Liquid soap having enhanced antibacterial activity

Abstract

Method of cleansing human skin employing an aqueous soap composition comprising of fatty acid soap, at least one silver (I) compound having a silver ion solubility (in water at 25? C.) of at least 1?10.sup.?4 mol/L, and water, which composition provides biocidal activity against Gram positive bacteria in particular S. aureus, in relatively short contact periods.

Claims

1. A method of reducing Gram positive bacteria comprising Staphylococcus aureus, on human skin which comprises: a) providing a liquid cleansing formulation comprising: (i) from 5 to 65% by weight, based on the total weight of the formulation, of fatty acid soaps, wherein the fatty acid soaps are derived from fatty acids comprising lauric acid and oleic acid, (ii) 0.1 to 100 ppm by weight, based on the total weight of the formulation, of at least one silver(I) compound having a silver ion solubility (in water at 25? C.) of at least 1?10.sup.?4 mol/L; and (iii) 20 to 90% by weight water; b) diluting the liquid cleansing formulation to form an aqueous soap composition having a pH in the range of 9 to 11, said aqueous soap composition comprising: (i) from 0.2 to 25% by weight of the fatty acid soaps; (ii) said at least one silver(I) compound having a silver ion solubility (in water at 25? C.) of at least 1?10.sup.?4 mol/L; c) reducing the Staphylococcus aureus present on human skin by applying to the human skin said aqueous soap composition; d) allowing the aqueous soap composition to remain on the human skin for a contact period of under 1 minute; and d) rinsing the aqueous soap composition from the human skin.

2. A method as described in claim 1 wherein the silver(I) compound is present in the liquid cleansing formulation in an amount of from 1 to 50 ppm, based on the total weight of the formulation.

3. A method as described in claim 1 wherein the silver compound comprises silver oxide.

Description

DETAILED DESCRIPTION OF THE INVENTION

(1) Except in the operating and comparative examples, or where otherwise explicitly indicated, all numbers in this description indicating amounts, parts, percentages, ratios, and proportions of material, physical properties of material, and conditions of reaction are to be understood as modified by the word about. All parts, percentages, ratios, and proportions of material referred to in this description are by weight unless otherwise indicated.

(2) The term comprising is meant not to be limiting to any subsequently stated elements but rather to encompass non-specified elements of major or minor functional importance. In other words, the listed steps, elements or options need not be exhaustive. Whenever the words including or having are used, these terms are meant to be equivalent to comprising as defined above. Where the compositions or formulations of the subject invention are described as including or comprising specific components or materials, narrower embodiments where the compositions or formulations can consist essentially of or consist of the recited components or materials are also contemplated.

(3) It should also be noted that in specifying any range of concentration or amount, any particular upper concentration or amount can be associated with any particular lower concentration or amount.

(4) Fatty Acid Soap

(5) The term fatty acid soap or, more simply, soap is used here in its popular sense, i.e., salts of aliphatic alkane- or alkene monocarboxylic fatty acids preferably having 6 to 22 carbon atoms, and preferably 8 to 18 carbon atoms.

(6) The fatty acid soap should comprise from 5 to 65% by weight, preferably from 10 to 55% by weight of the subject liquid cleansing formulations. The referenced fatty acid levels are for formulations in the form typically provided to consumers, without taking into account dilution in use. In use, formulations containing fatty acid soap at higher levels within such range are typically diluted with water such that the diluted composition that is applied to the skin typically contains 25% or less, more particularly from 0.2 to 25% or less of fatty acid soap.

(7) Typical of the soap salts are alkali metal or alkanol ammonium salts of such fatty acids, although other metal salts thereof, e.g., magnesium salts, may also be employed. Sodium, potassium, magnesium, mono-, di- and tri-ethanol ammonium salts of such acids are among the more common soaps suitable for use herein. In one or more commonly, potassium soaps are used in the formulations of this invention, but up to about 25% of the soap may be sodium or magnesium soaps.

