JP5504421B2 - Column for liquid chromatography and method for producing the same - Google Patents

Description

  The present invention relates to a liquid chromatography column and a method for producing a liquid chromatography column.

  A general column for high performance liquid chromatography is produced by filling resin particles into an empty column (cylindrical tube). After resin particles are synthesized by suspension polymerization, etc., classification is necessary so that the particle size is uniform, and adjustment of packing pressure, packing time, etc. is necessary so that the column can be packed uniformly. It has become a big Kushiro.

  In order to solve this, a method of directly polymerizing raw materials in an empty column (tube) to form a porous resin (monolith) is known (in situ polymerization method) (for example, see Non-Patent Document 1). ). Monoliths are broadly classified into silica-based monoliths and organic polymer-based monoliths depending on the material, and silica monolithic columns are becoming widely used for applications such as protein separation (see Non-Patent Document 2, for example).

  On the other hand, the organic polymer monolith has characteristics such as higher alkali resistance than the silica monolith, but volumetric shrinkage tends to occur during polymerization in the column, and a gap is easily generated between the organic polymer monolith and the inner wall of the column. As a result, problems such as a monolith falling off (for example, see Patent Document 1) and a decrease in the number of theoretical plates are caused, so that it is rarely used as a column for general-purpose high performance liquid chromatography (for example, see Non-Patent Document 3).

  In order to reduce the gap between the organic polymer monolith and the inner wall of the column, a fused silica tube (manufactured by Polymicro Technologies Inc.) is used as an empty column (tube), and then a silane coupling agent having a double bond is reacted. A method is known in which an organic polymer monolith is produced by in situ polymerization, and the organic polymer monolith is fixed to the surface of a tube (for example, see Non-Patent Document 4).

  However, in this method, since a fused silica tube is used as an empty column, the alkali resistance is remarkably reduced, and the excellent pH resistance inherent in organic monoliths cannot be sufficiently exhibited, leading to a decrease in the number of theoretical plates. Remain.

Japanese Patent No. 3168006

Anal. Chem. 68, 3498 (1996) Chromolith Reversed-phase HPLC columns General Information and Guidelines, Merck Tomoya Umemura, Tokuhisa Kojima, Junji Ueki, “Development of high-speed and high-performance separation analysis method using organic polymer monolith”, Analytical Chemistry, Japan Analytical Chemistry Society, 2008, Vol. 57, no. 7, p. 517-529 Yi-Ming Li, et al. “Continuous Beds for Microchromatography: Cation-Exchange Chromatography”, Analytical Biochemistry, 223, 153-158 (1994).

  The present invention provides a liquid chromatography column that prevents a gap from occurring between the organic polymer monolith and the inner wall of the resin tube, prevents the organic polymer monolith from falling out of the resin tube during use, and provides a high theoretical plate number. It is for the purpose. The present invention also provides a liquid chromatography column that prevents a gap from being generated between the organic polymer monolith and the inner wall of the resin tube, prevents the organic polymer monolith from falling off from the resin tube during use, and provides a high theoretical plate number. The object is to provide a manufacturing method.

  The present inventors use a tube containing polyether ether ketone as a resin tube, and fix the resin tube and the organic polymer monolith by a chemical bond via a sulfonyl group, whereby the inner wall of the resin tube due to volume shrinkage during polymerization It has been found that separation of organic polymer monoliths (occurrence of gaps) from the resin can be suppressed, and a column for liquid chromatography having a high theoretical plate number in which organic polymer monoliths do not fall out of the resin tube during use has been invented.

That is, the present invention is a liquid chromatography column having a resin tube and an organic polymer monolith formed in the resin tube, wherein the resin contains a polyether ether ketone, and the organic polymer monolith is the resin The present invention relates to a liquid chromatography column which is bonded to a tube via a sulfonyl group.
As a preferred embodiment of the liquid chromatography column of the present invention, the organic polymer monolith is polymerized in the molecule by the liquid chromatography column in which the organic polymer monolith is bonded to the resin tube via a sulfonyloxy group. Column for liquid chromatography which is a monolith obtained by polymerizing a polymerizable monomer having a carbon-carbon double bond, and liquid chromatography wherein the organic polymer monolith is a (meth) acrylic resin monolith Column.
Further, the present invention introduces a vinyl group on the inner wall of the resin tube by introducing a sulfonic acid group into the inner wall of the resin tube and reacting the introduced sulfonic acid group with a vinyl monomer having a glycidyl group. Introducing and polymerizing a polymerizable monomer having a polymerizable carbon-carbon double bond in the molecule in the resin tube into which the vinyl group has been introduced, forms an organic polymer monolith in the resin tube. The present invention relates to a method for producing a column for liquid chromatography.

