JPH02196810A - Acrylic crosslinked polymer particle and production thereof - Google Patents

Abstract

PURPOSE:To obtain the subject particles for carrier of chromatography, etc., having narrow particle size distribution by dispersing acrylic reactive monomer, etc., into a solvent dissolving monomer and not dissolving polymer, polymerizing, subjecting crosslinkable monomer, etc., to seed polymerization and introducing functional group. CONSTITUTION:Acrylic reactive monomer, or said monomer and crosslinkable monomer are dispersed in a solvent dissolving raw material monomer and not dissolving generated polymer and polymerized to obtain seed polymer, then crosslinkable monomer in an amount of 1-20wt. times of said seed polymer, or said crosslinkable monomer and acrylic reactive monomer are subjected to seed polymerization to obtain a polymer containing 60-95wt.% crosslinkable monomer unit and 5-40wt.% acrylic reactive monomer unit, thus functional group in an amount of 40-100% of the reactive monomer unit is introduced into resultant system to afford the aimed polymer particle having <=10% standard deviation value and 1-30mu particle diameter range.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to acrylic crosslinked polymer particles and a method for producing the same.

The present invention relates to acrylic crosslinked polymer particles with high yield and a method for producing the same.

[Conventional technology]

Conventionally, polymers derived from natural products, synthetic polymer copolymers, silica gel, etc. have been used as carriers for chromatography.

[Problem to be solved by the invention]

All of these carriers have large particle diameters, high degree of swelling, and low mechanical strength, and therefore have the following drawbacks when used as carriers for liquid chromatography.

b) As the flow rate increases, the differential pressure and pressure increase increase, making it difficult to pass liquid at high speed.

Mouth) The number of theoretical plates is low.

7 It is not possible to create long columns.

2) Silica gel dissolves under weakly alkaline conditions.

e) When the degree of crosslinking is increased, the exchange capacity decreases.

An object of the present invention is to provide an acrylic crosslinked polymer useful as a carrier for liquid chromatography, which has improved the above-mentioned drawbacks.

[Means to solve the problem]

That is, in the present invention, (1) the weight fraction of crosslinkable monomer units is 60-9! r%, the weight fraction of the acrylic reactive monomer unit is 5-yo%, the functional group is introduced by θ to ioo% with respect to the reactive monomer unit, and the standard deviation value of the particle size distribution is 70% or less, and 7 to 30
The gist of the present invention is to provide crosslinked acrylic polymer particles having a particle size range of 4 m, and (2) a method for producing crosslinked acrylic polymer particles through the following steps.

First step: An acrylic seed polymer is produced by dispersing and polymerizing an acrylic reactive monomer or a crosslinking monomer and an acrylic reactive monomer in a solvent that dissolves the raw material monomer but does not dissolve the produced polymer.

Seed polymerization of the seed polymer produced in the seventh step and a crosslinkable monomer in an amount of 7 to 20 times the weight of the seed polymer, or a crosslinkable monomer and an acrylic reactive monomer, The weight fraction of crosslinkable monomer units is 6
An acrylic crosslinked raw material polymer having a weight fraction of 1-II 0% of acrylic reactive monomer units is produced.

Third step: Acrylic crosslinked polymer particles are manufactured by introducing a functional group into the acrylic crosslinked raw material polymer manufactured in the third step.

Examples of the acrylic reactive monomer of the present invention include ethylenically unsaturated carboxylic acid esters. Preferably glycidyl acrylate, glycidyl methacrylate, co-hydroxyethyl methacrylate, -1(
Examples include N,N'-dimethylamino) methacrylate and glycerin monomethacrylate.

Examples of crosslinkable monomers include polyvinyl esters of polyhydric alcohols.

Preferable examples include ethylene glycol diacrylate, ethylene glycol dimethacrylate, glycerin diacrylate, glycerin dimethacrylate, glycerin triacrylate, glycerin trimethacrylate, trimethylolpropane trimethacrylate, and polyethylene glycol dimethacrylate.

Next, a method for producing the acrylic crosslinked polymer particles of the present invention will be explained.

The acrylic crosslinked polymer particles are produced by producing an acrylic seed polymer in the first step, and in the second step,
The seed polymer produced in the first step is further impregnated with a crosslinkable monomer or a crosslinkable upper layer and an acrylic reactive monomer, and the seed polymer is enriched by polymerization. .

The degree of crosslinking of the acrylic seed polymer produced in the first step is 0% to 2.0%, preferably 0.0%. /%~/
, j%. Here, the degree of crosslinking refers to the weight fraction of crosslinkable monomers in all monomers.

However, when using a polymerizable monomer such as glycidyl acrylate or glycidyl methacrylate as the acrylic reactive monomer, a part of the monomer acts as a crosslinking monomer, so that the degree of crosslinking is less than the preferred range described above. Alternatively, a preferred seed polymer can be obtained without using a crosslinking monomer.

