LV13745B - Silver/water, silver gels and silver based compositions, and methods for making and using the same - Google Patents

Abstract

This document discloses a colorless composition comprising metal particles (e.g., silver nanoparticles) and water. Said particles comprise an interior of elemental metal (e.g., silver) and an exterior of metal oxide (e.g., one or more silver oxide(s)). And the metal nanoparticles are present in the water at a level of about 5-40 ppm, and wherein the composition manifests significant antimicrobial properties. Methods of use of the composition are described. Such compositions can be incorporated into a hydro-gel with essentially no loss of antimicrobial properties. In this document are also disclosed various metal-containing compositions with unexpected biological efficacy are also disclosed.

Description

Silver / water, silver gels and silver-based compositions and methods for their preparation and use (1) Scientific field.

(2) The current product is predominantly based on an unprecedented silver / water composition (sometimes referred to as silver nanoparticles dissolved in water), more specifically, based on unprecedented compositions and / or silver / water structures, silver hydrogels, novel silver compositions with advanced antibiotics and various ligands associated with silver ions, silver gels based on certain parent silver / water compositions, as well as silver ions and / or metal (s) bound / present in certain clathrates such as clays and / or zeolite materials, and is based on methods for preparing and using such compositions as agents against various organisms (including certain viruses) that are hazardous to health or endanger humans and / or animals, as well as other organisms. In addition to silver, this document also describes other materials that can be used with silver. The various compounds and concentrations of the compositions formed are also included.

(3) Description of the prototype.

(4) Certain silver preparations are known to have bactericidal properties. Before the development of modern antibiotics, silver was used as a bactericide and antibiotic. Some centuries ago, in order for the body to absorb silver, people poured silver particles or immersed whole pieces of silver in drinking water. It is possible that the practice of consuming silver tools (eg silverware) was based on the assumption that silver had the substance needed by the body.

(5) There are several reasons why silver in solution improves a person's health. Such a solution is likely to inhibit the development of bacteria, viruses and other undesirable organisms. It is also possible that silver compositions have anti-inflammatory properties that can reduce swelling, burn complications and certain asthma symptoms.

(6) The first version of the current product describes the use of a silver composition in water to process human (or, in particular, animal) nutrition. One embodiment of the article contains a silver composition consisting of silver nanoparticles (e.g., most of these particles have a diameter in the range of 10-50 nanometers) and, optionally, may comprise silver metal parts and an outer coating or non-specific part (e.g. ionic silver coating, coating (s) of one or more silver oxides (e.g., different compositions and / or different phases, etc.) suspended in water (e.g., purified water). In a further preferred embodiment, at least 90% of these particles are 10-50 nanometers in diameter The preferred product formulation contains a silver composition consisting of silver particles (including certain silver oxide coated particles), more than 50% of the particles being less than 0.015 micrometres, and they are colloidally suspended (e.g. immersed) in water Another preferred product variant contains the same parts, about 95% of the particles ether is from 10-40 nanometers In a further preferred embodiment, about 95% of the particles have a diameter of 10-30 nanometers.

(7) Product summary.

(8) The existing product is predominantly aimed at using silver at a water level of 5 to 10 ppm (in some cases less than 5 ppm) to kill or render harmless microorganisms (including various viruses) dangerous to humans and / or animals, and other living organisms. The existing product is specifically devoted to compositions containing silver nanoparticles. In the preferred embodiment of said parts, such as the original silver particles and the outer coating or partial coating, as well as a layer for one or more metal oxides (e.g., ionic silver oxide, silver oxides such as: Ag ₂ O, AgO, Ag ₄ O ₄ , etc. .). Said coatings are of various states (e.g., Ag ₂ O is moniclinic and / or tetragonal) and are present in water in which the silver particles are suspended in a suspension (e.g., colloidal suspension) at a level of 5-40 ppm. One variation of the present article contains silver nanoparticles (hereinafter referred to as "silver particles" or similar, when treated in accordance with the electrochemical techniques described herein, and not limited to the original silver particles, which may be partially or substantially single or multiple composition coatings, such coatings containing one or more silver oxides, or at least parts thereof, in water (preferably purified water, to be described later) at a concentration of 5 to 40 ppm, more than 50% silver particles having a particle size of less than 0.015 micrometers. In the preferred embodiment, most particles have a diameter of 10-30 nm. Silver composition in water (as well as silver particles obtained as completely different parts from the silver / water composition made according to the product) as well as silver / water compositions made according to the product ideas for subsequent preparation of gel, powder, clay or zeolite (as described later in the document), it is formulated according to the product, for example, effective antibacterial agents and antiviral agents (in some cases also antiparasitic agents). This product is directed to silver compositions containing 5-40pmm silver in water, and to the appropriate silver / water methods set forth herein, these compositions are effective as antibacterial agents when used as follows: (1) orally in living organisms; (2) externally in living organisms as well as externally (or internally) on various surfaces both solid and porous (eg kitchen table, cooking surfaces, cooking utensils, various surfaces in hospital, medical supplies, water lines in both metal and plastic) , air filtration units, etc.); (3) mixing silver or silver-containing water compositions with contaminated water (eg, effluent treatment, pond water, contaminated water tanks, water lines, etc., preferably sampled prior to mixing to yield a water purification procedure).

(9) One preferred embodiment of the product is directed to silver compositions in water made using a modification of the apparatus and / or methods described in U.S. Pat. No. 6,214,2999 (Patent '299'), incorporated herein by reference. Additionally, metal compositions such as, for example, copper (and copper alloys), zinc, platinum, titanium alloys, and compounds may be used on washable metals / mixtures in accordance with product-effective methods.

(10) The process of the Machine and Patent '299 has been redesigned and improved to provide a silver blend of an existing product, which is described in more detail herein. The eighth silver / general electrode assembly as described in Patent '299 has been modified and adapted to accommodate a larger (e.g., 75-85 gallon) water tank. To start producing a silver / water blend in a 75-85 gallon tank, you must place 7075 gallons of relatively high purity water (eg filtered water, reverse osmosis water, or relatively low contamination, etc.) in the tank containing less than 2ppm of all dissolved solids, water containing less than a ppm of all dissolved solids is preferred. Preferably, about 5 gallons of the silver / water mixture, made before the commencement of major production, is added to this mixture. This 5 gallon ready mix ”is useful but not mandatory. Preparation involves a significant amount of the manufacturing material contained in the containers so that the current can flow between the different electrodes so that the required voltage / current is reached in a relatively short period of time. This Preparation "produces relatively small initial" Taylor cones ", which are described below. The water tank space is equipped with an air stream (usually located near the end of the water tank), which allows the air bubble stream to flow through the silver / water fluid during production. This approach has been found to visibly improve mixing when compared to paddle wheel mixing as described in Patent '299.

(11) This electrode assembly is powered (at least initially) with approximately 10,000 volts AC (each electrode assembly has a separate power supply) as described in Patent '299. Voltages in excess of 10,000 volts can produce solids containing a significant amount of dissolved silver ions. This mixture contains more than 97% of a metallic silver moiety with 5-40ppm and relatively low silver ions in the silver / water solids.

(12) The silver concentration is determined according to the methods described below. The 75 gallon silver / water treatment unit is run continuously and samples are analyzed from the device until the desired silver ppm concentration is achieved. It has been determined that, under the operating conditions described in the document, one and a half days of operation are required to obtain a 10 ppm silver / water composition. It takes approximately three days for the equipment to produce 22 ppm silver / water composition. It takes about six days to get the 32 ppm silver / water composition. The formation of silver particles in the silver / water composition becomes slower when a higher concentration of silver particles is used. It takes a relatively long time to increase the silver concentration in the silver / water composition to above 50 ppm. According to the processing parameters described in the document, the highest concentration achieved within a reasonable time is 50 pmm. It is possible to increase the concentration of silver particles, however, the effectiveness of lower concentrations of silver particles against various pathogens is so convincing that no higher concentration is required.

(13) Silver nanoparticles have the same overall size and shape in silver / water compositions and are described in more detail later in this document. Atypical for most standard "colloidal silver" compositions, its silver / water compositions are completely colorless and stable, taking into account moderate light and temperature changes without the need for additional means to provide stability (most colloidal silver prototypes require this). The parts and marketing process used to make the silver / water composition are said to differ in efficiency from other products known as "colloidal silver". The new silver / water composition, described in more detail below, exhibits a significant difference in physical properties (e.g., particle size, composition, spectroscopy, etc.) (14) The silver / water composition does not react with many materials added alone or in combination. , (1) hydrogen peroxide, (2) disodium EDTA (dinarea ethylene diamine tetra acetic acid), which can act as a silver / water booster (e.g., can make the silver / water composition more effective), (3) iodine (e.g. readily reacts with other substances in some cases) may enhance the effect of silver / water compositions on various pathogens, (4) various commercially available antibiotics (which may produce direct synergistic effects between silver / water compositions and antibiotics, creating new and desirable compositions) . With the new silver / water composition, many additional materials and compounds (eg added or supplemented) can be used to create synergies to achieve the desired effect that each material can achieve individually. In many cases (antibiotic compounds), the combined effects are synergistic and outweigh the effects of the individual materials or substances (eg 2 + 2 = 6). Of course, some of the possible impurities will be suitable for the new compound only as a top or surface treatment due to their potential internal toxicity to biological organisms (eg animals or humans). The amount of admixture required may vary depending on the circumstances, including certain deficiencies (eg viruses, bacteria, parasites, etc.) or the presence of infections and other materials in admixtures. However, the exact amount of admixture required will be included in the daily experiments of ordinary quality in this field. The concentration of the silver / water composition may affect the amount of additives required in daily experiments of ordinary quality in the art.

(15) One example of a desirable additive is hydrogen peroxide. Hydrogen peroxide is a disinfectant. Hydrogen peroxide has been found to have a synergistic interaction with the silver / water composition. Different concentrations of hydrogen peroxide are available, for example, at 30% by weight (% by volume or% by weight) or even higher concentrations. Although higher concentrations of hydrogen peroxide are usable, the preferred concentration for use in the silver / water composition is 30% or less and corresponds to a grade of about 1 to 5% by weight.

(16) One preferred embodiment of the present study is directed to compositions containing 5-40ppm silver particles and 1-3% by weight of hydrogen peroxide, the remainder of the composition being water (e.g., filtered or purified water). Another preferred method is to use the compositions as antimicrobial agents containing 10-40ppm silver and 1-3% hydrogen peroxide in water.

(17) Another example of an additive that combines favorably with a silver / water composition is the dinarric ethylene diamine tetra acetic acid known as "Sodium".

EDTA 'or' Disodium EDTA '(referred to in the specialized literature) having the following chemical formula: (CH ₂ N (CH ₂ COOH) CH ₂ COONa) 22 H ₂ O. Alternatively, a small amount of disodium EDTA (0.5-5pmm or more preferably 0.5ppm) is added to the silver / water compositions. In this case, a small amount of disodium EDTA increases the properties of the silver / water composition (eg, increases bactericidal, disinfectant and / or antibacterial properties). Without being bound by any particular theory or explanation, it is possible that disodium EDTA increases the permeability of cell walls, which may increase the overall effectiveness of the silver / water composition. Another variant of this study relates to a method of using the compositions in water as an antibacterial agent, antiviral agent and / or disinfectant containing 10-40ppm silver and 0.5-1 Oppm disodium EDTA.

(18) Another example of an additive that positively combines with the silver / water composition is iodine. Iodine is used in medicine as a preventive agent for the treatment of various pathogens. Various concentrations of iodine are available, with 10% iodine predominantly used. In this variant of the study, the synergistic composition contains about 20-25% by volume of the silver / water composition as a substitute for 10% solid iodine substitution. Various reactions are possible between the silver / water compound and iodine. The results of the experiments described below indicate that the silver / water synergistic composition with iodine may act as a disinfectant (eg ointment) and / or as a preventive agent to prevent infection in the case of cut wounds, burns and scratches. Another variation of this study relates to a method of using the compositions in water as an antibacterial agent, an antiviral agent and / or a disinfectant containing 10-40ppm silver and iodine.

(19) Another preferred study option contains silver / water compositions combined with various commercially available antibiotics known as coupling therapy. Coupling therapy has aroused great interest as antibiotic resistance has become widespread in the last two decades, thus causing concern worldwide. Infections caused by Gram-negative bacteria such as Escherichia coli, Klebsiella, Proteus, Shigella, and Pseudomonas have become a cause for concern because of the resistance of various drugs to antibiotics. A recent study to investigate the degree of resistance of gram-negative, clinically purified, infectious agents shows that most of the purified substances are resistant to antibiotics such as ampicillin, gentamycin, chloramphenicol, co-trimoxazole and first- and second-degree cephalosporin. . About 70% of these substances are resistant to ciprofloxacin. At this stage of the study, silver / water compounds (bonded as a liquid or dried and added as solids to form a powder, sometimes called "silver dust") combined with various antibiotics showed synergistic properties rather than additives. Quality tests showed certain antibiotics combined with silver / water formulations produced several times more effective antibiotics than individual silver (e.g., silver / water formulations combined with amikacin and cefoperazone showed an FIC of approximately 0.1875 compared to two antibiotics , which are linked to each other to give a FIC of 0.625. Both compounds were used against, for example, MRSA (Methicillin-resistant Staphylococcus aureus)), which will be described in more detail below. Another embodiment of this product relates to a method of using the compositions in water as an antibacterial agent, an antiviral agent and / or a disinfectant containing 10-40ppm silver and various antibiotics known as "compound therapy". The exact amount (and concentration) of silver / water composition that may be added to standard antibiotic therapy according to this study is a routine experiment. Specifically, a specific course of antibiotics (including antibiotic efficacy against a pathogen) that affects the amount and concentration of silver / water composition required is used to treat specific disease.

(20) Although many tests using both silver / water solids and their various additives have been described below, it has been shown that certain binders can significantly improve the results obtained with silver / water solids in various states. It has been found that forming a water-containing silver / water composition as a medium-hard hydrogel (hereinafter called "Silgel" or, in other embodiments, "Silderm") or even strips of such materials significantly improves efficiency. Hydrogels are water-absorbing gels created by adding certain water-absorbing organic polymers to an aqueous liquid. In this case, the solution should contain a patentable silver / water solution. However, it is anticipated that other "colloidal silver" solutions may be formed as hydrogels as described herein. While such hydrogels may not be as effective as those made with an existing product, they can be used properly. This product includes certain aspects of such hydrogels. Hydrogel improves the durability of silver on surfaces such as injured skin. When treating a wound, hydrogel or sheet material has the significant advantage of protecting tissue around the wound and preventing it from drying out. These factors are often important for wound healing. The hydrogel does not significantly inhibit the antibacterial properties of silver nanoparticles. Hydrogels are great hand or skin cleansers and also protect the skin (eg when hands have a hydrogel applied and the hands come in contact with pathogens, the gel helps prevent infections such as cuts or abrasions so it acts as a preventative) useful in healthcare.

(21) Clean hands are a key factor in preventing the spread of dangerous microbial and antibiotic resistance in healthcare settings. Hygienic hand cleaners used in modern medicine are alcohol based and have some disadvantages. One of these disadvantages is the damage to the skin caused by the independent use of alcohol-based disinfectants. In some cases, (1) irritating redness of the skin, and (2) known cases of redness of the skin caused by an allergy. These factors reduce the use of hand hygiene products in healthcare.

(22) Another reason for the lack of widespread use of hand detergents is that they are only mounted above sinks and wash basins. Thus, medical staff must go to the area where the sink is located to wash their hands and then go to the next patient. If hand disinfection were possible by applying the gel to the hands, this would reduce the problem. The hydrogel products of this product reduce the number of bacteria in the body over a long period of time and constitute an appropriate alternative to hand hygiene. The hydrogel products of this product can also be used as "skin protectors", which can protect whole skin from various pathogenic materials as a preventive measure.

(23) In another preferred product version, silver-based products may be partially or completely replaced by silver / water compositions of the product. Silver EDTA (or AgEDTA) has been found to have antibacterial properties. Disodium

EDTA is a useful addition to the silver / water compositions of this product. However, ETDA (Ethylenediaminetetraacetic Acid) is an excellent synergist. EDTA (C10-H16-N2-O8) can be used in foods and is often added to alcoholic beverages as a preservative. EDTA is also used for heavy metal bonding therapy in humans. However, the use of AgEDTA as an antimicrobial agent (eg alone or in combination with other treatments such as those described herein) is not considered. The application of large markets, such as meat or protein production, the processing industry, the soap industry, the detergent industry (eg personal and household products), agriculture or the cereal industry and the healthcare industry, can all use stable silver, which can provide a variety of health benefits (both therapeutic and preventive). AgEDTA is easily accessible, and is easy to handle, store and transport. This version of the product indicates a new use of AgEDTA, that is, the use of AgEDTA powder in human, plant and / or animal care and in the treatment of various human and animal diseases (eg it can be used as a therapeutic or prophylactic treatment). AkzoNodel is currently manufacturing an approved AgEDTA. Other silver-forming or complementing agents, such as silver EDDS, silver cucrcuminate, silver berberine and silver tetracycline, have antibacterial properties and the use of these materials in the treatment of humans and animals is a new and previously unknown practice. Various other organic structures may be used to transport and / or transport silver and / or silver ions to different sites or biological structures. The amount of AgEDTA required will vary according to specific biological needs (eg treatment requirements or / and prevention).

(24) In another preferred embodiment of the product, additional silver-based inorganic products may partially or otherwise completely replace the silver / water composition. Silver (eg, silver ion, silver metal, Ag +) can be attached or attached, for example, to or between clay layers and / and zeolite cages. Such anchoring can occur by controlling, for example, the charge of the silicate layer, the charge of the zeolite cage, as well as the distance between the layers or the size of the zeolite cage. Silver can be attached or tied tightly. or rather liberated, it depends on the specific health applicable and the reaction between silver and biological properties (e.g., on the surface of the biological or internal portion or in the compounds of the interior, etc.). The goods may contain products which are liquid and drinkable or sprayable, as well as products in the form of a gel or ointment which may be sprayed on the surfaces as a gel or ointment.

(25) Each of the metals described herein can be stored in a crystalline or formless clathrate having one or more layers of atomic oxygen or oxygen-containing molecules. Certain metal / clathrate structures have shown excellent efficiency. In addition to the incorporation of silver and nets (eg zeolite) in the structure of the oxide layer (eg clays), silicates, phosphates and oxides such as hydrotalcite can also be used. Preferred clays from the mica group that can be used in the existing product (which can change the surface and / or spacing between layers) are: hydromuscovite, montmorillonite, chlorite and vermiculite.

(26) Clays or mica and zeolites are highly desirable as carriers of metal ions for many reasons, including many naturally occurring or simply obtained, and parts thereof may be retained in their desired colloidal size, such as suspended in liquids (eg water), and they are usually biologically harmless (no side effects or minimal). When silver is added to clay or zeolite chlorate, the molecules are heated to an average temperature (e.g., 100-200 ° C) to fix silver in chlorate. All of these materials can be created in varying degrees of viscosity, from liquid to highly viscous.

(27) Electron levels in an element such as a cation can be changed at any velocity position when the element cations are controlled by multiple anions. If the link is more covalent, several energy levels can be changed. Small or medium changes in the electron structure of silver may occur when silver is incorporated (or guided) into varying amounts of oxide ions. These changes in the electron structure of the cation, such as the silver cation, can occur in any structure of the various silver oxides. Silver cannot be placed in oxygen chloride or in any other way. Replacing, for example, the structure of the sodium cation with a silver cation may lead to the formation of sodium ion deposits in interchangeable cavities or spaces (eg on or between clay sheets or zeolite nets). The ability of a single material to change cations is called "CEC" or "cation exchange ability". CEC units are usually referred to as 'meq / 100 grams' or milliequivalents per 100 grams. The higher the CEC, the higher the ability of the material to accept cations (eg silver cations). Many oxygen-guided silver compounds can be used as carriers of silver (or other metals), so they can be used alone or combined with other treatments.

(28) Silver metals or silver ions contained in silica gel by diffusion or drying are the preferred technique for developing the metals of this study.

(29) In another preferred embodiment, the above-mentioned particulate compositions, organic and / or inorganic structures may be used to improve human and animal health. According to this product, the metal particles can be used separately or the metal particles can be combined with the organic compositions described above (AgEDTA). According to this product, metal ions can be combined with inorganic compounds (clays or zeolites). The metal ions of this product can be combined with both organic (AgEDTA) and inorganic (clay and zeolite) molecules. These delivery systems for silver metals or silver ions can be designed so that all consumption from any of the above silver delivery systems will provide silver to the body in varying doses. People can ingest a certain amount of silver through the mouth, internal organs, and through the large intestine, etc. Depending on the amount of clay or zeolite in water (as well as the various gel compounds described in this document), the end products of this study can be either liquid (low viscosity) or very viscous (high viscosity). If more clays and zeolites (including gel preparations) are added to the water, the final product will have a higher viscosity.

(30) Detailed description of the product, (31) This description is intended to ensure that qualified scientific personnel use the product and continue to use the intended best use of the product. Many of the modifications will remain because the basic principles of this product are defined in this document to form an improved silver / water composition (sometimes called silver nanoparticles dissolved in water) that can be used alone or blended (e.g. blended or supplemented) with other and which can form various hydrogel or ointment compounds that can kill human or animal pathogens both inside and outside the body.

(32) In essence, these products provide information on a new approach to kill or render harmless microorganisms that are dangerous to humans and / or animals. This approach involves the use of silver nanoparticles in water at a concentration of 5-40pmm silver, as well as the use of active silver particles containing, for example, AgEDTA or other compositions described herein. Depending on the application and / or the additives present, the silver / water composition can be used both internally and externally. Depending on the application, the silver / water composition may contain various desired admixtures, most of which are not noted herein, but will be apparent when additives with conventional chemical properties are used in the art.

(33) EXPANDED DESCRIPTION OF IMAGES.

(34) 1-6. The TEM microphotograph shows silver particles of different strengths taken in the silver / water composition, formed in accordance with this product.

(35) Figure 7a-7d shows TEM micrographs obtained from different TEMs using different techniques used in 1-6. fig. 7e. The figure below shows the EDS (EDAX) silver particle spectrum of this product's silver / water composition.

(36) Figure 8 shows a sample of electron diffraction obtained from silver particles in the composition of an existing article.

(37) Figure 9 includes three SEM photomicrographs showing the potential electron beam damage to silver particles obtained from the silver / water composition.

(38) Figure 10 shows SEM micrographs of a new silver electrode prior to use in this product.

(39) Figures 11, 12, 13 show dose 1, 2 and 3 shown in Fig. 10, initial EDS analyzes.

(40) Figure 14 shows a SEM micrograph of the tip of an electrode used to process the silver / water composition.

(41) Figures 15 and 16 show parts 1 and 2 of Fig. 14, initial EDS analyzes.

(42) Figure 17 shows a SEM photomicrograph taken from the tip of a 3500Χ usable silver electrode.

(43) Figures 18a and 18b are TEM photomicrographs of silver particles from the GNC Liquid Silver Supplement (25pmm).

(44) Figures 19a and 19b are TEM photomicrographs of silver particles obtained from the silver colloidal silver product.

(45) Figures 20a and 20b are TEM microphotographs of silver particles obtained from the colloidal silver product Vitamin World Bioorganic Advanced Collodial Mineral (3ppm).

(46) Figure 21 is a comparison of the TEM photomicrographs of five silver particles, two of which are derived from silver particles of this product and three are from commercially available colloidal silver particles.

(47) Figures 22a and 22b show seven different Raman spectra, 3 of which correspond to the silver / water compositions of the present product, one corresponds to pure water, one corresponds to deionized water and two corresponds to commercially available colloidal silver products.

(48) 23a. Figure 2 shows two Raman spectra corresponding to a patentable silver / water composition. Figure 23b shows three Raman spectra corresponding to three commercially available colloidal silver products.

(49) Figure 23c shows another Raman spectrum corresponding to silver / water compositions.

(50) Figure 24a shows the Raman spectrum of an existing product silver / water composition, Figure 24b shows the three Raman spectrum silver / water, zinc / water and copper / water compositions.

(51) Figure 25 shows a diagram of potential bacterial synergy diffusion test of the interaction disk.

(52) Figure 26 shows the titration of the structure and the graphs of the additive, synergy and anthogonal effects of the combination therapy.

(53) Figure 27 shows the sensitivity of isolated portions of MDR in lOppm silver compositions.

(54) Figure 28 shows photographs of antibiotic compounds of MRSA.

(55) Figure 29 shows photographs of antibiotic compounds of the E coil.

(56) Figure 30 shows photographs of Pseudomonas antibiotic compounds.

(57) Figure 31 shows a graph of the instantaneous power supply as well as the instantaneous silver concentration as a function of the process time during the formation of the silver / water composition. (58) Figure 32 shows an instant silver diagram as a function of process time using atomic absorption spectroscopy and electrical conductivity measurement techniques. The figure also shows the silver concentration 32 hours after production and after homogenization.

(59) Figure 33 shows an instantaneous current diagram, energy factor, and silver concentration as a function of process during the formation of a patentable silver / water composition.

(60) Figure 34 is a diagram showing SILDERM moisture loss.

(61) Figure 35 is a diagram showing the susceptibility of SILDERM to moisture.

(62) Figure 36 is a photograph showing the antibacterial activity of silver chelation (AgEDTA made by Akzo-Nobei) against pseudomonas fever (MDR).

(63) Figure 37 is a photograph showing the antibacterial activity of silver chelation (AgEDTA prepared by Alpha Chemical) against pseudomonas syphilis (MDR).

(64) Figure 38 is a photograph showing the sensitivity of SILDUST to the E-coil (MDR).

(65) Figure 39 is a graph showing the activity of SILDUST antiviral over time.

(66) Figure 40 is a photograph of a defined test plaque showing the formation of inflammation spots.

(67) Figure 41 is a photograph of a fixed assay plate which, after 3 hours, shows no inflammation spots, thus demonstrating the antibacterial efficacy of SILDUST.

(68) Figure 42 shows four X-ray diffraction properties of a patented silver / water composition; and four superimposed X-ray diffraction files (eg AgO, Ag2CO ₃ , Ag and Ag2O).

(69) Figure 43 shows the "TGA" Ag ₄ O ₄ analysis and "DTA" Ag ₄ O ₄ analysis.

(70) Figures 44a and 44b are SEM micrographs of a patented kaolinite / silver composition made in accordance with the present product.

(71) Figures 45a and 45b are EDS (EDAXX) analyzes corresponding to micrographs 44a and 44b.

(72) Figure 46 is a SEM micrograph of a new zeolite / silver composition made in accordance with the existing article.

(73) Figure 47 is a Linde 4A analysis of EDS (EDAX) zeolite containing the silver substituted here as well as prepared according to the present product.

(74) Figure 48a shows the UV-Vis spectrum of a 10 ppm silver / water solution and a 32 ppm silver / water solution greater than 190nm-400nm wavelength (prepared according to the current product). Figure 48b shows the same sample larger than the 190nm-250nm UV-Vis spectrum.

(75) Preferred product variants (76) Unlimited number of preferred product variants:

(77) A compound containing silver nanoparticles colloidally suspended in water with a total silver content between 5 and 40 ppm. This compound kills or renders harmless microorganisms which are hazardous to humans and animals (78) A compound containing silver particles, colloidally suspended in water, with a total silver content of approximately 10 ± 2 ppm. This compound kills or renders harmless microorganisms which are dangerous to humans and animals (79) A compound containing silver particles, colloidally suspended in water, with a total silver content of approximately 22 + 2 ppm. This compound kills or renders harmless microorganisms which are hazardous to humans and animals (80) A compound containing silver particles, colloidally suspended in water, having a total silver content of approximately 32 ± 3 ppm. This compound kills or renders harmless microorganisms which are hazardous to humans and animals (81) Hydrogel compound made of a particulate silver / water composition containing silver nanoparticles colloidally suspended in water with a desired silver content in the pre-particle material of about 32 ± 3. ppm (may be lower or higher). These hydrogel compounds kill or render harmless microorganisms that are dangerous to the human body. These compounds act as skin cleansers, wound healers and / or skin protectors as well as skin disinfectants.

(82) It should be noted that the material cannot be completely refined by determining the total amount of silver nanoparticles in the silver / water composition. Because the particles in the composition are tiny, the stated concentration of silver will have a high number of particles. The total surface area of the indicated silver concentration will increase. Therefore, particle size and particle size grades are important parameters for creating an effective patentable silver / water composition. Coatings such as oxide coating on silver particles (eg partially or fully coated) can increase the efficiency of the silver / water combination. Such coatings result from the processing conditions of the article. However, similar coatings of silver particles can be obtained by other processes (including non-silver metals such as zinc, copper, copper alloys, titanium, platinum and compound alloys) which are also discussed and associated with this product. When silver is applicable to the various additional metals described in this document, consideration should be given to their potential efficacy, which is dependent on certain biological conditions (eg particular pathogens involved).

(83) The further classification of variants is any of the compositions described above, wherein the maximum size of silver nanoparticles of more than 50% is less than 0.015 micrometers.

