WO2015124943A1

WO2015124943A1 - Cleaning composition

Info

Publication number: WO2015124943A1
Application number: PCT/GB2015/050498
Authority: WO
Inventors: Sheika Fatima AL THANI
Original assignee: HART, Deborah Mary
Priority date: 2014-02-20
Filing date: 2015-02-20
Publication date: 2015-08-27
Also published as: GB201403017D0; GB2523340A

Abstract

The invention relates to a cleaning composition, in particular a sanitising composition. The composition disclosed can be used to sanitise body cleaning articles. In addition the compositions of the invention can be used to sanitise surfaces.

Description

CLEANING COMPOSITION

There is a general desire to keep the environments where we live clean and free from microorganisms, in particular those which may be harmful to our health. The cleaning products generally used contain chemicals which are harmful or cause irritation when touched with bare skin. There is a need for compositions which can remove microorganisms, but which are not harmful to human skin.

Surfaces, such as kitchen worktops, hobs, and sanitary ware, are exposed to microorganisms, in particular those which can cause sickness in people. The presence of microorganisms on these surfaces represents a risk to health, especially vulnerable people such as the elderly, children and immunocompromised patients. There is a need to provide cleaning compositions which can effectively clean these surfaces by removing dirt and grease, and leave a sanitising layer on the surface. Body cleaning articles are frequently used to cleanse and exfoliate the skin for example during a shower or bath. After use the materials are squeezed dry and/or simply allowed to air dry. A large amount of moisture is retained in the articles after use. In addition they are often kept in bathrooms which have a humid environment. This allows microorganisms removed from the skin and present in the environment to grow and multiply on the articles. In addition, over time the articles can become dirty or discoloured during use. Whilst some articles can be washed, others are often discarded as they cannot be easily cleaned, or become damaged during the cleaning process. There is a need to provide compositions that can be used to clean and/or sanitize body cleaning articles. The invention provides a cleaning composition comprising an antimicrobial agent dissolved in a mixture of an alcohol and an aqueous medium. The composition preferably comprises 2- 75wt% alcohol, 0.5-5wt% antimicrobial agent and 24-97wt% aqueous media. A "cleaning" composition is one that removes dirt from the article being cleaned. Preferably the composition is a sanitising composition. As used herein "sanitise" or "sanitising" means to reduce the number of microorganisms and optionally to remove dirt. The composition of the invention comprises an antimicrobial agent, preferably a natural antimicrobial agent. As used herein an "antimicrobial" agent is one that reduces the number of living microorganisms present. Such microorganisms are any one or more of bacteria, fungi, algae and viruses. The composition comprises an effective amount of the antimicrobial agent, i.e. at a concentration high enough to reduce the number of microorganisms present by 50wt%, 60wt%, 70wt%, 75wt%, 80wt%, 90wt%, 95wt%, or 99wt%. The antimicrobial agent may be present at a concentration of 0.5-25wt%, preferably l-10wt%, more preferably 2-5wt% of the total composition. The antimicrobial agent may be present at a concentration of 250 - 20,000 ppm, 500- 10,000ppm; 625- 5,000ppm; 1,250 -2,500ppm. The antimicrobial agent may be present at a concentration of at least 250ppm, 500ppm, 650ppm, 1250ppm, 2500ppm, 5000ppm or

10,000ppm.

Preferred antimicrobial agents include plant derived essential oils. Such oils are well known to the person skilled in the art and include, for example, tea tree oil, rose geranium oil and Fragonia oil. Tea tree oil, or melaleuca oil, is obtained from the leaves of the Melaleuca alternifolia. It is known to be an effective topical antimicrobial agent active against a wide range of

organisms. It has been reported that Tea tree oil may have a clinical application in both the hospital and community, especially for clearance of methicillin-resistant Staphylococcus aureus (M SA) carriage or as a hand disinfectant to prevent cross-infection with Gram- positive and Gram-negative epidemic organisms. The study, based on the time-kill approach, determined the kill rate of tea tree oil against several multidrug-resistant organisms,

