F0RM2
THE PATENTS ACT, 1970
(39 of 1970)
COMPLETE SPECIFICATION (See section 10 and rule 13)
TITLE OF THE INVENTION:
"ENVIRONMENT-FRIENDLY SILVER NANOCOMPOSITE AND METHOD FOR
SAME"
2. APPLICANTS
i. NAME : HERLEKAR; PRAVIN SHIVDAS
NATIONALITY: INDIAN
ADDRESS : B-34, M.I.D.C. BADLAPUR (E), DIST-THANE,
MAHARASHTRA, PIN CODE: 421503, INDIA.
ii. NAME : MORO; RAVI VITTHAL
NATIONALITY: INDIAN
ADDRESS : B-34, M.I.D.C. BADLAPUR (E), DIST-THANE,
MAHARASHTRA, PIN CODE: 421503, INDIA.
The following specification particularly describes the invention and manner in which it is to be performed

FIELD OF THE INVENTION
The present invention relates to environment-friendlysilver nanocomposite and method to prepare the silver nanocomposite and its application.The invention further relates to conjugation of a silver nanoparticle with eco -friendly thickening and reducing agent.
BACKGROUND AND PRIOR ART:
Preparation of silver nanoparticles has been physical and chemical methods, they are there with high production cost and environmental pollution, while some residual in the product are harmful for prolong effect as they are chemical ingredients.
The preferred method of preparing nanoparticles is chemical method wherein, mostly strong reducing agent such as NaBH4, hydrazine, hydroxylamine are used. Common dispersants or stabilizers polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), sodium lauryl sulfate (DBS) and a different chain length of thiol molecules or molecules containing a mercapto group, etc. are in use, but these agents typically related to environmental toxicity and bio-toxic.
At present, the "green chemistry" has gradually become an important theme in the field of chemistry. As the need for environmental protection, chemical processes are considered with focused and committed to minimize chemical contamination. The use ofenvironment friendly, non-toxic chemicals are being used to prepare the silver nanoparticles, therefore, a method to prepare nano particle utilizes selecting natural raw materials, with simple and easy step with reduced pollution.

Various methods have been reported over the last two decades for the synthesis of silver nanoparticles which involve the reduction of metal salts with a chemical reducing agent, such as sodium citrate, sodium borohydride, or other organic compounds. As referred in Plyuto, Y.; Berquier, J.- M; Jacquiod, C; Ricolleau, C, Ag Nanoparticles synthesized in template-structured mesoporous silica films on a glass substrate. Chemical Communications {Cambridge) 1999, 1653-1654. Also, Rivas, L.; Sanchez-Cortes, S.; Garcia-Ramos, J. V.; Morcillo, G., Growth of silver colloidal particles obtained by citrate reduction to increase the Raman enhancement factor. Langmuir 2001, 17, 574-577. See Tan, Y.; Jiang, L.; Li, Y.; Zhu, D., One Dimensional Aggregates of Silver Nanoparticles Induced by the Stabilizer 2-Mercaptobenzimidazole../. Phys. Chem. S2002, 106, 3131-3138. See Zhang, Z.; Patel, R. C,; Kothari, R.; Johnson, C. P.; Friberg, S. E.; Aikens, P. A., Stable silver clusters and Nanoparticles prepared in polyacrylate and inverse micellar solutions..J. Phys. Chem. B 2000, 104, 1176-1182.
The mechanism for the antimicrobial property of silver is only partially understood. It has been hypothesized that the positively charged Ag is able to interact with the negatively charged bacteria cell wall, inhibiting membrane permeability (see Ratte, H. T., Bioaccumulation and toxicity of silver compounds: a review, Environ. Toxicol. Chem. 1999, 18, 89-108), inactivating necessary enzymes by interaction with the thiol groups of the proteins (see Gupta, A.; Maynes, M.; Silver, S., Effects of halides on plasmid-mediated silver resistance in Escherichia coli, Appl. Environ. Microbiol. 1998, 64, 5042-5045; see also Matsumura, Y.; Yoshikata, K.; Kunisaki, S.-i.; Tsuchido, T., Mode of bactericidal action of silver zeolite and its comparison with that of silver nitrate, Appl. Environ. Microbiol. 2003, 69, 4278-4281), leading to cell death.

