Stationary phase for chromatography, and method for producing stationary phase for chromatography

Abstract

Provided are a novel stationary phase for chromatography, and a method for producing a stationary phase for chromatography.　This stationary phase for chromatography contains polymer-carrying particles that contain a hydrophobic polymer and inorganic particles to which the hydrophobic polymer is bonded, the hydrophobic polymer having a repeating unit represented by formula (1). The polymer-carrying particles preferably have a structure represented by formula (2). The average grain diameter of the polymer-carrying particles is preferably 0.1-50 μm. The inorganic particles are preferably silica gel. The stationary phase is preferably for use in reverse phase chromatography.

Description

Stationary phase for chromatography, and method for producing a stationary phase for chromatography.

[0001] This disclosure relates to a stationary phase for chromatography and a method for producing a stationary phase for chromatography. This application claims priority to Japanese Patent Application No. 2025-015943, filed in Japan on 3 February 2025, which is incorporated herein by reference.

[0002] Chromatography is the most effective method for analyzing or separating the components and content of a mixture. It separates different substances by utilizing the intrinsic distribution ratio (also understood as adsorption equilibrium) of substances between a porous solid (stationary phase) spatially fixed in a tube called a column or capillary and a fluid moving through its gaps (mobile phase). Typical examples include gas chromatography and liquid chromatography.

[0003] Liquid chromatography uses a liquid as the mobile phase, and by selecting the appropriate mobile phase, it can be applied to most substances. In liquid chromatography, the most common modes are normal-phase chromatography, which uses a combination of a highly polar stationary phase and a less polar mobile phase, and reversed-phase chromatography, which uses the opposite polarity.

[0004] In recent years, high-performance liquid chromatography (HPLC) has been widely used for the separation and qualitative / quantitative analysis of various substances, and various columns have been developed and are commercially available. For reverse-phase chromatography, for example, C8 columns and C18 columns, which have carbon chains bonded to silica gel, are commonly used. Since the separation characteristics of a substance depend greatly on the column to which it is connected, selecting a column suitable for the separation of the target substance is extremely important in the separation and purification of mixtures.

[0005] Zuvela,P.et al., Chem. Rev., 2019, 119, p.3674-3729

[0006] To separate and purify substances that are difficult to separate with general-purpose C8 and C18 columns, there is a need for a second column that is complementary to C8 and C18 columns, i.e., has different separation characteristics. For example, Non-Patent Literature 1 comprehensively reviews the separation characteristics of columns in which various hydrophobic groups are introduced on the surface of the support (Non-Patent Literature 1). However, the current situation is that this demand has not yet been met.

[0007] This disclosure has been made in view of the above circumstances and is intended to provide a novel stationary phase for chromatography. Another object of this disclosure is to provide a method for producing the above stationary phase.

[0008] In other words, the present disclosure provides a stationary phase for chromatography comprising polymer-supported particles, which include a hydrophobic polymer and inorganic particles to which the hydrophobic polymer is bound, wherein the hydrophobic polymer has repeating units represented by the following formula (1). (In formula (1), R １ X represents hydrogen or an alkyl group having 1 to 6 carbon atoms. ２ is -O-, -NH-, or -N(CH ３ ) indicates R ２ represents an alkylene group having 1 to 20 carbon atoms, a haloalkylene group having 1 to 5 carbon atoms, or a cycloalkylene group having 3 to 10 carbon atoms. Z represents hydrogen, halogen, hydroxyl group, -N(R) ３ ) (Caution ４ ), or a hydroxyalkyl group having 1 to 6 carbon atoms. ３ , R ４ (These represent hydrogen atoms or alkyl groups having 1 to 6 carbon atoms, either identical or different. q represents an integer between 2 and 3000.)

[0009] The polymer-supported particles described above preferably have a structure represented by the following formula (2). (In formula (2), W' represents a single bond or an alkylene group having 1 to 10 carbon atoms. W" represents hydrogen or an alkyl group having 1 to 12 carbon atoms. X represents an amide group, an ester group, an N-alkylamide group having 1 to 3 carbon atoms, an ether group, a sulfoxide group, a sulfone group, a sulfide group, or a phosphate ester group. Y represents an alkylene group having 1 to 30 carbon atoms. V is the same or different and represents an ether group bonded to inorganic particles, an alkoxy group having 1 to 5 carbon atoms, a halogen, an alkyl mercaptyl group having 1 to 20 carbon atoms, a nitrogen-containing group, an allyl group, a 2-methyl-2-propenyl group, or an alkyl group having 1 to 3 carbon atoms. R １ represents hydrogen or an alkyl group having 1 to 6 carbon atoms. X ２ represents -O-, -NH-, or -N(CH ３ ). R ２ represents an alkylene group having 1 to 20 carbon atoms, a haloalkylene group having 1 to 5 carbon atoms, or a cycloalkylene group having 3 to 10 carbon atoms. Z represents hydrogen, a halogen, a hydroxy group, -N(R ３ )(R ４ ), or a hydroxyalkyl group having 1 to 6 carbon atoms. R ３ , R ４ are the same or different and represent hydrogen or an alkyl group having 1 to 6 carbon atoms. p represents an integer from 1 to 10, and q' represents an integer from 10 to 3000. The wavy bond in the formula binds to the surface of the inorganic particles.)

[0010] The average particle diameter of the polymer-supported particles is preferably 0.1 μm to 50 μm.

[0011] In the above stationary phase, the inorganic particles are preferably silica gel.

[0012] The above stationary phase is preferably for reverse phase chromatography.

[0013] In the above formula (1), Z is preferably hydrogen or a halogen.

[0014] The present disclosure also provides a method for obtaining a target substance, including a step of separating and obtaining the target substance using the above stationary phase and a mobile phase.

[0015] The disclosure further provides a method for producing a stationary phase for chromatography, comprising any of the following steps (i) to (v), wherein the hydrophobic (meth)acrylic monomer and / or hydrophobic (meth)acrylamide monomer in any of the following steps (i) to (v) has a structure represented by the following formula (I). (i) A step of radical copolymerizing a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer with surface-modified inorganic particles to which polymerizable functional groups are bonded. (ii) A step of obtaining a polymer by radical polymerization of a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer in the presence of a chain transfer agent having a crosslinkable silyl group at its terminus, and a step of silane coupling of the obtained polymer on the surface of the inorganic particles. (iii) A step of obtaining a polymer by radical copolymerizing a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer with a polymerizable double bond with a silane coupling agent having a polymerizable double bond, and a step of silane coupling of the obtained polymer on the surface of the inorganic particles. (iv) A step of radical copolymerizing a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer with a polymerizable double bond with a silane coupling agent having a polymerizable double bond in the presence of inorganic particles. (v) A step of introducing chain-mobile functional groups onto the surface of inorganic particles and radically polymerizing a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer. (In formula (I), R １ X represents hydrogen or an alkyl group having 1 to 6 carbon atoms. ２ is -O-, -NH-, or -N(CH ３ ) indicates R ２ represents an alkylene group having 1 to 20 carbon atoms, a haloalkylene group having 1 to 5 carbon atoms, or a cycloalkylene group having 3 to 10 carbon atoms. Z represents hydrogen, halogen, hydroxyl group, -N(R) ３ ) (Caution ４ ), or a hydroxyalkyl group having 1 to 6 carbon atoms. ３ , R ４(These represent hydrogen or an alkyl group having 1 to 6 carbon atoms, either identical or different.)

[0016] In the above method for producing the stationary phase, the inorganic particles are preferably silica gel.

[0017] This disclosure provides a novel stationary phase for chromatography.

[0018] This is a chromatogram showing the separation of each sample in HPLC using the stationary phase prepared in Example 1. This is a chromatogram showing the separation of each sample in HPLC using the stationary phase prepared in Example 2. This is a chromatogram showing the separation of each sample in HPLC using the stationary phase prepared in Example 3. This is a chromatogram showing the separation of each sample in HPLC using the stationary phase prepared in Example 4. This is a chromatogram showing the separation of each sample in HPLC using the stationary phase prepared in Example 5. This is a chromatogram showing the separation of each sample in HPLC using the stationary phase prepared in Example 6. This is a chromatogram showing the separation of each sample in HPLC using the stationary phase prepared in Comparative Example 1. This is a chromatogram showing the separation of each sample in HPLC using the stationary phase prepared in Comparative Example 2. This is a chromatogram showing the separation of each sample in HPLC using the stationary phase prepared in Comparative Example 3. This is a chromatogram showing the separation of each sample in HPLC using the stationary phase prepared in Comparative Example 4. These are chromatograms showing the separation of each sample in HPLC using the stationary phase prepared in Comparative Example 5. These are chromatograms showing the separation of each sample in HPLC using the stationary phase prepared in Comparative Example 6. These are chromatograms showing the separation of each sample in HPLC using the stationary phase prepared in Comparative Example 7. These are chromatograms showing the separation of each sample in HPLC using the stationary phase prepared in Comparative Example 8. These are chromatograms showing the separation of each sample in HPLC using the stationary phase prepared in Example 9.

[0019] (Stationary Phase) The stationary phase of this disclosure includes polymer-supported particles. In this specification, "stationary phase" means a material that is fixed inside an analytical instrument (column or capillary) in a chromatography method and distributes the substance to be separated between it and a fluid (mobile phase) that moves in contact with it, thereby leading to separation. If the stationary phase is particles, it may refer to the individual particles or to the aggregate formed by the packing of such particles.

[0020] The polymer-supported particles described above include a hydrophobic polymer and inorganic particles to which the hydrophobic polymer is bound.

[0021] The hydrophobic polymer described above has repeating units represented by the following formula (1). The hydrophobic polymer may have only one type of repeating unit represented by the following formula (1), or it may have two or more types.

[0022] In formula (1), R １ X represents hydrogen or an alkyl group having 1 to 6 carbon atoms. ２ is -O-, -NH-, or -N(CH ３ ) indicates R ２ represents an alkylene group having 1 to 20 carbon atoms, a haloalkylene group having 1 to 5 carbon atoms, or a cycloalkylene group having 3 to 10 carbon atoms. Z represents hydrogen, halogen, hydroxyl group, -N(R) ３ ) (Caution ４ ), or a hydroxyalkyl group having 1 to 6 carbon atoms. ３ , R ４ represents hydrogen or an alkyl group having 1 to 6 carbon atoms, either identical or different. q represents an integer between 2 and 3000.

[0023] The above R １ R represents hydrogen or an alkyl group having 1 to 6 carbon atoms. １ Examples of C1-C6 alkyl groups in include linear or branched alkyl groups such as methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, pentyl, and hexyl groups. １ Hydrogen, methyl groups, and ethyl groups are preferred as the elements.

[0024] The above X ２ is -O-, -NH-, or -N(CH ３ ) indicates.

[0025] The above R ２ This represents an alkylene group having 1 to 20 carbon atoms, a haloalkylene group having 1 to 5 carbon atoms, or a cycloalkylene group having 3 to 10 carbon atoms.

[0026] The above R ２ Examples of alkylene groups having 1 to 20 carbon atoms include methylene group, dimethylethylene group, 1-methylethylene group, 1,1-dimethylmethylene group, trimethylene group, 2-methyltrimethylene group, tetramethylene group, pentamethylene group, 2-methylpentamethylene group, 2-ethylpentamethylene group, 3-methylpentamethylene group, 3-ethylpentamethylene group, hexamethylene group, and 2-methyl Examples of linear or branched alkylene groups include hexamethylene group, 2-ethylhexamethylene group, 3-methylhexamethylene group, 3-ethylhexamethylene group, octamethylene group, nonamethylene group, decamethylene group, undecamethylene group, dodecamethylene group, tridecamethylene group, tetradecamethylene group, pentadecamethylene group, hexadecamethylene group, and octadecamethylene group. Among these, linear or branched alkylene groups having 3 to 20 carbon atoms are preferred, and linear or branched alkylene groups having 8 to 20 carbon atoms are more preferred.

[0027] The above R ２Examples of C1-C5 haloalkylene groups in this context include fluoromethylene group, perfluoromethylene group, fluorodimethylene group, 1,1-difluorodimethylene group, 1,2-difluorodimethylene group, perfluorodimethylene group, 1,1-difluorotrimethylene group, 1,2-difluorotrimethylene group, 1,3-difluorotrimethylene group, 1,1,2,2-tetrafluorotrimethylene group, and perfluorotrimethylene group. Examples include fluoroalkylene groups such as 1,1-difluorotetramethylene group, 1,1,2,2-tetrafluorotetramethylene group, 1,1-difluoropentamethylene group, and 1,1,2,2-tetrafluoropentamethylene group; chloroalkylene groups such as chloromethylene group and chlorodimethylene group; bromoalkylene groups such as bromomethylene group and bromodimethylene group; and iodidealkylene groups such as iodide methylene group and iodide dimethylene group. Among these, fluoroalkylene groups having 1 to 5 carbon atoms are preferred, and fluoroalkylene groups having 1 to 3 carbon atoms are more preferred.

[0028] The above R ２ Examples of cycloalkylene groups having 3 to 10 carbon atoms include cyclopropylene, cyclobutylene, 1,2-cyclopentylene, 1,3-cyclopentylene, cyclopentylidene, 1,2-cyclohexylene, 1,3-cyclohexylene, 1,4-cyclohexylene, and cyclohexylidene. Among these, cycloalkylene groups having 3 to 6 carbon atoms are preferred, and 1,2-cyclohexylene, 1,3-cyclohexylene, and 1,4-cyclohexylene are more preferred.

