KR20200009520A - Process for Compositions of Disinfectants for High Pathogenic Avian Influenza Virus Including nano- and ionic-Minerals - Google Patents

Abstract

The present invention relates to a high pathogenic (H5N6) avian influenza virus disinfectant composition composed by containing, in 95.5 parts by weight of purified water, 2.0 parts by weight of mineral particles, 1.0 parts by weight of sodium cyanate (NaOCN), 0.5 parts by weight of glutaraldehyde (C_5H_8O_2), 0.5 parts by weight of guar hydroxypropyl-trimonium chloride, and 0.5 parts by weight of citric acid (C_6H_8O_7). The high pathogenic (H5N6) avian influenza virus disinfectant composition containing minerals according to the present invention has an excellent disinfection effect on highly pathogenic (H5N6) avian influenza virus and has an advantage of hardly affecting environmental pollution.

Description

Process for Compositions of Disinfectants for High Pathogenic Avian Influenza Virus Including nano- and ionic-Minerals

The present invention relates to a method for preparing a high pathogenic avian influenza virus disinfectant composition containing ionized and nanonized minerals, and to a high pathogenic avian influenza virus disinfectant composition prepared therefrom. More specifically, the present invention relates to a method for producing a highly pathogenic avian influenza virus disinfectant composition comprising a variety of minerals ionized by nanolysis and electrolysis by ultrasonic pulverization, and a high pathogenic avian influenza virus disinfectant composition prepared therefrom. .

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 humans and animals, and 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 viruses that is not specific for a particular high pathogenic serotype.

In addition, Patent Document 0004 discloses "an antibody containing a wide range of neutralizing activity against influenza virus type A group 1," which is an antibody having neutralizing activity against subtype H1 and subtype H5 of influenza A virus of group 1 It's just about.

Accordingly, the present inventors, while studying a disinfectant for high pathogenic avian influenza virus unlike the prior art, it is found that the disinfection effect is excellent for a specific high-pathogenic serotype by mixing the mineralization ionized by nanolysis and electrolysis by ultrasonic grinding as an active ingredient. The present invention has been completed.

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.) Korean Unexamined Patent Publication No. 10-2010-0115346 (2010.10.27.)

The present invention aims to provide a highly pathogenic avian influenza virus disinfectant composition containing ionized and nanonized minerals.

The present invention comprises the steps of: 1) calcining shells, pulverizing and meshing, followed by ultrasonic pulverization and nanoning, followed by electrolysis to ionize mineral particles; 2) Purified water, sodium cyanate (NaOCN), glutaraldehyde (C ₅ H ₈ O ₂ ), guar hydroxypropyltrimonium chloride (citrate) in the ionized mineral nanoparticles of the third step, citric acid (C ₆ H ₈ O ₇ ) mixing the second step to form a mixed solution; 3) provides a method for producing a highly pathogenic avian influenza virus disinfectant composition comprising a; a third step of nano-pulverizing the mixed solution made in the second step so as not to precipitate.

In addition, the present invention is 95.5 parts by weight of purified water 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 ( It provides a highly pathogenic avian influenza virus disinfectant composition comprising 0.5 parts by weight of Guar Hydroxypropyltrimonium Chloride) and 0.5 parts by weight of citric acid (C ₆ H ₈ O ₇ ).

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

A method for producing a highly pathogenic avian influenza virus disinfectant composition containing ionized and nanonized minerals according to the present invention is a method for producing a highly pathogenic avian influenza virus disinfectant composition containing ionized various minerals obtained from shellfish by electrolysis and nano process. In addition, it has an excellent disinfection effect against high pathogenic avian influenza virus and has little effect on environmental pollution.

Hereinafter, preferred embodiments and physical property measurement methods will be described in detail regarding a method for preparing a highly pathogenic avian influenza virus disinfectant composition containing ionized and nanonized minerals of the present invention. The present invention can be better understood by the following examples, the following examples are intended to explain in detail enough to be able to easily carry out the invention of those skilled in the art to which the present invention pertains. 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 highly 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 a disinfecting effect.