including MRSA, glycopeptide-resistant enterococci, aminoglycoside-resistant Klebsiellae, Pseudomonas aeruginosa and Stenotrophomonas maltophilia, and also against sensitive microorganisms. The study was performed with two chemically different tea tree oils. One was a standard oil and the other was Clone 88 extracted from a specially bred tree, which has been selected and bred for increased activity and decreased skin irritation. The results confirm that the cloned oil had increased antimicrobial activity when compared with the standard oil. Most results indicated that the susceptibility pattern and Gram reaction of the organism did not influence the kill rate. A rapid killing time (less than 60 min) was achieved with both tea tree oils with most isolates, but MRSA was killed more slowly than other organisms. Preferably the tea tree oil is derived from a Clone 88 Tree. Rose geranium essential oil is extracted from the plant Pelargonium graveolens. Rose

geranium oil is a known antiseptic, and has a number of other beneficial properties such as antidepressant, astringent, promoting wound healing, diuretic, and deodorant.

Fragonia oil is obtained by steam distillation from the lignotubureous shrub, Agonis

Fragrans. it has been found to have a number of beneficial properties. It acts as an antiinflammatory agent, a decongestant agent by breaking up mucus, soothes the irritation and nervous cough reactions linked to the lungs, an analgesic when used externally, an

antimicrobial, and in regulating hormonal imbalances during the female menstrual cycle. It also helps to fights most common respiratory infections from the nose through to the lungs and is thought to strengthen and balance the immune system. Studies at the University of Western Australia have shown Fragonia to have significant anti-microbial activity, similar to that of other recognised anti-microbial essential oils. It has also been reported that Fragonia has the ability to balance, regulate, calm and help people let go of negative patterns of behaviour. It helps induce a deep and holistic healing process. It is also thought to be

valuable in dealing with unresolved core issues and transforming troubled family

relationships. In a preferred embodiment the composition comprises Tea Tree oil and Fragonia oil. It has been surprisingly found that Tea Tree oil and Fragonia oil have a synergistic antimicrobial effect when used together. The Tea Tree oil and Fragonia oil may each be present at a concentration of 250 - 20,000 ppm, 500-10,000ppm; 625- 5,000ppm; 1,250 -2,500ppm. The Tea Tree oil and Fragonia oil may each be present at a concentration of at least 250ppm, 500ppm, 650ppm, 1250ppm, 2500ppm, 5000ppm or 10,000ppm. Preferably the composition comprises at least 1250ppm Tea Tree oil and at least 1250ppm Fragonia oil.

The composition comprises an alcohol. Preferred alcohols include ethanol, denatured alcohol or isopropanol. According to the Centers for Disease Control and Prevention, ethanol-based hand antiseptics containing 60 to 90 percent ethanol appear to be most effective against common pathogens. Ethanol-based sanitizers also seem to have greater antimicrobial effects against viruses than isopropanol-based sanitizers. The composition may comprise l-75wt% alcohol, preferably 10-70wt% alcohol, more preferably 20-60wt%, 25-50wt% or 30-40wt% alcohol. The composition may comprise less than 40wt%, 30wt%, 20wt%, 15wt% or 10wt% alcohol. The composition may contain more alcohol than aqueous media, in which case it is designated an alcohol-based composition. An alcohol-based composition comprises about 55-70wt% alcohol, preferably about 60-67wt%, more preferably about 62-65wt% of the total composition. Preferably the composition comprises about 66wt% alcohol.

The composition may contain more aqueous media than alcohol, in which case it is designated a water-based composition. Water-based compositions comprise about 55-80wt% aqueous media, preferably about 60-75wt%, more preferably about 62-70wt% of the total composition. Preferably about 75wt% of the total water-based compositions is aqueous media.