  According to the present invention, there is provided a liquid chromatography column that prevents a gap from being generated between the organic polymer monolith and the inner wall of the resin tube, prevents the organic polymer monolith from falling out of the resin tube during use, and provides a high theoretical plate number. Can be provided. In addition, according to the present invention, it is possible to prevent a gap from being generated between the organic polymer monolith and the inner wall of the resin tube, and the organic polymer monolith is difficult to fall out of the resin tube during use, so that a high theoretical plate number can be obtained. A method for producing a column can be provided.

It is the schematic which shows the sulfonyl group introduction | transduction process and vinyl group introduction | transduction process to the column made from a polyether ether ketone. It is the result of having analyzed the sample sample by the liquid chromatography using the column produced in Example 1. FIG. 2 is a cross-sectional SEM photograph of the liquid chromatography column obtained in Example 1. FIG. 2 is a cross-sectional SEM photograph of the liquid chromatography column obtained in Comparative Example 1.

Hereinafter, embodiments of the present invention will be described in detail.
The column for liquid chromatography (LC) of the present invention is characterized in that an organic polymer monolith is fixed to a resin tube by a bond that breaks a sulfonyl group.

  In the present invention, a tube formed of a resin containing polyether ether ketone (PEEK) is used as the resin tube. In addition to tubes formed of resin containing only PEEK as a resin component, PEEK and other resins such as polytetrafluoroethylene (PTFE), polyvinylidene fluoride (PVDF), polycarbonate, high density polypropylene, high density polyethylene, etc. It is also possible to use a tube formed of a mixed resin containing. Commercially available products can be used as PEEK tubes, such as NPK-026 and NPK-003 (manufactured by Nirei Kogyo Co., Ltd.).

  The resin that forms the resin tube does not contain PEEK, and a resin containing PTFE, PVDF, polycarbonate, high-density polypropylene, or high-density polyethylene may be used. PEEK is resistant to organic solvents. It is particularly suitable as an empty column (tube) for liquid chromatography because of its excellent acid resistance and alkali resistance, as well as excellent pressure resistance. PEEK is also preferable in that it is easy to introduce a sulfonyl group.

  Also, vinyl chloride without solvent resistance, steel without acid resistance, silica gel coating with poor alkali resistance, etc. are not suitable as an empty column (tube) for liquid chromatography.

  The diameter of the resin tube is not particularly limited, but if it is too large, there is a tendency that a gap is easily formed between the resin tube and the monolith due to thermal contraction, and the resin tube and the monolith are easily separated. On the other hand, if it is too small, the strength of the resin tube is insufficient, and it tends to bend or bend. Even if a monolith is formed, there is a tendency that the resin component is too small for liquid chromatography and separation is insufficient. . The diameter (inner diameter) of the resin tube can be set to, for example, 0.1 mm to 10 mm.

  The length of the resin tube is not particularly limited, and when a column for liquid chromatography is used, the longer one is advantageous because the number of theoretical plates can be easily obtained. However, if the resin tube is too long, it is easy to bend. Therefore, when the resin tube is long, it is desirable to fix it to a base or the like. The length of the resin tube can be, for example, 5 cm to 300 cm.

  Examples of the organic polymer monolith in the present invention include (meth) acrylic resin monoliths, styrene resin monoliths, and allyl resin monoliths. The (meth) acrylic resin monolith is preferably used from the viewpoint of ease of polymerization and ease of modification of the functional group. In the present invention, the organic polymer monolith can be formed, for example, by a polymerization reaction of a polymerizable monomer having a polymerizable carbon-carbon double bond (polymerizable group) in a molecule described later.

  In the liquid chromatography column of the present invention, an organic polymer monolith is bonded to a resin tube via a sulfonyl group. The organic polymer monolith bonded by a sulfonyl group is excellent in alkali resistance and acid resistance, and can exhibit the excellent pH resistance inherent in the organic polymer monolith.

