PL181504B1

Water-soluble synergistic protective compositions of lethal compounds

Abstract

A water soluble preservative admixture of biocidal compounds for addition to commercial use compositions at predetermined use levels, and uniformly distributed therein, to provide long-time synergistic biocidal activity against a wide range of fungi and both gram-negative and gram-positive bacteria, which comprises powders of (a) one or more methylol compounds, or their equivalents, and (b) iodopropynyl alcohol, or its ester, carmabate or ether derivative thereof, in a weight ratio of (a):(b) of 100:1 to 2000:1.

Term

No projected expiry on record.

7 claims: 2 independent, 5 dependent

1
Patent claims Zastrzeżenia patentowe 1. A water-soluble, antimicrobial protective composition, being an addition to commercial preparations, showing synergism against a wide spectrum of fungi and bacteria, both Gram-negative and Gram-positive, containing a mixture (1):1. Rozpuszczalna w wodzie, przeciwbakteryjna kompozycja ochronna, stanowiąca dodatek do handlowych preparatów użytkowych, wykazująca synergizm przeciwko szerokiemu spektrum grzybów i bakterii, zarówno Gram-ujemnych jak i Gram-dodatnich, zawierająca mieszaninę (1): (a) N- [1,3-bis (hydroxymethyl) -2,5-dioxo-4-imidazoiidinyl] -N, N'-bis- (hydroxymethyl) urea or 1,3-dihydroxymethyl-5,5-dimethylhydantoin, or dihydroxymethylglycine anhydride, and (b) 3-iodo-2-propynyl butylcarbamate, characterized in that the weight ratio of components (a): (b) is from 100: 1 to 2000: 1, and that the composition optionally comprises the component (2) glycol or 1,3-butylene glycol, the amount of the mixture (1) in the composition being from 20% to 55% by weight, and the amount of component (2) is from 45% to 80% by weight. (a) N-[ 1,3-bis(hydroksymetylo)-2,5-diokso-4-imidazoiidynylo]-N,N'-bis-(hydroksymetylo)moczmka lub 1,3-dihydroksymetylo-5,5-dimetylohydantoiny, lub bezwodnika dihydroksymetyloglicyny, oraz (b) butylokarbaminianu 3-jodo-2-propynylu, znamienna tym, że stosunek wagowy składników (a): (b) wynosi od 100:1 do 2000:1, oraz że kompozycja ewentualnie zawiera składnik (2) glikol lub glikol 1,3-butylenowy, przy czym ilość mieszaniny (1) w kompozycji wynosi od 20% do 55% wagowych, zaś ilość składnika (2) wynosi od 45% do 80% wagowych.
4
A commercial commercial preparation for cosmetic and economic purposes, with a prolonged period of resistance to fungi and bacteria, characterized in that it contains from 0.01% to 5% by weight of a water-soluble protective composition containing (1) a mixture of:4. Użytkowy preparat handlowy do celów kosmetycznych i gospodarczych, o przedłużonym okresie odporności na grzyby i bakterie, znamienny tym, że zawiera od 0,01% do 5% wagowych rozpuszczalnej w wodzie kompozycji ochronnej zawierającej (1) mieszaninę: (a) N- [1,3-bis (hydroxymethyl) -2,5-dioxo-4 ^ -imK ^ £ ^ o ^ ^ ^ ^ ^ ^ ^ k3] N, N '' - hii - (t ^ i ^ t ^ i ^ c ^ k ^ sym ^ ethyl) urea or 1 J-dihydroxymethyl GP-dimethylhydantomy, or dihydroxymethylglycine anhydride, and (b) 3-cyclo-2-prcpyryl butylcarbamate, by weight of the components (a ): (b) from 100: 1 to 2000: 1, and optionally containing (2) propylene glycol or 1,3-butylene glycol, the amount of the mixture (1) in the composition being 20% to 55% by weight, and the amount component (2) is from 45% to 80% by weight. (a) N-[ 1,3-bis(hydroksymetylo)-2,5-diokso-4^-imK^£^z^oik^}^r^y^'k3]N,N''-hii:'-(ł^>^t^i^c^k^sym^etylo)mocznika lub 1 J-dihydroksymetyloGp-dimetylohydantomy, lub bezwodnika dihydroksymetyloglicyny, oraz (b) butylckarbamimanu 3-jcdo-2-prcpyrylu, w stosunku wagowym składników (a) : (b) wynoszącym od 100:1 do 2000:1, oraz ewentualnie zawierającej składnik (2) glikol propylenowy lub glikol 1,3-butylenowy, przy czym ilość mieszaniny (1) w kompozycji wynosi od 20% do 55% wagowych, zaś ilość składnika (2) wynosi od 45% do 80% wagowych.

Independent claims2

1,434 paragraphs, as filed

The subject of the invention is a water-soluble protective composition admixing commercial commercial preparations in order to obtain a long period of their synergistic life-killing activity, and a commercial commercial preparation containing this composition. More specifically, the invention relates to a mixture of certain compounds containing a hydroxymethyl group and a compound containing an i-propopyryl group.

In cict3 ^ (^^ h ^; zαsccwycb studies, combinations of antibacterial agents were developed to:

1) obtaining biochemical synergism,

2) extending the range of antibacterial activity of each agent,

3) increasing the solubility of the mixture in water,

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4) minimizing the toxicity or irritation of the host to the host, and

5) minimizing physical and chemical incompatibility.