[0029] The above Z is hydrogen, halogen (preferably fluorine), hydroxyl group, -N(R) ３ ) (Caution ４ ) or a hydroxyalkyl group having 1 to 6 carbon atoms. ３ , R ４ R represents hydrogen or an alkyl group having 1 to 6 carbon atoms, either identical or different. ３ and R ４Examples of C1-C6 alkyl groups in include linear or branched alkyl groups such as methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, pentyl, and hexyl groups. ３ and R ４ The preferred group is hydrogen, a methyl group, an ethyl group, a propyl group, or an isopropyl group, with a methyl group or an ethyl group being more preferred. The above Z is hydrogen, a halogen, or -N(R ３ ) (Caution ４ ) is preferred, and hydrogen or halogen is more preferred.

[0030] The above q represents an integer between 2 and 3000. In one embodiment, the above q is preferably an integer between 10 and 3000, more preferably an integer between 15 and 2500, and even more preferably an integer between 20 and 2000. In another embodiment, the above q is preferably an integer between 2 and 300.

[0031] In the above formula (1), R ２ When R is an alkylene group having 1 to 20 carbon atoms, １ X is hydrogen or an alkyl group having 1 to 6 carbon atoms. ２ is -O-, -NH-, or -N(CH ３ )- and Z is hydrogen, hydroxyl group, -N(R ３ ) (Caution ４ ), or preferably a hydroxyalkyl group having 1 to 6 carbon atoms. In this case, Z is more preferably hydrogen or -N(R ３ ) (Caution ４ ) and more preferably hydrogen.

[0032] In the above formula (1), R ２ When R is a haloalkylene group having 1 to 5 carbon atoms, １ X is hydrogen or an alkyl group having 1 to 6 carbon atoms. ２ is -O-, -NH-, or -N(CH ３ )- and Z is preferably hydrogen or a halogen.

[0033] In the above formula (1), R ２When R is a cycloalkylene group having 3 to 10 carbon atoms, １ X is hydrogen or an alkyl group having 1 to 6 carbon atoms. ２ is -O-, -NH-, or -N(CH ３ )- and Z is hydrogen, -N(R ３ ) (Caution ４ ), or preferably a hydroxyalkyl group having 1 to 6 carbon atoms. In this case, Z is more preferably hydrogen or -N(R ３ ) (Caution ４ ) and more preferably hydrogen.

[0034] Examples of the inorganic particles mentioned above include silica gel, alumina, zirconia, titania, magnesia, glass, kaolin, titanium oxide, silicates, and hydroxyapatite. Among these, silica gel, alumina, and glass are preferred, with silica gel being more preferred. The inorganic particles may be either porous or non-porous inorganic particles. Examples of the porous inorganic particles include porous silica gel, alumina, zirconia, titania, magnesia, glass, kaolin, titanium oxide, silicates, and hydroxyapatite. Among these, porous silica gel, alumina, or glass are preferred, with porous silica gel being more preferred. Examples of the non-porous inorganic particles include non-porous silica gel and non-porous titanium oxide. Among these, non-porous silica gel is preferred. In this specification, the term "silica gel" may be used to refer to a concept that includes both porous silica gel and non-porous silica gel.

[0035] In the polymer-supported particles described above, the inorganic particles support the hydrophobic polymer by chemical bonding with it. When silica gel is used as the inorganic particles, the inorganic particles and the hydrophobic polymer are chemically bonded via the silanol groups present in the silica gel. When a material other than silica gel is used as the inorganic particles, the inorganic particles and the hydrophobic polymer can be chemically bonded via groups introduced by surface treatment of the inorganic particles. Furthermore, by performing the surface treatment, excessive adsorption of the target substance onto the inorganic particles can be suppressed. Examples of surface treatment agents used in the surface treatment include silane coupling agents such as aminopropylsilane, and titanate-based and aluminate-based coupling agents.

[0036] The average particle size of the inorganic particles is, for example, 0.1 to 50 μm, preferably 0.1 to 30 μm, more preferably 1 to 10 μm, and even more preferably 1 to 5 μm. The average pore size of the inorganic particles is, for example, 10 to 10000 Å, preferably 50 to 1000 Å, more preferably 100 to 1000 Å, and even more preferably 100 to 500 Å.

[0037] The specific surface area of ​​the above inorganic particles is, for example, 5 to 1000 m² when porous inorganic particles are used. ２ The amount is / g, preferably 10 to 500m ２ It is / g. When non-porous inorganic particles are used, the above specific surface area is, for example, 0.01 m². ２ / g or more, 5m ２ It is less than / g, preferably 0.01 to 4m ２ The value is / g. When using core-shell type particles, as described later, as the inorganic particles, their specific surface area corresponds to the specific surface area of ​​the shell.

[0038] The average particle size of the polymer-supported particles can be considered to be the same as the average particle size of the inorganic particles used. Generally, when a polymer is supported on inorganic particles, there is no change in the average particle size before and after support that exceeds the margin of error. Therefore, the average particle size of the polymer-supported particles is, for example, 0.1 to 50 μm, preferably 0.1 to 30 μm, more preferably 1 to 10 μm, and even more preferably 1 to 5 μm. The average particle size of the inorganic particles and polymer-supported particles refers to the diameter if they are spherical, and in the case of amorphous particles, it is expressed as the diameter of the sphere that is equal to the volume of the amorphous particle. The average particle size of the inorganic particles and polymer-supported particles can be measured using a device that measures using a microscope image, for example, a Mastersizer 2000E (manufactured by Malvern Panalogical).

[0039] The polymer-supported particles are preferably spherical particles with an aspect ratio of 2 or less, more preferably 1.5 or less. The closer the polymer-supported particles are to a perfect sphere, the better, and the lower limit of the aspect ratio is, for example, 1. The aspect ratio can be measured, for example, by the following method: The sample is randomly scattered on an observation table and observed from directly above with an electron microscope or optical microscope. In any screen where 10 or more primary particles that are not in contact with or overlapping with any other particles are observed, the major axis and minor axis (the length of the longest part perpendicular to the major axis) are determined for each independent primary particle in the screen, and the ratio of the two is taken as the aspect ratio of the individual particle. The aspect ratio used herein is the arithmetic mean of the aspect ratios for all independent primary particles in the screen. Primary particles are particles in which the interfaces between particles can be clearly observed. Normally, observations are performed by appropriately dispersing primary particles on the sample stage to avoid overlapping; however, accidental overlapping is unavoidable, and there are also bulk particles formed by the aggregation of multiple primary particles, but these are excluded from observation.

[0040] The average thickness of the hydrophobic polymer supported on the polymer-supported particles (amount of inorganic particles supported per gram / specific surface area of ​​the inorganic particles) tends to produce a sharp peak, so it is preferable to have a thickness of 0.5 nm or more and 5 nm or less.

[0041] As the inorganic particles mentioned above, core-shell type particles having a non-porous core and a porous shell on its outer surface may be used. In this specification, the core-shell type particles are classified as porous inorganic particles. Here, non-porous refers to the specific surface area (m²) of the core particle's surface as measured by the BET method. ２ Let A be the particle size ( / g), and the surface area (calculated from the particle radius r, 4πr) obtained from the particle diameter of the core particle. ２ The surface area per unit weight (m²) can be calculated from the above. ２ When B is the amount of energy per gram (A-B), then (A-B) / B × 100 is less than 20. On the other hand, porosity refers to a specific surface area of ​​10 mm² as measured by the BET method. ２ This refers to items that are 1 / g or more in weight.

[0042] The ratio of the core thickness to the shell thickness of the above-mentioned core-shell type particle is usually 1:9 to 9:1, and is preferably 4:1 to 2:1 from the viewpoint of ensuring good separation characteristics of the target substance. Here, the core thickness refers to the diameter of the core, and the shell thickness refers to the value obtained by subtracting the core diameter from the diameter of the core-shell type particle and dividing that value by 2. The above ratio can be adjusted by adjusting the shell thickness of the core-shell particle using the method described later.

[0043] The average particle diameter of the core-shell type particles is, for example, 0.1 to 10 μm or less, and preferably 1 to 5 μm. In this specification, the average particle diameter of core-shell type particles refers to the average particle diameter measured by the centrifugal sedimentation method.

[0044] Examples of materials for the core constituting the above-mentioned core-shell type particles include inorganic substances, specifically glass, metals such as titanium and zirconium or their oxides, and clay minerals such as bentonite and mica.

[0045] As the material for the shell constituting the above-mentioned core-shell type particles, from the viewpoint of easily producing the core-shell type particles, it is preferable that the material is a compound obtained by further hydrolysis of a polyalkoxysiloxane obtained by partial hydrolysis of an alkoxysilane. The above-mentioned alkoxysilane is preferably a tetraalkoxysilane, and among these, it is preferable to use tetramethoxylane, tetraethoxysilane, tetrapropoxysilane, and tetrabutoxysilane, and more preferably tetraethoxysilane.

[0046] A method for producing the above-mentioned core-shell type particles can be found in Japanese Patent Publication No. 49-36396. Specifically, first, partial hydrolysis of alkoxysilane is performed to produce polyalkoxysiloxane. Then, the obtained polyalkoxysiloxane is dissolved in a solvent such as ether, acetone, or dichloromethane to prepare a solution of polyalkoxysiloxane. This solution is applied to the core particles or the core particles are immersed in this solution, and then the solvent is removed to deposit the polyalkoxysiloxane as a shell on the surface of the core particles. Subsequently, the deposited polyalkoxysiloxane is subjected to polycondensation (hydrolysis) in the presence of water. This yields core-shell type particles.

[0047] The thickness of the shell constituting the above-mentioned core-shell type particle can be appropriately adjusted within the range of 0.1 to 100 μm, and one method for doing so is to adjust the viscosity of the alkoxysilane that forms the shell. For example, to increase the thickness of the shell, the viscosity of the alkoxysilane can be lowered.

[0048] One method for adjusting the specific surface area and pore diameter of a shell is to adjust the pH of the aqueous solution used when stacking the shells and carrying out polycondensation. For example, to increase the specific surface area and pore diameter, one can consider increasing the pH.

[0049] The average pore diameter of the shell of the above-mentioned core-shell type particles may be, for example, 9 nm or more, or 30 nm or more. The average pore diameter is usually 300 nm or less. When the average pore diameter is within the above range, it is expected that the ligand substance will penetrate into the interior of the shell of the above-mentioned core-shell type particles, contributing to good separation of the target substance. The average pore diameter can be measured by the gas adsorption method. The gas adsorption method is a method in which a gas is adsorbed onto a porous sample by changing the pressure, the amount of adsorption is measured, and the specific surface area, pore volume, pore distribution, etc. are calculated from the adsorption-desorption isotherm obtained by plotting the relative pressure (= ratio of the pressure at the adsorption equilibrium state to the saturated vapor pressure) and the amount of adsorption. JIS Z8831-2 (mesopores and macropores) or JIS Z8831-3 (micropores), which specify the pore size distribution and pore characteristics of powders (solids), can be applied mutatis mutandis.

[0050] The above-mentioned core-shell particles may be "core-shell silica gel" that is sold commercially. In addition, commercially available core-shell silica gel may be used in which the core is made of glass and the shell is made of silica gel (hydrolyzed polyalkoxysiloxane).

[0051] The weight-average molecular weight of the hydrophobic polymer described above is preferably between 1,000 and 5,000,000, from the viewpoint of the polymer's solubility in the solvent, prevention of aggregation of inorganic particles when the polymer is supported on inorganic particles, suppression of dissolution in the mobile phase solvent, and maintenance of the amount of bonding when chemically bonding to inorganic particles, but the optimal point varies depending on the type of polymer. In this specification, the weight-average molecular weight of the polymer is, for example, when the polymer-supported particles have the structure shown in formula (2) or (4) described later, the repeating unit of the main chain is -(CH ２ -CAB) ｋThis refers to the part marked with -. The above weight-average molecular weight can be measured by gel permeation chromatography (GPC) using polystyrene, poly(methyl methacrylate), and poly(ethylene glycol) as standard substances. In the stationary phase manufacturing method described later, if the manufacturing method includes steps (i), (iv), or (v), the polymerization of monomers having hydrophobic groups and bonding with inorganic particles occur simultaneously, so the weight-average molecular weight is estimated from the supernatant of the polymerization solution. In addition, in the stationary phase manufacturing method described later, if the manufacturing method includes steps (ii) or (iii), the weight-average molecular weight of the polymer having hydrophobic groups in the repeating units of the main chain is measured before bonding the polymer to the inorganic particles.

[0052] The loading rate (%) of the hydrophobic polymer in the stationary phase is preferably 1 to 50% by mass, more preferably 3 to 30% by mass, and even more preferably 5 to 25% by mass. The loading rate indicates the proportion of the hydrophobic polymer carried by the polymer-supported particles in the stationary phase. By having the loading rate within the above range, it is possible to appropriately exhibit the adsorption capacity of the hydrophobic polymer while avoiding excessively strong retention or broad peaks. The loading rate can be measured by elemental analysis and can be calculated based on the measurement results of the carbon content of the inorganic particles before the hydrophobic polymer is bonded and the carbon content of the obtained polymer-supported particles, assuming that all carbon other than the carbon contained in the inorganic particles before the hydrophobic polymer is bonded originates from the hydrophobic polymer.