In the liquid chromatography column of the present invention, for example, first, a sulfonic acid group is introduced into the inner wall of the resin tube, and then the introduced sulfonic acid group is reacted with a vinyl monomer having a glycidyl group. Thus, a vinyl group is introduced into the inner wall of the resin tube to form a scaffold for bonding the organic polymer monolith. Then, it can obtain by the method of superposing | polymerizing the polymerizable monomer used as the raw material of organic polymer monolith in the said resin tube into which the vinyl group was introduce | transduced.
Hereinafter, although this method is demonstrated in detail, the manufacturing method of the column for liquid chromatography of this invention is not limited to this method.

  First, a sulfonic acid group is introduced into a PEEK (polyether-ether-ketone) tube (sulfonic acid group introduction step), and then a vinyl monomer having a glycidyl group is reacted to introduce a vinyl group (vinyl group). The introduction process) process is described.

(Sulphonic acid group introduction step)
First, an aqueous sulfuric acid solution is injected into the PEEK tube, both ends are sealed, and reacted to introduce (sulfonate) a sulfonic acid group into the inner wall of the PEEK tube. As reaction conditions, the concentration of the sulfuric acid aqueous solution is preferably 50 wt% or more, the reaction temperature is preferably 15 ° C. to 80 ° C., and the reaction time is preferably 5 minutes to 7 days. However, when the sulfonation conditions become severe, the inner wall of the PEEK tube tends to be unevenly rough, and the number of theoretical plates tends to be difficult to be obtained. Therefore, the sulfuric acid aqueous solution of 50 wt% to 90 wt% is used for 5 minutes to 2 days at 15 ° C. to 60 ° C. The sulfonation conditions are preferred. After sulfonation, thoroughly wash with ion-exchanged water until neutrality, and then flow acetone or methanol into the PEEK tube and dry thoroughly with a nitrogen stream or a vacuum dryer.

(Vinyl group introduction process)
Next, at the time of forming the organic polymer monolith, a vinyl monomer having a glycidyl group is reacted with the sulfonic acid group so that the sulfonic acid group and the organic polymer monolith can be bridged (bonded). Thereby, a vinyl group is introduced into the inner wall of the PEEK tube via a sulfonyl group, particularly in this case via a sulfonyloxy group. Examples of vinyl monomers having a glycidyl group include acrylic acid having an epoxy group, derivatives of methacrylic acid, and derivatives of styrene having an epoxy group. For example, glycidyl acrylate, glycidyl methacrylate, glycidyl clonate, glycidyl itaconate Glycidyl fumarate, glycidyl malate, vinylbenzyl glycidyl ester and the like. As an introduction method, for example, a vinyl monomer having a glycidyl group is diluted with an organic solvent, injected into a resin tube into which a sulfonic acid group has been introduced, and both ends are sealed and reacted. As reaction conditions, the concentration of the vinyl monomer having a glycidyl group is preferably 0.05 mol / L to 10 mol / L, the reaction temperature is preferably 15 ° C. to 100 ° C., and the reaction time is 3 It is preferably ~ 48 hours. Examples of the organic solvent for dilution include diethyl ether, dimethyl sulfoxide, tetrahydrofuran, benzyl alcohol, chloroform, toluene, acetone, methylene chloride and the like. Methylene chloride, diethyl ether, and acetone are desirable from the viewpoint of easy compatibility with PEEK. After the reaction, the mixture is allowed to cool, washed with acetone or methanol, and then sufficiently dried with a nitrogen stream or a vacuum dryer.

  In FIG. 1, the schematic of the sulfonic acid group introduction | transduction process and vinyl group introduction | transduction process to PEEK is shown.

  Next, an example of a production process of an organic polymer monolith will be shown. In this step, a polymerizable monomer that is a raw material of the organic polymer monolith is polymerized in the resin tube into which the vinyl group has been introduced (organic polymer monolith polymerization step).

(Organic polymer monolith polymerization process)
In the present invention, the organic polymer monolith can be formed by a polymerization reaction of a polymerizable monomer having one or more polymerizable carbon-carbon double bonds in the molecule. For example, a copolymerization reaction of a polymerizable carbon-carbon double bond between a non-crosslinkable polymerizable monomer and a crosslinkable polymerizable monomer, or a polymerizable carbon-carbon bond of a crosslinkable polymerizable monomer For example, a double bond polymerization reaction.

