspect of the invention provides a composition comprising 0.01 to 5% by weight thymol, 0.01 to 5% by weight terpineol, and a carrier for use in improved oral hygiene.

[0046] In accordance with another aspect of the invention, also the inorganic particulate material is a suitable carrier. When such an inorganic particulate material is a carrier, the antibacterial composition is in solid form. Talc is the preferred inorganic particulate material. When talc is the inorganic particulate material, the solid antibacterial composition is particularly useful as talc powder for face or body.

[0047] According to a further aspect, the preferred carrier is a solvent. Although any solvent may be used, the preferred solvent is alcohol. Short chain alcohols, in particular ethanol and propanol, are particularly preferred as a carrier for an antibacterial cloth / washer, or for an antimicrobial composition intended for hand sanitizing.

[0048] In accordance with another aspect of the present invention, it provides a method of disinfecting a surface, the method comprising the steps of:

(a) applying the composition of the first aspect of the present invention to a surface; and (b) flushing such surface with a suitable solvent.

[0049] Preferably the solvent for flushing the surface is water, but also it can be a mixture of water and alcohol. The word "flushing" as used herein includes wiping / rubbing the surface with a suitable cloth. Thus, the surface, e.g. of the hand, face, body, mouth, or any hard surface, e.g. of equipment, is first contacted with the composition of the present invention. It is then preferably rinsed with sufficient water after a predetermined period of time to remove any visible, residual composition residues. Alternatively, an alcohol cloth / washcloth may be used, or a water / alcohol impregnated cloth / washcloth may be used to wipe the surface so that it is visibly free of the antibacterial composition. Preferably, the rinsing step of the substrate is carried out for less than 5 minutes, more preferably less than 2 minutes, and more preferably less than a minute, and in many cases less than 15 seconds, after the step of applying the composition to the given substrate.

[0050] In accordance with another aspect, the present invention provides non-therapeutic benefits.

[0051] Thus, in accordance with yet another aspect of the present invention, it provides the use of a composition comprising 0.01 to 5% by weight thymol, 0.01 to 5% by weight terpineol, and a carrier for faster reduction of bacterial titer.

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[0052] According to yet another aspect of the invention, it provides the use of a composition comprising 0.01 to 5% by weight thymol, 0.01 to 5% by weight terpineol, and a carrier, for improving surface hygiene of the human body. Such human surfaces include skin, hands and mouth. In a preferred aspect, there is provided the use of a composition comprising 0.01 to 5% by weight thymol, 0.01 to 5% by weight terpineol, and a carrier, for improved hand hygiene. In yet another aspect, it provides the use of a composition comprising 0.01 to 5% by weight thymol, 0.01 to 5% by weight terpineol, and a carrier, for improving oral hygiene.

[0053] The invention also provides therapeutic benefits.

[0054] Thus, according to another aspect of the invention, it provides a composition comprising 0.01 to 5% by weight thymol, 0.01 to 5% by weight terpineol, and a carrier for faster reduction of bacterial titer.

[0055] In accordance with another aspect of the present invention, it provides the use of a composition comprising 0.01 to 5% by weight thymol, 0.01 to 5% by weight terpineol, and a carrier for improving surface hygiene of the human body.

Examples [0056] The invention will now be illustrated by way of examples. The examples are given for illustration only and do not limit the scope of the claims in any way.

Examples 1 to 3:

Synergistic interaction between thymol and terpineol in providing bacterial killing in a short time (15 seconds) [0057] Compositions were prepared containing each single active substance and their combination, with water as carrier (detailed composition is given in Table 1 below). Terpineol and thymol were obtained from Nishant aromas, India.

[0058] About 10 were taken into the test tube⁷ bacterial cells (E. coli ATCC 10536) and contacted with various compositions for a period of 15 seconds. After 15 seconds of contact, the bacteria were removed and the presence of live bacteria was determined by serial dilution and placing on agar plates. The results were presented in the form of log (live E. Coli) value which is log10 from the number of live E. Coli bacteria remaining after 15 seconds of contact. So if there are 10 left⁴, then the log (live E. Coli) is 4.

Table 1: Antibacterial efficacy of thymol and terpineol

<td>Example Number</td><td>Composition</td><td>Log (live E. coli)</td>

<td> 1</td><td>thymol (0.2%)</td><td> 3,1</td>

<td> 2</td><td>terpineol (0.5%)</td><td> 6,3</td>

<td> 3</td><td>thymol (0.2%) + terpineol (0.5%)</td><td>There were no live bacteria left</td>

[0059] The results shown in Table 1 show that there is a synergistic interaction between thymol and terpineol in providing bacterial killing in a short period of time.

