JPH11189601A - Manufacture of fine polymer particle and manufacture of filler for liquid chromatography - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an easy manufacture of a fine polymer particle which has a high crosslink density, hydrophilicity, no void and a hydrophilic group localized in the vicinity of a surface of the particle and a manufacture of a filler for liquid chromatography using such a fine polymer particle. SOLUTION: A hydrophilic fine polymer particle which has a high crosslink density and no void (G1) is manufactured by making a hydrophobic fine polymer particle dispersed in water (A) absorb a crosslinkable hydrophobic monomer (B) or a mixture (D) of a crosslinkable hydrophobic monomer (B) and a non- crosslinkable hydrophobic monomer (C), polymerizing thereof in the presence of a polymerization initiator (E) and further adding a hydrophilic monomer (F1) during polymerization for continuous polymerization.

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. The present invention relates to a method for producing hydrophilic polymer fine particles having no cavities 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. Further, in the case of polymer fine particles used as a carrier, the polymer fine particles can be reacted with a chemical substance or a biological substance by utilizing a hydrophilic functional group on the surface of the polymer fine particles.

[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, this method is limited to a method in which a low-molecular compound is separated from a mixture of a high-molecular compound and a low-molecular compound in a reversed-phase mode without performing a pretreatment operation. Further, since the polymer particles were porous, the obtained polymer fine particles did not have sufficient pressure resistance.
Furthermore, a certain kind of organic solvent is used as a porosifying agent for making the material porous, but the presence of this organic solvent tends to cause aggregation during the polymerization of the hydrophilic monomer added later on the way. . Therefore, the type of the hydrophilic monomer or the polymerization conditions have been limited. In particular, it was very difficult to use a monomer having an ion exchange group.

On the other hand, as a method for introducing an ion exchange group, after a nonionic monomer having a certain functional group (for example, a hydroxyl group) is polymerized, the functional group present in the polymer and the ion exchange group A method of introducing a compound by a chemical reaction has been proposed (JP-A-1-262468). However, it is generally known that it is difficult to quantitatively introduce a compound having an ion-exchange group by this method. (Yoshimaki, Hosoya, Kizen, Tanaka: Chromatography, 16 (1) 7-12 (1995)).

Japanese Patent Publication No. 8-7197 discloses a method for producing a filler for liquid chromatography comprising particles having a high degree of crosslinking, having no cavities, and having a hydrophilic group unevenly distributed near the surface. ing. However, in this method, since the 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 no cavity in the particle.

[0015]

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, to have no cavities, and to have a hydrophilic group. To provide a method for easily producing polymer fine particles having a uniform particle diameter, which are unevenly distributed near the particle surface, and a method for producing a packing material for liquid chromatography using such polymer fine particles. Is to do.

[0016]

According to the first aspect of the present invention, a hydrophobic polymer fine particle (A) dispersed in an aqueous dispersion medium is provided.
And the monomer mixture (D) containing the hydrophobic crosslinkable monomer (B) or the hydrophobic crosslinkable monomer (B) and the hydrophobic non-crosslinkable monomer (C) is absorbed to initiate polymerization. By polymerizing in the presence of the agent (E) and further adding the hydrophilic monomer (F1) during the polymerization and continuing the polymerization, the hydrophilic polymer fine particles which are crosslinked and have no voids ( G1).

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). Monomer mixture (D) comprising B) and a hydrophobic non-crosslinkable monomer (C)
Is added, a monomer (F2) having a functional group (I) which can be converted into a hydrophilic group (H) by a chemical reaction during the polymerization is added. To continue the polymerization, and after the polymerization, convert the functional group (I) into a hydrophilic group (H) to produce crosslinked hydrophilic polymer fine particles (G2) having no cavities. A method for producing polymer fine particles.

Further, according to the present invention, the hydrophobic cross-linkable monomer (B) or the hydrophobic cross-linkable monomer (B) is added to the hydrophobic polymer fine particles (A) dispersed in the aqueous dispersion medium. Monomer mixture (D) comprising B) and a hydrophobic non-crosslinkable monomer (C)
Is polymerized in the presence of the polymerization initiator (E) by the absorption of a polymerizable compound, and the hydrophilic monomer (F1) is added during the polymerization to continue the polymerization, whereby the polymer is crosslinked and has no void. This is a method for producing a packing material for liquid chromatography comprising hydrophilic polymer fine particles (G1).

