EP4291626A1

EP4291626A1 - Anti-microbial composition

Info

Publication number: EP4291626A1
Application number: EP21712412.2A
Authority: EP
Inventors: Liangjing FANG; Yuxiang Liu; Xiujun Xu
Original assignee: Procter and Gamble Co
Current assignee: Procter and Gamble Co
Priority date: 2021-02-09
Filing date: 2021-02-09
Publication date: 2023-12-20
Also published as: CN114903039A; WO2022170454A1; JP2023550186A

Abstract

A composition comprising anti-microbial composition comprising: a) 4-chloro-3, 5-dimethyl phenol; and b) a hydroxyl diphenyl ether.

Description

ANTI-MICROBIAL COMPOSITION

FIELD OF THE INVENTION

The present invention relates to an anti-microbial composition comprising 4-chloro-3, 5-dimethyl phenol and a hydroxyl diphenyl ether.

BACKGROUND OF THE INVENTION

Consumer products have evolved to address user needs for an anti-microbial benefit. Particularly, in the field of laundry products, consumer needs in the term of cleaning have been well met, but there are still some other unmet consumer needs including anti-microbial benefit. For example, an anti-microbial laundry product is desired by users as it can deliver an anti-microbial benefit on fabrics while washing. Currently, various anti-microbial agents, e.g., bleaching agents, 4-chloro-3, 5-dimethyl phenol (also called Chloroxylenol or PCMX) , Benzalkonium Chloride (BKC) , diphenyl ethers, are known for use in consumer product formulations to deliver an anti-microbial effect. Particularly, PCMX and BKC can deliver a benefit of microbial removal during wash, and diphenyl ethers can deliver a benefit of microbial prevention during storage or use of fabrics.
However, in the context of liquid products, it is still challenging to achieve a desired anti-microbial efficacy. Particularly, it is desirable for consumers that laundry products can achieve both microbial removal (which means removing microorganisms on fabrics during wash) and microbial prevention (which means preventing microorganisms during storage or use) . However, it is very challenging to deliver both microbial removal and microbial prevention at the same time because of incompatibility between different actives. Particularly, an active for microbial removal might significantly compromise the efficacy of microbial prevention by an active for microbial prevention through interaction therebetween.
Further, another challenge for formulating a liquid laundry product containing an active for microbial prevention during storage or use is how to ensure a sufficient amount of such active deposited onto washed fabrics. This is very challenging because various components commonly used in laundry detergent products may significantly compromise the deposition of such anti-microbial agent on fabrics.
Thus, there is a continuing need for laundry compositions that enable improved anti-microbial efficacy, preferably both in the aspects of microbial removal and microbial prevention.
SUMMARY OF THE INVENTION
It is a surprising and unexpected discovery of the present invention that the anti-microbial composition according to the present disclosure can meet the need as above, i.e., the anti-microbial composition according to the present disclosure can deliver both an improved efficacy of microbial removal and an improved efficacy of microbial prevention.
In one aspect, the present invention provides an anti-microbial composition comprising: a) 4-chloro-3, 5-dimethyl phenol; and b) hydroxyl diphenyl ether of formula (I) :
wherein:
each Y is independently selected from chlorine, bromine, or fluorine,
each Z is independently selected from SO2H, NO2, or C1-C4 alkyl,
r is 0, 1, 2, or 3,
o is 0, 1, 2, or 3,
p is 0, 1, or 2,
m is 1 or 2, and
n is 0 or 1;
wherein 4-chloro-3, 5-dimethyl phenol is present in an amount sufficient for delivering a Through-The-Wash (TTW) dosage ranging from 20 ppm to 960 ppm, preferably from 40 ppm to 800 ppm, more preferably from 80 ppm to 400 ppm, yet more preferably from 120 ppm to 300 ppm, most preferably from 160 ppm to 240 ppm.
Surprisingly, when the TTW dosage of 4-chloro-3, 5-dimethyl phenol in the formulation containing 4-chloro-3, 5-dimethyl phenol and a hydroxyl diphenyl ether is within a certain range, e.g. from 20 ppm to 960 ppm, such formulation can deliver both the efficacies of microbial removal and microbial prevention. In the contrast, if the TTW dosage of 4-chloro-3, 5-dimethyl phenol is more than 960 ppm, the efficacy provided by the hydroxyl diphenyl ether would be significantly compromised. Without being bound by any theory, it is believed that an overdose of 4-chloro-3, 5-dimethyl phenol might interfere with the deposition of the hydroxyl diphenyl ether onto fabrics, resulting in the compromised microbial prevention.
Preferably, the composition according to the present invention is a liquid composition, such as a liquid sanitizer.
Further, it is even more surprising that when the TTW dosage of 4-chloro-3, 5-dimethyl phenol in the formulation containing 4-chloro-3, 5-dimethyl phenol and a hydroxyl diphenyl ether is within the preferred range, e.g. from 20 ppm to 960 ppm, the deposition of the hydroxyl diphenyl ether is significantly improved, which is completely unexpected.
Preferably, the hydroxyl diphenyl ether is present in an amount sufficient for delivering a Through-The-Wash (TTW) dosage ranging from 0.1 ppm to 400 ppm, preferably from 0.5 ppm to 100 ppm, more preferably from 1 ppm to 20 ppm, yet more preferably from 1.5 ppm to 12 ppm, most preferably from 2 ppm to 8 ppm.
In some preferred embodiments, the weight ratio of 4-chloro-3, 5-dimethyl phenol to the hydroxyl diphenyl ether in the composition according to the present disclosure may be between 0.1 and 2000, preferably between 1 and 1000, more preferably between 5 and 200, yet more preferably between 30 and 70, yet more preferably between 35 and 65, yet more preferably between 40 and 60, and most preferably between 45 and 55.
In some preferred embodiments, the hydroxyl diphenyl ether may be selected from the group consisting of 4-4’-dichloro-2-hydroxy diphenyl ether, 2, 4, 4’-trichloro-2’-hydroxy diphenyl ether, and a combination thereof, preferably 4-4’-dichloro-2-hydroxy diphenyl ether.
Preferably, 4-chloro-3, 5-dimethyl phenol is present in an amount ranging from 0.1%to 4.8%, preferably from 0.2%to 4%, more preferably from 0.4%to 2%, yet more preferably from 0.6%to 1.5%, most preferably from 0.8%to 1.2%, by weight of the composition; and/or the hydroxyl diphenyl ether is present in an amount ranging from 0.0005%to 2%, preferably from 0.0025%to 0.5%, more preferably from 0.005%to 0.1%, yet more preferably from 0.0075%to 0.06%, most preferably from 0.01%to 0.04%, by weight of the composition.
In some particular embodiments, the composition according to the present disclosure may further comprise from 1%to 50%, more preferably from 3%to 40%, most preferably from 5%to 30%, by total weight of the composition, of a solvent. Preferably, the solvent may be an alcohol selected from the group consisting of ethanol, ethylene glycol, propylene glycol, glycerol, butanediol, butanetriol, poly (propylene glycol) , terpineol, and a combination thereof. More preferably, the solvent may be propylene glycol and/or terpineol.
In some particular embodiments, the composition according to the present disclosure may further comprise from 0.1%to 30%, more preferably from 0.5%to 20%, most preferably from 1%to 10%, by total weight of the composition, of a crystalline, hydroxyl-containing stabilizing agent. Preferably, the stabilizing agent may be selected from the group consisting of microcrystalline cellulose or derivatives thereof, castor oil or derivatives thereof, hydrogenated castor oil or derivatives thereof, and any combinations thereof. More preferably, the stabilizing agent may be castor oil.
In some particular embodiments, the composition according to the present disclosure may further comprise from 0.01%to 5%, preferably from 0.02%to 2%, more preferably from 0.05%to 1%, of a perfume by total weight of the composition. Preferably, the perfume may be selected from the group consisting of free perfumes, pro-perfumes, encapsulated perfumes, perfume microcapsules, and combinations thereof. More preferably, the perfume may comprise a combination of free perfumes and perfume microcapsules. Most preferably, the weight ratio of free perfumes to perfume microcapsules in the composition may be from 1: 5 to 20: 1, preferably from 1: 2 to 10: 1, more preferably from 1: 1 to 5: 1, most preferably from 1.5: 1 to 3: 1.
In some particular embodiments, the composition is a sanitizer.
In another aspect, the present invention relates to a process for treating an article of clothing comprising the steps of:
a) providing an article of clothing and an anti-microbial composition according to the present disclosure, and
b) contacting the article of clothing with the composition, preferably for at least 5 min, such as 5 min, 10 min, 15 min, 20 min, 30 min, 50 min, 100 min, 200 min, 500 min, 1000 min or any ranges therebetween.
In another aspect, the present invention relates to a process for treating an article of clothing comprising the steps of:
a) providing an article of clothing and an anti-microbial composition according to the present disclosure in a container, e.g. a laundry basin;
b) contacting the article of clothing with the composition in the container for a certain period, e.g. at least 5 min, at least 10 min, at least 15 min or at least 20 min; and
c) washing the article of clothing by hand or in a washing machine.
It is an advantage of the composition according to the present disclosure that it may deliver efficacies in the both aspects of microbial removal and microbial prevention. In the context of the present disclosure, removing microorganism includes but not limited to kill, deactivate, eliminate and/or wash away microorganism.
It is another advantage of the composition according to the present disclosure that it may improve the microbial prevention without compromising the microbial removal.
It is another advantage of the composition according to the present disclosure that it may improve the deposition of the hydroxyl diphenyl ether on fabrics.
These and other aspects of the present invention will become more apparent upon reading the following detailed description of the invention.

