Alcoholic disinfectant with good skincare effect and high efficacy

The present invention relates to an alcoholic disinfectant, and to the use of the disinfectant for the disinfection of surfaces, in particular for hygienic and surgical hand disinfection. Furthermore, the invention relates to the use of fatty alcohol for improving the antimicrobial efficacy of an alcoholic disinfectant.

When using alcoholic hand disinfectants, the refatting of the skin plays a major role for user compliance. Glycerol is described in the literature as having skincare properties (Atrux-Tallau, N., Romagny, C, Padois, K., Denis, A., Haftek, M., Falson, F., Pirot, F., and Maibach, H.I. (2010), Arch Dermatol Res 302:435 - 441 ). Glycerol is therefore a widely used substance as refatting agent in commercial formulations for alcoholic hand disinfection.

WO 2014/032696 A1 discloses virucidal alcoholic hand disinfectants. Besides C _2- to C ₃ -alcohol, the presence of glycerol is mandatory. However, investigations with glycerol-containing formulations (WHO-I and WHO-II) have shown that the presence of glycerol inhibits the bactericidal properties of alcoholic hand disinfectants for surgical hand disinfection according to EN12791 (Suchomel, M., Rotter, M., Weinlich, M., Kundi, M. (2013). J. Hosp Infect 83:284- 7).

DE 10 2014 1 15 080 A1 describes ethanolic disinfectants which comprise a) at least 78% by weight of ethanol and b) one or more fatty acid esters of monovalent d- to C ₆ -alkyl alcohol. The addition of the fatty acid ester as refatting agent leads to an improvement in the efficacy in the quantitative suspension test according to DGHM.

WO 2009/088894 A1 discloses compositions which comprise at least 35% by weight of C _2- to C ₅ -alcohol, hydrophobic polymer soluble in the alcohol, cationic antimicrobial agent and skincare ester. It is also described that fatty alcohols increase the bactericidal efficacy of the cationic antimicrobial agents.

DE 10 2010 054 155 A1 discloses the use of aliphatic alcohols in the production of a low-spore or spore-free disinfectant for preventing contamination of the disinfectant with spores.

WO 2014/100 851 A1 describes alcoholic compositions with a content of at least 0.2% by weight of isopropyl myristate. US 2003/0139307 A1 discloses anhydrous skin cleaning compositions with a content of alcohol and silicone- based materials.

US 2012/0208894 A1 describes ethanolic antiseptics with a content of cyclomethicones or isohexadecane as skincare components. DE 10 2005 002 645 A1 describes alcoholic compositions with a content of glycerol ether and bispyridinium alkane.

DE 10 2013 213 252 A1 discloses an isopropanol-containing liquid composition for use in a method for controlling caliciviruses. The presence of fatty alcohol as refatting agent improves the efficacy against caliciviruses. Furthermore, a disinfectant with a content of 75% by weight of isopropanol and efficacy in hygienic hand disinfection (EN1500) and surgical hand disinfection (EN12791 ) is known which comprises a combination of dexpanthenol and 1 -(2-ethylhexyl) glycerol ether as skin protection and skincare.

EP 2 135 507 A1 discloses ethanolic disinfectants with a content of a compound which releases zinc ions in solution. EP 1 685 854 A1 discloses virucidal disinfectants with a content of acidic phosphorous compounds.

The present invention is based on the object of providing refatting agents for disinfectants with a content of isopropanol and ethanol, where the refatting agents in the disinfectant should have, in contrast to glycerol, no effect inhibiting the antimicrobial efficacy, and indeed in particular if they are to be used in a relatively high concentration.

There is, in particular, a need for disinfectants for hygienic and surgical hand disinfection which despite having a high content of C ₂ - to C ₃ -alcohols and accordingly being very highly antimicrobially (in particular bactericidally) effective, at the same time have the least possible defatting effect. In this connection, these agents should comprise the fewest possible mandatory constituents. The disinfectants should thus not comprise many different mandatory antimicrobial active ingredients. Similarly, the agents should not automatically comprise many different active ingredients for improving the skincare properties (or these skincare active ingredients in a large amount).