(84) The further classification of variants is any of the compositions described above, wherein more than 75% of the silver nanoparticles have a maximum size of less than 0.015 micrometers.

(85) The further classification of variants is any of the compositions described above, wherein more than 90% of the silver nanoparticles have a maximum size of less than 0.02 micrometers.

(86) The further classification of variants is any of the compositions described above, wherein more than 75% of the silver nanoparticles have a minimum size of less than 0.005 micrometers.

(87) The further classification of variants is any of the compositions described above, wherein the silver nanoparticles have a minimum size of less than 0.040 micrometers with more than 90% silver.

(88) The further classification of variants is any one of the compositions described above, wherein the silver nonparticulates contain a silver inert having a metallic oxidation degree (Ag (0)) in the core or central portion. At least one silver coverage ion oxidation selected from the group containing Ag (I), Ag (II) and Ag (III) with AGO, Ag2O / or Ag ₄ O ₄ coating must be a metallic silver core part (or the entire core).

• (89) The further classification of variants is any of the compositions described above, wherein the silver particles contain a silver inert metal having a metallic oxidation degree (Ag (0)) and a silver oxide coating with stoichiometry AgO or Ag ₂ O or other known stoichiometry. stable in the processing processes used to form the new silver / water composition of Ag ₂ O.

(90) Further experimental studies show that silver oxide coatings are present in at least part of the particles of the existing product and form, for example, Ag ₄ O ₄ - silver II oxide. In this molecule, two of the silver atoms may be 1 * (silver I), while the other two silver molecules may be 3 * (silver III). Under certain conditions, silver may be present in the 2 * (silver II) grade, resulting in at least a partial coating, such as Ag ₂ O. These coatings result from the processing conditions of an existing article (e.g., electrode / water interaction) and can affect overall the effectiveness of the silver / water composition. So far, it has been difficult to determine the exact composition of coatings, however, the description of the studies is described below.

(91) The further variant classification is any of the silver / water peroxide variants described above: the level of hydrogen peroxide in the final product is 1-3% by weight.

(92) The further variant classification is any of the silver / water variants containing. The final product contains 0.5-10 ppm disodium EDTA.

(93) The further classification of variants is any of the silver / water variants described above where about 50-75% by volume can be replaced by 10% povidone iodine. In the final product, iodine replaces about 25-50% of the silver / water composition.

(94) The further classification of variants is any of the silver / water variants described above containing various commercially available antibiotics (both liquid and powder) to form synergistically effective 'compounds'.

(95) The following classification of variants are methods of using all of the above compositions against a human or animal pathogen: (1) internally, (2) externally, (3) both internally and externally.

(96) The following classification of variants includes the use of AgEDTA to improve human or animal health.

(97) The further classification of variants includes the use of other silver products such as silver EDDS, silver curcumin, silver berberine and silver tetracycline.

(98) The further classification of variants includes the use of other metals such as zinc, copper, copper alloys, titanium, platinum and compound alloys, thus replacing silver in both the preparation and treatment methods described in this document. For the sake of brevity, silver predominates in this document, but it should be noted that other metals described in this document may be equally useful.

(99) In another preferred product variant, additional silver-based inorganic products may be partially or completely replaced by the silver / water compositions of the existing product. Silver (eg silver ion, silver metal, Ag +) can be added or attached, for example, between clay layers and / and in zeolite cages. Such anchoring can occur by controlling, for example, the charge of the silicate layer, the charge of the zeolite cage, as well as the distance between the layers or the size of the zeolite cage. Silver can be added or bound tightly or fairly liberated, it depends on the specific health applicable and the reaction between silver and biological properties (e.g., on the surface of the biological or inner part or in the joints, etc.). The goods may contain products which are liquid and drinkable or sprayable, as well as products in the form of a gel or ointment which may be sprayed on the surfaces as a gel or ointment. Each of the metals described herein can be stored in a crystalline or formless clathrate having one or more layers of atomic oxygen or oxygen-containing molecules. Certain metal / clathrate structures have shown excellent efficiency. In addition to incorporating silver and nets (eg zeolite) in the structure of the oxide layer (eg clay), silicates, phosphates and oxides such as hydrotalcite can also be used. Preferred clays from the mica group that can be used in the existing product (which can change the surface and / or spacing between layers) are: hydromuscovite, montmorillonite, chlorite and vermiculite.

(100) Clays or mica and zeolites are highly desirable carriers of metal ions for many reasons, including many naturally occurring or simply obtained, parts can be retained in their desired colloidal size, such as suspended in liquids (eg water) as well as they are usually biologically harmless (with little or no side effects). When silver is added to clay or zeolite chlorate, the molecules are heated to moderate temperatures (eg 100-200 ° C) to fix silver in chlorate. These materials can be created in varying degrees of viscosity, from liquid to highly viscous [100] Clay or mica, as well as zeolites, are highly desirable as metal ion carriers for a variety of reasons, such as many being naturally occurring or readily obtainable, particles can be stored in the required colloidal size which makes it possible, for example, to be used in suspension in liquids (such as water) and is usually very bio-friendly (ie with little or no side effects). Thus, when placing silver on or on, for example, clay or zeolite, the molecules are immediately heated to a moderate temperature (e.g., 100200 ° C) to attach silver to or inside the clathrate. All of these materials can be prepared in varying degrees of viscosity, from very liquid to very viscous.

[101] In addition, one of the invented preferred techniques for the release of metal ions is the incorporation of silver metal or silver ions into silicate gel by diffusion and drying.

[102] In another preferred modification possible in the inventive combinations of said particles, the organic and / or inorganic structures can be used to positively affect human and animal health and welfare. In particular, according to the invention, the metal particles can be used alone, as described above. Metal particles can also be combined, for example, with the organic compounds discussed above (eg AgEDTA). In addition, metal ions according to the present invention can be combined with any inorganic compounds (e.g., clays or zeolites). In addition, the metal ions of this invention can be combined with both organic molecules (e.g., AgEDTA) and inorganic molecules (e.g., clays or zeolites). This combination of silver metal or silver ion release systems may be designed such that, for example, the internal consumption of the above silver release systems may result in the release of silver at various sites such as the body. In particular, for example, people may ingest certain types of silver through the mouth, through the intestines, and through the large intestine and / or small intestine. Thus, depending on, for example, the amount of clay (s) or zeolite (s) associated with water (as well as the various gel-forming compounds listed here), the final product (s) may be very liquid (low viscosity) or very viscous (high viscosity). Thus, in general, the more clay or zeolite (as well as the gelling agent) associated with water, the more viscous the final product.

[103] EXAMPLES [104] COMPOSITION FORMATION [105] The silver / water composition can be formed according to the procedures provided in U.S. Pat. 6, 214, 299, the subject of which is specifically incorporated herein by reference.

[106] To prepare a composition composed of silver, according to the present invention, the preferred method is to use a chemical power source formed by the electrodes, and to do the following:

[107] (a) at least two high purity silver electrodes must be placed;

[108] (b) conducting a current through the silver electrodes, separating enough silver particles from said silver electrode to cause dissolved silver particles to form in the water, and [109] (c) stirring the water during said dissolved silver particles to disperse the dissolved silver particles. at a constant concentration in this water so as to produce a large quantity and a remarkably uniform distribution of dissolved silver particles in a single batch.

[110] In another preferred method, a chemical power source should be used to prepare the composition, which consists of silver / water compounds, and should do the following:

[111] (a) An electrical circuit shall be provided consisting of a power source to which the first conductor shall be electrically connected and the second conductor shall be electrically connected, the first conductor being at a fixed distance from the second conductor and at least one of the conductors is made of regular silver or zinc. copper and copper alloys, titanium, platinum and its alloys or alloys;

[112] (b) close the circuit by placing the first conductor and the second conductor in contact with a limiting resistor;

[113] (c) operate a power supply to simultaneously supply alternating current to the first conductor and the second conductor such that the voltage increases and decreases in the first and second conductors in a variable sequence, causing silver (or other metal) particles to separate from the first electrode; the suspension of this limiting resistor; and [114] (d) selectively arrange the electrodes against the barrier resistor to compensate for the reduction in electrode length due to the gradual separation of silver particles and to prevent curvature between the electrodes and the barrier resistor, and to maintain the desired current density at the ends of the electrodes.

[115] Each water chamber or reservoir producing silver / water components has a power supply consisting of eight transformers tuned to 120 vac incoming voltage (a modern invention that can be used as an acceptable transformer is Francformers (Franceformer)). No. 48765) and at a maximum output voltage of 10,500 vac at 30 milliamps. It was recommended that each transformer be equipped with a 45 microfarad capacitor (such as an aerovox, serial no. M24P3745MP2) which is connected in parallel to the power supply.

[116] The combination of transformer and capacitor may be useful in some cases and highly recommended in others. in special cases, the transformer assists with voltage supply and alternating current sine waves in a single phase. The degree at which voltage and current are present in relation to one another is the power factor. The closer the power factor to 1.0, the closer the volt and amper phase coincide and the more power delivered to the electrodes (eg power is usually determined by multiplying volts by amps).

[117] Each reservoir is provided with a transparent cover made, for example, of an appropriate polymer and is designed to receive eight sets of electrodes. Which set of electrodes consists of a fixed electrode made, for example, of an 18-size silver plate, and a plurality of usable electrodes arranged, for example, of 18-gauge silver wire (.9999 purity). The electrodes are preferably bent in the middle and result in twisting together in a double helix to obtain the desired combination of voltage and power density. Each set of electrodes is powered by one transformer.

[118] As each tank is intended for production, the electrodes are arranged in such a way that the fixed electrodes are in good contact with water (eg at least 1/3 to '/ ₂ plates are submerged) and the electrodes used are over water). When the power supply is charged, water rises and forms a cone-like structure around the electrodes used. This tapered structure is known in the literature as the "Taylor Cone." Initially, the water is very clean and has high electrical resistance. Accordingly, when using, for example, a constant current, 10,000-volt transformer, the corresponding voltage around the electrodes can initially be very high, such as 6500-8500 volts, and the electrodes used can be 5-10mm above water, thus achieving the desired voltage and desired current the strength of the electrodes used. This results in a relatively large Taylor cone for low water conductivity and high electrode conductivity (e.g., a wide field is created). The silver nanoparticle product is created when the silver particles have moved from the electrodes used to the surface of the air-water-silver electrode. As water absorbs more and more silver particles, the electrical resistance of the water / silver mixture decreases. Under constant or limited current, the relevant voltage will then drop or decrease as a function of time (see eg Fig. 31). According to this, the electrodes used are usually close to the water surface, e.g., probably 1-2 mm above the water. Simply put, Taylor cones will be much smaller due to the reduced difference in conductivity between water and electrodes (eg now a smaller field). Generally used electrodes and / or water levels during the manufacturing process should retain their initial geometry. Although Taylor cones remain smaller during the process (thus representing, for example, dissolution of metal particles), there will still be small Taylor cones at the end of the process. Water in each reservoir is charged throughout the process, thus maintaining homogeneity.

[119] Once the silver ppm silver / water solution has the desired amount or target, the product can be filtered, if necessary, with a 1 micron filter in any desired size, e.g., 2,300 - 6,500 gallon extra tanks and analyzed prior to loading product handling. The analysis is carried out by digestion using heat and acid and is performed using a PerkinElmer 300 Atomic Absorption Spectrophotometer. The resulting silver / water composition can then be combined with other ingredients to form a hydrogel, a sheet of material, or used in the same way, or combined with other additives (e.g., in any case as a liquid or dried and added as a powder) controversially.

[120] Referring again to Figure 31, which shows that real-time voltage drops and silver concentration data as a function of time correspondingly to the process of formation of the original silver / water components. It is clear that as the process progresses, the voltage decreases as the silver concentration increases. A consistent decrease in Taylor cone size is visible for each electrode. The silver concentration data in this graph should not be interpreted as quantitative, but as representative, prop and mixing data (eg, the silver / water mixture may not be completely homogeneous at all times when removing the prop).

[121] Referring now to Figure 32, which shows that there are two graphs of silver concentration in the same process, as well as different end results of the added concentration. The gray line with squares distinguishes the immediate concentration of silver (crucial for spectroscopic atomic absorption) on the basis of a 60 ml sample, pipetted from approximately the average depth of the reservoir and approximately midway between the center and the edge of the reservoir. The black line with the diamonds distinguishes the immediate silver concentration as a previously calibrated electrical resistance meter for 60 ml partial samples of the above liquid. As with the raw resistance data, at the beginning (i.e. time = zero), the water had a resistance of about 175 kilo centimeters. In contrast to the 31 hours of the process, the water / silver composition has a prestige of approximately 62.7 kilometers.

[122] There is one result immediately below the concentration / resistance result at the 32 hour mark marked as a square. This result shows the concentration of silver as a decisive factor in the absorption of the atoms by spectroscopy after high voltage setting but allowing the mixer to operate for another 20 hours to homogenize the composition.

[123] One of the conclusions from Figure 32 is that despite the operation of the mixer during the process, the silver may not initially be homogeneously distributed across the reservoir as it is formed. It can rarely be an accurate time interval after adding silver additives to the liquid and before the mixer captures and homogeneously distributes the silver in the water.

[124] Figure 33 is another graph of instantaneous voltage and silver concentration as a function of time during the silver / water production process. This graph also shows the instantaneous power factor of the power transformer. In this way, the power factor increases from 0.8 to a maximum of about 0.97 in 6 hours, ub decreases to 0.6 in about 30 hours. The voltage / time data were then mathematically equated to y = 2.1333 Ln (x) + 8.7057, where y corresponds to voltage and x to time. Silver ppm in water is shown in squares and starts at about 1 ppm and reaches a maximum of about 11 ppm after 30 hours (eg data if the water is not completely clean after filtration).

[125] PHYSICAL CHARACTERISTICS [126] Analysis of the silver content of the silver components of this invention can be performed by (acitylene) flame atomic absorption spectroscopy (FAAS), induction plasma (ICP), atomic emission spectroscopy (AES) or other techniques having one of the following: normal abilities, to be sensitive to silver at the appropriate concentration. If the silver composition particles are small and uniform in size (e.g., 0.01 micrometer or less), possible accurate analysis can be obtained by running on a colloid with atomic absorption or ICP / AES. This is because sample preparation for absorption of spectroscopic atoms essentially ionizes all silver, allowing it to be detected.

[127] If the components contain particles of 0.2 micrometre, the digestive method is recommended. The digestion process is not as perfect as the silver components need it to be, as it can form or persist upon contact with halides or other anionic agents which may react with the finally broken silver or bind to protein or other gel-like material. The process of digestion proceeds as follows:

[128] Take a 10 ml portion of the fully mixed or shaken silver composition for analysis and place it in a clean polycarbonate flask or other appropriate container (mainly a flask) with a well-fitting closure. The recommended size is 30-100 ml.

[129] Add 0.1 ml of nitric acid, using a micropipette or pipette, the silver composition in the flask.

[130] On leaving the flask in closed composition is heated to at least 80 ° and preferably about 90 ⁰ to 100 ° as long easy to stir up the silver is dissolved.

[131] Cool the resulting mixture to room temperature, leaving the flask closed. Shake well the flask. This digestion process also dissolves any silver oxide surface sediment that may be present in the silver particles.

[132] Use atomic absorption spectroscopy, ICP / AES, or similar to analyze the silver content of the silver composition. It is recommended to use a freshly prepared regulatory standard, preferably prepared according to the manufacturing inventory instructions that apply to the solution needs.

[133] If results are reported, one should collect all solutions at the time of preparation, including the 1% solution resulting from the addition of nitric acid.

[134] The silver concentration of the silver / water components in the present invention according to 31, 32, 33, et al. image data is determined using a Perkin Elmer AAnalyst 300 Atomic Absorption Spectrometer. Examples of the silver / water components produced are obtained according to the process described above.

[135] Principles [136] The Perkin Elmer Analytical System AAnalyst 300 incorporates a high efficiency burner system with a Universal GemTip nebulizer and an atomic absorption spectrometer. The burner system provides the thermal energy needed to dissociate the chemical compounds by providing free analytical atoms for atomic absorption. The spectrometer measures the amount of light absorbed at specific wavelengths using a hollow cathode lamp as the primary light source, as a monochromator and as a detector. The deuterium arc lamp adjusts the background absorption caused by the non-atomic species in the atomic cloud.

[137] PHYSICAL / CHEMICAL ANALYSIS OF SILVER AND SILVER / AQUEOUS COMPONENTS [138] Introduction [139] A composition sample, typically containing 22 ppm silver in water, has been analyzed in a process ion ion mass spectrometry (TOF-SIMS) to determine in the composition. The conclusion is that the amount of silver is in the form of silver (0) (that is, metallic silver) and that there is a layer of surface that is usually some component, such as silver (II) oxide (AgO). As mentioned above, silver (Π) oxide is usually a stoichiometric mixture of silver (I) and silver (III).

[140] Experimental Process B [141] Some drops of the inventive silver composition of 22 ppm have evaporated to dryness of the silicone substrate at ambient temperature. The precipitates were analyzed by TOF-SIMS and marked as an example. The silver (II) oxide (AgO) sample material was analyzed by placing a few particles of the sample powder, as collected, on a silicone substrate and labeled as a sample.

[142] Process Time Ion Mass Spectrometry (TOF-SLMS) technology is based on the principle that a solid sample is bombarded with an excited, oriented primary ion beam and then secondary ions emitted from the surface of the sample are analyzed for process time mass spectrography. This analytical technology relies on surface sensitivity to obtain information from a layer occurring at approximately 20 to 20 A (one Angstrom = 1x104 mirometers) below the surface. TOF-SIMS technologies are commonly used as a research mechanism to identify unknown sample compositions. Quantification is where possible relevant microanalytical standards are available for calibration. This assay was generated using standard high mass solubility conditions.

[143] C Results [144] A negative ion mass is used for the Ag (II) O material sample and the corresponding product sample. Mass spectral range of the solution spectra showing more than one representative of the silver oxide possibly present there as at least part of the silver particle layer. The data suggest that silver (II) on the surface of the silver sample particles has a moderate oxidation state.

Silver oxide (eg AgO) signals show a higher intensity in the first sample taken compared to the product sample, possibly due to the metallic dominance of the sample. This will be appreciated because as the average particle size in the sample decreases, the proportion of silver to silver oxide will also decrease and more silver oxide will be present.

[145] SIZE / MORPHOLOGY / COMPOSITION ANALYSIS [146] It is possible that the silver / water preparation efficiency described here is related to the particle surface properties (eg oxide / metal) and / or silver nanoparticle dispersion size and / or silver nanoparticle morphology. The smaller the average particle size, the larger the surface area and the greater the contribution of the particle surface chemistry. However, if the particles are too small, there may be loss of stability and / or other incidents that may adversely affect the product. The silver / water components of the instant product are noticeable because they are collectively present in pure water without active ingredients, etc. (e.g., various main types of colloids of silver make proteins retain their silver particles in suspension). Thus, silver / water compositions are essentially colorless, whereas other colloidal silver preparations (especially those with large particles) are generally colored. These properties are the result of production conditions, as analyzed above.

[147] Numerical analysis of the components showed an average particle diameter of 0.0106 micrometers, varying from 0.005 to 0.0851 micrometers. Whatever the size distribution, more than 95% of the particles have a diameter between 0.005 and 0.015 micrometres.

[148] Further elemental analysis was performed using SEM, EDA (EDAX) and TEM. In the particle, the silver / water compositions were dried and placed on an EM grid and studied using SEM (Scanning Electron Microscope) and two different TEM (Transmission Electron Microphones). These analytical tools determined the particle size distribution in the 10-30 nm range. However, microscope-generated images were needed to estimate the particle size because the particles tended to clump together or accumulate in the pile upon drying. The size of the dried agglomerates ranged from 50-100 nm. Figures 1-6 show various TEM images of dried silver elemental particles from the current invention silver / water connection. Figures 7a-7d show various TEM micro-images of silver elementary particles taken in accordance with the present invention, where these micro-images were generated using another technique. The silver / water compounds of this invention were placed on a C-film and examined using crio-TEM (a TEM other than the TEM used for numbers 1-6) at about -100 ° C. Thus, the silver / water connection was immediately frozen. Crio-TEM was used at about -100 ° C and about 100kV current, and the generated micro images are shown in Figures 7a, 7b and 7c. Figures 7a-7c clearly show that the average particle size is less than 20 nanometers. Additionally, Figure 7d shows a TEM analysis in SAD mode. Generally, these TEM micro images (Figures 7a-7c) show a maximum size of non-stick silver particles of 15 nanometers or less and some smaller particles in the range of 3.5-5 nanometers. The diffraction analysis in Figure 7d shows that the elemental particles are metallic silver, doubled in number and essentially pure. Figure 7e shows a silver particle taken from the silver / water connection of the present invention

EDAX spectrum (Energy Dispersion Spectrum or EDS). Figure 7e shows that silver does not contain any non-metallic slag. The copper present is from the necessary microscope equipment. There is evidence of a large amount of oxygen, which may be present in copper, as well as a silver particle region.

[149] Figure 8 shows a sample of electron dispersion taken from the silver elemental particles of the present invention. These data indicate the presence of at least one species of silver oxide. This data can be used for interpretation, but Figure 9 shows, for example, the potential damage to the electron beams caused by silver particles in the data collection process. This electron beam damage did not occur when researching colloidal silver developed by other manufacturers. So collecting data using SEM and TEM techniques is difficult because electron beam energy can cause damage. Therefore, these results were generated and analyzed with great care.

[150] Figure 42 shows the results of another tool. In this case, X-ray powder dispersion technique was used to prove the presence of oxide. Figure 42 shows four X-ray dispersion samples taken from four different dried 200 ppm silver / water junctions made in accordance with the present invention. The 32 ppm silver / water compound corresponding to this invention was concentrated up to about 200ppm using a standard reverse osmosis water filtration process. The silver / water connection was run through a reverse osmosis filtration system, where the excess water filtered contained much more silver elemental particles. As soon as the 200 ppm solution was obtained, the solution was dried under nitrogen to produce a powder to be investigated using X-ray dispersion. The silver / water connection was placed in a container, the container was covered with a plastic cover and nitrogen was introduced at one end and the nitrogen came out at the other end. The apparatus temperature did not exceed 75-80 ° C to maintain the integrity of all silver / water compound components. Subsequently, there was sufficient powder (obtained from the 200ppm solution) for X-ray dispersion analysis.

[151] Generated X-ray dispersion samples clearly show the presence of at least four separate types. In this regard, it is clear that silver carbonate clusters appear at 18-22 degrees. The presence of CO2 in the air was unavoidable, although efforts were made to develop a nitrogen blanket over the 200ppm solution. At 33 degrees, clusters of outbreaks appear. However, each of these outbreaks can be attributed to silver oxide (AgO)), silver carbonate (Ag2, CO3), and / or silver oxide (Ag2O). Thus, it is not clear which type is present. At 38 degrees, a strong outbreak of silver metal appears. This strong outbreak can be seen on each X-ray dispersion sample. It was found that silver oxide (Ag2O) flares appeared at 38 degrees. At 37 degrees, strong outbreaks of silver oxide (AgO) occur in combination with relatively strong outbreaks of silver carbonate (Ag2CO2). Silver oxide (AgO) outbreaks correspond to one of the silver oxide rectangular phases. By examining the generated X-ray dispersion data and comparing it with existing database files, it was determined that silver / water compounds contain one or more silver oxide phases. It was noted that no X-ray dispersion samples were available to compare Ag4O4 with the X-ray dispersion samples of this invention.

[152] However, Ag4O4 does not exist commercially. In this regard, a sample of Ag4O4 was obtained and TGA and DTA analysis of such powder was performed. Figure 43 shows TGA and DTA analysis. The DTA curve in Figure 43 indicates that the Ag4O4 endotherm exists at 181 ° C. This endotherm corresponds to the weight loss shown in the TGA curve in Figure 43. These experimental characteristics are consistent with Ag2O4 degradation in Ag2O. Another very strong endotherm is seen at 403 ° C, as well as a second weight loss. These two esoteric values correspond to Ag2O decomposition in Ag metal.

[153] Figure 10 shows a SEM micro-image of a new silver electrode before it was used in a process according to the present invention. The EDS element analysis was performed on the electrodes designated 1,2 and 3. These analyzes are shown in Figures 11, 12 and 13. These analyzes showed the presence of pure silver.

[154] Figure 14 shows SEM micro-images of the silver electrode tip used after its use in the process of this invention. The EDS element analysis was performed on the electrodes used, designated 1 and 2. These two separate analyzes are shown in Figures 15 and 16. Figure 17 shows enlarged (3500 times) SEM micro images of the electrode tips used. Portions 4 and 5 were also tested by EDS element analysis and were also found to contain pure silver.

[155] Comparison of Commercially Available Colloidal Silver Elemental Particles [156] To understand the differences in the realization of silver / water compounds (e.g., bioscale effect) compared to known colloidal silver, physical properties were tested. Figure 18a and 18b are TEM flasks of silver particles corresponding to silver particles in the first colloidal silver obtained from the General Nutrition Center in 2004 and known on the market as GBC Liquid Colloidal Silver Additive (25 ppm) (GNC). Figure 19a and 19b are TEM micro-images of silver particles corresponding to the second colloidal silver known on the market as "Silverado". Figure 20a and 20b are TEM micro-images of silver particles corresponding to the third colloidal silver known on the market as the "Vitamin World Bioorhanic Advanced Colliodal Mineral" (3ppm) (organic). Figure 21 is a TEM micrograph of the silver particles of two silver / water compounds (designated "ASAP 2" and "ASAP 10") and the three above-mentioned colloidal silver coatings. These images show a clear difference in particle size and shape, thus showing that there are physical, structural and possibly chemical differences between the different colloidal silver, which partly explains the biological differences between the different products.

[157] Spectroscopy Characteristics [158] RAMAN Spectroscopy [159] Further analysis of silver / water mixtures was performed using Raman spectroscopy. Several analytical approaches were made using three different Raman spectroscopes. The reason for using the Raman and Resonance Raman spectroscope was that different vibration modes (and / or amplitudes) could be trapped in different colloidal silver as compared to the silver / water compounds of this invention, as well as to "pure" or dionized water. These different modes of vibration in water molecules can help to better define colloidal systems and biological differences in different silver products.

[160] For the first Raman spectroscopy measurements, a Confocal Raman microscope from Vitech (Ulm, Germany) was used. The model number was CRM200. Spectra were obtained using a Nikon 60x immersion lens (NA = 1) with a spectral integration time of 15 seconds (three separate 5 second acquisition times). A drop of liquid was placed in the small dish well and an immersion lens was inserted into it. The laser source was 532nm with about 10mW. A confocal detection system was used with a confocal volume of about 0.3 x 0.3 x 0.75 micrometers (about 7x10 E-8 picometers).

[161] Figure 22a and 22b are graphical results of the data collected for the seven samples. Two of the samples are the same, albeit differently labeled (10PR and 10 PSU), and correspond to the above "ASAP 10" (10 ppm silver from silver / water connection). "HPLC" corresponds to high purity (ultra purity HPLC) water from Alfa Aesar. "DL" corresponds to dionized water. "GNC" corresponds to GNC Liquid Colloidal Silver Additive (25 ppm). The AGX-32 corresponds to a 32 ppm silver / water connection. VW corresponds to Vitamin World Bioorganic Advanced Colloidal Mineral (3 ppm) (formerly Bioorganic). A clear difference is shown between the different samples. For example, the primary tensile regime (approximately 3400-3500 l / cm) exhibits large differences in these few water / water based compounds. Vibrant / rational behavior below 500 1 / cm also shows clear differences between samples. Some differences can also be seen in the bending mode of about 1600 1 / cm.

[162] The second Raman dataset was generated from different spectrograph systems. The numbers generated between the two data sets are different (hence the Raman spectroscope data for water is a function of the analytical device used) and the data in this data set also show significant differences between the silver / water compounds of this invention as compared to other colloidal silver or water. A Raman reflection microscope was used in this set of Raman spectroscopy measurements. Spectra were obtained using an Olympus 20x lens (NA = 4). The CCD detector was centered at 1600, 2500, 3400 and 4400 1 / cm. The laser source was 514.5 nm with approximately 11.5 mW. Further spectrum information can be found in Figures 23a, 23b and 23c. The sample designation in these images corresponds to the previous text. Both Raman spectroscopy data sets indicate that these different samples exhibit varying molecular motions indicating the biological activity of silver / water compounds according to the present invention.