DETAILED DESCRIPTION OF THE INVENTION

Features and benefits of the various embodiments of the present invention will become apparent from the following description, which includes examples of specific embodiments intended to give a broad representation of the invention. Various modifications will be apparent to those skilled in the art from this description and from practice of the invention. The scope of the present invention is not intended to be limited to the particular forms disclosed and the invention covers all modifications, equivalents, and alternatives falling within the spirit and scope of the invention as defined by the claims.
The dimensions and values disclosed herein are not to be understood as being strictly limited to the exact numerical values recited. Instead, unless otherwise specified, each such dimension is intended to mean both the recited value and a functionally equivalent range surrounding that value. For example, a dimension disclosed as “40 mm” is intended to mean “about 40 mm. ”
As used herein, terms such as "a" and "an" when used in a claim, are understood to mean one or more of what is claimed or described. The terms “comprise, ” “comprises, ” “comprising, ” “contain, ” “contains, ” “containing, ” “include, ” “includes” and “including” are all meant to be non-limiting.
As used herein, the term “liquid composition” herein refers to compositions that are in a form selected from the group consisting of pourable liquid, gel, cream, and combinations thereof. The liquid composition may be either aqueous or non-aqueous, and may be anisotropic, isotropic, or combinations thereof.
The term “consisting essentially of” means that the composition contains less than about 10%, preferably less than about 5%, of ingredients other than those listed.
Further, the term “substantially free of” or “substantially free from” means that the indicated material is present in the amount of from 0 wt%to about 1 wt%, preferably from 0 wt% to about 0.5 wt%, more preferably from 0 wt%to about 0.2 wt%. The term “essentially free of” means that the indicated material is present in the amount of from 0 wt%to about 0.1 wt%, preferably from 0 wt%to about 0.01 wt%, more preferably it is not present at analytically detectable levels.
As used herein, the term “washing solution” refers to the typical amount of aqueous solution used for one cycle of laundry washing, preferably from 1 L to 50 L, alternatively from 1 L to 20 L for hand washing and from 20 L to 50 L for machine washing.
As used herein, the term “soiled fabric” is used non-specifically and may refer to any type of natural or artificial fibers, including natural, artificial, and synthetic fibers, such as, but not limited to, cotton, linen, wool, polyester, nylon, silk, acrylic, and the like, as well as various blends and combinations.
As used herein, all concentrations and ratios are on a weight basis unless otherwise specified. All temperatures herein are in degrees Celsius (℃) unless otherwise indicated. All conditions herein are at 20℃ and under the atmospheric pressure, unless otherwise specifically stated. All polymer molecular weights are determined by weight average number molecular weight unless otherwise specifically noted.
Composition
The composition of the present invention comprises 4-chloro-3, 5-dimethyl phenol and a hydroxyl diphenyl ether. It has been surprisingly found that, by combining 4-chloro-3, 5-dimethyl phenol and a hydroxyl diphenyl ether at a certain range of weight ratio therebetween, an excellent efficacy in the aspects of microbial removal and microbial prevention can be achieved.
Preferably, 4-chloro-3, 5-dimethyl phenol may be present in an amount sufficient for delivering a Through-The-Wash (TTW) dosage ranging from 20 ppm to 960 ppm, preferably from 40 ppm to 800 ppm, more preferably from 80 ppm to 400 ppm, yet more preferably from 120 ppm to 300 ppm, most preferably from 160 ppm to 240 ppm, for example 30 ppm, 40 ppm, 50 ppm, 60 ppm, 70 ppm, 80 ppm, 90 ppm, 100 ppm, 110 ppm, 120 ppm, 130 ppm, 140 ppm, 150 ppm, 160 ppm, 170 ppm, 180 ppm, 190 ppm, 200 ppm, 210 ppm, 220 ppm, 230 ppm, 240 ppm, 250 ppm, 260 ppm, 270 ppm, 280 ppm, 290 ppm, 300 ppm or any ranges therebetween.
Preferably, the hydroxyl diphenyl ether may be present in an amount sufficient for delivering a Through-The-Wash (TTW) dosage ranging from 0.1 ppm to 400 ppm, preferably from 0.5 ppm to 100 ppm, more preferably from 1 ppm to 20 ppm, yet more preferably from 1.5 ppm to 12 ppm, most preferably from 2 ppm to 8 ppm, for example 0.1 ppm, 0.2 ppm, 0.3 ppm, 0.4 ppm, 0.5 ppm, 0.6 ppm, 0.7 ppm, 0.8 ppm, 0.9 ppm, 1 ppm, 1.1 ppm, 1.2 ppm, 1.3 ppm, 1.4 ppm, 1.5 ppm, 1.6 ppm, 1.7 ppm, 1.8 ppm, 1.9 ppm, 2 ppm, 3 ppm, 4 ppm, 5 ppm, 6 ppm, 7 ppm, 8 ppm, 9 ppm, 10 ppm, 11 ppm, 12 ppm, 15 ppm, 20 ppm, 30 ppm, 35 ppm, 40 ppm, 45 ppm, 50 ppm, 60 ppm, 70 ppm, 80 ppm, 90 ppm, 100 ppm or any ranges therebetween.
In some preferred embodiments, the weight ratio of 4-chloro-3, 5-dimethyl phenol to the hydroxyl diphenyl ether may be between 0.1 and 2000, preferably between 1 and 1000, more preferably between 5 and 200, yet more preferably between 30 and 70, yet more preferably between 35 and 65, yet more preferably between 40 and 60, and most preferably between 45 and 55, for example 30, 35, 40, 45, 50, 55, 60, 65, 70 or any ranges therebetween.
Preferably, 4-chloro-3, 5-dimethyl phenol may be present in an amount ranging from 0.2%to 4.8%, preferably from 0.4%to 2%, more preferably from 0.6%to 1.5%, most preferably from 0.8%to 1.2%, for example 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%or any ranges therebetween, by weight of the composition; and/or the hydroxyl diphenyl ether may be present in an amount ranging from 0.0025%to 0.5%, preferably from 0.005%to 0.1%, more preferably from 0.0075%to 0.06%, most preferably from 0.01%to 0.04%, for example 0.0025%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.012%, 0.015%, 0.017%, 0.02%, 0.025%, 0.03%, 0.035%, 0.04%, 0.045%, 0.05%, 0.055%, 0.06%, 0.065%, 0.07%, 0.075%, 0.08%, 0.085%, 0.09%, 0.095%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%or any ranges therebetween, by weight of the composition.