Real biological synergism exists when two agents, when combined, require the use of smaller amounts to produce approximately the same inhibitory or lethal effect than each of these individual agents alone. Although the synergistic interaction of two or more antibacterial agents produces a greater effect than the cumulative effect of biological activity, in most cases the mechanism of this synergism is unknown.

For example, M. Rosen et al. Disclose in US Patent No. 4,844891 a protective composition comprising (a) a formaldehyde donor, containing a hydroxymethyl group, and (b) a halopropynyl compound, in a weight ratio (a): (b) of from 50: 1 to 1: 1, preferably from 30: 1 to 2: 1 and most preferably from 20: 1 to 10: 1, which composition allows the fungicidal activity of commercial preparations to be maintained for 1-3 days. However, Rosen observed that when the ratio (a): (b) in the concentrate is greater than 50 1 (System No. 16 in Table 1, ratio 73.33), the protective composition was ineffective from the point of view of preserving the life-killing properties of the preparations used . Thus, Rosen needed a relatively large amount of the halopyropinyl compound to achieve significant lethal activity in the compositions used. In such compositions, although the formaldehyde donor is water soluble, the halopyropinyl compound is substantially water insoluble. Therefore, Rosen had difficulty in uniformly dispersing his composition in the formulation.

For these and other reasons, it is desirable to provide a new and improved, water-soluble protective composition of such life-killing compounds, which requires the use of a relatively small amount of water-insoluble and expensive halogen-propopynyl compound, and which also allows effective synergistic protection of formulations. spectrum of fungi and bacteria at various levels of use and for a long period of time.

It is an object of the present invention to provide a concentrate composition that is water-soluble and which can thus be uniformly dispersed in formulations at a predetermined level of use.

Another object of the present invention is to provide a protective composition that provides a long period of synergistic life-killing activity in formulations, against a broad spectrum of fungi and bacteria, both gram-positive and gram-negative.

Still another object of the present invention is to provide a water-soluble protective composition for personal care formulations in the form of a solution, dishwashing liquid, gel, emulsion, emulsifiable concentrate, suspension, slurry or cream.

These and other features and features of the invention will be apparent from the more detailed description below.

The subject of the invention is a water-soluble protective composition, which is an addition to commercial utility preparations, with predetermined levels of use and uniformly dispersed in these preparations, which composition provides a long period of synergistic activity against a wide spectrum of fungi and bacteria, both Gram-negative and Gram-positive bacteria.

The composition according to the invention consists of (1) a mixture of powders:

(a) N- [1,3-bis (hydroxymethyl) -2,5-dioxo-4-imidazolidinyl] -N, N'-bis- (hydroxymethyl) urea or 1,3-dihydroxymethyl-5,5-dimethylhydantoin, or dihydroxymethylglycine anhydride, and (b) 3-iodo-2-propynyl butylcarbamate, and is characterized in that the weight ratio of components (a): (b) is from 100: 1 to 2000: 1.

Preferably the composition comprises as a second component (2) propylene glycol or glycol

1,3-butylene, the amount of the mixture (1) in the composition being from 20% to 55% by weight, and the amount of component (2) being from 45% to 80% by weight.

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Preferably, in the protective composition of the invention, the weight ratio of components (a): (b) is from 200: 1 to 500: 1.

Preferably, the protective composition contains 40% by weight of component (1) and 60% by weight of component (2).

The subject of the invention is also a commercial commercial preparation with a prolonged period of resistance to contamination by a wide spectrum of Gram-negative and Gram-positive bacteria and characterized by containing from 0.01% to 5% by weight of a water-soluble protective composition containing (1) mixture:

(a) N- [1,3-bis (hydroxymethyl) -2,5-dioxo-4-imidazolidinyl] -N, N'-biS ((hydrc) xymethyl) urea, or 1,3-dihydroxymethyl-5.5 -dimethylhydantoin or dihydroxymethylglycine anhydride, and (b) 3-iodo-2-propynyl butylcarbamate, in a weight ratio of components (a) :( b) of 100.1 to 2000: 1, and optionally containing component (2) propylene glycol or glycol 1,3-butylene, the amount of mixture (1) in the composition being from 20% to 55% by weight, and the amount of component (2) is from 45% to 80% by weight.

Preferably, the commercial use formulation according to the invention contains 0.1% by weight of a water-soluble protective composition.

Preferably, the commercial formulation contains the composition dissolved in water and uniformly dispersed in the formulation.

Preferably, the content of component (b) in the formulation ranges from 0.5 ppm to 10 ppm, and the content of component (a) in the formulation is at least 250 ppm.

Preferably, the formulation according to the invention contains from 0.5% to 1% by weight of the composition.

The commercial commercial preparation according to the invention may be a personal care preparation for household or industrial purposes.

The commercial use formulations of the present invention contain from 0.01% to 2% by weight of the composition of the invention. Such formulations contain a compound with an iodopropynyl group in an amount of 0.5 ppm to 10 ppm to provide the desired antifungal activity, and a compound containing a hydrate group, or an equivalent thereof. at least 250 ppm to provide the desired antibacterial activity.

The invention is based on discoveries that, in a predetermined mixture consisting of (a) a hydroxymethyl containing compound or equivalent and (b) an iodopropynyl containing compound:

1) Compounds containing an iodopropynyl group, with a weight ratio of components (a) :( b) less than 100, are largely insoluble in water and, respectively, with a weight ratio below 100: 1, it is difficult to uniformly distribute the compound containing the iodopropynyl group in preparations in which are used water, especially in creams, gels and the like. According to the invention, the compositions are used at weight ratios of the components (a) :( b) from 100: 1 to 2000: 1, which are water-soluble in all formulations at normal levels of use.

