JP5148998B2 - Detergent for temperature chamber - Google Patents

Description

  The present invention relates to a novel detergent having antiseptic, antifungal and antialgal effects added to a reaction thermostat using water as a medium in scientific instruments, particularly to a thermostat in an automatic analyzer.

  In general, in the field of clinical chemistry, measurement of physiologically active substances such as enzymes, lipids, and proteins in biological samples such as serum, urine, and tissue fluid is widely performed for the purpose of diagnosing diseases and grasping disease states.

  The automatic analyzer has various features such as rapidness, high efficiency and high accuracy, simple operation, small amount of sample / reagent, and labor saving. Widely used. The measurement is usually performed in the order of sample collection, reagent addition, mixing, heating, colorimetry (absorbance measurement), and calculation. Among these, there are an air bath (air bath) method and a water bath or oil bath method as heating, but the most common type is a water bath as a thermostatic bath. The reaction temperature is usually 50 ° C. or lower, but the most common is 37 ° C. Absorbance measurement includes a method in which a reaction solution is sucked into a cell from a reaction vessel and a method in which the reaction vessel is directly measured as a measurement cell, but the latter method is mainly used at present. In this method, when a water bath is used as a thermostatic bath, the light emitted from the light source installed outside the bath passes through the thermostatic bath, and then passes through a reaction vessel in the thermostatic bath with a detector installed on the opposite side of the bath. This is done by being detected. That is, the reaction vessel is also measured directly as a reaction cell. The wavelength used for the measurement is usually 300 to 900 nm.

  By the way, the water in the thermostat of the automatic analyzer is usually exchanged once to several times a day, but bubbles often adhere to the outer wall of the reaction vessel during the water exchange. For this purpose, a small amount of detergent is usually added. The detergent used for this purpose is usually composed mainly of various surfactants, added with chelating agents, pH adjusters, preservatives, etc., and has low foaming properties. The concentration of the detergent is usually 0.05 to 2.0 w / w%. However, the water in the thermostatic bath that uses the water in the thermostatic bath containing such a detergent becomes a nutrient source when used for a long time, and promotes the generation of algae and the growth of microorganisms (bacteria etc.). (Even if preservatives are added, the effect is not recognized so much) As a result, the cause of large errors in absorbance measurement, such as adhesion of algae to the reaction vessel and turbidity of water in the thermostatic bath, etc. It was. For this reason, when using the apparatus, the actual situation is that much effort is required for accuracy management and maintenance management, such as frequently monitoring the inside of the tank and periodically cleaning the inside of the tank. As described above, the addition of the detergent to the thermostatic bath is necessary and effective, but daily maintenance management must be sufficiently performed. Therefore, improvement of this point has been strongly demanded.

  The inventors of the present invention believe that the generation of algae and the growth of microorganisms (bacteria, etc.) are caused by a constant temperature bath of a preservative that exists as a component of the normal detergent concentration (0.05 to 2.0 v / v%). As a result of diligent research, we focused on triazine derivatives as preservatives that are effective when added in small amounts, that is, effective at low concentrations. It has been found that the use of a detergent containing an activator as a component can prevent the generation of algae and the growth of microorganisms (bacteria and the like) in a thermostatic bath, and a patent application has already been filed (Patent Document 1).

  However, when a detergent having this composition was used, it was possible to prevent the generation of algae and the growth of microorganisms (bacteria, etc.) in the thermostatic bath for a long period of time, but the detergent stock solution was kept at a high temperature. When stored, some of these components decompose over time, resulting in a substance that absorbs at the wavelength used for measurement (300-900 nm), causing a large error in absorbance measurement. It turns out that there is a possibility.

Therefore, as a result of further diligent research, the present inventors have further added a detergent that combines a triazine derivative as a preservative with a low effective concentration and a surfactant to the following general formula [A].

(Wherein R ₉ , R ₁₀ , R ₁₁ and R ₁₂ each independently represents a hydrogen atom, a methyl group or a hydroxymethyl group, and n represents an integer of 1 to 5). In addition to preventing the growth of algae and the growth of microorganisms (bacteria, etc.) in the thermostatic bath, when the detergent itself is stored at high temperatures, some of its components decompose and measure. It has been found that generation of a substance having absorption at a wavelength (300 to 900 nm) used in the present invention can be suppressed, and a patent application has already been filed (Patent Document 2).

  On the other hand, there are voices for various pretreatments to prevent nosocomial infections caused by various bacteria (for example, Pseudomonas aeruginosa, Staphylococcus aureus, etc.). Although it has never been reported, there is also a desire to develop a cleaner with a stronger bactericidal effect than conventional cleaners, due to the idea that prior prevention is also necessary.

Japanese Patent Laid-Open No. 1-40599 JP-A-1-319407

  The present invention has been made in view of the situation as described above, and is a detergent added to a reaction thermostatic bath using water as a medium in scientific instruments, particularly a thermostatic bath in an automatic analyzer, It has antiseptic, antifungal, and algal control effects over a long period of time, and does not produce a substance that absorbs at wavelengths (300 to 900 nm) used for measurement due to decomposition of some of its constituent components (or The purpose is to provide a new detergent that has a higher sterilizing effect (has a sterilizing effect in a short time) than conventional ones and can meet the demand for prevention and prevention of nosocomial infections. .

  The present invention has been made for the purpose of solving the above-described problems, and has the following configuration.

(1) A quaternary ammonium salt represented by the following general formula [1]

(Wherein R _{1 to} R ₄ each independently represents an alkyl group, provided that at least one of R _{1 to} R ₄ is an alkyl group having 8 to 18 carbon atoms, and at least one is carbon. the number is 1 to 3 lower alkyl groups .X ^- represents an anion derived from a carboxylic acid having 2 or more carbon atoms).
Or the antiseptic | preservative which has amphoteric surface active ability, and the detergent for thermostats containing surfactant.
(2) A method for cleaning a thermostat, comprising adding the detergent described in (1) to the thermostat.

  That is, the conventional detergent for a thermostatic bath was excellent in antibacterial effect, but there was also a demand for prevention / prevention of nosocomial infection as described above. Therefore, the present inventors have found out that among the widely used antibacterial agents and bactericides, the ones that can achieve the above-mentioned purpose are developed for the purpose of developing a novel thermostatic bath detergent that is also excellent in bactericidal effect. , Earnest research.

  In an automatic analyzer equipped with a thermostat, a sample and a reagent are mixed, heated in the thermostat, and then the absorbance is measured as it is using the reaction vessel as a measurement cell. Therefore, it is necessary that the detergent for the thermostatic bath has no absorption in the measurement wavelength range (300 to 900 nm) of the automatic analyzer. Therefore, for example, chlorhexidine, chlorhexidine gluconate, etc., which are widely used among antibacterial agents and fungicides, absorb at 300 to 400 nm, and cetylpyridinium halide, etc., has an aqueous solution that is pale yellow to yellow, and this coloring is UV. None of them were suitable for use in a thermostatic bath detergent because they impeded absorption.

  Moreover, since serum, enzymes, and the like are used in the measurement using an automatic analyzer, there is a risk that these proteins will contaminate the water in the thermostatic chamber. Therefore, even if these proteins are mixed in the water in the thermostatic chamber, it is necessary to suppress the sterilization of bacteria or the propagation thereof, and in this respect, benzalkonium chloride, benzethonium chloride, etc. In the presence of protein, a decrease in antibacterial activity and the like was observed, and dissatisfaction remained for use in a thermostatic bath detergent.

  Furthermore, as described above, in order to prevent or prevent nosocomial infections, it is desirable to have a bactericidal activity against Pseudomonas aeruginosa and the like.

Accordingly, the present inventors have further conducted extensive studies, and as a result, the following general formula [I]

(Wherein R _{1 to} R ₄ each independently represents an alkyl group, provided that at least one of R _{1 to} R ₄ is an alkyl group having 8 to 18 carbon atoms, and at least one is carbon. A quaternary ammonium salt represented by the formula (1) ^-( C3), and a preservative having an amphoteric surface active ability. The present invention was completed by finding that all the points were satisfied, and that the detergent for a thermostatic bath containing them could achieve all of the objects of the present application.

  The present invention prevents the generation of microorganisms (bacteria etc.) generated in the water in the thermostat bath and the accompanying decrease in measurement accuracy in scientific instruments having a thermostat bath using water as a medium, particularly in an automatic analyzer. In addition, it prevents the generation and adhesion of bubbles on the outer wall of the reaction vessel in the thermostat, and it itself absorbs at the measurement wavelength (300 to 900 nm) due to decomposition of some of its constituent components when storing the stock solution This is to provide a thermostatic bath cleaner that has a higher sterilizing effect than the conventional one. The present invention provides an excellent temperature-controlled bath detergent that can take advantage of the advantages of quickness, high efficiency, high accuracy, and ease of operation.

The detergent for a thermostatic bath according to the present invention is a quaternary ammonium salt represented by the following general formula [I].