The laundry composition herein provides efficacy for removing Gram positive bacteria (e.g., Staphylococcus aureus) and/or Gram negative bacteria (e.g., E. coli) . In one embodiment, the laundry composition provides a Microbial Removal Value of at least a log 1.0 reduction, preferably at least a log 1.5 reduction, more preferably at least a log 2.0 reduction, yet more preferably a log 2.5 reduction, yet more preferably a log 3.0 reduction, most preferably a log 3.5 reduction, against Gram positive bacteria and/or Gram negative bacteria, to treated fabrics versus non-treated fabrics.
Further, the laundry composition herein provides efficacy for microbial prevention to the fabrics treated by the composition. In one embodiment, the laundry composition provides a Bacteriostatic Activity Value of at least a log 1.0 reduction, preferably at least a log 1.5 reduction, more preferably at least a log 2.0 reduction, yet more preferably a log 2.5 reduction, against Gram positive bacteria and/or Gram negative bacteria, to treated fabrics versus non-treated fabrics. Preferably, the composition provides at least a log 1.0 reduction, preferably at least a log 1.5 reduction, more preferably at least a log 2.0 reduction, yet more preferably a log 2.5 reduction, against E. coli, Staphylococcus aureus and/or Klebsiella pneumoniae after a 10 minutes contact time in a 1055 ppm aqueous solution as determined by the QB/T 2738 7.2 suspension method (that is described below) .
Hydroxyl diphenyl ether
The hydroxyl diphenyl ether useful in the composition according to the present invention is a nonionic compound.
Preferably, the hydroxyl diphenyl ether can be either halogenated or non-halogenated, but preferably is halogenated. In one embodiment, the hydroxyl diphenyl ether is a hydroxyl diphenyl ether of formula (I) :
wherein:
each Y is independently selected from chlorine, bromine, or fluorine, preferably is chlorine or bromine, more preferably is chlorine,
each Z is independently selected from SO ₂H, NO ₂, or C ₁-C ₄ alkyl,
r is 0, 1, 2, or 3, preferably is 1 or 2,
o is 0, 1, 2, or 3, preferably is 0, 1 or 2,
p is 0, 1, or 2, preferably is 0,
m is 1 or 2, preferably is 1, and
n is 0 or 1, preferably is 0.
In the above definition for formula (I) , 0 means nil. For example, when p is 0, then there is no Z in formula (I) . Each Y and each Z could be the same or different. In one embodiment, o is 1, r is 2, and Y is chlorine or bromine. This embodiment could be: one chlorine atom bonds to a benzene ring while the bromine atom and the other chlorine atom bond to the other benzene ring; or the bromine atom bonds to a benzene ring while the two chlorine atoms bond to the other benzene ring.
More Preferably, the hydroxyl diphenyl ether is selected from the group consisting of 4-4’-dichloro-2-hydroxy diphenyl ether ( “Diclosan” ) , 2, 4, 4’-trichloro-2’-hydroxy diphenyl ether ( “Triclosan” ) , and a combination thereof. Most preferably, the hydroxyl diphenyl ether is 4-4’- dichloro-2-hydroxy diphenyl ether, commercially available from BASF, under the trademark name HP 100.
4-chloro-3, 5-dimethyl phenol (PCMX)
4-chloro-3, 5-dimethyl phenol, also known as chloroxylenol or para-chloro-meta-xylenol (PCMX) , is an antiseptic and disinfectant which is used for skin disinfection and cleaning surgical instruments. It has been also used in laundry applications. It has the following formula (II) :
Solvent
The laundry composition herein may further comprise from 1%to 50%, more preferably from 3%to 40%, most preferably from 5%to 30%, by total weight of the composition, of a solvent. Preferably, the solvent may be an alcohol selected from the group consisting of ethanol, ethylene glycol, propylene glycol, glycerol, butanediol, butanetriol, poly (propylene glycol) , terpineol, and a combination thereof. More preferably, the solvent may be propylene glycol and/or terpineol.
In some particular embodiments, the presence of the solvent may provide additional benefit (s) , for example improved stability.
Stabilizing agent
The laundry composition herein may further comprise from 0.1%to 30%, more preferably from 0.5%to 20%, most preferably from 1%to 10%, by total weight of said composition, of a crystalline, hydroxyl-containing stabilizing agent. Ppreferably, the stabilizing agent may be selected from the group consisting of microcrystalline cellulose or derivatives thereof, castor oil or derivatives thereof, hydrogenated castor oil or derivatives thereof, and any combinations thereof. More preferably, the stabilizing agent may be castor oil.
In some particular embodiments, the presence of the solvent may provide additional benefit (s) , for example improved stability.
Adjunct Ingredients
The laundry composition herein may further comprise adjunct ingredients. Suitable adjunct materials include but are not limited to: builders, chelating agents, rheology modifiers, dye transfer inhibiting agents, dispersants, enzymes, and enzyme stabilizers, catalytic materials, bleach activators, hydrogen peroxide, sources of hydrogen peroxide, preformed peracids, polymeric dispersing agents, clay soil removal/anti-redeposition agents, brighteners, suds suppressors, dyes, photobleaches, perfumes, structure elasticizing agents, fabric softeners, carriers, hydrotropes, processing aids, solvents, hueing agents, structurants and/or pigments. The precise nature of these adjunct ingredients and the levels thereof in the laundry composition will depend on the physical form and/or the use of the composition.
Preferably, the composition according to the present disclosure may further comprise from 0.01%to 5%, preferably from 0.02%to 2%, more preferably from 0.05%to 1%, of a perfume by total weight of said composition; wherein said perfume is preferably selected from the group consisting of free perfumes, pro-perfumes, encapsulated perfumes, perfume microcapsules, and combinations thereof; wherein said perfume more preferably comprises a combination of free perfumes and perfume microcapsules; wherein most preferably the weight ratio of free perfumes to perfume microcapsules in said composition is from 1: 5 to 20: 1, preferalby from 1: 2 to 10: 1, more preferably from 1: 1 to 5: 1, most preferably from 1.5: 1 to 3: 1.
Composition Preparation
