Chromatography stationary phase, separation method, and method for producing chromatography stationary phase

Abstract

A chromatography stationary phase comprising a carrier and a polymer that has an alicyclic group and is supported by the carrier, wherein the carrier has a particle shape, and the polymer that has an alicyclic group has a structural unit represented by formula (1) (in formula (1): R1 represents an adamantyl group, a bornyl group, a norbornyl group, or a menthyl group; the adamantyl group, the bornyl group, the norbornyl group, and the menthyl group each optionally have one or more substituents selected from the group consisting of a deuterium atom, halogen atoms, hydrocarbon groups having 1-3 carbon atoms, and halogenated hydrocarbon groups having 1-3 carbon atoms; and R2 represents a hydrogen atom or a methyl group).

Description

Stationary Phase for Chromatography, Separation Method, and Method for Producing Stationary Phase for Chromatography

[0001] The present disclosure relates to a stationary phase for chromatography, a separation method, and a method for producing a stationary phase for chromatography.

[0002] As the stationary phase for chromatography, a stationary phase formed by chemically bonding an alkyl group such as an octadecyl group (C18, ODS) to a silica carrier is widely used.

[0003] For example, Patent Document 1 discloses a reverse-phase chromatography column containing a silica-based filler to which an alkyl group such as an octadecyl group, an octyl group, a butyl group, or a triacontyl group is bonded.

[0004] Japanese Patent Application Laid-Open No. 2018-096732

[0005] However, even when using a silica-based stationary phase for chromatography to which an alkyl group is chemically bonded, separation may be difficult depending on the substance. Therefore, in order to increase the degree of freedom in stationary phase selection, the development of a stationary phase for chromatography that exhibits a different structural selectivity from conventional products is required.

[0006] An object of the present disclosure is to provide a stationary phase for chromatography that exhibits a different structural selectivity from conventional products.

[0007] To solve the above problems, the inventors of the present disclosure have conducted intensive studies. As a result, they have found that by supporting a polymer having an alicyclic group on a carrier, a stationary phase for chromatography that exhibits a different structural selectivity from conventional products can be obtained. That is, the gist of the present disclosure includes the following.

[0008] 〔1〕 A stationary phase for chromatography having a carrier and a polymer having an alicyclic group supported on the carrier, the carrier having a particle shape, and the polymer having an alicyclic group having a structural unit represented by formula (1). (In the formula, R １is an adamantyl group, a bornyl group, a norbornyl group, or a menthyl group; the adamantyl group, the bornyl group, the norbornyl group, and the menthyl group may each have one or more substituents selected from the group consisting of a deuterium atom, a halogen atom, a hydrocarbon group having 1 to 3 carbon atoms, and a halogenated hydrocarbon group having 1 to 3 carbon atoms; R ２ (This is a hydrogen atom or a methyl group.) [2] R １[1] A chromatographic stationary phase, wherein the alicyclic polymer is an adamantyl group. [3] A chromatographic stationary phase, wherein the polymer having the alicyclic group is supported on the carrier via chemical bonds. [4] A chromatographic stationary phase, wherein the carrier is porous particles or non-porous particles, according to any one of [1] to [3]. [5] A chromatographic stationary phase, wherein the constituent material of the carrier is one or more selected from the group consisting of silica, alumina, zirconia, titania, magnesia, glass, kaolin, silicate, and hydroxyapatite, according to any one of [1] to [4]. [6] A chromatographic stationary phase, wherein it is a reverse-phase chromatographic stationary phase, according to any one of [1] to [5]. [7] A separation method, comprising a separation step of separating a sample by chromatography using the chromatographic stationary phase, wherein the alicyclic stationary phase is according to any one of [1] to [6]. [8] A method for producing a stationary phase for chromatography, comprising a supporting step of supporting a polymer having an alicyclic group on a carrier, wherein the supporting step is any of the following steps (a) to (h), the monomer having an alicyclic group in the following steps (a) to (g) includes a monomer represented by formula (2), and the polymer having an alicyclic group in the following step (h) has a constituent unit represented by formula (1).(a) A step of free radical polymerization of a monomer having an alicyclic group in the presence of a support into which a polymerizable functional group has been introduced. (b) A step of producing a crosslinkable polymer having the crosslinkable silyl group and the alicyclic group by chain transfer polymerization of the monomer having an alicyclic group in the presence of a compound having a crosslinkable silyl group and a chain transfer functional group, and reacting the crosslinkable silyl group of the crosslinkable polymer with the functional group on the surface of the support. (c) A step of producing a polymer-modified silane coupling agent by polymerizing a monomer having an alicyclic group and a silane coupling agent having a polymerizable functional group, and bonding the polymer-modified silane coupling agent to the support by a silane coupling reaction. (d) A step of polymerizing a monomer having an alicyclic group and a silane coupling agent having a polymerizable functional group in the presence of a support. (e) A step of chain transfer polymerization of a monomer having an alicyclic group in the presence of a support into which a chain transfer functional group has been introduced. (f) A step of atom transfer radical polymerization of a monomer having an alicyclic group in the presence of a support into which an atom transfer radical polymerization initiator group has been introduced. (g) A step of producing a crosslinkable polymer having a crosslinkable silyl group and an alicyclic group by atom transfer radical polymerization of a monomer having an alicyclic group in the presence of a compound having a crosslinkable silyl group and an atom transfer radical polymerization initiator group, and reacting the crosslinkable silyl group of the crosslinkable polymer with the functional group of the carrier surface. (h) A step of coating the carrier surface with the polymer having an alicyclic group. (In the formula, R １ is an adamantyl group, a bornyl group, a norbornyl group, or a menthyl group; the adamantyl group, the bornyl group, the norbornyl group, and the menthyl group may each have one or more substituents selected from the group consisting of a deuterium atom, a halogen atom, a hydrocarbon group having 1 to 3 carbon atoms, and a halogenated hydrocarbon group having 1 to 3 carbon atoms; R ２(wherein is a hydrogen atom or a methyl group.) [9] The method for producing a stationary phase for chromatography according to [8], wherein the supporting step is step (f) or step (h).

[10] The method for producing a stationary phase for chromatography according to [8] or [9], wherein the carrier is porous particles or non-porous particles.

[11] The method for producing a stationary phase for chromatography according to any one of [8] to

[10] , wherein the constituent material of the carrier is one or more selected from the group consisting of silica, alumina, zirconia, titania, magnesia, glass, kaolin, silicate, and hydroxyapatite.

[0009] This disclosure offers the advantage of providing a stationary phase for chromatography that exhibits structural selectivity different from conventional products. Furthermore, the problems and effects of this disclosure are not limited to those specifically described above, but also include those that will become clear to those skilled in the art from the entire specification.

[0010] This graph shows the relationship between the logarithm of the separation factor and the water / 1-octanol partition coefficient in the chromatographic stationary phase of Example 3. This graph shows the relationship between the logarithm of the separation factor and the water / 1-octanol partition coefficient in the chromatographic stationary phase of Comparative Example 1. This graph shows the relationship between the logarithm of the separation factor and the water / 1-octanol partition coefficient in the chromatographic stationary phase of Comparative Example 2. This graph shows the relationship between the logarithm of the separation factor and the water / 1-octanol partition coefficient in the chromatographic stationary phase of Comparative Example 3. This graph shows the relationship between the logarithm of the separation factor and the water / 1-octanol partition coefficient in the chromatographic stationary phase of Comparative Example 4.

[0011] The following describes specific embodiments of this disclosure, but each configuration and combination thereof in each embodiment is merely an example, and additions, omissions, substitutions, and other modifications are possible as appropriate, without departing from the spirit of this disclosure. This disclosure is not limited by the embodiments. Furthermore, each aspect disclosed herein can be combined with any other features disclosed herein.

[0012] In this disclosure, "X to Y" indicating a range means "X or greater and Y or less". Furthermore, when numerical ranges expressed as "X to Y" or "X or greater and Y or less" are described in steps (for example, in preferred order), the upper and lower limits of each numerical range can be any combination.

[0013] In this disclosure, any phrase such as "one or more selected from the group consisting of X, Y, and Z" means any of X, Y, Z, a combination of X and Y, a combination of X and Z, a combination of Y and Z, or a combination of X, Y, and Z.

[0014] In this disclosure, descriptions such as "X such as x1, x2, and x3" are examples of X, and do not mean that X is limited to x1, x2, and x3.

[0015] 1. Stationary Phase for Chromatography The first embodiment of this disclosure is a stationary phase for chromatography comprising a carrier having a particle shape and a polymer having alicyclic groups supported on the carrier (hereinafter sometimes simply referred to as "stationary phase"). The stationary phase according to this embodiment has bulky alicyclic groups in the polymer supported on the carrier, and due to these bulky alicyclic groups, it tends to exhibit different structural selectivity from conventional stationary phases, and therefore the range of targets for separation also differs from that of conventional stationary phases. For this reason, it is expected that even for samples that were difficult to separate with conventional stationary phases, high-precision separation can be achieved by using the stationary phase according to this embodiment.

[0016] 1-1. The shape of the carrier (hereinafter sometimes simply referred to as "carrier") that supports the polymer having alicyclic groups is that of particles, preferably spherical particles. In this disclosure, "spherical" includes not only the shape of a perfect sphere, but also shapes whose cross-sectional shape is circular, approximately circular, elliptical, or approximately elliptical, such as oblong spheres and flattened spheres. More specifically, if the aspect ratio of the carrier is 2.0 or less, its shape is considered spherical. The aspect ratio of a spherical carrier is preferably 1.5 or less.

