KR101857736B1 - Composition of chlorine disinfections free of chlorine smell - Google Patents

Abstract

More particularly, the present invention relates to a chlorine-based disinfecting and disinfecting composition, and more particularly, to a chlorine- And a component selected from the group consisting of sodium peroxide, sulfamic acid, and a mixture thereof. The chlorine-based disinfectant is excellent in efficacy of the chlorine-based disinfectant and effectively removes the chlorine odor and is environmentally friendly, safe for workers and animals, To a chlorine-based disinfecting and disinfecting composition which is effectively applied to a malicious infectious disease.

Description

Technical Field [0001] The present invention relates to a chlorine-based disinfectant composition for chlorine removal,

The present invention relates to a chlorine-based disinfecting and disinfecting composition comprising chlorine-based disinfecting disinfectant and chlorine-based disinfecting and disinfecting composition containing chlorine-based disinfecting disinfectant and a component selected from sodium peroxide, sulfamic acid, ≪ / RTI >

The chlorine-based disinfectant as a sterilizing disinfectant exhibits a powerful disinfecting power by inhibiting the permeability of the bacterial cell membranes, oxidizing the protoplasmic components, inhibiting the enzyme reaction, and in fact, the neutrophils (Neutrophils) (HOCl) is known to destroy bacteria by producing hypochlorous acid (HOCl).

Used for sterilization in the form of ^- chlorine-based sanitizers often Locks known as sodium hypochlorite (NaOCl) or chlorine dioxide (ClO _2), chlorine gas (Cl _2), hypochlorous acid (HOCl), hypochlorite ions (OCl) .

On the other hand, sodium dicarboxylate sodium (NaDCC) is a component showing about 100 times more effective than hypochlorous acid, and does not adversely affect the environment because it is 100% biodegradable and does not adversely affect the environment. As synergistic action with sodium carbonate as a foaming agent, It is known to promote the disinfection effect.

On the other hand, NaDCC disinfectants have superior productive power such as excellent disinfection effect, safe use and direct livestock application, but the demand of the anti-virus and livestock farmers was limited due to the generation of chlorine smell during use. Accordingly, these NaDCC disinfectants may be used as an ideal disinfectant for livestock only if the effect of deodorization and odor removal is added in addition to the wide effectiveness of sanitizing disinfection, and if the residual odor due to the disinfectant is removed.

Korean Patent No. 10-1054947

Accordingly, it is an object of the present invention to provide a chlorine-based disinfecting and disinfecting composition in which the chlorine-based disinfectant is effectively removed and the chlorine odor is effectively removed while maintaining the excellent effect of the chlorine-based disinfectant.

It is another object of the present invention to provide a chlorine-based disinfectant composition which is environmentally friendly, safe for workers and animals, and can be effectively applied to malignant infectious diseases such as foot-and-mouth disease and AI.

In order to accomplish the above object, the present invention provides a chlorine-based disinfectant; And a component selected from the group consisting of sodium peroxide, sulfamic acid, and mixtures thereof.

According to the present invention, there is provided a chlorine-based disinfectant which is effective in removing chlorine odor effectively while maintaining and improving the effectiveness of a chlorine disinfectant, and is environmentally friendly, safe for workers and animals, and can effectively apply malignant infectious diseases such as foot- can do.

Hereinafter, the present invention will be described in detail.

The present invention relates to a chlorine-based sanitizing disinfectant; And a component selected from the group consisting of sodium peroxide, sulfamic acid, and mixtures thereof.

Sodium peroxide, sulfamic acid, or a mixture thereof is used in a chlorine-based disinfecting disinfectant to stabilize a chlorine-based compound by substituting free chlorine in water, thereby reducing a chlorine-containing odor generated from the chlorine-based compound.

The sodium or sulfamic acid may be used alone in a chlorine-based disinfecting disinfectant, but it is most preferable to use sodium peroxide and a sulfamic acid together in order to more effectively remove chlorine and remove residual chlorine after use of the disinfectant. At this time, it is preferable to mix the sodium peroxide and the sulfamic acid in a weight ratio of 1: 10: 2 to 20 in order to exhibit optimal chlorine scavenging effect.

The content of sodium peroxide, sulfamic acid, or a mixture thereof may be appropriately controlled depending on the type of the chlorine-based compound used in the chlorine-based disinfectant, for example, 10 to 50 parts by weight per 100 parts by weight of the composition And preferably 20 to 40 parts by weight. If the content is less than 10 parts by weight, chlorine odor may occur. If the content is more than 50 parts by weight, the disinfecting power may be lowered.