The laundry composition of the present invention is generally prepared by conventional methods such as those known in the art of making laundry compositions. Such methods typically involve mixing the essential and optional ingredients in any desired order to a relatively uniform state, with or without heating, cooling, application of vacuum, and the like, thereby providing laundry compositions containing ingredients in the requisite concentrations.
Method of Use
Another aspect of the present invention is directed to a method of using the laundry composition to treat a fabric with a microbial removal benefit and a microbial preventive benefit. The method comprises the step of administering from 1 g to 2000 g of the above-mentioned laundry composition into a laundry washing basin comprising water to form a soaking solution. The soaking solution in a laundry washing basin herein preferably has a volume from 1 L to 50 L, alternatively from 1 L to 20 L for hand washing and from 20 L to 50 L for machine washing. Preferably, the microbial removal benefit herein is determined by the method as described in Test 1 (QB/T 2738 7.2 Germ kill by suspension) and the microbial preventive benefit herein is determined by the method as described in Test 2 (GB/T 20944.2 Absorption method) . The temperatures of the laundry washing solution preferably range from 5℃ to 60℃.
The dosing amount in the method herein may be different depending on the washing type. In one embodiment, the method comprises administering from about 20 g to about 100 g of the composition into a hand washing basin (e.g., about 1-5 L) . In an alternative embodiment, the method comprises administering from about 100 g to about 1000 g, preferably from about 600 g to about 900 g of the composition into a washing machine (e.g., about 30-45 L) .
Preferably, the method herein further comprises the step of contacting a soiled fabric with the soaking solution. For example, the presence of Gram-positive bacteria and/or Gram-negative bacteria is suspected on the fabric. The step of contacting the soiled fabric with the soaking solution is preferably after the step of administering the composition in a laundry washing basin.
Test Method
Test 1: Efficacy of Microbial Removal (QB/T 2738 7.2 suspension method)
The efficacy of microbial removal for laundry compositions is determined by the method as defined in QB/T 2738 7.2 suspension method and described hereinafter.
1. Microorganism Preparation:
A. Subculture microorganism (s) on Nutrient Agar through at least one daily transfer, incubating at 35 ± 2℃.
B. On the day prior to testing, transfer the cells into another Nutrient Agar. Incubate 18 to 24 h at 35 ± 2℃, agar side down.
C. Remove growth from the agar plate using three-mL dilution fluid and five sterile glass beads to suspend growth. The cultures will be standardized to yield required colony forming units (CFU) per mL of S. aureus , K. pneumoniae and E. coli.
2. Procedure:
A. Dilute the test sample based on recommended dosage. Use sterile container. Mix thoroughly. Test material can be mixed using a sterile magnetic stir bar and a magnetic stir plate.
B. Add 5 mL of the test material to a centrifuge tube.
C. At time 0, add 0.1 mL of inoculum prepared in Step 1C) to the test material prepared in 2B) , mix well. Start the timer immediately.
D. React till the defined time, pipette 0.5ml of above solution into 4.5ml nertralizer and mixe well. Leave to neutralize 10mins.
E. Make a proper dilution and take 1ml for enumeration in duplicate using standard plating techniques.
F. Incubate at the appropriate temperature, time, and environment for the test organism.
G. Count colonies using standard plate count rules and record raw data as CFU/plate. Average duplicate plates and multiply by the dilution factor to calculate CFU/mL.
H. Replace product solution by PBS and repeat above procedures as control.
I. Each sample, time point, and organism shall be run in triplicate.
3. Calculation of Microbial Removal Activity Value:
Microbial Removal Activity (%) = (Average bacteria amount of Control –Average bacteria amount of Sample) /Average bacteria amount of Control
Microbial Removal Activity (LogR) = -Log ₁₀ (1-Microbial Removal Activity (%) )
A Microbial Removal Activity Value (LogR) of no less than 1.0 represents acceptable microbial removal efficacy. A Microbial Removal Activity Value (LogR) of no less than 2.0 represents good microbial removal efficacy. And a Microbial Removal Activity Value (LogR) of lower than 1.0 indicates unacceptable poor microbial removal efficacy.
Test 2: Efficacy of Microbial Prevention (GB/T 20944.2)
The efficacy of microbial prevention for laundry compositions is determined by the method as defined in the GB/T 20944.2 Absorption method and described hereinafter.
1. Microorganism Preparation:
A. Aseptically add certain amount of nutrient broth into a lyophilized culture of Staphylococcus aureus, Escherichia coli or Klebsiella pneumoniae. Dissolve and suspend the culture in the nutrient broth to obtain a suspension. Streak a loop of the suspension onto a nutrient agar plate, and incubate at 37℃ for 24 hours to obtain a first generation subculture of bacterial suspension. Transfer a colony of the first generation subculture of bacterial suspension into 20 mL of nutrient broth with shaking, and incubate at 37℃ for 24 hours to obtain a second generation subculture of bacterial suspension. Transfer 0.4 mL of the second generation subculture of bacterial suspension into another 20 mL of nutrient broth with shaking, and incubate at 37℃ for 3±1 hours to obtain a third generation subculture of bacterial suspension.
B. Dilute the third generation subculture of bacterial suspension by 1/20 diluted nutrient broth to a concentration of 1×10 ⁵ cfu/mL to 3x10 ⁵ cfu/ml to obtain a working culture.
C. Store the working culture at 4℃ and use within 4h.
2. Fabric washing:
A. Boil two fabric strips each having a width of 1 m and length of 3 m (32 yarn/cm×32 yarn/cm, 100%plain weave cotton) in 5 L of a solution for 1 hour. The solution is prepared by 2.5 g of a nonionic soaked agent, 2.5 g of sodium carbonate, and 5000 mL of distilled water. The nonionic soaked agent is prepared by 5.0 g of alkylphenol ethoxylate, 5 g of sodium carbonate, and 1000 mL of distilled water. Rinse the fabric strips in boiled deionized water for 5 minutes. Place the fabric strips in cool deionized water for 5 minutes, and indoor dry.
