Organic ammonium salt and hydrogen-bonding material treating agent using the same

Abstract

To provide a solid composition useful for various treatments of a hydrogen bonding material such as a biological sample.SOLUTION: A compound of formula (I) Wherein each R independently represents a linear or branched hydroxyalkyl group having one or more hydroxyl groups and an alkyl moiety having 1 to 10 carbon atoms; N represents an integer of 1 to 3. The solid composition contains the organic ammonium salt and a biological sample, wherein the mass ratio of the organic ammonium salt to the biological sample is 10:1 to 0.1:1.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to an organic ammonium salt and a treatment agent for hydrogen-bonding materials using the same. [Background technology]

[0002] Many biological samples, such as biocatalysts (e.g., enzymes and yeast), peptides, proteins, nucleic acids, and poorly soluble polysaccharides (e.g., cellulose), are susceptible to the influence of temperature, pH, solvents, or intermolecular electrostatic repulsion, resulting in a loss of their three-dimensional molecular structure and a decrease in their activity, i.e., catalytic ability. Therefore, when preserving biocatalytic activity, using them in biocatalytic reactions, or storing biological samples, it is necessary to maintain the three-dimensional structure of the active site and the three-dimensional structure of the amino acid residues.

[0003] Known methods for long-term storage of biocatalysts and biological samples include freeze-drying in a powder state and cryopreservation, in which the sample is dissolved in a solution at a low concentration and at an extremely low temperature.

[0004] Generally, storage in solution is desirable for ease of operation; however, cryopreservation requires special equipment, and there are problems with the three-dimensional structure of the biocatalyst being destroyed by the ice generated during freezing, and with the structure of the biocatalyst changing and its activity decreasing when the frozen solution is thawed for use. Furthermore, the storage concentration is generally low, around 1 to 3 mg / mL, for example, in the case of urease and catalase, making efficient storage difficult.

[0005] Attempts have been made to add stabilizers to prevent the biocatalyst from being deactivated and maintain the enzyme activity. For example, a method of preserving uricase using polyhydric alcohols such as glycerin and sorbitol (Patent Document 1) and a method of stabilizing cholesterol oxidase by adding bovine serum albumin and sugars to a solution containing cholesterol oxidase (Patent Document 2) have been proposed. However, these methods have the problem of a decrease in enzyme activity depending on the storage concentration, storage temperature, and storage period.

[0006] To solve the above problems, the applicant discovered that ionic liquids that are liquid at room temperature can be used to preserve biological catalysts and biological samples (Patent Documents 3 to 5). Although ionic liquids that are liquid at room temperature have excellent affinity for biological samples and are easy to preserve, they are not suitable for applications that require a solid state at room temperature, such as clinical diagnostic reagents and biosensors, and further improvements were necessary. Furthermore, when storing ionic liquids in a liquid state, it is difficult to preserve biological samples at high concentrations due to the solubility of the biological sample and the liquid. For this reason, freeze-drying and frozen storage have been used to achieve a solid state, but these methods have the problem of providing little long-term stability for biological samples.

[0007] As an example of a solid enzyme stabilizer, a case has been disclosed in which a polypeptide derived from a plant, such as a hydrolysate of soybean, is used (Patent Document 6), but the stabilizing effect was insufficient.

[0008] In addition to such biological sample preservation materials, there is a demand for processing agents that enable various types of processing of hydrogen-bonding materials such as biological samples, for example, processing agents that have excellent solubility or dispersibility when water, solvents, etc. are added after the hydrogen-bonding material is made into a solid composition, and novel compounds that have good affinity with hydrogen-bonding materials and make this possible. [Prior art documents] [Patent documents]

[0009] [Patent Document 1] Japanese Patent Application Publication No. 06-70798 [Patent Document 2] Japanese Patent Application Publication No. 08-187095 [Patent Document 3] Japanese Patent Application Laid-Open No. 2014-131974 [Patent Document 4] Japanese Patent Application Laid-Open No. 2014-131975 [Patent Document 5] International Publication No. 2015 / 156398 [Patent Document 6] Japanese Patent Application Laid-Open No. 2006-42757 Summary of the Invention [Problem to be solved by the invention]

[0010] The present invention has been made in view of the above circumstances, and one of its objectives is to provide a novel compound that is useful for various treatments of hydrogen-bonding materials.

[0011] Another object of the present invention is to provide a treatment agent that is useful for various treatments of hydrogen-bonding materials.

[0012] Another object of the present invention is to provide a processing agent that is excellent at maintaining the three-dimensional structure of a biological sample in a solid state, even at high concentrations, high temperatures, and for long periods of time, and a biological sample solution that is excellent at maintaining the three-dimensional structure of a biological sample in a solution state, such as an aqueous solution, even at high concentrations, high temperatures, and for long periods of time.

[0013] Another object of the present invention is to provide a treatment agent that has excellent solubility and dispersibility for organic and inorganic hydrogen-bonding materials, and that has excellent solubility or dispersibility when the resulting solution or dispersion is made into a solid composition and then water, a solvent, or the like is added thereto. [Means for solving the problem]

[0014] In order to solve the above problems, the organic ammonium salt of the present invention is represented by the following formula (I):

[0015] [ka] (wherein R each independently represent a hydroxyalkyl group having one or more hydroxyl groups and a linear or branched alkyl moiety of 1 to 10 carbon atoms, which may contain an oxygen atom; a carboxyalkyl group having one or more carboxy groups and a linear or branched alkyl moiety of 1 to 10 carbon atoms, which may contain an oxygen atom; or a hydroxycarboxyalkyl group having one or more hydroxyl groups and one or more carboxy groups, which linear or branched alkyl moiety of 1 to 10 carbon atoms, which may contain an oxygen atom; or a hydroxycarboxyalkyl group having one or more hydroxyl groups and one or more carboxy groups, which linear or branched alkyl moiety of 1 to 10 carbon atoms, which may contain an oxygen atom; and n represents an integer of 0 to 4.) The hydrogen-bonding material treating agent of the present invention contains the organic ammonium salt. In a preferred example of the hydrogen-bonding material treating agent, the hydrogen-bonding material is a biological sample (hereinafter, in this case, also referred to as a biological sample treating agent). The solid composition of the present invention contains the hydrogen-bonding material treating agent and the hydrogen-bonding material. The biological sample solution of the present invention contains the hydrogen-bonding material treating agent (biological sample treating agent), a biological sample, and a solvent. [Effects of the Invention]

[0016] According to the present invention, a novel compound is provided which is useful for various treatments of hydrogen-bonding materials. Also provided is a treatment agent useful for treating various hydrogen-bonding materials such as biological samples.

[0017] According to the present invention, a processing agent is provided that is excellent at maintaining the three-dimensional structure of a biological sample in a solid state, even at high concentrations, high temperatures, and for long periods of time, as well as a biological sample solution that is excellent at maintaining the three-dimensional structure of a biological sample in a solution state, such as an aqueous solution, even at high concentrations, high temperatures, and for long periods of time.

[0018] According to the present invention, there is provided a treatment agent which has excellent solubility and dispersibility in organic and inorganic hydrogen-bonding materials, and which has excellent solubility or dispersibility when the resulting solution or dispersion is made into a solid composition and then water, a solvent, etc. is added. DETAILED DESCRIPTION OF THE INVENTION

[0019] The present invention will be described in detail below. (organic ammonium salts)

[0020] In the cation of the organic ammonium salt of the present invention, R in formula (I) represents a hydroxyalkyl group, a carboxyalkyl group, or a hydroxycarboxyalkyl group.

[0021] The hydroxyalkyl group has one or more hydroxyl groups, and the alkyl moiety is linear or branched and preferably has 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms, and the alkyl moiety may contain an oxygen atom.

[0022] The carboxyalkyl group has one or more carboxy groups, and the alkyl moiety is linear or branched and preferably has 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms, and the alkyl moiety may contain an oxygen atom.

[0023] The hydroxycarboxyalkyl group has one or more hydroxyl groups and one or more carboxy groups, and the alkyl moiety is linear or branched and preferably has 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms, and the alkyl moiety may contain an oxygen atom.

[0024] When the alkyl moiety contains an oxygen atom, the oxygen atom forms, for example, an ether bond, a carbonyl group, an ester bond, an amide bond, a urea bond, or a urethane bond in the alkyl moiety. Therefore, in the present invention, the phrase "the alkyl moiety contains an oxygen atom" includes cases where the alkyl moiety is interrupted by a group that also contains a heteroatom such as a nitrogen atom as an atomic group containing an oxygen atom.

[0025] Examples of the hydroxyalkyl group include monohydroxyalkyl groups and polyhydroxyalkyl groups, and specific examples thereof include hydroxyalkyl groups, hydroxyalkoxyalkyl groups, alkoxyhydroxyalkyl groups, and hydroxypolyalkyleneoxyalkyl groups in which the alkyl moiety does not contain an oxygen atom.

[0026] The monohydroxyalkyl group is not particularly limited, and examples thereof include a hydroxymethyl group, a 1-hydroxyethyl group, a 2-hydroxyethyl group, a 1-hydroxypropan-1-yl group, a 2-hydroxypropan-1-yl group, a 3-hydroxypropan-1-yl group, a 1-hydroxypropan-2-yl group, a 2-hydroxypropan-2-yl group, a 1-hydroxybutan-1-yl group, a 2-hydroxybutan-1-yl group, a 3-hydroxybutan-1-yl group, a 4-hydroxybutan-1-yl group, a 1-hydroxy-2-methylpropan-1-yl group, a 2-hydroxypropan-2-yl group, a 1-hydroxybutan-1-yl group, a 2-hydroxybutan-1-yl group, a 4-hydroxybutan-1-yl group, a 1-hydroxy-2-methylpropan-1-yl group, a 2-hydroxybut ... Examples of the monohydroxyalkyl group include hydroxy-2-methylpropan-1-yl group, 3-hydroxy-2-methylpropan-1-yl group, 1-hydroxybutan-2-yl group, 2-hydroxybutan-2-yl group, 3-hydroxybutan-2-yl group, 4-hydroxybutan-2-yl group, 1-hydroxy-2-methylpropan-2-yl group, 5-hydroxypentan-1-yl group, 6-hydroxyhexan-1-yl group, 7-hydroxyheptan-1-yl group, 8-hydroxyoctan-1-yl group, 9-hydroxynonan-1-yl group, and 10-hydroxydecan-1-yl group. The monohydroxyalkyl group preferably has 1 to 10 carbon atoms, more preferably has 1 to 6 carbon atoms, and even more preferably has 1 to 3 carbon atoms.

[0027] The polyhydroxyalkyl group is not particularly limited, but examples thereof include di-, tri-, tetra-, penta-, hexa-, hepta-, or octahydroxyalkyl groups. Specific examples thereof include dihydroxyethyl groups such as 1,2-dihydroxyethyl groups; dihydroxypropan-1-yl groups such as 1,2-dihydroxypropan-1-yl groups and 2,3-dihydroxypropan-1-yl groups; dihydroxypropan-2-yl groups such as 1,2-dihydroxypropan-2-yl groups and 1,3-dihydroxypropan-2-yl groups; trihydroxypropan-1-yl groups; trihydroxypropan-2-yl groups; 1,2-dihydroxybutan-1-yl groups; 1,3-dihydroxybutan-1-yl groups; Dihydroxybutan-1-yl groups such as dihydroxybutan-1-yl group, 1,4-dihydroxybutan-1-yl group, 2,3-dihydroxybutan-1-yl group, 2,4-dihydroxybutan-1-yl group, and 3,4-dihydroxybutan-1-yl group; trihydroxybutan-1-yl groups such as 1,2,3-trihydroxybutan-1-yl group, 1,2,4-trihydroxybutan-1-yl group, 1,3,4-trihydroxybutan-1-yl group, and 2,3,4-trihydroxybutan-1-yl group; group; tetrahydroxybutan-1-yl group; dihydroxy-2-methylpropan-1-yl groups such as 1,2-dihydroxy-2-methylpropan-1-yl group, 1,3-dihydroxy-2-methylpropan-1-yl group, and 2,3-dihydroxy-2-methylpropan-1-yl group; trihydroxy-2-methylpropan-1-yl group; tetrahydroxy-2-methylpropan-1-yl group; 1,2-dihydroxybutan-2-yl group, 1,3-dihydroxybutan-2-yl group, 1,4 -Dihydroxybutan-2-yl groups such as a dihydroxybutan-2-yl group, a 2,3-dihydroxybutan-2-yl group, a 2,4-dihydroxybutan-2-yl group, or a 3,4-dihydroxybutan-2-yl group; trihydroxybutan-2-yl groups such as a 1,2,3-trihydroxybutan-2-yl group, a 1,2,4-trihydroxybutan-2-yl group, a 1,3,4-trihydroxybutan-2-yl group, or a 2,3,4-trihydroxybutan-2-yl group; tetrahydroxybutan-2-yl group;Examples of suitable polyhydroxyalkyl groups include 1,3-dihydroxy-2-methylpropan-2-yl groups, 1,3-dihydroxy-2-ethylpropan-2-yl groups, 1,3-dihydroxy-2-hydroxymethylpropan-2-yl groups, di-, tri-, tetra-, or pentahydroxypentan-1-yl groups, di-, tri-, tetra-, penta-, or hexahydroxyhexane-1-yl groups, di-, tri-, tetra-, penta-, hexa-, or heptahydroxyheptan-1-yl groups, and di-, tri-, tetra-, penta-, hexa-, hepta-, or octahydroxyoctan-1-yl groups. Polyhydroxyalkyl groups preferably have 2 to 6 hydroxyl groups and 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms. Preferred examples of suitable polyhydroxyalkyl groups include branched polyhydroxyalkyl groups represented by the following formula (II):

[0028] [ka] (In the formula, R 11 represents a hydrogen atom, a linear alkyl group having 1 to 4 carbon atoms, or a linear monohydroxyalkyl group having 1 to 4 carbon atoms.

[0029] Of the above polyhydroxyalkyl groups, 2,3-dihydroxypropan-1-yl, 1,3-dihydroxypropan-2-yl, 1,3-dihydroxy-2-ethylpropan-2-yl, 1,3-dihydroxy-2-hydroxymethylpropan-2-yl, and pentahydroxyhexan-1-yl groups are preferred.

[0030] Examples of the carboxyalkyl group include monocarboxyalkyl groups and polycarboxyalkyl groups, and specific examples thereof include those in which the hydroxyl groups of the mono-, di-, tri-, tetra-, penta-, hexa-, hepta-, or octahydroxyalkyl groups exemplified above are substituted with carboxy groups.

[0031] The monocarboxyalkyl group is not particularly limited, and examples thereof include a carboxymethyl group, a 1-carboxyethyl group, a 2-carboxyethyl group, a 1-carboxypropan-1-yl group, a 2-carboxypropan-1-yl group, a 3-carboxypropan-1-yl group, a 1-carboxypropan-2-yl group, a 2-carboxypropan-2-yl group, a 1-carboxybutan-1-yl group, a 2-carboxybutan-1-yl group, a 3-carboxybutan-1-yl group, a 4-carboxybutan-1-yl group, a 1-carboxy-2-methylpropan-1 ... Examples of the carboxyalkyl group include carboxy-2-methylpropan-1-yl group, 3-carboxy-2-methylpropan-1-yl group, 1-carboxybutan-2-yl group, 2-carboxybutan-2-yl group, 3-carboxybutan-2-yl group, 4-carboxybutan-2-yl group, 1-carboxy-2-methylpropan-2-yl group, 5-carboxypentan-1-yl group, 6-carboxyhexane-1-yl group, 7-carboxyheptan-1-yl group, 8-carboxyoctan-1-yl group, 9-carboxynonan-1-yl group, and 10-carboxydecan-1-yl group. The carboxyalkyl group preferably has 1 to 10 carbon atoms, and more preferably has 1 to 6 carbon atoms.

[0032] The hydroxycarboxyalkyl group is not particularly limited, but examples thereof include those in which some of the hydroxyl groups of the di-, tri-, tetra-, penta-, hexa-, hepta-, or octahydroxyalkyl groups exemplified above have been substituted with carboxy groups.

[0033] In the organic ammonium salt of the present invention, n in formula (I) is an integer of 0 to 4, preferably an integer of 1 to 4. When R in formula (I) is a polyhydroxyalkyl group, n is preferably 1 to 4, more preferably 1 to 3, still more preferably 1 or 2, and particularly preferably 1.