The type of the chlorine-based disinfecting disinfectant is not limited as long as it is a compound used in a chlorine-based disinfecting disinfectant for the purpose of decolorizing, bleaching, sterilizing, disinfecting, etc. For example, sodium dichloroisocyanurate (NaDCC), trichloroisocyanurate Trichloroisocyanuric acid, calcium hypochlorite, chlorine water, sodium hypochlorite, sodium hypochlorite and the like can be used. Sodium dicarboxylate, hypochlorous acid or the like is preferably used for the development of the disinfecting effect, and sodium dicarboxylate is most preferably used.

The chloric sterilizing disinfectant may be contained in an amount of 50 to 80 parts by weight based on 100 parts by weight of the total composition.

It is needless to say that the chlorine-based disinfecting and disinfecting composition of the present invention can sterilize and disinfect bacterium, virus and the like by applying it to all the objects to which the conventional chlorine-based disinfecting and disinfecting agent is used. In particular, the chlorine-based disinfectant of the present invention is applied to an animal and is used as an antiseptic agent such as Clostridium perfringens such as Salmonella genus such as Salmonella typhimurium, Brucella genus such as Brucella ovis , Bacterium such as bacteria of the genus Tyridium, or variants thereof; Avian influenza virus (Avian influenza virus, AI), Newcastle Disease virus (Newcastle disease virus, ND), porcine epidemic diarrhea virus (Porcine epidemic diarrhea virus, PED) , porcine respiratory and reproductive syndrome virus (Porcine respiratory and reproductive syndrome virus, PRRS) , Foot and mouth disease virus (FMD), or the like, or a variant thereof.

In addition, the present invention may include an acid preparation, a basic preparation and the like, which are used in conventional chlorine-based disinfecting disinfectants, if necessary.

The kind of the acid preparation is not limited as long as it is a component used in a conventional disinfectant, and for example, adipic acid, citric anhydride and the like can be used.

The acid preparation may be included in an amount of 10 to 40 parts by weight, preferably 20 to 30 parts by weight, based on 100 parts by weight of the total composition.

The type of the basic agent is not limited as long as it is a component used in a conventional disinfectant, for example, sodium bicarbonate, anhydrous sodium carbonate, sodium hydrogen carbonate, sodium hydroxide, sodium percarbonate and the like can be used.

The basic agent may be included in an amount of 5 to 25 parts by weight, preferably 10 to 20 parts by weight, based on 100 parts by weight of the total composition.

Further, the present invention may further include a component that does not affect the action of sodium peroxide, sulfamic acid, etc., and is effective in removing chlorine. The ingredients that can be added include ascorbic acid, hydrogen peroxide, sodium bicarbonate, sodium carbonate, citric acid and the like. Can be used.

According to the present invention, chlorine-based disinfecting disinfectants including basic compounds, acid formulations and basic formulations are used together with sodium peroxide, sulfamic acid or a mixture thereof to effectively remove the chlorine odor, which has been a problem of the conventional chlorine- It is safe and easy to use it in an animal to which an operator or disinfectant is applied. Moreover, since the chlorine odor does not remain even after the disinfectant is used, the range of use can be extended widely.

That is, the chlorine scavenging disinfectant of the present invention has a wide range of disinfection effectiveness and chlorine scavenging efficacy, so that it can be exported to various countries where malignant infectious diseases such as foot-and-mouth disease and AI are well developed in Korea.

Hereinafter, the present invention will be described in more detail with reference to examples. These embodiments are for purposes of illustration only and are not intended to limit the scope of protection of the present invention.

Example 1. Production of chlorine-based disinfectant 1

A chlorine disinfectant was prepared by mixing 650 g of sodium dichloroisocyanurate, 250 g of sulfamic acid and 100 g of sodium peroxide.

Example 2. Production of chlorine-based disinfectant 2

Chlorinated disinfectant was prepared by mixing 600 g of sodium dichloroisocyanurate, 270 g of sulfamic acid, 100 g of sodium bicarbonate and 30 g of anhydrous sodium carbonate.

Comparative Example 1. Chlorine-based disinfectant

Aniguard Co., Ltd., consisting of 500 g of sodium dichloroisocyanurate, 240 g of adipic acid, 220 g of sodium bicarbonate and 40 g of anhydrous sodium carbonate, was used.

Experimental Example 1. Chlorine measurement of the disinfectant

The sanitizing and disinfecting agents of Examples 1 and 2 and Comparative Example 1 were diluted with water at a magnification of 1: 10-500 and chlorine was measured using MINIMAX-XP-Cl ₂ , a portable Cl ₂ measuring instrument of Honeywell, The results are shown in Table 1 below.

