KR20090009442A - Antimicrobial and antiviral composition having improved sterilizing power - Google Patents

Abstract

A composition comprising malic acid, triple salt, citric acid and tartaric acid is provided to improve antiseptic effects against virus and bacteria compared to the disinfectant 'cone -S' without an effect on the environmental contamination. The antibacterial and antiviral composition comprises 30-50 parts by weight of triple salt, 1-20 parts by weight of sodium hexameta phosphate, 1-10 parts by weight of malic acid, 1-10 parts by weight of citric acid and 1-10 parts by weight of tartari acid, and further comprises 1-35 parts by weight of dodecylsulfate sodium salt.

Description

Antimicrobial and antiviral composition having improved sterilizing power

The present invention relates to a bactericidal and virucidal composition with improved disinfection ability, and more particularly to a bactericidal and virucidal composition having improved disinfection power by including citric acid and tartaric acid in trichophytes and malic acid.

Recently, the pathogens in the livestock industry are avian influenza virus, foot-and-mouth disease virus, porcine cholera virus, salmonella, streptococci and the like.

In particular, avian influenza virus, which mainly causes poultry such as chickens and turkeys, is classified into three types of high pathogenicity, weak pathogenicity, and non-pathogenicity according to pathogenicity. High hospitality is classified as List A by the International Water Services Bureau (OIE), and as a type 1 animal infectious disease in Korea. The causative agent is type A virus, which has H serotype and N serotype. A total of 135 viral serotypes can exist by these two proteins. As of 2004, there have been 15 reported HAs and 9 NAs worldwide. Infection occurs mainly in direct contact with the secretions of birds, and is also transmitted by splashes, water, human feet, feed cars, appliances, equipment, and feces on the outside of eggs.

Symptoms vary depending on the pathogenicity of the infected virus, but usually include respiratory symptoms, diarrhea and a sharp decrease in egg production. Occasionally, cyanosis appears on the head of the crest, edema on the face, or feathers gather in one place.

The mortality rate varies from 0 to 100% depending on the pathogenicity. Newcastle disease, infectious laryngeal tracheitis, mycoplasma infection, etc. have similar symptoms.

Although it has not occurred worldwide since the 1930s, it began to occur in Europe such as Belgium and France in 1983, and since 2004, highly pathogenic avian influenza, including pharmacopathy, has occurred in various countries. High pathogenicity also affected humans, killing six people in Hong Kong in 1997 and 16 people in Vietnam in 2004. In Korea, avian influenza occurred in Eum, Chungcheongbuk-do, in December 2003, and spread throughout the country. However, it was confirmed that the disease was not transmitted to the human body because of its pathogenicity.

If bird flu occurs, most of the world's slaughterhouses will be slaughtered, and the producing countries will not be able to export poultry products.

Therefore, avian influenza virus can be said to be the leading role in the development of the livestock industry.

Foot-and-mouth disease occurs when livestock are infected with foot-and-mouth disease virus, which occurred in Korea and Japan in 2000, and has been occurring worldwide since 2001. The major regions are Africa, South America, Asia, and Eastern Europe. As of August 30, 2001, non-occurring countries recognized by OIE include North and Central American countries such as the United States, Canada, Panama, Norway (Western Europe), Australia, and New Zealand. (Ocean Province), Indonesia, Japan (Asia) and more than 50 countries, and the northwestern Colombia and Mindanao region of the Philippines are regionally recognized, and the countries that are vaccinated are Paraguay and Brazil and Colombia, which are recognized in some regions. In the case of Jeju Island, which was recognized only in Jeju Island, as of September 19, 2001, it was certified as a non-occurring country (cleaning country) that is vaccinating with France, the Netherlands and Ireland.

Due to the economic damage caused by the decline in animal productivity due to the foot-and-mouth disease, foot-and-mouth disease is considered to be a regulated disease in the international trade, and among the 15 kinds of diseases classified as Class A by the International Water Bureau (OIE), the foot-and-mouth disease is the first malignant epidemic. This is difficult to root out and there is no special treatment.

In order to develop the livestock industry, in addition to the above-mentioned pathogens, it is necessary to develop a wide range of safe disinfectants capable of freely disinfecting livestock and drinking water, as well as high sterilizing power and livestock disinfection for killing pathogens and viruses that cause various diseases.

Natural products mean natural products without adding artificial ones. Natural products classified as GRAS (Generally Recognized As Safe) are used as food additives because they are classified as natural additives in Korea without being regulated in terms of consumption or target food. . In foreign countries, it is a substance that can be freely used according to the user's intention without any special limitation, and it is used as a health food and medicine because of its excellent functionality.

Malic acid is a rich source of natural fruit and is generally recognized as safe (GRAS) by the US Food and Drug Administration (FDA). Malic acid is registered as a drug notification material by the KFDA and is used as a raw material for medicines. It has been used for a long time as an additive in soft drinks, canned fruits and vegetables, fruit juices, and candy / chewing gum.

Currently, we are making great efforts to develop disinfectants using natural materials all over the world, and commercially available disinfectants are Bayer Korea's 'Bercon-S' whose main ingredients are malic acid and three kinds of salts.

The 'Bercon-S' is the most sold as a foot-and-mouth disinfectant, and is the best-known disinfectant of the Purbright Institute, showing excellent killing effects against various pathogens such as viruses, bacteria, and fungi, as well as safety for young puppies, herbivore, newborn piglets, and newborn calves. This required livestock has been known as a safe disinfectant that can be sprayed directly onto the shaft. The disinfectant is 500 g of trisalt per kg. 15 g of sodium chloride. It contains 100 g of malic acid, which is diluted in 50 ~ 300 times of water for sterilization.