[0034] The anion of the organic ammonium salt of the present invention is not particularly limited, but examples thereof include carboxylate anions, halide anions, sulfur-based anions, fluorine-based anions, boron-based anions, nitrogen oxide-based anions, phosphorus-based anions, and cyanide-based anions.

[0035] The carboxylate anion has at least one carboxylate anion (-COO - ), and may contain a functional group having a heteroatom such as an oxygen atom, a nitrogen atom, or a sulfur atom. Examples of carboxylate anions include, but are not limited to, saturated aliphatic monocarboxylate anions, alicyclic carboxylate anions, unsaturated aliphatic monocarboxylate anions, saturated hydroxycarboxylate anions (saturated hydroxymonocarboxylate anions, saturated hydroxydi- or tricarboxylate anions, etc.), saturated dicarboxylate anions, unsaturated dicarboxylate anions, aromatic carboxylate anions, saturated carbonyl carboxylate anions, alkyl ether carboxylate anions, and halogen carboxylate anions (the number of carbon atoms in the carboxylate anions listed below includes the carbon atoms of the carboxy group).

[0036] The saturated aliphatic monocarboxylic acid anion comprises a linear or branched saturated aliphatic hydrocarbon group and one carboxylic acid anion, may contain a carboxy group or a carboxylate group, and preferably has 1 to 22 carbon atoms. Specific examples include anions formed by dissociating a proton from formic acid, acetic acid, propionic acid, butyric acid, valeric acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, lauric acid, myristic acid, pentadecylic acid, palmitic acid, margaric acid, stearic acid, arachidic acid, heneicosylic acid, behenic acid, isobutyric acid, 2-methylbutyric acid, isovaleric acid, 2-ethylhexanoic acid, isononanoic acid, isopalmitic acid, isostearic acid, etc.

[0037] When the anion of the organic ammonium salt is the saturated aliphatic monocarboxylic acid anion, an example of the compound of formula (I) is as follows: In formula (I), R is a linear or branched monohydroxyalkyl group having 1 to 10 carbon atoms, and n is an integer of 1 to 4. In formula (I), R is a linear monohydroxyalkyl group having 1 to 6 carbon atoms, and n is an integer of 1 to 4. In formula (I), R is a 1-hydroxyethyl group and n=1. In formula (I), R is a 1-hydroxyethyl group and n=2. In formula (I), R is a 1-hydroxyethyl group and n=3. R of formula (II) 11 is a hydrogen atom, a linear alkyl group having 1 to 4 carbon atoms, or a linear monohydroxyalkyl group having 1 to 4 carbon atoms, and n=1. In formula (I), R is a 1,3-dihydroxypropan-2-yl group and n=1. R of formula (II) 11 is a linear alkyl group having 1 to 4 carbon atoms, and n=1. In formula (I), R is a 1,3-dihydroxy-2-ethylpropan-2-yl group and n=1. R of formula (II) 11 is a linear monohydroxyalkyl group having 1 to 4 carbon atoms, and n=1. In formula (I), R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, and n=1. In formula (I), R is a linear or branched polyhydroxyalkyl group having 1 to 10 carbon atoms and two or more hydroxyl groups, and n is an integer of 1 to 4. In formula (I), R is a pentahydroxyhexan-1-yl group and n=1. In formula (I), R is a 1-hydroxyethyl group, n=3, and the saturated aliphatic monocarboxylic acid anion is a formate anion. In formula (I), R is a 1,3-dihydroxy-2-ethylpropan-2-yl group, n=1, or R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, n=1, and the saturated aliphatic monocarboxylic acid anion is an acetate anion. In formula (I), R is a 1,3-dihydroxypropan-2-yl group, n=1, R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, n=1, or R is a pentahydroxyhexan-1-yl group, n=1, and the saturated aliphatic monocarboxylic acid anion is a butyrate anion. In formula (I), R is a 1-hydroxyethyl group, n=1, R is a 1,3-dihydroxypropan-2-yl group, n=1, R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, n=1, or R is a pentahydroxyhexan-1-yl group, n=1, and the saturated aliphatic monocarboxylic acid anion is a caproate anion. In formula (I), R is a 1-hydroxyethyl group, n=1, R is a 1-hydroxyethyl group, n=3, R is a 1,3-dihydroxypropan-2-yl group, n=1, R is a 1,3-dihydroxypropan-2-yl group, n=1, R is a 1,3-dihydroxy-2-ethylpropan-2-yl group, n=1, R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, n=1, or R is a pentahydroxyhexan-1-yl group, n=1, and the saturated aliphatic monocarboxylic acid anion is caprylic acid anion. In formula (I), R is a 1-hydroxyethyl group, n=1, R is a 1-hydroxyethyl group, n=3, R is a 1,3-dihydroxypropan-2-yl group, n=1, R is a 1,3-dihydroxy-2-ethylpropan-2-yl group, n=1, R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, n=1, or R is a pentahydroxyhexan-1-yl group, n=1, and the saturated aliphatic monocarboxylic acid anion is capric acid anion. In formula (I), R is a 1-hydroxyethyl group, n=1, R is a 1-hydroxyethyl group, n=2, R is a 1-hydroxyethyl group, n=3, R is a 1,3-dihydroxypropan-2-yl group, n=1, R is a 1,3-dihydroxy-2-ethylpropan-2-yl group, n=1, R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, n=1, or R is a pentahydroxyhexan-1-yl group, n=1, and the saturated aliphatic monocarboxylic acid anion is a lauric acid anion.

[0038] The alicyclic carboxylate anion is composed of a non-aromatic saturated or unsaturated carbon ring and one or more carboxylate anions, and preferably has a carbon number of 6 to 20. Among these, alicyclic carboxylate anions having a cyclohexane ring skeleton are preferred, and specific examples include anions in which a proton is dissociated from cyclohexanecarboxylic acid and cyclohexanedicarboxylic acid.

[0039] When the anion of the organic ammonium salt is the above-mentioned alicyclic carboxylic acid anion, an example of the compound of formula (I) is as follows. In formula (I), R is a linear or branched monohydroxyalkyl group having 1 to 10 carbon atoms, and n is an integer of 1 to 4. In formula (I), R is a linear monohydroxyalkyl group having 1 to 6 carbon atoms, and n is an integer of 1 to 4. In formula (I), R is a 1-hydroxyethyl group and n=1. In formula (I), R is a 1-hydroxyethyl group and n=2. R of formula (II) 11 is a hydrogen atom, a linear alkyl group having 1 to 4 carbon atoms, or a linear monohydroxyalkyl group having 1 to 4 carbon atoms, and n=1. In formula (I), R is a 1,3-dihydroxypropan-2-yl group and n=1. In formula (I), R is a 1-hydroxyethyl group, n=1, R is a 1-hydroxyethyl group, n=3, or R is a 1,3-dihydroxypropan-2-yl group, n=1, and the alicyclic carboxylate anion is a cyclohexanecarboxylate anion.

[0040] The unsaturated aliphatic monocarboxylic acid anion comprises a linear or branched aliphatic unsaturated hydrocarbon group and one or more carboxylic acid anions, may contain a carboxy group or a carboxylate group, and preferably has a carbon number of 3 to 22. Specific examples include anions in which a proton is dissociated from acrylic acid, methacrylic acid, crotonic acid, palmitoleic acid, oleic acid, vaccenic acid, linoleic acid, linolenic acid, eleostearic acid, arachidonic acid, etc.

[0041] When the anion of the organic ammonium salt is the unsaturated aliphatic monocarboxylic acid anion, an example of the compound of formula (I) is as follows: In formula (I), R is a linear or branched monohydroxyalkyl group having 1 to 10 carbon atoms, and n is an integer of 1 to 4. In formula (I), R is a linear monohydroxyalkyl group having 1 to 6 carbon atoms, and n is an integer of 1 to 4. In formula (I), R is a 1-hydroxyethyl group and n=1. ·In formula (II), R 11 is a linear alkyl group having 1 to 4 carbon atoms, and n=1. In formula (I), R is a 1,3-dihydroxy-2-ethylpropan-2-yl group and n=1. In formula (I), R is a 1-hydroxyethyl group, n=1, and the unsaturated aliphatic monocarboxylic acid anion is a crotonate anion. In formula (I), R is a 1,3-dihydroxy-2-ethylpropan-2-yl group, and the unsaturated aliphatic monocarboxylic acid anion is an oleate anion.

[0042] The saturated hydroxycarboxylic acid anion comprises a linear or branched saturated aliphatic hydrocarbon group, one or more carboxylate anions, and one or more hydroxyl groups, and may contain a carboxyl group or a carboxylate group. It preferably has 2 to 24 carbon atoms. Among these, a saturated aliphatic hydroxycarboxylic acid anion having 2 to 7 carbon atoms and 1 to 4 hydroxyl groups is preferred. Specific examples include anions in which a proton is dissociated from glycolic acid, lactic acid, tartronic acid, glyceric acid, hydroxyacetic acid, hydroxybutyric acid, 2-hydroxydecanoic acid, 3-hydroxydecanoic acid, 12-hydroxystearic acid, dihydroxystearic acid, cerebronic acid, malic acid, tartaric acid, citramalic acid, citric acid, isocitric acid, leucic acid, mevalonic acid, pantoic acid, quinic acid, etc.

[0043] When the anion of the organic ammonium salt is a saturated aliphatic monohydroxycarboxylic acid anion among the saturated aliphatic hydroxycarboxylic acid anions, it preferably has 2 to 24 carbon atoms, more preferably 2 to 7 carbon atoms, and preferably has 1 to 4 hydroxyl groups. Examples of the compound of formula (I) are as follows: In formula (I), R is a linear or branched monohydroxyalkyl group having 1 to 10 carbon atoms, and n is an integer of 1 to 4. In formula (I), R is a linear monohydroxyalkyl group having 1 to 6 carbon atoms, and n is an integer of 1 to 4. In formula (I), R is a 1-hydroxyethyl group and n=1. In formula (I), R is a 1-hydroxyethyl group and n=2. In formula (I), R is a 1-hydroxyethyl group and n=3. ·In formula (II), R 11 is a hydrogen atom, a linear alkyl group having 1 to 4 carbon atoms, or a linear monohydroxyalkyl group having 1 to 4 carbon atoms, and n=1. In formula (I), R is a 1,3-dihydroxypropan-2-yl group and n=1. In formula (I), R is a 1,3-dihydroxy-2-ethylpropan-2-yl group and n=1. ·In formula (II), R 11 is a linear monohydroxyalkyl group having 1 to 4 carbon atoms, and n=1. In formula (I), R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, and n=1. In formula (I), R is a linear or branched polyhydroxyalkyl group having 1 to 10 carbon atoms and two or more hydroxyl groups, and n is an integer of 1 to 4. In formula (I), R is a pentahydroxyhexan-1-yl group and n=1. In formula (I), R is a 1,3-dihydroxy-2-ethylpropan-2-yl group, n=1, or R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, n=1, and the saturated aliphatic monohydroxycarboxylic acid anion is a glycolic acid anion. In formula (I), R is a 1,3-dihydroxypropan-2-yl group, n=1, R is a 1,3-dihydroxy-2-ethylpropan-2-yl group, n=1, R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, n=1, or R is a pentahydroxyhexan-1-yl group, n=1, and the saturated aliphatic monohydroxycarboxylic acid anion is a 9,10-dihydroxystearic acid anion. In formula (I), R is a 1-hydroxyethyl group, n=1, R is a 1-hydroxyethyl group, n=2, R is a 1,3-dihydroxypropan-2-yl group, n=1, or R is a 1,3-dihydroxy-2-ethylpropan-2-yl group, n=1, and the saturated aliphatic monohydroxycarboxylic acid anion is a quinate anion. In formula (I), R is a 1,3-dihydroxypropan-2-yl group, n=1, and the saturated aliphatic monohydroxycarboxylic acid anion is a 12-hydroxystearic acid anion.

[0044] When the anion of the organic ammonium salt is a saturated aliphatic hydroxy di- or tricarboxylic acid anion among the saturated aliphatic hydroxy carboxylic acid anions, it preferably has 4 to 24 carbon atoms and preferably has 1 to 3 hydroxyl groups. Examples of the compound of formula (I) are as follows. In formula (I), R is a linear or branched monohydroxyalkyl group having 1 to 10 carbon atoms, and n is an integer of 1 to 4. In formula (I), R is a linear monohydroxyalkyl group having 1 to 6 carbon atoms, and n is an integer of 1 to 4. In formula (I), R is a 1-hydroxyethyl group and n=1. In formula (I), R is a 1-hydroxyethyl group, n=1, and the saturated aliphatic hydroxy di- or tricarboxylic acid anion is tartaric acid. In formula (I), R is a 1-hydroxyethyl group, n=1, and the saturated aliphatic hydroxy di- or tricarboxylic acid anion is a citrate anion.

[0045] The saturated dicarboxylic acid anion preferably has 2 to 24 carbon atoms, more preferably 2 to 10 carbon atoms. Specific examples include anions in which a proton is dissociated from oxalic acid, malonic acid, succinic acid, glutaric acid, adipic acid, pimelic acid, suberic acid, azelaic acid, sebacic acid, etc.

[0046] When the anion of the organic ammonium salt is the saturated dicarboxylic acid anion, an example of the compound of formula (I) is as follows: In formula (I), R is a linear or branched monohydroxyalkyl group having 1 to 10 carbon atoms, and n is an integer of 1 to 4. In formula (I), R is a linear monohydroxyalkyl group having 1 to 6 carbon atoms, and n is an integer of 1 to 4. In formula (I), R is a 1-hydroxyethyl group and n=1. In formula (I), R is a 1-hydroxyethyl group and n=3. ·In formula (II), R 11 is a hydrogen atom, a linear alkyl group having 1 to 4 carbon atoms, or a linear monohydroxyalkyl group having 1 to 4 carbon atoms, and n=1. In formula (I), R is a 1,3-dihydroxypropan-2-yl group and n=1. ·In formula (II), R 11 is a linear monohydroxyalkyl group having 1 to 4 carbon atoms, and n=1. In formula (I), R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, and n=1. In formula (I), R is a 1,3-dihydroxypropan-2-yl group, n=1, and the saturated aliphatic monohydroxycarboxylic acid anion is an oxalate anion. In formula (I), R is a 1-hydroxyethyl group, n=1, or R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, n=1, and the dicarboxylic acid anion is a succinate anion. In formula (I), R is a 1-hydroxyethyl group, n=1, R is a 1,3-dihydroxypropan-2-yl group or n=1, or a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, n=1, and the adipate anion is an adipate anion. In formula (I), R is a 1,3-dihydroxypropan-2-yl group, and the dicarboxylate anion is a sebacate anion.

[0047] The unsaturated dicarboxylic acid anion preferably has 2 to 24 carbon atoms, more preferably 2 to 10 carbon atoms. Specific examples include anions in which a proton is dissociated from maleic acid, fumaric acid, citraconic acid, mesaconic acid, 2-pentenedioic acid, methylenesuccinic acid, allylmalonic acid, isopropylidesuccinic acid, adipic acid, 2,4-hexadienedioic acid, etc.

[0048] When the anion of the organic ammonium salt is the unsaturated dicarboxylic acid anion, an example of the compound of formula (I) is as follows: In formula (I), R is a linear or branched monohydroxyalkyl group having 1 to 10 carbon atoms, and n is an integer of 1 to 4. In formula (I), R is a linear monohydroxyalkyl group having 1 to 6 carbon atoms, and n is an integer of 1 to 4. In formula (I), R is a 1,3-dihydroxypropan-2-yl group and n=1. In formula (I), R is a 1,3-dihydroxypropan-2-yl group, n=1, and the dicarboxylic acid anion is a maleate anion.

[0049] The aromatic carboxylate anion contains a single ring or multiple rings having aromaticity and one or more carboxylate anions, and preferably has a carbon number of 6 to 20. Among these, aromatic carboxylate anions having a benzene ring skeleton are preferred, and specific examples include anions in which a proton is dissociated from benzoic acid, cinnamic acid, phthalic acid, isophthalic acid, terephthalic acid, etc.

[0050] Among the aromatic carboxylate anions, the aromatic hydroxycarboxylate anion is composed of an aromatic single ring or multiple rings, one or more carboxylate anions, and one or more hydroxyl groups, and preferably has 6 to 20 carbon atoms. Of these, aromatic carboxylate anions having a benzene ring skeleton having 1 to 3 hydroxyl groups are preferred, and specific examples include anions obtained by dissociating a proton from salicylic acid, hydroxybenzoic acid, dihydroxybenzoic acid, trihydroxybenzoic acid, hydroxymethylbenzoic acid, vanillic acid, syringic acid, pyrotocatechuic acid, gentisic acid, orselliic acid, mandelic acid, benzilic acid, atrolactic acid, phloretic acid, coumaric acid, umbellic acid, caffeic acid, ferulic acid, sinapic acid, etc.

