ANTIMICROBIAL MIXTURE CONTAINING 4-(3-ETHOXY-4-HYDROXYPHENYL)BUTAN-2-ONE AND AN ALCOHOL COMPOUND, AND COSMETIC COMPOSITION CONTAINING SAME

Abstract

The invention relates to an antimicrobial mixture containing 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one and an alcohol compound chosen from hydroxyacetophenone, 5-chloro-2-(2,4-dichlorophenoxy)phenol, sorbitan caprylate, the combination of 80% to 90% by weight, relative to the total weight of said combination, of xylityl sesquicaprylate and from 10% to 20% by weight of anhydroxylitol, 4-chloro-3,5-dimethylphenol, and glyceryl caprylate, and also to a cosmetic, pharmaceutical or nutritional composition containing such a mixture.

Application to caring for, making up and cleansing keratin materials; to preserving foods and to water treatment.

Claims

1. An antimicrobial mixture comprising: one or more 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one, salt thereof with an organic or inorganic acid or base, solvate thereof and mixture thereof; and one or more alcohol compounds chosen from: hydroxyacetophenone and salts thereof with an organic or inorganic base, and solvates thereof; 5-chloro-2-(2,4-dichlorophenoxy)phenol and salts thereof with an organic or inorganic base, and solvates thereof; sorbitan caprylate, and its optical isomers, and solvates thereof; the combination of 80% to 90% by weight, relative to the total weight of said combination, of xylityl sesquicaprylate and its optical isomers, and solvates thereof and from 10% to 20% by weight of anhydroxylitol and its optical isomers, and solvates thereof; 4-chloro-3,5-dimethylphenol and salts thereof with an organic or inorganic base, and solvates thereof; and glyceryl caprylate and its optical isomers, and solvates thereof.

2. The mixture as claimed in claim 1, wherein the alcohol compound is hydroxyacetophenone, salt thereof with an organic or inorganic base, solvate thereof and mixture thereof.

3. The mixture as claimed in claim 1, which has a 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one/hydroxyacetophenone weight ratio ranging from 0.08 to 0.5.

4. The mixture as claimed in claim 1, wherein the alcohol compound is 5-chloro-2-(2,4-dichlorophenoxy)phenol, salt thereof with an organic or inorganic base, salt thereof and mixture thereof.

5. The mixture as claimed in claim 4, which has a 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one/5-chloro-2-(2,4-dichlorophenoxy)phenol weight ratio ranging from 0.3 to 6.

6. The mixture as claimed in claim 1, wherein the alcohol compound is sorbitan caprylate, an optical isomer thereof, solvate thereof and mixture thereof.

7. The mixture as claimed in claim 6, which has a 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one/sorbitan caprylate weight ratio ranging from 0.1 to 3.2.

8. The mixture as claimed in claim 6, which has a 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one/sorbitan caprylate weight ratio ranging from 0.4 to 1.6.

9. The mixture as claimed in claim 6, which has a 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one/sorbitan caprylate weight ratio ranging from 0.1 to 3.2.

10. The mixture as claimed in claim 1, wherein the alcohol compound is a combination of from 80% to 90% by weight, relative to the total weight of said combination, of xylityl sesquicaprylate, an optical isomer thereof, solvate thereof and mixture thereof and from 10% to 20% by weight of anhydroxylitol, an optical isomer thereof, solvate thereof and mixture thereof.

11. The mixture as claimed in claim 10, which has a 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one/combination of xylityl sesquicaprylate/anhydroxylitol weight ratio ranging from 0.2 to 13.

12. The mixture as claimed in claim 10, which has a 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one/combination of xylityl sesquicaprylate/anhydroxylitol weight ratio ranging from 0.4 to 3.2.

13. The mixture as claimed in claim 1, wherein the alcohol compound is 4-chloro-3,5-dimethylphenol, salt thereof with an organic or inorganic base, solvate thereof and mixture thereof.

