Title of Invention	METHOD TO ASSESS SURFACTANT ADSORPTION ON SKIN
Abstract	A convenient to use test method for comparing the relative affinity of surfactants for the skin employs a solid or semisolid matrix for associating a dye or pigment with the skin. The dye or pigment has an affinity to either the skin or a surfactant adsorbed on the skin during personal cleansing. The matrix may include a reagent soluble polymer film or temporary tattoo.

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J9001/C FORM - 2 THE PATENTS ACT, 1970 (39 of 1970) & The Patents Rules, 2003 COMPLETE SPECIFICATION (See Section 10 and Rule 13) METHOD TO ASSESS SURFACTANT ADSORPTION ON SKIN HINDUSTAN UNILEVER LIMITED, a company incorporated under the Indian Companies Act, 1913 and having its registered office at Hindustan Lever House, 165/166, Backbay Reclamation, Mumbai -400 020, Maharashtra, India The following specification particularly describes the invention and the manner in which it is to be performed WO 2007/104396 PCT/EP2007/001165 - 1 - METHOD TO ASSESS SURFACTANT ADSORPTION ON SKIN BACKGROUND OF THE INVENTION 5 Field of the Invention The invention relates to a" test method for estimating the level of residual surfactant on the skin. 10 The Related Art Personal cleansing products are frequently marketed based on their degree of mildness to the skinof the user. Unfortunately, many surfactants in common use tend bind to 15 the skin proteins, which can result in skin irritation. Moreover different surfactants bind to skin proteins to varying degrees. Cleansers that do not leave residual surfactants on the skin are therefore more desirable to many consumers. At least one test has been disclosed for 20 evaluating the amount of dye and/or pigment binding to skin and evaluating the extent of skin adsorption of surfactant which interferes with dye and/or pigment binding. However there has not been a convenient way for the consumer to ascertain how a particular cleanser will interact with their 25 skin using a particular cleansing method until now. A diagnostic test is disclosed by Imokawa and Mishima, Contact Dermatitis 5:357-366, (1979) which uses dye to determine whether or not skin has appreciable amounts of 30 adsorbed surfactant. The disclosed test uses dye solutions WO 2007/104396 PCT/EP2007/001165 - 2 - which are not convenient or appropriate when developing a convenient method intended for consumer use. A convenient method for detecting and quantifying residual 5 surfactants has been discovered that employs soluble or dispersible solid or semisolid films as dye and/or pigment carriers where the dye and/or pigment(s) have an affinity for either the skin or the surfactant left on the skin. Although not wishing to be bound to the following theory, it 10 is believed that in a first case where a dye and/or pigment has an affinity for the skin but not for the residual surfactant of interest, application of the dye and/or pigment to skin with residual surfactant will result in skin with less coloration then to skin without the residual 15 surfactant. In a second case where a dye and/or pigment has an affinity for the residual surfactant of interest but not for the skin, application of the dye and/or pigment to skin with residual surfactant will result in skin with more coloration then to skin without the residual surfactant. 20 Both cases assume that neither the surfactant nor the reagent which removes the film/coating alters the skin itself in a way that changes the skin's propensity to interact with a given dye or pigment independently of the amount of surfactant adsorbed onto the skin. 25 SUMMARY OF THE INVENTION In one aspect of the invention is a method for estimating the residual amount of a cleansing composition remaining on 30 skin of a user after washing, including but not limited to the steps of: WO 2007/104396 PCT/EP2007/001165 - 3 - a. treating an area of skin by applying the cleansing composition thereto wherein the cleansing composition contains at least about 1 % by wt. of an anionic surfactant; 5 b. removing said cleansing composition from the treated area of the skin until there is no longer perception of any residual cleansing composition remaining on the treated area of the user's skin; c. applying to the treated area of the skin, a solid 10 or semisolid film or coating that is soluble or dispersible in a first reagent, the film or coating containing at least 0.001 % by wt. of dye and/or pigment(s), wherein said dye and/or pigment(s) are capable of adsorbing, reacting and/or associating with either the anionic surfactant or human or is animal skin d. contacting the film or coating with sufficient first reagent effective to allow the dye and/or pigment contained therein to contact the treated area of the skin; and 20 e. assessing the amount of dye and/or pigment adsorbed onto either the skin or residual surfactant upon the skin upon removal of the film or coating with the first reagent. 