(8) As noted above, the fatty acids from which the soap salts are derived may contain unsaturation. The level of unsaturation should be in accordance with commercially acceptable standards. Excessive unsaturation is normally avoided to minimize color and odor issues. Commonly, not more than to 40 wt. % of the fatty acids from which the soap salts are formed are unsaturated. In one more embodiments, from 10 to 40 wt. %, more particularly from 20 to 40 wt. % of the fatty acids from which the soap salts are formed are unsaturated.

(9) In one or more embodiments it is preferred that the combination of C.sub.12, C.sub.14, C.sub.16, and C.sub.18 fatty acids accounts for from 90 to 100 wt %, more particularly from 95 to 100 wt. % of the total fatty acids from which the soap salts are formed and, preferably, that the combination of C.sub.16 and C.sub.18 fatty acids accounts for from 10 to 35 wt. %, more particularly from 12 to 30 wt %, even more particularly from 14 to 28 wt. %, of the total fatty acids from which the soap salts are formed. Preferably, the total amount of C.sub.8 and C.sub.10 fatty acids is less than 5 wt %, and preferably is less than 3 wt. % of the fatty acids from which the soap salts are formed.

(10) Silver Compounds

(11) The silver compounds employed as in the subject formulations and compositions are one or more water-soluble silver (I) compounds having a silver ion solubility at least 1.0?10.sup.?4 mol/L (in water at 25? C.). Silver ion solubility, as referred to herein, is a value derived from a solubility product (Ksp) in water at 25? C., a well known parameter that is reported in numerous sources. More particularly, silver ion solubility [Ag+], a value given in mol/L may be calculated using the formula:
[Ag+]=(Ksp.Math.x).sup.(1/(x+1)),

(12) wherein Ksp is the solubility product of the compound of interest in water at 25? C., and x represents the number of moles of silver ion per mole of compound. It has been found that silver (I) compounds having a silver ion solubility of at least 1?10.sup.?4 mol/L are suitable for use herein. Silver ion solubility values for a variety of silver compounds are given in Table 1:

(13) TABLE-US-00001 TABLE 1 Silver ion Solubility Values Silver Ion Ksp Solubility [Ag+] (mol/L in water (mol/L in water Silver Compound X at 25? C.) at 25? C.). silver nitrate 1 51.6 7.2 Silver acetate 1 2.0 ? 10.sup.?3 4.5 ? 10.sup.?2 Silver sulfate 2 1.4 ? 10.sup.?5 3.0 ? 10.sup.?2 Silver benzoate 1 2.5 ? 10.sup.?5 5.0 ? 10.sup.?3 Silver salicylate 1 1.5 ? 10.sup.?5 3.9 ? 10.sup.?3 Silver carbonate 2 8.5 ? 10.sup.?12 2.6 ? 10.sup.?4 Silver citrate 3 2.5 ? 10.sup.?16 1.7 ? 10.sup.?4 Silver oxide 1 2.1 ? 10.sup.?8 1.4 ? 10.sup.?4 Silver phosphate 3 8.9 ? 10.sup.?17 1.3 ? 10.sup.?4 Silver chloride 1 1.8 ? 10.sup.?10 1.3 ? 10.sup.?5 Silver bromide 1 5.3 ? 10.sup.?13 7.3 ? 10.sup.?7 Silver iodide 1 8.3 ? 10.sup.?17 9.1 ? 10.sup.?9 Silver sulfide 2 8.0 ? 10.sup.?51 .sup.2.5 ? 10.sup.?17

(14) Among the silver (I) compounds suitable for use herein are silver oxide, silver nitrate, silver acetate, silver sulfate, silver benzoate, silver salicylate, silver carbonate, silver citrate and silver phosphate, with silver oxide, silver sulfate and silver citrate being of particular interest in one or more embodiments. In at least one preferred embodiment the silver (I) compound comprises silver oxide.

(15) In at least one embodiment the silver compound is not silver dihydrogen citrate. In another embodiment the silver compound is not a salt of alginic acid or substantially depolymerized alginic acid. In one preferred embodiment the silver compound is not in the form of nano particles, attached to nano particles or part of intercalated silicates such as, for example, bentonite.