As shown in Table 1, chlorine-based compounds were used in the disinfectant disinfectants of Examples 1 and 2, in which sodium peroxide, sulfamic acid, or a mixture thereof was added according to the present invention, , And the chlorine-based disinfectant, Comparative Example 1, exhibited chlorine odor irrespective of the dilution ratio.

From these results, it was found that the chlorine-based disinfectant of the present invention containing sodium peroxide, sulfamic acid or a mixture thereof was safe to use without chlorine scavenging.

Experimental Example 2. Confirmation of chlorine removal mechanism

The chlorine scavenging mechanism of the disinfectant of Example 1 or 2 was confirmed, and the results are shown in Table 2 below.

division Reaction formula NaDCC C ₃ Cl ₂ N ₃ NaO ₃ + H ₂ O -> C ₃ H ₃ N ₃ O ₃ + HOCl Example 1 H ₂ NSO ₃ H + Na ₂ O ₂ + C ₃ Cl ₂ N ₃ NaO ₃ + H ₂ O
H ₂ NSO ₃ H + Na ₂ O ₂ + C ₃ H ₃ N ₃ O ₃ + HOCl
→ H ₂ NO ₂ + Na ₂ SO ₃ + H ₂ O + C ₃ Cl ₂ N ₃ NaO ₃ Example 2 H ₂ NSO ₃ H + C ₃ Cl ₂ N ₃ NaO ₃ + NaHCO ₃ + Na ₂ CO ₃ + H ₂ O
H ₂ NSO ₃ H + NaHCO ₃ + Na ₂ CO ₃ + C ₃ H ₃ N ₃ O ₃ + HOCl
→ Na ₂ CO ₃ + C ₃ Cl ₂ N ₃ NaO ₃ + H ₂ O + NaSO ₃

The sodium dicarboxylate sodium (NaDCC) used as the chlorine compound in the present invention forms HOCl which is a cause of chlorine scavenging as the diluted chloric acid when reacting with water. However, in the case of the chlorine-based disinfectant of the present invention, as shown in Table 2, HOCl, which is a causative substance of chlorine odor including sodium peroxide, sulfamic acid or a mixture thereof, was not formed. Respectively.

Experimental Example 3: Effective Dilution Scale of Bacterial Disinfection

3-1. Salmonella typhimurium Sterilization disinfection power for

For bacterial culture, high pressure sterilized Nutrient agar was added to Salmonella typhimurium ) and cultured at 37 ± 2 ° C for 20 hours. The cultured Salmonella typhimurium was inoculated into a test tube containing 20 mL of Nutrient broth and incubated with shaking at 37 ± 2 ° C. for 20 hours. The cultured bacteria were kept at 37 ° C until just before use, and the concentration of bacteria used was more than 1 × 10 ⁸ CFU per mL.

The sterilization disinfectants of Examples 1 to 2 and Comparative Example were immersed in a water-containing solution (1 L of distilled water, CaCl ₂ 0.305 g of MgCl ₂ .6H ₂ O and 0.139 g (w / v) of MgCl ₂ .6H ₂ O) and organic matter (prepared to contain yeast 5% (w / v) in hard water). The sterilization disinfectant of Example 1 was diluted at 4, 100, 4,500, 5,000, 5,600, and 6,200 magnifications using hard water and diluted at 70, 75, 8, 95, and 100 magnifications using organic matter. The disinfectant of Example 2 was diluted with water at 4, 100, 4,500, 5,000, 5,600, and 6,200 magnifications and diluted with organic matter to 60, 70, 75, 85, and 95 magnifications. In addition, the disinfectant of the comparative example was diluted at a magnification of 12,000, 13,500, 15,000, 16,500, and 18,500 using hard water and diluted with an organic material at a magnification of 60, 70, 75, 85 and 95.

Then, 4 mL of the bacteria cultured at 37 째 C were mixed with 96 mL of dilute hydrochloric acid or 5% organic material, respectively. 2.5 mL of the mixed solution was added to 2.5 mL of the disinfectant solution (4 DEG C) of each of the disinfectant solutions of Examples 1 and 2 and Comparative Example, mixed and reacted at 4 DEG C for exactly 30 minutes. At this time, each test tube treatment was performed at intervals of 1 minute, and shaking was performed every 10 minutes during the reaction. In the pathogen control group, hard water without sterilant diluent was used.