(Wherein R _{1 to} R ₄ each independently represents an alkyl group, provided that at least one of R _{1 to} R ₄ is an alkyl group having 8 to 18 carbon atoms, and at least one is carbon. the number is 1 to 3 lower alkyl groups .X ^- are those containing a preservative having) or / and amphoteric surface activity represents an anion derived from a carboxylic acid having 2 or more carbon atoms, a surfactant..

  In the thermostatic bath detergent according to the present invention, the quaternary ammonium salt represented by the general formula [1] or / and the preservative having amphoteric surface activity and the surfactant are usually dissolved in a suitable solvent. However, as the solvent, water is particularly preferable.

The alkyl group represented by R _{1 to} R ₄ of the quaternary ammonium salt represented by the general formula [1] used in the thermostatic bath detergent according to the present invention may be linear or branched. It may be cyclic and usually includes those having 1 to 20 carbon atoms, preferably 1 to 12 carbon atoms. Specifically, for example, methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, tert-butyl group, n-pentyl group, isopentyl group, sec-pentyl group, tert-pentyl group, neopentyl group, n-hexyl group, isohexyl group, sec-hexyl group, tert-hexyl group, 3 -Methylpentyl group, 2-methylpentyl group, 1,2-dimethylbutyl group, n-heptyl group, isoheptyl group, sec-heptyl group, n-octyl group, isooctyl group, sec-octyl group, n-nonyl group, n-decyl group n-undecyl group, n-dodecyl group, n-tridecyl group, n-tetradecyl group, n-pentadecyl group, n-hexadecyl group, n-heptadecyl group, n-octadecyl group, n-nonadecyl group, n-icosyl group, Cyclopropyl group, cyclopentyl group, cyclohexyl group, cycloheptyl group, cyclooctyl group, cyclodecyl group, cycloundecyl group, cyclododecyl group, cyclotetradecyl group, cyclooctadecyl group, cycloicosyl group, bicyclo [2.1.0] pentyl group , Bicyclo [3.2.1] octyl group, bicyclo [5.2.0] nonyl group, tricyclo [5.3.1.1] dodecyl group, perhydroanthryl group, spiro [3.4] octyl group, spiro [4.5] decyl group, etc. Of these, linear ones are preferred. However, at least one of R _{1 to} R ₄ is an alkyl group having 8 to 18 carbon atoms, and at least one is a lower alkyl group having 1 to 3 carbon atoms. Specific examples of each group are appropriately selected from the alkyl groups exemplified above. Preferred examples of the alkyl group having 8 to 18 carbon atoms include n-decyl group, n-undecyl group, and n-dodecyl group. Among them, a decyl group is preferable. Preferable specific examples of the lower alkyl group having 1 to 3 carbon atoms include a methyl group, an ethyl group, a propyl group, and the like, and among them, a methyl group is preferable.

In the general formula [1], the anion derived from a carboxylic acid having 2 or more carbon atoms represented by X ⁻ is represented by the general formula [2].

(Wherein R ₅ represents a hydrogen atom or a monovalent hydrocarbon group which may have a substituent) or a general formula [3]

(Wherein R ₆ represents a bond or a divalent hydrocarbon group which may have a substituent).

In the general formula [2], the hydrocarbon group of the monovalent hydrocarbon group which may have a substituent represented by R ₅ includes an aliphatic hydrocarbon group, an aromatic hydrocarbon group and an aromatic group. An aliphatic hydrocarbon group is mentioned, You may have a sulfur atom in the chain | strand of those groups.

  Examples of the aliphatic hydrocarbon group include an alkyl group and an alkenyl group.

  The alkyl group may be linear, branched or cyclic, and usually has 1 to 20 carbon atoms, preferably 1 to 12 carbon atoms. Specifically, for example, a methyl group, an ethyl group, n- Propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, tert-butyl group, n-pentyl group, isopentyl group, sec-pentyl group, tert-pentyl group, neopentyl group, n-hexyl group, Isohexyl group, 3-methylpentyl group, 2-methylpentyl group, 1,2-dimethylbutyl group, n-heptyl group, isoheptyl group, sec-heptyl group, n-octyl group, isooctyl group, sec-octyl group, n -Nonyl, n-decyl, n-undecyl, n-dodecyl, n-tridecyl, n-tetradecyl, n-pentadecyl, n-hexadecyl, n-heptadecyl, n-octadecyl, n - Nadecyl group, n-icosyl group, cyclopropyl group, cyclopentyl group, cyclohexyl group, cycloheptyl group, cyclooctyl group, cyclodecyl group, cycloundecyl group, cyclododecyl group, cyclotetradecyl group, cyclooctadecyl group, cycloicosyl group, etc. Is mentioned.

  The alkenyl group may be linear, branched or cyclic, and usually has 2 to 20 carbon atoms, preferably 2 to 12 carbon atoms. Specific examples thereof include vinyl groups, allyl groups, 1- Propenyl group, isopropenyl group, 3-butenyl group, 2-butenyl group, 1-butenyl group, 1,3-butadienyl group, 4-pentenyl group, 3-pentenyl group, 2-pentenyl group, 1-pentenyl group, 1 , 3-pentadienyl group, 2,4-pentadienyl group, 1,1-dimethyl-2-propenyl group, 1-ethyl-2-propenyl group, 1,2-dimethyl-1-propenyl group, 1-methyl-1- Butenyl group, 5-hexenyl group, 4-hexenyl group, 3-hexenyl group, 2-hexenyl group, 1-hexenyl group, 1-methyl-1-hexenyl group, 2-methyl-2-hexenyl group, 3-methyl- 1,3-hexadienyl group, 1-heptenyl group, 2-octenyl group, 3-nonenyl group, 4-decenyl group, 1-unde Nyl group, 2-dodecenyl group, 3-tridecenyl group, 4-tetradecenyl group, 5-pentadecenyl group, 6-hexadecenyl group, 7-heptadecenyl group, 3-octadecenyl group, 1-nonadecenyl group, 2-icosenyl group, 1- Cyclopropenyl group, 2-cyclopentenyl group, 2,4-cyclopentanedienyl group, 1-cyclohexenyl group, 2-cyclohexenyl group, 3-cyclohexenyl group, 2-cycloheptenyl group, 2-cyclononenyl group, 3- Examples include a cyclododecenyl group, a 3-cyclopentadecenyl group, a 2-cyclooctadecenyl group, and a 2-cycloicosenyl group.

  As the aromatic hydrocarbon group, an aryl group is preferable, and specific examples thereof usually include those having 6 to 20 carbon atoms, preferably 6 to 14 carbon atoms. Specific examples include a phenyl group, a naphthyl group, an anthryl group, Examples thereof include 1-pyrenyl group and perylenyl group.

  As the araliphatic hydrocarbon group, an aralkyl group is preferable, and specifically, an alkyl group in which a hydrogen atom is substituted with an aromatic ring, usually having 7 to 13 carbon atoms, preferably 7 to 10 carbon atoms. Specific examples include a benzyl group, a phenethyl group, a phenylpropyl group, a phenylbutyl group, a phenylpentyl group, a phenylhexyl group, a phenyl-1-methylhexyl group, and a phenyl-3-cyclopentyl group.

Examples of the substituent of the monovalent hydrocarbon group which may have a substituent represented by R ₅ include halogen atoms such as a chlorine atom, a fluorine atom, a bromine atom and an iodine atom, such as a methyl group and an ethyl group. , N-propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, tert-butyl group and other lower alkyl groups such as fluoromethyl group, difluoromethyl group, trifluoromethyl group, bromomethyl group, Bromomethyl group, chloromethyl group, dichloromethyl group, trichloromethyl group, trifluoroethyl group, tribromoethyl group, trichloroethyl group, pentafluoroethyl group, pentabromoethyl group, pentachloroethyl group, heptafluoropropyl group, Halo lower alkyl groups such as heptabromopropyl and heptachloropropyl, eg Examples include lower alkoxyl groups such as methoxy group, ethoxy group, propoxy group, butoxy group, amino group, nitro group, oxo group, hydroxyl group, heterocyclic group, aldehyde group, etc. Among them, halogen atom having electron withdrawing property , A halo lower alkyl group, an alkyl group and a nitro group are preferred.

  Examples of the heterocyclic group include 5- to 6-membered ones having 1 to 3 nitrogen atoms, sulfur atoms and / or oxygen atoms as isomer atoms. Specific examples include 2-tetrahydrofuryl group and 2-tetrahydro Heterocyclic aliphatic groups such as thienyl group, 1-pyrrolidinyl group, 2-pyrrolidinyl group, 4-piperidinyl group, 2-morpholinyl group, such as 2-furyl group, 2-thienyl group, 1-pyrrolyl group, 2-pyridyl group Group, 3-pyridyl group, isobenzofuranyl group, chromenyl group, 2H-pyrrolyl group, imidazolyl group, pyrazolyl group, 5-pyrazolyl group, indolizinyl group, isoindolyl group, 3H-indolyl group, indolyl group, 3-indolyl group , 1H-indazolyl group, heterocyclic aromatic group such as purinyl group, and the like.