25 In another aspect of the invention is a diagnostic kit for estimating the residual amount of a cleansing composition containing one or more anionic surfactants remaining on the skin after removal of the cleansing composition, the kit including but not limited to the following: 30 WO 2007/104396 PCT/EP2007/001165 - 4 - a. a solid or semisolid film that is soluble or dispersible in a first reagent containing at least 0.001 % by wt. of dye and/or pigment(s) , wherein said dye and/or pigment(s) is/are capable of adsorbing, reacting and/or 5 associating with either the anionic surfactant or human or animal skin;, b. instructions for applying the film to the skin and visualizing the residual adsorbed anionic surfactant on the skin after removal of the film; and 10 c. a color gradation scale denoting the intensity of adsorbed dye and/or pigment on the skin corresponding with the approx. levels of residual cleansing composition on the skin. 15 Preferred embodiments of the invention will now be described by way of example with reference to the accompanying figures. BRIEF DESCRIPTION OF THE FIGURES 20 Fig. 1 is a bar graph depicting the color measurement (L*) of surfactant type binding using indigo carmine staining intensity of treated skin tested according to the procedure of Example 1. 25 Fig. 2 is a photograph of a subject's forearm tested with various surfactant solutions according to the procedure of Example 1 that is depicted in Fig. 1. 30 Fig. 3 is a bar graph depicting the color measurement (L*) of surfactant binding using commercially available toilet WO 2007/104396 PCT/EP2007/001165 - 5 - bars and indigo carmine staining intensity of treated skin tested according to the procedure of Example 1. Fig. 4 is a photograph of a subject's forearm tested with 5 various commercially available toilet bars according to the procedure of Example 1 that is depicted in Fig. 3. DETAILED DESCRIPTION OF THE INVENTION 10 In one aspect of the invention is a method for estimating the residual amount of a cleansing composition remaining on skin of a user after washing, including but not limited to the steps of: a. treating an area of skin by applying the cleansing 15 composition thereto wherein the cleansing composition contains at least about 1, 2, 3, 4 or 5 % by wt. of an anionic surfactant; b. removing said cleansing composition from the treated area of the skin (preferably by rinsing with water) 20 until there is no longer perception of any residual cleansing composition remaining on the treated area of the user's skin (advantageously followed by pat drying the treated area); c. applying to the treated area of the skin a solid 25 or semisolid film or coating that is soluble or dispersible in a first reagent, the film or coating containing at least 0.001 % by wt. of dye and/or pigment(s) wherein said dye and/or pigment(s) is/are capable of adsorbing, reacting and/or associating with either the anionic surfactant or 30 human or animal skin,-, (e.g. indigo carmine, octadecyl WO 2007/104396 PCT/EP2007/001165 - 6 - fluorescein, beta carotene, lycopene and/or methylene blue and the like) d. contacting the film or coating with sufficient first reagent effective to allow the dye and/or pigment 5 contained therein to contact the treated area of the skin; and e. assessing the amount of dye and/or pigment adsorbed onto either the skin or residual surfactant upon the skin upon removal of the film or coating. io In one preferred embodiment, the dye and/or pigment adsorbed onto either the skin or residual surfactant upon the skin is further treated with sufficient second reagent having an affinity for the dye and/or pigment to remove excess dye 15 and/or pigment. Advantageously the amount of dye and/or pigment adsorbed onto the treated area of the skin is visually compared to a control treated area of skin containing no adsorbed surfactant. Preferably the amount of dye and/or pigment adsorbed onto the treated area of the 20 skin is compared to a control treated area containing no adsorbed surfactant by a quantitative energy reflectance and/or absorption method(s). More preferably the film is selected from a self-supporting film (such as a water, alcohol or oil soluble polymeric film) or a non-self- 25 supporting coating (such as a temporary tattoo). In a preferred embodiment, the cleansing composition contains at least 5 % of anionic surfactant(s) having an individual zein value of greater than about 10, 20, 30, 40 30 or 50. Advantageously the cleansing composition contains one or more anionic surfactants selected from acyl WO 2007/104396 PCT/EP2007/001165 - 7 - glutamates, acyl peptides, sarcosinates, taurates, alkanoic acids and alkanoates, ester carboxylic acids, ether carboxylic acids, phosphoric acid esters and salts, acyl isethionates, alkylaryl sulfonates, alkyl sulfonates, 5 sulfosuccinates, alkyl ether sulfates, or alkyl sulfates or a blend thereof. Preferably the dye and/or pigment is present in the film in the concentration range of about 0.001 to 50 wt. % 10 preferably at a lower limit of about 0,01, 0.05 or 0.1 to an upper limit of about 0.5, 1, 2, 3, 4, 5, 10, 20 or 30 % by wt. More preferably the dye and/or pigment(s) is/are selected from one or more of the following classes: acid, basic, mordant, natural, solvent, direct, FD&C dyes, D & C 15 dyes. Suitable pigments include those generally recognized as safe, and listed in C.T.F.A. Cosmetic Ingredient Handbook, First Edition, Washington D.C. (1988), incorporated herein by reference. 