JPH11302304A - Production of polymer file particle and production of filler for liquid chromatography - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide both a method capable of readily producing polymer fine particles of uniform particle diameter having a high crosslinking degree, hydrophilic nature, pores of a sufficient size, containing a hydrophilic group densely distributed in the vicinity of the surface of particle and a method for producing a filler for liquid chromatography by using the polymer fine particles. SOLUTION: A hydrophobic crosslinkable monomer (B) or a monomer mixture (D) containing the hydrophobic crosslinkable monomer (B) and a hydrophobic uncrosslinding monomer (C) and an organic solvent (E) are absorbed in hydrophobic polymer fine particles (A) dispersed in an aqueous dispersion medium and polymerized in the presence of a polymerization initiator (F). Further, a hydrophilic monomer (G1) is added in the middle of the polymerization and the polymerization is continued to give hydrophilic polymer fine particles (H1) having a high crosslinking degree and containing pores having 100-1,000 Åaverage pore diameter.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a hydrophilic polymer fine particle having a uniform particle size and a method for producing a packing material for liquid chromatography using the polymer fine particle. Average pore radius is 100-1000Å
The present invention relates to a method for producing hydrophilic polymer fine particles having fine pores and a method for producing a filler for liquid chromatography comprising the hydrophilic polymer fine particles.

[0002]

2. Description of the Related Art Conventionally, micron size (0.5 to 100)
Polymer fine particles having a diameter of about μm) are widely used as organic pigments, toner particles, spacers, separation materials, biochemical carriers, standard particles, cosmetic fillers, various additives or compounding agents, and the like. In polymer fine particles used for this type of application, it is strongly required that the particle diameter be as uniform as possible.

[0003] In some applications, it is required that the polymer fine particles have a hydrophilic functional group or that they have excellent pressure resistance. For example, in a polymer fine particle used as a separation material, a material that separates various substances can be obtained by retaining an ion exchange group or a hydrophilic hydroxyl group. In particular, a packing material for liquid chromatography has been required to have the following properties as a packing material. That is, the particles are true spherical particles, have an average particle size of about 1 to 20 μm in analytical applications, and have a uniform particle size distribution. In addition, it has been required to have a large pressure resistance, that is, a high degree of crosslinking so that column packing and analysis under high pressure are possible.
In particular, in the analysis in an aqueous system, it is also required to have sufficient hydrophilicity. In addition, in the case of polymer fine particles used as a carrier, a hydrophilic functional group near the surface of the polymer fine particles can be used to react with a chemical substance or a biological substance.

[0004] As a conventional method for producing micron-sized polymer fine particles, various methods using suspension polymerization, emulsion polymerization or dispersion polymerization are known. However, when the suspension polymerization method is used, a complicated classification operation is required in order to make the particle diameter uniform since the obtained polymer fine particles have a wide particle size distribution. Further, it is very difficult to classify to the monodispersion level by the current classification method.

[0005] When the emulsion polymerization method is used, although the particle size of the obtained polymer fine particles is uniform, it has been very difficult to increase the average particle size to the micron size. Further, when the dispersion polymerization method is used, the type of the obtained polymer fine particles is difficult because the crosslinking reaction is difficult or the copolymerization of the hydrophilic monomer and the hydrophobic monomer is difficult. Had restrictions. In addition, since a large amount of organic solvent is required for the dispersion medium, there is a difficulty in handling during production.

As a solution to the above-mentioned problems, polymer particles are further absorbed and polymerized by polymer fine particles obtained by emulsion polymerization or dispersion polymerization, and the particle size is maintained while maintaining the uniformity of the particle size. Methods for increasing are known.

[0007] In the method of further polymerizing a polymer particle by absorbing the monomer, in order to obtain a polymer particle having a high degree of crosslinking, a monomer mixture containing a crosslinkable monomer is added to the polymer particle. It is known that the polymer may be absorbed and polymerized (Japanese Patent Publication No. 5-62605).

Further, as a method for imparting hydrophilicity to the polymer fine particles simultaneously with the crosslinking reaction, there is known a method in which a mixture of a hydrophilic monomer and a crosslinkable monomer is absorbed into the polymer fine particles and polymerized. (For example, Japanese Patent Publication No. 7-103206)
Issue publication).

However, in these methods, since the crosslinked particles obtained after the polymerization contain hydrophilic groups derived from the hydrophilic monomer, the pressure resistance is reduced,
The reaction to changes in the external environment (such as pH and salt concentration) is slowed down by the presence of hydrophilic groups inside the polymer microparticles, and swelling and shrinkage are likely to occur. Because of its large size, there is a problem that it is difficult to sufficiently absorb the polymer particles.

[0010] On the other hand, Japanese Patent Application Laid-Open No. 61-215603 discloses that after a hydrophobic monomer is first absorbed by polymer fine particles,
It is shown that by using a method of absorbing a hydrophilic monomer, that is, a two-stage absorption method, the hydrophilic monomer can be efficiently absorbed by the polymer fine particles. JP 61
Although it is said that the above-mentioned problem that the hydrophilic monomer is hardly absorbed by the polymer fine particles is improved by using the production method described in -215603, the problems described above are still solved. I didn't.

Further, attempts have been made to prepare polymer fine particles suitable for a packing material for liquid chromatography by applying this polymerization method. For example, polymer particles are firstly polymerized by absorbing a monomer mixture containing no hydrophilic monomer and mainly containing a crosslinkable monomer, and adding the hydrophilic monomer during the polymerization reaction. A method has been proposed (JP-A-6-262070). However, in this method, since the pore radius is limited to 10 to 75 °, the low molecular weight compound is separated from the mixture of the high molecular weight compound and the low molecular weight compound in a reverse phase mode without performing a pretreatment operation. Agents. Therefore, it cannot be applied to the ion exchange mode in which the functional group on the inner surface of the pore needs to act on the polymer compound, nor to the gel permeation chromatography using the sieving principle.

Japanese Patent Publication No. 8-7197 discloses a process for producing a packing material for liquid chromatography comprising particles having a high degree of crosslinking, being porous, and having hydrophilic groups unevenly distributed near the surface. . However, in this method, since suspension polymerization is used, it is difficult to make the particle size distribution uniform, and a complicated classification operation has to be performed.

As described above, according to the conventional method for producing a packing material for liquid chromatography, it is a true spherical polymer particle having an average particle size of a micron size and a uniform particle size distribution, and has a high degree of crosslinking and hydrophilicity. In addition, it was not possible to easily obtain a filler which sufficiently satisfies various conditions that the hydrophilic group is unevenly distributed near the particle surface and has pores of a sufficient size.

[0014]

SUMMARY OF THE INVENTION An object of the present invention is to eliminate the drawbacks of the conventional method for producing fine polymer particles, to have a high degree of crosslinking, to be hydrophilic, and to have pores of a sufficient size. Method for easily producing polymer fine particles having a uniform particle diameter in which hydrophilic groups are unevenly distributed near the particle surface, and a filler for liquid chromatography using such polymer fine particles It is to provide a manufacturing method of.

[0015]

According to the first aspect of the present invention, a hydrophobic polymer fine particle (A) dispersed in an aqueous dispersion medium is provided.
A monomer mixture (D) containing a hydrophobic crosslinkable monomer (B) or a hydrophobic crosslinkable monomer (B) and a hydrophobic non-crosslinkable monomer (C), and an organic solvent (E). ) Is absorbed, polymerized in the presence of the polymerization initiator (F), and further added with a hydrophilic monomer (G1) in the course of the polymerization to continue the polymerization, whereby the crosslinked and average fine particles are formed. Hole radius is 100-10
This is a method for producing polymer fine particles, which comprises producing hydrophilic polymer fine particles (H1) having pores of 00 °.

Further, according to the present invention, the hydrophobic polymer fine particles (A) dispersed in an aqueous dispersion medium are provided with a hydrophobic crosslinkable monomer (B) or a hydrophobic crosslinkable monomer (A). B) and a monomer mixture (D) containing a hydrophobic non-crosslinkable monomer (C) and an organic solvent (E) are absorbed, and polymerized in the presence of a polymerization initiator (F), and further polymerized. During the course of the polymerization, a monomer (G2) having a functional group (J) that can be converted to a hydrophilic group (I) by a chemical reaction is added to continue polymerization, and after polymerization, the functional group (J) is converted into a hydrophilic group (I). By converting to the group (I), it is crosslinked and has an average pore radius of 100 to 10
A method for producing polymer microparticles, which comprises producing hydrophilic polymer microparticles (H2) having pores of 00 °.

Further, according to the present invention, the hydrophobic polymer fine particles (A) dispersed in the aqueous dispersion medium are provided with the hydrophobic crosslinkable monomer (B) or the hydrophobic crosslinkable monomer (A). B) and a monomer mixture (D) containing a hydrophobic non-crosslinkable monomer (C) and an organic solvent (E) are absorbed, and polymerized in the presence of a polymerization initiator (F), and further polymerized. By adding the hydrophilic monomer (G1) in the middle of the reaction and continuing the polymerization,
Crosslinked and having an average pore radius of 100-1000 °
This is a method for producing a packing material for liquid chromatography comprising hydrophilic polymer fine particles (H1) having fine pores.

In the third aspect of the present invention, preferably, as described in the fourth aspect, the hydrophilic monomer (G
As 1), a monomer having an ion exchange group is used, and the hydrophilic polymer fine particles (H1) are used as a filler for ion exchange.

The invention according to claim 5 is characterized in that the hydrophobic polymer fine particles (A) dispersed in the aqueous dispersion medium are added to the hydrophobic crosslinkable monomer (B) or the hydrophobic crosslinkable monomer (A). B) and a monomer mixture (D) containing a hydrophobic non-crosslinkable monomer (C) and an organic solvent (E) are absorbed, and polymerized in the presence of a polymerization initiator (F), and further polymerized. During the course of the polymerization, a monomer (G2) having a functional group (J) that can be converted to a hydrophilic group (I) by a chemical reaction is added to continue polymerization, and after polymerization, the functional group (J) is converted into a hydrophilic group (I). By converting to the group (I), it is crosslinked and has an average pore radius of 100 to 10
This is a method for producing a packing material for liquid chromatography comprising hydrophilic polymer fine particles (H2) having pores of 00 °.

In the fifth aspect of the present invention, preferably, as in the sixth aspect, the hydrophilic group (I) is an ion exchange group, and the hydrophilic polymer fine particles (H2)
Is a filler for ion exchange.