In the third aspect of the present invention, preferably, as described in the fourth aspect, the hydrophilic monomer (F
As 1), a monomer having an ion exchange group is used, and the hydrophilic polymer fine particles (G1) 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 an aqueous dispersion medium are provided with a hydrophobic crosslinkable monomer (B) or a hydrophobic crosslinkable monomer (A). Monomer mixture (D) comprising B) and a hydrophobic non-crosslinkable monomer (C)
Is added, a monomer (F2) having a functional group (I) which can be converted into a hydrophilic group (H) by a chemical reaction during the polymerization is added. Then, after polymerization, the functional group (I) is converted into a hydrophilic group (H) to convert the functional group (I) into a hydrophilic group (H2). This is a method for producing a photographic filler.

In the invention according to claim 5, preferably, as in claim 6, the hydrophilic group (H) is an ion exchange group, and the hydrophilic polymer fine particles (G2)
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 dispersion 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 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.

(Hydrophilic Polymer Fine Particles (A)) The hydrophobic polymer fine particles (A) are not particularly limited, but may be used to obtain micron-sized polymer fine particles (G1) or (G2). 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 (G1) or (G2) may not be obtained, and if it exceeds 10 μm, the uniformity of the particle size may be impaired.

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 (G1) or (G2) may have large variations in particle diameter.

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 (F1) or (F2) 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 (J) used to obtain the hydrophobic polymer fine particles (A) include styrene and 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 acrylate, methyl methacrylate, ethyl acrylate, ethyl methacrylate, butyl acrylate, butyl methacrylate, 2-ethylhexyl acrylate, 2-ethylhexyl methacrylate, stearyl acrylate, stearyl methacrylate, Ethylene glycol diacrylate, polyethylene glycol diacrylate, 1,3-butylene glycol diacrylate, 1,6-hexane glycol diacrylate, neope Chill glycol diacrylate, polypropylene glycol diacrylate, ethylene glycol dimethacrylate, triethylene glycol dimethacrylate, polyethylene glycol dimethacrylate, 1,3-butylene glycol dimethacrylate,
1,6-hexane glycol dimethacrylate, neopentyl glycol dimethacrylate, polypropylene glycol dimethacrylate, trimethylolethane triacrylate, trimethylolethane trimethacrylate, trimethylolpropane triacrylate, trimethylolpropane trimethacrylate, tetramethylolpropane trimethacrylate, (Meth) acrylate derivatives such as tetramethylolmethane triacrylate and tetramethylolmethanetetraacrylate;
Aliphatic diene compounds such as 1,3-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 inventions of claims 1 and 3, the finally obtained polymer fine particles (G1) have hydrophilicity because most of the hydrophilic functional groups of the polymer fine particles (G1) are described later. This is because it is derived from the hydrophilic monomer (F1). In the invention according to claims 2 and 5, the finally obtained polymer fine particles (G2) have hydrophilicity because most of the hydrophilic functional groups of the polymer fine particles (G2) are described later. This is because the monomer (F2) is derived from the converted hydrophilic group (H).

(Hydrophobic Crosslinkable Monomer (B)) As the hydrophobic crosslinkable monomer, the monomer (J) used to constitute 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 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 J) 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 non-crosslinkable 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 fine particles (G1) or (G1) finally obtained can be obtained.
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) Used) 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 product polymer fine particles (G1) or (G2). 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 (G1) or (G2). In some cases, polymer fine particles (G1) or (G2) 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 (E) may be dissolved in the monomer mixture (D) and then dispersed in the dispersion medium. Alternatively, the polymerization initiator (E) may be dispersed in the dispersion medium separately from the polymerization initiator (E) 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). However, claims 1, 2, 3, 5
In the invention described in (1), when a swelling aid is used, a substance that leaves the swelling aid until after polymerization and leaves a cavity after being removed by a washing step is not used. This is because, when a swelling aid that finally leaves voids is used, the pressure resistance of the generated hydrophilic polymer fine particles (G1) or (G2) is reduced. However, there is no problem even if such a swelling aid is distilled off before polymerization.