[0051] When the anion of the organic ammonium salt is the aromatic carboxylic acid anion, an example of the compound of formula (I) is as follows: In formula (I), R is a linear or branched monohydroxyalkyl group having 1 to 10 carbon atoms, and n is an integer of 1 to 4. In formula (I), R is a linear monohydroxyalkyl group having 1 to 6 carbon atoms, and n is an integer of 1 to 4. In formula (I), R is a 1-hydroxyethyl group and n=1. In formula (I), R is a 1-hydroxyethyl group and n=2. In formula (I), R is a 1-hydroxyethyl group and n=3. ·In formula (II), R 11 is a hydrogen atom, a linear alkyl group having 1 to 4 carbon atoms, or a linear monohydroxyalkyl group having 1 to 4 carbon atoms, and n=1. In formula (I), R is a 1,3-dihydroxypropan-2-yl group and n=1. ·In formula (II), R 11 is a linear alkyl group having 1 to 4 carbon atoms, and n=1. In formula (I), R is a 1,3-dihydroxy-2-ethylpropan-2-yl group and n=1. ·In formula (II), R 11 is a linear monohydroxyalkyl group having 1 to 4 carbon atoms, and n=1. In formula (I), R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, and n=1. In formula (I), R is a linear or branched polyhydroxyalkyl group having 1 to 10 carbon atoms and two or more hydroxyl groups, and n is an integer of 1 to 4. In formula (I), R is a pentahydroxyhexan-1-yl group and n=1. In formula (I), R is a 1-hydroxyethyl group, n=1, R is a 1-hydroxyethyl group, n=3, R is a 1,3-dihydroxypropan-2-yl group, n=1, R is a 1,3-dihydroxy-2-ethylpropan-2-yl group, n=1, R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, n=1 or a pentahydroxyhexan-1-yl group, n=1, and the aromatic carboxylate anion is a benzoate anion. In formula (I), R is a 1-hydroxyethyl group, n=1, R is a 1-hydroxyethyl group, n=2, R is a 1-hydroxyethyl group, n=3, or R is a 1,3-dihydroxypropan-2-yl group, n=1, and the aromatic carboxylate anion is a terephthalate anion. In formula (I), R is a 1-hydroxyethyl group, n=1, R is a 1,3-dihydroxy-2-ethylpropan-2-yl group, n=1, or R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, n=1, and the aromatic carboxylate anion is a salicylate anion. In formula (I), R is a 1-hydroxyethyl group, n=1, R is a 1-hydroxyethyl group, n=3, or R is a 1,3-dihydroxypropan-2-yl group, n=1, and the aromatic carboxylate anion is a p-hydroxybenzoate anion. In formula (I), R is a 1-hydroxyethyl group, n=1, and the aromatic carboxylate anion is a mandelate anion.

[0052] The saturated carbonyl carboxylate anion is a carboxylate anion having 3 to 22 carbon atoms and a carbonyl group in the molecule, and is preferably a saturated carbonyl carboxylate anion having 3 to 7 carbon atoms and one or two carbonyl groups. Among these, CH3((CH2) pCO(CH2) q )COO - (p and q are integers of 0 to 2.) A specific example of the anion is an anion in which a proton is dissociated from pyruvic acid or the like.

[0053] When the anion of the organic ammonium salt is the saturated carbonyl carboxylic acid anion, an example of the compound of formula (I) is as follows: In formula (I), R is a linear or branched polyhydroxyalkyl group having 1 to 10 carbon atoms and two or more hydroxyl groups, and n is an integer of 1 to 4. In formula (I), R is a pentahydroxyhexan-1-yl group and n=1. In formula (I), R is a pentahydroxyhexan-1-yl group, n=1, and the saturated carbonyl carboxylate anion is a pyruvate anion.

[0054] The alkyl ether carboxylate anion is a carboxylate anion having 2 to 22 carbon atoms and an ether group in the molecule, including a polyoxyalkylene alkyl ether carboxylate anion, and an alkyl carboxylate anion having 2 to 12 carbon atoms and one or two ether groups is preferred. Among these, CH3(CH2) r O(CH2) s COO - (r and s are integers of 0 to 4.) Preferred are alkyl ether carboxylate anions and polyoxyethylene alkyl ether carboxylate anions represented by the following formula: (r and s are integers of 0 to 4.) Specific examples include anions in which a proton is dissociated from methoxyacetic acid, ethoxyacetic acid, methoxybutyric acid, ethoxybutyric acid, etc.

[0055] When the anion of the organic ammonium salt is the alkyl ether carboxylic acid anion, an example of the compound of formula (I) is as follows: ·In formula (II), R 11 is a linear alkyl group having 1 to 4 carbon atoms, and n=1 In formula (I), R is a 1,3-dihydroxy-2-ethylpropan-2-yl group and n=1. In formula (I), R is a 1,3-dihydroxy-2-ethylpropan-2-yl group, n=1, and the alkyl ether carboxylic acid anion is a methoxyacetate anion.

[0056] The halogen carboxylic acid anion is preferably a halogen carboxylic acid anion having 2 to 22 carbon atoms. Specific examples include anions in which a proton is dissociated from a fluorine-substituted halogen carboxylic acid such as trifluoroacetic acid, trichloroacetic acid, tribromoacetic acid, pentafluoropropionic acid, pentachloropropionic acid, pentabromopropionic acid, perfluorononanoic acid, perchlorononanoic acid, or perbromononanoic acid.

[0057] When the anion of the organic ammonium salt is the halogen carboxylate anion, an example of the compound of formula (I) is as follows: In formula (I), R is a linear or branched monohydroxyalkyl group having 1 to 10 carbon atoms, and n is an integer of 1 to 4. In formula (I), R is a linear monohydroxyalkyl group having 1 to 6 carbon atoms, and n is an integer of 1 to 4. In formula (I), R is a 1-hydroxyethyl group and n=1. ·In formula (II), R 11 is a linear alkyl group having 1 to 4 carbon atoms, and n=1. In formula (I), R is a 1,3-dihydroxy-2-ethylpropan-2-yl group and n=1. ·In formula (II), R 11 is a linear monohydroxyalkyl group having 1 to 4 carbon atoms, and n=1. In formula (I), R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, and n=1. In formula (I), R is a 1-hydroxyethyl group, n=1, R is a 1,3-dihydroxy-2-ethylpropan-2-yl group, n=1, or R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, n=1, and the halogen carboxylate anion is a trifluoroacetate anion.

[0058] The halide anion is not particularly limited, but examples thereof include chloride ion, bromide ion, and iodide ion.

[0059] When the anion of the organic ammonium salt is the above-mentioned halide anion, an example of the compound of formula (I) is as follows: In formula (I), R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, and n=1. In formula (I), R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, n=1, and the halide anion is a bromide ion. In formula (I), R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, n=1, and the halide anion is a chloride ion.

[0060] Examples of the sulfur-based anion include sulfate anion, sulfite anion, sulfonate anion, hydrogen sulfonate anion, alkyl sulfonate anion (e.g., methanesulfonate, ethyl sulfonate, butyl sulfonate, benzenesulfonate, p-toluenesulfonate, 2,4,6-trimethylbenzenesulfonate, styrenesulfonate, 3-sulfopropyl methacrylate anion, 3-sulfopropyl acrylate, etc.), sulfate anion, hydrogen sulfate anion, alkyl sulfate anion (e.g., methyl sulfate anion, ethyl sulfate anion, butyl sulfate anion, octyl sulfate anion, 2-(2-methoxyethoxy)ethyl sulfate anion, etc.).

[0061] When the anion of the organic ammonium salt is the above-mentioned sulfur-based anion, an example of the compound of formula (I) is as follows. ·In formula (II), R 11 is a linear monohydroxyalkyl group having 1 to 4 carbon atoms, and n=1. In formula (I), R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, and n=1. In formula (I), R is a linear or branched polyhydroxyalkyl group having 1 to 10 carbon atoms and two or more hydroxyl groups, and n is an integer of 1 to 4. In formula (I), R is a pentahydroxyhexan-1-yl group and n=1. In formula (I), R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, n=1, or R is a pentahydroxyhexan-1-yl group, n=1, and the sulfur-based anion is a sulfate anion.

[0062] Examples of the fluorine-based anion include bis(fluorosulfonyl)imide anion, bis(perfluoroalkylsulfonyl)imide anion (e.g., bis(trifluoromethylsulfonyl)imide anion, bis(pentafluoroethylsulfonyl)imide, bis(heptafluoropropanesulfonyl)imide anion, bis(nonafluorobutylsulfonyl)imide, etc.), perfluoroalkylsulfonate anion (e.g., trifluoromethanesulfonate anion, pentafluoroethanesulfonate anion, heptafluoropropanesulfonate anion, nonaflate anion, etc.), and perfluorooctanesulfonate anion, etc.), fluorophosphate anions (for example, hexafluorophosphate anion, tri(pentafluoroethyl)trifluorophosphate anion, etc.), tris(perfluoroalkylsulfonyl)methide anions (for example, tris(trifluoromethanesulfonyl)methide anion, tris(pentafluoroethanesulfonyl)methide anion, tris(heptafluoropropanesulfonyl)methide anion, tris(nonafluorobutanesulfonyl)methide anion, etc.), fluorohydrogenate anion, etc.

[0063] When the anion of the organic ammonium salt is the above-mentioned fluorine-based anion, an example of the compound of formula (I) is as follows. ·In formula (II), R 11 is a hydrogen atom, a linear alkyl group having 1 to 4 carbon atoms, or a linear monohydroxyalkyl group having 1 to 4 carbon atoms, and n=1. In formula (I), R is a 1,3-dihydroxypropan-2-yl group and n=1.

[0064] In formula (I), R is a 1,3-dihydroxypropan-2-yl group, n=1, and the fluorine-based anion is a bis(trifluoromethylsulfonyl)imide anion.

[0065] The boron-based anion is not particularly limited, but examples thereof include tetrafluoroborate anion, bisoxalatoborate anion, and tetraalkylborate anion such as tetraphenylborate anion.

[0066] The nitrogen oxide anion is not particularly limited, but examples thereof include nitrate anion and nitrite anion.

[0067] Examples of the phosphorus-based anion include phosphate anion, hydrogen phosphate anion, dihydrogen phosphate anion, phosphonate anion, hydrogen phosphonate anion, dihydrogen phosphonate anion, phosphinate anion, hydrogen phosphinate anion, alkyl phosphate anion (e.g., dimethyl phosphate, diethyl phosphate, dipropyl phosphate anion, dibutyl phosphate anion, etc.), alkyl phosphonate anion (e.g., methyl phosphonate anion, ethyl phosphonate anion, propyl phosphonate anion, butyl phosphonate anion, methylmethyl phosphonate anion, etc.), alkyl phosphinate anion, hexaalkyl phosphate anion, etc.

[0068] When the anion of the organic ammonium salt is the above phosphorus-based anion, an example of the compound of formula (I) is as follows: ·In formula (II), R 11 is a linear monohydroxyalkyl group having 1 to 4 carbon atoms, and n=1. In formula (I), R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, and n=1. In formula (I), R is a linear or branched polyhydroxyalkyl group having 1 to 10 carbon atoms and two or more hydroxyl groups, and n is an integer of 1 to 4. In formula (I), R is a pentahydroxyhexan-1-yl group and n=1.

[0069] In formula (I), R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, n=1, or R is a pentahydroxyhexan-1-yl group, n=1, and the phosphorus-based anion is a dihydrogen phosphate anion.

[0070] The cyanide-based anion is not particularly limited, but examples thereof include tetracyanoborate anion, dicyanamide anion, thiocyanate anion, and isothiocyanate anion.

[0071] When the anion of the organic ammonium salt is the above-mentioned cyanide-based anion, an example of the compound of formula (I) is as follows. ·In formula (II), R 11 is a hydrogen atom, a linear alkyl group having 1 to 4 carbon atoms, or a linear monohydroxyalkyl group having 1 to 4 carbon atoms, and n=1. In formula (I), R is a 1,3-dihydroxypropan-2-yl group and n=1. ·In formula (II), R 11 is a linear monohydroxyalkyl group having 1 to 4 carbon atoms, and n=1. In formula (I), R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, and n=1. In formula (I), R is a linear or branched polyhydroxyalkyl group having 1 to 10 carbon atoms and two or more hydroxyl groups, and n is an integer of 1 to 4. In formula (I), R is a pentahydroxyhexan-1-yl group and n=1. In formula (I), R is a 1,3-dihydroxypropan-2-yl group, n=1, R is a 1,3-dihydroxy-2-hydroxymethylpropan-2-yl group, n=1, or R is a pentahydroxyhexan-1-yl group, n=1, and the cyanide-based anion is a dicyanamide anion.

[0072] Of the anions of the organic ammonium salt of the present invention, carboxylate anions, halide anions, sulfur-based anions, fluorine-based anions, nitrogen oxide-based anions, phosphorus-based anions, and cyanide-based anions are preferred embodiments, and carboxylate anions are particularly preferred from the viewpoint of safety, and among these, saturated aliphatic monocarboxylate anions, alicyclic carboxylate anions, unsaturated aliphatic monocarboxylate anions, saturated hydroxycarboxylate anions, saturated dicarboxylate anions, unsaturated dicarboxylate anions, saturated hydroxydi- or tricarboxylate anions, aromatic carboxylate anions, saturated carbonyl carboxylate anions, alkyl ether carboxylate anions, and halogen carboxylate anions are preferred embodiments. The organic ammonium salt of the present invention is not particularly limited, but can be synthesized, for example, as follows.

[0073] An alkanolamine having one or more hydroxy groups corresponding to R in formula (I), an amino acid having one or more carboxy groups, or an aminohydroxyalkanoic acid having one or more hydroxy groups and one or more carboxy groups is reacted with an organic acid or inorganic acid corresponding to the anion in a solvent such as water or an organic solvent. Alternatively, an alkanolamine having one or more hydroxy groups corresponding to R in formula (I), an amino acid having one or more carboxy groups, or an aminohydroxyalkanoic acid having one or more hydroxy groups and one or more carboxy groups is reacted with an organic halogen compound such as an alkylene halohydrin, a monohaloalkylcarboxylic acid, or a monohalohydroxyalkylcarboxylic acid in a solvent, and the resulting compound is reacted with an organic acid or inorganic acid corresponding to the anion of the target compound in a solvent such as water or an organic solvent.

[0074] The reaction is carried out in a polar solvent such as water or acetonitrile with an alkanolamine (e.g., mono-, di-, or trialkanolamine, 2-amino-1,3-propanediol, 2-amino-2-ethyl-1,3-propanediol, tris(hydroxymethyl)aminomethane, or D-glucamine) corresponding to the ammonium cation of formula (I), an amino acid (e.g., glycine, aspartic acid, or glutamic acid) containing a carboxyalkyl group, or an aminohydroxyalkanoic acid (e.g., 3-amino-2-hydroxypropionic acid) containing at least one hydroxyl group and at least one carboxyl group, and an organic or inorganic acid corresponding to the anion. The reaction temperature and reaction time vary depending on the raw materials, but can be carried out at room temperature for approximately one hour to one day. The solvent is then removed by distillation under reduced pressure, and the desired organic ammonium salt is obtained as a solid by purification, if necessary. Furthermore, if the reaction is completed with equimolar amounts, a purification step is not required, further simplifying the production process.

[0075] Furthermore, a compound of formula (I) in which R is composed of a hydroxycarboxyalkyl group, a hydroxyalkyl group, or a carboxyalkyl group and has no hydrogen atoms (n is 4) can also be synthesized, for example, as follows.

[0076] In the first step, an aminohydroxyalkanoic acid (e.g., 3-amino-2-hydroxypropionic acid) having at least one hydroxyl group and at least one carboxyl group is reacted with a hydroxyalkyl halide having at least two hydroxyl groups or a haloalkylcarboxylic acid having at least two carboxyl groups in a polar solvent such as water or acetonitrile to produce an ammonium cation of formula (I). The reaction temperature and reaction time vary depending on the types of raw materials, but can be carried out, for example, at room temperature for about one day. The reaction product is then washed to obtain a compound consisting of the ammonium cation of formula (I) and a halide ion. To further convert the halide ion into the desired anion, anion exchange is performed. For anion exchange, the resulting compound is reacted in water with an organic or inorganic acid corresponding to the anion of the desired compound. The reaction temperature and reaction time vary depending on the types of raw materials, but can be carried out, for example, at room temperature for about one day. Alternatively, the desired organic ammonium salt can be obtained by anion-exchanging the compound to hydroxide anions using a strongly basic ion-exchange resin or the like, followed by further anion-exchanging with an organic acid or inorganic acid corresponding to the anion of the desired compound.