Specific examples of the compound represented by the general formula [2] include, for example, acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid, pivalic acid, pentanoic acid, hexanoic acid, heptanoic acid, octanoic acid, and nonanoic acid. , decanoic acid, undecanoic acid, lauric acid, myristic acid, pentadecanoic acid, palmitic acid, heptadecanoic acid, stearic acid, nonadecanoic acid, aliphatic saturated monocarboxylic acids such as eicosane acids such as aliphatic cyclic mono- and cyclohexyl carboxylic acid Carboxylic acids such as fluoroacetic acid, trifluoroacetic acid, chloroacetic acid, dichloroacetic acid, trichloroacetic acid, bromoacetic acid, iodoacetic acid, perfluoropropionic acid, perchloroheptanoic acid, perfluoroheptanoic acid, perfluorooctanoic acid, perfluorodecanoic acid , perfluoro dodecanoic acid, perfluoro Eiko Phosphate, halogenated alkyl monocarboxylic acids such as perfluoro-tetra co Sa phosphate, such as acrylic acid, propiolic acid, methacrylic acid, crotonic acid, isocrotonic acid, 4-hexenoic acid, oleic acid, aliphatic, such as elaidic acid- Saturated monocarboxylic acids, for example alicyclic monocarboxylic acids such as camphoric acid and adamantanoic acid, aromatic monocarboxylic acids such as benzoic acid, naphthoic acid and anthracene carboxylic acid, alkyl aromatic monocarboxylic acids such as toluic acid, For example, fluorobenzoic acid, chlorobenzoic acid, bromobenzoic acid, difluorobenzoic acid, dichlorobenzoic acid, dibromobenzoic acid, trifluorobenzoic acid, trichlorobenzoic acid, tribromobenzoic acid, tetrafluorobenzoic acid, tetrachlorobenzoic acid, tetrabromobenzoic acid Acid, pentafluorobenzoic acid, penta Halogenated aromatic monocarboxylic acids such as chlorobenzoic acid and pentabromobenzoic acid, for example, halogenated alkyl aromatic monocarboxylic acids such as trifluoromethylbenzoic acid, trichloromethylbenzoic acid, bis (trifluoromethyl) benzoic acid, Halogenated alkoxyaromatic monocarboxylic acids such as fluoromethoxybenzoic acid and trichloromethoxybenzoic acid, nitroaromatic monocarboxylic acids such as trinitrobenzoic acid, aralkyl monocarboxylic acids such as 2-phenylpropanoic acid, such as hydroatropic acid, etc. Aralkyl acids such as, for example, arylalkenyl acids such as cinnamic acid and atropic acid, hydroxyaliphatic monocarboxylic acids such as glycolic acid, lactic acid and glyceric acid, aromatic hydroxyalkyl monocarboxylic acids such as benzylic acid and tropic acid, For example, hydroxyaromatic monocarboxylic acids such as salicylic acid, protocatechuic acid, gallic acid, anisic acid, vanillic acid, aliphatic ketone monocarboxylic acids such as pyruvic acid, acetoacetic acid, such as alanine, arginine, asparagine, cysteine, glutamine, glycine , Histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, and other amino acids such as nicotinic acid, isonicotinic acid, furancarboxylic acid, thiophenecarboxylic acid, 1-pyrrolecarboxylic acid, etc. And heterocyclic monocarboxylic acids such as p-formylphenylacetic acid and 6- (2-naphthyl) hexanoic acid.

In the general formula [3], the divalent hydrocarbon group of the divalent hydrocarbon group which may have a substituent represented by R ₆ includes a divalent aliphatic hydrocarbon group, two Valent aromatic hydrocarbon group and divalent araliphatic hydrocarbon group.

  Examples of the divalent aliphatic hydrocarbon group include an alkylene group and an alkenylene group.

The alkylene group may be linear, branched or cyclic, and usually has 1 to 10 carbon atoms , preferably 1 to 6 carbon atoms . Specific examples include methylene group, ethylene group and trimethylene group. , Propylene group, tetramethylene group, butylene group, 2-methylpropylene group, pentamethylene group, pentylene group, 2-methyltetramethylene group, 2,2-dimethyltrimethylene group, 2-ethyltrimethylene group, hexamethylene group Hexylene group, 2-methylpentamethylene group, 3-methylpentamethylene group, heptamethylene group, heptylene group, octamethylene group, octylene group, 2-ethylhexylene group, nonamethylene group, nonylene group, decamethylene group, decylene group , Cyclopropylene group, cyclopentylene group, cyclohexylene group, cycloheptylene Group, cyclooctylene group, cyclononylene group, a cyclodecylene group, and the like.

The alkenylene group may be linear, branched or cyclic, and usually has 2 to 10 carbon atoms , preferably 2 to 6 carbon atoms . Specific examples include vinylene groups, propenylene groups, 1- Butenylene group, 2-butenylene group, 1-pentenylene group, 2-pentenylene group, 2-methyl-1-propenylene group, 2-methyl-1-butenylene group, 1-methyl-1-butenylene group, 1-hexenylene group, 2-hexenylene group, 3-hexenylene group, 1-heptenylene group, 2-heptenylene group, 3-heptenylene group, 1-octenylene group, 2-octenylene group, 3-octenylene group, 4-propyl-2-pentenylene group, 1 -Nonenylene group, 2-nonenylene group, 1-decenylene group, 4-cyclopentene-1,3-ylene group, 3-cyclohexene-1, 2- Ren group, and the like.

  Examples of the divalent aromatic hydrocarbon group include an arylene group.

  Examples of the arylene group usually include those having 6 to 10 carbon atoms. Specific examples include an o-phenylene group, an m-phenylene group, a p-phenylene group, a 1,5-naphthylene group, and a 1,8-naphthylene. Group, 2,7-naphthylene group, 2,6-naphthylene group and the like.

Examples of the divalent araliphatic group usually include those having 7 to 12 carbon atoms. Specifically, for example, —CH ₂ —C ₆ H ₄ —, —C ₂ H ₄ —C ₆ H ₄ —, _{-CH (CH 3) -C 6 H} 4 -, - CH 2 -C 6 H 4 -CH 2 -, - CH (CH 3) -C 6 H 4 -C 2 H 4 -, - C 3 H 6 - _{C 6 H 4 -CH 2 -,} - C 3 H 6 -C 6 H 4 -C 2 H 4 -, - C 3 H 6 -C 6 H 4 -C 3 H 6 -, - CH 2 CH (CH 3 ) -C ₆ H ₄ -C ₂ H _4- and the like.

Examples of the substituent of the divalent hydrocarbon group which may have a substituent represented by R ₆ include the substituent in the monovalent hydrocarbon group which may have the substituent of R ₅ above. Similar groups are mentioned.

  Specific examples of the compound represented by the general formula [3] include aliphatic saturated dicarboxylic acids such as oxalic acid, malonic acid, succinic acid, glutaric acid, adipic acid, pimelic acid, suberic acid, azelaic acid and sebacic acid, For example, aliphatic unsaturated dicarboxylic acids such as 4-propyl-2-pentenedioic acid, maleic acid, fumaric acid, citraconic acid, mesaconic acid, such as phthalic acid, isophthalic acid, terephthalic acid, 1,5-naphthalenedicarboxylic acid, etc. Aromatic dicarboxylic acids, for example, hydroxyaliphatic dicarboxylic acids such as tartronic acid, malic acid, tartaric acid, amino acids such as aspartic acid, cystine, glutamic acid, heterocyclic dicarboxylic acids such as 2,3-quinoline diacetic acid, etc. It is done.

  The quaternary ammonium salt that can be used for the purpose of the present invention has almost no absorption at the measurement wavelength (300 to 900 nm) at the concentration used, and is compatible with water or / and a surfactant, Precipitation with other components in the cleaning agent, no turbidity, no corrosion of glass, plastic, metal, etc., stable quality for a long time, generation of algae at low effective concentration, microorganisms (bacteria etc. ), And the like.

  Specific examples thereof include dialkyldimethylammonium adipate and dialkyldimethylammonium propionate. Dialkyldimethylammonium adipate is particularly preferable, and among them didecyldimethylammonium adipate is preferable.

  These may be used as they are on the market. For example, didecyldimethylammonium adipate is commercially available under the name of osmolin DA-50 (trade name of Sanyo Chemical Industries, Ltd.).

The quaternary ammonium salt represented by the general formula [1] may be used alone or in combination of two or more. The amount used may be a concentration that prevents the generation of algae and the growth of microorganisms (bacteria, etc.) and hardly affects the measurement, and the concentration in a constant temperature bath is generally 0.001 to 0.1 w / It is added to the detergent so as to be about w%, preferably 0.005 to 0.05 w / w%. The concentration in the detergent may be determined in consideration of how many times it is diluted when added to water in a thermostatic bath, but is usually about 1 to 10 w / w%, preferably 3 to It is used alone or in combination of two or more so as to be about 8 w / w%.