In the first step, in order to obtain polymer particles having the desired particle size, it is necessary to appropriately select the solubility parameter expressed by (ΔE/■) when the heat of vaporization of the medium used is 681 molar volume V. be. That is, the larger the difference between the solubility parameter of the produced polymer particles and the solubility parameter of the medium used, the smaller the particle size of the produced polymer particles tends to be. The medium used in the present invention has a solubility or higher/b, z (c
al/aI) - those in the following ranges are used. Specifically, methanol, ethanol, coprobanol,
Examples include alcohols such as co-methyl-copropanol, ethers/alcohols such as these alcohols-methoxyethanol, ethers such as tetrahydrofuran and dioxane, and mixtures of these various solvents with water.

Furthermore, during polymerization of the seed polymer (dispersion polymerization), the particle size of the polymer particles can be controlled by using a diluent in the medium as necessary. The amount of the diluent used can be changed depending on the type and amount of the polymerizable monomer, but it is usually preferably used at 30% (by weight) or less, and the solubility parameter of the diluent is 1. θ(ca
l/cit)X or more and q,o (cal/ad)- or less. Specific examples of diluents include cyclohexane and xylene.

During polymerization of the seed polymer, it is preferable to use a dispersion stabilizer to prevent agglomeration, deformation, and fusion of the resulting polymer particles and to increase their dispersion stability. As the dispersion stabilizer, synthetic polymer compounds such as various homopolymer copolymers, graft polymers, and block polymers, as well as natural polymer compounds and their derivatives, etc. can be used.

Specific examples include polyvinylpyrrolidone, polyvinyl methyl ether, polyethyleneimine, polyacrylic acid, vinyl alcohol-vinyl acetate copolymer, ethyl cellulose, and hydroxypropyl cellulose.

Furthermore, during the polymerization of the seed polymer, a co-stabilizer can be used in addition to the above-mentioned dispersion stabilizer in order to make the polymerization proceed more stably and improve dispersion stability. As such co-stabilizers, anionic surfactants, nonionic surfactants, secondary ammonium salts, long-chain alcohols, etc. are used. Specific examples include sodium di(λ-ethylhexyl)sulfosuccinate, nonylphenoxypolyethoxyethanol, methyltricafrylylammonium chloride, and cetyl alcohol.

Further, as polymerization initiators that can be preferably used in the polymerization of the seed polymer, co, -'-7zobisimptilonitrile, p, u'-azobis-(lI-cyanopentanoic acid), 2,2'- Examples include azo polymerization initiators such as azobis-(-monomethylbutyronitrile) and 2,2'-azobis-(co,4!-dimethylvaleronitrile), and peroxides such as benzoyl peroxide.

Further, the polymerization temperature during seed polymer polymerization in the present invention is lθ°C to 0°C, preferably 60°C to tro°C.
can be mentioned.

Next, a seed polymerization method is used for the second step, that is, for enlarging the seed polymer produced in the first step. .

These seed polymers can be used in the form of a dispersion such as an emulsion or suspension.

The particle size of the seed polymer can be arbitrarily selected within the range of 1/3 or more of the particle size of the target polymer, and the polymer particles obtained in the present invention can be used as a seed polymer in the next step. , even larger polymer particles can also be produced.

Examples of the polymerization initiator that can be used in the second step of the present invention include λ, 2'-azobisisobutyronitrile,
'l+'l' 7zobis(lI-cyanopentanoic acid)
, x, 2'-azobis(2-methylbutyronitrile),
x, azo polymerization initiator such as 2'-azobis(2,4(-dimethylvaleronitrile), benzoyl peroxide, lauroyl peroxide, octanoyl peroxide 1.),
J',! ; - Peroxides such as trimethylhexanoyl peroxide and the like can be mentioned.

These polymerization initiators can be used alone or in combination.

In the second step, the seed polymer produced in the first step is enlarged, that is, seed polymerized, and the weight fraction of the crosslinkable monomer in the monomers charged in this seed polymerization is AO% to lθθ%.

Furthermore, porous polymer particles can be obtained by adding a suitable diluent to the monomer phase during seed polymerization. By making it porous, the amount of functional groups that can be introduced increases and the performance as a preparative chromatography carrier is improved.

As the diluent, it is preferable to use an organic solvent that has a low swelling property for the produced polymer particles. Further, in preparing the dispersion liquid, it is preferable to use a dispersion stabilizer in order to improve dispersibility. Known dispersion stabilizers can be used as such dispersion stabilizers, including anionic surfactants such as sodium dodecylbenzenesulfonate, sodium lauryl sulfate, sodium dialkylsulfosuccinate, and polyoxyethylene nonylphenyl. Nonionic surfactants such as ether, polyethylene glycol monostearate, and sorbitan monostearate, and synthetic polymer compounds such as polyvinylpyrrolidone, polyethyleneimine, and vinyl alcohol-vinyl acetate copolymers can be used in combination.

In the present invention, during seed polymerization, a dispersion stabilizer is used to prevent agglomeration, deformation, and fusion of the seed polymer that has become swollen by absorbing the polymerization initiator and polymerizable monomer, and to increase its dispersion stability. It is necessary to use Known anionic and nonionic surfactants can be used as such dispersion stabilizers, and these can also be used in combination with synthetic polymer compounds such as polyvinylpyrrolidone, polyethyleneimine, vinyl alcohol-vinyl acetate copolymer, etc. It is.

