Foaming Topical Antimicrobial Cleaning Compositions

Abstract

Provided are largely aqueous topical antimicrobial cleaning composition which provides an antimicrobial benefit, as well as methods for their production and methods for their use. The compositions comprise at least two different cationic surfactants which provide a primary antimicrobial benefit, and as a predominant further surfactant an amine oxide surfactant. The composition are at an acidic pH and are highly effective in providing an antimicrobial benefit to topical surfaces, e.g, epidermis and hair.

Claims

1. A highly aqueous, antimicrobially effective topical composition which comprises: at least one first cationic surfactant or salt thereof having antimicrobial or germicidal properties or salt form thereof, at least one second quaternary ammonium cationic surfactant or salt form thereof having antimicrobial or germicidal properties according to the structure (I) ##STR00008## wherein: R.sub.1 is a C.sub.20-C.sub.36, preferably an unsubstituted alkyl moiety, R.sub.2, R.sub.3 and R.sub.4 are each independently C.sub.1-C.sub.3 alkyl moieties, and, X is a salt forming counterion which renders the second quaternary surfactant water soluble or water dispersible, wherein the said second quaternary ammonium cationic surfactant or salt form thereof is different from the said first cationic surfactant or salt form thereof, and, an amine oxide nonionic surfactant which is the predominant further surfactant in the composition, at least 75% wt. water; wherein the compositions are in the pH range of 4.5-6, and which compositions are low viscosity, pourable unpressurized liquids.

2. A composition according to claim 1, wherein: the amine oxide surfactant comprises at least 90%, of the total weight of all surfactants present in the composition other than the amounts of the at least one first cationic surfactant having antimicrobial or germicidal properties or salt form thereof and with the at least one different, second quaternary ammonium cationic surfactant or salt form thereof.

3. (canceled)

4. A composition according to claim 1, wherein the amine oxide surfactant is the sole further surfactants present in composition other than the amounts of the at least one first cationic surfactant having antimicrobial or germicidal properties or salt form thereof and with the at least one different, second quaternary ammonium cationic surfactant or salt form thereof.

5. A composition according to claim 1, wherein the compositions are in the pH range of 4.5-5.5.

6. A composition according to claim 1, wherein the composition further comprises: glycerin, a glycol, and a oxyalkylenated compound.

7. A composition according to claim 6 wherein the oxyalkylenated compound is a polyethylene glycol having from 100 to 200 ethylene oxide units.

8. A composition according to claim 1, wherein the said composition exhibits at least a 3log.sub.10 reduction (99.9%), of one or more of the challenge microorganisms selected from one or more, preferably at least two or more, ore preferably at least 3 or more of the following microorganisms: E. coli, S. aureus, P. aeruginosa, and E. hirae according to EN 1276.

9. A composition according to claim 8, wherein the said composition exhibits at least a 4log.sub.10 reduction of one or more the following challenge microorganisms: Enterococcus faecalis (ATCC #51299) Esherichia coli (ATCC #11229) Klebsiella pneumoniae pneumoniae (ATCC #10031) Psuedomonas aeruginosa (ATCC #9027) Staphylococcus aureus aureus (ATCC #6538) Staphylococcus aureus aureus MRSA (ATCC #33592) Staphylococcus epidermidis (ATCC #12228) Streptococcus pneumoniae (ATCC #6303) Streptococcus pyogenes (ATCC #19615) when tested according to ASTM E-1054-08, Standard Test Methods for Evaluation of Inactivators of Antimicrobial Agents for a 60 second contact time.

10. (canceled)

12. A composition according to claim 1, which comprises at least 90% wt. of water.

13. A composition according to claim 3, which comprises at least 90% wt. of water.

14. A highly aqueous, antimicrobially effective topical composition according to claim 1 which comprises: up to 2% wt. of at least one first cationic surfactant or salt thereof having antimicrobial or germicidal properties or salt form thereof, up to 10% wt. of at least one, second quaternary ammonium cationic surfactant or salt form thereof having antimicrobial or germicidal properties according to the structure (I) ##STR00009## wherein: R.sub.1 is a C.sub.20-C.sub.36, preferably an unsubstituted alkyl moiety, R.sub.2, R.sub.3 and R.sub.4 are each independently C.sub.1-C.sub.3 alkyl moieties, and, X is a salt forming counterion which renders the second quaternary surfactant water soluble or water dispersible, wherein the said second quaternary ammonium cationic surfactant or salt form thereof is different from the said first cationic surfactant or salt form thereof, and, up to 15% wt. of an amine oxide nonionic surfactant and which amine oxide nonionic surfactant is the predominant further surfactant in the composition comprising at least 80% wt. of the total weight of all surfactants present in the largely aqueous topical antimicrobial cleaning composition other than the amounts of the at least one first cationic surfactant having antimicrobial or germicidal properties or salt form thereof and with the at least one different, second quaternary ammonium cationic surfactant or salt form thereof, at least 75% wt. water; wherein the compositions are in the pH range of 4.5-5.5, and which compositions are low viscosity, pourable unpressurized liquids.

15. A highly aqueous, antimicrobially effective topical composition according to claim 14, which composition further comprises glycerin, a glycol, and a oxyalkylenated compound.

16. A highly aqueous, antimicrobially effective topical composition according to claim 1, wherein the composition exhibits antimicrobial efficacy against one or more of the following challenge microorganisms: Acinetobacter baumannii (ATCC #19606) Acinetobacter lwoffii (ATCC #15309) Burkholderia cepacia (ATCC #25416) Candida albicans (ATCC #10231) Corynebacterium jeikeium (ATCC #43734) Enterobacter aerogenes (ATCC #13048), Enterobacter cloacae cloacae (ATCC #13047), Enterococcus faecalis (ATCC #19433) Enterococcus faecalis VRE (ATCC #51299) Enterococcus faecium (ATCC #19434) Escherichia coli (ATCC #11229) Escherichia coli (ATCC #25922) Haemophilus influenzae (ATCC #19418) Klebsiella oxytoca (ATCC #43165) Klebsiella pneumoniae ozaenae (ATCC #11296) Micrococcus luteus (ATCC #4698) Proteus mirabilis (ATCC #7002) Pseudomonas aeruginosa (ATCC #9027) Serratia marcescens (ATCC #14756) Staphylococcus aureus aureus (ATCC #6538) Staphylococcus aureus aureus MRSA (ATCC #33592) Staphylococcus epidermidis (ATCC #12228) Staphylococcus haemolyticus (ATCC #29970) Staphylococcus hominis hominis (ATCC #27844) Staphylococcus saprophyticus (ATCC #15305) Streptococcus pneumoniae (ATCC #6303) Streptococcus pyogenes (ATCC #19615) MRSA=Methicillin-Resistant Staphylococcus aureus VRE=Vancomycin-Resistant Enterococcus.

17. An aqueous, antimicrobially effective topical composition comprising: at least one first cationic surfactant or salt thereof having antimicrobial or germicidal properties; at least one second cationic surfactant having antimicrobial or germicidal properties which is a quaternary ammonium surfactant having the structure (I) ##STR00010## wherein: R.sub.1 is an unsubstituted C.sub.16, or a C.sub.20-C.sub.36 alkyl moiety, R.sub.2, R.sub.3 and R.sub.4 are each independently C.sub.1-C.sub.3 alkyl moieties, and, X is a salt forming counterion which renders the surfactant water soluble or water dispersible, wherein the second cationic surfactant is different from the first cationic surfactant or salt thereof, and wherein the weight ratios of the at least one second cationic surfactant:at least one cationic surfactant is from 9:1 to 34.6:1, and, an amine oxide nonionic surfactant which, in weight percent, is the predominant further surfactant in the composition, glycerin; a glycol; a polyethylene glycol having from 100 to 200 ethylene oxide units, and, at least 75% wt. water; wherein the composition has a pH in the range of 4.5-5.5, and the composition is a pourable, unpressurized liquid; wherein benzophenones are excluded from the composition, and further wherein the composition exhibits at least a 3log.sub.10 reduction (99.9%) according to EN 1276 of one or more of challenge microorganisms selected from: E. coli, S. aureus, P. aeruginosa, and E. hirae.

18. The composition according to claim 17, wherein: the amine oxide surfactant comprises at least 95% of the total weight of all surfactants present in the composition exclusive of the said at least one first cationic surfactant or salt thereof and the said at least one second quaternary ammonium cationic surfactant.

19. The composition according to claim 17, wherein said composition exhibits at least a 4log.sub.10 reduction of one or more the following challenge microorganisms: Enterococcus faecalis (ATCC #51299) Esherichia coli (ATCC #11229) Klebsiella pneumoniae pneumoniae (ATCC #10031) Psuedomonas aeruginosa (ATCC #9027) Staphylococcus aureus aureus (ATCC #6538) Staphylococcus aureus aureus MRSA (ATCC #33592) Staphylococcus epidermidis (ATCC #12228) Streptococcus pneumoniae (ATCC #6303) Streptococcus pyogenes (ATCC #19615) when tested according to ASTM E-1054-08, Standard Test Methods for Evaluation of Inactivators of Antimicrobial Agents for a 60 second contact time.

20. The composition according to claim 17, which comprises at least 90% wt. of water.

21. The composition according to claim 17, wherein the composition exhibits at least a 3log.sub.10 reduction according to EN 1276 of at least three challenge microorganisms selected from: E. coli, S. aureus, P. aeruginosa, and E. hirae.

