Abstract:
A mascara applicator brush is provided, including polymer bristles which are coated with a germicidal composition. The aforementioned composition is made from a mixture based on at least one large cation and at least one large anion, in which one or both develop germicidal properties.

Description:
FIELD OF THE INVENTION 
     The invention relates to a brush having germicidal properties for applying mascara. 
     BACKGROUND OF THE INVENTION 
     Mascara is a cosmetic product used to color and/or thicken eyelashes. This composition is applied to the eyelashes with a brush that is traditionally attached to the plug that seals the bottle containing the mascara composition. In doing this, once the bottle is first opened, successive applications of the mascara with this brush and reinserting the brush into the bottle containing the mascara itself thus leads to a risk of contamination of the cosmetic composition with germs, and notably bacteria or fungi (i.e., germs picked up by the brush when in contact with the user&#39;s skin or eyelashes). These germs enter into the cosmetic composition, thus leading to a relatively fast termination of the mascara&#39;s usability, in fact requiring that it should be discarded relatively quickly in order to limit the possible risks of the multiplication of these germs in the mascara, in any case well before the cosmetic composition is used up. 
     To remedy this disadvantage, it has long been suggested that the mascara composition should contain preservatives, i.e. additives which can avoid and at least limit the growth of these germs. It has been shown, however, that such preservatives are irritants and can cause reactions in the eye, very close to the area where the mascara is applied. Moreover, the use of such preservatives can lead to the creation of resistant bacterial strains which can develop in the eye, risking the accompanying pathological consequences. 
     In a related field, concerning oral hygiene, the proposal has been made, notably in document WO 99/35911, to produce a toothbrush whose bristles are made of a plastic material containing a compound with antimicrobial activity, the component notably comprising a halogenated hydrocarbon, notably triclosan. 
     Experience has shown, however, that while germicidal activity can indeed be demonstrated, the salting out of this antimicrobial compound is observed, notably in the oral cavity. While such salting out has no effects, or at least no harmful effects, in toothbrush applications, it is unacceptable in the case of mascara, where such salting out would occur in the cosmetic composition itself, which could affect its composition and, furthermore, would not provide any increase in the duration of use of the mascara applicator brush. 
     SUMMARY OF THE INVENTION 
     Indeed, the object of the present invention is, on the one hand, to increase the useful lifetime of the mascara applicator brush, and therefore to optimize the use-by date for such a product. The present invention also aims to decrease, as much as possible, the quantity of preservatives included in the cosmetic composition, avoiding the salting out of the germicide in the mascara as much as possible. 
     In the following description and claims, the term “large anion” refers to an anion selected from the group consisting of:
         anions of the carboxylate (oleate, for example) or alkyl sulphate (lauiyl sulphate, for example) type, with an alkyl chain having a number of carbons greater than 10;   polyanions of the polycarboxylate type (polyacrylate, for example); or   other anions of the silicate or polyphosphate type.       

     In the following description and claims, the term “large cations” refers to a cation selected from the group consisting of:
         quaternary ammoniums bearing at least one alkyl chain having a number of carbons greater than 8;   cationic polymers of the ammonium polyacrylate type;       

     polyiminium hydrochlorides and notably polyhexamethylene biguanide (PHMB); and
         polymers bearing quaternary ammonium functions and notably quaternary polyammoniums.       

     The present invention provides a mascara applicator brush comprising polymer bristles coated with a germicidal composition produced using a mixture
         including at least one large cation and at least one large anion, one or the other or both developing germicidal properties.       

     Implementation of this particular mixture thus allows a compound with germicidal properties to bind to the bristles on the brush, experience showing that it is not salted out into the mascara. At the same time, this compound provides the satisfactory development of germicidal properties and in all cases in compliance with the goal sought by the present invention. 
     Advantageously, the polymer constituting the bristles of the brush is a polyamide, and preferably polyamide 6.12. 
     It is possible, however, to envisage implementing a synthetic polymer chosen from the group including polyurethane, polyethylene, polypropylene, polyester, polyacrylic, modacrylic, alone or in mixtures. 
     The polymer constituting the bristles of the brush can also be an artificial or natural polymer. 
     According to another aspect of the invention, the cation implemented comes from a polyiminium salt (hydrochloride, for example), and notably polyhexamethylene biguanide, more commonly known as PHMB. This cation can also be made of a quaternary ammonium salt, notably quaternary polyammonium. 
     In one advantageous production method, the large anion is derived from a sodium polyacrylate salt, sodium silicate, sodium polyphosphate, sodium oleate or sodium lauryl sulphate. 
     The Applicant has observed that particularly interesting results can be obtained in terms of germicidal properties and the absence of salting out into the mascara when the composition of the invention combines sodium oleate and PHMB, advantageously in 50/50 mole proportions. 
     The present invention also relates to a method for depositing such a germicidal composition onto the mascara applicator brushes. 
     This method includes:
         performing cold soaking of the brush in a mixture of at least one large cation salt and at least one large anion salt, one or the other or both having germicidal properties;   then performing a rinsing step to eliminate excess substance not bound to the bristles of the brush; and   then performing a drying step to eliminate the water contained in the brushes.       

