KR102049673B1 - Process for Compositions of Disinfectants for Low Pathogenic Avian Influenza Virus Including Extract of Smilax china L. - Google Patents

Abstract

The present invention relates to a disinfectant composition for low pathogenic avian influenza virus, composed by containing: 95.5 parts by weight of purified water; 5.0 parts by weight of a Smilax china extract; 3.0 parts by weight of a Perilla frutescens leaf extract; 2.0 parts by weight of ionized mineral nanoparticles; 1.0 part by weight of sodium cyanide (NaOCN); 0.5 part by weight of glutaraldehyde (C_5H_8O_2); 0.5 part by weight of guar hydroxypropyltrimonium chloride; and 0.5 part by weight of citric acid (C_6H_8O_7). The disinfectant composition for low pathogenic avian influenza virus containing the Smilax china extract, the Perilla frutescens leaf extract, ionized mineral nanoparticles and the like according to the present invention not only has a very good disinfecting effect on low pathogenic avian influenza virus, but also has a little effect on environmental pollution.

Description

Process for Compositions of Disinfectants for Low Pathogenic Avian Influenza Virus Including Extract of Smilax china L.

The present invention relates to a method for preparing a low pathogenic avian influenza virus disinfectant composition containing a Tobokyeong extract and a low pathogenic avian influenza virus disinfectant composition prepared therefrom. More specifically, the method for producing a low pathogenic avian influenza virus disinfectant composition, and the low pathogenic avian influenza, characterized in that it comprises a Tobokyeong extract, perilla leaf extract and mineral particles ionized by electrolysis and nanoparticles as an active ingredient. The invention relates to a virus disinfectant composition.

Avian Influenza virus is a virus that infects poultry (chicken, turkey, duck, etc.), human influenza virus, swine influenza virus, horse influenza virus, and mainly enveloped virus. Depending on the degree of pathogenicity of the virus can be classified into low and high pathogenic avian influenza. Influenza virus types are classified into types A, B, and C according to the components of the virus (matrix, nucleoprotein). Cause disease.

The serotypes of influenza A are classified according to the two types of proteins (Hemagglutinin and Neuraminidase), which are H and N serotypes. The serotypes of influenza virus are expressed in their respective H and N serotypes, for example H9N2. Influenza viruses have 16 subtypes of H serotypes from H1 to H16, and 9 subtypes of N serotypes from N1 to N9. Influenza viruses can be present arithmetically when H and N are combined. There are 144 serotypes (16 × 9). Serotypes are represented by H3N2, H9N2 and the like, and have an important correlation with the pathogenicity of the virus.

Each serotype does not have a cross-immune response or is weak to prevent infection with other serotypes of influenza virus, ie, chickens immunized with H6N1 serotype virus cannot prevent infection with other serotypes of influenza virus such as H5N2. Vaccines are difficult to develop because influenza viruses of various serotypes have no antigen related to defense regardless of serotype. In addition, it is difficult to develop a timely disinfectant for bird flu, although domestic and global recurrent bird flu occurs every year. Therefore, there is an urgent need worldwide for the development of individual antiviral efficacy evaluations and compositions for each serotype that can be variously combined with H6N1, H5N1, H9N2, H5N6 and the like in advance. In addition, the evaluation of the efficacy of disinfectants on low-pathogenic and high-pathogenic viruses is also required.

As a related art in this regard, Patent Document 0001 below discloses "sterilization and virucidal composition improved by disinfecting by including citric acid and tartaric acid in trichophyllitis and malic acid," but this is a type of virus, avian influenza virus has been disclosed It is not specific. In addition, since the composition is made of natural materials, biodegradation by microorganisms is possible, so that it has little effect on environmental pollution, and has a disadvantage in that the disinfection effect against low pathogenic avian influenza virus is not excellent.

