KR100817029B1 - Development of gastric protected slilver nanoparticle and their manufacturing method - Google Patents

Abstract

A method of manufacturing silver nanoparticles of a protective coating stable without decomposition in the stomach is provided to give silver nanoparticles having effects of causing no adverse effects when fed to a pig and poultry, increasing productivity and reducing the generation of odors in livestock facilities and capable of being well absorbed in the intestine and used as a feed additive. Silver nanoparticles stable without decomposition in the stomach are prepared by the steps of: immobilizing a silver nanoparticle colloidal solution and nutrients to a water insoluble carrier; mixing the fixed silver nanoparticle colloidal solution and a coating agent and stirring; and encapsulating the mixture. The silver nanoparticle colloidal solution has a particle size of 1 to 15nm and contains one or more polymeric stabilizers selected from the group consisting of polyvinylpyrrolidone, (1-vinyl pyrrolidone)-acrylic acid copolymer, polyoxyethylene stearate and (1-vinylpyrrolidone)-vinyl acetic acid copolymer. The carrier is selected from the group consisting of silica, cellulose, starch, zeolite, calcium carbonate, wheat red dog and corn flour. The coating agent is selected from the group consisting of a monovalent soap, divalent soap, hydrogenated vegetable oil and the like.

Description

Stable protective coating on silver nano and its manufacturing method {Development of gastric protected slilver nanoparticle and their manufacturing method}

The present invention provides a method for producing silver nano-coated protective coating through a coating process so that silver nanoparticle colloid having a particle diameter of 1 to 15 nm can be absorbed by moving to the lower digestive system stably without decomposing in the stomach and to feed pigs and poultry. To develop feed additives.

Silver basically has excellent features such as sterilization, deodorization and antimicrobial effect against more than 650 pathogenic bacteria.Silver nano silver, which is broken by nano-ization, has stronger antibacterial, sterilization and deodorization function than the original silver. Have In addition, while having a sterilizing power tens of times stronger than chlorine series, it has harmless characteristics in the body. Patent Publication No. 2005-0097305 discloses a description of the antimicrobial activity of silver and Patent No. 517019 discloses a food waste dryer in which a partition plate that is coated with silver nano-deodorizing function is formed. Patent Publication Nos. 2003-0082064 and 2003-0082065 disclose that silver nanoparticles can be used as antibacterial agents, fungicides and deodorants. However, although the above-described prior art has limitations of use, when silver nanoparticles are synthesized or mixed with other substances, silver nanoparticles are easily destabilized, and even when fed into the body, they lose their intrinsic properties due to the instability of silver in the stomach. There is a big difficulty in commercialization.

In order to solve the problems of the prior art, the inventors have made it possible to maintain the intrinsic properties of silver even if the silver nanoparticles are synthesized or mixed with other materials, and the development of protective coating silver nanopore of pig and poultry type for livestock applications. Was completed.

The present invention is to provide a coating technology that can be absorbed after stable movement in the intestine without decomposition in the stomach of livestock.

It is also an object of the present invention to provide a method of applying pig and poultry feed additives using the composition.

The present invention relates to a method for producing silver nano-coated silver nanoparticle colloid so that it can be stably moved and absorbed into the lower digestive tract without being broken down in the stomach and feed additives for feeding pigs and poultry.

The silver nanoparticle colloidal solution in the composition of the present invention is a polyvinylpyrrolidone, (1-vinyl pyrrolidone) -acrylic acid copolymer, polyoxyethylene stearate and (1-vinylpyrrolidone) -vinyl acetic acid copolymer A composition comprising one or more polymer stabilizers selected from the group consisting of. The polymer stabilizer may be included in the range of 0.5 to 5% by weight based on the total weight of the composition. These polymer stabilizers are safe in animals and in vitro, but allow the silver nanoparticles to be evenly dispersed in the colloidal solution. Therefore, it is possible to evenly deliver the silver nanoparticles contained in the composition of the present invention in and out of the animal.

The silver nanoparticle colloid used in the composition of the present invention may be prepared according to a method for preparing silver nanoparticle colloid known in the art. For example, WO 02/087749 discloses a method for preparing a colloidal solution.