22. A topical composition which exhibits efficacy against one or both of Methicillin-Resistant Staphylococcus aureus and Vancomycin-Resistant Enterococcus which comprises: up to 2% wt. of at least one first cationic surfactant or salt thereof having antimicrobial or germicidal properties; up to 10% wt. of at least one, second cationic surfactant which is a quaternary ammonium surfactant having antimicrobial or germicidal properties having the structure (I) ##STR00011## wherein: R.sub.1 is a an unsubstituted C.sub.16, or C.sub.20-C.sub.36 alkyl moiety, R.sub.2, R.sub.3 and R.sub.4 are each independently C.sub.1-C.sub.3 alkyl moieties, and, X is a salt forming counterion which renders the surfactant water soluble or water dispersible, and wherein the said second cationic surfactant is different from the said first cationic surfactant or salt thereof, and wherein the weight ratios of the at least one second cationic surfactant:at least one cationic surfactant is from 9:1 to 34.6:1, and, up to 15% wt. of an amine oxide nonionic surfactant which surfactant is the predominant further surfactant in the composition comprising at least 80% wt. of the total weight of all surfactants present in the composition other than the amounts of the at least one first cationic surfactant and the at least one second cationic surfactant; glycerin, a glycol, a polyethylene glycol having from 100 to 200 ethylene oxide units, and, at least 75% wt. water; wherein the composition has a pH in the range of about 4.5 to about 5.5 and, the composition is a pourable unpressurized liquid; and, wherein benzophenones are excluded from the composition.

Description

EXAMPLES

[0123] A number of largely aqueous topical antimicrobial cleaning compositions were produced according to the process described below, are described on Table 1 below. In the following compositions, the constituents were used as supplied from their respective suppliers and may constitute less than 100% wt. actives, or may have been supplied as constituting 100% wt. active of the named compound, as indicated in the following Tables 1 and 2. The amounts indicated on Table 1 refer to % wt. of the as supplied named constituent used in a composition, as supplied by the corresponding constituent listed on Table 2. To each of the compositions was included deionized water in quantum sufficient (q.s.) in order to provide 100 parts by weight of the specific composition.

[0124] Compositions which are comparative examples are identified by digits preceded with the letter C, while compositions according to the invention are identified by digits preceded with the letter E.

TABLE-US-00001 TABLE 1 1755-192 1817-006 C1 C2 PEG-150 distearate (100%) lauramide DEA (73%) coconut monoethanolamide (100%) cetrimonium chloride (30%) 3.5 benzalkonium chloride (50%) 0.26 0.26 cocoamidopropyl hydroxysultaine 7.0 7.0 decyl glucoside 2.0 2.0 lauryl dimethyl amide oxide (30%) glycerin (100%) 2.0 2.0 di-PPG-2 myreth-10 adipate 0.1 0.1 propylene glycol (100%) 0.1 0.1 preservative 0.02 0.02 tetrasodium EDTA (100%) 0.2 0.2 citric acid (50%) q.s. q.s. fragrance 0.22 0.22 DI water (100%) q.s. q.s. pH 5.0 5.0 viscosity (cPs, Brookfield Type n.t.* n.t.* LV3 viscometer, recommended spindle, at 12 rpm, sample at 20-22 C.) % amine oxide/all surfactants 0% 0% (other than cationic surfactants), active weight basis n.t.*while the indicated formulations were not tested, during the production of the samples the compositions were visually observed to be water thin, having a very thin viscosity approximate to that of deionized water. citric acid was added in a quantum sufficient in order to adjust the composition to the desired pH. E1 E2 E3 E4 E5 E6 E7 E8 E9 PEG-150 distearate (100%) 0.8 1.6 1.6 1.6 1.4 1.0 1.0 1.0 1.0 lauramide diethanolamide (73%) 0.8 1.6 1.6 coconut monoethanolamide (100%) 0.1 0.1 0.1 0.1 0.1 cetrimonium chloride (30%) 5.0 2.5 2.5 7.5 3.0 3.0 3.0 3.0 3.0 benzalkonium chloride (50%) 0.13 0.13 0.13 0.13 0.2 0.2 0.2 0.2 0.2 lauryl dimethyl amide oxide (30%) 5.0 10 10 10 5.0 5.0 5.0 5.0 5.0 glycerin (100%) 2.0 2.0 2.0 2.0 2.0 propylene glycol (100%) 0.5 0.5 0.5 0.5 0.5 preservative 0.02 0.02 0.02 0.02 0.02 0.02 tetrasodium EDTA (100%) 0.15 0.3 0.3 0.2 0.2 0.2 0.2 0.2 citric acid (50%) 0.45 0.23 0.23 0.23 0.23 fragrance 0.22 0.30 0.40 0.40 0.20 colorant 0.0006 0.0005 0.0005 0.004 di water (100%) q.s. q.s. q.s. q.s. q.s. q.s. q.s. q.s. q.s. pH 4.79 4.82 4.82 4.81 5.36 4.8-5.5 4.8-5.5 4.8-5.5 4.8-5.5 viscosity (cPs, Brookfield Type n.t.* n.t * n.t.* n.t.* 5-50 5-50 5-50 5-50 5-50 LV3 viscometer, recommended spindle (#1), at 60 rpm, sample at 20-22 C.) % amine oxide/all surfactants 72.1% 100%.sup. 71.9% 71.9% 96.7% 93.7% 93.7% 93.7% 93.7% (other than cationic surfactants), active weight basis n.t.*while the indicated formulations were not quantitatively tested, during the production of the samples the compositions were visually observed to be water thin, having a very thin viscosity which was approximate to that of deionized water. E10 E11 E12 E13 PEG-150 distearate (100%) 1.0 1.0 1.0 1.0 lauramide diethanolamide (73%) coconut monoethanolamide (100%) 0.1 0.1 0.1 0.1 cetrimonium chloride (30%) 3.0 3.0 3.0 3.0 benzalkonium chloride (50%) 0.2 0.2 0.2 0.2 lauryl dimethyl amide oxide (30%) 5.0 5.0 5.0 5.0 glycerin (100%) 2.0 2.0 2.0 2.0 propylene glycol (100%) 0.5 0.5 0.5 0.5 preservative 0.02 0.02 0.02 0.02 tetrasodium EDTA (100%) 0.2 0.2 0.2 0.2 citric acid (50%) 0.23 0.23 0.23 0.23 fragrance** 0.30 0.40 0.40 0.20 colorant 0.00062 0.00058 0.00057 0.00040 di water (100%) q.s. q.s. q.s. q.s. pH 4.8-5.5 4.8-5.5 4.8-5.5 4.8-5.5 viscosity (cPs, Brookfield Type 5-50 5-50 5-50 5-50 LV3 viscometer, recommended spindle (#1), at 60 rpm, sample at 20-22 C.) % amine oxide/all surfactants 93.7% 93.7% 93.7% 93.7% (other than cationic surfactants), active weight basis n.t*while the indicated formulations were not quantitatively tested, during the production of the samples the compositions were visually observed to be water thin, having a very thin viscosity which was approximate to that of deionized water. fragrance**the identity of the fragrance constituent used in E10-E13 was different for each of these example compositions.
The constituents used to produce the compositions of Table 1 were as follows.

TABLE-US-00002 TABLE 2 supplied by, (% wt. actives), constituent source/supplier PEG-150 distearate (100%) Hallstar PEG 6000DS (100% wt. actives) ex. Hallstar lauramide diethanolamide (73%) Ninol 55-LL (73% wt. actives) ex. coconut monoethanolamide (100%) Mackamine CMA (100% wt. actives) ex. Rhodia cetrimonium chloride (30%) Ammonyx CETAC-30 (30% wt. actives) ex. Stepan co. benzalkonium chloride (50%) Barquat MB50 (50% wt. actives) ex. Lonza lauryl dimethyl amine oxide (30%) Ammonyx LO (30% wt. actives) ex. Stepan co. glycerin (100%) glycerin (100% wt. actives) USP grade propylene glycol propylene glycol (100% wt. actives), USP grade, ex. Dow preservative Kathon CG (used as supplied) ex. Rohm & Haas tetrasodium EDTA Trilon B (100% wt. actives) ex. BASF citric acid (50%) aqueous dilution of technical grade citric acid (50% wt. actives) fragrance proprietary composition of its respective supplier colorant aqueous dispersion of one or more of: D&C Green #5, FD&C Red #33, FD&C Yellow #5, D&C Orange #4, FD&C Red #4 di water deionized water

[0125] The compositions according to the invention may be produced by simple mixing of the constituents in order to produce a homogenous composition.

[0126] A preferred method of production includes the following steps:

a) Approximately 50% of the total amount of water is charged to a beaker (or other suitable vessel) into which is provided a motorized stirrer which is activated to ask take the contents of the beaker. The water is heated to 60-65 C. after which is added (when present) the chelating agent, the PEG-150 distearate, and the alkanolamide nonionic surfactant (coconut monoethanolamide or lauramide diethanolamide) and stirring continues for approximately 10-30 minutes until the mixture within the beaker is fully heated, and is homogenous.
b) Thereafter, the heat source is removed or deactivated, and the remaining amount of water is added to the beaker on the stirring conditions in order to cool the contents of the beaker. Thereafter, are added under stirring conditions the second cationic compound (cetrimonium chloride), followed by the amine oxide surfactant, and followed by the glycerin (if present). Stirring continues until the contents of the beaker cool to approximately 30-35 C.
c) Subsequently, under stirring conditions are added the first cationic compound, any fragrance constituent (if present), followed by the remaining constituents which are added to the composition, e.g., preservatives, colorants and the like. Finally, sufficient amount of a pH adjusting agent, e.g., 50% aqueous solution of citric acid, is added in order to provide a desired product pH. Stirring continues until at least a homogenous mixture is formed, and advantageously an additional 10-30 of stirring is provided. Thereafter, the formed treatment composition can be removed from the beaker and may be used.