14. The mixture as claimed in claim 13, which has a 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one/4-chloro-3,5-dimethylphenol weight ratio ranging from 0.1 to 1.4.

15. The mixture as claimed in claim 13, which has a 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one/4-chloro-3,5-dimethylphenol weight ratio ranging from 0.15 to 1.4.

16. The mixture as claimed in claim 13, which has a 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one/4-chloro-3,5-dimethylphenol weight ratio ranging from 0.1 to 0.7.

17. The mixture as claimed in claim 1, wherein the alcohol compound is glyceryl caprylate, an optical isomer thereof, solvate thereof and mixture thereof.

18. The mixture as claimed in claim 17, which has a 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one/glyceryl caprylate weight ratio ranging from 0.1 to 10.

19. The mixture as claimed in claim 17, which has a 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one/glyceryl caprylate weight ratio ranging from 0.5 to 9.

20. The mixture as claimed in claim 17, which has a 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one/glyceryl caprylate weight ratio ranging from 0.2 to 5.

21. A composition in a physiologically acceptable medium, and comprising the mixture as claimed in claim 1.

22. A composition comprising, in a physiologically acceptable medium, an antimicrobial mixture as claimed in claim 1.

23. The composition as claimed in claim 21, which comprises at least one additional ingredient chosen from water, oils, polyols containing from 2 to 10 carbon atoms, gelling agents, surfactants, film-forming polymers, dyestuffs, fragrances, fillers, UV-screening agents, plant extracts, cosmetic and dermatological active agents, and salts.

24. The composition as claimed in claim 21, wherein the 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one, salt thereof with an organic or mineral base, solvate thereof and mixture thereof is present in a content ranging from 0.01% to 5% by weight relative to the total weight of the composition.

25. A nontherapeutic cosmetic treatment process for caring for and/or making up and/or cleansing keratin materials, comprising the application to said keratin materials of a composition as claimed in claim 21.

26. A process for conserving a composition, comprising a physiologically acceptable medium which comprises incorporating into said composition an antimicrobial mixture as defined in claim 1.

27. A process for preserving a nutritional composition, which comprises incorporating into said composition an antimicrobial mixture as defined in claim 1.

28. A process for treating water which comprises incorporating an antimicrobial mixture as defined in claim 1, into the water, wherein the water is chosen from domestic or industrial waters, waters from aquatic media, swimming pool/spa waters, and waters from air-conditioning systems.

29. A continuous or discontinuous water treatment process comprising at least one step of placing a water sample to be treated or a water stream to be treated, said water being chosen from domestic or industrial waters, waters from aquatic media, swimming pool/spa waters, and waters from air-conditioning systems, in contact with the antimicrobial mixture as defined in claim 1.

Description

EXAMPLES

Example 1: Determination of the Synergistic Antimicrobial Activity as MIC

[0139] The demonstration of a synergistic antimicrobial activity effect with a mixture of 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one (referred to as substance A) and of an alcohol compound (referred to as substance B) was performed by calculating the synergy index (or FIC index) according to the following formula:

FIC Index=(MIC of A with B/MIC of A)+(MIC of B with A/MIC of B  [Math. 1]

[0140] with: [0141] MIC of A with B: minimum concentration of product A in the combination A+B which makes it possible to obtain an inhibitory effect; [0142] MIC of B with A: minimum concentration of product B in the combination A+B which makes it possible to obtain an inhibitory effect; [0143] MIC of A: minimum inhibitory concentration of product A alone; [0144] MIC of B: minimum inhibitory concentration of product B alone.
This formula was described for the first time in the article by F. C. Kull, P. C. Eisman, H. D. Sylwestrowka, and R. L. Mayer, Applied Microbiology 9:538-541, 1961.

[0145] For each compound tested alone, the MIC is considered as the first concentration which makes it possible to obtain a microbial growth percentage of less than or equal to 25%.

[0146] As regards the combinations tested, MIC of A with B and MIC of B with A are the respective concentrations of A and of B in the combinations which make it possible to obtain a microbial growth percentage of less than or equal to 25%.