[163] A third Raman dataset was generated using a third multi-laser micro spectrograph by Renishaw Confocal Raman. This system was configured to allow measurements above and below the sample. The design was designed to investigate a sample size 100x to 1000x larger than that described in the first set of measurements. The reflection micro spectrograph with a Leica DL DM microscope was immersed in 20x (NA = 0.5) water immersion or 5x (NA = 12) in a dry lens. Two laser frequencies were used - a multi-line 50mW Argon laser with / 2 current for 514.5nm and a 20mW HeNe laser for 633nm. High-resolution gratings were inserted into the optical passageway of the monochrometer, allowing continuous scanning from 50-4000 (1 / cm). Integration times of 10 to 20 seconds were used. The sample liquid was placed under a lens in a 50ml beaker. Both lasers were used to study the resonance compounds, while the second laser was used primarily to obtain the Raman spectrum. The sample size was about 25ml. Measurements made with a 5x dry lens were made with an object placed 5mm above the liquid to examine a volume of about 7mm below the water meniscus. Immersion measurements were made with a 20x immersion lens placed in a sample of approximately 4mm to allow for the same volume of study. The CCD detector acquisition areas were individually adjusted for each lens to increase signal intensity and signal-to-noise ratio. An example of the silver / water bond spectrum of this invention is shown in Figure 24a. Figure 24b shows the Raman spectrum of three different metal / aqueous fluids; area 2 corresponds to 10ppm zinc / aqueous liquid; and area 3 corresponds to 1ppm of copper / water liquid.

[164] The number generated between the two data sets differs (hence the Raman spectroscope data for water is a function of the analytical device used and the device design) and the data in this data set also show significant differences between the silver / water compounds of this invention compared to other colloidal silver or waters. All Raman spectroscopy data sets indicate that these different samples have different molecular motions and compounds, thus demonstrating the effectiveness of the silver / water compound. Differences between Raman samples in the three different metal / aqueous fluids shown in Figure 24b also indicate various differences in exposure.

[163] UV-VIS SPECTROSCOPY [164] Further analysis of the silver / water compound was performed using a UV-Vis spectroscope. UV-Vis spectroscopy was used in addition to Raman spectroscopy to look for additional resolution modes and / or vibration amplitudes in different parts of the spectrum. One UV-Vis spectrometer was used to collect data. In this regard, the energy absorption spectrum was obtained using UV-Vis micro spectrometry. This information was obtained using a dual beam scanning mono chronometer system cable with a wavelength of about 190nm to IlOOnm. The UV-Vis spectrometer used to obtain the absorption spectrum was the Jasco MSV350. The instrument was installed to support measurements of low concentration liquid samples using a 10mm x 10mm fuzzy quartz tube. Data was acquired at this wavelength using a photo multiplier tube (PMT) and a photo diode detector with the following operating characteristics: 2nm bandwidth;

0.5nm resolution. In this regard, the UV-Vis signature for pure water was extracted from the generated spectrum to display more prominent spectral signatures of the silver / water compound.

[165] Both tungsten "halogen" and hydrogen "D2" energy sources were used as the primary energy source MSV350. The passage of the optical spectrometer was installed to allow the energy beam to pass through the samples to the center of the specimen clips. During specimen preparation, filling and covering of the straps was limited, as well as placement in the strap holder in a fully sealed specimen compartment. The information output was measured and represented as Absorption Units versus Wavelength and Frequency. The main difference between the samples corresponding to the two spectra (see Figures 48a and 48b) was the silver concentration of each sample. Specifically, the highest amplitude curve in each of Figures 48a and 48b corresponds to a 32 ppm silver / water solution; the lower amplitude curve corresponds to a 10 ppm silver / water solution. The wavelengths of the highest points of the frequency (i.e. the arrangement of the highest and the lowest points) and the positions are quite similar.

[166] As mentioned above, silver (eg, silver ions, silver metals, Ag +, etc.) can be added or fixed, for example, between and / or clay layers and / or in a zeolite cage. One method of obtaining placement, such as silver ions inside or on clays, mica or zeolite, is to provide the silver ion species in a soluble state and to introduce these species into the clay or zeolite composition. The concept of metabolism, for example silver ion, is sometimes referred to as "BEC" or "CEC" (both of which are shorthand nomenclatures for the corresponding "system" cation exchange capacity). In this regard, most kaolinite materials undergo a cation exchange with a capacity range of 2-5 (e.g. 2-5 meq / 100 grams). For example, montmoriolite clays have a cation exchange capacity of about 100meq / 100 grams. In contrast, the zeolite has a cation exchange capacity of up to several hundred meq / 100 grams according to BEC or CEC. In general, the more BEC or CEC, the higher the material's ability to receive cations.

[167] Experiments were conducted to determine the ability of cations or zeolites to convert to silver (or other metal cation (s)). The following steps were taken to prepare and thus analyze silver-clay samples as well as silver-zeolite samples.

[168] Thus, first, the conventional kaolinite and zeolite Linde 4A materials, which were washed three times with deionized water, were taken to eliminate the possible presence of chlorine contamination, which in turn could lead to the deposition (or undesirable) of certain silver materials (eg silver ions). reaction) before the silver material adheres to / inside the kaolinite and / or zeolite structures. These washed materials were then mixed with a solution of silver nitrate (AgNCn) in the appropriate concentration according to the desired or known "CEC" of each respective material. The materials so treated were repeatedly washed in deionized water to remove unused silver nitrate. Samples were left to dry overnight in an electrical resistance dryer at 120 ° C. The washing procedure is as follows:

[169] About two grams of each kaolinite or zeolite sample were placed in a centrifuge tube. Then deionizing water was added. Mix the sample and deionizing water for approximately 40 minutes. The compound was centrifuged for about 30 minutes at 1000 RPM. Excess liquid is drained from the pipe. Adding deionizing water, mixing, centrifuging and draining excess water is repeated three times in total.

[170] Silver nitrate is introduced into the purified kaolinite and zeolite materials after appropriate washing of the sample to remove any chlorine contamination. About 0.09 grams of silver nitrate is injected into the kaolinite solution, while about 4.25 grams of AgNO ₃ are injected into the Linde 4A zeolite. The calculated amount of silver nitrate is added to each tube, then the tube is filled with deionized water and stirred for about 40 minutes, followed by centrifugation for about 30 minutes at 1000 RPM. The excess fluid is then drained off. Adding deionizing water, mixing, centrifuging and draining excess water is repeated three times in total. After the washing and silver nitrate injection procedures are completed, the samples are removed from the centrifuge tube and placed in an aluminum (AI3O3) crucible and left to dry overnight in an electric resistance oven at 120 ° C. The resulting kaolinite / silver and zeolite / silver materials are characterized by SEM photomicrograph and SEM EDS (EDAX) methods. Figures 44a and 44b show SEM photomicrographs of kaolinite samples prepared according to the above methods. According to these photomicrographs, it can be observed that the book-like or leaf-like kaolinite structures (such as S1O2 and AIO2O3, indicated as "X" and "Y" in Figure 44a) clearly show that silver cations are located around the edges of clay materials. It is clear that some form of silver attachment or exchange has occurred, as evidence, the brightest leaf-like portions of this photomicrograph (Note: in this sample, the "X" and "Y" portions represent various other book-like structures). Figures 45a to 45b show the analysis of EDS (EDAX) samples shown in Figures 45a and 45b, respectively. These analyzes demonstrate the presence of aluminum and silicone, which is acceptable for kaolinite as well as titanium (rutile is desirable). It is also possible to see small silver highs corresponding to the BEC of kaolin, which is relatively low at 2-5.

[171] Figure 46 shows a SEM photomicrograph of a treated zeolite treated according to the above procedures. At higher zeolite CECs (about 500), the zeolite cube-like structure (Figure 46) looks like a photomicrograph, flaming ”(see Part A in Figure 46). This "glittering" suggests that the zeolite had a prominent uniform distribution of silver. In this regard, if the bright spots of silver that produce this glow are visible, then silver has not penetrated into the zeolite. Figure 47 shows an EDS (EDAX) analysis for the example shown in Figure 46. Relatively high amplitude points for aluminum and silicone appear, while silver peaks are particularly high (for example, compared to the Ag points in kaolinite, see Figures 45a 45b). These high silver points correspond to the greater ability of the zeolite to retain silver (e.g., high BEC) in its structure relative to the kaolinite structure (e.g., low BEC); 44a and 44b.

[172] Proof of 22 PPM SILVER COMPOSITION AGAINST

BACILLUS SUBTILIS [173] A. Purpose of Example

V [174] The purpose of this example is to demonstrate the antimicrobial activity of silver in relation to the present invention of bacterial spores based on the test organism Bacillus subtilis. Bacterial spores are generally not capable of eradication.

[175] Materials and Methods.

t [176] Test organism. A test suspension containing spores from Bacillus subtilis (ATTC # 19659) was prepared from a culture supplemented with additional spore enhancers. The plates were overlaid with sterile water and the spores were sprayed with suspension water. The final wash was done in 70% methanol for 30 minutes to make sure all bacteria were eliminated. The spores were rinsed in water containing 0.01% Tween 80 to prevent sintering.

Neutralized] s. Neutralizing composition: 12.7% Tween® 80, 6.0% Tamol® SN, 1.7% lecithin, 1% peptone and 0.1% cystine. This mixture is designed to neutralize any chemicals that may interfere with bacterial growth.

[177] Procedure for destruction:

[178] a) 9.9 ml of disinfectant (22 ppm silver compositions in water) were placed in a sterile 20 mm x 150 mm tube. The tube is placed in a 20 ° C water tank.

[179] b) 9.9 ml of disinfectant (22 ppm silver compositions in water) were placed in a sterile 20 mm x 150 mm tube. The tube is placed in a 20 ° C water tank.

[180] (c) After 30 min, 1 h and 4 h, one ml of the organism / disinfectant was poured into a tube containing nine ml of neutralizer. The tube was shaken.

[181] d) After two minutes, the neutralizer suspension was diluted 1:10 in saline (PSS).

[182] e) The number of viable organisms in the selected dilution tubes was tested by membrane filtration. One ml of solution was placed in a double. The membranes were washed in 100 ml sterile PSS and placed on plates. The plates were heated at 37 ° C for 20 h.

[183] (f) Colonies were counted and thoroughly processed on each filter.

[184] Monitoring:

[185] a) Test suspension titers (unit) were calculated by membrane filtration assay at the selected 1:10 test suspension solution PSS.

[186] (b) Neutralized monitoring was performed by administering a composition consisting of 9 ml of neutralized and 1 ml of disinfectant together with 100 ml of titre solution containing 100 cfu. This produces about 10 cfu / ml in the tube, which was left for 20 min before testing the membrane filter using a double 1 ml sample.

[187] C. Results [188] Bacillus subtilis titers (unit):

Solution n:

l: lx ιοί 1: 1x10 7th 1: 1 x10 8th TNT Number of colonies: 75 7th TNT 58 8th

[189] TNTC = Too Many to Add

Dilution of Bacillus subtilis spore / disinfectant suspension:

Time 1: 1x10 ' l: lx10 ² l: lx10 ³ l: lx10 ⁴ l: lx10 ⁵ l: lx10 ⁶ 30 min - - TNTC TNTC 57 10th - - TNTC TNTC 57 7th 1 hr - - TNTC TNTC 28th 3 - - TNTC TNTC 55 3 2h - TNTC TNTC 126 23rd - - TNTC TNTC 183 17th - 4h TNTC TNTC 88 12th - - TNTC TNTC 69 12th - -

[190] TNTC = Too Large to Count [191] Neutralization Control: 1: 1x10 ^B [192] D. Discussion [193]. The results show the concentration of viable spores of B. Subtilis 6.65x10 ⁶ 5 spores per ml in the original suspension. Inoculation of 9.9 ml disinfectant with 100 ml of this suspension produces an initial concentration of 6.65x10 ⁶ spores per ml in the test tube.

[194] The results of these procedures allow the calculation of registry reduction (LR) and interest destruction (PK) to be calculated. These are summarized in the table below. Values were calculated using the formula: LR = -Log (S / So) and PK = (1- (S / So)) x 100; where S = concentration of organisms in a given time; and So = the initial concentration of the organisms at zero.

Time Reduction of the register Destruction of interest 30 min 0.090 18.8 lh 0.205 37.6 2h 0.634 76.8

4h '1.928 98.8 [195] Neutralization control data show that the disinfectant was adequately neutralized. Current calculations show that the result of the dilution is possible without appreciable destruction.

[196] disinfectant preparation has been tested in the page shown here for sporadic activity against B. Subtilis spores, a common species used in testing and belonging to the same group of genes as the organism causing anthrax. Due to the genetic similarities between the two, B. subtilus spores are subjected to non-pathogenic surrogate bacillus for the anthrax bacterium, anthrax.

In addition, these results are applicable to the Siberian plague.

Prolonged exposure is expected to result in additional destruction.

[197] EVIDENCE FOR COMPOSITION OF 10 PPM SILVER AND 1.0% H ₂ O ₂ AND 14 PPM SILVER AND 1.5% H ₂ O ₂

COMPOUND EFFICIENCY COMPOSITIONS [198] A. Reason for Example [199] The reason for the example is to demonstrate the antimicrobial activity of two compositions based on the present inventions of bacterial endospores from the test organism Bacillus subtilis. This was done using standard kill time assays using a suspension of B. Subtilis endospores.

Looking at a similar example to the previous example (using 22 ppm silver), this example demonstrates the potency of the hydrogen peroxide H ₂ O ₂ on the antimicrobial properties of silver compositions. Hydrogen peroxide is stable in the presence of silver compositions in the present inventions. As long as hydrogen peroxide has antimicrobial properties, it can be destroyed by catalytic or microbial enzymes. Although hydrogen peroxide is capable of weakening bacterial cell walls and providing silver particles before any enzymatic destruction that can occur with hydrogen peroxide. preparation of disinfectant has been tested on the page shown here for sporadic activity against B. Subtilis spores, a common species used in testing and belonging to the same group of genes as the organism causing anthrax. Because of the genetic similarities between the two, B. subtilus spores are subjected to non-pathogenic surrogate bacilli for the anthrax bacterium, the anthrax bacterium.

[200] B. Materials and Methods [201] 1. Testing the organism. The suspension to be tested contains Bacillus subtilis (ATCC # 19659) endospores prepared on nutrient Agar where additional sporadic enhancers were added. Plates were treated with sterile water and endospores were purified in multiple separator water. The last wash was 70% ethanol for 30 min to confirm the death of the vegetative bacterium. Spores were diluted in water containing 0.1% Tween 80 (polysorbate mark) to prevent adhesion.

[202] 2. The neutralizer. Neutralization mix containing 12.7% Tvveen 80, 6.0% Tamol SN (trace sodium naphthalene condensate), 1.7% lecithin, 1% peptone and 0.1% cystine. This solution was designed to neutralize any chemicals so as not to affect the growth of the bacterium. 30 [203] 3. Destruction Time Procedure:

[204] a) 9.9 ml of non-residual each disinfectant (colloidal silver inventive composition: one containing 14 ppm silver and 1.5% H2O2; the other containing 10 ppm silver and 1.0% H2O2) was placed in a sterile 20 mm x 150 mm tube. The tubes were equilibrated in a 20 C water bath.

[205] (b) Each disinfectant tube was inoculated with 100 ml of the test organism suspension at time zero.

[206] c) After 10 min, 30 min, lh, 2h, 4h, 6h and 8h, each 100 ml of test organism / disinfectant suspension was placed in a tube containing nine ml of neutralizer. The tube was MIXED THROUGH IT.

[207] D) After two minutes, the neutralization suspension was serially diluted 1:10 in physiological saline (PSS).

[208] e) The number of viable organisms in selected diluent tubes was tested by membrane filtration method. One ml without residual dividing was silver plated in double. Membranes were washed with approximately 100 ml sterile PSS and transferred to Columbia Agar plates.

Plates were incubated at 37 ° C for 20 h.

[209] (f) The number of colonies per filter was counted and the register reduction calculated.

[214] 4. Checks:

[211] a) Test suspension tubes were calculated using membrane filtration assay at a selected 1:10 ratio of test suspension

PSS.

[212] b) Control of the neutralizer was performed by inoculating 9 ml of the mixture with the neutralizer and 1 ml of disinfectant with 100 ml of l: lx10 ³ dilution tube. It produced approximately 2,000 cfu / ml in a tube held for 20 min at 1:10. Both tubes were tested by membrane filtration using [25] for 1 ml of duplicate samples. All results are shown in tables la and lb.

[213] Results:

[214] Spore tube of Bacillus subtilis:

Relationship

1: 1x10 'l: lxlQ ⁷ l: lxlQ ⁸ Colony count: TNTC 36 5

TNTC 27 4 [215] TNTC = Too Large to Count.

Table la

Solution containing 14ppm silver and 1.5% H2o2:

B. subtle spore / disinfectant suspension dilution

Time 1: 1x10 ' l: lx10 ² l: lx10 ³ l: Ix10 ⁴ l: lx10 ⁵ 10 min - - TNTC TNTC 227 - - TNTC TNTC 265 30 min - - TNTC TNTC 258 - - TNTC TNTC 273 lh - - TNTC TNTC 55 - - TNTC TNTC 33 2h - TNTC 207 29th - - TNTC 237 24 - 4h 59 3 1 .57 5 1 6h 0 0 0 3 0 0 8h 1 0 0 1 0 0

[216] TNTC = Too Large to Count.

[217] Neutralization control:

Undiluted: 1: 1x1o ¹

TNTC 195

TNTC 210 [218] TNTC = Too large to count.

Table lb

Solution containing lOppm silver and 1.0% H2o2:

B. subtle spore / disinfectant suspension dilution

Time 1: 1x10 ' l: lxl0 ² l: lxl0 ³ l: lx10 ⁴ 1: 1x10 ' 10 min - TNTC TNTC 230 - INTC TNTC 287 30 min - TNTC TNTC 254 - TNTC TNTC 260 lh - TNTC TNTC 146 - TNTC TNTC 124 2h TNTC TNTC 64 TNTC TNTC 71z - 4h TNTC 72 5 TNTC 77 5 6h 0 0 0 2 0 0 8h 0 0 0 0 0 0

[219] TNTC = Too Large to Count.

[220] Neutralization control:

Undiluted:

1: 1x10 'TNTC 200

TNTC 184 [221] TNTC = Too Large to Count.

[222] D. Discussion [223] The data represent a viable concentration of 2.59x10 ⁵ spores per ml of viable B.subtilus spores in the original suspension. Inoculation of 9.9 ml disinfectant with 100 ml of this suspension produced 2.59x10 ⁵ spores per ml in a test tube at an initial concentration.

[224] The results of these procedures allow the calculation of the ratio reduction (RR) and the deadweight percentage (AP). These are listed in the table below. The values are obtained using the following formula: RR = Log (S / So) and AP = (1 - (S / So) * 100), where S = concentration of organisms at a given time and So = initial concentration of organisms at time = 0. No significant mortality was observed within 30 minutes, and 10 minutes were applied to So values, whereas at 6 and 8 hours, the numbers were not used, and no linear regressions were used. The "fitted line plots" command in the statistical program "Minitab" The regression calculations and the time required for reduction are shown in Table 2.

Table 2

14 ppm SUDRABS + 1.5% H ₂ O ₂ 10 ppm SUDRABS + 1.0% H ₂ O ₂ Time specific weight reduction Percent Lost rating specific weight reduction Percent Lost rating 30 min -0.03 -7.9 0.003 0.6 lh 0.66 78.0 0.28 47.8 2h 2.05 99.1 1.58 97.4 4h 4.63 99,998 3.54 99.97

[225] Regression Analysis [226] Equation for 14 ppm calculation: Y = -0.66704 + 1, 32936x. Equation for 10 ppm calculation: Y = -0.59690 + 1.03933x. These equations show that the time required for six-fold reduction is 5.02 hours for the 14 ppm composition and 6.35 hours for the 10 ppm composition.

[227] Neutralization control data indicates that the disinfectant has been properly neutralized. The calculated calculations were consistent with those expected from the solution.

[228] The experimental solutions showed significant sporicidal activity against B. subtilis spores during the tests. The B. subtilis foci used in these studies were also used in AOAC sporicide assays. The spores of this organism pose a major challenge to most disinfectants. The time required for six-fold reduction goes hand in hand with the sporicide labeled requirements for many cold sterilizers.

[229] EXAMPLE OF THE EFFECTIVENESS OF 10 PPM SILVER COMPOSITIONS AS A BROAD SPECTACLE ANTI-MICROBUSIC [230] A. Methods [231] The MNK (minimum inhibitory concentration) and MBK (minimum bacteriological concentration) assays were performed according to the standard microscope. MNC is defined as the lowest level of antibiotic concentration (in vitro) that inhibits the development of an infectious organism. The results are expressed in micro grams per ml. The interpretation of in vitro data obtained with medical antibiotics is based on the achievable serum concentrations in the drug, which may vary with dose, route of administration, degree of binding protein, site of infection, age and weight of the patient, and a number of other factors. MBK is defined as the lowest level of antimicrobial agent needed to suppress 99.9% of the body's initial inoculum.

[232] The test was performed by culturing pure cultures of each test organism in a liquid environment. Turbidometric measurements were used to control the culture concentration level. Several nutrient compositions with antibiotics were used in each assay. The solutions were supposed to affect the sensitivity of the organisms. Each tube was injected with a known amount of test culture and allowed to incubate at 37 ° C (± 2 ° C). The tubes were tested turbidometrically for bacterial growth. At concentrations below MNK, the tubes exhibited optically thickening, indicating bacterial growth. Tubes that did not undergo growth processes were developed in a fresh environment. The test tube with the lowest concentration of antibiotics that did not undergo growth processes was adopted as MBK. The results are shown in Table 3.

[233] Results in Table 3 la

The organism Tetracyclin e Ofloxacin Penicillil G Cefaperazo n Erythrmyci n Silver S. pyogenes 0.625 /> 5 1.25 / 2.5 > 5.0 0.313 / 1.25 0.003 / 0.019 2.5 / 5.0 S. mutans 0.625 /> 5 2.5 /> 5.0 0.521 /> 5 1.25 /> 5 0.009 / 0.019 2.5 / 10.0 S. gordonii 0.156 / 0.625 2.5 / 5.0 0.009 / 0.03 9th 1.25 / 1.25 0.005 / 0.019 2: 5 / 10.0 S. pneumoniae 0.078 / 0.625 2.5 / 2.5 0.19 / 0.019 0.313 / 0.313 0.002 / 0.004 2.5 / 2.5 S. faecalis 0.313 /> 5 1.25 / 5.0 5.0 /> 5.0 > 5.0 0.09 / 1.25 10.0 / 10. 0 S. aureus 0.313 /> 5 0.417 / 0.62 5 2.5 /> 5.0 5.0 / 5.0 0.039 /> 5.0 5.0 / 5.0 P. aeruginosa 0.078 / 5 0.156 / 0.31 3 0.13 /> 5.0 2.5 / 5.0 2.5 /> 5.0 1.67 / 5 E. coli 1.67 /> 5 0.104 / 0.15 6th > 5.0 0.625 /> 5.0 5.0 /> 5.0 2.5 / 2.5 E. aeroganes > 5 0.078 / 0.15 6th > 5.0 2.92 /> 5.0 > 5.0 2.5 / 2.5 E. cloacae 1.67 /> 5 0.156 / 0.15 > 5.0 > 5.0 > 5.0 2.5 / 5.0

6th s. typhimuriu m 1.25 /> 5 0.078 / 0.15 6th > 5.0 1.25 / 2.5 5.0 /> 5.0 2.5 / 5.0 S. arizona 0.625 /> 5 0.078 / 0.07 8th > 5.0 0.833 /> 5.0 4.17 /> 5.0 2.5 / 5.0 S. boydii 1.25 /> 5 0.078 / 0.15 6th > 5.0 0.625 / 0.625 5.0 /> 5.0 1.25 / 1.2 5 Q. pneumoniae 2.5 / 5 0.417 / 0.62 5 > 5.0 > 5.0 > 5.0 2.5 / 2.5 K. oxytoca 1.25 /> 5 1.104 / 0.15 6th > 5.0 1.25 /> 5.0 > 5.0 1.25 / 1.2 5

[234] Data shown are MIC / MBC (Minimum Inhibitor Concentration / Bacterial Concentration of Mimmachah) per million parts (ppm); ">" Means that the concentration required to achieve the MIC or MBC is higher than the test parameters measured for the test case. For example, the highest concentration of teracycline used for S. piogenin was 5 ppm. At this concentration, subculture from the tubes "without growth" was still observed. Therefore, the MBC should be> (greater) than 5 ppm.

[235] MIC / MBC E.coli lineage 0 157: H7 associated with bleeding diarrhea and colitis was identified in the following studies. MICs were set at 2.5 ppm and MBC at 5 ppm.

[236] C. Conclusion.

[237] The 10 ppm silver composite was tested in this invention and found to be both bactericidal and bactericidal to all organisms tested. In other studies, this composite was compared with other commercially available colloidal silver products and its hyperactivity was compared with other tested formulations (data not shown).

15. The most interesting observation was the wide range of 10 ppm silver composite. The antimicrobial activity observed was almost constant irrespective of the type of organism tested. Except for Streptocococcus faecalis and Streptococcus aureaus (which have MIC values of 10 ppm and 5 ppm, respectively), MIC values were between 1.25 and 2.5 ppm for both gram positive and gram negative organisms. MBC values behaved similarly to values from 1.25 ppm to 5 ppm, with the exception of Streptococcus mutans, Streptococcus gordonii and Streptococcus faecalis (all of which had MBC values of 10 ppm). The data show that 10 ppm silver binding in this invention exhibits the same or broader spectrum of activity than any other antibiotic tested. Antibiotics generally have a limited spectrum of antibacterial activity that affects only sensitive organisms, but according to the data, the silver composite of the present invention is equally effective against both gram positive and gram negative organisms. According to the low toxicity of silver preparations in general and the broad antimicrobial activity of this silver composite, this preparation can be effectively used as an alternative to antibiotics.

[238] D. Review of Previous Example [239] 1. US EPA IRIS Report on Silver-CASRN 7440-22-4 [240] 2. Fox CL, Modak SM. Effect of silver sulfadiazine on burn wound. Infections. Antimicrob Agents Chemother. 5: 582-588, 1974.

[241] 3. Furcher, JE, Richmond CR and GA Drake. Comparative radionuclide metabolism in mammals. IV. Maintenance of silver-llOm in mice, rats, monkeys and dogs. Health Phys. 15: 505-514,1968.

[242] 4. Grier, N. Silver and its components for disinfection, sterilization and preservation.

[244] Proof of the effect of 32 PPM silver composite on PSEUDOMONAS AERUGINOSA, SALMONELLA CHOLERAESUIS, and STAPHYLOCOCCUS AUREUS [245] A. Methods [246] Pseudomonas aeruginosa ATTCC # 15422, Salmonella choleraesuis ATTC # 10708 were tested on Staphylococcus aureus # 10708 Association of Official Analytical Chemists AOC Methods, Vol 1, 15th Edition, 1990 Arlington, VA) Official Methods 955.14, 95515 and 964.02. Nutrient Decant (NBAOAC) tubes were inoculated from the main culture and tubes were incubated at 37 + -2 ° C. Transfusion to clean feed tubes was performed over the following three days, with the final pass being maintained at 37 + -2 ° C.48-54 h. Pseudomonas culture was decanted into a fresh tube to remove the film. Other cultures were shaken for 304 seconds and then 10 min. at room temperature. Finally, the cultures were diluted 1: 100 in peptone water (PEPW) to which horse serum was added, resulting in a 5% organic presence. Test media (10 mm long polished 304 stainless steel cylinders with external diameter 8 mm and internal diameter 6 mm) were soaked in test solution for 15 min, removed, dried and dried at 37 ± -2 ° C for 40 ± min before use.

[247] Resistance to phenol. Five portions of one ml of each phenol solution without test were placed in a test tube and bathed in a 20 ± 2 ° C water bath. At 30-second intervals, 0.5 ml of each culture was added to the appropriate phenol solutions, shaken and returned to the water bath: After an appropriate exposure time of 5, 10 and 15 minutes, part of the suspension was poured from the test tubes and into the bovine broth tubes (LETH). . Tubes containing LETH were maintained at 37 + -2 ° C for 2 days.

[248] Carrier Titration. For carrier titration, only 10 ml peptide solution "Tween" (polysorbate type) (PEPT) was prepared. Two carriers were placed in individual tubes and a 1:10 solution was administered. The tubes were shaken vigorously to mix the bacteria in the solution. Different compositions were prepared and administered in 9 mL of LETH medium. The solution tubes were left for incubation at 37 ± 2 ° C. The last tube indicated the development of the logo organism on the carrier. The AOAC requires carriers to have a population of at least 1 χ 10 ⁴ cfu per carrier.

[259] A. Purpose of Example [260] The purpose of this example is to demonstrate the antibacterial activity of a silver-based variant of the present invention on a cow's side inoculated with a mixture of five strains of Salmonella or Escherichia coli 0157: h7 on a high inoculum solution. level (lx10 ⁶ cfu / cm ² ) and separately at low inoculum material solution level (lx10 ⁴ cfu / cm ² ) (cfu - colony forming unit).

[261] B. Material and Methods [262] Cow samples. Cow tissue samples were obtained from slaughterhouses within 8 hours after evisceration. The abdominal rectus muscle was detached from the carcasses that were hanging in the refrigerator, making an incision between the 11th and 12th ribs and then tearing the muscle along the natural seam. Aseptically obtained samples were placed in plastic bags on ice and transported with Multi-Vac (A-300) in an ice box at 4 ° C. Samples used for testing had a pH of 5.8 to 6.0 and had not been more than 36 hours after inoculation. 13 138 cm large samples were excised and processed from randomly selected abdominal rectus muscles. After treatment, a 3.5 cm ² flame sterilized stainless steel device and a surgical scalpel were used to aseptically obtain two meat samples at each sampling interval from each sample. Tissue samples were placed in a sterile bag containing 25 ml of 0.1% peptone and bagged for two hours (Lab Bender 400). Batch dilution was prepared and spiral muscle supernatants prepared at 0 minutes, 20 minutes, 1 hour, 4 hours and '24 hours after treatment in selected and restored medium.