However, natural substances have to be developed and developed continuously, considering the increased resistance of pathogens over time due to various factors such as long-term use of chemicals and environmental pollution. Therefore, in the art, there is a need for the development of natural disinfectants capable of freely disinfecting shafts and drinking water, as well as high disinfecting power and barn disinfection, compared to currently available disinfectants capable of killing pathogens and viruses that cause various diseases.

Therefore, the present inventors have studied intensively to prepare a natural disinfectant having high sterilizing power and barn disinfection ability against various pathogens, and as a result, a composition containing citric acid and tartaric acid in malic acid and tricalcium at a specific ratio is higher than the current commercially available VERCON-S. The disinfecting power was remarkably improved and the stability was maintained and the present invention was completed.

An object of the present invention is to provide a bactericidal and virucidal composition containing citric acid and tartaric acid in a specific ratio in malic acid and trigeminal salt.

In order to achieve the above object, the present invention provides an antiseptic and bactericidal composition having improved disinfecting ability including trichloride, sodium hexametaphosphate, malic acid, citric acid and tartaric acid.

Hereinafter, the present invention will be described in detail.

In the bactericidal and virucidal composition according to the present invention, the content of the composition is 30 to 50 parts by weight of potasium monopersulfate, 1 to 20 parts by weight of hexametaphosphate, 1 to 10 parts by weight of malic acid, and 1 to 10 parts of citric acid. It is preferable to include a weight part and 1-10 weight part of tartaric acid.

Potassium monopersulfate, one of the essential components of the present invention, is a tribasic salt of potassium peroxymonosulfate, potassium sufate and potassium bisulfate, which is better known under the trade name of 'oxone ^® ' of DuPont acid, which is used as a main component of disinfectant. It is a substance, soluble in water, and non-chlorine oxidizer. Compared with chlorine-based oxidants, it does not produce a pungent odor and shows strong disinfection effect by strong oxidation.

The content of the trichloride is preferably 30 to 50 parts by weight. If the content of the trichloride is less than 30 parts by weight, there is a problem of weak sterilization and disinfection, and if it exceeds 50 parts by weight, it is too toxic to be harmful to humans or animals.

Sodium hexametaphosphate, one of the essential components of the present invention, is well soluble in water and can be hydrolyzed to regular phosphate in a high temperature aqueous solution, and the only aqueous solution of the polymerized phosphate exhibits the property of being neutral. It is generally used as a stabilizer for maintaining conservative properties of sausages, preventing fruit juice discoloration, pH buffers, cheese stabilizers, pectin coagulants, protein dispersants, and vegetables, fruits, and processed food-related products. It is known to suppress. In addition, when livestock farms, animal husbandry, and its surrounding facilities are metals, it serves to prevent or eliminate the corrosion of metals and to protect the surface of the metals to prevent corrosion of the metals.

In the present invention, the content of sodium hexametaphosphate is preferably 1 to 20 parts by weight. If the content of sodium hexametha phosphate is less than 1 part by weight, there is a problem in preventing and removing the corrosion of hardware, and if it exceeds 20 parts by weight, there is a problem of preventing the action of removing the corrosion of the metal.

Malic acid, one of the essential components of the present invention, is a substance widely known for its bactericidal and viricidal effects by acid, and generally recognized by the US Food and Drug Administration (FDA) as generally safe (GRAS). Also known as maleic acid or ungmul acid. The chemical formula of DL-Maleic acid is C ₄ H ₆ O ₅ , molecular weight 134.09, specific gravity 1.601, melting point 133 ° C, boiling point 150 ° C (decomposition). Malic acid is not deliquescent, and the acidity is about 20% stronger than citric acid, and it is rich in natural fruits such as apples and grapes. When acidic malic acid is added to a microorganism, the microorganism draws organic acids into the microorganism to maintain pH homeostasis. This mechanism acidifies the cytoplasm of the microorganisms, affects enzymatic reactions and mass transfer processes, and consumes energy for pH homeostasis, killing microbes. Malic acid also acts to relieve the odor and irritation of the disinfectant disinfectant.

In the present invention, the content of the malic acid is preferably 1 to 10 parts by weight. If the amount of the malic acid is less than 1 part by weight, there is a problem of mild relaxation, and if it exceeds 10 parts by weight, there is a problem in that the efficacy of the disinfectant and disinfectant is lowered.

Citric acid as an essential component of the present invention is an acid which is contained as a free acid in the seeds or juices of many plants, and is a component widely used for various purposes such as fruit flavor supplements, preservative synergists, and pH adjusters. In addition, tartaric acid is contained in tin precipitated when making wine, and is widely used as a soft drink in syrups and juices, and as a medicine for mixing purposes such as bases, buffers, and pH adjusters. None reported. They are listed on the Food Safety (GRSA) List, which is generally recognized as safe by the FDA. They are safe substances, act as a preservative and protect against decay by various organic substances. They are also accompanied by disinfectant and disinfectant components. Therefore, the drug is more strongly exerted when penetrated. Therefore, they serve to enhance the disinfecting power of the tribasic salt by acid by adding an appropriate amount to the tribasic salt.