(16) In one or more embodiments, the ratio by weight of silver compound to soap salt in the aqueous soap compositions employed in the subject methods is from 1:500 to 1:650,000, preferably from 1:1000 to 1:250,000, more preferably from 1:1200 to 1:200,000.

(17) Water

(18) The personal cleansing formulations of the subject invention typically contain water in an amount of from 20 to 90%, by weight more particularly from 50 to 85% by weight, based on the total weight of the formulation. Such water contents are representative of a relatively broad range of formulations, including both concentrated and non-concentrates products, with formulations having water contents of from 20 to less than 50% by weight of water being typical of concentrated products.

(19) In use, the formulations are commonly diluted with water. The extent of dilution depends on the particular product form. Less commonly, but also contemplated for use herein are formulations that are foamed without dilution, typically through the use of pump dispensers in which product is passed through a screen in the pump.

(20) In a preferred embodiment, the silver-containing compositions of the invention are used in combination with essential oil antimicrobial actives thymol and terpineol. Preferably, each is present in overall silver-containing composition at level of 0.01 to 2% of the composition.

(21) Thymol is preferably present in 0.02 to 0.5%, more preferably up to 0.3% by weight, further more preferably up to 0.2% by weight of the composition. Thymol may be added to the antimicrobial composition in purified form. Alternatively, thyme oil or thyme extract comprising thymol may be added to the antimicrobial composition, while 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. Thyme plant refers to a plant belonging be genus Thymus and includes but is not limited 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.

(22) The structures of thymol and its isomer carvacrol are given below:

(23) ##STR00001##

(24) Terpineol is preferably present at 0.05 to 1% more preferably up to 0.5% by weight of the composition. The terpineol is preferably selected from alpha-terpineol, beta-terpineol, gamma-terpineol or mixtures thereof. It is particularly preferred that the terpineol is alpha-terpineol. Terpineol may be added to the antimicrobial composition in purified form. Alternatively pine oil comprising terpineol may be added to the antimicrobial composition.

(25) The structure of a terpineol compound is given below:

(26) ##STR00002##

(27) Optional Ingredients

(28) If desired, the formulations may optionally include a detersive surfactant in addition to the fatty acid soap. Such detersive surfactants include, for example, anionic, zwitterionic and/or nonionic surfactants.

(29) Examples of anionic surfactants suitable for use herein include, but are not limited to, ammonium lauryl sulfate, ammonium laureth sulfate, triethylamine lauryl sulfate, triethylamine laureth sulfate, triethanolamine lauryl sulfate, triethanolamine laureth sulfate, monoethanolamine lauryl sulfate, monoethanolamine laureth sulfate, diethanolamine lauryl sulfate, diethanolamine laureth sulfate, lauric monoglyceride sodium sulfate, sodium lauryl sulfate, sodium laureth sulfate, potassium laureth sulfate, sodium lauryl sarcosinate, sodium lauroyl sarcosinate, potassium lauryl sulfate, sodium trideceth sulfate, sodium methyl lauroyl taurate, sodium lauroyl isethionate, sodium laureth sulfosuccinate, sodium lauroyl sulfosuccinate, sodium tridecyl benzene sulfonate, sodium dodecyl benzene sulfonate, sodium lauryl amphoacetate and mixtures thereof.

(30) The anionic surfactant may be, for example, an aliphatic sulfonate, such as a primary C.sub.8-C.sub.22 alkane sulfonate, primary C.sub.8-C.sub.22 alkane disulfonate, C.sub.8-C.sub.22 alkene sulfonate, C.sub.8-C.sub.22 hydroxyalkane sulfonate or alkyl glyceryl ether sulfonate.

(31) Zwitterionic surfactants suitable for use herein include, but are not limited to derivatives of aliphatic quaternary ammonium, phosphonium, and sulfonium compounds, in which the aliphatic radicals can be straight or branched chain, and wherein one of the aliphatic substituents contains from about 8 to about 18 carbon atoms and one substituent contains an anionic group, e.g., carboxy, sulfonate, sulfate, phosphate, or phosphonate. Illustrative zwitterionic surfactants are coco dimethyl carboxymethyl betaine, cocoamidopropyl betaine, cocobetaine, oleyl betaine, cetyl dimethyl carboxymethyl betaine, lauryl bis-(2-hydroxyethyl)carboxymethyl betaine, stearyl bis-(2-hydroxypropyl)carboxymethyl betaine, oleyl dimethyl gamma-carboxypropyl betaine, lauryl bis-(2-hydroxypropyl)alpha-carboxyethyl betaine, and mixtures thereof. The sulfobetaines may include stearyl dimethyl sulfopropyl betaine, lauryl dimethyl sulfoethyl betaine, lauryl bis-(2-hydroxyethyl) sulfopropyl betaine and mixtures thereof.