B. Fix one end of the test fabric strip obtained from step 2A onto a stainless steel spindle at an outer position along the horizontal extension of the stainless steel spindle. The stainless steel spindle has 3 horizontal stands that are connected to one another. Wrap the test fabric strip around the 3 horizontal stands of the stainless steel spindle with sufficient tension to obtain a fabric wrapped spindle having 12 laps of fabric. Fix the other end of the test fabric strip onto the outer lap of the 12 laps of fabric via a pin. Sterilize the fabric wrapped spindle with pressure steam at 121℃ for 15 minutes.
C. Dissolve 3.4 g of calcium chloride dihydrate and 13.9 g of magnesium chloride hexahydrate in 100 mL of distilled water, and then sterilize the mixture with pressure steam at 121℃ for 20 minutes. Add 1 mL of the mixture into 1L of distilled water to obtain a hard water solution.
D. Add sufficient amount of sample into 1L of the hard water solution obtained from step 2C to obtain a solution having required concentration. Mix the solution by a magnetic stirrer for 4 minutes. Distribute 265 mL of the mixed solution into an exposure chamber to obtain a washing solution. Place the exposure chamber in a water bath and achieve the test temperature of (25±1) ℃.
E. Aseptically soak the fabric wrapped spindle obtained from step 2B into the washing solution in the exposure chamber, and close the exposure chamber with a lid.
F. Fix the exposure chamber onto a tumbler. Soaking 8mins and then rotate the tumbler for 16 minutes. Then remove the fabric wrapped spindle from the exposure chamber. Place the fabric wrapped spindle in Haier iwash-1p Top Load Washing Machine and spin-dry for 3 minutes.
G. Discard the washing solution from the exposure chamber, and then add 265 mL of sterilized distilled water into the exposure chamber. Soak the spin-dried fabric wrapped spindle in the newly added distilled water in the exposure chamber. Rotate the tumbler for 3 minutes and spin-dry for 3 minutes.
H. Repeat step 2G.
I. Aseptically remove the fabric wrapped spindle out of the exposure chamber and remove the test fabric strip from the spindle. Air dry the test fabric strip overnight.
3. Fabric Incubation:
A. Cut the washed test fabric strip obtained from step 2I to square pieces having a side length of 2 cm. Obtain 3 test specimens with a mass of 0.40g±0.05g for the following steps. Also obtain 6 test specimens in a same way by using the fabric from step 2B as Control.
B. Put each set of specimens into a vial, and then sterilize the specimens with pressure steam at 121℃ for 15 minutes. After the sterilization, dry the specimens for 1 hour in a clean bench without a cap.
C. Inoculate 0.2 mL of the working culture obtained from step 1B onto each dried specimen. Immediately after the inoculation, extract the bacteria on 3 Control test specimen, plate with nutrient agar and incubate at 37℃ for 24-48 hours. Count the total colony-forming units (CFU) of each set of specimens, and obtain average results of the 3 sets. Take the log10 value of CFU value as C ₀. Incubate other 6 vials containing the inoculated specimens at 37℃ for 18 to 24 hours.
D. Extract survivors on the incubated specimens, plate with nutrient agar, and incubate at 37℃ for 24-48 hours. Count the total colony-forming units (CFU) of each set of specimens, and obtain average results of the 3 sets. Take the log10 value of average CFU value as C _t (for Control) or T _t (for test sample) .
4. Calculation of Microbial Prevention Activity Value:
Microbial Prevention Activity Value (%) = (Average CFU of Control –Average CFU of Sample) /Average CFU of Control
Microbial Prevention Activity Value (logR) = C _t – T _t
A Microbial Prevention Activity Value (logR) of no less than 1.0 represents acceptable microbial preventive efficacy, and of no less than 2.0 represents excellent microbial preventive efficacy. And a Microbial Prevention Activity Value (logR) of lower than 1.0 indicates unacceptable poor microbial preventive efficacy.
Test 3: Fabric Deposition Test for Anti-Microbial Agents
Anti-microbial agents are extracted from treated fabrics by using the methanol-based Accelerated Solvent Extraction (ASE) method described hereinafter. The resulting extract is then subjected to gradient reversed-phase high performance liquid chromatographic (HPLC) separation on a C18 column and is quantified by tandem mass spectrometry (MS/MS) operating under multiple reaction monitoring (MRM) conditions at negative mode.
As a first step, about three (3) grams of the treated fabrics is accurately weighed and then filled into a steel ASE tube. The extraction protocol is run for about five (5) minutes using methanol as the extraction solvent at an elevated temperature of about 100℃ and a pressure of about 2000 pound per square inch (psi) . The resulting extract is collected and transferred into a 25-ml flask, which is then brought to volume with methanol. The resulting solution is then diluted by about twenty-five (25) fold by using a mixture of methanol and water at a 50: 50 ratio, which is used as an injection sample for the subsequent LC-MS/MS analysis.
Next, about five (5) ul of the above-mentioned injection sample is injected and separated on a Waters ACQUITY UPLC C18 column with gradient from about 70%mobile phase A (1%formic water solution) /30%mobile phase B (0.1%formic acid in methanol) to 5%mobile phase A/95%mobile phase B in about three (3) minutes, and the final gradient is kept for another three (3) minutes. The anti-microbial agent, for example HP100, is detected at the negative MRM mode. The ion pair of m/z 253>142 is used as quantification transition, while m/z of 253>125 is used for identification.
In parallel, spiked matrix standards in the range of 0.5ng/ml to 500ng/ml are injected for creation of a calibration curve. Concentration of the anti-microbial agent, for example HP100, in the injection sample is determined by extrapolation using weighted (1/x ²) quadratic regression of the calibration curve.
Test 4: Biofilm Removal Test including biofilm generation (ASTM E2562) and removal in washing machines
1. Culture Preparation
Pseudomonas aeruginosa is the organism used in this test. Aseptically remove an isolated colony from an R2A plate and place into 100 mL of sterile TSB (300 mg/L) . Incubate bacterial suspension in an environmental shaker at 36 ± 2℃ for 22 ± 2 h. Viable bacterial density should equal 10 ⁸ CFU/mL, and may be checked by serial dilution and plating.