Hereinafter, the present invention will be described in detail. (Aqueous dispersion medium) In the present invention, as the aqueous dispersion medium, water, an appropriate organic solvent soluble in water, and a mixture thereof can be used. Examples of the organic solvent include alcohols such as methanol, ethanol, and isopropanol; ethyl acetate; acetone; acetonitrile; and the like, but are not particularly limited thereto. However, it is necessary that the aqueous dispersion medium does not dissolve the hydrophobic polymer fine particles (A).

If necessary, a dispersing aid for dispersing the hydrophobic polymer fine particles (A) may be added to the aqueous dispersion medium. Examples of such a dispersing aid include, but are not particularly limited to, high molecular compounds such as polyvinyl alcohol, polyvinyl pyrrolidone, gelatin, starch, hydroxycellulose, and polyvinyl ether; sodium lauryl sulfate, sodium lauryl benzene sulfonate, Examples thereof include ionic surfactants such as sodium oxyethylene lauryl sulfate and sodium dialkyl sulfosuccinate; and nonionic surfactants such as polyoxyethylene nonyl phenyl ether, polyethylene glycol monostearate, and sorbitan monostearate. .

The amount of the aqueous dispersion medium to be used is preferably 500 to 200,000 parts by weight, more preferably 100 to 100 parts by weight of the hydrophobic polymer fine particles (A) described below.
It is 1000 to 100,000 parts by weight. If the amount of the aqueous dispersion medium is less than 500 parts by weight, it may be difficult to uniformly disperse the hydrophobic polymer fine particles (A) in the aqueous dispersion medium. The efficiency may be reduced and the particle size may not be sufficient.

(Hydrophobic Polymer Fine Particles (A)) The hydrophobic polymer fine particles (A) are not particularly limited, but can be used to obtain micron-sized polymer fine particles (H1) or (H2). And preferably an average particle size of 0.1 to 3
0 μm, more preferably 0.5 to 10 μm is used. If the average particle size is less than 0.1 μm, large micron-sized polymer fine particles (H1) or (H2) may not be obtained, and if it exceeds 10 μm, the uniformity of the particle size may be impaired.

Further, the CV value (%) of the particle diameter of the hydrophobic polymer fine particles (A) / (standard deviation / average particle diameter) ×
100 ° is preferably 20% or less, more preferably 1%.
5% or less. If the CV value exceeds 20%, the resulting polymer fine particles (H1) or (H2) may have large variations in particle size.

The hydrophobic polymer fine particles (A) are preferably swellable and capable of absorbing a sufficient amount of the hydrophobic crosslinkable monomer (B) or monomer mixture (D) described below. Non-crosslinkable or low crosslinkable polymers are used. Here, the low crosslinkability is obtained by polymerizing a monomer mixture obtained by mixing a crosslinkable monomer in a ratio of 0 to 10% by weight and a non-crosslinkable monomer in a ratio of 100 to 90% by weight. It refers to a polymer. When the hydrophobic polymer fine particles (A) have a high degree of crosslinking,
This is because the monomer (G1) or (G2) to be added later becomes difficult to be absorbed.

The method for producing the hydrophobic polymer fine particles (A) is not particularly limited, and can be obtained by polymerizing a monomer by known suspension polymerization, emulsion polymerization, dispersion polymerization, or the like.

Examples of the monomer (K) used to obtain the hydrophobic polymer fine particles (A) include styrene, α-
Styrene derivatives such as methylstyrene, p-methylstyrene, chloromethylstyrene, styrenesulfonic acid, divinylbenzene, divinyltoluene, divinylxylene, divinylethylbenzene, divinylnaphthalene; vinyl chloride, vinyl acetate, vinyl propionate, vinyl stearate, etc. Vinyl esters; methyl (meth) acrylate, ethyl (meth) acrylate, butyl (meth) acrylate, 2-ethylhexyl (meth) acrylate, stearyl (meth) acrylate, ethylene glycol di (meth) acrylate, Polyethylene glycol di (meth)
Acrylate, 1,3-butylene glycol di (meth)
Acrylate, 1,6-hexane glycol di (meth)
Acrylate, neopentyl glycol di (meth) acrylate, polypropylene glycol di (meth) acrylate, trimethylolethanetri (meth) acrylate, trimethylolpropanetri (meth) acrylate, tetramethylolpropanetri (meth) acrylate, tetramethylolmethanetetra (Meth) acrylate derivatives such as (meth) acrylate; 1,3-
Aliphatic diene compounds such as butadiene, isoprene and 1,4-hexanediene; and derivatives of the above monomers.

It is preferable to use a monomer having no ion exchange group or the like as the above-mentioned monomer. In the first and third aspects of the present invention, the polymer particles (H1) finally obtained have hydrophilicity because most of the hydrophilic functional groups of the polymer particles (H1) are described later. This is because it is derived from the hydrophilic monomer (G1). In the second and fifth aspects of the present invention, the polymer fine particles (H2) finally obtained are hydrophilic because most of the hydrophilic functional groups of the polymer fine particles (H2) are described later. This is because the monomer (G2) is derived from the converted hydrophilic group (I).

(Hydrophobic Crosslinkable Monomer (B)) As the hydrophobic crosslinkable monomer, the monomer (K) used for constituting the above-mentioned hydrophobic polymer fine particles (A) can be used. Among them, a crosslinkable monomer (a monomer having two or more functionalities) is used.
However, the crosslinkable monomer used to form the hydrophobic polymer fine particles (A) and the hydrophobic crosslinkable monomer (B) may be the same or different. .

(Hydrophobic non-crosslinkable monomer (C)) As the above-mentioned hydrophobic non-crosslinkable monomer (C), a monomer (A) for constituting the above-mentioned hydrophobic polymer fine particles (A) may be used. One or more of the non-crosslinkable monomers of K) are used. However, the non-crosslinkable monomer used to constitute the hydrophobic polymer fine particles (A) and the hydrophobic non-crosslinkable monomer (C) are:
They may be the same or different.

(Monomer Mixture (D)) The mixing ratio of the hydrophobic crosslinkable monomer (B) and the hydrophobic noncrosslinkable monomer (C) is not particularly limited. ,Preferably,
80% by weight or more of the mixture of the hydrophobic crosslinking monomer (B),
More preferably, the content is 90% by weight or more. By adjusting the mixing ratio of the hydrophobic crosslinkable monomer (B) to 80% by weight or more, the polymer particles (H1) or (H
It is easy to increase the degree of crosslinking in 2) and increase the pressure resistance. In order to increase the pressure resistance, it is desirable to use 100% of the hydrophobic crosslinkable monomer (B). Purification is required. Therefore, considering the limitations of the purification technique and the complexity of the operation, sufficient pressure resistance can be obtained without necessarily using only the hydrophobic crosslinkable monomer (B). It is desirable to use a monomer mixture (D) containing 80% by weight or more of the monomer (B).

(Amount of Hydrophobic Crosslinkable Monomer (B) or Monomer Mixture (D)) The above hydrophobic crosslinkable monomer (B)
Alternatively, the amount of the monomer mixture (D) used is determined by the difference between the particle size of the hydrophobic polymer fine particles (A) and the particle size of the polymer fine particles (H1) or (H2) as the product. Although not particularly limited, preferably, the hydrophobic crosslinkable monomer (B) or the monomer mixture (D) is 100 to 3 parts by weight per 100 parts by weight of the hydrophobic polymer fine particles (A).
0000 parts by weight, more preferably 200 to 20,000 parts by weight. When the use ratio of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D) is less than 100 parts by weight, it becomes difficult to increase the degree of crosslinking of the polymer fine particles (H1) or (H2). In some cases, the polymer fine particles (H1) or (H2) having sufficient pressure resistance cannot be obtained. If the amount exceeds 30,000 parts by weight, the hydrophobic crosslinkable monomer (B) or monomer mixture (D ) Becomes difficult to be absorbed by the polymer fine particles (A), and fine particles may be generated after polymerization.

The method of adding the hydrophobic crosslinkable monomer (B) or the monomer mixture (D) is not particularly limited, either. The polymerization initiator (F) may be dissolved in the monomer mixture (D) and then dispersed in the dispersion medium, or may be dispersed in the dispersion medium separately from the polymerization initiator (F) and then mixed. Good.

(Absorption of Hydrophobic Crosslinkable Monomer (B) or Monomer Mixture (D) into Polymer Fine Particles (A)) The hydrophobic crosslinkable monomer is added to the hydrophobic polymer fine particles (A). As a method for absorbing the body (B) or the monomer mixture (D), any known method can be used. For example, after forming the hydrophobic crosslinkable monomer (B) or the monomer mixture (D) into finer droplets than the hydrophobic polymer fine particles (A), the hydrophobic crosslinkable monomer (B) or A method in which the monomer mixture (D) and the hydrophobic polymer fine particles (A) are mixed and stirred (JLJan
sson, MCWellons, GWPoehlein: J Polym Sci Part
C; Vol. 21 No. 11 pp 937-943) or a method using a swelling aid (Japanese Patent Publication No. 57-24369).

(Organic Solvent (E)) As the above organic solvent (E), those which dissolve the monomers (B) and (D) but do not dissolve the polymer thereof are selected. The amount of the organic solvent (E) used is preferably 20 to 2,000 parts by weight based on 100 parts by weight of the monomer (B) or (D). If the amount of the organic solvent (E) is less than 20 parts by weight, the pore diameter becomes too small, and the functional group on the inner surface of the pore cannot sufficiently interact with the sample. If the amount exceeds 2,000 parts by weight, the pore diameter becomes too large, and the pressure resistance may decrease, or aggregates may be generated during the polymerization. The organic solvent (E) is preferably added to the reaction system after being dissolved in the monomer (B) or the monomer mixture (D). Specific examples of the organic solvent (E) include alcohols such as methanol, ethanol, propanol and isoamyl alcohol; aliphatic organic solvents such as ethyl acetate, acetone and methyl ethyl ketone; aromatics such as benzene, toluene and xylene. Examples thereof include hydrocarbons.

(Polymerization Initiator (F)) The polymerization initiator (F) is not particularly limited, and a known water-soluble or oil-soluble radical polymerization initiator can be used. The amount of the polymerization initiator (F) used is the amount of the hydrophobic polymer fine particles (A).
It is preferable that the amount be 1 to 2000 parts by weight based on 100 parts by weight. When the amount of the polymerization initiator (F) used is less than 1 part by weight, the polymerization reaction may be insufficient or the polymerization may take a long time. May generate aggregates.

The polymerization initiator (F) may be added to the reaction system after being dissolved in the hydrophobic crosslinkable monomer (B) or the monomer mixture (D). It may be added to the reaction system as a solution or dispersion dissolved in another liquid substance.