It has now been found that this object is achieved according to the invention through the use of fatty alcohols. According to the present invention, it has thus been found that the addition of fatty alcohols as refatting agents to a disinfectant with a content of isopropanol and ethanol even leads to an improvement in the efficacy towards bacteria both in the quantitative suspension experiment according to EN 13727 as well as in the germ carrier experiment simulating practical conditions in accordance with EN 12791 , where, surprisingly, it does not depend on the presence of glycerol or glycerol ethers (such as 1 -(2- ethylhexyl) glycerol ethers). Surprisingly, it has been possible to show that the use according to the invention of fatty alcohols, in combination with the effective constituents isopropanol and ethanol, leads to a significant increase in efficacy. This is surprising in particular against the background that the fatty alcohols without 1 -(2- ethylhexyl) glycerol ethers lead to a considerably improved efficacy of the formulation against bacterial pathogens, which differ considerably from viral pathogens in their cellular structure. In this connection, favourable effects have been shown both against GRAM-negative bacteria (£. coli) as well as against G RAM-positive pathogens on the skin (see the experiments relating to surgical hand disinfection).

In a first aspect, the invention thus relates to a

, which comprises for 100% of its weight:

a) 50 to 85% by weight of isopropanol,

b) 3 to 20% by weight of ethanol and

c) one or more fatty alcohols,

said composition also comprising less than 0.3% by weight of glycerol. a) Isopropanol

The composition according to the invention comprises 50 to 85% by weight of isopropanol. It is preferred that the content of a) isopropanol is 55 to 83% by weight, preferably 60 to 80% by weight, more preferably 65 to 75% by weight, such as 68 to 72% by weight.

b) Ethanol

The composition according to the invention comprises 3 to 20% by weight of ethanol. It is preferred that the content of b) ethanol is 4 to 18% by weight, preferably 6 to 14% by weight, more preferably 8 to 12% by weight, such as 9 to 1 1 % by weight.

c) Fatty alcohols

The composition according to the invention comprises one or more fatty alcohols. It is preferred that the content of c) fatty alcohol is 0.2 to 2% by weight, preferably 0.4 to 1.5% by weight, more preferably 0.5 to 1 .2% by weight, in particular 0.6 to 1 .0% by weight, such as 0.7 to 0.9% by weight.

The fatty alcohols used according to the invention are monohydric, primary alcohols and are derived from unbranched, optionally mono- or polyunsaturated hydrocarbon radicals having 6 to 22 carbon atoms, more particularly 12 to 18 carbon atoms. Examples of fatty alcohols are lauryl alcohol, myristyl alcohol, cetyl alcohol and stearyl alcohol. In a preferred embodiment, component c) consists of a mixture of two or more fatty alcohols. Preferably, component c) in the disinfectant consists of a combination of three or more fatty alcohols, such as, for example, the mixture of cetylstearyl alcohol (cetearyl alcohol, i.e. the mixture of cetyl alcohol and steary alcohol) with myristyl alcohol.

According to the invention, component c) is thus preferably a mixture consisting of i) cetyl alcohol, ii) stearyl alcohol and iii) myristyl alcohol.

According to a particular embodiment, the composition as herinbefore defined comprises for 100% of its weight:

a) 60 to 80% by weight of isopropanol,

b) 6 to 14% by weight of ethanol and

c) 0.4 to 1 .5% by weight of a mixture consisting of i) cetyl alcohol, ii) stearyl alcohol and iii) myristyl alcohol.

The disinfectant according to the invention comprises less than 0.3% by weight of glycerol. It is preferred that the disinfectant according to the invention comprises less than 0.25% by weight of glycerol, preferably less than 0.2% by weight, in particular less than 0.1 % by weight, such as less than 0.05% by weight or less than 0.01 % by weight, of glycerol. Particularly preferably, no glycerol is present in the disinfectant according to the invention.

In a preferred embodiment, the disinfectant according to the invention comprises less than 0.25% by weight of glycerol ethers, preferably less than 0.2% by weight of glycerol ethers, in particular less than 0.1 % by weight of glycerol ethers, such as less than 0.05% by weight or less than 0.01 % by weight of glycerol ethers. Particularly preferably, no glycerol ether is present in the disinfectant according to the invention.