2) Effective, synergistic life-killing activity is obtained with compositions in which the weight ratio of components (a) :( b) is from 100: 1 to 2000: 1. The value of the synergistic index (SI) of such mixtures approaches zero (maximum synergism) in relation to a wide spectrum of microorganisms. In contrast, mixtures in which the ratios (a) :( b) are below 100: 1, for example from 10: 1 to 50: 1, show much less synergistic properties and are only active on a narrower spectrum of microorganisms

3) The most effective protective activity in relation to the use preparations is obtained with compositions in which the weight ratio is from 100: 1 to 2000: 1, with levels of use from 0.05% to 2% by weight of the final product. At these values, the amount of iodopropynyl group-containing compound is only from 0.5 ppm to 10 ppm, which significantly reduces the cost as well as the toxicity of the use composition. The hydroxymethyl group containing compound is also present in an amount of at least 250 ppm.

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The experimental results on which these discoveries are based are presented below. In these examples, the hydrocymethyl group-containing compound (a) can be selected from (GERMALL® II) N- [1,3-bis (hycyroxynethyl) -2,5-dioxo-4-imidazolidinyl] -N, N'-bis (hydroxymethyl) urea, (DMDMH) 1,3-dihydroxymethyl-5,5-dimethylhydantoin, and (GADM) dihydroxymethylglycine anhydride, while the compound (b) containing the iodopropynyl group is iodopropynyl alcohol (IPGA) or 3-iodo-2-propynyl butylcarbamate (IPBC)

1. Water solubility

The water solubility or insolubility of compositions constituting a mixture of a number of compounds containing a hydroxymethyl group from IPBC in the form of 1% aqueous solutions is presented in Tables A to C below.

Table A.

<td>GERMALL® II.IPBC weight ratio</td><td>Quantity in ppm</td><td>Solubility</td>

<td> 2000 1</td><td> 5</td><td>soluble</td>

<td> 1000 1</td><td> 10</td><td>soluble</td>

<td> 500 1</td><td> 20</td><td>soluble</td>

<td> 200 1</td><td> 50</td><td>soluble</td>

<td> 100 1</td><td> 100</td><td>soluble</td>

<td> 50 1</td><td> 200</td><td>insoluble</td>

<td> 20-1</td><td> 500</td><td>insoluble</td>

Table B

<td>Weight ratio GADM -IPBC</td><td>Quantity in ppm</td><td>Solubility</td>

<td> 2000:1</td><td> 5</td><td>soluble</td>

<td> 1000:1</td><td> 10</td><td>soluble</td>

<td> 500:1</td><td> 20</td><td>soluble</td>

<td> 200:1</td><td> 50</td><td>soluble</td>

<td> 100:1</td><td> 100</td><td>soluble</td>

<td> 50:1</td><td> 200</td><td>insoluble</td>

<td> 20 :1</td><td> 500</td><td>insoluble</td>

Table C

<td>Weight ratio DMDMH IPBC</td><td>Quantity in ppm</td><td>Solubility</td>

<td> 2000:1</td><td> 5</td><td>soluble</td>

<td> 10001</td><td> 10</td><td>soluble</td>

<td> 500:1</td><td> 20</td><td>soluble</td>

<td> 2001</td><td> 50</td><td>soluble</td>

<td> 100:1</td><td> 100</td><td>soluble</td>

<td> 50:1</td><td> 200</td><td>insoluble</td>

<td> 20:1</td><td> 500</td><td>insoluble</td>

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These results show that compositions that are 50: 1 or 20: 1 mixtures are insoluble in water, while compositions that are 100: 1 to 2000: 1 mixtures are water-soluble.

synergism

Tables 1 to 14 show a very effective synergistic interaction between compounds (a) and (b) against a wide spectrum of Gram-negative and Gram-positive fungi and bacteria. The following microorganisms have been tested:

<td>microbe</td><td>ATCC number</td><td>Mokulum concentration</td>

<td>Ps aeruginosa (PSA) *</td><td> 9027</td><td>2.1 x 106 CFU / g product</td>

<td>E coli (ECOLI) *</td><td> 8739</td><td>4.7 x 10! CFU / g product</td>

<td>Staph aureus (SA) **</td><td> 6538</td><td>1.6 x 106 CFU / g product</td>

<td>Ps cepacia (PC) *</td><td> 25416</td><td>1.6 x 106 CFU / g product</td>

<td>C albicans (CAN) ***</td><td> 10231</td><td>8.0 x 104 CFU / g product</td>

<td>A niger (AN) ***</td><td> 16404</td><td>2.7 x 105 CFU / g product</td>

* / Gram-negative bacteria ** / Gram-positive bacteria *** / Fungi

Table D below shows static activity (MIC) and lethal activity of several antibacterial agents in ppm units. These activities were used to calculate the synergism index (SI) of the mixtures of the present invention.

Table D

Static activity (MIC) and lethal activity of a number of antibacterial compounds (static / lethal concentration ratio in ppm)

<td>microbe</td><td>ATCC number</td><td>IPBC</td><td>Germall® II</td><td>GADM</td><td>DMDMH</td><td>IPGA</td>

<td>ECOIL</td><td> 8739</td><td> 50/100</td><td> 400/1600</td><td> 400/800</td><td> 400/800</td><td> 150/600</td>

The synergism index was determined using the same mathematical transformations of this type of data as described by Kull et al in Applied Microbiology, 9, (1961), pages 538-541, using the following relationships:

O. Qr

Synergy index (SI) = - + - (2a (2 b where:

Qand = the amount of single acting compound "a" having the final effect. Qb = the amount of single acting "b" producing the final effect. Qand = the amount of "A" in the mixture producing the final effect.