Antimicrobial Efficacy (I)EN1276 Test:

[0127] The antimicrobial efficacy of certain compositions of Table 1 were evaluated in accordance with the EN1276 Standard Suspension Test for bactericidal activity. Samples of each test composition were placed in test tube with 3% BSA (dirty conditions) and challenge organism (to achieve 80% v/v) for a contact time of 1 minute, followed by neutralization, serial dilutions, plating and incubation. The results of the tested compositions are reported on the following Tables 3A-3E The acceptance criteria (success criteria) for the purposes of the study was at least a 3log.sub.10 reduction (99.9%) in 60 seconds of one or more of the challenge microorganisms.

TABLE-US-00003 TABLE 3A tested composition, E1 Organism S. aureus E. coli P. aeruginosa E. hirae Inoculum Level 7.69 7.66 7.67 7.68 Replicate 1 2 1 2 1 2 1 2 Recovery <2.18 <2.18 <2.18 <2.18 <2.18 <2.18 <2.18 <2.18 Log.sub.10 >5.51 >5.51 >5.48 >5.48 >5.49 >5.49 >5.50 >5.50 Reduction Average Log.sub.10 >5.51 >5.48 >5.49 >5.50 Reduction % eradication >99.999 >99.999 >99.999 >99.999

TABLE-US-00004 TABLE 3B tested composition, E2 Organism S. aureus E. coli P. aeruginosa E. hirae Inoculum Level 7.65 7.69 7.50 7.68 Replicate 1 2 1 2 1 2 1 2 Recovery 2.60 2.52 <2.18 <2.18 <2.18 <2.18 <2.18 <2.18 Log.sub.10 5.05 5.13 >5.51 >5.51 >5.32 >5.32 >5.50 >5.50 Reduction Average Log.sub.10 5.09 >5.51 >5.32 >5.50 Reduction % eradication 99.999 >99.999 >99.999 >99.999

TABLE-US-00005 TABLE 3C tested composition, E3 Organism S. aureus E. coli P. aeruginosa E. hirae Inoculum Level 7.65 7.69 7.50 7.68 Replicate 1 2 1 2 1 2 1 2 Recovery 2.90 2.66 <2.18 <2.18 <2.18 <2.18 <2.18 <2.18 Log.sub.10 4.75 4.99 >5.51 >5.51 >5.32 >5.32 >5.50 >5.50 Reduction Average Log.sub.10 4.87 >5.51 >5.32 >5.50 Reduction % eradication 99.99 >99.999 >99.999 >99.999

TABLE-US-00006 TABLE 3D tested composition, E4 Organism S. aureus E. coli P. aeruginosa E. hirae Inoculum Level 7.69 7.66 7.67 7.68 Replicate 1 2 1 2 1 2 1 2 Recovery <2.18 <2.18 <2.18 <2.18 <2.18 <2.18 <2.18 <2.18 Log.sub.10 >5.51 >5.51 >5.48 >5.48 >5.49 >5.49 >5.50 >5.50 Reduction Average Log.sub.10 >5.51 >5.48 >5.49 >5.50 Reduction % eradication >99.999 >99.999 >99.999 >99.999

TABLE-US-00007 TABLE 3E tested composition: E5 Organism S. aureus E. coli P. aeruginosa E. hirae Inoculum Level 7.67 7.67 7.67 7.66 Replicate 1 2 1 2 1 2 1 2 Recovery <2.18 <2.18 <2.18 <2.18 <2.18 <2.18 <2.18 <2.18 Log.sub.10 >5.49 >5.49 >5.49 >5.49 >5.49 >5.49 >5.48 >5.48 Reduction Average Log.sub.10 >5.49 >5.49 >5.49 >5.48 Reduction % eradication >99.999 >99.999 >99.999 >99.999

[0128] As is seen from the foregoing the compositions according to the invention exhibited excellent antimicrobial efficacy even when the amounts of the cationic surfactant compounds are reduced as compared to the comparative example compositions.

[0129] The compositions of Table 1 all exhibited good foaming characteristics and good hand feel when used as a hand cleaning composition.

[0130] In contrast thereto, the compositions of C1 and C2 did not meet the foregoing acceptance criteria; the C1 and C2 compositions when tested with the foregoing protocol and exhibited an % eradication of one or more of: E. coli, S. aureus, P. aeruginosa, and E. hirae of <90%, and thus did not meet the requirements of the success criteria.

Antimicrobial Efficacy (II)Minimum Inhibitory Concentration:

[0131] A Minimum Inhibitory Concentration evaluation of the example composition E11 was performed using a modification of the Macrodilution Broth Method outlined in CLSI Document M07-A8, Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically, Approved Standard, Eighth Edition. The tested composition was E11, which was challenged versus 27 different microorganism strains, 13 Gram-negative bacterial strains, 13 Gram-Positive bacterial strains, and one yeast species. Each challenge strain was exposed to each of 15 doubling dilutions of the E11 composition which was prepared in sterile nutrient broth. Following incubation, the Minimum Inhibitory Concentration of the test product was determined visually and documented.

[0132] The challenged microorganism strains were as follows:

[0133] Acinetobacter baumannii (ATCC #19606)

[0134] Acinetobacter lwoffii (ATCC #15309)

[0135] Burkholderia cepacia (ATCC #25416)

[0136] Candida albicans (ATCC #10231)

[0137] Corynebacterium jeikeium (ATCC #43734)

[0138] Enterobacter aerogenes (ATCC #13048),

[0139] Enterobacter cloacae cloacae (ATCC #13047),

[0140] Enterococcus faecalis (ATCC #19433)

[0141] Enterococcus faecalis VRE (ATCC #51299)

[0142] Enterococcus faecium (ATCC #19434)

[0143] Escherichia coli (ATCC #11229)

[0144] Escherichia coli (ATCC #25922)

[0145] Haemophilus influenzae (ATCC #19418)

[0146] Klebsiella oxytoca (ATCC #43165)

[0147] Klebsiella pneumoniae ozaenae (ATCC #11296)

[0148] Micrococcus luteus (ATCC #4698)

[0149] Proteus mirabilis (ATCC #7002)

[0150] Pseudomonas aeruginosa (ATCC #9027)

[0151] Serratia marcescens (ATCC #14756)

[0152] Staphylococcus aureus aureus (ATCC #6538)

[0153] Staphylococcus aureus aureus MRSA (ATCC #33592)

[0154] Staphylococcus epidermidis (ATCC #12228)

[0155] Staphylococcus haemolyticus (ATCC #29970)

[0156] Staphylococcus hominis hominis (ATCC #27844)

[0157] Staphylococcus saprophytcus (ATCC #15305)

[0158] Streptococcus pneumonae (ATCC #6303)

[0159] Streptococcus pyogenes (ATCC #19615)

[0160] MRSA=Methicillin-Resistant Staphylococcus aureus

[0161] VRE=Vancomycin-Resistant Enterococcus

[0162] The results of the testing are reported on the following Table 4:

TABLE-US-00008 TABLE 4 Minimum Inhibitory Incoculum Concentration Population (MIC) Challenge Microorganism (per tube; (Expressed As Species ATCC# CFU/mL) Product Dilution) Acinetobacter baumannii 19606 6.0250 10.sup.5 1:1,024 Acinetobacter lwoffii 15309 9.3250 10.sup.5 1:8,192 Burkholderia cepacia 25416 9.550 10.sup.5 1:8 Candida albicans 10231 7.5750 10.sup.5 1:2,048 Corynebacterium jeikeium 43734 2.0250 10.sup.6 1:8,192 Enterobacter aerogenes 13048 8.350 10.sup.5 1:256.sup. Enterobacter cloacae 13047 6.0750 10.sup.5 1:512.sup. cloacae Enterococcus faecalis 19433 9.550 10.sup.5 1:8,192 Enterococcus faecalis VRE 51299 1.450 10.sup.6 1:2,048 Enterococcus faecium 19434 8.850 10.sup.5 1:4,096 Escherichia coli 11229 1.0825 10.sup.6 1:256.sup. Escherichia coli 25922 5.850 10.sup.5 1:512.sup. Haemophilus influenzae 19418 1.2925 10.sup.7 1:2,048 Klebsiella oxytoca 43165 7.20 10.sup.5 1:128.sup. Klebsiella pneumoniae 11296 8.0750 10.sup.5 1:2,048 ozaenae Micrococcus luteus 4698 1.330 10.sup.6 >1:32,768 Proteus mirabilis 7002 1.1125 10.sup.6 1:16 Pseudomonas aeruginosa 9027 7.3750 10.sup.5 1:8 Serratia marcescens 14756 6.20 10.sup.5 1:256.sup. Staphylococcus aureus 6538 1.3950 10.sup.6 1:8,192 aureus Staphylococcus aureus 33592 1.950 10.sup.6 1:2,048 aureus MRSA Staphylococcus epidermidis 12228 8.950 10.sup.5 1:16,384 Staphylococcus 29970 4.2750 10.sup.5 1:16,384 haemolyticus Staphylococcus hominis 27844 3.250 10.sup.5 1:16,384 hominis Staphylococcus 15305 6.6750 10.sup.5 1:16,384 saprophyticus Streptococcus pneumoniae 6303 5.00 10.sup.6 1:2,048 Streptococcus pyogenes 19615 1.2925 10.sup.6 1:4,096 MRSA = Methicillin-Resistant Staphylococcus aureus VRE = Vancomycin-Resistant Enterococcus

[0163] As can be seen from the reported test results of the foregoing table, the composition according to E11 exhibited excellent inhibitory activity, depending upon the microorganism strain evaluated.