  The preservative having amphoteric surface activity used in the detergent for a thermostatic bath according to the present invention is the same as the quaternary ammonium salt used in the present invention at the measurement concentration (300 to 900 nm). Has no absorption, is compatible with water or / and surfactant, does not precipitate with other components in the detergent, does not cause turbidity, does not corrode glass, plastic, metal, etc. It is desirable to have properties such as stable quality for a long period of time and the ability to prevent the development of algae and the growth of microorganisms (bacteria etc.) at a low effective concentration.

の構造を持つN−ビス(3−アミノプロピル)ドデシルアミン、3−アミノプロピル−ドデシルプロパンジアミン、1,3−プロパンジアミン−3−アミノプロピル−N−ドデシルが、特にN-ビス（３−アミノプロピル）ドデシルアミンが好ましい。Specific examples thereof include, for example, N-bis (3-aminopropyl) dodecylamine, 3-aminopropyl-dodecylpropanediamine, 1,3-propanediamine-N-3-aminopropyl-N-dodecyl, alkylaminoglycine, Examples include alkyldiaminoethylglycine hydrochloride, alkylaminoethylaminoethylglycine, alkylaminopropylaminoacetic acid, alkylpolyaminoethylglycine, and the like.

N-bis (3-aminopropyl) dodecylamine, 3-aminopropyl-dodecylpropanediamine, 1,3-propanediamine-3-aminopropyl-N-dodecyl having the structure of N-bis (3-amino Propyl) dodecylamine is preferred.

  These may be used as they are on the market. For example, N-bis (3-aminopropyl) dodecylamine, Lonzabak 12.100 (Lonzabac 12.100, trade name of Lonza Japan Co., Ltd.) and the like are sold.

  The preservatives may be used alone or in combination of two or more. The amount used may be a concentration that prevents the generation of algae and the growth of microorganisms (bacteria, etc.) and hardly affects the measurement, and the concentration in a constant temperature bath is generally 0.0005 to 0.02 w / It is added to the detergent so as to be about w%, preferably 0.001 to 0.01 w / w%. The concentration in the detergent may be determined in consideration of how many times it is diluted when added to water in a thermostatic bath, but is usually about 0.05 to 10 w / w%, preferably 1 to It is used alone or in combination of two or more so as to be about 5 w / w%.

  As the surfactant used in the detergent for the thermostatic bath according to the present invention, any surfactant can be used as long as it has no effect on measurement and can prevent bubbles from adhering to the reaction vessel. Quaternary ammonium salt represented by the general formula [1] according to the present invention, free from water-insoluble substances, low foaming property, high cloud point and clear at reaction temperature (37 ° C). Or it does not cause reaction and precipitation with preservatives having amphoteric surface activity, there is almost no absorption in the range of 300-900 nm, glass, metal, plastic, etc. constituting the thermostat and reaction vessel of automatic analyzer There is no particular limitation as long as it has properties such as being unaffected and being always stable in quality, free of danger and easy to handle.

  For example, any of nonionic surfactants, cationic surfactants, anionic surfactants, and amphoteric surfactants can be used. Examples of nonionic surfactants include polyoxyethylene alkyls. Ether, polyoxyethylene alkyl phenyl ether, polyoxyethylene fatty acid ester, polyoxyethylene sorbitan fatty acid ester, sorbitan fatty acid ester, polyoxyethylene sorbitol fatty acid ester, polyoxyethylene alkylamine, glycerin fatty acid ester, fatty acid alkanolamide, sucrose fatty acid Examples of the cationic surfactant include aliphatic amine salts and aliphatic quaternary ammonium salts. Examples of the anionic surfactant include carboxylate, sulfonate, and sulfate. salt, Phosphate ester salts and the like, and the amphoteric surface active agent, such as carboxymethyl betaines, sulfobetaines, glycines, alanine compounds, derivatives of 2-alkyl imidazolines, amine oxides, and the like.

  Furthermore, specific examples of polyoxyethylene alkyl ether which is a nonionic surfactant include, for example, polyoxyethylene cetyl ether, polyoxyethylene stearyl ether, polyoxyethylene oleyl ether, and the like. Specific examples of ethers include, for example, polyoxyethylene octyl phenyl ether, polyoxyethylene nonyl phenyl ether, and the like, and specific examples of polyoxyethylene fatty acid esters include, for example, polyoxyethylene glycol monolaurate, polyoxyethylene glycol Examples include monostearate, polyoxyethylene glycol distearate, polyoxyethylene glycol monooleate, and polyoxyethylene sorbitan fatty acid ester. Specific examples include polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monopalmitate, polyoxyethylene sorbitan monostearate, polyoxyethylene sorbitan tristearate, polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan Examples of sorbitan fatty acid esters include sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan distearate, sorbitan tristearate, sorbitan monooleate, sorbitan trioleate, and the like. Sorbitan sesquioleate and the like, and specific examples of polyoxyethylene sorbitol fatty acid ester include, for example, tetraoleic acid Specific examples of polyoxyethylene alkylamine include, for example, polyoxyethylene laurylamine, polyoxyethylene stearylamine, and specific examples of glycerin fatty acid ester include, for example, stearic acid monoglyceride. Specific examples of fatty acid alkanolamides include, for example, lauric acid diethanolamide, and specific examples of sucrose fatty acid esters include, for example, sucrose palmitate, sucrose stearate, and the like. Etc.

  Examples of the aliphatic amine salt that is a cationic surfactant include higher aliphatic amines such as monolaurylamine, monostearylamine, distearylamine, and tristearylamine, and inorganic acids such as hydrochloric acid and sulfuric acid, or acetic acid and lactic acid. And salts with lower carboxylic acids such as citric acid, and the like. Specific examples include laurylamine acetate and stearylamine acetate. Aliphatic quaternary ammonium salts include lauryltrimethylammonium and stearyl. Examples include salts of higher aliphatic ammonium such as trimethylammonium, cetyltrimethylammonium, didecyldimethylammonium, benzyldimethyltetradecylammonium and the like with chlorine, bromine and the like. Specific examples include lauryltrimethylammonium chloride, stearyltrimethylammonium. Umukuroraido, cetyl trimethyl ammonium chloride, didecyl dimethyl ammonium chloride, benzyl dimethyl tetradecyl ammonium chloride and the like.

  Examples of carboxylates that are anionic surfactants include salts with higher fatty acids such as lauric acid, myristic acid, palmitic acid, stearic acid, and oleic acid, and alkali metals such as sodium and potassium. Specific examples include potassium oleate, sodium lauroyl sarcosine, N-myristoyl-N-methyl-β-alanine sodium, sodium polyoxyethylene lauryl ether acetate, and the sulfonates include lauryl benzene sulfonic acid. And alkylbenzene sulfonic acid such as dipropyl naphthalene sulfonic acid, dibutyl naphthalene sulfonic acid, and the like, and salts of sulfosuccinic acid such as dioctyl sulfosuccinic acid and sodium, and the like. Sodium acid, Examples include sodium propyl naphthalene sulfonate, sodium dibutyl naphthalene sulfonate, sodium dioctyl sulfosuccinate, etc., and sulfate esters include higher alcohol sulfates such as lauryl sulfate, and polyoxyethylene alkyls such as polyoxyethylene lauryl ether sulfate. Examples thereof include salts of esters such as ether sulfate and sodium, ammonium and the like. Specifically, higher alcohol sulfate esters such as sodium lauryl sulfate and ammonium lauryl sulfate, and polyoxyethylene alkyl such as sodium polyoxyethylene lauryl ether sulfate. Ether sulfate esters and the like. Examples of phosphate ester salts include monostearyl phosphate, monolauryl phosphate, polyoxyethylene. Examples thereof include salts of lauryl ether phosphate and the like with alkali metals such as sodium and potassium. Specific examples include sodium monostearyl phosphate, sodium monolauryl phosphate, and potassium polyoxyethylene lauryl ether phosphate. Can be mentioned.

  Specific examples of carboxybetaines that are amphoteric surfactants include lauric acid amidopropyl betaine, lauryldimethylaminoacetic acid betaine, N-lauroyl-N′-carboxymethyl-N′-hydroxyethylethylenediamine sodium, and the like, Specific examples of sulfobetaines include lauric acid amidopropyl hydroxysulfobetaine, specific examples of glycines include sodium lauryldiaminoethylglycine, and specific examples of 2-alkylimidazoline derivatives Include 2-alkyl-N-carboxymethyl-N-hydroxyethylimidazolinium betaines such as 2-lauroyl-N-carboxymethyl-N-hydroxyethylimidazolinium betaine, and specific examples of amine oxides Lauryldimethylamine Kisaido, and the like.

  Specific examples of these surfactants include polyoxyethylene alkyl ether and sodium lauroyl sarcosine, and it is more preferable to use these in combination.

