JP2002003317A - Bactericidal composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a bactericide having a high safety, gentle to environment and exhibiting a high bactericidal potency. SOLUTION: This bactericidal composition provides a bactericidal liquid exhibiting 10-2000 ppm iodide concentration, 1-100 ppm hydrogen peroxide concentration, 0.01-2 U/ml soybean peroxidase activity (measured by a pyrogallol method) and 5.0-10.0 molar ratio of the iodide to hydrogen peroxide on dissolving it in an aqueous medium.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an environmental disinfectant for hospitals, food factories, livestock pens, etc. and a disinfectant for medical equipment, food processing equipment, cooking utensils, tableware, fingers and the like.

[0002]

2. Description of the Related Art Environmental disinfectants for hospitals, food factories, livestock barns, etc.
Various disinfectants have been developed as disinfectants for medical equipment, food processing equipment, cooking utensils, tableware, fingers, etc., alcohol-based, aldehyde-based, chlorine-based, quaternary ammonium-based, amphoteric surfactant-based, Biguanide bactericides can be exemplified. Alcohol-based fungicides have disadvantages such as being ineffective against bacterial spores, having poor residual effects due to volatilization, and having reduced efficacy in the presence of organic substances. There are problems such as a weak effect on spores and mold. Chlorine disinfectants have a weak effect on Mycobacterium tuberculosis and bacterial spores. Is generated. Quaternary ammonium fungicides are
There are problems such as ineffectiveness against Mycobacterium tuberculosis and bacterial spores, weak effect on mold and virus, and decrease in efficacy in the presence of organic matter. Amphoteric surfactant fungicides have no effect on bacterial spores and have a weak effect on Mycobacterium tuberculosis, viruses and mold. The biguanide fungicides have problems such as being ineffective against acid-fast bacteria such as Mycobacterium tuberculosis, bacterial spores and viruses, producing resistant bacteria, and having different efficacy depending on strains. Until now, no fungicide which is excellent in safety to the human body and the environment and has a strong effect on a wide range of microorganisms such as bacteria, molds, viruses and protozoa has not been found.

[0003]

SUMMARY OF THE INVENTION An object of the present invention is to solve the above-mentioned problems in the prior art and to provide a fungicide which is safe, environmentally friendly and has a strong bactericidal activity.

[0004]

Means for Solving the Problems The inventors of the present invention have conducted intensive studies in view of the above problems, and as a result, have found that a highly safe iodide,
By mixing peroxide and peroxidase at the right concentration and hydrogen peroxide and iodide in the right molar ratio,
The present inventors have found that remarkable bactericidal activity against a wide range of microorganisms is developed, and completed the present invention. That is, the present invention provides an iodide concentration of 10 ppm to 2000 ppm when dissolved in an aqueous medium,
The hydrogen peroxide concentration is 1 ppm to 100 ppm, the peroxidase activity is 0.01 U / ml to 2 U / ml (measured by pyrogallol method), and the molar ratio of iodide to hydrogen peroxide is 5.0 to
The present invention relates to a disinfectant composition for supplying a disinfectant solution which is 10.0.

[0005]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention provides a fungicide composition containing iodide, peroxide and peroxidase as essential components. Hereinafter, the present invention will be described in detail. The fungicidal composition of the present invention contains iodide, peroxide, and peroxidase as components. In addition to these, it is also possible to add a pH adjuster, a peroxidase enhancer, a peroxidase stabilizer and the like. As iodide, potassium iodide,
Although sodium iodide is suitable, any compound that generates iodide ions in an aqueous medium may be used, and is not limited thereto. The processing concentration range of iodide is 10ppm ~
2000 ppm, but the preferred concentration range is 500 ppm to 1000 pp
m. As the peroxide, hydrogen peroxide, sodium percarbonate and sodium perborate are suitable, but any compound that generates hydrogen peroxide in an aqueous medium may be used, such as sodium peroxide, methyl peroxide, and peroxide. Ethyl, peracetic acid,
Peroxosulfuric acid, peroxophosphoric acid and the like can also be used. Peroxide has a hydrogen peroxide concentration in an aqueous medium of 1 ppm or more.
It is added so as to be 100 ppm, preferably 10 ppm to 50 ppm. Iodide and hydrogen peroxide are fed into the aqueous medium such that the molar ratio of iodide to hydrogen peroxide is between 5.0 and 10.0.