Qb = the amount of "B" in the mixture producing the final effect.

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When SI equals 1, it means only an additional effect of the ingredients in the mixture, when SI is less than 1 it means synergism, whereas when SI is greater than 1, it means antagonism of two components.

According to these well-known methods of measuring synergism, the amount of each ingredient in different mixtures is compared with the amount of pure ingredient that is needed to achieve the same end result or to have the same microbial effect as the mixture.

Table 1

GERMALL® II / IPBC weight ratio - 2000 1

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,01%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 0,05</td><td> 99,95</td><td> 0,06</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,025%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 0,125</td><td> 249,9</td><td> 0,16</td>

<td>Level application</td><td>microbe</td><td>Q.</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,05%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 0,25</td><td> 499,8</td><td> 0,31</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,10%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 0,5</td><td> 999,5</td><td> 0,63</td>

181 504 cd. table 1

<td>Level of application</td><td>microbe</td><td>Q.</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,20%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 1</td><td> 1999</td><td> 1,26</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,40%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 2</td><td> 3998</td><td> 2,52</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,50%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 2,5</td><td> 4997,5</td><td> 3,15</td>

Table 2

GERMALL® II / IPBC weight ratio - 1000: 1

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,01%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 0,1</td><td> 99,9</td><td> 0,06</td>

<td>Level of application</td><td>microbe</td><td>Q.</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,025%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 0,25</td><td> 249,8</td><td> 0,16</td>

181 504 cd. table 2

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,05%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 0,5</td><td> 499,5</td><td> 0,32</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,10%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 1</td><td> 999</td><td> 0,63</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,20%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 2</td><td> 1998</td><td> 1,27</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,40%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 4</td><td> 3996</td><td> 2,54</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,50%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 5</td><td> 4995</td><td> 3,17</td>

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Table 3

GERMALL® II / IPBC weight ratio - 500.1

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,01%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 0,2</td><td> 99,8</td><td> 0,06</td>

<td>Level application</td><td>microbe</td><td>Q.</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,025%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 0,5</td><td> 249,5</td><td> 0,16</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,05%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 1</td><td> 499</td><td> 0,32</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,10%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 2</td><td> 998</td><td> 0,64</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,20%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 4</td><td> 1996</td><td> 1,29</td>

181 504 cd table 3

<td>Level of application</td><td>microbe</td><td>Q.</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,40%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 8</td><td> 3992</td><td> 2,58</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,50%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 10</td><td> 4990</td><td> 3,22</td>

Table 4

GERMALL® II / IPBC weight ratio - 200: 1

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,01%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 0,5</td><td> 99,5</td><td> 0,07</td>

<td>Level of application</td><td>microbe</td><td>Q.</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,025%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 1,25</td><td> 248,75</td><td> 0,17</td>

<td>Level of application</td><td>microbe</td><td>Q.</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,05%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 2,5</td><td> 497,5</td><td> 0,34</td>

181 504 cd. table 4

<td>Level of application</td><td>microbe</td><td>Q.</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,10%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 5</td><td> 995</td><td> 0,67</td>

<td>Level of application</td><td>microbe</td><td>Q.</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,20%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 10</td><td> 1990</td><td> 1,34</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,40%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 20</td><td> 3980</td><td> 2,69</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,50%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 25</td><td> 4975</td><td> 3,36</td>

Table 5

GERMALL® II / IPBC weight ratio - 100.1

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,01%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 1</td><td> 99</td><td> 0,07</td>

181 504 cd. table 5

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,025%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 2,5</td><td> 248</td><td> 0,18</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>Qb</td><td>SI</td>

<td> 0,05%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 5</td><td> 495</td><td> 0,36</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,10%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 10</td><td> 990</td><td> 0,72</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,20%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 20</td><td> 1980</td><td> 1,44</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,40%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 40</td><td> 3960</td><td> 2,88</td>

181 504 cd table 5

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,50%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 50</td><td> 4950</td><td> 3,59</td>

Table 6

GERMALL® II / IPBC weight ratio - 50 · 1

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,01%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 2</td><td> 98</td><td> 0,08</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,025%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 5</td><td> 245</td><td> 0,20</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,05%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 10</td><td> 490</td><td> 0,41</td>

<td>Level of application</td><td>microbe</td><td>Q.</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,10%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 20</td><td> 980</td><td> 0,81</td>

181 504 cd. table 6

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,20%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 40</td><td> 1960</td><td> 1,63</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,40%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 80</td><td> 3920</td><td> 3,25</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,50%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 100</td><td> 4900</td><td> 4,06</td>

Table 7

GERMALL® II / IPBC weight ratio - 20 1

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,01%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 5</td><td> 95</td><td> 0,11</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,025%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 12,5</td><td> 237,5</td><td> 0,27</td>

181 504 cd table 7

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,05%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 25</td><td> 475</td><td> 0,55</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,10%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 50</td><td> 950</td><td> 1,09</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,20%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 100</td><td> 1900</td><td> 2,19</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,40%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 200</td><td> 4800</td><td> 5,00</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,50%</td><td>ecoli</td><td> 100</td><td> 1600</td><td> 250</td><td> 4750</td><td> 5,47</td>

