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WO2017017810A1 - Method for killing spores - Google Patents

Method for killing spores

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Publication number
WO2017017810A1
WO2017017810A1 PCT/JP2015/071499 JP2015071499W WO2017017810A1 WO 2017017810 A1 WO2017017810 A1 WO 2017017810A1 JP 2015071499 W JP2015071499 W JP 2015071499W WO 2017017810 A1 WO2017017810 A1 WO 2017017810A1
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WO
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Patent type
Prior art keywords
preferably
step
less
acid
minutes
Prior art date
Application number
PCT/JP2015/071499
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French (fr)
Japanese (ja)
Inventor
知美 阪井
Original Assignee
花王株式会社
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES, AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N33/00Biocides, pest repellants or attractants, or plant growth regulators containing organic nitrogen compounds
    • A01N33/02Amines; Quaternary ammonium compounds
    • A01N33/12Quaternary ammonium compounds
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES, AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N43/00Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/34Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom
    • A01N43/40Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom six-membered rings
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION, OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS, OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS, OR SURGICAL ARTICLES
    • A61L2/00Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
    • A61L2/16Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using chemical substances
    • A61L2/18Liquid substances or solutions comprising solids or dissolved gases
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61LMETHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION, OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS, OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS, OR SURGICAL ARTICLES
    • A61L2/00Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
    • A61L2/16Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using chemical substances
    • A61L2/23Solid substances, e.g. granules, powders, blocks, tablets

Abstract

A method for killing spores according to the present invention comprises carrying out step 1, step 2 and step 3. Specifically, step 1 comprises bringing a spore-forming bacterium into contact with dipicolinic acid or a salt thereof, step 2 comprises bringing the spore-forming bacterium into contact with a cationic surfactant, and step 3 comprises warming the spore-forming bacterium to 50ºC or higher. There may be a period during which step 1, step 2 and step 3 are carried out simultaneously.

Description

Quit method

The present invention relates to a stop process of spore-forming bacteria.

Spore-forming bacteria such as Bacillus or Clostridium genus, making a strong shell structure to form spores with extremely high resistance such as heat or agents. Certain spore-forming bacteria are known to produce toxins and enters the human body. For example, in the medical field, but the linen items such as sheets and pillowcases are mainly heated disinfected, can not be sterilized spore-forming bacteria is a heat-resistant at heating sterilization. For this reason, also it has occurred damage out to the dead in the hospital infection of spore-forming bacteria through the linen products.

To sterilize the spore-forming bacteria, often or sterilized with high pressure steam, strong chemical disinfectants such as hypochlorite Na is or used in high concentrations. In the food processing, severe heat treatment as spores measure or cold distribution methods have been taken.

The US2014 / 0308162 discloses, disinfecting and rinsing composition used in the laundry detergent are disclosed. Further, Japanese Patent Publication No. US2014 / 0238445, phosphino comprises a composition disinfecting and rinsing steps using, including a washing step and percarboxylic acid with a composition comprising succinic acid, washed, the method of disinfecting and rinsing is disclosed ing. More WO2013 / 079308 discloses discloses a to sterilize bacteria including spores. US2004 / 0058878 discloses are disclosed for improving the stop effect by combining germinating agent and quaternary ammonium salts.

The present invention comprises the steps 1 below, step 2, and stop method for performing related processes 3.
Step 1: a step of previously contacting the spore-forming bacteria and dipicolinic acid or a salt thereof;
Step 2: Step left in contact with the spore-forming bacteria and a cationic surfactant; and Step 3: a step of heating to more than 50 ° C. The spore-forming bacteria.
(However, and it ends the process 3 after starting the steps 1 and 2).

The present invention contains dipicolinic acid or a salt thereof and a cationic surfactant, about Yamesuke composition.

Detailed Description of the Invention

In the previous technique disclosed spores, i.e. not have sufficient bactericidal effect against spore-forming bacteria dormant, the effect of reducing the number of spores was insufficient.

Stop method by high concentration of sterilant or high-pressure steam sterilization is not suitable for disinfecting a wide range of environments, such as hospitals, since it is possible that there is toxicity to equipment damage resistance and the human body, Ya intended use there is a restriction on the use environment.

In food processing, the severe heat treatment as spores measure, or has a low temperature distribution methods have been taken, the deterioration of the flavor and quality, also issues such as power cost occurs.

The present invention is a spore-forming bacteria as a harm in the medical and food industry, to efficiently and stop method capable of effectively sterilized. Furthermore, the present invention relates to quit aid composition used to sterilize the spore-forming bacteria. Primarily intended to kill germinated bacteria state with spores and "stop" in the present invention.

The present inventor has been made in view of the above problems, intensive studies were carried out results, and dipicolinic acid (2,6 pyridinedicarboxylic acid) or a salt thereof and a cationic surfactant by keeping in contact with the spore-forming bacteria, gentle effectively it found that can be stopped by heating, leading to the present invention.

According to the present invention, the spore-forming bacteria, quit method can be sterilized is provided. Furthermore, in the present invention, it stops aid composition used to sterilize the spore-forming bacteria is provided.

The spore-forming bacteria, by forming the spores in nutrient absence refers to bacteria resistant to certain heat treatment or drying. The spores, bacteria form, showed a strong shell structure that bacteria forms, distinguished from the bacteria itself. The killing bud subject of the present invention, the "spore-forming fungi", medical facilities and food, is a common spore-forming bacteria present in the beverage product. For example, Bacillus cereus (Bacillus cereus) and Bacillus subtilis (Bacillus subtilis), such as Bacillus (Bacillus) belonging to the genus of bacteria, Clostridium difficile (Clostridium difficile), such as Clostridium (Clostridium) bacteria of the genus, Amphitheater Bacillus (Amphibacillus) bacteria of the genus , Sporosarcina (Sporosarcina) genus bacteria, Geobacillus (Geobacillus) genus bacteria, Airy Bacillus (Aeribacillus) a bacterium of the genus, and the like Alicyclobacillus (Alicyclobacillus) genus of bacteria. When these spore-forming bacteria form spores, spore-forming bacteria have heat resistance. Here, the heat resistance, as indicated by spore-forming bacteria were subjected to an embodiment of the present invention, while forming the initial bacteria number 10 7 ~ 10 9 CFU / mL of spores, 30 min at 80 ° C. or when warmed, 10 7 CFU / mL or more spore-forming bacteria means that can survive.

Primarily intended to kill germinated bacteria state with spores and "stop" in the present invention. Germination and is that spores spore-forming bacteria germinate, thereby pointing normal growth, a phenomenon in which the cells having a metabolic capacity. The stop also include reducing the number of bacteria whole bacteria with spores present in the population. The stop effect is not particularly limited, for example, as shown in the examples, the viable cell count of bacteria having spore, those initial number of bacteria was 10 8 CFU / mL is 10 7 CFU / mL showing a state in which less than.

The present invention includes the steps 1 below, step 2, and by performing step 3, has an excellent stop effect.
Step 1: a step of previously contacting the spore-forming bacteria and dipicolinic acid or a salt thereof;
Step 2: Step left in contact with the spore-forming bacteria and a cationic surfactant; and Step 3: a step of heating to more than 50 ° C. The spore-forming bacteria.
(However, and it ends the process 3 after starting the steps 1 and 2).

In the present invention, the order of steps 1, 2 and 3 is not particularly limited. However, and it ends the process 3 after starting the steps 1 and 2. That is, Step 1, Step 2, and in a series of operations including step 3, is not included to initiate either after completion of Step 3 of the process 1 or process 2, or both.

Reason for having a stop effect quit method is superior to the present invention is not clear is considered as follows. Step 1, by performing the steps 2 and 3, spores germinate. The cationic surfactant, increased affinity of dipicolinic acid or a salt thereof and spore-forming bacteria, easy dipicolinic acid or a salt thereof enters sporulation the bacteria effectively germination is presumed to be promoted when heating that. Germinated spore-forming bacteria will be readily sterilizable in step 2 and step 3.

Moreover, the action of dipicolinic acid or spore formation in bacteria salt thereof is not a working through a specific receptor-specific by species, believed to have quit effect regardless of the type of spore-forming bacteria.

Note that, by confirming the reduction of the heat resistance of the bacteria, it is possible to confirm that germinated. Decrease in heat resistance, the spore-forming bacteria, for example, can be treated under 80 ° C. environment by examining the number of bacteria surviving to confirm.

In the present invention, when using a dipicolinic acid salt, as a counter ion of the dipicolinic acid is not particularly limited, alkali metal ions such as sodium ions and potassium ions; alkaline earth metal ions such as calcium ions and magnesium ions are exemplified that.