The polymerization temperature in the seed polymerization of the present invention is usually DaO
C to 90C, preferably 50C to goC.

Next, a functional group is introduced in a third step.

The acrylic crosslinked polymer particles of the present invention into which functional groups have been introduced in the third step can be used as a chromatography carrier for both analysis and preparative separation.

When the crosslinked polymer particles of the present invention are used as a chromatography carrier, applicable separation forms include ion exchange chromatography, hydrophobic chromatography,
Examples include reverse phase chromatography.

In addition, when the crosslinked polymer particles of the present invention are used as a carrier for Chromadog 2F, the type of functional group introduced into the particles depends on the above-mentioned chromatography format. (a) Ion exchange chromatography: sulfopropyl group, carboxy Methyl group, dimethylaminomethyl group,
(b) Hydrophobic chromatography: phenyl group-butyl group, polyoxyethylene group, etc. (c) Reversed phase chromatography: octyl group, octadecyl group, etc. Chromatography of the acrylic crosslinked polymer particles of the present invention When used as a carrier, a functional group is introduced into the acrylic crosslinked raw material polymer produced in the λth step. The method differs depending on each chromatography type, but for example, when the reactive monomer unit has an epoxy group, , a method in which an epoxy group is ring-opened and modified with water, glycerin, or ethylene glycol, and each of the above-mentioned functional groups is bonded to the generated hydroxyl group, or a method in which a functional group is directly bonded to the epoxy group.

In the acrylic crosslinked polymer particles of the present invention produced by the above steps, the weight fraction of crosslinkable monomer units and the weight fraction of reactive monomer units can be analyzed by pyrolysis gas chromatography. The rate is tr.

% or more, it is recognized that it almost matches the charging ratio of raw material monomers.

In addition, methods for measuring the rate of functional group introduction include ion exchange capacity for ion exchange groups, nitrogen content by elemental analysis for functional groups with nitrogen atoms, and C1C13-N for functional groups with benzene rings. Alternatively, it can be measured by FT-IR, but other functional groups can also be measured by pyrolysis gas chromatography, mass spectrometry, FT-IR, elemental analysis, etc.

[For production]

In the preferred production method of the acrylic crosslinked polymer particles of the present invention, since the particle size of the seed polymer obtained in the first step is large, the degree of enlargement in the seed polymerization in the λth step is 2θ times or less in volume. can be lowered,
The properties of the seed polymer can be reflected in the enlarged particles. That is, since the particle size of the seed polymer 0 is uniform,
The particle size of the resulting crosslinked polymer particles is also uniform. Also,
In order to absorb and polymerize a polymerizable monomer containing a low polarity crosslinking polymer into a seed polymer containing many polar reactive monomer units, the polar reactive monomer units tend to gather on the surface layer of the particles, resulting in Despite K having a high degree of crosslinking, polymer particles with a high functional group introduction rate can be obtained. Such features of the crosslinked polymer particles of the present invention are unprecedented.

The polymer particle fraction produced by the method according to the invention is S%
An acrylic cross-linked material having a particle size range of 7 to 303 m, and a standard deviation of particle size distribution of 19% or less. They are polymer particles.

Furthermore, when the particles are used as a chromatography carrier, they provide extremely excellent separation properties.

〔Example〕

EXAMPLES The present invention will be explained below with reference to Examples, but the present invention is not limited to these Examples.

Example 1 (1) Production of acrylic crosslinked polymer particles -jO (manufactured by Tokyo Kasei Co., Ltd., average molecular weight 0.000)/, tri, glycidyl methacrylate/J, jI, ethylene glycol dimethacrylate 0. 0II! and co, λ'-azobisin butyronitrile 0. /, 79 to ethanol 73.01 and cyclohexane/2. ! ; , 9 was dissolved in a mixed solvent, and polymerization was carried out at 70° C. for 3 hours under a nitrogen gas stream. The polymerization conversion rate in this polymerization was 9 g%. Furthermore, when the polymer particles obtained by this polymerization were observed with a scanning electron microscope, it was confirmed that they were truly spherical particles with a particle size of 1.6 μm and 1 standard deviation i,o% and highly monodisperse.

Using this as a seed polymer, the following operations were performed.

First, 2.2'-azobisweetsbutyronitrile 0.3A
Ii, ethylene glycol dimethacrylate 3b,
A I,/, 2-dichloroethane ti, y, t II
.

Sodium dodecyl sulfate 0.6 & , 51 and water 21
0 g was finely dispersed so that the particle size of oil droplets was 0.5 μm or less. This emulsion was mixed into an aqueous dispersion consisting of a monodisperse glycidyl methacrylate/ethylene glycol dimethacrylate copolymer s, og of particle size/j μm obtained in the previous polymerization, sodium dodecyl sulfate 0.0/p, and water/2'lli. It was added dropwise to the solution at room temperature over one hour, and the oil droplets were absorbed into the seed polymer while stirring slowly at the same temperature for 72 hours.