Antimicrobial Efficacy (III)In-Vitro Time-Kill:

[0164] The antimicrobial efficacy of example composition E1 of Table 1 (diluted to 90% v/v in distilled water) was evaluated. Neutralization studies of the tested product was performed versus Escherichia coli (ATCC #11229), Staphylococcus aureus (ATCC #6538) and Streptococcus pneumoniae (ATCC #6303) to ensure that the neutralizing solution employed (Butterfield's Phosphate Buffer solution with production neutralizer [BBP++RB]) was effective in reducing the antimicrobial properties of the tested example composition and was non-toxic to each of the challenge microorganism strains used in the test. This neutralization procedure was based on ASTM E-1054-08, Standard Test Methods for Evaluation of Inactivators of Antimicrobial Agents. Under the conditions of this evaluation, the neutralizing solution used (BBP++RB) was demonstrated to be effective in neutralizing the antimicrobial properties of the tested E11 dilution, and was also shown to be non-toxic to all of the challenge microorganism strains used in the test.

[0165] The challenge microorganism strains used in the test were as follows:

[0166] Burkholderia cepacia (ATCC #25416)

[0167] Enterococcus faecalis (ATCC #51299)

[0168] Esherichia coli (ATCC #11229)

[0169] Klebsiella pneumoniae pneumoniae (ATCC #10031)

[0170] Psuedomonas aeruginosa (ATCC #9027)

[0171] Serratia marcescens (ATCC #14756)

[0172] Staphylococcus aureus aureus (ATCC #6538)

[0173] Staphylococcus aureus aureus MRSA (ATCC #33592)

[0174] Staphylococcus epidermidis (ATCC #12228)

[0175] Streptococcus pneumoniae (ATCC #6303)

[0176] Streptococcus pyogenes (ATCC #19615)

[0177] MRSA=Methicillin-Resistant Staphylococcus aureus

[0178] The following Table 5 reports the results of the test, and presents the Initial Population (colony forming units/milliliter=CFU/mL) of the specific challenge microorganism strain tested, the post-exposure population (CFU/mL), and the Log.sub.10 as well as the percent reduction produced by the dilution of the E11 composition 90% v/v) at the times indicated.

TABLE-US-00009 TABLE 5 Post- Inoculum Exposure Challenge Level Exposure Population Log.sub.10 Percent Microorganism (CFU/mL Time (CFU/mL) Reduction Reduction Burkholderia cepacia 2.1550 10.sup.9 30 seconds 4.850 10.sup.8 0.6477 77.4942% (ATCC #25416) 1 minute 2.460 10.sup.8 0.9425 88.5847% Enterococcus faecalis 4.550 10.sup.9 30 seconds <1.00 10.sup.3 6.6580 99.9999% (ATCC #51299) 1 minute <1.00 10.sup.3 6.6580 99.9999% Esherichia coli 1.4650 10.sup.9 30 seconds <1.00 10.sup.3 6.1658 99.9999% (ATCC #11229) 1 minute <1.00 10.sup.3 6.1658 99.9999% Klebsiella pneumoniae 8.750 10.sup.8 30 seconds <1.00 10.sup.3 5.9420 99.9999% pneumoniae 1 minute <1.00 10.sup.3 5.9420 99.9999% (ATCC #10031) Psuedomonas 2.010 10.sup.9 30 seconds <1.00 10.sup.3 6.3032 99.9999% aeruginosa 1 minute <1.00 10.sup.3 6.3032 99.9999% (ATCC #9027) Serratia marcescens 8.10 10.sup.8 30 seconds 2.130 10.sup.7 1.5801 97.3704% (ATCC #14756) 1 minute 1.680 10.sup.6 2.6832 99.7926% Staphylococcus aureus 1.30 10.sup.9 30 seconds 9.00 10.sup.3 5.1597 99.9993% aureus 1 minute <1.00 10.sup.3 6.1139 99.9999% (ATCC #6538) Staphylococcus aureus 1.750 10.sup.9 30 seconds 1.1350 10.sup.5 4.1880 99.9935% aureus MRSA 1 minute 5.50 10.sup.3 5.5026 99.9997% (ATCC #33592) Staphylococcus 7.650 10.sup.8 30 seconds <1.00 10.sup.3 5.8837 99.9999% epidermidis 1 minute <1.00 10.sup.3 5.8837 99.9999% (ATCC #12228) Streptococcus 5.350 10.sup.9 30 seconds <1.00 10.sup.4 5.7284 99.9998% pneumoniae 1 minute <1.00 10.sup.4 5.7284 99.9998% (ATCC #6303) Streptococcus 1.0750 10.sup.9 30 seconds <1.00 10.sup.3 6.0314 99.9999% pyogenes 1 minute <1.00 10.sup.3 6.0314 99.9999% (ATCC #19615)

[0179] As can be seen from the reported test results of the foregoing table, the diluted composition according to E11 exhibited excellent activity, which varied only slightly depending upon the microorganism strain evaluated.

Antimicrobial Efficacy (IV)Handwash Test:

[0180] The antimicrobial efficacy of example composition E11 of Table 1 was evaluated as to its efficacy in the reduction of a challenge microorganism which was evaluated generally in accordance with a test protocol based on 1994 FDA TFM, Health-Care Antiseptic Drug Products; Proposed Rule.

[0181] According to the test protocol, as test subjects were selected fifteen overtly healthy persons at least 18 years of age. All subjects' hands were free from clinically evident dermatoses, injuries to the hands or forearms, hangnails, and/or any other disorders that may have compromised the subject or the study. The test subjects first were subjected to a 7-day pre-test conditioning period followed by a single test day. On the test day, ten product applications of an aliquot of the E11 composition were applied. The test subjects used the E11 composition. over the course of 11 consecutive hand contaminations with an indicator microorganism, Serratia marcescens (ATCC #14756), the first followed by a sample for baseline population, and an additional 10 by product applications 1, 3, 7, and 10. Hand-sampling was performed using the Glove Juice Sampling Procedure.

[0182] The 7 days prior to the test portion of the study constituted the pre-test period. During this time, the test subjects avoided the use of the medicated soaps, lotions, deoterants and shampoos, as well as skin contact with solvents, detergents, acids and bases, or any other skin products known to affect the normal microbial populations of the skin. Subjects were supplied a personal hygiene kit containing nonmedicated soap, shampoo, lotion, and rubber gloves to be worn when contact with antimicrobials, solvents, detergents, acids, or bases could not be avoided. Subjects were instructed to use the contents of this kit exclusively during their participation in the study. Subjects also avoided using UV tanning beds or sunbathing, and swimming or bathing in biocide-treated pools or hot tubs.

[0183] In preparation for the tests performed on the test day, an inoculum of the indicator microorganism, Serratia marcescens (ATCC #14756) was prepared for use to challenge the efficacy of the E11 composition.

[0184] A stock culture of Serratia marcescens (ATCC #14756) was prepared by aseptically transferring contents of a lyophilized vial to approximately 5.0 mL of sterile Tryptic Soy Broth (TSB), which was then incubated at 25 C. 2 C. for 20 hours. 2-L flasks containing approximately 1,000 mL TSB were inoculated with 1.0 mL of the 20-hour broth cultures, and incubated fro 21-26 hours at 25 C.2 C. Prior to any withdrawal of culture, whether for hand contamination or for numbers assay, the suspensions were stirred or swirled. The suspensions were assayed for number of organisms at the beginning and end of the use periods. Suspensions were not used for more than 8 hours.

[0185] On the test day, prior to being admitted into testing, subjects were questioned regarding their adherence to the Protocol requirements. Subjects clipped their fingernails to a free edge of 1 mm, if they had not already done so. All jewelry was removed from the hands and arms prior to washing. Using tap water, subjects performed a practice hand-inoculation procedure. A 5.0-mL aliquot of tap water was transferred into each subject's cupped hands in three volumes of approximately 1.5 mL, 1.5 mL, and 2 mL. A 30-second handwash using nonmedicated soap and a 30-second rinse were performed to remove dirt and oil from hands. The temperature of the water used for this and subsequent wash procedures was controlled at 40 C.2 C.

[0186] Thereafter, a 5.0-mL aliquot of a suspension containing approximately 1.010.sup.9 CFU/mL of Serratia marcescens (ATCC #14756) was transferred into each subjects cupped hands in three volumes of approximately 1.5 mL, 1.5 mL, and 2 mL. The inoculum distributed evenly over both hands, not reaching above the wrists, via gentle continuous massage for 45 seconds. After a timed 2-minute air-dry, and within 5 minutes after contamination for baseline, and/or after product applications 1, 3, 7, and 10, powder-free sterile latex gloves were placed on subjects' hands, and 75 mL of Stripping Suspending Fluid (SSF) were instilled into each of the gloves. The wrists were secured, and technicians massaged the hands through the gloves in a standardized manner for 60 seconds. Within 1 minute of completing the post-application massages, a 5-mL aliquot of the glove juice was removed from each of the gloves and diluted in 5.0 mL Butterfield's Phosphate Buffer Solution with product neutralizers (BBP++) (dilution 10). The 10 dilutions were then serially diluted in BBP++. Duplicate spiral plates were prepared from specified dilutions on Tryptic Soy Agar with product neutralizers (TSA+) and incubated at 25 C. 2 C. for approximately 40 hours, when sufficient growth was observed. Colonies were counted and data recorded using the computerized QCOUNT plate-counting system.