[0077] The organic ammonium salt of the present invention is solid at 25°C. Here, "solid" refers to a state in which the salt is non-flowing at 25°C, including both crystalline and amorphous salts. The melting point (freezing point) may vary depending on the purity of the organic ammonium salt, but this term encompasses compounds of formula (I) that are solid at 25°C when synthesized in consideration of conventional technical knowledge. The purity of the organic ammonium salt may vary depending on the purification method and the molar ratio of anions to cations in the product. However, the melting point (freezing point) of the organic ammonium salt synthesized and further purified as described in the Examples of the present invention is highly pure as determined by IR and NMR spectroscopy, and has a melting point uniquely determined by the compound itself, so the results of the Examples of the present invention are to be referenced. The organic ammonium salt of the present invention may be in an anhydrous state (anhydride) or a hydrate that has absorbed moisture from the air. A hydrate refers to a compound that absorbs water and reaches a saturated moisture content when left in air at 25°C. A compound that does not absorb water when left in air at 25°C is an anhydrous compound.

[0078] By selecting the functional group or characteristic group of the ammonium cation and the anion, the organic ammonium salt of the present invention is solid at 25°C and encompasses ionic liquids, which are organic salts with a melting point of 100°C or less in a broad sense. When reacting at high temperatures, ionic liquids are nonvolatile due to their structural characteristics and have a high decomposition temperature. For example, they can react at temperatures above 100°C, which is higher than that of water as a solvent, expanding the applicable range of reaction temperatures. In addition, they are highly convenient because of their very low flammability and combustibility.

[0079] The organic ammonium salt of the present invention has high affinity with hydrogen-bonding materials having a hydrogen-bonding functional group or a hydrogen-bond-accepting element that interacts with a hydroxy group, carboxy group, or hydrogen atom bonded to nitrogen of the cation, and can be used in a variety of applications requiring affinity with hydrogen-bonding materials capable of forming hydrogen bonds, coordinate bonds, or ionic bonds.

[0080] The hydrogen atoms bonded to the hydrogen or nitrogen atoms of the hydroxyl group or carboxyl group of the organic ammonium salt of the present invention form hydrogen bonds with, but are not limited to, elements having lone electron pairs such as those of Groups 15 to 17, or π-electron compounds, or the oxygen atoms of the hydroxyl group or carboxyl group of the organic ammonium salt of the present invention form hydrogen bonds with hydrogen atoms of the target compound or material, and therefore have excellent affinity with compounds or materials having hydrogen-bonding functional groups.

[0081] Furthermore, since the organic ammonium salt of the present invention is an organic salt having a hydroxyl group and / or a carboxyl group, it has excellent affinity with metals, ionic compounds, and materials due to coordination interactions and electrostatic interactions. The metal is not particularly limited, but examples thereof include elements of Groups 2 to 14.

[0082] Examples of the hydrogen-bonding functional group include a carbon-carbon unsaturated bond group, an oxygen-containing group, a nitrogen-containing group, a sulfur-containing group, a phosphorus-containing group, and a hydrogen atom directly bonded to nitrogen. The carbon-carbon unsaturated bond group is not particularly limited, but examples thereof include a vinyl group, a vinylene group, an ethynyl group, and an unsaturated cyclic hydrocarbon group.

[0083] The oxygen-containing group is not particularly limited, but examples thereof include a hydroxyl group, a carbonyl group, an ether group, an ester group, an aldehyde group, a carboxyl group, a carboxylate group, a urea group, a urethane group, an amide group, an oxazole group, a morpholine group, a carbamic acid group, and a carbamate group.

[0084] The nitrogen-containing group is not particularly limited, but examples thereof include an amino group and a nitro group.

[0085] The sulfur-containing group is not particularly limited, and examples thereof include a sulfate group (-OS(=O)2O-), a sulfonyl group (-S(=O)2O-), a sulfonic acid group (-S(=O)2-), a mercapto group (-SH), a thioether group (-S-), a thiocarbonyl group (-C(=S)-), a thiourea group (-NC(=S)-N-), a thiocarboxyl group (-C(=S)OH), a thiocarboxylate group (-C(=S)O-), a dithiocarboxyl group (-C(=S)SH), and a dithiocarboxylate group (-C(=S)S-).

[0086] The phosphorus-containing group is not particularly limited, and examples thereof include a phosphate group (-OP(=O)(-O-)-O-), a phosphonate group (-P(=O)(-O-)-O-), a phosphinate group (-P(=O)-O-), a phosphorous acid group (-OP(-O-)-O-), a phosphonous acid group (-P(-O-)-O-), a phosphinous acid group (-PO-), a pyrophosphate group [(-OP(=O)(-O-))2-O-], and the like.

[0087] Examples of hydrogen-bonding materials having a hydrogen-bond-accepting element include materials containing compounds having a hydrogen-bond-accepting element or an oxide thereof as a constituent element of a molecule or functional group, and materials whose surfaces are modified with functional groups containing such elements or oxides thereof.

[0088] The organic ammonium salt of the present invention has a hydrogen-bonding functional group, such as a hydroxyl group, a carboxyl group, or a hydrogen atom bonded to a nitrogen atom, in the cation, and therefore has excellent affinity with hydrogen-bonding materials. Furthermore, if the anion of the organic ammonium salt also has a hydrogen-bonding functional group, the affinity with hydrogen-bonding materials will be even better. The hydrogen-bonding material is not particularly limited, but examples thereof include biological samples and organic or inorganic compound materials.

[0089] Examples of biological samples include biocatalysts such as enzymes, peptides, proteins, nucleic acids, poorly soluble polysaccharides such as cellulose, cells, cell tissue fluid, cell membranes, blood, biological tissues, antibodies, antigens, and the like.

[0090] Examples of organic compound materials include compounds having the above-mentioned hydrogen-bonding functional groups, including, but not limited to, organic resins, organic pigments, organic dyes, and organic fluorescent dyes. Examples of organic resins include, but are not limited to, thermoplastic resins and thermosetting resins. Examples of organic pigments include, but are not limited to, azo-based, diazo-based, condensed azo-based, thioindigo-based, indathrone-based, quinacridone-based, anthraquinone-based, benzimidazolone-based, phenylene-based, phthalocyanine-based, anthrapyridine-based, and dioxazine-based. Examples of organic fluorescent dyes include, but are not limited to, rhodamine-based dyes, squarylium-based dyes, cyanine-based dyes, aromatic hydrocarbon-based dyes, oxazine-based dyes, carbopyronine-based dyes, and pyrromethene-based dyes. Examples of organic dyes include, but are not limited to, organic solvent-soluble dyes classified as solvent dyes in the Color Index. Specific examples of solvent dyes include Varifast Black 3806, Varifast Black 3807, Varifast Black 3830, Spirit Black SB, Spiron Black GMH, Varifast Red 1320, Varifast Red 1308, Varifast Yellow AUM, Spiron Yellow C2GH, Spiron Violet CRH, Varifast Violet 1701, Spiron Red CGH, Spiron Pink BH, Nigrosine Base EX, Oil Blue 613, Neozapon Blue 808, and Varifast Blue 1621.

[0091] The inorganic compound material is not particularly limited, but examples thereof include metals, metal oxides, rare earth metal oxides, hydroxides, carbonates, sulfates, silicates, nitrides, titanic acid compounds, carbons, etc. The metal is not particularly limited, but examples thereof include elements of Groups 2 to 14, such as iron, aluminum, chromium, nickel, cobalt, zinc, tungsten, indium, tin, palladium, zirconium, titanium, copper, silver, gold, platinum, etc. Metal oxides are preferably used because they form good hydrogen bonds with the organic ammonium salt of the present invention. Examples of metal oxides include, but are not limited to, silica, aluminum oxide (alumina), zirconia, titanium oxide, magnesium oxide, indium tin oxide (ITO), cobalt blue (CoO·Al2O3), antimony oxide, zinc oxide, cesium oxide, zirconium oxide, yttrium oxide, tungsten oxide, vanadium oxide, cadmium oxide, tantalum oxide, niobium oxide, tin oxide, bismuth oxide, cerium oxide, copper oxide, iron oxide, indium oxide, boron oxide, calcium oxide, barium oxide, thorium oxide, indium tin oxide, and ferrite. Examples of rare earth metal oxides include dysprosium oxide, erbium oxide, europium oxide, gadolinium oxide, holmium oxide, lanthanum oxide, lutetium oxide, neodymium oxide, praseodymium oxide, samarium oxide, scandium oxide, terbium oxide, thulium oxide, and ytterbium oxide. Examples of hydroxides include calcium hydroxide, magnesium hydroxide, aluminum hydroxide, basic magnesium carbonate, iron hydroxide, etc. Examples of carbonates include calcium carbonate, magnesium carbonate, zinc carbonate, barium carbonate, dawsonite, hydrotalcite, etc. Examples of sulfates include calcium sulfate, barium sulfate, aluminum sulfate, etc.Examples of silicates include calcium silicate, wollastonite, xonotlite, kaolin, talc, clay, mica, montmorillonite, bentonite, dolomite, hydrotalcite, calcium silicate, aluminum silicate, magnesium silicate, zirconium silicate, activated clay, sepiolite, imogolite, sericite, glass fiber, glass beads, and silica-based balloons. Examples of nitrides include aluminum nitride, boron nitride, and silicon nitride. Examples of titanate compounds include barium titanate, barium titanate zirconate, calcium titanate, and strontium titanate. Examples of carbons include carbon black, graphite, carbon fiber, carbon balloons, activated carbon, bamboo charcoal, charcoal, single-walled carbon nanotubes, double-walled carbon nanotubes, multi-walled carbon nanotubes, carbon nanohorns, and fullerenes.

[0092] (Hydrogen-bonding material treatment agent) The organic ammonium salt of the present invention can be used as a processing agent for organic or inorganic hydrogen-bonding materials. For example, it is expected to be used in dispersion media for hydrogen-bonding materials, biological sample preservation materials, biological sample processing agents such as protein refolding agents, moisture regulators such as antifogging agents and humidity conditioners, surface treatment agents such as dispersants for inorganic and organic materials, electrolyte materials, conductive materials, electronic materials such as fuel cells, resin additives such as antistatic agents, life science materials such as pharmaceuticals, cosmetics, toiletries, antibacterial agents, disinfectants, and deodorizers, industrial materials such as reaction aids, heat transfer media, adhesives, antifogging agents, adsorbents, heat insulating agents, and polymer substrates, agricultural materials such as agricultural sustained-release agents, etc.

[0093] The hydrogen-bonding material treating agent of the present invention contains the organic ammonium salt of the present invention described above. Here, "contains" refers primarily to the organic ammonium salt of the present invention, but may also contain other optional components that are acceptable depending on the intended use. The optional components are not particularly limited, but examples include solvents and dispersion media such as water and organic solvents, and these can be used as solutions or dispersions. Among the solvents and dispersion media, water, alcohol, and the like, which have a high affinity with the organic ammonium salt of the present invention, are preferably used.

[0094] The organic ammonium salt of the present invention can stably store and disperse a hydrogen-bonding material in the form of a solution or dispersion obtained by mixing the hydrogen-bonding material with water or a solvent. Even if water or solvent is removed from the solution or dispersion to form a solid composition, and then a solvent is added again to form a solution or dispersion, the resulting composition can be stably stored, dissolved, and dispersed.

[0095] The hydrogen-bonding material treatment agent containing the organic ammonium salt of the present invention is mixed with a hydrogen-bonding material and a solvent to form a solution or dispersion, and then the solvent is removed to obtain a solid composition in which the hydrogen-bonding material is uniformly mixed in the hydrogen-bonding treatment agent. This is useful for storing the hydrogen-bonding material in a solid state. Furthermore, adding a solvent to the solid composition again produces a solution or dispersion in which the hydrogen-bonding material is uniformly dissolved or dispersed, making it useful as a method for uniformly dissolving or dispersing the hydrogen-bonding material again after it has been solidified.

[0096] (biological sample treatment agent) The biological sample treating agent is one of the above-mentioned hydrogen-bonding material treating agents for treating biological samples, and examples thereof include biological sample preservation materials, protein refolding agents, stabilizers for biosensors, surface treating agents for biological samples, modifiers for biological samples, hair treatment agents, skin care agents, etc., as described above.

[0097] The following mainly describes agents for treating hydrogen-bonding materials, particularly agents for treating biological samples. However, an explanation of agents for treating hydrogen-bonding materials is also included, and the following description is also referred to for details of biological samples and the effects of the organic ammonium salt of the present invention on various treating agents based on their affinity for the target hydrogen-bonding material and their interaction with hydrogen-bonding functional groups.

[0098] The hydrogen-bonding material treating agent of the present invention can be used as a biological sample treating agent, and can be used, for example, to maintain the three-dimensional structure of a biological sample, activate a biological sample, maintain the activity of a biological sample, store the biological sample for a long period while maintaining the activity, etc. Therefore, it is useful, for example, for refolding proteins, maintaining the activity of enzymes, and storing a biological sample while maintaining the three-dimensional structure, etc.

[0099] The biological sample treating agent of the present invention contains the organic ammonium salt of the present invention described above. Here, "containing" refers primarily to an agent consisting of the organic ammonium salt of the present invention, but may also contain other optional components that are acceptable for treating biological samples.

[0100] Examples of biological samples include biocatalysts such as enzymes, peptides, proteins, nucleic acids, poorly soluble polysaccharides such as cellulose, cells, cell tissue fluid, cell membranes, blood, biological tissues, antibodies, antigens, etc. Among these, biocatalysts, proteins, and nucleic acids are preferred.

[0101] Among biological samples, biocatalysts are catalysts for biochemical reactions, and biocatalysts in this invention include microorganisms, animal and plant cells, tissues, and enzymes derived from these organisms, as well as artificial compounds with enzymatic functions and artificial enzymes that have been artificially modified from naturally occurring enzymes or biomolecules to give them new properties.

[0102] Enzymes have a primary structure formed by one-dimensional binding of amino acids, but the arrangement and number of amino acids determine the two-dimensional and higher-dimensional structures. These structures determine the unique properties of each enzyme.

[0103] The primary structure is a linear arrangement of 20 amino acids via peptide bonds. Most enzymes consist of 100–300 amino acids, and the order of these amino acids determines the enzyme's properties. The secondary structure is a structure in which certain portions of the overall linear sequence have highly ordered structures such as α-helices, β-sheets, and β-turns. The tertiary structure is a three-dimensional structure formed by the primary and secondary structures. This three-dimensional structure determines the active center where the enzyme acts as a catalyst and the three-dimensional structure of amino acid residues consisting of hydrophilic and hydrophobic regions. This structure allows enzymes and other biocatalysts to exhibit unique substrate and reaction specificities that are not found in general proteins (structural proteins, transport proteins, storage proteins, contractile proteins, defense proteins, and hormone proteins). The quaternary structure is an assembly of multiple enzyme molecules in a three-dimensional structure. In other words, enzymes and other biocatalysts possess reaction specificity based on their primary, secondary, and quaternary structures in addition to their substrate specificity. Therefore, maintaining not only the primary and secondary structures but also the tertiary and quaternary structures is important for maintaining catalytic activity.

[0104] Examples of enzymes that can be used in the present invention include oxidoreductases, transferases, hydrolases, lyases, isomerases, and ligases.

[0105] Examples of oxidoreductases include glucose oxidase, alcohol oxidase, glucose dehydrogenase, alcohol dehydrogenase, fructose dehydrogenase, gluconate dehydrogenase, aldehyde dehydrogenase, amine dehydrogenase, succinate dehydrogenase, p-cresol methylhydroxylase, histamine dehydrogenase, fumarate reductase, nitrate reductase, arsenate reductase, sulfite reductase, catalase, peroxidase, and cytochrome P450.

[0106] Examples of transferases include citrate synthase, methyltransferase, phosphotransferase, glycine hydroxymethyltransferase, transketolase, aspartate transaminase, hexokinase, glycerol kinase, creatine kinase, transaminase, and transacylase.

[0107] Examples of hydrolases include carboxylesterase, acetyl-CoA hydrolase, alkaline phosphatase, phospholipase, arylsulfatase, amylase, glucoamylase, cellulase, DNA glycosylase, trypsin, chymotrypsin, pepsin, urease, serine protease, and lipase.