In a further preferred embodiment, the disinfectant according to the invention comprises less than 0.25% by weight of 1 -(2-ethylhexyl) glycerol ether, preferably less than 0.2% by weight of 1 -(2-ethylhexyl) glycerol ether, in particular less than 0.1 % by weight of 1 -(2-ethylhexyl) glycerol ether, such as less than 0.05% by weight or less than 0.01 % by weight of 1 -(2-ethylhexyl) glycerol ether. Particularly preferably, no 1 -(2-ethylhexyl) glycerol ether is present in the disinfectant according to the invention.

In a further embodiment according to the invention, the disinfectant comprises less than 10% by weight of n-propanol, preferably less than 8% by weight of n-propanol, in particular less than 5% by weight of n-propanol, such as less than 2% by weight or less than 1 % by weight of n-propanol. Particularly preferably, no n-propanol is present in the disinfectant according to the invention. Further constituents of the disinfectant

Besides the mandatory components a), b) and c), the composition according to the invention can also comprise further components:

d) Sugar alcohol

The composition according to the invention can comprise:

d) one or more sugar alcohols, where the component d) is preferably sorbitol.

It is preferred that component d) is present in an amount of from 0.01 to 2% by weight, more preferably 0.1 to 2% by weight, in particular 0.2 to 1 .5% by weight, such as 0.3 to 1 .0% by weight, for example about 0.5% by weight.

According to another particular embodiment, the composition as herinbefore defined comprises for 100% of its weight:

a) 60 to 80% by weight of isopropanol,

b) 6 to 14% by weight of ethanol,

c) 0.4 to 1 .5% by weight of a mixture consisting of i) cetyl alcohol, ii) stearyl alcohol and iii) myristyl alcohol;

d) 0.1 to 2% by weight of sorbitol.

e) Skincare agents

The composition according to the invention can comprise e) skincare agents, where component e) is preferably urea, dexpanthenol, hamamelis, bisabolol and/or allantoin, where component e) is in particular dexpanthenol.

In a preferred embodiment, component e) (in particular dexpanthenol) is present preferably in an amount of from 0.01 to 1 % by weight, preferably 0.05 to 0.5% by weight, such as 0.1 to 0.3% by weight, for example about 0.2% by weight.

According to another particular embodiment, the composition as herinbefore defined comprises for 100% of its weight:

a) 60 to 80% by weight of isopropanol;

b) 6 to 14% by weight of ethanol;

c) 0.4 to 1 .5% by weight of a mixture consisting of i) cetyl alcohol, ii) stearyl alcohol and iii) myristyl alcohol;

d) 0.1 to 2% by weight of sorbitol; and

e) 0.05 to 0.5% by weight of dexpanthenol.

According to an usual embodiment, the composition as herinbefore defined also comprises water.

According to another particular embodiment, the composition as herinbefore defined consists of 100% of its weight: a) 65 to 75% by weight of isopropanol;

b) 8 to 12% by weight of ethanol;

c) 0.5 to 1 .2% by weight of a mixture consisting of i) cetyl alcohol, ii) stearyl alcohol and iii) myristyl alcohol;

d) 0.2 to 1 .5% by weight of sorbitol; and

e) 0.1 to 0.3% by weight of dexpanthenol, and

f) water q.s. 100% by weight.

Application

The composition according to the invention are preferably used for skin disinfection, preferably for the disinfection of human skin, and in particular for hand disinfection.

The invention also relates to the use of the composition as hereinbefore defined, to disinfect the surfaces, where the surface is preferably an animate surface. Preferably, the animate surface is skin, in particular human skin, where the use takes place in particular in the case of surgical hand disinfection (efficacy test according to EN12791 ) or hygienic hand disinfection (efficacy test according to EN 1500). That is why, the invention also relates to the use of the composition as hereinbefore defined, for use as a disinfectant of the human or animal body skin, and more particularly for use as a disinfectant of the human hand skin.

In a third aspect, the present invention relates to the use of fatty alcohol for improving the antimicrobial efficacy of an alcoholic disinfectant, where the disinfectant comprises a) 50 to 85% by weight of isopropanol and b) 3 to 20% by weight of ethanol, but less than 0.3% by weight of glycerol. Preferably, the improved antimicrobial efficacy is an improvement in the efficacy in the case of surgical hand disinfection according to EN 12791.