As used herein "aqueous media" is defined as water or one or more plant extracts mixed with water. The water used in the compositions is preferably deionized water. Suitable plant extracts are well known to the skilled person. The plant extract water may be derived in a number of ways including: using undiluted extract from the plant mixed with water; dilution of a concentrated plant extract in water; and mixing a dried extract, for example a freeze dried extract, with water. Preferably the concentration of extract used is the equivalent of at least 5g/L dried extract, for example 5-20 g/L, preferably 10-15g/L. In a preferred embodiment the aqueous media is aloe vera water produced by mixing aloe vera extract with water. The aloe vera extract used can be derived from any part of the plant, preferably from the inner leaves of the plants, using methods known to the skilled person. The composition may comprise 2-97wt% aqueous media. The composition may comprise 10-95wt%, 20-90wt%, 25-85wt%, 30-80wt%, 35-75wt%, 40-70wt%, 45-65wt% or 50-55wt% aqueous media. In one embodiment the composition comprises 24- 97wt% aqueous media. Preferably the composition of the invention comprises natural ingredients. "Natural" as used herein means that the ingredient is produced within a living organism. Preferably all of the active ingredients in the composition are natural. Even more preferably all of the ingredients in the composition are natural. Preferred natural ingredients are derived from plants. However, the compositions of the invention can also be formulated using non-plant derived or synthetic ingredients, including synthetic analogues of the natural ingredients.

The composition of the invention can be used to sanitise body cleaning articles. Body Cleaning articles are used to apply soap or other cleansing products such as body wash or shower gel to the body including the face whilst washing for example in the bath or shower. Alternatively, or in addition, these articles are used to exfoliate the skin on the body. Examples of these articles include flannels, body mitts, loofahs, body brushes, sponges, shower puffs or scrubs. Body cleaning article may be natural, such as sponges or loofahs, or may be synthetic, such a body mitts and shower puffs. The composition of the invention can be left to air dry on the cleaning article or surface. It does not need to be washed off. A sanitising layer is left on the article or surface. Body cleaning articles may be used on the skin of people of all ages, including children, after application of the composition. In addition the surfaces cleaned by the composition may be touched. Therefore the composition should be gentle on the skin of users, and not cause adverse reactions such as irritation and dryness.

The presence of agents beneficial to the skin in the composition is preferred. In addition it is desirable for the composition to have a pleasant perfume. The composition can also be coloured so that it is visually attractive before and during use. Thus the composition may further comprise one or more additives, which are preferably natural. Suitable additives include, for example, perfumes, colouring agents and essential oils.

Colouring agents may be natural dyes or a synthetic dye, for example a polymeric colouring agent. Polymeric colouring agents, such as Liquitint^®, are non-staining, and more stable than natural dyes. They generally do not fade. In addition, as they are not absorbed by the skin they are are considered safer than normal food grade dyes. The dye is used in an amount necessary to obtain the desired depth of colour.

Essential oils can be added to impart fragrance and/or enhance sanitising properties. Examples of essential oils which enhance sanitisation are lavender oil, lemon, neroli, peppermint or ylang ylang. Other essential oils can be included for fragrance only. The preferred concentration of essential oils in the composition is about 0.01 to 0.5g/L, preferably 0.1 to 0.25g/L.

The composition may further comprise one or more excipients, which are preferably natural. Suitable excipients are well known to the skilled person include, as examples, emollients, surfactants, stabilisers, foaming agents, preservatives and humectants.

Emollients soften the skin and can soothe irritated skin. Examples of suitable emollients include vegetable, seed and nut oils, aloe vera extract, and glycerol. Emollients may be present in an amount of about 0.5-20wt%, more preferably about 1- 10 wt%, most preferably about 3-8wt% of the total composition.

Surfactants are compounds that lower the surface tension between two liquids. In the composition of the invention they can be used to help solubilise the antimicrobial agents. In addition they can act as detergents, foaming agents and dispersants. The surfactants may be ionic surfactants (cationic surfactants, such as amine or quaternary ammonium surfactants, or anionic surfactants, such as sulfate, sulfonate, phosphate ester or carboxylate surfactants), non-ionic surfactants (such as fatty acid surfactants), or zwitterionic

(amphoteric) surfactants (such as phospholipids, phosphatidylserines,

phosphatidylethanolamines, phosphatidylcholines, and sphingomyelins). In some ebodiments, the surfactants are non-ionic surfactants. Preferred surfactants include alkyl polyglucosides, and fatty acid ethoxylates such as sorbitan monooleate and sorbitan trioleate. The surfactants may be present in an amount of about l-25wt%, preferably, about 7-20wt%, more preferably about 10-15wt% of the total composition.