The term "particles having cavities" as used in the present invention refers to hollow particles or porous particles generally referred to as particles having cavities on the particle surface or inside which can actually affect the properties of the particles. The hydrophilic polymer fine particles (G1) or (G2) obtained in the present invention do not have such a cavity.

(Polymerization Initiator (E)) The polymerization initiator (E) is not particularly limited, and a known water-soluble or oil-soluble radical polymerization initiator can be used. The amount of the polymerization initiator (E) 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 (E) 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 (E) 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 polymerization initiator (E) 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.

(Polymerization) A system containing a hydrophobic polymer fine particle (A), a hydrophobic crosslinkable monomer (B) or a monomer mixture (D) and a polymerization initiator (E) is added to a polymerization initiator (E). ) Is heated to a temperature at which radicals are generated, whereby the hydrophobic crosslinkable monomer (B) or the monomer mixture (D) is polymerized. The polymerization temperature varies depending on the type of the polymerization initiator (E), 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 (E), 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.

In the invention of claims 1 and 3, a hydrophilic monomer (F1) 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 In the state where 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 (E) added first remains, The monomer (F1) is added.

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 addition of the hydrophilic monomer (F1) 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 (F1) 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. When the process is almost completed, the hydrophilic group derived from the hydrophilic monomer (F1) is converted into hydrophilic polymer fine particles (G1).
Will be difficult to introduce. When the hydrophilic monomer (F1) is added, the polymerization initiator (E) may be further added.

(Hydrophilic monomer (F1)) The hydrophilic monomer (F1) is added to introduce a hydrophilic group near the surface of the finally obtained hydrophilic polymer fine particles (G1). Is done. The amount of the hydrophilic monomer (F1) 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 (F1) is less than 100 parts by weight, the hydrophilicity near the surface of the obtained hydrophilic polymer fine particles (G1) may be insufficient, and 50,000 parts by weight may be used. If it exceeds, aggregates may be generated.

Since most of the hydrophilic monomer (F1) is water-soluble, aggregation is likely to occur during the polymerization reaction. Therefore, it is preferable to stop the polymerization reaction in a state where the polymerization of the hydrophilic monomer (F1) does not sufficiently proceed (that is, before aggregation occurs).

It is not necessary to polymerize all of the hydrophilic monomer (F1), 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 (F1) added depends on the amount of the hydrophilic polymer fine particles (G
It will not match the amount of the hydrophilic monomer (F1) component contained in 1). In addition, unreacted hydrophilic monomer (F1)
Can be removed by washing after polymerization.

The presence of the organic solvent in the polymerization reaction system has an adverse effect on the polymerizability of the hydrophilic monomer (F1), increasing the probability of generating aggregates during the polymerization, and setting the conditions for the polymerization very low. It will be difficult. Therefore, from the viewpoint of the stability of the polymerization reaction, in addition to the problem of the formation of cavities, it is desirable that a generally used diluent (porous agent) is not used in the method of the present invention.

The finally obtained hydrophilic polymer fine particles (G
The degree of hydrophilicity of 1) is determined by the kind, amount and polymerization time of the hydrophilic monomer (F1) after the addition. The polymerization time of the hydrophilic monomer (F1) is preferably 0.3 to 40 hours from the start of the addition, and more preferably 0.1 to 40 hours.
5 to 20 hours. When the polymerization time of the hydrophilic monomer (F1) is less than 0.3 hours, it is difficult to reliably introduce the hydrophilic group onto the surface of the polymer fine particles. Too long, hydrophilic monomer (F
Aggregation of 1) is likely to occur.

