Process for preparing an antimicrobial particulate composition

Abstract

The present invention relates to a process for preparing an antimicrobial particulate composition. The invention also relates to personal care or hygiene composition comprising an antimicrobial particulate composition obtainable by the process in accordance with this invention. The invention more particularly relates to a process for preparing an antimicrobial metal nanoparticles immobilized in an inorganic porous material and also incorporating these particles in personal care or hygiene compositions. The antimicrobial particulate composition comprises 0.05% to 3% by weight of antimicrobial metal particles and 97 to 99.95% by weight of immobilizer comprising an inorganic porous material selected from zinc oxide, magnesium hydroxide or calcium carbonate.

Claims

1. A process for preparing an antimicrobial particulate composition the process comprising the steps of: i. mixing an aqueous dispersion of an immobilizer, wherein the immobilizer is present in an amount of 1-5 wt % (by weight of the dispersion) and wherein the immobilizer is selected from inorganic porous material selected from zinc oxide, magnesium hydroxide or calcium carbonate and wherein the immobilizer has a particle size in the range of 1-10 microns and an aqueous solution of a reducing agent, wherein the reducing agent is present in the aqueous solution in an amount of from 10 to 30 wt % (by weight of the immobilizer in the aqueous dispersion); ii. raising the temperature of the mixture resulting from step (i) to a temperature in the range 70 C. to 90 C.; iii. adding a water soluble metal salt to the mixture resulting from step (ii) in an amount equivalent to 0.05 to 3% of the metal by weight of the immobilizer under mixing.

2. The process according to claim 1, wherein the water soluble metal salt is selected from water soluble salts of silver or of copper.

3. The process according to claim 1, wherein the water soluble metal salt is a water soluble salt of silver.

4. The process according to claim 3, wherein the water soluble salt of silver is selected from silver nitrate or silver acetate.

5. The process according to claim 2, wherein the water soluble salt of copper is selected from copper (II) sulphate, copper (II) nitrate, copper (II) chloride, or copper (II) acetate.

6. The process according to claim 1, wherein the reducing agent is selected from a water soluble salt of a carboxylic acid, wherein the reducing agent has 1 to 4 carboxylate groups.

7. The process according to claim 1, wherein the reducing agent is selected from sodium acetate, sodium oxalate, trisodium citrate or disodium ethylene diamine tetra acetate.

Description

DETAILED DESCRIPTION OF THE INVENTION

(1) These and other aspects, features and advantages will become apparent to those of ordinary skill in the art from a reading of the following detailed description and the appended claims. For the avoidance of doubt, any feature of one aspect of the present invention may be utilized in any other aspect of the invention. The word comprising is intended to mean including but not necessarily consisting of or composed of. In other words, the listed steps or options need not be exhaustive. It is noted that the examples given in the description below are intended to clarify the invention and are not intended to limit the invention to those examples per se. Similarly, all percentages are weight/weight percentages unless otherwise indicated. Except in the operating and comparative examples, or where otherwise explicitly indicated, all numbers in this description and claims indicating amounts of material or conditions of reaction, physical properties of materials and/or use are to be understood as modified by the word about. Numerical ranges expressed in the format from x to y are understood to include x and y. When for a specific feature multiple preferred ranges are described in the format from x to y, it is understood that all ranges combining the different endpoints are also contemplated.

(2) The antimicrobial particulate composition made in accordance with the invention comprises 0.05% to 3% by weight of antimicrobial metal particles and 97 to 99.95% by weight of immobilizer comprising inorganic porous material selected from zinc oxide, magnesium hydroxide or calcium carbonate.

(3) The weight % of the antimicrobial metal particles and the immobilizer in the antimicrobial particulate composition is by weight of the composition.

(4) Antimicrobial Metal Particles:

(5) It is preferred that the antimicrobial metal particles are preferably selected from silver or copper particles, more preferably from silver or copper nanoparticles and most preferably are silver nanoparticles.

(6) Immobilizer:

(7) The immobilizer is selected from an inorganic porous material selected from porous zinc oxide, magnesium hydroxide or calcium carbonate and it is preferred that the inorganic porous materials have a nano/micro-structured assembly and more preferably are aggregates of nano plate like structures having preferably 20-100 nm width and more preferably 40-60 nm width. It is preferred that the immobilizer has a particle size in the range of 1-10 microns and more preferably 2 to 5 microns. The immobilizer is preferably selected from porous zinc oxide or calcium carbonate.