     In other production methods, the anion and cation salts can be mixed successively, one before the other and vice-versa. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIGS. 1 and 2  represent an illustrative graph of the germicidal action of the composition of the invention against  Escherichia coli  CIP 53.126 under normal conditions of use. 
         FIG. 3  represents an illustrative graph of the germicidal action of the composition of the invention against  Staphylococcus aureus  CIP 4.83 under normal conditions of use. 
     
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     The present invention and its advantages can be seen in the following examples, and are supported by the appended drawing figures. 
     Example 1 
     Preparation of Brushes with Germicidal Properties 
     5300 brushes, produced using polyamide 6.12 bristles with a diameter of 80 micrometers (representing a weight of 1500 grams) are placed in 10 litres and treated as follows:
         Prepare 4.5 kg of a solution of sodium oleate at 0.69 wt. %: dissolve 31.05 g pure sodium oleate in approximately 500 ml warm soft water, fill to 4.5 kg with cold soft water.   Prepare 4.5 kg of a solution of Cosmocil CQ (PHMB sold by Avecia at 20% by weight in water) at 0.5% by weight: dissolve 112.5 g Cosmocil CQ (at 20% in water) in 500 ml cold soft water; fill to 4.5 kg with cold soft water.   Add the 4.5 kg of Cosmocil CQ solution at 0.5% into the reactor (total volume of liquid: 9 litres for a bath ratio of 6).   Stir the bath.   Add the 4.5 kg of sodium oleate solution at 0.69% into the reactor while stirring.   Continue stirring.       

     The brushes are removed from the reactor, then rinsed in soft water and dried. 
     The weight increase of the brushes is 1.84%. 
     Example 2 
     Microbiological Results of the Brush Treated According to the Invention with Mascara Absent 
     The purpose of this example is to test the germicidal properties of the composition of the invention when applied according to the method in example 1 to various types of fibres constituting a mascara brush. 
     The composition tested is the following:
         COSMOCIL CQ® bactericide manufactured by AVECIA in a 20% solution: —((CH 2 ) 3 —NH—CNH—NH—CNH—NH—(CH 2 ) 3 C) n —HCl, n=16   anion: SODIUM OLEATE manufactured by RIEDEL DEHAEN: CH 3 (CH 2 ) 7 CH═CH(CH 2 ) 7 CO 2 NaC 17 H 33 CO 2 Na=304.4       

     The composition of the invention is applied to three different types of brushes, respectively: 
     1. polyamide 6.12 
     2. polyamide 6.6 
     3. polyamide 6 (two origins) 
     In practice, each brush sample is immersed in the COSMOCIL CQ/SODIUM OLEATE solution stirred for several hours at ordinary temperature. The bath ratio, liquid mass/brush mass, is between 3 and 10 (more advantageously, 6). The brushes are rinsed, centrifuged and dried. 
     For samples corresponding solely to the STRAND 14 and STRAND 14R references in polyamide 6.12, the brushes are previously rinsed in hot water and dried at 80° C. 
     The treated brushes are, depending on the case, post-treated by rinsing with soft water, whether cold or not. 
     The dry matter content of each tested sample is represented in the following table: 
     
       
         
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 1 
               
             
             
               
                   
               
               
                 Results 
               
             
          
           
               
                   
                   
                 Pre- 
                 Molar 
                 Post- 
                 Dry matter 
               
               
                 Samples 
                 Support 
                 treatment 
                 ratio 
                 treatment 
                 content 
               
               
                   
               
               
                 Strand 14 
                 PA 6.12 
                 Rinse with 
                 1/1 
                   
                 2.04% 
               
               
                 Strand 
                   
                 water and dry 
                 1/1 
                 rinsed 
                 2.04% 
               
               
                 14R 
                   
                 at 80° C. 
               