[0147] Interpretation of the FIC Index:

[0148] When the FIC index value is less than or equal to 1, it is considered that the combination of test compounds has a synergistic effect.

[0149] The summary of the results obtained is presented in the following tables.

[0150] The combination of compounds A and B was tested on the following strains or a part thereof: Aspergillus niger, Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecalis, Candida albicans.

[0151] The microbial strain Aspergillus niger ATCC 6275, and a double-concentration Sabouraud broth liquid culture medium supplemented with polyoxyethylenated (20 OE) sorbitan monopalmitate (Tween 40 from Croda) and Phytagel© BioReagent were used (i.e. a mixture of 5 g of Phytagel+0.6 g of Tween 40+60 g of Sabouraud broth).

[0152] The microbial strain Staphylococcus aureus ATCC 6538 and a double-concentration nutrient broth liquid culture medium were used.

[0153] The microbial strain Pseudomonas aeruginosa ATCC 9027 and a double-concentration nutrient broth liquid culture medium were used.

[0154] The microbial strain Enterococcus faecalis ATCC 33186 and a double-concentration BHI (Brain Heart Infusion) broth liquid culture medium were used.

[0155] The microbial strain Candida albicans ATCC 10231, and a double-concentration Sabouraud broth liquid culture medium were used (i.e. a mixture of 5 g of Phytagel+0.6 g Tween 40+60 g of Sabouraud broth).

[0156] A 96-well microplate at an incubation temperature of 32.5° C. is used.

[0157] The incubation time of the microplate is: [0158] from 24 to 30 h under aerobic conditions for microbial Aspergillus niger ATCC 6275; [0159] from 18 to 24 h under aerobic conditions for Candida albicans ATCC 10231, Pseudomonas aeruginosa ATCC 9027 and Staphylococcus aureus ATCC 6538; [0160] from 24 to 48 h under aerobic conditions for Enterococcus faecalis ATCC 33186.

[0161] Tests

For each compound:
A=4-(3-ethoxy-4-hydroxyphenyl) butan-2-one compound
B=alcohol compound
A 10% (weight/volume) stock solution was prepared by mixing 1 g of compound in 9 ml of aqueous 1% agar solution. Successive dilutions were made with the 1% agar solution.

[0162] Tests of Compounds A and B Alone

[0163] 50 μl of each of the daughter solutions obtained containing compound A or B are added to the microplate wells. 100 μl of Sabouraud liquid nutrient broth inoculated at double concentration with the Aspergillus niger strain and 50 μl of aqueous 1% agar solution are also added thereto.

[0164] Tests of Compounds A and B as a Mixture

[0165] 50 μl of each of the daughter solutions obtained containing compound A and 50 μl of each of the daughter solutions obtained containing compound B are added to the microplate wells. 100 μl of Sabouraud liquid nutrient broth inoculated at double concentration with the strain Aspergillus niger are also added thereto.

[0166] Microbial Growth Control

[0167] A positive microbial growth control was also prepared. The positive microbial growth control corresponds to a mixture of 100 μl of aqueous 1% agar solution with 100 μl of Sabouraud liquid nutrient broth inoculated at double concentration with the strain Aspergillus niger in the absence of compounds A and B.

[0168] Absorbance Control for Compounds A and B Alone

[0169] An absorbance control was performed in parallel on compounds A and B alone. This control corresponds to 100 μl of double concentration sterile Sabouraud liquid nutrient broth+100 μl of double concentration compound A or B.

[0170] In the three cases (absorbance control, growth control and test), the final volume present in each of the microplate wells is 200 μl.

[0171] In the two cases (test and control), the inoculum represents the concentration of the Aspergillus niger strain present in the final volume of the wells (200 μl) and is between 2 and 6×10.sup.5 cfu/ml of Aspergillus niger.

[0172] The minimum inhibitory concentration (MIC) of each compound A and B alone and in combination was determined in a known manner by means of optical density measurements at a wavelength of 620 nm.