[0053] The polymer-supported particles preferably have a silicon atom between the inorganic particles and the hydrophobic polymer, and more preferably have an Si-O bond.

[0054] The polymer-supported particles described above may have a structure represented by, for example, the following formula (2) or (4).

[0055]

[0056] In formula (2), W' represents a single bond or an alkylene group having 1 to 10 carbon atoms. W" represents hydrogen or an alkyl group having 1 to 12 carbon atoms. X represents an amide group, an ester group, an N-alkylamide group having 1 to 3 carbon atoms, an ether group, a sulfoxide group, a sulfone group, a sulfide group, or a phosphate ester group. Y represents an alkylene group having 1 to 30 carbon atoms. V is the same or different and represents an ether group bonded to inorganic particles, an alkoxy group having 1 to 5 carbon atoms, a halogen, an alkyl mercaptyl group having 1 to 20 carbon atoms, a nitrogen-containing group, an allyl group, a 2-methyl-2-propenyl group, or an alkyl group having 1 to 3 carbon atoms. R １ represents hydrogen or an alkyl group having 1 to 6 carbon atoms. X ２ represents -O-, -NH-, or -N(CH ３ )-. R ２ represents an alkylene group having 1 to 20 carbon atoms, a haloalkylene group having 1 to 5 carbon atoms, or a cycloalkylene group having 3 to 10 carbon atoms. Z represents hydrogen, a halogen, a hydroxy group, -N(R ３ )(R ４ ), or a hydroxyalkyl group having 1 to 6 carbon atoms. R ３ , R ４ are the same or different and represent hydrogen or an alkyl group having 1 to 6 carbon atoms. p represents an integer of 1 to 10, and q' represents an integer of 10 to 3000. The wavy bond in the formula is bonded to the surface of the inorganic particles.

[0057] In the above formula (2), the bonding method of the polymerization unit represented by the above formula (1) and the polymerization unit with p attached may be any of random, alternating, and block.

[0058] The above W' represents a single bond or an alkylene group having 1 to 10 carbon atoms. Examples of linear or branched alkylene groups in the above W' include methylene group, dimethylethylene group, 1-methylethylene group, 1,1-dimethylmethylene group, trimethylene group, 2-methyltrimethylene group, tetramethylene group, pentamethylene group, hexamethylene group, octamethylene group, nonamethylene group, decamethylene group, etc. Among these, linear or branched alkylene groups having 1 to 5 carbon atoms are preferred as the alkylene group having 1 to 10 carbon atoms in the above W', and linear alkylene groups having 1 to 3 carbon atoms are more preferred. The above W' is preferably a single bond, methylene group, dimethylethylene group, or trimethylene group.

[0059] The above W'' represents hydrogen or an alkyl group having 1 to 12 carbon atoms. Examples of C1 to C12 alkyl groups in the above W'' include linear or branched alkyl groups such as methyl group, ethyl group, propyl group, isopropyl group, n-butyl group, isobutyl group, s-butyl group, t-butyl group, pentyl group, hexyl group, 2-ethylhexyl group, nonyl group, and decanyl group. Hydrogen, methyl group, and ethyl group are preferred as the above W''.

[0060] The above X represents an amide group, an ester group, an N-alkylamide group having 1 to 3 carbon atoms, an ether group, a sulfoxide group, a sulfone group, a sulfide group, or a phosphate ester group. Among these, the above X is preferably an amide group, an ester group, or an N-alkylamide group having 1 to 3 carbon atoms.

[0061] The above Y represents an alkylene group having 1 to 30 carbon atoms. Examples of alkylene groups having 1 to 30 carbon atoms in Y include methylene group, dimethylethylene group, 1,1-methylethylene group, trimethylene group, 2-methyltrimethylene group, tetramethylene group, pentamethylene group, 2-methylpentamethylene group, 2-ethylpentamethylene group, 3-methylpentamethylene group, 3-ethylpentamethylene group, hexamethylene group, 2-methylpentamethylene group. Examples of linear or branched alkylene groups include ethylhexamethylene, 2-ethylhexamethylene, 3-methylhexamethylene, 3-ethylhexamethylene, octamethylene, nonamethylene, decamethylene, undecamethylene, dodecamethylene, tridecamethylene, tetradecamethylene, pentamethylene, hexadecamethylene, and octamethylene. Among these, linear or branched alkylene groups having 1 to 10 carbon atoms are preferred, linear or branched alkylene groups having 1 to 5 carbon atoms are more preferred, and methylene, dimethylene, or trimethylene groups are particularly preferred.

[0062] The above V may be the same or different ether group bonded to inorganic particles, a C1-C5 alkoxy group, a halogen, a C1-C20 alkyl mercaptyl group, a nitrogen-containing group, an allyl group, a 2-methyl-2-propenyl group, or a C1-C3 alkyl group. Among these, the above V is preferably an ether group bonded to inorganic particles or a C1-C5 alkoxy group. Examples of the C1-C5 alkoxy group in the above V include a methoxy group or an ethoxy group. Examples of the nitrogen-containing group in the above V include a dimethylamino group, a diethylamino group, a pyrrolidino group, and an imidazolyl group.

[0063] The above R １ R represents hydrogen or an alkyl group having 1 to 6 carbon atoms. １ Examples of C1-C6 alkyl groups in include linear or branched alkyl groups such as methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, pentyl, and hexyl groups. １Examples thereof preferably include hydrogen, a methyl group, and an ethyl group.

[0064] The above X ２ represents -O-, -NH-, or -N(CH ３ ).

[0065] The above R ２ represents an alkylene group having 1 to 20 carbon atoms, a haloalkylene group having 1 to 5 carbon atoms, or a cycloalkylene group having 3 to 10 carbon atoms.

[0066] The above R ２ Examples of the alkylene group having 1 to 20 carbon atoms in the above R include linear or branched alkylene groups such as a methylene group, a dimethylene group, a 1-methylethylene group, a 1,1-dimethylmethylene group, a trimethylene group, a 2-methyltrimethylene group, a tetramethylene group, a pentamethylene group, a 2-methylpentamethylene group, a 2-ethylpentamethylene group, a 3-methylpentamethylene group, a 3-ethylpentamethylene group, a hexamethylene group, a 2-methylhexamethylene group, a 2-ethylhexamethylene group, a 3-methylhexamethylene group, a 3-ethylhexamethylene group, an octamethylene group, a nonamethylene group, a decamethylene group, an undecamethylene group, a dodecamethylene group, a tridecamethylene group, a tetradecamethylene group, a pentadecamethylene group, a hexadecamethylene group, and an octadecamethylene group. Among them, a linear or branched alkylene group having 3 to 20 carbon atoms is preferable, and a linear or branched alkylene group having 8 to 20 carbon atoms is more preferable.

[0067] The above R ２Examples of C1-C5 haloalkylene groups in this context include fluoromethylene group, perfluoromethylene group, fluorodimethylene group, 1,1-difluorodimethylene group, 1,2-difluorodimethylene group, perfluorodimethylene group, 1,1-difluorotrimethylene group, 1,2-difluorotrimethylene group, 1,3-difluorotrimethylene group, 1,1,2,2-tetrafluorotrimethylene group, and perfluorotrimethylene group. Examples include fluoroalkylene groups such as 1,1-difluorotetramethylene group, 1,1,2,2-tetrafluorotetramethylene group, 1,1-difluoropentamethylene group, and 1,1,2,2-tetrafluoropentamethylene group; chloroalkylene groups such as chloromethylene group and chlorodimethylene group; bromoalkylene groups such as bromomethylene group and bromodimethylene group; and iodidealkylene groups such as iodide methylene group and iodide dimethylene group. Among these, fluoroalkylene groups having 1 to 5 carbon atoms are preferred, and fluoroalkylene groups having 1 to 3 carbon atoms are more preferred.

[0068] The above R ２ Examples of cycloalkylene groups having 3 to 10 carbon atoms include cyclopropylene, cyclobutylene, 1,2-cyclopentylene, 1,3-cyclopentylene, cyclopentylidene, 1,2-cyclohexylene, 1,3-cyclohexylene, 1,4-cyclohexylene, and cyclohexylidene. Among these, cycloalkylene groups having 3 to 6 carbon atoms are preferred, and 1,2-cyclohexylene, 1,3-cyclohexylene, and 1,4-cyclohexylene are more preferred.

[0069] The above Z is hydrogen, halogen (preferably fluorine), hydroxyl group, -N(R) ３ ) (Caution ４ ) or a hydroxyalkyl group having 1 to 6 carbon atoms. ３ , R ４ R represents hydrogen or an alkyl group having 1 to 6 carbon atoms, either identical or different. ３ and R ４Examples of C1-C6 alkyl groups in include linear or branched alkyl groups such as methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, pentyl, and hexyl groups. ３ and R ４ The preferred group is hydrogen, a methyl group, an ethyl group, a propyl group, or an isopropyl group, with a methyl group or an ethyl group being more preferred. The above Z is hydrogen, a halogen, or -N(R ３ ) (Caution ４ ) is preferred, and hydrogen or halogen is more preferred.

[0070] The above value of p represents an integer between 1 and 10.

[0071] The above q' represents an integer between 10 and 3000, preferably an integer between 15 and 2500, and more preferably an integer between 20 and 2000.

[0072] In the above equation (2), R ２ When R is an alkylene group having 1 to 20 carbon atoms, １ , X ２ Preferably, the combination of Z is the same as in formula (1) above. In this case, W' is a single bond or an alkylene group having 1 to 10 carbon atoms, W'' is hydrogen or an alkyl group having 1 to 12 carbon atoms, X is an amide group, an ester group, an N-alkylamide group having 1 to 3 carbon atoms, an ether group, a sulfoxide group, a sulfone group, a sulfide group, or a phosphate ester group, and V is the same or different ether group bonded to inorganic particles, an alkoxy group having 1 to 5 carbon atoms, a halogen, an alkyl mercaptyl group having 1 to 20 carbon atoms, a nitrogen-containing group, an allyl group, a 2-methyl-2-propenyl group, or an alkyl group having 1 to 3 carbon atoms. In this case, W' is more preferably a single bond, X is more preferably an amide group, an ester group, or an N-alkylamide group having 1 to 3 carbon atoms, and even more preferably an amide group or an N-alkylamide group having 1 to 3 carbon atoms, and V is more preferably an ether group bonded to inorganic particles or an alkoxy group having 1 to 5 carbon atoms.

[0073] In the above equation (2), R ２When R is a haloalkylene group having 1 to 5 carbon atoms, １ , X ２ Preferably, the combination of Z is the same as in formula (1) above. In this case, W' is a single bond or an alkylene group having 1 to 10 carbon atoms, W'' is hydrogen or an alkyl group having 1 to 12 carbon atoms, X is an amide group, an ester group, an N-alkylamide group having 1 to 3 carbon atoms, an ether group, a sulfoxide group, a sulfone group, a sulfide group, or a phosphate ester group, and V is preferably the same or different ether group bonded to inorganic particles, an alkoxy group having 1 to 5 carbon atoms, a halogen, an alkyl mercaptyl group having 1 to 20 carbon atoms, a nitrogen-containing group, an allyl group, a 2-methyl-2-propenyl group, or an alkyl group having 1 to 3 carbon atoms. In this case, W' is more preferably a single bond, X is more preferably an amide group, an ester group, or an N-alkylamide group having 1 to 3 carbon atoms, and even more preferably an amide group or an N-alkylamide group having 1 to 3 carbon atoms, and Z is more preferably an ether group bonded to inorganic particles or an alkoxy group having 1 to 5 carbon atoms.

[0074] In the above equation (2), R ２ When R is a cycloalkylene group having 3 to 10 carbon atoms, １ , X ２Preferably, the combination of Z is the same as in formula (1) above. In this case, W' is a single bond or an alkylene group having 1 to 10 carbon atoms, W'' is hydrogen or an alkyl group having 1 to 12 carbon atoms, X is an amide group, an ester group, an N-alkylamide group having 1 to 3 carbon atoms, an ether group, a sulfoxide group, a sulfone group, a sulfide group, or a phosphate ester group, and V is the same or different ether group bonded to inorganic particles, an alkoxy group having 1 to 5 carbon atoms, a halogen, an alkyl mercaptyl group having 1 to 20 carbon atoms, a nitrogen-containing group, an allyl group, a 2-methyl-2-propenyl group, or an alkyl group having 1 to 3 carbon atoms. In this case, W' is more preferably a single bond, X is more preferably an amide group, an ester group, or an N-alkylamide group having 1 to 3 carbon atoms, and even more preferably an amide group or an N-alkylamide group having 1 to 3 carbon atoms, and Z is more preferably an ether group bonded to inorganic particles or an alkoxy group having 1 to 5 carbon atoms.

[0075] In formula (4), V' represents an ether group, a C1-C5 alkoxy group, a halogen, a C1-C20 alkyl mercaptyl group, a nitrogen-containing group, an allyl group, a 2-methyl-2-propenyl group, or a C1-C3 alkyl group, either identical or different, bonded to an inorganic particle. Y' represents a single bond or a C1-C30 alkylene group. T' is derived from T, described later, and is a group produced by a chain transfer reaction. R １ , X ２ , R ２ Z is the same as in equation (2) above. r is an integer between 2 and 300. The bonds indicated by the wavy lines in the equation are bonded to the surface of the inorganic particles.