[0017] The aspect ratio of the carrier is measured by the following method: The carrier is randomly scattered on an observation stage and observed from directly above with an electron microscope or optical microscope. In any frame where 10 or more independent primary particles (not in contact with or overlapping with any other particles) are observed, the major axis and minor axis (length of the longest part perpendicular to the major axis) are determined for each independent primary particle in the frame, and the ratio of these two is taken as the aspect ratio of the individual particle. The aspect ratio of the carrier is taken as the arithmetic mean of the aspect ratios of all independent primary particles in the frame. Here, a primary particle is a particle in which the interface between particles can be clearly observed. Normally, observation is performed with the primary particles appropriately dispersed on the sample stage to avoid overlap, but accidental overlap is unavoidable, and there are also bulk particles formed by the aggregation of multiple primary particles, but these are excluded from the observation.

[0018] The particle size (carrier) is not particularly limited, but is preferably 0.1 μm or more and 50 μm or less, more preferably 1 μm or more and 40 μm or less, even more preferably 1 μm or more and 30 μm or less, and particularly preferably 1 μm or more and 10 μm or less.

[0019] In this disclosure, "particle size" means the median diameter corresponding to the 50% cumulative value of the cumulative volume distribution curve measured by a laser diffraction / scattering particle size distribution analyzer, i.e., the volume-based cumulative 50% diameter (D ５０ This means that, if the particles are amorphous, the particle size of the carrier is expressed as the diameter of the sphere that is equal to the particle volume. In this case, the particle size of the carrier is measured using a device that measures using a microscope image, such as the Malvern Mastersizer 2000E.

[0020] Generally, when a polymer is supported on a carrier, there is no change in particle size beyond the margin of error before and after support. Therefore, the particle size of the particles formed by supporting the polymer on a carrier can be considered to be approximately the same as the particle size of the carrier. In other words, the particle size range of the stationary phase, including its preferred range, is the same as the particle size range of the carrier described above.

[0021] Examples of constituent materials for the support include organic materials, inorganic materials, and organic-inorganic hybrid materials. Hereinafter, support materials composed of organic materials, inorganic materials, and organic-inorganic hybrid materials will be referred to as organic support materials, inorganic support materials, and organic-inorganic hybrid support materials, respectively.

[0022] Examples of organic materials that make up the organic carrier include polystyrene, poly(meth)acrylamide, and poly(meth)acrylic acid esters. In this disclosure, "(meth)acrylic" means both or either acrylic and methacrylic. The organic carrier may consist of a single constituent material or of two or more constituent materials.

[0023] Examples of inorganic materials that make up the inorganic support include silica, alumina, zirconia, titania, magnesia, glass, kaolin, silicates, and hydroxyapatite, with silica, alumina, or glass being preferred, and silica being more preferred. The inorganic support may consist of a single constituent material or of two or more constituent materials.

[0024] Examples of organic-inorganic hybrid supports include those formed by a sol-gel reaction between an alkoxysilane and an alkyl-substituted or alkylene-substituted alkoxysilane compound.

[0025] The carrier may be porous particles or non-porous particles, but from the viewpoint of increasing the surface area, porous particles are preferred. Furthermore, the carrier is preferably porous inorganic particles or non-porous inorganic particles, more preferably porous inorganic particles, and particularly preferably porous silica particles. In this disclosure, "porous particles" includes not only carriers in which pores are formed throughout the entire particle, but also surface porous carriers (core-shell carriers) having a structure in which a non-porous core is covered by a porous layer.

[0026] When the carrier is porous particles, the average pore size of the carrier is not particularly limited, but is preferably 1 nm to 1,000 nm, more preferably 5 nm to 500 nm, and even more preferably 10 nm to 100 nm.

[0027] The average pore diameter of the carrier is measured in accordance with JIS Z 8831-2:2010 (Pore size distribution and pore characteristics of powders (solids) - Part 2: Measurement method for mesopores and macropores by gas adsorption; established on April 20, 2010).

[0028] When the carrier is porous particles, the specific surface area of the carrier is not particularly limited, but is preferably 5 m ２ / g or more and 1,000 m ２ / g or less, more preferably 10 m ２ / g or more and 500 m ２ / g or less. When the carrier is a non-porous carrier, the specific surface area of the carrier is not particularly limited, but is preferably 0.005 m ２ / g or more and less than 5 m ２ / g, more preferably 0.01 m ２ / g or more and 4 m ２ / g or less.

[0029] The specific surface area of the carrier is measured by the BET multipoint method using nitrogen gas in accordance with JIS Z 8830:2013 (Method for measuring specific surface area of powders (solids) by gas adsorption; established on July 1, 1990; revised on January 21, 2013).

[0030] 1-2. Polymer having an alicyclic group In this embodiment, the polymer having an alicyclic group supported on the carrier has a structural unit (also referred to as a "repeating unit") represented by the formula (1). The alicyclic group in the polymer having an alicyclic group refers to R １ in the formula (1).

[0031]

[0032] In the formula (1), R １ is an adamantyl group, a bornyl group, a norbornyl group, or a menthyl group, and the adamantyl group may be a 1-adamantyl group or a 2-adamantyl group, but is preferably a 1-adamantyl group (see below). R １The group is preferably an adamantyl group (preferably a 1-adamantyl group), a bornyl group, or a menthyl group, more preferably an adamantyl group (preferably a 1-adamantyl group) or a menthyl group, and even more preferably an adamantyl group (preferably a 1-adamantyl group).

[0033]

[0034] The adamantyl group, bornyl group, norbornyl group, and menthyl group may each have one or more substituents selected from the group consisting of a deuterium atom, a halogen atom, a hydrocarbon group having 1 to 3 carbon atoms, and a halogenated hydrocarbon group having 1 to 3 carbon atoms, but it is preferable that they be unsubstituted.

[0035] Examples of hydrocarbon groups having 1 to 3 carbon atoms include the methyl group, ethyl group, n-propyl group, isopropyl group, and cyclopropyl group.

[0036] A halogenated hydrocarbon group having one to three carbon atoms is a group in which the hydrogen atoms bonded to the carbon atoms of a hydrocarbon group having one to three carbon atoms are replaced with halogen atoms. Examples of halogen atoms include fluorine atoms, chlorine atoms, bromine atoms, and iodine atoms, with fluorine atoms being preferred. The number of halogen atoms in a halogenated hydrocarbon group is not particularly limited, but it is preferable that all the hydrogen atoms bonded to the hydrocarbon group are replaced with halogen atoms. Specific examples of halogenated hydrocarbon groups having one to three carbon atoms include trifluoromethyl groups, pentafluoroethyl groups, heptafluoropropyl groups, and heptafluoroisopropyl groups.

[0037] In formula (1), R ２ This is either a hydrogen atom or a methyl group.

[0038] The following are specific examples of the constituent units represented by equation (1).

[0039]

[0040] The polymer having an alicyclic group may contain other structural units other than the structural unit represented by formula (1), to the extent that it does not impair the effects of the present disclosure. Other structural units are not particularly limited, but include, for example, structural units derived from a carrier into which polymerizable functional groups are introduced, used in step (a) described later; structural units derived from a silane coupling agent having polymerizable functional groups, used in steps (c) and (d) described later; and structural units derived from any polymerizable compound such as styrene compounds and olefin compounds.

[0041] The content of the constituent unit represented by formula (1) in the polymer having an alicyclic group is not particularly limited as long as the effects of this disclosure are obtained, but from the viewpoint of more effectively exhibiting the separation properties due to the polymer having an alicyclic group, it is preferably 80 mol% to 100 mol%, more preferably 90 mol% to 100 mol%, and even more preferably 95 mol% to 100 mol%.

[0042] Polymers having alicyclic groups may have a crosslinked structure. In particular, when the stationary phase is produced by a physicoadsorption method (for example, a method including step (h) described later), it is preferable to perform a crosslinking step after physicoadsorption to introduce a crosslinked structure into the polymer having alicyclic groups. As a result, the polymer having alicyclic groups becomes insoluble and firmly immobilized on the support, which suppresses the dissolution of the polymer having alicyclic groups into the mobile phase when separation is performed by chromatography, and can suppress the deterioration of the stationary phase's function over time.

[0043] Methods for crosslinking polymers containing alicyclic groups include radiation crosslinking by irradiation with gamma rays, X-rays, and electron beams; ultraviolet crosslinking by irradiation with ultraviolet light; thermal crosslinking by heating; crosslinking using compounds containing crosslinkable groups; and combinations thereof. As for the crosslinking method, one or more selected from radiation crosslinking and thermal crosslinking are preferred because they have little effect on the composition of the polymer containing alicyclic groups and have fewer problems with residual reagents. The dose in radiation crosslinking and ultraviolet crosslinking, and the heating temperature and heating time in thermal crosslinking, should be selected according to the degree of crosslinking and the degree of degradation of the polymer containing alicyclic groups.

[0044] The number-average molecular weight (Mn) of the polymer having an alicyclic group is not particularly limited, but is preferably 2,000 to 600,000, more preferably 2,500 to 500,000, and even more preferably 3,000 to 400,000.

[0045] The weight-average molecular weight (Mw) of the polymer having an alicyclic group is not particularly limited, but is preferably 2,000 to 600,000, more preferably 2,500 to 500,000, and even more preferably 3,000 to 400,000, in order to ensure solubility in the solvent and to prevent particle aggregation when the polymer having an amine side chain is supported on a carrier.

[0046] The polydispersity (PDI) of the polymer having an alicyclic group is not particularly limited, but is preferably 1.0 to 5.0, more preferably 1.0 to 3.0, and even more preferably 1.0 to 2.0.

[0047] The number-average molecular weight (Mn) and weight-average molecular weight (Mw) of polymers containing alicyclic groups are calculated in polystyrene equivalents by size exclusion chromatography (SEC). Size exclusion chromatography (SEC) is performed under the conditions described in the Examples section. When the stationary phase is produced by supporting a pre-synthesized polymer containing alicyclic groups on a carrier, as in the method including steps (b), (c), (g), or (h) described later, the number-average molecular weight (Mn) and weight-average molecular weight (Mw) of the polymer containing alicyclic groups refer to the number-average molecular weight (Mn) and weight-average molecular weight (Mw) of this pre-synthesized polymer containing alicyclic groups. On the other hand, when the stationary phase is produced by performing the synthesis of polymers containing alicyclic groups and the support of polymers containing alicyclic groups on a carrier in parallel, as in the method including steps (a), (d), (e), or (f) described later, the number-average molecular weight (Mn) and weight-average molecular weight (Mw) of the polymer containing alicyclic groups are estimated from the supernatant of the polymerization solution.