Specific examples of the above polymerization initiator (F) include persulfates such as potassium persulfate, sodium persulfate and ammonium persulfate; cumene hydroperoxide, benzoyl peroxide, lauroyl peroxide, octanoyl peroxide. Oxide, o-chlorobenzoyl peroxide, acetyl peroxide, t-butyl hydroperoxide, t-butyl peroxyacetate, t-butyl peroxyisobutyrate, 3,
5,5-trimethylhexanoyl peroxide, t-
Butyl peroxy-2-ethylhexanoate, di-t
Organic peroxides such as butyl peroxide; 2,2 ′
-Azobisisobutyronitrile, 2,2'-azobis (2,4-dimethylvaleronitrile), 4,4'-azobis (4-cyanopentanoic acid), 2,2'-azobis (2-methylbutyro Azo compounds such as nitrile) and azobiscyclohexanecarbonitrile; and derivatives of the above-mentioned polymerization initiators.

(Polymerization) A system comprising a hydrophobic polymer fine particle (A), a hydrophobic crosslinkable monomer (B) or a monomer mixture (D), a polymerization initiator (F) and an organic solvent (E) is used. By raising the temperature to a temperature at which the polymerization initiator (F) generates radicals, polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D) is performed. The polymerization temperature varies depending on the type of the polymerization initiator (F), but is usually 40 to 120.
° C.

In order to prevent aggregation of the particles during the polymerization, an aqueous solution containing the above-mentioned dispersing aid may be added to the reaction system. The dispersion medium composition in this case was used when dispersing the hydrophobic polymer fine particles (A), the hydrophobic crosslinkable monomer (B) or the monomer mixture (D), the polymerization initiator (F), and the like. It may be different from the dispersion medium.

Up to this stage of the polymerization, the inventions according to claims 1 and 3 and the inventions according to claims 2 and 5 are the same, but the subsequent operations are different, so that they will be described separately. .

First, the first and third aspects of the present invention will be described.

According to the first and third aspects of the present invention, a hydrophilic monomer (G1) is added to the reaction system during the polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D). I do. The term “during polymerization” means before the polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D) is completely completed, and preferably the hydrophobic crosslinkable monomer (B) or When the polymerization rate of the monomer mixture (D) is in the range of 20 to 98%, more preferably 30 to 95%, and the polymerization initiator (F) added first remains, The monomer (G1) is added.

Further, after the polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D) is started, that is, when the temperature reaches a predetermined temperature, the addition of the hydrophilic monomer (G1) is continued. Is preferably 0.3 to 20 hours, more preferably 0.5 to 10 hours. If the time is less than 0.3 hours, the hydrophilic monomer (G1) is added at a stage where the polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D) has not sufficiently proceeded. It becomes difficult to unevenly distribute the hydrophilic monomer in the vicinity of the surface of the finally obtained polymer fine particles, and if it exceeds 20 hours, the polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D) may occur. The process is nearly completed, and the polymerization of the hydrophilic monomer (G1) becomes difficult. When the hydrophilic monomer (G1) is added, the polymerization initiator (F) may be further added.

(Hydrophilic monomer (G1)) The hydrophilic monomer (G1) is added to introduce a hydrophilic group near the surface of the finally obtained hydrophilic polymer fine particles (H1). Is done. The amount of the hydrophilic monomer (G1) to be added is preferably 100 to 50,000 parts by weight, more preferably 500 to 100 parts by weight of the hydrophobic polymer fine particles (A).
20,000 parts by weight. When the addition ratio of the hydrophilic monomer (G1) is less than 100 parts by weight, the hydrophilicity near the surface of the obtained hydrophilic polymer fine particles (H1) may be insufficient, and 50,000 parts by weight may be used. If it exceeds, aggregates may be generated.

The hydrophilic monomer (G1) tends to cause aggregation during the polymerization reaction. Therefore, the hydrophilic monomer (G1)
It is preferable to stop the polymerization reaction in a state where the polymerization does not proceed sufficiently (that is, before aggregation occurs).

It is not necessary to polymerize all of the hydrophilic monomer (G1), including the case where the reaction is stopped before aggregation, as described above. The reaction can be stopped when the required degree of hydrophilicity is obtained. In this case, the amount of the hydrophilic monomer (G1) added depends on the amount of the hydrophilic polymer fine particles (H
It will not match the amount of the hydrophilic monomer (G1) component contained in 1). In addition, unreacted hydrophilic monomer (G1)
Can be removed by washing after polymerization.

Since the presence of an organic solvent in the polymerization reaction system increases the probability of generation of aggregates during the polymerization of the hydrophilic monomer (G1), the combination of the monomer type and the organic solvent type used must be carefully selected. You need to decide.

The polymerization time of the hydrophilic monomer (G1) is preferably from 0.3 to 40 hours, more preferably from 0.5 to 20 hours from the start of the addition. When the polymerization time of the hydrophilic monomer (G1) is less than 0.3 hours, the hydrophilic monomer (G1)
If the polymerization rate of 1) is insufficient, and if it exceeds 40 hours, agglomerates are likely to be generated.

The hydrophilic monomer (G1) is not particularly limited as long as it has a hydrophilic group, and examples thereof include (meth) acrylic acid, itaconic acid, crotonic acid, Allylsulfonic acid, maleic acid, fumaric acid,
Cation exchange groups such as sodium styrenesulfonate, potassium styrenesulfonate, 2-acrylamido-2-methylpropanesulfonic acid, 3-sulfopropyl acrylate, 3-sulfopropyl methacrylate, and 2- (meth) (acryloyloxyethyl) acid phosphate A monomer having an anion exchange group such as dimethylaminoethyl (meth) acrylate, diethylaminoethyl (meth) acrylate, allylamine, (meth) acrylate hydroxypropyltrimethylammonium chloride; (meth) acrylamide, N
-Methylacrylamide, N-ethylacrylamide,
Acid amides such as N-methylmethacrylamide, N-ethylmethacrylamide, N, N-dimethylacrylamide; ethylene glycol mono (meth) acrylate;
-Hydroxyethyl (meth) acrylate, glycidyl (meth) acrylate, (meth) acrylonitrile; and derivatives of the above monomers.

The method of adding the hydrophilic monomer (G1) is as follows. During the polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D), the hydrophilic monomer (G1) is added. Can be dropped from the dropping funnel into the reaction system.

(Hydrophilic Polymer Fine Particles (H1)) The average particle diameter of the hydrophilic polymer fine particles (H1) obtained by the invention of claims 1, 3 and 4 is preferably about 0.5-1.
00 μm, and has a uniform particle size distribution. The uniform particle size distribution is defined as CV value (%) ｛(standard deviation / average particle size).
× 100 ° preferably means 10% or less.

The pore radius is limited to 100-1000 °. If the pore radius is less than 100 °, the sample is difficult to diffuse into the pores when the sample is a polymer, the functional groups on the pore inner surface do not act effectively, and the pore radius is less than 10 °.
If it exceeds 00 °, a problem occurs in the pressure resistance.

The hydrophilic polymer fine particles (H1)
Are crosslinked to a high degree of crosslinking. This is because the monomer mixture (D) contains a large amount of the hydrophobic crosslinkable monomer (B), and as described above, the hydrophobic crosslinkable monomer (B) By increasing the content, polymer particles (H1) having a high degree of crosslinking can be obtained.

Since the hydrophilic monomer (G1) is added and polymerized, the hydrophilic polymer fine particles (H1) have hydrophilicity. In particular, the hydrophilic monomer (G1) is added and polymerized during the polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D). Since (A), the hydrophobic crosslinkable monomer (B) and the hydrophobic non-crosslinkable monomer (C) have few hydrophilic groups, the finally obtained hydrophilic group is a hydrophilic polymer. The particles are unevenly distributed near the surface of the fine particles (H1), and the inside of the hydrophilic high-molecular fine particles (H1) has few hydrophilic groups. Therefore, the hydrophilic polymer particles (H1) have sufficient pressure resistance, though they have hydrophilicity due to little swelling and shrinkage. The hydrophilic polymer fine particles (H1) have substantially effective pores inside or on the surface of the particles. It can interact.

Next, the second and fifth aspects of the invention will be described.

According to the second and fifth aspects of the present invention,
During the polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D) in the description of the above polymerization section,
A monomer (G2) having a functional group (J) that can be converted to a hydrophilic group (I) by a chemical reaction is added to the reaction system. The term “during polymerization” means before the polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D) is completely completed, and preferably the hydrophobic crosslinkable monomer (B) or The degree of polymerization of the monomer mixture (D) is from 20 to 98%, more preferably from 30 to 98%.
The above monomer (G2) is added in a state where the content of the polymerization initiator (F) is in the range of about 95% and the polymerization initiator (F) initially added remains.

Further, after the polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D) is started, that is, after the temperature reaches a predetermined temperature, the polymerization is continued until the addition of the monomer (G2). The time is preferably from 0.3 to 20 hours, more preferably from 0.5 to 10 hours. In the case where the polymerization time is less than 0.3 hours, the polymerization of the monomer (G) is not carried out at a stage where the polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D) has not sufficiently proceeded.
Due to the addition of 2), the monomer (G2) is less likely to be unevenly distributed near the surface of the finally obtained polymer fine particles, and if it exceeds 20 hours, the hydrophobic crosslinkable monomer (B) or the monomer The polymerization of the mixture (D) is almost complete, and the monomer (G2)
Polymerization hardly occurs. When the monomer (G2) is added, the polymerization initiator (F) may be further added.

(Monomer (G2) Having Functional Group (J) That Can Be Converted to Hydrophilic Group (I) by Chemical Reaction) The monomer (G2) is obtained by using hydrophilic polymer fine particles finally obtained. It is added to introduce a hydrophilic group near the surface of (H2). The amount of the monomer (G2) to be added is preferably 1 to 100 parts by weight of the hydrophobic polymer fine particles (A).
00 to 50,000 parts by weight, more preferably 500 to 20 parts by weight
000 parts by weight. When the addition ratio of the monomer (G2) is 10
If the amount is less than 0 parts by weight, the hydrophilicity near the surface of the obtained hydrophilic polymer fine particles (H2) may be insufficient, and if it exceeds 50,000 parts by weight, aggregates may be generated.

Since the monomer (G2) easily causes aggregation during the polymerization reaction, the polymerization reaction may be stopped in a state where the polymerization of the monomer (G2) does not proceed sufficiently (that is, before the aggregation occurs). preferable.

It is not necessary to polymerize all of the monomers (G2), including the case where the reaction is stopped before aggregation. The reaction can be stopped when the required amount of functional group (J) is obtained. In this case, the amount of the monomer (G2) added does not match the amount of the monomer (G2) component contained in the hydrophilic polymer fine particles (H2) obtained after the polymerization. The unreacted monomer (G2) can be removed by washing after polymerization.