[0173] The test as described above (tests, absorbance control and growth control) was performed again to test the combination A+B on the following strains Enterococcus faecalis, Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, where appropriate.

[0174] The following results were obtained with compound B1=4-hydroxyacetophenone:

[0175] Aspergillus niger

TABLE-US-00001 TABLES 1 Concentrations tested (in weight %) 0 A 0.025 A 0.05 A 0.1 A 0 B1 82 43 5 0.0625 B1 86 71 46 3 0.125 B1 72 73 20 2 (FIC 0.75) 0.25 B1 79 22 12 5 (FIC 0.75) (FIC 1) 0.5 B1 6 2 −19 6 (FIC 0.75) (FIC 1)

TABLE-US-00002 TABLES 2 MIC of each compound as a % MIC of % MIC of mixture FIC Ratio A alone B1 alone A% B1 % Index A/B1 0.1 0.5 0.05 0.25 1 0.2

[0176] The results obtained show synergistic inhibitory activity for the mixtures:

0.05% of A and 0.25% of B1, i.e. A/B1 ratio=0.2
0.05% of A and 0.125% of B1, i.e. A/B1 ratio=0.4
0.025% of A and 0.25% of B1, i.e. A/B1 ratio=0.1

Example 2: Determination of the Synergistic Antimicrobial Activity as MIC on Staphylococcus aureus

[0177] The demonstration of a synergistic antimicrobial activity effect with a mixture of 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one (referred to as substance A) and of 5-chloro-2-(2,4-dichlorophenoxy)phenol (referred to as substance B2) was performed according to the conditions described in Example 1.

[0178] The following results were obtained:

[0179] Staphylococcus aureus

TABLE-US-00003 TABLES 3 Concentrations tested (in weight %) 0 A 0.0625 A 0.125 A 0.25 A 0.5 A 1 A 0 B2 52 49 52  75  2 0.125 B2 46 −8 −4 −9 −12 −1 (FIC (FIC (FIC (FIC 0.56) 0.63) 0.75) 1) 0.25 B2 14 −7  6  5 −10 −1

TABLE-US-00004 TABLES 4 MIC of each compound as % MIC of A % MIC of B2 a mixture FIC Ratio alone alone A% B2% Index A/B2 1 0.25 0.5 0.125 1 4

[0180] The results obtained show synergistic inhibitory activity for the mixtures:

i) 0.5% of A and 0.125% of B2, i.e. A/B2 ratio=4
ii) 0.25% of A and 0.125% of B2, i.e. A/B2 ratio=2
iii) 0.125% of A and 0.125% of B2, i.e. A/B2 ratio=1
iv) 0.0635% of A and 0.125% of B2, i.e. A/B2 ratio=0.5

Example 3

[0181] The demonstration of a synergistic antimicrobial activity effect with a mixture of 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one (referred to as substance A) and of sorbitan caprylate (referred to as substance B3) was performed according to the conditions described in Example 1.

[0182] Aspergillus niger

TABLE-US-00005 TABLES 5 Concentrations tested 0 A 0.025 A 0.05 A 0.1 A (in weight %) 0 B3 45 35 14 0.025 B3 50 42 26 12 0.05 B3 30 19 15 5 (FIC 0.75) (FIC 1) 0.1 B3 −2 3 8 6

TABLE-US-00006 TABLES 6 % MIC % MIC MIC of each compound of A of B3 as a mixture FIC Ratio alone alone A % B3 % Index A/B3 0.1 0.1 0.05 0.05 1 1

[0183] The results obtained show synergistic inhibitory activity for the mixtures:

i) 0.05% of A and 0.05% of B3, i.e. A/B3 ratio=1
ii) 0.025% of A and 0.05% of B3, i.e. A/B3 ratio=0.5

[0184] Enterococcus faecalis

TABLE-US-00007 TABLES 7 Concentrations tested 0 A 0.0625 A 0.125 A 0.25 A 0.5 A (in weight %) 0 B3 70 31 27 14 0.125 B3 134 117 62 −16 −1 0.25 B3 61 −85 4 2 4 0.5 B3 62 −8 0 −9 −12 1 B3 2 −8 −5 −1 3