Next, add 30% polyvinyl alcohol aqueous solution to this. !
9 was added, and polymerization was carried out at 70° C. for t hours under a nitrogen gas flow. Final polymerization conversion rate? 3%, and the weight fraction of crosslinkable monomer units in the crosslinked polymer particles considering the polymerization conversion rate is 37%, and the weight fraction of reactive monomer units derived from glycidyl methacrylate is 13%, which is due to thermal decomposition. It showed good agreement with the analysis results by gas chromatography, and when measured by BET method,
It was a porous particle with a specific surface area t o, so m / ti.

When the polymer particles obtained by this polymerization were observed using a scanning electron microscope, they had a particle size of 3.2 μm and were highly monodisperse with a standard deviation of %.

(2) Ring opening with water (10 g of the polymer obtained in step 11 was diluted with 10% I (2S O
4 in aqueous solution 10711 at a temperature of go °C.
Hydrolysis was carried out by heating for a period of time, and the glycidyl groups in the polymer were ring-opened.

After the reaction was completed, the polymer was filtered off and thoroughly washed with demineralized water until the filtrate became neutral, thereby quantitatively obtaining a crosslinked acrylic resin having hydroxyl groups.

(3) Introduction of diethylaminoethyl group 1oti of ring-opening modified crosslinked acrylic resin obtained in 12)
The mixture was poured into an aqueous N-NaOH solution λil, and stirred for 6θ minutes at room temperature under ultrasonic dispersion. Then add 3 to this
i% β-diethylaminoethyl chloride hydrochloride aqueous solution 2
1:,69 was added and kept at goC for S hours.

After the reaction is completed, the filtrate is washed with demineralized water lθθ9 and then with iN-hydrochloric acid aqueous solution 209, and then thoroughly washed with demineralized water until the filtrate becomes neutral, and the amount of the acrylic anion exchange resin having diethylaminoethyl groups is determined. I got it.

The exchange capacity of the anion exchange resin manufactured with the above formulation is 0.
It was 5 meq/I-resin.

The functional group introduction rate with respect to the reactive monomer units in the polymer calculated from this value was 69%.

The exchange capacity of the obtained anion exchange resin was measured as follows:
This was done by the following method.

Wet product of anion exchange resin about -009/N-NaOH
The mixture was poured into an aqueous solution 20- and shaken at room temperature for 30 minutes. After filtering this, it was thoroughly washed with a pH 10 NaOH aqueous solution.

Next, the obtained anion exchange resin was dispersed in a 1% NaC6 aqueous solution, and an automatic titration device (GT-06 manufactured by Mitsubishi Kasei Corporation) was used under stirring to obtain o, oz N-HC.
1 aqueous solution at a dropping rate of 20 μl/sec, and a titration curve (Figure 1) was created. After the titration was completed, the anion exchange resin was filtered out, thoroughly washed with water, dried at goC for 3 hours, and its mass was weighed.

In Fig. 1, the horizontal axis is the dripping amount, and the vertical axis is the titration curve representing pH.
The exchange capacity was expressed by the following formula, where f was the factor of the sN-HCI aqueous solution and Mg was the mass of the dry anion exchange resin.

The obtained acrylic anion exchange resin having a diethylaminoethyl group was separated by ion exchange chromatography using a gradient elution method for a mixed protein containing myoglobin, neobarpmin, and trypsin inhibitor.

The chromatogram is shown in FIG. 2, and the obtained chromatogram shows a good degree of separation.

The device used for ion exchange chromatography using a gradient elution method was an LC-6A system manufactured by Shimadzu Corporation. Moreover, the measurement conditions are as shown below.

Measurement conditions column: 7. tmm IDX 75mm flow rate: /
, Od/min Elution: A − + 820 min Linear gradient A: /4 was added to the plate 01-trishydroxymethylaminomethane/hydrochloric acid buffer (pHg, 0) B: A + 0. ! r mol Sodium chloride ratio: UV 2 t Onm Temperature: 23°C In addition, the pressure during liquid passage under these measurement conditions is b ” ”
9/ci! The particle size was much lower than that of similar particle sizes produced by conventional suspension polymerization methods.

Example λ (1) Preparation of acrylic crosslinked polymer Polyvinylpyrrolidone, 130 t, trtt.

Glycidyl methacrylate 12. ! 11. Ethylene glycol, cold dimethacrylate) o, o o s # and 2.2'-7 zopis isoptylonitrile θ, 131 with ethanol? J, 011, cyclohexane lλ, 1
It was dissolved in a mixed solvent of Ii and polymerized at 70° C. for 3 hours under a nitrogen gas stream. The polymerization conversion group in this polymerization is 9
It was 1%. When the polymer particles obtained by polymerization with Sarak were observed with a scanning electron microscope, the particle size /,
It was confirmed that the particles were truly spherical particles with a diameter of 41 μm and a standard deviation of 1.9%, with extremely high monodispersity. Using this as a seed polymer, the following operations were performed.