Since the presence of an organic solvent in the polymerization reaction system increases the probability that an aggregate will be generated during the polymerization of the monomer (G2), the combination of the monomer type and the organic solvent type to be used is carefully determined. There is a need.

The polymerization time of the monomer (G2) is preferably from 0.3 to 40 hours from the start of the addition, more preferably from 0.5 to 40 hours.
~ 20 hours. The polymerization time of the monomer (G2) is 0.3
When the time is less than the time, the polymerization rate of the monomer (G2) is insufficient, and when the time is more than 40 hours, an aggregate is easily generated.

The monomer (G2) is a monomer having a functional group (J) that can be converted into a hydrophilic group (I) by a chemical reaction. The chemical reaction referred to here is a reaction (hydrolysis reaction or the like) in which the functional group (J) is converted into a hydrophilic group (I) by decomposing in some form, or a compound having the hydrophilic group (I) is functionalized. Reaction to be bonded to group (J) (condensation reaction,
Esterification reaction, acylation reaction, etc.).

Examples of the functional group (J) which can be converted into the hydrophilic group (I) include various ester groups, amide groups,
Examples include a mercapto group, an amino group, a carbonyl group, a hydroxyl group, an epoxy group, and a benzyl group.

Specific examples of the monomer (G2) include:
Organic acids such as (meth) acrylic acid, itaconic acid, fumaric acid, and maleic acid; dimethylaminoethyl (meth) acrylate, diethylaminoethyl (meth) acrylate, methyl (meth) acrylate, ethyl (meth) acrylate, 2-ethylhexyl ( (Meth) acrylates such as (meth) acrylate; (meth) acrylamide, (meth) acrylonitrile, 2-hydroxyethyl (meth) acrylate, (poly) ethylene glycol mono (meth) acrylate, (poly) propylene glycol mono ( (Meth) acrylic acid derivatives such as meth) acrylate and glycidyl (meth) acrylate; styrene derivatives such as styrene and α-methylstyrene; allylamine and vinyl acetate.

As for the method of adding the monomer (G2), the monomer (G2) may be dropped during the polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D). From the reaction system.

(Hydrophilization of Functional Group (J) by Chemical Reaction) The functional group (J) of the monomer (G2) is converted into a hydrophilic group (I) by the above chemical reaction. The hydrophilic group (I) is an ion exchange group such as a carboxyalkyl group, a sulfonic acid group, or various amino groups, or a hydrophilic hydroxyl group. Thus, the polymer fine particles (H2) in the final form can be made hydrophilic fine particles.

The chemical reaction is performed, for example, as follows. When methyl methacrylate is used as the monomer (G2), the functional group (J) is a methyl ester group, and the ester group is hydrolyzed by heating under alkalinity to form a hydrophilic group (I). It is converted to a certain carboxyl group. Alternatively, when glycidyl methacrylate is used as the monomer (G2), the glycidyl group as the functional group (J) is converted to the sulfonic acid as the hydrophilic group (I) by reacting with a nitrite compound under acidic conditions. Is converted to the base. Selection of a chemical reaction for the functional group (J) of the monomer (G2) can be performed from known techniques. However, the chemical reaction needs to be performed under conditions that do not damage the polymer microparticles (H2).

(Hydrophilic Polymer Fine Particles (H2)) The average particle diameter of the hydrophilic polymer fine particles (H2) obtained by the invention of claims 2, 5 and 6 is preferably about 0.5 to 1
00 μm, and has a uniform particle size distribution. The uniform particle size distribution is defined as CV value (%) ｛(standard deviation / average particle size).
× 100 ° preferably means 10% or less.

The pore radius is limited to 100-1000 °. If the pore radius is less than 100 °, the sample is difficult to diffuse into the pores when the sample is a polymer, the functional groups on the pore inner surface do not act effectively, and the pore radius is less than 10 °.
If it exceeds 00 °, a problem occurs in the pressure resistance.

Further, the hydrophilic polymer fine particles (H2)
Are crosslinked to a high degree of crosslinking. This is because the monomer mixture (D) contains a large amount of the hydrophobic crosslinkable monomer (B), and as described above, the hydrophobic crosslinkable monomer (B) By increasing the content, polymer particles (H2) having a high degree of crosslinking can be obtained.

Further, since the monomer (G2) is added and polymerized, and the functional group (J) of the monomer (G2) is converted into the hydrophilic group (I), the polymer fine particles ( H2) has hydrophilicity. Particularly, since the monomer (G2) is added and polymerized during the polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D), the hydrophobic crosslinkable polymer (A), Since the hydrophobic crosslinkable monomer (B) and the hydrophobic non-crosslinkable monomer (C) have few hydrophilic groups, the hydrophilic groups finally obtained are on the surface of the polymer fine particles (H2). And the number of hydrophilic groups inside the particles is reduced. Therefore, the hydrophilic polymer particles (H2) have sufficient pressure resistance, though they have hydrophilicity because of little swelling and shrinkage. The hydrophilic polymer fine particles (H2) have substantially effective pores inside or on the surface of the particles. It can interact.

(The hydrophilic polymer fine particles (H1) obtained by the method according to the first aspect and the hydrophilic polymer fine particles (H2) obtained by the method according to the second aspect) Application) hydrophilic polymer fine particles (H1) or (H
The application of 2) is not particularly limited, and can be used in various fields. For example, by using the following additional method, it can be used in various fields as described in the prior art.

That is, the polymer fine particles (H1) or (H1)
2) Including ferromagnetic metals such as iron, cobalt or nickel; magnetic materials such as alloys and compounds such as magnetite, hematite and ferrite, or inorganic pigments, carbon black, and various dyes, It can be suitably used for toner particles and coloring materials. In addition, as dyes, for example, Balifast Red, Opla
s Red, Oil Scarlet (Orient Chemical), Oil Bl
ue V, Oil Green (manufactured by Daiwa Kako), Sumiplast Blue,
Oil dyes such as Sumiplast Yellow (manufactured by Sumitomo Chemical); O
rient Oil Blue (manufactured by Orient Kako), Sumilaron Br
illiant Blue, Sumikaron Violet (Sumitomo Chemical), Ka
Disperse dyes such as yalon Polyester Blue and Kayalon Polyester Red (manufactured by Nippon Kayaku) can be exemplified.

Further, the obtained hydrophilic polymer fine particles (H
A carrier for an immunoreagent by adding and immobilizing various immunologically active substances (rheumatic factor, human albumin, immunoglobulin, streptolysin O, C-reactive protein, alpha fetoprotein, etc.) to 1) or (H2). It is also possible to use Further, by immobilizing enzymes, the enzyme can be used as an immobilized enzyme.

(Production of the packing material for liquid chromatography according to the third aspect of the present invention) The packing material for liquid chromatography according to the third aspect of the present invention is as described in the first aspect.
The hydrophilic polymer fine particles (H1) having a high degree of crosslinking and having effective pores obtained by the production method according to the invention described in (1) above are used as a filler for liquid chromatography. Therefore, the steps up to obtaining the hydrophilic polymer fine particles (H1) are performed in the same manner as the above-described method for producing the hydrophilic polymer fine particles (H1).

Further, by using a monomer having an ion-exchange group as the hydrophilic monomer (G1), the hydrophilic polymer fine particles (H1) can be charged for ion exchange. And a packing material for liquid chromatography. In this case, examples of the hydrophilic monomer (G1) include the above-described monomers having a cation exchange group or an anion exchange group.

When used as a filler for liquid chromatography, a column for liquid chromatography can be constituted by packing the above hydrophilic polymer fine particles (H1) into a column made of stainless steel or the like. Upon filling, an appropriate method can be used, and particularly, a wet method (slurry method) is preferably used. That is, the hydrophilic polymer fine particles (H1) can be filled by dispersing a predetermined amount in a dispersion medium such as a solvent used as an eluent, and then press-fitting the column with a packer or the like.

When the hydrophilic polymer fine particles (H1) are used as a filler for liquid chromatography, the hydrophilic monomer (G1) can be appropriately selected to be applicable to various separation modes of liquid chromatography. Can be. For example, in the case of ion exchange chromatography, a hydrophilic monomer (G1) having an ion exchange group may be used.
A monomer having a hydrophobic group or the like may be used as the hydrophilic monomer (G1). In the case of normal phase chromatography, a monomer having a primary amino group, a cyano group, or the like is converted into a hydrophilic monomer (G1). When forming affinity chromatography, a monomer having a functional group (eg, a hydroxyl group, a carboxyl group, an amino group, etc.) reactive with the ligand to be used may be used as a hydrophilic monomer (G1). The monomer having a hydrophilic hydroxyl group or the like may be used as the hydrophilic monomer (G1) when forming a gel permeation chromatography.

(Production of the packing material for liquid chromatography according to the invention of claim 5) The packing material for liquid chromatography according to the invention of claim 5 is the above-mentioned packing material.
Wherein the hydrophilic polymer fine particles (H2) having a high degree of crosslinking and having effective pores obtained by the production method according to the invention described in (1) are used as a filler for liquid chromatography. Therefore, the steps up to obtaining the hydrophilic polymer fine particles (H2) are performed in the same manner as the above-described method for producing the hydrophilic polymer fine particles (H2).

Further, as described in claim 6, by using a monomer having a functional group (J) which can be converted into an ion exchange group by a chemical reaction as the monomer (G2),
The hydrophilic polymer fine particles (H2) can be used as a filler for liquid chromatography, which is a filler for ion exchange. In this case, examples of the monomer (G2) include a monomer containing a functional group that can be converted into a cation exchange group or an anion exchange group by the above-described chemical reaction.

When used as a filler for liquid chromatography, a column for liquid chromatography can be constructed by packing the above hydrophilic polymer fine particles (H2) into a column made of stainless steel or the like. Upon filling, an appropriate method can be used, and particularly, a wet method (slurry method) is preferably used. That is, the hydrophilic polymer fine particles (H2) can be filled by dispersing a predetermined amount in a dispersion medium such as a solvent used as an eluent, and then press-fitting the column with a packer or the like.