In addition, Patent Document 0002 discloses a "disinfection method for inactivating avian influenza virus by contacting a disinfectant with a drug containing a fumigant obtained from wood as an active ingredient." It is not. This is because the composition is made from natural materials, the load on the global environment is very low, there is no concern for the safety and hygiene of the human body and animals, and there is a disadvantage that the disinfection effect against the low pathogenic avian influenza virus is not excellent.

In addition, Patent Document 0003 discloses "a composition for disinfecting algae influenza, including sodium metasilicate and potassium carbonate," but this is only a very general disinfectant composition for disinfecting the surrounding birds and various livestock equipment and facilities, It is not a composition for disinfecting avian influenza virus that is not specific for a particular low pathogenic serotype.

Therefore, the present inventors, while studying the disinfectant for the low pathogenic avian influenza virus unlike the prior art, by mixing the ionized minerals of the Tobokyeong extract, Perilla persimmon leaf extract as an active ingredient with nano- and electrolysis by ultrasonic grinding as a active ingredient to a specific low-pathogenic serotype It was found that the disinfection effect is excellent to complete the present invention.

Korean Laid-Open Patent Publication No. 10-2009-0009442 (2009. 01. 13.) Japanese Laid-Open Patent Publication No. 2013-136610 (2012. 01. 17.) Korea Patent Publication No. 10-0772054 (2007. 10. 25.)

An object of the present invention is to provide a method for producing a low pathogenic avian influenza virus disinfectant composition containing the Tobokyeong extract.

The present invention comprises the steps of: 1) cutting the Tobok-ryeong to an appropriate size, extracting using ethanol as a solvent, filtration and freeze drying to obtain Tobok-Ryung extract; 2) a second step of cutting perilla leaves into an appropriate size, extracting using ethanol as a solvent, filtering and freeze drying to obtain perilla leaves extract; 3) a third step of calcining the shells, pulverizing and meshing, and then ultrasonically pulverizing and nanoning, followed by electrolysis to ionize the mineral particles; 4) purified water, Tobokyeong extract obtained in the first step, perilla leaf extract obtained in the second step, sodium cyanate (NaOCN), glutaraldehyde (C ₅ H ₈ O ₂ ), A fourth step of preparing a mixed solution by mixing guar hydroxypropyltrimonium chloride (Guar Hydroxypropyltrimonium Chloride) and citric acid (C ₆ H ₈ O ₇ ); 5) provides a method for producing a low pathogenic avian influenza virus disinfectant composition comprising; a fifth step of nano-pulverizing the mixed solution made in the fourth step so as not to precipitate.

In addition, the present invention 95.5 parts by weight of purified water Tobokyeong extract 5.0 parts by weight, perilla leaf extract 3.0 parts by weight, ionized mineral nanoparticles 2.0 parts by weight, sodium cyanate (NaOCN) 1.0 parts by weight, glutaraldehyde (C ₅ H ₈ O ₂ ) 0.5 parts by weight, guar hydroxypropyltrimonium chloride (Guar Hydroxypropyltrimonium Chloride), 0.5 parts by weight of citric acid (C ₆ H ₈ O ₇ ) It provides a low pathogenic avian influenza virus disinfectant composition.

Other specific examples for implementing the present invention are included in the following detailed description.

The method for producing a low pathogenic avian influenza virus disinfectant composition containing the Tobokyeong extract according to the present invention is a low pathogenic avian influenza virus disinfectant composition containing ionized various minerals obtained from Tobokyeong extract, perilla leaf extract and shellfish by electrolysis and nano-processing As a method for the preparation, there are various advantages such as excellent disinfection effect against low pathogenic avian influenza virus and little effect on environmental pollution.

Hereinafter, with reference to the method for producing a low pathogenic avian influenza virus disinfectant composition of the present invention will be described in detail preferred embodiments and physical properties measurement method. The present invention can be better understood by the following examples, the following examples are intended to be described in detail so that those skilled in the art can easily carry out the invention, This does not mean that the technical spirit and scope of the present invention is limited.