The invention also relates to a mMethod for the hand disinfection in a preoperative preparation of an operation, or of any medical treatment, comprising a step of rubbing the hands with the composition as hereinbefore defined.

The advantages of the present invention arise in particular from the following examples. Unless stated otherwise, all of the percentages refer to the weight.

EXAMPLES The following formulations were tested (Tab. 1 ). Tab. 1 . Tested formulations

^* Comparison

¹ Methyl ethyl ketone

METHODS AND EXPERIMENTAL PROCEDURE

Method 1 : Testing the suitability of formulation I for "Surgical hand disinfection" (DIN prEN12791)

The test was performed with 20 subjects; the experiment was conducted one week apart after the "cross-over arrangement" corresponding to DIN prEN12791 (phase 2 stage 2). In the reference procedure, 60% by volume of n-propanol was placed in portions into the hands for rubbing over 3 minutes. The test preparation was applied in the same manner over 3 minutes. The results are given in Tables 2a and 2b, and 2c and 2d and summarized in Tab. 5.

Table 2a. Surgical reference hand disinfection - experimental results 30.11.2000, experimental data: 12.10.2000

Preparation: reference preparation (propan-1 -ol with a volume concentration of 60%), application: rubbing in for 3 min.

Subject number CFU per plate from dilution 10 ^X Hand left or Before values After values right Immediate value

-1 -2 -3 0 -1 -2

I nc 220 21 1 15 19 1 nc 266 19

I nc 196 13 67 4 0 nc 137 12

I nc 135 12

nc 198 22 174 19 1

I nc 177 21

nc 150 20 101 9 0

I nc 195 26

nc 129 19 87 7 0

I nc 142 19 73 6 0 nc 180 23

I nc 147 18 98 7 0 nc 1 14 12

I nc 130 24 47 4 0 nc 187 19

I nc 277 19

nc 1 19 21 167 1Z 1

I nc 95 1 1 121 22 1 nc 65 8

I nc 180 21 30 1 0 nc 210 24

I nc nc 82 nc 57 6 nc nc 50

I nc 1 16 9 174 13 1 nc 145 7

I nc nc 48 nc 63 18

nc nc 63

I nc 180 22 97 8 0 nc 215 26

I nc 133 18

r nc 187 32 54 4 0 17 I nc 126 16 68 5 0 r nc 173 19

18 I nc 212 39

r nc 209 26 nc 47 5

19 I nc 283 37 180 13 1

r nc 219 24

20 I nc 223 20

r nc 228 25 Z4 6 0 underlined number used for further calculations

use weighted average

nc not countable Table 2b. List of calculated log values and log reduction factors 30.11.2000

Preparation: reference preparation (propan-1-ol with a volume concentration of 60%)

Application: rubbing in for 3 min.

Subject Immediate effect

No. log x logy log z

1 4.34 2.09 2.25

2 4.29 1.83 2.46

3 4.30 2.24 2.06

4 4.19 2.00 2.19

5 4.13 1.94 2.19

6 4.16 1.86 2.30

7 4.18 1.99 2.19

8 4.15 1.67 2.48

9 4.10 2.22 1.88

10 3.98 2.11 1.87

11 4.26 1.48 2.78

12 4.91 2.76 2.15

13 4.06 2.24 1.82

14 4.68 2.87 1.81

15 4.26 1.99 2.27

16 4.30 1.73 2.57

17 4.11 1.83 2.28

18 4.33 2.67 1.66 19 4.46 2.26 2.20

20 4.36 1 .87 2.49

X 4.28 2.08 2.20

s 0.15 0.38 0.24

N 20 20 20

log y = log after value

s = standard deviation

log z = log-reduction factor

N = number of values (subjects) in each column

Table 2c. Surgical hand disinfection with the product to be tested - experimental results -

Preparation: formulation I

Experiment date: 21.10.2000

Application: rubbing in for 3 min.