Humectants are usually hygroscopic chemicals which help to keep the skin moist and increase the skin penetration of other ingredients. Suitable humectants include sugar polyols and sugar alcohols such as sorbitol, malitol, and glycerol. Humectants may be present in an amount of about 0.5-20wt%, more preferably about 1- 10 wt% most preferably about 3- 8wt% of the total composition.

Stabilisers maintain the function and activity of other ingredients such as, for example, antimicrobial agents, fragrances and essential oils. Suitable stabilisers include coconut oil derivatives. Stabilisers may be present in an amount of about 0.5-20wt%, more preferably about 1- 10 wt% most preferably about 3-8wt% of the total composition.

Preservatives are used to prevent contamination by the growth of microorganisms, and/or undesirable chemical changes. They help to maintain the contents of the composition over the shelf life of the product. Suitable preservatives are known to the skilled person. One suitable preservative is Acticide MBS. Preservatives are used in an amount necessary to provide the desired effect i.e. to maintain the composition during storage over a number of years e.g. 1-5 years. Preservatives may be present in an amount of about 0.01-20wt%, about 0.1- 10 wt%, about 0.5-8wt% or about 1.0-5.0 wt% of the total composition. The composition may also include other ingredients which can be beneficial to the health of users of the composition, or the surface or cleaning article sanitized by the composition.

These ingredients may help to maintain healthy skin by for example promoting cell proliferation and wound healing, increase skin smoothness, increase the moisture contact of the extracellular matrix. Preferably these ingredients are natural. Examples of suitable ingredients include allantoin and aloe vera extract.

Allantoin is present in botanical extracts of the comfrey plant and urine from most

mammals, notably cows. The most commonly used species of comfrey to obtain allantoin is Russian comfrey Symphytum xuplandicum which is a cross or hybrid of Symphytum officinale (Common Comfrey) and Symphytum asperum (Rough Comfrey). Allantoin is used as an active ingredient in over-the-counter cosmetics. The claimed beneficial effects of allantoin include increased moisture content of the extracellular matrix, enhanced desquamation of upper layers of dead skin cells, increased skin smoothness, promotion of cell proliferation and wound healing, soothing, anti-irritant and skin protectant effect by forming complexes with irritant and sensitizing agents

Aloe vera, called kathalai in Ayurvedic medicine, is used as a multipurpose skin treatment. This may be partly due to the presence of saponin, a chemical compound that acts as an antimicrobial agent. The aloe vera extract is derived from the inner leaves of the aloe Vera plant. Aloe vera water can be prepared by dissolving lOg spray dried aloe vera extract in about 2L deionised water.

As discussed above the antimicrobial agents may have other beneficial properties.

Additional ingredients can be selected to prepare compositions which provide specific desired effects. For example compositions to provide a calming effect may comprise

Fragonia, whilst compositions comprising tea tree oil and optionally rose geranium may promote skin healing. One preferred composition comprises Fragonia, aloe vera extract and allantoin. Another preferred composition comprises Tea Tree oil, allantoin, aloe vera extract and optionally Rose Germanium oil.

In one embodiment the composition comprises 2-75wt% alcohol, 0.5-5wt% antimicrobial agent and 24-97wt% aqueous media. The composition may comprise:

2-75wt% alcohol

l-5wt% antimicrobial agent 7-20wt% surfactant

l-10wt% emollient

l-10wt% stabiliser

0-0.05wt% essential oils

balance aqueous media

The antimicrobial agent may comprise Tea Tree oil and Fragonia oil, wherein the final composition comprises at least 0.5wt% of each component. In a preferred embodiment the composition comprises:

3wt% ethanol

0.1wt% allantoin

2wt% Glycerol

8wt% Polyoxyethylene (20) sorbitan monooleate

2wt% Tea tree oil

2wt% Fragonia oil

4.5 wt% polyglycoside surfactant based on a C8-C16 natural fatty alcohol 3 wt% Foam booster/stabiliser

balance aloe vera extract in water

In one embodiment, the composition may comprise about 55-70wt% alcohol and about 0.5-5wt% antimicrobial agent, with the balance made up to 100% with water or other aqueous media. The antimicrobial agent may comprise Tea Tree oil and Fragonia oil, wherein the final composition comprises at least 0.25wt% of both. In some embodiments of the invention, the Tea Tree oil and Fragonia oil are present in equal quantities.