181 504

Table 8

DMDMH / IPBC weight ratio - 2000 1

<td>Level application</td><td>microbe</td><td>Q.</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,01%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 0,05</td><td> 99,95</td><td> 0,13</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,025%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 0,125</td><td> 249,88</td><td> 0,31</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,05%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 0,25</td><td> 499,75</td><td> 0,63</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,10%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 0,5</td><td> 999,5</td><td> 1,25</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,20%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 1</td><td> 1999</td><td> 2,51</td>

181 504 cd. table 8

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,40%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 2</td><td> 3998</td><td> 5,02</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,50%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 2,5</td><td> 4997,5</td><td> 6,27</td>

Table 9

DMDMH / IPBC weight ratio - 1000 '1

<td>Level of application</td><td>microbe</td><td>Q.</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,01%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 0,1</td><td> 99,9</td><td> 0,13</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,025%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 0,25</td><td> 249,8</td><td> 0,31</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,05%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 0,5</td><td> 499,5</td><td> 0,63</td>

181 504 cd table 9

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,10%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 1</td><td> 999</td><td> 1,26</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,20%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 2</td><td> 1998</td><td> 2,52</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,40%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 4</td><td> 3996</td><td> 5,04</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,50%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 5</td><td> 4995</td><td> 6,29</td>

Table 10

DMDMH / IPBC weight ratio - 500.1

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,01%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 0,2</td><td> 99,8</td><td> 0,13</td>

181 504 cd. table 10

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,025%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 0,5</td><td> 249,5</td><td> 0,32</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,05%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 1</td><td> 499</td><td> 0,63</td>

<td>Level application</td><td>microbe</td><td>Q.</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,10%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 2</td><td> 998</td><td> 1,27</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,20%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 4</td><td> 1996</td><td> 2,54</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qe</td><td>SI</td>

<td> 0,40%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 8</td><td> 3992</td><td> 5,07</td>

181 504 cd. table 10

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,50%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 10</td><td> 4900</td><td> 6,23</td>

Table 11

DMDMH / IPBC weight ratio. - 200: 1

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,01%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 0,5</td><td> 99,5</td><td> 0,13</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,025%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 1,25</td><td> 248,75</td><td> 0,32</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,05%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 2,5</td><td> 497,5</td><td> 0,65</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,10%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 5</td><td> 995</td><td> 1,29</td>

181 504 cd of table 11

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,20%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 10</td><td> 1990</td><td> 2,59</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,40%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 20</td><td> 3980</td><td> 5,18</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,50%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 25</td><td> 4975</td><td> 6,47</td>

Table 12

DMDMH / IPBC weight ratio - 100.1

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,01%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 1</td><td> 99</td><td> 0,13</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,025%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 2,5</td><td> 248</td><td> 0,33</td>

181 504 cd table 12

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,05%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 5</td><td> 495</td><td> 0,67</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,10%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 10</td><td> 990</td><td> 1,34</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,20%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 20</td><td> 1980</td><td> 2,68</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,40%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 40</td><td> 3960</td><td> 5,35</td>

<td>Level application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,50%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 50</td><td> 4950</td><td> 6,69</td>

181 504

Table 13

DMDMH / IPBC weight ratio - 50 · 1

<td>Level application</td><td>microbe</td><td>Q.</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,01%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 2</td><td> 98</td><td> 0,14</td>

<td>Level of application</td><td>microbe</td><td>Q.</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,025%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 5</td><td> 245</td><td> 0,36</td>

<td>Level of application</td><td>microbe</td><td>Q.</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,05%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 10</td><td> 490</td><td> 0,71</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,10%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 20</td><td> 980</td><td> 1,43</td>

<td>Level of application</td><td>microbe</td><td>Q.</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,20%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 40</td><td> 1960</td><td> 2,85</td>

181 504 cd. table 13

<td>Level of application</td><td>microbe</td><td>Q.</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,40%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 80</td><td> 3920</td><td> 5,70</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,50%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 100</td><td> 4900</td><td> 7,13</td>

Table 14

DMDMH / IPBC weight ratio - 20 Ί

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,01%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 5</td><td> 95</td><td> 0,17</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>Ob</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,025%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 12,5</td><td> 237,5</td><td> 0,42</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,05%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 25</td><td> 475</td><td> 0,84</td>

181 504 cd. table 14

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,10%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 50</td><td> 950</td><td> 1,69</td>

<td>Level of application</td><td>microbe</td><td>Q.</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,20%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 100</td><td> 1900</td><td> 3,38</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,40%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 200</td><td> 4800</td><td> 8,00</td>

<td>Level of application</td><td>microbe</td><td>qa</td><td>qb</td><td>qa</td><td>qb</td><td>SI</td>

<td> 0,50%</td><td>ecoli</td><td> 100</td><td> 800</td><td> 250</td><td> 4750</td><td> 8,44</td>

Similar SI values were also determined using the hydroxymethyl group GADM and SUTTOCIDE® A in place of GERMALL® II or DMDMH, in a mixture with IPBC, in the same weight ratios and ranges of levels of use, as shown in Tables 1-14 above.