Silver nano-manufacturing method according to the present invention is as follows.

First, the silver nano colloidal solution of the above-mentioned liquid is immobilized on a carrier to make the silver nano component immobilized so that the silver nano component can be protected in the stomach. Wherein the silver nanoparticle colloidal solution is a silver salt or silver oxide salt with polyvinylpyrrolidone, (1-vinylpyrrolidone) -acrylic acid copolymer, polyoxyethylene stearate and (1-vinylpyrrolidone) -vinyl acetate copolymer In the group consisting of one or two or more mixtures are preferable, and the carrier is preferably a water-insoluble carrier selected from the group consisting of powder, jade, silica, cellulose, starch, zeolite and calcium carbonate, and more preferably silica. It is preferable.

Next, the encapsulation coating agent was encapsulated in the silver nano component immobilized on the prepared carrier to prepare encapsulated silver nanoparticles in the form of granules or needles. The encapsulation was carried out in an extruder such as an extruder or a plotter after mixing and stirring in a mixer such as a super mixer, a ribbon mixer, a general stirrer, a homo mixer, and a V-type mixer. Herein, the weight ratio of the immobilized silver nano and the binder coating agent is preferably 1: 1 to 99, and the porous plate size of the extruder is preferably 0.5 to 2.5 mm. In addition, the binding coating agent preferably contains one or two or more monohydric soap, dihydric soap and fatty acid having a melting point of 40 ℃ or more or extremely hardened vegetable oil. The monovalent soap is preferably sodium soap or potassium soap, and the dihydric soap is preferably selected from the group consisting of soaps composed of calcium, magnesium and zinc. The fatty acid having a melting point of 40 ° C. or higher is preferably selected from the group consisting of palmitic acid and stearic acid, and the extremely hardened vegetable oil is preferably selected from the group consisting of hardened palm oil and hardened palm stearin having a melting point of 57 ° C. or higher. Do. The encapsulated silver nano is preferably a binding coating agent content of 50 to 99%.

The encapsulated silver nanoparticles prepared in accordance with the present invention have the advantage of exhibiting uniform and uniform appearance. In addition, the silver nano component is bypassed to the intestine without decomposing or losing from the stomach than when directly fed uncoated silver nano, thereby increasing utilization efficiency. In addition, the production cost of the product is lowered due to the advantages of low manufacturing cost and low site and equipment cost. Because of this, feeding silver nano in an unprotected form is less effective and wasteful. Therefore, the development of protective silver nano to use in livestock specification to increase the nutrient utilization efficiency of pigs and poultry to increase productivity, and to develop various functional feed additives.

Hereinafter, the present invention will be described in more detail with reference to Examples, but the protection scope of the present invention is not limited to the following Examples.

Example  1: encapsulated Silver nano  Manufacturing method

Experimental Example 1

 0.5kg of silver nanoparticle colloidal solution with 10,000ppm silver content is mixed with 0.5kg of silica, which is immobilized carrier, and then immobilized.Then, 70kg of calcium soap and 29kg of hardened palm stearin are added to the super mixer by 1.2mm by extruder. Injection was carried out through the trial to produce encapsulated and granular encapsulated protective silver nano. The ratio of the binder coating agent of the protective silver nano prepared as described above is 99%.

Experimental Example 2

1kg of silver nanoparticle colloidal solution with 10,000ppm silver content is mixed with 1kg of silica as immobilization carrier, and then immobilized, and then mixed with 69kg of calcium soap and 29kg of hardened palm stearin in a super mixer. Injection was carried out to prepare encapsulated and granular encapsulated protective silver nanoparticles. The ratio of the binder coating agent of the protective silver nano prepared as described above is 98%.

Experimental Example 3

5kg of silver nanoparticle colloidal solution with 10,000ppm silver content is mixed with 5kg of silica, which is immobilized carrier, and then immobilized, and then mixed with 63kg of calcium soap and 27kg of cured palm stearin in a super mixer. Injection was carried out to prepare encapsulated and granular encapsulated protective silver nanoparticles. The ratio of the binder coating agent of the protective silver nanoparticles prepared as described above is 90%.