In the present invention, the content of citric acid and tartaric acid is preferably 1 to 10 parts by weight independently. If the content of citric acid and tartaric acid is less than 1 to 10 parts by weight, there is almost no disinfection effect and anti-corruption effect, if the content exceeds 10 parts by weight there is a problem that the efficacy of the disinfectant and disinfectant is lowered.

In the bactericidal and virucidal composition according to the present invention, sodium dodecyl sulfonate may be further added to the composition to uniformly emulsify and disperse various components of the bactericidal and disinfectant, wherein the content of sodium dodecyl sulfonate is 1 to 35 It is preferable that it is a weight part.

In the bactericidal and virucidal composition according to the present invention, pigments, esters and the like may be further added to the composition to improve flavor and color.

The bactericidal and virucidal composition according to the present invention is a tribasic salt, sodium hexametaphosphate, sodium dodecylbenzenesulfonate, malic acid, sodium chloride and sulfa by adding citric acid and tartaric acid to potasium monopersulfate, sodium hexametaphosphate and malic acid. The bactericidal power of Bucon-S, a widely used disinfectant composed of sulphamic acid, was increased by about 2 to 4 times (see Tables 1 to 9). Therefore, the bactericidal and virucidal composition according to the present invention is made of a natural material and can be biodegraded by microorganisms, thus hardly affecting environmental pollution, and excellent disinfection effect against a wide range of bacteria and viruses even under high organic matter conditions. In addition, it can be usefully used for disinfecting shafts and drinking water.

The bactericidal and virucidal composition according to the present invention can sterilize Salmonella, Brucellella, Escherichia coli, Pasteurella, Staphylococcus, etc. as bacteria, and Newcastle Disease Virus, Avian Influenza Virus, Swine Ozeski Disease Virus, Pig Cholera virus, foot and mouth virus can be removed.

Hereinafter, the present invention will be described in more detail with reference to Examples. The above examples are for illustrating the present invention, but the scope of the present invention is not limited by these examples.

Example 1 Preparation of Bactericidal Composition 1

Triglyceride 500 g, malic acid 100 g, citric acid 15 g, tartaric acid 10 g, sodium hexametaphosphate 50 g and sodium dodecylbenzenesulfonate 319 g, with a small amount of red pigment and lemon flavor powder A disinfectant was prepared according to this procedure.

Example 2 Preparation of Bactericidal Composition 2

A disinfectant was prepared in the same manner as in Example 1 except that 500 g of trichloride, 100 g of malic acid, 100 g of citric acid, 50 g of tartaric acid, 125 g of hexamethaphosphate, and 25 g of sodium dodecylbenzenesulfonate.

< Experimental Example  1> sterilization effect

<1-1> Determination of the bactericidal effect of Salmonella

Salmonella cholearsuis (KCCM41598) as a representative strain of the general bacteria was distributed from the Korea Microorganism Conservation Center (KCCM) was used for the test.

The test strain was subcultured at 37 ° C. for 22-26 hours using autoclaved nutrient medium, and then the number of bacteria was measured by measuring turbidity according to the McFaland method, and more than 10 ⁸ CFU / mL were used.

The composition of Example 1 was divided into treatment 1, treatment 2, treatment 3, treatment 4, treatment 5, treatment 1, distilled water conditions (no organic matter), treatment 2, hard water conditions (low organic matter), treatment tools 3, organic / hard water conditions (high organic matter), treatment 4 is prepared as a control of treatments 2 and 3, treatment 5 is prepared as a control of treatment 1 and diluted with distilled water, hard water and 5% organic diluent for each treatment. It was. At this time, the dilution drainage is 1/100, 1/150, 1/200, 1/250, 1/300, 1/350, 1/400, 1/800, 1/900, 1/1000, 1/1100, 1 Diluted to / 1200 and 1/1400.

Hard water was dissolved in 0.15 g of CaCl ₂ and 0.139 g of MgCl ₂ 6H ₂ O in 1 L of distilled water and sterilized under high pressure and stored at 4 ° C. The dilute solution for bacteria was dissolved in hard water to contain 20% (w / v) yeast extract. It was then autoclaved and stored at 4 ° C. In dilution of the disinfectant and dilution of the bacteria, the dilution was prepared in distilled water, hard water, and 5% organic matter, and then pH was adjusted using 1 N NaOH and 1 N HCl to pH 7.0. The organic dilution solution for the virus was used by dissolving in sterile hard water to contain 5% fetal bovine serum.

Thereafter, 4 ml of the bacteria cultured at 37 ° C. were mixed in 96 ml of distilled water, hard water, and 5% organic matter dilution solution, and 2.5 ml of the mixed solution was taken out into a diluent solution of 4 ° C. diluted with distilled water, hard water and 5% organic matter dilution, respectively. Put and mix 1: 1. The reaction of the disinfectant and the cause of the disease was carried out at intervals of 1 minute in turn in each test tube, and reacted at 4 ° C. for exactly 30 minutes, and mixed every 10 minutes to allow sufficient reaction to occur.

After exactly 30 minutes of reaction, take out 1.0 ml immediately to neutralize the efficacy of the disinfectant and The mixture was added to 9.0 ml of neutralizing medium, mixed, and then mixed in five test tubes containing nutrient medium for each 0.1 ml of each disinfectant dilution, followed by incubation for 48 hours in a 37 ° C. incubator. Neutralizing medium for neutralizing the disinfectant component was used medium containing 5% inactivated horse serum in nutrient medium.

Experiments were performed on five nutrient media per diluent fold to determine the number of nutrient media in which bacteria were found.