Tables 1-14 above illustrate the synergism of IPBC (compound B) in combination with GERMALL® II or DMDMH (compound A), in A: B weight ratios of 2000: 1, 1000.1,500: 1,200: 1,100: 1,50: 1 , and 20: 1. The synergistic effect is very effective for all proportions used, at low levels of use, for example from 0.01% to 0.1%, against all gram-positive and gram-negative microorganisms tested and fungi. At slightly higher levels of use, for example from 0.20% to 0.50%, the compounds in all tested proportions acted synergistically against Candida albicans as well as A.niger. However, at proportions ranging from 50: 1 to 20: 1 and

181 504 at levels of use from 0.01% to 0.1%, the synergistic effect is insignificant, as well as no synergistic effect was found for Candida albicans at levels of use from 0.20% to 0.50%.

In the case of GERMALL® II in the composition as a hydroxymethyl containing compound, the SI values were lower compared to DMDMH.

Similar results were obtained when iodopropynyl alcohol (IPGA) was used in place of IPBC in the compositions described above.

5. Protective activity (provocation test)

To perform the microbiological challenge test, a typical cosmetic emulsion was prepared and the previously prepared compositions of the hydroxymethyl group containing compound and IPBC were added in amounts corresponding to different levels of use. The emulsion thus produced has the following composition:

A phase by weight

Stearic acid 5.00

Mineral oil 2.50

Cetyl alcohol 1.00

Lareth-5 + Ceteth-5 + Oleth-5 + Steareth-5 0.5 ^ 0

Glycerin monostearyman + polyoxyethylene glycol stearate 1, 55)

Phase B

Demineralized water 88.0

Triethanolamine 99% 1.00

Citric acid 30% aqueous solution 0.60

Protective mixture qs

To form the emulsion, phases A and B were heated separately to 75 ° -80 ° C. Then, phase A was added to phase B with stirring. Then the mixture was cooled to 00 ° -00 ° C. At this point, the desired amount of protective composition was added and the whole was cooled to 50 ° C with stirring. Then, to adjust the pH, a citric acid solution was added and stirred until the temperature reached 30 ° C.

The following microorganisms were used for challenge tests: SA, ECOLI, PSA, PC, AN and CAN, and were performed as follows. Measured portions of the tested emulsion in an amount of 50 g, containing various amounts of the protective composition, were inoculated with the microorganisms used for the challenge in the amount of7-108. Then, the tested samples were mixed to disperse the provocative inoculum into them. Samples were incubated and analyzed after 48 hours, 7,14,211 28 days. The determinations were made using 1 g of test sample, serially diluting the initial concentration by 101 to 106 times. The medium for bacteria was Letheen agar, while for fungi the medium was Mycophil agar with low pH together with Tween 20. Each sample on the plate was incubated on bacteria for 48 hours at 37 ° C, on mold for 5 days at 25 ° C and for fungi for 3 days at 25 ° C. After incubation, the number of colonies per milliliter (cfu / ml) was calculated. After 21 days, the test product was re-inoculated with half of the initial inoculum. The results are shown below in Tables 15-23.

Table 15

GERMALL® II / IPBC in a ratio of 2000 · 1

<td>Level of testing</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,01%</td><td>ecoli</td><td> 110000</td><td> 290000</td><td> 2400</td><td> 138000</td><td> 560000</td>

181 504 cd. table 15

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,025%</td><td>ecoli</td><td> 39000</td><td> 10</td><td> <10</td><td> <10</td><td> <10</td>

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,05%</td><td>ecoli</td><td> 3400</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,1%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,2%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

Control test, mechromon

<td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td>ecoli</td><td> 54000</td><td> 140000</td><td> 170000</td><td> 170000</td><td> 74000</td>

181 504

Inoculum concentration

<td>microbe</td><td>0 hours</td><td>21 days</td>

<td>ecoli</td><td> 3600000</td><td> 170000</td>

Table 16

GERMALL® II / IPBC in a ratio of 1000.1

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,01%</td><td>ecoli</td><td> 120000</td><td> 63000</td><td> 93000</td><td> 92000</td><td> 9600000</td>

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 dm</td>

<td> 0,025%</td><td>ecoli</td><td> 120000</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,05%</td><td>ecoli</td><td> 68000</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,1%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

181 504 cd of table 16

<td>Level of testing</td><td>microbe</td><td>48 hours</td><td>7 dm</td><td>14 dm</td><td>21 dm</td><td>28 days</td>

<td> 0,2%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

Control test, unprotected

<td>microbe</td><td>48 hours</td><td>7 dm</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td>ecoli</td><td> 54000</td><td> 140000</td><td> 170000</td><td> 170000</td><td> 74000</td>

Lnokulum concentration

<td>microbe</td><td>0 hours</td><td>21 days</td>

<td>ecoli</td><td> 3600000</td><td> 170000</td>

Table 17

GERMALL® Π / IPBC in a ratio of 500.1

<td>Level of testing</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 dm</td><td>28 dm</td>

<td> 0,01%</td><td>ecoli</td><td> 90000</td><td> 57000</td><td> 95000</td><td> 70000</td><td> 240000</td>

<td>Level of testing</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 dm</td>

<td> 0,05%</td><td>ecoli</td><td> 60000</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

181 504 cd of table 17

<td>Level of testing</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,1%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

<td>Level tested</td><td>microbe</td><td>48 hours</td><td>7 dm</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,2%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

Control test, unprotected

<td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 dm</td>

<td>ecoli</td><td> 7100000</td><td> 6200000</td><td> 610000</td><td> 350000</td><td> 120000</td>

Inoculum concentration

<td>microbe</td><td>0 hours</td><td>21 dm</td>

<td>ecoli</td><td> 3900000</td><td> 1480000</td>

Table 18

GERMALL® II / IPBC in a ratio of 200.1

<td>Level of testing</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 dm</td>