Immediately after the reaction was completed, 1 mL was taken out and mixed in 9 mL of neutralization medium (containing 5% (v / v) of horse serum unmixed in Nutrient broth) preliminarily placed at 37 ° C. Next, 5 mL of each disinfectant dilution step was prepared, and 0.1 mL of each was inoculated into a Nutrient broth tube and cultured in a 37 ° C incubator for 48 hours.

3-2. Brucella ovis Sterilization disinfection power for

Brucella ovis was plated on high pressure sterilized 5% Sheep blood agar and cultured at 37 ± 2 ° C for 20 hours. The cultured Brucella ovis was inoculated into a test tube containing 20 mL of Brain heart infusion broth and incubated at 37 ± 2 ° C. for 20 hours with shaking. The cultured bacteria were kept at 37 ° C until just before use, and the concentration of bacteria used was more than 1 × 10 ⁸ CFU per mL.

The sterilization disinfectants of Examples 1 to 2 and Comparative Example were immersed in a water-containing solution (1 L of distilled water, CaCl ₂ 0.305 g of MgCl ₂ .6H ₂ O and 0.139 g (w / v) of MgCl ₂ .6H ₂ O) and organic matter (prepared to contain yeast 5% (w / v) in hard water). The disinfectant of Example 1 was diluted with water at 2,500, 2,800, 3,000, 3,500, and 4,000, and diluted with organic materials at 80, 90, 10, 11, and 12 magnifications. The sterilization disinfectant of Example 2 was diluted with water at a dilution ratio of 1,600, 1,800, 2,000, 2,200, and 2,500, and diluted with organic matter at a ratio of 60, 70, 75, 85, and 95. In addition, the disinfectant of Comparative Example was diluted with water at 4, 100, 4,500, 5,000, 5,600, and 6,200, and diluted with organic matter at 40, 45, 50, 55 and 62 magnifications.

Then, 4 mL of the bacteria cultured at 37 째 C were mixed with 96 mL of dilute hydrochloric acid or 5% organic material, respectively. 2.5 mL of the mixed solution was added to 2.5 mL of each of the disinfectant diluents (4 ° C.) of Examples 1 and 2 and Comparative Example, and the mixture was reacted at 4 ° C. for exactly 30 minutes. At this time, each test tube treatment was performed at intervals of 1 minute, and shaking was performed every 10 minutes during the reaction. In the pathogen control group, hard water without sterilant diluent was used.

Immediately after the reaction was completed, 1 mL was taken out and mixed in 9 mL of neutralization medium (containing 5% (v / v) of horse serum unmixed in Nutrient broth) preliminarily placed at 37 ° C. Next, 5 mL of each disinfectant dilution step was prepared, 0.1 mL of each was inoculated into a Brain heart infusion broth tube, and then cultured in a 37 ° C. incubator for 120 hours.

3-3. Clostridium perfringens Sterilization disinfection power for

For culturing of bacteria, Clostridium perfringens was plated on high pressure sterilized 5% Sheep blood agar and incubated in an anaerobic culture apparatus at 37 ± 2 ° C for 20 hours. The cultured Clostridium pertussis was inoculated into a test tube containing 20 mL of NIH thioglycollate broth and cultured in an anaerobic culture incubator at 37 賊 2 属 C for 20 hours with shaking. The cultured bacteria were kept at 37 ° C until just before use, and the concentration of bacteria used was more than 1 × 10 ⁸ CFU per mL.

The sterilization disinfectants of Examples 1 to 2 and Comparative Example were immersed in a water-containing solution (1 L of distilled water, CaCl ₂ 0.305 g of MgCl ₂ .6H ₂ O and 0.139 g (w / v) of MgCl ₂ .6H ₂ O) and organic matter (prepared to contain yeast 5% (w / v) in hard water). The sterilization disinfectant of Example 1 was diluted with water at 2,000, 2,300, 2,500, 2,700, and 3,000 magnifications, and the organic matter was diluted at 25, 27, 30, 33, and 37 magnifications. The disinfectant of Example 2 was diluted with water at a dilution rate of 2,300, 2,500, 2,700, 3,000, and 3,300 and diluted with organic matter at 25, 27, 30, 33, and 37 magnifications. In addition, the disinfectant of the comparative example was diluted with water at a ratio of 6,000, 7,000, 7,500, 8,500, and 9,500, and diluted with organic matter at 25, 27, 30, 33, and 37 magnifications.

Then, 4 mL of the bacteria cultured at 37 째 C were mixed with 96 mL of diluted water or 5% organic material, respectively, according to the purpose. 2.5 mL of the mixed solution was added to 2.5 mL of each of the disinfectant diluents (4 ° C.) of Examples 1 and 2 and Comparative Example, mixed and reacted at 4 ° C. for exactly one hour. At this time, each test tube treatment was performed at intervals of 1 minute, and shaking was performed every 10 minutes during the reaction. In the pathogen control group, hard water without sterilant diluent was used.