First of all, J! '-Azobisinbutyronitrih0.36
1. Ethylene glycol dimethacrylic V-toj J, 9
g, ri) serine monomethacrylate 3.79, l-
Monodichloroethanda J, l: I, sodium dodecyl sulfate 0. bl, g and water-501 when the particle size of the oil droplet is 0
It was finely dispersed so that the distance was 0.054 m or less. This dispersion was converted into a monodispersed glycidyl methacrylate/ethylene glycol dimethacrylate copolymer s, op, sodium dodecyl sulfate θ, 0, with a particle size of the original polymerization/, lI μm.
/ Aqueous dispersion 11C consisting of 9 and water lλlIy
Dropwise over 2 hours at room temperature, then 1 hour at that temperature.
The oil droplets were absorbed into the seed polymer with slow stirring for 2 hours.

Then this KJ% polyvinyl alcohol and aqueous solution, 7
0.4 t, 9 was added, and polymerization was carried out at 70° C. for S hours under a nitrogen gas stream to obtain particles with a final polymerization conversion rate of 90%.

The weight fraction of crosslinkable monomer units in the crosslinked polymer particles in consideration of polymerization conversion ratio is 36%, and the weight fraction of reactive monomer units is 17%.

When the polymer particles obtained by this polymerization were observed using a scanning electron microscope, they had a particle size of 3.2 μm and were highly monodisperse with a standard deviation of 0%.

(2) Ring opening with water The polymer ioy obtained in step (1) is kept in a 10% H, So4 aqueous solution 1ooy at 50°C for 3 hours to perform hydrolysis. Ring opening was performed.

After the reaction was completed, the polymer was filtered off and thoroughly washed with demineralized water until the filtrate became neutral, thereby quantitatively obtaining a crosslinked acrylic resin having hydroxyl groups.

(Introduction of 314′ amino group (2) K) The ring-opening modified crosslinked acrylic resin 109 obtained from K
g, 09, and stirred for 30 minutes under ultrasonic dispersion at room temperature.

Then, to this, a water-containing p-)luenesulfonic acid l aqueous product-
05I was added and the reaction was carried out at 70°C for 2 hours. The reaction was filtered and washed with methanol.

The reactant 109 obtained above was mixed with trimethylamine/m
The mixture was placed in an aqueous solution of ol/- and reacted at 50°C for 3 hours. The reaction product was filtered, then washed with demineralized water, and air-dried at room temperature for 1 hour to quantitatively obtain an acrylic anion exchange resin.

The exchange capacity of the anion exchange resin manufactured with the above formulation is o,
bb meq/g-resin, and the functional group introduction rate with respect to the reactive monomer units in the polymer calculated from this value was 67%.

Furthermore, when the anion exchange resin produced according to the above formulation was used as a protein separation agent, extremely good separation properties were obtained.

Example 3 Tl) Preparation of acrylic crosslinked polymer -30togg to polyvinylpyrrolidone.

Glycidyl methacrylate 3θ11 and 2.2'-azobisisobutyronitrile o, 31 with ethanol &7.
9 # and polymerization was carried out at 70° C. for S hours under a nitrogen gas stream. The polymerization conversion rate in this polymerization was 9 g%. Furthermore, when the polymer particles obtained by this polymerization were observed with a scanning electron microscope, it was confirmed that they were truly spherical particles with a particle size of 2.58 ml and a standard deviation of 2.5%, with extremely high monodispersity. Using this as a seed polymer, the following operations were performed.

Mass, co, 2'-azobisisobutyronitrile 0.36
g, ethylene glycol dimethacrylate j A, A
I,/,2-dichloroethane'I 3.211 Sodium dodecyl sulfate 0.4 A I9 and water 2g09 were finely dispersed so that the particle size of oil droplets was O.S μm or less. This emulsion was mixed with the particle size obtained in the previous polymerization, 5 μm monodispersed polyglycidyl methacrylate s, o y , and sodium dodecyl sulfate 0.0/1. The oil droplets were added dropwise to an aqueous dispersion of SOI and water at room temperature over a period of 4 hours, and then slowly stirred at the same temperature for 3 hours to allow the oil droplets to be absorbed into the seed polymer.

Next, 3% polyvinyl alcohol aqueous solution j O,
! ; 11 was added, and polymerization was carried out at 70° C. for S hours under a nitrogen gas stream. The polymerization conversion rate was 95%, and when the polymer particles obtained by this polymerization were observed using a scanning electron microscope, they were found to be extremely monodisperse with a particle size of S μm and a standard deviation of S,0%. The polymerization fraction of crosslinkable monomer units in the crosslinked polymer considering the polymerization conversion rate is 37%,
Furthermore, the weight fraction of reactive monomer units derived from glycidyl methacrylate was 73%, which showed good agreement with the analysis results by pyrolysis gas chromatography.

(2) Ring-opening with water (1) Polymer/Qjl obtained in 1O%l-12S
o.