Experimental Example 4

10 kg of silver nanoparticle colloidal solution with 10,000 ppm of silver is mixed with 10 kg of silica, which is immobilized carrier, and then immobilized. Injection was carried out to prepare encapsulated and granular encapsulated protective silver nanoparticles. The ratio of the binder coating agent of the prepared protective silver nano is 80%.

Experimental Example 5

20 kg of silver nanoparticle colloidal solution with 10,000 ppm of silver is mixed with 20 kg of silica, which is immobilized carrier, and then immobilized. Injection was carried out to prepare encapsulated and granular encapsulated protective silver nano. The ratio of the binder coating agent of the protective silver nano prepared as described above is 60%.

Experimental Example 6

1kg of silver nanoparticle colloidal solution with 10,000ppm silver content is mixed and immobilized in 9kg of powder, which is immobilized carrier.Then, 63kg of calcium soap and 27kg of hardened palm stearin are mixed in a super mixer. Injection was carried out to prepare encapsulated and granular encapsulated protective silver nanoparticles. The ratio of the binder coating agent of the protective silver nano prepared as described above is 90%.

Example  2: encapsulated Silver nano in in vitro Loss rate  Measure

In vitro loss ratio of the encapsulated silver nanoparticles prepared in Experimental Examples 1 to 6 of Example 1 was measured by the following method, and the results are shown in Table 1.

In order to measure the stability of the encapsulated silver nanoparticles prepared in Experimental Examples 1 to 6, the prepared protective silver nanoparticles were placed in a test tube, hydrochloric acid and distilled water were added to prepare a 5% aqueous solution adjusted to pH 3, and then the aqueous solution was prepared. The mixture was stirred at 40 ° C. ± 0.1 ° C. for 1 hour using a JEIO TECH MC-11 multistirrer water bath.

The aqueous solution was filtered using Whatman paper No. 3 to take only solids, and the solids were dried at 45 ° C. ± 0.1 ° C. for at least 12 hours to completely remove residual moisture.

The dried sample was cooled to room temperature in a SANPLA DRY KEEPER and the cooled sample was then measured for silver content through an atomic absorption spectrometer (AAS). In addition, as a control, a comparative experiment was conducted in the same manner using a product immobilized on silica without coating the silver nano colloidal solution. Here, all assays were repeated three times, and the background experiments were performed in parallel.

In vitro loss rate for encapsulated silver nano Silver content (ppm) in vitro loss rate (%) Experimental Example One 50.2 10.5 2 100.1 12.6 3 500.5 17.8 4 998.5 25.3 5 2003.2 46.5 6 100.5 13.8 Control 100.5 95.9

Example  3: minimum growth inhibitory concentration against major pathogenic microorganisms ( MIC Confirmation of)

Minimum growth inhibition concentrations were determined based on known methods (Xu Hong? Xi and Lee Song F. Rhytother res, 18: 647-651, 2004). First, 5 outdoor strains and 6 standard strains were measured according to the broth dilution method.

As the silver nanoparticle colloidal solution used for the MIC measurement, the purified silver nanoparticle colloidal solution obtained by the above procedure was used. The silver nanoparticle colloidal solution was diluted in sterile distilled water to a final concentration of 1,000 ug / ml, and then filtered using a 0.2 μm filter (Millipore, USA). Next, 11-fold dilution (up to 211-fold) multi-step dilution with Mueller hinton broth (Difco, USA) was performed 11 times, and 100 μL was dispensed into each well of a 96 well plate (Corning, USA). Next, the strains enriched in tryptic soy broth (Difco, USA) were adjusted to MacFarland 0.5 turbidity (1.5x10 ⁸ CFU / ml), and then inoculated into each well by 10 µl and incubated for 24 hours. It was. Absorbance was measured at 600 nm with an ELISA reader (Tecan, Australia) to determine the minimum inhibitory concentration.

The measurement result of the minimum inhibitory concentration is shown in Table 2 below.