  In the present invention, the non-crosslinkable polymerizable monomer used is a monomer having one polymerizable carbon-carbon double bond in one molecule. Specific examples include acrylic, methacrylic, styrene, and allyl non-crosslinkable polymerizable monomers, and acrylic and methacrylic non-crosslinkable polymerizable monomers are preferably used. Moreover, as a non-crosslinkable polymerizable monomer, the non-crosslinkable polymerizable monomer which has a hydroxyl group, an epoxy group, an alkyloxy group, an aryloxy group, an aralkyloxy group etc. other than an alkyl group is mentioned.

  Non-crosslinkable polymerizable monomers having an alkyl group include derivatives of acrylic acid and methacrylic acid having an alkyl group, such as methyl acrylate, ethyl acrylate, propyl acrylate, butyl acrylate, and acrylic acid. Pentyl, hexyl acrylate, heptyl acrylate, octyl acrylate, nonyl acrylate, decyl acrylate, undecyl acrylate, dodecyl acrylate, etc., methyl methacrylate, ethyl methacrylate, propyl methacrylate, butyl methacrylate, pentyl methacrylate Hexyl methacrylate, heptyl methacrylate, octyl methacrylate, nonyl methacrylate, decyl methacrylate, undecyl methacrylate, dodecyl methacrylate, lauryl methacrylate and the like.

  Examples of the non-crosslinkable polymerizable monomer having a hydroxyl group include acrylic acid and methacrylic acid derivatives having a hydroxyl group, such as hydroxyethyl acrylate, hydroxypropyl acrylate, hydroxybutyl acrylate, 2-hydroxy acrylate. -3-phenyloxypropyl, 2-hydroxyethyl acrylate, 4-hydroxybutyl acrylate, neopentyl glycol monoacrylate, etc., hydroxyethyl methacrylate, hydroxypropyl methacrylate, hydroxybutyl methacrylate, 2-hydroxy methacrylate Examples include -3-phenyloxypropyl, 2-hydroxyethyl methacrylate, 4-hydroxybutyl methacrylate, neopentyl glycol monomethacrylate, and the like.

  Examples of the derivative of the non-crosslinkable polymerizable monomer having an epoxy group include acrylic acid having an epoxy group, a derivative of methacrylic acid, and a derivative of styrene having an epoxy group. For example, glycidyl acrylate, glycidyl methacrylate Glycidyl clonate, glycidyl itaconate, glycidyl fumarate, glycidyl malate, vinylbenzyl glycidyl ester and the like.

  Non-crosslinkable polymerizable monomers having an alkyloxy group include acrylic acid and methacrylic acid derivatives having an alkyloxy group, such as methoxyethyl acrylate, ethoxyethyl acrylate, propoxyethyl acrylate, acrylic Butoxyethyl acid, methoxydiethylene glycol acrylate, ethoxydiethylene glycol acrylate, methoxyethylene glycol acrylate, butoxytriethylene glycol acrylate, methoxydipropylene glycol acrylate, methoxyethyl methacrylate, ethoxyethyl methacrylate, propoxyethyl methacrylate , Butoxyethyl methacrylate, methoxydiethylene glycol methacrylate, ethoxydiethylene glycol methacrylate, methoxyethyl methacrylate Glycol, methacrylic acid butoxy triethylene glycol, methacrylic acid methoxy dipropylene glycol.

  Examples of the non-crosslinkable polymerizable monomer having an aryloxy group or an aralkyloxy group include acrylic acid and methacrylic acid derivatives having an aryloxy group or an aralkyloxy group. For example, phenoxyethyl acrylate, acrylic Examples include acid phenoxydiethylene glycol, phenoxytetraethylene glycol acrylate, phenoxyethyl methacrylate, phenoxydiethylene glycol methacrylate, phenoxytetraethylene glycol methacrylate, and the like.

Furthermore, examples of the non-crosslinkable polymerizable monomer having other functional groups that can be used include benzyl acrylate, benzyl methacrylate, cyclohexyl acrylate, cyclohexyl methacrylate, tetrahydrofurfuryl acrylate, and tetrahydrofurfuryl methacrylate. Dicyclopentenyl acrylate, dicyclopentenyl methacrylate, dicyclopentenyloxyethyl acrylate, dicyclopentenyloxyethyl methacrylate, N-vinyl-2-pyrrolidone acrylate, N-vinyl-2-pyrrolidone methacrylate, etc. Can be mentioned.
The non-crosslinkable polymerizable monomer is not limited to these specific examples, and may be used by mixing several kinds thereof.