20 Advantageously, the film contains one or more of the following components: carbohydrate(s) , protein (s), acrylamide(s) , or quaternary modified compounds or blends thereof (advantageously starch, cellulose, hydroxypropyl trimonium guar, pullulan, gellan gum, polyguaternium-22, 25 gelatin, polyacrylamide and the like). Preferably the reagent contains one or more of the following components: water, mono and/or polyhydric alcohol(s); polyol{s); mineral, glyceride and/or silicone oils, or a blend thereof. 30 In another aspect of the invention is a diagnostic kit for estimating the residual amount of a cleansing composition WO 2007/104396 PCT/EP/2007/001165 - 8 - containing one or more anionic surfactants remaining on the skin after removal of the composition, the kit including but not limited to the following: 5 a. a solid or semisolid film that is soluble or dispersible in a first reagent, the film containing at least 0.001 % by wt. of a dye and/or pigment(s), wherein said dye and/or pigment(s) is/are capable of adsorbing, reacting and/or associating with either the anionic surfactant or io human or animal skin;, {e.g. indigo carmine, octadecyl fluorescein, beta carotene, lycopene and/or methylene blue and the like) ,- b. instructions for applying the film to the skin and visualizing the residual adsorbed anionic surfactant on 15 the skin after removal of the film and optionally of excess dye and/or pigment(s); and c. a color gradation scale denoting the intensity of adsorbed dye and/or pigment on the skin corresponding with the approx. levels of residual cleansing composition on the 20 skin. Dyes and pigments Suitable dyes and/or pigments include various organic and 25 inorganic dyes or pigments or blends thereof that have an affinity for and/or color the residual cleansing composition or the skin itself. Suitable organic pigments include azo, indigoid, triphenylmethane, anthraquinone, and xanthine dyes which are designated as D&C and FD&C (blues, browns, greens, 30 oranges, reds, yellows, etc.). Inorganic pigments are generally insoluble metallic salts of certified color WO 2007/104396 PCIYEP2007/001165 - 9 - additives, referred to as lakes or iron oxides. Other suitable pigments are selected from the group of titanium dioxide, zinc oxide, and metallic lakes of cosmetic dyes or blends thereof. Suitable natural dyes include turmeric 5 oleoresins, cochineal extracts, gardenia extracts, and natural colors derived from vegetable juices. Other specific examples of suitable natural dyes include, but are not limited to, beet extract, grape skin extract, and chlorophyll containing extracts (e.g. nettle extract, 10 alfalfa extract and spinach extract). Suitable pigments include those generally recognized as safe, and listed in C.T.F.A. Cosmetic Ingredient Handbook, First Edition, Washington D.C. (1988), incorporated herein by reference. Specific examples are red iron oxide, yellow iron oxide, 15 black iron oxide, brown iron oxide, ultramarine, FD&C Red, Nos. 2, 5, 6, 7, 10, 11, 12, 13, 30 and 34; FD&C Yellow No. 5, Red 3, 21, 27, 28, and 33 Aluminum Lakes, Yellow 5, 6, and 10 Aluminum Lakes, Orange 5 Aluminum Lake, Blue 1 Aluminum Lake, Red 6 Barium Lake, Red 7 Calcium Lake, and 20 the like. Reagent soluble or dispersible solid or semisolid films: In a preferred embodiment, a solid or semisolid film or 25 coating that is soluble or dispersible in a first reagent is a useful means of delivering the dye(s) and/or pigment (s) to the skin as described above. A solid or semisolid film or coating that is soluble or dispersible in a first reagent is herein defined as that which dissolves or disperses over a 30 period of time when contacted by the first reagent at 25 C. Upon film dissolution, the dye(s) and/or pigment(s) WO 2007/104396 PCTYEP2007/001165 - 10 - contained in the film or coating contact the skin area on which the film or coating has been placed and are visible. Advantageously this time can be in the range of about 0.5 to 10 seconds while contacted with the first reagent. The time 5 may be shortened if the first reagent is used at an elevated temperature. Release is here defined as allowing at least about 50 % of the dye(s) and/or pigment{s) in the film to contact the skin next to the film in about 5 seconds when the film is contacted by a suitable first reagent having an 10 affinity for the film at 25 C. When the first reagent contains water, materials that may be used to make the water soluble or dispersible solid or semisolid films or coatings of this invention include well 15 known water soluble or dispersible resins, such as pullulan, other water soluble/dispersible polysaccharides and derivatives thereof, water soluble/dispersible cellulosic derivatives and the like. Other suitable resins are described in Davidson and Sittig, Water - Soluble Resins, 20 Van Nostrand Reinhold Company, New York (1968), herein incorporated by reference. The water-soluble or dispersible resin should have useful characteristics such as sufficient water solubility or dispersibility so as to quickly release the dye and/or pigment contained within the film, and 25 sufficient strength and pliability in order to permit machine handling while trimming and packaging without tearing. Other preferred water-soluble or dispersible resins include polyvinyl alcohol, cellulose