[0048] The polydispersity (PDI) of a polymer containing an alicyclic group is calculated by dividing the weight-average molecular weight (Mw) calculated by the above method by the number-average molecular weight (Mn).

[0049] The average thickness of the polymer having alicyclic groups supported on the carrier (amount of polymer supported per gram of carrier / specific surface area of ​​the carrier) is not particularly limited, but is preferably 5 nm to 25 nm, and more preferably 10 nm to 20 nm. When the average thickness of the polymer having alicyclic groups is within the above range, a sharp peak tends to be obtained, which is preferable.

[0050] The content of the polymer having alicyclic groups in the stationary phase, in which the polymer having alicyclic groups is supported on a carrier, is not particularly limited, but is preferably 10% by mass or more and 45% by mass or less, more preferably 20% by mass or more and 30% by mass or less, and even more preferably 25% by mass or more and 30% by mass or less. By keeping the content of the polymer having alicyclic groups within the above range, it is possible to suppress the broadening of the peak width while appropriately expressing the adsorption capacity of the polymer having alicyclic groups. The content of the polymer having alicyclic groups in the stationary phase is measured by elemental analysis.

[0051] The content of alicyclic polymers in the stationary phase is measured by elemental analysis. Based on the measurement results of the carbon content of the support that does not support alicyclic polymers and the carbon content of the stationary phase, the content of alicyclic polymers in the stationary phase is calculated by assuming that all carbon other than the carbon contained in the support that does not support alicyclic polymers originates from the alicyclic polymers.

[0052] However, if the support does not contain carbon and remains on the boat as combustion ash after elemental analysis (for example, if the support is an inorganic support such as silica particles), the content of polymers having alicyclic groups in the stationary phase is evaluated by the degree of organic modification. The degree of organic modification is calculated based on the results of elemental analysis using the following formula. The degree of organic modification of the stationary phase according to this embodiment is not particularly limited, but is preferably 10% by mass or more and 45% by mass or less, more preferably 20% by mass or more and 30% by mass or less, and even more preferably 25% by mass or more and 30% by mass or less. By keeping the degree of organic modification of the stationary phase within the above range, it is possible to suppress the broadening of the peak width while appropriately expressing the adsorption capacity of polymers having alicyclic groups.

[0053]

[0054] In this disclosure, elemental analysis is performed by combustion. For elemental analysis, an elemental analyzer such as the Yanaco CHN Coder MT-5 can be used.

[0055] The manner in which the polymer having alicyclic groups is supported on the support is not particularly limited, but it is preferable that the polymer having alicyclic groups is supported on the support via chemical bonds, or that the polymer having alicyclic groups is physically adsorbed (coated) on the support. It is more preferable that the polymer having alicyclic groups is supported on the support via chemical bonds. This can further improve the separation performance of the stationary phase. Furthermore, when performing separation by chromatography, even if the mobile phase contains a good solvent for the polymer having alicyclic groups, it is possible to prevent the polymer having alicyclic groups from dissolving in the solvent of the mobile phase and being removed from the stationary phase, thereby suppressing the deterioration of the stationary phase's function over time.

[0056] 1-3. Method for Manufacturing a Stationary Phase for Chromatography The method for manufacturing the stationary phase is selected according to the support configuration described above. If the stationary phase according to this embodiment is a polymer having alicyclic groups supported on a support via chemical bonds, the stationary phase according to this embodiment can be manufactured by a method including any of the following steps (a) to (g). When the stationary phase is manufactured by a method including steps (a), (d), (e), or (f), the synthesis of the polymer having alicyclic groups and the support of the polymer having alicyclic groups occur in parallel. On the other hand, when the stationary phase is manufactured by a method including steps (b), (c), or (g), the polymer having alicyclic groups is synthesized once and then supported on the support via chemical bonds. On the other hand, if the stationary phase according to this embodiment is a polymer having alicyclic groups physically adsorbed on a support, the stationary phase according to this embodiment can be manufactured by a method including the following step (h).

[0057] (a) A step of free radical polymerization of a monomer having an alicyclic group in the presence of a support into which a polymerizable functional group has been introduced. (b) A step of producing a crosslinkable polymer having the crosslinkable silyl group and the alicyclic group by chain transfer polymerization of the monomer having an alicyclic group in the presence of a compound having a crosslinkable silyl group and a chain transfer functional group, and reacting the crosslinkable silyl group of the crosslinkable polymer with the functional group on the surface of the support. (c) A step of producing a polymer-modified silane coupling agent by polymerizing a monomer having an alicyclic group and a silane coupling agent having a polymerizable functional group, and bonding the polymer-modified silane coupling agent to the support by a silane coupling reaction. (d) A step of polymerizing a monomer having an alicyclic group and a silane coupling agent having a polymerizable functional group in the presence of a support. (e) A step of chain transfer polymerization of a monomer having an alicyclic group in the presence of a support into which a chain transfer functional group has been introduced. (f) A step of atom transfer radical polymerization of a monomer having an alicyclic group in the presence of a support into which an atom transfer radical polymerization initiator group has been introduced. (g) A step of producing a crosslinkable polymer having the crosslinkable silyl group and the alicyclic group by atom transfer radical polymerization of a monomer having an alicyclic group in the presence of a compound having a crosslinkable silyl group and an atom transfer radical polymerization initiator group, and reacting the crosslinkable silyl group of the crosslinkable polymer with the functional group of the carrier surface. (h) A step of coating the carrier surface with the polymer having an alicyclic group.

[0058] In either method, the stereoregularity of polymers containing alicyclic groups can be controlled by selecting the polymerization temperature, polymerization solvent, and additives during polymerization.

[0059] The following describes step (a). Examples of polymerizable functional groups used in step (a) of the carrier into which polymerizable functional groups are introduced include vinyl groups, allyl groups, isopropenyl groups, and alkenyl groups having 4 to 12 carbon atoms and a double bond at the ω position, which are groups having an ethylenically unsaturated bond. Preferably, the group is a vinyl group, an allyl group, or an isopropenyl group.

[0060] For example, when porous silica particles or non-porous silica particles (hereinafter, the term "silica particles" may be used to refer to both) are used as a carrier, a carrier in which polymerizable functional groups are chemically bonded to the silica particles via the silanol groups present in the silica particles can be used as a "carrier into which polymerizable functional groups have been introduced."

[0061] When using a support other than silica particles, surface treatment of the support can suppress excessive adsorption of the sample onto the support itself, and can also chemically bond with polymerizable functional groups via groups introduced by the surface treatment. Examples of surface treatment agents include silane coupling agents such as 3-aminopropyltrimethoxysilane; titanium coupling agents; and aluminate coupling agents.

[0062] A support into which polymerizable functional groups are introduced can be obtained, for example, by modifying the surface of the support (preferably silica particles) using a silane coupling reaction with a silane coupling agent represented by formula (I). W-X-Y-SiR ３－ｎ Z ｎ (I)

[0063] In formula (I), W represents a polymerizable functional group (radical polymerizable functional group). The polymerizable functional group is preferably a group having an ethylenically unsaturated bond, more preferably a vinyl group, an allyl group, an isopropenyl group, or an alkenyl having 4 to 12 carbon atoms and a double bond at the ω position, and even more preferably a vinyl group, an allyl group, or an isopropenyl group.

[0064] In formula (I), 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. Preferably, X is an amide group, an N-alkylamide group having 1 to 3 carbon atoms, or an ester group.

[0065] In formula (I), Y represents an alkylene group having 1 to 30 carbon atoms. Y is preferably an alkylene group having 1 to 10 carbon atoms, more preferably an alkylene group having 1 to 5 carbon atoms, and even more preferably a methylene group, an ethylene group, or a trimethylene group.

[0066] In formula (I), each R independently represents an alkyl group having 1 to 5 carbon atoms. R is preferably an alkyl group having 1 to 3 carbon atoms, more preferably a methyl group, an ethyl group, or an n-propyl group, and even more preferably a methyl group or an ethyl group.

[0067] In formula (I), Z independently represents an alkoxy group having 1 to 5 carbon atoms, a halogen atom, an alkylthio group having 1 to 20 carbon atoms, an amino group, or an allyl group having 1 to 5 carbon atoms. Examples of alkoxy groups having 1 to 5 carbon atoms include methoxy and ethoxy groups. Examples of halogen atoms include chlorine, bromine, and iodine atoms. Examples of alkylthio groups having 1 to 20 carbon atoms include methylthio and ethylthio groups. Examples of amino groups include dimethylamino, diethylamino, pyrrolidino, and imidazolyl groups. Examples of allyl groups having 1 to 5 carbon atoms include alkyl-substituted or unsubstituted allyl groups, such as allyl and 2-methyl-2-propenyl groups. Z is preferably an alkoxy group having 1 to 5 carbon atoms, and more preferably a methoxy or ethoxy group, in terms of a good balance between ease of handling and reactivity.

[0068] In formula (I), n represents an integer between 1 and 3, inclusive. Preferably, n is 3.

[0069] The compound represented by formula (I) may be a commercially available compound, but it may also be a compound containing W and -Y-SiR ３－ｎ Z ｎ It can also be produced by reacting it with a compound having a group represented by . Furthermore, the reaction of these compounds with each other produces "-X-" in formula (I).

[0070] Examples of compounds containing W include acrylic acid, in which the hydrogen atoms on the carbon atoms bonded to the vinyl group may be substituted with alkyl groups having 1 to 12 carbon atoms; and its halides; and the like.