Determination of bacterial growth was made as the dilution factor as the final disinfection dilution step in which two or more growths were not recognized in the nutrient medium of the five same disinfectant dilution factors.

In the above method, the bactericidal effect of the composition according to the present invention was measured and shown in Table 1.

Bactericidal Effect on Salmonella Disinfectant Dilution Drainage Treatment Treatment Zone 1 Treatment 2 Treatment Zone 3 Treatment Zone 4 Treatment Zone 5 1/100 0 0 0 5 5 1/150 0 0 0 5 5 1/200 0 0 3 5 5 1/250 0 0 5 5 5 1/300 0 0 5 5 5 1/350 0 0 5 5 5 1/400 0 0 5 5 5 1/800 0 0 5 5 5 1/900 0 One 5 5 5 1/1000 One One 5 5 5 1/1100 2 3 5 5 5 1/1200 4 4 5 5 5 1/1300 5 5 5 5 5 1/1400 5 5 5 5 5

As shown in Table 1, the composition according to the present invention has a bactericidal effect up to 1/1100 dilution when no organic matter or low concentration against Salmonella (treatment 1 or 2), and at high concentrations (treatment 3). It was confirmed that the bactericidal effect was up to 1/200 dilution.

<1-2> Determination of the bactericidal effect of staphylococci

Staphylococcus aureus to evaluate the disinfecting effect on certain bacteria (Staphylococcus aureus, KCCM40050) to accept pre-sale from South Korea microbial retention center was used for the test.

The test method was performed in the same manner as in <1-1>, and the results are shown in Table 2.

Bactericidal effect on staphylococci Disinfectant Dilution Drainage Treatment Treatment Zone 1 Treatment 2 Treatment Zone 3 Treatment Zone 4 Treatment Zone 5 1/100 0 0 0 5 5 1/150 0 0 0 5 5 1/200 0 0 One 5 5 1/250 0 0 2 5 5 1/300 0 0 5 5 5 1/350 0 0 5 5 5 1/400 0 0 5 5 5 1/800 0 0 5 5 5 1/900 0 0 5 5 5 1/1000 0 0 5 5 5 1/1100 One 2 5 5 5 1/1200 3 3 5 5 5 1/1300 5 5 5 5 5 1/1400 5 5 5 5 5

As shown in Table 2, the composition according to the present invention has a bactericidal effect up to 1/1200 dilution when no organic matter or low concentration against Staphylococcus aureus (Treatment 1 or 2), and at high concentrations (Treatment 3). It was confirmed that the bactericidal effect was up to 1/250 dilution.

<1-3> E. coli bactericidal effect measurement

Escherichia coli ( Esherichia coli) coli , KCCM11234) was distributed from Korea Microbial Preservation Center and used for the test.

The test method was carried out in the same manner as in <1-1>, and the results are shown in Table 3.

Bactericidal effect on E. coli Disinfectant Dilution Drainage Treatment Treatment Zone 1 Treatment 2 Treatment Zone 3 Treatment Zone 4 Treatment Zone 5 1/100 0 0 2 5 5 1/150 0 0 3 5 5 1/200 0 0 5 5 5 1/250 0 0 5 5 5 1/300 0 0 5 5 5 1/350 0 0 5 5 5 1/400 0 0 5 5 5 1/800 0 0 5 5 5 1/900 0 0 5 5 5 1/1000 2 3 5 5 5 1/1100 4 4 5 5 5 1/1200 5 5 5 5 5 1/1300 5 5 5 5 5 1/1400 5 5 5 5 5

As shown in Table 3, the composition according to the present invention has a bactericidal effect up to 1/1000 dilution factor when the organic matter is low or low concentration (Equipment 1 or 2) to E. coli, and at high concentrations (Treatment 3) It was confirmed that the sterilization effect up to / 150 dilution multiples.

<1-4> Measurement of bactericidal effect of Pasteurella

Pasteurella multocida type to evaluate the disinfecting effect against certain bacteria D ₄ ) was distributed from the National Veterinary Research and Quarantine Service and used for testing.

Plants were cultured in autoclaved brain and brain leachate (BHI broth) and subcultured at 37 ° C for 22-26 hours, followed by measurement of turbidity according to McFaland et al. A concentration of 10 ⁸ CUF / mL or more was used.

In order to neutralize the efficacy of the disinfectant after the reaction of the test method, 1.0 ml was immediately taken out and The mixture was placed in 9.0 ml of neutralizing medium, mixed, 0.1 ml of each disinfectant dilution was added to 5 test tubes containing BHI broth, and mixed for 48 hours in a 37 ° C. incubator. The neutralizing medium for neutralizing the disinfectant component was BHI. Except for using a medium containing 5% inactivated horse serum broth was carried out in the same manner as <1-1> and the results are shown in Table 4.

Bactericidal Effect on Pasteurella Disinfectant Dilution Drainage Treatment Treatment Zone 1 Treatment 2 Treatment Zone 3 Treatment Zone 4 Treatment Zone 5 1/100 0 0 0 5 5 1/150 0 0 One 5 5 1/200 0 0 3 5 5 1/250 0 0 3 5 5 1/300 0 0 5 5 5 1/350 0 0 5 5 5 1/400 0 0 5 5 5 1/800 0 0 5 5 5 1/900 0 0 5 5 5 1/1000 0 2 5 5 5 1/1100 2 3 5 5 5 1/1200 5 5 5 5 5 1/1300 5 5 5 5 5 1/1400 5 5 5 5 5

As shown in Table 4, the composition according to the present invention has a bactericidal effect up to 1/1100 dilution when no organic matter or low concentration of E. coli (treatment 1 or 2), and at a high concentration (treatment 3) 1 It was confirmed that the sterilization effect up to / 250 dilution multiples.