On the other hand, the crosslinkable polymerizable monomer used in the present invention may be any monomer as long as it has two or more polymerizable carbon-carbon double bonds in one molecule. Examples of the monomer having two polymerizable groups in one molecule include divinylbenzene, glycol and methacrylic acid or acrylic acid diesters such as ethylene glycol dimethacrylate, ethylene glycol diacrylate, propylene glycol dimethacrylate, Propylene glycol diacrylate, 1,3-butylene glycol dimethacrylate, 1,3-butylene glycol diacrylate, 1,4-butanediol dimethacrylate, 1,4-butanediol diacrylate, 1,5 -Pentanediol dimethacrylate, 1,5-pentanediol diacrylate, 1,6-hexanediol dimethacrylate, 1,6-hexanediol diacrylate, neopentylglycol dimethacrylate, neopentylglycol Diacrylate, diethylene glycol dimethacrylate, diethylene glycol diacrylate, triethylene glycol dimethacrylate, triethylene glycol diacrylate, polyethylene glycol dimethacrylate, polyethylene glycol diacrylate, tripropylene glycol dimethacrylate, tripropylene glycol diacrylate, trimethylolpropane di There are methacrylate, trimethylol propane diacrylate, tetramethylol methane dimethacrylate, tetramethylol methane diacrylate, polypropylene glycol dimethacrylate, polypropylene glycol diacrylate, etc. As monomers having 3 or more polymerizable groups in one molecule For example, trimethylolpropane tri Tacrylate, trimethylolpropane triacrylate, trimethylolethane trimethacrylate, trimethylolethane triacrylate, tetramethylolmethane trimethacrylate, tetramethylolmethane triacrylate, pentaerythritol trimethacrylate, pentaerythritol triacrylate, pentaerythritol tetramethacrylate, pentaerythritol tetra An acrylate etc. are mentioned.
The crosslinkable polymerizable monomer is not limited to these specific examples, and may be used by mixing several kinds thereof.

  The monolith used in the present invention is preferably obtained by copolymerizing the non-crosslinkable polymerizable monomer with the crosslinkable polymerizable monomer. Alternatively, the monolith used in the present invention can be obtained by polymerizing only a crosslinkable polymerizable monomer. When a non-crosslinkable polymerizable monomer and a crosslinkable polymerizable monomer are used in combination, the blending ratio of both is 50 to 50 of the above non-crosslinkable polymerizable monomer with respect to the total amount of polymerizable monomers. It is preferable that 90% by weight and the crosslinkable polymerizable monomer be 10 to 50% by weight.

  When the crosslinkable polymerizable monomer is less than 10% by weight, the resulting monolith has poor mechanical strength, poor durability in repeated use, and tends to make stable detection and analysis difficult. On the other hand, if the crosslinkable polymerizable monomer exceeds 50% by weight, the pore diameter tends to be difficult to adjust in some cases.

A polymerization initiator can be used for the polymerization for producing the monolith. The polymerization initiator used is preferably a peroxide radical polymerization initiator or an azo radical polymerization initiator, such as benzoyl peroxide, 2-ethylhexyl perbenzoate, acetyl peroxide, isobutyryl peroxide, or octanoyl peroxide. , Lauroyl peroxide, ditert-butyl peroxide, cumene hydroperoxide, methyl ethyl ketone peroxide, 4,4,6-trimethylcyclohexanone ditert-butylperoxyketal, cyclohexanone peroxide, methylcyclohexanone peroxide, acetylacetone peroxide, cyclohexanone di-tert-butyl Peroxy radicals such as peroxyketal, acetone di-tert-butyl peroxyketal, diisopropyl hydroperoxide, 2,2′-azobi Swisobutyronitrile, 2,2′-azobis (2,4-dimethylvaleronitrile), (1-phenylethyl) azodiphenylmethane, 2,2′-azobis (4-methoxy-2,4-dimethylvaleronitrile), dimethyl -2,2'-azobisisobutyrate, 2,2'-azobis (2-methylbutyronitrile), 1,1'-azobis (1-cyclohexanecarbonitrile), 2- (carbamoylazo) isobutyronitrile Azo radicals such as 2,2′-azobis (2,4,4-trimethylpentane), 2-phenylazo-2,4-dimethyl-4-methoxyvaleronitrile, 2,2′-azobis (2-methylpropane) A polymerization initiator is mentioned.
As a polymerization initiator, it is not limited to these specific examples, Furthermore, several of these can also be mixed and used.