[263] Bacterial cultures. Bacterial cultures were obtained from the Kansas State University (KSU) Starting Culture Collection and stored using the Protected Pellet Storage System. The following cultures were used for the Salmonella sample: S. lille (UGA), S. montevideo (UGA), S. typhimurium (UGA), S. agon (KSU 05 from CDC outbreak isolate) and S. newport (KSU 06 from CDC out of CDC outbreak isolate). The following cultures were used for the Escherichia coli sample: E. coli 0157.7 (CDC 01.03), E. coli 0157: H7 (USDA-FSIS 011-82 resistant 100 ppm), E. coli 0157: H7 (ATCC 43895) HUS associated with type I and II resistant toxins) and E. coli ATCC # 23740 (Genotype K-12 prototropic lambda).

[264] Stock cultures were cultured by placing one impregnated pellet in 5 ml Difco® tripty soy broth (TSB) solution and incubating for 24 hours at 35 ° C. The 0.05 ml loop from the appropriate culture was then inoculated in 5 ml of TSB solution and incubated for 24 hours at 35 ° C to obtain a pure culture. After incubation, 1 ml of the appropriate culture was inoculated with 49 ml of TSB and incubated for 24 hours at 35 ° C. After incubation, the samples were centrifuged (15,300 * g at 4 ° C) and the supernatant was decanted, but the pellet was resuspended in 50 ml of 0.1% peptone and re-centrifuged (15,300 * g at 4 ° C). C). The peptone was decanted and the remaining pellet was resuspended in 10 ml of 0.1% peptone. Five 10 ml culture vials were mixed together to form 50 ml compositions containing 10 ⁹ cfu / ml Salmonella genus. The mixture was diluted to 10 ⁶ cfu / ml or 104 ⁴ cfu / ml using 0.1% peptone. Cultures were confirmed by culturing on a selective and differential medium and by biochemical analysis of potential colonies using API 20E kits.

[265] Inoculation method. The cow flank specimen (rectum abdominal muscle) was cut to 13 χ 8 cm (104 cm ² ) and inoculated with a five-strain composition of Salmonella or Escherichia coli 0157: h7 at high inoculum material solution (10 ⁶ cfu / cm ² ). and alone at low inoculum material solution level (10 ⁴ cfu / cm ² ). This inoculum was sprayed onto the tissue surface using a plastic spray bottle with specimens contained in a sealed inoculum chamber. Actual concentrations of Salmonella on the surface of the meat were approximately 5.0 and 3.4 log cfu / cm ^{2 for} high and low level inoculum, respectively. For E. coli 0157: H7, the corresponding surface inoculations of the meat were 4.2 and 3.9 log cfu / cm ² .

[266] Cow specimens were then vertically suspended on stainless steel hooks attached to a motorized chain that transported cow specimens through a spray casing model (Kansas State University, Food Safety Laboratory) while being subjected to spray treatment. Treatments were made with both the silver composition of this invention and deionized water for a cow at 20 psi (pounds per square inch) for 20 seconds from a 13 cm wash under pressure in the body model. The spray nozzle (ΒΕΤΕ NF0580 303) provided approximately 20 ml of the solution on the surface of the cow sample. The solution and treatment application room temperature was approximately 14 ° C. After treatment, duplicate 3.5 cm ² samples were removed randomly from the side surface of the cow sample at 0, 20, 60 and 240 minutes. Samples were cultured and counted on a selective differential and restored surface. Logarithmic reductions were calculated by subtracting logio cfu / cm ² from inoculated / treated samples at the specified sampling times (0, 20, 60 and 240 minutes) from log 10 cfu / cm ² from uninoculated / untreated samples at 0 minutes. Sample processing included the use of 32 ppm silver, 22 ppm silver and 10 ppm silver compositions according to the present invention. Separately, combinations of 22 ppm Ag with 1.5% (w / w) hydrogen peroxide and 10 ppm Ag with 10 ppm peroxydisulfate (K ₂ S ₂ O 8) were tested.

[267] C. Results of Using a 32 ppm Silver Compound [268] The use of the invented 32 ppm compound resulted in a reduction in the bacterial count per piece of beef. Consequently, this reaction is expressed as logio of the number of bacteria in the control time 0 to the number of bacteria in the treated sample at the time of removal (i.e., the procedure).

[269] At the lowest baseline bacterial level of Salmonella (10 ⁴ ), the following logarithmic reductions were observed: 0.78 -0 minutes, 1.11-20 minutes, 1.08-60 minutes, and 1.23-240 minutes. Thus, 4 hours (240 minutes) the initial number of bacteria The bacteria in a control sample treated with 32 ppm silver is 10 ^{1 '23} .10 higher baseline For bacterial level (10 ⁶⁾ were found following logarithmic reduction: from 0.86 to 0 minutes , 0.95-20 minutes, 0.98 LV 13745 minutes, and 1.17-240 minutes. The results confirm that the inventive use of 32 ppm silver shows effective bactericidal activity against Salmonella in beef.

In addition, it should be noted that the disinfection of the meat surface is a major challenge for any disinfectant.

[270] At the lowest initial bacterial level of E. coli (10 ⁴ ), the following logarithmic reductions were observed: 1.03 -0 minutes, 1.28-20 minutes, 1.42-60 minutes, and 1.58-240 minutes. For higher baseline bacterial levels (10 ⁶ ), the following logarithmic reductions were observed: 0.65-0 minutes, 0.60-20 minutes, 0.83-60 minutes, and 0.87-240 minutes. The results confirm that the inventive use of 32 ppm silver 20 exhibits effective bactericidal activity against E. coli in the beef end.

[271] Results of 22 ppm silver compound application [272] Results of silver application in water. At the lowest baseline bacterial level of Salmonella (10 ⁴ ), the following logarithmic reductions were observed: 0.41

- 0 minutes, 0.43 - 20 minutes, 0.48 - 60 minutes, and 0.68 - 240 minutes. For the higher 25 baseline bacterial levels (10 ⁶ ), the following logarithmic reductions were observed: 0.24-0 minutes, 0.24-20 minutes, 0.42-60 minutes, and 0.61-240 minutes. The results confirm that the inventive use of 22 ppm silver provides effective bactericidal action against Salmonella in beef.

[273] Results from the application of silver in water and 1.5 wght% hydrogen peroxide.

At the lowest baseline bacterial level of Salmonella (10 ⁴ ), the following logarithmic reductions were observed: 0.34-0 minutes, 0.33-20 minutes, 0.36-60 minutes, and 0.62 30-240 minutes. For higher baseline bacterial levels (10 ⁶ ), the following logarithmic reductions were observed: 0.28-0 minutes, 0.14-20 minutes, 0.30-60 minutes, and 0.69-240 minutes. The results confirm that the invented 22 ppm silver total

The use of 1.5 wght% hydrogen peroxides provides effective bactericidal activity against Salmonella in beef [274] 10 ppm silver compound utilization results [275] Silver compound water utilization results. At the lower baseline Salmonella bacterial level (IO ⁴ ), the following logarithmic reductions were observed: 0.38-0 minutes, 0.41-20 minutes, 0.39-60 minutes, and 0.61-240 minutes. For higher baseline bacterial levels (10 ⁶ ), the following logarithmic reductions were observed: 0.24-0 minutes, 0.21-20 minutes, 0.41-60 minutes, and 0.54-240 minutes. The results confirm that the inventive use of 22 ppm silver provides 10 effective bactericidal activity against Salmonella in beef.

[276] Results of the use of a silver compound in water totaling 10 ppm KjS ^ CL,

At the lower baseline Salmonella bacterial level (IO ⁴ ), the following logarithmic reductions were observed: 0.26 to 0 minutes, 0.28 to 20 minutes, 0.35 to 60 minutes, and 0.58 to 240 minutes. For higher baseline bacterial levels (10 ⁶ ), the following logarithmic reductions were observed: 0.03-0 minutes, 0.16-20 minutes, 0.21-60 minutes, and 0.36-240 minutes. The results confirm that the use of 10 ppm silver in total 10 ppm potassium persulfate (K2S2O6) provides effective bactericidal activity against Salmonella in beef. 15 [277] EVIDENCE OF THE EFFECTIVENESS OF 10 PPM SILVER IN HUMAN DISEASE [278] Purpose of Example [279] The purpose of this example is to demonstrate the potential of the inventive silver-based compounds in the treatment of various human diseases. Research in this unit was conducted at the Air Force Base Hospital in Ghana, West Africa. Under the leadership of Quabiah, Korie-Bu

At the teaching hospital, Sr. Under the leadership of Sakey, and at the Justaba Clinical / Obstetric Hospital, Dr. Under the leadership of Abraham. A total of fifty-eight (58) patients were cured using the invented silver / water compound with 10 ppm silver. This compound was used both orally and externally as an alternative to traditional antibiotics. Diseases such as malaria, upper respiratory tract infections, urinary tract infections, sinusitis, vaginal yeast infections, eye, nose and ear infections, cut wounds, skin and fungal infections such as gonorrhea. 25 [280] B. Treatment Methods and Results [281] Abdominal Pain and Diarrhea. The method consists of approx. Drink 5-25 ml of the silver composition 1-5 times daily until the reaction is reported. One patient was treated with ca. 10 ml (about 2 teaspoons) of the composition described 3 times a day. The patient fully recovered within one day.

[282] Bronchitis. The method consists of approx. Drink 2 to 25 ml of the silver composition 1 to 5 times daily until the reaction is complete. Two patients were treated with ca. 5 ml (approx. 1 teaspoon) twice daily for 3 days. Patients were completely cured within three days.

[283] Vaginal Yeast Inflammation (Candida). The method consists of vaginal lavage with 5-25 ml of silver composition 1-5 times daily until reaction is reported. Five patients were treated with ca. 10 ml (about 2 teaspoons) of the composition described twice a day. Patients recovered fully within 6 days.

[284] Conjunctivitis. The method consists of dripping the silver composition into the infected eye 15 times a day until the reaction is reported. Two patients were treated with a drip of the infected eye twice daily. Patients recovered completely within one day.

[285] External wounds and infection (including Staphylococcus skin infections, septic ulcers and infected tumors). The method involves applying a silver composition to the infected body 1-5 times daily until a reaction is reported. Six patients were treated with ca. 5 ml (about one teaspoon) of the composition described twice a day. Patients recovered fully within three days.

[286] External otitis. The method consists of applying a silver composition to the infected ear 1-5 times daily until the reaction is reported. Six patients were treated with ca. 2 drops described compositions 3 times a day. Patients recovered fully within 4 days.

[287] Otitis Media. The method consists of applying a silver composition to the infected ear 1-5 times daily until the reaction is reported. One patient was treated with ca. 2 drops described compositions 3 times a day. The patient recovered fully within 4 days.

[288] Skin fungus. The method consists of topically applying a silver composition to the infected body 1-5 times daily until the reaction is reported. Two patients were treated with ca. 10 ml (about 2 teaspoons) of the composition described 3 times a day. Patients recovered fully within 8 days.

[289] Gonorrhea. The method involves applying a silver composition to the infected body until a reaction is performed. Two patients were treated with ca. 10 ml (about 2 teaspoons) of the composition described 3 times a day. Symptoms resolved within 6 days.

[290] Malaria. The method consists of drinking the silver composition 1-5 times daily until the reaction is reported. In the first study, 11 patients were treated with ca. 10 ml (2 teaspoons) of the composition described 3 times a day. Symptoms resolved within 5 days. Malaria protocols will be discussed in more detail below.

[291] Mouth odor and gum inflammation. The method consists of mouth rinsing with a silver composition 1-5 times daily until a reaction is reported. 2 patients were treated with a mouthwash composition. Symptoms resolved completely within 3 days (inflammation of the gums) and 1 day (odor of the mouth).

[292] Pelvic inflammation. The method consists of vaginal lavage with 5-25 ml of silver composition 1-5 times daily until reaction is reported. One patient was treated with ca. 5 ml (about 1 teaspoon) of the composition described twice a day. The patient's symptoms resolved within 5 days, [293] Pharyngitis. The method consists of rinsing the neck with a silver composition 1-5 times a day until the reaction is reported. 4 patients were treated with ca. 10 ml (2 teaspoons) of the composition described 3 times a day. Patients recovered fully within 6 days.

[294] Retroviral infection (CIV). The method consists of drinking 5 to 40 ppm (parts per million) of the silver composition 1 to 5 times daily until the reaction is reported. One patient with human immunodeficiency virus was treated with ca. 5 ml (about 1 teaspoon) of the composition described twice a day. The patient's symptoms completely resolved within 5 days.

[295] Sinusitis and rhinitis. The method consists of applying a silver composition to the nose 15 times daily until the reaction is reported. Six patients with nasal infections (four with sinusitis and two with rhinitis) were treated with ca. 2 drops of the composition described in the nasal passages 3 times a day. Patients recovered fully within 4 days.

[296] Inflammation of almonds. The method consists of rinsing the neck with a silver composition 15 times daily until the reaction is reported. One patient was treated with the described composition 3 times daily. The patient recovered fully within 7 days.

[297] Upper respiratory tract infection. The method consists of drinking the silver composition 1-5 times daily until the reaction is reported. 2 patients were treated with ca. 5 ml (about 1 teaspoon) of the composition described 3 times a day. Patients recovered fully within 6 days.

[298] Urinary tract infections. The method consists of drinking the silver composition 1-5 times daily until the reaction is reported. 3 patients were treated with ca. 10 ml (2 teaspoons) of the composition described 2 or 3 times a day. Patients recovered fully within 6 days.

[299] C. Discussion [300] These results are consistent with the various in vitro assays reported herein. The silver compound is extremely effective in combating a large number of microbes in vitro. However, tests show that this efficacy is not lost even in the presence of large amounts of organic material. Silver compositions are effective in vivo, where the organic background is very high. Many other disinfectants are ineffective in the presence of large amounts of organic material and are too corrosive or toxic for in vivo use.

[301] Complementary studies of malaria in Ghana, Africa.

[302] Another more regulated study was conducted in Ghana. The aim of the study was to apply a very specific protocol and to focus solely on the healing properties of the 10 ppm silver / water composition described herein for malaria patients.

[303] The purpose of the protocol was to develop a procedure whereby a 10 ppm silver / water composition produced by the methods could be screened for its potential curative properties in the treatment of patients infected with one of the 4 species of Plasmodium. The following is an overview of the protocol:

[304] Studies were conducted by medical doctors (MDs) experienced in treating the disease in medical clinics or hospitals. A total of 16 patients treated with the silver formulation twice daily for 5 days were subjected to blood tests one day prior to the start of the study, and then daily until blood tests showed neutralization of the parasite for at least 2 days. Patients were only paid if they fully adhered to the silver composition and blood test schedule.

Composition of the protocol:

Number of doctors (MD): 2

Number of patients studied: 16-8 males and 8 females [305] Total study duration: 15 days [306] 10 ppm silver / water composition dose for patient treatment: total daily dose per ounce divided into two equal doses: one half-ounce (3 teaspoons) in the morning and one half-ounce (3 teaspoons) in the evening. Patients were treated with a silver / water composition for the first 5 days of the study, and if the parasite was not neutralized within the first 5 days, treatment was continued until the parasite was cleared or by day 15.

[307] In the case where the parasite had disappeared by the second or third day, the application of the silver / water composition was continued until day 5, and a record was made of when the parasite completely disappeared.

[308] In the case where the patient was still carrying the parasite after 15 days of treatment with the silver / water composition, the study was discontinued and a failure of the patient was recorded.

[309] A patient who recovered in less than 5 days was recorded the date of complete elimination of the parasite and continued to be treated with the silver / water composition until day 5 and enrolled in the study until day 15.

Blood tests:

[310] Blood tests used: The presence (or absence) of a parasite in a patient's blood was determined by the Acridine Orange staining test or the Giemsa staining test by examining thin or thick blood stains from each patient. The patient's blood was tested on day zero to determine that they did indeed have active malaria. If the blood test confirmed the presence of active malaria, the patient was examined for acceptance in the study. The examination included recording the necessary data, namely, name, age, onset of disease, the patient was informed of what was expected of him during the study and what amount he would be paid for full compliance, and he was informed that non-compliance would lead to Withdrawal from the study, without payment, i Patients who agreed to participate in the study were given instructions describing the use of the silver composition, blood tests and emphasizing the need for full compliance.

The protocol was fully respected during a recent study in Ghana, Africa. All patients received the same dose and their blood was analyzed daily for the presence of plasmodia parasites. Table 4 presents parts of the previous studies (Study 1 and Study 2) as well as the new Study 3, which followed the above protocol.

[311] Summary

Table 4

Object of research 1 2 3 4 Number of patients 11th 16th 16th 13th Age group 8-75 2-90 3-61 15-57 Men / Women No data No data 8/8 6/7 Average daily dose 10 ml 5 ml 15 ml 15 ml shortest recovery time + 3 days 3 days 2 days 3 days The longest recovery time 7 days 10 days 8 days 6 days average recovery time 5 days 6.3 days 4.3 days 4 days # patients tested for plasmodia 0 7th 16th 13th # patients with plasmodia 0 0 0 * 0 * # Treatment errors 0 0 0 0

+ Recovery time was based on patients who, according to doctors, had no symptoms.

* Each of these patients was tested daily for 14 days. After six days, each of the blood samples taken showed a negative plasmodia test result.

[312] The 10 ppm silver / water solution of the present invention has obvious positive results in the fight against malaria-causing parasites.

[313] EVIDENCE OF 100 PPM SILVER EFFICIENCY

IN VITRO TREATMENT OF MALARIA (314) INTRODUCTION 15 [315] Globally, malaria has been and continues to be one of the most important public health threats. The cause of the disease is at

A parasitic protozoan of the genus Plazmodium. This organism has a complex life cycle, during which the parasite changes through sexual reproduction in the invertebrate (ode) host and subsequent asexual reproduction in the vertebrate host, and in addition to malaria, birds and reptiles can be the host of malaria. One part of the life cycle of the mosquito is the sporogonal phase, which culminates in the formation of sporozoites, which are fed to the vertebrate host during feeding by the vector.

Sporozoites develop to the fission phase, with a rapid proliferation of erythrocytic and non-erythrocytic sites. The parasite is extracellular during its sporogonal phase, becoming intracellular at the stage of division of its development. Laboratory cultivation of parasites requires equating conditions in the sporogonal phase of the life cycle with those of the vector of infectious disease, but for the fission phase, to create conditions conducive to growth of erythrocytic and non-erythrocytic sites in vertebrate hosts.

[316] Malaria is one of the most common parasitic diseases in the world, ranked at least third in the world among major infectious diseases in terms of mortality. The disease is caused by a parasitic protozoan of the genus Plazmodium. There are the following malaria-causing plasmodia species: Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae and Plasmodium ovale. Vectors are mainly malaria mosquitoes, although malaria infection can also be obtained from contact with infected blood, such as by blood transfusion.

[317] Classic symptoms of malaria include fever, headache, chills, vomiting, trembling and convulsions. With some rare forms of Wednesday malaria, fever and chills may not be present, but the patient may have nightmares or coma.

Remission periods can last from a few weeks to several months. Often, the cause of death from malaria infection is severe anemia.

[318] Plasmodium falciparum:

This parasite has some essential features. These include the crescent-shaped gametocyte, its slow growth rate, and its pigmentation around the nucleus (perinuclear arrangement), which is not observed in the gametocytes of other primate malaria parasites.

P.falciparum differs from other human parasite species with higher virulence and lethal effects, although the fission and erythrocytic stages are largely confined to the internal organ capillaries and sinusoidal capillaries. A popular name for this 30 disease caused by P.falciparum is "Malignant Trinitarian Malaria".

MATERIALS AND METHODS

Citrate salt water:

Sodium chloride 9gm

Sodium citrate 20 gm

Distilled water 1000 ml.

Gimsa coloring:

Gimza satin powder 75 gm

Pure alcohol 75 ml

Glycerin ml

Field Color:

Field Solution # 1

1. Dissolve 1.6 g of methylene blue dye in 1 liter of distilled water.

2. Dissolve 2.6 g of Na ₂ HPO ₄ (anhydrous) in the solution obtained in 1 step.

3. Dissolve 1 g of openwork solution 1 in Step 2.

4. Dissolve 2.6 g of KH ₂ PO ₄ in Step 3.

5. Place on lightly warm surface, stir or shake for 45 minutes to 1 hour.

6. Store the solution at room temperature for 24 hours.

7. Filter

Field Solution # 2

1. Dissolve 2 g. Eosin Υ 1 liter distilled water.

2. Dissolve 2.6 g of Na ₂ HPO _{4 in the} resulting solution.

3. Dissolve 2.6 g of KH ₂ PO ₄ in Step 2.

4. Filter

Wright coloring

White powder 6.0 g

Colored powder 0.6 g

Methanol 1000 ml

Stir overnight and filter before use.

Human serum / AB + blood group plasma

Human serum / plasma A + blood

RPMI- 1640 Incomplete Environment (Personal Contact with Dr. Sutaru, Haffkine Institute, Parela)

RPMI - 1640 Complete Environment (Personal Contact with Dr. Sutaru, Haffkine Institute, Parela)

COLLECTION AND TREATMENT OF INFECTED BLOOD [319] Parasite-infected erythrocytes were obtained by venipuncture collection of 6 ml aliquots of blood in 1 ml citrate saline from patients at the Hospital for the Study of Infectious Diseases of the Castle, Bombay, and Plodium v. Blood samples were placed in sterile 10 ml vials. Samples were studied by preparing smears and staining with 10% Gimza dye / Field dye / Rye dye to detect and confirm the presence of malaria-causing parasites. The level or percentage of sample parasitemia was recorded in writing.

[320] Blood cells affected by the parasite were washed twice with incomplete preparation and once with complete preparation and a 6% cell suspension in complete preparation was prepared. Cultures were prepared by placing 0.5 ml of suspension in each Petri dish. 1.5 ml of complete preparation was added to it and the dishes were stored in an atmosphere of 5% CO2 and 4-17% O2. The preparation was changed daily by suctioning the old preparation with a sterile pipette and adding 1.5 ml of complete preparation. Cultures were maintained by adding fresh cells after one week (from blood groups A + or AB +; rinsed and similarly prepared cell suspension), washing twice a week until the expected parasitic index reached> 1%. If the baseline parasitic index was greater than 1%, the Blood Nutrient Mixture (BMM) was immediately used to determine sensitivity to the drug. (Thanh, 2001) and (Tasanor, 2002)

SCRAP PREPARATION AND STAINING PROCEDURE [321] Cultures were washed twice a week. For washing, cultures were removed from the dishes and placed in centrifuge tubes. About 5 ml of incomplete preparation was placed in each centrifuge tube and mixed thoroughly. The tubes were centrifuged for approximately 10 minutes at 1000-1500 rpm. After about 10 minutes, the tubes were removed from the centrifuge and the supernatant fluid was removed. The cultures were then subjected to two similar washes, one with incomplete RPMI-1640 and the other with complete RPMI-1640. After three washes, the cultures were transferred to separate Petri dishes. A swab was prepared from each culture and stained with Gimza / Field / Rita dye. Approximately 1.5 ml of preparation was added to each dish and the swabs were examined under an optical microscope to determine the parasitic index and whether% parasitemia was obtained for each culture. Fresh red blood cells were added to each dish weekly (Pradhan, 1984).

[322] PREPARATION OF STRAINS [323] A drop of culture was taken from a dish on a microscope slide. A plan smear was performed, which was air dried. The smear was fixed by immersing the slide in a connected container of pure alcohol. A 10% Gimza dye solution was prepared and used to stain the swabs. The slides were kept immersed in 10% Gimza dye solution for approximately 30-40 minutes and then washed under tap water.

[324] Parasite Presence Index [325] Parasite Presence Index was calculated by counting the number of parasites per 100 erythrocytes in blood smears. At least 100 types or 10,000 RBCs were screened for this purpose.

[326] Cultural systems

Culture dishes were prepared with 5% hematocrit and approximately 1% parasitemia. The lower the initial parasitemia, the greater the increase in parasite numbers in in-vitro cultivation.

[327] SENSITIVITY TO MEDICINAL PRODUCTS [328] 16 mm flat bottomed Microwell dishes were used to test for sensitivity to therapeutic agents. One dish was used per sample. The first two dishes were used for drug sensitivity testing and received 50µ1 patient BMM or culture and 50µ1 RPMI complete preparation and no drug. For testing, 50μ1 of culture or patient BMMs were mixed in a container containing 50μ1 of special silver nanoparticle (ESNP) at different concentrations. Microwell i plates were coated and heated in a 37 ° C heater for approximately 49 hours.

Most of the parasites entered the division phase after a 48 hour incubation period. After incubation with a micropipette, the supernatant was removed; Blood was drawn from each vessel to prepare smears and examine for development of sharing. Test results were evaluated by counting the parasites on the stained preincubation and postincubation plates and the delays associated with ESNP in schizont formation. The test reliability in the control vessel must be> 10% maturity of the schizont. (Wernsdorfer and Wemsdorfer, 1995).

Results

Species % Reduction in parasites Plasmodium falciparum 94% Plasmodium vivax 92% Plasmodium berhei 90%

[329] An in vitro assay used to determine the efficacy of an antimalarial agent shows that the ESNP of -100 pm is capable of reducing parasite numbers in vitro. This is significant because the parasites were collected from patients with chills and elevated body temperature and other classic symptoms of malaria infection.

[330] EVIDENCE OF THE EFFECTIVENESS OF 10 PPM SILVER AGAINST TUBERCULOSIS BACTERIA [331] A. Purpose [332] The purpose of this example is to demonstrate the effectiveness of the present invention, a silver composition, in the fight against tuberculosis-causing bacteria. This example describes procedures for evaluating the effectiveness of the present invention in combating tuberculosis agents. This methodology is based on the activity test of the mycobacterium tuberculosis agent. The method was approved by EPA on December 11, 1985 (refer to United States Environmental Protection Agency, 1986. Office of Pesticides and Toxicants. Request information on efficacy data for mycobacterial tuberculosis eradication agent included in "All Antimicrobial Pesticides with Tuberculosis Mycobacterial Eradication Agent" description "(received 1986

June 13).

[333] B. Materials and Methods [334] Materials. The inventor of the invention - the silver composition consists of 10 ppm silver and water. The silver composition has been found to use a liquid matrix against Mycobacterium bovis BCG (TMC 1028). This organism causes tuberculosis in animals and can cause tuberculosis in humans. It is used as a "surrogate" for M. tuberculosis, a major cause of human tuberculosis, as tests show that it has a similar susceptibility to M. tuberculosis. The experimental organism was exposed to the silver composition twice 4 times and repeatedly using membrane filtration.

[335] Procedure. The frozen stock culture vial was removed from storage and thawed. An equal amount of buffer gelatin (BUGE) was added to the cell suspension and homogenized with a Teflon ® (polytetrafluoroethylene type) tissue homogenizer for 1 minute while maintaining the culture temperature in the ice bath at 0 to 4 °. The homogenized tissue suspension was diluted with saline Tween®80 (polysorbate type) (ST 80) to about 10 ⁷ cfu / ml.

[336] Control infection titration. Culture ten-fold serial dilutions were prepared in reconstituted media containing 9 mL of Neutralizing Liquid Medium (NEUB) at 10 ' ⁶ dilution. Three 1 mL aliquots of the respective dilution were filtered through a membrane filter, first adding 10-20 mL of saline (PHSS) in the filter and then adding 1 mL aliquot of the respective solution. The filter was then rinsed with approx

100 mL PHSS. Filters were removed from the filter casing and maintained on sterile 7H11 agar plates. The plates were heated in a humidified chamber for 21 days at 37 +/- 2 ° C.

[337] Positive control. A tube containing 9 ml of ST80 was prepared and the temperature equated to 20 +/- 0.5 ° C. During this time, 0.1 ml of test organism cultures was added to the contents of the tube (1:10 dilution). The sample was held for 60 minutes. Ten-fold serial dilutions of culture were prepared in dissolution blanks containing 9 ml NEUB) in 10 ' ⁶ dilution. Three 1 ml aliquots of the respective dilutions were filtered with a membrane filter, first adding 10-20 ml PHSS to the filter, then adding an ml aliquot of the respective solution. The filter was then rinsed with about 100 mL PHSS. Filters were removed from the filter casing and maintained on sterile 7H11 agar plates. The plates were heated in a humidified chamber for 21 days at 37 +/- 2 ° C.