[0187] The first contamination cycle provide the baseline recovery populations. It was followed with a 30-second handwash using nonmedicated soap and a 30-second rinse.

[0188] A 5.0-mL aliquot of the microbial inoculum was again evenly distributed over both hands, not reaching above the wrists, via gentle continuous massage for 45 seconds. After a timed 2-minute air-dry, the subjects applied 5 mL of the E11 composition which was e dispensed into the test subjects' cupped hands. Thereafter the test subjects rubbed hands together, lathering hands and forearms to just above the wrist for 30 seconds. Subjects rinsed for 30 seconds. The hands were gloved wet for sampling. If product application was not followed by a sample, the test subjects' dried their hands with a paper towel.

[0189] Each test subject completed this contamination/product application a total of 10 consecutive times, with a minimum of 5 minutes between contamination/product applications. The test subjects' hands were sampled for residual Serratia marcescens (ATCC #14756) after contamination/product application cycles 1, 3, 7, and 10. Sampling was performed after Applications 1, 3, 7, and 10.

All sampling was performed using the aforementioned protocol wherein powder-free sterile latex gloves were placed on subjects' hands, SSF was instilled, the wrists secured, the hands massaged, and within 1 minute thereafter a 5-mL aliquot removed, neutralized and serially diluted.

[0190] The performance of the E11 composition was evaluated post-application based upon mean log.sub.10 reductions from baseline. Log.sub.10 reductions were calculated by subtracting the log.sub.10 number of bacteria recovered at the specified sampling times following product application (applications 1, 3, 7, and 10) from the log.sub.10 number of bacteria recovered from the baseline-sampling. The critical index was a mean log.sub.10 reduction in bacteria of 2 log.sub.10 after the first application of the E11 composition. The Minitab computer package was used for all statistical calculations. Statistical calculations of mean, standard deviation, and 95% confidence intervals were performed on the log.sub.10 microbial recoveries in baseline and post-product application samples, and the reductions from the baseline. The Log.sub.10 microbial recoveries from each subject's hands and reductions from baseline populations are presented in the following Table 6 which provides the mean Log.sub.10 microbial recoveries of Serratia marcescens (ATCC #14756) and the mean Log.sub.10 reductions from the baseline following the use of the E11 composition, reported for each test subject, and each hand.

TABLE-US-00010 TABLE 6 Post- Post- Post- Post- Post- Post- Post- Post- Wash 1 Wash 3 Wash 7 Wash 10 Baseline Wash 1 Wash 3 Wash 7 Wash 10 Log.sub.10 Log.sub.10 Log.sub.10 Log.sub.10 Log.sub.10 Log.sub.10 Log.sub.10 Log.sub.10 Log.sub.10 Reduction Reduction Reduction Reduction Microbial Microbial Microbial Microbial Microbial from from from from Subject Hand Recovery Recovery Recovery Recovery Recovery Baseline Baseline Baseline Baseline 2 Left 9.00 6.74 6.28 5.81 5.74 2.26 2.72 3.19 3.26 Right 8.89 6.50 5.97 5.45 5.92 2.39 2.91 3.44 2.97 4 Left 9.15 6.33 6.07 5.44 5.33 2.82 3.08 3.72 3.82 Right 9.28 5.98 5.80 5.36 5.54 3.30 3.48 3.92 3.74 5 Left 9.27 6.86 6.37 5.94 5.79 2.41 2.90 3.33 3.48 Right 9.31 6.45 6.21 5.80 5.51 2.86 3.10 3.51 3.81 6 Left 8.96 6.62 6.55 5.75 5.84 2.34 2.41 3.21 3.12 Right 8.90 6.49 6.30 6.00 6.09 2.42 4.60 2.90 2.82 10 Left 9.34 6.05 5.62 5.10 5.18 3.30 3.72 4.24 4.16 Right 9.24 6.38 5.41 4.91 4.82 2.85 3.83 4.33 4.42 11 Left 8.83 6.18 6.02 5.80 5.28 2.64 2.81 3.03 3.54 Right 8.81 6.55 6.35 5.63 5.46 2.26 2.46 3.18 3.36 13 Left 9.36 6.80 6.48 6.16 6.05 2.56 2.88 3.20 3.31 Right 9.38 6.42 6.33 6.10 5.87 2.96 3.05 3.29 3.51 17 Left 9.23 6.66 6.60 6.13 6.17 2.58 2.64 3.10 3.06 Right 9.24 6.21 6.43 5.93 5.87 3.03 2.81 3.31 3.37 20 Left 9.24 6.66 6.25 6.01 5.99 2.58 2.99 3.22 3.25 Right 9.24 6.33 6.02 5.65 5.62 2.91 3.22 3.59 3.62 21 Left 9.17 6.11 * 5.84 5.72 3.06 * 3.33 3.45 Right 9.04 6.13 * 5.68 5.65 2.91 * 3.36 3.39 23 Left 8.98 6.12 6.06 5.64 5.57 2.86 2.91 3.34 3.40 Right 9.01 6.35 5.92 5.46 5.34 2.66 3.09 3.55 3.67 26 Left 9.32 6.53 6.03 ** 5.46 2.79 3.29 ** 3.86 Right 9.20 6.77 6.36 6.10 5.59 2.43 2.84 3.10 3.61 27 Left 9.33 6.37 5.66 5.53 5.47 2.97 3.67 3.80 3.87 Right 9.34 6.17 5.81 5.39 5.41 3.17 3.53 3.95 3.93 28 Left 8.99 6.18 5.85 5.53 5.33 2.80 3.13 3.46 3.66 Right 9.02 6.00 5.60 5.31 4.90 3.02 3.42 3.72 4.12 29 Left 9.06 5.94 6.01 5.18 5.78 3.12 3.05 3.88 3.29 Right 9.07 6.10 5.56 5.31 5.40 2.97 3.52 3.76 3.68 * Data not used in analysis due to deviation from protocol ** Not used in analysis due to apparent error

[0191] As can be understood with reference to the results reported on Table 6, the E11 composition produced a mean log.sub.10 reduction of Serratia marcescens (ATCC #14756) of 2.77 after Application 1, indicating good efficacy against this indicator microorganism on hands of human test subjects.

Antimicrobial Efficacy (V)EN1276 Test:

[0192] The antimicrobial efficacy of example composition E9 at three different dilutions (10% v/v, 50% v/v and 80% v/v, in distilled water) was evaluated in accordance with the EN1276 (:2012) Standard Suspension Test for bactericidal activity against Staphylococcus aureus (ATCC #6538), Pseudomonas aeruginosa (ATCC #15442), Escherichia coli (ATCC #10536) and Enterococcus hirae (ATCC #10541). Samples of each test composition were placed in test tube with 3% BSA (dirty conditions) and challenge organism to achieve the indicated dilutions for a contact time of 1 minute, followed by neutralization, serial dilutions, plating and incubation. The test was performed at room temperature (approx. 20 C.+/0.6 C.). The results of the tested compositions are reported on the following Table 7.

TABLE-US-00011 TABLE 7 validation test dilution- microbial test procedure at product bacterial Test neutralizer neutralization suspension concentration V/V test strain suspension conditions control control in the test 10% 50% 80% Pseudomonas aeruginosa Vc: 115; 139 Vc: 119; 126 Vc: 108;120 Vc: 85; 103 10.sup.6: >330; >330 Vc >330; >330 0; 0 0; 0 PCM 2563 = Nv.sub.0: 1.3 .Math. 10.sup.2 A: 1.2 .Math. 10.sup.2 B: 1.1 .Math. 10.sup.2 C: 9.4 .Math. 10.sup.1 10.sup.7: 42; 55 Na >3300 <140 <140 ATCC 15442 N: 4.8 .Math. 10.sup.8 Ig Na >3.52 <2.15 <2.15 N.sub.0 4.8 .Math. 10.sup.7 R <4.16 >5.53 >5.53 IgN.sub.0 = 7.68 Staphylococcus aureus Vc: 103; 110 Vc: 127; 144 Vc: 98; 114 Vc; 89; 96 10.sup.6: >330; >330 Vc 8; 13 5; 6 15; 21 ATCC 6538 Nv.sub.0: 1.1 .Math. 10.sup.2 A: 1.4 .Math. 10.sup.2 B: 1.1 .Math. 10.sup.2 C: 9.2 .Math. 10.sup.1 10.sup.7: 39; 46 Na <140 <140 180 N: 4.2 .Math. 10.sup.8 Ig Na <2.15 <2.15 2.26 N.sub.0.Math.4.2 .Math. 10.sup.7 R >5.47 >5.47 5.36 IgN.sub.0 = 7.62 Escherichia coli Vc: 65; 78 Vc: 68; 73 Vc: 57; 70 Vc: 47; 66 10.sup.6: 254; 271 Vc 0; 0 0; 0 0; 0 PCM 1144 = Nv.sub.0: 7.2 .Math. 10.sup.1 A: 7.0 .Math. 10.sup.1 B: 6.4 .Math. 10.sup.1 C: 5.6 .Math. 10.sup.1 10.sup.7: 30; 36 Na <140 <140 <140 ATCC 10536 N: 2.7 .Math. 10.sup.8 Ig Na <2.15 <2.15 <2.15 N.sub.0 2.7 .Math. 10.sup.7 R >5.28 >5.28 >5.28 IgN.sub.0 = 7.43 Enterococcus hirae Vc: 42; 56 Vc. 38; 46 Vc: 48; 53 Vc: 62; 68 10.sup.6: 174; 198 Vc 0; 0 0; 0 0; 0 PCM 2559 = Nv.sub.0: 4.9 .Math. 10.sup.1 A: 4.2 .Math. 10.sup.1 B: 5.0 .Math. 10.sup.1 C: 6.5 .Math. 10.sup.1 10.sup.7: 12; 19 Na <140 <140 <140 ATCC 10541 N: 1.9 .Math. 10.sup.8 Ig Na <2.15 <2.15 <2.15 N.sub.0 1.9 .Math. 10.sup.7 R >5.13 >5.13 >5.13 IgN.sub.0 = 7.28 Vcviable count Ncfu/ml in the bacterial test suspension, N.sub.0 = N/10 Nvcfu/ml in the bacterial suspension in validation procedure, Nv.sub.0 = Nv/10 Rreduction factor of viable counts Nacfu/ml in the test mixture Acfu/ml in the experimental conditions control test Bcfu/ml in neutralizer control test Ccfu/ml in the dilation - neutralization method control test Validation criteria verification: 1.5 .Math. 10.sup.8 < N < 5.0 .Math. 10.sup.8 - met 3.0 .Math. 10.sup.2 < Nv < 1.6 .Math. 10.sup.3 - met A, B, C > 0.5 Nv.sub.0 - met