The hydrophilic monomer (F1) is not particularly limited as long as it is a monomer having a hydrophilic group. For example, acrylic acid, methacrylic acid, itaconic acid, crotonic acid, Allyl sulfonic acid, maleic acid, fumaric acid, sodium styrene sulfonate, potassium styrene sulfonate, 3-sulfopropyl acrylate, 3-
Monomers having a cation exchange group such as sulfopropyl methacrylate and t-butylsulfonylacrylamide;
Anion exchange group-containing monomers such as dimethylaminoethyl (meth) acrylate, diethylaminoethyl (meth) acrylate, allylamine, methacrylate hydroxypropyltrimethylammonium chloride; acrylamide, methacrylamide, N-methylacrylamide, N-ethylacrylamide, N Acid amides such as -methylmethacrylamide, N-ethylmethacrylamide, N, N-dimethylacrylamide; glycidyl acrylate, glycidyl methacrylate, glycerin monoacrylate, glycerin monomethacrylate, 2-
Hydroxymethyl acrylate, 2-hydroxymethyl methacrylate, 2-hydroxyethyl acrylate,
Examples thereof include 2-hydroxyethyl methacrylate and acrylonitrile.

As to the method of adding the hydrophilic monomer (F1), during the polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D), the hydrophilic monomer (F1) is added. Can be dropped from the dropping funnel into the reaction system.

(Hydrophilic Polymer Fine Particles (G1)) The average particle diameter of the hydrophilic polymer fine particles (G1) obtained by the invention of claims 1, 3 and 4 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 hydrophilic polymer fine particles (G1)
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 (G1) having a high degree of crosslinking can be obtained.

Since the hydrophilic monomer (F1) is added and polymerized, the hydrophilic polymer fine particles (G1) have hydrophilicity. In particular, the hydrophilic monomer (F1) is added during the polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D) and is polymerized. (A), the hydrophobic crosslinkable monomer (B) and the hydrophobic non-crosslinkable monomer (C) have few hydrophilic groups. It is unevenly distributed and the number of hydrophilic groups is reduced inside the hydrophilic polymer fine particles (G1). Therefore, the polymer particles have hydrophilicity due to little swelling and shrinkage, but the hydrophilic polymer fine particles (G1) have sufficient pressure resistance. The hydrophilic polymer fine particles (G1) are non-hollow, non-porous particles having substantially no effective voids inside or on the surface of the particles, and therefore have excellent pressure resistance.

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 (F2) having a functional group (I) that can be converted to a hydrophilic group (H) 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 monomer (F2) is added in a state where the polymerization initiator (E) is in the range of about 95% and the polymerization initiator (E) added first 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 (F2). 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 (F) takes place at a stage where the polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D) has not sufficiently proceeded.
Since 2) is added, the monomer (F2) 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 monomer The polymerization of the body mixture (D) is almost complete, and the monomer (F2)
Polymerization hardly occurs. When the monomer (F2) is added, the polymerization initiator (E) may be further added.

(Monomer (F2) having a functional group (I) that can be converted into a hydrophilic group (H) by a chemical reaction) The monomer (F2) is a hydrophilic polymer fine particle finally obtained. It is added to introduce a hydrophilic group near the surface of (G2). The amount of the monomer (F2) 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 (F2) is 10
If the amount is less than 0 parts by weight, the hydrophilicity near the surface of the obtained hydrophilic polymer fine particles (G2) may be insufficient, and if it exceeds 50,000 parts by weight, aggregates may be generated.

Since the monomer (F2) is liable to cause aggregation during the polymerization reaction, the polymerization reaction may be stopped in a state where the polymerization of the monomer (F2) does not proceed sufficiently (that is, before the aggregation occurs). preferable.

It is not necessary to polymerize all of the monomers (F2), including the case where the reaction is stopped before aggregation. The reaction can be stopped when the required amount of the functional group (I) is obtained. In this case, the amount of the monomer (F2) added does not match the amount of the monomer (F2) component contained in the hydrophilic polymer fine particles (G2) obtained after the polymerization. The unreacted monomer (F2) can be removed by washing after polymerization.

The presence of an organic solvent in the polymerization reaction system has an adverse effect on the polymerizability of the monomer (F2), increases the probability of generating aggregates during the polymerization, and makes it very difficult to set conditions for the polymerization. Become. Therefore, from the viewpoint of the stability of the polymerization reaction, in addition to the problem of the formation of cavities, it is desirable that a generally used diluent (porous agent) is not used in the method of the present invention.

The polymerization time of the monomer (F2) 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 (F2) is 0.3
When the time is less than the time, the polymerization rate of the monomer (F2) is insufficient, and when the time is more than 40 hours, aggregates are easily generated.