In the present invention, the term "cationic surfactant" refers to surfactant hydrophilic portion is charged cation when dissolved in solution, but are not limited to, primary ammonium salts, secondary ammonium salt tertiary ammonium salts, quaternary ammonium salts and the like. Cationic surfactants of the present invention, more specifically, alkyl trimethylammonium salts, alkyl triethyl ammonium salts, alkyl dimethyl ethyl ammonium salt, alkyl methyl diethyl ammonium salt, dialkyl dimethyl ammonium salts, dialkyl diethyl ammonium salts, dialkyl ethylmethyl ammonium salts, benzalkonium salts, alkyl pyridinium salts, quaternary ammonium salts such as alkyl benzethonium salts, and primary ammonium salts such as alkyl amine salts.

Here, alkyltrimethylammonium salts, alkyl triethyl ammonium salts, alkyl dimethyl ethyl ammonium salts and alkyl methyl diethyl ammonium salt refers to a compound represented by the following general formula (1).

Figure JPOXMLDOC01-appb-C000002

Wherein any one of R 11 ~ R 14, hydroxyl group, an ester group, an amide group hydrocarbon group having 3 or more carbon atoms which may have a remaining three of R 11 ~ R 14 represents a methyl group or an ethyl group, X - represents an inorganic or organic anionic compounds. ]

Here, the carbon number of the hydrocarbon group having 3 or more carbon atoms, from the viewpoint of further improving the stop effect, preferably 6 or more, more preferably 10 or more, more preferably 12 or more, more preferably is 14 or more, and, preferably 22 or less, more preferably 20 or less, more preferably 18 or less. Of 3 or more hydrocarbon group having a carbon may be an alkyl or alkenyl group linear or branched, preferably a straight chain alkyl group. X - is chloride ion, a halogen ion such as bromide ion, sulfate ion, phosphate ion, hydrogen phosphate ion, dihydrogen phosphate ion, nitrate ion, carbonate ion, bicarbonate ion, acetate ion, and the like. Preferably an inorganic ion, more preferably a halogen ion, more preferably a chloride ion.

Here, dialkyl dimethyl ammonium, dialkyl diethyl ammonium salts and dialkyl methyl ammonium salt, refers to a compound represented by the following general formula (2).

Figure JPOXMLDOC01-appb-C000003

Wherein any two of R 21 ~ R 24, hydroxyl group, an ester group, which may have an amide group may of 3 or more carbon atoms hydrocarbon group, the remaining two of R 21 ~ R 24 represents a methyl group or an ethyl group, X - represents an inorganic or organic anionic compounds. Here, any two of the hydroxyl groups of R 21 ~ R 24, an ester group, a hydrocarbon group of good 3 or more carbon atoms have an amide group may not be the same. ]

Here, the carbon number of 3 or more hydrocarbon group having a carbon, from the viewpoint of further improving the stop effect, preferably 6 or more, more preferably 10 or more, and preferably 22 or less, more preferably 20 or less, more preferably 18 or less, more preferably 14 or less. Of 3 or more hydrocarbon group having a carbon may be an alkyl or alkenyl group linear or branched, preferably a straight chain alkyl group. X - is chloride ion, a halogen ion such as bromide ion, sulfate ion, phosphate ion, hydrogen phosphate ion, dihydrogen phosphate ion, nitrate ion, carbonate ion, bicarbonate ion, acetate ion, and the like. Preferably an inorganic ion, more preferably a halogen ion, more preferably a chloride ion.

Benzalkonium salts, refers to a compound represented by the following general formula (3).

Figure JPOXMLDOC01-appb-C000004

Wherein, R 31 is a hydrocarbon group, X - represents an inorganic or organic anionic compounds. ]

The number of carbon atoms in R 31 is preferably 6 or more, more preferably 10 or more, more preferably 12 or more, more preferably 14 or more, and preferably 22 or less, more preferably 20 or less, more preferably 18 or less. R 31 may be an alkyl or alkenyl group linear or branched, preferably a straight chain alkyl group. X - is preferably chloride ion, a halogen ion such as bromide ion, sulfate ion, include phosphate, hydrogen phosphate ions, dihydrogen phosphate ions, nitrate ions, carbonate ions, bicarbonate ions, acetate ions or the like It is. Preferably an inorganic ion, more preferably a halogen ion, more preferably a chloride ion.

Wherein alkyl pyridinium salts, alkyl benzethonium salt or "alkyl" as referred alkylamine salts, preferably refers to the number 3 or more hydrocarbon group having a carbon. The number of carbon atoms of the hydrocarbon group having 3 or more carbon atoms is preferably 6 or more, more preferably 10 or more, and preferably 22 or less, more preferably 20 or less, more preferably 18 or less. Of 3 or more hydrocarbon group having a carbon may be an alkyl or alkenyl group linear or branched, preferably a straight chain alkyl group.

The cationic surfactants include dialkyl dimethyl ammonium salts, alkyl trimethyl ammonium salt or benzalkonium salt, more preferably a dialkyl dimethyl ammonium salt and alkyltrimethylammonium salt, more preferably an alkyl trimethyl ammonium salt.

Here, salts include, but are not limited to, chloride, halides of bromide such as, sulfate, phosphate, hydrogen phosphate, dihydrogen phosphate, nitrate, carbonate, bicarbonate, acetate, etc. and the like. Preferably an inorganic salt, more preferably a halide, more preferably chloride.

The cationic surfactants include dialkyl dimethyl ammonium chloride, preferably an alkyl trimethyl ammonium chloride or benzalkonium chloride, more preferably dialkyldimethylammonium and alkyltrimethylammonium chloride chloride, alkyl trimethyl ammonium chloride is more preferred.

Molar ratio cationic surfactants dipicolinic acid ([molar concentration of dipicolinic acid] / [molar concentration of cationic surfactant]) is preferably 1/1000 or more, more preferably 1/100 or more, more preferably 1/50 or more, more preferably 1/20 or more, more preferably 1/10 or more, more preferably 1/5 or more, more preferably 1/2 or more, and, preferably 1,000 or less, more preferably 100 or less, more preferably 50 or less, more preferably 20 or less, more preferably 10 or less, more preferably 5 or less, more preferably 2 or less.

<Steps 1 and 2>

Step 1 of the present invention is a process to keep contacting the spore-forming bacteria and dipicolinic acid or a salt thereof. Step 2 of the present invention is a process to keep contacting the spore-forming bacteria and a cationic surfactant. Here, the steps 1 and 2, includes any of the following aspects.
(1) aspect of performing steps 1 and 2 at the same time, i.e., aspect starts contacting the spore-forming bacteria and dipicolinic acid or its salts and cationic surfactants concurrently, terminates the contact time.
(2) destination performs step 1, an embodiment of performing step 2 after step 1 completed. For example, after the step 1 to be brought into contact with the spore-forming bacteria and dipicolinic acid or a salt thereof, to remove the dipicolinic acid or a salt thereof, followed by the addition of cationic surfactant, spore-forming bacteria and a cationic surfactant it can be placed in contact and.
(3) performs the step 2 above, embodiments for performing step 1 after step 2 ends. For example, after the step 2 to be brought into contact with the spore-forming bacteria and a cationic surfactant, to remove the cationic surfactant, then adding dipicolinic acid or its salts, spore-forming bacteria and dipicolinic acid or a it can be placed in contact with the salt.
(4) destination performs step 1, without leaving step 1, mode to start the process 2. Here, it is possible to keep previously contacting a spore-forming bacteria and dipicolinic acid or a salt thereof, then dipicolinic acid or without removing a salt thereof, is contacted with a cationic surfactant.
(5) destination performs step 2, without leaving the step 2, mode to start the process 1. Here, it is possible to keep previously contacting a spore-forming bacteria and a cationic surfactant, then without removing the cationic surfactant, it is contacted with and dipicolinic acid.

Dipicolinic acid or its salts and cationic surfactants may be respectively provided in the form of liquid or solid. Further, dipicolinic acid or a salt thereof and a cationic surfactant, from the viewpoint of convenience, is preferably provided as a stop aid composition.

Here, the contact method left in contact in solution, or, if the spore-forming bacteria present on the surface solids, solution or the like containing dipicolinic acid or its salts and cationic surfactants, killing below the liquid method of applying the Mesukezai composition in solid surface thereof. As a method of coating the solid surface, a method of spraying, a method of using a tool such as a brush or sponge. Here, although not particularly limited to the solid surface, it refers to such hard surfaces.