(32) Nonionic surfactants which may be used include the reaction products of compounds having a hydrophobic group and a reactive hydrogen atom. Exemplative are alcohols, acids, amides or alkyl phenols reacted with alkylene oxides, especially ethylene oxide either alone or with propylene oxide. Specific nonionics are C.sub.6-C.sub.22 alkyl phenols-ethylene oxide condensates, the condensation products of C.sub.8-C.sub.18 aliphatic primary or secondary linear or branched alcohols with ethylene oxide, and products made by condensation of ethylene oxide with the reaction products of propylene oxide and ethylenediamine. Other nonionics include long chain tertiary amine oxides, long chain tertiary phosphine oxides and dialkyl sulphoxides. Also useful are the alkyl polysaccharides.

(33) When present the amount of additional detersive surfactant depends, in part, on the selection thereof, and the amount of the fatty acid soap. The subject personal cleansing formulations are soap-based, i.e., the total amount of fatty acid soap present therein exceeds the total amount of detersive surfactant that is not fatty acid soap. Commonly, the fatty acid soap comprises from 85 to 100 wt. %, more particularly from 90 to 100 wt. % of the total detersive surfactant present in the personal cleansing formulations.

(34) The formulations typically include one or more skin benefit agents. The term skin benefit agent is defined as a substance which softens or improves the elasticity, appearance, and youthfulness of the skin (stratum corneum) by either increasing its water content, adding, or replacing lipids and other skin nutrients, or both, and keeps it soft by retarding the decrease of its water content. Included among the suitable skin benefit agents are emollients, including, for example, hydrophobic emollients, hydrophilic emollients, or blends thereof.