[0006] Examples of peroxidases include plant-derived peroxidases such as horseradish and rice, and as peroxidases derived from filamentous fungi, incomplete fungi such as Arthromyces ramosus and Geotrichum candidu.
m, Curvularia inaequalis, Drechsiera biseptata, U
locladium botrytis, Pellicularia filamentosa, basidiomycete Coprinus cinereus peroxidase, white rot fungus Phanerochaete chrysosporium, Bjerkandera adust
a, Trametes vesicolor, Pleurotus ostreatus, Phlebi
Lignin peroxidase and manganese peroxidase such as a radiata are exemplified. Bacterial peroxidases include Flavobacterium sp., Bacillus stearo
Examples are peroxidases of thermophilus and Streptomyces viridosporus. However, the peroxidase that can be used in the present invention is not limited to these. Peroxidase can also be used after being chemically modified and immobilized. Regarding chemical modification, polyethylene glycol, methoxypolyethylene glycol, polyethylene glycol-maleic anhydride copolymer, polyoxyalkylene derivative-maleic anhydride copolymer, isobutylene-maleic anhydride copolymer, methoxypolyethylene glycol as a chemical modifier -Succinimidyl succinate copolymer, acetic anhydride and the like are exemplified, but not limited thereto.

[0007] With respect to immobilization of peroxidase, examples of the immobilization carrier used in the carrier binding method include cellulose derivatives such as carboxymethylcellulose and diethylaminoethylcellulose, agarose, dextrin, chitin, chitosan, polyacrylamide gel, silica gel, and the like.
Examples thereof include, but are not limited to, ion exchange resins such as alumina, celite, latex, porous glass, magnetic particles, Amberlite, Diaion, DEAE-Sephadex, and DEAE-cellulose. Examples of the immobilization carrier used in the entrapping method include collagen, agarose, alginic acid, carrageenan, polyacrylamide, various photocrosslinkable resins,
Examples include various photosensitive resins, collodion, cellulose nitrate, divinylbenzene polymer, and phospholipid, but are not limited thereto. Examples of the cross-linking agent used in the cross-linking method include a maleimide derivative, glutaraldehyde, and hexamethylene diisothiocyanate, but are not limited thereto. Peroxidase is immobilized on the above-mentioned immobilization carrier or the like by a known method.
Peroxidase is added to the aqueous medium so as to be 0.01 U / ml to 2 U / ml depending on the conditions of use and the type of peroxidase.

[0008] Examples of peroxidase enhancers include 10-methylphenothiazine and p-hydroxybenzenesulfonic acid, but they are not limited to these as long as they enhance peroxidase activity. In addition, as a stabilizer for peroxidase,
Examples include, but are not limited to, potassium chloride, phenol, hematin, sucrose, glycerin, and ethanol. Each component is stored in such a manner that the reaction does not proceed until it requires bactericidal activity, and at the time of use, each component is mixed in an aqueous medium such that each has an appropriate concentration and molar ratio. This can be done by storing each component separately or by separating iodide and peroxidase from peroxide. In addition, each component is granulated, encapsulated,
It is also possible to store in the form of a mixture in which the components are mixed so that they do not react until they are mixed in an aqueous medium. Examples of excipients used for granulation include anhydrous sodium sulfate,
Dextrin, glucose, lactose and the like can be exemplified. The granules can be protected with a water-soluble polymer film or the like, or surface-coated with hydroxypropylmethylcellulose or the like.