Hydrolysis was carried out by heating in an aqueous solution TOGG at a temperature of go°C for 1 S hour, and the glycidyl groups in the polymer were ring-opened.

After the reaction was completed, the polymer was filtered off and thoroughly washed with demineralized water until the filtrate became neutral, thereby quantitatively obtaining a crosslinked acrylic resin having hydroxyl groups.

(3) Introduction of diethylaminoethyl group (2) The ring-opening modified crosslinked acrylic resin 109 obtained in step (2) was
-N a OH aqueous solution 2/g and stirred for 60 minutes under ultrasonic dispersion at room temperature. Then add this &
/% β-diethylethyne ethyl chloride hydrochloride aqueous solution J
t,/,i was added and held at 50°C for S time.

After the reaction, demineralized water / 00 g, followed by /N-HC
After washing with an aqueous solution θI, the filtrate was thoroughly washed with demineralized water until it became neutral, thereby quantitatively obtaining an acrylic anion exchange resin having a diethylaminoethyl group. The exchange capacity of the anion exchange resin manufactured with the above formulation is o, q 6
It was meq/'l-reSin. The functional group introduction rate with respect to the reactive monomer units in the polymer calculated from this value was 51%.

Example q (1) Production of acrylic crosslinked polymer -30/, 11/ for polyvinylpyrrolidone.

glycidyl methacrylate) J 011 and 2.2
'-Azobisinbutyronitrile 0.31 was dissolved in 7.9 F of ethanol A and heated at 70°C under a nitrogen gas stream.
Time polymerization was performed. The polymerization conversion rate in this polymerization was 9g
%Met. Furthermore, the polymer particles obtained by this polymerization were observed with a scanning electron microscope and had a particle size of 2. ! ;l
1m.

It was confirmed that the particles were truly spherical particles with extremely high monodispersity with a standard deviation of 1.2%. Using this as a seed polymer, the following operations were performed.

First, 1.1.2'-azobisinbutyronitrile 0.3
&&, glycidyl methacrylate)//I, ethylene glycol dimethacrylate 2 &, A, 9%/
, 2-dichloroethane y, y, g 1, sodium dodecyl sulfate 0.669 and water 210I until the particle size of the oil droplets is O
, Sμm or less. This emulsion was mixed with the monodisperse polyglycidyl methacrylate obtained in the previous polymerization with particle diameters λ and S μm. ;,0.9, sodium dodecyl sulfate 0.0/9, and water sop dropwise at room temperature over a period of - hours, and then slowly stirred at the same temperature for 3 hours to form oil droplets. It was absorbed into Z seed polymer. Next, 3% polyvinyl alcohol aqueous solution 3θ, S9 was added to this, and the mixture was heated at 70°C under a nitrogen gas stream.
Polymerization was carried out for 8 hours. The polymerization conversion rate is t%, the polymerization fraction of crosslinkable monomer units in the crosslinked polymer considering the polymerization conversion rate is 60%, and the weight fraction of reactive monomer units derived from glycidyl methacrylate is 90%, This showed good agreement with the analysis results by pyrolysis gas chromatography. When the polymer particles obtained by this polymerization were observed using a scanning electron microscope, the particle size was S.
It had extremely high monodispersity with a standard deviation of 1 μm and 0%.

(2) Introduction of diethylamino group Crosslinked acrylic polymer i obtained in (i).

I was dispersed in 1 Iott of methanol, 29 was added to diethylamine, and the mixture was reacted at 60° C. for 6 hours.

After the reaction was completed, the mixture was thoroughly washed with demineralized water, and then reacted again in 5 otttl of 10X sulfuric acid at 30° C. for 3 hours. After the reaction was completed, the resin was washed with demineralized water, regenerated with an aqueous N-water thorium solution, and then sufficiently pressure-washed with demineralized water to quantitatively obtain an anion exchange resin having a diethylamino group. Also, the exchange capacity of the anion exchange resin produced according to the above formulation is /,! fmeq/f-res in,
The functional group introduction rate with respect to the reactive monomer units in the polymer calculated from this value was 56%. Furthermore, excellent separation properties were obtained in the separation of mixed proteins using this resin.

Example 5 (1) Introduction of carboxymethyl group 10 f of ring-opening modified acrylic crosslinked polymer particles obtained in the same manner as in Example 3-+2) were placed in a NaOH aqueous solution SOt in Co. II, and subjected to ultrasonic dispersion at room temperature. Stirring was then performed for 60 minutes.

Next, a monochloroacetic acid/62 coprobanol solution QOrnl was stirred at SO°C, and after completion of the dropwise addition, j was reacted at 90°C for an hour. After the reaction is complete, demineralize water! ;o
0511/N-HCl! After washing with Oml, the filtrate was washed with demineralized water until it became neutral to quantitatively obtain an acrylic cation exchange resin having a carboxymethyl group.

The exchange capacity of the cation exchange resin produced with the above formulation is 0.
．． 1i! ; meq/f-resin. The functional group introduction rate with respect to the reactive monomer units in the polymer calculated from this value was 50%.