  When a polymerization initiator is used, the polymerization initiator is preferably used in an amount of 0.05 to 10 parts by weight with respect to 100 parts by weight of the polymerizable monomer. When the amount used is less than 0.05 parts by weight, the polymerization time tends to be long, and unreacted monomers tend to remain in the organic polymer monolith. On the other hand, when the amount used exceeds 10 parts by weight, not only the polymerization initiator is wasted, but also heat generation control during polymerization is difficult and problems such as insufficient molecular chain length tend to occur.

  Usually, in order to maintain the porosity of the monolith, various solvents are further added during polymerization as a pore regulator. The solvent is preferably a solvent capable of dissolving the polymerizable monomer but not the resulting polymer. Specifically, toluene, xylene, ethylbenzene, diethylbenzene, heptanol, isoamyl alcohol, propanol, 1,4-butane. Known compounds such as diol, ethyl acetate, butyl acetate, dimethyl phthalate, diethyl phthalate, and other aliphatic or aromatic esters, ethylene glycol monoethyl ether acetate, hexane, octane, decane and the like can be used. These are properly used depending on the type of monomer used as the raw material of the organic polymer monolith to be obtained, and may be used alone or in combination of several types.

  The mixing ratio of these solvents is preferably 5 to 300% by weight, more preferably 20 to 200% by weight, and still more preferably 50 to 100% by weight based on the total amount of the polymerizable monomers from the viewpoint of porosity. The If the blending ratio is less than 5% by weight or exceeds 300% by weight, the desired porosity tends to be difficult to obtain.

  The monolith used in the present invention is obtained by polymerizing a composition containing the above polymerizable monomer and, if necessary, a polymerization initiator and a pore regulator in a PEEK tube having a sulfonic acid group introduced therein. be able to. General conditions are that the polymerization temperature is 50 ° C. to 100 ° C., and the polymerization time is 3 to 24 hours.

As described above, in the production method described above, the sulfonic acid group introduction step, the vinyl group introduction step, and the organic polymer monolith polymerization step were performed in this order, but as another method, after the sulfonic acid group introduction step, the organic polymer monolith polymerization step It is also possible to use the method of performing.
Specifically, first, a sulfonic acid group is introduced into the inner wall of the resin tube (sulfonic acid group introduction step). Thereafter, a polymerizable monomer that is a raw material of the organic polymer monolith is polymerized in the resin tube into which the sulfonic acid group has been introduced (organic polymer monolith polymerization step). In this case, the polymerizable monomer can be selected from the polymerizable monomers having a polymerizable carbon-carbon double bond in the molecule described above. A polymerizable monomer capable of reacting with the group is contained. The polymerizable monomer capable of reacting with the sulfonic acid group makes it possible to form a bridge (bond) between the sulfonic acid group and the organic polymer monolith. As the polymerizable monomer that can react with the sulfonic acid group, for example, the above-mentioned non-crosslinkable polymerizable monomer having an epoxy group can be used.

  In the present invention, the monolith column using the above-mentioned non-crosslinkable polymerizable monomer having an alkyl group exhibits capability as a hydrophobic high performance liquid chromatography (HPLC) column because the alkyl chain is hydrophobic.

  Moreover, the monolith column using the non-crosslinkable monomer which has free carboxylic acid groups, such as acrylic acid and methacrylic acid, shows capability as a cation exchange HPLC column, since a carboxylic acid group is acidic.

  In addition, monolithic columns using a non-crosslinkable monomer having a hydroxyl group and a glycidyl group can easily introduce amino groups, sulfonic acid groups, etc., and monolithic columns into which these groups are introduced are anion exchange, It can be used as a cation exchange HPLC column.

  EXAMPLES Hereinafter, although the column for liquid chromatography of the present invention and the production method thereof will be specifically described with reference to examples, the present invention is not limited to the examples.