[0009]

EXAMPLES The present invention will be described specifically with reference to Examples.
The present invention is not limited to the following examples. Example 1 Escherichia coli (Esche) was placed in a test tube containing 10 ml of ordinary broth medium.
richia coli), Staphylococcus aureu
s), inoculated with one platinum loop of Pseudomonas aeruginosa or Salmonella enteritidis, 37 ° C
And shaking culture was performed until the OD ₆₁₀ reached about 1.0. It was diluted 10-fold with 50 mM phosphate buffer (pH 6.0) to obtain a test bacterial solution. Potassium iodide 16% aqueous solution, hydrogen peroxide 0.6
A 0.01% aqueous solution and a horseradish peroxidase 200 U / ml aqueous solution were added in 0.01 ml portions to 1.97 ml of the above phosphate buffer.
After mixing with 0.1 ml of the test bacterial solution, leave at 30 ° C for 0.5 min.
After 5 minutes, 5 minutes, 15 minutes, 30 minutes and 60 minutes, the treatment liquid was sampled, diluted appropriately, and spread on a normal agar medium to measure the number of viable cells. Bacteria were similarly inoculated into a comparative solution having a predetermined concentration, and the number of viable bacteria was measured over time. The time at which the viable cell count fell below the detection limit (5 / ml) was taken as the sterilization time. Table 1 shows the results. The germicidal solution of the present invention was sterilized against all the bacteria in 0.5 minutes and showed the strongest efficacy.

Table 1 Sterilization time (min) Escherichia coli Staphylococcus aureus Pseudomonas aeruginosa Salmonella Bacterial solution of the present invention 0.5 0.5 0.5 0.5 Comparative solution 1 0.5 11 1 Comparative solution 2 0.5 1 1 0.5 Number of initial bacteria: about 10 ⁶ / ml Comparative liquid 1: sodium hypochlorite (effective chlorine concentration 150 ppm) Comparative liquid 2: iodohole (effective iodine concentration 100 ppm)

Example 2 Escherichia coli (Esche) was placed in a test tube containing 10 ml of ordinary broth medium.
richia coli), Staphylococcus aureu
s), one platinum loop of Pseudomonas aeruginosa or Salmonella enteritidis was inoculated at 37 ° C.
And shaking culture was performed until the OD ₆₁₀ reached about 1.0. It was diluted 5-fold with 50 mM phosphate buffer (pH 6.0) to give a test bacterial solution. 800mg of potassium iodide, sodium percarbonate
10 mg powder of horseradish peroxidase 50 U
It was dissolved in 100 ml of 50 mM phosphate buffer (pH 6.0). After mixing with 2 ml of this bactericidal solution and 0.05 ml of the test bacterial solution, leave at 30 ° C.
After 1 minute, 1 minute, 5 minutes, 15 minutes, 30 minutes, and 60 minutes, the treatment liquid was sampled, diluted appropriately, spread on a normal agar medium, and the viable cell count was measured. Inoculate the bacteria in the same manner to the comparative solution of the predetermined concentration,
The number of viable bacteria was measured over time. The time at which the viable cell count fell below the detection limit (5 / ml) was taken as the sterilization time. Table 2 shows the results.
The germicidal solution of the present invention sterilized all the bacteria in 0.5 minutes and showed the strongest efficacy.

Table 2 Sterilization time (min) E. coli Staphylococcus aureus Pseudomonas aeruginosa Salmonella fungicide of the present invention 0.5 0.5 0.5 0.5 Comparative solution 1 0.5 11 1 Comparative solution 2 0.5 1 1 0.5 Number of initial bacteria: about 10 ⁶ / ml Comparative liquid 1: sodium hypochlorite (effective chlorine concentration 150 ppm) Comparative liquid 2: iodohole (effective iodine concentration 100 ppm)

Example 3 Black mold Aspergil was added to potato dextrose agar slant medium.
lus niger was inoculated and cultured at 25 ° C for 1 week. 50 for this
A spore suspension was prepared by adding an mM phosphate buffer (pH 6.0) and stirring. This was filtered through gauze to remove mycelial fragments, and used as a test spore solution. Potassium iodide 16%
An aqueous solution, a 0.6% aqueous solution of hydrogen peroxide, and a 200 U / ml aqueous solution of horseradish peroxidase were added in 0.01 ml portions to 1.97 ml of the above phosphate buffer. After mixing with 0.1 ml of the test bacterial solution, leave at 30 ° C, sample the treated solution after 0.5, 1, 5, 15, 30, and 60 minutes, dilute appropriately, and place on a potato dextrose agar medium. The number of viable spores after coating was measured. Spores were similarly inoculated into a comparative solution having a predetermined concentration, and the number of viable spores was measured over time. The time when the number of surviving spores fell below the detection limit (5 / ml) was defined as the sterilization time. Table 3 shows the results. The germicidal solution of the present invention sterilized black mold in 0.5 minutes and showed an effect more than that of the comparative solution.