Subject number CFU per plate from dilution 10 ^X

No. Hand left or Before values After values

right Immediate value

-1 -2 -3 0 -1 -2

1 I nc 193 24 48 3 0 r nc 230 12

2 I nc 163 25

r nc 1 17 21 21 1 0

3 I nc 129 19

r nc 143 22 37 3 0

4 I nc 175 21 56 4 0 r nc 146 17

5 I nc 155 18 83 7 0 r nc 138 21

6 I nc 109 18

r nc 122 20 102 8 0

7 I nc 193 26 28 1 0 r nc 171 18

8 I nc 146 13

r nc 162 25 49 3 0 9 I nc nc 48 173 19 1 nc nc 83

10 I nc 156 13 98 7 0 nc 139 12

1 1 I nc 104 13 174 16 0 nc 177 18

12 I nc nc 67 nc 81 7 nc nc 54

13 I nc 188 34 59 3 0 nc 197 25

14 I nc 123 14 104 9 0 nc 149 18

15 I nc 223 31

nc 237 28 94 7 0

16 I nc nc 11

nc nc 83 155 13 0

17 I nc 142 1Z

nc 1 14 13 1 17 9 0

18 I nc 147 19 29 3 0 nc 210 23

19 I nc 158 18

nc 216 24 127 13 0

20 I nc 226 23 44 3 0 r nc 187 20 underlined number used for further calculations

use weighted average

nc not countable

Table 2d. List of calculated log values and log reduction factors 30.11.2000

Preparation: formulation I

Application: rubbing in for 3 min. Subject Immediate effect

No. log x logy log z

1 4.29 1.68 2.61

2 4.10 1.32 2.78

3 4.18 1.57 2.61

4 4.25 1.75 2.50

5 4.20 1.92 2.28

6 4.11 2.01 2.10

7 4.29 1.45 2.84

8 4.23 1.69 2.54

9 4.68 2.24 2.44

10 4.19 1.99 2.20

11 4.02 2.24 1.78

12 4.83 2.91 1.92

13 4.30 1.77 2.53

14 4.09 2.02 2.07

15 4.38 1.97 2.41

16 4.92 2.19 2.73

17 4.06 2.07 1.99

18 4.18 1.46 2.72

19 4.34 2.10 2.24

20 4.35 1.64 2.71

X 4.30 1.90 2.40

s 0.32 0.36 0.31

N 20 20 20

log y = log after value

s = standard deviation

log z = log-reduction factor

N = number of values (subjects) in each column

Method 2: Testing the suitability of formulation III for "Surgical hand disinfection" (DIN prEN12791 :2013)

The results are shown in Tables 3-A1 to 3-E (in Tables 3-A1 and 3-A2, germ count denotes used for the further calculation; ^* dilutions, used for the calculation). Table 3-A1

Reference product (RP): 60% (v/v) propan-1-ol)

(reference)