The monomer (F2) is a monomer having a functional group (I) which can be converted into a hydrophilic group (H) by a chemical reaction. The chemical reaction referred to here is a reaction (hydrolysis reaction or the like) in which the functional group (I) is converted into a hydrophilic group (H) by decomposing in some form, or a compound having a hydrophilic group (H) is functionalized. Reaction for bonding to group (I) (condensation reaction,
Esterification reaction, acylation reaction, etc.).

Examples of the functional group (I) that can be converted to the hydrophilic group (H) 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 (F2) 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 (F2), the monomer (F2) 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 (I) by Chemical Reaction) The functional group (I) of the monomer (F2) is converted into a hydrophilic group (H) by the above chemical reaction. The hydrophilic group (H) 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 (G2) 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 (F2), the functional group (I) is a methyl ester group, and the ester group is hydrolyzed by heating under alkalinity to form a hydrophilic group (H). It is converted to a certain carboxyl group. Alternatively, when glycidyl methacrylate is used as the monomer (F2), the glycidyl group as the functional group (I) is converted into a sulfonic acid as the hydrophilic group (H) by reacting with a nitrite compound under acidic conditions. Is converted to the base. The selection of the chemical reaction for the functional group (I) of the monomer (F2) can be performed from known techniques. However, the chemical reaction needs to be performed under conditions that do not damage the polymer microparticles (G2).

(Hydrophilic Polymer Fine Particles (G2)) The average particle diameter of the hydrophilic polymer fine particles (G2) 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 hydrophilic polymer fine particles (G2)
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 (G2) having a high degree of crosslinking can be obtained.

Further, since the monomer (F2) is added and polymerized, and the functional group (I) of the monomer (F2) is converted into a hydrophilic group (H), the polymer fine particles ( G2) has hydrophilicity. In particular, since the monomer (F2) 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 finally obtained hydrophilic groups are located on the surface of the polymer fine particles (G2). And the number of hydrophilic groups inside the particles is reduced. Therefore, the polymer particles have hydrophilicity due to little swelling and shrinkage, but the hydrophilic polymer fine particles (G2) have sufficient pressure resistance. The hydrophilic polymer fine particles (G2) are non-hollow, non-porous particles having substantially no void inside or on the surface of the particles, and therefore have excellent pressure resistance.

(The hydrophilic polymer fine particles (G1) obtained by the method according to the first aspect and the hydrophilic polymer fine particles (G2) obtained by the method according to the second aspect) Application) hydrophilic polymer fine particles (G1) or (G
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 (G1) or (G
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 (G
1) or (G2) is added with various immunologically active substances (rheumatic factor, human albumin, immunoglobulin, streptolysin O, C-reactive protein, alpha-fetoprotein, etc.) and immobilized, thereby providing a carrier for immunoreagents. It is also possible to use

Further, the above-mentioned monomer (F1) including a monomer having various functional groups (ion exchange group; nonionic hydrophilic group such as hydroxyl group or cyano group; hydrophobic group such as alkyl group). And the obtained polymer fine particles (G1)
Is packed in a stainless steel column or the like to form a chromatography column. A monomer having a functional group (I) that can be converted into various functional groups (ion exchange group; nonionic hydrophilic group such as hydroxyl group and cyano group; hydrophobic group such as alkyl group) by a chemical reaction. Using (F2), the polymer fine particles (G
A column for chromatography can also be constituted by packing 2) into a column made of stainless steel or the like.

(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.
Wherein the hydrophilic polymer fine particles (G1) having a high degree of crosslinking and having no cavities 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 (G1) are as follows:
It is performed in the same manner as in the method for producing the hydrophilic polymer fine particles (G1) described above.

Further, by using a monomer having an ion exchange group as the hydrophilic monomer (F1), the hydrophilic polymer fine particles (G1) can be charged for ion exchange. And a packing material for liquid chromatography. In this case, examples of the monomer having an ion exchange group include the above-described monomer having a cation exchange group and the monomer having an anion exchange group.