If left in contact with the spore-forming bacteria and dipicolinic acid or a salt thereof in a solution, the content of dipicolinic acid or a salt thereof in the solution at the time of the contact, from the viewpoint of further improving the stop effect, preferably 0.05mM or more, more preferably 0.5mM or more, more preferably 3mM or more, more preferably 4mM or more, more preferably 6mM, more preferably at least 8 mM, and, the stability of solutions containing dipicolinic acid from the viewpoint, preferably not more than 1M, more preferably 200mM or less, more preferably 100mM or less, more preferably 50mM or less, more preferably 20mM or less. Moreover, if left in contact with spore-forming bacteria and dipicolinic acid or a solution in a salt thereof, content of dipicolinic acid or a salt in the solution, when taken together in view of the above, preferably, 0.05 mM ~ 1M , more preferably 0.05 ~ 200 mM, more preferably 3 ~ 100 mM, more preferably 3 ~ 25 mM. Here, if it is dipicolinic acid salts, and those obtained by converting all the content acid (hereinafter, the same meaning unless otherwise indicated).

In contact, the relationship between the initial bacteria count and dipicolinic acid or concentration of the salt is not particularly limited. For example, from the viewpoint of further improving the stop effect, dipicolinic acid or concentration of the salt to the initial bacteria number 10 8 CFU / mL is preferably 0.05mM or more, more preferably 0.5mM, more preferably at least 3mM , more preferably 4mM or more, more preferably 6mM, more preferably at least 8 mM, and, preferably not more than 1M, more preferably 200mM or less, more preferably 100mM or less, more preferably 50mM or less, more preferably it is 20mM or less.

If left in contact with the spore-forming bacteria and a cationic surfactant in solution, the content of the cationic surfactant in the solution at the time of the contact, from the viewpoint of further improving the stop effect, preferably 0. 01mM or more, more preferably 0.05mM or more, more preferably 0.1mM or more, more preferably 0.5mM or more, more preferably 3mM or more, more preferably 4mM or more, more preferably 6mM, more preferably at least 8mM , and the and preferably 1000mM or less, more preferably 500mM or less, more preferably 300mM or less, more preferably 100mM or less. Moreover, if left in contact with spore-forming bacteria and a cationic surfactant in solution, the content of the cationic surfactant in the solution, when taken together in view of the above, preferably, 0.01 ~ 1000 mM, and more preferably 0.05 ~ 1000 mM, more preferably 0.1 ~ 1000 mM, more preferably 0.5 ~ 500 mM, more preferably 3 ~ 300 mM, more preferably 4 ~ 100 mM, more preferably 6 ~ 100 mM, more preferably is 8 ~ 100mM.

If left in contact with spore-forming bacteria and a cationic surfactant in solution, the content of the cationic surfactant in the solution at the time of the contact, from the viewpoint of further improving the stop effect, preferably 0.01 mass% or more, more preferably 0.05 mass% or more, and, preferably 10 wt% or less, more preferably 5 mass% or less, more preferably not more than 1 wt%. Moreover, if left in contact with spore-forming bacteria and dipicolinic acid or a salt thereof and a cationic surfactant in solution, the content of the cationic surfactant in the solution, when taken together in view of the above, preferably 0. 01-10 wt%, more preferably from 0.05 to 5 mass%, more preferably 0.05 to 1 mass%.

In contact relationship between the concentration of the initial bacteria count and a cationic surfactant is not particularly limited. For example, from the viewpoint of further improving the stop effect, the concentration of the cationic surfactant to the initial bacteria number 10 8 CFU / mL is preferably 0.01mM or more, more preferably 0.05mM or more, more preferably 0.1mM or more, more preferably 0.5mM or more, more preferably 1mM or more, more preferably 10mM or more, more preferably 100mM or more, and, preferably 1000mM or less, more preferably 500mM or less, more preferably 300mM or less, more preferably it is 250mM or less.

These preferred concentrations, even when the advance simultaneously into contact with the spore-forming bacteria dipicolinic acid or its salts and cationic surfactants are the same even when the keep contacted separately.

In Step 3 before the start of the present invention, the temperature of step 1 or step 2 is performed independently of the step 3, from the viewpoint of further improving the stop effect, each independently preferably 0 ℃ or higher, more preferably 10 ° C. or more, more preferably 15 ℃ or more, more preferably 20 ° C. or higher, and preferably below 50 ° C., more preferably 49 ° C. or less, more preferably 45 ° C. or less, more preferably 40 ° C. or less, more preferably 30 ° C. or less. The contact temperature of the solution containing the spore-forming bacteria and dipicolinic acid or a salt or cationic surfactant in Step 1 and Step 2 is performed independently of the step 3, from the same viewpoint, preferably 15 ° C. ~ less than 50 ° C., more preferably from 15 ℃ ~ 40 ℃, more preferably 15 ~ 30 ° C..

When these preferred temperatures, even in the case of performing steps 1 and 2 at the same time, it is the same even when performed separately, in which the step 1 and step 2 are performed separately, steps 1 and second temperature may be the same or different.

Time of Step 1 or Step 2 is performed independently of the step 3 of the present invention, from the viewpoint of further improving the stop effect, each independently, preferably 1 second or more, more preferably 5 seconds or more, more preferably 30 seconds or more, more preferably 1 minute or more, more preferably 3 minutes or more, more preferably 5 minutes or more, more preferably 8 minutes or more, more preferably 10 minutes or more, more preferably 15 minutes or more, more preferably 20 minutes or more, more or preferably 25 minutes or more, and preferably 24 hours or less, more preferably 12 hours or less, more preferably 6 hours or less, more preferably 3 hours or less, more preferably 60 minutes or less, more preferably no more than 50 minutes, more preferably 40 minutes or less, and more preferably not more than 35 minutes. The contact time with the dipicolinic acid or a salt thereof and spore-forming bacteria in step 1, when taken together in view of the above, preferably 1 second to 24 hours, more preferably from 1 second to 6 hours, more preferably 1 second to 3 hours, more preferably 1 second to 60 minutes, more preferably 5 seconds to 60 minutes, more preferably from 30 seconds to 60 minutes.

<Step 3>
Step 3 of the present invention is a step of heating the spore-forming bacteria above 50 ° C..

Step 3 of the present invention is preferably a process of heating the mixture at 50 ° C. or higher 250 ° C. or less spore-forming bacteria.

Temperature for heating a spore-forming bacteria, from the viewpoint of further improving the stop effect, 50 ° C. or higher, more preferably temperatures above 55 ℃, more preferably 60 ° C. or higher, more preferably 65 ° C. or higher, more preferably 70 ° C. or more, more preferably 75 ° C. or higher, and preferably 250 ° C. or less, more preferably 200 ° C. or less, more preferably 0.99 ° C. or less, more preferably 120 ° C. or less, more preferably 100 ° C. or less, more preferably 90 ° C. or less, more preferably 85 ° C. or less. Temperature for heating in the step 3 of the present invention, in view of the above, preferably 50 ° C. ~ 250 ° C., more preferably from 50 ° C. ~ 100 ° C., more preferably from 55 ° C. ~ 100 ° C., more preferably 60 ° C. ~ 100 ° C., more preferably from 65 ℃ ~ 90 ℃, more preferably from 70 ℃ ~ 90 ℃, more preferably from 75 ℃ ~ 85 ℃.

Usually, it is difficult to sterilize in the 100 ° C. or less spore-forming bacteria are not germinate. However, in the present invention, since the germination is promoted by steps 1 and 2, stop effect even 100 ° C. or less in the step 3 is obtained. It is also possible to stop at a temperature above 100 ° C. In the present invention.

Time of heating of step 3 of the present invention, from the viewpoint of further improving the stop effect, preferably 3 minutes or more, more preferably 5 minutes or more, more preferably 8 minutes or more, more preferably 10 minutes or more, more preferably 20 minutes or more, more or preferably 25 minutes or more, and preferably 3 hours or less, more preferably less than 90 minutes, more preferably less than 70 minutes, more preferably less than 50 minutes, more preferably 40 minutes or less , more preferably not more than 35 minutes. Time of heating of step 3 of the present invention, in view of the above, preferably having 3 to 90 minutes, more preferably 5 minutes to 70 minutes, more preferably from 8 minutes to 50 minutes, more preferably 10 minutes 50 min, more preferably 20 to 40 minutes, more preferably 25 to 35 minutes. This time may comprise the steps 1 and / or step 2 and duration simultaneous.