Minimum Inhibitory Concentration of Silver Nanoparticle Colloids for Major Pathogenic Microorganisms Strain Minimum Inhibition Concentration (㎍ / ㎖) Outdoor strain ^a Staphylococcus aureus 15.6 Staphylococcus epidermis 7.8 Streptococcus uberis 7.8 Esherichia coli 15.6 Klebsiella pneumoniae 7.8 Standard strain ^b Staphylococcus aureus  305 15.6 Staphylococcus epidermis ATCC 12228 7.8 Streptococcus uberis ATCC 27958 3.9 Streptococcus agalactiae ATCC 13813 3.9 Esherichia coli O55 15.6 Bacillus subtilis ATCC 1024 1.9

Field strain ^a : Livestock Research Institute isolated strain, standard strain ^b : National Veterinary Research and Quarantine Service

As shown in Table 2, the minimum inhibitory concentration of the silver nanoparticle colloid varies depending on the strain, but Staphylococcus aureus and Escherichia Coli showed high minimum inhibitory concentration. Silver nanoparticle colloids have been shown to have excellent antimicrobial activity against most pathogenic microorganisms.

Example  4: pig Protective coating silver nano Stink of Reduction effect

In this embodiment, as a part of reducing the odor, which is a chronic disease of pig farms, the protective coating silver nano is fed to measure the concentration of the gas generated in the pig company to confirm the effect of reducing the odor. Gas concentration measurement was performed using a gas meter, pump set MODEL.GV-100S (Gastech Co., Ltd.), inhaling 100 ml of gas at a position 3 cm above the surface of the central ventilator position inside the pig house The amounts of ammonia and amine gas components were then read. Ammonia and amines were measured using a gas measuring bar (disposable glass material) of the gas measuring instrument and the pump set MODEL.GV-100S (Gastech Co., Japan) used for the measurement of the amount of gas.

The protective coating silver nano supplemented to the pigs in this experimental example was added to the general feed by 100 ppm concentration prepared in Example 2, 5g / head / day per day. The farm selection was a bad odor and farms with 2000 heads were selected. The farm was selected for five months from May 2006 to the end of September. The results are shown in Table 3 below.

Determination of odorous gas concentration in pig houses Type of gas May to June July to August September Average ammonia Before pay 29.0 ppm 20.0 ppm 35.0 ppm 28.0 ppm After salary 14.0ppm 11.0 ppm 15.0 ppm 13.3 ppm Removal rate 51.7% 45.0% 57.1% 51.3% Amines Before pay 91.0ppm 74.0ppm 97.0ppm 87.3 ppm After salary 41.0ppm 38.0 ppm 39.0 ppm 39.3 ppm Removal rate 54.9% 48.7% 59.8% 54.5%

As shown in Table 3, after the protective coating silver nano, it was found that more than 50% of the ammonia and amine gas generated in the pig house can be removed.

Example  5: for pigs Protective coating silver nano  Effect of Salary on Growth of Pigs

In this embodiment, it was paid in the same manner as in Example 4, and was carried out at the Rural Livestock Research Institute for 40 piglets at weaning season. The results are shown in Tables 4 and 5 below.

Effect of protective coating for pigs on weight gain after silver nano supplementation Treatment history Disclosure weight (35 days old, kg) End weight (56 days old, kg) Weight gain (kg) Daily weight gain (kg) Remarks General feed group (20) 10.6 18.69 8.09 0.40 Protection coating silver nano 21 days salary Protective coating silver nano salary group (20) 10.9 21.7 10.8 0.54

Effect of protective coating on pigs on diarrhea after feeding nano Diarrhea Protective coating silver nano salary group (20 heads) General feed group (20) Number of occurrences 0 3 Doubtful head One 7

As shown in Table 4, the protective coating was confirmed that the increase in piglets weight gain when the silver was fed, and as shown in Table 5, the decrease in the frequency of diarrhea was investigated. The protective coating according to the present invention can be seen that the reduction of intestinal diarrhea and increase in weight increase by the reduction of intestinal pathogenic bacteria due to the intestinal retention of silver nano with excellent antimicrobial activity, improved digestibility.