2. Reactor Preparation
A. Sonicate coupons in soap and tap water, rinse and sonicate the coupons with reagent grade water until no soap is left on the coupons.
B. Place a coupon into each hole in the reactor rods, tighten set screw. Place rods into reactor top loosely.
C. Invert the reactor top and place baffle onto glass rod positioned in the center of the reactor top.
D. Invert the reactor beaker and place onto the assembled top. Turn the reactor over so that the reactor top is upright.
E. Connect the bacterial air vent by fitting the vent to a small section of appropriately sized tubing, and attach to one of the rigid tubes on the reactor top.
F. The glass flow break is spliced into the nutrient tubing line near the reactor top.
G. Place the reactor top securely on the beaker before sterilization. To allow for pressure escape, do not set rod alignment pins in notches during sterilization.
H. Cover the end of the nutrient tubing that connects to the nutrient carboy and the end of the overflow (waste) tubing with aluminum foil. Cover any extra openings on the reactor top with aluminum foil. This is to maintain sterility after autoclaving.
I. Prepare batch culture medium by dissolving bacterial liquid growth medium (300 mg/L TSB) in 500 mL reagent grade water in an autoclavable container.
J. Sterilize the reactor system and separate batch culture medium for 20 min on the liquid cycle of a steam sterilizer.
3. Procedure
A. With the overflow (waste) line clamped, aseptically add the cooled batch culture medium to the cooled reactor.
B. Place reactor onto a stir plate.
C. Clamp flow break in upright position; leave other tubing clamped and foiled.
D. Secure the rod alignment pins into the reactor top notches.
E. Inoculate the reactor with 1 mL of bacteria from the culture prepared previously (see 2I) : Aseptically pipette the inoculum into the reactor through one of the available rigid reactor top tubes.
F. Turn on the magnetic stir plate. Set the rotational speed to 125 ± 5 r/min. The reactor system incubates in batch mode at room temperature (21 ± 2℃) for 24 h.
G. Prepare 100 mg/L TSB continuous flow nutrient broth. Dissolve and sterilize the broth in a smaller volume to prevent caramelization. Aseptically pour the concentrated broth into a carboy of sterile reagent grade water to make a total of 20 L.
H. Aseptically connect the nutrient tubing line to the carboy containing the continuous flow nutrient broth.
I. Pump a continuous flow of nutrients into the reactor at a flow rate determined by dividing the reactor volume by a 30 minute residence time. Attach tubing from the drain spout to a waste carboy and remove clamp. The drain spout on the beaker allows overflow to occur, maintaining a constant bacterial liquid growth broth concentration of 100 mg/L in the reactor during CSTR (continuously stirred tank reactor) mode.
J. Operate the reactor in SCTR mode for 24h.
4. Product Treatment
A. Turn off growth medium flow and baffled stir bar.
B. Aseptically remove a randomly selected rod containing coupons with biofilm from the CDC Biofilm Reactor by pulling it straight up and out of the reactor.
C. Rinse the coupons to remove planktonic cells: orient the rod in a vertical position directly over a 50 mL conical tube containing 30 mL sterile buffered water. With one continuous motion, immerse the rod into the buffered water with minimal to no splashing, then immediately remove. Use a new 50 mL conical tube with 30 mL sterile buffered water for each rod.
D. Prepare the product solutions in hardness water with recommended dosage (1000ppm) .
E. Place two rods into a customized beaker with top can hold the rods vertically. Transfer 350ml product solution in the beaker.
F. Put the beaker onto a magnetic stir plate and agitate for 10mins at 350rpm. Then discard the product solution and add 350ml fresh water to rinse the coupons for 3mins at 350rpm. Repeat the rinse once.
G. Apply 6-cycles product treatment for the coupons for each sample (step 4E-4F) .
H. For control, use 0.05%Tween80 to replace product solution and follow same treatments.
I. Remove an appropriate number of coupons for testing in individual tubes. Obtain a set of five coupons for each treatment and a set of three coupons for the controls.
J. Add 3ml PBS solution into each tube containing one treated coupon.
K. Vortex each tube on the highest setting, ensuring a complete vortex for 30 ± 5 s.
L. Sonicate the tubes at 45 ± 5 kHz for 30 ± 5 s at room temperature (21 ± 2 ℃) (use normal mode if sonicator has variable settings) .
M. Vortex each tube on the highest setting, ensuring a complete vortex for 30 ± 5 s.
N. Sonicate the tubes at 45 ± 5 kHz for 30 ± 5 s at room temperature (21 ± 2 ℃) (use normal mode if sonicator has variable settings) .
O. Vortex each tube on the highest setting, ensuring a complete vortex for 30 ± 5 s. These tubes are the 10 ⁰ dilution.
P. Dilute and count bacteria amount on each coupon, transfer the value to log ₁₀ and take the average of control coupons and product treated coupons.
5. Calculation of biofilm removal efficacy
Log Reduction = average of log ₁₀ (control coupons) –average of log ₁₀ (product treated coupons) .
EXAMPLES
The Examples herein are meant to exemplify the present invention but are not used to limit or otherwise define the scope of the present invention.
Example 1: Comparative Test Showing Effective Microbial Removal by Anti-microbial Composition Containing 4-chloro-3, 5-dimethyl phenol and a Hydroxyl Diphenyl Ether
Four (4) sample liquid compositions were prepared containing the ingredients as shown in Table 1 below, in which Samples 1 to 3 comprise 4-chloro-3, 5-dimethyl phenol (PCMX) as an active for microbial removal and a hydroxyl diphenyl ether as an active for microbial prevention, and Comparative Sample 1 comprises a hydroxyl diphenyl ether, but no PCMX.
Table 1
a 4-chloro-3, 5-dimethyl phenol available from Jiangsu Huanxin High-Tech materials Co., Ltd
b HP100 is 4-4’-dichloro-2-hydroxy diphenyl ether, available from BASF
c available from SENYU FORREST Chemical GuangDong
d available from Chenxin chemical, Guangzhou. Ltd.
e available from SHANDONG SHIDA SHENGHUA CHEMICAL