  Moreover, these surfactants may be used as they are. For example, polyoxyethylene alkyl ether is sold under the name of BT-9 (trade name of Nikko Chemicals Co., Ltd.), and lauroyl sarcosine sodium is sold under the name of sarcosinate LN (trade name of Nikko Chemicals Co., Ltd.). .

  These surfactants may be used alone or in combination of two or more. In addition, the amount used may be a concentration that does not easily affect the measurement, and the concentration in the constant temperature bath is generally about 0.005 to 1.0 w / w%, preferably 0.01 to 0.5 w / w%. In the detergent. The concentration in the detergent may be determined in consideration of how many times it is diluted when added to water in a thermostatic bath, but is usually about 1 to 20 w / w%, preferably 5 to It is used alone or in combination of two or more so as to be about 10 w / w%.

  The detergent for the thermostatic bath according to the present invention only needs to contain the quaternary ammonium salt according to the present invention or the preservative having amphoteric surface active ability and the surfactant according to the present invention at a predetermined concentration. Quaternary ammonium salts are excellent in antibacterial activity, and preservatives having amphoteric surface activity have higher bactericidal activity, so it is more preferable that they coexist.

In addition, the detergent for thermostats which concerns on this invention is mainly added in the thermostat water tank of an automatic analyzer, and is usually added in a water tank via a small diameter plastic tube. For this reason, when the viscosity of the cleaning agent itself becomes too high or becomes cloudy, a predetermined amount of cleaning agent cannot be added to the constant temperature bath, or the plastic tube for transfer is clogged. There is a high possibility that problems will occur in practice. Accordingly, it is desirable that the cleaning agent itself be clear and have a viscosity of less than 6 centistokes (cSt, mm ² / s in SI unit system ).

  In addition, as long as the detergent for the thermostatic bath according to the present invention does not impair the object of the present invention, a chelating agent, a pH adjuster, for example, a stabilizer such as β-thiodiglycol, and the like are added as necessary. It is also optional to do.

  The chelating agent contained as needed in the thermostatic bath detergent according to the present invention is not particularly limited as long as it forms a metal impurity and a complex compound. For example, one or more carboxyl groups in the molecule , Compounds having one or more phosphonic acid groups in the molecule, N-substituted amino acids, condensed phosphoric acids, and ammonium salts or alkali metal salts thereof.

  As the compound having one or more carboxyl groups in the molecule, nitrogen-containing polycarboxylic acids having 1 to 4 nitrogen atoms and 2 to 6 carboxyl groups in the molecule are preferable. Specifically, for example, hydroxyethyl Alkyliminopolycarboxylic acids which may have a hydroxy group such as iminodiacetic acid [HIDA] and iminodiacetic acid [IDA], for example, nitrilopolycarboxylic acids such as nitrilotriacetic acid [NTA] and nitrilotripropionic acid [NTP] Acids such as ethylenediaminetetraacetic acid [EDTA], ethylenediaminediacetic acid [EDDA], ethylenediaminedipropionic acid dihydrochloride [EDDP], hydroxyethylethylenediaminetriacetic acid [EDTA-OH], 1,6-hexamethylenediamine-N, N , N ', N'-tetraacetic acid [HDTA], triethylenetetramine hexaacetic acid [TTHA] Hydroxyalkyl groups such as diethylenetriamine-N, N, N ', N' ', N' '-pentaacetic acid [DTPA], N, N-bis (2-hydroxybenzyl) ethylenediamine-N, N-diacetic acid [HBED] , Mono or polyalkylene polyamine polycarboxylic acid optionally having a hydroxyaryl group or a hydroxyaralkyl group, for example, diaminopropanetetraacetic acid [Methyl-EDTA], trans-1,2-diaminocyclohexane-N, N, N ′ , N'-tetraacetic acid [CyDTA] and other polyaminoalkane polycarboxylic acids such as diaminopropanoltetraacetic acid [DPTA-OH] and other polyaminoalkanol polycarboxylic acids such as glycol ether diamine tetraacetic acid [GEDTA] and other hydroxyalkyl ether polyamines Polycarboxylic acid etc. are mentioned.

  Examples of the compound having one or more phosphonic acid groups in the molecule include 1 to 4 in a molecule such as alkylaminopoly (alkylphosphonic acid), mono- or polyalkylenepolyaminepoly (alkylphosphonic acid), nitrilopoly (alkylphosphonic acid), and the like. Examples thereof include nitrogen-containing polyphosphonic acids having 6 nitrogen atoms and 1 to 8 phosphonic acid groups, arylphosphonic acids, alkylene polyphosphonic acids, and alkane polyphosphonic acids optionally having a hydroxy group.

  Specific examples of the compound having one or more phosphonic acid groups in the molecule include alkylaminopoly (alkylphosphonic acids) such as ethylaminobis (methylenephosphonic acid) and dodecylaminobis (methylenephosphonic acid), such as ethylenediaminebis. (Methylenephosphonic acid) [EDDPO], ethylenediaminetetrakis (ethylenephosphonic acid), ethylenediaminetetrakis (methylenephosphonic acid) [EDTPO], hexamethylenediaminetetrakis (methylenephosphonic acid), isopropylenediaminebis (methylenephosphonic acid), isopropylenedi Aminetetra (methylenephosphonic acid), propanediaminetetra (ethylenephosphonic acid) [PDTMP], diaminopropanetetra (methylenephosphonic acid) [PDTPO], diethylenetriami Mono, such as penta (ethylene phosphonic acid) [DEPPO], diethylene triamine penta (methylene phosphonic acid) [DETPPO], triethylene tetramine hexa (ethylene phosphonic acid) [TETHP], triethylene tetramine hexa (methylene phosphonic acid) [TTHP] Polyalkylene polyamines Poly (alkylphosphonic acids), for example nitrilotris (methylenephosphonic acid) [NTPO] and other nitrilopoly (alkylphosphonic acids), for example arylphosphonic acids such as phenylphosphonic acid, for example alkylenediphosphonic acids (methylenediphosphonic acid) Etc.), for example, alkanediphosphonic acid (ethylidene diphosphonic acid, 1-hydroxyethylidene-1,1′-diphosphonic acid [HEDPO], which may have a hydroxy group, And alkanepolyphosphonic acids such as 1-hydroxypropylidene-1,1′-diphosphonic acid, 1-hydroxybutylidene-1,1′-diphosphonic acid and the like.

  Examples of N-substituted amino acids include dihydroxyethyl glycine [DHEG] and N-acetyl glycine, and examples of condensed phosphoric acids include tripolyphosphoric acid and hexametaphosphoric acid.

  Among these chelating agents, EDTA, particularly its alkali metal salt, particularly tetrasodium salt (ethylenediaminetetraacetic acid / tetrasodium, EDTA4Na) is preferred.

  A chelating agent may be used independently or may be used in combination of 2 or more types. Further, the amount used may be a concentration that does not easily affect the measurement, and the concentration in the thermostat is generally about 0.00001 to 0.0001 w / w%, preferably 0.00002 to 0.00005 w / w%. In the detergent. The concentration in the detergent may be determined in consideration of how many times it is diluted when added to water in a thermostatic bath, but is usually about 0.001 to 0.1 w / w%, preferably 0.01. It is used alone or in combination of two or more so as to be about 0.05 w / w%.

  In addition, it is preferable that the liquidity of the water in the thermostat at the time of use which added the detergent for thermostats which concerns on this invention is alkalinity in order to prevent the slime of water and generation | occurrence | production of a mold fungus. For example, the pH is preferably 8 to 14, particularly 9 to 12.

  Therefore, it is preferable that a reagent for making the liquid property alkaline, for example, a reagent such as sodium hydroxide or potassium hydroxide, is added to the detergent for the thermostat according to the present invention. At this time, the amount of the alkaline reagent added to the thermostatic bath detergent may be such that the liquidity of the thermostatic bath cleaning agent itself becomes pH 8 to 14, preferably 9 to 12.

  As a temperature chamber cleaning method according to the present invention, the temperature-controlled bath detergent according to the present invention, the temperature of the temperature chamber, preventing the generation of algae and the growth of microorganisms (bacteria etc.), and a concentration that hardly affects the measurement Add so that.

  For example, the concentration of quaternary ammonium salt in a constant temperature bath is generally about 0.001 to 0.1 w / w%, preferably about 0.005 to 0.05 w / w%, and the concentration of the preservative having amphoteric surface activity is the total. About 0.0005 to 0.02 w / w%, preferably about 0.001 to 0.01 w / w%, and the total concentration of surfactants is about 0.005 to 1.0 w / w%, preferably about 0.01 to 0.5 w / w%. The detergent for the thermostatic bath according to the present invention is diluted so that the concentration of the chelating agents contained in the total is about 0.00001 to 0.0001 w / w%, preferably about 0.00002 to 0.00005 w / w%. Use it.