In one embodiment, the composition may comprise 55-70wt% alcohol, 30-40 wt% aqueous media, and 0.5-5wt% antimicrobial agent. Preferably the composition comprises:

66.3wt% alcohol

0.6 wt% antimicrobial agent

0-0.05 wt% essential oils

balance aqueous media

All percentages of components for the compositions are wt% i.e. percentage by weight of the total composition. In embodiments of the invention, the composition will generally comprise an alcohol and the antimicrobial agent (optionally additional components, such as surfactants, essentials oils, emollient and/or stabilisers, as described above), with the balance of the composition made up of aqueous media, such as aloe vera extract in water, to complete the composition up to 100wt%. The water may be deionised water. In some embodiments of the invention, the antimicrobial agent in the composition comprises Tea Tree oil and Fragonia oil, for example at least 0.25wt%, at least 0.5wt% or at least lwt% of each of Tea Tree oil and Fragonia oil by weight of the final composition.

The composition of the invention is preferably a liquid, preferably a sprayable liquid. The composition may also be in the form of a gel or foam. The composition may form a spray or foam when dispensed for use. The composition may be dispensed from a pressurised container, such an aerosol can (and hence the container may further comprise a propellant), or from a container using a hand operated pump.

The compositions of the invention can be used to clean and/or sanitise body cleaning articles by bringing the composition into contact with the cleaning article. The composition may be sprayed onto the article and allowed to dry. Optionally the cleaning article may be squeezed or rubbed to increase penetration of the composition into the article. Alternatively the cleaning article can be immersed, wholly or partially, or soaked in the composition of the invention. The cleaning article is immersed for length of time, for example from 2-60 minutes, preferably 5-30 minutes, more preferably 10-15 minutes. The cleaning article can then be removed, optionally squeezed to remove excess composition and allowed to air dry.

The composition can be used after each use of the cleaning article. Alternatively the composition can be used daily, twice weekly, weekly, fortnightly, monthly or at any other time period.

The invention also provides the use of the composition as a cleaning and/or sanitising composition. Preferably the composition can be used to clean or sanitise a body cleaning article. Preferred features of the composition are as described above.

The invention also provides a method of cleaning or sanitising a body cleaning article, comprising contacting said body cleaning article with a composition, said composition comprising an antimicrobial agent dissolved in a mixture of an alcohol and an aqueous media. Preferred features of the composition are as described above. The step of contacting the article with the composition may comprise spraying the composition onto the article, or soaking and/or immersing the article in the composition. The article may be completely or partially immersed in the composition. In some embodiments, the article is completely or partially immersed in a dilution of the composition, for example ap to a 1:1, 1:2, 1:3, 1:4, 1:5, 1:10 or 1:20 composition:water dilution.

In another aspect of the invention there is provided a container, such as a bottle or spray can, comprising a cleaning composition of the invention. The container may be any suitable size, for example a 100ml, 250ml, 500ml or 1000ml container. The container may therefore comprise 100ml, 250ml, 500ml or 1000ml of the cleaning composition of the invention.

In a further aspect the invention provides a kit comprising (i) a container comprising a cleaning composition as described herein and (ii) a body cleaning article. Preferred features of the composition are as described above. The kit may further include instructions for use of the composition described herein.

Compositions can be used to sanitise surfaces. Suitable surfaces are preferably non-porous surfaces in domestic or industrial settings. Examples of suitable surfaces include food preparation areas such as kitchen work surfaces and hobs; tiles; floors; and sanitary ware, such a basins, showers, and toilets. Preferably the composition is an alcohol-based composition. The alcohol within the composition evaporates to leave a sanitising layer on the cleaned surface. The composition can be sprayed onto the surface and then spread or wiped over the surface using a conventional cleaning cloth or tissue. Alternatively the composition may be supplied in a fabric impregnated or soaked in the composition, for example in the form of a wipe.