When used as a filler for liquid chromatography, a column for liquid chromatography can be constituted by packing the hydrophilic polymer fine particles (G1) 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 (G1) 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 (G1) are used as a filler for liquid chromatography, the hydrophilic monomer (F1) can be appropriately selected to be applied to various separation modes of liquid chromatography. Can be. For example, in the case of ion exchange chromatography, a hydrophilic monomer (F1) having an ion exchange group may be used.
A monomer having a hydrophilic hydroxyl group or a monomer having a hydrophobic group may be used as the hydrophilic monomer (F1). In the case of constituting normal-phase chromatography, the hydrophilic monomer having a primary amino group or a cyano group may be used. In the case of configuring affinity chromatography, a monomer having a functional group (for example, a hydroxyl group, a carboxyl group, an amino group, etc.) reactive with the ligand to be used may be used. What is necessary is just to use it as a hydrophilic monomer (F1).

(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.
The hydrophilic polymer fine particles (G2) having a high degree of crosslinking and having no voids 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 (G2) are as follows:
It is performed in the same manner as in the method for producing the hydrophilic polymer fine particles (G2) described above.

Further, as described in claim 6, by using a monomer having a functional group (I) which can be converted into an ion exchange group by a chemical reaction as the monomer (F2),
The hydrophilic polymer fine particles (G2) can be used as a filler for liquid chromatography, which is a filler for ion exchange. In this case, examples of the monomer (F2) 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 constituted by packing the above hydrophilic polymer fine particles (G2) 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 (G2) 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 (G2) are used as a filler for liquid chromatography, by appropriately selecting the monomer (F2) 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 (F2) having a functional group (I) that generates an ion exchange group by a chemical reaction may be used. A monomer that produces a hydrophilic hydroxyl group or a hydrophobic group may be used as the monomer (F2). In the case of normal phase chromatography, a monomer that produces a primary amino group or a cyano group by a chemical reaction is used. In the case of affinity chromatography, a simple reaction (for example, a hydroxyl group, a carboxyl group, an amino group, or the like) that produces a functional group reactive with the ligand to be used may be used. The monomer may be used as the monomer (F2).

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. 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. Alternatively, a monomer mixture (D) composed of a hydrophobic crosslinkable monomer (B) and a hydrophobic non-crosslinkable monomer (C) is absorbed and polymerized in the presence of a polymerization initiator (E). 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 (G1) 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. In addition, since no organic solvent other than the monomer is used, there is no cavity in the polymer fine particle skeleton, which also increases the pressure resistance.

On the other hand, in the course of absorbing and polymerizing the hydrophobic crosslinkable monomer (B) or monomer mixture (D), the hydrophilic monomer (F1) is added to continue the polymerization. The polymer fine particle layer formed around the polymer fine particle skeleton,
The polymer particles (G1) thus obtained have hydrophilicity due to the hydrophilic group derived from the hydrophilic monomer (F1), and the hydrophilic group has the hydrophilic group. It is unevenly distributed near the surface of the hydrophilic polymer fine particles (G1). Therefore,
As described above, it is possible to obtain the polymer fine particles (G1) having hydrophilicity and excellent pressure resistance as described above, 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. Is 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)
Is absorbed to polymerize in the presence of the polymerization initiator (E). In this case, since the degree of crosslinking is increased by the hydrophobic crosslinkable monomer (B), the pressure resistance of the finally obtained polymer fine particles (G2) is increased.

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. In addition, since no organic solvent other than the monomer is used, there is no cavity in the polymer fine particle skeleton, which also increases the pressure resistance. On the other hand, it has a functional group (I) that can be converted into a hydrophilic group (H) by a chemical reaction during the process of absorbing and polymerizing the hydrophobic crosslinkable monomer (B) or the monomer mixture (D). Since the polymerization is continued by adding the monomer (F2), the layer formed around the skeleton of the polymer fine particles contains the monomer (F2), and is obtained by a subsequent chemical reaction. The polymer fine particles (G2) have a hydrophilic group (H) derived from the monomer (F2), and the hydrophilic groups are unevenly distributed near the surface of the hydrophilic polymer fine particles (G2). Therefore, it is possible to obtain polymer fine particles (G2) having hydrophilicity and excellent pressure resistance as described above, and
Response speed of hydrophilic group due to change of external environment is increased,
That is, the effect of shortening the equilibrium time is obtained.