[0071] -Y-SiR ３－ｎ Z ｎExamples of compounds having the group represented by include silane coupling agents having an alkoxy group with 1 to 5 carbon atoms as Z.

[0072] The carrier into which polymerizable functional groups are introduced is preferably a surface-modified silica particle obtained by a silane coupling reaction between a compound represented by formula (I) and silica particles.

[0073] In step (a), when a polymer having an alicyclic group is supported on a carrier, the polymer having an alicyclic group is produced by free radical polymerization of the polymerizable functional group of the monomer having an alicyclic group and the polymerizable functional group introduced on the carrier. Any conditions can be used as reaction conditions for free radical polymerization, such as conditions known in the field of polymer synthesis and conditions similar thereto.

[0074] Monomers having an alicyclic group are represented by formula (2). Constituent units represented by formula (1) are produced from monomers represented by formula (2).

[0075]

[0076] In formula (2), R １ and R ２ These are R in equation (1), respectively. １ and R ２ This is synonymous with the same thing, and the preferred form is also the same.

[0077] Specific examples of monomers having an alicyclic group represented by formula (2) include 1-adamantyl(meth)acrylate, 2-adamantyl(meth)acrylate, bornyl(meth)acrylate, isobornyl(meth)acrylate, and menthyl(meth)acrylate, preferably one or more selected from the group consisting of 1-adamantyl(meth)acrylate, bornyl(meth)acrylate, and menthyl(meth)acrylate, and more preferably 1-adamantyl(meth)acrylate.

[0078] Free radical polymerization is preferably carried out in the presence of a catalytic amount of a radical generator. Any known radical generator or similar radical generator can be used as the radical generator. Examples of known radical generators include azo compounds and peroxides.

[0079] Formula (II) shows a structure that is presumed to be a preferred embodiment of the stationary phase obtained by supporting a polymer having an alicyclic group on a carrier in step (a). In formula (II), the polymer may be a block copolymer or a random copolymer.

[0080]

[0081] In formula (II), W' and W'' represent groups formed by the polymerization of W, the polymerizable functional group in formula (I), respectively. Specific examples of W' include single bonds and alkylene groups having 1 to 10 carbon atoms, preferably single bonds or methylene groups. Specific examples of W'' include hydrogen atoms or methyl groups.

[0082] In formula (II), X and Y are the same as X and Y in formula (I), and the preferred embodiments are the same.

[0083] In formula (II), Z' represents a linking group or single bond formed between the silicon atom in formula (I) and the support by a silane coupling reaction. For example, if Z in formula (I) is an alkoxy group having 1 to 5 carbon atoms and the support is silica particles, then Z' is -O-.

[0084] In formula (II), V is a group bonded to the support via Z', Z, R, or a group resulting from the coupling of compounds represented by formula (I). Z and R are the same as Z and R in formula (I), and the preferred embodiment is the same. When V is a group resulting from the coupling of compounds represented by formula (I), n in formula (I) is 2 or 3.

[0085] In the compound represented by formula (II), when n=1, V=R; when n=2, the total proportion of unreacted Z and R relative to the total number of V is between 5% and 100%; and when n=3, the proportion of unreacted Z relative to the total number of V is between 0% and 100%.

[0086] In formula (II), R １ and R ２ These are R in equation (1), respectively. １ and R２ This is synonymous with the same thing, and the preferred form is also the same.

[0087] In formula (II), p represents a value between 1 and 10. Preferably, p is between 1 and 5.

[0088] In formula (II), q represents a value between 10 and 3,000. Preferably, q is between 15 and 2,500, and more preferably between 20 and 2,000.

[0089] The following describes step (b). Examples of compounds having a crosslinkable silyl group and a chain-transfer functional group used in step (b) include the compound represented by formula (III). A crosslinkable silyl group is a group that, depending on the reaction conditions, can undergo homocoupling reactions and reactions with functional groups on the support surface. In formula (III), the crosslinkable silyl group is -SiR ３－ｎ Z ｎ This is a silyl group represented by T-Y-SiR. ３－ｎ Z ｎ (III)

[0090] In formula (III), T is a chain-transfer functional group. A chain-transfer functional group is a functional group that actively causes a chain-transfer reaction in a radical polymerization reaction, involving the movement of growth-active species and a restart reaction. The presence of a chain-transfer functional group allows for some degree of control over the molecular weight and terminal structure of polymers containing alicyclic groups. Preferred chain-transfer functional groups include halogenated alkyl groups having 1 to 12 carbon atoms, alkyl groups having 1 to 12 carbon atoms with a thiol group at the terminal, and alkyl groups having 1 to 12 carbon atoms with a disulfide group in the group. Examples of halogens in halogenated alkyl groups having 1 to 12 carbon atoms include chlorine, bromine, and iodine. Preferred halogenated alkyl groups having 1 to 3 carbon atoms are preferred.

[0091] In formula (III), Y, R, Z, and n are the same as Y, R, Z, and n in formula (I), respectively, and the preferred embodiments are the same.

[0092] The monomer having an alicyclic group used in step (b) is the same monomer having an alicyclic group used in step (a), that is, the monomer represented by formula (2).

[0093] Chain transfer polymerization is preferably carried out in the presence of a catalytic amount of radical generator. Any known radical generator or similar radical generator can be used. Examples of known radical generators include azo compounds and peroxides. In this case, the molecular weight can be controlled to some extent by the molar ratio of the chain transfer agent to the monomer.

[0094] By performing chain transfer polymerization of a monomer having an alicyclic group in the presence of a compound having a crosslinkable silyl group and a chain-transfer functional group, a polymer represented by formula (IV), i.e., a polymer having a crosslinkable silyl group and an alicyclic group, can be obtained. This polymer has a crosslinkable silyl group at its terminal, as shown in formula (IV).

[0095]

[0096] In formula (IV), T' represents a group derived from T in formula (III). In other words, T' is a residue of T produced by chain transfer polymerization. For example, if T is a 12-mercaptododecyl group, then -T'-Y-SiR ３－ｎ Z ｎ is, -SC １２ H ２４ -Y-SiR ３－ｎ Z ｎ That is the case.

[0097] In formula (IV), Y, R, Z, and n are the same as Y, R, Z, and n in formula (III), respectively, and the preferred embodiment is the same.

[0098] In formula (IV), R １ and R ２ These are R in equation (1), respectively. １ and R ２ This is synonymous with the same thing, and the preferred form is also the same.

[0099] In formula (IV), q represents a value between 10 and 3,000. Preferably, q is between 15 and 2,500, and more preferably between 20 and 2,000.

[0100] A silane coupling reaction can be used as a method for bonding the crosslinkable silyl group of a polymer having a crosslinkable silyl group and an amine-based side chain to a functional group on the surface of a support. Known silane coupling reactions may be used, or any reaction similar thereto may be employed.

[0101] Formula (V) shows a structure that is presumed to be a preferred embodiment of the stationary phase obtained by supporting a polymer having amine-based side chains on a support in step (b).

[0102]

[0103] In equation (V), Z' and V are equivalent to Z' and V in equation (II), respectively.

[0104] In formula (V), Y, T', R １ , R ２ , and q are Y, T', and R in equation (IV), respectively. １ , R ２ , and are synonymous with q, and the preferred form is the same.

[0105] The following describes step (c). A preferred silane coupling agent having polymerizable functional groups used in step (c) is, for example, the compound represented by formula (I). The preferred embodiment of the compound represented by formula (I) is the same as that used in step (a) when used in step (c).

[0106] The monomer having an alicyclic group used in step (c) is the same monomer having an alicyclic group used in step (a), that is, the monomer represented by formula (2).

[0107] The polymerization method for a monomer having an alicyclic group and a silane coupling agent having a polymerizable functional group is not particularly limited, and any polymerization method can be employed, but radical polymerization is preferred, and living radical polymerization is more preferred. Radical polymerization is preferably carried out in the presence of a catalytic amount of radical generator. Any known radical generator or similar radical generator can be used. Examples of known radical generators include azo compounds and peroxides. Furthermore, polymerization is preferably carried out in the presence of a chain transfer agent, as this facilitates the control of the molecular weight of the polymer having amine side chains.

[0108] As a method for bonding the obtained polymer (i.e., a polymer having an alicyclic group) to the support by a silane coupling reaction, any known silane coupling reaction or a similar reaction can be used.

[0109] A preferred configuration of the stationary phase obtained by supporting a polymer having an alicyclic group on a support in step (c) is shown in formula (II).

[0110] The following describes step (d). A preferred silane coupling agent having polymerizable functional groups used in step (d) is, for example, the compound represented by formula (I). The preferred embodiment of the compound represented by formula (I) is the same as that used in step (a) when used in step (d).

[0111] The monomer having an alicyclic group used in step (d) is the same as the monomer having an alicyclic group used in step (a), that is, the monomer represented by formula (2).

[0112] When polymerizing a monomer having an alicyclic group and a silane coupling agent having a polymerizable functional group in the presence of a support, the polymerization method is not particularly limited, and any polymerization method can be employed, but free radical polymerization is preferred. Free radical polymerization is preferably carried out in the presence of a catalytic amount of a radical generator. Any known radical generator or similar radical generator can be used. Known radical generators include azo compounds and peroxides.

[0113] A preferred configuration of the stationary phase obtained by supporting a polymer having an alicyclic group on a support in step (d) is shown in formula (II).

[0114] The following describes step (e). The chain-transfer functional group introduced (chemically bonded) to the surface of the support used in step (e) can be the same type of group as the chain-transfer functional group of the compound having a crosslinkable silyl group and a chain-transfer functional group used in step (b). Therefore, as a support on which a chain-transfer functional group is introduced to the surface, for example, a compound represented by formula (III) bonded to the support by a silane coupling reaction is preferred.