The invention also provides the use of a composition as described herein for cleaning and/or sanitising a surface. Preferred features of the composition are as described above. Preferably the composition is an alcohol-based composition.

The invention also provides a method of cleaning a hard surface comprising contacting said hard surface with a composition of the invention. Preferred features of the composition are as described above. Preferably the composition is an alcohol-based composition. The composition may be sprayed onto the surface and then distributed using a fabric cloth or tissue such as those conventionally used in domestic cleaning. Alternatively the composition can be applied to a conventional cleaning cloth or tissue, which is then wiped over the surface. The surface may also be sanitised using a fabric cloth, such as a wet wipe, which is supplied impregnated with a composition of the invention.

Also provided is a wipe, such as a fabric wipe, impregnated with the composition of the invention. Preferred features of the composition are as described above. Preferably the composition is an alcohol-based composition. Such wipes are generally known as "wet-wipes". Materials and methods for producing such wipes are known in the art. Suitable materials for the wipe substrate include silk, keratin, cellulose (such as cotton), acetates, acrylics, polyesters, polyvinyl alcohols, nylon, polypropylene, polyethylene, polyurethane, polyvinyl chloride, rayon and mixtures thereof. The wipes may be woven or non-woven. Depending on the material used, typically wet wipes can be produced as air-laid paper where the fibres are carried and formed to the structure of paper by air. Alternatively wet wipes can be formed from synthetic fibres, or natural fibres such as cotton. The fibres may be woven together. The wipes are moistened or impregnated with the composition of the invention, preferably an alcohol-based composition. The wipe may be treated with softeners or lotions as known by the skilled person. The finished wet wipes can be folded and packaged singly or in multiple packs. The wipes can be packaged in flexible plastic packs or in a box dispenser, as is conventional for wet wipes.

The composition of the invention can be made by mixing all of the components together in a suitable vessel. The ingredients can be added sequentially followed by thorough mixing before the next ingredient is added. Alternatively some components can be blended together, and then added to other ingredients, which also may be in the form a blend of two or more ingredients.

In one embodiment of the invention there is provided a composition for decontamination, comprising alcohol and an antimicrobial agent in an aqueous medium, wherein the the antimicrobial agent comprises Tea Tree and Fragonia oil. The alcohol can be present in an amount of 55-70wt%. The antimicrobial agent can be present in an amount of about 0.5-5wt%, although the composition comprises at least 0.25% of each of the Tea Tree oil and Fragonia oil. Optionally, the Tea Tree oil and Fragonia oil are present in equal quantities. The balance of the composition to 100% is made up with the aqueous medium. Example compositions of the invention Ethanol-based sanitisers

Water-based compositions

The composition listed above was prepared as follows:

Part A

Aloe Vera water is prepared by dissolving 5g Aloe Vera powder in 1L deionised water at least three hours before using the solution to allow the Aloe Vera to fully hydrate. Initially the powder is dissolved in approximately half the required quantity of water. Preservative is added and the solution made up to volume with deionised water.

Part B

Part B is made in a separate vessel.

Allantoin and ethanol are mixed thoroughly until no powder remains.

Add glycerol and blend thoroughly.

Add Polyoxyethylene (20) sorbitan monooleate (Ecoteric T20) and blend thoroughly.

Add Fragonia and/or Tea Tree Oil and blend thoroughly.

Add any essential oil used for fragrance and blend thoroughly.

Part C

Add Aloe Vera water slowly to Part B. Initially, add dropwise stirring thoroughly.

When the volume of Aloe Vera water added is approximately equal to the initial volume of Part B, add polyglycoside surfactant based on a C8-C16 natural fatty alcohol (Glucopon 425 N) and stir thoroughly.

Add Foam stabiliser/booster (Hi Foam 750) and stir thoroughly.

Add Aloe Vera water until final volume is attained.

Add dye, if required, when volume is approximately 95wt% of the final volume.