[0108] Examples of lyases include alginate lyase, pyruvate decarboxylase, phosphoketoketolase, citrate lyase, phosphopyruvate hydratase, tryptophan synthase, pectin lyase, aspartate ammonia lyase, cysteine ​​lyase, adenylate cyclase, and ferrochelatase.

[0109] Examples of isomerases include amino acid racemase, tartrate epimerase, glucose-6-phosphate 1-epimerase, maleate isomerase, phenylpyruvate tautomerase, phosphoglucose isomerase, phosphomannomutase, and tyrosine-2,3-aminomutase.

[0110] Examples of synthetic enzymes include tyrosine tRNA ligase, acetyl-CoA synthetase, asparagine synthetase, GMP synthetase, pyruvate carboxylase, and DNA ligase.

[0111] Microorganisms applicable to the present invention include, for example, prokaryotes (bacteria, actinomycetes, archaea), eukaryotes (molds, yeasts, mushrooms, algae, protozoa), etc. Animal and plant cells include, for example, animal cells, plant cells, cultured animal cells, cultured plant cells, etc. Examples of tissues derived from animals and plants include animal tissues and plant tissues.

[0112] Biocatalysts such as enzymes and yeasts are susceptible to the influence of temperature, pH, solvents, or intermolecular electrostatic repulsion, which can easily destroy the three-dimensional molecular structure and reduce their activity, i.e., catalytic ability. Therefore, known methods for long-term storage of biocatalysts include freeze-drying the powder and cryopreservation, in which the biocatalyst is dissolved in a solution at a low concentration and extremely low temperature. However, cryopreservation requires special equipment, and when the frozen solution is thawed for use, the structure of the biocatalyst often changes, reducing its activity. Furthermore, the storage concentration is low, making efficient storage difficult.

[0113] In order to increase the affinity of enzymes, various factors must be considered. For example, it is important to suppress the interaction between enzyme molecules due to the repulsive force (Coulomb interaction) generated by the charge of the enzyme molecules by adding salts, thereby allowing more enzyme molecules to exist per unit volume. It is also important to increase the affinity between the preservation material and the amino acid residues, such as hydroxyl groups, carbonyl groups, and amino groups, which are abundant on the enzyme surface.

[0114] Organic ammonium salts with anions and cations are expected to suppress intermolecular interactions between enzymes and thus increase enzyme affinity, but conventional imidazolium and tetraalkylammonium organic ammonium salts have low affinity for enzyme surfaces. On the other hand, compounds such as polyhydric alcohols (glycerin, propylene glycol, glucose, trehalose, etc.) with hydroxyl groups that have affinity for amino acid residues (hydroxyl, carbonyl, amino, etc.) on the enzyme surface have low inhibitory effects on intermolecular interactions between enzymes and low affinity for enzymes. Furthermore, imidazolim organic ammonium salts with hydroxyl groups have a rigid ring structure and therefore low affinity for enzymes.

[0115] In contrast, the organic ammonium salt used in the biological sample treating agent of the present invention is an organic ammonium salt having a hydrogen-bonding functional group on the cation or on both the cation and the anion, thereby suppressing intermolecular interactions of enzymes, and also having a hydrogen-bonding functional group (hydroxyl group, carboxyl group, ether group, hydrogen bonded to a nitrogen atom) on the cation, which has high affinity with hydroxyl groups, carbonyl groups, amino groups, and other amino residues on the enzyme surface, and its small molecular size and flexible structure result in high affinity.Furthermore, the presence of a hydrogen-bonding functional group on the anion can further increase affinity.

[0116] Maintaining enzyme activity requires maintaining the enzyme's three-dimensional structure. Generally, enzymes possess substrate and reaction specificity, which are expressed through amino acid residues, and function as reaction catalysts. Substrate specificity refers to the ability of an enzyme to recognize and select the structure of a substrate to which it binds, based on the three-dimensional structure of the reactive site and amino acid residues, and to react only with specific substrates. Reaction specificity refers to the ability of an enzyme to catalyze only specific chemical reactions, and is related to the three-dimensional structure of the reactive site, amino acid residues, and metal ions possessed by some enzymes. For example, metal ions present inside enzymes such as oxidoreductases form three-dimensional complexes with amino acid residues to exert catalytic activity. In other words, the loss of substrate specificity, reaction specificity, and metal ions is primarily due to the disruption of the three-dimensional structure of amino acid residues. Therefore, it is important to maintain the enzyme's three-dimensional structure by protecting amino acid residues (e.g., hydroxyl, carbonyl, and amino groups) that contribute to the hydrophilicity of the enzyme surface, as well as hydrophilic amino acid residues (e.g., hydroxyl, carbonyl, and amino groups) and amino acid residues with hydrophobic functional groups inside the enzyme's active site.

[0117] Water and buffers, traditionally used as enzyme solvents, have a high affinity for the hydrophilic sites on the enzyme surface, but they fail to protect the three-dimensional structure of the hydrophobic regions inside the enzyme, which are important for the expression of substrate and reaction specificity, and therefore fail to maintain catalytic activity. Aqueous solutions of the protein bovine serum albumin are sometimes used as stabilizers, but concerns about infectious diseases such as BSE make them difficult to use in the medical field. Aqueous solutions using polyhydric alcohol stabilizers such as glycerin, propylene glycol, glucose, and trehalose maintain the three-dimensional structure of enzymes by integrating the hydrophilic sites on the enzyme surface with the hydroxyl groups of these polyhydric alcohols, and the hydrophilic and hydrophobic alkyl chains of the polyhydric alcohols with the hydrophilic and hydrophobic regions inside the enzyme, where the active sites are located, respectively. However, their preservation stability is limited. Aqueous solutions of surfactants (amino acids) such as glycine and lysine bind to the hydrophobic amino acid residues in the hydrophobic regions inside the enzyme, causing the hydrophobic regions to become hydrophilic due to the generation of electric charges, and migrate to the hydrophilic surface, disrupting the three-dimensional structure of the enzyme and resulting in inactivation. Furthermore, conventionally known organic ammonium salts, such as imidazolium-based and tetraalkylammonium-based organic ammonium salts, which do not have hydrogen-bonding functional groups, cannot protect the hydrophilic amino residues on the surface and inside of the enzyme, and therefore have low affinity for the enzyme.

[0118] In contrast, the organic ammonium salts used in the biological sample treating agent of the present invention protect the enzyme by hydrogen bonding with the hydroxyl, carbonyl, and amino groups of amino acid residues on the surface and inside of the enzyme, via hydrogen bonding in the cations and anions that make up the salt.Furthermore, by simultaneously protecting amino acid residues with internal hydrophobic functional groups with the hydrophobic moiety of the alkyl chain in the organic ammonium salt, the enzyme's three-dimensional structure can be maintained, and catalytic activity can be maintained, even over long periods of time under high-concentration conditions.

[0119] In addition, the organic ammonium salt used in the biological sample processing agent of the present invention consists of a combination of a cation and an anion having a hydrogen-bonding functional group (a hydroxyl group, a carboxyl group, an ether group, or hydrogen bonded to a nitrogen atom), and due to its electrostatic action, it is able to better retain the three-dimensional structure of the enzyme and the activity of the enzyme than nonionic compounds having hydrogen-bonding functional groups.

[0120] Furthermore, the organic ammonium salt used in the biological sample treatment agent of the present invention has a small molecular size and a flexible structure. Therefore, even though it contains a hydroxyl group, the organic ammonium salt penetrates into the interior of a complex three-dimensional structure more efficiently than imidazolium organic ammonium salts, which have a rigid cyclic structure, and protects amino acid residues without three-dimensional distortion, resulting in high retention of enzyme activity.

[0121] Oxidoreductases are enzymes that exert their catalytic action by transferring hydrogen atoms, transferring electrons, and adding oxygen atoms from the substrate. Most of them exert their catalytic action by changing the valence of metal ions in the enzyme due to electron transfer.

[0122] Transferases are enzymes that catalyze the transfer of atomic groups (functional groups) from one substrate to another. Because transfer reactions only occur to functional groups present in the substrate, it is particularly important that the substrate maintain a conformation that allows it to accommodate the reaction. Hydrolases are enzymes that react with the substrate, water, and hydroxyl groups in the amino acid residues of the enzyme to cleave (hydrolyze) specific bonds in the substrate.

[0123] Lysozymes are enzymes that cleave carbon-carbon or carbon-oxygen bonds in substrates without oxidation or hydrolysis. Most of them cleave bonds in substrate molecules by reacting the substrate with a metal ion to generate an intermediate. Isomerases are enzymes that convert substrates into stereoisomers with different spatial arrangements, so the three-dimensional structure of the amino acid residues in the enzyme that bind the substrate is important.

[0124] Synthetic enzymes are enzymes that use the energy of ATP hydrolysis to combine substrates. The reaction occurs via an intermediate formed by the combination of ATP and specific amino acid residues in the enzyme, resulting in the reaction of two substrates to produce the desired product.

[0125] In other words, the amino acid residues, the three-dimensional structure of the amino acid residues, or the metal ions that form three-dimensional complexes with the amino acid residues are important for the activity of any enzyme, and these factors contribute to the expression of the enzyme activity. The biocatalyst solvent of the present invention, due to its structural characteristics, protects the amino acid residues or the metal ions that form complexes, thereby enabling the activity of the biocatalyst to be maintained.

[0126] Furthermore, the biological sample processing agent of the present invention can suppress denaturation caused by heat, among various denaturing factors such as heat (temperature) and pH. Heat breaks hydrogen bonds between amino acid residues, disrupting the enzyme's three-dimensional structure and causing it to denature. However, the biological sample processing agent of the present invention forms a hydrogen-bonding network between the amino acid residues within the enzyme and the hydrogen-bonding functional groups in the organic ammonium salt, thereby more firmly maintaining the three-dimensional structure and suppressing thermal denaturation. In other words, the biological sample processing agent of the present invention can maintain its activity and be stored at a high enzyme concentration for long periods of time, even at room temperature (25°C), which is higher than the typical enzyme storage conditions of -20 to 5°C, and at accelerated temperatures of 40°C, at which enzymes are inactivated.

[0127] Among biological samples, proteins are not particularly limited, but examples of protein types include, in terms of solution properties, acidic proteins that contain a large amount of amino acids with carboxyl groups (aspartic acid, glutamic acid, etc.), alkaline proteins that contain a large amount of amino acids with amino groups (lysine, arginine, histidine, etc.), and neutral proteins that are a balanced combination of these.

[0128] These proteins are classified into two types: simple proteins, which are composed only of amino acids, and complex proteins, which contain other components as well. Simple proteins include albumin, casein, collagen, keratin, protamine, and histones. Complex proteins include glycoproteins (luteinizing hormone, follicle-stimulating hormone, thyroid-stimulating hormone, human chorionic gonadotropin, avidin, cadherins, proteoglycans, mucins, etc.), lipoproteins (chylomicrons, LDL, HDL, etc.), nucleoproteins (histone proteins, telomerase, protamine, etc.), chromoproteins (chlorophyll, etc.), metalloproteins (hemoglobin, cytochrome C, etc.), and phosphoproteins (casein in milk, vitellin in egg yolk, etc.). All enzymes are also proteins.

[0129] Based on their molecular shape, proteins can be classified as fibrous proteins (keratin, collagen, etc.) and globular proteins (hemoglobin, etc.), and based on their function, they can be classified as enzymatic proteins (enzymes), structural proteins (collagen, keratin, etc.), transport proteins (hemoglobin, albumin, apolipoproteins, etc.), storage proteins (ovalbumin, ferritin, hemosiderin in egg white, etc.), contractile proteins (actin, myosin, etc.), defensive proteins (globulins, etc.), and regulatory proteins (calmodulin, etc.).

[0130] Based on molecular and intermolecular structures, examples include those with primary structure (amino acid sequence), secondary structure (α-helix, β-structure, random coil), tertiary structure (specific spatial arrangement), and quaternary structure (hemoglobin, DNA polymerase, ion channels, etc.).

[0131] The molecular weight of the protein is not particularly limited, but proteins of 4,000 to 300,000 are considered.

[0132] Among biological samples, examples of nucleic acids include DNA, RNA, etc. These nucleic acids are known to be easily hydrolyzed in water by their decomposing enzymes, and when water is used as a solvent, they must be dissolved in water from which the decomposing enzymes have been removed during storage. However, by using the biological sample treating agent of the present invention to store nucleic acids and storing them in the form of a nucleic acid-containing solution, nucleic acids can be stored in an environment where nucleases are inactivated, and because the agent is nonvolatile and highly thermally stable, long-term stable storage of nucleic acids is easily possible.

[0133] For the above reasons, when the organic ammonium salt of the present invention is used in a biological sample treatment agent, it is particularly preferred that the organic ammonium salt has a hydrogen-bonding functional group in the anion, i.e., an organic ammonium salt having a hydrogen-bonding functional group in both the cation and the anion. The functional group in the anion includes groups capable of hydrogen bonding, such as oxygen-containing groups, nitrogen-containing groups, sulfur-containing groups, and phosphorus-containing groups, and the above-mentioned carboxylate anions, sulfur-based anions, phosphorus-based anions, cyanide-based anions, and nitrogen oxide-based anions are preferred.

[0134] Preferred examples of the hydrogen-bonding functional group contained in the anion include a hydroxyl group, a carbonyl group, a carboxyl group, a carboxylate group, a sulfonyl group, a sulfate ester group, a phosphate group, and a phosphate ester group, and among these, a hydroxyl group, a carboxyl group, a carboxylate group, a sulfonyl group, and a phosphate group are more preferred.

[0135] As described above, the organic ammonium salt used in the biological sample processing agent of the present invention is solid at 25°C and has hydrogen-bonding functional groups on the cation or both the cation and the anion, resulting in high affinity with biological samples and excellent biological sample preservation properties. When a biological sample is dissolved, dispersed, and stored in an organic ammonium salt that is liquid at 25°C, the solubility of the biological sample and uniform dispersion stability are limited. However, the solid state at 25°C allows for mixing with a biological sample at a higher concentration, enabling storage at a higher concentration. Furthermore, the biological sample processing agent of the present invention allows for long-term storage of biological samples even in high-temperature environments, and for example, allows for long-term storage at high temperatures while maintaining the structures of proteins, nucleic acids, and enzymes, and thus maintaining enzymatic activity.

[0136] The hydrogen-bonding material treating agent of the present invention forms hydrogen bonds with hydrogen-bond-accepting functional groups of biological samples, such as enzymes having carbonyl groups or ether groups, peptides, proteins, nucleic acids, and poorly soluble polysaccharides such as cellulose, and the organic ammonium salt penetrates into the complexly entangled structure of the biological sample, unravelling the biomolecular structure and reducing the interactions between biopolymers. This makes it possible to activate proteins that are in an inactive, denatured state, and is therefore useful as a protein refolding agent.

[0137] Protein refolding refers to restoring a protein that has become insoluble or lost its higher-order structure to its native (activated) higher-order structure. For example, the insoluble or lost-structured protein is solubilized and refolded directly into a refolding solution containing the organic ammonium salt of the present invention, optionally with the addition of a denaturant. Alternatively, the insoluble or lost-structured protein is solubilized with a general protein solubilizer, optionally with the addition of a denaturant, and the solubilized solution is dissolved in a refolding solution containing the organic ammonium salt of the present invention to restore the higher-order structure of the protein, thereby obtaining an active protein.

[0138] The biological sample solution of the present invention comprises the biological sample treating agent of the present invention, a biological sample, and a solvent. The biological sample is preferably a biocatalyst, protein, or nucleic acid. When used as a biological sample solution, the organic ammonium salt of the present invention is highly hydrophilic due to its structural characteristics and has a high affinity for biological samples. Therefore, the organic ammonium salt of the present invention can be used alone in a solid state, or can be mixed with other solvent components such as water or a polar solvent as described above to be used as a solution or dispersion. It can also be used with the addition of additives.

[0139] The biological sample solution includes, for example, a solution containing an activated biological sample and a solution in which the biological sample is preserved while maintaining its active state.

[0140] The storage period of the activated biosample, that is, the biocatalyst, depends on the type of biocatalyst, but can be, for example, 30 days or more, or even 60 days or more.