Antimicrobial Efficacy (VI)EN1276 Test:

[0193] The antimicrobial efficacy of example composition E10 at three different dilutions (10% v/v, 50% v/v and 80% v/v, in distilled water) was evaluated in accordance with the EN1276 Standard Suspension Test for bactericidal activity against Salmonella enterica (ATCC #13311), Staphylococcus aureus (ATCC #33592), Staphylococcus aureus (ATCC #6538), Klebsiella pneumoniae (ATCC BAA 2146), Pseudomonas aeruginosa (ATCC #15442), Escherichia coli (ATCC #10536) and Enterococcus hirae (ATCC #10541). Samples of each test composition were placed in test tube with 3% BSA (dirty conditions) and challenge organism to achieve the indicated dilutions for a contact time of 1 minute, followed by neutralization, serial dilutions, plating and incubation. The test was performed at room temperature (approx. 20 C.+/0.6 C.). The results of the tested compositions are reported on the following Tables 8 and 9.

TABLE-US-00012 TABLE 8 validation test dilution- microbial test procedure at product bacterial Test neutralizer neutralization suspension concentration V/V test strain suspension conditions control control in the test 10% 50% 80% Pseudomonas aeruginosa Vc: 115; 139 Vc: 119; 126 Vc: 108; 120 Vc: 124; 142 10.sup.6: >330; >330 Vc >330; >330 0; 0 0; 0 PCM 2563 = Nv.sub.0: 1.3 .Math. 10.sup.2 A: 1.2 .Math. 10.sup.2 B: 1.1 .Math. 10.sup.2 C: 1.3 .Math. 10.sup.2 10.sup.7: 42; 55 Na >3300 <140 <140 ATCC 15442 N: 4.8 .Math. 10.sup.8 Ig Na >3.52 <2.15 <2.15 N.sub.0 4.8 .Math. 10.sup.7 R <4.16 >5.53 >5.53 IgN.sub.0 = 7.68 Staphylococcus aureus Vc: 103; 110 Vc: 127; 144 Vc: 98; 114 Vc: 86; 103 10.sup.6: >330; >330 Vc 7; 11 17; 24 14; 20 ATCC 6538 Nv.sub.0: 1.1 .Math. 10.sup.2 A: 1.4 .Math. 10.sup.2 B: 1.1 .Math. 10.sup.2 C: 9.4 .Math. 10.sup.1 10.sup.7: 39; 46 Na <140 205 170 N: 4.2 .Math. 10.sup.8 Ig Na <2.15 2.31 2.23 N.sub.0 4.2 .Math. 10.sup.7 R >5.47 >5.31 5.39 IgN.sub.0 = 7.62 Escherichia coli Vc: 65; 78 Vc: 68; 73 Vc: 57; 70 Vc: 81; 86 10.sup.6: 254; 271 Vc 0; 0 0; 0 0; 0 PCM 1144 = Nv.sub.0: 7.2 .Math. 10.sup.1 A: 7.0 .Math. 10.sup.1 B: 6.4 .Math. 10.sup.1 C: 8.4 .Math. 10.sup.1 10.sup.7: 30; 36 Na <140 <140 <140 ATCC 10536 N: 2.7 .Math. 10.sup.8 Ig Na <2.15 <2.15 <2.15 N.sub.0 2.7 .Math. 10.sup.7 R >5.28 >5.28 >5.28 IgN.sub.0 = 7.43 Enterococcus hirae Vc: 42; 56 Vc: 38; 46 Vc: 48; 53 Vc: 31; 44 10.sup.6: 174; 198 Vc 5; 12 0; 0 1; 5 PCM 2559 = Nv.sub.0: 4.9 .Math. 10.sup.1 A: 4.2 .Math. 10.sup.1 B: 5.0 .Math. 10.sup.1 C: 3.8 .Math. 10.sup.1 10.sup.7: 12; 19 Na <140 <140 <140 ATCC 10541 N: 1.9 .Math. 10.sup.8 Ig Na <2.15 <2.15 <2.15 N.sub.0 1.9 .Math. 10.sup.7 R >5.13 >5.13 >5.13 IgN.sub.0 = 7.28 Vcviable count Ncfu/ml in the bacterial test suspension, N.sub.0 = N/10 Nvcfu/ml in the bacterial suspension in validation procedure, Nv.sub.0 = Nv/10 Rreduction factor of viable counts Nacfu/ml in the test mixture Acfu/ml in the experimental conditions control test Bcfu/ml in neutralizer control test Ccfu/ml in the dilation - neutralization method control test Validation criteria verification: 1.5 .Math. 10.sup.8 < N < 5.0 .Math. 10.sup.8 - met 3.0 .Math. 10.sup.2 < Nv < 1.6 .Math. 10.sup.3 - met A, B, C > 0.5 Nv.sub.0 - met

TABLE-US-00013 TABLE 9 validation test dilution- microbial test procedure at product bacterial Test neutralizer neutralization suspension concentration V/V test strain suspension conditions control control in the test 10% 50% 80% Staphylococcus aureus Vc: 127; 145 Vc: 134; 158 Vc: 113; 120 Vc: 125; 132 10.sup.6: >330; >330 Vc 0; 0 0; 0 0; 2 ATCC 33592 Nv.sub.0: 1.4 .Math. 10.sup.2 A: 1.5 .Math. 10.sup.2 B: 1.2 .Math. 10.sup.2 C: 1.3 .Math. 10.sup.2 10.sup.7: 42; 54 Na <140 <140 <140 N: 4.8 .Math. 10.sup.8 Ig Na <2.15 <2.15 <2.15 N.sub.0 4.8 .Math. 10.sup.7 R >5.53 >5.53 >5.53 IgN.sub.0 = 7.68 Salmonella enterica sb. Vc: 102; 115 Vc: 84; 108 Vc: 121; 127 Vc: 98; 114 10.sup.6: >330; >330 Vc 0; 0 0; 0 0; 0 enterica Nv.sub.0: 1.1 .Math. 10.sup.2 A: 9.6 .Math. 10.sup.1 B: 1.2 .Math. 10.sup.2 C: 1.1 .Math. 10.sup.2 10.sup.7: 39; 45 Na <140 <140 <140 PCM 2565 = N: 4.2 .Math. 10.sup.8 Ig Na <2.15 <2.15 <2.15 ATCC 13311 N.sub.0 4.2 .Math. 10.sup.7 R >5.47 >5.47 >5.47 IgN.sub.0 = 7.62 Klebsiella pneumoniae Vc: 59; 74 Vc: 54; 68 Vc: 71; 79 Vc: 61; 64 10.sup.6: 227; 251 Vc >330; >330 0; 0 0; 0 ATCC BAA - 2146 Nv.sub.0: 6.6 .Math. 10.sup.1 A: 6.1 .Math. 10.sup.1 B: 7.5 .Math. 10.sup.1 C: 6.2 .Math. 10.sup.1 10.sup.7: 25; 28 Na >3300 <140 <140 N: 2.4 .Math. 10.sup.8 Ig Na >3.52 <2.15 <2.15 N.sub.0 2.4 .Math. 10.sup.7 R <3.86 >5.23 >5.23 IgN.sub.0 = 7.38 Vcviable count Ncfu/ml in the bacterial test suspension, N.sub.0 = N/10 Nvcfu/ml in the bacterial suspension in validation procedure, Nv.sub.0 = Nv/10 Rreduction factor of viable counts Nacfu/ml in the test mixture Acfu/ml in the experimental conditions control test Bcfu/ml in neutralizer control test Ccfu/ml in the dilation - neutralization method control test Validation criteria verification: 1.5 .Math. 10.sup.8 < N < 5.0 .Math. 10.sup.8 - met 3.0 .Math. 10.sup.2 < Nv < 1.6 .Math. 10.sup.3 - met A, B, C > 0.5 Nv.sub.0 - met

Antimicrobial Efficacy (VII)EN1276 Test:

[0194] The antimicrobial efficacy of example composition E11 at three different dilutions (10% v/v, 50% v/v and 80% v/v, in distilled water) was evaluated in accordance with the EN1276 Standard Suspension Test for bactericidal activity against Salmonella enterica (ATCC #13311), Staphylococcus aureus (ATCC #33592), Staphylococcus aureus (ATCC #6538), Klebsiella pneumoniae (ATCC BAA 2146), Pseudomonas aeruginosa (ATCC #15442), Escherichia coli (ATCC #10536) and Enterococcus hirae (ATCC #10541). Samples of each test composition were placed in test tube with 3% BSA (dirty conditions) and challenge organism to achieve the indicated dilutions for a contact time of 1 minute, followed by neutralization, serial dilutions, plating and incubation. The test was performed at room temperature (approx. 20 C.+/0.6 C.). The results of the tested compositions are reported on the following Tables 10 and 11.