Hereinafter, a method for preparing a highly pathogenic avian influenza virus disinfectant composition containing the ionized and nanonized minerals of the present invention will be described in more detail.

Method for producing a highly pathogenic avian influenza virus disinfectant composition containing ionized and nano-mineralized minerals according to an embodiment of the present invention is 1) firing shells, pulverized and meshed and then ultrasonically pulverized and nanoscaled, then electrolyzed minerals A first step of ionizing the nanoparticles; 2) Purified water, sodium cyanate (NaOCN), glutaraldehyde (C ₅ H ₈ O ₂ ), guar hydroxypropyltrimonium chloride (C ₆ ) to the ionized mineral nanoparticles of the first step, citric acid (C ₆ H ₈ O ₇ ) mixing the second step to form a mixed solution; 3) a third step of nano-sonication so as not to precipitate by ultrasonic grinding the mixed solution made in the second step; characterized in that it comprises a.

The first step is a preparation step in the method for producing a highly pathogenic avian influenza virus disinfectant composition containing the ionized and nano-mineralized mineral of the present invention, sintered shellfish, pulverized and meshed, and then ultrasonically pulverized and nanoscaled, Decomposing and ionizing 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 second step is purified water, the ionized mineral nanoparticles of the third step sodium cyanate (NaOCN), glutaraldehyde (C ₅ H ₈ O ₂ ), guar hydroxypropyltrimonium chloride (Guar Hydroxypropyltrimonium Chloride), citric acid (C ₆ H ₈ O ₇ ) is a step of mixing to form a mixed solution. At this time, 95.5 parts by weight of purified water, 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 ) 0.5 parts by weight of the mixture can be prepared.

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 third step, the mixed solution made in the second step is ultrasonically pulverized so as not to be precipitated. It is preferable to help the solution of the mixture by nano-sonication by applying a 5W ultrasonic wave three times for 10 minutes again to the mixture solution made in the second step to prevent the precipitate from occurring.

The first step to the fifth step was carried out sequentially to 2.0 parts by weight of ionized mineral nanoparticles, 9 parts by weight of sodium cyanate (NaOCN), 0.5 parts by weight of glutaraldehyde (C ₅ H ₈ O ₂ ) to 95.5 parts by weight of purified water It is possible to prepare a highly pathogenic avian influenza virus disinfectant composition comprising 0.5 parts by weight of guar hydroxypropyltrimonium chloride (Guar Hydroxypropyltrimonium Chloride), 0.5 parts by weight of citric acid (C ₆ H ₈ O ₇ ) to prevent precipitation.

On the other hand, the highly pathogenic avian influenza virus disinfectant composition containing the ionized and nano-mineralized mineral of the present invention, in addition to the above components, additives such as dispersants, carriers and other antimicrobial agents, in addition to the components, as long as they do not inhibit the disinfecting effect of the active ingredient It can be prepared by further comprising.

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.), acid amides (e.g. dimethyl formamide, etc.), esters (e.g. methyl acetate ester, ethyl acetate ester, butyl acetate ester, fatty acid glycerine 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 highly pathogenic avian influenza virus disinfectant composition containing the ionized and nanonized minerals 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 H5N6, which is a highly pathogenic avian influenza virus, and may 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 x 10 cm in width and length is put into the disinfectant composition according to the present invention and soaked for 1 to 2 hours so that the disinfectant composition sufficiently penetrates the tissue. 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 effect 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. H5N6) was used.

  2. 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 highly pathogenic (H5N6) bird flu virus disinfectant composition containing a mineral 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 repeated 25W ultrasonic waves three times for 10 minutes. The mixed solution obtained by pulverization was electrolyzed again to obtain an ionized nano mixed solution to obtain a disinfectant composition.

Ingredients and Ingredient Ratios Ingredient Component ratio (part by weight) Purified water 95.5 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 table below.