  As the method, for example, the total concentration of quaternary ammonium salts is usually about 1 to 10 w / w%, preferably about 3 to 8 w / w%, and the total concentration of the preservative having amphoteric surface activity is 0.05. ˜10 w / w%, preferably about 1 to 5 w / w%, total concentration of surfactant is about 1 to 20 w / w%, preferably about 5 to 10 w / w%, contained as necessary The detergent for the thermostatic bath of the present invention prepared so that the total concentration of the chelating agent is about 0.001 to 0.1 w / w%, preferably about 0.01 to 0.05 w / w%, What is necessary is just to add and use so that it may become 1000 times dilution.

  Moreover, you may add each reagent so that it may become the above each density | concentration to the water of a thermostat.

In addition, it is preferable that the pH of the water in the prepared thermostat is 8-14 alkalinity, Preferably it is alkaline of pH9-12. A reagent that makes the liquid property alkaline is contained, and when the detergent for the thermostatic bath is added to the water in the thermostatic bath, the liquidity of the water in the thermostatic bath is adjusted to the target pH. do it. In addition, after adding a quaternary ammonium salt or / and an amphoteric surfactant preservative, a surfactant, and a chelating agent contained as necessary to the above-mentioned concentration in the water of the thermostatic bath, the liquidity A reagent that makes the alkalinity alkaline may be added to the water in the thermostatic bath to adjust the liquidity of the water in the thermostatic bath to the desired pH.
The present invention will be described in more detail with reference to the following examples, but the present invention is not limited thereto.

Experimental Example 1 Examination of antibacterial activity (1) Drug and test bacteria 1) Drug Didecyldimethylammonium adipate (DDAA), N-bis (3-aminopropyl) dodecylamine (TAA) and benzalkonium chloride (BC) were used.
2) Test bacteria
Bacillus subtilis ( Bacillus subtilis , NBRC13719), Escherichia coli (E. coli, NBRC3972), Pseudonas aeruginosa (Pseudomonas aeruginosa, NBRC12689), Staphylococcus aureus ( Staphylococcus aureus , NBRC12732), Aspergillus niger (Black mold, NBRC6341), Chaetomium globo NBRC6347), Cladosporium cladosporioides (NBRC6348), Penicillium citrinum (Blue mold genus, NBRC6352) and Trichoderma viride (Nussia mold NBRC31831) were used.

(2) Preparation of medium In a sterile petri dish with a diameter of 90 mm, take an aqueous solution obtained by diluting each drug with distilled water so that the drug concentration when the total volume is adjusted to 20 mL is a predetermined concentration, and then add sterile water to the petri dish. The total amount of sterilized water and aqueous drug solution was 2 mL. Further, 18 mL of a normal agar medium that had been autoclaved and stored at 50 ° C. ± 2 ° C. was placed in a petri dish and solidified to obtain an agar plate medium.
(3) Method for evaluating antibacterial activity Various test bacteria previously cultured in a bouillon medium until 1 × 10 ⁸ cells / mL were diluted 100-fold with sterile physiological saline (1 × 10 ⁶ cells / mL). Next, using the platinum loop, the prepared test bacterial solution was smeared on the surface of the agar plate medium of (2) and cultured at 35 ° C. ± 2 ° C. for 24 hours.
After the culture, the presence or absence of bacterial growth on the medium surface was confirmed, and the minimum drug concentration at which bacterial growth was not observed was defined as the minimum growth inhibitory concentration (MIC, μg / mL) of the drug.

(4) Results Table 1 shows the results.
Moreover, the abbreviation of each microbial cell shown in Table 1 is as follows, respectively.
Bs = Bacillus subtilis = Bacillus subtilis (NBRC13719)
Ec = Escherichia coli = E. coli (NBRC3972)
Pa = Pseudonas aeruginosa = Pseudomonas aeruginosa (NBRC12689)
Sa = Staphylococcus aureus = Staphylococcus aureus (NBRC12732)
An = Aspergillus niger = Black Aspergillus (NBRC6341)
Cg = Chaetomium globosum = Blackfish (NBRC6347)
Cl.cl = Cladosporium cladosporioides (NBRC6348)
Pc = Penicillium citrinum = Blue mold (NBRC6352)
Tv = Trichoderma viride = Aspergillus (NBRC31831)

  As is apparent from the results in Table 1, DDAA and TAA were found to have higher antibacterial activity than BC for all species tested.

Experimental Example 2. Examination of antibacterial activity in the presence of protein (1) Drug and test bacteria 1) Drug Didecyldimethylammonium adipate (DDAA), N-bis (3-aminopropyl) dodecylamine (TAA) and benzalkonium chloride (BC) Was used.
2) Reagent solution • GOT (glutamate oxaloacetate transaminase) solution: L type Wako GOT • G2 substrate enzyme solution of G2 kit (manufactured by Wako Pure Chemical Industries, Ltd.) and GOT • L2α-KG solution with distilled water It prepared so that it might become / w% aqueous solution.
・ GPT (glutamate pyruvate transaminase) solution: L type Wako GPT ・ J2 kit (manufactured by Wako Pure Chemical Industries, Ltd.) GPT substrate enzyme solution and GPT • L2α-KG solution in distilled water with 0.2 w / w% aqueous solution It prepared so that it might become.
Serum: Control Serum II Wako (manufactured by Wako Pure Chemical Industries, Ltd.) was prepared in distilled water to make a 0.2 w / w% aqueous solution.
3) Test bacteria
Bacillus subtilis ( Bacillus subtilis , NBRC13719), Escherichia coli (E. coli, NBRC3972), Pseudonas aeruginosa (Pseudomonas aeruginosa, NBRC12689) and Staphylococcus aureus ( Staphylococcus aureus , NBRC12732) were used.

(2) Preparation of medium In a sterile petri dish with a diameter of 90 mm, an aqueous solution obtained by diluting each drug with distilled water is adjusted so that the drug concentration when the total volume is adjusted to 20 mL is the prescribed concentration, and each component in the reagent (GOT , GPT, serum) Take the aqueous drug solution and the reagent solution in a petri dish so that the final concentration is 100 μg / mL when the total volume is adjusted to 20 mL, then sterilize water in the petri dish, The total amount with the reagent solution was 2 mL. Further, 18 mL of a normal agar medium that had been autoclaved and stored at 50 ° C. ± 2 ° C. was placed in a petri dish and solidified to obtain an agar plate medium. As a control, an agar plate medium containing pure water instead of the drug solution was also prepared.
(3) Method for evaluating antibacterial activity Various test bacteria previously cultured in a bouillon medium until 1 × 10 ⁸ cells / mL were diluted 100-fold with sterile physiological saline (1 × 10 ⁶ cells / mL). Next, using the platinum loop, the prepared test bacterial solution was smeared on the surface of the agar plate medium of (2) and cultured at 35 ° C. ± 2 ° C. for 24 hours.
After the culture, the presence or absence of bacterial growth on the medium surface was confirmed, and the minimum drug concentration at which bacterial growth was not observed was defined as the minimum growth inhibitory concentration (MIC, μg / mL) of the drug.

(4) Results Table 2 shows the results.
Moreover, the abbreviation of each microbial cell shown in Table 2 is as follows, respectively.
Bs = Bacillus subtilis = Bacillus subtilis (NBRC13719)
Ec = Escherichia coli = E. coli (NBRC3972)
Pa = Pseudonas aeruginosa = Pseudomonas aeruginosa (NBRC12689)
Sa = Staphylococcus aureus = Staphylococcus aureus (NBRC12732)

As apparent from Table 2, DDAA and TAA did not change the antibacterial activity against various bacteria even under conditions where proteins such as enzymes and serum were present, and were found to be preferable for use as a detergent for a thermostatic bath.
On the other hand, it was found that BC is not preferable for use in a thermostatic bath detergent because its antibacterial activity decreases when proteins coexist.

Experimental Example 3. Examination of bactericidal power (1) Drug and test bacteria 1) Drug Didecyldimethylammonium adipate (DDAA), N-bis (3-aminopropyl) dodecylamine (TAA) and benzalkonium chloride (BC) are distilled respectively. It prepared so that it might become 1 w / w% with water.
2) Reagent solution / GOT solution: L-type Wako GOT / J2 kit (manufactured by Wako Pure Chemical Industries, Ltd.) GOT substrate enzyme solution and GOT / L2α-KG solution are made into 0.1 w / w% aqueous solution with distilled water. It was prepared as follows.
· GPT solution: L type Wako GPT substrate enzyme solution and GPT · L2α-KG solution GPT · J2 kit (manufactured by Wako Pure Chemical Industries, Ltd.) was prepared so as to 0.1 w / w% aqueous solution with distilled water .
Serum: Control serum II Wako (manufactured by Wako Pure Chemical Industries, Ltd.) was prepared with distilled water so as to be a 0.1 w / w% aqueous solution.
3) Test bacteria
Bacillus subtilis ( Bacillus subtilis , NBRC13719), Escherichia coli (E. coli, NBRC3972), Pseudonas aeruginosa (Pseudomonas aeruginosa, NBRC12689) and Staphylococcus aureus ( Staphylococcus aureus , NBRC12732) were used.