TABLE-US-00008 TABLES 8 % MIC % MIC MIC of each compound of A of B3 as a mixture FIC Ratio alone alone A % B3 % Index A/B3 0.5 1 0.25 0.5 1 0.5

[0185] The results obtained show synergistic inhibitory activity for the mixtures:

[0186] i) 0.25% of A and 0.5% of B3, i.e. A/B3 ratio=0.5

[0187] ii) 0.125% of A and 0.5% of B3, i.e. A/B3 ratio=0.25

[0188] iii) 0.0625% of A and 0.5% of B3, i.e. A/B3 ratio=0.125

[0189] iv) 0.25% of A and 0.25% of B3, i.e. ratio A/B3=1

[0190] v) 0.125% of A and 0.25% of B3, i.e. ratio A/B3=0.5

[0191] vi) 0.0625% of A and 0.25% of B3, i.e. ratio A/B3=0.25

[0192] vii) 0.25% of A and 0.125% of B3, i.e. ratio A/B3=2

Example 4

[0193] The demonstration of a synergistic antimicrobial activity effect with a mixture of 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one (referred to as substance A) and of the combination of xylityl sesquicaprylate and anhydroxylitol (85/15 weight/weight) sold under the name Hebeatol Plus from Chemyunion (referred to as substance B4) was performed according to the conditions described in Example 1.

[0194] Pseudomonas aeruginosa

TABLE-US-00009 TABLES 9 Concentrations tested (in weight %) 0 A 0.0625 A 0.125 A 0.25 A 0.5 A 1 A 0 B4 63 60 64  62  7 0.0625 B4 100 60 27 12 −10 −8 (FIC (FIC 0.38) 0.5) 0.125 B4  57 15 −4 −9  3  0 (FIC (FIC (FIC (FIC 0.31) 0.38) 0.5) 0.75) 0.25 B4  52 −5 −9 −23  −22 −5 (FIC (FIC (FIC (FIC 1) 0.56) 0.63) 0.75) 0.5 B4  20 −5 −13  −11   7 −3

TABLE-US-00010 TABLES 10 % MIC % MIC MIC of each compound of A of B4 as a mixture FIC Ratio alone alone A % B4 % Index A/B4 1 0.5 0.5 0.25 1 2

[0195] The results obtained show synergistic inhibitory activity for the mixtures:

i) 0.5% of A and 0.25% of B4, i.e. A/B4 ratio=2
ii) 0.25% of A and 0.25% of B4, i.e. A/B4 ratio=1
iii) 0.125% of A and 0.25% of B4, i.e. A/B4 ratio=0.5
iv) 0.0625% of A and 0.25% of B4, i.e. A/B4 ratio=0.25
v) 0.5% of A and 0.125% of B4, i.e. ratio A/B4=4
vi) 0.25% of A and 0.125% of B4, i.e. ratio A/B4=2
vii) 0.125% of A and 0.125% of B4, i.e. ratio A/B4=1
viii) 0.0625% of A and 0.125% of B4, i.e. ratio A/B4=0.5
ix) 0.5% of A and 0.0625% of B4, i.e. ratio A/B4=8
x) 0.25% of A and 0.0625% of B4, i.e. ratio A/B4=4

[0196] Enterococcus faecalis

TABLE-US-00011 TABLES 11 Concentrations tested 0 A 0.0625 A 0.125 A 0.25 A 0.5 A (in weight %) 0 B4 44 39 38 12 0.0625 B4 45 53 41 30 5 0.125 B4 42 6 7 1 11 (FIC 0.63) (FIC 0.75) (FIC 1) 0.25 B4 −6 −10 1 −4 −4

TABLE-US-00012 TABLES 12 % MIC % MIC MIC of each compound of A of B4 as a mixture FIC Ratio alone alone A % B4 % Index A/B4 0.5 0.25 0.25 0.125 1 0.5