(35) Useful skin benefit agents include the following: (a) silicone oils and modifications thereof such as linear and cyclic polydimethylsiloxanes; amino, alkyl, alkylaryl, and aryl silicone oils; (b) fats and oils including natural fats and oils such as jojoba, soybean, sunflower, rice bran, avocado, almond, olive, sesame, persic, castor, coconut, and mink oils; cacao fat; beef tallow and lard; hardened oils obtained by hydrogenating the aforementioned oils; and synthetic mono, di and triglycerides such as myristic acid glyceride and 2-ethylhexanoic acid glyceride; (c) waxes such as carnauba, spermaceti, beeswax, lanolin, and derivatives thereof; (d) hydrophobic and hydrophilic plant extracts; (e) hydrocarbons such as liquid paraffin, petrolatum, microcrystalline wax, ceresin, squalene, pristan and mineral oil; (f) higher fatty acids such as lauric, myristic, palmitic, stearic, behenic, oleic, linoleic, linolenic, lanolic, isostearic, arachidonic and poly unsaturated fatty acids (PUFA); (g) higher alcohols such as lauryl, cetyl, stearyl, oleyl, behenyl, cholesterol and 2-hexydecanol alcohol; (h) esters such as cetyl octanoate, myristyl lactate, cetyl lactate, isopropyl myristate, myristyl myristate, isopropyl palmitate, isopropyl adipate, butyl stearate, decyl oleate, cholesterol isostearate, glycerol monostearate, glycerol monolaurate, glycerol distearate, glycerol tristearate, alkyl lactate, alkyl citrate and alkyl tartrate; (i) essential oils and extracts thereof such as mentha, jasmine, camphor, white cedar, bitter orange peel, ryu, turpentine, cinnamon, bergamot, citrus unshiu, calamus, pine, lavender, bay, clove, hiba, eucalyptus, lemon, starflower, thyme, peppermint, rose, sage, sesame, ginger, basil, juniper, lemon grass, rosemary, rosewood, avocado, grape, grapeseed, myrrh, cucumber, watercress, calendula, elder flower, geranium, linden blossom, amaranth, seaweed, ginko, ginseng, carrot, guarana, tea tree, jojoba, comfrey, oatmeal, cocoa, neroli, vanilla, green tea, penny royal, aloe vera, menthol, cineole, eugenol, citral, citronelle, borneol, linalool, geraniol, evening primrose, camphor, thymol, spirantol, penene, limonene and terpenoid oils; (j) polyhydric alcohols, for example, glycerine, sorbitol, propylene glycol, and the like; and polyols such as the polyethylene glycols, examples of which are: Polyox WSR-205 PEG 14M, Polyox WSR-N-60K PEG 45M, or Polyox WSR-N-750, and PEG 7M; (k) lipids such as cholesterol, ceramides, sucrose esters and pseudo-ceramides as described in European Patent Specification No. 556,957; (I) vitamins, minerals, and skin nutrients such as milk, vitamins A, E, and K; vitamin alkyl esters, including vitamin C alkyl esters; magnesium, calcium, copper, zinc and other metallic components; (m) sunscreens such as octyl methoxyl cinnamate (Parsol MCX) and butyl methoxy benzoylmethane (Parsol 1789); (n) phospholipids; and (o) anti-aging compounds such as alpha-hydroxy acids and beta-hydroxy acids. Skin benefit agents commonly account for up to 30 wt. % of the liquid soap formulation, with levels of from 0 to 25 wt. %, more particularly from 0 to 20 wt %, being typical of the levels at which those skin benefit agents generally known as emollients are employed in many of the subject formulations. Preferred skin benefit agents include fatty acids, hydrocarbons, polyhydric alcohols, polyols and mixtures thereof, with emollients that include at least one C.sub.12 to C.sub.18 fatty acid, petrolatum, glycerol, sorbitol and/or propylene glycol being of particular interest in one or more embodiments.

(36) Other optional ingredients include water soluble/dispersible polymers. These polymers can be cationic, anionic, amphoteric or nonionic types with molecular weights higher than 100,000 Dalton. They are known to increase the viscosity and stability of liquid personal cleansing formulation, to enhance in-use and after-use skin sensory properties, and to enhance lather creaminess and lather stability. When present, the total amount of such polymers commonly ranges from 0.1 to 10% by weight of the personal cleansing formulation.

(37) Examples of water soluble or dispersible polymers include the carbohydrate gums such as cellulose gum, microcrystalline cellulose, cellulose gel, hydroxyethyl cellulose, hydroxypropyl cellulose, sodium carboxymethylcellulose, methyl cellulose, ethyl cellulose, guar gum, gum karaya, gum tragacanth, gum arabic, gum acacia, gum agar, xanthan gum and mixtures thereof; modified and nonmodified starch granules and pregelatinized cold water soluble starch; emulsion polymers such as Aculyn? 28, Aculyn? 22 or Carbopol? Aqua SF1; cationic polymer such as modified polysaccharides including cationic guar available from Rhone Poulenc under the trade name Jaguar C13S, Jaguar C14S, Jaguar C17, or Jaguar C16; cationic modified cellulose such as UCARE Polymer JR 30 or JR 40 from Amerchol; N-Hance? 3000, N-Hance? 3196, N-Hance? GPX 215 or N-Hance? GPX 196 from Hercules; synthetic cationic polymer such as Merquat? 100, Merquat? 280, Merquat? 281 and Merquat? 550 sold by Nalco; cationic starches such as StaLok? 100, 200, 300 and 400 sold by Staley Inc.; cationic galactomannans such as Galactasol? 800 series by Henkel, Inc.; Quadrosoft? LM-200; and Polyquaternium-24. Also suitable are high molecular weight polyethylene glycols such as Polyox? WSR-205 (PEG 14M), Polyox? WSR-N-60K (PEG 45), and Polyox? WSR-301 (PEG 90M).