<1-5> Determination of the bactericidal effect of Brucella

To evaluate the disinfection efficacy of certain bacteria, Brucella ovis , ATCC25840) were used by the National Veterinary Research and Quarantine Service, and the strains were inoculated in 5% bovine serum contained brucella broth containing autoclaved 5% bovine serum and then incubated at 37 ° C. and 5% CO ₂ incubator. and then for 72 h subculture, by measuring the turbidity of the bacterial counts were determined according to methods such as McFaland, it was used in the tests that the concentration of bacteria 10 ⁸ CFU / mL or more.

When the test is finished, take out 1.0ml immediately to neutralize the efficacy of the disinfectant, 37 ℃ The mixture was placed in 9.0 ml of neutralizing medium, mixed, and then mixed in 5 test tubes containing Brucella medium for 0.1 ml of each disinfectant dilution, followed by incubation for 72 hours in a 37 ° C. 5% CO ₂ incubator. Neutralizing medium was carried out in the same manner as in <1-1> except that the culture medium containing 5% inactivated horse serum in Brussela medium and the results are shown in Table 5.

Brucella  For bactericidal effect Disinfectant Dilution Drainage Treatment Treatment Zone 1 Treatment 2 Treatment Zone 3 Treatment Zone 4 Treatment Zone 5 1/100 0 0 0 5 5 1/150 0 0 0 5 5 1/200 0 0 2 5 5 1/250 0 0 4 5 5 1/300 0 0 5 5 5 1/350 0 0 5 5 5 1/400 0 0 5 5 5 1/800 0 0 5 5 5 1/900 0 0 5 5 5 1/1000 One 2 5 5 5 1/1100 4 3 5 5 5 1/1200 5 5 5 5 5 1/1300 5 5 5 5 5 1/1400 5 5 5 5 5

As shown in Table 5, the composition according to the present invention has a bactericidal effect up to 1/1000 dilution factor when the organic matter is low or low concentration (Equipment 1 or 2) to E. coli, and at high concentrations (Treatment 3) It was confirmed that the sterilization effect up to / 200 dilution multiples.

< Experimental Example  2> Killer virus  effect

<2-1> Avian Influenza Virus of Killer virus  Effect measurement

Avian influenza virus H9N2 (MS96 strain), which was distributed by the National Veterinary Research and Quarantine Service, was used to evaluate the disinfection efficacy of the virus against the disinfectant. Virus strains were inoculated into the intimal cavity of 10-day-old SPF embryonated eggs and subcultured. At the time when the vitality of the propagated virus was the maximum (viral titer 10 ^7.0 / mL or more) it was read and used for the test.

The composition of Example 1 was divided into treatment 1, treatment 2, and treatment 3 to distilled water conditions (no organic matter) in treatment 1, hard water conditions (low organic matter) to treatment 2, organic matter / hard water conditions to treatment 3 Distilled water, hard water, and 5% organic diluents were prepared according to the treatment process, and the dilution factor was 1/200, 1/400, 1/600, 1/800, 1/1000, 1 Diluted to / 1200, 1/1400, 1/1600, 1/1800 and 1/2000.

Subsequently, 1.0 mL of the urinary coagulant solution containing virus was mixed with distilled water, hard water and organic diluent (5% fetal bovine serum) according to 24 mL of treatment, and 2.5 mL was taken at 1 minute intervals, distilled water, hard water, and 5%. After mixing the test tube containing a diluent of 4 ℃ diluted with organic diluent, and reacted for exactly 30 minutes at 4 ℃, mixed every 10 minutes to allow a sufficient reaction. After the reaction of the disinfectant was terminated, to neutralize the efficacy of the disinfectant immediately neutralized by mixing one portion of the disinfectant solution with one portion of 50% immobilized fetal bovine serum. Subsequently, the neutralized solution was diluted with stock solution, 10 ⁻¹ , 10 ⁻² , 10 ⁻³ , 10 ^−4, and 10 ⁻⁵ using PBS, and 0.2 mL was added into the intimal cavity of five 10-ylan SPF-grown eggs per dilution factor Inoculation was incubated for 5 days in an incubator at 37 ° C. Eggs were taken daily, and embryonated eggs that died within 24 hours of inoculation were considered as accidental deaths and excluded from test results. Virus presence and virus titer were finally determined by performing hemagglutination.

The virus content was calculated using the Karber method of the following equation (1).

Where L _{1 is} the Log value of the lowest dilution measured,

L is the difference in log values between dilutions

S is the sum of the natural death percentage of virus in each dilution.)

The experiment was repeated three times, and the measurement results are shown in Table 6.