ethers, polyethylene oxide, starch, polyvinylpyrrolidone, 30 polyacrylamide, polyvinyl methyl ether-maleic anhydride, polymaleic anhydride, styrene maleic anhydride. WO 2007/104396 PCT7EP2007/001165 - 11 - hydroxyethylcellulose, methylcellulose, polyethylene glycols, carboxymethylcelulose, polyacrylic acid salts, alginates, acrylarnide copolymers, guar gum, casein, ethylene-maleic anhydride resin series, polyethyleneimine, 5 ethyl hydroxyethylcellulose, ethyl methylcellulose, hydroxyethyl methylcellulose and the like. Other useful films include those that are either oil soluble/dispersible, alcohol soluble/dispersible, or 10 soluble/dispersible in any reagent(s) that is/are safe for contact with human or animal skin. Oils may consist of one or more mineral, glyceride, silicone or any other suitable oil or a blend thereof. Alcohols may include one or more monohydric or polyhyric alcohols, polyols or blends thereof 15 and the like. Suitable film examples include water/alcohol-soluble or water/alcohol-dispersible polymers that can be fashioned into films such as octylacrylamide/acrylates/butylaminoethyl methacrylate copolymer neutralized with a base such as 20 aminomethylpropanol, alcohol-soluble polyamides, polyethers of 2,2- bis (4-hydroxyphenyl) propane and epichlorohydrin, and the like. Suitable oil soluble polymers that can be used in films include vinyl polymers such as polymers derived from the polymerization of at least one monoethylenically 25 unsaturated alkyl (meth) acrylic monomer, preferably, an alkyl (meth)acrylic acid ester (i.e., an alkyl acrylate or alkyl methacrylate) , wherein the alkyl group has at least 4 carbon atoms (on average) and preferably no greater than 22 carbon atoms (on average), and at least one 30 monoethylenically unsaturated poly(alkylene oxide) monomer, preferably, a monoethylenically unsaturated poly(alkylene WO 2007/104396 PCT/EP2007/001165 - 12 - oxide) (meth)acrylic monomer and the like. Depending on the properties of the resultant polymer, the monoethylenically unsaturated alkyl (meth)acrylic acid esters used to prepare the polymer can have just short alkyl groups {e.g., at least 5 4 carbon atoms (on average) and preferably no greater than 14 carbon atoms {on average)), or just long alkyl groups (e.g., at least 15 carbon atoms (on average) and preferably no greater than 22 carbon atoms (on average)), or mixtures of monoethylenically unsaturated alkyl (meth)acrylic acid 10 esters with short alkyl groups can be used in combination with monoethylenically unsaturated alkyl (meth)acrylic acid esters with long alkyl groups (e.g., terpolymers). The alkyl group can optionally contain heteroatoms (e.g., N, O, or S) and can be linear, branched, or cyclic. Other suitable 15 vinyl polymers include polyethylene having a molecular weight of 2000 to 8000, polystyrene or poly alpha methylstyrene, block copolymers of styrene with ethylene, propylene and/or butalene, copolymers of polyvinylpyrrolidone with polyethylene and the like. 20 Polymers such as cellulose and starch polymeric derivatives that are oil soluble, such a dextrin esters, e.g. dextrin palmitate or other oil-soluble polymers that may be fashioned into films may also be used. 25 Temporary tattoos In a second preferred embodiment, a temporary tattoo may be advantageously employed to ascertain the residual surfactant 30 level on the skin. The dye(s) and/or pigment(s) will advantageously have an affinity, for either the surfactant or WO 2007/104396 PCT/EP2007/001165 - 13 - the skin and will be incorporated in the temporary tattoo. There are three principle types of temporary tattoo which differ in the way they are applied to the skin. In a non-limiting manner, it is useful to classify temporary tattoos s as: 1) those which are hand-painted or drawn directly on the skin; 2) those which are printed (or hand- painted or drawn) onto a substrate and then transferred to the skin some time later; and 3) those which are printed (or hand-painted or drawn) onto a transparent substrate which is 10 subsequently adhered onto the skin. Preferably the transparent substrate is soluble or dispersible in water, alcohol, oil or another reagent that is safe for skin contact. is For a type 1 temporary tattoo, the artwork is applied to the skin with brushes and/or pens or similar implements, using inks or dye(s) and/or pigments, in a fashion analagous to making an illustration on paper. A stencil can be used. For a type 2 temporary tattoo, the artwork is usually 20 printed in ink or dye and/or pigment onto a substrate (such as paper) having properties which allow the artwork to become dissociated from the substrate, some time after the printing, and transferred to skin {such examples have alternative names such decalcomania, decal or transfer). 