The smaller the particle the more surface area is exposed to water forming more silver ions which can deactivate the proteins necessary for bacteria, viruses, and fungi to survive. The slower release of silver cations from silver nanoparticles can avoid the constant delivery of an excess amount of silver to the area compared with other Ag+ based chemicals. Using silver nanoparticles, the metallic silver is not as susceptible to deactivation by the chloride molecules compared with the Ag+ (Dunn, K.; Edwards-Jones, V. The role of Acticoat with nanocrystalline silver in the management of burns; Wythenshawe Hospital Burns Unit, Manchester, UK: England: United Kingdom, 2004; pp S1 -9).
Unfortunately, using such reducing agents introduce chemicals that are biologically incompatible or environmentally toxic. Thus the conventional synthesis of silver nanoparticles incorporates contaminants that could pose problems in biomedical applications.
As evident from prior art, it becomes challenging to design and manufacture silver nano composite that is biologically compatible and environmentally non-toxic. The present invention is an improvement over the silver nano composite and its process by utilizing natural ingredient as reducing agent, colloidal agent and excipient.
The discussion of documents, acts, materials, devices, articles and the like is included in this specification solely for the purpose of providing a context for the present invention. It is not suggested or represented that any or all of these matters formed part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed before the priority date of each claim of this application.

OBJECTIVES OF THE INVENTION
• The primary objective of the present invention is to provide a silver nanocomposite which is environment- friendly.
• A further object of the invention is to provide a silver nano composite which is easy to prepare and uses naturally occurring elements.
• Another object of the invention is to provide an economic and simple process to prepare silver nanocomposite.
• The further object of the invention is to provide a silver nano composite which has wide bio-medical application because of natural content.
SUMMARY OF INVENTION
The present invention relates to an environment-friendly silver nanocomposite, and method for forming the silver nanocomposite, it further relates to an application conjugation of a silver nanocomposite. The ingredients used as reducing agent, stabilizing agent and morphological control agent to synthetize silver nano are naturally occurring agent. The environmentally-friendly method for the metal nanoparticle, which is disclosed by the invention, has the moderate reaction condition, short synthesis time and low production cost, the particle diameter of the obtained metal nanoparticle is 1-100nm.The silver nanocomposite has varied application from personal hygiene product to the product used in hospitality industry, healthcare industry an cosmetic.

STATEMENT OF INVENTION
The present in relates to an antimicrobial composite of Nano form silver ions, wherein said composite comprises nanofabricated silver ions suspended in stabilizing agent. The stabilizing agent is aloe Vera concentrate.
A process for preparation of an antimicrobial composite of Nano form silver ions suspended in stabilizing agent comprising steps;
a. Mixing silver salt in stabilizing agent
b. Addition of reducing agent at ambient temperature
c. Addition of ammonia solution under constant stirring at temperature range from 30 to 3 5°C.
The reducing agent is glucose and molar ratio of reducing agent to silver salt is 10:1. The stabilizing agent is Aloe vera concentrate in the range of l0g/ltr to 50g/ltr.A bandage, mask, cosmetic and healthcare product comprising a non-adhesive adsorbent pad with the silver nanocomposite embedded therein.
DESCRIPTION OF INVENTION
While the making and using of various embodiments of the present invention are discussed in detail below, it should be appreciated that the present invention provides many applicable inventive concepts that can be embodied in a wide variety of specific contexts. The specific embodiments discussed herein are merely illustrative of specific ways to make and use the invention and do not delimit the scope of the invention.

To facilitate the understanding of this invention, a number of terms are defined below. Terms defined herein have meanings as commonly understood by a person of ordinary skill in the areas relevant to the present invention. Terms such as "a", "an" and "the" are not intended to refer to only a singular entity, but include the general class of which a specific example may be used for illustration. The terminology herein is used to describe specific embodiments of the invention, but their usage does not delimit the invention, except as outlined in the claims.
The present invention provides a method for manufacturing silver nano-particles, which can be indifferently have large application, ranging from coating on substrate or incorporation in it while manufacturing. The substrate for application of silver nano-particles can be non-limiting to natural/synthetic fibers, polymers, glassto naturally grown ingredients like fruits, and other eatables.
The method according to the present invention comprises the reaction between silver ions and a reducing agent in an aqueous or hydro-alcoholic solution in which the substrate to be functionalized are present.
The reaction between silver ions, Ag+, and the reducing agent is carried out in the solution in which the substrate is present. The Ag+ ions can be obtained from the dissociation of a silver salt soluble in water, e.g. perchlorate, AgC104, or preferably nitrate, AgNO3. The reducing agent can be, for instance, ascorbic acid, a reducing sugar such as glucose or fructose, or a citrate, preferably aleo vera.
The solution wherein the reaction is carried out can be obtained in different ways. For instance, it is possible to introduce the desired water amount in a suitable container, dissolving the first