<td> 0,01%</td><td>ecoli</td><td> 220000</td><td> 7600</td><td> <10</td><td> 850</td><td> >1000000</td>

181 504 cd. table 18

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,05%</td><td>ecoli</td><td> 37000</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,1%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,2%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

Control test, unprotected

<td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td>ecoli</td><td> 640000</td><td> 360000</td><td> 410000</td><td> 990000</td><td> 68000</td>

Inoculum concentration

<td>microbe</td><td>0 hours</td><td>21 days</td>

<td>ecoli</td><td> 5800000</td><td> 1300000</td>

181 504

Table 19

GERMALL® II / IPBC in a 50 · 1 ratio

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,01%</td><td>ecoli</td><td> 170000</td><td> 710</td><td> 10</td><td> <10</td><td> 120</td>

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,05%</td><td>ecoli</td><td> 150000</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,1%</td><td>ecoli</td><td> 510</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,2%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

Control test, unprotected

<td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td>ecoli</td><td> 640000</td><td> 360000</td><td> 410000</td><td> 990000</td><td> 68000</td>

181 504

Inoculum concentration

<td>microbe</td><td>0 hours</td><td>21 days</td>

<td>ecoli</td><td> 5800000</td><td> 1300000</td>

Table 20

GERMALL® II / IPBC in a ratio of 20 1

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,01%</td><td>ecoli</td><td> 160000</td><td> 110</td><td> <10</td><td> <10</td><td> 20</td>

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,05%</td><td>ecoli</td><td> 16000</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,1%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,2%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

181 504

Control test, unprotected

<td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td>ecoli</td><td> 640000</td><td> 360000</td><td> 410000</td><td> 990000</td><td> 68000</td>

Inoculum concentration

<td>microbe</td><td>0 hours</td><td>21 days</td>

<td>ecoli</td><td> 5800000</td><td> 1300000</td>

Table 21

DMDMH / IPBC in a ratio of 2000 '1

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,025%</td><td>ecoli</td><td> 1300000</td><td> <10</td><td> <10</td><td> <10</td><td> 3000</td>

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,05%</td><td>ecoli</td><td> 220000</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 dm</td><td>28 days</td>

<td> 0,1%</td><td>ecoli</td><td> 340</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

181 504 cd. Table 21

<td>Level of testing</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,2%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

Control test, unprotected

<td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td>ecoli</td><td> 150000</td><td> 2500000</td><td> 7300000</td><td> 240000</td><td> 140000</td>

Inoculum concentration

<td>microbe</td><td>0 hours</td><td>21 days</td>

<td>ecoli</td><td> 4800000</td><td> 5800000</td>

Table 22

DMDMH / IPBC in a ratio of 1000: 1

<td>Level of testing</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,025%</td><td>ecoli</td><td> 160000</td><td> <10</td><td> <10</td><td> <10</td><td> 2400</td>

<td>Level of testing</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,05%</td><td>ecoli</td><td> 26000</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

181 504 cd of table 22

<td>Level of testing</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,1%</td><td>ecoli</td><td> 45000</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

<td>Level of testing</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,2%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

Control test, unprotected

<td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td>ecoli</td><td> 150000</td><td> 2500000</td><td> 7300000</td><td> 240000</td><td> 140000</td>

Inoculum concentration

<td>microbe</td><td>0 hours</td><td>21 days</td>

<td>ecoli</td><td> 4800000</td><td> 5800000</td>

Table 23

GADM / IPBC in a ratio of 2000: 1

<td>Level of testing</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,025%</td><td>ecoli</td><td> 230000</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

181 504 cd of table 23

<td>Level of testing</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,05%</td><td>ecoli</td><td> 120000</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

<td>Level of testing</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,1%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

<td>Level of testing</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,2%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

Control test, unprotected

<td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td>ecoli</td><td> 600000</td><td> 3100000</td><td> 920000</td><td> 920000</td><td> 140000</td>

Inoculum concentration

<td>microbe</td><td>0 hours</td><td>21 days</td>

<td>ecoli</td><td> 170000</td><td> 3500000</td>

Discussion of the results of the provocation test

The results of the 28-day challenge test presented above in Tables 15-23 show the effectiveness of the protective mixture according to the invention against a broad spectrum of bacteria and fungi in the usable emulsion formulation.

181 504

For example, admixtures in the form of GERMALL® II and IPBC compositions in a weight ratio of 2000: 1 (Table 15) when their level of use is from 0.05% to 0.2%, which corresponds to a content from 0.75 ppm to 10 ppm IPBC and from 500 ppm to 2000 ppm of a hydroxymethyl containing compound, in principle, provide complete protection after 28 days against all tested microorganisms. With a low level of use of the active ingredient of 0.05%, all challenge tests were carried out within 21 days. Then, after re-vaccination after 21 days, all microorganisms were killed within 7 days, except for CAN, which were lysed within 14 days.

Table 21 shows the results of the challenge test for mixtures of DMDMH and IPBC, with the same weight ratio of 2000: 1. However, it is necessary to use this mixture at an application level of 0.2% in order to act against all microorganisms after 21 days. After re-vaccination, all germs were killed within 7 days, except for CAN which were lysed within 14 days. Based on these results, it is clear that GERMALL® II mixed with IPBC is four times more effective than the DMDMH / IPBC mixture.

GERMALL® II, as shown in Table 23, is also better than GADM as a hydroxymethyl containing compound.