When the hydrophilic polymer fine particles (H2) are used as a filler for liquid chromatography, by appropriately selecting the monomer (G2) and the chemical reaction method, it can be applied to various separation modes of liquid chromatography. Can be done. For example, in the case of ion-exchange chromatography, a monomer (G2) having a functional group (J) that produces an ion-exchange group by a chemical reaction may be used. A monomer that generates a hydrophobic group may be used as the monomer (G2). When normal phase chromatography is configured, a monomer that generates a primary amino group, a cyano group, or the like by a chemical reaction is used as the monomer (G2). G2) may be used. In the case of configuring affinity chromatography, a monomer which produces a functional group (eg, a hydroxyl group, a carboxyl group, an amino group, etc.) reactive with a ligand to be used by a chemical reaction is used. It may be used as a monomer (G2), and when constituting gel permeation chromatography, a monomer which produces a hydrophilic hydroxyl group by a chemical reaction may be used as a monomer. It may be used as (G2).

The substances to be measured of the packing material for liquid chromatography produced by the above invention include the following. For ion exchange chromatography,
Catecholamine derivatives such as adrenaline and noradrenaline; nucleotides such as AMP and GMP; peptides such as endorphin and insulin; proteins such as glycated hemoglobin, albumin, and myoglobin.

In the case of ion chromatography, anions such as chloride, nitrate and nitrite; cations such as sodium, ammonium and potassium. In the case of reverse phase chromatography, drugs such as antiarrhythmic agents and psychotropic drugs; organic acids such as oxalic acid and maleic acid; vitamins; proteins such as albumin and myoglobin;

In the case of normal phase chromatography, sugars such as mannose and glucose; steroids such as progesterone and testosterone; phospholipids such as phosphatidylcholine. In the case of gel permeation chromatography, synthetic polymers such as polyacrylate and polyethylene glycol; polysaccharides such as dextran; biopolymers such as various peptides, nucleic acids, and proteins. As the eluent, known ones, for example, a buffer containing an inorganic salt or an organic acid, a mixed solution of a water-soluble organic solvent and water, or the like can be used.

(Function) In the method for producing polymer fine particles according to the first aspect of the present invention, the hydrophobic crosslinkable monomer (B) is added to the hydrophobic polymer fine particles (A) dispersed in the aqueous dispersion medium. Or a monomer mixture (D) composed of a hydrophobic crosslinkable monomer (B) and a hydrophobic non-crosslinkable monomer (C), and an organic solvent (E), and a polymerization initiator (F) Polymerizes in the presence of In this case, since the degree of crosslinking is increased by the hydrophobic crosslinking monomer (B), the pressure resistance of the finally obtained polymer fine particles (H1) is enhanced.

In addition, since only the hydrophobic monomer is absorbed by the hydrophobic polymer fine particles (A) and polymerized, a polymer fine particle skeleton having almost no hydrophilic groups can be obtained.

On the other hand, while the above-mentioned hydrophobic crosslinkable monomer (B) or monomer mixture (D) is absorbed and polymerized, a hydrophilic monomer (G1) is added to continue the polymerization. Since the layer formed around the polymer fine particle skeleton contains the above-mentioned hydrophilic monomer polymer, the obtained polymer fine particles (H1) are made of a hydrophilic monomer (G1) derived from the hydrophilic monomer (G1). The hydrophilic group is hydrophilic due to the hydrophilic group, and the hydrophilic group is unevenly distributed near the surface of the hydrophilic polymer fine particles (H1). Therefore, the polymer fine particles (H
1) can be obtained, and the response speed of the hydrophilic group due to a change in the external environment can be increased, that is, the effect of shortening the equilibrium time can be obtained.

In the method for producing polymer fine particles according to the second aspect of the present invention, the hydrophobic crosslinkable monomer (B) or the hydrophobic crosslinkable monomer (B) is added to the hydrophobic polymer fine particles (A) dispersed in the aqueous dispersion medium. Crosslinkable monomer (B) and hydrophobic non-crosslinkable monomer (C)
And the organic solvent (E) is absorbed to polymerize in the presence of the polymerization initiator (F). In this case, since the degree of crosslinking is increased by the hydrophobic crosslinking monomer (B), the pressure resistance of the finally obtained polymer fine particles (H2) is enhanced.

In addition, since only the hydrophobic monomer is absorbed by the hydrophobic crosslinkable monomer (A) for polymerization, it is possible to obtain a polymer fine particle skeleton having almost no hydrophilic groups. On the other hand, it has a functional group (J) that can be converted to a hydrophilic group (I) by a chemical reaction during the process of absorbing and polymerizing the hydrophobic crosslinkable monomer (B) or monomer mixture (D). Since the polymerization is continued by adding the monomer (G2), the layer formed around the polymer fine particle skeleton is
2), the polymer microparticles (H2) obtained by the subsequent chemical reaction have the hydrophilic group (I) derived from the monomer (G2), and the hydrophilic group is hydrophilic. It is unevenly distributed near the surface of the polymer fine particles (H2). Accordingly, it is possible to obtain the polymer fine particles (H2) having excellent pressure resistance as described above while having hydrophilicity, and to increase the response speed of the hydrophilic group due to a change in the external environment, that is, to shorten the equilibration time. The effect is obtained.

According to the third aspect of the present invention, since a packing material for liquid chromatography using hydrophilic polymer fine particles (H1) having excellent pressure resistance and hydrophilicity as described above can be obtained, various liquids can be obtained. A column that can be suitably used for chromatography can be configured.

In the fourth aspect of the present invention, since the hydrophilic monomer (G1) is a monomer having an ion exchange group, the obtained hydrophilic polymer fine particles (H1) can be used as an ion exchange filler. Accordingly, the packing material for liquid chromatography suitable for ion exchange chromatography can be obtained.

According to the fifth aspect of the present invention, since a filler for liquid chromatography using hydrophilic polymer fine particles (H2) having excellent pressure resistance and hydrophilicity as described above can be obtained, various liquids can be obtained. A column that can be suitably used for chromatography can be configured.

In the invention according to claim 6, the monomer (G
Since 2) is a monomer having a functional group that generates an ion exchange group by a chemical reaction, the obtained hydrophilic polymer fine particles (H2) are used as a filler for ion exchange, and are therefore suitable for ion exchange chromatography. It can be a packing material for liquid chromatography.

[0098]

The present invention will be clarified by the following non-limiting examples.

Reference Example 1 Preparation of hydrophobic polymer fine particles A-1: 200 g of methyl methacrylate (manufactured by Wako Pure Chemical Industries, Ltd.)
Was added to 1200 g of ion-exchanged water. After heating to 70 ° C. under a nitrogen atmosphere with stirring, 10 mL of a 0.5% by weight aqueous solution of potassium persulfate (manufactured by E. Merk) and 2% by weight
10 mL of an aqueous solution of sodium styrenesulfonate (manufactured by Wako Pure Chemical Industries, Ltd.) was added dropwise to the reaction system. The polymerization was carried out at 70 ° C. for 24 hours, and the reaction product was filtered.
I got Observation with an electron microscope (manufactured by JEOL Ltd.) revealed that the particles were spherical particles with an average particle size of 0.8 μm and a CV
The value was 3.5%.

Reference Example 2 Preparation of hydrophobic polymer fine particles A-2: 5 g of Aerosol OT (manufactured by Wako Pure Chemical Industries) and 35 g of polyvinylpyrrolidone (manufactured by Wako Pure Chemical Industries), 2,2′- as a polymerization initiator 15 g of azobis- (2,4-dimethylvaleronitrile) (manufactured by Wako Pure Chemical) was dissolved in 350 mL of methanol (manufactured by Wako Pure Chemical). 50 g of ion-exchanged water and 70 of styrene (manufactured by Kishida Chemical) are added to this solution.
g under a nitrogen atmosphere with stirring.
Polymerization was performed at 10 ° C. for 10 hours. The reaction product was washed with water and methanol to obtain hydrophobic polymer fine particles A-2. Observation with an electron microscope revealed that the particles were spherical particles, the average particle size was 1.5 μm, and the CV value was 4.2%.

(Example 1) Hydrophobic polymer fine particles (A-
1) A 3% by weight aqueous solution of polyvinyl alcohol (manufactured by Nippon Synthetic Chemical) 200 ml of an aqueous solution of 0.5% by weight of sodium lauryl sulfate (manufactured by Wako Pure Chemical Industries, Ltd.)
mL was added and stirred at room temperature for 1 hour (A-1 dispersion). On the other hand, 50 g of triethylene glycol dimethacrylate (manufactured by Kishida Chemical Co.) as a hydrophobic crosslinkable monomer
And 20 mL of isoamyl alcohol, and 1.0 g of benzoyl peroxide (manufactured by Wako Pure Chemical Industries, Ltd.) as a polymerization initiator
Was dissolved. 900 mL of 0.5% by weight sodium lauryl sulfate aqueous solution was added thereto, and the mixture was homogenized (IK
A Labotechnik) at 24,000 rpm for 15 minutes. The obtained emulsion was added to the above-mentioned A-1 dispersion and stirred at room temperature for 24 hours to allow the monomer, the polymerization initiator and the organic solvent to be absorbed by the fine polymer particles A-1. Thereafter, the temperature of the reaction system was raised to 70 ° C., and polymerization was performed for 2 hours. 2
After a lapse of time, 20 g of methacrylic acid (manufactured by Wako Pure Chemical Industries, Ltd.) was added as a hydrophilic monomer, and polymerization was further performed at 70 ° C. for 2 hours. Washed with ion-exchanged water and methanol and dried,
Thus, hydrophilic polymer fine particles (H-1) were obtained.

(Example 2) Hydrophobic polymer fine particles (A-
2) 3 wt% aqueous solution of polyvinyl alcohol (manufactured by Nippon Synthetic Chemical Industry) 200 ml in 400 ml of 0.5 wt% sodium lauryl sulfate (manufactured by Wako Pure Chemical Industries) aqueous solution in which 1 g is dispersed.
mL was added and stirred for 1 hour at room temperature (A-2 dispersion). On the other hand, 50 g of divinylbenzene (manufactured by Kishida Chemical Co.) and 20 mL of toluene are used as hydrophobic crosslinkable monomers.
And 1.0 g of benzoyl peroxide (manufactured by Wako Pure Chemical Industries, Ltd.) was dissolved as a polymerization initiator. 900 mL of a 0.5% by weight aqueous solution of sodium lauryl sulfate was added thereto, and the mixture was added with a homogenizer (manufactured by IKA Labotechnik) to 2400%.
Homogenized at 0 rpm for 15 minutes. The obtained emulsion is added to the above-mentioned A-2 dispersion and stirred at room temperature for 24 hours, and the monomer, the polymerization initiator and the organic solvent are mixed with polymer fine particles A.
-2. Thereafter, the temperature of the reaction system was raised to 70 ° C.
Polymerization was carried out for hours. After 2 hours, 2- as a hydrophilic monomer
Hydroxyethyl methacrylate (Shin-Nakamura Chemical Co., Ltd.) 2
5 g was added, and polymerization was further performed at 70 ° C. for 2 hours. After washing with ion-exchanged water and methanol and drying, hydrophilic polymer fine particles (H-2) were obtained.