Examples 4 to 9:

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Efficacy of other antibacterial combinations in killing bacteria in a short period of time (15 seconds) [0060] Experiments similar to those described in Examples 1 to 3 were performed with various other antibacterial combinations as shown in Table 2 below. Eucalyptol and linalool were obtained from Fluka. Menthol, citral, and geraniol were obtained from Som Santi Corporation, India. At first 10 were downloaded⁸bacterial cells (E. coli ATCC 10536). Results are expressed in log values ​​(live E. Coli) for various combinations after 15 seconds of contact, and are shown in Table 2 below.

Table 2: Antibacterial efficacy

<td>Example Number</td><td>Composition</td><td>Log (live E. coli)</td>

<td> 4</td><td>thymol (0.2%) + linalool (0.5%)</td><td> 3,4</td>

<td> 5</td><td>thymol (0.2%) + citral (0.5%)</td><td> 5,0</td>

<td> 6</td><td>thymol (0.2%) + geraniol (0.5%)</td><td> 4,1</td>

<td> 7</td><td>menthol (0.2%) + terpineol (0.5%)</td><td> 5,0</td>

<td> 8</td><td>eucalyptol (0.2%) + terpineol (0.5%)</td><td> 2,1</td>

<td> 9</td><td>triclosan (0.2%) + terpineol (0.5%)</td><td> 5,0</td>

[0061] The results shown in Table 2 show that various other combinations of well-known compounds of a similar class as thymol and terpineol do not provide rapid antibacterial activity.

Examples 10 to 13:

Efficacy of thymol and terpineol combinations in inhibiting the growth of various other bacteria compared to TCC.

[0062] The combination of thymol (0.2%) and terpineol (0.5%) was tested for effectiveness in inhibiting the growth of various bacteria after contacting this combination for about 15 seconds. Similar experiments were performed with another well-known substance with antibacterial activity of TCC (trichlorocarbanilide). The experiment was conducted in a similar manner to that of Examples 1 to 3, and the results for the various bacteria remaining after contact with the active substance for 15 seconds are summarized in Table 3 below

Table 3: Antibacterial efficacy of active substances for various bacteria

<td>Example No.</td><td>Organism</td><td>Kennel added Log (CFU / ml)</td><td>0.2% TCC Log (CFU / ml)</td><td>0.2% thymol + 0.5% terpineol Log (CFU / ml)</td>

<td> 10</td><td>Klebsiella</td><td> 7,35</td><td> 7,28</td><td> 3,4</td>

<td> 11</td><td>Entereobacter</td><td> 6,87</td><td> 6,87</td><td> 5,0</td>

<td> 12</td><td>Salmonella</td><td> 7,63</td><td> 7,33</td><td> 4,1</td>

<td> 13</td><td>Vibrio cholerae</td><td> 7,58</td><td> 7,20</td><td> 5,0</td>

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[0063] The results shown in Table 3 show that the composition of the present invention is active against the large number of bacteria encountered when cleaning various substrates, while the well-known antibacterial substance (TCC) does not provide such efficacy.

Examples 14-17:

Antibacterial efficacy of soaps containing various antibacterial active substances [0064] Soap bars (containing 72 TFM soaps) containing various antibacterial active substances were prepared. Such soap bars were tested in in-vitro tests according to the procedures used in Examples 1 - 3. The sample used for analysis was 8% aqueous soap bar solution. The amount of E-Cola bacteria initially added was about 10⁷. Various soap bars were made and the results of antibacterial efficacy in terms of the amount of residual live bacteria after contact with the active substance for 15 seconds are summarized in Table 4 below.

Table 4

<td>Example Number</td><td>Composition</td><td>Log (CFU / ml)</td>

<td> 14</td><td>Soap</td><td> 6,35</td>

<td> 15</td><td>Soap with 0.2% carvacrol</td><td> 6,02</td>

<td> 16</td><td>Soap with 0.2% carvacrol + 0.5% terpineol</td><td> 6,78</td>

<td> 17</td><td>Soap with 0.2% thymol + 0.5% terpineol</td><td> 5,21</td>

[0065] Carvacrol is the isomeric form of thymol. The results shown in Table 4 show that the soap bar made with the composition of the present invention (Example 17) is better than soap bars made without any antibacterial substance or with other antibacterial substances, although some of these active substances have a thymol-like structure.