(38) Preservatives/antimicrobials can desirably be incorporated into the personal cleansing formulations of this invention to protect against the growth of potentially harmful microorganisms. Suitable traditional preservatives for formulations of this invention are alkyl esters of para-hydroxybenzoic acid. Other preservatives/antimicrobials which have more recently come into use include hydantoin derivatives, propionate salts, and a variety of quaternary ammonium compounds. Among the preservatives/antimicrobials of particular interest are phenoxyethanol, methyl paraben, propyl paraben, imidazolidinyl urea, sodium dehydroacetate and benzyl alcohol. Other preservatives of particular interest are dimethyloldimethylhydantoin (Glydant XL1000), parabens, sorbic acid, thymol and terpineol to name a few (with combinations of thymol and terpineol as described, for example, in U.S. Patent Application Publication No. 2011/0223114 incorporated herein by reference, being of particular interest in one or more embodiments).

(39) The preservatives/antimicrobials should be selected having regard for the use of the formulation and possible incompatibilities between the preservatives and other ingredients. Preservatives are preferably employed in amounts ranging from 0.01 to 2% by weight of the personal cleansing formulation.

(40) Additional optional ingredients which may be present in the subject personal cleansing formulations are, for example: fragrances; sequestering and chelating agents such as tetrasodium ethylenediaminetetraacetate (EDTA), ethane hydroxyl diphosphonate (EHDP), and etidronic acid, aka 1-hydroxyethylidene diphosphonic acid (HEDP); coloring agents; opacifiers and pearlizers such as zinc stearate, magnesium stearate, TiO.sub.2, ethylene glycol monostearate (EGMS), ethylene glycol distearate (EGDS) or Lytron 621 (Styrene/Acrylate copolymer) and the like; pH adjusters; antioxidants, for example, butylated hydroxytoluene (BHT) and the like; stabilizers; suds boosters, such as for example, coconut acyl mono- or diethanol amides; ionizing salts, such as, for example, sodium chloride and sodium sulfate, and other ingredients such as are conventionally used in liquid soap formulations. The total amount of such additional optional ingredients is typically from 0 to 10% by weight, more particularly from 0.1 to 5% by weight, based on the total weight of the personal cleansing formulation.

(41) The personal cleansing formulations of this invention are of interest with respect to biocidal activity against Gram positive bacteria, including in particular S. aureus. Other Gram, positive bacteria against which the soap formulations are of interest are S. epidermidis, and/or Corynebacteria, in particular, Corynebacteria strains responsible for the hydrolysis of axilla secretions to malodorous compounds. Desirably, the formulations provides a log.sub.10 reduction in biocidal activity against Staphylococcus aureus ATCC 6538 of at least 2.5, preferably of least 3, at a contact time of 30 seconds and even more preferably provides a Log.sub.10 Reduction against S. aureus ATCC 6538 of at least 1.5 even more preferably at least 2 at a contact time of 10 seconds.

(42) In use, the personal cleansing formulations are diluted, as needed, to form aqueous cleansing compositions that are applied to the skin for contact times less than 1 minute, more particularly 30 seconds or less (with contact times of 10 to 30 seconds being of interest with respect to contact times of a moderate to a relatively long duration and contact times of 10 seconds or less being of interest with respect to contact times of short to moderate duration) and thereafter is removed from the skin, typically by rinsing with water. The personal cleansing formulation can be diluted before, after or simultaneous with its being placed on the skin, with dilution typically occurring by the formulation being worked into a lather in the hands or on an applicator, such as a facecloth, sponge or pouf.

(43) Manufacture

(44) The personal cleansing formulations herein described may be produced by preparative techniques as are conventional in the art. In one very general methodology, to a heated aqueous phase of water, is added melted fatty acid, followed by caustic (to neutralize the fatty acid and form soap), synthetic detergents and co-solvents; the remaining ingredients are added, as appropriate, as the product is cooled to room temperature.

(45) The personal cleansing formulations may be provided in a variety of different product forms including, for example, hand, face and body washes, shower gels, and the like. The formulations may be provided in bottles, pump dispensers, tubes, sachets, or other packaging suitable for the product form.