25 This transference is most commonly achieved by placing the printed substrate on the skin with the substrate uppermost and then wetting the substrate and applying gentle pressure before peeling off the substrate, leaving the artwork adhered to the skin. Clearly, the substrate, inks and dye 3 0 and/or pigments must be selected such that they have properties which are appropriate to the transference WO 2007/104396 PCT/EP2007/001165 - 14 - procedure. Alternatively, there may be a transparent layer which covers the surface of the substrate and on top of which the artwork is printed. In this case, the transparent layer is transferred to the skin along with the artwork and 5 thereafter forms a protective cover for the artwork, being uppermost on the skin when the substrate is peeled away. Alternatively, a transparent protective layer may be applied to the tattoo after transfer of the artwork to the skin by covering the artwork with a strip of transparent material or 10 else by applying the transparent layer in the form of a solution from which the layer is deposited on the tattoo after evaporation of the solvent. A transparent protective layer may also be applied to a type 1 temporary tattoo by these methods. For a type 3 temporary tattoo, the artwork 15 is usually printed in ink or dye and/or pigment onto a transparent substrate which has an adhesive area that exceeds, overlaps or surrounds the artwork and assists, or is responsible for, its adherence to the skin (these may be alternatively called stickers). A type 3 temporary tattoo is 20 adhered onto the skin such that the artwork is in contact with the skin and the transparent substrate often provides a protective adhesive layer. A temporary tattoo (of any type) which is to be used for 25 skin testing according to the invention may also be produced so that certain designated areas of the artwork contain different test dye and/or pigment compounds. Different areas could also contain the same test dye and/or pigment compound or material but at different concentrations. The 3 0 person carrying out the test would in one embodiment keep a careful record or "tattoo map" detailing the precise WO 2007/104396 PCI7EP2007/001165 - 15 - distribution of the test dye and/or pigment materials in the tattoo. After the time allotted for the test, the tattoo may be partially or completely removed with an appropriate reagent or blend of reagents and the skin underneath 5 examined for any signs of reaction or adsorption of the dye and/or pigment. Comparing signs of residual surfactant or the lack thereof (e.g. skin coloration) with the map will enable the estimation (and optionally the distribution) of the residual surfactants on the skin. 10 Except in the operating and comparative examples, or where otherwise explicitly indicated, all numbers in this description indicating amounts of material ought to be understood as modified by the word "bout". 15 The following examples will more fully illustrate the embodiments of this invention. All parts, percentages and proportions referred to herein and in the appended claims are by weight unless otherwise illustrated. Physical test 20 methods are described below: Example: 1 Indigo carmine (2-(1,3-Dihydro-3-oxo-5-sulfo-2H-indol-2-25 ylidene)-2,3-dihydro-3-oxo-lH-indole-5-sulfonic acid disodium salt), an acid dye also known as FD&C Blue No. 2 (C.I. 73015), was employed in solution form to compare levels of residual surfactants remaining on the skin of three individual panelists after cleansing using both pure 30 surfactant sample solutions and also samples of various commercially available toilet bars on multiple sites of WO 2007/104396 PCT/EP2007/001165 - 16 - their forearms according the procedure described below and the results are summarized in Figs. 1 to 4 and Table 1. It was found that the dye coloration intensity increased with decreasing surfactant binding i.e. sodium dodecyl sulfate 5 (SDS) (least stain uptake) > Sulfosuccinate > Water > Alkyl polyglucoside (APG) (most stain uptake). While not wishing to be bound by the following theory, it is believed that lipid removal by APG directly from the skin results in exposure of more skin protein binding sites to the dye, 10 hence the greater intensity of coloration independent of surfactant adsorbtion. Another way of expressing the foregoing sequence {except for APG) is as follows: a. Little to no staining: Greatest surfactant binding; b. Weak staining: Moderate surfactant binding; c. Strong staining: 15 Less surfactant binding; d. Strong staining: Water Control. The actual skin staining results are illustrated in Fig. 2. Especially indicative of surfactant/dye binding is the b* value, the lower the number means the skin is bluer i.e. as 20 the b* scale decreases the color changes from yellow to blue so a lower number means bluer. It was expected that the water treated site should have the lowest number as this situation allows for the most dye molecules to bind to the skin (in the absence of adsorbed surfactant). The untreated 25 site, which had no stain applied, was expected to have the largest b* value (most yellow). Since soap molecules are known to bind to most of the skin proteins there is little opportunity for the dye to bind to the skin. The soap treated site will therefore be similar to the untreated 30 control site. Dove(r) was found to be slightly better than Olay(r) with respect to surfactant binding to the skin. Table 1. L*, a*, and b* colorimetric skin values of three panelists whose forearms were treated several locations with selected toilet bars vs. water and a control according to the forearm was procedure described below. Olay © Bar (5%) Ave. Dove ® Bar (5%) Ave. water treated Ave. untreated Ave. Soap (Zest®) Ave. Panelist L* A* 64 1.63 63.57 1.83 62.82 1.79 63.46 1.75 63.57 0.89 62.78 0.51 62.84 0.02 63.06 0.47 59.87 -1.94 59.97 -2.26 60.24 -2.25 60.03 -2.15 66.37 4.8 65.26 4.75 66.19 4.48 65.94 4.68 1 B* 13.55 13.43 13.49 13.49 11.68 10.57 10.59 10.95 7.46 7.72 7.82 7.67 14.7 14.48 14.59 14.59 Panelist L* A* 60.42 1.53 60.96 1.85 60.76 2.03 60.71 1.80 59.63 1,68 61.48 1.12 61.96 1.45 61.02 1.42 59.55 0.36 59.13 0.47 59.71 0.15 59.46 0.33 63.52 7.49 63.35 6.21 64.32 5.59 63.73 6.43 2 B* 16.35 13.75 13.86 14.65 13.17 12-77 13.89 13.28 11.58 11.62 11.48 11.56 16.43 15.84 15.47 15.91 Panelist L* A* 59.37 4.97 58.8 5.64 58.53 5.63 58.90 5.41 60 4.87 59.83 4.87 60.06 4.85 59.96 4.86 58.74 1.6 59 1.55 58.8 1.49 58.85 1.55 61,32 7.72 60.73 8.8 60.8 9.28 60.95 8.60 56.3 11.45 57.17 10.32 56.76 11.04 56.74 10.94 3 B* site 1 15.75 16.15 16.31 16.07 Site 2 14.4 14.56 14.56 14.51 Site 3 11.57 11.38 11.58 11.51 Site 4 17.21 18.12 18.9 Site 5 IB.08 16.99 16.7 16.9 16.86 WO 2007/104396 PCT/2007/001165 - 18 - Expressed in order of increasing intensity of dye staining, the results of the toilet bar testing are as follows: Ivory ® Little to no staining: (Greatest surfactant binding); Dove ® Moderate staining (Less surfactant binding); Olay ® 5 Moderate staining: (Less surfactant binding); and Water -Strongest staining: Water Control (no surfactant binding). While not wishing to be bound by the following theory, it is believed that soap binds to the skin more strongly than sodium cocoyl isethionate (SCI) found in syndet toilet bars o such Dove(r) and Olay(r) . Soap is also known to be more irritating than SCI. Note: Ivory® and Olay® toilet bars are manufactured by Procter & Gamble Inc. (Cincinnati, OH) and Dove® toilet bars 5 are manufactured by Unilever Inc. (Greenwich, CT.) Indigo Carmine forearm washing procedure Treat volar forearms (3.14 cm2 area) with 2 ml of o surfactant/product solution or slurry for 2 minutes, e.g. • 1% w/w surfactant (SDS, Sulfosuccinate, APG) 8% w/w bar slurry (Ivory (r) , Dove (r), Olay (r)) s • Remove surfactant and rinse site with 2 ml of DI water (at approx. 23 C), then repeat rinse step for bars. Pat area dry • Apply 2 ml of 1% indigo carmine dye (aqueous) for 1 minute WO 2007/104396 PCT/EP2007/001165 - 19 - • Remove dye and rinse site with 2 ml of DI water (at approx. 23 C) Pat area dry • Obtain digital images with a conventional digital 5 camera. • Measure l*a*b* values at test sites using CR-10 chromameter (Minolta CM 508D spectrophotometer) Example 2: ■ 10 Samples of a commercially available water dissolvable film (1) comprised of Pullulan and containing a food grade dye (FD&C Green No. 3) were applied to the skin after cleansing with several cleansers according to Table 2 , using the 15 procedure described below. The results of the study are provided in table 1. Note (1) Listerine PocketPaks ® Oral Care Strips (available from Warner-Lambert Company Inc. Table 2 Tested Avg *b Dove(r) (5%) 12.3 high stain Ivory(r) (5%) 17 low stain Untreate d 18.5 low stain Method: Listerine® strip application WO 2007/104396 PCT/EP2007/001I65 - 20 - Treat volar forearms (3.14 cm2 area) with 2 ml of surfactant/product solution or slurry for 2 minutes, e.g. • 1% w/w surfactant (SDS, Sulfosuccinate, APG). 8% w/w bar slurry (Ivory(r), Dove(r), Olay(r)). Remove surfactant and rinse site with 2 ml of DI water (at approx. 23 C) (repeat rinse step for bars). Pat area dry. 10 - After arms have been treated with specified products (see above procedure for example 1) take approximately 5 mi's of water (at approx. 23 C)and wet treated area. - Gently place a dry test strip over treated and wetted area 15 on forearm. - Gently pat test strip against skin with finger - Allow strip to remain in contact with forearm for 3 minutes, (or until arm has air dried). - After three minutes place area of forearm with test strip 20 in running stream of warm water (approx. 10 seconds at 40 - 50 C) and remove from the skin the remainder of the strip which has not dissolved. - Pat dry. WO 2007/104396 PCT/EP2007/001165 - 21 - Example 3: Table 3. Composition of a suitable type 1 temporary tattoo solution. Ingredient INCI Name Trade Name % Active (of Raw) % of Ingredi ent in Formula AS IS % Active {in final formula) Range Formula Preferred Most preferred Polyquatern ium-22 Merquat 280 41% 45% 18.4500% 0 to 4 0 % 5 to 30 % 15 to 20% Hydroxyethy lcellulose Natroso 1 250 HMR 1% 45% 0.4500% 0 to 2 % 0 to 1% 0.2 to 0.8% Indigo Carmine 0.01 to 10% 0.1 to 20 1 to 5% Water QS to 100% Procedure: A type 1 temporary tattoo having the composition given in table 3 may be painted on treated skin using a decorative 10 stencil (e.g. shaped like a butterfly). Prior to the application of the tattoo, the skin is treated with various toilet bars described in table 4. The site of the temporary tattoo is then exposed to warm (approx 50 - 60 C) running water for about 30 seconds and the skin's 1* a* and b* 15 properties are measured with a suitable spectrophotometer as described above. It is expected that the results listed in table 4 will be obtained. WO 2007/104396 PCT/EP2007/001165 - 22 -Table 4 Avg *b 01ay ® Bar (5%) ■ 12 low- mod value -mod-high stain Dove ® Bar (5%) 10 Low value -mod high stain water treated 8 Lowest value -most