Examples 18 to 21:

Antibacterial activity of the soap composition when washing hands artificially contaminated with bacteria [0066] The following protocol was used to test the antibacterial efficacy of various compositions in their ability to remove bacteria applied to human hands.

[0067] Both hands of the volunteers were first disinfected to remove normal flora from the hands. 100 microliters of E.coli suspension (equivalent to about 10⁷ CFU / ml) applied to both hands of volunteers. One hand was washed with a soap solution containing 8% soap, but without the antibacterial active substance in the soap solution. The other hand was washed with soap containing one or more antibacterial active substances. Hands washed for 15 seconds on tester.

[0068] Next, the following steps were performed to obtain bacteria remaining on the hands of volunteers. The hand was placed in a sterile polyethylene bag containing 75 ml bacterial collection fluid (Butterfield phosphate buffer with neutralizers), and secured above the wrist with a rubber band. Bacteria collection was carried out using an automatic hand washing device in which the polyethylene bag was scrubbed on the outside with mechanical brushes for earlier

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A certain period of time during which most bacteria was removed from the palm of the hand and dispersed in the bacterial collection fluid. Samples were immediately diluted and placed on CY agar plates. Antibacterial efficacy in terms of the amount of remaining live bacteria on the hand after washing with the soap composition for 15 seconds is summarized in Table 5 below.

Table 5

<td>Example</td><td>Composition</td><td>Log (remaining live E, cola)</td>

<td> 18</td><td>Soap</td><td> 4,00</td>

<td>18A</td><td>Soap with 0.2% thymol</td><td> 3,51</td>

<td> 19</td><td>Soap</td><td> 4,06</td>

<td>19A</td><td>Soap with 0.5% terpineol</td><td> 4,10</td>

<td> 20</td><td>Soap</td><td> 3,30</td>

<td>20A</td><td>Soap with 0.2% carvacrol and 0.5% terpineol</td><td> 3,43</td>

<td> 21</td><td>Soap</td><td> 5,23</td>

<td>21A</td><td>Soap with 0.2% thymol and 0.5% terpineol</td><td> 4,32</td>

[0069] The results shown in Table 5 show that the soap composition of the present invention (Example 21A) provides significantly better antibacterial efficacy compared to a soap composition containing neither thymol nor terpineol (Example 21). For comparison, no better benefit was seen for the soap containing the antibacterial substance compared to the respective control samples (compositions of Examples 18A, 19A and 20 compared to the compositions of Examples 18, 19, and 20).

Examples 22 and 23:

Extended protection [0070] The ability of the compositions of the present invention to provide extended / further / protection on hands washed with the compositions of the present invention was tested. The protocol used is shown below.

[0071] One hand of the volunteer was washed with 8% 72 TFM soap solution (not containing an antibacterial active substance). The other hand was washed with a similar soap, but which additionally contained 0.5% terpineol and 0.2% thymol, for 15 seconds, per tester. Hands were allowed to air dry.

[0072] On both inner palm surfaces, three areas were marked, each with an area of ​​~ 2.8 ₂ cm². At a time of 0 minutes, 30 minutes and 60 minutes, 25 microliters corresponding to approximately 10 were applied to each area⁶ E.coli cells. After 5 minutes, the bacteria were harvested using a Teflon crucible and rod, and the bacteria were counted using CY agar. Antibacterial efficacy in terms of the amount of remaining live bacteria on the palm of the hand after the above procedure is summarized in Table 6 below.

Table 6

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EP 2 348 838 B1

<td>Example</td><td>Composition</td><td colspan="3">Log (other live E. coli)</td>

<td></td><td></td><td>0 minutes</td><td>30 minutes</td><td>60 minutes</td>

<td> 22</td><td>Soap</td><td> 5,70</td><td> 5,67</td><td> 5,25</td>

<td> 23</td><td>Soap with 0.2% thymol and 0.5% terpineol</td><td> 1,48</td><td> 1,96</td><td> 1,69</td>

[0073] The results shown in Table 6 show that the soap composition according to the present invention (Example 23) provides extended palm protection compared to soap compositions outside the scope of the invention.