               
                 A2 
                 PA 6.12 
                   
                 1/1 
                   
                 4.46% 
               
               
                 B2 
                 PA 6.6 
                   
                 1/1 
                   
                 5.02% 
               
               
                 C2 
                 PA 6 
                   
                 1/1 
                   
                 4.93% 
               
               
                   
                 Ref. A 
               
               
                 D2 
                 PA 6 
                   
                 1/1 
                   
                 3.96% 
               
               
                   
                 Ref. B 
               
               
                 A2R 
                 PA 6.12 
                   
                 1/1 
                 rinsed 
                 3.42% 
               
               
                 B2R 
                 PA 6.6 
                   
                 1/1 
                 rinsed 
                 3.11% 
               
               
                 C2R 
                 PA 6 
                   
                 1/1 
                 rinsed 
                 3.05% 
               
               
                   
                 Ref. A 
               
               
                 D2R 
                 PA 6 
                   
                 1/1 
                 rinsed 
                 6.36% 
               
               
                   
                 Ref. B 
               
               
                   
               
             
          
         
       
     
     The following microbiological test was then performed on each sample: 
     The fibres are placed in suspension in the peptone broth contaminated with five different germs:
           Escherichia coli  CIP 53.126 (for STRAND 14 and STRAND 14R only)     Staphylococcus aureus  CIP 4.83 (except for STRAND 14 and STRAND 14R samples)     Pseudomonas aeruginosa  CIP 82.118 (except for STRAND 14 and STRAND 14R samples)     Candida albicans  IP 48.72 (except for STRAND 14 and STRAND 14R samples)     Aspergillus niger  IP 1431.33 (except for STRAND 14 and STRAND 14R samples)       

     The evolution of contamination is measured for the bacteria every day for one week and at D+1, D+5 and D+7 for yeasts and moulds. Values are given in CFU/ml. Each test is performed in triplicate. 
     The results are given in tables 2 to 6 below. 
     
       
         
               
             
               
               
               
             
           
               
                 TABLE 2 
               
             
             
               
                   
               
               
                   Escherichia coli  CIP 53.126 
               
             
          
           
               
                 Time 
                 STRAND 14 
                 STRAND 14R 
               
               
                   
               
               
                 D0 
                 1.67 × 10 5   
                 1.60 × 10 5   
               
               
                 D + 1 
                 &lt;2 
                 &lt;2 
               
               
                 D + 2 
                 &lt;2 
                 &lt;2 
               
               
                 D + 5 
                 &lt;2 
                 &lt;2 
               
               
                 D + 6 
                 &lt;2 
                 &lt;2 
               
               
                 D + 7 
                 &lt;2 
                 &lt;2 
               
               
                 D + 8 
                 &lt;2 
                 &lt;2 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 3 
               
             
             
               
                   
               
               
                   Staphylococcus aureus  CIP 4.83 
               
             
          
           
               
                 Time 
                 A2 
                 A2R 
                 B2 
                 B2R 
                 C2 
                 C2R 
                 D2 
                 D2R 
               
               
                   
               
               
                 D0 
                 2.8 × 
                 3.0 × 
                 4.0 × 
                 3.7 × 
                 7.0 × 
                 6.0 × 
                 5.1 × 
                 8.4 × 
               
               
                   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
               
               
                 D + 1 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
               
               
                 D + 2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
               
               
                 D + 5 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
               
               
                 D + 6 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 4 
               
             
             
               
                   
               
               
                   Pseudomonas aeruginosa  CIP 82.118 
               
             
          
           
               
                 Time 
                 A2 
                 A2R 
                 B2 
                 B2R 
                 C2 
                 C2R 
                 D2 
                 D2R 
               
               
                   
               
               
                 D0 
                 2.8 × 
                 2.4 × 
                 3.2 × 
                 3.3 × 
                 5.0 × 
                 5.7 × 
                 4.6 × 
                 7.9 × 
               
               
                   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
               
               
                 D + 1 
                 &lt;2 
                 2.2 × 
                 &lt;2 
                 9.5 × 
                 3.0 × 
                 2.6 × 
                 &lt;2 
                 13 
               
               
                   
                   
                 10 2   
                   
                 10 2   
                 10 2   
                 10 2   
               
               
                 D + 2 
                 &lt;2 
                 2.4 × 
                 &lt;2 
                 1.6 × 
                 1.8 × 
                 1.0 × 
                 &lt;2 
                 4.4 × 
               
               
                   