Table 24 below provides the results of a study on the activity of the GII / IPBC solution in propylene glycol. Compositions of active ingredients GII and IPBC were prepared in weight ratios of 99.5% / 0.5% and 99% / 1% and added to 60% by weight of propylene glycol. The resulting solutions were tested at a total active ingredient content in proteinaceous shampoo of 0.05%, 0.1% and 0.2%.

Table 24

99.5% GII / 0.5% IPBC solution in propylene glycol - total active ingredient - 0.05%

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 dm</td><td>28 dm</td>

<td> 0,05%</td><td>ecoli</td><td> 3000</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

99.5% GII / 0.5% IPBC solution in propylene glycol - total active ingredient - 0.1%

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,1%</td><td>ecoli</td><td> 580</td><td> <10</td><td> <10</td><td> <10</td><td> 20</td>

181 504 cd. table 24

99.5% GII / 0.5% IPBC solution in propylene glycol - total active ingredient - 0.2%

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,2%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> 20</td>

99% GII / 1% IPBC solution in propylene glycol - total active ingredient - 0.05%

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,05%</td><td>ecoli</td><td> 3300</td><td> <10</td><td> <10</td><td> <10</td><td> 60</td>

99% GII / 1% IPBC solution in propylene glycol - total active ingredient - 0.1%

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,1%</td><td>ecoli</td><td> 840</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

99% GII / 1% IPBC solution in propylene glycol - total active ingredient - 0.2%

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,2%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

181 504

Control check, unprotected

<td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td>ecoli</td><td></td><td> >100000000</td><td> 800000000</td><td> 48000000</td><td> 600000</td>

Inoculum concentration

<td>microbe</td><td>0 hours</td><td>21 days</td>

<td>ecoli</td><td> 2500000</td><td> 5300000</td>

Table 25 presents the test results similar to those shown above in Table 24, in which the carrier for the composition was the previously described typical emulsion.

Tables 1 and 25

99.5% GII / 0.5% IPBC solution in propylene glycol - total active ingredient - 0.05%

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,05%</td><td>ecoli</td><td> 320</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

99.5% GII / 0.5% IPBC solution in propylene glycol - total active ingredient - 0.1%

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 din</td>

<td> 0,1%</td><td>ecoli</td><td> 10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

181 504 cd of table 25

99.5% GII / 0.5% IPBC solution in propylene glycol - total active ingredient - 0.2%

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,2%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

99% GII / 1% IPBC solution in propylene glycol - total active ingredient - 0.05%

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 dm</td><td>28 days</td>

<td> 0,05%</td><td>ecoli</td><td> 66000</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

99% GII / 1% IPBC solution in propylene glycol - total active ingredient - 0.1%

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,1%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

99% GII / 1% IPBC solution in propylene glycol - total active ingredient - 0.2%

<td>Level test</td><td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td> 0,2%</td><td>ecoli</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td><td> <10</td>

181 504

Control test, non-rotating

<td>microbe</td><td>48 hours</td><td>7 days</td><td>14 days</td><td>21 days</td><td>28 days</td>

<td>ecoli</td><td> 11000000</td><td> 7300000</td><td> 6000000</td><td> 730000</td><td> 220000</td>

Inoculum concentration

<td>microbe</td><td>0 hours</td><td>21 days</td>

<td>ecoli</td><td> 2500000</td><td> 5300000</td>

The results in Tables 24 and 25 show that the compositions of the invention are fully effective against the microorganisms tested, compared to non-selective controls.

Although the invention has been described with particular representation of certain variants thereof, it should be understood that changes and modifications may be made which fall within its scope.

181 504

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Priority claims8

Priority claims
Document	Office	Kind	Date
23408994	United States of America	A	1994-04-28
41374295	United States of America	A	1995-03-30
9504895	United States of America	W	1995-04-21
234089	–	–	–
US19940234089	–	–	–
US19950413742	–	–	–
US9504895	–	–	–
WO1995US04895	–	–	–

Members23

Family members, oldest first
Document	Office	Kind
US5428050A	United States of America	A	1995-06-27
CA2183652A1	Canada	A1	1995-11-09
WO9529588A1	World Intellectual Property Organization (WIPO)	A1	1995-11-09
AU2392395A	Australia	A	1995-11-29
US5496842A	United States of America	A	1996-03-05
US5552425A	United States of America	A	1996-09-03
HU9602963D0	Hungary	D0	1996-12-30
NZ284708A	New Zealand	A	1997-01-29
EP0757518A1	European Patent Office (EPO)	A1	1997-02-12
PL316963A1	Poland	A1	1997-02-17
KR970701496A	Republic of Korea	A	1997-04-12
US5631273A	United States of America	A	1997-05-20
HUT76978A	Hungary	A	1998-01-28
EP0757518A4	European Patent Office (EPO)	A4	1998-02-25
AU691613B2	Australia	B2	1998-05-21
EP0757518B1	European Patent Office (EPO)	B1	2001-05-30
DE69521128D1	Germany	D1	2001-07-05
PL181504B1This record	Poland	B1	2001-08-31
ES2158106T3	Spain	T3	2001-09-01
DE69521128T2	Germany	T2	2001-11-08
KR100377974B1	Republic of Korea	B1	2003-06-12
CA2183652C	Canada	C	2007-09-04
HU228148B1	Hungary	B1	2012-12-28

Water-soluble synergistic protective compositions of lethal compounds

Abstract

Term

Priority claims8

Members23

Events

Numbers

Titles

Classification