Application: nx3ml/3min

Date: 14.12.2015-11.1.2016

Before values Immediate after values

Subject l/r -1 -2 -3 0 -1 -2

1 ¹ >330 175 ^* 14 ^* 5 0 0 r >330 125 9

2 ¹ >330 >330 95

r >330 >330 140 >330 107 6

3 ¹ >330 124 7

r >330 151 12 22 3 0

4 ¹ 42 5 1 3 0 0 r 3 0 0

5 ¹ >330 34 3

r >330 54 8 4 1 0

6 ¹ >330 275 ^* 32 ^* 127 ^* 14 ^* 2 r 235 ^* 26 ^* 0

7 ¹ >330 >330 68

r >330 >330 138 0 0 0

8 ¹ >330 142 ^* 16 ^* >330 61 4 r >330 121 13

9 ¹ >330 >330 40 67 4 0 r >330 110 11

10 ¹ >330 >330 31

r >330 >330 35 13 1 0

11 ¹ >330 >330 81 116 13 1 r >330 >330 58

12 ¹ >330 >330 64

r >330 >330 11 2 0 0

13 ¹ >330 68 5

r >330 115 ^* 14 ^* 0 0 0

14 ¹ 10 1 0 0 0 0 r 236 ^* 18 ^* 4 15 ¹ >330 >330 54 >330 11 4 r >330 >330 138

16 ¹ >330 253 ^* 19 ^*

r >330 237 ^* 25 ^* 0 0 0

17 ¹ >330 283 ^* 27 ^* 3 0 0 r >330 221 ^* 19 ^*

18 ¹ >330 181 ^* 25 ^* 43 4 0 r >330 87 12

19 ¹ >330 157 8

r >330 37 3 0 0 0

20 ¹ >330 149 ^* 17 ^*

r >330 185 ^* 16 ^* 5 3 1

21 ¹ >330 >330 49

r >330 >330 53 >330 56 5

22 ¹ >330 >330 40 0 0 0 r >330 >330 37

23 ¹ 218 ^* 22 ^* 3

r 27 4 0 5 1 0

24 ¹ >330 >330 376 >330 158 1 1 r >330 >330 101

Table 3-A2

Test product: formulation III

Application: 2 x 5 ml / 60 s

Date: 14.12.2015-1 1 .1.2016

Before values Immediate after values

Subject l/r -1 -2 -3 0 -1 -2

1 I >330 69 7 2 0 0 r >330 47 5

2 I >330 >330 104

r >330 >330 145 >330 122 1 1

3 I >330 134 ^* 17 ^*

r 208 ^* 19 ^* 2 0 0 0

4 I >330 66 7 >330 166 ^* 15 ^* r 162 ^* 22 ^* 0 ¹ >330 >330 110

r >330 >330 232 5 0 0

¹ >330 >330 280 >330 >330 Z9 ^r >330 >330 273

¹ >330 >330 82

r >330 >330 157 41 3 0

¹ >330 252* 29 ^* 1 16 ^* 41 * 5 ^r >330 201 ^* 18 ^*

¹ >330 >330 56 58 3 0 ^r >330 330 ^* 33 ^*

¹ >330 >330 41

r >330 230 ^* 25 ^* 1 12* 15 ^* 1

¹ >330 >330 91 >330 272* 23 ^{* r} >330 >330 75

¹ >330 >330 Zl

r >330 >330 59 >330 11 8

¹ >330 185 ^* 17*

r >330 238 13 0 0 0

¹ 8 0 0 19 5 0 ^r 83 1 1 0

¹ >330 >330 135 >330 113 1 1 ^r >330 >330 99

¹ >330 >330 88

r >330 >330 60 0 0 0

¹ >330 330 ^* 31 ^* 215 ^* 24* 5 ^r >330 >330 62

¹ >330 >330 43 6 0 0 ^r >330 133 ^* 17*

¹ >330 69 4

r >330 58 1 1 1 0 0

¹ >330 >330 34

r >330 42 9 1 0 0

¹ >330 155 8

r >330 320 ^* 25 ^* 1 0 0

¹ >330 >330 40 0 0 0 ^r >330 226 ^* 29 ^* 23 I >330 84 5

r >330 11 4 0 0 0

24 I >330 >330 92 205 ^* 32 ^* 6 r >330 >330 77

Table 3-B1

List of calculated logm values and logm reductions

(Surgical hand disinfection according to prEN 12791:2013)

Reference product (RP): 60% (v/v) propan-1-ol

(reference)

Application: nx3ml/3min

Date: 14.12.2015-11.1.2016

Immediate effect

Subject n Sequence Ig before Ig after ig F values values

1 3 RP->TP 4.24 0.70 3.54

2 3 RP->TP 5.15 3.03 2.12

3 2 RP->TP 4.18 1.34 2.84

4 3 RP->TP 2.62 0.48 2.15

5 4 RP->TP 3.73 0.60 3.13

6 3 RP->TP 4.45 2.11 2.34

7 3 RP->TP 5.14 0.00 5.14

8 3 RP->TP 4.16 2.79 1.37

9 3 RP->TP 4.60 1.83 2.78

10 3 RP->TP 4.54 1.11 3.43

11 3 RP->TP 4.91 2.06 2.84

12 3 RP->TP 4.86 0.30 4.56

13 3 TP->RP 4.07 0.00 4.07

14 3 TP->RP 2.00 0.00 2.00

15 2 TP->RP 4.73 2.86 1.88

16 3 TP->RP 4.38 0.00 4.38

17 4 TP->RP 4.45 0.48 3.97

18 4 TP->RP 4.27 1.63 2.64

19 3 TP->RP 3.57 0.00 3.57

20 4 TP->RP 4.26 0.70 3.56 21 3 TP->RP 4.72 2.75 1.98

22 3 TP->RP 4.60 0.00 4.60

23 2 TP->RP 2.43 0.70 1.73

24 3 TP->RP 5.58 3.20 2.38

X 4.23 1.19 3.04 s 0.86 1.12 1.04

N 24 24 24 average

standard deviation

number of values (subjects)

number of portions of disinfectant

RF reduction factor

Table 3-B2

List of calculated logm values and logm reductions

(Surgical hand disinfection according to prEN 12791:2013)