In the present invention, "disinfection" means killing pathogens in order to prevent infection or transmission of the disease, and in the present invention, the "microorganism" is included as an active ingredient in the low pathogenic avian influenza virus disinfectant composition of the present invention. It is a concept that includes all bacteria, fungi, yeasts and algae in which the constituent material exhibits disinfection effect.

Hereinafter, a method for preparing the low pathogenic avian influenza virus disinfectant composition of the present invention will be described in more detail.

Method for producing a low pathogenic avian influenza virus disinfectant composition according to an embodiment of the present invention is a method for producing a low pathogenic avian influenza virus disinfectant composition according to an embodiment of the present invention 1) extract the Tobokyeong using ethanol as a solvent, A first step of filtering and freeze-drying to obtain Tobokyeong extract; 2) a second step of cutting perilla leaves to an appropriate size, extracting using ethanol as a solvent, filtering and freeze drying to obtain perilla leaves extract; 3) a third step of sintering the shells, pulverizing and meshing and then ultrasonically pulverizing and nanoning, followed by electrolysis to ionize the mineral nanoparticles; 4) purified water, Tobokyeong extract obtained in the first step, perilla leaf extract obtained in the second step, sodium cyanate (NaOCN), glutaraldehyde (C ₅ H ₈ O ₂ ), A fourth step of preparing a mixed solution by mixing guar hydroxypropyltrimonium chloride (Guar Hydroxypropyltrimonium Chloride) and citric acid (C ₆ H ₈ O ₇ ); 5) a fifth step of nanofiberization so as not to precipitate by ultrasonic grinding the mixed solution made in the third step; characterized in that it comprises a.

The first step is a preparation step in the method for producing a low pathogenic avian influenza virus disinfectant composition of the present invention, by cutting the Tobokyeong to an appropriate size, using ethanol as a solvent and extract, filtered and freeze-dried Tobokyeongyeong extract It is a step to get.

The second step is to cut perilla leaves to an appropriate size, extract using ethanol as a solvent, filtered and freeze-dried to obtain perilla leaves extract.

Tobokyeong used in the present invention is a root stem of Smilax China L. which is a deciduous shrub of the liliaceae genus Liliaceae distributed in Korea, China, Japan, etc., and has been widely used in traditional herbal medicine or folk medicine. . Detoxification and dehumidification, arthralgia, muscle paralysis, diarrhea, dysentery, edema, disease, hepatitis, nausea, hemorrhoids, gastric cancer, esophageal cancer, etc. are known to have therapeutic effects.

The term "perilla perilla" refers to a leaf of a year-old herb belonging to the Lamiaceae, and the perilla leaves may be any perilla leaves, preferably domestic perilla leaves, which are commercially available in Korea.

Tobokyeong extract or perilla leaf extract of the present invention can be extracted at room temperature or warmed under the condition that the active ingredient is not destroyed, or minimized. The Tobokyeong extract or perilla leaf extract may be obtained by reflux extraction at 80 ℃, 50 minutes and 30 minutes, respectively, with 70Vol% ethanol, respectively.

The Tobokyeong extract or perilla leaf extract is preferably extracted by adding an extraction solvent of 2 to 20 times, preferably 5 to 15 times the dry weight of Tobokyeong or Perilla persimmon leaf, most preferably 10 times the weight. Although water is generally used as the extraction solvent, other organic solvents may be used in addition to water, if necessary. Organic solvents that may be used include, but are not limited to, C ₁ to C ₄ lower alcohols. Preferably ethanol and its aqueous solution are mentioned, Most preferably, 70 vol% ethanol. In the present invention, the "Tobokyeong extract" or "perilla perilla leaf extract" is preferably the Tobokyeong extract or perilla leaf extract prepared using the extracting solvent, the amount added in the disinfectant composition of the present invention is 5.0 parts by weight, 3.0 It is preferable to set it as weight part. Tobokyeong extract or perilla leaf extract extracted using an ethanol solvent may be filtered to remove foreign substances, such as to improve the quality to prevent deterioration, and freeze-dried for preservation.