The warming step 3 of the present invention, not particularly limited as long as it is consequently manner that the temperature of the system including the spore-forming bacteria is increased. The heating, for example, dry heat in spore-forming bacteria, moist heat, boiling, hot water, and a method of applying such steam heat. Here, for example, a method of immersing the spore-forming bacteria in hot water, a method of installing a liquid or the like in a thermostatic bath containing a solid or spore-forming bacteria such as hard surfaces in the presence of spore-forming bacteria, cooking and the like in food processing the method by heat treatment, and a method of temperature as a result, the spores in the irradiation or the like of the laser is increased and the like.

In step 1, step 2, and step 3, pH at 24 ° C. of the solution is not particularly limited, is preferably 3 or more, more preferably 4 or more, more preferably 6 or more, the safety viewpoint from, preferably 12 or less, more preferably 9 or less, more preferably 8.5 or less. Moreover, pH at 24 ° C. of the solution, when taken together in view of the above, preferably 3 to 12, more preferably 3 to 9, more preferably 4 to 8.5.

pH adjustment can be performed when previously prepared dipicolinic acid or a salt or cationic surfactant solution. Alternatively, it is also possible to carry out after addition of the dipicolinic acid or a salt or cationic surfactant in a solution containing a spore-forming bacteria when left in contact with the spore-forming bacteria. The pH adjusting agent, typically an acid or a base is used, for example, inorganic acids such as hydrochloric acid or sulfuric acid, organic acids such as lactic or citric acid or salts thereof, inorganic bases such as sodium hydroxide and potassium hydroxide, triisopropanolamine organic bases such as amines. The pH adjusting agent is preferably an inorganic acid such as hydrochloric acid or sulfuric acid as the acid, the base inorganic bases such as sodium hydroxide and potassium hydroxide are preferred.

In stop method of the present invention, at step 3 heated spore-forming bacteria, it is possible to terminate the process 3 by quenching with water was placed in ice or cooled container or the like. Alternatively, it warmed spore-forming bacteria in step 3, it is possible to terminate the process 3 by cool heat naturally after a while. Here, it warmed spore-forming bacteria may be a state that is present in the liquid.

If spore-forming bacteria are present in the liquid, it stops the process of the present invention, step 1, step 2, and before and after the step 3, or those steps may include the step of agitating the spore-forming bacteria. Step 1, Step 2, and Step 3 may be repeated.

Step 3 can be carried out from the end of the steps 1 and 2 (embodiment 1).

In embodiments 1, and "a step 3 after the completion of steps 1 and 2", is to carry out the step 3 after removing the dipicolinic acid or a salt thereof and a cationic surfactant, the step 1 and step 2 at the end, it is meant that. Embodiment 1, finishing steps 1 and 2, preferably within 24 hours, more preferably within 12 hours, more preferably within 6 hours, more preferably within 1 hour, more preferably within 30 minutes, more preferably within 15 minutes, more preferably within 5 minutes, more preferably it proceeds to step 3 within one minute.

Have a period for step 1 or step 2 and step 3 at the same time good (embodiment 2).

In Embodiment 2, a "has a period for step 1 or step 2 and step 3 at the same time", is to carry out the step 3 after removing the dipicolinic acid or a salt or cationic surfactant, Step 1 or Step 2 together with the end, it is meant that. Embodiment 2, finishing step 1 or step 2, preferably within 24 hours, more preferably within 12 hours, more preferably within 6 hours, more preferably within 1 hour, more preferably within 30 minutes, more preferably within 15 minutes, more preferably within 5 minutes, more preferably it proceeds to step 3 within one minute.

Step 1 and Step 2 and Step 3 may have a period performed simultaneously (mode 3). Mode 3 is divided as follows.
Embodiments 3-1 Step 1 and / or step 2, above aspects 3-2 steps 1 and 2 and the embodiment 3-3 Step 3 to begin the process 3 simultaneously to start before the step 3 from step 1 and step 2 among the embodiments 3-4 step 1 or step 2 and step 3 aspects 3 to start at the same time beginning, from the viewpoint of further improving the stop effect, it is preferred that embodiments 3-1 but is not limited thereto.

Stop method of the present invention, from the viewpoint of further improving the stop method, an embodiment having a period in which step 1 and step 2 and step 3 at the same time (aspect 3) is preferable.

<Yamesuke composition>
Killing bud aid composition of the present invention is dipicolinic acid or compositions comprising the salts and cationic surfactants. Killing bud aid composition of the present invention performs the purpose of germination and used warming pretreatment. Moreover, it is distinguished from leave composition that aims to stop using the composition itself at room temperature (about 20 ° C. ~ about 38 ° C.). In particular, after the step 1 and step 2 of the present invention, or simultaneously with step 1 and step 2, it refers to a composition capable of exhibiting stop effect by warming. In the present invention, Yamesuke composition may be used in step 1 and step 2.

The Yamesuke compositions, liquid compositions or solid compositions.

Stop aid composition of liquid comprises dipicolinic acid or a salt thereof, a cationic surfactant, and a solvent.

If Yamesuke composition is a liquid, as the solvent contained include water or a hydrophilic solvent. The hydrophilic solvents ethanol, methanol, monohydric alcohols such as isopropanol; glycerin, ethylene glycol, polyhydric alcohols such as propylene glycol; methyl carbitol, and carbitol and ethyl carbitol and the like. Solvent, from the viewpoint of further improving the stop effect, preferably water or mixtures of water and a hydrophilic solvent, more preferably water.

The stop aid composition of the liquid, from the viewpoint of convenience, preferably a solution containing dipicolinic acid or a salt thereof and a cationic surfactant, more preferably an aqueous solution containing a dipicolinic acid or a salt thereof and a cationic surfactant is there.

Stop aid composition of the liquid can further contain a pH adjusting agent. The pH adjusting agent contained in the stop aid composition of the liquid, generally acids used and bases, for example, inorganic acids such as hydrochloric acid or sulfuric acid, lactic acid and citric acid or an organic acid such as their salts, sodium hydroxide and inorganic bases such as potassium hydroxide, monoethanolamine, triethanolamine, and organic bases such as triisopropanolamine. The pH adjusting agent is preferably an inorganic acid such as hydrochloric acid or sulfuric acid as the acid, the base inorganic bases such as sodium hydroxide and potassium hydroxide are preferred.

Stop aid composition of the liquid, preferably pH at 24 ° C. of an aqueous solution containing a solution comprising dipicolinic acid or its salts and cationic surfactants, more preferably dipicolinic acid or its salts and cationic surfactants are particularly limited but are not, is preferably 3 or more, more preferably 4 or more, more preferably 6 or more, from the viewpoint of safety, is preferably 12 or less, more preferably 9 or less, more preferably 8.5 less. Moreover, pH at 24 ° C. of the solution, when taken together in view of the above, preferably 3 to 12, more preferably 3 to 9, more preferably 4 to 8.5.

Stop aid composition of the liquid content of preferably dipicolinic acid in aqueous solution containing in solution comprising dipicolinic acid or its salts and cationic surfactants, more preferably dipicolinic acid or a salt thereof and a cationic surfactant from the viewpoint of further improving the stop effect, preferably 0.05mM or more, more preferably 0.5mM or more, more preferably 3mM or more, more preferably 4mM or more, more preferably 6mM, more preferably at least 8mM and a, and, from the viewpoint of stability of the solution containing the dipicolinic acid or its salts and cationic surfactants, preferably not more than 1M, more preferably 200mM or less, more preferably 100mM or less, more preferably 50mM or less, more preferably 20mM or less. The content of dipicolinic acid or a salt stop aid composition in liquid Taken together the above viewpoint, preferably, 0.05 mM ~ 1M, more preferably 0.05 mM ~ 200 mM, more preferably , 3 mM ~ 100 mM, more preferably 3 mM ~ 15 mM, more preferably 4 mM ~ 15 mM.

Stop aid composition of the liquid is preferably in a solution containing dipicolinic acid or a salt thereof and a cationic surfactant, more preferably a cationic surfactant in an aqueous solution containing dipicolinic acid or its salts and cationic surfactants content, from the viewpoint of further improving the stop effect, preferably 0.05mM or more, more preferably 0.5mM or more, more preferably 3mM or more, more preferably 4mM or more, more preferably 6mM or more, more preferably and at 8mM more, and preferably not more than 1M, more preferably 200mM or less, more preferably 100mM or less, more preferably 50mM or less, more preferably 20mM or less. The content of cationic surfactants stop aid composition in liquid Taken together the above viewpoint, preferably 0.1 ~ 1M, more preferably 0.5 ~ 500 mM, more preferably 3 to 300 mM, more preferably 4 ~ 100 mM, more preferably 6 ~ 100mM, 8 ~ 100mM.