                   
                 10 3   
                   
                 10 3   
                 10 3   
                 10 3   
                   
                 10 2   
               
               
                 D + 5 
                 &lt;2 
                 2.5 × 
                 3.7 × 
                 6.4 × 
                 3.8 × 
                 1.9 × 
                 &lt;2 
                 9.5 × 
               
               
                   
                   
                 10 2   
                 10 2   
                 10 3   
                 10 5   
                 10 4   
                   
                 10 3   
               
               
                 D + 6 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 5 
               
             
             
               
                   
               
               
                   Candida albicans  IP 48.72 
               
             
          
           
               
                 Time 
                 A2 
                 A2R 
                 B2 
                 B2R 
                 C2 
                 C2R 
                 D2 
                 D2R 
               
               
                   
               
               
                 D0 
                 2.8 × 
                 4.1 × 
                 5.1 × 
                 4.7 × 
                 4.2 × 
                 4.5 × 
                 4.5 × 
                 4.4 × 
               
               
                   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
               
               
                 D + 1 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
               
               
                 D + 5 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
               
               
                 D + 6 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 6 
               
             
             
               
                   
               
               
                   Aspergillus niger  IP 1431.33 
               
             
          
           
               
                 Time 
                 A2 
                 A2R 
                 B2 
                 B2R 
                 C2 
                 C2R 
                 D2 
                 D2R 
               
               
                   
               
               
                 D0 
                 1.6 × 
                 1.5 × 
                 2.1 × 
                 3.8 × 
                 1.7 × 
                 2.6 × 
                 4.0 × 
                 2.2 × 
               
               
                   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
                 10 4   
               
               
                 D + 1 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
               
               
                 D + 5 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
               
               
                 D + 6 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
                 &lt;2 
               
               
                   
               
             
          
         
       
     
     We observe that the STRAND 14 (not rinsed) and STRAND 14R (rinsed) samples are effective against  Escherichia coli  CIP 53.126. 
     We also observe that samples A2, B2, C2 and D2 present good antibacterial and antifungal activity. Indeed, we observe a rapid decrease: count lower than 2 CFU/ml in 24 hours. 
     Against the  Pseudomonas aeruginosa  CIP 82.118 strain, samples A2 and D2 present good antibacterial activity, since a rapid decrease is observed to a threshold under 2 CFU in 24 hours. Likewise, samples B2 and C2 present good antibacterial activity in 6 days, since the decrease reaches a threshold under 2 CPU/ml. 
     Measurement of the Salting Out of the Germicidal Composition into the Mascara 
     This measurement of salting out into the mascara is performed under normal conditions of use, i.e. at the level of the actual cosmetic compound contained in the bottle. It consisted in quantifying or detecting the bactericidal matter, COSMOCIL®, bound to the bristles of the brush according to the method previously described. 
     For this, the brushes are placed in contact continuously for 8 days at ambient temperature and at 40° C. Negative controls and COSMOCIL® (20% solution) are used to calibrate the measurement device used. 
     The various measurements made show that, in all cases, detection is below 0.003%. 
     Example 3 
     Measurement of the Antimicrobial Activity of PURCILON® Fibres Mounted on a Brush Under Real Conditions of Use 
     In this example, we verify the antimicrobial activity of fibres treated with PURCILON® mounted on a brush under real conditions of use, i.e. for mascara. 
     Composition of PURCILON®: Polyamide 6.12 with 4.7% dry material content (similar to the A2R reference) treated according to the method described above on 1000 brushes. 
     Material and Method 
     Mascara Base 
     The base formula chosen is black water-resistant mascara with the following preservative system: 
     
       
         
               
               
               
             
           
               
                   
                   
               
             
             
               
                   
                 Ethyl para-hydroxybenzoate (E POB) 
                 0.20% 
               
               
                   
                 Methyl para-hydroxybenzoate (M POB) 
                 0.10% 
               
               
                   
                 Propyl para-hydroxybenzoate (P POB) 
                 0.155%  
               
               
                   
                 Benzyl alcohol, methyl-4-hydroxybenzoate, 
                 0.50% 
               
               
                   
                 propyl-4-hydroxybenzoate 
               
               
                   
                   
               
             
          
         
       
     
     This base also contains matter that can facilitate the action of the preservatives such as: 
     
       
         
               
               
               
             
           
               
                   
                   
               
             
             
               
                   
                 Tetrasodium salt of 
                 0.10%   
               
               
                   
                 ethylenediaminetetraacetic acid (EDTA) 
               