(2) Bactericidal power evaluation method
i) Various test bacteria previously cultured in a bouillon medium until 1 × 10 ⁸ cells / mL were diluted 100-fold with sterile physiological saline to obtain a test cell solution (1 × 10 ⁶ cells / mL).
ii) In a 20 mL glass test tube sterilized by dry heat in advance, when the total amount of the drug aqueous solution, reagent solution, and test bacterial solution prepared in i) above is adjusted to 10 mL, the final concentration of the drug is 50 μg. The final concentration of the reagent was 100 μg / mL and the test bacteria was 1 × 10 ⁵ cells / mL, and the mixture was sufficiently stirred.
iii) After a lapse of a certain time (5 minutes, 10 minutes, 15 minutes), the solution obtained in ii) above was smeared on the surface of a normal agar plate medium prepared in advance using a platinum loop, and 35 ° C. The cells were cultured at ± 2 ° C for 24 hours.
iv) After culturing, the presence or absence of bacterial growth on the surface of the medium was confirmed.

(3) Results Table 3 shows the results.
In Table 3, the case where the growth of bacteria was observed on the surface of the medium was indicated by +, and the case where the growth of bacteria was not observed on the surface of the medium was indicated by-.
The abbreviations of the cells shown in Table 3 are as described in Experimental Example 2.

  As is clear from Table 3, DDAA and TAA are more susceptible to nosocomial infections such as Pseudomonas aeruginosa (Pa) and Staphylococcus aureus (Sa), even under the presence of proteins such as enzymes and serum, compared to BC. It was found that bactericidal power against the bacteria involved is high. In addition, although data are not shown, TAA, a preservative having amphoteric surfactant activity, shows high bactericidal activity against Pseudomonas aeruginosa and Staphylococcus aureus even at a low concentration of 10 μg / mL, and the effect is It did not change even in the presence of proteins such as enzymes and serum.

Example 1. Examination of the bactericidal power of the detergent for the temperature-controlled bath of the present invention From the results of Experimental Examples 1 to 3, didecyldimethylammonium adipate and N-bis (3-aminopropyl) dodecylamine are excellent in both antibacterial power and bactericidal power. Thus, the detergent for a thermostatic bath of the present invention containing this was prepared. And the bactericidal power with respect to various microbes was compared about the detergent for thermostats of this invention, and the conventional detergent for thermostats.

(1) Preparation of detergent for constant temperature bath of the present invention Didecyldimethylammonium adipate 5.6 w / w%, N-bis (3-aminopropyl) dodecylamine 2.4 w / w%, EDTA4Na 0.02 w / w%, polyoxy Ethylene alkyl ether was dissolved in pure water so as to be 4.5 w / w% and sodium lauroyl sarcosine 0.5 w / w%, and adjusted to pH 11.4 with an aqueous sodium hydroxide solution, which was used as a thermostatic bath detergent of the present invention.
(2) Preparation of a conventional detergent for a thermostatic bath A mixture of polyoxyethylene nonylphenyl ether: monoethenolamine: distilled water = 20: 5: 5: 70 is prepared, It was set as the detergent for thermostats.
(3) Test bacteria
Bacillus subtilis (B. subtilis, NBRC13719), Escherichia coli (E. coli, NBRC3972), Pseudonas aeruginosa (Pseudomonas aeruginosa, NBRC12689), Staphylococcus aureus ( Staphylococcus aureus , NBRC12732) were used.

(4) Bactericidal power evaluation method
i) Various test bacteria previously cultured in a bouillon medium until 1 × 10 ⁸ cells / mL were diluted 100-fold with sterile physiological saline to obtain a test cell solution (1 × 10 ⁶ cells / mL).
ii) In a 20 mL glass test tube sterilized by dry heat in advance, the concentration when the total temperature of the detergent for the thermostatic bath of the present invention or the conventional thermostatic bath cleaner is adjusted to 10 mL is a predetermined concentration (0.05 w / w%, 0.1w / w%, 0.2w / w%). Next, the test bacterium solution obtained in i) above was put in the test tube so that the total of the detergent for the thermostat and the test bacterium solution was 10 μl.
iii) After a lapse of a certain time (5 minutes, 10 minutes, 15 minutes), smear the test bacterial solution obtained in ii) above on the surface of a normal agar plate medium prepared in advance using a platinum loop, The cells were cultured at 35 ° C. ± 2 ° C. for 24 hours.
iv) After the culture, the presence or absence of growth of bacteria on the surface of the medium was confirmed, and the minimum concentration of the detergent for constant temperature bath in which the growth of bacteria was not observed was determined.

(5) Results Table 4 shows the results.
In Table 4, the case where the growth of bacteria was observed on the surface of the medium was shown as +, and the case where the growth of bacteria was not observed on the surface of the medium was shown as-.
The abbreviations of the cells shown in Table 4 are as described in Experimental Example 2.
In Table 4, the final concentration of each component in each concentration of the detergent for the thermostatic bath of the present invention is as follows.

When the concentration of the detergent for the temperature chamber is 0.05 w / w%, didecyldimethylammonium adipate is 0.0028 w / w%, N-bis (3-aminopropyl) dodecylamine is 0.0012 w / w%, EDTA4Na is 0.00001 w / w%, polyoxyethylene alkyl ether is 0.00225 w / w%, and lauroyl sarcosine sodium is 0.00025 w / w%.
When the concentration of the detergent for the temperature chamber is 0.1 w / w%, didecyldimethylammonium adipate is 0.0056 w / w%, N-bis (3-aminopropyl) dodecylamine is 0.0024 w / w%, EDTA4Na is 0.00002 w / w%, polyoxyethylene alkyl ether is 0.0045 w / w%, and lauroyl sarcosine sodium is 0.0005 w / w%.
When the concentration of the detergent for the temperature chamber is 0.2 w / w%, didecyldimethylammonium adipate is 0.0112 w / w%, N-bis (3-aminopropyl) dodecylamine is 0.0048 w / w%, EDTA4Na is 0.00004 w / w%, polyoxyethylene alkyl ether is 0.009 w / w%, and lauroyl sarcosine sodium is 0.001 w / w%.

  As is apparent from Table 4, the temperature-controlled bath detergent of the present invention has a higher bactericidal power than conventional temperature-controlled bath cleaners, and particularly has a bactericidal power against Pseudomonas aeruginosa (Pa) and Staphylococcus aureus (Sa). From the fact that it is expensive, it was found that it can sufficiently meet the demand for prevention and prevention of nosocomial infections.

Example 2 Time stability test 1
(1) Preparation of detergent for constant temperature bath of the present invention Didecyldimethylammonium adipate 5.6 w / w%, EDTA4Na 0.02 w / w%, polyoxyethylene alkyl ether 4.5 w / w%, lauroyl sarcosine sodium 0.5 w / w% Then, it was dissolved in pure water, adjusted to pH 11.4 with an aqueous sodium hydroxide solution, and used as the thermostatic bath detergent of the present invention.

(2) Stability test with time The stock solution of the detergent of the present invention prepared in (1) above or a solution diluted with pure water to 0.2 w / w% is placed in a constant temperature water bath at room temperature or 40 ° C. After covering with a transparent film, it was allowed to stand. After standing for a predetermined period, the appearance, specific gravity (20 ° C.), viscosity (mm ² / s, 25 ° C.) , pH, and absorbance at 800 to 400 nm, 340 nm, and 300 nm were measured. Further, the antibacterial activity was tested in the same manner as in Experimental Example 1, and the bactericidal activity in the treatment for 5 minutes was tested in the same manner as in Example 1. Table 5 shows the results when stored at room temperature, and Table 6 shows the results when stored at 40 ° C.

In addition, the concentration of each chemical in the detergent for a thermostatic bath diluted to 0.2 w / w% is 0.0112 w / w% for didecyldimethylammonium adipate, 0.00004 w / w% for EDTA4Na, and 0.009 for polyoxyethylene alkyl ether. w / w%, lauroyl sarcosine sodium is 0.001w / w%.
Further, the abbreviations of the bacterial cells shown in each table are as described in Experimental Example 2.

As is apparent from Tables 5 and 6, the constant temperature bath detergent containing didecyldimethylammonium adipate (a quaternary ammonium salt according to the present invention) is 0.2 w / w% of the use mode even in the stock solution. Even when the solution was stored for a long period of 6 months at room temperature, there was almost no change in specific gravity and viscosity. Moreover, the antibacterial power and the bactericidal power maintained the effect immediately after preparation. Furthermore, the detergent for the thermostatic bath of the present invention containing didecyldimethylammonium adipate has almost no practical change in specific gravity and viscosity even under the condition of standing at 40 ° C. for 6 months. In addition, the antibacterial and bactericidal effects were maintained immediately after preparation.
From the above, it was found that the thermostatic bath detergent of the present invention is excellent in long-term storage stability.