The third step is the step of sintering the shells, pulverizing and meshing, and then ultrasonically pulverizing and nanonized, followed by electrolysis to ionize the mineral particles.

First, the shellfish is not limited because any kind of shellfish that can be obtained from the beach can be used, but it is preferable to use a scallop shell or cockle shells. The calcined mineral obtained by calcining the shellfish at 1,200 ° C. for 2 hours may be pulverized with a pulverizer, and may contain Mg, Na, K, P, S, Fe, and Zn as constituents thereof. It is preferable that the particle size is a powder of 30 ~ 300 mesh, and if the particle size is less than 30 mesh, the price is expensive and there is an economic problem, and if the particle size exceeds 300 mesh, a problem occurs that does not melt.

In addition, the ultrasonic grinding process of applying ultrasonic wave 25W three times for 10 minutes to the meshed mineral particles is first performed. 100 nm-sized nanoparticles were obtained through the ultrasonic grinding process, and the size of the particles did not change even when kept at room temperature for 7 days (see Table 1). After the nano-ization process through the ultrasonic pulverization process, while stirring the mineral particles at 100rpm in the state dissolved in water to increase the temperature slightly to 45 ~ 50 ℃ condition, and then to the (+) and (-) Electrolyte leads to electrolysis of 0.1 ~ 5V electricity for 3 ~ 5 times in a few minutes. Electrolysis of water in the state of dissolving mineral particles promotes ionization of minerals, thereby inhibiting precipitation of each mineral particle and maintaining an ionized state. At this time, the following reaction occurs in the cathode (Cathode) and the anode (Anode), the ionization of the minerals in this process proceeds.

(1) cathode _{(Cathode): 2H 2 O +} 2e - -> 2OH - + H 2

(2) The positive electrode ^{(Anode): 2OH - ->} H 2 O + 2e - + 1 / 2O 2

On the other hand, the nano- and electrolysis carried out in the above raises the solubility of water and minerals, causing the effect of maintaining solubility even after a considerable time (about 4 weeks or more) time after dissolution. As a result of experimenting with the aqueous solution of mineralized particles after the nano-ization and electrolysis separately, the nanoparticle size change was observed within the error range of less than 10% by maintaining 105 to 110 nm even after 270 days at room temperature. It can be seen that the relatively stable state (see Table 1). In addition, the addition amount in the disinfectant composition of the present invention is preferably 2.0 parts by weight.

Particle Size by Preservation Period of Mineral Particles Storage temperature Retention period (days) Particle size (nm) 25 ± 2 ℃ 0 100 7 101-103 Room temperature 30 100 60 101 90 102 180 104 240 108 270 110

The fourth step is purified water, the Tobokyeong extract obtained in the first and second steps, perilla leaf extract, sodium cyanate (NaOCN), glutaraldehyde (C ₅ H ₈ O ₂ ) to the ionized mineral nanoparticles of the third step , Guar Hydroxypropyltrimonium Chloride (Guar Hydroxypropyltrimonium Chloride), citric acid (C ₆ H ₈ O ₇ ) is a step of making a mixed solution. At this time, the mixing ratio of the constituents was 95.5 parts by weight of purified water 5.0 parts by weight of Tobokyeong extract, 3.0 parts by weight of perilla leaf extract, 2.0 parts by weight of ionized mineral nanoparticles, 1.0 parts by weight of sodium cyanate (NaOCN), glutaraldehyde (C ₅ H ₈ O ₂ ) 0.5 parts by weight of Guar Hydroxypropyltrimonium Chloride (Guar Hydroxypropyltrimonium Chloride) and 0.5 parts by weight of citric acid (C ₆ H ₈ O ₇ ) may be contained to form a mixed solution.

The sodium cyanate (NaOCN) is a white crystalline solid, which is used as an oxidizing agent in the disinfectant composition of the present invention to destroy proteins of viruses by oxidation. In addition, the addition amount in the disinfectant composition of the present invention is preferably 1.0 parts by weight.