(8) The inorganic porous materials that have nano plate like structures are preferably obtained from a commercial source or can be prepared by a standard precipitation process that promote formation of porous structures, by using crystal habit modifiers or by precisely controlling the precipitation conditions. Porous calcium carbonate used is preferably vaterite polymorph and porous zinc oxide is preferably prepared by standard precipitation process that promotes the formation of porous structures, by using crystal habit modifiers. Porous magnesium hydroxide is preferably obtained from calcination of light magnesium carbonate and such materials are also commercially available.

(9) The antimicrobial particulate composition comprises 0.05% to 3% by weight of antimicrobial metal particles preferably immobilized in 97 to 99.95% by weight of an immobilizer comprising inorganic porous material selected from zinc oxide, magnesium hydroxide or calcium carbonate. Preferably the antimicrobial particulate composition comprises 0.5% to 1.5% by weight of antimicrobial metal particles and 98.5% to 99.5% by weight of an immobilizer. In general when reference is made to immobilized antimicrobial particles or immobilized antimicrobial metal particles or immobilized metal nanoparticles or immobilized materials with reference to specific metals such as silver or copper, it refers to antimicrobial particulate composition of the invention.

(10) According to the present invention there is provided process for preparing an antimicrobial particulate composition and the process comprising the steps of: i. mixing an aqueous dispersion of an immobiliser, wherein the immobiliser is present in an amount of 1-5 wt % (by weight of the dispersion) and wherein the immobilizer is selected from inorganic porous material selected from zinc oxide, magnesium hydroxide or calcium carbonate and wherein the immobiliser has a particle size in the range of 1-10 microns and an aqueous solution of a reducing agent, wherein the reducing agent is present in the aqueous solution in an amount of from 10 to 30 wt % by weight of the immobilizer in the aqueous dispersion; ii. raising the temperature of the mixture resulting from step (i) to a temperature in the range 70 C. to 90 C.; iii. adding a water soluble metal salt to the mixture resulting from step (ii) in an amount equivalent to 0.05 to 3 wt % of the metal by weight the immobilizer under mixing.

(11) The antimicrobial particulate composition prepared by the process according to the invention is preferably recovered by separating the solid from the liquid medium by preferably filtration and the antimicrobial particulate composition was preferably dried at a temperature in the range of 10-80 C.

(12) A calcination step is preferably not required for the process according to the invention and preferably the process does not comprise a calcination step.

(13) Immobilizer:

(14) The immobilizer used in the process is in the range of 1-5% and more preferably 2 to 4% and is selected from inorganic porous material selected from porous zinc oxide, magnesium hydroxide or calcium carbonate. The particle size is in the range 1-10 microns and more preferably 2 to 5 microns. The immobilizer is used as an aqueous dispersion. It is preferred that the immobilizer is selected from porous zinc oxide or calcium carbonate.

(15) Reducing Agent:

(16) The reducing agent is preferably selected from water soluble salt of a carboxylic acid with a 1-4 carboxylate group and more preferably it is selected from sodium acetate, sodium oxalate, trisodium citrate or disodium ethylene diamine tetra acetate. It is preferred to include trisodium citrate as the reducing agent. The reducing agent is used in the range 10-30% by weight with respect to immobilizer and more preferably 15 to 20% by weight with respect to immobilizer. The reducing agent is preferably provided as an aqueous solution.

(17) It is essential that the aqueous dispersion of the immobilizer and the aqueous solution of the reducing agent are mixed before adding the metal salt solution. The temperature of the mixture is raised to a temperature in the range 70 C. to 90 C. and more preferably 80 to 85 C.

(18) The pH of the reaction medium during the process is maintained preferably at pH greater than 5 and more preferably is in the range 6-8.

(19) Water Soluble Metal Salt:

(20) It is preferable that the water soluble metal salt is selected from water soluble salt of silver or of copper. It is preferable to add the water soluble metal salt as an aqueous solution.

(21) It is preferable that the water soluble silver salt is selected from silver nitrate or silver acetate. 0.05% to 3% of silver by weight of an immobilizer that is delivered preferably through an aqueous solution is preferably silver nitrate or silver acetate. To deliver the required % weight of silver, the range of silver nitrate preferably is 0.08% to 4.72% by weight or 0.078% to 4.63% by weight of silver acetate. It is preferred to use silver nitrate as the water soluble salt of silver.