[338] Test. Two 25 X 150 mm tubes containing 9 ml of test sample were cooled to 20 +/- 0.5 ° C in a water bath. To each tube containing the disinfectant to be tested (ie silver composition) was added 1 ml of the culture of the test organisms. The tube was mixed by cutting and placed back in the water bath. At 15, 30, 45, and 60 minutes, 1.0 mL aliquots of cell suspension of the disinfectant were transferred to 9 mL of NEUB and mixed thoroughly. Ten-fold serial dilutions of culture were prepared in dissolution blanks containing 9 ml NEUB 10 ' ⁶ dilution. Three 1 ml aliquots of the respective dilutions were filtered with a membrane filter, first adding 1020 ml PHSS filter, then adding 1 ml aliquot of the respective solution. The filter was then rinsed with about 100 mL PHSS. Filters were removed from the filter casing and maintained on sterile 7H11 agar plates. The plates were heated in a humidified chamber for 21 days at 37 +/- 2 ° C.

[339] Phenol control. The culture was tested in 0.8% phenol solution to demonstrate minimal culture viability and resistance. A 1 ml aliquot of the culture of the test organism was placed in 9 ml of phenol solution equilibrated at 25 +/- 0.5 ° C and heated for 20 minutes. After the exposure period, 1 mL of phenol / body solution was removed and 9 mL of NEUB was added. Ten-fold serial dilutions of culture were prepared in dissolution blanks containing 9 ml NEUB 10 ' ⁶ dilution. Three 1 mL aliquots of the respective dilutions were filtered with a membrane filter, first by adding 1 ΟΣΟ mL to the PHSS filter, then adding an Inti aliquot of the respective solution. The filter was then rinsed with about 100 mL PHSS. Filters were removed from the filter casing and maintained on sterile 7H11 agar plates. The plates were heated in a humidified chamber for 21 days at 37 +/- 2 ° C.

[340] Neutralization verification. To the 1 ml aliquot of the disinfectant was added 8 ml NEUB. The temperature of the disinfectant / neutralizer liquid medium was set to the same temperature as the test samples. One ml of test organism cultures was added to the mixture and mixed thoroughly. Incubation was continued for a period of time necessary to filter the sample. An additional 9 ml NEUB was added to 1 ml aliquot of test organism and mixed well (1:10 dilution). Ten-fold serial dilutions of culture were prepared in dissolution blanks containing 9 ml NEUB) in 10 ' ⁶ dilution. Three 1 ml aliquots of the respective dilutions were filtered with a membrane filter, first adding 10-20 ml PHSS filter, then adding 1 ml aliquots of the respective solution. The filter was then rinsed with about 100 mL PHSS. Filters were removed from the filter casing and maintained on sterile 7H11 agar plates. The plates were heated in a humidified chamber for 21 days at 37 +/- 2 ° C.

[341] C. Results [342] The initial titre of the culture to be tested was 4.7 X 10 ⁷ cfu / ml. The positive control titer was 6.5 X 10 ⁶ cfu / ml. The techniques used in this study effectively demonstrated neutralization with 95.2% recovery in the disinfectant / neutralizer solution as compared to the mass challenge.

[343] The predicted numerical values for the test plates were underestimated and therefore the reported numerical values indicate ">", noting that the predicted numerical value and the well-defined numerical value for the plate solutions are outside the limits of detection.

[344] When calculating the logarithmic and percentage reductions of the disinfectant relative to M.bovis, the predicted calculations were higher, but only resulted in "lower" logarithmic and percentage reductions ('<'). The purpose of the calculation is to demonstrate that the predicted numerical value and the well-defined numerical value for plate solutions are outside the limits of detection. All reductions were calculated using the positive control as the initial start titre for the organism. The results of logarithmic and interest reduction are summarized in the table below. As shown by the culture resistance measurements under study, M.bovis phenol resistance exhibits a ~ 1.81 logarithmic reduction at 20 minutes exposure to 0.8% phenol.

Logarithmic reduction Percentage <0.12 <12.3% <0.22 <40.0% <1.57 <97.2% <1.56 <97.2%

Logarithmic reduction Percentage <0.26 <44.8% <0.20 <36.9% <1.58 <97.3% <1.53 <97.1%

[345] First iteration:

Exposure time reduction minutes 30 minutes 45 minutes 60 minutes [346] Second iteration

Exposure Time Reduction Minutes 30 Minutes 45 Minutes 60 Minutes [347] D. Conclusions [348] The use of the inventive silver compound is effective against tuberculosis bacteria. A method that incorporates the methods of applying this inventive silver compound is effective in combating tuberculosis organisms.

[349] EVIDENCE OF THE EFFECTIVENESS OF 10 PPM SILVER IN THE FIGHT AGAINST CANDIDA ALBICANS AUC # 10231, TRICHOMONAS VAGINALIS NTCC # 2W235 UNMRSA STAPHYLOCCOCUSAUREUS ATCC # BAA-44 [3501] The purpose of this example is to exemplify [351] Candida albicans ATCC, Trichomonas vaginalis ATCC 20235, and Staphylococcus aereus ATCC BAA-44.

[352] Candida albicans, yeast sheaths, and Trichomonas vaginalis, protozoa, can cause many health problems, including vaginal infections, rashes, and thrush. The results below show that the invented silver compound is capable of killing almost 100% of both types of organisms. The results confirm the applicability of the inventive silver compounds in rash and feminine hygiene products.

[353] Staphylococcus aereus, if it occurs in the wound, can cause serious blood poisoning. It was once treated without problems with penicillin, but the body is currently mutated and is completely resistant to penicillin. The next antibiotic to be used is methicillin, but methicillin-resistant strains are becoming more common, especially in hospitals. These strains are known as MRSA (methicillin-resistant Staphylococcus aereus), which is unofficially referred to as a "super insect" (suberbug). People with MRSA infection can die within a few days. The results of this study confirm that the invented silver compound destroys 91.6% of MRSA in just 10 minutes and 99.5% in an hour. The results confirm the effectiveness of the invented silver compound in destroying MRSA, which is known as a dangerous infection.

[354] B. Methods and Results [355] Using the USP Defense Rapid Test Assay in conjunction with the present invention, which incorporates 10 ppm silver in water, results have been obtained which demonstrate that the inventive silver compound can be so effectively used against yeast infections, protozoal infections and treatment-resistant bacterial infections.

[356] Candida Albicans ATCC # 10231. Initial Candida Albicans yeast concentration was 6.8x10 ⁵ cfu / ml. No colonies were detected after contact with the silver compound for either 10 minutes, 30 minutes, 1 hour, or one day.

[357] Trichomonas vaginalis ATCC 20235. Initial concentrations of Trichomonas vagmalisvi & aspi were 6.8x10 ⁵ cfu / ml. After contact with the silver compound for either 10 minutes, 30 minutes, 1 hour or one day, 0% motility per 100 organisms was observed. This means that one hundred (100) parasites of Trichomonas vaginalis were examined under a microscope to determine the motility of the pups. None of the one hundred (100) t parasites showed motility until ten (10) minutes after contact with the silver compound, which demonstrates the suppression and destruction properties of this silver compound parasite. Twenty-five (25) percent of the outer membranes of the parasites were ruptured after one (1) day of contact.

[358] Staphylococcus aereus ATCC BAA-44 Initial methicillin-resistant Staphylococcus aereus (MRSA) concentration was 6.0x10 ⁵ cfu / ml. After contact with said silver compound, after 10 minutes contact 500,000 cfu / ml (91.6% destroyed), after 30 minutes contact 70,000 cfu / ml (98.8% destroyed), after 1 hour contact 30.00 cfu / ml (99.5% destroyed), and less than 10 cfu / ml after one day of contact (virtually completely destroyed).

[359] PPM SILVER, 14 PPM SILVER + 1.5% H ₂ O ₂ , AND 22 PPM SILVER CERTIFICATE OF EFFECTIVENESS AND LACK OF CYTOTOXICITY IN DNA POLYMERASE LOSS AND REVERVERSE TRANSCRETE A. [36] and [ The aim is to illustrate the effectiveness of the inventive silver compounds against hepatitis B. The example shows that the given silver compounds have antiviral activity. Any agent used in antiviral therapy required negligible or non-existent cytotoxicity, as silver compounds were analyzed for cytotoxicity [362] Hepatitis B is caused by a DNA virus belonging to the hepadnaviridae family. Hepatitis B virus (HBV) is a 3.2 kb DNA virus that mainly develops in liver cells (hepatocytes). Reproduction involves two major enzymes: DNA polymerase and reverse transcriptase. The results of this example confirm that the inventive silver compounds inhibit replication in either DNA polymerase or reverse transcriptase. The results of this example prove that the inventive silver compounds have antiviral activity. Likewise, the results of this example show that the inventive silver compounds can be effective in the fight against hepatitis B.

[363] Further information, when hepatitis B enters a new host, it infects the liver if the host's immune system is not strong enough. During the infection, the virus attaches to the membrane of the liver cell, releasing its own genome that enters the liver cell. The genome then inserts its DNA and DNA polymerase contents into the nucleus of the liver cell. In liver cells, virus replication occurs by reverse transcriptase and a transformation process involving the enzymes of reverse transcriptase and DNA polymerase. DNA polymerase causes the liver cell to make copies of the hepatitis B DNA. These copies of the virus enter the bloodstream through the liver cell membrane. From here they can infect other liver cells and replication is even more efficient. The incubation period for hepatitis B virus is approximately 6 to 25 weeks (i.e., before the appearance of generally identifiable histological and physical symptoms). However, there are several biochemical and histological changes that occur in the early stages of hepatitis B infection.

[364] B. Materials [365] Solutions containing 10 ppm, 14 ppm, 22 ppm and 32 ppm silver compounds according to this invention are used. The nucleotides dATP, dGTP, dCTP and [ ³ H] -dTTP were obtained from standard commercial sources, as well as pharmaceuticals such as lamivudine (a synthetic antiretroviral agent) and zidovudine (AZT). Isolated, fresh hepatitis B virus was obtained from a person suffering from hepatitis B infection, sampled by Haffine Institute, Mumbai INDIA (WH0 Certified Testing Laboratory). Test cell cultures (Wo and Hep2) were grown as monolayers using typical cell culture techniques.

[366] C. Methods [367] 1) Procedure for DNA Polymerase Inhibition Test [368] General approach. Hepatitis B virus extract from human body was kept in an incubator with nucleotides labeled with radioactive isotope and active inhibitor. Percent inhibition is calculated based on the amount of newly synthesized viral nucleic acid with lamivudine as a positive control and PBS as a negative control.

[369] Special procedure. The isolated hepatitis B virus was cleaved to an extraction-free polymerase enzyme free of contaminating enzymes. A viral extract containing nucleotides (25 mL) of dATP, dGTP, dCTP and [ ³ H] -dTTP was added to the reaction mixture. An active inhibitor was added to the mixture consisting of viral extract and nucleotides. The resulting mixture was kept in an incubator for 24 hours at 37 ° C.

[370] A separate negative control experiment was performed using phosphate buffer (PBS, 3ml) instead of inhibitor (3ml).

[371] A separate positive control experiment was performed using a known DNA polymerase inhibitor (3 mL lamivudine at 3 mg / mL) in place of the inhibitor tested (3 mL) [372] The reaction was stopped by the addition of 25 mL of EDTA and 25 mL of TCA ( trichloroacetic acid). The reaction mixture was then applied to ionic paper (DEAE paper). The paper was washed three times with TCA and then with ethyl alcohol. The filter paper was air dried and placed in a scintillation vial along with a scintillation cocktail (Blue Star). For count control, the blank silver composition assay was subjected to a complete procedure without virus loading to test for any potential interference with the scintillation count method.

 [373] This method is described in the paper: P.S. Venkateswaran, I.Millman and B.S. Blumberg, "Bffect of an extract from Phyllanthys niruri on hepatitis B and woodchuk hepatitis virus: in vitro and in vivo studies," Proc. Nati. Acad.Sci., USA, 84, 274-278 (1987), which is incorporated herein by reference.

[374] 2) Reverse Transcriptase Assay Procedure [375] Molecular murine leukemia virus reverse transcriptase (MoMuLV), which has commercially prepared virus enzymes for Poly (A) dt (basic RT), was used. 50 ml of MoMuLV preparation was combined with a mixture of dATP, dGTP, dCTP and [ ³ H] -dTTP nucleotides.

[376] This mixture was combined with 3 ml of the inhibitor to be tested and the resulting mixture was incubated for 24 hours at 37 ° C.

[377] A negative control experiment was performed using phosphate buffer (PBS, 3ml) instead of inhibitor (3ml).

[378] A positive control experiment was performed using a known reverse transcriptase inhibitor (3 ml AZT at a concentration of 0.625 micrograms / ml) as the inhibitor site (3ml).

[379] The reaction was stopped by the addition of 25 ml EDTA and 25 ml TCA. The reaction mixture was then applied to ionic paper (DEAE paper). The paper was washed three times with TCA and then with ethyl alcohol. The filter paper was air dried and placed in a scintillation vial along with a scintillation cocktail. Radioactivity was measured with a liquid scintillation counter (Blue Star).

[380] 3) Cytotoxicity Testing Procedure [381] Cells from healthy, confluent Vaud and Hep2 dog cultures were maintained and maintained through flow every 3-4 days. One day before the assay, cells were cleared from cultures using standard technology and suspended in nutrient medium and plated in microtiter plates and placed in a 5% CO ₂ incubator at 37 +/- ° C. An aliquot (100 mL) of each test substance was placed in a well (triplicate) with 100 mL PBS for control. The wells were examined under a microscope for 25 4 4 hours for cytopathic effect (CPE). All results are shown in Table 5 below.

[382] D. Results [383] Reverse transcriptase inhibition test results

Table 5a

Sample Inhibition%

Negative control (PBS) 0

Positive control (OCT)

Silver, 10 ppm 89.52

Silver, 14 ppm and 1.5% H2) 2 86.93

Silver, 22 ppm 84.46 [384] DNA polymerase inhibition test results

Table 5 b

Sample Inhibition%

Negative control (PBS) 0

Positive control (lamivudine)

Silver, 10 ppm 77.73

Silver, 14 ppm and 1.5% H2) 2 65.6

Silver, 22 ppm 60.89 [385] The inventive silver compounds demonstrate high efficiency in inhibiting DNA polymerase.

[386] Reverse transcriptase inhibition test results:

Table 5c

Sample Inhibition%

Negative control (PBS) 0

Positive Control (OCT) 18.06

Silver, 10 ppm 89.52

Silver, 14 ppm and 1.5% H2) 2 86.93

Silver, 22 ppm 84.46 [387] Thus, the inventive silver compounds inhibit reverse transcriptase. The inventive silver compounds can be considered effective against viral diseases such as hepatitis B.

[388] Results of the cytotoxicity test: Table 5d

Sample Control (PBS)

Silver, 10 ppm

Silver, 14 ppm or 1.5% H2) 2

Silver, 22 ppm

Watch for Hep2

No CPE No CPE

No CPE No CPE

CPE positive CPE positive No CPE No CPE [389] These results show. That silver compound is essentially non-cytotoxic. As expected, hydrogen peroxide, which is known to be cytotoxic, exhibits a cytotoxic effect. Thus, silver is harmless to cells when used in vivo.

[390] 12. PROOF OF EFFECTIVENESS OF SILVER COMPOUND AS A WATER DISINFECTANT [391] A. Purpose [392] Tests have been conducted to demonstrate the effectiveness of the inventive silver compound in disinfecting drinking water.

[393] B. Methods [394] Two doses of Klebsiella oxtyoca were added to a sample of untreated water. 100 ml aliquots of this aqueous solution were added to 0.05 ppm, 0.1 ppm, 0.2 ppm, 0.5 ppm or 1.0 ppm of the inventive silver composition. After an incubation period of 5-60 minutes, the sample policy was filtered through a membrane filter. The filter was rinsed with approximately 100 ml sterile water. The filters were sterile removed from the filter casing and placed on agar plates with intestinal rod medium. The plates were kept in the incubator for 24 hours, providing growth conditions and counting was performed.

[395] Table 6

Sample Silver (ppm) Contact (min) Total intestinal sticks (per ml) Cfu / 100 ml Untreated water - - 36 TNTC 1 1.00 5.0 0 0 2 1.00 10.0 0 0 3 1.00 15.0 0 0 4 1.00 30.0 0 0 5 0.50 10.0 0 0 6th 0.50 30.0 0 0 7th 0.50 60.0 0 0 8th 0.20 5.00 0 0 9th 0.20 10.0 0 0 10th 0.20 30.0 0 0 11th 0.20 60.0 0 0 12th 0.10 10.0 0 0 13th 0.05 20.0 0 0

TNTC = Too Much to Count (396) The silver compound tested proved to be surprisingly effective, even in the shortest time period (20 min) given to the lowest concentration test (0.05 ppm, all bacteria were killed. At 0.20 ppm and higher bacteria are completely eradicated within 5 minutes, so it is clear that complete eradication takes less than 5 minutes.

[397] PROOF OF EFFECTIVENESS OF PPM SILVER COMPOUND AS A SURFACE DISINFECTANT a product that can be used in hospitals, medical environments, residential homes, commercial premises and offices. It has been found to be suitable for use against the most dangerous pathogens, which include: Gram-positive bacteria such as Staphylococcus aureus (nowadays recognized as the most dangerous bacterium)

US hospitals), Gram-negative bacteria such as Salmonella choleraesuis (blamed for food poisoning) and hospital or hospital-acquired pathogens such as

Pseudomonas aeruginosa (often found in burns and wounds).

[399] The silver compounds of the present invention can be sprayed on and off work areas without endangering human or animal health or welfare. It can be used to disinfect surfaces such as walls, tables, chairs, lighting fixtures, bathrooms, glass, porcelain, metal, glazed ceramics, enameled or painted surfaces by spraying or by wiping with a cloth dampened with the inventive silver compound. . the selected disinfection method involves one or more steps to clean the surface to be disinfected using a spray gun, broom, sponge, or cloth provided with a silver compound, thoroughly moistening the area to be disinfected, allowing the surface to remain moist for at least 10 minutes. relationship can be determined.with the help of the Arrenius equation or other means known to the individual) and wiping the surface with a clean paper or cloth towel. Compounds for surface disinfection include 5 to 40 ppm silver compounds. The most suitable silver compound for surface disinfection of this invention comprises (32 +/- 3) ppm silver. Another silver compound of the present invention suitable for surface disinfection contains (10 +/- 2) ppm silver. Another silver compound of the present invention suitable for surface disinfection contains (22 +/- 2) ppm silver.

[400] PROOF OF EFFECTIVENESS OF SILVER COMPOUND AS SUPER DISINFECTANT [401] A. Purpose of Example [402] The purpose of this example is to demonstrate the antimicrobial activity of the silver compound of the present invention (given at 10 ppm silver, 14 ppm silver, 14 ppm silver, and 32 ppm silver) against the experimental organism Yersinia pestis, the etiological agent of bubonic fever. By applying a standard time of destruction and testing using a suspension of Y.pestis, the silver compound of the present invention was shown to be effective in the fight against bubonic plague bacteria.

[403] B. Materials and Methods [404] Y salvage, strain D27, was grown on a Columbia agar plate for 24 hours at 30 ° C in a 5% CO ₂ incubator. The grown bacteria were resuspended in 3 ml of sterile HPLC water. The suspension was transferred to a 50 ml conical centrifuge tube. The plate was then rinsed with an additional 2 mL of HPLC water. This rinse was placed in a centrifuge tube. The tube was centered at 3,500 xg for 5 minutes. The supernatant was removed but the pellet was resuspended in 1 ml of HPLC water to a final concentration of about 10 ¹⁰ cells / ml.

[405] Sequence of steps in the method [406] 1. An aliquot of 9.9 ml of the silver compound to be tested was placed in a sterile 20 mm x 150 mm tube. This tube was placed in equilibrium in a 20 ° C water bath.

[407] 2. 100 X was inserted into the tube with silver connection at the starting point of the time reference | suspensions of the test organism to form a mixture. The tube was immediately rotated and placed in a water bath again.

[408] 3. 2.min. 3min, 4min and 5min for 10 ppm or 32 ppm or 2min, 4min, 6min. and 8min for 14 ppm silver with 1.5% v / v H2O2, 1 ml of the organism / silver mixture was transferred to 99 ml of neutralizer in a 250 ml Erlenmeyer flask. The contents of the flask were thoroughly mixed.

[409] 4. The neutralized suspension was immediately serially diluted 1:10 in saline (PSS).

[410] 5. The number of viable organisms in selected tubes and flasks was determined by membrane filtration. One ml aliquots were plated on 25 ...... plates. The membranes were washed with approximately 150 ml (or 250 ml if the sample was taken from a neutralization flask) in sterile phosphate buffer and transferred to Columbia agar plates. All remaining contents (98 ml) from the neutralization flasks for 4 and 5 min were also plated. Plates were incubated at 30 ° C in a 5% CO2 incubator for 72 hours.

[411] 6. The number of colonies on each filter was counted and logarithmic reductions were calculated.

[412] C. Results [413] The results for 10 pp silver are shown in Table 7.

Table 7

Time Logarithmic reduction Destruction percent 2 min 2.63 99.77 4 min 3.20 99.94 6 min 3.46 99.97 8 min 3.68 99.98 [414] The resolved regression equation for this data is Y = 2.3965 + 0.1696x ..... ' shows that time 6 - -log reduction is 31.2 minutes. [415] The results for 32 ppm silver are shown in Table 8. Table 8 Time Logarithmic reduction Destruction percent 2 min > 7.61 99.999998 4 min > 7.61 99.999998 6 min > 7.61 99.999998 8 min > 7.61 99.999998

[416] The 14 ppm silver results are reported in Table 9.

Table 9

Time Logarithmic reduction Percentage of destruction 2 min 3.27 99.95 3 min 4.72 . 99,998 4 min 5.36 99,9996 5 min 6.47 99.99997

[417] The solved regression equation for this data is Y = 1.371+ l, 024x, shows that the time for 6 -log reduction is 4.52 minutes

This [418] Silver compound of the present invention demonstrates significantly bactericidal activity against Y.pestis, a bubonic measles ethylological agent. A 32 ppm compound gave more than 7 log reduction (actually total destruction) in less than 2 minutes. The data proves that 10 ppm silver takes about 20 minutes to achieve 6 log destruction. Silver and hydrogen peroxide show significant synergism with its 6 log destruction in less than 5 min. This is better than a single 10 ppm silver. The 14 pm silver level was chosen because data from other experiments confirmed that this silver level in combination with hydrogen peroxide could achieve the same results as the 32 ppm silver product.

[419] SUMMARY OF THE DATA [420] The following Table A summarizes the previous results on the effect of the silver compound of the present invention on a broad spectrum of microbial and human diseases. In some cases, the data in the table above is not repeated.

Either way, the results were obtained using the procedures explained above, and any middleman can effortlessly obtain these results.

[421] Human Diseases that are Cured and Pathogens Destroyed by the Silver Compound of the Invention.

Disease

Boils

Osteomyelitis

Bacterial dysentery

Burn infections

Tartar

Diarrhea (bloody)

Diaries

Ear infection

Ear infection

Typhoid

Epiglottitis (Children) Eye Infections Corneal Ulcer Disease Food Poisoning Food Poisoning

Pathogenic

Staphylococcus aureus Staphylococcus aureus Shigella boydii Pseudomonas aeruginosa Streptococcus mutans Shigella boydii Escherichia coli Haemophilus influenzae Streptococcus pneumonie Salmonella nullimurium Haemophilus influenzae Staphylococcus aureus Pseudomonas salmonella

Effective Concentration Destroyed @ 5 ppm Destroyed @ 5 ppm Destroyed @ 2.5 ppm Destroyed @ 5 ppm Destroyed @ 5 ppm Destroyed @ 2.5 ppm Destroyed @ 2.5 ppm Destroyed @ 1.25 ppm Destroyed @ 2.5 ppm Destroyed @ 2.5 ppm Destroyed @ 1.25 ppm Destroyed @ 5 ppm Destroyed @ 5 ppm Destroyed @ 5 ppm Destroyed @ 2.5 ppm Destroyed @ 2.5 ppm

Endocarditis

Endocarditis

Meningitis

Meningitis

Meningitis

Meningitis

Nosocomial infections Nosocomial infections Nosocomial infections (from hospitals)

Pneumonia Pneumonia Pneumonia Disease

Pneumonia

Respiratory tract infections Respiratory tract infections Respiratory tract infections Scarlet fever

Sepse

Cavity Infections Sinusitis Impetigo Skin Infections Skin Infections Streptococcal Pharyngitis Purulent Arthritis Throat Inflammation Urethritis (Men)

Urinary Tract Infections Urinary Tract Infections Urinary Tract Infections Urinary Tract Infections Urinary Tract Infections Vaginitis (In Women)

Wound infections Wound infections Wound infections Wound infections Wound infections Yeast infections [422] HYDROGEL EFFICIENCY

Destroyed @ 2.5 ppm Destroyed @ 5 ppm Destroyed @ 1.25 ppm Destroyed @ 2.5 ppm Destroyed @ 5 ppm Destroyed @ 2.5 ppm Destroyed @ 2.5 ppm Destroyed @ 5 ppm Destroyed @ 1.25 ppm

Destroyed @ 5 ppm Destroyed @ 1.25 ppm Destroyed @ 5 ppm Effective Concentration Destroyed @ 2.5 ppm Destroyed @ 1.25 ppm Destroyed @ 2.5 ppm Destroyed @ 2.5 ppm Destroyed @ 1.25 ppm Destroyed @ 2, 5 ppm Destroyed @ 1.25 ppm Destroyed @ 2.5 ppm Destroyed @ 1.25 ppm Destroyed @ 1.25 ppm Destroyed @ 1.25 ppm Destroyed @ 1.25 ppm Destroyed @ 1.25 ppm Destroyed @ 5 ppm Destroyed @ 10 ppm Destroyed @ 2.5 ppm Destroyed @ 2.5 ppm Destroyed @ 2.5 ppm Destroyed @ 2.5 ppm Destroyed @ 10 ppm Destroyed @ 2.5 ppm Destroyed @ 2.5 ppm Destroyed @ 2.5 ppm Destroyed @ 5 ppm Destroyed @ 2.5 ppm Destroyed @ 10 ppm

SILVER COLLIDE

Streptococcus faecalis Streptococcus gordonii Haemophilus influenzae Enterobacter aeruginosa Pseudomonas aeruginosa Streptococcus pneumonia Klebsiella pneumoniae Pseudomonas aeruginosa Streptococcus pyogenes

Staphylococcus aureus Haemophilus influenzae Pseudomonas aeruginosa

Pathogenic

Streptococcus pneumonie Streptococcus pyogenes E.coli

Klebsiella pneumoniae Streptococcus pyogenes Enterobacter aerpyogenes Hameophilius influenzae Streptococcus pneumonie Staphylococcus aureus Staphylococcus aureus Streptococcus pyogenes Streptococcus pyogenes Hameophilius influenzae Hameophilius influenzae Streptococcus mutans

Klebsiella pneumoniae Pseudomonas aeruginosa Streptococcus faecalis Enterobacter aerpyogenes Trichomonas vaginalis Enterobacter aerpyogenes Klebsiella pneumoniae Pseudomonas aeruginosa Streptococcus faecalis Candida albicans

SHAPE [423] In modern healing of wounds, it must be recognized that, for optimal healing, the wound must be sterile and protected from moisture loss and external infections. Traditional dressings are effective in protecting against external infections, but in most cases are not effective in preventing dehydration. Bandages can be given antimicrobial properties by impregnating them with various disinfectants, but these substances are often sharp and kill body cells just like microbes. Recently, a revolutionary innovation has emerged in wound care - hydrogel materials that are available in either semi-liquid (amorphous) or soft sheet-like materials. The hydrogel is hydrophilic and therefore prevents the injury from drying out. Sheet-type materials effectively prevent external infections from reaching the wound, and due to their fluid absorption capacity, the hydrogel can absorb excess fluid that is released from the injury.

[424] The hydrogel is formed by combining a hydrophilic polymer with other ingredients in an aqueous substance. The polymer forms a gel following changes in pH, temperature, or other initiating conditions. The gel's fine polymer molecular network covers ... 5 areas of water-containing substance. Although the composition may be in the form of an amorphous semi-liquid or a more stringent sheet-like material, the aqueous substance tends to occupy most of the volume, unlike the hydrophilic polymer. Hydrophilic polymers are suitable for the production of hydrogels, including gelatin, carboxide methylcellulose (and other cellulose derivatives), other vegetable or algae carbohydrate polymers such as ... alginate, carrageenan, xanthan gum, acacia gum, tragacanth gum, gum arabic resins, polymers of acrylic acid (such as corbopol), and combinations of hydrophilic and similar polymers.