[0197] The results obtained show synergistic inhibitory activity for the mixtures:

i) 0.25% of A and 0.125% of B3, i.e. A/B4 ratio=2
ii) 0.125% of A and 0.125% of B3, i.e. A/B4 ratio=1
iii) 0.0625% of A and 0.125% of B3, i.e. A/B4 ratio=0.5

Example 5

[0198] The demonstration of a synergistic antimicrobial activity effect with a mixture of 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one (referred to as substance A) and of 4-chloro-3,5-dimethylphenol (referred to as substance B5) was performed according to the conditions described in Example 1.

[0199] Candida Albicans

TABLE-US-00013 TABLES 13 Concentrations tested 0 A 0.025 A 0.05 A 0.1 A 0.2 A (in weight %) 0 B5 69 54 27 4 0.0625 B5 73 72 48 22 3 0.125 B5 28 15 11 6 1 (FIC 0.63) (FIC 0.75) (FIC 1) 0.25 B5 1 0 0 1 0

TABLE-US-00014 TABLES 14 % MIC % MIC MIC of each compound of A of B5 as a mixture FIC Ratio alone alone A % B5 % Index A/B5 0.2 0.25 0.1 0.125 1 0.8

[0200] The results obtained show synergistic inhibitory activity for the mixtures:

0.1% of A and 0.125% of B5, i.e. A/B5 ratio=0.8
0.05% of A and 0.125% of B, i.e. A/B5 ratio=0.4
0.025% of A and 0.125% of B, i.e. A/B5 ratio=0.2

[0201] Aspergillus niger

TABLE-US-00015 TABLES 15 Concentrations tested 0 A 0.025 A 0.05 A 0.1 A (in weight %) 0 B5 55 32 16 0.0625 B5 99 31 21 13 0.125 B5 103 40 7 7 (FIC 0.75) 0.25 B5 31 48 2 7 (FIC 1) 0.5 B5 −6 −27 −8 −6

TABLE-US-00016 TABLES 16 % MIC % MIC MIC of each compound of A of B5 as a mixture FIC Ratio alone alone A % B5 % Index A/B5 0.1 0.5 0.05 0.25 1 0.2
The results obtained show synergistic inhibitory activity for the mixtures:
i) 0.05% of A and 0.25% of B5, i.e. A/B5 ratio=0.2
ii) 0.05% of A and 0.125% of B5, i.e. A/B5 ratio=0.4

Example 6

[0202] The demonstration of a synergistic antimicrobial activity effect with a mixture of 4-(3-ethoxy-4-hydroxyphenyl)butan-2-one (referred to as substance A) and of glyceryl caprylate (referred to as substance B6) was performed according to the conditions described in Example 1.

[0203] Pseudomonas aeruginosa

TABLE-US-00017 TABLES 17 Concentrations tested (in weight %) 0 A 0.0625 A 0.125 A 0.25 A 0.5 A 1 A 0 B6 80 63 56 61 −17 0.1 B6 123 84 58 18 11  −7 (FIC (FIC 0.38) 0.75) 0.2 B6 109 44 10 −5 −1  −5 (FIC (FIC (FIC 1) 0.63) 0.75) 0.4 B6 −40 12 −5  2  1  −2

TABLE-US-00018 TABLES 18 % MIC % MIC MIC of each compound of A of B6 as a mixture FIC Ratio alone alone A % B6 % Index A/B6 1 0.4 0.5 0.2 1 2.5

[0204] The results obtained show synergistic inhibitory activity for the mixtures:

i) 0.5% of A and 0.2% of B6, i.e. A/B6 ratio=2.5
ii) 0.25% of A and 0.2% of B6, i.e. A/B6 ratio=1.25
iii) 0.125% of A and 0.2% of B6, i.e. A/B6 ratio=0.625
iv) 0.5% of A and 0.1% of B6, i.e. A/B6 ratio=5
v) 0.25% of A and 0.1% of B6, i.e. ratio A/B6=2.5