According to the third aspect of the present invention, a filler for liquid chromatography using the hydrophilic polymer fine particles (G1) having excellent pressure resistance and hydrophilicity as described above can be obtained. Since has a substantially high degree of crosslinking and no cavities, it can constitute a column that can be suitably used for various liquid chromatography.

In the invention according to claim 4, since the hydrophilic monomer (F1) is a monomer having an ion-exchange group, the obtained hydrophilic polymer fine particles (G1) are filled with 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, a filler for liquid chromatography using the hydrophilic polymer fine particles (G2) having excellent pressure resistance and hydrophilicity as described above can be obtained. Since has a substantially high degree of crosslinking and no cavities, it can constitute a column that can be suitably used for various liquid chromatography.

In the invention according to claim 6, the monomer (F
Since 2) is a monomer having a functional group capable of generating an ion exchange group by a chemical reaction, the obtained hydrophilic polymer fine particles (G2) are used as a filler for ion exchange, and thus are 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 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 Shin-Nakamura Chemical Co., Ltd.) as a hydrophobic crosslinkable monomer
Then, 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 sodium lauryl sulfate solution was added thereto, and the mixture was homogenized with a homogenizer (manufactured by IKA Labotechnik) at 24000 r
Homogenized at pm for 15 minutes. The obtained emulsion was added to the above A-1 dispersion and stirred at room temperature for 24 hours to allow the monomer and the polymerization initiator 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.
After 2 hours, 30 g of methacrylic acid (manufactured by Wako Pure Chemical Industries, Ltd.) was added as a hydrophilic monomer, and polymerization was further performed at 80 ° C. for 2 hours. After washing with ion-exchanged water and methanol, drying was performed to obtain hydrophilic polymer fine particles (G-1).

(Example 2) Hydrophobic polymer fine particles (A-
2) Polymerization according to Example 1 using 1 g, 50 g of divinylbenzene (manufactured by Kishida Chemical Co., Ltd.) as a hydrophobic crosslinkable monomer; and 40 g of glycidyl methacrylate (manufactured by NOF Corporation) as a hydrophilic monomer. Was done. Post-treatment was similarly performed to obtain hydrophilic polymer fine particles (G-2).

(Example 3) Hydrophobic polymer fine particles (A-
1) Using 1 g, tetramethylolmethane triacrylate (manufactured by Shin-Nakamura Chemical Co., Ltd.) 4 as a hydrophobic crosslinkable monomer
Polymerization was carried out according to Example 1 using a mixture of 5 g and 5 g of methyl methacrylate; 25 g of diethylaminoethyl methacrylate (manufactured by NOF CORPORATION) as a hydrophilic monomer. Similarly, post-treating hydrophilic polymer fine particles (G-3)
I got

(Example 4) Hydrophobic polymer fine particles (A-
2) Using 1 g, a mixture of 45 g of divinylbenzene and 5 g of styrene was used as a hydrophobic monomer; and 20 g of sodium styrenesulfonate was used as a hydrophilic monomer to carry out polymerization according to Example 1. Post-treatment was similarly performed to obtain hydrophilic polymer fine particles (G-4).

(Comparative Example 1) In this example, a hydrophobic crosslinkable monomer and a hydrophilic monomer were mixed and absorbed by hydrophobic fine polymer 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 methacrylic acid as a hydrophilic monomer was absorbed in 1 g of the hydrophobic polymer fine particles (A-1),
Polymerization was performed at 8 ° C. for 8 hours to obtain hydrophilic polymer fine particles (G-5).

Comparative Example 2 In this example, based on the method of Example 2, a hydrophilic monomer was polymerized using a porous solvent. A liquid in which 50 g of divinylbenzene was dissolved in 100 mL of toluene (manufactured by Wako Pure Chemical Industries, Ltd.) was absorbed in 1 g of the hydrophobic polymer fine particles (A-2). After polymerization at 80 ° C. for 2 hours, 40 g of glycidyl methacrylate was added, and
For 2 hours to obtain hydrophilic polymer fine particles (G-6).