  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 inspected 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

  1. Evaluation of Inhibitory Activity against High Pathogenic Avian Influenza Virus Using Cells

In case of H5N6 avian influenza, the proliferation in cells was good, and the inhibitory effect on the substance could be clearly confirmed. In both of the cells, it was confirmed that the composition of the present invention inhibited the growth of the virus. .

<Comprehensive Analysis of Experiment Results>

In summary, the highly pathogenic avian influenza virus disinfectant composition containing perilla leaf extract of the present invention is not only excellent in inhibiting the H5N6 avian influenza virus using Caco-2 cells and MDCK cells, but also from the disease caused by the virus. It is evaluated that it can be used safely without concern about side effects to the drug and safety of the livestock product by the residual drug.

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 (12)

1) a first step of calcining shells, pulverizing and meshing, followed by ultrasonic pulverization and nanoning, followed by electrolysis to ionize mineral particles;
2) Purified water, sodium cyanate (NaOCN), glutaraldehyde (C ₅ H ₈ O ₂ ), guar hydroxypropyltrimonium chloride (C ₆ ) to the ionized mineral nanoparticles of the third step, citric acid (C ₆ H ₈ O ₇ ) mixing the second step to form a mixed solution;
3) a third step of nanofiberization so as not to precipitate by ultrasonic grinding the mixed solution made in the second step; a method for producing a highly pathogenic (H5N6) bird flu virus disinfectant composition comprising a.
The method according to claim 1,
In the first step, the shellfish is a scallop shell or cockle shell is a high pathogenic (H5N6) avian influenza virus disinfectant manufacturing method.
The method according to claim 2,
In the first step, the firing is 2 hours at 1,200 ℃, pulverization is a method of producing a highly pathogenic (H5N6) bird flu virus disinfectant composition to mesh to 30 ~ 300 mesh.
The method according to claim 3,
The method of producing a highly pathogenic (H5N6) avian influenza virus disinfectant composition in the first step is to repeat the nano-wave 25W three times for 10 minutes, three times the size of the particles.
The method according to claim 4,
In the second step, the mixed solution is 95.5 parts by weight of purified water, 2.0 parts by weight of ionized mineral nanoparticles, 1.0 part by weight of sodium cyanate (NaOCN), 0.5 part by weight of glutaraldehyde (C ₅ H ₈ O ₂ ), guar hydroxypropyl tree A method for producing a highly pathogenic (H5N6) bird flu virus disinfectant composition which is mixed at a ratio of 0.5 parts by weight of monium chloride (Guar Hydroxypropyltrimonium Chloride) and 0.5 parts by weight of citric acid (C ₆ H ₈ O ₇ ).
The method according to claim 5,
Method of producing a highly pathogenic (H5N6) avian influenza virus disinfectant composition in which the mixed solution in the third step is to nano-sized 25W ultrasonic wave for 10 minutes, three times.
95.5 parts by weight of purified water, 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) A high pathogenic (H5N6) bird flu virus disinfectant composition comprising 0.5 parts by weight and 0.5 parts by weight of citric acid (C ₆ H ₈ O ₇ ).
The method according to claim 7,
The mineral particles are obtained by calcining shellfish for 2 hours at 1,200 ℃ high pathogenic (H5N6) bird flu virus disinfectant composition.
The method according to claim 8,
The shellfish is a scallop clam or cockle shell is a high pathogenic (H5N6) bird flu virus disinfectant composition.
The method according to claim 9,
The ionized mineral nanoparticles of the high pathogenic (H5N6) bird flu virus disinfectant composition containing Mg, Na, K, P, S, Fe, Zn as a component.
The method according to any one of claims 7 to 10,
The high pathogenic (H5N6) bird flu virus disinfectant composition further comprises one or more additives selected from a dispersion, a carrier.
The method according to any one of claims 7 to 10,
Antiviral cleaning tissue, characterized in that prepared by impregnating the high pathogenic (H5N6) bird flu virus disinfectant composition.