Example 6 (1) Example of introducing sulfopropyl group, ? - Add 3.9f of 25% sodium methylate methanol solution to tetrahydrofuran dispersion 2S2 of ring-opening modified acrylic crosslinked polymer particles iov obtained in the same manner as (2), and stir for 30 minutes at room temperature under ultrasonic dispersion. I did it. Next, add 25 tons of propane sultone to this.
Add and heat to 90℃! The reaction was carried out for r hours.

After the reaction, wash with 500 y of demineralized water and 0.2 M-Na
, Co, and aqueous solution 301, the filtrate was washed with demineralized water until it became neutral, and an acrylic cation exchange resin having a sulfopropyl group was quantitatively obtained. The exchange capacity of the cation exchange resin produced with the above formulation is 0. lIb me
It was q/f-resin. The functional group introduction rate for the reactive monomer units in the polymer calculated from this value is s
It was i%.

Example 7 (1) Ring opening with ethylene glycol Example 3-(
The acrylic crosslinked polymer particles 102 obtained in step 1) were dispersed in 2 g of ethylene glycol and 12 g of concentrated sulfuric acid.
was added, and the reaction was carried out at 70°C for 7 hours to add ethylene glycol to the glycidyl groups in the polymer. After the reaction was completed, the polymer was filtered and thoroughly washed with demineralized water until the filtrate became neutral.A. Acrylic crosslinked polymer particles ring-opened and modified with ethylene glycol were quantitatively obtained. .

(2) Introduction of diethylaminoethyl groups Modified acrylic crosslinked polymer particles 102 in Example 3- (3)
A diethylamine ethyl group was introduced in the same manner as above to quantitatively obtain an acrylic anion exchange resin having an exchange capacity of o and q rmeq/7-resin. The functional group introduction rate with respect to the reactive monomer units in the polymer calculated from this value was SO%.

Furthermore, when the anion exchange resin produced according to the above formulation was used as a protein separation agent, extremely good separation properties were obtained.

Example (1) Ring opening with glycerin Acrylic crosslinked polymer particles obtained in Example 3-(1) is dispersed in glycerin, 7/, j f,
Sulfuric acid l? was added and reacted at 70°C for 7 hours to add to the glycidyl groups in the polymer.

After the reaction was completed, the polymer was filtered off and thoroughly washed with demineralized water until the filtrate became neutral, thereby quantitatively obtaining acrylic crosslinked polymer particles ring-opened and modified with glycerin.

(2) Introduction of diethylamine ethyl group The acrylic crosslinked polymer particles ioy ring-opened and modified with glycerin obtained in (1) were introduced with diethylaminoether in the same manner as in Example J-(3) to increase the exchange capacity. o, e b m
An anion exchange resin of eq/P-resin was quantitatively obtained. The functional group introduction rate with respect to the reactive monomer units in the polymer calculated from this value was S1%.

Example 9 (1) Introduction of sulfopropyl group An acrylic crosslinked polymer ioy ring-opening modified with ethylene glycol obtained in the same manner as in Example 7-[1] was treated in the same manner as in Example 6-(1). Sulfopropylation was performed at an exchange capacity of 0.4 (6 meq/f-res in
The functional group introduction rate with respect to the reactive monomer units in the polymer calculated from this value was 57%.

Example 10 (1) Production of acrylic crosslinked polymer Polyvinylpyrrolidone -,30/,g? , glycidyl methacrylate 27f, 2-hydroxyethyl methacrylate 3. θ
1 and co, 2'-azobisisobutyronitrile 0.39
Ethanol b 7.9? Polymerization was carried out at 70° C. for 3 hours under a nitrogen gas flow. The polymerization conversion rate in this polymerization was 95%. Furthermore, when the polymer particles obtained by this polymerization were observed with a scanning electron microscope, the particle size was 4 tpm.

It was confirmed that the particles were extremely monodisperse true spherical particles with a standard deviation of 0, b. Using this as a seed polymer, the following operations were performed.

First, 0.36t of 2'-azobisisobutyronitrile
, ethylene glycol dimethacrylate, 76.6 f
, /, 2-dichloroethanda3. g2, sodium dodecyl sulfate 77.66? And 2g01 of water was finely dispersed so that the particle size of oil droplets was 0.5 μm or less. This emulsion was mixed with the monodisperse glycidyl methacrylate-2-hydroxyethyl methacrylate copolymer S, Ot, and sodium dodecyl sulfate θ with a particle size of qμm obtained in the previous polymerization.
.

Next, add 3X polyvinyl alcohol aqueous solution to this, ? 0
．． 3 f was added, and polymerization was carried out at 70° C. for 3 hours under a nitrogen gas flow. What is the polymerization conversion rate? When observed with a scanning electron microscope, the polymer particles obtained by this polymerization were highly monodisperse with a particle size 'IJ prns standard deviation of 3.3%.