[425] The aqueous component may be washed to contain various added substances that enhance the physical properties of the hydrogel and / or promote healing of wounds. These include various vitamins, amino acids, growth promoters that are added to promote healing and limit scar formation. General anesthetics such as novocaine, lidocaine and their derivatives may also be added as additives to improve the patient's well-being. Since the primary purpose of dressing is to ensure sterility of the wound, various antimicrobial and disinfectant agents are added. These include organic acids such as citric acid, dilute acetic acid, benzoic acid, and the like. Organic disinfectants include alcohol, such as isopropanol or methanol, including chlorinated phenol such as TCP (2,4,6 trichlorophenol), biguanides, chlorhexidine (mixed with cetrimide), chlorhexidine gluconate and chlorhexidine acetate. Disinfectant surfactants, which include amphoteric surfactants and aldehydes such as formaldehyde and glutaraldehyde, may be added. Effective are halogen disinfectants, which include iodine, iodoform and polyvidone iodine, as well as peroxides and other aeration agents, . Other useful ingredients are aluminum-zinc astringents, furan derivatives and quinoline derivatives such as clioquinol. However useful these antimicrobials are, they have the disadvantage that they can cause tissue damage and / or microbes can easily develop resistance to them.

[426] As explained above, the invented silver colloid is a highly effective antimicrobial agent, is very gentle to human tissues and effective against resistant microbes. Both the amorphous gel and the hydrogel sheet are capable of providing an effectively applied level of colloidal silver in a moist treatment environment. On the one hand, the amorphous hydrogel slowly releases collodium silver, which slowly enters the tissue-secreted fluid and gradually begins to dissolve. On the other hand, the amorphous hydrogel provides moisture to the tissues and at the same time helps collodial silver to reach the wound. In addition, the small amount of collodible silver in the bandage provides the advantage that, being molecular silver, it gradually decreases over time, releasing silver ions with excellent oligodynamic activity.

[427] After initial experiments, carbopol was selected as an effective hydrogel-forming agent for use with the inventive collodion silver. A base formulation containing the ingredients shown in Table 9a was created.

Table 9a

Ingredients Function Supplier Collodible silver solution (22 ppm or 32 ppm) Active, Antimicrobial and dissolution effects. Biotech Labs of America Carbopol ETD2020 Rheology modification Noveon

Triethanolamine Neutralizer, penetration agent E.Merck Propylene glycol Humidifier E.Merck

[428] All raw materials were first analyzed to determine:

1. Antibacterial activity

2. Physical and chemical properties

1. Look

2. Perfume

3. pH

4. Touching sensations

5. Density

6. Foaming ability

7. Flow [429] Collodial silver solution (22 ppm or 32 ppm):

The silver composition in this composition is used as an active component (antimicrobial agent). It is also the only solvent in this specific formulation.

[430] A. Antibacterial activity:

Culture Diameter of the zone of inhibition 22d in min Dec. 32 per min MRSA 17 mm 18mm E.coil 14 mm N / P Ps. aeruginosa 21mm 22mm

[431] B. Physical and Chemical Properties:

1. Appearance

2. Odor

3. pH

4. Background

5. Density

6. Foaming agent

7. Flowability

Colorless clear liquid

Odorless

0.5

Not applicable 1.00

Not applicable Not applicable [432] Carbopol [433] Carbopol is chemically known as carboxypolymethylene or a carboxyvinyl polymer.

It is a copolymer of acrylic acid and is a very ionic (i.e. hydrophilic) and slightly acidic compound. Carbopol polymers should be neutralized to achieve maximum viscosity. It is used in pharmaceutical, cosmetic and fabric printing applications as a thickening, suspending, dispersing, emulsifying agent. In the given formula, carbopol is used as a gelling or thickening agent.

[434] A. Antibacterial activity: Not applicable [435] B. Physical and chemical properties:

1. Appearance Dry white powder

2. Odor Odorless

3. pH Not applicable

4. Tapping Not applicable

5. Density Not applicable

6. Foaming agent Not applicable

7. Flowability Not applicable [436] Trietonalomine (TEA) C ₆ H ₁₅ NO ₃ (Mol. Weight: 149.19)

In this formulation, triethanolamine, an alkalinizing agent, neutralizes carbopol to increase viscosity. It also increases the penetration capacity of the active agent.

[437] A. Antibacterial activity (Not applicable)

Colorless viscous liquid

A little ammonia

Not applicable

Not applicable

1.1242 g / cm ³

Not applicable

Not applicable [438] B. Physical and chemical properties:

1. Appearance

2. Odor

3. pH

4. Background

5. Density

6. Foaming agent

7. Flowability [439] Propylene glycol C ₃ H ₈ O ₂ Mol.:79.09 [440] Propylene glycol is chemically known as 1: 2 propanediol. In this recipe it is used as a moisturizer and tactile modifier.

[441] A. Antibacterial activity:

Not applicable [442] B. Physical and chemical properties:

Appearance Colorless viscous liquid

2. Odor Odorless

3. pH Not applicable

4. Tapping Not applicable

5. Density 1.036 g / cm ³

6. Foaming agent Not applicable

7. Flowability Not Applicable [443] As soon as the standard formula was developed, several batches were prepared to investigate a range of possible formulations. Of the 19 experiments performed, the following observations were made.

1. Increasing the pH increases the viscosity of the gel.

2. Increasing the amount of carbopol increases the viscosity of the gel.

3. The higher the percentage of carbopol, the higher the stickiness.

From the above experiments it could be concluded that the ratio between the used carbopol and TEA and the final pH obtained should not be less than 8.5. Thus, recipe no. 18 was taken as the standard and the lot increased to 10 kg.

[444] INTRODUCTION TO PRODUCT DEVELOPMENT STUDIES:

[445] Carbopol-based gel formulations were standardized for pH, tactile, adhesive, and stability. With this in mind, various laboratory batches were used using water as the aqueous phase to obtain an appropriate quality and tactile product before the main samples were taken.

[446] Lot No. SG / 001 recipe:

Part A:. Distilled water 83.50g

Carbopol 00.62 g

NaOH 18% 00.60 g

Part B: Distilled water 1.00g

Carbopol 5.00 g

NaOH 18% 1.50 g [447] Procedure: Weigh out a quantity of distilled water from part A and keep in a water bath at 70 ° C. Add the carbopol to the distilled water, stirring constantly to avoid peaks. Add thereto NaOH 18% at 70 ° C after 20 minutes. Weigh all ingredients from Part B and hold in a water bath at 70 ° C for 15-20 minutes. Add Part B to Part A and stir for 10-15 minutes. Cool to room temperature and analyze.

[448] Results: 1.pH 10.8

2. Flow rate 90 ° C => 5 min.

45 ° C =>> 5 min.

3. Sticky Highly sticky [449] Lot No. SG / 002 Recipe:

Part A: Distilled water 83.50g

Carbopol 00.62 g

TEA 01.20 g

Part B: Distilled water l.OOg

Propylene glycol 5.0 g

TEA 1.5 g [450] Procedure: Weigh out a quantity of distilled water from part A and keep in a water bath at 70 ° C. Add the carbopol to the distilled water, stirring constantly to avoid peaks. Add TEA 18% at 70 ° C after 20 minutes. Weigh all ingredients from Part B and hold in a water bath at 70 ° C for 15-20 minutes. Add Part B to Part A and stir for 10-15 minutes. Cool to room temperature and analyze.

[451] Results: 1.pH 7.9 (SOP-08)

2. Flow rate 90 ° C => 5 min.

45 ° C =>> 5 min.

3. Stickiness Very sticky

Lot No SG / 003 Recipe:

Part A: Distilled water 86.00g Carbopol TEA 00.62g 01.20g Part B: Distilled water Propylene glycol TEA 2.00g 5.00 g 1.50 g

[452] Procedure: Weigh out a quantity of distilled water from part A and keep in a water bath at 70 ° C. Add the carbopol to the distilled water, stirring constantly to avoid peaks. Add TEA 18% at 70 ° C after 20 minutes. Weigh all ingredients from Part B and hold in a water bath at 70 ° C for 15-20 minutes. Add Part B to Part A and stir for 10-15 minutes. Cool to room temperature and analyze.

[453] Results: 1.pH 8.62

2. Flow rate 90 ° C = >> 5 min.

45 ° C = »5 min.

3. Stickiness Very sticky.

[454] Lot No. SG / 004 recipe:

Part A: Distilled water 86.00g

Carbopol 00.62 g

TEA 01.00 g

Part B: Distilled water 2.00g

Propylene glycol 5.00 g

TEA 1.50 g [455] Procedure: Weigh out a quantity of distilled water from part A and keep in a water bath at 70 ° C. Add the carbopol to the distilled water, stirring constantly to avoid peaks. Add TEA 18% at 70 ° C after 20 minutes. Weigh all ingredients from Part B and hold in a water bath at 70 ° C for 15-20 minutes. Add Part B to Part A and stir for 10-15 minutes. Cool to room temperature and analyze.

[456] Results: 1.pH 8.5

2. Flow rate 90 ° C = >> 5 min.

45 ° C = »5 min.

3. Sticky Highly sticky [457] Lot No. SG / 005 Recipe:

Part A: Distilled water Carbopol TEA 86.Og 0.62 1.20g Part B: Distilled water 2.00g Propylene glycol 7.00g

TEA

1.50 g [458] Procedure: Weigh out a quantity of distilled water from part A and keep in a water bath at 70 ° C. Add the carbopol to the distilled water, stirring constantly to avoid peaks. Add TEA 18% at 70 ° C after 20 minutes. Weigh all ingredients from Part B and hold in a water bath at 70 ° C for 15-20 minutes. Add Part B to Part A and stir for 10-15 minutes. Cool to room temperature and analyze.

[459] Results: 1.pH 8.7 2. Flowability at 90 ° C ^: =>> 5 minutes 45 ° C =>> 5 min. 3. Adhesion Very contagious [460] Lot no. SG / 006 recipe: Part A: Distilled water 85.Og Carbopol 00.62 g TEA 01:00 AM Part B: Distilled water 1.00g Propylene glycol 5.00g TEA 1.40g [461] Procedure: Weigh out a quantity of distilled water from part A and keep in a water bath at 70 ° C. Add the carbopol to the distilled water, stirring constantly to avoid from spikes. Add TEA 18% at 70 ° C for 20 minutes. Weigh all ingredients from Part B and keep in a water bath at 70 ° C for 15-20 minutes. Add B part A for part A and stir for 10-15 minutes. analyze. Cool it to room temperature and [462] Results: 1.pH 8.4 2. Flowability 90 ^° C = > 5 minutes 45 ° C =>> 5 min. 3. Adhesion Very contagious [463] Batch no. SG / 007 recipe: Part A: Distilled water 172g Carbopol 1.24g TEA 2.40g

Part B: Distilled water 6.0 g

Propylene glycol 10 g

TEA 2.80 g [464] Procedure: Weigh out a quantity of distilled water from part A and keep in a water bath at 70 ° C. Add the carbopol to the distilled water, stirring constantly to avoid peaks. Add TEA 18% at 70 ° C after 20 minutes. Weigh all ingredients from Part B and hold in a water bath at 70 ° C for 15-20 minutes. Add Part B to Part A and stir for 10-15 minutes. Cool to room temperature and analyze.

[465] Results: 1.pH 8.28

2. Flow rate 90 ° C = >> 5 min.

45 ° C = »5 min.

3. Adhesion Very sticky [466] Lot No. SG / 008 recipe:

Part A: Silver Liquid (32 parts per minute) Carbopol TEA 86g 0.62g 1.20g Part B: Silver liquid (32 parts per minute) 2.0g Propylene glycol 5g TEA 1.5g

[467] Procedure: Weigh the given amount of silver liquid from Part A and keep in a water bath at 70 ° C. Add the carbopol to the silver liquid with constant stirring to avoid peaks. Add TEA 18% at 70 ° C after 20 minutes. Weigh all ingredients from Part B and hold in a water bath at 70 ° C for 15-20 minutes. Add Part B to Part A and stir for 10-15 minutes. Cool to room temperature and analyze.

[468] Results: 1.pH 8.65

2. Flow rate 90 ° C => 5 min.

45 ° C = »5 min.

3. Adhesion Very sticky [469] Lot No. SG / 009 Recipe:

Part A: Silver Liquid (32 parts per minute) 172 g

Distilled water 12g Carbopol 1.24g TEA 2.40g Silver liquid (32 parts per minute) 6.00g Propylene glycol 10.0g TEA 2.80g

[470] Procedure: Weigh the given amount of silver liquid from Part A and keep in a water bath at 70 ° C. Add the carbopol to the silver liquid with constant stirring to avoid peaks. Add TEA 18% at 70 ° C after 20 minutes. Weigh all ingredients from Part B and hold in a water bath at 70 ° C for 15-20 minutes. Add Part B to Part A and stir for 10-15 minutes. Cool to room temperature and analyze.

[471] Results: 1.pH 8.54

2. Flow rate 90 ° C = >> 5 min.

45 ° C = »5 min.

3. Adhesion Very sticky [472] Lot No. SG / 010 recipe:

Part A: Silver liquid (32 rpm) Distilled water Carbopol TEA 172g 24g 1.39g 2.40g Part B: Silver liquid (32 parts per minute) 6.00g Propylene glycol 5.00g TEA 2.80g

[473] Procedure: Weigh the given amount of silver liquid from Part A and keep in a water bath at 70 ° C. Add the carbopol to the silver liquid with constant stirring to avoid peaks. Add TEA 18% at 70 ° C after 20 minutes. Weigh all ingredients from Part B and hold in a water bath at 70 ° C for 15-20 minutes. Add Part B to Part A and stir for 10-15 minutes. Cool to room temperature and analyze.

[474] Results: 1.pH 8.43

2. Flow rate 90 ° C = >> 5 min.

45 ° C = »5 min.

3. Sticky Sticky [475] Lot No. SG / 011 recipe:

Part A: Distilled water Carbopol TEA 24 g 0.76 g 0.56 g Part B: Distilled water 3.0g Propylene glycol 5.0g TEA 1.40g

[476] Procedure: Weigh the given amount of silver liquid from Part A and keep in a water bath at 70 ° C. Add the carbopol to the silver liquid with constant stirring to avoid peaks. Add TEA 18% at 70 ° C after 20 minutes. Weigh all ingredients from Part B and hold in a water bath at 70 ° C for 15-20 minutes. Add Part B to Part A and stir for 10-15 minutes. Cool to room temperature and analyze.

[477] Results: 1.pH 8.05

2. Flow rate 90 ° C = >> 5 min.

45 ° C = »5 min.

3. Adhesion Highly sticky [478] Lot No. SG / 012 recipe:

Part A: Distilled water 98 g

Carbopol 0.76 g

TEA 0.34 g

Part B: Distilled water 3.00 g

Propylene glycol 5.00 g

TEA 0.64 g [479] Procedure: Weigh the given amount of silver liquid from part A and keep in a water bath at 70 ° C. Add the carbopol to the silver liquid with constant stirring to avoid peaks. Add TEA 18% at 70 ° C after 20 minutes. Weigh all ingredients from Part B and hold in a water bath at 70 ° C for L5-20 minutes. Add Part B to Part A and stir for 10-15 minutes. Cool to room temperature and analyze.

[480] Results: 1.pH 6.35

2. Flow rate 90 ° C = >> 5 min.

45 ° O »5 min.

3. Stickiness Very sticky [481] Lot No. SG / 013 recipe:

Part A: Silver liquid (32 rpm) Distilled water Carbopol TEA 86 g 12 g 0.76 g 0.32 g Part B: Silver liquid (32 parts per minute) 3.00g Propylene glycol 5.00g TEA 0.64g

[482] Procedure: Weigh the amount of silver liquid and distilled water from Part A and hold in a water bath at 70 ° C. Add the carbopol to the liquid with constant stirring to avoid peaks. Add TEA 18% at 70 ° C after 20 minutes. Weigh all ingredients from Part B and hold in a water bath at 70 ° C for 15-20 minutes. Add Part B to Part A and stir for 10-15 minutes. Cool to room temperature and analyze.

[483] Results: 1.pH 6.7

2. Flow rate 90 ° C = >> 5 min.

45 ° C = »5 min.

3. Stickiness Very sticky [484] Lot No. SG / 014 recipe:

Part A: Silver liquid (32 rpm) Distilled water Carbopol TEA 86 g 12 g 0.78 g 0.32 g Part B: Silver liquid (32 parts per minute) 3.00g Propylene glycol 5.00g TEA 0.64g

[485] Procedure: Weigh the amount of silver liquid and distilled water from Part A and hold in a water bath at 70 ° C. Add the carbopol to the liquid with constant stirring to avoid peaks. Add TEA 18% at 70 ° C after 20 minutes. Weigh all ingredients from Part B and hold in a water bath at 70 ° C for 15-20 minutes. Add Part B to Part A and stir for 10-15 minutes. Cool to room temperature and analyze.

[486] Results: 1.pH 6.6

2. Flow rate 90 ° C = >> 5 min.

45 ° C = »5 min.

3. Adhesion Very sticky [487] Lot No. SG / 015 recipe:

Part A: Silver liquid (32 rpm) Distilled water Carbopol TEA 86 g 12 g 0.68 g 0.40 g Part B: Silver liquid (32 parts per minute) 5.00g Propylene glycol 7.00g TEA 0.6g

[488] Procedure: Weigh the amount of silver liquid and distilled water from Part A and hold in a water bath at 70 ° C. Add the carbopol to the liquid with constant stirring to avoid peaks. Add TEA 18% at 70 ° C after 20 minutes. Weigh all ingredients from Part B and hold in a water bath at 70 ° C for 15-20 minutes. Add Part B to Part A and stir for 10-15 minutes. Cool to room temperature and analyze.

[489] Results: 1.pH 6.72

2. Flow rate 90 ° C = »5 min.

45 ° C = »5 min.

3. Adhesive Adhesive [490] Lot No. SG / 016 recipe:

Part A: Silver liquid (32 rpm) Distilled water Carbopol TEA 86 g 12g 0.64g 0.40g Part B: Silver liquid (32 parts per minute) 5.00g Propylene glycol 7.00g TEA  0.6g

[491] Procedure: Weigh the amount of silver liquid and distilled water from Part A and keep in a water bath at 70 ° C. Add the carbopol to the liquid with constant stirring to avoid peaks. Add TEA 18% at 70 ° C after 20 minutes. Weigh all ingredients from Part B and hold in a water bath at 70 ° C for 15-20 minutes.

Add Part B to Part A and stir for 10-15 minutes. Cool to room temperature and analyze.

[492] Results: 1.pH 6.87

2. Flow rate 90 ° C = >> 5 min.

45 ° C = »5 min.

3. Adhesive Adhesive [493] Lot No. SG / 017 recipe:

Part A: Silver liquid (32 rpm) Distilled water Carbopol TEA 86 g 12 g 0.62 g 0.4 g Part B: Silver liquid (32 parts per minute) 5.00g Propylene glycol 7.00g TEA 0.6g

[494] Procedure: Weigh the amount of silver liquid and distilled water from Part A and keep in a water bath at 70 ° C. Add the carbopol to the liquid with constant stirring to avoid peaks. Add TEA 18% at 70 ° C after 20 minutes. Weigh all ingredients from Part B and hold in a water bath at 70 ° C for 15-20 minutes. Add Part B to Part A and stir for 10-15 minutes. Cool to room temperature and analyze.

[495] Results: 1.pH 7.05

2. Flow rate 90 ° C = >> 5 min.

45 ° C = »5 min.

3. Sticky Sticky [496] Lot No. SG / 018 Recipe:

Part A: Silver liquid (32 parts per minute) 86 g Distilled water 12g Carbopol 0-58 g TEA • 0.4g Part B: Silver liquid (32 parts per minute) 5.00g Propylene glycol 7.00g TEA 0.6g

[497] Procedure: Weigh the given amount of silver liquid and distilled water from part A and keep in a water bath at 70 ° C, Add carbopol to the liquid with constant stirring to avoid peaks. Add TEA 18% at 70 ° C after 20 minutes.

Weigh all ingredients from Part B and hold in a water bath at 70 ° C for 15-20 minutes.

Add Part B to Part A and stir for 10-15 minutes. Cool to room temperature and analyze.

[498] Results: 1.pH 7.40

2. Flow rate 90 ° C = >> 5 min.

45 ° C = »5 min.

3. Adhesion Smooth [499] Lot No. SG / 019 Recipe:

Part A: Silver liquid (32 rpm) Distilled water Carbopol TEA 86 g 12 g 0.54 g 0.4 g Part B: J Silver liquid (32 parts per minute) 5.00g Propylene glycol 7.00g TEA 0.6g

[500] Procedure: Weigh the amount of silver liquid and distilled water from Part A and hold in a water bath at 70 ° C. Add the carbopol to the liquid with constant stirring to avoid peaks. Add TEA 18% at 70 ° C after 20 minutes. Weigh all ingredients from Part B and hold in a water bath at 70 ° C for 15-20 minutes. Add Part B to Part A and stir for 10-15 minutes. Cool to room temperature and analyze.

[501] Results: 1.pH 7.65

2. Flowability at 9 ° C = >> 1 min.

45 ° C = »2 min.

3. Adhesion Smooth [502] Note: Although the gel pad has improved stability, it is not suitable.

[503] Based on the above results, the following instructions per batch of batch were developed.

Part A:

Silver liquid Distilled water

100 mg

860 mg

Carbopol 5.80 mg TEA 4.00 mg ASAP liquid 50.0 mg Propylene glycol 70.0 mg TEA 0.6 mg

Yield: 1.0 kg after correction for moisture loss.

[504] Procedure: In a clean sterile container, collect the required amount of distilled water and silver liquid. Raise the liquid temperature to 70 ° C with continuous stirring. Start adding carbopol in small amounts with constant stirring / homogenization. When all carbopol has been added, continue for 30 minutes. (Adjust the time according to the batch size). Then add TEA to the phase A liquid.

[505] In a separate container, mix all the ingredients of part B. Raise the temperature to 70 ° C and slowly add Part B to Part A. After complete homogenization, cool to room temperature.

[506] Caution: Carbopol dispersion should be performed using a good homogenizer. Take a small test sample using a new batch of corbapol. Reduce the warm-up time as longer warm-up leads to greater water loss.

[507] Results 1.pH 7.4

2. Flowability> 5 min.

3. Adhesion Smooth.

[508] This formulation was increased to 10 kg in a test facility. There were no problems during the magnification. Vacuuming is recommended to remove entrapped air and ensure a smooth filling.

[509] This formulation has the following physical and chemical characteristics as shown in Table 10.

Table 10

CHECK SPECIFICATION RESULTS 1. Appearance Golden yellow Transparent gel Exited

2. Odor Odorless Odorless 3. share 1.02 1.02 4. Flowability At 45 ° C and 90 ° C - More than 5 minutes to move 1 inch from the original position At 45 ° C and 90 ° C - More than 5 minutes to move 1 inch from your original location 5. Foaming agent <10 ml <10 ml 6th Touch / Stickiness 1-Smooth 1-Smooth 7th Viscosity RT 30 ° 370 32,000 ± 5,000 30,000 ± 5,000 34,000 33,500 8th PH 6.5 to 1.8 7.4 9th Freezing / thawing To exit SOP 1-10 Similar to the original 10th Optimal wavelength (X max) 22 parts per minute - 400 +/- 20 nm 32 mins per minute - 450 +/- 20 nm 400nm ** 450 nm ** 11th Insolation Without further staining Walk out. 12th Compatibility Without product decolorization by reaction with containers. Reply 3 table 13th Mitrumatdeve - 10.27% 14th Moisture absorption - 80%

[510] Microbiological Assessment [511] It is acceptable to assume that the silver colloidal hydrogel has microbiological properties similar to the original silver colloidal fluid, which has been extensively studied as demonstrated above. However, the addition of a hydrophilic polymer to form a gel could directly interfere with the microbial properties of silver, or it could slow down the diffusion of silver to reduce efficiency. Therefore, microbiological tests similar to those performed on silver colloidal fluid were also performed on silver colloidal hydrogel.

[512] Initially, the hydrogel was examined to determine if the composition was self-sterilizing. Then the following protocol was followed:

[513] Flasks with 100 ml sterile liquid thioglycolate medium (anaerobic bacterium), sterile soy casein feeding medium (aerobic bacterium) and potato-dextrose broth (fungus) were obtained. Samples for approximately palO ml gel testing were aseptically transferred to flask sets. One set was incubated at 37 ° C and the other incubated at room temperature for one week. The flasks were then examined for signs of turbidity and bacterial growth. Because the gel sample was not manufactured under sterile conditions, it can be concluded that the composition is self-sterilizing. 100 ml of gel was used for each assay. This corresponds to 2; 2pg per 100 ml medium or 0.02214 or 0.032pg silver per ml medium. At this concentration, silver will have no antimicrobial activity and can thus eliminate the negative effects.

[514] Various test organisms were then used to compare the zone of inhibition achieved with 22 or 32 parts per minute. of silver liquid or 22 or 32 parts by min. silver gel prepared as described above and shown in Table 11. Aliquots of 0.1 ml or vigorously growing 18 h of each micro-organism. cultures (about 10 ⁸ coliform units / ml) were plated on sterile nutrient agar plates. A 10 mm hole was made in each inoculated plate using a cork drill. The assay volume of product (0 / 2-0.3g) was placed in each well and the plate was incubated for 24 h. The following zones of inhibition were then examined and measured.

[515] Table 11

Cult Silver liquid Sudra a gel 22 parts per minute 32 parts per minute 22 parts per minute 32 parts per minute E.coil 14 mm 14 mm 12 mm 13 mm Ps. Aeruginosa 21mm 22mm 21mm 20 mm B. subtilis 15 mm 16 mm 14 mm 14 mm MRSA 1 17 mm 18mm 16 mm 17 mm MRSA 2 16 mm 17 mm 16 mm 17 mm S. aureus ATCC 6538 P 14 mm 14.5mm 15 mm 15 mm Spyogenes 16 mm 18mm 16 mm 18mm S.typhi 17 mm 16 mm 16 mm 16 mm Sh. flexneri 20 mm 21mm 20 mm 21mm K. pneumoniae 17 mm 18mm 18mm 18mm C.diptheriae 16 mm 18mm 16 mm 17 mm C.albicans 39 mm 40 mm 39 mm 40 mm

[516] These results show that the effects of gel inhibition are essentially equivalent to those of silver colloidal fluid; it demonstrates that the gelling polymer does not adversely affect the antimicrobial capacity of the silver colloid. Some cultures (S. Pyogenes, C. Diphthrise and S. Aureus) were also cultured on blood agar. The results show that the silver gel will also be effective on a bleeding or fluid-wound.

[517] Similar tests were performed on the same bacterial genus using different antibiotic agents. In some cases, antibiotics were more effective than silver compounds - in others, much less effective. This demonstrates that the strains used were not attenuated (See Tables 12a and 12b).

Gram-positive bacteria (Table 12a)

Antibiotics Conc. S. aureus MRSA1 MRSA2 B. subtilis Ampicillin 200 mg Clean 15 mm 18mm 16 mm Cefotaxime 30 mcg 26 mm Are not inhibition Clean 12 mm Cephalexin 30 mcg Clean 1.0 mm 0.8 mm Clean Ciprofloxacin 5mg 28mm 14 mm 14 mm 20 mm Cloxacillin 1 mcg Clean Clean 13 mm 18mm Cotrimoxazole 25 mcg Clean Are not inhibition Are not inhibition 15 mm Gentamicin 10 mg Clean Are not. inhibition 11 mm 18mm Lincomycin 2 mcg Clean Clean Clean 18mm Ofloxacin 5mg Clean 15 mm 16 mm 22mm Pefloxloxacin 10 mg 30 mm 11 mm 13 mm 21mm Roxithromycin 15mg Clean 1.0 mm 12 mm 20 mm Tetracycline 30 mcg 34 mm Are not inhibition 0.7 mm 19mm

Gram negative ? acres (Table 12b) Antibiotics Conc. E.coli K. pneumoniae S.typhi Ps. aeruginosa Amikacin 30 mcg Clean 18mm Clean 10 mm Ampicillin 200 mg 23mm 18mm 20 mm 13 mm Cefotaxime 30 mcg 21mm 20 mm 22mm 19mm Ceftizoxime 30 mcg 18mm 18mm 15 mm Are not inhibition Chloramphenicol 30 mcg 22mm 21mm 23mm Are not inhibition Ciprofloxacin 5mg 29 mm 22mm 25 mm 15 mm Co-trimoxazole 25 mcg 24mm 19mm 27mm Clean Gentamicin 10 mg Clean 17 mm Clean Are not inhibition Ofloxacin 5mg Clean 29 mm Clean 15 mm Pefloxacin 10 mg Clean 25 mm Clean 10 mm Piperacillin 100 mg 22mm 15 mm 16 mm 10 mm

Tetracyclic 30 mcg 19mm 18mm 16 mm Are not inhibition

[518] Hand Scrap Examination [519] Because the hydrogel is capable of increasing the adhesion of silver to the skin, the effectiveness of the gel was evaluated as a hand scrub. For this test, a one-inch square was marked on a volunteer's arm and then shelled with 1 g of gel. The control field was shelled with sterile distilled water. The fields were wiped with a swab and a strip was applied to the nutrient agar with a swab. The swab was repeated every hour for four hours. Plates with stripes were incubated for 24 h. at 37 ° C and the results were evaluated.

[520] As shown in Table 13 below, so many bacteria grew on control swabs that they were too numerous to count (PDLS). The fields treated with silver gel remained sterile for three hours and showed only slight growth within four hours. They should provide the perfect result for health care professionals who need to sterilize the surface of their hands without the use of coarse or moisturizing compounds.