Against avian influenza virus Killer virus  effect Treatment Disinfectant Dilution Drainage Neutralization dilution factor (positive water) EID ₅₀ Log reduction Stock solution 10 ^-1 10 ^-2 10 ^-3 10 ^-4 10 ^-5     Treatment tool 1 1/200 0 0 0 0 0 0 ≤10 ^0.5 ≥4.1 1/400 0 0 0 0 0 0 ≤10 ^0.5 ≥4.1 1/600 0 0 0 0 0 0 ≤10 ^0.5 ≥4.1 1/800 0 0 0 0 0 0 ≤10 ^0.5 ≥4.1 1/1000 3 0 0 0 0 0 10 ^0.1 4.0 1/1200 4 3 One 0 0 0 10 ^1.1 3.0 1/1400 5 5 3 One 0 0 10 ^2.3 1.8 1/1600 5 5 4 2 0 0 10 ^2.7 1.4 1/1800 5 5 5 5 3 0 10 ^4.1 0.0 1/2000 5 5 5 5 3 0 10 ^4.1 0.0     Treatment 2 1/200 0 0 0 0 0 0 ≤10 ^0.5 ≥4.3 1/400 0 0 0 0 0 0 ≤10 ^0.5 ≥4.3 1/600 0 0 0 0 0 0 ≤10 ^0.5 ≥4.3 1/800 0 0 0 0 0 0 ≤10 ^0.5 ≥4.3 1/1000 2 One 0 0 0 0 10 ^0.1 4.2 1/1200 3 3 One 0 0 0 10 ^0.9 3.4 1/1400 5 5 3 0 0 0 10 ^2.1 2.2 1/1600 5 5 5 2 0 0 10 ^2.9 1.4 1/1800 5 5 5 5 3 One 10 ^4.3 0.0 1/2000 5 5 5 5 3 One 10 ^4.3 0.0     Processing Unit 3 1/200 0 0 0 0 0 0 ≤10 ^0.5 ≥4.3 1/400 0 0 0 0 0 0 ≤10 ^0.5 ≥4.3 1/600 3 2 2 0 0 0 10 ^0.9 3.4 1/800 4 4 2 0 0 0 10 ^1.5 2.8 1/1000 5 4 4 One 0 0 10 ^2.3 2.0 1/1200 5 5 3 2 0 0 10 ^2.5 1.8 1/1400 5 5 4 2 2 0 10 ^3.1 1.2 1/1600 5 5 5 5 3 One 10 ^4.3 0.0 1/1800 5 5 5 5 3 One 10 ^4.3 0.0 1/2000 5 5 5 5 3 One 10 ^4.3 0.0 Pathogen control 5 5 5 4 3 2 10 ^4.3 Distilled Water Control 5 5 5 4 3 One 10 ^4.1

As shown in Table 6, the composition according to the present invention has a virucidal effect up to 1/1000 dilution when no organic matter or low concentration (a treatment 1 or a treatment 2) against avian influenza virus, and a high concentration (treatment 3) ) Has a virucidal effect up to 1/400 dilution.

<2-2> of Newcastle Disease Virus Killer virus  Effect measurement

To evaluate the antiseptic efficacy of the virus, Newcastle Disease Virus (Lasota strain) was distributed from the National Veterinary Research and Quarantine Service and used for the test.

After the reaction, except for dilution to 10 ^-1 , 10 ^-2 , 10 ^-3 , 10 ^-4 , 10 ^-5 and 10 ^-6 and carried out in the same manner as in <2-1> and the results are shown in Table 7 Indicated.

Against Newcastle disease virus Killer virus  effect Treatment Disinfectant Dilution Drainage Neutralization dilution factor (positive water) EID ₅₀ Log reduction 10 ^-1 10 ^-2 10 ^-3 10 ^-4 10 ^-5 10 ^-6     Treatment tool 1 1/200 0 0 0 0 0 0 ≤10 ^0.5 ≥5.3 1/400 0 0 0 0 0 0 ≤10 ^0.5 ≥5.3 1/600 0 0 0 0 0 0 ≤10 ^0.5 ≥5.3 1/800 0 0 0 0 0 0 ≤10 ^0.5 ≥5.3 1/1000 2 One 0 0 0 0 10 ^1.1 4.2 1/1200 3 3 0 0 0 0 10 ^1.7 3.6 1/1400 4 3 One 0 0 0 10 ^2.1 3.2 1/1600 5 5 2 0 0 0 10 ^2.9 2.4 1/1800 5 5 5 5 3 One 10 ^5.3 0.0 1/2000 5 5 5 5 2 2 10 ^5.3 0.0     Treatment 2 1/200 0 0 0 0 0 0 ≤10 ^0.5 ≥5.3 1/400 0 0 0 0 0 0 ≤10 ^0.5 ≥5.3 1/600 0 0 0 0 0 0 ≤10 ^0.5 ≥5.3 1/800 0 0 0 0 0 0 ≤10 ^0.5 ≥5.3 1/1000 0 0 0 0 0 0 ≤10 ^0.5 ≥5.3 1/1200 3 One One 0 0 0 10 ^1.5 3.8 1/1400 4 2 One 0 0 0 10 ^1.9 3.4 1/1600 4 4 3 One 0 0 10 ^2.9 2.4 1/1800 5 5 5 5 2 0 10 ^4.9 0.4 1/2000 5 5 5 5 3 One 10 ^5.3 0.0     Processing Unit 3 1/200 0 0 0 0 0 0 ≤10 ^0.5 ≥5.3 1/400 2 0 0 0 0 0 10 ^0.9 4.4 1/600 2 2 One 0 0 0 10 ^1.5 3.8 1/800 2 2 One 0 0 0 10 ^1.5 3.8 1/1000 5 3 One One 0 0 10 ^2.5 2.8 1/1200 5 5 3 2 0 0 10 ^3.5 1.8 1/1400 5 5 3 2 One 0 10 ^3.7 1.6 1/1600 5 5 3 4 One 0 10 ^4.1 1.2 1/1800 5 5 5 5 3 One 10 ^5.3 0.0 1/2000 5 5 5 5 3 One 10 ^5.3 0.0 Pathogen control 5 5 5 5 3 One 10 ^5.3 Distilled Water Control 5 5 5 5 3 One 10 ^5.3

As shown in Table 7, the composition according to the present invention has a virucidal effect up to 1/1000 dilution when no organic matter or low concentration (New treatment disease 1 or 2 treatment) against Newcastle disease virus, high concentration (process 3) It was confirmed that there is a virucidal effect up to 1/400 dilution factor.