Test product (TP): formulation III

Application: 2x5 ml/ 60s

Date: 14.12.2015-11.1.2016

Immediate effect

Subject n Sequence Ig before Ig after ig RF values values

1 2 RP->TP 3.84 0.30 3.54

2 2 RP->TP 5.16 3.09 2.08

3 2 RP->TP 3.31 0.00 3.31

4 2 RP->TP 3.82 3.22 0.60

5 2 RP->TP 5.37 0.70 4.67

6 2 RP->TP 5.45 3.90 1.55

7 2 RP->TP 5.20 1.61 3.58

8 2 RP->TP 4.41 2.15 2.25

9 2 RP->TP 4.75 1.76 2.98

10 2 RP->TP 4.37 2.06 2.30

11 2 RP->TP 4.96 3.43 1.53

12 2 RP->TP 4.77 2.86 1.91

13 2 TP->RP 4.38 0.00 4.38

14 2 TP->RP 1.90 1.28 0.62 15 2 TP->RP 5.13 3.05 2.08

16 2 TP->RP 4.78 0.00 4.78

17 2 TP->RP 4.52 2.34 2.18

18 2 TP->RP 4.63 0.78 3.86

19 2 TP->RP 3.76 0.00 3.76

20 2 TP->RP 3.62 0.00 3.62

21 2 TP->RP 4.50 0.00 4.50

22 2 TP->RP 4.60 0.00 4.60

23 2 TP->RP 3.86 0.00 3.86

24 2 TP->RP 4.96 2.33 2.63

X 4.42 1 .35 2.97

s 0.79 1 .35 1 .25

N 24 24 24

average

standard deviation

number of values (subjects)

number of portions of disinfectant

RF reduction factor

Table 3-C

Testing the sequence effects

(Surgical hand disinfection according to pREN 12791 :2013

Ig RF immediate effect

Absolute difference should be < 2

Checking the acceptance criteria according to 5.7.1. a) to e)

a) A complete set of results from 24 subjects is available (accordingly equivalent or more than the minimum number of 23); b) average of the Ig before values for RP = 4.23/4.15 and for TP = 4.42/4.43 (accordingly all greater than 3.5);

c) number of individual Ig reductions less than 2.00 in the case of RP (immediate effect = 4 (accordingly less than 1 1 );

d) immediate effect: in the case of the group with the sequence RP->TP the difference of Ig RF is: 3.02 - 2.53 = 0.49, in the case of the group with the sequence TP->RP the difference of Ig RF is: 3.06 - 3.41 = 0.34, the absolute difference in the average differences is: abs[(0.49)-(0.34)] = 0.84 (accordingly less than 2.00)

e) all quotients of weighted average germ counts are between 5 and 15.

All acceptance criteria are satisfied.

Table 3-D

Calculation of the individual differences of Ig RF of RP - TP

(Surgical hand disinfection according to prEN 12791:2013)

Ig RF immediate effect

Subject RP Difference Difference

RP-TP RP-TP

1 3.54 3.54 0.00

2 2.12 2.08 0.04

3 2.84 3.31 -0.48

4 2.15 0.60 1.54

5 3.13 4.67 -1.54

6 2.34 1.55 0.79

7 5.14 3.58 1.56

8 1.37 2.25 -0.88

9 2.78 2.98 -0.21

10 3.43 2.30 1.13

11 2.84 1.53 1.31

12 4.56 1.91 2.64

13 4.07 4.38 -0.31

14 2.00 0.62 1.38

15 1.88 2.08 -0.20

16 4.38 4.78 -0.40

17 3.97 2.18 1.79

18 2.64 3.86 -1.22

19 3.57 3.76 -0.20

20 3.56 3.62 -0.06

21 1.98 4.50 -2.52

22 4.60 4.60 0.00

23 1.73 3.86 -2.12

24 2.38 2.63 -0.25 Table 3-E

Sorting the individual differences and calculation of the upper 97.5% confidence limits according to Hodges- Lehmann