[0076] The above V' represents, either the same or different, an ether group bonded to inorganic particles, a C1-C5 alkoxy group, a halogen, a C1-C20 alkyl mercaptyl group, a nitrogen-containing group, an allyl group, a 2-methyl-2-propenyl group, or a C1-C3 alkyl group. Among these, the above V' is preferably an ether group bonded to inorganic particles or a C1-C5 alkoxy group. Examples of the C1-C5 alkoxy group in the above V' include a methoxy group or an ethoxy group. Examples of the nitrogen-containing group in the above V' include a dimethylamino group, a diethylamino group, a pyrrolidino group, and an imidazolyl group.

[0077] The above Y' represents a single bond or an alkylene group having 1 to 30 carbon atoms. In particular, a single bond or an alkylene group having 1 to 10 carbon atoms is preferred for Y'.

[0078] The above T' is derived from T, which will be described later, and is a group produced by a chain transfer reaction. When T is a C1-C12 haloalkyl group, T' is a halogen-substituted C1-C12 alkylene residue, and when T is a C1-C12 alkyl group having a thiol at its terminus or a C1-C12 alkyl group having a disulfide group within the group, T' is a thioether.

[0079] R in equation (4) above １ , X ２ , R ２ As for Z, R in equation (2) above. １ , X ２ , R ２ Similar examples can be given to Z.

[0080] The above r represents an integer between 2 and 300.

[0081] In the above formula (4), R ２ When R is an alkylene group having 1 to 20 carbon atoms, １ , X ２Preferably, the combination of Z is the same as in formula (1) above. In this case, V' is the same or different ether group bonded to inorganic particles, a carbon-1 to carbon-5 alkoxy group, a halogen, a carbon-1 to carbon-20 alkyl mercaptyl group, a nitrogen-containing group, an allyl group, a 2-methyl-2-propenyl group, or a carbon-1 to carbon-3 alkyl group, Y' is a single bond or a carbon-1 to carbon-30 alkylene group, and T' is a group derived from T and produced by a chain transfer reaction. In this case, V' is more preferably an ether group bonded to inorganic particles or a carbon-1 to carbon-5 alkoxy group.

[0082] In the above formula (4), R ２ When R is a haloalkylene group having 1 to 5 carbon atoms, １ , X ２ Preferably, the combination of Z is the same as in formula (1) above. In this case, V' is the same or different ether group bonded to inorganic particles, a carbon-1 to carbon-5 alkoxy group, a halogen, a carbon-1 to carbon-20 alkyl mercaptyl group, a nitrogen-containing group, an allyl group, a 2-methyl-2-propenyl group, or a carbon-1 to carbon-3 alkyl group, Y' is a single bond or a carbon-1 to carbon-30 alkylene group, and T' is a group derived from T and produced by a chain transfer reaction. In this case, V' is more preferably an ether group bonded to inorganic particles or a carbon-1 to carbon-5 alkoxy group.

[0083] In the above formula (4), R ２ When R is a cycloalkylene group having 3 to 10 carbon atoms, １ , X ２ Preferably, the combination of Z is the same as in formula (1) above. In this case, V' is the same or different ether group bonded to inorganic particles, a carbon-1 to carbon-5 alkoxy group, a halogen, a carbon-1 to carbon-20 alkyl mercaptyl group, a nitrogen-containing group, an allyl group, a 2-methyl-2-propenyl group, or a carbon-1 to carbon-3 alkyl group, Y' is a single bond or a carbon-1 to carbon-30 alkylene group, and T' is a group derived from T and produced by a chain transfer reaction. In this case, V' is more preferably an ether group bonded to inorganic particles or a carbon-1 to carbon-5 alkoxy group.

[0084] Since the above-mentioned stationary phase contains polymer-supported particles in which the hydrophobic polymer and the inorganic particles are bonded, even if a solvent that can dissolve the hydrophobic polymer or a mixed solvent containing it is used as the developing solvent (mobile phase), the polymer will not dissolve, and its function as a stationary phase will not be impaired.

[0085] The above-mentioned stationary phase can be used as a stationary phase for chromatography, and is particularly preferred as a stationary phase for reversed-phase chromatography. The above-mentioned stationary phase can be used by packing it into a chromatography column, for example, by a wet slurry method. Chromatography using the above-mentioned stationary phase may be for analytical purposes or for separation and acquisition purposes.

[0086] Because the stationary phase of this disclosure may have different separation characteristics than general-purpose columns, it can be used in chromatography to separate and obtain substances that are difficult to separate with general-purpose columns.

[0087] The stationary phase of this disclosure can be used to obtain (separately obtain) a target substance. The method for obtaining the target substance includes a step of separating and obtaining the target substance using the stationary phase of this disclosure and a mobile phase. In the step of separating and obtaining the target substance, separation by liquid chromatography is preferred. For example, the mobile phase after passing through a column packed with the stationary phase of this disclosure is separated and obtained using a fraction collector, depending on the separated target substance.

[0088] The column can be of a known size depending on the volume of the sample. The flow rate is not particularly limited, but for example, when expressed as a linear velocity, it is 0.1 to 8 mm / sec, preferably 0.1 to 4 mm / sec. The column temperature is, for example, about 0 to 50°C, preferably about 20 to 45°C.

[0089] The mobile phase is preferably a polar solvent, and may contain additives soluble in the polar solvent. Examples of polar solvents include water, alcohols such as methanol and ethanol, acetonitrile, and mixed solvents thereof. Examples of additives include acids, bases, salts, polyethylene glycol, and the like.

[0090] The above-mentioned stationary phase can be manufactured by the stationary phase manufacturing method described later in this disclosure.

[0091] (Method for manufacturing the stationary phase) The method for manufacturing the stationary phase of this disclosure includes any of the following steps (i) to (v). (i) A step of radical copolymerizing a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer with surface-modified inorganic particles to which polymerizable functional groups are bonded. (ii) A step of obtaining a polymer by radical polymerization of a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer in the presence of a chain transfer agent having a crosslinkable silyl group at its terminus, and a step of silane coupling of the obtained polymer on the surface of the inorganic particles. (iii) A step of obtaining a polymer by radical copolymerizing a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer with a polymerizable double bond with a silane coupling agent having a polymerizable double bond, and a step of silane coupling of the obtained polymer on the surface of the inorganic particles. (iv) A step of radical copolymerizing a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer with a polymerizable double bond with a silane coupling agent having a polymerizable double bond in the presence of inorganic particles. (v) A step of introducing chain-mobile functional groups onto the surface of inorganic particles and radically polymerizing a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer.

[0092] The (meth)acrylic monomer and / or (meth)acrylamide monomer having the hydrophobic group in steps (i) to (v) (hereinafter sometimes simply referred to as "monomer") have the structure represented by the following formula (I).

[0093] In formula (I), R １ X represents hydrogen or an alkyl group having 1 to 6 carbon atoms. ２ is -O-, -NH-, or -N(CH ３ ) indicates R ２represents an alkylene group having 1 to 20 carbon atoms, a haloalkylene group having 1 to 5 carbon atoms, or a cycloalkylene group having 3 to 10 carbon atoms. Z represents hydrogen, halogen, hydroxyl group, -N(R) ３ ) (Caution ４ ), or a hydroxyalkyl group having 1 to 6 carbon atoms. ３ , R ４ These represent hydrogen or an alkyl group having 1 to 6 carbon atoms, either identical or distinct.

[0094] The above R １ R represents hydrogen or an alkyl group having 1 to 6 carbon atoms. １ Examples of C1-C6 alkyl groups in include linear or branched alkyl groups such as methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, pentyl, and hexyl groups. １ Hydrogen, methyl groups, and ethyl groups are preferred as the elements.

[0095] The above X ２ is -O-, -NH-, or -N(CH ３ ) indicates.

[0096] The above R ２ This represents an alkylene group having 1 to 20 carbon atoms, a haloalkylene group having 1 to 5 carbon atoms, or a cycloalkylene group having 3 to 10 carbon atoms.

[0097] The above R ２Examples of alkylene groups having 1 to 20 carbon atoms include methylene group, dimethylethylene group, 1-methylethylene group, 1,1-dimethylmethylene group, trimethylene group, 2-methyltrimethylene group, tetramethylene group, pentamethylene group, 2-methylpentamethylene group, 2-ethylpentamethylene group, 3-methylpentamethylene group, 3-ethylpentamethylene group, hexamethylene group, and 2-methyl Examples of linear or branched alkylene groups include hexamethylene group, 2-ethylhexamethylene group, 3-methylhexamethylene group, 3-ethylhexamethylene group, octamethylene group, nonamethylene group, decamethylene group, undecamethylene group, dodecamethylene group, tridecamethylene group, tetradecamethylene group, pentadecamethylene group, hexadecamethylene group, and octadecamethylene group. Among these, linear or branched alkylene groups having 3 to 20 carbon atoms are preferred, and linear or branched alkylene groups having 8 to 20 carbon atoms are more preferred.

[0098] The above R ２ Examples of C1-C5 haloalkylene groups in this context include fluoromethylene group, perfluoromethylene group, fluorodimethylene group, 1,1-difluorodimethylene group, 1,2-difluorodimethylene group, perfluorodimethylene group, 1,1-difluorotrimethylene group, 1,2-difluorotrimethylene group, 1,3-difluorotrimethylene group, 1,1,2,2-tetrafluorotrimethylene group, and perfluorotrimethylene group. Examples include fluoroalkylene groups such as 1,1-difluorotetramethylene group, 1,1,2,2-tetrafluorotetramethylene group, 1,1-difluoropentamethylene group, and 1,1,2,2-tetrafluoropentamethylene group; chloroalkylene groups such as chloromethylene group and chlorodimethylene group; bromoalkylene groups such as bromomethylene group and bromodimethylene group; and iodidealkylene groups such as iodide methylene group and iodide dimethylene group. Among these, fluoroalkylene groups having 1 to 5 carbon atoms are preferred, and fluoroalkylene groups having 1 to 3 carbon atoms are more preferred.

[0099] The above R ２Examples of cycloalkylene groups having 3 to 10 carbon atoms include cyclopropylene, cyclobutylene, 1,2-cyclopentylene, 1,3-cyclopentylene, cyclopentylidene, 1,2-cyclohexylene, 1,3-cyclohexylene, 1,4-cyclohexylene, and cyclohexylidene. Among these, cycloalkylene groups having 3 to 6 carbon atoms are preferred, and 1,2-cyclohexylene, 1,3-cyclohexylene, and 1,4-cyclohexylene are more preferred.

[0100] The above Z is hydrogen, halogen (preferably fluorine), hydroxyl group, -N(R) ３ ) (Caution ４ ) or a hydroxyalkyl group having 1 to 6 carbon atoms. ３ , R ４ R represents hydrogen or an alkyl group having 1 to 6 carbon atoms, either identical or different. ３ and R ４ Examples of C1-C6 alkyl groups in include linear or branched alkyl groups such as methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, pentyl, and hexyl groups. ３ and R ４ The preferred group is hydrogen, a methyl group, an ethyl group, a propyl group, or an isopropyl group, with a methyl group or an ethyl group being more preferred. The above Z is hydrogen, a halogen, or -N(R ３ ) (Caution ４ ) is preferred, and hydrogen or halogen is more preferred.

[0101] In the above formula (I), R ２ When R is an alkylene group having 1 to 20 carbon atoms, １ , X ２ The preferred combinations of Z are the same as in equation (1) above.

[0102] In the above formula (I), R ２ When R is a haloalkylene group having 1 to 5 carbon atoms, １ , X ２ The preferred combinations of Z are the same as in equation (1) above.

[0103] In the above formula (I), R ２ When R is a cycloalkylene group having 3 to 10 carbon atoms, １ , X ２ The preferred combinations of Z are the same as in equation (1) above.

[0104] A manufacturing method including step (i) is described below. In step (i), a hydrophobic (meth)acrylic monomer and / or hydrophobic (meth)acrylamide monomer are radically copolymerized with surface-modified inorganic particles to which polymerizable functional groups are attached.

[0105] In the manufacturing method including step (i), examples of polymerizable functional groups bonded to the surface-modified inorganic particles include radical polymerizable functional groups, such as vinyl groups, allyl groups, isopropenyl groups, or C4-C12 alkenyl groups having a double bond at the ω position.

[0106] Surface-modified inorganic particles to which the above polymerizable functional groups are attached can be obtained, for example, by silane coupling of the inorganic particles with a compound represented by the following formula (II). Examples of inorganic particles used in silane coupling with the compound represented by the following formula (II) are the same as those of the inorganic particles in the stationary phase of this disclosure.

[0107] W-X-Y-Si(R ａ ) ３－ｎ (Z ２ ) ｎ (II) In formula (II), W represents a vinyl group, an allyl group, an isopropenyl group, or a C4-C12 alkenyl group having a double bond at the ω position. X represents an amide group, an ester group, a C1-C3 N-alkylamide group, an ether group, a sulfoxide group, a sulfone group, a sulfide group, or a phosphate ester group. Y represents an alkylene group having C1-C30. R ａ Each of these independently represents an alkyl group having 1 to 3 carbon atoms. ２ n is a leaving group that can form a bond between the silicon atom and the inorganic particle in formula (II). n is an integer from 1 to 3.

[0108] The above W represents a vinyl group, an allyl group, an isopropenyl group, or an alkenyl group having 4 to 12 carbon atoms and a double bond at the ω position. Among these, the above W is preferably a vinyl group, an allyl group, or an isopropenyl group.