(22) It is preferable that the water soluble copper salt is selected from copper (II) sulphate, copper (II) nitrate, copper (II) chloride, and copper (II) acetate. 0.05% to 3% of copper by weight of an immobilizer is delivered preferably through an aqueous solution of copper salt in the range between 0.1% to 9%, more preferably in the range of 0.13% to 7.5% for copper (II) sulphate, 0.15% to 8.9% for copper (II) nitrate, 0.1% to 6.4% for copper (II) chloride and 0.14% to 8.6% for copper (II) acetate. It is preferred to use copper (II) nitrate as the water soluble salt of copper.

(23) According to yet another aspect of the present invention there is provided a personal care or hygiene composition comprising: i. 5% to 85% by weight of a surfactant and ii. 0.1 to 5% by weight of an antimicrobial particulate composition obtainable by a process in accordance with the present invention.

(24) The level of the surfactant in the personal care composition is 5% to 85% by weight of the personal care or hygiene composition and preferably 15-40% by weight.

(25) The surfactant is selected from class of anionic, non-ionic, cationic or zwitterionic surfactants and preferably selected from anionic surfactants. The anionic surfactant is preferably selected from soap or non-soap surfactant.

(26) It is preferred that the antimicrobial particulate composition is 0.1 to 5% by weight of the personal care composition or hygiene composition and more preferably 0.25 to 4% by weight of the personal care or hygiene composition.

(27) The personal care or hygiene composition of the present invention can be in the form of liquid or solid compositions. Non-limiting examples of such topical compositions include leave-on skin lotions and creams, antiperspirants, deodorants, lipsticks, foundations, mascara, sunless tanners or sunscreen lotions, and wash-off products like shampoos, conditioners, shower gels, or toilet bars.

(28) It is preferable that the composition comprises conventional ingredients used in a personal care or hygiene composition and more preferably ingredients such as fluorescers, perfumes, texture controlling agents, emollients, and other antimicrobial agents.

(29) The invention will now be illustrated by means of the following non-limiting examples.

EXAMPLES

Example 1

(30) Preparation of antimicrobial particulate composition comprising silver nanoparticles

(31) Preparation of the Immobilizer:

(32) Micron sized particles of the immobilizer were prepared by the process described below: i. Porous zinc oxide:

(33) Porous zinc oxide was prepared by a process as described in CrystEngComm 15, 32 (2013), pp. 6349-6358. For preparing 100 g of porous zinc oxide, 60 g of zinc nitrate and 140 g of hexamine was mixed with trisodium citrate, where the ratio of the concentration of zinc to the concentration of trisodium citrate was maintained at 10, and the mixture was heated in a stoppered hydrothermal container at 90 C. for 12 hours. The precipitated porous zinc oxide was filtered, washed with deionised water and dried in air. ii. Porous Calcium carbonate:

(34) 100 g of porous calcium carbonate was prepared by mixing 147 g of calcium chloride with 106 g sodium carbonate under constant stirring at 10 C. for 5 hours. The slurry was aged for 5 hours, filtered, washed with deionised water and dried in air. iii. Porous magnesium hydroxide:

(35) Magnesium oxide of light, pure grade was purchased from Merck. Magnesium oxide was dispersed in water to get porous magnesium hydroxide which was used as the immobilizer.

(36) Preparation of the Antimicrobial Particulate Composition Comprising Silver Nanoparticles:

(37) 400 mg of immobilizer which was porous zinc oxide or calcium carbonate or magnesium hydroxide was dispersed in 10 mL water and mixed with 4 mL of 1% trisodium citrate solution. The mixture was heated to 80 C. followed by the addition of freshly-prepared 0.25 mL of 2% silver nitrate. The mixture was stirred for 20 minutes at 300 rpm. The sample was filtered and dried at room temperature. The size of the silver nanoparticles formed in the above process ranged between 5-50 nm and the silver nanoparticles get immobilized in the respective immobilizer.

(38) Evidence for Silver Incorporation in the Immobilizer:

(39) Energy-dispersive X-ray spectroscopic analysis of antimicrobial particulate composition comprising silver nanoparticles and porous zinc oxide prepared by the process described in Example 1 was carried out using a FESEM instrument (ZEISS) operated at 10 kV electron voltage to determine the presence of silver in the antimicrobial particulate composition. The analyzed weight % of different ingredients is shown in table 1 below:

(40) TABLE-US-00001 TABLE 1 Element Weight (%) Oxygen 23.60 Zinc 73.09 Silver 3.31 Total 100.00

(41) Results in table 1 show that the antimicrobial particulate composition contains zinc, oxygen and silver as constituents.

(42) Reflectance of the Antimicrobial Particulate Composition Comprising Silver Nanoparticles:

(43) Reflectance of antimicrobial particulate composition comprising silver nanoparticles where the immobilizer used was zinc oxide, calcium carbonate or magnesium hydroxide and prepared as described in Example 1, was measured.