stain untreated 16 Highest Value -low stain Soap (e.g. Zest ®) 16 Highest Value -low stain 5 Zein test method The cleanser containing the anionic surfactant to be tested preferably have a zein solubility of greater than 10, 20, 30, 40 or 50 using the zein solubility method set forth 10 below. The greater the zein score, the more harsh the cleansing base/surfactant is considered to be. This method involves measuring the solubility of zein (corn protein) in cleansing base solutions containing the anionic surfactant of interest as follows: 15 0.3 g of cleansing base and 29.7 g of water are mixed thoroughly. To this is added 1.5 g of zein, and mixed for 1 hour. The mixture is then centrifuged for 30 minutes at 3000 rpm. After centrifugation,. the pellet is extracted, 20 washed with water, and dried in a vacuum oven for 24 hours until substantially all the water has evaporated. The weight of the dried pellet is measured and percent zein solubilized is calculated using the following equation: . WO 2007/104396 PCT/EP2007/001165 - 23 - % Zein solubilized=100 (1-weight of dried pellet/1.5) . The % Zein is further described in the following references: E. Gotte, Skin compatibility of tensides measured by their 5 capacity for dissolving zein protein, Proc. IV International Congress of Surface Active Substanes, Brussels, 1964, pp 83-90. While this invention has been described with respect to io particular embodiments thereof, it is apparent that numerous other forms and modifications of the invention will be obvious to those skilled in the art. The appended claims and this invention generally should be construed to cover all such obvious forms and modifications which are within 15 the true spirit and scope of the present invention. - 24 - Claims A method for estimating the residual amount of a cleansing composition remaining on skin after washing, comprising the steps of: a. treating an area of skin by applying the cleansing composition thereto wherein the cleansing composition contains at least l% by wt. of an anionic surfactant; b. removing said cleansing composition from the treated area of the skin until there is no longer any perception of residual cleansing composition remaining on the treated area of the skin; c. applying to the treated area of the skin a solid or semisolid film or coating, that is soluble or dispersible in a first reagent, containing an effective amount of dye and/or pigment (s) wherein said dye and/or pigment(s) are capable of adsorbing, reacting and/or otherwise associating with either the anionic surfactant or human or animal skin and is/are present in a sufficient amount capable of detection in its associated state on the skin; d. contacting the film or coating with sufficient first reagent effective to allow the dye and/or pigment contained therein to contact the treated area of the skin; and e. assessing the amount of dye and/or pigment adsorbed onto either the skin or residual surfactant upon the skin upon removal of the film or coating. wherein the amount of dye and /or pigment absorbed onto the treated area of the skin is compared to a control treated area of skin containing no absorbed surfactant, either visually or by a quantitative energy reflectance and /or absorption method. AMENDED SHEET WO 2007/104396 PCT/EP2007/001165 - 25 - WO 2007/104396 PCT/EP2007/001165 - 26 - sulfonates, sulfosuccinates, alkyl ether sulfates, or alkyl sulfates or a blend thereof. 6, The method of claim 1 wherein the dye and/or pigment is present in tne film in the concentration range of 0.001 to 50 wt. %. 10 . 9. The method of claim 1 wherein the .dye and/or pigment (s) 7• is/are selected from one or more of the following classes: acid, basic, mordant, natural/ solvent, direct, FD&C dyes, D & C dyes. 10 . The method of claim 1 wherein the filmcontains one or 8 more of the following components: carbohydrate(s), 15 protein(s), acrylamide(s) , or quaternary modified compounds or blends thereof. 11 The method od claim 2 wherein the first reagent may be 9• the same or different compared to the second reagent and 20 wherein either the first or second reagent or both contain one or more of the following components: water, mono and/or polyhydric alcohol(s) ; poiyols, mineral, glyceride and/or silicone oils, or a blend thereof, 25 12. A diagnostic kit for estimeating the residual amount of a 10. cleansing composition containing one ormore anionic surfactants remaining on the skin after removal of the cleansing composition, the kit comprising: 30 a. a solid or semisolid film or coating, that is soluble or dispersible in a first reagent, containing at least 0.001 % by wt• of a dye and/or - 27 - pigment(s), wherein said dye and/or pigment(s) is/are capable of adsorbing, reacting and/or associating with either the anionic surfactant or human or animal skin; 5 b. instructions for applying the film to the skin and visualising the residual adsorbed anionic surfactant on the skin after removal of the film and optionally of excess dye and/or pigment(s); and c. a color gradation scale denoting the intensity of 10 adsorbed dye and/or pigment on the skin corresponding with the approx. levels of residual cleansing composition on the skin. Dated this 28th day of August 2008 HINDUSTAN UNILEVER LIMITED