Table 13

Time Control 22 parts per minute 32 parts per minute 0 hr PDLS No growth No growth 1 hr PDLS No growth No growth 2 hours PDLS No growth No growth 3 hours PDLS No growth No growth 4 hours PDLS 3 collections 2 collections

Although hydrogels exhibit excellent wound healing properties, a typical disadvantage of hydrogels is that microorganisms are often able to migrate through the matrix. That way, if the injury is covered with a hydrogel and one part of the injury is infected, the infecting organisms will be able to move through the hydrogel and infect other areas. This possibility was tested using a hydrogel strip to connect the separated areas on a nutrient agar plate. Each agar plate was divided into two zones by removing a 2 cm agar strip along the plate diameter. This gap was connected to a 1.5 cm wide strip that partially entered the agar to about 5 mm at each end. One side of the plate was then inoculated with about 0.5 ml of culture and the plate incubated to see if microorganisms could cross the hydrogel "bridge". The results in Table 14 show that silver hydrogel completely prevented migration.

Table 14

Cult Inoculation zone Migration zone E.coil Strong growth No growth B.subtilis Strong growth No growth MRSA 1 Strong growth No growth Ps. In Aeruginos Strong growth No growth Hydrogel control Strong growth Growth

From the results shown above, the gel formula of the prototype was selected and variations are presented in the following examples.

[521] Example Batch of A-Gel Take the components of Part A and Part B as shown below:

Part A: Patented silver colloid 32 parts per min.860 g

Distilled water 100 g

Carbopol 6.8 g

Triethanolamine 4.0 g

Part B: Patentable silver colloid 32 parts per minute 50g

Propylene glycol 70 g

Triethanolamine 6.0 g [522] First, take the required amount of distilled water and silver liquid into the mixer and start mixing. Add corbapol slowly (Noveon, USA). The stirring should be strong enough to dilute the carbopol and avoid the formation of peaks. The temperature of the mixing should be maintained at 60-70 ° C.

[523] Mix all the ingredients of Part B in a crusher. Bring to 70 ° C and add to part A, stirring vigorously. Continue stirring and cool to room temperature. Check the outcome of the batch. It should be about 1000 gm. Triethanolamine causes the carbopol to be converted into a gel.

Example B:

[524] Prepare all ingredients as in Example A, including the addition of 1% collagen. This will give the gel both antimicrobial and collagen properties, which have the effect of accelerating the treatment of Turat type injuries.

Example C:

[525] Prepare all ingredients as in Example A but including woody aloe (powder or liquid) in the range of 1-5%. This will give additional wound healing properties.

Example D:

[526] Prepare all ingredients as in Example A by adding 1-10% maltodextrin by weight. This will give a gel formula that stimulates the granulation of the wounds.

[527] Summary of silver hydrogel results [528] Carbopol-based gels were prepared using patented silver colloidal liquids at 22 ppm. and 32 parts per minute Gels made in this way have many advantages over their analogues, thanks to their ability to stay in place while retaining the properties of the mother's silver fluids. The medicinal nature of the amorphous hydrogel provides the benefits of accelerating the treatment of wet injuries and also limits the severity of burn injuries by reducing heat shock. In addition, the active agent, colloidal silver fluid, was studied on a cell line in an earlier study and it appears to be non-cytotoxic.

[529] A thorough physico-chemical evaluation of the gel was carried out in several batches and a series of detailed methods were prepared to standardize and control the product and processes during production.

[530] Thorough microbiological studies have been performed which show that the gel retains its bactericidal properties. Simulated silver movement studies show that the gel is capable of supplying the wound with silver for a long time. It is designed to prevent germs from entering the wound.

[531] Research has shown that in a hypothetical evaluation of silver hydrogel in a publication (12th issue of Wound Care Journal, September 8, 2003) that also deals with other silver-based gels, it has been awarded points for:

1. Microbial suppression in the wound area;

2. Microbial killing tests;

3. Microbial migration; and

4. Silver content in gels.

[532] The first silver hydrogel test is for group "B" and the other three tests are for group "A" with a total score of 20. Commercial products such as Calgitrol Ag and Acticot have also received an equivalent score.

[533] Due to the antimicrobial and antiviral properties of the colloidal silver solution, the gel can not only be used as an ointment for wounds. As described above, Hydrogel is a great antimicrobial scrub for hands. In addition, due to its non-irritating properties, the combination of silver colloid and hydrogel is a great personal lubricant for women and men for sexual purposes that can be used with condoms or diaphragms. This combination combats bacteria, fungi (a remarkable effect on Candida albicans) as well as HIV by disinfecting reusable precautions such as diaphragms. Unlike other personal lubricants, the hydrogel does not have a harmful effect on condoms or diaphragms because of its low oil content (if any).

[534] HYDROGEL LIN HAND WASH detergent (Note: Hydrogel and SILGEL are the same product name used below). resistance in health care. In accordance with MMWR Issue 51, October 25, 2002 RR-16, an efficacy test of the SILGEL hand hygiene product was performed.

The following standard exposure procedure was performed:

Required Material (SOP):

Serratia marcescens (10 ⁸ cfu / ml) standard suspension, running water, sterile rubber gloves,

Sterile sample solution, sterile tryptase soy agar, sterile pipettes and sterile tubes.

Method:

1. Apply 5 ml of standard suspension of Serratia marcescens to your hands.

2. Apply 3 ml of the test material to the forearms.

3. Pour 3 ml of water onto the hands (see Figure 1).

4. Wash hands and forearms in room temperature water for 30 seconds.

5. Repeat procedure 2 to 4.

6. After the first, third, seventh and tenth washes, put on sterile rubber gloves.

7. Pour 75 ml of the sample into the gloves.

8. Massage hands for one minute.

9. Samples are obtained for septic analysis by qualitative analysis. Use tryptase soy agar.

10. Work with tube coating techniques using the original, ΙΟ ′ ¹ , 10 ′ ² , and 10 ′ ³ as solutions.

The tubes are incubated at 37 ° C for 24 hours.

MATERIALS AND METHODS

The above SOP procedures have been used

Product used: Sterile tryptase soy agar.

Cultures used: 16-hour-old culture of Serratia marcescens (approximately 10 ⁸ CFU / ml)

Incubation temperature: 37 ° C

Incubation time: 24 hours

Rated products: Silgel 22 and 32 ppm, Spitaderm, Liquid clean, Sterilium

Results: The results are shown in Tables 15a-15e, 16a-16e

Left hand i 15a. Table: Spitaderm (Annex II)

Number of washes V Solution CFU / ml The original 10 ' ¹ IO ' ² 10 ' ³ 1.wash 10th Zero Zero Zero 3. washing Zero Zero Zero Zero 5. washing Zero Zero Zero Zero 10. washing Zero Zero Zero Zero

15b. Table: "Sorghum" 32 ppm

Number of Wash Solution CFU / ml The original 10 ¹ IO ' ² 10 ' ³ 1 washing Zero Zero Zero Zero 3. washing Zero Zero Zero Zero 7. washing Zero Zero Zero Zero 10. washing Zero Zero Zero Zero

15c. Table: "Sorghum" 22 ppm

Number of Wash Solution CFU / ml The original IO ' ¹ 10 ' ² 10 ' ³ 1.wash Zero Zero Zero Zero 3. washing Zero Zero Zero Zero 7. washing Zero Zero Zero Zero 10. washing Zero Zero Zero Zero

15d Table: "Sterillium" (Annex III)

Number of Wash Solution CFU / ml The original 10 ' ¹ 10- I0 ' ³ 1 washing 30th 30th Zero Zero 3. washing Zero Zero Zero Zero 7. washing Zero Zero Zero Zero 10. washing Zero Zero Zero Zero

15e. Table: Liquid clean (Annex III)

Number of Wash Solution CFU / ml The original 10 ' ¹ 10 ' ² 10 ' ³ 1.wash Zero Zero Zero Zero 3. washing Zero Zero Zero Zero 7. washing Zero Zero Zero Zero 10. washing Zero Zero Zero Zero

Right hand

16a. Table: Spitaderm (Annex II)

Number of Wash Solution CFU / ml The original 10 ' ¹ 1 (P 10 ' ³ 1.wash Zero Zero Zero Zero 3. washing Zero Zero Zero Zero 5. washing Zero Zero Zero Zero 10. washing Zero Zero Zero Zero

16b. Table: "Sorghum" 32 ppm

Number of Wash Solution CFU / ml The original 10 ' ¹ 10 ' ² 10 '^ 1 washing Zero Zero Zero Zero 3. washing Zero Zero Zero Zero

7. washing Zero Zero Zero Zero 10. washing Zero Zero Zero Zero

16c. Table: «Silgel» 22 ppm

Number of Wash Solution CFU / ml The original 10 ' ¹ 10 ' ² 10 ' ³ 1.wash Zero Zero Zero Zero 3. washing Zero Zero Zero Zero 7. washing Zero Zero Zero Zero 10. washing Zero Zero Zero Zero

16d. Table: «Sterillium» (Annex III)

Number of Wash Solution CFU / ml The original 10 ' ¹ no 10 ' ³ 1.wash Zero Zero Zero Zero 3. washing Zero Zero Zero Zero 7. washing Zero Zero Zero Zero 10. washing Zero Zero Zero Zero

16e. Table: Liquid clean (Annex III)

Number of Wash Solution CFU / ml The original 10 ' ¹ 10 ' ² 10 ' ³ 1.wash Zero Zero Zero Zero 3. washing Zero Zero Zero Zero 7. washing Zero Zero Zero Zero 10,. washing Zero Zero Zero Zero

Conclusion: SILGEL 32 ppm and SILGEL 22 ppm meet the efficacy criteria for EGM (Experimental End Monograph) Hand Hygiene Products, which indicates the efficacy of 2 logi9 for reducing indicator organisms on each hand after first use and 3 logio for reducing indicator organisms on each hand within 5 minutes after

10.use of use. SILGEL proved to be more effective than Sterilium and Spitader. Because SILGEL is hand-washable, it is easy to use because it does not need to be rinsed. It is noted that it does not dry the skin and does not cause irritation and moisturizes the applied skin.

[536] Hydrogen Gel as a Wound Care Product [537] Introduction [538] Hydrogel agents can be used as primary dressings (amorphous and impregnated gauze) or as primary or secondary dressings (sheets) to process partial, large and deep (amorphous and amorphous) ), as well as necrotic wounds, necrotic tissue, second degree burns and radiation damage.

[539] Almost all hydrogels available on the market do not contain antimicrobial substances. This is because antibiotics and antiseptics are potentially cytotoxic and often impede wound healing.

Since the inventive silver / water solution is not cytotoxic, it was decided to prepare it as a hydrogel using the developed nanoparticles.

Recently made specially formulated hydrogel for the treatment of radiation-induced dermatitis. These care products have high, specific temperatures that produce a cooling effect and absorb at least three times more water, serum or blood.

[540] Advantages • Soothing effect, soothes pain • Moisturizing wounds • Promotes autolytic debridement • Fills base (amorphous and impregnated cotton wool) • Provides minimal to moderate absorption • Easy to apply and cleanable from wound • Can be used for infective 54 ] Disadvantages • Not recommended for high wound wounds • Occasionally secondary dressings may be required • Slightly dehydrates if not covered • May be difficult to apply • May cause maceration [542] Procedure [543] The currently available hydrogel is prepared as a dressing wounds. The shape of the hydrogel sheet is called SILDERM.

[544] Results [545] SILDERM - LOSS OF MOISTURE [546] Purpose:

• Determine SILDERM Moisture Loss Capacity [547] Procedure [548] Required Features:

• Analytical balance.

[549] Required Materials:

• Plastic tray [550] Method:

• Determine the weight of the empty tray.

• Place the SILDERM bandage on the tray.

• Determine the total weight of the tray and SILDERM.

• Mark reading as t = 0 hours.

• Read every hour.

• Take 24-hour readings.

• Make a chart of time and moisture loss.

• Determine the percentage of moisture loss.

Results: See Table 17 and Figure 34

Table 17: SILDERM Moisture Loss Capacity

Time (hours) Weight 0 68 gm 1 64 gm 2 60 gm 3 56 gm 4 51 gm 5 48 gm 22nd 40 gm

Conclusion: It can be concluded that SILDERM bandage can lose up to 30% of its weight.

[551] SILDERM - MOISTURE CAPACITY [552] Purpose:

• Determine the absorption capacity of the dehydrated SILDERM.

[553] Procedure:

• Analytical balance.

[554] Resources needed:

• Beaker.

[555] Method:

• Determine the weight of the SILDERM bandage in grams.

· · Mark reading as t = 0 hours.

• Place the SILDERM bandage in the beaker, soaking it completely.

• Remove the bandage every hour and determine its weight.

• Take 24-hour readings.

• Determine the percentage of moisture absorption capacity.

Results: (see Table 18 and Figure 3 5)

Table 18

Time (hours) Weight 0 45 gm 1 48 gm 2 51 gm 3 53 gm 4 54 gm 5 56 gm 6th 57 gm 22nd 68 gm

Conclusion: The dehydrated SILDERM bandage can absorb up to 52% moisture.

[557J SILDERM - SILVER CASE [560] Purpose _ · To determine the long-term yield of SILDERM silver nanoparticles [561] Principle:

• Hydrogel dressings are usually applied to the wound for 48 to 72 hours. In this case, it is necessary to determine the antimicrobial activity of the bandage by determining the silver yield during the reference period.

[562] Resources needed:

• · Incubator, laminate heat transfer.

[563] Resources needed:

• Sterile nutrient agar plates, sterile cotton swabs, micropipette (100 - 1000 μΐ capacity), 16-hour Pseudomonas aeruginosa culture (wild) [564]

• Cut off a 4 x 3 cm piece of SILDERM bandage.

• Place the SILDERM dressing on a plate impregnated with Ps. aeruginosa (wild) • Incubate for 18 hours at 37 ° C.

• Examine the exposure area vertically and horizontally.

• Place the same SILDERM dressing on a nutrient agar plate freshly impregnated with Ps. aeruginosa • Incubate as described above.

Repeat the procedure for at least 7 days.

Results: Throughout the experiment, SILDERM showed the properties listed in Table 19 in 3 runs.

Table 19 SILDERM parse test

Section number Vertical movement Horizontal movement Step 1 52mm 35 mm Step 2 53 mm 35 mm Step 3 51mm 34 mm

Conclusion: SILDERM hydrogel is capable of antimicrobial activity in 3 runs every 24 hours. Further testing in progress.

[565] The content of the features for the above process is as follows:

Nourishing Agar:

Peptone Sodium chloride Meat piece Distilled water Agar pH

10.0 gm 5.0 gm 3.0 gm 900 ml 2.5 gm 7.2 ± 0.2 [565] You will be able to formulate SILDERM with the following materials:

• Collagen

Collagen, the most common protein in the body, is fibrous and insoluble produced by fibrobacilli. Its fibers are found in connective tissues, including the skin, bones, ligaments and cartilage. In wound healing, collagen promotes the formation of newly formed collagen fiber granulation tissue at the base of the wound, creating an environment conducive to healing.

• Maltodextrin

Maltodextrin is a wound healing agent that accelerates the activation and attraction of macrobacteriophages, thus limiting the sources of infection and promoting granulation.

• Derivative Platelet Derivation Factor (ATVF) ATVF promotes the chemically-tactile attachment and formation of cells necessary for wound healing and granulation tissue formation. It is mainly used in the treatment of the lower parts in the treatment of diabetic neuropathic ulcers.

[567] Disodium ethylene ditamine tetra acetic acid (EDTE) as an additive [568] Disodium EDTA promotes antimicrobial activity in natural and synthetic compounds by such mechanism. Its mechanism of action is believed to promote cell permeability, thereby facilitating access to antimicrobial compounds.

[569] Metal chelator and bacterial outer membrane permeation enhancer EDTE activity enhances the activity of various antimicrobial agents acting on Pseudomonas i aeruginosa. Addition of a concentrate of EDTE subinhibitors significantly reduces cefprozil microbes against E. coli and Serratia marcescens.

[570] Imipenem, ceftazidime and cefipime plus 150 mcg EDTA have been found to increase the diameter of the P. aeruginosa suppression zone. Studies show that ethylene ditamine tetra acetic acid (EDTE) affects the susceptibility of P. aeruginosa. The use of EDTE in combination with AgNOj significantly enhances the antimicrobial activity, resulting in Klesbiella pneumoniae and Staphylococcus aureus, resistant to 70 micrograms / ml AgNCI, sensitive to 10 micrograms / ml of this composition.

A specific potion was developed and several tests were developed to determine the efficacy of the currently available silver / water composition with disodium EDTE. Disodium EDTE was obtained from the West Bank of Mumbai in India. Disodium EDTE, also known as Na 2 EDTE (disodium ethylene ditamine tetraacetic acid), has the following formula: (CH ₂ N (CH ₂ COOH) CH 2 COONa) _{2 2} H ₂ O. It has a molar mass of 372.24.

[571] Agents used in this test: nutrient agar: (HiMedia) 1000 ml; No. 1G115, Expires August 2006, Animal tissue peptic digestion 50.00g; yeast extract 1.50g; beef 1.50g, sodium chloride 5.00g, agar type -125g; pH 7.4 ± 0.2.

Antimicrobial activity ppm silver / water composition alone, 22 ppm silver / water composition alone, and 32 ppm silver / water composition alone, as well as 22 ppm silver / water composition alone were added to Na ₂ EDTE and tested against microorganisms, including:

Escherichia coli (resistant to many drugs) from stool.

Pseudomonas aeurginosa (many medicines resistant to sputum)

Methicillin-resistant Staphylococcus aureus (many drug-resistant) pus in the pelvis.

The above samples were obtained from P.D. Hindu Hospitals (Mumbai, India).

Shigella flexneri (laboratory sample)

Salmonella typhi (laboratory sample)

Bacterial samples were stored at 37 ° C for 24 hours on nutrient agar (pH 7.4).

[572] 32 ppm and 22 ppm solutions of Na2 EDTE were prepared in sterile distilled water. Each microorganism was dissolved in sterile salt water in 10 ⁶ colony forming units (cfu / ml). They were placed on top of nutrient agar (pH 7.4) with sterile cotton swabs. Wells (10mm diameter) were plated in the agar and injected with 0.1 ml volumes of the respective solution. After about 24 hours of incubation at 37 _° C degrees, all plates were monitored to assess the progress and extent of the diameter was measured in a zone reader (Hi Media). The results are shown in Table 20.

Results and discussion

Table 20 Silver / water + Na ₂ DETE

The system E.coli MRSA C. albicans

Silver / water 32 ppm-control 21mm 24mm 27mm Silver / water 32 ppm + 0.5% Na ₂ EDTE 22mm 29 mm 40 mm Silver / water 22 ppm + 20 mm 23mm 29 mm Silver / water 22 ppm + 0.5 Na ₂ EDTE 22mm 31 mm > 40 mm

[573] Disodium EDTE at 0.5 ppm significantly advances the potential of silver / water compounds at both 22 and 32 ppm concentrations.

[574] Silver EDTE as a separate antimicrobial agent [575] A specific compound and test set was developed to determine the antimicrobial properties of silver chelation such as silver EDTE (or AgEDTE). The following commercially available silver EDTE compositions were studied: ,, Nobel and Alpha Chemicals.

[576] Resources needed:

• Incubator, laminar heat transfer.

[577] Required Materials:

• Sterile nutrient agar plates, sterile cotton swabs, micropipette (100 - 1000 μϊ capacity), 16-hour culture consisting of the following bacteria (approximately 10 ⁸ CFU / ml):

Escherichia coli (wild), Escherichia coli (MDR), Pseudomonas aeruginosa (wild), Pseudomonas aeruginosa (MDR), Staphylococcus aureatus ATCC 6538P, methicillin-resistant Staphylococcus aureatus.

[578] Method:

• Place the 16-hour-old culture on a sterile nutrient agar plate.

Allow to soak for 15 minutes.

• After 15 minutes, aseptically create wells on the agar surface with a 10 mm stopper.

• Inject 100 μΐ of the sample solution into the wells. Leave to soak for 15 minutes.

• Incubate plates at 37 ° C for 24 hours and evaluate results.

• Measure size with a zone reader (Hi Media) to evaluate development and diameter.

[579] Results: See Table 21, Figures 36 and 37.

[580] Table 21: Comparative evaluation of silver chelators [581]

The organism Concentration Exposure area AKZO ALPHA E. coli (wild) 28 ppm 20 mm 22mm 57 ppm 22mm 24mm 114 ppm 22mm 25 mm E. coli (MDR) 28 ppm 20 mm 19mm 57 ppm 21mm 21mm 114 ppm 23mm 22mm Ps. aeruginosa (wild) 28 ppm 21mm 17 mm 57 ppm 27mm 24mm 114 ppm 28mm 27mm Ps. aeruginosa (MDR) 28 ppm 15mm 17mm 57 ppm 21mm 20 mm 114 ppm 25 mm 22mm S. aureus 28 ppm 16 mm 15 mm (wild) 57 ppm 19mm 18mm

114 ppm 22mm 21mm MISA 28 ppm 19mm 20 mm 57 ppm 21mm 22mm 114 ppm 26 mm 24mm

[582] Conclusion: Silver chelators such as silver EDTE have antimicrobial activity.

[583] ANTIBIOTIC COMBINATION THERAPY [584] When antibiotics were invented, they were considered a miracle cure to some extent. Infections that were fatal in the past are now considered minor inconveniences. Medicine seems to have gone full circle. Misuse, misspelled prescriptions, and / or misuse of antibiotics have led to the emergence of new types of microbes and new health threats.

[585] Some other factors that make bacteria immune are the use of antibiotics for agricultural purposes and as food supplements for agriculture (such as birds, cattle, pigs, etc.). Incorrect antibiotic prescription has affected the agricultural industry in the US and many other countries. Antibiotic therapy is often initiated before the specimen is delivered to the laboratory. Avian flu (such as H5N1 or "SARS") is highly resistant to antibiotics due to its very high daily use by Asian farmers. Patients have easy access to antibiotics 'over the counter'. Incorrect dosage and incomplete treatment can also cause bacterial immunity. Many drugs, including penicillin, were considered miraculous and initially had great potential for effective control. Later bacteria have significantly limited the use of the drug.

[586] Antibiotic resistance is a global problem nowadays. Some common bacterial species, such as the staphylococcus aureus foci found in hospitals, are immune to vancomycin. MISA (methicillin-resistant Staphylococcus aureus) causes acute nosocomial infections, which often results in hospital closures and even destruction.

[587] In the event of such problems, alternatives must be sought. One alternative is to look for new antibiotics and replace the old ones. Another alternative is to make existing antibiotics more effective. Increasing danger is caused by bacteria that are immune to multiple drug types, so the effects of silver / water compositions should be seriously considered.

[588] One way to deal with the problem is to use combination therapy that uses two or more types of antibiotics with different effects. There are several types of synergistic antibiotics, but the results of the studies may show signs of incompatibility and the use of such combinations does not exclude the development of immunity.

[589] Aims and Objectives [590] This study was conducted with the following aims and objectives:

1. To determine the multi-antibiotic immunity (OR) model in clinical isolates.

2. Determine the sensitivity of clinical isolates to silver / aqueous solutions.

3. Determine antibiotic combination (synergy) after disk roughness test.

4. Determine the minimum inhibitory concentration (MIC) for antibiotics and silver / water solution.

5. Investigate the effects of synergistic antibiotics and silver / water solutions on a chessboard test basis.

[591] MATERIALS AND METHODS

Collection of clinical isolates

The following clinical isolates of several antibiotic immunity were obtained from Hindu Hospital

Kadel Road, Mahima, Mumbai - 400016, India.

• · Escherichia coli (isolated from stool) • Pseudomonas aeruginosa (isolated from sputum) • Methylycin resistant Staphylococcus Aureus (MISA - Isolated from purulent pelvic area) [592] Agents, solutions and antibiotic discs:

[593] Features:

• Nourishing fluid.

• Nourishing agar.

• Muller and Hinton agar.

[594] Solutions:

• Antibiotic solutions.

• Silver / water solution (22 ppm).

The agents and solutions used in the experiments are listed in Table 26 (below).

Used ready made available antibiotic discs of appropriate concentration. The contents of the discs are shown in Table 27 (below).

Inoculum preparation:

The culture medium was inoculated and left for incubation. 37 ° C overnight. 500 mL of overnight cultures were transferred to 5 mL of new nutrient solution and placed for re-incubation for 4 to 6 hours at the same temperature. The culture density is approximately 10 ⁵ to 10 ⁶ cfu / ml.

[595] Antibiotic Sensitivity Test - Kirby Bauer method:

In this method, antibiotic-impregnated disks were placed on an agar plate that had previously been inoculated with a bacterial suspension. Antibiotics are spreading in the environment.

As the distance from the disk increases, allogarithmic reduction occurs in the antibiotic concentrate.

A clean area is formed around the disk, indicating the susceptibility of organisms to antibiotics.

The clean areas are measured in millimeters and compared with the standard NCCLS table.

.1 [596] Method:

1. Sterile cotton swabs were dipped in the nutrient fluid described above and plated on agar plates to observe convergent development.

2. The inoculum was left to soak and the antibiotic discs were placed on the surface with sterile surgical forceps.

3. The plates were left for incubation at 37 ° C for about 24 hours.

4. The appearance of a cleaned area around the disk indicates the sensitivity of organisms. The diameters of the squares are fixed and marked according to the standard tables provided by NCCLS.

(See Table 27) (Koneman, Issue 5, 1997) [597] Sensitivity of Isolates by 10 ppm Agar Diffusion Method:

It was determined by a test method in which an isolated volume was injected into the agar and 10 ppm silver / water solution was added to the wells (10 mm). The size of the area is then determined.

[598] Method:

1. Add 0.5 ml of inoculum to melted Muller and Hinton agar (20 ml), place on a petri dish and leave to solidify.

2. The holes in the agar layer are pierced.

3. Solutions of varying degrees of silver / water concentration are introduced into each of the pierced holes.

4. Leave the plates to incubate at 37 ° C for approximately 24 hours.

5. Areas of exposure are recorded.

[599] Detection of Antibiotic Combinations by Diffusion Diffusion Test.

This is a simple, high-quality test for determining the combination of a clinical isolate and an antibiotic. In this test, antibiotic disks are placed on an agar plate inoculum by the Kirby Bauer method. The discs are placed at a distance slightly larger than the diameter of the area of exposure of each disk. The shape of the site of action indicates the interaction between the clinical isolate and the combination of antibiotics.

[600] Method:

1. Immerse sterile cotton swabs in the above culture medium and grease with M.H. agar plates to achieve convergent development.

2. After infiltration of the inoculum, 2 antibiotic discs are placed on the surface (combination studied) with sterile surgical forceps. The discs are placed at a distance slightly larger than the diameter of the area of exposure of each disk.

3. Leave the plates to incubate at 37 ° C for approximately 24 hours.

4. The shape of the site of action indicates the interaction between the clinical isolate and the combination of antibiotics.

Figure 25 shows a table showing the bacterial interaction of disk diffusion.

[601] Part A shows additive or insensitivity effects; each antibiotic has its own zone of action not affected by other antibiotics. Part B lists the opposite effects of reducing the effect of one antibiotic on another. Part C shows two possible manifestations and a synergistic interaction. On the left side, there is a larger exposure area where two antibiotics collide. On the right side, the antibiotic itself is not effective, but in combination with another it becomes effective against bacteria.

[602] Determination of Minimum Effective Concentration (MIC) for antimicrobial agents. This is a cloud-based susceptibility test. Serial solutions of the antimicrobial agent are prepared and added to the standardized bacterial suspension. At the end of the incubation period, development in all test tubes used is tested. The lowest concentration of antimicrobial agent that has a marked effect on development is considered to be the basis.

Antibiotics used:

Amikacin: Mycacin inj. (250 mg) Aristo Labs, Mumbai, India.

Lot No 02D054, exc. April 2004.

Cefoperazone: Magnamycin Inj. (250 mg) Pfizer Ltd., Mumbai, India

Lot No 32035153A, exc. March 2003.

Ciprofloxacin: Cyphran (200mg / ml) at Ranxbay Labs, Jaipur, India

Lot No 9042601, exc. March 2004.

[603] Method:

1. Doses of the antimicrobial agent are administered in an appropriate medium.

2. An antimicrobial agent is used to control development in a sterile tube.

3. A standard bacterial suspension is inoculated into each tube and left for incubation at 37 ° C for 24 hours.

4. At the end of the incubation period, the turbidity in each tube is thoroughly examined. Troubles indicate that the antimicrobial agent has not inhibited bacterial growth.

5. MNC is at the lowest concentration required to obviously inhibit bacterial growth.

[604] Study of synergistic activity on the chessboard test principle.

The chess board test principle is applied when working with several antibiotics and / or solutions. A number of double solutions are selected so as to include concentrations ranging from one sixteenth to at least twice the dose of MNK. Medicinal A is dissolved in the ordinate and B in the abscissa. The resulting chessboard table indicates all possible combinations of two antibiotics from the tube with the highest combination in the opposite corner.