reactant (typically the silver salt) in water, and successively adding the reducing agent in the thus obtained solution; alternatively, it is possible to prepare two separate aqueous solutions, one with the silver salt and the other with the reducing agent, and successively combining the two solutions, preferably slowly and under stirring.
The concentration of the solutions may be different in the two cases, i.e. depending on whether one single solution, containing a first reactant which is then added with a second reactant in solid form, or two separate solutions are used.
In the first case, it is preferable that the reactant already in solution is the silver salt, and that the reducing agent is added to this solution. When this procedure is followed, the starting solution containing a silver salt has a preferred concentration from 10"4to 10"' M; concentrations lower than 10" M may reduce the coating of the substrate with silver nano-particles, while concentrations higher than 10" M may render the solution cloudy, when the reducing agent is added, and produce silver agglomerate deposits on the substrate instead of nano-particles.
Use of aloe vera and glucose project a healing and a protecting technology which is exquisitely eco-friendly and GREEN. After enhancing several health products
In one of the embodiment use of silver ions as an anti-bacterial/ anti/viral has found to be very effective specifically againstEscherichia coli,Mycobacterium tuberculosis, Streptococcus, Pseudomonas aeruginosa, Salmonella typhus, Cornebacterium Diptheria.The silver nano particles along with Aloe Vera are actively effective in prevention of microorganism growth by more than 99%.

In other embodiment the application of nano silver composite is not limited to Anti-Viral and Anti-Bacterial Masks, Water Purification Systems, Deodorizer, Sterile Sanitizers, Surgical Dressings, Nano-Gels based on Gold and Silver but also silver nanoparticles are used as antibacterial/antifungal agents in a diverse range of applications including air sanitizer sprays, socks, pillows, slippers, face masks, wet wipes, detergent, soap, shampoo, toothpaste, air filters, coatings of refrigerators, vacuum cleaners, washing machines, food storage containers, cellular phones, and even in liquid condoms. All type of mask, aprons, bed spreads, pillow covers, bandage cloth, under garments, vests, diapers, sanitary napkins, A.C filter cartridge, uniforms, curtains, carpets, gel for burns, hand sanitizers, antimicrobial creams, antimicrobial soaps, moisturizers, anti-ageing creams, facial creams, bleach creams, night energizing creams, eye cream, eye shadow, pet bottles, polymeric toys, toilet seat covers, water purification candles.
The present invention is further described with the help of the following examples, which are given by way of illustration and therefore should not be construed to limit the scope of the invention in any manner.
EXAMPLES:
Example 1:
Working solutions of Silver Nitrate and or salts thereof with concentration of range from 10"4 to 10" M were prepared from stock solution of 10" M in distilled water and Aloevera concentrate.Correct volume of working silver solution equal to the desired concentration was taken into glass flask and magnetically stirred at ambient temperature,to which was added a Glucose solution 10 times higher in concentration to Silver content.The aloevera concentration can be varied in the range of lOg/ltr to 50g/ltr to meet the final required concentration.The final

solution was mixed and stirred for about 10 minutes,then ammonia was added and stirred continuously until all Ag ions were reduced into the nano dimensional state.The process temperature was maintained between 30-35°C during reduction.
The resultant silver colloid dispersion was analyzed and the morphology characterized by transmission electron microscopy(TEM,JOEL 2000EX,Japan) operated at 200 kv,scanning electron microscopy (SEM,Model S-4700I,H.itachi,Japan) and X-ray diffraction (XRD,MXP 18,MAC Science,Japan).
Also, particle size and distribution were determined by counting over 200 particles from TEM image using Software Image J (NIH,USA).The (Ag+) ion concentration was traced by Cole-Parmer silver electrode(USA) during the reaction.
The results obtained thus comprised the average size of 22nm with a standard deviation of 4.7nm after separation and washing procedures.
The conversion under these conditions was nearly complete at about 98%.
The so obtained Silver colloid dispersion was diluted to nearly 11 times using distilled water to obtain a final concentration of 50ppm.
This solution was then sprayed uniformly onto a viscose fabric of 125 denier and left to dry. The dried fabric was then cut and stitched to the required specification resulting in an overall concentration of 10 ppm per sq.mtrof the fabric.
Examples: General Antibacterial Assay
The antibacterial activities of different woven materials according to the invention were tested towards effectiveness to inhibit the growth of Gram-negative Escherichia coli as well as against the Gram-positive Staphylococcus aureus (S. aureus, specifically methicilin-resistant