[0109] The above X represents an amide group, an ester group, an N-alkylamide group having 1 to 3 carbon atoms, an ether group, a sulfoxide group, a sulfone group, a sulfide group, or a phosphate ester group. Among these, the above X is preferably an amide group, an ester group, or an N-alkylamide group having 1 to 3 carbon atoms.

[0110] The above Y represents an alkylene group having 1 to 30 carbon atoms. Examples of alkylene groups having 1 to 30 carbon atoms in Y include methylene group, dimethylethylene group, 1,1-methylethylene group, trimethylene group, 2-methyltrimethylene group, tetramethylene group, pentamethylene group, 2-methylpentamethylene group, 2-ethylpentamethylene group, 3-methylpentamethylene group, 3-ethylpentamethylene group, hexamethylene group, 2-methylpentamethylene group. Examples of linear or branched alkylene groups include ethylhexamethylene, 2-ethylhexamethylene, 3-methylhexamethylene, 3-ethylhexamethylene, octamethylene, nonamethylene, decamethylene, undecamethylene, dodecamethylene, tridecamethylene, tetradecamethylene, pentamethylene, hexadecamethylene, and octamethylene. Among these, linear or branched alkylene groups having 1 to 10 carbon atoms are preferred, linear or branched alkylene groups having 1 to 5 carbon atoms are more preferred, and methylene, dimethylene, or trimethylene groups are particularly preferred.

[0111] The above R ａ Each of these independently represents an alkyl group having 1 to 3 carbon atoms. Among them, the above R ａ A methyl group or an ethyl group is preferred.

[0112] The above Z ２Z is a leaving group that can form a bond between the silicon atom in formula (II) and the inorganic particle. For example, if the inorganic particle is silica gel, it can form a bond between the oxygen atom present on the silica gel surface and the silicon atom in formula (II). Commonly used leaving groups that offer a balance between ease of handling and reactivity include alkoxy groups with 1 to 5 carbon atoms, halogens (chlorine, bromine, or iodine), alkyl mercaptyl groups with 1 to 20 carbon atoms, nitrogen-containing groups (e.g., dimethylamino group, diethylamino group, pyrrolidino group, imidazolyl group), allyl group, or 2-methyl-2-propenyl group. ２ Preferably, the leaving group is an alkoxy group having 1 to 5 carbon atoms, and more preferably a methoxy group or an ethoxy group. The reaction conditions (including catalyst addition) can be adjusted depending on the type of leaving group.

[0113] The compound represented by formula (II) above comprises a compound having the structure represented by W in formula (II) above, and Y-SiR in formula (II) above. ３－ｎ Z ｎ It can be obtained by reacting a compound having the structure represented by the above formula (II) with the compound, and the reaction of these compounds produces "-X-" in formula (II) above.

[0114] Compounds having the structure represented by W in the above formula (II) include acrylic acid in which the hydrogen atoms of the vinyl group may be substituted with an alkyl group having 1 to 12 carbon atoms, and halogenated acrylic acid in which the hydrogen atoms of the vinyl group may be substituted with an alkyl group having 1 to 12 carbon atoms.

[0115] In the above formula (II), Y - SiR ３－ｎ Z ｎ Compounds having the structure represented by the above include silane coupling agents that have a group that is a precursor of X as described above and have an alkoxy group with 1 to 5 carbon atoms as a leaving group.

[0116] A stationary phase according to one embodiment of the present disclosure, when produced by a manufacturing method including step (i) described above, is obtained by radical copolymerization of a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer with surface-modified inorganic particles to which the polymerizable functional group is bonded. The copolymerization method involves radical copolymerization of both the (meth)acryloyl group of the monomer and the polymerizable functional group bonded to the inorganic particle, and known reaction conditions can be used.

[0117] The radical polymerization in step (i) can be carried out, for example, by using a small amount of radical generator as a catalyst. As the radical generator, known ones used in radical polymerization reactions can be used, for example, azo compounds and peroxides. The stationary phase of this disclosure obtained by the manufacturing method including step (i) is presumed to contain polymer-supported particles having a structure represented by the following formula (2).

[0118]

[0119] In formula (2), W' represents a single bond or an alkylene group having 1 to 10 carbon atoms. W'' represents hydrogen or an alkyl group having 1 to 12 carbon atoms. X and Y are the same as in formula (II) above. V is the same or different as an ether group, an alkoxy group having 1 to 5 carbon atoms, a halogen, an alkyl mercaptyl group having 1 to 20 carbon atoms, a nitrogen-containing group, an allyl group, a 2-methyl-2-propenyl group, or an alkyl group having 1 to 3 carbon atoms, bonded to the inorganic particle. Bonds indicated by wavy lines in the formula are bonded to the surface of the inorganic particle. １ , X ２ , R ２ Z is the same as in equation (I) above. p is an integer from 1 to 10, and q' is an integer from 10 to 3000. The bonds indicated by the wavy lines in the equation are bonded to the surface of the inorganic particles.

[0120] The above W' represents a single bond or an alkylene group having 1 to 10 carbon atoms. Examples of linear or branched alkylene groups in the above W' include methylene group, dimethylethylene group, 1-methylethylene group, 1,1-dimethylmethylene group, trimethylene group, 2-methyltrimethylene group, tetramethylene group, pentamethylene group, hexamethylene group, octamethylene group, nonamethylene group, decamethylene group, etc. Among these, linear or branched alkylene groups having 1 to 5 carbon atoms are preferred as the alkylene group having 1 to 10 carbon atoms in the above W', and linear alkylene groups having 1 to 3 carbon atoms are more preferred. The above W' is preferably a single bond, methylene group, dimethylethylene group, or trimethylene group.

[0121] The above W'' represents hydrogen or an alkyl group having 1 to 12 carbon atoms. Examples of C1 to C12 alkyl groups in the above W'' include linear or branched alkyl groups such as methyl group, ethyl group, propyl group, isopropyl group, n-butyl group, isobutyl group, s-butyl group, t-butyl group, pentyl group, hexyl group, 2-ethylhexyl group, nonyl group, and decanyl group. Hydrogen, methyl group, and ethyl group are preferred as the above W''.

[0122] Examples of X and Y in equation (2) above are the same as those of X and Y in equation (II) above.

[0123] The above V is the same or different ether group bonded to inorganic particles, or unreacted Z represented by the above formula (II). ２ or R ａThat is, V above represents, either the same or different, an ether group bonded to inorganic particles, a carbon-1 to carbon-5 alkoxy group, a halogen, a carbon-1 to carbon-20 alkyl mercaptyl group, a nitrogen-containing group, an allyl group, a 2-methyl-2-propenyl group, or a carbon-1 to carbon-3 alkyl group. Among these, V is preferably an ether group bonded to inorganic particles or a carbon-1 to carbon-5 alkoxy group. Examples of carbon-1 to carbon-5 alkoxy groups in V include a methoxy group or an ethoxy group. Examples of nitrogen-containing groups in V include a dimethylamino group, a diethylamino group, a pyrrolidino group, and an imidazolyl group.

[0124] R in equation (2) above １ , X ２ , R ２ As for Z, R in equation (I) above is the same for each. １ , X ２ , R ２ Similar examples can be given to Z.

[0125] The above value of p represents an integer between 1 and 10.

[0126] The above q' represents an integer between 10 and 3000, preferably between 15 and 2500, and more preferably between 20 and 2000.

[0127] Next, a manufacturing method including step (ii) will be described. This manufacturing method includes the steps of obtaining a polymer by radical polymerization of a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer in the presence of a chain transfer agent having a crosslinkable silyl group at its terminal, and silane coupling of the obtained polymer on the surface of inorganic particles.

[0128] In step (ii), first, a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer are radically polymerized in the presence of a chain transfer agent having a crosslinkable silyl group at the terminal to obtain a polymer.

[0129] Examples of chain transfer agents having a crosslinkable silyl group at the terminal in step (ii) include compounds represented by the following formula (III). (R ｂ) ３－ｎ (Z ２ ) ｎ Si-Y'-T (III) In formula (III), R ｂ Each of these independently represents an alkyl group having 1 to 5 carbon atoms. ２ represents a leaving group that can form a bond between the silicon atom and the inorganic particle in formula (III). Y' represents a single bond or an alkylene group having 1 to 30 carbon atoms. T represents a chain-transfer functional group. n represents an integer from 1 to 3.

[0130] The above R ｂ Each of these independently represents an alkyl group having 1 to 5 carbon atoms. The above R ｂ Examples of C1-C5 alkyl groups in include linear or branched alkyl groups such as methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, and pentyl groups. ｂ The preferred group is a methyl group, an ethyl group, or a propyl group.

[0131] The above Z ２ Z in equation (II) above is ２ Similar examples can be given.

[0132] The above Y' represents a single bond or an alkylene group having 1 to 30 carbon atoms. Examples of alkylene groups having 1 to 30 carbon atoms in the above Y' include methylene group, dimethylethylene group, 1-methylethylene group, 1,1-dimethylmethylene group, trimethylene group, 2-methyltrimethylene group, tetramethylene group, pentamethylene group, 2-methylpentamethylene group, 2-ethylpentamethylene group, 3-methylpentamethylene group, 3-ethylpentamethylene group, hexamethylene group, 2-methylpentamethylene group. Examples of linear or branched alkylene groups include ethylhexamethylene, 2-ethylhexamethylene, 3-methylhexamethylene, 3-ethylhexamethylene, octamethylene, nonamethylene, decamethylene, undecamethylene, dodecamethylene, tridecamethylene, tetradecamethylene, pentamethylene, hexadecamethylene, and octamethylene. Among these, Y' is preferably a single bond or an alkylene group having 1 to 10 carbon atoms.

[0133] The above T represents a chain-transfer functional group. A chain-transfer functional group is a functional group that actively causes a chain-transfer reaction in polymerization, involving the movement of growth-active species and a restart reaction. The presence of the above chain-transfer functional group makes it possible to control the molecular weight and terminal structure of the polymer to some extent. Examples of the above chain-transfer functional group include C1-C12 haloalkyl groups, C1-C12 alkyl groups having a thiol at the terminal, or C1-C12 alkyl groups having a disulfide group within the group.

[0134] The above n represents an integer between 1 and 3.

[0135] Radical polymerization in step (ii) can be carried out, for example, by using a small amount of radical generator as a catalyst in the presence of the above-mentioned chain transfer agent. In this case, the molecular weight of the polymer can be controlled to some extent by the molar ratio of the chain transfer agent to the monomer. As the above-mentioned radical generator, known ones used in radical polymerization reactions can be used, and examples similar to those of the radical generator in step (i) can be given. By radical polymerization in step (ii), a polymer having the structure represented by the following formula (3) can be obtained.

[0136] In formula (3), R ｂ Z ２ Y' is the same as in formula (III) above. T' is derived from T in formula (III) and is a group produced by the chain transfer reaction. R １ , X ２ , R ２ Z is the same as in equation (I) above. n is an integer from 1 to 3, and r is an integer from 2 to 300.

[0137] The above T' is derived from T in formula (III) and is a group produced by a chain transfer reaction. When T is a C1-C12 haloalkyl group, T' is a halogen-substituted C1-C12 alkylene residue, and when T is a C1-C12 alkyl group having a thiol at its terminus or a C1-C12 alkyl group having a disulfide group within the group, T' is a thioether.

[0138] The above R ｂ Z２ As for Y', R in equation (III) above ｂ Z ２ Similar examples can be given to Y'.

[0139] The above R １ , X ２ , R ２ As for Z, R in equation (I) above １ , X ２ , R ２ Similar examples can be given to Z.

[0140] The above values ​​of n represent integers from 1 to 3, and r represent integers from 2 to 300.

[0141] In the above formula (3), R ２ When R is an alkylene group having 1 to 20 carbon atoms, １ , X ２ The preferred combinations of Z are the same as in equation (1) above, and the preferred combinations of Y' and T' are the same as in equation (4) above. In this case, R ｂ Each of these is an alkyl group having 1 to 5 carbon atoms, and Z ２ It is preferable that Z is an alkoxy group having 1 to 5 carbon atoms, a halogen (chlorine, bromine, or iodine), an alkyl mercaptyl group having 1 to 20 carbon atoms, a nitrogen-containing group, an allyl group, or a 2-methyl-2-propenyl group. ２ The more preferably, it is an alkoxy group having 1 to 5 carbon atoms.

[0142] In the above formula (3), R ２ When R is a haloalkylene group having 1 to 5 carbon atoms, １ , X ２ The preferred combinations of Z are the same as in equation (1) above, and the preferred combinations of Y' and T' are the same as in equation (4) above. In this case, R ｂ Each of these is an alkyl group having 1 to 5 carbon atoms, and Z ２ It is preferable that Z is an alkoxy group having 1 to 5 carbon atoms, a halogen (chlorine, bromine, or iodine), an alkyl mercaptyl group having 1 to 20 carbon atoms, a nitrogen-containing group, an allyl group, or a 2-methyl-2-propenyl group. ２ The more preferably, it is an alkoxy group having 1 to 5 carbon atoms.