Then, the efficacy of microbial removal for the above samples at a dose of 1: 50 (i.e., 2%finished product in water) was determined in accordance with Test 1: Efficacy of Microbial Removal in which Gram-negative bacteria K. pneumoniae was used. The results are shown below.
Table 2
As the data shown above, the samples comprising both PCMX and a hydroxyl diphenyl ether can deliver a microbial removal efficacy. Particularly, when the TTW level of PCMX is at least 100ppm, the efficacy is very strong (i.e., up to 99.99%) .
Example 2: Comparative Test Showing Effective Microbial Prevention by Anti-microbial Composition Containing 4-chloro-3, 5-dimethyl phenol and a Hydroxyl Diphenyl Ether
Twelve (12) sample liquid compositions were prepared containing the ingredients as shown in Table 1 below, in which Samples 1 to 8 comprise 4-chloro-3, 5-dimethyl phenol (PCMX) as an active for microbial removal and a hydroxyl diphenyl ether as an active for microbial prevention, and Comparative Samples 1 to 4 comprises only one of PCMX and a hydroxyl diphenyl ether.
Table 3a
a 4-chloro-3, 5-dimethyl phenol available from Jiangsu Huanxin High-Tech materials Co., Ltd
b is 4-4’-dichloro-2-hydroxy diphenyl ether, available from BASF
c available from SENYU FORREST Chemical GuangDong
d available from Chenxin chemical, Guangzhou. Ltd.
e available from SHANDONG SHIDA SHENGHUA CHEMICAL
Table 3b
a 4-chloro-3, 5-dimethyl phenol available from Jiangsu Huanxin High-Tech materials Co., Ltd
b is 4-4’-dichloro-2-hydroxy diphenyl ether, available from BASF
c available from SENYU FORREST Chemical GuangDong
d available from Chenxin chemical, Guangzhou. Ltd.
e available from SHANDONG SHIDA SHENGHUA CHEMICAL
Then, the efficacy of microbial prevention for the above samples at a dose of 1: 50 (i.e., 2%finished product in water) was determined in accordance with Test 2: Efficacy of Microbial Prevention in which Gram-negative bacteria K. pneumoniae was used. The results are shown below.
Table 4a
Table 4b
Surprisingly, when the TTW dosage of PCMX in the samples is not too high e.g. lower than 1000 ppm, the samples containing both PCMX and the hydroxyl diphenyl ether can effectively achieve microbial prevention (i.e., logR is at least 2) . In view of Example 1, these samples can deliver both the efficacies of microbial removal and microbial prevention. In the contrast, if the TTW dosage of PCMX is too high, e.g. 1000 ppm, the efficacy provided by the hydroxyl diphenyl ether would be significantly compromised (i.e., only 0.6) .
Accordingly, in order to achieve both microbial removal and microbial prevention, a preferred range of TTW dosage of PCMX is needed, e.g. from 20 ppm to 900 ppm, preferably from 40 ppm to 800 ppm.
Example 3: Comparative Test Showing Improved Deposition of Anti-microbial Agent on Fabrics by Anti-microbial Composition Containing 4-chloro-3, 5-dimethyl phenol and a Hydroxyl Diphenyl Ether
Samples 1 to 6 and Comparative Sample 1 were prepared similarly as in Examples 1 and 2, in which Samples 1 to 6 comprise 4-chloro-3, 5-dimethyl phenol (PCMX) as an active for microbial removal and a hydroxyl diphenyl ether as an active for microbial prevention, and Comparative Sample 1 comprises a hydroxyl diphenyl ether without PCMX.
Table 5
a 4-chloro-3, 5-dimethyl phenol available from Jiangsu Huanxin High-Tech materials Co., Ltd
b is 4-4’-dichloro-2-hydroxy diphenyl ether, available from BASF
c available from SENYU FORREST Chemical GuangDong
d available from Chenxin chemical, Guangzhou. Ltd.
e available from SHANDONG SHIDA SHENGHUA CHEMICAL
The deposition of the hydroxyl diphenyl ether on fabrics for these samples were determined in accordance with Test 3: Fabric Deposition Test for Anti-Microbial Agents. The results are shown below:
Table 6
It is greatly surprising from the data as shown above that, when the TTW dosage of PCMX in the formulation containing PCMX and a hydroxyl diphenyl ether is within the preferred range, e.g. 40 ppm to 500 ppm in Samples 1 to 5, the deposition of the hydroxyl diphenyl ether is significantly improved (3.31-8.2 vs. 1.93) , which is completely unexpected. On the contrary, when the TTW dosage of PCMX is 1000 ppm, the deposition of hydroxyl diphenyl ether is almost completely blocked (only 0.20) .
Example 4: Exemplary Formulations of Compositions
The following liquid compositions shown in Table 7 are made comprising the listed ingredients in the listed proportions (weight %) .
Table 7
a 4-chloro-3, 5-dimethyl phenol available from Jiangsu Huanxin High-Tech materials Co., Ltd
b is 4-4’-dichloro-2-hydroxy diphenyl ether, available from BASF
c available from SENYU FORREST Chemical GuangDong
d available from Chenxin chemical, Guangzhou. Ltd.
e available from SHANDONG SHIDA SHENGHUA CHEMICAL
f Linear C12-C14 alkoxylated (EO7) alcohol
g C10-C14 linear alkyl benzene sulfonate
h Sodium Laureth 3 Sulfate
The dimensions and values disclosed herein are not to be understood as being strictly limited to the exact numerical values recited. Instead, unless otherwise specified, each such dimension is intended to mean both the recited value and a functionally equivalent range surrounding that value. For example, a dimension disclosed as “40 mm” is intended to mean “about 40 mm. ”
Every document cited herein, including any cross referenced or related patent or application and any patent application or patent to which this application claims priority or benefit thereof, is hereby incorporated herein by reference in its entirety unless expressly excluded or otherwise limited. The citation of any document is not an admission that it is prior art with respect to any invention disclosed or claimed herein or that it alone, or in any combination with any other reference or references, teaches, suggests or discloses any such invention. Further, to the extent that any meaning or definition of a term in this document conflicts with any meaning or definition of the same term in a document incorporated by reference, the meaning or definition assigned to that term in this document shall govern.
While particular embodiments of the present invention have been illustrated and described, it would be obvious to those skilled in the art that various other changes and modifications can be made without departing from the spirit and scope of the invention. It is therefore intended to cover in the appended claims all such changes and modifications that are within the scope of this invention.