Examples 24 - 29:

Antibacterial activity of the compositions of the present invention against oral bacteria [0074] The antibacterial activity of the compositions of the present invention was tested against commonly occurring gram positive oral bacteria, namely Streptococcus mutans ATCC 25175, and gram negative oral bacteria Neisseria subflava ATCC 19243). This was tested using oxoplates. Oxo-platelets monitor oxygen consumption in the reaction system (containing bacteria, wort and active ingredient).

Bacteria [0075] Gram positive (Streptococcus mutans ATCC 25175) and gram negative (Neisseria subflava ATCC 19243) were selected for testing active solutions. Both bacteria were cultured in BHIS agar plates for 24 hours at 37 ° C / 15% CO2, then suspended in PBS according to the McFarland 2 standard (~ 6x10⁸ cells).

Solutions [0076] For testing, each active substance was dissolved in 50% ethanol solutions. All solutions were prepared as 10 fold condensed, which allowed them to be diluted on the plate. Lower concentrations were used for N. subflav due to the greater sensitivity of this organism to biocides,

The oxblade method [0077] The following components were placed in the oxblade wells,

170al BHI wort 20al test solution 10O bacterium [0078] First, the wort was placed on the oxyplate, then the test solution, then the bacterium. Water or 50% ethanol was also used for each plate as controls. All solutions were tested in duplicate. Plates were placed on a fluorescent plate reader, incubated at 37 ° C and measured every 15 minutes for 18 hours. Sensors at the bottom of the wells were measured at two wavelengths, indicator (650/540 nM) reference (590/540 nm).

[0079] Using these values, the oxygen concentration was calculated and calibrated for maximum and minimum oxygen concentration. Then, a graph of the oxygen consumption curve over 18 hours that was used for was plotted

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Determining the level at which the bacterium begins to consume oxygen. Table 7 below shows the concentration of active substance at which no oxygen consumption was observed to indicate that there was no bacterial growth.

Table 7

<td>Example Number</td><td>Bacteria</td><td>Active substance concentration (% by weight) in the absence of bacterial growth</td>

<td> 24</td><td>Streptococcus mutans</td><td>Terpineol (0.4%)</td>

<td> 25</td><td>Streptococcus mutans</td><td>Thymol (0.08%)</td>

<td> 26</td><td>Streptococcus mutans</td><td>Terpineol (0.1%) + Thymol (0.04%)</td>

<td> 27</td><td>Neisseria subflava</td><td>Terpineol (0.08%)</td>

<td> 28</td><td>Neisseria subflava</td><td>Thymol (0.02%)</td>

<td> 29</td><td>Neisseria subflava</td><td>Terpineol (0.03%) + Thymol (0.012%)</td>

[0080] The results presented in Table 7 show that there is a synergistic interaction between thymol and terpineol in inhibiting the growth of oral bacteria.

[0081] The minimum concentration needed to inhibit the development of Streptococcus mutans in the above examples was 0.1% terpineol in combination with 0.04% thymol, and the concentration needed to inhibit the development of Neisseria subflava was 0.03% terpineol (0.03%) in combination with 0.012% thymol. However , it is preferred that the concentration needed to inhibit the growth of oral microbes in a short time be in the range of 0.05 to 5% by weight thymol in combination with 0.05 to 5% by weight terpineol.

Examples 30 - 33:

Soap compositions made with different amounts of thymol and terpineol.

[0082] Various soap bars (72 TFM) with an antibacterial active substance were prepared, as shown in Table 8 below. The bars were evaluated for the acceptability of the perceived odor assessed by a team of trained fragrance experts. The results are shown in Table 8 below.

Table 8

<td>Example Number</td><td>Composition</td><td>Odor evaluation</td>

<td> 30</td><td>Soap with 0.2% thymol and 0.5% terpineol</td><td>Highly accepted</td>

<td> 31</td><td>Soap with 0.4% thymol and 1.0% terpineol</td><td>Acceptable, but less favorable</td>

<td> 32</td><td>Soap with 0.8% thymol and 2% terpineol</td><td>Acceptable, but less favorable</td>

<td> 33</td><td>Soap with 8% thymol and 8% terpineol</td><td>Unacceptable</td>

[0083] From the results of all the above examples, it can be clearly seen that the particular mixture of terpineol and thymol of the present invention provides fast antibacterial activity with relatively high antibacterial efficacy compared to prior art compositions. The results still show

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And that a mixture of terpineol and thymol provides a synergistic rapid antibacterial effect with relatively high antibacterial efficacy compared to terpineol alone or thymol alone.