[0115] The monomer having an alicyclic group used in step (e) is the same monomer having an alicyclic group used in step (a), that is, the monomer represented by formula (2).

[0116] Chain transfer polymerization is preferably carried out in the presence of a catalytic amount of radical generator. Any known radical generator or similar radical generator can be used. Examples of known radical generators include azo compounds and peroxides. In this case, the molecular weight can be controlled to some extent by the molar ratio of the chain transfer agent to the monomer.

[0117] A preferred configuration of the stationary phase obtained by supporting a polymer having an alicyclic group on a support in step (e) is shown in formula (V).

[0118] The following describes step (f). The atom transfer radical polymerization initiator group used in step (f) of the support to which the atom transfer radical polymerization initiator group has been introduced is preferably an α-haloalkylcarbonyl group. Examples of α-haloalkylcarbonyl groups include α-bromoisobutyryl group, α-bromopropionyl group, bromoacetyl group, and chloroacetyl group, with α-bromoisobutyryl group being preferred.

[0119] When silica particles are used as a support, a "support to which atom transfer radical polymerization initiator groups have been introduced" can be used if atom transfer radical polymerization initiator groups are chemically bonded to the silica particles via the silanol groups present in the silica particles.

[0120] When using a support other than silica particles, surface treatment of the support can suppress excessive adsorption of the sample onto the support itself, and also allow chemical bonding with atomic transfer radical polymerization initiator groups via groups introduced by the surface treatment. Examples of surface treatment agents include silane coupling agents such as 3-aminopropyltrimethoxysilane; titanium coupling agents; and aluminate coupling agents.

[0121] A support to which an atom transfer radical polymerization initiator group is attached can be obtained, for example, by a silane coupling reaction between a compound represented by formula (VI) and a support, preferably silica particles. A-Q-Y-SiR ３－ｎ Z ｎ (VI)

[0122] In formula (VI), A represents an atom transfer radical polymerization initiator group. The atom transfer radical polymerization initiator group is preferably an α-haloalkylcarbonyl group, more preferably an α-bromoisobutyryl group, an α-bromopropionyl group, a bromoacetyl group, or a chloroacetyl group, and even more preferably an α-bromoisobutyryl group.

[0123] In formula (VI), Q is an oxygen atom, NH, or NCH. ３ It represents, preferably NH.

[0124] In formula (VI), Y, R, Z, and n are the same as Y, R, Z, and n in formula (I), respectively, and the same applies to the preferred embodiments.

[0125] The compound represented by formula (VI) may be a commercially available compound, but it may also be a compound containing A and -Y-SiR ３－ｎ Z ｎ It can also be produced by reacting it with a compound having a group represented by . Furthermore, the reaction of these compounds with each other produces the "-X-" in formula (VI).

[0126] Compounds having the structure represented by A include compounds in which a halogen atom such as a chlorine atom, a bromine atom, and an iodine atom is bonded to an atom transfer radical polymerization initiator group. Examples of such compounds include α-bromoisobutyryl bromide, α-bromoisobutyryl chloride, α-bromopropionyl bromide, α-bromopropionyl chloride, bromoacetyl bromide, bromoacetyl chloride, chloroacetyl bromide, and chloroacetyl bromide chloride, with α-bromoisobutyryl bromide being preferred.

[0127] -Y-SiR ３－ｎ Z ｎ Examples of compounds having the group represented by include silane coupling agents having an alkoxy group with 1 to 5 carbon atoms as Z.

[0128] The support to which the atom transfer radical polymerization initiator group is attached is preferably a surface-modified silica particle obtained by a silane coupling reaction between a compound represented by formula (VI) and silica particles.

[0129] In step (f), when a polymer having an alicyclic group is supported on a carrier, the polymer having an alicyclic group is produced by atom transfer radical polymerization of a monomer having an alicyclic group, using an atom transfer radical polymerization initiator group bonded to the carrier as the initiator. Any conditions can be used as reaction conditions for atom transfer radical polymerization, such as conditions known in the field of polymer synthesis and conditions similar thereto.

[0130] The monomer having an alicyclic group used in step (f) is the same monomer having an alicyclic group used in step (a), that is, the monomer represented by formula (2).

[0131] Formula (VII) shows a structure that is presumed to be a preferred embodiment of the stationary phase obtained by supporting a polymer having an alicyclic group on a support in step (f).

[0132]

[0133] In formula (VII), A' represents a group derived from A in formula (VI). In other words, A' represents a residue of A generated by atom transfer radical polymerization. For example, if A is an α-bromoisobutyryl group, then -A'-Q-Y-SiR ３－ｎ Z ｎ is, -(CH ３ ) ２ CCO-Q-Y-SiR ３－ｎ Z ｎ That is the case.

[0134] In equation (VII), Z' and V are equivalent to Z' and V in equation (II), respectively.

[0135] In formula (VII), Y and Q are the same as Y and Q in formula (VI), respectively, and the preferred embodiments are the same.

[0136] In formula (VII), R １ and R ２ These are R in equation (1), respectively. １ and R ２ This is synonymous with the same thing, and the preferred form is also the same.

[0137] In formula (VII), q represents a value between 10 and 3,000. Preferably, q is between 15 and 2,500, and more preferably between 20 and 2,000.

[0138] The following describes step (g). Examples of compounds having a crosslinkable silyl group and an atom transfer radical polymerization initiator group used in step (g) include the compound represented by formula (VI).

[0139] The monomer having an alicyclic group used in step (g) is the same monomer having an alicyclic group used in step (a), that is, the monomer represented by formula (2).

[0140] By atom transfer radical polymerization of a monomer having an alicyclic group in the presence of a compound having a crosslinkable silyl group and an atom transfer radical polymerization initiator group, a polymer represented by formula (VIII), i.e., a polymer having a crosslinkable silyl group and an amine-based side chain, can be obtained. This polymer has a crosslinkable silyl group at its terminal, as shown in formula (VIII).

[0141]

[0142] In formula (VIII), Q, Y, R, Z, and n are the same as Q, Y, R, Z, and n in formula (VI), respectively, and the preferred embodiment is the same.

[0143] In equation (VIII), A' is equivalent to A' in equation (VII).

[0144] In formula (VIII), R １ and R ２ These are R in equation (1), respectively. １ and R ２ This is synonymous with the same thing, and the preferred form is also the same.

[0145] In formula (VIII), q represents a value between 10 and 3,000. Preferably, q is between 15 and 2,500, and more preferably between 20 and 2,000.

[0146] A silane coupling reaction can be used as a method for bonding the crosslinkable silyl groups of a polymer having crosslinkable silyl groups and alicyclic groups to the functional groups on the surface of the support. Known silane coupling reactions may be used, or any reaction similar thereto may be employed.

[0147] Formula (IX) shows a structure that is presumed to be a preferred embodiment of the stationary phase obtained by supporting a polymer having an alicyclic group on a carrier in step (g).

[0148]

[0149] In equation (IX), Z' and V are equivalent to Z' and V in equation (II), respectively.

[0150] In formula (IX), Y, Q, A', R １ , R ２ , and q are Y, Q, A', and R in equation (VIII), respectively. １ , R ２ , and are synonymous with q, and the preferred form is the same.

[0151] Step (h) is described below. Any method can be used to coat the carrier surface with the polymer having an alicyclic group, such as known methods and methods similar thereto. A known method is preferably one in which a coating solution containing the polymer having an alicyclic group and a solvent is coated (physically adsorbed) onto the carrier surface, and then the solvent is removed. The solvent in the coating solution is not particularly limited as long as it can dissolve the polymer having an alicyclic group, but examples include water; aprotic polar solvents such as N,N-dimethylformamide (DMF), N,N-dimethylacetamide (DMAc), dimethyl sulfoxide (DMSO), and N-methyl-2-pyrrolidone (NMP); cyclic amines such as pyridine; cyclic ethers such as THF and 1,4-dioxane; alcohols such as methanol and ethanol; and mixtures of two or more solvents selected from these. Of these, the solvent of the coating solution is preferably water, an aprotic polar solvent, a cyclic amine, a cyclic ether, or a mixture of two or more solvents selected from these, in terms of higher solubility of polymers having alicyclic groups; more preferably water, an aprotic polar solvent, or a mixture of two or more solvents selected from these; and even more preferably water, DMF, or a mixture thereof.

[0152] Polymers having alicyclic groups can be produced by a polymerization step in which monomers represented by formula (2) are polymerized. If the polymer having alicyclic groups contains other structural units, the polymerization step may involve polymerizing monomers that further produce other structural units together with the monomers having alicyclic groups. Examples of monomers that produce other structural units include any polymerizable compounds such as styrene compounds and olefin compounds.

[0153] The polymerization method for monomers having alicyclic groups is not particularly limited, and any polymerization method such as atom transfer radical polymerization, free radical polymerization, and chain transfer polymerization can be employed. Any reaction conditions for polymerization can be adopted, including conditions known in the field of polymer synthesis and conditions similar thereto.

[0154] 1-4. Chromatography The chromatography to which the stationary phase according to this embodiment is applied is not particularly limited, but liquid chromatography and supercritical fluid chromatography are preferred.

[0155] Examples of liquid chromatography include, but are not limited to, normal-phase chromatography, reverse-phase chromatography, size exclusion chromatography, thin-layer chromatography, and ion exchange chromatography. The stationary phase according to this embodiment can be suitably used as a stationary phase for reverse-phase chromatography because the polymer having alicyclic groups can provide hydrophobic interactions.

[0156] 2. Method for producing a stationary phase for chromatography A second embodiment of the present disclosure is a method for producing a stationary phase for chromatography, comprising any of the steps (a) to (h). The monomer having an alicyclic group in (a) to (g) is a monomer represented by formula (2). The polymer having an alicyclic group in (h) has a constituent unit represented by formula (1).