Since glutaraldehyde (C ₅ H ₈ O ₂ ) is toxic, it should not be in direct contact with humans and livestock, and the amount is preferably 0.5 parts by weight. In addition, this is a good disinfecting effect even if some organic matters, but the expensive side has a disadvantage that takes a lot of disinfection costs to use in large quantities. This also means that the disinfection effect is maintained even without addition.

The citric acid (citric acid) is contained in the seed or juice of many plants as a free acid, and is widely used in various applications, mainly fruit flavor supplements, preservatives, pH regulators. It is safe for people and clothes, so the effect is good, but the penetration is weak, so the effect is very low in the presence of organic matter. The addition amount is preferably 0.5 parts by weight, which means that the disinfection effect is maintained even without addition.

In the fifth step, the mixed solution made in the fourth step is ultrasonically pulverized so as not to precipitate. It is preferable to help the solution of the mixture by nano-sonication by performing ultrasonic sonication process by repeatedly applying 5W ultrasonic wave three times for 10 minutes to the mixed solution prepared in the fourth step, and preventing the precipitate from occurring.

The first to fifth steps were carried out sequentially to 5.05.5 parts by weight of Tobokyeong extract, 9 parts by weight of perilla leaf extract, 2.0 parts by weight of ionized mineral nanoparticles, 1.0 parts by weight of sodium cyanate (NaOCN), glue Low pathogenic algae that does not precipitate by containing 0.5 parts by weight of taraldehyde (C ₅ H ₈ O ₂ ), 0.5 parts by weight of guar hydroxypropyltrimonium chloride, and 0.5 parts by weight of citric acid (C ₆ H ₈ O ₇ ) Flu virus disinfectant compositions can be prepared.

Meanwhile, the low pathogenic avian influenza virus disinfectant composition of the present invention may be prepared by further including additives such as a dispersant, a carrier, and other antimicrobial agents, in addition to the component, as long as the disinfecting effect of the component is not inhibited. have.

For example, in the case of dispersants, water, alcohols (e.g. methyl alcohol, ethyl alcohol, ethylene glycol, propylene glycol, diethylene glycol, glycerin, etc.), ketones (e.g. acetone, methyl ethyl ketone, etc.), ethers (e.g. Dioxane, tetrahydrofuran, cellosolve, diethylene glycol dimethyl ether, etc., aliphatic hydrocarbons (e.g. hexane, kerosene, etc.), aromatic hydrocarbons (e.g. benzene, toluene, xylene, naphthalene, methyl naphthalene, etc.), halogenated hydrocarbons (E.g., chloroform, carbon tetrachloride, etc.), acidamides (e.g., dimethyl formadiate, etc.), esters (e.g. methyl acetate ester, ethyl acetate ester, butyl acetate ester, fatty acid glycerin ester, etc.), nitrole (e.g. Acetonitrile, etc.), surfactants (higher alcohol sulfate esters, alkyl sulfonic acids, alkyl allyl sulfonic acids, quaternary ammonium salts, oxy alkylamines, fatty acids) Thermal switch, a polyalkylene oxide compound, anhydro-sorbitol compounds), but can be one or two or more selected from, but is not limited thereto.

Also in the case of carriers, clays (e.g. kaolin, bentonite, acid clays, etc.), talc (e.g. talc powders, feldspar powders, etc.), silicas (e.g. diatomaceous earth, silicic anhydrides, mica powders, etc.), alumina, sulfur powders and activated carbon One or two or more of them may be selected, but is not limited thereto.

The low pathogenic avian influenza virus disinfectant composition of the present invention may be prepared in liquid or solid phase, and the disinfectant composition of the present invention may be administered in any manner such as oral or parenteral. Oral dosage forms include tablets, pills, powders, solutions, and food forms. Parenteral dosage forms include injections, topical dosages (creams, ointments, etc.), suppositories, and sprays (if applied to plants). Etc. can be mentioned.