Blood Change after Pig Protection Treatment history Glucose (85-150) GPT (31-58) GOT (32-84) White blood cell (11-24) Red blood cells (5-8) Platelets (325-715) General feed salary group (20) 121 38.30 43.24 23.80 9.01 636 Protective coating silver nano salary group (20) 145 (↑) 53.41 52.22 23.31 9.82 (↑) 774 (↑)

Blood changes used in this experiment were measured using an automated biochemical analyzer (Acro, USA) and hemocytometer (Hema VET-960, USA) within 1 hour after collecting blood from the jugular vein of pigs.

As shown in Table 6, the protective coating showed that the increase in feed intake and the increase of cell metabolism after nano supplementation resulted in an increase in the level of glucose, erythrocytes and platelets in the blood. It was confirmed that it does not act as a liver toxicity factor.

Example  6: poultry Protective coating silver nano  Odor of salary Reduction effect

In this embodiment, the protective coating silver nano to the broilers were fed to measure the concentration of the gas generated in the cage to determine the efficacy of reducing the odor. Gas measurement was performed in the same manner as in Example 4.

The protective coating silver nano fed to broilers in this Example was added to the general feed of 100 ppm concentration prepared in Example 1, 0.2g / allowance / day. The farms were selected for farms with severe odors and a breeding scale of 50,000, and were conducted for 33 days from July 2006 to early August. The results are shown in Table 7 below.

Determination of odorous gas concentration in cages (unit: ppm) division Ammonia Gas Concentration Amine gas concentration General feed salary group (12,000 number) 13 42 Protective coating silver nano salary group (12,000 number) 6 18

As shown in Table 7, after the protective coating silver nano, it was found that more than 50% of the ammonia and amine gas generated in the cage can be removed.

Example  7 poultry Protective coating silver nano  Effect on the growing season of broiler chickens

In this example, the same method as in Example 6 was used to carry out the same breeding farm.

Effect of protection coating for poultry on broilers after silver nano feeding division Early breeding Dead shooter Shipment Shipping weight (kg) Average weight (kg) Feed rate Protective coating silver nano salary group 12,000 305 11,695 19,032 1.62 1.64 General feed salary 12,000 427 11,573 18,130 1.56 1.71

As shown in Table 8, it can be seen that the mortality and average weight of the protective coating silver nano-supply group were significantly higher than those of the non-paying group. It can be seen that it increases the efficiency of the

Example  8 poultry Protective coating silver nano  Cholesterol Reduction Effect of Feeding

In the present embodiment was fed in the same manner as in Example 6, broilers were carried out in the same breeding farms, laying hens were carried out on a farm of 100,000 breeding scale.

Changes in Cholesterol after Feeding Silver Nanoprotective Coatings for Poultry (mg / 100g) division Broiler egg brisket Leg meat General feed salary 50.5 89.3 420.4 Protective coating silver nano salary 41.8 77.2 304.3

Cholesterol analysis used in this example was analyzed according to the Food Code (2006, General Ingredient Test Method). As shown in Table 9, the protective coating silver nano supplementation is involved in the body metabolism of broilers and laying hens to reduce the concentration of saturated fatty acid cholesterol and intramuscular and egg cholesterol by 20%.

Example  9: Protective coated Silver nano  Feed additives and liquids ( Untreated ) Silver nano  Measurement of stomach and intestinal concentrations after salary

In this example, the concentration of silver nano in the stomach and intestine was measured after feeding the liquid silver nano-protective coated pig and poultry silver nano-feed additive in pellet form. Feeding protective coating silver nano was added to the general feed at 100 ppm concentration prepared in Example 1, 5g (pig), 0.2g (chicken) / head per day. Feeding period was five days. After the experiment, 5 pigs and 10 broilers were collected and silver nano concentrations were measured for each organ. Silver concentrations were measured using inductively coupled plasma emission spectroscopy (ICP-AES). In summary, take an appropriate amount of sample, place it in a beaker, add nitric acid and glass spheres, and heat. This process removes obstructions and repeats until the solution is clear. Purified water is added to it, and the residue is dissolved and diluted to finish pretreatment. The pretreated sample was measured for the emission line intensity at 358.068 nm using ICP-AES, and since the intensity of the emission line was proportional to the concentration, the silver concentration was calculated by converting the concentration value through the calibration curve.