(44) In the comparative example (Example 5) where commercial zinc oxide which was not porous was used to immobilize the silver nanoparticles, otherwise using the same process. In another comparative example (Example 6), a process according to prior art was used where the immobilization of the silver particles was done by calcination. In this process the porous zinc oxide was used without using the reducing agent and at the end of the process the material was calcined by heating it to a temperature of 500 C.

(45) Reflectance of the materials described above were measured using Gretag Macbeth reflectometer at 460 nm in the SCI, SAV mode, UV excluded condition. A film of the dispersion of immobilized silver nanoparticles was prepared on a glass slide having an area of 4 cm4 cm. The film was dried and reflectance was measured. The b* values measured indicate the dullness index. Negative b* values indicate the closeness to whitish colouration and b* values less than 0.5 indicate near whitish coloration. The data is presented in table 2.

(46) TABLE-US-00002 TABLE 2 Examples Material b* Example 2 Silver nanoparticles immobilized in porous zinc 1.1 oxide Example 3 Silver nanoparticles immobilized in porous 8.0 calcium carbonate Example 4 Silver nanoparticles immobilized in porous 1.9 magnesium hydroxide Example 5 Silver nanoparticles immobilized in non- 5.0 porous rod shape zinc oxide Example 6 Silver nanoparticles immobilized in porous zinc 3.6 oxide by a process of calcination

(47) The data presented in table 2 show that antimicrobial particulate composition prepared according to the invention have a negative b* value indicating a better desired color profile which can be used in the formulations without affecting the product aesthetics. In the comparative examples where the immobilizer used was non-porous zinc oxide and had a rod shaped structure or where the process of immobilizing silver particles was done by a calcination process, the immobilized materials had a relatively high b* values indicating a dark colouration.

(48) Variation in Process Parameters of Efficacy:

(49) Preparation of the antimicrobial particulate composition comprising silver nanoparticles was done by changing the concentration of the immobilizer, silver nitrate or trisodium citrate. For all these experiments, zinc oxide was used as the immobilizer.

(50) The reflectance of the antimicrobial particulate composition comprising silver nanoparticles obtained was measured as described above to evaluate the b* value to indicate the coloration of the material and the results are presented in table 3. In table 3, under the process condition column, the weight % of porous zinc oxide is with respect to the volume of water while the percentages of silver and trisodium citrate are with respect to the porous zinc oxide. All percentage values mentioned in table 3 correspond to amount of ingredients used during preparation of antimicrobial particulate composition comprising silver nanoparticles. The product composition column indicates the weight % of silver and porous zinc oxide after immobilization of silver nanoparticles.

(51) TABLE-US-00003 TABLE 3 Process condition Product Porous Trisodium composition Example Process zinc oxide Silver citrate Ag ZnO No. variation mg wt % mg wt % mg wt % wt % wt % b* 7 Variation 250 2.5 6.35 2.5 40 16 2.5 97.5 5.92 8 of porous 400 4 6.35 1.6 40 10 1.6 98.4 5.36 9 zinc oxide 1000 10 6.35 0.6 40 4 0.6 99.4 0.45 concentration 10 Variation 400 4 1.27 0.3 40 10 0.3 99.7 12.3 8 of silver 400 4 6.35 1.6 40 10 1.6 98.4 5.36 11 concentration 400 4 25.41 6.0 40 10 6.0 94.0 +13.9 12 400 4 63.53 13.7 40 10 13.7 86.3 +1.1 13 Variation 400 4 6.35 1.6 20 5 1.6 98.4 +3.84 8 of trisodium 400 4 6.35 1.6 40 10 1.6 98.4 5.36 14 citrate 400 4 6.35 1.6 120 30 1.6 98.4 10.9 concentration

(52) The data presented in table 3 shows that variation in process parameters during the preparation antimicrobial particulate composition using silver nanoparticles and porous zinc oxide. The b* value becomes more positive with increase in porous zinc oxide weight % (Example 9) while increase in silver weight % leads to positive b* value (Example 11 and 12). Alternatively, lowering of trisodium citrate weight % below 10 leads to positive b* value (Example 13). Overall for most optimal result as shown in table 3 for obtaining b* value less than 0.5, the aqueous weight percentage of porous zinc oxide is in the range of 1-5, and the weight percentage of citrate and silver with respect to porous zinc oxide is in the range of 10-30 and 0.05-3 respectively.