[0205] Staphylococcus aureus

TABLE-US-00019 TABLES 19 Concentrations tested (in weight %) 0 A 0.0625 A 0.125 A 0.25 A 0.5 A 1 A 0 B6 65 75 79 24  5 0.0625 B6 83 80 27 −1  1 −1 (FIC (FIC 0.31) 0.56) 0.125 B6 87 58  0  1 −9  0 (FIC (FIC (FIC 0.25) 0.38) 0.63) 0.25 B6 53 −4  1  7  1  0 (FIC (FIC (FIC (FIC 0.31) 0.38) 0.5) 0.75) 0.5 B6 48  0 −6  4  0 −1 (FIC (FIC (FIC (FIC 1) 0.56) 0.63) 0.75) 1 B6  9 −1  0  6 −3 −6

TABLE-US-00020 TABLES 20 % MIC % MIC MIC of each compound as a of A of B6 mixture FIC Ratio alone alone A % B6 % Index A/B6 1 1 0.5 0.5 1 1

[0206] The results obtained show synergistic inhibitory activity for the mixtures:

i) 0.5% of A and 0.5% of B6, i.e. A/B6 ratio=1
ii) 0.25% of A and 0.5% of B6, i.e. A/B6 ratio=0.5
iii) 0.125% of A and 0.5% of B6, i.e. A/B6 ratio=0.25
iv) 0.0625% of A and 0.5% of B6, i.e. ratio A/B6=0.125
iv) 0.5% of A and 0.25% of B6, i.e. A/B6 ratio=2
v) 0.25% of A and 0.25% of B6, i.e. ratio A/B6=1
vi) 0.125% of A and 0.25% of B6, i.e. ratio A/B6=0.5
vii) 0.0625% of A and 0.25% of B6, i.e. ratio A/B6=0.25
viii) 0.5% of A and 0.125% of B6, i.e. ratio A/B6=4
ix) 0.25% of A and 0.125% of B6, i.e. ratio A/B6=2
x) 0.125% of A and 0.125% of B6, i.e. ratio A/B6=1
xi) 0.5% of A and 0.0625% of B6, i.e. ratio A/B6=8
xii) 0.25% of A and 0.0625% of B6, i.e. ratio A/B6=4

[0207] E. Faecalis

TABLE-US-00021 TABLES 21 Concentrations tested 0 A 0.0625 A 0.125 A 0.25 A 0.5 A (in weight %) 0 B6 35 43 34 0 0.625 B6 38 16 12 4 −4 (FIC (FIC (FIC 0.25) 0.38) 0.63) 0.125 B6 57 5 −1 −1 −7 (FIC (FIC 0.5) (FIC 0.38) 0.75) 0.25 B6 22 −19 −12 −11 3 (FIC (FIC (FIC 1) 0.63) 0.75) 0.5 B6 8 −3 1 −24 −7

TABLE-US-00022 TABLES 22 % MIC % MIC MIC of each compound of A of B6 as a mixture FIC Ratio alone alone A % B6 % Index A/B6 0.5 0.5 0.25 0.25 1 1

[0208] The results obtained show synergistic inhibitory activity for the mixtures:

i) 0.25% of A and 0.25% of B6, i.e. A/B6 ratio=1
ii) 0.125% of A and 0.25% of B6, i.e. A/B6 ratio=0.5
iii) 0.0625% of A and 0.25% of B6, i.e. A/B6 ratio=0.25
iv) 0.25% of A and 0.125% of B6, i.e. ratio A/B6=2
v) 0.125% of A and 0.125% of B6, i.e. ratio A/B6=1
vi) 0.0625% of A and 0.125% of B6, i.e. ratio A/B6=0.5
vii) 0.25% of A and 0.0625% of B6, i.e. ratio A/B6=4
viii) 0.125% of A and 0.0625% of B6, i.e. ratio A/B6=2
ix) 0.0625% of A and 0.0625% of B6, i.e. ratio A/B6=1