<2-3> of swine Ozeski disease virus Killer virus  Effect measurement

To evaluate the antiseptic efficacy of the virus, the pig Ojesky disease virus (Yangsan strain) was distributed by the National Veterinary Research and Quarantine Service and used for the test.

Passive cultures were used in PK-15 cell lines to activate the swine Ozesky's disease virus. The content of the published virus was tested in advance and used for the test of 10 ^7.0 / mL or more. Diluted to 10 ⁻¹ , 10 ⁻² , 10 ⁻³ , 10 ⁻⁴ , 10 ^−5, and 10 ⁻⁶ , and 5 wells per dilution in 96 well tissue culture plates in which PK-15 monolayers were formed. 100 μl of the neutralized reaction solution was inoculated, incubated for 5 days at 37 ° C. after inoculation, and the result of <2-1> The results were shown in Table 8 by the same method.

About swine Ozeski disease virus Killer virus  effect Treatment Disinfectant Dilution Drainage Neutralization dilution factor (positive water) EID ₅₀ Log reduction 10 ^-1 10 ^-2 10 ^-3 10 ^-4 10 ^-5 10 ^-6     Treatment tool 1 1/200 0 0 0 0 0 0 ≤10 ^0.5 ≥5.3 1/400 0 0 0 0 0 0 ≤10 ^0.5 ≥5.3 1/600 0 One 0 0 0 0 10 ^0.7 4.6 1/800 2 One 0 0 0 0 10 ^1.1 4.2 1/1000 2 2 0 0 0 0 10 ^1.3 4.0 1/1200 5 5 3 0 0 0 10 ^3.1 2.2 1/1400 5 5 One 2 0 0 10 ^3.1 2.2 1/1600 5 5 3 2 One 0 10 ^3.7 1.6 1/1800 5 5 5 4 3 2 10 ^5.3 0.0 1/2000 5 5 5 4 3 2 10 ^5.3 0.0     Treatment 2 1/200 0 0 0 0 0 0 ≤10 ^0.5 ≥5.7 1/400 0 0 0 0 0 0 ≤10 ^0.5 ≥5.7 1/600 2 One 0 0 0 0 10 ^1.1 4.6 1/800 3 2 One 0 0 0 10 ^1.7 4.0 1/1000 5 3 One 0 0 0 10 ^2.3 3.4 1/1200 5 5 2 0 0 0 10 ^2.9 2.8 1/1400 5 5 2 0 0 0 10 ^2.9 2.8 1/1600 5 5 5 3 0 0 10 ^4.1 1.6 1/1800 5 5 5 4 3 2 10 ^5.3 0.4 1/2000 5 5 5 4 4 3 10 ^5.7 0.0     Processing Unit 3 1/200 0 0 0 0 0 0 ≤10 ^0.5 ≥5.7 1/400 2 One 0 0 0 0 10 ^1.1 4.6 1/600 5 2 0 0 0 0 10 ^1.9 3.8 1/800 4 3 One 0 0 0 10 ^2.1 3.6 1/1000 5 5 One 0 0 0 10 ^2.7 3.0 1/1200 5 5 4 One 0 0 10 ^3.5 2.2 1/1400 5 5 5 2 0 0 10 ^3.9 1.8 1/1600 5 5 4 3 One 0 10 ^4.1 1.6 1/1800 5 5 5 4 4 2 10 ^5.5 0.2 1/2000 5 5 5 4 4 3 10 ^5.7 0.0 Pathogen control 5 5 5 4 4 3 10 ^5.7 Distilled Water Control 5 5 5 4 3 2 10 ^5.3

As shown in Table 8, the composition according to the present invention has a virucidal effect up to a 1/1000 dilution factor when the organic matter is low or low concentration (treatment 1 or treatment 2) against the swine Ozeski disease virus, and when the concentration is high (treatment treatment) 3) was confirmed to have a virucidal effect up to 1/400 dilution.

<2-4> of porcine cholera virus Killer virus  Effect measurement

Porcine cholera virus (LOM strain) was distributed from the National Veterinary Research and Quarantine Service and used for testing to evaluate the antiseptic efficacy of the virus.

Passive cholera virus was used by subcultured in PK-15 cell line, and the content of the published virus was tested in advance and 10 ^7.0 / mL or more was used for the test, and 1.0ml of the virus solution was used in 19 mL of <2-1>. After dilution with distilled water, hard water and organic diluent (5% fetal bovine serum), take 2.5 mL at 1 minute intervals. The mixture was mixed in a test tube and then reacted at 4 ° C. for exactly 30 minutes, and mixed every 10 minutes to allow sufficient reaction to occur. The control group used distilled water instead of disinfectant solution. Subsequently, in order to neutralize the efficacy of the disinfectant after the reaction, the reaction solution was immediately mixed and mixed in a neutralizing solution (a cell culture solution containing 10% FBS) at 37 ° C in the same amount.