EXAMPLES

(46) The following non-limiting examples are provided to further illustrate the invention; the invention is not in any way limited thereto. The following protocol was used to evaluate biocidal activity.

(47) In-Vitro Time-Kill Protocol

(48) Soap Solution Preparation

(49) Solution preparation depends, in part, on the particular form of the liquid soap formulation. For example, formulations that are not diluted in use, e.g., self-foaming formulations, are employed as is. A formulation that contains 30 wt. % or less of detersive surfactant and which is intended to be diluted in use is mixed with an equal amount, by weight, of water to form a soap solution containing 50 wt % of the initial formulation. A formulation that contains more than 30 wt % of detersive surfactant and which is intended to be diluted in use is mixed with water to form a soap solution containing 16 wt % of the initial formulation.

(50) Bacteria

(51) Staphylococcus aureus ATCC 6538, were used in this study to represent Gram positive bacteria, respectively. The bacteria was stored at ?80? C. Fresh isolates were cultured twice on Tryptic Soy Agar plates for 24 hours at 37? C. before each experiment.

(52) In-Vitro Time-Kill Assay

(53) Time-kill assays are performed according to the European Standard, EN 1040:2005 entitled Chemical Disinfectants and AntisepticsQuantitative Suspension Test for the Evaluation of Basic Bactericidal Activity of Chemical Disinfectants and AntisepticsTest Method and Requirements (Phase 1) incorporated herein by reference Following this procedure Growth-phase bacterial cultures at 1.5?10.sup.8 to 5?10.sup.8 colony forming units per ml (cfu/ml) are treated with the soap solutions (prepared as described above) at 25? C. In forming the test sample 8 parts by weight of the soap solution, prepared as described above, is combined with 1 part by weight of culture and 1 part by weight of water. After 10, 30, and 60 seconds of exposure, samples are neutralized to arrest the antibacterial activity of the soap solutions. Then test solutions are serially diluted, plated on solid medium, incubated for 24 hours and surviving cells are enumerated. Bactericidal activity is defined as the log reduction in cfu/ml relative to the bacterial concentration at 0 seconds. Cultures not exposed to any soap solutions serve as no-treatment controls.

(54) The log.sub.10 reduction was calculated using the formula:
Log.sub.10 Reduction=log.sub.10 (numbers control)?log.sub.10 (test sample survivors)

Examples 1 to 3

(55) Liquid soap formulations were prepared as indicated in Table 2 below.

(56) TABLE-US-00002 TABLE 2 Ingredient Liquid Liquid Liquid (wt. %) Soap 1 Soap 2 Soap 3 Water Balance Balance Balance to 100 To 100 To 100 Cellulose Ether 0.30 0.30 0.30 (Methocel? 40-100 from Dow Chemical) BHT 0.05 0.05 0.05 EDTA 0.13 0.13 0.13 EHDP 0.08 0.08 0.08 Glycerine 0.50 0.50 0.50 Laurie Acid Soap 5.8 5.8 5.8 Myristic Acid Soap 6.7 6.7 6.7 Palmitic Acid Soap 2.1 2.1 2.1 KOH 3.60 3.60 3.60 Sodium laureth ether 2.10 2.10 2.10 Sulfate-1EO (SLES) Cocamidopropyl Betaine 0.75 0.75 0.75 Methylchloroisothiozolinone 0.10 0.10 0.10 (Kathon? CG) Silver Sulfate 0.0001 0.0005

(57) The biocidal activity of the liquid soaps formulations was evaluated following the above described In-Vitro Time-Kill Protocol. Biocidal activity results are reported in Table 3.

(58) TABLE-US-00003 TABLE 3 Biocidal Activity Log.sub.10 Reduction against S. aureus ATCC 6538 Contact Time 10 Seconds 30 Seconds 60 Seconds Liquid Soap 1 (Comp.) 1.0 2.0 2.9 Liquid Soap 2 1.5 2.8 4.3 Liquid Soap 3 2.5 3.4 4.1

(59) As demonstrated by the Table 3 data, at the indicated contact times, liquid soap 2 and liquid soap 3 had greater bactericidal efficacy against S. aureus ATCC 6538 than liquid soap 1 (no silver component).