(Comparative Example 1) In this example, a hydrophobic crosslinkable monomer and a hydrophilic monomer were mixed, and the mixture was absorbed by hydrophobic polymer fine particles (A) to carry out polymerization. Based on the method of Example 1, a mixture of 25 g of triethylene glycol dimethacrylate, 25 g of methacrylic acid, and 20 mL of isoamyl alcohol was absorbed into 1 g of the hydrophobic crosslinkable fine particles (A-1), and the mixture was polymerized at 80 ° C. for 8 hours. Thereafter, the same treatment as in Example 1 was performed to obtain hydrophilic polymer fine particles (H-3).

(Comparative Example 2) In this example, the pore radius was 100
Particles of less than Å were prepared. The same operation as in Example 1 was carried out except that the organic solvent isoamyl alcohol in Example 1 was changed to 5 mL, to obtain hydrophilic polymer fine particles (H-4).

(Comparative Example 3) In this example, the pore radius was 100
Particles of less than Å were prepared. The same operation as in Example 2 was carried out except that the amount of the organic solvent toluene in Example 2 was changed to 5 mL,
As a result, hydrophilic polymer fine particles (H-5) were obtained.

(Comparative Example 4) In this example, the pore radius was 100
Particles exceeding 0 ° were prepared. The same operation as in Example 2 was carried out except that the amount of the organic solvent toluene in Example 2 was changed to 100 mL, to obtain hydrophilic polymer fine particles (H-6).

(Comparative Example 5) In this example, Japanese Patent Publication No. 8-719
Particles were prepared by the method described in Japanese Patent Publication No. 7 (a method in which the particle size was not controlled). 20 g of isoamyl alcohol and 1.0 g of benzoyl peroxide (manufactured by Wako Pure Chemical Industries, Ltd.) as a polymerization initiator were dissolved in 50 g of triethylene glycol dimethacrylate (manufactured by Kishida Chemical), and 4% by weight polyvinyl alcohol (manufactured by Nippon Synthetic Chemical Company) 200m aqueous solution
L was added and stirred. Thereafter, the temperature of the reaction system was raised to 80 ° C., and polymerization was performed for 2 hours. Thereafter, 30 g of methacrylic acid (manufactured by Wako Pure Chemical Industries, Ltd.) was added as a hydrophilic monomer,
Polymerization was carried out at 2 ° C. for 2 hours. After washing with ion-exchanged water and methanol and drying, hydrophilic polymer fine particles (H-7) were obtained.

(Evaluation) The above Examples 1 and 2 and Comparative Examples 1 to
About the polymer fine particles obtained in 5, a particle size distribution measurement and a pore distribution measurement, a swelling resistance test, a pressure resistance test,
Specific measurements were performed and evaluated in the following manner, respectively.

Particle Size Distribution Measurement and Pore Distribution Measurement Each hydrophilic polymer fine particle was observed with a transmission electron microscope (manufactured by JEOL Ltd.), and the particle size distribution was measured from a micrograph. The results are shown in Tables 1 and 2 below. In Examples 1 and 2 and Comparative Examples 1 to 3 (H-1 to H-5), a uniform particle size distribution was obtained. In Comparative Example 4 (H-6), some of the particles were agglomerated, and the variation in the particle size distribution was large. In Comparative Example 5 (H-7), the variation was very large. This is because H-7 is manufactured based on a method in which the particle size is not controlled.

The average pore radius was measured using a pore distribution measuring device (manufactured by Carlo Elba). Similarly, the results are shown in Table 1 below.
And Table 2. Examples 1 and 2 and Comparative Examples 1 and 5 (H-
1, 2, 3, 7), the pore radius is 100 to 1000 °.
Was in the range. Comparative Examples 2 and 3 (H-4, 5)
Less than 00 °, and more than 1000 ° in Comparative Example 4 (H-6).

[0111]

[Table 1]

[0112]

[Table 2]

Swelling resistance test The polymer fine particles H-1 (Example 1) and H-3 (Comparative Example 1) having a carboxyl group were packed in a column, and then passed through a solvent having different pH and ionic strength. The swelling resistance was confirmed by examining the pressure loss after the solution was liquefied.

1.0 g of each polymer microparticle was taken, and 50 mM
After dispersing in 30 mL of a phosphate buffer (pH = 6.0) and sonicating for 5 minutes, the mixture was thoroughly stirred. The whole amount was injected into a packer (manufactured by Umedani Seiki Co., Ltd.) to which a stainless steel empty column (6 mm diameter × 40 mm) was connected. A liquid feed pump (manufactured by Sanuki Kogyo Co., Ltd.) was connected to the packer, and filling was performed at a constant pressure of 200 kg / cm ² .

A liquid sending pump (LC-9) was applied to the obtained column.
A: Shimadzu Corporation) was connected, and a 50 mM phosphate buffer solution (pH = 5.0: A solution) was fed at a flow rate of 1.5 mL / min for 30 minutes. After that, the passing liquid is 300 mM
The solution was changed to a phosphate buffer (pH = 8.0: B solution). Fluctuation of the pressure indicated on the liquid sending pump during liquid sending was monitored. The results are shown in Table 3 below.

[0116]

[Table 3]

As is clear from Table 3, polymer fine particles H
It was found that -1 (Example 1) had less swelling / shrinkage and quicker equilibration to the external environment than the polymer fine particles H-3 (Comparative Example 1). This is presumably because the polymer microparticles H-1 have few hydrophilic groups inside the polymer microparticles, and the hydrophilic groups are unevenly distributed on the surface.

Pressure Resistance Test With respect to the hydrophilic polymer fine particles H-2 (Example 2) and H-6 (Comparative Example 4), liquids having different flow rates were passed through the packed columns, respectively. The pressure resistance was evaluated by examining the pressure loss.

Each hydrophilic polymer fine particle was subjected to the swelling resistance test described above.
Packed in a stainless steel column (filling pressure 50k)
g / cm^Two ), Connect the column to the liquid sending pump,
Was passed at 1 mL / min for 30 minutes. After that, the flow rate
0.5 mL / min was advanced by 10 minutes at each flow rate.
The pressure display of the liquid sending pump was read. As a result, the parent
In the aqueous polymer fine particles H-2, the relationship between the flow velocity and the pressure loss
Is a pressure of 400 kg / cm ^Two Also shows linearity
Was. After passing the liquid, a part of the polymer fine particles H-2 was removed from the column.
It was observed with a scanning electron microscope.
I couldn't help.

On the other hand, hydrophilic polymer fine particles H-6
Showed linearity only up to a pressure of 100 kg / cm ² . According to observation with an electron microscope, deformation and breakage of part of the polymer fine particles H-6 after the passage were observed. Thus, it was found that with H-6 having a pore radius exceeding 1000 °, the pressure resistance performance was significantly inferior.

Measurement Examples (a) Examples of Measurement of Hemoglobins by Ion Exchange Polymer fine particles having carboxyl groups H-1 (Example 1), H-3 (Comparative Example 1), H-4 (Comparative Example 2) and H
For -7 (Comparative Example 5), hemoglobins in human blood were separated. First, each of the polymer fine particles was packed into a stainless steel column having a diameter of 4.6 mm and a diameter of 35 mm in the same manner as described above, and the sample was measured under the following measurement conditions.

Measurement conditions System: Liquid pump: LC-9A (manufactured by Shimadzu Corporation) Autosampler: ASU-420 (manufactured by Sekisui Chemical Co., Ltd.) Detector: SPD-6AV (manufactured by Shimadzu Corporation) Eluent: Eluent A: 100 mM Phosphate buffer (pH 5.7) Eluent B: 300 mM phosphate buffer (pH 7.2) Flow eluent A for 0 to 1 minute from the start of measurement, and dissolve for 1 to 1.1 minutes Liquid B was flowed, and eluent A was flowed for 1.1 to 2 minutes. Flow rate: 1.5 ml / min Detection wavelength: 415 nm Sample injection volume: 10 μl

Measurement Sample Blood of a healthy human was collected with sodium fluoride, and this was hemolyzed with a hemolytic diluent (0.1% by weight of Triton X-100 in phosphate buffer (pH 7.0)) and diluted 150-fold. To obtain a measurement sample.

Measurement Results Chromatograms obtained by measuring the sample under the above measurement conditions are shown in FIGS. FIG. 1 shows the results obtained using a column of H-1 as a filler, FIG. 2 shows the results obtained using a column of H-3, FIG. 3 shows the results obtained using a column of H-4, and FIG. 4 shows the results obtained using a column of H-7. . Peak 1 is hemoglobin A1a (HbA1a) and hemoglobin A1b (HbA1b), peak 2 is hemoglobin F (HbF), and peak 3 is unstable hemoglobin A1c.
(Unstable HbA1c), peak 4 indicates stable hemoglobin A1c (stable HbA1c), and peak 5 indicates hemoglobin A0 (HbA0).

In FIG. 1 (using H-1), separation was possible in a short time, and each peak was sharp. In contrast, FIG.
In (using H-3), each peak was not sharp, and after one measurement, the inside of the column was equilibrated to the measurement start state (100% solution A) for the next sample measurement. It takes a long time. In the case of H-3, it is thought that the swelling / shrinking is likely due to the hydrophilic functional groups existing abundantly inside the particles, and it takes a long time for equilibration. FIG. 3 (H-4
Use), each peak is broadened and the separation is insufficient. This is presumably because the pore radius is small, so that the ion exchange groups on the inner surface of the pores do not act effectively and separation by sufficient ion exchange reaction is not performed. FIG. 4 (H-
7), each peak is further expanded, and the measurement time is extended. This is considered to be due to the non-uniformity of the particle size distribution of the polymer fine particles.

(B) Example of Measurement of Polymeric Substance by Gel Permeation Chromatography Polymer fine particles H-2 (Example 2) and H-5 (Comparative Example 3) having hydrophilic hydroxyl groups were polyethylene Glycol separation was performed. First, each of the polymer fine particles was packed into a stainless steel column having a diameter of 7.5 mm and a size of 150 mm in the same manner as described above, and the sample was measured under the following measurement conditions.

Measurement condition system: Liquid sending pump: LC-9A (manufactured by Shimadzu Corporation) Auto sampler: ASU-420 (manufactured by Sekisui Chemical Co., Ltd.) Detector: Shodex RI SE-51 (manufactured by Showa Denko KK) Eluent: distilled water Flow rate: 1.2 ml / min

An aqueous solution of polyethylene glycol (manufactured by Wako Pure Chemical Industries) having a molecular weight of 200 to 7500 for the measurement sample was used.