[0157] Steps (a) to (h) in this embodiment are the same as steps (a) to (h) described in the description of the first embodiment, and the preferred embodiments are also the same. Furthermore, the components represented by formula (1), the monomers represented by formula (2), and the carriers are the same as the components represented by formula (1), the monomers represented by formula (2), and the carriers in the first embodiment, and the preferred embodiments are also the same.

[0158] The manufacturing method according to this embodiment makes it possible to produce the chromatographic stationary phase according to the first embodiment; however, the chromatographic stationary phase produced by the manufacturing method according to this embodiment is not limited to the chromatographic stationary phase according to the first embodiment.

[0159] The manufacturing method according to this embodiment may include any steps other than those described above. Examples of optional steps include the oxidation step and the crosslinking step described in the description of the first embodiment.

[0160] 3. Separation Method A third embodiment of this disclosure is a separation method that includes a separation step of separating a sample by chromatography using the chromatography stationary phase according to the first embodiment. Examples of chromatography include liquid chromatography and supercritical fluid chromatography, with liquid chromatography being preferred. In the stationary phase according to the first embodiment, a polymer having an alicyclic group can provide hydrophobic interactions, so among liquid chromatography, reversed-phase chromatography is particularly preferred.

[0161] Chromatography can be performed using commercially available chromatographs, such as liquid chromatographs and supercritical fluid chromatographs. Column equilibration conditions and flow rates can be selected according to the column size, sample volume, and mobile phase type.

[0162] The sample to be separated by the separation method according to this embodiment is not particularly limited, but it is preferable that it contains hydrophobic substances. The stationary phase according to the first embodiment can provide hydrophobic interactions due to polymers having alicyclic groups. As a result, the stationary phase according to the first embodiment can strongly retain hydrophobic substances, and therefore the separation method according to this embodiment is considered suitable for separating these substances. Examples of hydrophobic substances include aromatic hydrocarbons and aliphatic hydrocarbons. Furthermore, as shown in the examples described later, the stationary phase according to the first embodiment has high molecular structure selectivity, so the separation method according to this embodiment can effectively separate positional isomers and structural isomers such as cis-trans isomers.

[0163] The solvent for the mobile phase when performing separation by liquid chromatography is not particularly limited, but it is preferably an organic solvent that can dissolve the target to be separated. Examples include organic solvents, water, and mixed solvents of water and organic solvents.

[0164] The organic solvent is not particularly limited, but examples include acetonitrile, methanol, ethanol, n-propanol, 2-propanol, tetrahydrofuran, 2-methyltetrahydrofuran, dimethyl sulfoxide (DMSO), N,N-dimethylformamide (DMF), and dimethylacetamide (DMAc), acetone, methyl ethyl ketone, diethyl ketone, methyl tert-butyl ether, chloroform, and dichloromethane. The organic solvent may also be a mixture of hydrocarbons having 5 to 8 carbon atoms and alcohols, which are commonly used in chromatography. The organic solvent may be used alone, or two or more may be used in any combination and ratio.

[0165] Of these, the organic solvent is preferably one or more selected from acetonitrile, methanol, ethanol, n-propanol, 2-propanol, and tetrahydrofuran, in terms of high chemical stability, more preferably one or more selected from methanol and acetonitrile, and even more preferably methanol.

[0166] Examples of solvents for the mobile phase when performing separation by supercritical fluid chromatography include supercritical carbon dioxide, a mixture of supercritical carbon dioxide and an organic solvent, and a mixture of supercritical carbon dioxide, an organic solvent, and water. The organic solvents included in these mixtures are preferably those described as organic solvents used in the mobile phase for liquid chromatography.

[0167] The present disclosure will be further described below with reference to examples, but the present disclosure is not limited to the following examples without departing from its essence. In this disclosure, "room temperature" means 25°C. A Yanaco CHN coder MT-5 was used for elemental analysis.

[0168] [Synthesis Example 1: Synthesis of Silica Particles with Atomic Transfer Radical Polymerization Initiator Groups] 6.645 g (30 mmol) of 3-aminopropyltriethoxysilane and 5 mL (36 mmol) of triethylamine were dissolved in 20 mL of tetrahydrofuran to obtain a solution, which was stirred in an ice bath. 4.45 mL (36 mmol) of 2-bromoisobutyryl bromide was added dropwise, the reaction mixture was heated to room temperature, and stirred overnight under an argon atmosphere. After the reaction, the reaction mixture was concentrated in an evaporator to obtain 12.075 g (76.8% yield) of 3-(α-bromoisobutylamide)propyltriethoxysilane. １ The H-NMR spectral data is shown below.

[0169] 1 H-NMR (300 MHz, CDCl3, δ / ppm): 3.82-3.74(6H, 4 cracks, SiOCH2CH3), 3.27(2H, 4 cracks, SiCH2CH2CH2NH), 1.92(6H, s, SiCH2CH2CH2NH), 1.65(2H, 5, SiCH2CH2CH2NH), 1.24(9H, t, SiCH2CH3), 0.62(6H, t, SiCH2CH2CH2NH)

[0170] Porous silica particles ("ChromatoRex SPS100-5" manufactured by Fuji Silicia Chemical Co., Ltd., particle size 5 μm, specific surface area 300 m²) ２ 10.215 g of (1 / g, average pore size 10 nm), 7.516 g (14 mmol) of 3-(α-bromoisobutylamide)propyltriethoxysilane, and 1 mL of pyridine were mixed, and the resulting reaction solution was heated under reflux for 1 day.

[0171] After the reaction, the precipitate in the reaction solution was filtered, washed with methanol, and dried to obtain 15.467 g of silica particles into which atom transfer radical polymerization initiator groups were introduced. The silica particles into which atom transfer radical polymerization initiator groups were introduced (see below; hereinafter referred to as "ATRPini-silica") are presumed to have the following structure.

[0172]

[0173] [Synthesis Example 2: Synthesis of Menthyl Methacrylate] 6.317 g (40.4 mmol) of L-menthol was dissolved in 15 mL of dry toluene, and then mixed with 5.214 g (49.9 mmol) of methacryloyl chloride to obtain an L-menthol solution. While cooling the L-menthol solution, 7 mL (50 mmol) of triethylamine was added dropwise to the solution, and after the heat of reaction subsided, the mixture was stirred overnight at room temperature.

[0174] The product was extracted from the reaction mixture with ethyl acetate, and the extract was sequentially washed with dilute hydrochloric acid, aqueous sodium carbonate solution, and saline solution. After drying the extract over magnesium sulfate, it was concentrated using an evaporator to obtain 7,600 g of menthyl methacrylate (yield 88%). １ The H-NMR spectral data is shown below.

[0175] 1 H-NMR (300 MHz, CDCl3, δ / ppm): 6.08(1H,s), 5.52(1H, s), 4.73(1H, 6-fold), 2.51(2H, q), 2.29(3H, s), 1.91(6.6-4, m), 1.67(3H, m), 1.52-1.44(3H, m), 1.11-0.88(12H, m), 0.76(4H, d)

[0176] [Synthesis Example 3: Synthesis of Bornyl Methacrylate] 3.107 g (20 mmol) of borneol was dissolved in 15 mL of dry toluene, and then mixed with 3.180 g (30 mmol) of methacryloyl chloride to obtain a borneol solution. While cooling the borneol solution, 4.2 mL (30 mmol) of triethylamine was added dropwise to the solution, and the mixture was stirred overnight at room temperature.

[0177] The product was extracted from the reaction mixture with ethyl acetate, and the extract was sequentially washed with dilute hydrochloric acid, aqueous sodium carbonate solution, and saline solution. After drying the extract over magnesium sulfate, it was concentrated in an evaporator and dried under reduced pressure to obtain 2.605 g of bornyl methacrylate (yield 58.6%). １ The H-NMR spectral data is shown below.

[0178] 1 H-NMR (300 MHz, CDCl3, δ / ppm): 6.11(1H, s, 2HC=CCH3), 5.55(1H, s, 2HC=CCH3), 4.92(1H, 8), 2.35(4H, s), 1.96(4H, t), 1.70(3H, m), 1.29(2H, m), 0.93(3H, s), 0.89(3H, s), 0.86(3H, s)

[0179] [Example 1: Preparation of stationary phase for chromatography]

[0180] 0.841 g of ATRPini-silica, 0.581 g (2.5 mmol) of menthyl methacrylate, 70 mg (0.031 mmol) of copper(II) bromide, and 0.108 g (0.62 mmol) of N,N,N',N'',N''-pentamethyldiethylenetriamine were dissolved in 7 mL of N,N-dimethylformamide to obtain a monomer solution. 0.259 g (1.5 mmol) of ascorbic acid was added to this monomer solution, and the resulting reaction mixture was stirred overnight at 60°C.

[0181] After the reaction, the reaction solution was mixed with chloroform, and the solid was recovered by centrifugation (1,500 rpm, 10 min). This solid was mixed with an aqueous solution of ethylenediaminetetraacetic acid, and the solid was centrifuged under the same conditions as above to obtain 1.031 g of a stationary phase in which poly(menthyl methacrylate) was supported on silica particles. Elemental analysis of the obtained stationary phase was performed, and the degree of organic modification was calculated to be 26.91% by mass.

[0182] The obtained stationary phase was dispersed in 10 mL of a mixed solvent of isopropanol and chloroform (50 / 50 (v / v)), and packed into a stainless steel column (3 mm inner diameter, 100 mm length) to prepare a chromatography column.

[0183] [Example 2: Preparation of stationary phase for chromatography] 0.759 g of ATRPini-silica, 5 mL of N,N-dimethylformamide, 33 mg of copper(II) bromide, 0.194 g of N,N,N',N'',N''-pentamethyldiethylenetriamine, and 0.382 g (1.5 mmol) of bornyl methacrylate were mixed, and the resulting mixture was degassed by ultrasonic irradiation and reduced pressure. 0.220 g of L-ascorbic acid was added to this mixture, and the resulting reaction solution was stirred at 60°C for 24 hours.