[0141] Regarding the storage temperature of the biocatalyst, the biocatalyst solution of the present invention can be stored under harsh conditions such as high temperature and humidity, and the biocatalyst can be stored in a liquid state while retaining its activity for a long period of time, for example, at a temperature of 40°C or below. [Example]

[0142] The present invention will be explained in more detail below with reference to examples, but the present invention is not limited to these examples. The compounds of Examples 1 to 108 shown in Tables 1 to 6 were synthesized as follows. Example 1: Synthesis of Compound 1 The compound represented by the following formula was synthesized.

[0143] [ka] Triethanolamine (19.11 g, 0.128 mol) and formic acid (5.89 g, 0.128 mmol) were reacted in 100 mL of water at room temperature for 3 hours, and the water was then distilled off under reduced pressure to give a pale yellow solid. The resulting solid was washed to give Compound 1 (triethanolamine formate) as a white solid. FT-IR (KBr): 3360 cm -1 :OH stretching vibration 2950cm -1 :CH stretching vibration 1558cm -1 COO - stretching vibration 1 H-NMR (D2O 400MHz):δ 3.31(t, 6H, N + C H 2CH2OH), δ 3.85 (t, 6H, N + CH2C H 2OH), δ 8.36 (s, 1H, H COO - ). 13 C-NMR (DO 100MHz): δ 55.4 (N + C H2CH2OH), δ 55.6 (N + CH2 C H2OH), δ 171.0 (H C OO - ).

[0144] <Examples 2 to 87> Compounds 2 to 87 of Examples 2 to 87 shown in Tables 1 to 5 were synthesized by the same synthesis method as in Example 1 and in the blending molar ratios shown in Tables 7 to 9. The physical property values ​​are shown below. Example 2: Synthesis of Compound 2

[0145] [ka] FT-IR (KBr): 3388 cm -1 :OH stretching vibration 2922cm -1 :CH stretching vibration 1543cm -1 COO - stretching vibration 1 H-NMR (D2O 400MHz): δ 0.73-0.77 (m, 3H, NH3 + C(CH2OH)2CH2C H 3), δ 1.50-1.56 (m, 2H, NH3 + C(CH2OH)2C H 2CH3), δ 1.84 (s, 3H, C H 3COO - ), δ 3.49-3.50 (m, 4H, NH3 + C(C H 2OH)2CH2CH3). 13 C-NMR (D2O 100MHz): δ 6.3 (NH3 + C(CH2OH)2CH2 C H3), δ 23.2 (NH3 + C(CH2OH)2 C H2CH3), δ 23.3 ( C H3COO - ), δ 60.5 (NH3 + C (CH2OH)2CH2CH3), δ 63.3 (NH3 + C( C H2OH)2CH2CH3), δ 181.4 (CH3 C OO - ). <Example 3> Synthesis of Compound 3

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[0231] [ka] 2-Amino-2-ethyl-1,3-propanediol (25.00 g, 0.210 mol) and 3-chloro-1,2-propanediol (116.07 g, 1.050 mol) were reacted in 1000 mL of 1-propanol under reflux for 48 hours. The 1-propanol was removed by evaporation under reduced pressure. The resulting liquid was added with THF and washed with heat to obtain a white powder. Sodium hydroxide was added to the resulting white powder and stirred at room temperature for 2 hours. Ethanol was added to precipitate crystals, which were then filtered. The filtrate was removed by evaporation under reduced pressure. The resulting liquid was purified by column chromatography to obtain amine compound 1, listed as Example 88 in Table 10. Amine compound 1 (2.50 g, 0.013 mol) and succinic acid (1.53 g, 0.013 mmol) were reacted in 50 mmL of water at room temperature for 3 hours, and the water was removed by distillation under reduced pressure to give a white solid. The resulting solid was washed to give compound 88 (ammonium succinate) as a white solid. FT-IR (KBr): 3162 m -1:OH stretching vibration 2950cm -1 :CH stretching vibration 1715cm -1 COOH stretching vibration 1578cm -1 COO - Stretching vibration 1 H-NMR (D2O 400MHz): δ 0.86-0.92 (m, 3H, NH2 + C(CH2OH)2CH2C H 3), δ 1.44 (m, 2H, NH2 + C(CH2OH)2C H 2CH3), δ 2.51 (s, 4H, HOOCC H 2C H 2COO - ),δ 2.78-2.96 (m, 2H, NH2 + C H 2CH(OH)), δ 3.61-3.78 (m, 6H, NH2 + C(C H 2OH)2CH2CH 3, NH2 + CH2CH(OH)C H 2OH), δ 3.83-3.88 (m, 1H, NH2 + CH2C H (OH)) 13 C-NMR (D2O 100MHz): δ 9.6 (NH2 + C(CH2OH)2CH2 C H3), δ 22.7 (CH3 C H2CH2), δ 24.6 (NH2 + C(CH2OH)2 C H2CH3), δ 31.4 (HOOC C H2 C H2COO - ),δ 43.6 (NH2 + C (CH2OH)2CH2CH3), δ 45.8 (NH2 + C H2CH(OH)), δ 60.2 (NH2 + C( C H2OH)2CH2CH3), δ 62.8 (NH2+ CH2CH(OH) C H2OH), δ 71.1 (NH2 + CH2 C H(OH) C H2OH), δ 179.7 ( C OOH, C OO - ). Example 89: Synthesis of Compound 89

[0232] [ka] D-glucamine (25.00 g, 0.138 mol) and 3-chloro-1,2-propanediol (76.26 g, 0.690 mol) were reacted in 1000 mL of 1-propanol under reflux for 48 hours, and then the 1-propanol was removed by evaporation under reduced pressure. The resulting liquid was added with THF and washed with heat to obtain a white powder. Sodium hydroxide was added to the resulting white powder, and the mixture was stirred at room temperature for 2 hours. Ethanol was added, and the precipitated crystals were filtered off. The filtrate was removed by evaporation under reduced pressure. The resulting liquid was purified by column chromatography to obtain amine compound 2, which is listed in Example 89 in Table 10. Amine compound 2 (2.50 g, 0.010 mol) and succinic acid (1.16 g, 0.010 mol) were reacted in 50 mL of water at room temperature for 3 hours, and the water was removed by distillation under reduced pressure to give a yellow solid. The resulting solid was washed to give compound 89 (ammonium succinate) as a yellow solid. FT-IR (KBr): 3162 m -1 :OH stretching vibration 2950cm -1 :CH stretching vibration 1715cm -1 :COOH stretching vibration 1578cm -1 COO - stretching vibration 1 H-NMR (D2O 400MHz):δ 2.51 (s, 4H, HOOCC H 2C H 2COO - ), δ 3.00-3.23 (m, 4H, HOCH2(CH(OH))3CH(OH)C H 2NH2+ , NH2 + C H 2CH(OH)), δ 3.51 - 3.74 (m, 7H, HOC H 2(C H (OH))3CH(OH)CH2NH2 + , NH2 + CH2CH(OH)C H 2OH), δ 3.94 - 3.98 (m, 1H, NH2 + CH2C H (OH), δ 4.03 - 4.09 (m, 1H, HOCH2(CH(OH))3C H (OH)CH2NH2 + ). 13 C - NMR (D2O 100MHz):δ 31.4 (HOOC C H2 C H2COO - ), δ 43.2 (HOCH2(CH(OH))3CH(OH) C H2NH2 + ), δ 45.8 (NH2 + C H2CH(OH)), δ 62.8 (NH2 + CH2CH(OH) C H2OH), δ 64.7 (HO C H2(CH(OH))3CH(OH)CH2NH2 + ), δ 70.7 - 72.9 (HOCH2( C H(OH))3 C H(OH)CH2NH3 + , NH2 + CH2 C H(OH) C H2OH), δ 179.7 ( C OOH, C OO - ). Synthesis of Compound 90 in <Example 90>

[0233]

Chemical Structure

[0234] [ka] Amine compound 2 (10.00 g, 0.039 mol) and 3-chloro-1,2-propanediol (21.56 g, 0.195 mol) were reacted in 500 mL of 1-propanol under reflux for 48 hours, after which the 1-propanol was removed by evaporation under reduced pressure. The resulting liquid was added with THF and washed with heat to obtain a white powder. Sodium hydroxide was added to the resulting white powder and stirred at room temperature for 2 hours. Ethanol was added to precipitate crystals, which were then filtered off. The filtrate was removed by evaporation under reduced pressure, and the resulting liquid was purified by column chromatography to obtain amine compound 4, listed in Example 91 in Table 10. Amine compound 4 (2.50 g, 0.009 mol) and oleic acid (2.64 g, 0.009 mol) were reacted in 50 mL of water at room temperature for 3 hours, and the water was removed by distillation under reduced pressure to give a yellow solid. The resulting solid was washed to give compound 91 (ammonium oleate) as a yellow solid. FT-IR (KBr): 3162 m -1 :OH stretching vibration 2950cm -1 :CH stretching vibration 1578cm -1 COO - stretching vibration 1 H-NMR (D2O 400MHz):δ 3.00-3.23 (m, 6H, HOCH2(CH(OH))3CH(OH)C H 2NH + , NH + C H 2CH(OH)), δ 3.51-3.74 (m, 9H, HOCH 2(C H (OH))3CH(OH)CH2NH + , NH + CH2CH(OH)C H 2OH), δ 3.94-3.98 (m, 2H, NH + CH2C H (OH), δ 4.03-4.09 (m, 2H, HOCH2(CH(OH))3C H (OH)CH2NH + ).δ 0.89 (t, 3H, C H 3CH2), δ 1.27 (m, 20H, CH3(C H 2)6CH2, (C H 2)4CH2CH2COO - ),δ 1.53 (m, 2H, C H 2CH2COO - ), δ 2.00 (m, 4H, C H 2CH=CHC H 2), δ 2.14 (t, 2H, CH2C H 2COO - ),δ 5.32 (m, 2H, C H =C H ). 13 C-NMR (D2O 100MHz):δ 43.2 (HOCH2(CH(OH))3CH(OH) C H2NH + ), δ 45.8 (NH + C H2CH(OH)), δ 62.8 (NH + CH2CH(OH) C H2OH), δ 64.7 (HO C H2(CH(OH))3CH(OH)CH2NH + ), δ 70.7-72.9 (HOCH2( C H(OH))3 C H(OH)CH2NH + , NH + CH2 C H(OH) C H2OH).δ 14.0 ( CH3CH2), δ 22.6 (CH3 C H2), δ 26.4 ( C H2CH2COO - ), δ 27.2 ( C H2CH=CH C H2), δ 29.3 (CH3CH2CH2( C H2)4, ( C H2)4CH2CH2COO - ), δ 31.9 (CH3CH2 C H2), δ 37.8 ( C H2CH2COO - ), δ 129.7 ( C H= C H), δ 181.7 ( C OO - ). Example 92: Synthesis of Compound 92

[0235] [ka] It was synthesized by the same synthesis method as in Example 90 and in the blending molar ratios shown in Table 10. The physical properties are shown below. FT-IR (KBr): 3162 m -1 :OH stretching vibration 2950cm -1 :CH stretching vibration 1715cm -1 :COOH stretching vibration 1578cm -1 COO - stretching vibration 1 H-NMR (D2O 400MHz):δ 2.51 (s, 4H, HOOCC H 2C H 2COO - ), δ 3.00-3.23 (m, 4H, HOCH2(CH(OH))3CH(OH)C H 2NH2 + , NH2 + C H 2CH(OH)), δ 3.51-3.74 (m, 7H, HOC H 2(C H (OH))3CH(OH)CH2NH2 + ,NH2 + CH2CH(OH)C H 2OH), δ 3.94-3.98 (m, 1H, NH2 + CH2C H (OH), δ 4.03-4.09 (m, 1H, HOCH2(CH(OH))3C H (OH)CH2NH2 + ).δ 2.51 (s, 4H, HOOCC H 2C H 2COO - ). 13 C-NMR (D2O 100MHz):δ 31.4 (HOOC C H2 C H2COO - ), δ 43.2 (HOCH2(CH(OH))3CH(OH) C H2NH2 + ), δ 45.8 (NH2 + C H2CH(OH)), δ 62.8 (NH2 + CH2CH(OH) C H2OH), δ 64.7 (HO C H2(CH(OH))3CH(OH)CH2NH2 + ), δ 70.7-72.9 (HOCH2( C H(OH)3 C H(OH)CH2NH3 + , NH2 + CH2 C H(OH) C H2OH). δ 31.4 (HOOC C H2 C H2COO - ), δ 179.7 ( C OOH, C OO - ). Example 93: Synthesis of Compound 93

[0236] [ka] It was synthesized by the same synthesis method as in Example 91 and in the blending molar ratios shown in Table 10. The physical properties are shown below. FT-IR (KBr): 3162m -1 :OH stretching vibration 2950cm -1 :CH stretching vibration 1715cm -1 COOH stretching vibration 1578cm -1 COO - Stretching vibration 1 H-NMR (D2O 400MHz): δ 3.00-3.23 (m, 6H, HOCH2(CH(OH))3CH(OH)C H 2NH + NH + C H 2CH(OH)), δ 3.51-3.74 (m, 9H, HOC H 2(C H (OH))3CH(OH)CH2NH + , NH + CH2CH(OH)C H 2OH), δ 3.94-3.98 (m, 2H, NH + CH2C H (OH), δ 4.03-4.09 (m, 2H, HOCH2(CH(OH))3C H (OH)CH2NH + ).δ 2.51 (s, 4H, HOOCC H 2C H 2COO - ). 13 C-NMR (D2O 100MHz): δ 43.2 (HOCH2(CH(OH))3CH(OH) C H2NH + ), δ 45.8 (NH + C H₂CH(OH)), δ 62.8 (NH₃) + CH2CH(OH) C H2OH), δ 64.7 (HO C H2(CH(OH))3CH(OH)CH2NH + ), δ 70.7-72.9 (HOCH2( C H(OH))3 C H(OH)CH2NH + NH + CH2C H(OH) C H2OH). δ 31.4 (HOOC C H2 C H2COO - ), δ 179.7 ( C OOH, C OO - ). Example 94: Synthesis of Compound 94

[0237] [ka] Amine compound 3 (10.00 g, 0.037 mol) and 3-chloro-1,2-propanediol (40.90 g, 0.370 mol) were reacted in 200 mL of acetonitrile at 130°C under pressure for 4 hours, and the acetonitrile was removed by distillation under reduced pressure. The resulting solid was added with THF and washed with heat to obtain a pale yellow powder. Water was added to this pale yellow powder, and the water was passed through an anion exchange resin to obtain amine compound 5, which is listed as Example 94 in Table 10. Amine compound 5 (5.00 g, 0.014 mol) and isostearic acid (3.98 g, 0.0140 mol) were reacted in 50 mL of water at room temperature for 3 hours, and the water was removed by distillation under reduced pressure to give a yellow solid. The resulting solid was washed to give compound 94 (quaternary ammonium isostearate) as a yellow solid. FT-IR (KBr): 3350 cm -1 :OH stretching vibration 2940cm -1 :CH stretching vibration 1560cm -1 COO - stretching vibration 1 H-NMR (D2O 400MHz): δ 0.86-0.92 (m, 6H, N + C(CH2OH)2CH2C H 3. C H 3CH2CH2), δ 1.44 (m, 2H, N + C(CH2OH)2C H 2CH3), δ 2.78-2.96 (m, 6H, N + C H2CH(OH)), δ 3.61-3.78 (m, 10H, N + C(C H 2OH)2CH2CH 3, N + CH2CH(OH)C H 2OH), δ 3.83-3.88 (m, 3H, N + CH2C H (OH)). δ 0.83-0.90 (m, 6H, C H 3(CH2)8CH((CH2)6C H 3)COO - ), δ1.08-1.58 (m, 28H, CH3(C H 2)8CH((C H 2)6CH3)COO - ), δ 2.24-2.28 (m, 1H, CH3(CH2)8C H ((CH2)6CH3)COO - ). 13 C-NMR (D2O 100MHz): δ 9.6 (N + C(CH2OH)2CH2 C H3), δ 24.6 (N + C(CH2OH)2 C H2CH3), δ 43.6 (N + C (CH2OH)2CH2CH3), δ 45.8 (N + C H2CH(OH)), δ 60.2 (N + C( C H2OH)2CH2CH3), δ 62.8 (N + CH2CH(OH) C H2OH), δ 71.1 (N + CH2 C H(OH) C H2OH). d 14.1 ( C H3CH2), δ 22.7 (CH3 C H--2CH2), δ 27.1 ( C H2CH2CHCOO - ), δ 30.0 (CH3CH2CH2( C H2)4CH2CH2CH(CH2CH2(C H2)2CH2CH2CH3)COO - ,CH2 C H2CHCOO - ), δ 31.9 (CH3CH2 C H2), δ 37.2 ( C HCOO - ), δ 182.1 (CH C OO - ). Example 95: Synthesis of Compound 95

[0238] [ka] Amine compound 4 (10.00 g, 0.030 mol) and 3-chloro-1,2-propanediol (33.16 g, 0.300 mol) were reacted in 200 mL of acetonitrile at 130°C under pressure for 4 hours, and the acetonitrile was removed by distillation under reduced pressure. The resulting solid was added with THF and washed with heat to obtain a pale yellow powder. Water was added to this pale yellow powder, and the water was passed through an anion exchange resin to obtain amine compound 6, which is listed as Example 95 in Table 10. Amine compound 6 (5.00 g, 0.012 mol) and succinic acid (1.42 g, 0.012 mol) were reacted in 50 mL of water at room temperature for 3 hours, and the water was removed by distillation under reduced pressure to give a yellow solid. The resulting solid was washed to give compound 95 (quaternary ammonium succinate) as a yellow solid. FT-IR (KBr): 3344 cm -1 :OH stretching vibration 2920cm -1 :CH stretching vibration 1715cm -1 :COOH stretching vibration 1554cm -1 COO - stretching vibration 1 H-NMR (D2O 400MHz): δ 3.00-3.23 (m, 8H, HOCH2(CH(OH))3CH(OH)C H 2N + , N + C H 2CH(OH)), δ 3.51-3.74 (m, 11H, HOC H2(C H (OH))3CH(OH)CH2N + , N + CH2CH(OH)C H 2OH), δ 3.94 - 3.98 (m, 3H, N + CH2C H (OH)), δ 4.03 - 4.09 (m 1H, HOCH2(CH(OH))3C H (OH)CH2N + ).δ 2.51 (s, 4H, HOOCC H 2C H 2COO - ). 13 C-NMR (D2O 100MHz): δ 43.2 (HOCH2(CH(OH))3CH(OH) C H2N + ), δ 45.8 (N + C H2CH(OH)), δ 62.8 (N + CH2CH(OH) C H2OH), δ 64.7 (HO C H2(CH(OH))3CH(OH)CH2N + ),δ 70.7 - 72.9 (HOCH2( C H(OH))3 C H(OH)CH2N + , N + CH2 C H(OH)CH2OH). δ 31.4 (HOOC C H2 C H2COO - ), δ 179.7 ( C OOH, C OO - ).