Neutralizing solution was diluted 10 ^-1 , 10 ^-2 , 10 ^-3 , 10 ^-4 , 10 ^-5 and 10 ^-6 using cell culture medium, and 50 μL of the diluted solution was added to 5 PK-15 monolayer cells per dilution. Was inoculated in a 96 well tissue culture plate formed, and incubated for 5 days in a CO ₂ incubator at 37 ℃. After removing the culture medium, the cells were fixed, and the presence of virus and virus titer were finally determined by evaluating the presence of porcine cholera antigen using CSFV E2 monoclonal antibody conjugated with FITC.

The test was repeated three times, and the results are shown in Table 9.

Against swine cholera virus Killer virus  effect Treatment Disinfectant Dilution Drainage Neutralization dilution factor (positive water) EID ₅₀ Log reduction 10 ^-1 10 ^-2 10 ^-3 10 ^-4 10 ^-5 10 ^-6     Treatment tool 1 1/200 0 0 0 0 0 0 ≤10 ^0.5 ≥5.1 1/400 0 0 0 0 0 0 ≤10 ^0.5 ≥5.1 1/600 0 0 0 0 0 0 ≤10 ^0.5 ≥5.1 1/800 2 0 0 0 0 0 10 ^0.9 4.2 1/1000 2 One 0 0 0 0 10 ^1.1 4.0 1/1200 5 3 2 0 0 0 10 ^2.5 2.6 1/1400 5 4 4 2 0 0 10 ^3.5 1.6 1/1600 5 5 5 4 2 2 10 ^5.1 0.0 1/1800 5 5 5 3 3 2 10 ^5.1 0.0 1/2000 5 5 5 4 4 0 10 ^5.1 0.0     Treatment 2 1/200 0 0 0 0 0 0 ≤10 ^0.5 ≥5.3 1/400 0 0 0 0 0 0 ≤10 ^0.5 ≥5.3 1/600 2 One 0 0 0 0 10 ^1.1 4.2 1/800 2 One 0 0 0 0 10 ^1.1 4.2 1/1000 2 2 0 0 0 0 10 ^1.3 4.0 1/1200 5 4 3 0 0 0 10 ^2.9 2.4 1/1400 5 4 3 2 0 0 10 ^3.3 2.0 1/1600 5 5 4 3 One 0 10 ^4.1 1.2 1/1800 5 5 5 4 2 2 10 ^5.1 0.2 1/2000 5 5 5 4 3 2 10 ^5.3 0.0     Processing Unit 3 1/200 0 0 0 0 0 0 ≤10 ^0.5 ≥5.3 1/400 2 One 0 0 0 0 10 ^1.1 4.2 1/600 2 3 2 0 0 0 10 ^1.9 3.4 1/800 4 3 3 0 0 0 10 ^2.5 2.8 1/1000 5 2 3 0 0 0 10 ^2.5 2.8 1/1200 5 4 3 0 0 0 10 ^2.9 2.4 1/1400 5 5 4 2 0 0 10 ^3.7 1.6 1/1600 5 5 5 3 2 0 10 ^4.5 0.8 1/1800 5 5 5 5 2 2 10 ^5.3 0.0 1/2000 5 5 5 4 3 2 10 ^5.3 0.0 Pathogen control 5 5 5 4 3 2 10 ^5.3 Distilled Water Control 5 5 5 4 3 One 10 ^5.1

As shown in Table 9, the composition according to the present invention has a virucidal effect up to 1/1000 dilution when no organic matter or low concentration (treatment 1 or treatment 2) with respect to porcine cholera virus, and high concentration (treatment 3) ) Was confirmed to have a virucidal effect up to 1/400 dilution.

Therefore, the bactericidal and virucidal composition according to the present invention can be used to disinfect by dilution of 400 times when a large amount of organic matter is present, and when the organic matter is less than 1000 times, it is widely used as a conventional disinfectant 'Bercon- Because it shows much better sterilization and virucidal effect than S 'disinfection (50 ~ 300 times dilution), it can be useful as various animal disinfectant, barn disinfectant, foot and mouth, bird flu prevention disinfectant.

As described above, since the bactericidal and virucidal compositions according to the present invention are made of natural materials and biodegradable by microorganisms, they have little effect on environmental pollution. It is much more effective in disinfecting a wide range of bacteria and viruses than S ', so it can be used not only for livestock sterilization but also for sterilization and negative sterilization.

Claims (6)

A disinfectant bactericidal and virucidal composition comprising trigeminal salt, sodium hexametaphosphate, malic acid, citric acid and tartaric acid. According to claim 1, wherein the content of the composition comprises 30 to 50 parts by weight of three kinds of salt, 1 to 20 parts by weight of hexametaphosphate, 1 to 10 parts by weight of malic acid, 1 to 10 parts by weight of citric acid, and 1 to 10 parts by weight of tartaric acid. Disinfection and improved bactericidal and killer virus composition, characterized in that. The antiseptic power and bactericidal composition of claim 1, further comprising sodium dodecylsulfonic acid salt. According to claim 3, The content of the sodium dodecyl sulfonate is 1 to 35 parts by weight, characterized in that the disinfection power and bactericidal composition improved. The bactericidal and virucidal composition of claim 1, wherein the bacterium is at least one selected from the group consisting of Salmonella, Brucella, Escherichia coli, Pasteurella, and Staphylococcus. According to claim 1, wherein the virus is at least one selected from the group consisting of avian influenza virus, Newcastle disease virus, swine Ozeski disease virus and swine cholera virus, foot-and-mouth virus, bactericidal and improved antimicrobial composition.