(Surgical hand disinfection according to prEN 12791 :2013)

Immediate effect

16 -0.25 1.19 ³ ' 0.77 ^ei 0.65 ⁸² 0.64 ⁸³ 0.56 0.53 0.44 0.27 -0.1 1 -0.13 -0.13

17 -0.31 1.17 ^3a 0.74 ^e4 0.62 0.62 0.53 0.50 0.41 0.24

18 -0.40 1.12 ⁴ 0.70 ^DB 0.58 0.57 0.49 0.46 0.36 0.19

19 -0.48 1.08 ⁴⁴ 0.66 ^BU 0.54 0.53 0.45 0.42 0.32 0.16

20 -0.88 0.88 ^au 0.46 0.34 0.33 0.25 0.22 0.12 -0.05

21 -1.22 0.71 ^DD 0.29 0.17 0.16 0.08 0.05 -0.04

22 -1.54 0.55 0.13 0.01 0.00 -0.08 -0.1 1

23 -2.12 0.26

24 -2.52 0.06

Tab. 3-F. Sign ranking test for paired samples

(Surgical hand disinfection according to prEN 12791 :2013)

The median is between the 12 ^th and 13 ^th value [-0.06 + (-0.20)]/2 = -0.13. The small exponents indicate the rankings. The average pairwise differences, which do not exceed the median value (here: -0.13), are calculated. Table 3F of the critical values for sign ranking tests for paired samples according to Wilcoxon reveals, in the case of an input value of N=24 and a one-tailed significance level of 0.025, a critical value of 81 . Consequently, c = 81 +1 = 82. The pairwise differences are sorted in descending order. The 82 ^nd value is 0.65. Consequently, the one-tailed upper 97.5% confidence limit for the difference of the Ig RF between RP and TP is 0.65, which is lower than the agreed limit for inferiority of 0.75. Consequently, the hypothesis of the inferiority of the immediate effect of TP is discarded and the conclusion can be drawn that the preparation to be tested (TP) is not inferior to the reference product (RP).

SUMMARY OF THE RESULTS AND DISCUSSION a) EN 13727

Surprisingly, it has been possible to show that the use according to the invention of fatty alcohols in combination with the alcohol constituents (isopropanol and ethanol) leads to a marked increase in efficacy (see Tab. 4, quantitative suspension experiment according to EN 13727, test organism Escherichia coii under clean conditions). Tab. 4. Quantitative suspension experiment according to EN 13727, E. coli, clean conditions: comparison of the influence of different refatting agent systems on the microbiological efficacy

Comparison

It was surprising that the formulation III according to the invention, which does not comprise 1 -(2-ethylhexyl) glycerol ether, has a similarly good microbiological efficacy to a comparable standard commercial formulation II with a content of 1 -(2- ethylhexyl) glycerol ether. b) EN12791

The formulation III led in the experiments under conditions simulating those in practice of a surgical hand disinfection with an application time of 60 seconds to the following results (cf. Tables 3A1 -3B2):

Tab. 5. Summary of the results of Tables 3A1 -3B2

The difference between the average log-io reduction factors (RF) of the two methods was tested by means of the Hodges & Lehmann test (Tables 3D-3F). The one-tailed upper 97.5% confidence limit for the difference in log-io RF between the reference and the test product is 0.65 for the immediate effect, which is lower than the agreed limit for inferiority of 0.75. Formulation III therefore proved itself to be not inferior in its immediate effect compared to the reference. Consequently, the efficacy of formulation III during subsequent use for surgical hand disinfection confirms: 2 x 5 ml / 60 seconds.

The microbiological efficacy is thus considerably superior in the case of the use according to the invention of fatty alcohols, compared to formulation I that is commercially customary for surgical hand disinfection (EN 12791 , methodology according to the particular valid version, see Tab. 6).

Tab. 6: Comparison of the efficacy in surgical hand disinfection (efficacy according to EN 12791 , respective valid version)

^* comparison

Thus, in the case of the use according to the invention of fatty alcohols (in formulation III), an efficacy of 60 seconds in surgical hand disinfection was demonstrated, whereas formulation I, a standard commercial preparation, was effective only in 180 seconds.