(53) Sequence of the Process Steps:

(54) The sequence of addition of materials according to the present invention where the aqueous dispersion of the immobilizer is mixed with the aqueous solution of a reducing agent such as trisodium citrate followed by the addition of the antimicrobial silver nitrate was compared with a process where the antimicrobial silver nitrate was added before the addition of the reducing agent or where the process of preparation of porous zinc oxide as described earlier in Example 1 was carried out in the presence of silver nitrate otherwise using the same process. In the processes the levels of the various materials were as in Example 8.

(55) The reflectance of the immobilized material obtained was measured as described above to evaluate the b* value to indicate the coloration of the material and the results are presented in table 4.

(56) TABLE-US-00004 TABLE 4 Example Process b* Example 8 Addition of trisodium 5.4 citrate to porous zinc oxide followed by silver nitrate Example 15 Addition of silver nitrate +13.5 to porous zinc oxide followed by trisodium citrate Example 16 Hydrothermal treatment +12.6 of zinc nitrate and hexamine in the presence of trisodium citrate and silver nitrate

(57) The data presented in table 4 show that the addition of trisodium citrate to zinc oxide followed by silver nitrate yields antimicrobial particulate composition according to the invention with negative b* value (Example 8) while the addition of silver nitrate to zinc oxide followed by trisodium citrate yields positive b* value (Example 15). Also, formation of porous zinc oxide in the presence of silver nitrate yields again positive b* value (Example 16).

(58) Antimicrobial Efficacy:

(59) The antimicrobial efficacy was tested using gram positive bacteria Staphylococcus aureus and gram negative bacteria Escherichia coli bacteria by using Time kill assay according to BS EN1040 protocol.

(60) 10 g soap was dissolved in 90 mL water at 50 C. It was allowed to equilibrate to a temperature of 45 C. in a water bath. Antimicrobial particulate composition comprising silver nanoparticles was added to the soap solution to get 1 ppm effective silver load and the same was calculated by ICP-OES analysis. 1 mL bacterial suspension of Staphylococcus aureus (10.sup.8 cells/mL) and E. coli (10.sup.8 cells/mL), respectively, was added to the tubes containing 1 mL sterile water and was allowed to equilibrate at 45 C. for 2 minutes. 8 mL of 10% soap solution containing antimicrobial particulate composition comprising silver nanoparticles was added to these bacterial suspensions respectively and was allowed to act for 30 seconds and 60 seconds respectively. On these respective time points, 1 mL suspension was taken and added to 9 mL of neutralizer (Dey-Engley Neutralizing Broth) to inactivate the action of the antimicrobial. The neutralized samples were then plated after serial dilution in Trypticase Soy Agar (nutrient medium) to enumerate the residual bacteria. The efficacy of the antimicrobial particulate composition comprising silver nanoparticles was also evaluated with Methicillin-resistant Staphylococcus aureus (MRSA) at 60 and 300 seconds respectively. The results are presented in Table 5.

(61) TABLE-US-00005 TABLE 5 Bacterial reduction (log CFU/mL) E. coli S. aureus MRSA Example Material 30 s 60 s 30 s 60 s 60 s 300 s Example 17 Soap 3.4 3.5 0.0 0.1 0.3 1.2 Example 18 Soap + Porous 3.9 6.3 0.0 0.2 zinc oxide Example 19 Soap + Silver 6.3 7.7 4.2 5.2 nitrate Example 20 Soap + silver 3.7 7.0 4.8 6.0 nanoparticle Example 21 Soap + silver 7.6 6.7 5.1 6.7 2.0 4.6 nanoparticles immobilized in zinc oxide Example 22 Soap + silver 8.1 8.1 4.8 6.8 1.8 4.9 nanoparticles immobilized in calcium carbonate Example 23 Soap + silver 5.2 7.8 3.9 5.4 nanoparticles immobilized in magnesium hydroxide

(62) The data in table 5 shows that immobilized silver nanoparticles (Example 21-23) have superior bacterial reduction against both E.coli and S.aureus when compared to any of its individual constituents. Even though the silver nitrate and silver nanoparticle with soap shows comparative antimicrobial action, its use is limited as silver nitrate being unstable can't be formulated in products while the use of only nanoparticles has issues related to agglomeration, instability upon storage and environmental hazards. The data also shows that the immobilized silver nanoparticles have antimicrobial action against MRSA with 5 log reduction at 300 s.

(63) No efficacy validation tests were performed on the immobilizers per se, used in Example 22 and Example 23, as there are no literature evidence on antimicrobial activity of either calcium carbonate (immobilizer in Example 22) or magnesium hydroxide (immobilizer in Example 23).