TABLE-US-00014 TABLE 10 validation test dilution- microbial test procedure at product bacterial Test neutralizer neutralization suspension concentration V/V test strain suspension conditions control control in the test 10% 50% 80% Pseudomonas aeruginosa Vc: 115; 139 Vc: 119; 126 Vc: 108; 120 Vc: 104; 128 10.sup.6: >330; >330 Vc >330; >330 0; 0 0; 0 PCM 2563 = Nv.sub.0: 1.3 .Math. 10.sup.2 A: 1.2 .Math. 10.sup.2 B: 1.1 .Math. 10.sup.2 C: 1.2 .Math. 10.sup.2 10.sup.7: 42; 55 Na >3300 <140 <140 ATCC 15442 N: 4.8 .Math. 10.sup.8 Ig Na >3.52 <2.15 <2.15 N.sub.0 4.8 .Math. 10.sup.7 R <4.16 >5.53 >5.53 IgN.sub.0 = 7.68 Staphylococcus aureus Vc: 103; 110 Vc: 127; 144 Vc: 98; 144 Vc: 112; 125 10.sup.6: >330; >330 Vc 0; 4 13; 17 6; 9 ATCC 6538 Nv.sub.0: 1.1 .Math. 10.sup.3 A: 1.4 .Math. 10.sup.2 B: 1.1 .Math. 10.sup.3 C: 1.2 .Math. 10.sup.2 10.sup.7: 39; 46 Na <140 150 <140 N: 4.2 .Math. 10.sup.8 Ig Na <2.15 2.18 <2.15 N.sub.0 4.2 .Math. 10.sup.7 R >5.47 5.44 >5.47 IgN.sub.0 = 7.62 Escherichia coli Vc: 65; 78 Vc: 68; 73 Vc: 57; 70 Vc: 49; 54 10.sup.6: 254; 271 Vc 0; 0 0; 1 0; 0 PCM 1144 = Nv.sub.0: 7.2 .Math. 10.sup.1 A: 7.0 .Math. 10.sup.1 B: 6.4 .Math. 10.sup.1 C: 5.2 .Math. 10.sup.1 10.sup.7: 30; 36 Na <140 <140 <140 ATCC 10536 N: 2.7 .Math. 10.sup.8 Ig Na <2.15 <2.15 <2.15 N.sub.0 2.7 .Math. 10.sup.7 R >5.28 >5.28 >5.28 IgN.sub.0 = 7.43 Enterococcus hirae Vc: 42; 56 Vc: 38; 46 Vc: 48, 53 Vc: 35; 51 10.sup.6: 174; 198 Vc 0; 0 0; 0 0; 0 PCM 2559 = Nv.sub.0: 4.9 .Math. 10.sup.1 A: 4.2 .Math. 10.sup.1 B: 5.0 .Math. 10.sup.1 C: 4.3 .Math. 10.sup.1 10.sup.7: 12; 19 Na <140 <140 <140 ATCC 10541 N: 1.9 .Math. 10.sup.8 Ig Na <2.15 <2.15 <2.15 N.sub.0 1.9 .Math. 10.sup.7 R >5.13 >5.13 >5.13 IgN.sub.0 = 7.28 Vcviable count Ncfu/ml in the bacterial test suspension, N.sub.0 = N/10 Nvcfu/ml in the bacterial suspension in validation procedure, Nv.sub.0 = Nv/10 Rreduction factor of viable counts Nacfu/ml in the test mixture Acfu/ml in the experimental conditions control test Bcfu/ml in neutralizer control test Ccfu/ml in the dilation - neutralization method control test Validation criteria verification: 1.5 .Math. 10.sup.8 < N < 5.0 .Math. 10.sup.8 - met 3.0 .Math. 10.sup.2 < Nv < 1.6 .Math. 10.sup.3 - met A, B, C > 0.5 Nv.sub.0 - met

TABLE-US-00015 TABLE 11 validation test dilution- microbial test procedure at product bacterial Test neutralizer neutralization suspension concentration V/V test strain suspension conditions control control in the test 10% 50% 80% Staphylococcus aureus Vc: 127; 145 Vc: 134; 158 Vc: 113; 120 Vc: 106; 109 10.sup.6: >330; >330 Vc 0; 0 3.5 0; 0 ATCC 33592 Nv.sub.0: 1.4 .Math. 10.sup.2 A: 1.5 .Math. 10.sup.3 B: 1.2 .Math. 10.sup.2 C: 1.1 .Math. 10.sup.2 10.sup.7: 42; 54 Na <140 <140 <140 N: 4.8 .Math. 10.sup.8 Ig Na <2.15 <2.15 <2.15 N.sub.0 4.8 .Math. 10.sup.7 R >5.53 >5.53 >5.53 IgN.sub.0 = 7.68 Salmonella enterica sb. Vc: 102; 115 Vc: 84; 108 Vc: 121; 127 Vc: 117; 135 10.sup.6: >330; >330 Vc 0; 0 0; 0 0; 0 enterica Nv.sub.0: 1.1 .Math. 10.sup.2 A: 9.6 .Math. 10.sup.1 B: 1.2 .Math. 10.sup.2 C: 1.3 .Math. 10.sup.2 10.sup.7: 39; 45 Na <140 <140 <140 PCM 2565 = N: 4.2 .Math. 10.sup.8 Ig Na <2.15 <2.15 <2.15 ATCC 13311 N.sub.0 4.2 .Math. 10.sup.7 R >5.47 >5.47 >5.47 IgN.sub.0 = 7.62 Klebsiella pneumoniae Vc: 59; 74 Vc: 54; 68 Vc: 71; 79 Vc: 67; 80 10.sup.6: 227; 251 Vc >330; >330 0; 0 0; 0 ATCC BAA - 2146 Nv.sub.0: 6.6 .Math. 10.sup.1 A: 6.1 .Math. 10.sup.1 B: 7.5 .Math. 10.sup.1 C: 7.4 .Math. 10.sup.1 10.sup.7: 25; 28 Na >3300 <140 <140 N: 2.4 .Math. 10.sup.8 Ig Na >3.52 <2.15 <2.15 N.sub.0 2.4 .Math. 10.sup.7 R <3.86 >5.23 >5.23 IgN.sub.0 = 7.38 Vcviable count Ncfu/ml in the bacterial test suspension, N.sub.0 = N/10 Nvcfu/ml in the bacterial suspension in validation procedure, Nv.sub.0 = Nv/10 Rreduction factor of viable counts Nacfu/ml in the test mixture Acfu/ml in the experimental conditions control test Bcfu/ml in neutralizer control test Ccfu/ml in the dilation - neutralization method control test Validation criteria verification: 1.5 .Math. 10.sup.8 < N < 5.0 .Math. 10.sup.8 - met 3.0 .Math. 10.sup.2 < Nv < 1.6 .Math. 10.sup.3 - met A, B, C > 0.5 Nv.sub.0 - met

Antimicrobial Efficacy (VIII)EN1276 Test:

[0195] The antimicrobial efficacy of example composition E12 at three different dilutions (10% v/v, 50% v/v and 80% v/v, in distilled water) was evaluated in accordance with the EN1276 Standard Suspension Test for bactericidal activity against Salmonella enterica (ATCC #13311), Staphylococcus aureus (ATCC #33592), Staphylococcus aureus (ATCC #6538), Klebsiella pneumoniae (ATCC BAA 2146), Pseudomonas aeruginosa (ATCC #15442), Escherichia coli (ATCC #10536) and Enterococcus hirae (ATCC #10541). Samples of each test composition were placed in test tube with 3% BSA (dirty conditions) and challenge organism to achieve the indicated dilutions for a contact time of 1 minute, followed by neutralization, serial dilutions, plating and incubation. The test was performed at room temperature (approx. 20 C.+/0.6 C.). The results of the tested compositions are reported on the following Tables 12 and 13.