[0143] In the above formula (3), R ２ When R is a cycloalkylene group having 3 to 10 carbon atoms, １ , X ２ The preferred combinations of Z are the same as in equation (1) above, and the preferred combinations of Y' and T' are the same as in equation (4) above. In this case, R ｂ Each of these is an alkyl group having 1 to 5 carbon atoms, and Z ２ It is preferable that Z is an alkoxy group having 1 to 5 carbon atoms, a halogen (chlorine, bromine, or iodine), an alkyl mercaptyl group having 1 to 20 carbon atoms, a nitrogen-containing group, an allyl group, or a 2-methyl-2-propenyl group. ２ The more preferably, it is an alkoxy group having 1 to 5 carbon atoms.

[0144] In step (ii), the polymer is obtained by radical polymerization of the monomer, and then the obtained polymer is subjected to silane coupling on the surface of inorganic particles.

[0145] Examples of inorganic particles in process (ii) are similar to those of inorganic particles in the stationary phase of this disclosure.

[0146] As a method for bonding the above polymer and the above inorganic particles by silane coupling, known silane coupling methods can be used. The stationary phase of this disclosure obtained by the manufacturing method including step (ii) above is presumed to contain polymer-supported particles having a structure represented by the following formula (4).

[0147] In formula (4), V' represents the same or different ether group, a carbon-1 to carbon-5 alkoxy group, a halogen, a carbon-1 to carbon-20 alkyl mercaptyl group, a nitrogen-containing group, an allyl group, a 2-methyl-2-propenyl group, or a carbon-1 to carbon-3 alkyl group bonded to the inorganic particle. Y', T', R １ , X ２ , R ２ Z and r are the same as in equation (3) above. The bonds indicated by the wavy lines in the equation are bonded to the surface of the inorganic particles.

[0148] The above V' is the same or different ether group bonded to an inorganic particle, or the unreacted Z in formula (III) above. ２ or R ｂ That is, V' above represents, either the same or different, an ether group bonded to inorganic particles, a carbon-1 to carbon-5 alkoxy group, a halogen, a carbon-1 to carbon-20 alkyl mercaptyl group, a nitrogen-containing group, an allyl group, a 2-methyl-2-propenyl group, or a carbon-1 to carbon-3 alkyl group. Among these, V' is preferably an ether group bonded to inorganic particles or a carbon-1 to carbon-5 alkoxy group. Examples of carbon-1 to carbon-5 alkoxy groups in V' include a methoxy group or an ethoxy group. Examples of nitrogen-containing groups in V' include a dimethylamino group, a diethylamino group, a pyrrolidino group, and an imidazolyl group.

[0149] The above Y', T', R １ , X ２ , R ２ Examples of Z and r are similar to those in equation (3) above.

[0150] Next, a manufacturing method including step (iii) will be described. This manufacturing method includes the steps of obtaining a polymer by radical copolymerizing a (meth)acrylic monomer having a hydrophobic group and / or a (meth)acrylamide monomer having a hydrophobic group with a silane coupling agent having a polymerizable double bond, and silane coupling the obtained polymer on the surface of inorganic particles.

[0151] In step (iii), a polymer is first obtained by radical copolymerizing a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer with a polymerizable double bond using a silane coupling agent.

[0152] Examples of the silane coupling agent having the polymerizable double bond in step (iii) include the compound represented by formula (II).

[0153] Radical polymerization in step (iii) can be generated, for example, by using a small amount of radical generator as a catalyst. At this time, the molecular weight can be controlled by using a suitable chain transfer agent or a known living radical polymerization method. As the radical generator, any known one used in radical polymerization reactions can be used, and examples similar to those of the radical generator in step (i) can be given.

[0154] In step (iii), a polymer is obtained by radical polymerization of the monomer and the silane coupling agent, and then the obtained polymer is subjected to silane coupling on the surface of inorganic particles.

[0155] Examples of inorganic particles in process (iii) are similar to those of inorganic particles in the stationary phase of this disclosure.

[0156] As a method for bonding the above polymer and the above inorganic particles by silane coupling, known silane coupling methods can be used. The stationary phase of this disclosure obtained by the manufacturing method including step (iii) above is presumed to contain polymer-supported particles having the structure represented by formula (2) above.

[0157] Next, a manufacturing method including step (iv) will be described. This manufacturing method includes a step of radical copolymerizing a (meth)acrylic monomer having a hydrophobic group and / or a (meth)acrylamide monomer having a hydrophobic group with a silane coupling agent having a polymerizable double bond in the presence of inorganic particles.

[0158] Examples of the silane coupling agent having the polymerizable double bond in step (iv) include the compound represented by formula (II).

[0159] Examples of inorganic particles in step (iv) are similar to those of inorganic particles in the stationary phase of this disclosure.

[0160] Radical polymerization in step (iv) can be brought about, for example, by using a small amount of radical generator as a catalyst. Known radical generators used in radical polymerization reactions can be used as the radical generator, and examples similar to those of the radical generator in step (i) can be given. The stationary phase according to one embodiment of the present disclosure obtained by a manufacturing method including step (iv) is presumed to contain polymer-supported particles having the structure represented by formula (2) above.

[0161] Next, a manufacturing method including step (v) will be described. This manufacturing method includes the step of introducing a chain-mobile functional group onto the surface of inorganic particles and radical polymerization of a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer.

[0162] In step (v), a method for introducing chain-mobile functional groups to the surface of inorganic particles is obtained by silane coupling a compound having the structure represented by formula (III) above with inorganic particles using a known method.

[0163] Examples of inorganic particles in step (v) are similar to those of inorganic particles in the stationary phase of this disclosure.

[0164] Examples of chain-transfer functional groups in step (v) are similar to those in formula (III) above.

[0165] In step (v), the radical polymerization can be carried out by using a small amount of radical generator as a catalyst in the presence of inorganic particles on which chain-transfer functional groups have been introduced (chemically bonded) to the surface, thereby obtaining polymer-supported particles. As the radical generator, known ones used in radical polymerization reactions can be used, and examples similar to those of the radical generator in step (i) can be given. The stationary phase according to one embodiment of the present disclosure obtained by the manufacturing method including step (v) is presumed to contain polymer-supported particles having the structure represented by formula (4) above.

[0166] The stationary phase obtained by the method for producing the stationary phase of this disclosure can be used as a stationary phase for chromatography, and is particularly preferred to be used as a stationary phase for reversed-phase chromatography.

[0167] An embodiment of the present disclosure will be described in more detail below based on examples, but each embodiment can be combined with any other features disclosed herein. Each configuration and combination thereof in each embodiment is an example, and additions, omissions, substitutions, and other modifications are possible as appropriate without departing from the spirit of the present disclosure. Furthermore, the inventions relating to the present disclosure are not limited by the embodiments or the following examples.

[0168] <Preparation of silica gel with acrylamide groups> Preparation Example 1 188.9 g of silica gel (average particle size 3 μm, average pore size 300 Å) was added to a flask, degassed, and purged with nitrogen. 800 mL of toluene was added to form a slurry, and then 18.9 g of water was added. 25.5 g of N-methyl-N-[3-(trimethoxysilyl)propyl]2-propenamide and 139 mL of toluene were weighed into a beaker and mixed with the silica gel in the flask. The flask was then heated in an oil bath and heated under reflux for 6 hours. After the reaction, the solution was cooled and solid-liquid separation was performed. The obtained powder was washed seven times with 950 mL of methanol and vacuum-dried at 80°C for 8 hours to obtain silica gel with acrylamide groups (hereinafter referred to as acrylamide-treated silica gel). The carbon content of the obtained silica gel was 3.28% by mass. Furthermore, acrylamide-treated silica gel was prepared using the same preparation method with silica gel from different lots, and it was confirmed that the carbon content of the obtained silica gel was 3.40% by mass.

[0169] Preparation Example 2 5.18 g of 4-pyrrolidinopyridine was added to a flask, and after degassing, the flask was purged with nitrogen. 3033 mL of toluene, 138.6 g of N-methylaminopropyltrimethoxysilane, and 144.0 g of triethylamine were added in this order under a nitrogen atmosphere. Then, 152 mL of a toluene solution of 84.2 g of acrylate chloride was added dropwise over approximately 60 minutes under a nitrogen atmosphere. After the dropwise addition, the mixture was heated at 60°C for 1.5 hours to synthesize the crude product N-methyl-N-[3-(trimethoxysilyl)propyl]2-propenamide. The crude product obtained from the above reaction was filtered, and the by-product triethylamine hydrochloride was removed after washing with approximately 350 mL of toluene. The filtrate was collected on 700.0 g of silica gel (average particle size 5 μm, average pore size 300 Å). The silica gel used was pre-dried under vacuum at 150°C for 2 hours and then cooled to room temperature. A toluene solution containing dispersed silica gel was heated in an oil bath at 120°C for 3 hours. After cooling to room temperature, solid-liquid separation was performed, followed by washing with 3.03 kg of toluene once, 2.77 kg of methanol four times, and 2.76 kg of acetone three times. The resulting stationary phase was vacuum-dried overnight at 80°C to obtain acrylamide-treated silica gel. The carbon content of the obtained silica gel was 3.47% by mass.

[0170] Preparation Example 3: 624 g of silica gel (average particle size 5 μm, average pore size 300 Å) was added to a flask, 3100 mL of toluene was added to form a slurry, and then 144.0 g of water was added. 95.0 g of N-methyl-N-[3-(trimethoxysilyl)propyl]2-propenamide was weighed into a beaker and mixed with the silica gel in the flask. The flask was then heated under reflux in an oil bath for 6 hours. After the reaction, the solution was cooled and solid-liquid separation was performed. The obtained powder was washed seven times with 3100 mL of methanol and vacuum-dried at 80°C for 8 hours to obtain acrylamide-treated silica gel. The carbon content of the obtained silica gel was 3.00% by mass.

[0171] <Preparation of Stationary Phase for Chromatography> Example 1 10.0 g of acrylamide-treated silica gel (average particle size 3 μm, average pore size 300 Å) obtained in Preparation Example 1 was placed in a flask, degassed, and purged with nitrogen. 25 mL of toluene, 0.3 mL of dimethyl sulfoxide, 10.4 g of n-octyl acrylate, and 0.35 g of 2,2'-azobis(2,4-dimethylvaleronitrile) were added under a nitrogen atmosphere. The flask was heated in an oil bath to 65°C for 6 hours to allow the copolymerization reaction to proceed. After the copolymerization reaction was complete, the obtained powder was collected by filtration using a glass filter and washed five times with 50 mL of toluene, twice with acetone, and once with methanol. The obtained stationary phase was vacuum-dried at 60°C for 6 hours. The carbon content of the obtained stationary phase was 12.85% by mass. The acrylamide-treated silica gel used as a raw material had a carbon content of 3.28% by mass, which was estimated to indicate that approximately 14.0% by mass of polyalkyl ester was bonded to it. The carbon content was measured by elemental analysis, and the increase was calculated to be entirely due to the polyalkyl ester. The same procedure was followed in the following examples.

[0172] It can be inferred that the stationary phase obtained in Example 1 has a structure represented by the following formula.

[0173] Example 2 15.0 g of acrylamide-treated silica gel (average particle size 3 μm, average pore size 300 Å) obtained in Preparation Example 1 was placed in a flask, degassed, and purged with nitrogen. 23.6 mL of toluene, 0.3 mL of dimethyl sulfoxide, 27.6 g of stearyl acrylate, and 0.52 g of 2,2'-azobis(2,4-dimethylvaleronitrile) were added under a nitrogen atmosphere. The flask was heated in an oil bath at 65°C for 6 hours to allow the copolymerization reaction to proceed. After copolymerization was complete, the obtained powder was collected by filtration using a glass filter and washed five times with 75 mL of toluene, twice with acetone, and once with methanol. The obtained stationary phase was vacuum-dried at 60°C for 6 hours. The carbon content of the obtained stationary phase was 19.42% by mass. Since the carbon content of the acrylamide-treated silica gel used as a raw material was 3.28% by mass, it was estimated that approximately 21.7% by mass of polyalkyl ester was bonded to it.

[0174] It can be inferred that the stationary phase obtained in Example 2 has a structure represented by the following formula.

[0175] Example 3 10.0 g of acrylamide-treated silica gel (average particle size 3 μm, average pore size 300 Å) obtained in Preparation Example 1 was placed in a flask, degassed, and purged with nitrogen. 25 mL of toluene, 0.3 mL of dimethyl sulfoxide, 10.4 g of 2-ethylhexyl acrylate, and 0.35 g of 2,2'-azobis(2,4-dimethylvaleronitrile) were added under a nitrogen atmosphere. The flask was heated in an oil bath to 65°C for 6 hours to allow the copolymerization reaction to proceed. After copolymerization was complete, the obtained powder was collected by filtration using a glass filter and washed five times with 50 mL of toluene, twice with acetone, and once with methanol. The obtained stationary phase was vacuum-dried at 60°C for 6 hours. The carbon content of the obtained stationary phase was 12.34% by mass. Since the carbon content of the acrylamide-treated silica gel used as a raw material was 3.28% by mass, it was estimated that approximately 13.3% by mass of polyalkyl ester was bonded to it.

[0176] It can be inferred that the stationary phase obtained in Example 3 has a structure represented by the following formula.