[0184] After the reaction, the reaction mixture was mixed with methanol, and a dispersion was obtained by irradiating the resulting mixture with ultrasound. The solid in this dispersion was recovered by centrifugation (1,500 rpm, 10 min). This solid was mixed with N,N-dimethylform, and the centrifugation of the solid was repeated twice under the same conditions as above. Subsequently, this solid was mixed with an aqueous solution of ethylenediaminetetraacetic acid, and the centrifugation of the solid was repeated twice under the same conditions as above to obtain 0.844 g of a stationary phase in which poly(bornyl methacrylate) was supported on silica particles. Elemental analysis of the obtained stationary phase was performed, and the degree of organic modification was calculated to be 25.46% by mass.

[0185] A chromatography column was prepared in the same manner as in Example 1, except that the stationary phase obtained in this example was used.

[0186] [Example 3: Preparation of stationary phase for chromatography] 0.789 g of ATRPini-silica, 5 mL of N,N-dimethylformamide, 43 mg of copper(II) bromide, 0.193 g of N,N,N',N'',N''-pentamethyldiethylenetriamine, and 0.332 g (1.5 mmol) of 1-adamantyl methacrylate were mixed, and the resulting mixture was degassed by ultrasonic irradiation and reduced pressure. 0.247 g of L-ascorbic acid was added to this mixture, and the resulting reaction solution was stirred at 60°C for 24 hours.

[0187] After the reaction, the reaction solution was mixed with methanol, and a dispersion was obtained by irradiating the resulting mixture with ultrasound. The solid in this dispersion was recovered by centrifugation (1,500 rpm, 10 min). This solid was mixed with N,N-dimethylform, and the centrifugation procedure was repeated twice under the same conditions as above. Subsequently, this solid was mixed with an aqueous solution of ethylenediaminetetraacetic acid, and the centrifugation procedure was repeated twice under the same conditions as above to obtain 0.877 g of a stationary phase in which poly(1-adamantyl methacrylate) was supported on silica particles. Elemental analysis of the obtained stationary phase was performed, and the degree of organic modification was calculated to be 29.42% by mass.

[0188] A chromatography column was prepared in the same manner as in Example 1, except that the stationary phase obtained in this example was used.

[0189] [Comparative Example 1: Stationary Phase for Chromatography] A commercially available column (Merck "Prospher RP-18 Endcapped", particle size 5 μm, inner diameter 4.6 mm, length 150 mm) was used as a chromatography column packed with the chromatographic stationary phase of Comparative Example 1. Elemental analysis of the stationary phase in this column was performed, and the degree of organic modification was calculated to be 23.42% by mass. The chromatographic stationary phase of Comparative Example 1 is a stationary phase in which octadecylsilyl groups are monomerically chemically bonded to the surface of silica particles (see below).

[0190]

[0191] [Comparative Example 2: Stationary Phase for Chromatography] A commercially available column ("CAPCELL PAK ADME-HR" manufactured by Osaka Soda Co., Ltd., particle size 3 μm, inner diameter 4.6 mm, length 100 mm) was used as a chromatography column packed with the chromatographic stationary phase of Comparative Example 2. The chromatographic stationary phase of Comparative Example 2 is a stationary phase in which 2-(1-adamantyl)ethylsilyl groups are monomerically chemically bonded to the surface of silica particles (see below).

[0192]

[0193] [Comparative Example 3: Stationary Phase for Chromatography] Porous silica particles ("ChromatoRex SPS100-5" manufactured by Fuji Silicia Chemical Co., Ltd., particle size 5 μm, specific surface area 300 m²) ２ 0.800 g of porous silica particles (average pore size 10 nm) was dried overnight at 150°C. The dried porous silica particles were mixed with 5 mL of dimethoxyethane, 0.79 g (2.04 mmol) of trichlorooctadecylsilane, and 0.5 mL of pyridine, and the resulting reaction solution was heated under reflux for 1 day.

[0194] After the reaction, methanol was added to the reaction mixture, and the solid was filtered off. This solid was dispersed in acetone to obtain a dispersion. The solid in this dispersion was recovered by centrifugation (1,500 rpm, 10 min) to obtain a stationary phase in which silica particles were modified with octadecylsilyl groups. This stationary phase is one in which octadecylsilyl groups are polymerically chemically bonded to the surface of silica particles (see below).

[0195]

[0196] R ｘ These are groups (e.g., hydroxyl groups or octadecyl groups) that are generated during the process of stationary phase production.

[0197] [Comparative Example 4: Stationary Phase for Chromatography] A commercially available column (YMC-Pack Pro C18, manufactured by YMC Corporation, particle size 6 μm, inner diameter 4.6 mm, length 150 mm) was used as a chromatography column packed with the chromatographic stationary phase of Comparative Example 4. The chromatographic stationary phase of Comparative Example 4 is a stationary phase in which octadecylsilyl groups are polymerically chemically bonded to the surface of methacrylate polymer particles.

[0198] [Stationary Phase Evaluation I: Tanaka Test] Reverse-phase high-performance liquid chromatography (reverse-phase HPLC) was performed using the chromatography columns of the examples and comparative examples at the flow rates and back pressures shown in Table 1, and the stationary phase was evaluated by the Tanaka test (see N. Tanaka, J. Chromatogr., 239, 761-772 (1982)). In reverse-phase HPLC, a mixture of methanol and water (80 / 20 (v / v)) was used as the mobile phase. The retention coefficient and selectivity obtained based on the following formula are shown in Table 2.

[0199] • The retention coefficient of substance X is k(X) = (t Ｒ -t ０ ) / t ０ t Ｒ : Retention time t of substance X ０ : Elution time of non-retained substances

[0200] Selectivity of substance Y for substance Z α(Y / Z) = k(Y) / k(Z) k(Y): retention coefficient of substance Y k(Z): retention coefficient of substance Z

[0201] [Stationary Phase Evaluation II: Neue Test] Reverse-phase HPLC was performed using the chromatography columns of the examples and comparative examples at the flow rates and back pressures shown in Table 3, and the stationary phase was evaluated by the Neue test (see SAWise, WJBonnett, FRGuenther, WEMay, J. Chromatogr. Sci., 19, 457 (1981)). In reverse-phase HPLC, a mixture of methanol and 20 mM potassium phosphate buffer with a pH of 7.0 at 25°C (65 / 35 (v / v)) was used as the mobile phase. The retention coefficient and selectivity obtained based on the formula described in the above section [Stationary Phase Evaluation I: Tanaka Test] are shown in Table 4.

[0202]

[0203]

[0204]

[0205]

[0206] As can be seen from Table 2, the stationary phase prepared in Example 1 has a higher retention coefficient for amylbenzene than for butylbenzene, and a higher retention coefficient for triphenylene than for o-terphenyl. From this, it can be seen that the stationary phase prepared in Example 1 is a stationary phase that strongly retains highly hydrophobic compounds. Furthermore, from Table 2, the stationary phase prepared in Example 1 had a very high selectivity (α(T / O)) of 2.28 for triphenylene relative to o-terphenyl. Since it is rare for conventional stationary phases to have an α(T / O) value greater than 2.0, the high α(T / O) can be said to be a characteristic attributable to the polymer containing an alicyclic group.

[0207] Table 4 shows that the stationary phase prepared in Example 1 exhibits slightly higher selectivity for butylparaben relative to dipropyl phthalate in the Neue test. Furthermore, the stationary phase prepared in Example 1 shows high affinity and strong retention for naphthalene, acenaphthalene, butylparaben, dipropyl phthalate, and amitriptyline.

[0208] As can be seen from Table 2, the stationary phase prepared in Example 3 had a retention coefficient for o-terphenyl greater than that for triphenylene, resulting in a selectivity of 0.901 for triphenylene relative to o-terphenyl, which is less than 1. In conventional stationary phases such as those prepared in Comparative Examples 1 and 2, the planar compound triphenylene (logP = 5.729) is generally retained more strongly than the non-planar compound o-terphenyl (logP = 5.167), which is less hydrophobic. On the other hand, the stationary phase prepared in Example 3 retained o-terphenyl more strongly than triphenylene, confirming that it exhibits the opposite selectivity to conventional stationary phases.

[0209] "logP" is the water / 1-octanol partition coefficient and is an indicator of whether an object is hydrophobic or hydrophilic. The more hydrophobic the object, the larger the logP value.

[0210] As can be seen from Table 2, the stationary phase prepared in Example 3 has low retention coefficients for o-terphenyl and triphenylene. In particular, among the examples and comparative examples, only Example 3 had a stationary phase in which the retention coefficients for o-terphenyl and triphenylene were lower than those for butylbenzene. From these results, it can be inferred that the stationary phase prepared in Example 3 has weak retention of molecules with similar shapes to o-terphenyl and triphenylene.

[0211] Diisopropyl phthalate (logP = 3.733) is more hydrophobic than butylparaben (logP = 3.41). Therefore, in conventional stationary phases such as those prepared in Comparative Examples 1 and 2, the retention coefficient of butylparaben is generally smaller than that of dipropyl phthalate. On the other hand, as can be seen from Table 4, in the Neue test, the chromatographic stationary phase prepared in Example 3 showed a higher retention coefficient of butylparaben than that of dipropyl phthalate, indicating very high selectivity for butylparaben over diisopropyl phthalate. This confirms that the stationary phase prepared in Example 3 exhibits selectivity opposite to that of conventional stationary phases.

[0212] Table 3 shows that the stationary phase prepared in Example 3 exhibits a very high selectivity of naphthalene for acenaphthene, at 0.91.

[0213] From the above, it was found that among the stationary phases described above, the stationary phase prepared in Example 3 has special retention characteristics that differ from those of conventional stationary phases.