Example 1
<Sulfonation of PEEK tube>
A PEEK tube (NPK-026 manufactured by Nirei Kogyo Co., Ltd.) having an inner diameter of 1 mm and a length of 10 cm is filled with 90% sulfuric acid aqueous solution, sealed at both ends, reacted at 40 ° C. for 10 minutes, and then 5 ml of ion-exchanged water, acetone 5 ml was poured into the tube for cleaning, and purged with nitrogen and dried.

  Table 1 shows the results of measuring the elemental composition ratio of the surface of the inner wall of the PEEK tube with an energy dispersive X-ray spectrometer (EDX) (Inca Energy 350, manufactured by Oxford Instruments Co., Ltd.). The content of 1.1% of S atoms (number of atoms) was confirmed, and it was found that sulfonic acid groups were introduced into the inner wall of the PEEK tube.

<Introduction of vinyl group>
Next, 1 mol / L glycidyl methacrylate / methylene chloride solution is filled into the above tube, both ends are sealed, reacted at 25 ° C. for 24 hours, then washed with 5 ml of acetone flowing in the tube and purged with nitrogen. And dried.

<Monolith polymerization>
Lauryl methacrylate 312.8 mg, ethylene glycol dimethacrylate 42 mg, 1-propanol 305.8 mg, 1,4-butanol 221.5 mg, and azobisisobutyronitrile 4 mg were mixed in a flask to Dissolved with a sonic cleaner. After sulfonation, the above-mentioned mixed monomer solution was injected into a PEEK tube into which a vinyl group was introduced with a syringe, stoppered so as not to leak both ends, and heated at 60 ° C. for 24 hours in a water bath.

<Evaluation>
The obtained column was cut into pieces and observed with SEM (SU-70, manufactured by Hitachi High-Technologies Corporation).
Further, one end of the obtained column was connected to a high performance liquid chromatography pump, the other end was opened, and water / acetonitrile 50:50 (volume ratio) was passed therethrough. A pressure of 15 MPa was applied to confirm whether the monolith did not escape from the column.
Furthermore, the measurement was performed using high performance liquid chromatography (L2000, manufactured by Hitachi High-Technologies Corporation) under the following conditions.
Eluent: Water / acetonitrile 50:50 (volume ratio)
Flow rate: 0.25 ml / min
Measurement wavelength: 214 nm (semi-micro cell)
Sample: Uracil (1 mM)
Methylbenzene, ethylbenzene, propylbenzene, butylbenzene, pentylbenzene (each 10 mM)
Injection volume: 1 μm

As a result of SEM observation, it was confirmed that the tube and the organic polymer monolith were in close contact (FIG. 3).
In addition, even when liquid was passed with one end of the obtained column open, the monolith did not escape from the tube, and it was confirmed that the pressure resistance was 15 MPa or more.
In the evaluation by HPLC, top peak separation of 6 components was achieved (FIG. 2). Moreover, the theoretical plate number of ethylbenzene became 4000 steps. The evaluation results are shown in Table 2.

(Example 2)
<Sulfonation of PEEK tube>
A PEEK tube (NPK-026 manufactured by Nirei Kogyo Co., Ltd.) having an inner diameter of 1 mm and a length of 10 cm is filled with 80% sulfuric acid aqueous solution, sealed at both ends, reacted at 25 ° C. for 24 hours, and then 5 ml of ion-exchanged water, acetone 5 ml was poured into the tube for cleaning, and purged with nitrogen and dried.
<Introduction of vinyl group>
Next, 1 mol / L glycidyl methacrylate / acetone solution is filled into the above tube, both ends are sealed, reacted at 25 ° C. for 24 hours, then washed with 5 ml of acetone flowing in the tube and purged with nitrogen. Dried.
<Monolith polymerization>
312.8 mg of lauryl methacrylate, 42 mg of ethylene glycol dimethacrylate, 305.8 mg of 1-propanol, 221.5 mg of 1,4-butanol, and 4 mg of azobisisobutyronitrile were mixed in a flask, and azobisisobutyronitrile was mixed. Dissolved with a sonic cleaner. After sulfonation, the above-mentioned mixed monomer solution was injected into a PEEK tube into which a vinyl group was introduced with a syringe, stoppered so as not to leak both ends, and heated at 60 ° C. for 24 hours in a water bath.