[0014] Initial spore count: about 10 ⁶ / ml Comparative solution 1: sodium hypochlorite (effective chlorine concentration 150pp
m) Comparative solution 2: iodine hole (effective iodine concentration 100ppm)

Example 4 Bacillus subtilis and Bacillus cereus were inoculated on a normal agar slant medium, and cultured at 30 ° C. for 7 days to form spores. Add it to a 50 mM phosphate buffer (p
H6.0), and killed the vegetative cells by heat treatment (100 ° C, 5 minutes) to obtain a test bacterial solution. Potassium iodide
A 16% aqueous solution, a 0.6% hydrogen peroxide aqueous solution, and a horseradish peroxidase 200 U / ml aqueous solution were added in 0.01 ml portions to 1.97 ml of the above phosphate buffer. After mixing with 0.5 ml of test bacterial solution, 30
After standing at 0.5 ° C., the treated solution was sampled 0.5 minutes, 1 minute, 5 minutes, 15 minutes, 30 minutes, and 60 minutes, diluted appropriately, spread on a normal agar medium, and counted the number of viable spores. As a comparison, bacteria were similarly inoculated into a composition solution containing 800 ppm of potassium iodide, a concentration of hydrogen peroxide of 3 ppm, and 1 U / ml of horseradish peroxidase (comparative composition solution), and a comparative agent of a predetermined concentration. The number was measured. The time when the number of surviving spores was below the detection limit (5 / ml) was defined as the sterilization time. Table 4 shows the results. The germicidal solution of the present invention showed more efficacy against the spores of any bacteria than the comparative composition solution and the comparative solution.

Table 4 Sterilization time (min) Bacillus subtilis B. cereus fungicide of the present invention 5 0.5 Comparative solution 60 60 Comparative solution 1>60> 60 Comparative solution 2 60 1 Number of first spores: about 10 ⁶ / ml Comparative solution 1: Sodium hypochlorite (effective chlorine concentration 150 ppm) Comparative solution 2: iodohole (effective iodine concentration 100 ppm)

Example 5 A 16% aqueous solution of potassium iodide, a 0.6% aqueous solution of hydrogen peroxide and a 200 U / ml aqueous solution of horseradish peroxidase were added in 0.1 ml portions to 19.7 ml of a 50 mM phosphate buffer (pH 6.0). Echo virus solution or poliovirus solution (about 4000TC
(D50 / ml) After mixing with 5 ml, leave at 30 ° C for 0.5 minutes, 1 minute, 5 minutes.
After minutes, 15, 30, and 60 minutes, the active ingredient was inactivated with 0.1N sodium thiosulfate. 0.1 ml of the treatment solution was sampled and added to five test tubes in which HeLa cells were cultured. 6
The observation was continued for one day, and the presence or absence of inhibition of CPE (cytopathic effect) was examined. When CPE inhibition was observed in all test tubes, virus inactivation was determined to have been observed. Table 5 shows the results. The germicidal solution of the present invention showed a strong inactivating activity against the tested virus.

Table 5 Inactivation time of virus sterilization (min) Echovirus Poliovirus Disinfectant of the present invention 1 0.5 Comparative solution 11 Comparative solution: sodium hypochlorite (effective chlorine concentration 150 ppm)

[0019]

According to the present invention, there is provided a disinfectant which is safe, environmentally friendly and has a strong disinfecting power.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61K 33/40 A61K 33/40 38/44 A61P 31/04 A61P 31/04 A61K 37/50

Claims (1)

[Claims] When dissolved in an aqueous medium, an iodide concentration is 10 ppm to 2000 ppm, and a hydrogen peroxide concentration is 1 ppm to 100 pp.
m, a disinfectant characterized by supplying a disinfectant having a peroxidase activity of 0.01 U / ml to 2 U / ml (measured by the pyrogallol method) and a molar ratio of iodide to hydrogen peroxide of 5.0 to 10.0. Composition.