Examples Minimum Inhibitory Concentration (MIC) of Tea Tree Oil, Fragonia Oil and Mixtures of Tea Tree Oil and Fragonia Oil against selected microorganisms

Overview

Tea Tree oil and Fragonia oil samples were tested as received to determine MIC against selected strains of bacteria and fungi. Mixed samples of Tea Tree oil and Fragonia oil (50/50 by weight) were tested to determine if there was any synergistic effect on M IC.

Samples

Approximately 5mL Tea Tree Oil and 5mL Fragonia oil were provided for testing. Preparation of Stock Solutions for Testing

Accurately weigh 2,000 ±10mg test sample (Fragonia oil or Tea Tree Oil) and mix thoroughly with 5g polysorbate 20. Diluted to lOOmL with deionised water. The stock solution contains 20,000ppm essential oil.

Preparation of Stock Solution of Mixed Essential Oils

Accurately pipette 20mL of the stock solution of Tea Tree oil and 20mL of Fragonia oil. Mix thoroughly. This solution contains l,000ppm of tea Tree oil and l,000ppm of Fragonia oil.

Preparation of Placebo Solution

A placebo (control) was prepared as described above omitting the essential oil.

Working Inoculum Preparation

Inoculate 2-3 bacterial colonies from an 18 hour plate culture into lOmL of Muller Hinton Broth (MHB). Vortex and incubate at 37±2°C for 6 hours. Adjust this to a 0.5 McFarland Standard with MHB. Check by measuring absorbance at 625nm and adjust to a final absorbance of 0.080-0.100. This is the working inoculum. Final bacterial suspension for testing requires a 1:500 dilution in MHB. Add O.lmL of working inoculum to 9.9mL of M HB. Then take 4 mL of this dilution and add to 16 mL of M HB to make the final inoculum (approx. 2 ^χ 10⁵ CFU/mL). Final bacterial suspension should be used within 15 minutes of preparation. Determine the total viable count (TVC) of the final bacterial suspension.

96 Well Plate Preparation:

Perform this step in a biological safety cabinet.

Note this test is adapted from standard test procedures. In practice only six dilutions were carried out per sample i.e. samples were only tested at 20,000, 10,000, 5,000, 2,500, 1,250 and 625ppm.

1. Add 10 μί of stock solution to Column 12 (20,000 ppm). Add 10 μί of serially diluted solutions to Columns 11 to 3 respectively. Columns 1 and 2 are negative and positive control wells - add 10 μί of placebo to each.

2. Pour Muller Hinton Broth into a sterile trough

3. Adjust the pipette to 90μί and dispense M HB from the trough into wells columns 1-12. (1 and 2 are positive and negative control wells)

4. Add ΙΟΟμί of final bacterial suspension to columns 2-12 (column 1 is the negative control). 5. Seal the tray carefully with plastic adhesive lid. Label appropriately. Make sure bacteria/test solutions are mixed adequately by tapping 96 well plates sharply a couple of times.

6. Incubate at 37±2°C for 24 hours. Stack no more than 4 plates high. Reading the Plate:

Read the plate at 450nm using an ELISA reader.

Determine any dramatic changes in absorbance. The MIC is the concentration of the first well that has the same absorbance as the negative control well.

Results

Table 1 Activity against Microorganisms

Table 2 Activity against Fungi

Discussion of Results

Note that the method of dilution meant that samples were tested at 20,000, 10,000, 5,000, 2,500, 1,250 and 625ppm. Finer gradations of activity are not be differentiated by this method. Further work is required in the range 2,500 to >5,000ppm to determine MIC values with greater precision.

The results against E coli and S Aureus were similar for Tea Tree and Fragonia oil. The results for the mixed solution clearly indicated a synergistic effect with a mixture of essential oils.

The results against the fungus A. niger suggested that Fragonia oil has higher activity against this organism than Tea Tree oil. Results for C. albicans were similar for both essential oils. There was clearly a synergistic effect in the case of C. albicans. Antiviral Activity of Tea Tree Oil, Fragonia Oil and Mixtures of Tea Tree Oil and Fragonia Oil against human Herpes simplex virus

Overview A representative surface is inoculated with the test viruses (Herpes simplex). The surface is then treated with a solution containing a known concentration of essential oil. The test solution is allowed to remain in contact with the test surface for approximately 15 minutes. Following treatment, the surface is assayed for surviving viruses by an assay method specific for the test virus.