The content of cationic surfactants stop aid composition in the liquid, from the viewpoint of further improving the stop effect, preferably 0.01 mass% or more, more preferably 0.05 mass% or more, and, preferably 10 wt% or less, more preferably 5 mass% or less, more preferably not more than 1 wt%. The content of cationic surfactants stop aid composition of the liquid from the overall aspects of the above, preferably 0.01 to 10 mass%, more preferably from 0.05 to 5 mass%, more preferably is 0.05 to 1 mass%.

Killing bud aid composition of the present invention, dipicolinic acid or its salts and cationic surfactants, including water or other solvents, peracetic acid, sulfo peroxycarboxylic acid or a composition containing neither dicarboxylic acid diester, possible. Here it does not include a indicates that Yamesuke composition has no stop effect, preferably 1000ppm or less, more preferably 100ppm or less, more preferably 10ppm or less, more preferably 1ppm or less, more preferably it is substantially 0ppm. Here it is below the detection limit substantially.

Stop aid composition of the liquid of the present invention, preferably a solution containing dipicolinic acid or its salts and cationic surfactants, aqueous solutions more preferably comprising dipicolinic acid or its salts and cationic surfactants, the powdery dipicolinic an acid or a salt or cationic surfactant by mixing with water or other solvents, can preferably be prepared by dissolving.

The stop aid composition of solid, dipicolinic acid or a salt thereof and a cationic surfactant or dipicolinic acid or a salt thereof, a cationic surfactant, and compositions comprising solidifying agents.

The solidifying agent to stop aid composition solids are contained, but are not limited to, polyethylene glycol having a number average molecular weight of 1,000 to 100,000; carnauba wax, candelilla wax, jojoba oil , beeswax, waxes such as lanolin; paraffin, vaseline, ceresin, hydrocarbons having 15 or more carbon atoms, such as microcrystalline wax; lauric acid, myristic acid, a higher fatty acid having 12 to 22 carbon atoms such as stearic acid; cetyl alcohol, higher alcohols having 14 to 22 carbon atoms, such as stearyl alcohol.

Stop aid composition of the solid can further contain a pH adjusting agent. The pH adjusting agent stop aid composition of solids contains, generally acids used and bases, for example, inorganic acids such as hydrochloric acid or sulfuric acid, lactic acid and citric acid or an organic acid such as their salts, sodium hydroxide and inorganic bases such as potassium hydroxide, monoethanolamine, triethanolamine, and organic bases such as triisopropanolamine. The pH adjusting agent is preferably an inorganic acid such as hydrochloric acid or sulfuric acid as the acid, the base inorganic bases such as sodium hydroxide and potassium hydroxide are preferred.

Killing bud aid composition of the present invention, when used in step 1 and step 2 of stop method of the present invention, the content of dipicolinic acid solution containing spore-forming bacteria and stop aid composition from the viewpoint of further improving the stop effect, preferably 0.05mM or more, more preferably 0.5mM or more, more preferably 3mM or more, more preferably 4mM or more, more preferably 6mM or more, more preferably at least 8mM There, and in view of the stability of solutions containing dipicolinic acid and cationic surfactants, preferably 200mM or less, more preferably 100mM or less, more preferably 30mM or less, more preferably 25mM or less, more preferably 20mM or less it is preferred to use a certain manner. Moreover, killing buds aid composition of the present invention, when used in step 1 and step 2 of stop method of the present invention, the content of dipicolinic acid solution containing spore-forming bacteria and stop aid composition amount, when taken together in view of the above, preferably 0.05 ~ 200 mM, more preferably 0.5 ~ 100 mM, more preferably 3 ~ 30 mM, it is preferred to use as more preferably at 3 ~ 25 mM.

Stop aid composition of the liquid, when used in step 1 and step 2 of stop method of the present invention, the content of cationic surfactant solution containing spore-forming bacteria and stop aid composition but preferably a solution containing dipicolinic acid and cationic surfactants, and more preferably the content of the cationic surfactant in the aqueous solution containing dipicolinic acid and cationic surfactants, from the viewpoint of further improving the stop effect, preferably 0.1mM or more, more preferably 0.5mM or more, more preferably 3mM or more, more preferably 4mM or more, more preferably 6mM, more preferably at least 8 mM, and, preferably 1000mM or less, more preferably is 500mM or less, more preferably 300mM or less, more preferably 100mM or less. The content of cationic surfactants in the use of stop aid composition of the liquid from the overall aspects of the above, preferably 0.1 ~ 1000 mM, more preferably 0.5 ~ 500 mM, and more preferably the 3 ~ 300 mM, more preferably 4 ~ 100 mM, more preferably 6 ~ 100 mM.

Stop aid composition of the liquid, when used in step 1 and step 2 of stop method of the present invention, the content of cationic surfactant solution containing spore-forming bacteria and stop aid composition but from the viewpoint of further improving the stop effect, preferably 0.01 mass% or more, more preferably 0.05 mass% or more, and, preferably 10 wt% or less, more preferably 5 mass% or less, more preferably is less than 1 wt%. The content of cationic surfactants in the use of stop aid composition of the liquid from the overall aspects of the above, preferably 0.01 to 10 mass%, more preferably from 0.05 to 5 mass %, more preferably 0.05 to 1 mass%.

Dipicolinic acid or a salt thereof of the present invention, by combining, or although by fermentation method to use a method of extracting from microorganisms, either can be used. Cationic surfactants of the present invention include those which are commercially available, none of the by synthesis may be used.

Stop methods and stop aid composition of the present invention, sterilizing and washing and food processing equipment, can be used for sterilization such as washing of the garment.

Relates the above-described embodiment, the present invention discloses the following stop method and stop aid composition.

<Claim 1>
The following steps 1, quit method of performing steps 2 and 3:
Step 1: a step of previously contacting the spore-forming bacteria and dipicolinic acid or a salt thereof;
Step 2: Step left in contact with the spore-forming bacteria and a cationic surfactant; and Step 3: a step of heating the spore-forming bacteria above 50 ° C. (provided that terminates the process 3 after starting the steps 1 and 2 It shall be).

<Claim 2>
Step 1, Step 2, and a period for step 3 simultaneously, <claim 1> quit method according.

<Section 3>
Step perform steps 2 and 3 after one end, <claim 1> quit method according.

<Claim 4>
Step perform steps 1 and 3 after 2 termination, <claim 1> quit method according.

<Section 5>
Cationic surfactant, a primary ammonium salt, a secondary ammonium salt is either a tertiary ammonium salt or a quaternary ammonium salt, according to any one of <claim 1> to <claim 4> quit method.

<Section 6>
Cationic surfactant is an alkyltrimethylammonium salt, dialkyldimethylammonium salt, benzalkonium salt, alkyl pyridinium salt, quaternary ammonium salts such as alkyl benzethonium salts, primary ammonium salts such as alkylamine salts, < claim 1> to quit method according to any one of <5.>.

<Section 7>
Cationic surfactant is a quaternary ammonium salt represented by the general formula (1), <claim 1> to quit method according to any one of <6.>.

Figure JPOXMLDOC01-appb-C000005

Wherein any one of R 11 ~ R 14, hydroxyl group, an ester group, an amide group hydrocarbon group having 3 or more carbon atoms which may have a remaining three of R 11 ~ R 14 represents a methyl group or an ethyl group, X - represents an inorganic or organic anionic compounds. ]

<Section 8>
Cationic surfactants, alkyltrimethylammonium salts, alkyl triethyl ammonium salts, alkyl dimethyl ethyl ammonium salt, alkyl methyl diethyl ammonium salt, dialkyl dimethyl ammonium salts, dialkyl diethyl ammonium salts, dialkyl methyl ammonium salts, benzalkonium salts, alkyl it is at least one selected from the pyridinium salt and the group consisting of alkyl benzethonium salt, <claim 1> to quit method according to any one of <claim 7>.

<Section 9>
In the step 1 and step 2, spore-forming bacteria, placed in contact dipicolinic acid or a salt thereof, and a cationic surfactant in a liquid, <claim 1> to quit method according to any one of <section 8>.

<10.>
Stop method of the content of dipicolinic acid or its salt is 200mM or less than 0.05mM in the liquid <claim 9> wherein.

<11.>
The content of the cationic surfactant in the solution is less than 1000mM than 0.1 mM, <9.> quit method according.

<12.>
Spore-forming bacteria and dipicolinic acid or its dipicolinic acid or concentration of a salt thereof when salt is in contact above 0.05 mM, and more preferably 0.5mM or more, more preferably 3mM or more, more preferably 4mM more, more preferably 6mM or more, more preferably at least 8 mM, and, preferably 200mM or less, more preferably 100mM or less, more preferably 50mM or less, more preferably 20mM or less, wherein one of the <claim 1> to <claim 11> quit method of.