[0214] Both o-terphenyl and dipropyl phthalate are ortho isomers. From stationary phase evaluations I and II, it was inferred that the retention of disubstituted benzenes was weaker because the two substituents being in the ortho position made the molecules bulkier. Therefore, the following evaluations were performed.

[0215] [Stationary Phase Evaluation III: Tanaka Test] The Tanaka test was performed in the same manner as in Stationary Phase Evaluation I. The literature values ​​(refer to SciFinder) for retention coefficient, selectivity, and logP are shown in Table 5. Figures 1 to 5 show graphs plotting the data for the stationary phases of Example 3 and Comparative Examples 1 to 4, with logP on the vertical axis and logk on the horizontal axis.

[0216]

[0217]

[0218] The annotations in Tables 5 and 6 are explained below: α(m / A) = α(metha isomer / amylbenzene) α(m / o) = α(metha isomer / ortho isomer) α(p / o) = α(para isomer / ortho isomer) α(Pr / isoPr) = α(dipropyl phthalate / diisopropyl phthalate) α(c / A) = α(cis-stilbene / amylbenzene) α(t / c) = α(trans-stilbene / cis-stilbene) α(TPE / c) = α(tetraphenylethylene / cis-stilbene)

[0219] As can be seen from Tables 5 and 6, in the stationary phases prepared in Comparative Examples 1 to 4, bis(propoxymethyl)benzene had the largest retention coefficient for the meta-isomer, and the selectivity for the meta-isomer and para-isomer relative to the ortho-isomer was 1 or less. In contrast, in the stationary phase prepared in Example 3, the retention coefficient for the para-isomer was the largest, and the retention coefficient for the ortho-isomer tended to be small. From these findings, it can be seen that the stationary phase prepared in Example 3 differs from conventional stationary phases in that it exhibits high para-isomer selectivity.

[0220] Furthermore, Tables 5 and 6 show that the stationary phase prepared in Example 3 tends to have higher selectivity not only for para-isomers to ortho-isomers, but also for meta-isomers to ortho-isomers, compared to the stationary phases prepared in Comparative Examples 1 to 4. Among the disubstituted benzenes, particularly high meta-isomer selectivity and para-isomer selectivity were observed when dinitrobenzene, terphenyl, and dipropyl phthalate were used as samples. This is due to the low retention of ortho-isomers of disubstituted benzenes in the stationary phase prepared in Example 3.

[0221] In the stationary phase prepared in Example 3, despite tetraphenylethylene having a larger logP than cis-stilbene, the selectivity of tetraphenylethylene over cis-stilbene was low at 0.38. Since tetraphenylethylene has two more benzene rings per molecule than cis-stilbene, it should normally be retained more strongly by the stationary phase than cis-stilbene in reverse-phase chromatography; however, the stationary phase prepared in Example 3 showed the opposite result.

[0222] When examining the relationship between the retention coefficients of the dinitrobenzene, terphenyl, and dipropyl phthalate isomers and logP in the stationary phase prepared in Example 3, it was found that the retention coefficient of the ortho isomer was small relative to logP. Furthermore, when examining the relationship between the retention coefficients of the stilbene isomer and tetraphenylethylene and logP, it was found that the retention coefficient of tetraphenylethylene was significantly small relative to logP. In other words, although these compounds are highly hydrophobic, their retention coefficients are smaller than the values ​​that would be expected from their hydrophobicity. From the above, it is considered that the stationary phase prepared in Example 3 has large steric hindrance interactions with these compounds.

[0223] Next, FIGS. 1 to 5 will be considered. In reverse-phase HPLC, retention is mainly affected by the hydrophobicity of the sample, so it is known that logP and logk have a high correlation. In the graphs of FIGS. 1 to 5, the plots of alkylbenzenes were used as an index of hydrophobicity, and an approximate straight line was drawn for these plots. Compounds plotted below the approximate straight line can be said to have a large retention with respect to the hydrophobicity of the compound. On the other hand, compounds plotted above the approximate straight line can be said to have a small retention with respect to the hydrophobicity of the compound. The slope of these approximate straight lines is correlated with the selectivity α (CH ２ ), and the larger the slope, the smaller α (CH ２ ). The intercept of the equation of the approximate straight line represents "the logP required to obtain a retention coefficient of 1". The intercept of the equation of the approximate straight line is correlated with the retention coefficient of amylbenzene, and the larger the intercept, the smaller the retention coefficient of amylbenzene.

[0224] From FIGS. 1 to 5, it is observed that the structural isomers of 2-substituted benzenes tend to be plotted in the order of ortho isomers, meta isomers, and para isomers from the left side to the right side of the graph.

[0225] As shown in FIG. 1, the correlation coefficient of the approximate straight line of the plots of alkylbenzenes in the stationary phase prepared in Example 3 is 0.600, which is lower than that of the stationary phase prepared in the comparative example. From this, it is considered that the stationary phase prepared in Example 3 is greatly affected by factors other than hydrophobicity on retention. In particular, since the plots of dipropyl phthalate, diisopropyl phthalate, m-terphenyl, p-terphenyl, stilbene, and tetraphenyl ethylene deviate significantly from the approximate straight line, it is considered that the high steric hindrance of poly(1-adamantyl methacrylate) affects retention.

[0226] The stationary phase prepared in Comparative Example 4 had silica particles whose surfaces were modified with 2-(1-adamantyl)ethylsilyl groups. The correlation coefficient of the approximate straight line for the stationary phase prepared in Comparative Example 4 was 0.959, which was significantly higher than that of the stationary phase in Example 3, in which poly(1-adamantyl methacrylate) was supported on silica particles. From this, it can be said that in the stationary phase prepared in Comparative Example 4, the hydrophobicity of the sample had a dominant effect on retention, and the effect of steric hindrance from the adamantyl group on retention was much smaller compared to the stationary phase prepared in Example 3.

Claims

1. A stationary phase for chromatography comprising a carrier and a polymer having alicyclic groups supported on the carrier, wherein the carrier has a particle shape and the polymer having alicyclic groups has a structural unit represented by formula (1). (In the formula, R １ is an adamantyl group, a bornyl group, a norbornyl group, or a menthyl group; the adamantyl group, the bornyl group, the norbornyl group, and the menthyl group may each have one or more substituents selected from the group consisting of a deuterium atom, a halogen atom, a hydrocarbon group having 1 to 3 carbon atoms, and a halogenated hydrocarbon group having 1 to 3 carbon atoms; R ２ (This is either a hydrogen atom or a methyl group.) 2. R １ The chromatography stationary phase according to claim 1, wherein the group is an adamantyl group.

3. The chromatography stationary phase according to claim 1, wherein the polymer having the alicyclic group is supported on the carrier via chemical bonds.

4. The stationary phase for chromatography according to claim 1, wherein the carrier is porous particles or non-porous particles.

5. The chromatography stationary phase according to claim 4, wherein the constituent material of the carrier is one or more selected from the group consisting of silica, alumina, zirconia, titania, magnesia, glass, kaolin, silicate, and hydroxyapatite.

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

7. A separation method comprising a separation step of separating a sample by chromatography using a stationary phase for chromatography as described in any one of claims 1 to 6.

8. A method for producing a stationary phase for chromatography, comprising a supporting step of supporting a polymer having an alicyclic group on a carrier, wherein the supporting step is any of the following steps (a) to (h), the monomer having an alicyclic group in the following steps (a) to (g) includes a monomer represented by formula (2), and the polymer having an alicyclic group in the following step (h) has a constituent unit represented by formula (1). (a) A step of free radical polymerization of a monomer having an alicyclic group in the presence of a support into which a polymerizable functional group has been introduced. (b) A step of producing a crosslinkable polymer having the crosslinkable silyl group and the alicyclic group by chain transfer polymerization of the monomer having an alicyclic group in the presence of a compound having a crosslinkable silyl group and a chain transfer functional group, and reacting the crosslinkable silyl group of the crosslinkable polymer with the functional group on the surface of the support. (c) A step of producing a polymer-modified silane coupling agent by polymerizing a monomer having an alicyclic group and a silane coupling agent having a polymerizable functional group, and bonding the polymer-modified silane coupling agent to the support by a silane coupling reaction. (d) A step of polymerizing a monomer having an alicyclic group and a silane coupling agent having a polymerizable functional group in the presence of a support. (e) A step of chain transfer polymerization of a monomer having an alicyclic group in the presence of a support into which a chain transfer functional group has been introduced. (f) A step of atom transfer radical polymerization of a monomer having an alicyclic group in the presence of a support into which an atom transfer radical polymerization initiator group has been introduced. (g) A step of producing a crosslinkable polymer having the crosslinkable silyl group and the alicyclic group by atom transfer radical polymerization of a monomer having an alicyclic group in the presence of a compound having a crosslinkable silyl group and an atom transfer radical polymerization initiator group, and reacting the crosslinkable silyl group of the crosslinkable polymer with the functional group of the carrier surface. (h) A step of coating the carrier surface with the polymer having an alicyclic group. (In the formula, R １ is an adamantyl group, a bornyl group, a norbornyl group, or a menthyl group; the adamantyl group, the bornyl group, the norbornyl group, and the menthyl group may each have one or more substituents selected from the group consisting of a deuterium atom, a halogen atom, a hydrocarbon group having 1 to 3 carbon atoms, and a halogenated hydrocarbon group having 1 to 3 carbon atoms; R ２ (This is either a hydrogen atom or a methyl group.) 9. The method for producing a stationary phase for chromatography according to claim 8, wherein the loading step is step (f) or step (h).

10. The method for producing a stationary phase for chromatography according to claim 8, wherein the carrier is porous particles or non-porous particles.

11. The method for producing a stationary phase for chromatography according to claim 10, wherein the constituent material of the carrier is one or more selected from the group consisting of silica, alumina, zirconia, titania, magnesia, glass, kaolin, silicate, and hydroxyapatite.

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