Immediately after the reaction was completed, 1 mL was taken out and mixed in 9 mL of neutralization medium (prepared to contain 5% (v / v) of horse serum unmixed in NIH thioglycollate broth) previously placed at 37 ° C and mixed. Next, 5 mL of each disinfectant dilution step was prepared, and 0.1 mL of each was inoculated into NIH thioglycollate broth tube and incubated for 48 hours in an anaerobic incubator at 37 ° C.

3-4. Experiment result

After the cultivation of 3-1 to 3-3, the growth of each bacterium was visually observed, and the dilution magnification of the disinfectant which did not multiply four or more bacteria in the nutrient medium of five identical disinfectant dilution magnification was judged as the final effective dilution magnification And the results are shown in Table 3 below.

As shown in Table 3, the chlorine-based disinfectant of the present invention is Salmonella typhimurium), Orbis brucellosis (Brucella ovis and Clostridium perfringens . It was confirmed that, compared with Comparative Example 1, effective disinfecting power was obtained at a dilution ratio similar to that of Comparative Example 1, even without chlorination at all there was.

Experimental Example 4. Derivation of Effective Dilution Factor for Vaccine Disinfection

4-1. Avian influenza virus (AI) Disinfection power for H9N2

Frozen Avian influenza virus (AI) was dissolved and stored at 4 ℃ for a short time until immediately before use.

The sterilization disinfectants of Examples 1 to 2 and Comparative Examples were immersed in hard water (containing 0.305 g of CaCl ₂ and 0.139 g (w / v) of MgCl ₂ .6H ₂ O in 1 L of distilled water) and an organic material (fetal calf serum 5 % (v / v)) as follows. The sterilization disinfectant of Example 1 was diluted at 3,000, 4,000 and 5,000 magnifications using light water and diluted at 1,800, 2,000 and 2,200 magnifications using organic matter. The sterilization disinfectant of Example 2 was diluted with water at 6,000, 7,000, and 8,000 magnifications and diluted with organic matter at 1,600, 1,800, and 2,000 magnifications. The disinfectant of Comparative Example was diluted with water at 6,000, 7,000, and 8,000 magnifications and diluted with organic matter at 900, 1,100, and 1,400 magnifications.

Thereafter, 1.0 mL (1 × 10 1 to 1 × 10 ⁷ EID ₅₀ / mL) of virus solution stored at 4 ° C. was added to 19 mL of hard water or organic matter, which is a virus dilution solution, and mixed. 2.5 mL of the virus solution was added to each test tube containing the same amount of the disinfectant solution of Examples 1 and 2 and the comparative example, respectively (5 mL in total), and reacted at 4 ° C for exactly 30 minutes. At this time, each test tube treatment was performed at intervals of 1 minute, and shaking was performed every 10 minutes during the reaction. The pathogen control group used hard water without sterilant diluent, and the toxic control group used light water diluted disinfectant diluent.

Immediately after the reaction was completed, 1 mL was taken out and mixed in 5 mL neutralization medium (prepared so as to contain 10% (v / v) fetal bovine serum unmixed with PBS) previously placed at 37 ° C and mixed. Then, the neutralized solution was diluted with PBS to a concentration of 10 ^-1 , 10 ^-2 , 10 ^-3 , 10 ^-4 , and 10 ^-5 , and 5 dilutions per dilution were prepared. 10 days in the developmental field.

After inoculation, the cells were incubated at 37 ° C for 5 days, and the cells were sacrificed every day. The dead cells within 24 hours of inoculation were regarded as accidents and excluded from the test results. All the inoculated dead cells within 24 hours of inoculation within 5 days were stored at 4 ℃.

After 5 days of inoculation, all of the surviving bovine blastocysts and dead cells from 4 ° C were collected from the urethral lumen and the hemagglutination reaction was carried out using 1% chicken erythrocytes. The virus content was calculated using the Karber method.

The pathogen control group was tested in the light water condition without the disinfectant and the neutralization reaction step showed that the activity of the pathogen was 2 × 10EID ₅₀ / mL or more. In the toxic control group, it was confirmed that no bacterial toxicity by the disinfectant disinfectant occurred.

4-2. Newcastle disease virus (ND) Disinfection power against LsSota

Newcastle disease virus (ND), a cryopreserved Newcastle disease virus , was dissolved and stored at 4 ° C for a short time until immediately before use.