Example 3 Aging stability test 2
(1) Preparation of the thermostatic bath detergent of the present invention
N-bis (3-aminopropyl) dodecylamine 2.4 w / w%, EDTA4Na 0.02 w / w%, polyoxyethylene alkyl ether 4.5 w / w%, lauroyl sarcosine sodium 0.5 w / w% It was dissolved and adjusted to pH 11.4 with an aqueous sodium hydroxide solution, and used as the thermostatic bath detergent of the present invention.

(2) Stability test with time The stock solution of the detergent of the present invention prepared in (1) above or a solution diluted with pure water to 0.2 w / w% is placed in a constant temperature water bath at room temperature or 40 ° C. After covering with a transparent film, it was allowed to stand. After standing for a predetermined period, the appearance, specific gravity (20 ° C.), viscosity (mm ² / s, 25 ° C.) , pH, and absorbance at 800 to 400 nm, 340 nm, and 300 nm were measured. Further, the antibacterial activity was tested in the same manner as in Experimental Example 1, and the bactericidal activity in the treatment for 5 minutes was tested in the same manner as in Example 1. Table 7 shows the results when stored at room temperature, and Table 8 shows the results when stored at 40 ° C.

The concentration of each drug in the thermostatic bath detergent diluted to 0.2 w / w% is 0.0048 w / w% for N-bis (3-aminopropyl) dodecylamine, 0.00004 w / w% for EDTA4Na, Oxyethylene alkyl ether is 0.009 w / w% and lauroyl sarcosine sodium is 0.001 w / w%.
Further, the abbreviations of the bacterial cells shown in each table are as described in Experimental Example 2.

As is clear from Tables 7 and 8, the detergent for the thermostatic bath of the present invention containing N-bis (3-aminopropyl) dodecylamine (a preservative having amphoteric surface activity according to the present invention) is a stock solution. Even when the 0.2 w / w% solution of the usage mode was stored for a long period of 6 months at room temperature, there was almost no change in specific gravity, viscosity and pH. Moreover, the antibacterial power and the bactericidal power maintained the effect immediately after preparation. Furthermore, the thermostatic bath detergent of the present invention shows almost no changes that cause practical problems with respect to specific gravity, viscosity, and pH even under the condition of being allowed to stand at 40 ° C. for 6 months. In addition, both the bactericidal power and the effect immediately after the preparation were maintained.
From the above, it was found that the thermostatic bath detergent of the present invention is excellent in long-term storage stability.

  Further, as is clear from comparison with the results of Table 2 (Tables 5 and 6), the thermostatic bath detergent of the present invention containing N-bis (3-aminopropyl) dodecylamine has a pH even under long-term storage. There is almost no fluctuation. It can also be seen that the antibacterial activity against Pseudomonas aeruginosa is also superior.

Example 4 Aging stability test 3
The stock solution of the detergent of the present invention prepared in Example 1 or a solution diluted with pure water to 0.2 w / w% was placed in a constant temperature water bath at room temperature or 40 ° C., and the upper part was covered with a transparent film, and then left standing. did. After standing for a predetermined period, the appearance, specific gravity (20 ° C.), viscosity (mm ² / s, 25 ° C.) , pH, and absorbance at 800 to 400 nm, 340 nm, and 300 nm were measured. Further, the antibacterial activity was tested in the same manner as in Experimental Example 1, and the bactericidal activity in the treatment for 5 minutes was tested in the same manner as in Example 1. Table 9 shows the results when stored at room temperature, and Table 10 shows the results when stored at 40 ° C.

  The concentration of each chemical in the thermostatic bath detergent diluted to 0.2 w / w% is 0.0112 w / w% for didecyldimethylammonium adipate, 0.0048 w / w for N-bis (3-aminopropyl) dodecylamine. w%, EDTA4Na is 0.00004 w / w%, polyoxyethylene alkyl ether is 0.009 w / w%, and lauroyl sarcosine sodium is 0.001 w / w%.

In Tables 9 and 10, “-” indicates a case where measurement was not performed.
Further, the abbreviations of the bacterial cells shown in each table are as described in Experimental Example 2.

As is apparent from Tables 9 and 10, the constant temperature bath detergent of the present invention can be used as a stock solution, a 0.2 w / w% solution of the usage mode, or a long-term storage of 6 months at room temperature. Little change was seen in viscosity, pH and absorbance. Moreover, the antibacterial power and the bactericidal power maintained the effect immediately after preparation. Furthermore, the detergent for the thermostatic bath of the present invention shows almost no change that causes practical problems with respect to appearance, specific gravity, viscosity, pH, and absorbance even under the condition of being left at 40 ° C. for 6 months. Moreover, the antibacterial power and the bactericidal power maintained the effect immediately after preparation.
From the above, it was found that the thermostatic bath detergent of the present invention is excellent in long-term storage stability.

Further, as is apparent from comparison with the results of Example 2 (Tables 5 and 6) and the results of Example 3 (Tables 7 and 8), the quaternary ammonium salt according to the present invention and an antiseptic having amphoteric surface activity. The detergent for the thermostatic bath of the present invention containing both of the agents has almost no change in appearance, specific gravity, pH, and absorbance even after long-term storage, and E. coli (Ec) and Pseudomonas aeruginosa (Pa) even after long-term storage. Since the high antibacterial power can be maintained, it can be seen that it is further superior to the detergent for a thermostatic bath of the present invention containing each of them alone.

The present invention prevents the generation of microorganisms (bacteria etc.) generated in the water in the thermostat bath and the accompanying decrease in measurement accuracy in scientific instruments having a thermostat bath using water as a medium, particularly in an automatic analyzer. In addition, it prevents the generation and adhesion of bubbles on the outer wall of the reaction vessel in the thermostat, and it itself absorbs at the measurement wavelength (300 to 900 nm) due to decomposition of some of its constituent components when storing the stock solution This is to provide a thermostatic bath cleaner that has a higher sterilizing effect than the conventional one. The present invention provides an excellent temperature-controlled bath detergent that can take advantage of the advantages of quickness, high efficiency, high accuracy, and ease of operation.

Claims (14)

(1) A quaternary ammonium salt represented by the following general formula [I]

(Wherein R _{1 to} R ₄ each independently represents an alkyl group, provided that at least one of R _{1 to} R ₄ is an alkyl group having 8 to 18 carbon atoms, and at least one is carbon. the number is 1 to 3 lower alkyl groups .X ^-. represents an anion derived from a carboxylic acid having 2 or more carbon atoms) or / and
(2) N-bis (3-aminopropyl) dodecylamine, 3-aminopropyl-dodecylpropanediamine, 1,3-propanediamine-N-3-aminopropyl-N-dodecyl, alkylaminoglycine, alkyldiaminoethylglycine A preservative having an amphoteric surfactant ability selected from hydrochloride, alkylaminoethylaminoethylglycine, alkylaminopropylaminoacetic acid, and alkylpolyaminoethylglycine ;
(3) Surfactant (excluding those included in (1) and (2) above)
Detergent for constant temperature bath. The detergent according to claim 1, wherein the quaternary ammonium salt represented by the general formula [I] is dialkyldimethylammonium adipate or dialkyldimethylammonium propionate. The detergent according to claim 1, wherein the quaternary ammonium salt is didecyldimethylammonium adipate. The detergent according to any one of claims 1 to 3, wherein the preservative having amphoteric surface active ability is N-bis (3-aminopropyl) dodecylamine. The detergent according to any one of claims 1 to 4 , wherein the surfactant is a nonionic surfactant or an anionic surfactant. The detergent according to any one of claims 1 to 5 , wherein the surfactant is polyoxyethylene alkyl ether or / and lauroyl sarcosine sodium. (1) A quaternary ammonium salt represented by the general formula [I], and (2) N-bis (3-aminopropyl) dodecylamine, 3-aminopropyl-dodecylpropanediamine, 1,3-propanediamine-N Preservative with amphoteric surfactant ability selected from -3-aminopropyl-N-dodecyl, alkylaminoglycine, alkyldiaminoethylglycine hydrochloride, alkylaminoethylaminoethylglycine, alkylaminopropylaminoacetic acid, and alkylpolyaminoethylglycine The detergent for thermostats of Claim 1 containing an agent and (3) surfactant (however, except what is contained in said (1) and (2)). The quaternary ammonium salt represented by the general formula [I] is didecyldimethylammonium adipate, the preservative having amphoteric surface activity is N-bis (3-aminopropyl) dodecylamine, and the surfactant is polyoxyethylene. The detergent according to claim 7, which is an alkyl ether or / and lauroyl sarcosine sodium. Furthermore, the detergent in any one of Claims 1-8 containing a chelating agent. The detergent according to claim 9 , wherein the chelating agent is ethylenediaminetetraacetate. Furthermore, the detergent in any one of Claims 1-10 containing the reagent which makes liquidity alkaline. The detergent according to claim 11 , wherein the reagent that makes the liquid alkaline is sodium hydroxide. The detergent according to any one of claims 1 to 12 , wherein the liquid is alkaline having a pH of 10 to 12 . A cleaning method for a thermostatic bath, wherein the detergent according to claim 1 is added to the thermostatic bath.