Claims

An anti-microbial composition comprising:

a) 4-chloro-3, 5-dimethyl phenol; and

b) hydroxyl diphenyl ether of formula (I) :

wherein:

each Y is independently selected from chlorine, bromine, or fluorine,

each Z is independently selected from SO ₂H, NO ₂, or C ₁-C ₄ alkyl,

r is 0, 1, 2, or 3,

o is 0, 1, 2, or 3,

p is 0, 1, or 2,

m is 1 or 2, and

n is 0 or 1;

wherein 4-chloro-3, 5-dimethyl phenol is present in an amount sufficient for delivering a Through-The-Wash (TTW) dosage ranging from 20 ppm to 960 ppm, preferably from 40 ppm to 800 ppm, more preferably from 80 ppm to 400 ppm, yet more preferably from 120 ppm to 300 ppm, most preferably from 160 ppm to 240 ppm.
The composition according to any one of the preceding claims, wherein said hydroxyl diphenyl ether is present in an amount sufficient for delivering a Through-The-Wash (TTW) dosage ranging from 0.1 ppm to 400 ppm, preferably from 0.5 ppm to 100 ppm, more preferably from 1 ppm to 20 ppm, yet more preferably from 1.5 ppm to 12 ppm, most preferably from 2 ppm to 8 ppm.
The composition according to any one of the preceding claims, wherein 4-chloro-3, 5-dimethyl phenol is present in an amount ranging from 0.1%to 4.8%, preferably from 0.2%to 4%, more preferably from 0.4%to 2%, yet more preferably from 0.6%to 1.5%, most preferably from 0.8%to 1.2%, by weight of the composition; and/or

wherein said hydroxyl diphenyl ether is present in an amount ranging from 0.0005%to 2%, preferably from 0.0025%to 0.5%, more preferably from 0.005%to 0.1%, yet more preferably from 0.0075%to 0.06%, most preferably from 0.01%to 0.04%, by weight of the composition.
The composition according to any one of the preceding claims, , wherein the weight ratio of 4-chloro-3, 5-dimethyl phenol to said hydroxyl diphenyl ether is between 0.1 and 2000, preferably between 1 and 1000, more preferably between 5 and 200, yet more preferably between 30 and 70, yet more preferably between 35 and 65, yet more preferably between 40 and 60, and most preferably between 45 and 55.
The composition according to any one of the preceding claims, wherein said hydroxyl diphenyl ether is selected from the group consisting of 4-4’ -dichloro-2-hydroxy diphenyl ether, 2, 4, 4’ -trichloro-2’ -hydroxy diphenyl ether, and a combination thereof, preferably 4-4’ -dichloro-2-hydroxy diphenyl ether.
The composition according to any one of the preceding claims, wherein said composition further comprises from 1%to 50%, more preferably from 3%to 40%, most preferably from 5%to 30%, by total weight of said composition, of a solvent;

preferably, wherein said solvent is an alcohol selected from the group consisting of ethanol, ethylene glycol, propylene glycol, glycerol, butanediol, butanetriol, poly (propylene glycol) , terpineol, and a combination thereof;

more preferably, wherein said solvent is propylene glycol and/or terpineol.
The composition according to any one of the preceding claims, wherein said composition further comprises from 0.1%to 30%, more preferably from 0.5%to 20%, most preferably from 1%to 10%, by total weight of said composition, of a crystalline, hydroxyl-containing stabilizing agent;

preferably, wherein said stabilizing agent is selected from the group consisting of microcrystalline cellulose or derivatives thereof, castor oil or derivatives thereof, hydrogenated castor oil or derivatives thereof, and any combinations thereof;

more preferably, wherein said stabilizing agent is castor oil.
The composition according to any one of the preceding claims, wherein said composition further comprises from 0.01%to 5%, preferably from 0.02%to 2%, more preferably from 0.05%to 1%, of a perfume by total weight of said composition; wherein said perfume is preferably selected from the group consisting of free perfumes, pro-perfumes, encapsulated perfumes, perfume microcapsules, and combinations thereof; wherein said perfume more preferably comprises a combination of free perfumes and perfume microcapsules; wherein most preferably the weight ratio of free perfumes to perfume microcapsules in said composition is from 1: 5 to 20: 1, preferalby from 1: 2 to 10: 1, more preferably from 1: 1 to 5: 1, most preferably from 1.5: 1 to 3: 1.
The composition according to any one of the preceding claims, wherein said composition is a liquid composition, preferably a liquid sanitizer.
A process for treating an article of clothing comprising the steps of:

a) providing an article of clothing and an anti-microbial composition according to any one of the preceding claims, and

b) contacting said article of clothing with said composition.