<Section 13>
The concentration of the cationic surfactant upon contact between the spore-forming bacteria and a cationic surfactant, 0.1 mM or higher, more preferably 0.5mM or more, more preferably 3mM or more, more preferably 4mM or more, more preferably 6mM, more preferably at least 8 mM, and, preferably 1000mM or less, more preferably 500mM or less, more preferably 300mM or less, more preferably 100mM or less, either <claim 1> to <claim 12> quit the method described.

<14.>
The concentration of the cationic surfactant upon contact between the spore-forming bacteria and a cationic surfactant, 0.01% by mass or more, more preferably 0.05 mass% or more, and, preferably 10 wt% or less, more preferably 5 wt% or less, much more preferably 1 wt%, <claim 1> to quit method according to any one of <claim 13> is.

<15.>
The steps 1 and 2, carried out in each less than independently or simultaneously 15 ℃ above 50 ° C., <claim 1> to quit method according to any one of <claim 14>.

<16.>
The Steps 1 and 2 of the step 3 is performed independently or simultaneously each independently, 15 ° C. or more, more preferably 20 ° C. or higher, and preferably below 50 ° C., more preferably 49 ° C. or less , more preferably 45 ° C. or less, more preferably 40 ° C. or less, and more preferably carried out at 30 ° C. or less, <claim 1> to quit method according to any one of <claim 15>.

<Section 17>
Step 3 time steps 1 performed independently of, i.e., contact time with the spore-forming bacteria and dipicolinic acid or its salts, preferably one second or more, more preferably 5 seconds or more, more preferably 30 seconds or more , more preferably at least 1 minute, more preferably 3 minutes or more, more preferably 5 minutes or more, more preferably 8 minutes or more, more preferably 10 minutes or more, more preferably 15 minutes or more, more preferably 20 minutes or more, more is preferably 25 minutes or more, and preferably 60 minutes or less, more preferably less than 50 minutes, more preferably 40 minutes or less, and more preferably not more than 35 minutes, the <claim 1> to <claim 16> quit method according to any one.

<18.>
Time Step 2 when performed independently of the step 3, i.e., the contact time with the spore-forming bacteria and a cationic surfactant, preferably at least 1 second, more preferably 5 seconds or more, more preferably 30 seconds or more , more preferably at least 1 minute, more preferably 3 minutes or more, more preferably 5 minutes or more, more preferably 8 minutes or more, more preferably 10 minutes or more, more preferably 15 minutes or more, more preferably 20 minutes or more, more is preferably 25 minutes or more, and preferably 60 minutes or less, more preferably less than 50 minutes, more preferably 40 minutes or less, and more preferably not more than 35 minutes, the <claim 1> to <claim 17> quit method according to any one.

<19.>
The total contact time for contacting the spore-forming bacteria and dipicolinic acid or a salt thereof in step 1 when having a period concurrently with step 3 is preferably 5 minutes or more, more preferably 10 minutes or more, more preferably 15 minutes or more, more preferably 20 minutes or more, more or preferably 25 minutes or more, and preferably less than 90 minutes, more preferably at most 80 minutes, more preferably less than 70 minutes, more preferably 65 minutes or less, more preferably 60 minutes or less, more preferably less than 50 minutes, more preferably 40 minutes or less, and more preferably not more than 35 minutes, <claim 1> to quit method according to any one of <claim 18>.

<20.>
The total contact time of spore-forming bacteria and a cationic surfactant in step 2 in the case with a period in which at the same time as step 3 is preferably 5 minutes or more, more preferably 10 minutes or more, more preferably 15 minutes or more, more preferably 20 minutes or more, more is preferably 25 minutes or more, and preferably 90 minutes or less, more preferably less than 80 minutes, more preferably less than 70 minutes, more preferably less 65 minutes, more preferably 60 minutes or less, more preferably not more than 50 minutes, more preferably 40 minutes or less, and more preferably not more than 35 minutes, <claim 1> to quit method according to any one of <claim 19>.

<21.>
Time for heating the spore-forming bacteria above 50 ° C. In Step 3, is 90 minutes or less than 3 minutes, <claim 1> to <claim 20> stop method of any.

<22.>
Temperature of step 3 is preferably 50 ° C. or higher, more preferably temperatures above 55 ℃, more preferably 60 ° C. or higher, more preferably 65 ° C. or higher, more preferably 70 ° C. or more, more preferably 75 ° C. or higher, and, preferably 250 ° C. or less, more preferably 200 ° C. or less, more preferably 0.99 ° C. or less, more preferably 120 ° C. or less, more preferably 100 ° C. or less, more preferably 90 ° C. or less, more preferably 85 ° C. or less, <claim 1> to quit method according to any one of <claim 21>.

<23.>
Time of heating of step 3 is preferably 3 minutes or more, more preferably 5 minutes or more, more preferably 8 minutes or more, more preferably 10 minutes or more, more preferably 20 minutes or more, more preferably at least 25 minutes There, and preferably less than 90 minutes, more preferably less than 70 minutes, more preferably less than 50 minutes, more preferably 40 minutes or less, and more preferably not more than 35 minutes, the <claim 1> to <claim 22> quit method according to any one.

<24.>
Wherein the step 3 before the start, with a period for performing the step 1 and step 2 respectively below independently or simultaneously 15 ℃ above 50 ° C., <claim 1> to quit method according to any one of <claim 23>.

<25.>
Step 3 before the start, the process performed 1 and step 2 respectively independently or simultaneously 60 minutes or less than 1 sec, <claim 1> to quit method according to any one of <claim 24>.

<26.>
The process performed 1 and step 2 are each independently a total or simultaneously more than 30 minutes 90 minutes or less, <claim 1> to quit method according to any one of <claim 25>.

<27.>
Dipicolinic acid or a salt thereof and a cationic surfactant, Yamesuke composition.

<28.>
Dipicolinic acid or its salt preferably 0.05mM or more, more preferably 0.5mM or more, more preferably 3mM more, and, preferably 200mM or less, more preferably 100mM or less, more preferably a liquid composition containing 30mM or less in it, <claim 27> stop aid composition.

<29.>
Cationic surfactant preferably 0.1mM or more, more preferably 0.5mM or more, more preferably 3mM more, and, preferably 1000mM or less, more preferably 500mM or less, more preferably with a liquid composition containing 300mM or less there, <claim 27> or <claim 28> stop aid composition.

<30.>
Dipicolinic acid or a salt thereof, a cationic surfactant, and includes water or other solvents, the content of peracetic acid, sulfo peroxycarboxylic acid and dicarboxylic acid diester is preferably 1000ppm or less, more preferably 100ppm or less, more preferably 10ppm or less, more preferably 1ppm or less, and more preferably substantially 0 ppm, <claim 27> ~ quit aid composition according to any one of <claim 29>.

Hereinafter, Examples illustrate the present invention more specifically.

Preparation spore-forming bacteria of bacteria were used: Bacillus subtilis 168 strain as a test strain. We used in the experiment the content of spore-forming bacteria to prepare a water dispersion having a 10 8 CFU / mL. Spore-forming bacteria of the aqueous dispersion, by microscopic observation, 95% or more is used in the experiments after confirming that forming spores.

Dipicolinic acid solution prepared dipicolinic acid (hereinafter also referred to as DPA) reagent (manufactured by Wako Pure Chemical Industries, Ltd., manufacturing code 165-05342) and, Tris-HClbuffer (Wako Junyaku solvent adjusted with deionized water to 50mM industry Co., Ltd., a manufacturing code 318-90225), pH adjusting agent with sodium hydroxide. Preparation of dipicolinic acid aqueous solution was carried out at 24 ° C.. While measuring the pH of dipicolinic acid aqueous solution at benchtop pH meter Model 9611 (manufactured by HORIBA), it was added dropwise a pH adjusting agent dipicolinic acid aqueous solution to adjust the pH of the dipicolinic acid solution. In the examples, unless otherwise indicated, pH is a pH at 24 ° C.. Unless otherwise specified, adjusted to pH 7 (24 ° C.), an experiment was conducted.