The sterilization disinfectants of Examples 1 to 2 and Comparative Examples were immersed in hard water (containing 0.305 g of CaCl ₂ and 0.139 g (w / v) of MgCl ₂ .6H ₂ O in 1 L of distilled water) and an organic material (fetal calf serum 5 % (v / v)) as follows. The disinfectant of Example 1 was diluted with water at 4,000, 6,000, and 8,000 magnifications and diluted with organic matter at 600, 1,000, and 1,400 magnifications. The disinfectant of Example 2 was diluted with water at 3,000, 4,000, and 6,000 magnifications and diluted with organic matter at 600, 1,000, and 1,400 magnifications. In addition, the disinfecting disinfectant of the comparative example was diluted with water at 6,000, 8,000, and 12,000 magnifications and diluted with organic matter at 500, 600 and 1,000 magnifications.

Thereafter, 1.0 mL (1 × 10 1 to 1 × 10 ⁷ EID ₅₀ / mL) of virus solution stored at 4 ° C. was added to 19 mL of hard water or organic matter, which is a virus dilution solution, and mixed. 2.5 mL of the virus solution was added to each test tube containing the same amount of the disinfectant solution of Examples 1 and 2 and the comparative example, respectively (5 mL in total), and reacted at 4 ° C for exactly 30 minutes. At this time, each test tube treatment was performed at intervals of 1 minute, and shaking was performed every 10 minutes during the reaction. The pathogen control group used hard water without sterilant diluent, and the toxic control group used light water diluted disinfectant diluent.

Immediately after the reaction was completed, 1 mL was taken out and mixed in 5 mL neutralization medium (prepared so as to contain 10% (v / v) fetal bovine serum unmixed with PBS) previously placed at 37 ° C and mixed. Then, the neutralized solution was diluted with PBS to a concentration of 10 ^-1 , 10 ^-2 , 10 ^-3 , 10 ^-4 , and 10 ^-5 , and 5 dilutions per dilution were prepared. 10 days in the developmental field.

After inoculation, the cells were incubated at 37 ° C for 5 days, and the cells were sacrificed every day. The dead cells within 24 hours of inoculation were regarded as accidents and excluded from the test results. All the inoculated dead cells within 24 hours of inoculation within 5 days were stored at 4 ℃.

After 5 days of inoculation, all of the surviving bovine blastocysts and dead cells from 4 ° C were collected from the urethral lumen and the hemagglutination reaction was carried out using 1% chicken erythrocytes. The virus content was calculated using the Karber method.

The pathogen control group was tested in the light water condition without the disinfectant and the neutralization reaction step showed that the activity of the pathogen was 2 × 10EID ₅₀ / mL or more. In the toxic control group, it was confirmed that no bacterial toxicity by the disinfectant disinfectant occurred.

4-3. Porcine epidemic diarrhea virus (PED)

Before testing, 200 μL of Vero cell (using α-MEM containing antibiotic and antimycotic as a culture medium) was dispensed in a 96-well plate at 2 × 10 ⁵ cells / mL and incubated at 37 ° C in 5% CO ₂ Lt; / RTI >

The sterilization disinfectants of Examples 1 to 2 and Comparative Examples were immersed in hard water (containing 0.305 g of CaCl ₂ and 0.139 g (w / v) of MgCl ₂ .6H ₂ O in 1 L of distilled water) and an organic material (fetal calf serum 5 % (v / v)) as follows. The disinfectant of Example 1 was diluted with water at a ratio of 30,000, 40,000, and 50,000, and diluted with organic matter at a ratio of 400, 800 and 1,200. The sterilization disinfectant of Example 2 was diluted with water at 30,000, 40,000, and 50,000 magnifications and diluted with organic matter at 800, 1,200, and 1,600 magnifications. In addition, the sterilizing disinfectant of the comparative example was diluted with water at 30,000, 40,000 and 50,000 magnifications, and diluted with organic matter at 100, 200 and 400 magnifications.

Thereafter, 1.0 mL (1 × 10 1 to 1 × 10 ⁷ EID ₅₀ / mL) of virus solution stored at 4 ° C. was added to 19 mL of hard water or organic matter, which is a virus dilution solution, and mixed. 2.5 mL of the virus solution was added to each test tube containing the same amount of the disinfectant solution of Examples 1 and 2 and the comparative example, respectively (5 mL in total), and reacted at 4 ° C for exactly 30 minutes. At this time, each test tube treatment was performed at intervals of 1 minute, and shaking was performed every 10 minutes during the reaction. The pathogen control group used hard water without sterilant diluent, and the toxic control group used light water diluted disinfectant diluent.