Considering the polymerization conversion rate, the polymerization fraction of crosslinkable monomer units in the crosslinked polymer is 6%, and the weight fraction of reactive monomer units derived from glycidyl methacrylate and -monohydroxyethyl methacrylate is /41%, which is The results showed good agreement with the analysis results obtained by pyrolysis gas chromatography.

(2) Ring opening with water The polymer ioy obtained in (1) was mixed with 10% H, So.

Hydrolysis was carried out by reacting in aqueous solution 1072 at a temperature of 50° C. for S hours, and the glycidyl groups in the polymer were ring-opened. After the reaction was completed, the polymer was filtered off and thoroughly washed with demineralized water until the filtrate became neutral, thereby quantitatively obtaining a crosslinked acrylic resin having hydroxyl groups.

(3) Acrylic crosslinked polymer 10 ring-opening modified with water obtained in step (2) of introduction of diethylaminoethyl group
A diethylamine ethyl group was introduced into 9 in the same manner as in Example 3-(3), and the exchange capacity was 0. ! ; meq/t
-res in acrylic anion exchange resin was quantitatively obtained.

The functional group introduction rate with respect to the reactive monomer units in the polymer calculated from this value was j-7X.

Furthermore, when the anion exchange resin produced according to the above formulation was used as a protein separation agent, it showed extremely good separation properties.

Comparative Example/Polymer particles having the same monomer composition and the same average particle size as the acrylic crosslinked polymer particles obtained in Example 3-+1) were produced by suspension polymerization, and the polymer particles were produced by suspension polymerization (yield 20X).
Thereafter, an anion exchange resin was prepared in the same manner as in Example 3-(2) and (3).

The exchange capacity at this time is θ, J meq/r −resi
n, and the functional group introduction rate was -1%. Furthermore, protein separation was performed under the same conditions as in Example 1.
Even though it was classified, it was not possible to measure it due to the increase in pressure drop because of the presence of microparticles.

Comparative example/Example? - has the same monomer composition as the acrylic crosslinked polymer particles obtained in (1), and has an average particle diameter of gJ pm
Polymer particles of Example 7-(2) were prepared by suspension polymerization.
) and (3) to obtain an anion exchange resin.

At this time, the exchange capacity is θ, x3meq/1-resin
The functional group introduction rate was 25X. Furthermore, when protein was separated under the same conditions as in Example 1, non-specific adsorption of the sample to the resin was observed.

From the above Examples and Comparative Examples, the chromatography carrier using the acrylic crosslinked polymer of the present invention has a sufficient amount of functional groups compared to conventional products, and can perform highly sensitive separation without non-specific adsorption of the analyte to the resin. It turns out that it is possible to achieve this.

〔Effect of the invention〕

When the crosslinked polymer particles of the present invention are used as a chromatography carrier, they have the following advantages compared to conventional chromatography carriers.

(a) It has high mechanical strength, allows eluent to flow at high speed, and enables rapid analysis.

Middle) Due to the high degree of crosslinking, the degree of swelling and shrinkage is small and it can be used at high salt concentrations.

(c) Despite the high degree of crosslinking, the amount of functional group introduced into the reactive monomer units constituting the carrier is 0% to 100%, and a sufficient amount of functional groups can be obtained.

) Since the particle size is uniform, the flow rate and the differential pressure relative to it,
The pressure increase is extremely gradual, and particles with the same average particle size can be flowed at a higher speed compared to conventional products.

(e) Since the particle size is uniform, the separation pattern is sharp and the separation ability is high.Compared to conventional products, the number of theoretical plates is higher at the same packing height and the recovery efficiency of the target substance is increased.

(F) Since the particle size is uniform, filling and scale-up are easy.

(g) It is chemically stable over a wide pH range.

[Brief explanation of the drawing]

FIG. 1 is a titration curve, and FIG. 1 is a diagram showing a chromatogram. l...Myoglobinco...Ovalbumin

Claims (2)

[Claims] (1) The weight fraction of crosslinkable monomer units is 60 to 95%, and the weight fraction of acrylic reactive monomer units is 5 to 95%.
40%, with 4 functional groups per reactive monomer unit.
1. Acrylic crosslinked polymer particles characterized in that they have a particle size distribution of 0 to 100%, a standard deviation value of particle size distribution of 10% or less, and a particle size range of 1 to 30 μm. (2) A method for producing acrylic crosslinked polymer particles using the following steps. First step: An acrylic seed polymer is produced by dispersing and polymerizing an acrylic reactive monomer or a crosslinkable monomer and an acrylic reactive monomer in a solvent that dissolves the raw material monomer but does not dissolve the produced polymer. Second step The seed polymer produced in the first step and a crosslinkable monomer in an amount of 1 to 20 times the weight of the seed polymer, or a crosslinkable monomer and an acrylic reactive monomer are seed-polymerized to form a crosslinkable monomer. An acrylic crosslinked raw material polymer is produced in which the weight fraction of units is 60 to 95% and the weight fraction of acrylic reactive monomer units is 5 to 40%. Third Step Acrylic crosslinked polymer particles are manufactured by introducing functional groups into the acrylic crosslinked raw material polymer manufactured in the first step.