Minutes

2x

MNK

ΜΝΚ

1: 2

ΜΝΚ

1: 4

ΜΝΚ

1: 8

ΜΝΚ

1:16

ΜΝΚ

Ο

+ ve C

1:16

ΜΝΚ

1: 8

ΜΝΚ

1: 4

ΜΝΚ

1: 2

ΜΝΚ

ΜΝΚ

2χ

ΜΝΚ ,, Β ”drug pg / ml / ,, Α” drug pg / ml

Chairs table layout.

The first row and column indicate the presence of only one drug per tube to demonstrate individual antibiotic MNC values in the test isolates.

One tube without antibiotics is stored for control.

1. Final volume - 5 ml is injected into each tube and dissolved in the appropriate medium.

2. Add 0.1 ml of culture suspension.

3. Leave to incubate at 37 ° C for 24 hours.

4. The results are represented as an isobologram, with dots for each type of combination having the same effect, including equally effective concentrations for each antibiotic individually.

[605] Calculations:

Eliot and others (1954) described a way to determine the amount of MIC in the results obtained by fractional inhibitory concentration (FIC) index, defined as the sum of the FIC values of two drug combinations.

FIC index = FIC for medication A + FIC for medication B.

Medication A FIC = MIC for medication A in combination with medication B for medication A

An index lower than 0.5 is considered to be a synergy syndrome index greater than 2.0 indicating antagonism.

(Koneman, Vol. 5, 1997).

Figure 26 shows the titration of the antimicrobial synergy chessboard.

Each square represents a tube. Increasing concentrations of antibiotic A on the horizontal axis and antibiotic concentrations on the vertical axis are disturbed by IR. In panel A, antibiotics show bonding effects; the isobologram on the right is a straight line. Panel B shows synergism where the isobologram is a concave curve. Panel C shows the opposite results with a curved curve.

Demonstration of an antibiotic susceptibility sample using the Kirby-Bauer method.

Table 22: Antibiotograms (antibioticograms) of isolates used for the study (zones in millimeters)

Antibiotic Area Size (mm) Organisms E.c Ps. MRSA Amikacin 20th g 22nd Ciprofloxacin - - 20th Kanamycin 14th - - Gentamicin 19th - - Tetracycline - - 27th

Nalidixic acid - - - Gefoperazone - 14th 23rd Ceftazidime 10th 16th - Chloramphenicol - - -

Keyword: no inhibition method.

Detection of ASAP clinical isolates by sensitivity diffusion

ASAP conc PPm Area size in millimeters Organisms E.coli Pseudomonas MRSA 32 16th 16th 13th 16th 15th 14th 11th 8th 11th 11th - 4 - - - 2 - - -

Keyword: no delay

See photos. Figure 27.

[607] Determination of Antibiotic Combination by Disk Zoom Assay.

Of the various antibiotic combinations tested for synergistic or coupling effects of isolates, inhibitory zones suggesting potential synergies were only observed with MRSA for the combination of Amikacin and Cefoperazone and Amikacin and Tetracycline (see Figure 28). The other two isolates, viz. In the case of E. coli and Pseudomonas, no inhibitory zones indicating synergistic combinations were observed (see Figures 29 and 30).

[608] Determination of Minimum Inhibitory Concentration (MIC) of Antibiotics.

MICs were determined for antibiotics which showed inhibitory zones suggesting potential synergies.

Table 23:

Amikacin MIC

Biomass: 125mcg / ml

Solvent: Nourishing broth

Culture: MRSA Key: +: Growth is not growth

The MIC of amikacin for MRSA element was found to be 0.8mcg / ml.

Cefoperazona MIC

Biomass: 100mcg / ml

Solvent: Nourishing broth

Culture: MRSA

Girl # Conc. (concentration) mcg / ml An increase 1 0.2 + 2 0.4 + 3 0.6 + 4 0.8 - 5 1. - 6th 2 - 7th 3 - 8th 4 - 9th 5 - 10th + ve + ii - ve -

Transcript: +: Growth is not growth

The Cefoperazone MIC for MRSA element was found to be 10mcg / ml ·

Silver / Water MIC

Raw materials: 20ppm silver / water solution

Girls no. Conc. (concentration) (approximately) An increase 1 5 + 2 10th - 3 15th - 4 20th - 5 25th - 6th 30th - 7th 35 - 8th 40 - 9th 45 - 10th 50 - 11th + ve + 12th - ve -

V

Solvent: Nourishing Broth Culture: MRSA

23a. table

Test tubes no. Conc. (concentration) (approximately) An increase 1 1 + 2 2 + 3 3 + 4 4 + 5 5 + 6th 6th + 7th 7th - 8th 8th - 9th 9th - 10th 10th - 11th + ve + 12th - ve -

Transcript: +: Increase

-: No growth

The Silver / Water MIC for MRSA element was found to be 8ppm.

Silver / Water MIC Biomass: 20 ppm Solvent: Nourishing Broth Culture: E.coli

Table 24

Test tubes no. Conc. (concentration) (approximately) An increase 1 1 + 2 2 + 3 3 - 4 4 - 5 5 - 6th 6th - 7th 7th - 8th 8th - 9th 9th - 10th + ve + π - ve -

Transcript: +: Growth is not growth

The silver / water MIC for the element E.coli was found to be 3ppm.

Silver / Water MIC

Biomass: 20 ppm silver / water

Solvent: Nourishing broth

Culture: Pseudomonas

Table 25

Test tubes no. Conc. (concentration) (approximately) An increase 1 1 + 2 2 + 3 3 - 4 4 - 5. 5 - 6th 6th - 7th 7th - 8th 8th - 9th 9th - 10th + ve + 11th - ve -

Transcript: +: Growth is not growth

The Silver / Water MIC for the Pseudomonas element was found to be 3ppm.

The synergistic action study was conducted using a chessboard type test.

I. Amikacin and Silver / Water Combination.

MIC of amikacin = 0.8mcg / mI.

Silver / water MIC = 8 ppm.

Culture: MRSA

1.6 - - - - - - - 0.8 - - - - - - - 0.4 + - - - - - - 0.2 - - - - - - 0.1 + 4 ~ - - - - - 0.05 ++ ++ - - - - - 0 + ve C ++ ++ ++ + - -

0.5 1 2 4 8 12

Vertical: Amikacin (mcg / ml) Horizontal: Silver / Water (ppm)

Transcript: +: Growth is not growth

The MRSA of the present invention was found to have a synergistic concentration of 0.05 mcg / ml for amikacin and silver for water.

Calculation of the FIC index:

Amikacin FIC = MIC for amikacin combination yes Amikacin alone MIC = 0.05 / 0.8 = 0.0625

ASAP parameter FIC = MIC for Silver / Water in combination Silver / Water alone MIC = 1/8 = 0.125

FIC Index = Amikacina FIC + Silver / Water FIC = 0.0625 + 0.125 = 0.1875.

The FIC index indicates the synergy between Amikacin and Silver / Water. [609] II. Cefoperazone and Silver / Water combination.

Cefoperazone MIC = 10mcg / ml.

Silver / Water MIC = 8ppm.

Culture: MRSA

15th - - - - - - 10th - - - - - - - 5 + - - - - - - 2.5 ++ - - - - - - 1.25 ++ - - - - - - 0.625 ++ ++ - - - - - 0 + ve c ++ +

0.5 1 2 4 8 12

Vertical: Cefoperazone (mcg / ml)

Horizontal: Silver / Water (ppm)

Transcript: +: Increase

-: No growth

MRSA was found to have a synergistic concentration of 0.625mcg / ml for Cefoperazone and ppm for Silver / Uden.

FiC Index Calculation:

Cefoperazone FIC = MIC for Cefoperazone in combination Cefoperazone MIC alone. = 0.625 / 10 = 0.0625

ASAP parameter FIC = MIC for Silver / Uden

Only silver / water MIC = 1/8 = 0.125

FIC Index = Amikacin FIC + Silver / Water FIC = 0.0625 + 0.125 = 0.1875.

The FIC index indicates the synergy between Cefoperazone and Silver / Water. [610] III. Combination of cefoperazone and amikacin.

Cefoperazone MIC = 10mcg / ml.

MIC of Amikacin = 8 ppm.

Culture: MRSA

15th - - - - - - - 10th - - - - - - - 5 + - - - - - - 2.5 ++ - - - - - - 1.25 ++ ++ ++ ++ - - - 0.62 5 ++ ++ ++ ++ + - - 0 + v e C ++ ++ ++

0.0 0, 0, 0, 0, 1, 5 1 2 4 8 6

Vertical: Cefoperazone (mcg / ml)

Horizontal: Amikacin (mcg / ml)

Transcript: +: Increase

-: No growth

Cefoperazone was found to have a binding concentration of 1.25 and amikacin 0.4. Calculation of the FIC index:

Cefoperazone FIC = MIC for Cefoperazone in combination Cefoperazone MIC alone.

= 0.125 / 10 = 0.125

Amikacin FIC - MIC for Amikacin in combination Only for MIC of Amikacin alone.

= 0.4 / 0.8 = 0.5

FIC index == Amikacma FIC + Cefoperazona FIC = 0.125 + 0.5 = 0.625.

The FIC index indicates sub-compositions between Cefoperazone and Silver / Water.

[611] DISCUSSION [612] In this study of three clinical isolates collected from P.D. In the example of a Hindu hospital in Mumbai, India, gram-negative isolates showed resistance to older antibiotics such as ampicillin, tetracycline, kanamycin and older carbostyrils such as nalidixic acid as well as third generation cephalosporin, ceftamizine and cefoperazone. The clinical isolates of Pseudomonas used in the study also proved to be resistant to the newest types of ciprofloxacin as well as the semi-synthetic aminoglycoside, amikacin. MRSA Gram-positive isolates also showed resistance to older antibiotics as well as to third-generation cephalosporins such as ceftazidime.

[613] A study of their sensitivity to the silver / water compositions of the present invention showed that Gram-negative isolates were sensitive to about 3ppm Silver / water solutions, whereas MRSA isolates were found to contain 8ppm Silver / water solutions determined using diffusion and macro-dissolution broth method.

[614] The interaction of these two antibiotics with isolates was determined using a disk diffusion method which showed synergy between cefoperazone and amikacin against MRSA. A chessboard type test was carried out to confirm this. No impurity or synergy was found between these antibiotics in the Gram-negative isolates during the disk diffusion test.

[615] A checkerboard type test was performed and the two antibiotics were found to have a FIC index of 0.625, thus indicating impurities rather than synergies with the combination of amikacin and cefoperazone.

[616] A checkerboard type test was also performed to investigate the combination of Silver / Water solution with amikacin as well as cefoperazone. The results obtained showed that, in the presence of the inventive silver / water compositions, the effective antibiotic concentration was reduced by about four units. The FIC index of these combinations was found to be 0.1875 in each case, indicating a synergy between the silver / water and amikacin and the silver / water and cefoperazone combinations.

[617] The results of the study indicate that in the above clinical MDR isolates the dose of antibiotics could be significantly reduced in the presence of silver / water, which is not the case with antibiotic combinations.

[618] These results demonstrate that the inventive silver / water combinations will play an important role in the therapy of antibiotic combinations, especially when dealing with many drug-resistant cases.

Table 26

1. Nourishing broth:

Peptone 10.0gr

Sodium chloride 5.0gr

Meat extract 3,0gr

Dextrose 5,0gr

Phenol red (indicator) 0.001%

Distilled water 900ml

2. Nourishing Agar

Peptone 10.0gr

Sodium chloride 5.0gr

Meat extract 3,0gr

Distilled water 900ml

Agar 2.0% pH

7.2

3. 'Muller and Hinton Agar Hydrolyzate of casein acid Beef starch Potato starch Agar

Distilled water PH

29.0gr

10.0gr

2.5gr

1.2% 100ml

7.6

Table 27

Interpretation of Area Diameter (NCCLS Document, 1988)

Antibiotic Drive Diameter of zone in millimeters Conc (concentration) (mcg) Resistant Average Sensitive Amikacin (AK) 30th <14 15-16 > 17 Ciprofolxacin (RC) 5 <15 16-20 > 21 Kanamycin (KA) 30th <13 14-17 > 18 Gentamicin (GM) 10th <12 13-14 > 15 Tetracycline (TE) 30th <14 15-18 > 19 Nalidixic acid (NA) 30th <14 14-18 > 19 Cefoperazone (CP) 75 <15 16-20 > 21 Ceftazidime (FG) 30th <14 15-17 > 18 Chloramphenicol (CH) 30th <12 13-17 > 18

[619] COMBINATION OF GENTAMICINE AND SILVER / AQUEOUS COMPOSITIONS AS A WINES ANTISEPTIC POWDER [620] INTRODUCTION [621] Wound antiseptic powders are a preparation technology for treating external (surface) bacterial infections of the skin, or for wound healing. .

[622] Wound powders are generally broad-spectrum antibiotics / antiseptics. The use of such powders does not preclude the concomitant use of antibiotic therapy where appropriate.

[623] Most of the wound treatment products on the market today are based on Povidone Iodine. Povidone iodine is highly cytotoxic in open wounds and has been specifically contraindicated in diabetic wounds. In addition, the iodine is sublimated and re-applied every 6-8 hours.

[624] Another potential application is in the veterinary field. Pets often suffer cuts, abrasions, and various wounds resulting from scratching to eliminate parasites and from collisions with other animals. Gentle but broad spectrum antimicrobials would be very useful in this case.

[625] It was decided to develop a wound antiseptic powder consisting of a slow release formulation and a combination of gentamicin and silver nanoparticles developed within the scope of the present invention. A talc-like preparation consisting of 200 ppm silver nanoparticles and about 100 ppm Gentamicin is referred to herein as SILDUST.

[626] RESULTS.

[627] SILDUST SENSITIVITY [628] Purpose: To determine the susceptibility of SILDUST and its components to microorganisms.

[629] Procedure:

[630] Equipment Required:

• Incubator, Laminar Flow [631] Materials Required:

• Sterile nutrient agar plates, sterile cotton swabs, micropipette (capacity 100μΙ - 1000 μΙ), 16-hour-old cultures (estimated density 10 CFU / ml), Escherichia coli (MDR), Pseudomonas aeruginosa (MDR), methicillin-resistant Staphylococcus aureus.

[632] Method:

• Apply 0.1 ml cultures to the surface of the nutrient agar using sterile cotton swabs. Leave in this position for 15 minutes.

• After 15 minutes, aseptically create wells on the agar surface using a 10 mm cork drill.

• Place 10 mg SILDUST (200 ppm Silver Talc + 100 ppm Gentamicin) in one well.

• Place 100 μΙ of lOOppm Gentamicin in the second well. Also add 200 ppm Silver Talc + 100 μΙ Distilled Water.

• Plates should be incubated at 37 ° C for 24 hours.

• Measure the zone of inhibition in millimeters using a HiMedia zone reader.

[633] Results: As shown in Table 28 and Chart 38.

Table 28: LABELS Sensitivity of ICT Culture Inhibition zone 100 ppm Gentamicin 200 ppm ASAP talc BRIDGING *

Escherichia coli 24mm 17mm 26 mm (MDR)

[634] Conclusion: Synergistic activity has been observed for SILDUST (containing 200 ppm silver talc and 100 ppm gentamicin).

[635] Transcript:

SILDUST 1 - 200 ppm silver talc + 50 ppm Gentamicin SILDUST 2 - 200 ppm silver talc + 100 ppm Gentamicin [636] SILDUST ANTIBACTERIAL ACTIVITY [637] Purpose: To determine the time of killing of SILDUST against microorganisms (time that SILDUST kills microorganisms).

[638] Procedure:

[639] Necessary equipment:

• Incubator, Laminar flow, scales.

[640] Required Materials:

• Sterile phenol red dextrose broth, 16 hours old cultures of these strains (approximately 10 ⁸ CFU / ml), Escherichia coli (MDR), Pseudomonas aeruginosa (MDR), methicillin-resistant Staphylococcus aureus.

[641] Method:

• Prepare a 5 ml aliquot of 2g SILDUST in a sterile test tube.

• Inject 0.1 ml of culture in the above solution. Stir well. At a time interval of 0, 5, 10 to 50 minutes, apply a loop of test sample to 5 ml of sterile phenol red dextrose broth. Stir well.

• Incubate at about 37 ° C for about 24 hours.

• Observe the increase.

• For negative control, the non-injected SILDUST loop was retained in 5 ml of sterile phenol red dextrose broth and incubated at approximately 37 ° C for approximately 24 hours.

For positive control, the test culture loop was injected with 5 ml of sterile phenol red dextrose broth and incubated at about 37 ° C for about 24 hours.

[642] Results: See Tables 29, 30 and 31.

Table 29 Escherichia coli ( MDR) At a time interval (minutes) 100 ppm Gentamicin 200 ppm ASAP Talc BRIDGING * Vocadine * 0 + + + - 5 + + + - 10th + + + - 15th + + + - 20th 4- + + - 25th + + + - 30th + + + - 35 + + 4 · - 40 4- + + - 45 + + + - 50 + + - - Positive control + + + + Negative control -

SILDUST * -> 200 ppm ASAP Talc + 100 ppm Gentamicin Vocadine * - »200 ppm available iodine

Transcript:

+ - ^ Growth -> No Growth [643] Conclusion: This mixture shows synergistic action. The VOCADINE tube (described below, at the end of the example) became brown in color a few seconds after adding the powder to the agent due to iodine release. Although VOCADIN shows faster killing, such a high level of cytotoxicity is not desirable for wound healing.

[644] Tab a Pseudomonas aeruginosa (MDR) At a time interval (minutes) 100 ppm Gentamicin 200 ppm ASAP Talc BRIDGING * Vocadine * 0 + + + - 5 + + + - 10th + + + -

15th + + + - 20th + + + - 25th + + + - 30th + + + - 35 + + + - 40 + + - - 45 + + - - 50 - - - - Positive control + + + + Negative control - - - -

SILDUST * -> 200 ppm ASAP Talc + 100 ppm Gentamicin

VOCADINE * -> 200 ppm available iodine

Transcript:

-l—> Growth -> No Growth [645] Conclusion: This combination shows synergistic action.

[646] Tab MRSA At a time interval (minutes) 100 ppm Gentamicin 200 ppm ASAP Talc BRIDGING * Vocadine * 0 + + + - 5 + + - 10th + Ί ^- - - 15th - + - - 20th - + - - 25th - + - - 30th - - - - 35 - - - - 40 - - - - 45 - - - - 50 - - - - Positive control + + + + Negative control - - -

SILDUST * - »· 200 ppm ASAP Talc + 100 ppm Gentamicin

VOCADINE * -> 200 ppm available iodine

Transcript:

+ -> Growth - - »No Growth [647] Conclusion: This combination shows synergistic action.

[648] ANTIBACTERIAL PERFORMANCE OF SILDUST [649] Purpose: To determine the susceptibility of a bacterial virus to SILDUST.

[650] Principle: An appropriate solution must be obtained to prevent a false positive test due to the fact that SILDUST kills the host organism.

[651] Procedure:

[652] Principle:

• T-pair bacteriophages and Escherichia coli host are used as a detection system. The silver concentration in SILDUST should be neutralized by dilution to avoid killing the host. Experimental aliquots were prepared in this manner;

1. Test -Phage + WARM

2. Control - Phage + Saline [653] Equipment Required:

• Scales, laminar air flow device, incubator.

[654] Materials Required:

• Petri plates, marker, spatula, micropipette.

[655] Method:

• Prepare a 5 ml aliquot containing approximately 1 gr of SILDUST (showing no bactericidal effect) and saline in separate sterile test tubes.

• Add approximately 0.1 ml of phage lysate to each (approximately ¹⁰ IU of phage particles per ml).

• Mix well in a vortex mixer and incubate at about 37 ° C.

• At t = 0, 1 and at intervals thereafter, withdraw 0.5 ml aliquot and dilute in experimental SILDUST solution, which has no bactericidal effect.

• Spray this solution on a converging freshly prepared host screen. This needs to be done for both the test and the controls.

• Incubate the plate at approximately 37 ° C for approximately 24 hours.

• Mix 0.1 ml of this solution with 0.5 ml of incremental host and incubate at 37 ° C for approximately 15 minutes.

• Add 7 ml of weak melted agar to it.

• Stir well and coat on St. nutritious agar plates.

• Incubate the plate at approximately 37 ° C for approximately 24 hours.

• Check the screen for inflammation spots and number the inflammation spots on the coating.

[656] Results: See Table 32

Table 32 - BRIDGE Experiment

Solutions Result 10 ' ¹ + lu ² - 10 ' ^J - And ⁴ -

Transcript: + -> Presence of active phage particles

- - + No active phage particles present [657] SILDUST - VIRUS KILLING OPERATION [658] Purpose: To detect SILDUST virus killing (antiviral) activity by a bacteriophage detection system.

[659] Procedure: As in "HEAT ANTIBACTERIAL OPERATION, Part 2" [660] Results: As shown in Tables 33 and 34.

[661] Table 33 SILDUST killing time

At a time intervals (hours) Saline solution BRIDGING * 0 + + 1 + + 2 + ' - 3 + -

SILDUST * - * 200 ppm ASAP talc + 100 ppm Gentamicina Key: + -> Active phage particles not active phage particles [662] Table 34

At a time Saline solution BRIDGING * intervals (pfu / ml) (pfu / ml)

(hours) 0 TNTC 1.15 X 10 ⁵ 1 TNTC 1.0 X 10 ⁴ 2 TNTC 3.0 Χ10 ³ 3 TNTC Nil

BRIDGE * -> 200 ppm ASAP talc + 100 ppm Gentamic

Transcript: TNTC -> Too Numerous To Count pfu / ml - Infectious Phage Particle Titer [663] Conclusion: SILDUST was found to exhibit bactericidal activity against the host culture at 10 ' ² solution. The same SILDUST solution was tested for antiviral activity and found to be effective. It was also found that the inflammatory plaque-forming units decrease from 10 ⁵ to zero within 3 hours, demonstrating that the SILDUST element can also act against animal viruses.

The following compositions were used in the above experiments:

[664] Environmental Composition Nutrient Agar:

Peptons 10.0 gr

Sodium chloride 5.0 gr

Meat extract 3.0 gr

Distilled water 900 ml

Agar 2.5 gr pH 7.2 ± 0.2

Phenol red dextrose broth

Proteozoal peptone 10.00 g / It

Beef extract 1,00 g / lt

Sodium chloride 5.0 g / lt

Dextrose 5.0 g / lt

Phenol red 0.018 g / l pH 7.4 ± 0.2

Weak Agar

Agar 1.0%

Saline solution

Sodium chloride 0.9%

VOCADIN

Production License No. AD / 200-A

Serial No .: WNR5008

Preparation Date: March 2005

Expires: March 2008

Active ingredients Povidone iodine IP 5% w / w

Navketan Research and Lab. Ltd.

[665] SILVER / WATER IMPURITIES FOR 10% POVIDON IODINE SOLUTION [666] Another impurity that successfully works in the silver / water compositions of this invention is Povidone Iodine. Iodine is a well-known prophylactic drug for the treatment of a wide range of pathogens. Various concentrations of iodine are commercially available, but the most commonly used and recommended concentration is 10%. In this preferred embodiment of the present invention, a synergistic combination of 25-50% is a silver / water composition replacing 10% iodine solution. Although some reactions are possible between the silver / water composition and iodine, the results of the experiment indicate that the synergistic combination of silver / water and Povidone iodine may act as a disinfectant (such as an ointment) and / or as a preventive agent against wounds, burns and burns. scratches, etc. in case.

[667] The synergistic action of 32ppm silver / water compositions at various percentages on Povidone iodine (PI) against many bacteria has been studied with particular care. This results in test methods and results. From the obtained results it can be concluded that there is a synergistic interaction between the two materials. This synergism can be used to obtain a great local disinfectant.

[668] The following statements include what has been specifically illustrated and described above, which is conceptually equivalent, what can be obviously replaced, and what essentially incorporates the basic idea of the present invention. Skilled and skilled professionals will appreciate that multiple adaptations and modifications of this just described embodiment can be configured without departing from the scope of the invention. The illustrated embodiment is provided for illustrative purposes only and should not be construed as a limitation of the invention. Therefore, it is to be understood that, within the scope of the appended claims, the invention may be practically practiced otherwise than as set forth herein.

Claims (31)

A composition of silver in water containing a total concentration of silver between 5 and 40 particles per million, said silver in the form of silver nanoparticles internally as elemental silver and as a surface with at least one silver oxide, wherein most silver particles have a maximum diameter less than 0.015 micrometres most of the colloidal silver particles have a minimum diameter greater than 0.005 micrometers and in which the composition exhibits antimicrobial properties. A composition according to claim 1, further comprising hydrogen peroxide. The composition of claim 2, wherein the concentration of hydrogen peroxide is between 1% w / v and about 3.0% w / v. The composition of claim 1, further comprising EDTA. The composition of claim 4, wherein said EDTA comprises disodium EDTA. The composition of claim 1, wherein the composition comprises a hydrogel formed by dissolving a hydrophilic polymer in a silver compound in water. The composition of claim 6, formed as an amorphous gel. The composition of claim 6, which is in the form of a solid gel sheet. The composition of claim 8, wherein the hydrophilic polymer is selected from the group consisting of gelatin, carbohydrate polymers, and copolymers of acrylic acid. The composition of claim 9, wherein the carbohydrate polymer comprises at least one polymer selected from the group consisting of cellulose derivatives, alginate, carrageenan, and plant resin. Use of an aliquot of the composition according to claim 1, for use as a step in the treatment of a disease selected from the group consisting of malaria, fungal infections of the skin, bacterial skin infections, vaginal infections, urinary tract infections, tonsillitis, inflammation of the pelvic organs, for the treatment of pharyngitis, gonorrhea, conjunctivitis, otitis and respiratory tract diseases and nasal infections in a person suffering from the disease. 12. Use of silver EDTA as a step in the treatment of a disease selected from the group consisting of malaria, fungal infections of the skin, bacterial skin infections, vaginal infections, urinary tract infections, tonsillitis, pelvic inflammation, pharyngitis, gonorrhea, conjunctivitis, otitis. and respiratory diseases and nasal infections, treatment. 13. A microbe selected from the group consisting of Baditus anthrads, Badllus t subtilis, Candida albicans, Mycobacteria bovis, Mycobacteria tuberculosis, A method of eradication of Pseudomonas aeruginosa, Salmonella choleraesius, Staphylococcus aureus, Trichomonas vaginalis and Yersinia pests, which comprises exposing said microbes to silver EDTA. The method of claim 13, wherein said exposure comprises oral ingestion of silver EDTA. 15. A microbe selected from the group consisting of Baditus anthrads, Bacillus subtilis, Candida albicans, Mycobacteria bovis, Mycobacteria tuberculosis, Pseudomonas aeruginosa, Salmonella choleraesius, Staphylococcus aureus, Trichomonas vaginalis, and Yersinia metis, exposing at least one composition selected from the group consisting of silver EDTA, silver EDDS, silver curcuminates, silver berberine and silver tetracycline. 16. A method of delivering at least one metal to a biological organism, comprising the addition of at least one metal selected from the group consisting of silver, copper, zinc, platinum, titanium and mixtures and alloys thereof for at least one clathrate to form a metal / clathrate structure, and subjecting said biological organism to said metal / clathrate structure. The method of claim 16, wherein said clathrate comprises at least one kaolinite. The method of claim 16, wherein said clathrate comprises at least one zeolite. The method of claim 16, wherein said at least one metal contains silver. 20. Method of use of AgEDTA for the addition of at least one livestock feed and livestock water for the prophylactic feeding of livestock. 21. A method of using AgEDTA, when added to anything that a human or animal receives, for the prophylactic treatment of said human or animal. The method of claim 21, wherein said AgEDTA is added in an amount sufficient to prevent infections. The method of claim 21, wherein said AgEDTA is added to the compound of claim 1 in an amount of less than 20 parts per minute. 24. A method of using AgEDTA in an amount sufficient to ameliorate an infection to treat said human and animal infections. 25. A method of using at least one element selected from the group consisting of AgEDTA, silver EDDS, silver curcuminates, silver berberine and silver tetracycline for the treatment of human or animal infections. 26. A method of using a paste or gel comprising at least one element selected from the group consisting of AgEDTA, silver EDDS, silver curcuminates, berberine silver and tetracycline silver, and contacting the human or animal skin surface with said human or animal skin. skin surface treatment. A gel or paste product comprising at least one element of the group consisting of AgEDTA, silver EDDS, silver curcuminates, berberine silver and tetracycline silver. A method of improving the dose of an antibiotic comprising adding at least one substance selected from the group consisting of EDTA and AgEDTA to said dose of the antibiotic. The method of claim 28, wherein AgEDTA is added to said dose of antibiotic. The method of claim 11, further comprising adding a dose of a selected antibiotic, said dose of said antibiotic being based on antibiotics having at least some known efficacy against said disease. The composition of claim 1, further comprising at least one substance selected from the group consisting of AgEDTA, silver EDDS, silver curcumin, berberine silver and silver tetracycline.