Documents:

1858-MUMNP-2008-ANNEXURE TO FORM 3(10-4-2012).pdf

1858-MUMNP-2008-ANNEXURE TO FORM 3(14-1-2013).pdf

1858-MUMNP-2008-ASSIGNMENT(28-8-2008).pdf

1858-MUMNP-2008-CLAIMS(AMENDED)-(22-6-2012).pdf

1858-MUMNP-2008-CLAIMS(MARKED COPY)-(22-6-2012).pdf

1858-mumnp-2008-claims.doc

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1858-MUMNP-2008-CORRESPONDENCE(13-1-2011).pdf

1858-MUMNP-2008-CORRESPONDENCE(26-3-2013).pdf

1858-MUMNP-2008-CORRESPONDENCE(27-11-2009).pdf

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1858-MUMNP-2008-CORRESPONDENCE(30-7-2010).pdf

1858-mumnp-2008-correspondence.pdf

1858-mumnp-2008-description(complete).doc

1858-mumnp-2008-description(complete).pdf

1858-mumnp-2008-drawing.pdf

1858-MUMNP-2008-EP DOCUMENT(10-4-2012).pdf

1858-MUMNP-2008-FORM 1(28-8-2008).pdf

1858-mumnp-2008-form 1.pdf

1858-MUMNP-2008-FORM 13(7-2-2012).pdf

1858-MUMNP-2008-FORM 18(27-11-2009).pdf

1858-mumnp-2008-form 2(title page).pdf

1858-mumnp-2008-form 2.doc

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1858-MUMNP-2008-FORM 3(10-12-2013).pdf

1858-MUMNP-2008-FORM 3(12-8-2011).pdf

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1858-MUMNP-2008-FORM 3(15-2-2012).pdf

1858-MUMNP-2008-FORM 3(22-6-2013).pdf

1858-MUMNP-2008-FORM 3(23-1-2013).pdf

1858-MUMNP-2008-FORM 3(24-2-2010).pdf

1858-MUMNP-2008-FORM 3(28-8-2008).pdf

1858-MUMNP-2008-FORM 3(4-8-2010).pdf

1858-MUMNP-2008-FORM 3(8-8-2012).pdf

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1858-mumnp-2008-pct-ipea-409.pdf

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1858-mumnp-2008-pct-isa-210.pdf

1858-MUMNP-2008-REPLY TO EXAMINATION REPORT(10-4-2012).pdf

1858-MUMNP-2008-REPLY TO EXAMINATION REPORT(14-1-2013).pdf

1858-MUMNP-2008-REPLY TO EXAMINATION REPORT(22-6-2012).pdf

1858-MUMNP-2008-US DOCUMENT(10-4-2012).pdf

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