[0177] Example 4 15.0 g of acrylamide-treated silica gel (average particle size 3 μm, average pore size 300 Å) obtained in Preparation Example 1 was placed in a flask, degassed, and purged with nitrogen. 42 mL of toluene, 0.3 mL of dimethyl sulfoxide, 13.0 g of cyclohexyl acrylate, and 0.52 g of 2,2'-azobis(2,4-dimethylvaleronitrile) were added under a nitrogen atmosphere. The flask was heated in an oil bath at 65°C for 6 hours to allow the copolymerization reaction to proceed. After copolymerization was complete, the obtained powder was collected by filtration using a glass filter and washed five times with 75 mL of toluene, twice with acetone, and once with methanol. The obtained stationary phase was vacuum-dried at 60°C for 6 hours. The carbon content of the obtained stationary phase was 12.57% by mass. Since the carbon content of the acrylamide-treated silica gel used as a raw material was 3.28% by mass, it was estimated that approximately 13.9% by mass of polyalkyl ester was bonded.

[0178] It can be inferred that the stationary phase obtained in Example 4 has a structure represented by the following formula.

[0179] Example 5 15.0 g of acrylamide-treated silica gel (average particle size 3 μm, average pore size 300 Å) obtained in Preparation Example 1 was placed in a flask, degassed, and purged with nitrogen. 42 mL of toluene, 0.3 mL of dimethyl sulfoxide, 16.9 g of 2,2,3,3-tetrafluoropropyl methacrylate, and 0.52 g of 2,2'-azobis(2,4-dimethylvaleronitrile) were added under a nitrogen atmosphere. The flask was heated in an oil bath at 65°C for 6 hours to allow the copolymerization reaction to proceed. After copolymerization was complete, the obtained powder was collected by filtration using a glass filter and washed five times with 75 mL of toluene, twice with acetone, and once with methanol. The obtained stationary phase was vacuum-dried at 60°C for 6 hours. The carbon content of the obtained stationary phase was 11.47% by mass. Since the carbon content of the acrylamide-treated silica gel used as a raw material was 3.28% by mass, it was estimated that approximately 20.9% by mass of polyalkyl ester was bonded to it.

[0180] It can be inferred that the stationary phase obtained in Example 5 has a structure represented by the following formula.

[0181] Example 6 15.0 g of acrylamide-treated silica gel (average particle size 3 μm, average pore size 300 Å) obtained in Preparation Example 1 was placed in a flask, degassed, and purged with nitrogen. 43 mL of toluene, 0.3 mL of dimethyl sulfoxide, 14.2 g of 2,2,2-trifluoromethacrylate, and 0.52 g of 2,2'-azobis(2,4-dimethylvaleronitrile) were added under a nitrogen atmosphere. The flask was heated in an oil bath at 65°C for 6 hours to allow the copolymerization reaction to proceed. After copolymerization was complete, the obtained powder was collected by filtration using a glass filter and washed five times with 75 mL of toluene, twice with acetone, and once with methanol. The obtained stationary phase was vacuum-dried at 60°C for 6 hours. The carbon content of the obtained stationary phase was 10.18% by mass. Since the carbon content of the acrylamide-treated silica gel used as a raw material was 3.28% by mass, it was estimated that approximately 17.2% by mass of polyalkyl ester was bonded to it.

[0182] It can be inferred that the stationary phase obtained in Example 6 has a structure represented by the following formula.

[0183] Example 7 10.0 g of acrylamide-treated silica gel (average particle size 3 μm, average pore size 300 Å) obtained in Preparation Example 1 was placed in a flask, degassed, and purged with nitrogen. 30 mL of 2-methoxyethanol, 0.3 mL of dimethyl sulfoxide, 6.4 g of N-3-dimethylaminopropylacrylamide, and 0.23 g of 2,2'-azobis(2,4-dimethylvaleronitrile) were added under a nitrogen atmosphere. The flask was heated in an oil bath to 65°C for 6 hours to allow the copolymerization reaction to proceed. After copolymerization was complete, the obtained powder was collected by filtration using a glass filter and washed five times with 50 mL of toluene, three times with acetone, and once with methanol. The obtained stationary phase was vacuum-dried at 60°C for 6 hours. The carbon content of the obtained stationary phase was 9.25% by mass. Since the carbon content of the acrylamide-treated silica gel used as a raw material was 3.28% by mass, it was estimated that approximately 10.3% by mass of polyalkylamide was bonded to it.

[0184] It can be inferred that the stationary phase obtained in Example 7 has a structure represented by the following formula.

[0185] Example 8 20.0 g of acrylamide-treated silica gel (average particle size 5 μm, average pore size 300 Å) obtained in Preparation Example 2 was placed in a flask, degassed, and purged with nitrogen. 71 mL of 2-methoxyethanol, 1.4 mL of dimethylacetamide, 8.3 g of 2-hydroxyethylacrylamide, and 0.45 g of 2,2'-azobis(2,4-dimethylvaleronitrile) were added under a nitrogen atmosphere. The flask was heated in an oil bath at 65°C for 6 hours to allow the copolymerization reaction to proceed. After copolymerization was complete, the obtained powder was collected by filtration using a glass filter and washed six times with 160 mL of methanol, twice with acetone, and once with methanol. The obtained stationary phase was vacuum-dried at 60°C for 6 hours. The carbon content of the obtained stationary phase was 7.93% by mass. Since the carbon content of the acrylamide-treated silica gel used as a raw material was 3.47% by mass, it was estimated that approximately 9.2% by mass of polyalkylamide was bonded.

[0186] It can be inferred that the stationary phase obtained in Example 8 has a structure represented by the following formula.

[0187] Example 9 In a beaker, 13.5 g of dodecyl acrylate, 16.0 g of propylene glycol 1-monomethyl ether 2-acetate, 0.3 g of toluene, 0.35 g of 2,2'-azobis(2,4-dimethylvaleronitrile), and 10.0 g of acrylamide-treated silica gel (average particle size 5 μm, average pore size 300 Å) obtained in Preparation Example 3 were added to form a slurry. This slurry was placed in a 100 mL three-necked flask. Then, the slurry remaining in the beaker was added to the three-necked flask with 4.8 g of propylene glycol 1-monomethyl ether 2-acetate. After purging with nitrogen, the flask was heated in an oil bath at 65°C for 6 hours to allow the copolymerization reaction to proceed. After the copolymerization reaction was complete, the obtained powder was collected by filtration and washed five times with 50 mL of toluene and three times with acetone. The obtained stationary phase was vacuum-dried at 60°C for 6 hours.

[0188] It can be inferred that the stationary phase obtained in Example 9 has a structure represented by the following formula.

[0189] (Evaluation of Separation Characteristics: Tanaka Test) In chromatography, the separation characteristics of a column can be evaluated by the Tanaka test (K. Kimata, et al., J. Chromatogr. Sci., Vol. 27, 1989, pp. 721-728). Reverse-phase high-performance liquid chromatography (reverse-phase HPLC) was performed using a 2.1 mmφ × 150 mm column packed with slurry of the stationary phases obtained in Examples 1 to 6, a 0.46 cmφ × 25 cm column packed with slurry of the stationary phase obtained in Example 9, and a commercially available chromatography column as a comparative example, and the separation characteristics of the stationary phases were evaluated by the Tanaka test. The analytical conditions for the Tanaka test performed in this disclosure are as follows. In the Tanaka test conducted in this disclosure, the retention coefficient k' of each compound was calculated from the retention time t of each compound when the retention of uracil was set to t0, and the hydrogen bond recognition ability (k'(Caffeine) / k') was calculated for each column. （Ｐｈｅｎｏｌ） Four statuses were calculated: hydrophobicity retention (k'(Amylbenzene)), hydrophobicity difference recognition ability (k'(Amylbenzene) / k'(Butylbenzene)), and molecular structure recognition ability (k'(Triphenylene) / k'(o-Terphenyl)). The results are shown in Table 1. <Analysis conditions> Mobile phase: methanol / water = 80 / 20 Flow rate: linear velocity = 1.0 mm / sec Detection: UV 254 nm Temperature: 40°C Samples: uracil, caffeine, phenol, butylbenzene, amylbenzene, o-terphenyl, triphenylene

[0190] The commercially available chromatography columns used in each comparative example are as follows: Comparative Example 1: Product name "InertSustainSwift® C8 HP", 0.46φ × 15 cm, particle size 3 μm, manufactured by GL Sciences Co., Ltd. Comparative Example 2: Product name "Inertsil® WP300 C8", 0.46φ × 15 cm, particle size 5 μm, manufactured by GL Sciences Co., Ltd. Comparative Example 3: Product name "L-column3 C18", 0.21φ × 15 cm, particle size 3 μm, manufactured by the Chemicals Evaluation and Research Institute Comparative Example 4: Product name "InertSustain® AQ-C18 HP", 0.21φ × 15 cm, particle size 3 μm, manufactured by GL Sciences Co., Ltd. Comparative Example 5: Product name "InertSustainSwift® C18 Comparative Example 6: Product name "InertSustain® C18", 0.21φ × 15 cm, particle size 3 μm, manufactured by GL Sciences Co., Ltd. Comparative Example 7: Product name "Inertsil® WP300 C18", 0.46φ × 15 cm, particle size 5 μm, manufactured by GL Sciences Co., Ltd. Comparative Example 8: Product name "ACQUITY® UPLC BEH C18 Column", 0.21φ × 15 cm, particle size 1.7 μm, manufactured by Waters Corporation

[0191]

[0192] As shown in Table 1, the stationary phase obtained in Example 1 has a C8 selector in its structure similar to that of commercially available C8 columns, but because it is polymerized and has alkyl ester groups, the structure of the silica gel surface is significantly different. As a result, it was shown that the hydrogen bond recognition ability, hydrophobicity retention, and molecular structure recognition ability differ from those of commercially available C8 columns (Comparative Examples 1 and 2), and the column's separation characteristics are different. Similarly, the stationary phase obtained in Example 2 was shown to have different hydrogen bond recognition ability and molecular structure recognition ability compared to commercially available C18 columns (Comparative Examples 3 to 8), and thus different column separation characteristics. Furthermore, the stationary phases obtained in Examples 3 to 6 and 9 were also shown to have different separation characteristics compared to commercially available C8 and C18 columns.