The disinfectant composition of the present invention is preferably administered in a topical manner, and is applied to the low-pathogenic avian influenza virus H9N2, and can be administered / sprayed in and around birds or colonies and breeding areas.

In addition, a tissue made of a mixed material of pulp and nonwoven fabric having a size of 20 cm × 10 cm in width and width is immersed in the disinfectant composition according to the present invention, and the disinfectant composition is allowed to soak into the tissue for 1 to 2 hours. An antiviral cleaning tissue impregnated with the disinfectant composition according to the invention can be prepared. In the case of cleaning the face and hands using the cleaning tissue prepared in this way, the antiviral effect was significantly improved compared to the case of washing the exposed skin using the conventional hand cleaner. Iii) it was confirmed by skin irritation test that no skin damage shape appeared.

Hereinafter, the present invention can be better understood by the following preparation examples and examples, the following preparation examples and examples are for illustrative purposes of the present invention and are not intended to limit the protection scope of the present invention.

<Production example>

  1. Experimental Materials

    In order to verify the antiviral effects of the test substance against avian influenza virus (AI virus), the experimental animals used 2-3 week old SPF (specific pathogen free) chickens and SPF fetal eggs, and the virus used was low pathogenic algal influenza (LPAI). -H9N2) was used.

  2. Tobokyeong extract manufacture

     Tobokyeong was purchased on the drug. After cutting 1000 g of Tobok-ryeong to an appropriate size, using 10L of 70Vol% ethanol as a solvent and extracted for 50 minutes at 80 ℃ using a reflux extractor. The extract was filtered through Whatman filter paper and 30.1 g of dry weight extract was obtained using a lyophilizer.

  3. Perilla Leaf Extract Manufacturing

     Perilla leaves were purchased at hypermarkets. 1000g of perilla leaves were cut to a suitable size, and then extracted with a reflux extractor at 80 ° C. for 30 minutes using 10L of 70Vol% ethanol as a solvent. The extract was filtered through Whatman filter paper and 32.2 g of dry weight extract was obtained using a lyophilizer.

  4. Preparation of Ionized Mineral Nanoparticles

     1) The scallops or cockle shells were calcined at 1,200 ° C. for 2 hours, and the calcined minerals were pulverized with a grinder to obtain a powder having a particle size of 30 to 300 mesh.

     2) The calcined minerals contained Mg, Na, K, P, S, Fe, Zn as constituents. Ultrasound of 25 W was repeatedly applied to the mesh-sized mineral particles three times for 10 minutes, and the nanoparticles obtained by grinding were electrolyzed to obtain ionized mineral nanoparticles.

<Example>

     In order to prepare a low pathogenic avian influenza virus disinfectant composition according to the present invention, the composition is uniformly mixed in the composition ratio shown in the following [Table 2] to make a mixed solution, and then repeatedly subjected to 25W ultrasonic wave three times for 10 minutes and pulverized the mixed solution. It was electrolyzed again to obtain an ionized nanomixture to obtain a disinfectant composition.

Ingredients and Ingredient Ratios Ingredient Component ratio (part by weight) Purified water 95.5 Tobokyeong Extract 5.0 Perilla Leaf Extract 3.0 Ionized mineral nanoparticles 2.0 Sodium cyanate 1.0 Glutaraldehyde 0.5 Guar hydroxypropyltrimonium chloride 0.5 Citric acid 0.5

Experimental Example Evaluation of Experimental Methods and Inhibitors

  1. Experiment Method

    In order to test the disinfecting effect of the disinfectant composition of the <Example> was tested according to the National Veterinary Research and Quarantine Service Notification No. 2016-91 [Disinfectant Effectiveness Test Guideline] and the results are shown in the following [Table].

  2. Evaluation of Inhibitors

   1) Each virus stock was diluted 1: 10 times, 1.5 ml ep tube was used, and 1 µg / ml TPCK in DMEM was used as the dilution medium.