Immediately after the reaction was completed, 1 mL was taken out and mixed in 5 mL of neutralization medium (containing 10% (v / v) of fetal bovine serum immobilized on α-MEM containing antibiotic and antimycotic) Then, the neutralization reaction solution was added to a viral medium containing 0.3% (v / v) tryptose phosphate boroth, 0.02% (v / v) yeast extract and 2 μg / mL trypsin in a viral medium containing antibiotic antimycotic The diluted solution was diluted to a concentration of 10 ^-1 , 10 ^-2 , 10 ^-3 , 10 ^-4 and 10 ^{-5 using} a stock solution of vero cell (80-90%) with 8 wells per dilution Respectively.

After inoculation, the cells were incubated at 37 ° C for 5 days. CPE formation was observed daily, and the virus content was calculated by the Karber method.

The pathogen control was tested in the light water condition without the disinfectant and the neutralization reaction step confirmed that the activity of the pathogen was more than 2 × 10 ⁵ TCID ₅₀ / mL. In the toxic control group, it was confirmed that no cytotoxicity was caused by the disinfectant disinfectant.

4-4. Porcine respiratory and reproductive syndrome virus (PRRS)

Prior to the test, 200 μL of each well of a 96-well plate was plated at 2 × 10 ⁵ cells / mL in a Porcine Alveolar Macrophage cell (using RPMI-1640 containing antibiotic / antimycotic as a culture medium) ₂ < / RTI > conditions.

The sterilization disinfectants of Examples 1 to 2 and Comparative Examples were immersed in hard water (containing 0.305 g of CaCl ₂ and 0.139 g (w / v) of MgCl ₂ .6H ₂ O in 1 L of distilled water) and an organic material (fetal calf serum 5 % (v / v)) as follows. The disinfectant of Example 1 was diluted with water at 10,000, 16,000 and 20,000 magnifications, and the organic matter was diluted at a magnification of 2,000, 2,400 and 3,000. The disinfectant of Example 2 was diluted with water at 10,000, 16,000, and 20,000 magnifications and diluted with an organic material at 2,000, 2,400, and 3,000 magnifications. In addition, the sterilization disinfectant of the comparative example was diluted with water at 10,000, 16,000, and 20,000 magnifications, and diluted with organic matter at 1,600, 2,000, and 2,400 magnifications.

Thereafter, 1.0 mL (1 × 10 1 to 1 × 10 ⁷ EID ₅₀ / mL) of virus solution stored at 4 ° C. was added to 19 mL of hard water or organic matter, which is a virus dilution solution, and mixed. 2.5 mL of the virus solution was added to each test tube containing the same amount of the disinfectant solution of Examples 1 and 2 and the comparative example, respectively (5 mL in total), and reacted at 4 ° C for exactly 30 minutes. At this time, each test tube treatment was performed at intervals of 1 minute, and shaking was performed every 10 minutes during the reaction. The pathogen control group used hard water without sterilant diluent, and the toxic control group used light water diluted disinfectant diluent.

Immediately after the reaction was completed, 1 mL was taken out and mixed in 5 mL of neutralization medium (containing 10% (v / v) fetal bovine serum unmixed with RPMI-1640 containing antibiotic antimycotic) at 37 ° C in advance. Then, the neutralization reaction solution was diluted with a virus medium (RPMI-1640 containing antibiotic / antimycotic) at a concentration of 10 ^-1 , 10 ^-2 , 10 ^-3 , 10 ^-4 and 10 ^-5 Diluted and 100 μL each were inoculated into MARC-145 cells in which 8-well monolayer (80-90%) was formed per dilution magnification.

After inoculation, the cells were incubated at 37 ° C for 5 days. CPE formation was observed daily, and the virus content was calculated by the Karber method.

The pathogen control was tested in the light water condition without the disinfectant and the neutralization reaction step confirmed that the activity of the pathogen was more than 2 × 10 ⁵ TCID ₅₀ / mL. In the toxic control group, it was confirmed that no cytotoxicity was caused by the disinfectant disinfectant.

4-5. Foot and mouth disease virus ( Sterilization disinfection power for FMD)

The test is carried out before Lamb kidney cell (L-glutamine, sodium bicarbonate, non-essential amino acid, sodium pyruvate, FBS and antibiotics using the addition of EMEM in the culture medium) to ^{3 × 10 5 cell / mL 6} well so that the , and cultured in a 5% CO ₂ incubator at 37 ° C. The FMD virus, which was confirmed to have a virus titer of 10 ^7.0 TCID ₅₀ / mL or higher during the subculture, was stored at 4 ° C for a short time until immediately before use.