               
                   
                 Glycerine, water, 1,2-octanediol, PEG-8, 
                 3% 
               
               
                   
                 sodium polyacrylate 
               
               
                   
                 Butylene glycol 
                 1% 
               
               
                   
                   
               
             
          
         
       
     
     The preservative was validated according to criteria B of the European pharmacopoeia. 
     Strains 
     The protective power of the mascara was studied for the following microbial strains:
           Escherichia coli  CIP 53.126     Staphylococcus aureus  CIP 4.83       

     The strains are maintained by deep freezing. They are used after Trypcase soy agar subculturing. 
     Experimental Protocol 
     Controls and Trials Performed 
     The trials performed with the treated fibres and mascara with preservatives are called Ft trials. 
     At the same time, controls are made using the same protocol:
         Ta: treated, non-contaminated fibres placed in contact with mascara without preservatives, which is used to verify the cleanliness of the treated fibres as well as that of the small bottles.   Tb: non-treated, non-contaminated fibres placed in contact with mascara without preservatives, which is used to verify the cleanliness of the non-treated fibres as well as that of the small bottles.   Tc: treated, contaminated fibres placed in contact with mascara without preservatives, which is used to verify the real incidence of the bactericidal effect of the treated fibres.   Td: non-treated, contaminated fibres placed in contact with mascara containing preservatives, which is used to verify the incidence of the preservative system in the mascara on decreasing the germ concentration.   Te: non-treated, contaminated fibres placed in contact with mascara without preservatives.       

     The principle consists in contaminating the brushes by soaking them in a germ solution and then inserting them into the small mascara bottles containing 6 grams of the Cilpur® formula and then monitoring the evolution of the contamination over time. 
     Each stock solution of germs is placed in the empty small bottles which are previously decontaminated with gamma rays. The diaphragm in the bottle provides a good calibration of the volume retained on the brush (estimated volume: 0.0708 ml over 10 trials). 
     The protective power of the fibres was studied using:
         two stock solutions of  E. coli  calibrated to obtain an initial contamination of approximately 10 8  CFU/ml for S1 and 10 7  CFU/ml for S2.   one stock solution of  S. aureus  calibrated to obtain an initial contamination of approximately 10 5  CFU/ml.       

     To evaluate the initial quantity of germs, the brush is used to retrieve:
         for  E. coli,  0.5 g±0.05 g mascara in 9 ml Eugon LT100 (neutralising diluent).   for  S. aureus,  0.25 g±0.01 g mascara in 9 ml Eugon LT100.       

     Then, 0.5 ml of each sample is inoculated. The agars are then incubated at 30-35° C. The small bottles are sealed with the brushes and stored at 20-25° C. 
     Checks Performed on  E. coli:    
     The bottles are checked after 24 hours of contact time for the first contamination. Two other overcontaminations are then performed on the same mascara bottles with the same brushes and checks on the evolution of contaminations are performed after 6 hours and 24 hours of contact. 
     Checks Performed on  S. aureus:    
     The bottles are checked after 1, 2 and 6 hours of contact for the first two contaminations. For the third overcontamination, the checks are performed after 1, 2, 6 and 24 hours of contact. 
     All trials and controls are performed in triplicate as are the agar inoculation, which makes it possible to perform a statistical assessment of the results and to eliminate abnormal values. 
     Results and Discussion 
     Populations are determined using the results of viable germ counts. After eliminating the abnormal values, an average of the various trials is calculated. 
     
       Escherichia coli:  
     
     The trials performed with solutions S1 and S2 are fairly similar as can be seen in  FIGS. 1 and 2 . 
     A sharp decrease in germs is observed for trials Td and Ft (approximately 2 log in 6 hours). Comparison with the results obtained for trials Tc and Te can be used to determine that this log reduction is directly linked to the action of the preservatives present in the mascara. 
     Trials Td and Ft, notably with solution S2, demonstrate an improvement in the log reduction when the action of the preservatives in the mascara is combined with those present in the treated fibres. The smaller the initial population of viable germs, the greater this improvement. Trials Tc and Te back up this hypothesis of a synergistic action between the preservatives present in the mascara and in the treated fibres. 
     
       Staphylococcus aureus:  
     
     As was the case for  E. coli , a sharp decrease in germs is observed for trials Td and Ft ( FIG. 3 ). Comparison with the results obtained for trials Tc and Te can be used to determine that this log reduction is directly linked to the combined action of the preservatives present in the mascara and the germicide present in the treated fibres.