Using a surfactant of a surfactant.
Lauryl trimethyl ammonium chloride (Quartamin 24P, manufactured by Kao Corporation)
Alkyl benzyl dimethyl ammonium chloride (Sanizo - manufactured by Le C Kao Corporation)
Sodium lauryl sulfate (Emal 0, manufactured by Kao Corporation)
Polyoxyethylene (3) lauryl ether sulfate (EMAL 20C, manufactured by Kao Corporation)
Polyoxyethylene (2) Raurirue one ether sodium sulfate (EMAL E-27C, manufactured by Kao Corporation)
Sodium dodecyl benzene sulfonate (Neo Beck G-15, manufactured by Kao Corp.)
Polyoxyethylene lauryl ether (Emulgen 108, manufactured by Kao Corporation)
Polyoxyethylene (23) lauryl ether (Emulgen 123P, manufactured by Kao Corporation)
Polyoxyethylene lauryl ether (Emulgen 130K, manufactured by Kao Corporation)
Polyethylene glycol monolaurate (Emanon 1112, Kao Corporation)
Polyoxyethylene hydrogenated castor oil (60E.O.) (Emanon CH-60, manufactured by Kao Corp.)
Lauryl glucoside (My Doll 12, manufactured by Kao Corp.)
Dodecyltrimethylammonium chloride (manufactured by Tokyo Kasei, C12 trimethyl ammonium chloride below)
Tetradecyl trimethyl ammonium chloride (manufactured by Tokyo Kasei, hereinafter C14 trimethyl ammonium chloride)
Hexadecyltrimethylammonium chloride (manufactured by Tokyo Kasei, hereinafter C16 trimethyl ammonium chloride)
Octadecyl trimethyl ammonium chloride (manufactured by Tokyo Kasei, hereinafter C18 trimethyl ammonium chloride)
Dioctanylphosphatidyl dimethyl ammonium chloride (manufactured by Tokyo Kasei, hereinafter C8 dimethyl ammonium chloride)
Didecyl dimethyl ammonium chloride (manufactured by Tokyo Kasei, hereinafter C10 dimethyl ammonium chloride)
Didodecyl dimethyl ammonium chloride (manufactured by Tokyo Kasei, hereinafter C12 dimethyl ammonium chloride)
Ditetradecyldimethylammonium chloride (manufactured by Tokyo Kasei, hereinafter C14 dimethyl ammonium chloride)
Dihexadecyl dimethyl ammonium chloride (manufactured by Tokyo Kasei, hereinafter C16 dimethyl ammonium chloride)
Dioctadecyl dimethyl ammonium chloride (manufactured by Tokyo Kasei, hereinafter C18 dimethyl ammonium chloride)

Example 1-1:
Stop 10mM dipicolinic acid solution as described in the test Table 1 (hereinafter, also referred to as DPA solution), using a 0.10 wt% lauryl trimethyl ammonium chloride and spore-forming bacteria, subjected to quit tested by the following procedure, It was evaluated stop effect. The evaluation results are shown in Table 1. (In the table, the term "%" is "% by mass". The same applies hereinafter.)

-procedure-

(1) Step 1 and Step 2: aqueous dispersion of spore-forming bacteria; mixing 100 [mu] L (hereinafter, referred to as spore solution same in the following examples), and the DPA solution 800 [mu] L, an aqueous solution 100 [mu] L of lauryl trimethyl ammonium chloride and to obtain a test solution. The final concentration of DPA is 10 mM, the final concentration of lauryl trimethyl ammonium chloride was 0.1 wt%. Here, the final concentration refers to the concentration in the aqueous solution after mixing (state a total of 1 mL) (same in the following examples).
(2) Step 1 and Step 2: the test solution described above (1) to (pH 7.0) was allowed to stand for 30 minutes at 24 ° C..
(3) Step 3: After the above (2), the test solution was 30 minutes warming treated as 80 ° C..
The heating step was used Major Science's aluminum block thermostatic chamber MD-01N-110.
(4) LP solution (LP diluent (Daigo, Nippon Pharmaceutical Co., Ltd.)) was heat-treated test solution in (3) were serially diluted in, 100 [mu] L smeared each dilution on LB agar medium (manufactured by BD) and, 37 ℃, were cultured for 16 hours.
(5) (4) After the culture, at the number has grown colonies was determined viable cell count X (CFU / mL).

Comparative Example 1-1 and 1-2
In Step (1) of Example 1-1, instead of an aqueous solution containing a final concentration of 10 mM DPA and final concentration of 0.1 wt% cationic surfactant, using an aqueous solution shown in Table 1. Otherwise it was conducted in the same manner as in Example 1-1.

Figure JPOXMLDOC01-appb-T000006

Examples 2-1 and Comparative Examples 2-1 to 2-10:
The DPA solution, using an aqueous solution obtained by mixing the aqueous surfactant solution shown in Table 2. The final concentration of DPA in the aqueous solution is 10 mM, the final concentration of each surfactant was 0.1 wt%. Under the same conditions as in Example 1-1, performs stop tests were evaluated stop effect. The evaluation results are shown in Table 2.

Figure JPOXMLDOC01-appb-T000007

Examples 3-1 to 3-10:
The DPA solution, using an aqueous solution obtained by mixing the aqueous surfactant solution shown in Table 3. DPA and the final concentration of each surfactant in the aqueous solution was 10 mM. Performs stop test under the same conditions as in Example 1-1, it was evaluated stop effect. The evaluation results are shown in Table 3.

Comparative Examples 3-1 to 3-3:
Except that the temperature in step 3 was RT (25 ° C.) was evaluated stop effect under the same conditions as in Example 3-1,3-3,3-4.

Figure JPOXMLDOC01-appb-T000008

Examples 4-1 to 4-13:
The DPA solution, using an aqueous solution obtained by mixing the aqueous surfactant solution shown in Table 4. The final concentration of DPA in the aqueous solution is 10 mM, the final concentration of each surfactant was concentrations shown in Table 4. Performs stop test under the same conditions as in Example 1-1, it was evaluated stop effect. The evaluation results are shown in Table 4.

Figure JPOXMLDOC01-appb-T000009

Examples 5-1 to 5-3:
The DPA solution, using an aqueous solution of a mixture of C18 trimethyl ammonium chloride aqueous solution. The final concentration of DPA in the aqueous solution is 10 mM, C18 final concentration of trimethylammonium chloride was 20 mM. Procedure warming temperature in (3) was the temperature shown in Table 5. Performs stop test under the same conditions as in Example 1-1, it was evaluated stop effect. The evaluation results are shown in Table 5.

Figure JPOXMLDOC01-appb-T000010

Examples 6-1 to 6-4
The DPA solution, using an aqueous solution of a mixture of C18 trimethyl ammonium chloride aqueous solution. The final concentration of DPA in the aqueous solution is 10 mM, C18 final concentration of trimethylammonium chloride was 1 mM. Procedure (1) and the pH of the test solution in (2) (24 ° C.) was a value shown in Table 6. Performs stop test under the same conditions as in Example 1-1, it was evaluated stop effect. The evaluation results are shown in Table 6. Example 6-2 buffer system sodium hydroxide and phthalic acid, a buffer system of Example 6-3 boric acid sodium hydroxide, examples 6-4 and potassium chloride buffer system of sodium hydroxide Using.

Figure JPOXMLDOC01-appb-T000011

Example 7-1
-procedure-
(1) Step 1: The spore solution 100 [mu] L, were mixed with DPA solution 900 [mu] L, were tested liquid with (pH 7.0). The final concentration of the DPA was 10mM.
(2) The test was allowed to stand for 30 minutes at 24 ° C..
(3) removing the DPA solution from spores by the following method. Of test solution step 1 centrifuge subjected to centrifugal separation under the conditions of (Kubota Corporation Ltd. tabletop micro refrigerated centrifuge 3500, hereinafter the same) in 15krpm5 minutes, the supernatant was removed.
(4) Step 2: Remove the supernatant immediately a C16 trimethyl ammonium chloride aqueous solution was added 1mL to a final concentration of 10 mM, and allowed to stand for 30 minutes at 24 ° C..
(5) Step 3: After the above (4), the test solution was heated for 30 minutes at 80 ° C. The.
(6) (5) of the test solution was serially diluted with sterile water, each dilution was 100μL spread on an LB agar medium (manufactured by BD Co.), 37 ° C., and cultured for 16 hours.
(7) after (6) culture, in the number has grown colonies was determined viable cell count X (CFU / mL).