staphylococcus aureus (MRSA) bacteria), applying the ASTM:E 2149-10 and ASTM E2149-13a method.
The ASTM:E2149-10 and ASTM E2149-13a method is a quantitative test method to determine the antibacterial activity of antibacterial finished textile products.
Example 1 : Antimicrobial activity
The textiles treated with nano silver solution were subjected to anti-bacterial tests according to ISO 20743:2007 (Textiles - Determination of antibacterial activity of antibacterial finished products).
Test Organism Used: Staphylococcus aureus ATCC 6538 (2.10 X 105 CFU/ml)
Quantity of Sample: 1 gm
Method: Fabric pieces in contact with bacterial suspension on wrist arm shaker for 24 Hr
Evaluation of antimicrobial activity of fabric sample by ASTM E2149-13a
Type of Sample Description of Inoculated Inoculated Percentage
sample sample at sample at 24 Reduction of
0 hours (B) hours (A) Bacteria
Socks Light Gray colour 1.89 X 10s < 10 >99.99
Bath Towel Biscuit colour 1.92X10'' <~10 >99.99
Bed sheet White colour 1.93 XI05 1.00 X 102 >99.94
T-Shirt Yellow colour 1.93 XI05 99.99
Undergarment Light Pista colour 1.92 XI03 50 >99.97
Mask Light Blue 1.94 X 10s '3.30X 10" >99.82
Table .1 : Antibacterial Activity by evaluation of number of viable cells [Log CFU] for Staphylococcus aureas.

Where,
CFU: Colony Forming Unit= No. of Organisms
Percentage reduction of microorganism: ( R)= 100(B-A/B)
Example 2:
Test Organism Used: Escherichia Coli ATCC 11229 (1.90 X 105 CFU/ml)
Quantity of Sample: 1 gm
Method: Fabric pieces in contact with bacterial suspension on wrist arm shaker for 24 Iir
Evaluation of antimicrobial activity of fabric sample by ASTM E2149-1.0

Type of Sample Description of sample Inoculated sample at 0 hours (B) Inoculated sample at 24 hours(A) Percentage Reduction of Bacteria
Socks Light Gray colour 1.75 X 10' 1.81 X 104 89.65
Bath Towel Biscuit colour 1.92 X 10* 5.10 X102 99.73
Table 2 : Antibacterial Activity by evaluation of number of viable cells [Log CFU] for Escherichia Coli

Example 3:
Test Organism Used: Staphylococcus aureas ATCC 6538 (2.20 X 105 CFU/ml) Quantity of Sample: 1 gm
Method: Fabric pieces in contact with bacterial suspension on wrist arm shaker for 24 Hr Evaluation of antimicrobial activity of fabric sample by ASTM E2149-10

Type of Sample Description of sample Inoculated sample at 0 hours (B) Inoculated sample at 24 hours (A) Percentage Reduction of Bacteria
Bed sheet White 2.10 X105 <10 99.99
Curtain Green 1.97 X 105 20 99.98
Spread Sheet Green 1.97 X 105 20 99.98
Cup Mask Light Blue 1.94X 105 3.30 X102 99.82
Undergarment Black 1.83 X 105 <10 99.99
Table 3: Antibacterial Activity by evaluation of number of viable cells [Log CFUj for Staphylococcus aureas.
Example 4: Testing of Antimicrobial activity after washing
The solution as disclosed in present invention was sprayed on White polyster fabric and dried. The obtained materials subjected to anti-bacterial tests according to ISO 20743:2007 (Textiles -Determination of antibacterial activity of antibacterial finished products). This test is used to quantitatively measure the antimicrobial activity of antibacterial finished textile products.

Strains tested were: Klebsiella pneumoniae AATCC 147:2004 Gram Negative bacteria and Staphylococcus aureus AATCC 147:2004 Gram Positive bacteria

SrNo. Bacteria Zone of Inhibition for Initial Sample Zone of Inhibition for sample After 20 Wash cycle
1 AATCC 147:2004 Gram Negative bacteria 16.1 mm 15.2 mm
2 AATCC 147:2004 Gram Positive bacteria 14.9 mm 14.2 mm
Table 4 : Antibacterial Activity after multiple laundry cycles
All of the compositions and/or methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the compositions and/or methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

CLAIMS
We claim,
1. An antimicrobial composite of nano form silver ions, wherein said composite comprises nanofabricated silver ions suspended in stabilizing agent.
2. An antimicrobial composite of nano form silver ions as claimed in claim 1, wherein stabilizing agent is aloe Vera concentrate.
3. A process for preparation of an antimicrobial composite of nano form silver ions suspended in stabilizing agent comprising steps;
d. Mixing silver salt in stabilizing agent
e. Addition of reducing agent at ambient temperature
f. Addition of ammonia solution under constant stirring at temperature range from30 to 35°C.
4. A process for preparation of an antimicrobial composite as claimed in claim 3, wherein reducing agent is selected from glucose.
5. A process for preparation of an antimicrobial composite as claimed in claim 3, wherein molar ratio of reducing agent to silver salt is 10:1.
6. A process for preparation of an antimicrobial composite as claimed in claim 3, wherein stabilizing agent is selected from Aloe vera concentrate.
7. A process for preparation of an antimicrobial composite as claimed in claim 3 and 6, wherein aloe vera concentrate is l0g/ltr to 50g/ltr .

8. A bandage, mask, cosmetic and healthcare product comprising a non-adhesive adsorbent pad with the silver nano composite of claim 1 embedded therein.