[0239] <Examples 96 - 108> Compounds 96 to 101, 103, 105, and 107 of Examples 96 to 101, 103, 105, and 107 shown in Table 6 were synthesized using the same synthesis method as in Example 94 and the blending molar ratios shown in Table 10. Furthermore, compounds 102, 104, 106, and 108 of Examples 102, 104, 106, and 108 were synthesized using the same synthesis method as in Example 95 and the blending molar ratios shown in Table 10. Physical property values ​​are shown below. Example 96: Synthesis of Compound 96

[0240] [ka] FT-IR (KBr): 3398 cm -1 :OH stretching vibration 2922cm -1 :CH stretching vibration 1715cm -1 :COOH stretching vibration 1560cm -1 COO - stretching vibration 1 H-NMR (D2O 400MHz): δ 0.86-0.92 (m, 6H, N + C(CH2OH)2CH2C H 3. C H 3CH2CH2), δ 1.44 (m, 2H, N + C(CH2OH)2C H 2CH3), δ 2.78-2.96 (m, 6H, N + C H 2CH(OH)), δ 3.61-3.78 (m, 10H, N + C(C H 2OH)2CH2CH 3, N + CH2CH(OH)C H 2OH), δ 3.83-3.88 (m, 3H, N + CH2C H (OH)). δ 2.51 (s, 4H, HOOCC H 2C H 2COO - ). 13 C-NMR (DO 100MHz): δ 9.6 (N +C(CH2OH)2CH2 C H3), δ 24.6 (N + C(CH2OH)2 C H2CH3), δ 43.6 (N + C (CH2OH)2CH2CH3), δ 45.8 (N + C H₂CH(OH)), δ 60.2 (N + C( C H2OH)2CH2CH3), δ 62.8 (N + CH2CH(OH) C H2OH), δ 71.1 (N + CH2 C H(OH) C H2OH). δ 31.4 (HOOC C H2 C H2COO - ), δ 179.7 ( C OOH, C OO - ). <Example 97> Synthesis of Compound 97

[0241]

change

[0242] [Chemical Structure] FT-IR (KBr): 3398 cm -1 : O-H stretching vibration 2922 cm -1 : C-H stretching vibration 1560 cm -1 : COO - stretching vibration 1 H-NMR (D2O 400 MHz): δ 0.86 - 0.92 (m, 6H, N + C(CH2OH)2CH2C H 3, C H 3CH2CH2), δ 1.44 (m, 2H, N + C(CH2OH)2C H 2CH3), δ 2.78 - 2.96 (m, 6H, N + C H 2CH(OH)), δ 3.61 - 3.78 (m, 10H, N + C(C H 2OH)2CH2CH 3, N + CH2CH(OH)C H 2OH), δ 3.83 - 3.88 (m, 3H, N + CH2C H (OH)). δ 6.80 - 6.86 (m, 2H, C(OH)C H CH, C(COO - )CHC H CH), δ 7.35 - 7.39 (m, 1H, C H CHC(OH)), δ 7.68 - 7.73 (m, 1H, C(COOH)C H CH). 13 C-NMR (D2O 100 MHz): δ 9.6 (N + C(CH2OH)2CH2 C H3), δ 24.6 (N + C(CH2OH)2 C H2CH3), δ 43.6 (N + C (CH2OH)2CH2CH3), δ 45.8 (N + C H2CH(OH)), δ 60.2 (N+ C( C H2OH)2CH2CH3), δ 62.8 (N + CH2CH(OH) C H2OH), δ 71.1 (N + CH2 C H(OH) C H₂OH). δ 116.2 (C(OH)₂) C HCH), δ 118.0 (CH C (COO - )C(OH)), δ 119.3 (C(COO - CH C HCH), δ 130.5 (C(COO - ) C HCH), δ 133.9 ( C HCHC(OH)), δ 159.6 (C C (OH)C), δ 175.5 (C C OO - ). <Example 99> Synthesis of Compound 99

[0243]

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[0244]

Chem.

[0245]

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[0246]

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[0247]

Chemical Structure

[0248]

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[0249]

change

[0250]

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[0251]

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[0252]

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[0253] [ka] Compound 109 of Example 109 shown in Table 6 was synthesized using the same synthesis method as in Example 1 and the blending molar ratio shown in Table 10. The physical property values ​​are shown below. FT-IR (KBr): 3145 cm -1 :OH stretching vibration 2946cm -1 :CH stretching vibration 1572cm -1 COO - stretching vibration 1 H-NMR (D2O 400MHz):δ 2.28 (s, 4H, - OOCC H 2C H 2COO - ), δ 3.61 (s, 12H, NH3 + C(C H 2OH)3). 13 C-NMR (DO 100MHz): δ 34.0( - OOC C H2 C H2COO - ), δ 59.4 (NH3 + C (CH2OH)3), δ 61.3 (NH3 + C( C H2OH)3), δ 182.3 ( COO - ).

[0254] [Table 1]

[0255] [Table 2]

[0256] [Table 3]

[0257] [Table 4]

[0258] [Table 5]

[0259] [Table 6]

[0260] <Comparative Example 1> Compound 110 Tributylammonium Lactate It was synthesized by the method described in JP 2014-131974 A.

[0261] <Comparative Example 2> Compound 111 Choline acetate It was synthesized using choline hydroxide and acetic acid with reference to the method described in JP 2014-131974 A.

[0262] <Comparative Example 3> Compound 112 1-Butyl-3-methylimidazolium bromide The compound was synthesized using 1-butyl-3-methylimidazolium tetrafluoroborate, an ion exchange resin, and hydrobromic acid, with reference to the method described in JP 2016-041682 A.

[0263] <Comparative Example 4> Compound 113 As D(+)-glucose, a reagent manufactured by Kanto Chemical Co., Ltd. was used.

[0264] <Comparative Example 5> Compound 114 Nippipeptide FCP-AS-L (manufactured by Nippi Corporation) was used as gelatin.

[0265] <Comparative Example 6> Compound 115 Albumin used was that manufactured by Nacalai Tesque Inc. (bovine-derived, general grade, pH 5.2).

[0266] <Comparative Example 7> Compound 116 As the urease, a reagent (derived from jack bean) manufactured by Wako Pure Chemical Industries, Ltd. was used.

[0267] <Comparative Example 8> Compound 117 As a buffer for urease, a 10 mM aqueous solution of potassium dihydrogen phosphate adjusted to pH 7.5 with sodium hydroxide was used.

[0268] <Comparative Example 9> Compound 118 Cytochrome C used was a reagent manufactured by Nacalai Tesques Inc. (Horse Heart, molecular weight 12384).

[0269] <Comparative Example 10> Compound 119 As a buffer for cytochrome C and DNA, a 50 mM phosphate buffer at pH 7.4 prepared from 50 mM potassium dihydrogen phosphate and 50 mM potassium dihydrogen phosphate was used.

[0270] <Comparative Example 11> Compound 120 DNA prepared by the method described in JP 2014-131974 A was used. <Comparative Example 12> Compound 121 Tetra-n-butylammonium bromide used was a reagent manufactured by Kanto Chemical Co., Ltd.

[0271] <Reference example 1> Compound 122 It was synthesized by the method described in JP 2014-131974 A.

[0272] <Reference Example 2> Compound 123 It was synthesized by the method described in JP 2014-131974 A.

[0273] The compounds of the above Examples and Comparative Examples were subjected to the following measurements and evaluations. 1. Properties at room temperature (25°C) The compounds of Examples 1 to 109 were added to a screw tube and dried under reduced pressure to form an anhydrous product, and their properties (liquid, solid) at room temperature (25°C) were confirmed. The results are shown in Tables 7 to 10. In Tables 7 to 10, "solid" indicates that the compound is solid at room temperature (25°C). As a result of the reduced pressure drying treatment, all of the compounds of Examples 1 to 109 were solid at room temperature (25°C).

[0274] Therefore, compared with conventional organic ammonium salts, the organic ammonium salt of the present invention has a polyhydroxyalkyl group, which results in a stronger cationic structure with stronger hydrogen bonds within the molecule, making it easier to crystallize. Therefore, it has been found that the melting point can be influenced by the selection of functional groups and characteristic groups in the structural design, and a wide range of various anions can be applied.

[0275] [Table 7]

[0276] [Table 8]

[0277] [Table 9]

[0278] [Table 10]

[0279] 2.Enzyme long-term stability (1) A 50% aqueous solution of the compound was prepared using the compound listed in Table 11 as a stabilizer, and urease was added and dissolved to achieve the enzyme concentration listed in Table 11. The water was then distilled off under reduced pressure. After distillation under reduced pressure, the resulting solid sample was placed in a thermo-hygrostat set at 40°C and 80% RH, temperatures and humidity levels higher than those typically used for storing enzymes. After leaving the sample for a predetermined period, each sample was collected and the activity retention of the enzyme stored in each compound was measured using the method described below, thereby confirming the retention of the three-dimensional structure of the enzyme and the stabilizing effect of each compound.

[0280] <Hydrolytic enzyme: urease activity measurement> The activity of urease was measured by quantifying the ammonium ions produced by the decomposition of urea by the enzymatic reaction of urease using the indophenol method.

[0281] First, 100 mL of 1 mM substrate solution (prepared by dissolving urea as a substrate in 10 mM phosphate buffer at pH 7.5) was placed in an Erlenmeyer flask and preheated at 30° C. for about 30 minutes.

[0282] Next, the sample left for a predetermined period at the set concentration and temperature shown in Table 11 was added to the above substrate solution so that the amount of enzyme was 0.5 mg, and the reaction was carried out at 30°C for 60 minutes.

[0283] After the reaction, 0.1 mL of the reaction solution was sampled, and immediately 2 mL of a phenol solution (prepared by dissolving 10 g of phenol and 50 mg of sodium pentacyanonitrosylferrate (III) in ion-exchanged water, and then adding ion-exchanged water to make 1000 mL) and 2 mL of a sodium hypochlorite solution (prepared by dissolving 5 g of sodium hydroxide and 8.4 mL of a 5% sodium hypochlorite solution in ion-exchanged water, and then adding ion-exchanged water to make 1000 mL) were added, followed by reaction in a constant temperature bath at 37°C for 20 minutes.

[0284] The absorbance of this reaction solution at a wavelength of 635 nm was measured using a UV-visible spectrophotometer (V-550: JASCO Corporation), and the amount of ammonium ion produced was calculated from the amount of indophenol obtained, to calculate the urease activity. Ammonium ion was quantified using a calibration curve obtained by preparing ammonium ion solutions in the concentration range of 0.1 to 3.0 mM and quantifying them using the indophenol method in the same manner as above.

[0285] The enzyme activity value, which served as the basis for the enzyme activity retention rate, was calculated as follows: Urease powder stored at an appropriate temperature was dissolved in a buffer (10 mM phosphate buffer, pH 7.5) to prepare an enzyme solution with an enzyme concentration of 50 mg / mL. Immediately after preparation, this solution was added to the substrate solution so that the amount of enzyme was 0.5 mg, as described above, and the enzyme reaction was carried out. The enzyme activity retention rate was calculated based on the amount of ammonium ions quantified by the indophenol method.

[0286] In the results for the 40°C condition in Table 11, the activity retention rate of the imidazolim-based organic ammonium salt, the enzyme stored in a common additive, and the enzyme stored without any additives decreased to 0 to 3% after 30 days, whereas the compound of the present invention showed a higher activity retention rate than the comparative example after 30 days, regardless of the stabilizer:enzyme ratio.

[0287] Furthermore, in Reference Examples 1 and 2, which are liquid at 25°C, the enzyme could not be dissolved at a stabilizer:enzyme ratio of 1:1 or 0.1:1, making storage difficult. However, the compound of the present invention is a mixture of solids, making it possible to store the enzyme at a high concentration. In other words, it was suggested that the compound of the present invention retains the enzyme activity and the three-dimensional structure of the enzyme at a high concentration, at a high temperature, and for a long period of time.

[0288] [Table 11] A 50% aqueous solution of the compounds in Table 12 was prepared, and urease was added and dissolved to achieve the enzyme concentration shown in Table 12. The solution was then left in a thermo-hygrostat set at 40°C and 80% RH, which are higher than the temperature and humidity at which enzymes are generally stored. After leaving the solution for a predetermined period, each sample was collected, and the activity retention rate of the enzyme stored in each compound was measured using the method described below, thereby confirming the retention of the three-dimensional structure of the enzyme and the stabilizing effect of each compound.

[0289] In the results of the aqueous solution stored at 40°C in Table 12, the activity retention rate of the enzyme stored in imidazolim-based organic ammonium salt, 50% aqueous solution of a general additive, and buffer decreased to 0 to 2% after 30 days, whereas the 50% aqueous solution containing the compound of the present invention showed a higher activity retention rate than the comparative example after 30 days.

[0290] In other words, it was suggested that the compound of the present invention retains the enzyme activity not only in the solid state but also in an aqueous solution under conditions of high concentration, high temperature, and long term, and has a high degree of retention of the enzyme's three-dimensional structure.

[0291] [Table 12]

[0292] 3. Long-term protein stability A 50% aqueous solution of each compound in Table 13 was prepared, and cytochrome C was added and dissolved to achieve the protein concentration shown in Table 13. The water was then removed by distillation under reduced pressure. After distillation under reduced pressure, the resulting solid sample was placed in a thermo-hygrostat set at 40°C and 80% RH, which are temperatures and humidities typically higher than those used for storing proteins. After leaving the sample for a predetermined period, each sample was collected, and the IR and UV spectra of the protein stored in each compound were measured using the following method to confirm the long-term stability of the protein.

[0293] First, we confirmed the changes in amide absorption in IR spectra in detail and confirmed the higher-order structure of the protein (turn, α-helix, random coil, β-sheet). -1 ), amide II region (1500-1600 cm -1 ) was measured using the ATR method with a Fourier transform infrared spectrophotometer (FT / IR-6100: JASCO Corporation), and peaks were detected from the difference between an organic ammonium salt sample without added cytochrome C (blank) and an organic ammonium salt sample with added cytochrome C (sample).