  Evaluation was carried out according to Example 1. As a result of SEM observation, it was confirmed that the tube and the organic polymer monolith were in close contact with each other. In addition, even when liquid was passed with one end of the obtained column open, the monolith did not escape from the tube, and it was confirmed that the pressure resistance was 15 MPa or more. According to the evaluation by HPLC, the top peak of 6 components could be separated. Moreover, the theoretical plate number of ethylbenzene became 4000 steps. The evaluation results are shown in Table 2.

(Example 3)
A PEEK tube (NPK-026 manufactured by Nirei Kogyo Co., Ltd.) having an inner diameter of 1 mm and a length of 100 cm was used as the PEEK tube. Otherwise, the column was prepared according to the method of Example 1, and then evaluated in the same manner as in Example 1.
As a result of SEM observation, it was confirmed that the tube and the organic polymer monolith were in close contact with each other. In addition, even when liquid was passed with one end of the obtained column open, the monolith did not escape from the tube, and it was confirmed that the pressure resistance was 15 MPa or more. According to the evaluation by HPLC, the top peak of 6 components could be separated. Moreover, the theoretical plate number of ethylbenzene became 7000. The evaluation results are shown in Table 2.

(Comparative Example 1)
Without performing the steps of sulfonation and vinyl group introduction, monolith polymerization was performed according to the method of Example 1 using a PEEK tube (NPK-026 manufactured by Nirei Kogyo Co., Ltd.) having an untreated inner diameter of 1 mm and a length of 10 cm. Prepared and evaluated.
As a result of SEM observation, it was confirmed that there was a gap due to resin shrinkage between the tube and the organic polymer monolith (FIG. 4).
Moreover, S atom was not detected in EDX, and when water / acetonitrile was allowed to flow through the column at a flow rate of 0.05 ml / min, the monolith was released from the tube at 2 MPa. The evaluation results are shown in Tables 1 and 2.

(Comparative Example 2)
A PEEK tube (NPK-026 manufactured by Nirei Kogyo Co., Ltd.) having an inner diameter of 1 mm and a length of 10 cm is filled with a 90% sulfuric acid aqueous solution, sealed at both ends, reacted at 25 ° C. for 2 hours, and then 5 ml of ion-exchanged water and acetone. 5 ml was poured into the tube for cleaning, and purged with nitrogen and dried.
Next, a monolithic column was prepared according to Example 1 and evaluated without carrying out the vinyl group introduction reaction.
As a result of SEM observation, it was confirmed that there was a gap due to resin shrinkage between the tube and the organic polymer monolith. Further, when water / acetonitrile was allowed to flow at a flow rate of 0.05 ml / min, the monolith came out of the tube at 2 MPa. The evaluation results are shown in Table 2.

  In the liquid chromatography column of the present invention, the volume shrinkage of the organic polymer monolith hardly occurs when the organic polymer monolith is produced in the column. Therefore, the organic polymer monolith does not easily escape from the column during use. . Moreover, since the column and the organic polymer monolith were combined through the sulfonyl group using the tube formed with resin containing PEEK in the column, it was excellent in solvent resistance, acid resistance, and alkali resistance. The column for liquid chromatography of the present invention is a column suitable for high performance liquid chromatography (HPLC), and a high number of theoretical plates was obtained by using the column for liquid chromatography of the present invention.

Claims (5)

A resin tube, and an organic polymer monolith formed in the resin tube;
The resin comprises polyetheretherketone;
The column for liquid chromatography in which the organic polymer monolith is bonded to the resin tube via a sulfonyl group.   The column for liquid chromatography according to claim 1, wherein the organic polymer monolith is bonded to the resin tube via a sulfonyloxy group.   The column for liquid chromatography according to claim 1 or 2, wherein the organic polymer monolith is a monolith obtained by polymerizing a polymerizable monomer having a polymerizable carbon-carbon double bond in the molecule.   The column for liquid chromatography according to claim 1, wherein the organic polymer monolith is a (meth) acrylic resin monolith. Introducing a sulfonic acid group into the inner wall of the resin tube,
By reacting the introduced sulfonic acid group and a vinyl monomer having a glycidyl group, a vinyl group is introduced into the inner wall of the resin tube,
A polymerized monomer having a polymerizable carbon-carbon double bond in a molecule is polymerized in the resin tube into which a vinyl group has been introduced to form an organic polymer monolith in the resin tube. 5. A method for producing a liquid chromatography column according to 3 or 4.