FIGS. 5 and 6 show the chromatograms obtained as a result of the measurement . FIG. 5 shows the results obtained using a column of H-2, and FIG. 6 shows the results obtained using a column of H-5. Peak 6 indicates a molecular weight of 7,500, peak 7 indicates a molecular weight of 2,000, peak 8 indicates a molecular weight of 1,000, peak 9 indicates a molecular weight of 400, and peak 10 indicates ethylene glycol. As is clear from FIG. 5, H-2 could be separated in a short time, and each peak was sharp. On the other hand, as is apparent from FIG. 6, in the case of H-5, each peak could hardly be separated. This is because the pore radius is too small,
It is considered that the separation by the sieving effect was not sufficiently performed.

(Example 3) Hydrophobic polymer fine particles (A-
1) A 3% by weight aqueous solution of polyvinyl alcohol (manufactured by Nippon Synthetic Chemical) 200 ml of an aqueous solution of 0.5% by weight of sodium lauryl sulfate (manufactured by Wako Pure Chemical Industries, Ltd.)
mL was added and stirred at room temperature for 1 hour (A-1 dispersion). On the other hand, 50 g of triethylene glycol dimethacrylate (manufactured by Kishida Chemical Co.) as a hydrophobic crosslinkable monomer
And 20 mL of isoamyl alcohol, and 1.0 g of benzoyl peroxide (manufactured by Wako Pure Chemical Industries, Ltd.) as a polymerization initiator
Was dissolved. 900 mL of 0.5% by weight sodium lauryl sulfate aqueous solution was added thereto, and the mixture was homogenized (IK
A Labotechnik) at 24,000 rpm for 15 minutes. The obtained emulsion was added to the above-mentioned A-1 dispersion and stirred at room temperature for 24 hours to allow the monomer, the polymerization initiator and the organic solvent to be absorbed by the fine polymer particles A-1. Thereafter, the temperature of the reaction system was raised to 80 ° C., and polymerization was performed for 2 hours. Thereafter, 30 g of 2-hydroxyethyl methacrylate (manufactured by Shin-Nakamura Chemical Co., Ltd.) was added as a monomer, and polymerization was further performed at 70 ° C. for 2 hours. It was washed with ion-exchanged water and methanol and dried. To 10 g of the obtained polymer microparticles, 50 mL of a 5% by weight aqueous sodium hydroxide solution was added, and 80 ° C.
For 20 hours to carry out a hydrolysis reaction, dissociate the 2-hydroxyethyl group and convert it into a carboxyl group, to obtain hydrophilic polymer fine particles (H2-1).

(Example 4) Hydrophobic polymer fine particles (A-
2) 3 wt% aqueous solution of polyvinyl alcohol (manufactured by Nippon Synthetic Chemical Industry) 200 ml in 400 ml of 0.5 wt% sodium lauryl sulfate (manufactured by Wako Pure Chemical Industries) aqueous solution in which 1 g is dispersed.
mL was added and stirred for 1 hour at room temperature (A-2 dispersion). On the other hand, 50 g of divinylbenzene (manufactured by Kishida Chemical Co.) and 20 mL of toluene are used as hydrophobic crosslinkable monomers.
And 1.0 g of benzoyl peroxide (manufactured by Wako Pure Chemical Industries, Ltd.) was dissolved as a polymerization initiator. 900 mL of a 0.5% by weight aqueous solution of sodium lauryl sulfate was added thereto, and the mixture was added with a homogenizer (manufactured by IKA Labotechnik) to 2400%.
Homogenized at 0 rpm for 15 minutes. The obtained emulsion is added to the above-mentioned A-2 dispersion and stirred at room temperature for 24 hours, and the monomer, the polymerization initiator and the organic solvent are mixed with polymer fine particles A.
-2. Thereafter, the temperature of the reaction system was raised to 70 ° C.
Polymerization was carried out for hours. After 2 hours, 25 g of glycidyl methacrylate (manufactured by NOF Corporation) was added as a hydrophilic monomer,
Further, polymerization was carried out at 70 ° C. for 2 hours. It was washed with ion-exchanged water and methanol and dried.

Xylene 3 was added to 10 g of the obtained polymer fine particles.
0 mL was added and impregnated. This is dispersed in a 4% by weight aqueous solution of polyvinyl alcohol, and 1.5 mL of sulfuric acid is further added.
Was added. The mixture was reacted at 80 ° C. for 2 hours with stirring to introduce a sulfonic acid group into the glycidyl group. The obtained particles were washed with water and acetone and dried to obtain hydrophilic polymer fine particles (H2-2).

(Comparative Example 6) In this example, a hydrophobic crosslinkable monomer and a chemically reactive monomer were mixed and absorbed by hydrophobic polymer fine particles (A) to carry out polymerization. Based on the method of Example 1, a mixture of 25 g of triethylene glycol dimethacrylate and 25 g of 2-hydroxyethyl methacrylate was absorbed into 1 g of the hydrophobic crosslinkable fine particles (A-1), and polymerized at 70 ° C. for 8 hours. A chemical reaction was carried out in the same manner as in Example 3 to obtain hydrophilic polymer fine particles (H2-3).

(Comparative Example 7) In this example, the pore radius was 100
Particles of less than Å were prepared. The same operation as in Example 3 was carried out except that the organic solvent isoamyl alcohol in Example 3 was changed to 5 mL, to obtain hydrophilic polymer fine particles (H2-4).

(Comparative Example 8) In this example, the pore radius was 100
Particles of less than Å were prepared. The same operation as in Example 4 was carried out except that the organic solvent toluene in Example 4 was changed to 5 mL, to obtain hydrophilic polymer fine particles (H2-5).

(Comparative Example 9) In this example, the pore radius was 100
Particles greater than 0 ° were prepared. The same operation as in Example 4 was carried out except that the organic solvent toluene in Example 4 was changed to 100 mL, to obtain hydrophilic polymer fine particles (H2-6).

(Comparative Example 10) In this example, the Japanese Patent Publication No. 8-71
Particles were prepared according to the method described in Japanese Patent Publication No. 97 (a method in which the particle size was not controlled). 20 mL of isoamyl alcohol and 1.0 g of benzoyl peroxide (manufactured by Wako Pure Chemical Industries, Ltd.) as a polymerization initiator were dissolved in 50 g of triethylene glycol dimethacrylate (manufactured by Kishida Chemical Co.), and 4% by weight polyvinyl alcohol (manufactured by Nippon Synthetic Chemical Company) Aqueous solution 2
00 mL was added and stirred. Thereafter, the temperature of the reaction system was raised to 80 ° C., and polymerization was performed for 2 hours. Then, 2- as a monomer
30 g of hydroxyethyl methacrylate was added, and polymerization was further performed at 80 ° C. for 2 hours. It was washed with ion-exchanged water and methanol, dried, and treated in the same manner as in Example 1 to obtain hydrophilic polymer fine particles (H2-7).

(Evaluation) Examples 3 and 4 and Comparative Examples 6 to
About the polymer fine particles obtained in 10, the particle size distribution measurement and the pore distribution measurement, the swelling resistance test, the pressure resistance test,
Specific measurements were performed and evaluated in the following manner, respectively.

Measurement of Particle Size Distribution and Measurement of Pore Distribution Each hydrophilic polymer fine particle was observed in the same manner as in the evaluation method of Example 1, and the particle size distribution was measured from a micrograph. The results are shown in Tables 4 and 5 below. Examples 3 and 4 and Comparative Example 6
-8 (H2-1 to H2-5), a uniform particle size distribution was obtained. In Comparative Example 9 (H2-6), some of the particles were aggregated. In Comparative Example 10 (H2-7), the variation was very large. This is because H2-7 is manufactured based on a method in which the particle size is not controlled.

The average pore radius was measured in the same manner as in the evaluation method of Example 1. Similarly, the results are shown in Tables 4 and 5 below. Examples 3 and 4 and Comparative Examples 6 and 10 (H2-1, 2,
In (3) and (7), the pore radius was in the range of 100 to 1000 °. Comparative Examples 7 and 8 (H2-4, 5) were 100 °
And Comparative Example 9 (H2-6) exceeded 1000 °.

[0141]

[Table 4]

[0142]

[Table 5]

Swelling Resistance Test The swelling resistance test was performed on the polymer particles H2-1 having carboxyl groups (Example 3) and H2-3 (Comparative Example 6) in the same manner as in the evaluation method of Example 1. Table 6 below
Shown in

[0144]

[Table 6]

As is clear from Table 6, the polymer fine particles H
It was found that 2-1 (Example 3) had less swelling / shrinkage and quicker equilibration to the external environment than the polymer microparticles H2-3 (Comparative Example 6). This is probably because the polymer microparticles H2-1 have few hydrophilic groups inside the polymer microparticles, and the hydrophilic groups are unevenly distributed near the surface.

Pressure Resistance Test Hydrophilic Polymer Fine Particles H2-2 (Example 4) and H2-6
(Comparative Example 9) was subjected to a pressure resistance test in the same manner as in the evaluation method of Example 2. As a result, the hydrophilic polymer fine particles H2
-2, the relationship between flow velocity and pressure loss is 400 kg / pressure.
It also showed linearity in cm ² . After the passage, a part of the polymer particles H2-2 was extracted from the column and observed with an electron microscope. No deformation or breakage was observed.

On the other hand, hydrophilic polymer fine particles H2-
In No. 6, linearity was exhibited only up to a pressure of 100 kg / cm ² . According to observation with an electron microscope, deformation and breakage of part of the polymer fine particles H2-6 after the passage were observed.
Thus, it was found that with H2-6 having a pore radius exceeding 1000 °, the pressure resistance performance was significantly inferior.

Measurement Examples (a) Examples of Measurement of Hemoglobins by Ion Exchange Fine polymer particles having carboxyl groups H2-1 (Example 3), H2-3 (Comparative Example 6), H2-4 (Comparative Example 7) and For H2-7 (Comparative Example 10), hemoglobins in human blood were separated in the same manner as in the evaluation method of Example 1.

Chromatograms obtained as a result of the measurement are shown in FIGS. FIG. 7 shows the results obtained using a column of H2-1, FIG. 8 shows the results obtained using a column of H2-3, FIG. 9 shows the results obtained using a column of H2-4, and FIG. 10 shows the results obtained using a column of H2-7. .