Intestinal silver nano concentration after protecting silver and liquid (non-treated) silver nano for pigs division Silver Nano Concentration (㎍ / kg) Stomach Duodenum Plant (jejunum) Ileum Cecum Colon Protective coating silver nano salary group ^A 1.27 7.54 12.88 15.72 10.77 5.94 Liquid silver nano salary group ^B 12.81 4.24 3.14 1.58 0.98 0.87 Loss Rate (B / A) - 43.8% 75.7% 89.9% 90.1% 85.4%

Intestinal silver nano concentration after protection silver and liquid (non-treated) silver nano for poultry division Silver Nano Concentration (㎍ / kg) Proventriculus Duodenum Plant (jejunum) Ileum Cecum Colon Protective coating silver nano salary group ^A 1.45 1.24 2.54 3.11 2.23 0.98 Liquid silver nano salary group ^B 3.21 2.21 1.14 0.87 0.74 0.55 Loss Rate (B / A) - - 55.2% 72.1% 64.9% 43.3%

As shown in Table 10, after digestion of silver-coated silver nano-protection for pigs, disintegration rarely occurred, and the intestinal retention began in the duodenum, indicating the highest concentration in ileum. In broilers of Table 11, the concentration of intestinal storage was higher in the plant than in the liquid silver nano feeding group, and the highest concentration of silver in the ileum was measured. This resulted in minimizing the decomposition of silver nanoparticles in the stomach compared to the existing liquid silver nanoparticles and confirmed that it can store large amounts of silver nanoparticles in the intestine.

The present invention is a silver nano-coated protective coating so that it can be safely absorbed after moving in the intestine, without being disintegrated in the stomach of the livestock, is an invention designed in the form of pellets so as to be easy to use as feed additives of pigs and poultry having the unique characteristics of silver nano.

When feeding pigs and poultry, there is no systemic side effect and excellent stability, and it can contribute to the establishment of environment-friendly livestock industry by increasing productivity and reducing the occurrence of odor in the barn. The development of feed additives using silver nano is the first of the present invention, and is expected to bring enormous economic benefits from domestic as well as foreign exports.

Claims (8)

In the method for producing a stable silver nanoparticles without decomposition in the stomach, (1) immobilizing the liquid silver nanoparticle colloid and nutrient on a water insoluble carrier; (2) mixing and stirring the binding coating agent in the silver nano and nutrients immobilized in step (1) in a mixer; And, (3) a stable protective coating silver nano production method of the above, characterized in that consisting of encapsulating the mixture prepared in step (2) via an extruder. The silver nanoparticle colloid of claim 1, wherein the silver nanoparticle colloid is a silver nanoparticle colloid solution having a particle size of 1 to 15 nm, and polyvinylpyrrolidone, (1-vinyl pyrrolidone) -acrylic acid copolymer, polyoxyethylene stearate. A method of producing protective nano-coatings as described above, comprising at least one polymer stabilizer selected from the group consisting of a rate and a (1-vinylpyrrolidone) -vinyl acetate copolymer. The method of claim 1, wherein the carrier is selected from the group consisting of silica, cellulose, starch, zeolite, calcium carbonate, powder, and jade. The method of claim 1, wherein the binding coating agent monovalent soap, divalent soap, vegetable hardened oil, extremely hardened fats and oils, hardened vegetable oil, hardened corn oil, hardened cotton seed oil, hardened peanut oil, hardened palm kernel oil, hardened palm oil, hardened palm The method of claim 1, wherein the protective coating is stable, characterized in that the one or two or more mixtures selected from the group consisting of fatty acids such as stearin oil, cured sunflower oil, cured vegetable oil, palmitic acid, stearic acid. The protective coating silver nano composition prepared by the method according to claim 1. The protective coating silver nano composition according to claim 5, which has an antibacterial and anti-odor effect on the livestock products. 6. The protective coating silver nano composition according to claim 5, wherein the frequency of diarrhea and the average weight of the livestock products are reduced compared to the general feed which does not include the protective coating silver nano composition. A method for producing livestock products, comprising feeding the silver nano composition of claim 5 to livestock.