TABLE-US-00016 TABLE 12 validation test dilution- microbial test procedure at product bacterial Test neutralizer neutralization suspension concentration V/V test strain suspension conditions control control in the test 10% 50% 80% Pseudomonas aeruginosa Vc: 115; 139 Vc: 119; 126 Vc: 108; 120 Vc: 93; 98 10.sup.6: >330; >330 Vc >330; >330 0; 0 0; 0 PCM 2563 = Nv.sub.0: 1.3 .Math. 10.sup.2 A: 1.2 .Math. 10.sup.2 B: 1.1 .Math. 10.sup.2 C: 9.6 .Math. 10.sup.1 10.sup.7: 42; 55 Na >3300 <140 <140 ATCC 15442 N: 4.8 .Math. 10.sup.8 Ig Na >3.52 <2.15 <2.15 N.sub.0 4.8 .Math. 10.sup.7 R <4.16 >5.53 >5.53 IgN.sub.0 = 7.68 Staphylococcus aureus Vc: 103; 110 Vc: 127; 144 Vc: 98; 114 Vc: 71; 84 10.sup.6: >330; >330 Vc 1; 6 32; 35 23; 31 ATCC 6538 Nv.sub.0: 1.1 .Math. 10.sup.2 A: 1.4 .Math. 10.sup.2 B: 1.1 .Math. 10.sup.2 C: 7.8 .Math. 10.sup.1 10.sup.7: 39; 46 Na <140 335 270 N: 4.2 .Math. 10.sup.8 Ig Na <2.15 2.53 2.43 N.sub.0 4.2 .Math. 10.sup.7 R >5.47 5.09 5.19 IgN.sub.0 = 7.62 Escherichia coli Vc: 65; 78 Vc: 68; 73 Vc: 57; 70 Vc: 53; 61 10.sup.6: 254; 271 Vc 0; 0 0; 0 0; 0 PCM 1144 = Nv.sub.0: 7.2 .Math. 10.sup.1 A: 7.0 .Math. 10.sup.1 B: 6.4 .Math. 10.sup.1 C: 5.7 .Math. 10.sup.1 10.sup.7: 30; 36 Na <140 <140 <140 ATCC 10536 N: 2.7 .Math. 10.sup.8 Ig Na <2.15 <2.15 <2.15 N.sub.0 2.7 .Math. 10.sup.7 R >5.28 >5.28 >5.28 IgN.sub.0 = 7.43 Enterococcus hirae Vc: 42; 56 Vc: 38; 46 Vc: 48; 53 Vc: 35; 47 10.sup.6: 174; 198 Vc 0; 0 0; 0 0; 0 PCM 2559 = Nv.sub.0: 4.9 .Math. 10.sup.1 A: 4.2 .Math. 10.sup.1 B: 5.0 .Math. 10.sup.1 C: 4.1 .Math. 10.sup.1 10.sup.7: 12; 19 Na <140 <140 <140 ATCC 10541 N: 1.9 .Math. 10.sup.8 Ig Na <2.15 <2.15 <2.15 N.sub.0 1.9 .Math. 10.sup.7 R >5.13 >5.13 >5.13 IgN.sub.0 = 7.28 Vcviable count Ncfu/ml in the bacterial test suspension, N.sub.0 = N/10 Nvcfu/ml in the bacterial suspension in validation procedure, Nv.sub.0 = Nv/10 Rreduction factor of viable counts Nacfu/ml in the test mixture Acfu/ml in the experimental conditions control test Bcfu/ml in neutralizer control test Ccfu/ml in the dilation - neutralization method control test Validation criteria verification: 1.5 .Math. 10.sup.8 < N < 5.0 .Math. 10.sup.8 - met 3.0 .Math. 10.sup.2 < Nv < 1.6 .Math. 10.sup.3 - met A, B, C > 0.5 Nv.sub.0 - met

TABLE-US-00017 TABLE 13 validation test dilution- microbial test procedure at product bacterial Test neutralizer neutralization suspension concentration V/V test strain suspension conditions control control in the test 10% 50% 80% Staphylococcus aureus Vc: 127; 145 Vc: 134; 158 Vc: 113; 120 Vc: 96; 101 10.sup.6: >330; >330 Vc 4; 6 0; 0 1; 1 ATCC 33592 Nv.sub.0: 1.4 .Math. 10.sup.2 A: 1.5 .Math. 10.sup.2 B: 1.2 .Math. 10.sup.2 C: 9.8 .Math. 10.sup.1 10.sup.7: 42; 54 Na <140 <140 <140 N: 4.8 .Math. 10.sup.8 Ig Na <2.15 <2.15 <2.15 N.sub.0 4.8 .Math. 10.sup.7 R >5.53 >5.53 >5.53 IgN.sub.0 = 7.68 10.sup.6: >330; >330 Vc 0; 0 0; 0 0; 0 Salmonella enterica sb. Vc: 102; 115 Vc: 84; 108 Vc: 121; 127 Vc: 81; 93 10.sup.7: 39; 45 Na <140 <140 <140 enterica Nv.sub.0: 1.1 .Math. 10.sup.2 A: 9.6 .Math. 10.sup.1 B: 1.2 .Math. 10.sup.2 C: 8.7 .Math. 10.sup.1 N: 4.2 .Math. 10.sup.8 Ig Na <2.15 <2.15 <2.15 PCM 2565 = N.sub.0 4.2 .Math. 10.sup.7 R >5.47 >5.47 >5.47 ATCC 13311 IgN.sub.0 = 7.62 Klebsiella pneumoniae Vc: 59; 74 Vc: 54; 68 Vc: 71; 79 Vc: 52; 55 10.sup.6: 227; 251 Vc >330; >330 0; 0 0; 0 ATCC BAA - 2146 Nv.sub.0: 6.6 .Math. 10.sup.1 A: 6.1 .Math. 10.sup.1 B: 7.5 .Math. 10.sup.1 C: 5.4 .Math. 10.sup.1 10.sup.7: 25; 28 Na >3300 <140 <140 N: 2.4 .Math. 10.sup.8 Ig Na >3.52 <2.15 <2.15 N.sub.0 2.4 .Math. 10.sup.7 R <3.86 >5.23 >5.23 IgN.sub.0 = 7.38 Vcviable count Ncfu/ml in the bacterial test suspension, N.sub.0 = N/10 Nvcfu/ml in the bacterial suspension in validation procedure, Nv.sub.0 = Nv/10 Rreduction factor of viable counts Nacfu/ml in the test mixture Acfu/ml in the experimental conditions control test Bcfu/ml in neutralizer control test Ccfu/ml in the dilation - neutralization method control test Validation criteria verification: 1.5 .Math. 10.sup.8 < N < 5.0 .Math. 10.sup.8 - met 3.0 .Math. 10.sup.2 < Nv < 1.6 .Math. 10.sup.3 - met A, B, C > 0.5 Nv.sub.0 - met

Antimicrobial Efficacy (IX)Handwash Test:

[0196] The antimicrobial efficacy of example composition E11 of Table 1 was evaluated as to its efficacy in the reduction of a challenge microorganism in accordance with the test protocols of EN 1499 (:2000) Hygienic Handwash. The test utilized 15 human subjects each having a left and right hand. The antimicrobial performance of the E11 composition was compared to that of a reference soft soap (having a density of 200 g/litre) as outlined in EN 1449. The E11 compositions were tested without further dilution and evaluated for antimicrobial performance against the challenge microorganism Escherichia coli (PCM 2560=NCTC 10538). The test was performed at room temperature and all materials used in the test were also at room temperature (20 C.+/0.6 C.). An aliquot (3 ml) of undiluted E11 was applied to the subject's hands, and the handwash protocols of EN 1449Appendix A handwashing continued for 30 seconds, after which the subject rinsed their hands for 15 seconds under a stream of tap water. The test results are reported below in the following Table 14 which reports the relative performance of the E11 composition applied as indicated above, against the reference soft soap which was however applied in an amount of 5 ml, onto wet hands and handwashing continued for 60 seconds prior to rinsing in a stream of tap water for 15 seconds, thus having double the contact time with the hands than the E1 composition.

TABLE-US-00018 (comparative) (example composition) soft soap E11 person Log x Log y Log z Log x Log y Log z 1 7.02 3.97 3.05 6.66 3.51 3.15 2 6.77 3.65 3.12 6.98 3.30 3.68 3 6.51 3.73 2.78 6.30 3.36 2.94 4 6.96 3.60 3.36 6.80 3.13 3.67 5 6.84 3.49 3.35 6.87 3.08 3.79 6 6.37 3.78 2.59 6.70 3.76 2.94 7 6.22 3.07 3.15 6.22 2.99 3.23 8 6.99 3.72 3.27 6.83 3.06 3.77 9 6.01 3.40 2.61 6.36 3.00 3.36 10 6.87 3.74 3.13 7.04 3.64 3.40 11 6.91 3.62 3.29 6.78 3.09 3.69 12 7.04 3.94 3.10 6.80 3.09 3.71 13 6.33 3.46 2.87 6.29 2.94 3.35 14 6.64 3.66 2.98 6.96 2.93 4.03 15 6.73 3.54 3.19 6.74 2.91 3.83 X(m) 6.68 3.62 3.06 6.69 3.19 3.50 S 0.32 0.22 0.25 0.27 0.27 0.33 S 0.09 0.06 0.07 0.07 0.07 0.99 N 15 15 15 15 15 15 Log x = log of pre-count value/1 ml TSB Log y = log of post-count value/1 ml TSB Log z = log of reduction factor X(m) = mean S = standard deviation S = standard deviation of the mean N = number of subjects in the test

[0197] As is evident from the reported results of Table 14, the mean Log.sub.10 reduction of the challenge microorganism (reported as Log z on Table 14) was superior to that of the mean Log.sub.10 reduction of the soft soap.

[0198] While the invention is susceptible of various modifications and alternative forms, it is to be understood that specific embodiments thereof have been shown by way of example which are not intended to limit the invention to the particular forms disclosed; on the contrary the intention is to cover all modifications, equivalents and alternatives falling within the scope and spirit of the invention as expressed in the appended claims.