Preparation of Test Solutions

Test solutions were prepared as described for M IC against microorganisms.

Preparation of Test Surfaces

The test surface is inoculated with a solution containing a known concentration of Herpes simplex virus.

Test Method

The test surface is sprayed with the test solution until saturated. Excess liquid is allowed to drain and the surface left undisturbed for 1 hour.

Assay

The number of viable particles was determined using a culture test method specific to the virus.

Results

Table 3 Activity against Herpes Simplex

Conclusion

The results indicate that Tea Tree oil and Fragonia oil are similar with respect to antiviral activity. The results also suggest that there is a synergistic effect when the two essential oils are mixed.

Claims

1. A cleaning composition comprising an antimicrobial agent dissolved in a mixture of an alcohol and an aqueous media, wherein said antimicrobial agent comprises Tea Tree oil and Fragonia oil.

2. A cleaning composition of claim 1 wherein said composition comprises:

2-75 %wt alcohol;

0.5-5 %wt antimicrobial agent; and

24-97 %wt aqueous media.

3. A cleaning composition of claim 1 or claim 2 wherein said composition comprises 2-75%wt alcohol;

l-5%wt antimicrobial agent;

7-20%wt surfactant;

l-10 wt emollient;

l-10 wt stabiliser;

0-0.05%wt essential oils; and

balance percentage aqueous media.

4. A cleaning composition as claimed in any preceeding claim comprising

3%wt ethanol;

0.1%wt allantoin;

2%wt Glycerol;

8%wt Polyoxyethylene (20) sorbitan monoledate;

2%wt Tea tree oil;

2%wt Fragonia oil;

4.5%wt alkyl polyglycoside surfactant based on a natural C8-C16 fatty alcohol;

3%wt Foam stabiliser/booster; and

balance aloe vera extract in water.

5. A cleaning composition as claimed in claim 1 or claim 2 comprising

55-70%wt alcohol;

30-40 wt aqueous media; and

0.5-5%wt antimicrobial agent.

6. A cleaning composition as claimed in claim 1, 2, or 5 comprising

66.3%wt alcohol;

0.6 %wt antimicrobial agent;

0-0.05 wt essential oils; and balance aqueous media.

7. A cleaning composition as claimed in any preceding claim comprising at least 1250ppm Tea Tree oil and 1250ppm Fragonia oil.

8. A composition as claimed in any preceding claim wherein the antimicrobial agent is natural.

9. A composition as claimed in any preceding claim wherein the antimicrobial agent further comprises Rose geranium.

10. A composition as claimed in any preceding claim wherein said composition further comprising one or more excipients.

11. A composition as claimed in claim 10 wherein the excipient is selected from essential oils, colourants, perfumes, emollients, surfactants, stabilisers.

12. A composition as claimed in any preceding claim wherein said composition is a water- based composition.

13. A composition as claimed in any preceding claim wherein said composition is an alcohol- based composition.

14. A composition as claimed in any preceding claim wherein said alcohol is ethanol.

15. A method of cleaning a body cleaning article, comprising contacting said body cleaning article with a composition, said composition comprising an antimicrobial agent dissolved in a mixture of an alcohol and an aqueous media.

16. A method as claimed in claim 15 wherein said composition is a composition as claimed in any one of claims 1 to 14.

17. A method as claimed in claim 15 or claim 16, wherein contacting said body cleaning article comprises spraying the composition onto said body cleaning article.

18. A method as claimed in any one of claims 15 to 17, wherein contacting said body cleaning article comprises soaking or wholly or partially immersing the body cleaning article in the composition.

19. Use of a cleaning composition as claimed in any one of claims 1 to 14 in cleaning and/or sanitising a body cleaning article or a surface.

20. A kit comprising a body cleaning article and a container comprising a cleaning composition as claimed in any one of claims 1 to 14.