The sterilization disinfectants of Examples 1 to 2 and Comparative Examples were prepared by using an organic substance-containing water (prepared by adding 0.304 g of CaCl ₂ and 0.139 g (w / v) of MgCl ₂ .6H ₂ O to 1 L of distilled water and containing 1% of FBS) And diluted as follows. The sterilization disinfectants of Examples 1, 2 and Comparative Example were diluted at a magnification of 100, 200, 300, 600 and 900, respectively.

Thereafter, 1.8 mL of the virus solution at 4 ° C (1 × 10 ⁵ to 1 × 10 ⁷ TCID ₅₀ / mL) was added to 0.2 mL of a disinfectant diluted solution 10 times more concentrated than the final dilution, mixed with the disinfectant diluent and virus solution mixture 4 Lt; 0 > C for 30 minutes. The test mixture was thoroughly mixed with the virus and disinfectant immediately before the reaction was completed.

To neutralize the disinfectant immediately after the reaction, 0.05 mL of the test mixture was added to 5 mL of neutralization medium (EMEM containing 2% antibiotic and 5% FBS) stored in a 4 ° C water bath. The final mixture was diluted 10-fold (10 ^-2 to 10 ^-6 ) 10 times with 4% neutralization medium. 0.2 mL of each final dilution, including the negative control (inoculated with distilled water) and the positive control (1% NaOH), was inoculated into a 96-well plate with a cell monolayer.

And incubated for 1 hour at 37 ° C in a 5% CO ₂ humidified incubator, shaking well every 8 to 10 minutes. After 1 hour of inoculation, 2.5 mL of a medium containing 1% methylcellulose was added, followed by further incubation for 2 days in a 5% CO ₂ humidified incubator at 37 ° C.

After incubation, the cells were washed with 0.01-0. 5% citric acid and stained with amidoblack. The number of plaques was counted to calculate the PFU value.

4-6. Experiment result

The results of the sterilization test on 4-1 to 4-5 were compared with the pathogen control to confirm the effective dilution factor. That is, the dilution factor was determined as the effective dilution factor, which was confirmed to decrease by at least 10 log ⁴ (at least 4 log10) compared to the pathogen control. The median of the results within an error range of 20% was determined as the final effective dilution factor for the disclosed product three times. The results are shown in Table 4 below.

As shown in Table 4 above, the chlorine-based disinfectant of the present invention is effective against avian influenza viruses ( Avian influenza virus , AI), Newcastle disease ( Newcastle disease virus, ND), porcine epidemic diarrhea virus (Porcine epidemic diarrhea virus, PED) , porcine respiratory and reproductive syndrome virus (Porcine respiratory and reproductive syndrome virus, PRRS), foot-and-mouth disease virus (Foot and mouth disease virus, FMD ) such as viruses And showed an effective disinfecting power at a dilution ratio similar to that of Comparative Example 1, even when there was no chlorine odor.

Although the present invention has been described in terms of the preferred embodiments mentioned above, it is possible to make various modifications and variations without departing from the spirit and scope of the invention. It is also to be understood that the appended claims are intended to cover such modifications and changes as fall within the scope of the invention.

Claims (5)

Sodium dicarboxylate isocyanurate; And
Wherein the chlorine-free disinfecting composition comprises any one selected from the group consisting of sulfamic acid or a mixture of sodium peroxide and sulfamic acid. The method according to claim 1,
The chlorine-based disinfecting composition may be selected from the group consisting of Salmonella typhimurium , Brucella ovis , Clostridium perfringens , Avian influenza virus (AI), Newcastle disease virus , ND), porcine epidemic diarrhea virus (Porcine epidemic diarrhea virus, PED) , swine respiratory and reproductive syndrome virus (Porcine respiratory and reproductive syndrome virus, PRRS) and foot-and-mouth disease virus (least any one selected from Foot and mouth disease virus, FMD) Wherein the chlorine-free sterilizing agent is used for sterilization or disinfection. delete The method according to claim 1,
50 to 80 parts by weight of sodium dichloroisocyanurate sodium based on 100 parts by weight of the total composition; And 10 to 50 parts by weight of one selected from the group consisting of sulfamic acid or a mixture of sodium peroxide and sulfamic acid. The method according to claim 1,
The composition may further comprise at least one selected from the group consisting of ascorbic acid, hydrogen peroxide, sodium bicarbonate, sodium carbonate, and citric acid. A chlorine-based disinfecting composition comprising chlorine removed.