[0193] Variations of the invention relating to this disclosure are described below. [Note 1] A stationary phase for chromatography comprising polymer-supported particles, the hydrophobic polymer comprising a hydrophobic polymer and inorganic particles to which the hydrophobic polymer is bonded, wherein the hydrophobic polymer has repeating units represented by the following formula (1). (In formula (1), R １ X represents hydrogen or an alkyl group having 1 to 6 carbon atoms. ２ is -O-, -NH-, or -N(CH ３ ) indicates R ２ represents an alkylene group having 1 to 20 carbon atoms, a haloalkylene group having 1 to 5 carbon atoms, or a cycloalkylene group having 3 to 10 carbon atoms. Z represents hydrogen, halogen, hydroxyl group, -N(R) ３ ) (Caution ４ ), or a hydroxyalkyl group having 1 to 6 carbon atoms. ３ , R ４ (wherein is the same or different, representing hydrogen or an alkyl group having 1 to 6 carbon atoms. q is an integer from 2 to 3000.) [Note 2] The stationary phase for chromatography according to Note 1, wherein the polymer-supported particles have a structure represented by the following formula (2). (In formula (2), W' represents a single bond or an alkylene group having 1 to 10 carbon atoms. W'' represents hydrogen or an alkyl group having 1 to 12 carbon atoms. X represents an amide group, an ester group, an N-alkylamide group having 1 to 3 carbon atoms, an ether group, a sulfoxide group, a sulfone group, a sulfide group, or a phosphate ester group. Y represents an alkylene group having 1 to 30 carbon atoms. V represents the same or different ether group bonded to an inorganic particle, an alkoxy group having 1 to 5 carbon atoms, a halogen, an alkyl mercaptyl group having 1 to 20 carbon atoms, a nitrogen-containing group, an allyl group, a 2-methyl-2-propenyl group, or an alkyl group having 1 to 3 carbon atoms. R １ X represents hydrogen or an alkyl group having 1 to 6 carbon atoms. ２ is -O-, -NH-, or -N(CH ３ ) indicates R ２ represents an alkylene group having 1 to 20 carbon atoms, a haloalkylene group having 1 to 5 carbon atoms, or a cycloalkylene group having 3 to 10 carbon atoms. Z represents hydrogen, halogen, hydroxyl group, -N(R)３ ) (Caution ４ ), or a hydroxyalkyl group having 1 to 6 carbon atoms. ３ , R ４ , , and , are the same or different, representing hydrogen or an alkyl group having 1 to 6 carbon atoms. p is an integer from 1 to 10, and q' is an integer from 10 to 3000. The bonds indicated by the wavy lines in the formula are bonded to the surface of the inorganic particles.) [Note 3] The stationary phase for chromatography according to Note 1 or 2, wherein the average particle size of the polymer-supported particles is 0.1 μm to 50 μm (preferably 0.1 to 30 μm, more preferably 1 to 10 μm, and even more preferably 1 to 5 μm). [Note 4] The stationary phase for chromatography according to any one of Notes 1 to 3, wherein the average particle size of the inorganic particles is 0.1 to 50 μm (preferably 0.1 to 30 μm, more preferably 1 to 10 μm, and even more preferably 1 to 5 μm). [Note 5] A stationary phase for chromatography according to any one of Notes 1 to 4, wherein the average pore size of the inorganic particles is 10 to 10,000 Å (preferably 50 to 1,000 Å, more preferably 100 to 1,000 Å, and even more preferably 100 to 500 Å). [Note 6] The inorganic particles are porous inorganic particles, and the specific surface area of ​​the inorganic particles is 5 to 1,000 m². ２ / g (preferably 10 to 500m) ２ A stationary phase for chromatography as described in any one of the appendices 1 to 5, wherein the inorganic particles are non-porous inorganic particles and the specific surface area of ​​the inorganic particles is 0.01 m². ２ / g or more, 5m ２ Less than / g (preferably 0.01 to 4m) ２A stationary phase for chromatography according to any one of Appendix 1 to 5, wherein the ratio of the polymer-supported particles is 2 or less (preferably 1.5 or less). [Appendix 8] A stationary phase for chromatography according to any one of Appendix 1 to 7, wherein the aspect ratio of the polymer-supported particles is 2 or less (preferably 1.5 or less). [Appendix 9] A stationary phase for chromatography according to any one of Appendix 1 to 8, wherein the inorganic particles are silica gel. [Appendix 10] A stationary phase for chromatography according to any one of Appendix 1 to 9, which is for reversed-phase chromatography. [Appendix 11] A stationary phase for chromatography according to any one of Appendix 1 to 10, wherein in formula (1), Z is hydrogen or halogen. [Appendix 12] In formula (1), R ２ is an alkylene group having 1 to 20 carbon atoms (preferably a linear or branched alkylene group having 3 to 20 carbon atoms, more preferably a linear or branched alkylene group having 8 to 20 carbon atoms), R １ is hydrogen or an alkyl group having 1 to 6 carbon atoms (preferably hydrogen, a methyl group, or an ethyl group), X ２ is -O-, -NH-, or -N(CH ３ )- and Z is hydrogen, hydroxyl group, -N(R ３ ) (Caution ４ ), or a hydroxyalkyl group having 1 to 6 carbon atoms (preferably hydrogen or -N(R) ３ ) (Caution ４ A stationary phase for chromatography according to any one of the appendices 1 to 11, wherein R is more preferably hydrogen. [Appendix 13] In formula (1), R ２ is a haloalkylene group having 1 to 5 carbon atoms (preferably a fluoroalkylene group having 1 to 5 carbon atoms, more preferably a fluoroalkylene group having 1 to 3 carbon atoms), and R １ is hydrogen or an alkyl group having 1 to 6 carbon atoms (preferably hydrogen, a methyl group, or an ethyl group), X ２ is -O-, -NH-, or -N(CH ３ )- and Z is hydrogen or halogen, a stationary phase for chromatography according to any one of the appendices 1 to 11. [Appendix 14] In formula (1) above, R ２ R is a cycloalkylene group having 3 to 10 carbon atoms. １is hydrogen or an alkyl group having 1 to 6 carbon atoms (preferably hydrogen, a methyl group, or an ethyl group), X ２ is -O-, -NH-, or -N(CH ３ )- and Z is hydrogen, -N(R ３ ) (Caution ４ ), or a hydroxyalkyl group having 1 to 6 carbon atoms (preferably hydrogen or -N(R) ３ ) (Caution ４ A stationary phase for chromatography according to any one of Appendix 1 to 11, wherein the stationary phase is (1), more preferably hydrogen. [Appendix 15] A method for obtaining a target substance, comprising the step of separating and obtaining the target substance using a stationary phase according to any one of Appendix 1 to 14 and a mobile phase. [Appendix 16] A method for producing a stationary phase for chromatography, comprising any one of the following steps (i) to (v), wherein the hydrophobic (meth)acrylic monomer and / or hydrophobic (meth)acrylamide monomer in any one of the following steps (i) to (v) has a structure represented by the following formula (I). (i) A step of radical copolymerizing a hydrophobic (meth)acrylic monomer and / or hydrophobic (meth)acrylamide monomer with surface-modified inorganic particles to which polymerizable functional groups are bonded. (ii) A step of obtaining a polymer by radical polymerization of a hydrophobic (meth)acrylic monomer and / or hydrophobic (meth)acrylamide monomer in the presence of a chain transfer agent having a crosslinkable silyl group at its terminus, and a step of silane coupling of the obtained polymer on the surface of the inorganic particles. (iii) A step of obtaining a polymer by radical copolymerizing a hydrophobic (meth)acrylic monomer and / or hydrophobic (meth)acrylamide monomer with a silane coupling agent having a polymerizable double bond, and a step of silane coupling of the obtained polymer on the surface of the inorganic particles. (iv) A step of radical copolymerizing a hydrophobic (meth)acrylic monomer and / or hydrophobic (meth)acrylamide monomer with a silane coupling agent having a polymerizable double bond in the presence of inorganic particles. (v) A step of introducing chain-mobile functional groups onto the surface of inorganic particles and radically polymerizing a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer. (In formula (I), R １ X represents hydrogen or an alkyl group having 1 to 6 carbon atoms. ２ is -O-, -NH-, or -N(CH ３ ) indicates R ２ represents an alkylene group having 1 to 20 carbon atoms, a haloalkylene group having 1 to 5 carbon atoms, or a cycloalkylene group having 3 to 10 carbon atoms. Z represents hydrogen, halogen, hydroxyl group, -N(R) ３ ) (Caution ４ ), or a hydroxyalkyl group having 1 to 6 carbon atoms. ３ , R ４ (These represent hydrogen or an alkyl group having 1 to 6 carbon atoms, either identical or different.) [Note 17] The method for producing a stationary phase for chromatography according to Note 16, wherein the inorganic particles are silica gel. [Note 18] The method for producing a stationary phase for chromatography according to Note 16 or 17, wherein the average particle size of the inorganic particles is 0.1 to 50 μm (preferably 0.1 to 30 μm, more preferably 1 to 10 μm, and even more preferably 1 to 5 μm). [Note 19] The method for producing a stationary phase for chromatography according to any one of Notes 16 to 18, wherein the average pore size of the inorganic particles is 10 to 10000 Å (preferably 50 to 1000 Å, more preferably 100 to 1000 Å, and even more preferably 100 to 500 Å). [Note 20] The method for producing a stationary phase for chromatography according to any one of Notes 16 to 18, wherein the inorganic particles are porous inorganic particles, and the specific surface area of ​​the inorganic particles is 5 to 1000 m ２ / g (preferably 10 to 500m) ２ A method for producing a stationary phase for chromatography according to any one of the appendices 16 to 19, wherein the amount is ( / g). [Appendix 21] The inorganic particles are non-porous inorganic particles, and the specific surface area of ​​the inorganic particles is 0.01 m². ２ / g or more, 5m ２ Less than / g (preferably 0.01 to 4m) ２ A method for producing a stationary phase for chromatography according to any one of the appendices 16 to 19, wherein R is / g. [Appendix 22] In formula (I), R ２ is an alkylene group having 1 to 20 carbon atoms (preferably a linear or branched alkylene group having 3 to 20 carbon atoms, more preferably a linear or branched alkylene group having 8 to 20 carbon atoms), R１ is hydrogen or an alkyl group having 1 to 6 carbon atoms (preferably hydrogen, a methyl group, or an ethyl group), X ２ is -O-, -NH-, or -N(CH ３ )- and Z is hydrogen, hydroxyl group, -N(R ３ ) (Caution ４ ), or a hydroxyalkyl group having 1 to 6 carbon atoms (preferably hydrogen or -N(R) ３ ) (Caution ４ A method for producing a stationary phase for chromatography according to any one of the appendices 16 to 21, wherein R is more preferably hydrogen. [Appendix 23] In formula (I), R ２ is a haloalkylene group having 1 to 5 carbon atoms (preferably a fluoroalkylene group having 1 to 5 carbon atoms, more preferably a fluoroalkylene group having 1 to 3 carbon atoms), and R １ is hydrogen or an alkyl group having 1 to 6 carbon atoms (preferably hydrogen, a methyl group, or an ethyl group), X ２ is -O-, -NH-, or -N(CH ３ A method for producing a stationary phase for chromatography according to any one of the appendices 16 to 22, wherein Z is hydrogen or halogen. [Appendix 24] In formula (I), R ２ R is a cycloalkylene group having 3 to 10 carbon atoms. １ is hydrogen or an alkyl group having 1 to 6 carbon atoms (preferably hydrogen, a methyl group, or an ethyl group), X ２ is -O-, -NH-, or -N(CH ３ )- and Z is hydrogen, -N(R ３ ) (Caution ４ ), or a hydroxyalkyl group having 1 to 6 carbon atoms (preferably hydrogen or -N(R) ３ ) (Caution ４ A method for producing a stationary phase for chromatography according to any one of the appendices 16 to 23, wherein the stationary phase is (more preferably hydrogen).

Claims

1. A stationary phase for chromatography comprising polymer-supported particles, each containing a hydrophobic polymer and inorganic particles to which the hydrophobic polymer is bonded, wherein the hydrophobic polymer has repeating units represented by the following formula (1). (In formula (1), R １ X represents hydrogen or an alkyl group having 1 to 6 carbon atoms. ２ is -O-, -NH-, or -N(CH ３ ) indicates R ２ represents an alkylene group having 1 to 20 carbon atoms, a haloalkylene group having 1 to 5 carbon atoms, or a cycloalkylene group having 3 to 10 carbon atoms. Z represents hydrogen, halogen, hydroxyl group, -N(R) ３ ) (Caution ４ ), or a hydroxyalkyl group having 1 to 6 carbon atoms. ３ , R ４ (These represent hydrogen atoms or alkyl groups having 1 to 6 carbon atoms, either identical or distinct. q represents an integer between 2 and 3000.) 2. The stationary phase for chromatography according to claim 1, wherein the polymer-supported particles have a structure represented by the following formula (2). (In formula (2), W' represents a single bond or an alkylene group having 1 to 10 carbon atoms. W" represents hydrogen or an alkyl group having 1 to 12 carbon atoms. X represents an amide group, an ester group, an N-alkylamide group having 1 to 3 carbon atoms, an ether group, a sulfoxide group, a sulfone group, a sulfide group, or a phosphate ester group. Y represents an alkylene group having 1 to 30 carbon atoms. V is the same or different and represents an ether group bonded to inorganic particles, an alkoxy group having 1 to 5 carbon atoms, a halogen, an alkyl mercaptyl group having 1 to 20 carbon atoms, a nitrogen-containing group, an allyl group, a 2-methyl-2-propenyl group, or an alkyl group having 1 to 3 carbon atoms. R １ represents hydrogen or an alkyl group having 1 to 6 carbon atoms. X ２ represents -O-, -NH-, or -N(CH ３ ). R ２ represents an alkylene group having 1 to 20 carbon atoms, a haloalkylene group having 1 to 5 carbon atoms, or a cycloalkylene group having 3 to 10 carbon atoms. Z represents hydrogen, a halogen, a hydroxy group, -N(R ３ )(R ４ ), or a hydroxyalkyl group having 1 to 6 carbon atoms. R ３ , R ４ are the same or different and represent hydrogen or an alkyl group having 1 to 6 carbon atoms. p represents an integer of 1 to 10, and q' represents an integer of 10 to 3000. The bond marked with a wavy line in the formula is bonded to the surface of the inorganic particles.) 3. The stationary phase for chromatography according to claim 1, wherein the average particle size of the polymer-supported particles is 0.1 μm to 50 μm.

4. The stationary phase for chromatography according to claim 1, wherein the inorganic particles are silica gel.

5. The stationary phase for chromatography according to claim 1, which is for reversed-phase chromatography.

6. The stationary phase for chromatography according to claim 1, wherein Z in formula (1) is hydrogen or halogen.

7. A method for obtaining a target substance, comprising the step of separating and obtaining the target substance using a stationary phase and a mobile phase as described in any one of claims 1 to 6.

8. A method for producing a stationary phase for chromatography, comprising any of the following steps (i) to (v), wherein the hydrophobic (meth)acrylic monomer and / or hydrophobic (meth)acrylamide monomer in any of the following steps (i) to (v) has a structure represented by the following formula (I). (i) A step of radical copolymerizing a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer with surface-modified inorganic particles to which polymerizable functional groups are bonded. (ii) A step of obtaining a polymer by radical polymerization of a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer in the presence of a chain transfer agent having a crosslinkable silyl group at its terminus, and a step of silane coupling of the obtained polymer on the surface of the inorganic particles. (iii) A step of obtaining a polymer by radical copolymerizing a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer with a polymerizable double bond with a silane coupling agent having a polymerizable double bond, and a step of silane coupling of the obtained polymer on the surface of the inorganic particles. (iv) A step of radical copolymerizing a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer with a polymerizable double bond with a silane coupling agent having a polymerizable double bond in the presence of inorganic particles. (v) A step of introducing chain-mobile functional groups onto the surface of inorganic particles and radically polymerizing a hydrophobic (meth)acrylic monomer and / or a hydrophobic (meth)acrylamide monomer. (In formula (I), R １ X represents hydrogen or an alkyl group having 1 to 6 carbon atoms. ２ is -O-, -NH-, or -N(CH ３ ) indicates R ２ represents an alkylene group having 1 to 20 carbon atoms, a haloalkylene group having 1 to 5 carbon atoms, or a cycloalkylene group having 3 to 10 carbon atoms. Z represents hydrogen, halogen, hydroxyl group, -N(R) ３ ) (Caution ４ ), or a hydroxyalkyl group having 1 to 6 carbon atoms. ３ , R ４ (These represent hydrogen or an alkyl group having 1 to 6 carbon atoms, either identical or different.) 9. The method for producing a stationary phase for chromatography according to claim 8, wherein the inorganic particles are silica gel.

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