Example 7-2
-procedure-
(1) Step 2: the spore solution 100 [mu] L, were mixed with C16 trimethyl ammonium chloride aqueous solution 900 [mu] L, were tested liquid with (pH 7.0). C16 final concentration of trimethylammonium chloride was 10 mM.
(2) The test was allowed to stand for 30 minutes at 24 ° C..
(3) by the following method to remove C16 trimethyl ammonium chloride solution from spores. The test liquid (2) subjected to centrifugal separation under the conditions of 15krpm5 minutes centrifuge, the supernatant was removed.
(4) Step 1: Remove the supernatant immediately, 1 mL added DPA solution to a final concentration of 10 mM, and allowed to stand for 30 minutes at 24 ° C..
(5) After the above (4), the test solution was heated for 30 minutes at 80 ° C. The.
(6) (5) of the test solution was serially diluted with sterile water, each dilution was 100μL spread on an LB agar medium (manufactured by BD Co.), 37 ° C., and cultured for 16 hours.
(7) after (6) culture, in the number has grown colonies was determined viable cell count X (CFU / mL).

Example 7-3
-procedure-
(1) Step 1 and Step 2: the spore solution 100 [mu] L, mixing the DPA and C16 trimethyl ammonium chloride aqueous chloride 900 [mu] L, were tested liquid with (pH 7.0). DPA and C16 final concentration of trimethylammonium chloride was 10 mM.
After (2) (1), immediately test solution was 30 minutes heating treatment at 80 ° C. The.
(3) (2) The test solution was serially diluted with sterile water, each dilution was 100μL spread on an LB agar medium (manufactured by BD Co.), 37 ° C., and cultured for 16 hours.
(4) (3) After the culture, at the number has grown colonies was determined viable cell count X (CFU / mL).

Comparative Example 7-1
-procedure-
(1) The spore solution 100 [mu] L was mixed with DPA solution 900 [mu] L, were tested liquid with (pH 7.0). The final concentration of the DPA was 10mM.
After (2) (1), immediately test solution was 30 minutes heating treatment at 80 ° C. The.
(3) by the following method to remove DPA solution from spores.
The test liquid (2) subjected to centrifugal separation under the conditions of 15krpm5 minutes centrifuge, the supernatant was removed.
(4) Remove the supernatant immediately, 1 mL added C16 trimethyl ammonium chloride aqueous solution to a final concentration of 10 mM, and allowed to stand for 30 minutes at 24 ° C..
(5) (4) the test liquid was serially diluted with sterile water, each dilution was 100μL spread on an LB agar medium (manufactured by BD Co.), 37 ° C., and cultured for 16 hours.
(6) (5) After the culture, at the number has grown colonies was determined viable cell count X (CFU / mL).

Comparative Example 7-2
-procedure-
(1) Step 3: the spore solution 100 [mu] L, was heated for 30 minutes at 80 ° C..
(2) Step 2: After (1), after cooling to room temperature, immediately mixed with C16 trimethyl ammonium chloride aqueous solution 900 [mu] L, allowed to stand for 30 minutes, and test liquid and (pH 7.0). C16 final concentration of trimethylammonium chloride was 10 mM.
(3) (2) The test solution was serially diluted with sterile water, each dilution was 100μL spread on an LB agar medium (manufactured by BD Co.), 37 ° C., and cultured for 16 hours.
(4) (3) After the culture, at the number has grown colonies was determined viable cell count X (CFU / mL).

Figure JPOXMLDOC01-appb-T000012

Example 8-1:
In the same manner as in Example 3-1 was carried out stop test.

Comparative Example 8-1:
Instead of DPA solution was used isophthalic acid aqueous solution of the same concentration. Otherwise it was conducted in the same manner as in Example 8-1.

Figure JPOXMLDOC01-appb-T000013

Example 9-1
(1) Step 1 and Step 2: the spore solution 100 [mu] L, and the DPA solution 800 [mu] L, and an aqueous solution 100μL of C16 trimethyl ammonium chloride, each mixed, test solution and (pH 7.0). DPA and C16 final concentration of trimethylammonium chloride was 10 mM.
(2) Step 3: After the above (1), and immediately the test solution for 30 minutes warming treated as 80 ° C. to. The heating step was used Major Science's aluminum block thermostatic chamber MD-01N-110.
(3) above (2) LP aqueous heat-treated test solution in (LP diluent (Daigo, Nippon Pharmaceutical Co., Ltd.)) were serially diluted, 100 [mu] L of each dilution on LB agar medium (manufactured by BD) smeared, 37 ℃, were cultured for 16 hours.
(4) (3) After the culture, at the number has grown colonies was determined viable cell count X (CFU / mL).

Comparative Example 9-1
In similar operation as in Example 9-1, but without heating of step 3 was allowed to stand for 30 minutes at room temperature.

Comparative Example 9-2
Instead of (1) DPA and cationic surfactant solution of Step 1 and Step 2 of Example 9-1 was contacted spores liquid and purified water. Heating of step 3 was carried out in the same manner as in Example 9-1.

Example 9-1 are shown in Table 9 the results of Comparative Examples 9-1 to 9-2.

Figure JPOXMLDOC01-appb-T000014

Examples 10-1 to 10-2
The DPA solution, using an aqueous solution of a mixture of C16 trimethyl ammonium chloride aqueous solution. The final concentration of DPA is the concentration shown in Table 10, the final concentration of C16 trimethyl ammonium chloride was 10 mM. Performs stop test under the same conditions as in Example 1-1, it was evaluated stop effect. The evaluation results are shown in Table 10.

Figure JPOXMLDOC01-appb-T000015

Stop method of the present invention, for example, it can be used linens cleaning, preventing food spoilage, in a wide range of applications, such as environmental clean.

Claims (12)

  1. Following steps 1, step 2, and stop comprising the step 3:
    Step 1: a step of previously contacting the spore-forming bacteria and dipicolinic acid or a salt thereof;
    Step 2: Step left in contact with the spore-forming bacteria and a cationic surfactant; and Step 3: a step of heating the spore-forming bacteria above 50 ° C. (provided that terminates the process 3 after starting the steps 1 and 2 It shall be).
  2. The step 1, step 2, and step has a duration which simultaneously 3, stop the process of claim 1.
  3. The cationic surfactant is a quaternary ammonium salt, according to claim 1 or 2 stop method according.
  4. The cationic surfactant is a quaternary ammonium salt represented by the general formula (1), according to claim 3 stop method according.
    Figure JPOXMLDOC01-appb-C000001

    Wherein any one of R 11 ~ R 14, hydroxyl group, an ester group, an amide group hydrocarbon group having 3 or more carbon atoms which may have a remaining three of R 11 ~ R 14 represents a methyl group or an ethyl group, X - represents an inorganic or organic anionic compounds. ]
  5. The quaternary ammonium salt, alkyl trimethyl ammonium salt is at least one selected from the group consisting of dialkyl dimethyl ammonium salts and benzalkonium salts, according to claim 3 stop method according.
  6. In the step 1 and step 2, spore-forming bacteria, placed in contact dipicolinic acid or a salt thereof, and a cationic surfactant in a liquid, stop process of any one of claims 1 to 5.
  7. The content of dipicolinic acid or a salt thereof in the solution is 200mM or less than 0.05 mM, quit method of claim 6 wherein.
  8. The content of the cationic surfactant in the solution is less than 1000mM than 0.1 mM, claim 6 or 7 quit method according.
  9. The time to heat the spore-forming bacteria above 50 ° C. In Step 3, is 90 minutes or less than 3 minutes, stop the method of any one of claims 1-8.
  10. Dipicolinic acid or a salt thereof and a cationic surfactant, Yamesuke composition.
  11. Dipicolinic acids or liquid composition containing 0.05mM or more 200mM less salt thereof, according to claim 10 stop aid composition.
  12. The cationic surfactant which is a liquid composition containing 0.1mM than 1000mM less, claim 10 or 11 stop aid composition.
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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030175318A1 (en) * 2002-03-06 2003-09-18 Schilling Amanda S. Application of germination solution improved efficacy of biological decontamination
US6656919B1 (en) * 2002-01-11 2003-12-02 Clarence L. Baugh Method and a product for the rapid decontamination and sterilization of bacterial endospores
US20040058878A1 (en) * 2002-01-18 2004-03-25 Walker Edward B. Antimicrobial and sporicidal composition
US20120148751A1 (en) * 2010-12-14 2012-06-14 Ecolab Usa Inc. Wear resistant antimicrobial compositions and methods of use

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6656919B1 (en) * 2002-01-11 2003-12-02 Clarence L. Baugh Method and a product for the rapid decontamination and sterilization of bacterial endospores
US20040058878A1 (en) * 2002-01-18 2004-03-25 Walker Edward B. Antimicrobial and sporicidal composition
US20030175318A1 (en) * 2002-03-06 2003-09-18 Schilling Amanda S. Application of germination solution improved efficacy of biological decontamination
US20120148751A1 (en) * 2010-12-14 2012-06-14 Ecolab Usa Inc. Wear resistant antimicrobial compositions and methods of use

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