In FIG. 7 (using H2-1), separation was possible in a short time, and each peak was sharp. In contrast, FIG.
In (using H2-3), each peak is not sharp, and after one measurement, the inside of the column is equilibrated to the measurement start state (100% solution A) for the next sample measurement. It takes a long time. In the case of H2-3, it is thought that the swelling / shrinking is likely due to the hydrophilic functional group existing abundantly inside the particles, and it takes a long time for equilibration. FIG. 9 (H
2-4), each peak is broadened, and the separation is insufficient. This is presumably because the pore radius is small, so that the ion exchange groups on the inner surface of the pores do not act effectively and separation by sufficient ion exchange reaction is not performed. FIG.
At 0 (using H2-7), each peak is further expanded, and the measurement time is extended. This is considered to be due to the non-uniformity of the particle size distribution of the polymer fine particles.

(B) Example of Measurement of Polymeric Substance by Gel Permeation Chromatography Evaluation method of polymer fine particles H2-2 (Example 4) and H2-5 (Comparative Example 8) having hydrophilic hydroxyl groups in Example 2 In the same manner as in the above, polyethylene glycol as a polymer compound was separated.

FIGS. 11 and 12 show the chromatograms obtained as a result of the measurement . FIG.
1 is obtained using a column of H2-2 as a filler, and FIG. 12 is obtained using a column of H2-5 as a filler. As is clear from FIG. 11, H2-2 could be separated in a short time, and each peak was sharp. On the other hand, as is clear from FIG. 12, in the case of H2-5, each peak could hardly be separated. This is probably because the pore radius was too small and the separation by the sieving effect was not sufficiently performed.

[0153]

According to the method for producing polymer fine particles according to the first aspect of the present invention, the hydrophobic polymer fine particles (A) are
A monomer mixture (D) comprising a hydrophobic crosslinkable monomer (B) or a hydrophobic crosslinkable monomer (B) and a hydrophobic noncrosslinkable monomer (C), and an organic solvent (E). It is absorbed and polymerized in the presence of the polymerization initiator (F), so that the hydrophobic crosslinkable monomer (B) or the monomer mixture (D) is polymerized to form a highly crosslinked, effective pore. Particles having a radius are formed, and thus a porous polymer fine particle skeleton having excellent pressure resistance is formed.

In addition, during the polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D), a hydrophilic monomer (G1) is added to continue the polymerization to increase the hydrophilicity. Since the molecular fine particles (H1) are obtained, the hydrophilic polymer fine particles (H1)
In this example, hydrophilic groups derived from a hydrophilic monomer are unevenly distributed near the surface, and it is possible to obtain hydrophilic polymer fine particles (H1) that are unlikely to swell or shrink while maintaining hydrophilicity. I have. Therefore, according to the first aspect of the present invention, it is possible to provide hydrophilic polymer fine particles having a high degree of crosslinking, excellent pressure resistance, exhibiting hydrophilicity, and having a uniform particle diameter.

According to the method for producing polymer fine particles according to the second aspect of the present invention, the hydrophobic polymer fine particles (A) are added to the hydrophobic crosslinkable monomer (B) or the hydrophobic crosslinkable monomer. (B) and a monomer mixture (D) composed of a hydrophobic non-crosslinkable monomer (C), and an organic solvent (E) to be absorbed and polymerized in the presence of a polymerization initiator (F). A porous polymer having a high degree of crosslinking and having an effective pore radius formed by polymerizing the hydrophobic crosslinkable monomer (B) or monomer mixture (D), and thus having excellent pressure resistance A fine particle skeleton is formed.

In addition, it has a functional group (J) which can be converted into a hydrophilic group (I) by a chemical reaction during polymerization of the hydrophobic crosslinkable monomer (B) or monomer mixture (D). The monomer (G2) was added to continue the polymerization. After the polymerization, the functional group (J) was converted to the hydrophilic group (I) to obtain the hydrophilic polymer fine particles (H2). Polymer particles (H
In 2), a functional group (J) derived from the monomer (G2) is located near the surface.
It is possible to obtain hydrophilic polymer microparticles (H2) in which swelling and shrinkage are unlikely to occur while ensuring hydrophilicity, since the hydrophilic group (I) in which is converted is unevenly distributed. Therefore, according to the invention of claim 2, it is possible to provide hydrophilic polymer fine particles having a high degree of crosslinking, excellent pressure resistance, exhibiting hydrophilicity, and having a uniform particle size.

The hydrophilic polymer fine particles (H1) are
When used as a packing material for liquid chromatography as described in claim 3, the hydrophilic polymer fine particles (H1) are excellent in pressure resistance, so high-pressure packing is possible, and the speed of measurement in various types of chromatography is improved. And various samples can be measured accurately and repeatedly. Therefore, not only high pressure filling is possible,
It is possible to improve the measurement accuracy and extend the life of the filler.

By using a monomer having an ion exchange group as the hydrophilic monomer (G1), the hydrophilic polymer fine particles (H1) can be filled by ion exchange. Thus, a packing material suitable for ion-exchange chromatography can be provided.

The hydrophilic polymer fine particles (H2)
Is used as a packing material for liquid chromatography as described in claim 5, hydrophilic polymer fine particles (H2)
However, since it is excellent in pressure resistance, it can be packed under high pressure, the speed of measurement in various liquid chromatography can be increased, and various samples can be accurately and repeatedly measured. Therefore, not only high-pressure filling can be performed, but also measurement accuracy can be improved and the life of the filler can be extended.

As in the sixth aspect of the present invention, the hydrophilic polymer fine particles (H2) can be obtained by using a monomer having a functional group that generates an ion exchange group by a chemical reaction. Can be a filler for ion exchange,
Therefore, it is possible to provide a packing material suitable for ion exchange chromatography.

[Brief description of the drawings]

FIG. 1 is a diagram showing a chromatogram obtained when hemoglobins are analyzed using Example 1 (polymer fine particles H-1).

FIG. 2 is a view showing a chromatogram obtained when analyzing hemoglobins using Comparative Example 1 (polymer fine particles H-3).

FIG. 3 is a view showing a chromatogram obtained when analyzing hemoglobins using Comparative Example 2 (polymer fine particles H-4).

FIG. 4 is a view showing a chromatogram obtained when analyzing hemoglobins using Comparative Example 5 (polymer fine particles H-7).

FIG. 5 is a diagram showing a chromatogram obtained when polyethylene glycol is analyzed using Example 2 (polymer fine particles H-2).

FIG. 6 is a view showing a chromatogram obtained when polyethylene glycol is analyzed using Comparative Example 3 (polymer fine particles H-5).

FIG. 7 is a view showing a chromatogram obtained when hemoglobins are analyzed using Example 3 (polymer fine particles H2-1).

FIG. 8 is a view showing a chromatogram obtained when analyzing hemoglobins using Comparative Example 6 (polymer fine particles H2-3).

FIG. 9 is a view showing a chromatogram obtained when analyzing hemoglobins using Comparative Example 7 (polymer fine particles H2-4).

FIG. 10 is a diagram showing a chromatogram obtained when analyzing hemoglobins using Comparative Example 10 (polymer fine particles H2-7).

FIG. 11 shows the results obtained in Example 4 (polymer fine particles H2-2).
The figure which shows the chromatogram obtained when analyzing polyethylene glycol.

FIG. 12 shows Comparative Example 8 (polymer fine particles H2-5),
The figure which shows the chromatogram obtained when analyzing polyethylene glycol.

[Explanation of symbols]

 1 Peak of HbA1a and HbA1b 2 Peak of HbF 3 Peak of unstable HbA1c 4 Peak of stable HbA1c 5 Peak of HbA0 6 Peak of polyethylene glycol having a molecular weight of 7,500 7 Peak of polyethylene glycol having a molecular weight of 2,000 8 Peak of polyethylene glycol having a molecular weight of 1,000 9 Peak of polyethylene glycol having a molecular weight of 400 10 Peak of ethylene glycol

Claims (6)

[Claims] Claims: 1. A hydrophobic crosslinkable monomer (B) or a hydrophobic crosslinkable monomer (B) and a hydrophobic non-crosslinkable monomer are added to a hydrophobic polymer fine particle (A) dispersed in an aqueous dispersion medium. The monomer mixture (D) containing the monomer (C) and the organic solvent (E) are absorbed and polymerized in the presence of the polymerization initiator (F). By adding G1) and continuing the polymerization, it is crosslinked and has an average pore radius of 1
A method for producing polymer microparticles, comprising producing hydrophilic polymer microparticles (H1) having pores of 00 to 1000 °. 2. Hydrophobic cross-linkable monomer (B) or hydrophobic cross-linkable monomer (B) and hydrophobic non-cross-linkable monomer are added to hydrophobic polymer fine particles (A) dispersed in an aqueous dispersion medium. The monomer mixture (D) containing the monomer (C) and the organic solvent (E) are absorbed and polymerized in the presence of the polymerization initiator (F). A monomer (G2) having a functional group (J) that can be converted into (I) is added to continue polymerization, and after polymerization, the functional group (J) is converted into a hydrophilic group (I). A method for producing polymer fine particles, characterized by producing crosslinked hydrophilic polymer fine particles (H2) having pores having an average pore radius of 100 to 1000 °. 3. A hydrophobic crosslinkable monomer (B) or a hydrophobic crosslinkable monomer (B) and a hydrophobic non-crosslinkable monomer are added to the hydrophobic polymer fine particles (A) dispersed in an aqueous dispersion medium. The monomer mixture (D) containing the monomer (C) and the organic solvent (E) are absorbed and polymerized in the presence of the polymerization initiator (F). By adding G1) and continuing the polymerization, it is crosslinked and has an average pore radius of 1
A method for producing a packing material for liquid chromatography, comprising hydrophilic polymer fine particles (H1) having pores of 100 to 1000 °. 4. The method according to claim 3, wherein the hydrophilic monomer (G1) is a monomer having an ion exchange group, and the hydrophilic polymer fine particles (H1) are a filler for ion exchange. A method for producing a packing material for liquid chromatography. 5. Hydrophobic polymer particles (A) dispersed in an aqueous dispersion medium, a hydrophobic crosslinkable monomer (B) or a hydrophobic crosslinkable monomer (B) and a hydrophobic non-crosslinkable monomer. The monomer mixture (D) containing the monomer (C) and the organic solvent (E) are absorbed and polymerized in the presence of the polymerization initiator (F). A monomer (G2) having a functional group (J) that can be converted into (I) is added to continue polymerization, and after polymerization, the functional group (J) is converted into a hydrophilic group (I). A method for producing a filler for liquid chromatography, comprising hydrophilic polymer fine particles (H2) which are crosslinked and have pores having an average pore radius of 100 to 1000 °. 6. The packing material for liquid chromatography according to claim 5, wherein the hydrophilic group (I) is an ion exchange group, and the hydrophilic polymer fine particles (H2) are a packing material for ion exchange. Production method.