   2) The test substance and the virus reaction were kept at 4 ℃ by storing all the test solution and reagents, ep tube was used.

     2-1) 450 μl of test substance and 100 μl of virus were mixed.

     2-2) The material control group was mixed with 450 µl of PBS and 100 µl of virus.

     2-3) After mixing, the mixture was reacted at 4 ° C. for 30 minutes, and mixed well every 10 minutes.

   3) 450 μl 1M Tris-HCl (pH 7.2) was added, and 1M Tris-HCl (pH 7.2) for neutralizing the test substance pH was used as a diluent.

   4) Viral titer evaluation was performed on the mixed solution, and a deep well plate was used.

      4-1) 10-fold step dilution was performed at a ratio of 100 µl of the mixed solution and 900 µl of the medium.

4-2) The dilution factor was 10 ⁻¹ , 10 ⁻² , 10 ⁻³ , 10 ⁻⁴ , 10 ⁻⁵ , 10 ⁻⁶ , and 100 μl of the cells was inoculated, and 8 replicates were performed for each dilution factor.

       Dilution medium: 1 μg / ml TPCK in DMEM

     * In the case of fertilized eggs 0.2ml was inoculated into the allantoic cavity, incubated for 5 days in a 37 ℃ incubator and tested every day, and the presence of virus growth was examined.

   5) The reaction was carried out for 1 hour at 37 ℃.

   6) After the reaction, the inoculum was removed.

   7) 200 ml of culture medium was added.

     Medium: 1 µg / ml TPCK in DMEM

8) Cultured in a 37 ℃ CO ₂ incubator, Carbapenemase production Enterobacteriaceae (CPE) was observed, the results are shown in the table below.

<Experimental Results> Evaluation of antiviral inhibitory ability against avian influenza virus

In the above table, in the case of pathogen control, fertilized egg infectivity was confirmed even at a dilution factor of 10 ⁸ or more. In addition, as criteria, for the disinfectant criteria 10 ^4. Suppressing ⁰ EID ₅₀ or more viral replication it was determined that the viral inhibitory effects. In other words, it was confirmed that the example composition of the present invention inhibited virus propagation by virus 10 ^4.0 EID ₅₀ or higher.

In the evaluation of inhibitory ability using cells, it was confirmed that the example composition of the present invention inhibits the virus, but the influenza virus used in the test had a proliferation in the embryonated eggs so that it was difficult to determine the exact inhibitory ability. Inhibition of proliferation was confirmed.

  <Comprehensive Analysis of Experiment Results>

In summary, the low pathogenic avian influenza virus disinfectant composition of the present invention is excellent in inhibiting ability against avian influenza virus using embryonated eggs and cells, as well as protecting birds from diseases caused by the virus, and side effects and residuals for drugs. It was evaluated that it can be used safely without concern about the safety of livestock products by drugs.

As mentioned above, although this invention was demonstrated by the limited Example, this invention is not limited by this and it should be described below by the person of ordinary skill in the art, and the following. Of course, various modifications and variations are possible within the scope of the claims.

Claims (15)