[0294] In addition, the active state of cytochrome C (Fe) can be determined by the absorption of UV spectrum. 2+ : Reduced type, Fe 3+ The organic ammonium salt sample containing added cytochrome C was diluted to 1% with 50 mM phosphate buffer (prepared with 50 mM potassium dihydrogen phosphate and 50 mM potassium dihydrogen phosphate) at pH 7.4, and immediately thereafter measured in a quartz cell with a 2 mm optical path width on a UV-visible spectrophotometer. The results are shown in Table 13. Products showing long-term stability to proteins were marked with a circle, and products not showing long-term stability were marked with an x.

[0295] In identifying the amide absorption by IR spectroscopy, the higher-order structure of the protein was confirmed by comparing it with literature values ​​(Chem.Commun 2005, 4804-4806, Biomacromolecules 2010, 11, 2944-2948, Protein Science Society Archives, 2, e054 (2009)). IR measurement of the cytochrome C powder before addition revealed a peak at 1645 cm in the amide I region. -1 and 1537 cm in the amide II region -1 Next, cytochrome C in phosphate buffer did not denature (amide I region: 1653 cm). -1 , amide II region: 1547 cm -1 ), which confirmed that it has an α-helix structure. The cytochrome C in the organic ammonium salt of the present example showed the same measurement results as in the phosphate buffer (amide I region: 1652-1655 cm).-1 , amide II region: 1545-1549 cm -1 ), which confirmed that the cytochrome C stored in the organic ammonium salts of the Examples did not undergo a denatured structure but maintained its α-helical structure. In other words, dissolving a solid of an inactive denatured protein (cytochrome C) using the organic ammonium salt of the present invention resulted in the release of protein activity, demonstrating the refolding effect and maintaining cytochrome C without denaturation.

[0296] Cytochrome C transports Fe during intracellular electron transport. 2+ (reduced form) and Fe 3+ The reduced form reversibly changes its state to an oxidized form (a reduced form), maintaining its secondary structure in the active state. In the UV spectrum, the reduced form exhibits peaks at 550 nm in the α band, 521 nm in the β band, and 415 nm in the γ band. The oxidized form exhibits no clear peaks in the α band and β band, and the γ band shifts to a lower wavelength of approximately 396 nm. In the inactive state, the cytochrome C denatured, and the peaks in the α band, β band, and γ band disappeared. Compared with the peaks in the comparative example, the example exhibited reduced-type peaks in the α band, β band, and γ band. This confirmed that the cytochrome C stored in the organic ammonium salt of the present invention maintained its secondary structure in the active, reduced state.

[0297] That is, the product of the present invention is an excellent protein preservation material that does not denature proteins even under high temperature conditions for a long period of time, and furthermore, it has been shown to be useful as a protein refolding agent.

[0298] [Table 13] Furthermore, the results of the aqueous solution at 40°C in Table 14 confirmed that the refolding effect and retention of secondary structure were observed in the 50% aqueous solution containing the compound of the present invention, similar to storage in a solid sample.

[0299] In other words, the product of the present invention is an excellent protein preservation material that does not denature proteins even when used as an aqueous solution, even under high-temperature conditions, for a long period of time, and is also shown to be useful as a protein refolding agent.

[0300] [Table 14]

[0301] 4.DNA long-term stability

[0302] A 50% aqueous solution of the compounds in Table 15 was prepared, and DNA was added to the solution to the concentration shown in Table 15 and dissolved. The water was then removed by evaporation under reduced pressure. After evaporation under reduced pressure, the resulting solid sample was placed in a thermo-hygrostat set at 40°C and 80% RH, temperatures and humidity levels higher than those typically used for storing DNA. After leaving the sample for a predetermined period, each sample was collected, and the UV spectrum of the DNA stored in each compound was measured using the method described below to confirm the long-term stability of the DNA.

[0303] The results are shown in Table 15. Products that exhibited long-term stability to DNA were marked with a circle, and products that did not have long-term stability were marked with an x.

[0304] In an active state, DNA maintains a double helix structure and has a peak in the UV spectrum absorption at around 260 nm. However, when denatured, the relative absorbance of DNA UV absorption increases significantly. For all of the example compounds shown in Table 15, the difference between a 0.1 wt% DNA solution and a sample (blank) containing no dissolved DNA showed a peak at 258 nm representing the absorption of DNA in organic ammonium salts, which was similar to the absorption (259 nm) of DNA dissolved in water (1 wt%). This confirmed that DNA dissolved in organic ammonium salts maintained an active double helix structure.

[0305] That is, it was demonstrated that the product of the present invention has excellent storage stability for DNA and is useful as a storage material for nucleic acids such as DNA.

[0306] [Table 15]

[0307] Furthermore, the results of the aqueous solution at 40°C in Table 16 confirmed that DNA retains its active double helix structure even in a 50% aqueous solution containing the compound of the present invention, similar to storage in a solid sample.

[0308] That is, it was demonstrated that the product of the present invention has excellent storage stability for DNA even when used as an aqueous solution, and is useful as a storage material for nucleic acids such as DNA.

[0309] [Table 16]

[0310] 5.Enzyme long-term stability (2) To prepare samples by distilling off the solvent from the dissolution solution in Table 17, a 50% aqueous solution of the compound was prepared, urease was added and dissolved to give the enzyme concentration shown in Table 17, and then the water was distilled off under reduced pressure to obtain a solid sample. On the other hand, to mix in a mortar, the product of the present invention and urease having the enzyme concentration shown in Table 17 were mixed in a mortar to obtain a solid sample.

[0311] Each sample was then placed in a thermo-hygrostat set at 40°C and 80% RH, which are higher than the temperature and humidity typically used for storing enzymes. After leaving the samples for a predetermined period, the activity retention of the enzyme stored in each compound was measured using the following method, and the retention of the three-dimensional structure and stabilization effect of each compound on the enzyme were confirmed.

[0312] The solid sample prepared from the dissolution solution showed a higher activity retention rate than the sample mixed in a mortar.These results demonstrate that by dissolving the enzyme in the product of the present invention, it is possible to maintain the three-dimensional structure inside the enzyme, and to obtain a biological sample treatment agent that exhibits high activity retention.

[0313] [Table 17]

[0314] 6. Metal oxide dispersibility test For each compound of each Example and Comparative Example shown in Table 18, 0.25 g of each compound, 0.50 g of water, and 0.10 g of zirconium (IV) oxide (Wako Pure Chemical Industries, Ltd.) were mixed in a planetary centrifugal mixer (Thinky Corporation, ARE-310) at 2000 rpm for 1 minute 5 times, and the dispersion state was then visually inspected. Those in which the zirconium oxide was dispersed were evaluated as ○, and those in which the zirconium oxide was not dispersed and had settled were evaluated as ×. The results are shown in Table 18.

[0315] In all of the compounds of the Examples, zirconium oxide was dispersed well, and a dispersion was obtained, whereas in the compound of Comparative Example 12, zirconium oxide immediately precipitated and did not disperse.

[0316] In other words, the organic ammonium salt of the present invention takes advantage of the structural feature of being composed of a quaternary ammonium cation having many hydrogen bond-donating and coordinating hydroxyl groups, thereby increasing the affinity with the hydrogen bond-accepting oxygen atoms of zirconium oxide, and furthermore, it is believed that the affinity between the hydroxyl groups of the quaternary ammonium salt and the coordinating metal atoms of zirconium oxide results in good dispersion of zirconium oxide.

[0317] [Table 18] For the compounds of each Example and Comparative Example shown in Table 19, 0.25 g of each compound, 0.50 g of water, and 0.10 g of titanium(IV) oxide (Wako Pure Chemical Industries, Ltd.) were mixed in a planetary centrifugal mixer (Thinky Corporation, ARE-310) at 2000 rpm for 1 minute 5 times, and the dispersion state was then visually confirmed. Those in which titanium oxide was dispersed were evaluated as ○, and those in which it was not dispersed and had settled were evaluated as ×. The results are shown in Table 19.

[0318] All of the compounds of the Examples dispersed titanium oxide well, and dispersions were obtained, whereas the compound of Comparative Example 12 had poor dispersibility, with titanium oxide settling.

[0319] In other words, the organic ammonium salt of the present invention takes advantage of the structural feature of being composed of a quaternary ammonium cation having many hydrogen bond-donating and coordinating hydroxyl groups, thereby increasing the affinity with the hydrogen bond-accepting oxygen atoms of titanium oxide, and furthermore, it is believed that the affinity between the hydroxyl groups of the quaternary ammonium salt and the coordinating metal atoms of titanium oxide results in good dispersion of titanium oxide.

[0320] These results suggest that the organic ammonium salts of the present invention have excellent affinity with inorganic materials such as metals and metal oxides having hydrogen bond-accepting functional groups, and are useful as treatment agents for these inorganic materials, cosmetics, etc.

[0321] [Table 19]

[0322] 7. Carbon nanotube dispersion test For the compounds of each Example and Comparative Example shown in Table 20, 0.25 g of each compound, 0.75 g of water, and 0.025 g of carbon nanotubes (multi-walled, 3 to 20 nm) (Wako Pure Chemical Industries, Ltd.) were mixed in a planetary centrifugal mixer (Thinky Corporation, ARE-310) at 2000 rpm for 1 minute 5 times, and the dispersion state was then visually confirmed. Those in which the carbon nanotubes were dispersed were evaluated as ○, and those in which the carbon nanotubes had not dispersed and had settled were evaluated as ×. The results are shown in Table 20.

[0323] As a result, all of the compounds of the examples dispersed the carbon nanotubes well, and a low-viscosity dispersion liquid that was easy to handle was obtained, whereas the carbon nanotubes in the compound of Comparative Example 12 settled and did not disperse.

[0324] In other words, the organic ammonium salt of the present invention takes advantage of the structural feature of being composed of a quaternary ammonium cation and a cation having many hydrogen bond-donating hydroxyl groups, thereby increasing its affinity with the hydrogen bond-accepting carbon-carbon unsaturated bonds (π electron system) of carbon nanotubes, and is thought to have resulted in good dispersion of the carbon nanotubes.

[0325] These results suggest that the organic ammonium salt of the present invention has excellent affinity with compounds and materials having carbon-carbon unsaturated bonds and is useful as a treatment agent for such materials.

[0326] [Table 20]

[0327] 8. Organic dye dispersibility test For each compound of each Example and Comparative Example shown in Table 21, 0.2 g of each compound, 0.7 g of water, and 0.1 g of the organic dye Varifast Black 3830 (manufactured by Orient Chemical Industry Co., Ltd.) were ultrasonically dispersed, left to stand for 24 hours, and the dispersion state 24 hours after mixing was visually confirmed. Those in which the organic dye was dispersed were evaluated as ○, and those in which particles had visually settled were evaluated as ×. The results are shown in Table 21. As a result, all of the compounds of the examples dispersed the organic dye well, and a uniform dispersion was obtained, whereas the organic dye in the compound of Comparative Example 12 precipitated and did not disperse.

[0328] In other words, the organic ammonium salt of the present invention takes advantage of the structural feature of being composed of a quaternary ammonium cation having many hydrogen bond-donating and coordinating hydroxyl groups, thereby increasing the affinity with the nitrogen and oxygen atoms of the hydrogen bond-accepting organic dye, and further increasing the affinity between the hydroxyl groups of the quaternary ammonium salt and the metal atom (chromium) of the coordinating organic dye, which is thought to result in good dispersion.

[0329] These results suggest that the organic ammonium salts of the present invention are useful as processing agents for organic compounds and materials having a hydrogen bond-accepting functional group, including organic dyes.

[0330] [Table 21]

[0331] 9.Redispersibility Dispersions of zirconium oxide, titanium oxide, and the organic dye Varifast Black 3830 of Examples 28, 36, 48, and 109 and Comparative Example 12 shown in Table 22 were dehydrated under reduced pressure at 50°C for 3 hours to obtain dried compounds. 0.50 g of water was added to the zirconium oxide and titanium oxide samples, and 0.7 g of water was added to the organic dye Varifast Black 3830 sample. The mixture was mixed at 2000 rpm for 1 min × 5 times using a planetary centrifugal mixer (Thinky Corporation, ARE-310), and the dispersion state was visually confirmed. As a result, all of the example compounds were well dispersed, resulting in uniform dispersions. On the other hand, sedimentation was observed immediately after mixing with the comparative example compounds.

[0332] From these results, it was confirmed that the compound was well dispersed even when it was dried after dispersion and then re-dispersed.Furthermore, in addition to the dispersion method of the system in which metal oxides and organic dyes were added to the aqueous solution of the organic ammonium salt of the present invention and dispersed, the system in which water was added to the powder state and dispersed also showed good dispersion, suggesting that the re-dispersion of organic compounds and material treatment agents having hydrogen bond-accepting functional groups, cosmetics, etc. is possible, and that the compound is useful for these applications.

[0333] Furthermore, the amine compounds and acids used as raw materials are compounds registered in the quasi-drug raw material standards, suggesting that they are highly safe and effective for these applications.

[0334] [Table 22]

[0335] 10. Metal oxide dispersibility test 2 For the compounds of each Example and Comparative Example shown in Table 23, 0.25 g of each compound, 0.50 g of water, and 0.10 g of zinc oxide (Ishihara Sangyo Kaisha) were mixed in a planetary centrifugal mixer (Thinky Corporation, ARE-310) at 2000 rpm for 1 minute 5 times, and the dispersion state was then visually confirmed. Those in which the zinc oxide was dispersed were evaluated as ○, and those in which the zinc oxide was not dispersed and had settled were evaluated as ×. The results are shown in Table 23.

[0336] In all of the compounds of the Examples, zinc oxide was dispersed well, and a dispersion was obtained, whereas in the compound of Comparative Example 12, zinc oxide immediately precipitated and did not disperse.

[0337] In other words, the organic ammonium salt of the present invention takes advantage of the structural feature of being composed of a quaternary ammonium cation having many hydrogen bond-donating and coordinating hydroxyl groups, thereby increasing the affinity with the hydrogen bond-accepting oxygen atoms of zinc oxide, and furthermore, it is thought that the affinity between the hydroxyl groups of the quaternary ammonium salt and the coordinating metal atoms of zinc oxide allows for good dispersion of zinc oxide.

[0338] [Table 23]

[0339] 11. Metal oxide dispersibility test 3 For each compound of each Example and Comparative Example shown in Table 24, 0.25 g of each compound, 0.50 g of water, and 0.10 g of barium titanate (Kanto Chemical Co., Ltd.) were mixed in a planetary centrifugal mixer (Thinky Corporation, ARE-310) at 2000 rpm for 1 minute 5 times, and the dispersion state was then visually confirmed. Those in which the barium titanate was dispersed were evaluated as ○, and those in which the barium titanate was not dispersed and had settled were evaluated as ×. The results are shown in Table 24.

[0340] In all of the compounds of the Examples, barium titanate was dispersed well, and a dispersion was obtained, whereas in the compound of Comparative Example 12, the barium titanate immediately precipitated and did not disperse.

[0341] In other words, the organic ammonium salt of the present invention takes advantage of the structural feature of being composed of a quaternary ammonium cation having many hydrogen bond donating and coordinating hydroxyl groups, thereby increasing the affinity with the hydrogen bond accepting oxygen atoms of barium titanate, and furthermore, it is believed that the affinity between the hydroxyl groups of the quaternary ammonium salt and the coordinating metal atoms of zinc oxide results in good dispersion of barium titanate.

[0342] [Table 24]

Claims

1. The following formula (I) 【Chemistry 1】 (wherein R each independently represents a hydroxyalkyl group having one or more hydroxyl groups, the alkyl moiety being linear or branched and having 1 to 10 carbon atoms; and n represents an integer of 1 to 3), and a biological sample. The solid composition comprises an organic ammonium salt that is solid at 25°C and contains an ammonium cation and an anion represented by the formula: (wherein R each independently represents a hydroxyalkyl group having one or more hydroxyl groups, the alkyl moiety being linear or branched and having 1 to 10 carbon atoms; and n represents an integer of 1 to 3), and a biological sample, wherein the mass ratio of the organic ammonium salt to the biological sample is 10:1 to 0.1:

1.

2. 2. The solid composition according to claim 1, wherein R is a monohydroxyalkyl group.

3. 2. The solid composition according to claim 1, wherein at least one of R is a polyhydroxyalkyl group having two or more hydroxyl groups.

4. 2. The solid composition according to claim 1, wherein the anion of the organic ammonium salt is a carboxylate anion, a halide anion, a sulfur-based anion, a fluorine-based anion, a nitrogen oxide-based anion, a phosphorus-based anion, or a cyanide-based anion.

5. The solid composition according to claim 1, wherein the biological sample is a biocatalyst, a protein, or a nucleic acid.

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