1) the first step of cutting the Tobokyeong to the appropriate size, extract using ethanol as a solvent, filtered and freeze-dried to obtain the Tobokyeong extract;
2) a second step of cutting perilla leaves to an appropriate size, extracting using ethanol as a solvent, filtering and freeze drying to obtain perilla leaves extract;
3) a third step of calcining the shells, pulverizing and meshing, and then ultrasonically pulverizing and nanoning, followed by electrolysis to ionize the mineral particles;
4) purified water, Tobokyeong extract obtained in the first step, perilla leaf extract obtained in the second step, sodium cyanate (NaOCN), glutaraldehyde (C ₅ H ₈ O ₂ ), A fourth step of preparing a mixed solution by mixing guar hydroxypropyltrimonium chloride (Guar Hydroxypropyltrimonium Chloride) and citric acid (C ₆ H ₈ O ₇ );
5) a fifth step of nano-sizing to prevent precipitation of the mixed solution made in the fourth step so as not to precipitate; the method of producing a low pathogenic avian influenza virus disinfectant composition comprising a.
The method according to claim 1,
In the first step, ethanol was added to 70 times the weight ratio of 10 times to Tobokyeong, 50 minutes at 80 ℃, ethanol was added 70 times the weight ratio to perilla leaf 10 times in the second step, at 80 ℃ 30 minutes each A reflux extraction method for producing a low pathogenic avian influenza virus disinfectant composition.
The method according to claim 2,
In the third step, the shellfish is a method of producing a low pathogenic avian influenza virus disinfectant composition is scallops or cockle shells.
The method according to claim 3,
In the third step, the firing is a method for producing a low pathogenic avian influenza virus disinfectant composition to mesh for 2 hours at 1,200 ℃, 30 to 300 mesh grinding.
The method according to claim 4,
In the third step, the nano-induced low pathogenic avian influenza virus disinfectant composition of the particle size is 100nm by repeating the ultrasonic wave of 25W three times for 10 minutes.
The method according to claim 5,
In the fourth step, the mixed solution is 95.5 parts by weight of purified water, 5.0 parts by weight of Tobok-ryong extract, 3.0 parts by weight of perilla leaf extract, 2.0 parts by weight of ionized mineral nanoparticles, 1.0 parts by weight of sodium cyanate (NaOCN), glutaraldehyde (C ₅ H ₈ O ₂ ) Preparation of a low pathogenic avian influenza virus disinfectant composition is 0.5 parts by weight, guar hydroxypropyltrimonium chloride (Guar Hydroxypropyltrimonium Chloride) 0.5 parts by weight, citric acid (C ₆ H ₈ O ₇ ) 0.5 parts by weight of the mixture Way.
The method according to claim 6,
In the fifth step, the mixed solution is a method for producing a low pathogenic avian influenza virus disinfectant composition to repeat the ultrasonication of 25W for 10 minutes, three times.
5.0 parts by weight of Tobokyeong extract, 3.0 parts by weight of perilla leaf extract, scallops or cockle shells were calcined at 1,200 ° C for 2 hours to 30 to 300 mesh, and then ultrasonically crushed and electrolyzed to 100 nm in 95.5 parts by weight of purified water. 2.0 parts by weight of the ionized mineral nanoparticles, 1.0 part by weight of sodium cyanate (NaOCN), glutaraldehyde (C ₅ H ₈ O ₂ ), 0.5 part by weight, 0.5 part by weight of guar hydroxypropyltrimonium chloride (Guar Hydroxypropyltrimonium Chloride) , Low pathogenic avian influenza virus disinfectant composition containing 0.5 parts by weight of citric acid (C ₆ H ₈ O ₇ ).
The method according to claim 8,
The Tobokyeong extract is a low pathogenic avian influenza virus disinfectant composition obtained by reflux extraction of Tobokyeong with 70Vol% ethanol at 80 ℃ for 50 minutes.
The method according to claim 9,
The perilla leaf extract is a low pathogenic avian influenza virus disinfectant composition obtained by reflux extraction of perilla leaves with 70Vol% ethanol at 80 ℃ for 30 minutes.
delete The method according to claim 10,
The ionizing mineral nanoparticles are low pathogenic avian influenza virus disinfectant composition containing all of Mg, Na, K, P, S, Fe, Zn as a component.
The method according to claim 12,
The low pathogenic avian influenza virus is H9N2 low pathogenic avian influenza virus disinfectant composition.
The method according to any one of claims 8 to 10, 12, 13,
The low pathogenic avian influenza virus disinfectant composition further comprises one or more additives selected from a dispersant, a carrier.
The method according to any one of claims 8 to 10, 12, 13,
Anti-viral cleaning tissue, characterized in that prepared by impregnating the low pathogenic avian influenza virus disinfectant composition.