(Comparative Example 3) In this example, a monomer having an ion-exchange group was polymerized as a hydrophilic monomer in the same manner as in Example 1 except that a porous solvent was used. A solution prepared by dissolving 50 g of triethylene glycol dimethacrylate in 100 mL of isoamyl alcohol (manufactured by Wako Pure Chemical Industries, Ltd.) was absorbed into 1 g of the hydrophobic polymer fine particles (A-1), and after polymerization at 80 ° C. for 2 hours, 30 g of methacrylic acid was added. Add an additional 80
Polymerization for 2 hours at ℃, hydrophilic polymer fine particles (G-7)
I got

(Evaluation) The above Examples 1 to 4 and Comparative Examples 1 to
About 3, the state after polymerization, observation with an electron microscope, particle size distribution measurement, swelling resistance test, pressure resistance test,
Confirmation test of effect by high pressure filling, concrete measurement,
Each was performed and evaluated in the following manner.

State after polymerization: In Examples 1 to 4 and Comparative Example 1 (polymer fine particles (G-1 to G-5)), the state of polymerization was good, and no aggregation or the like occurred even in the washing step. Even after drying, uniform spherical fine powder was obtained.

On the other hand, in Comparative Example 2 (polymer fine particles G-6), some agglomerates were generated, but the amount was small and could be removed after polymerization. During the washing with methanol, the viscosity of the dispersion liquid increased and gelation was likely to occur, so the washing was interrupted halfway.

In Comparative Example 3 (polymer fine particles G-7), the viscosity sharply increased during polymerization, and gelation occurred. No decrease in viscosity was observed even after washing, and no filtration operation was possible.

Electron microscope observation: In Examples 1 to 4 and Comparative Example 1 (polymer fine particles G-1 to G-5), true spherical uniform particles were observed. On the other hand, in Comparative Example 2 (polymer fine particles G-6), abnormal large-diameter fine particles were partially observed. This seems to be due to aggregates generated during the polymerization.

Measurement of particle size distribution: The particle size distribution was measured from an electron micrograph. The results are shown in Tables 1 and 2 below. In Examples 1 to 4 and Comparative Example 1 (G-1 to G-5), numerical values indicating good monodisperse particles were obtained. In Comparative Example 2 (polymer fine particles G-6), the dispersion was large due to the presence of large particles due to the occurrence of some aggregates.
In Comparative Example 3, the particle size could not be measured due to complete aggregation.

[0114]

[Table 1]

[0115]

[Table 2]

Swelling resistance test: The polymer particles G-1 and G-5 made of methacrylic acid were examined for pressure loss in solvents having different pH and ionic strength to confirm the swelling resistance.

Take 1.0 g of each polymer fine particle, and add 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 is a stainless steel empty column (6 mm diameter x 30 mm, manufactured by Aisin Koki)
Was injected into the connected packer (Umeya Seiki). Connect a liquid feed pump (manufactured by Sanuki Kogyo) to the packer,
It was filled at a constant pressure of 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 = 6.0: A solution) was fed at a flow rate of 1.0 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.

[0119]

[Table 3]

As is clear from Table 3, polymer fine particles G
It was found that -1 (Example 1) had less swelling and shrinkage and quicker equilibration to the external environment than the polymer microparticles G-5 (Comparative Example 1). This is presumably because the polymer fine particles G-1 do not have much hydrophilic groups inside the polymer fine particles, and the hydrophilic groups are unevenly distributed on the surface.

Pressure resistance test: hydrophilic polymer fine particles G-2
With respect to (Example 2) and G-6 (Comparative Example 2), liquids having different flow rates were passed, and the pressure resistance was evaluated by examining the flow rates and the pressure loss at that time.

Each of the hydrophilic polymer fine particles was packed in a stainless steel column in the same manner as in the swelling resistance test (filling pressure 100
kg / cm ² ), the column was connected to a liquid sending pump, and the solution A was passed at 1 mL / min for 30 minutes. Thereafter, the flow rate was reduced to 0.
The pressure indication of the liquid sending pump when the liquid was passed for 10 minutes at each flow rate was read at an early rate of 5 mL / min. As a result, in the hydrophilic polymer fine particles G-2, the relationship between the flow velocity and the pressure loss showed linearity even at a pressure of 600 kg / cm ² . After the passage, a part of the polymer microparticles G-2 was withdrawn from the column,
Observation with an electron microscope revealed no deformation or breakage.

On the other hand, hydrophilic polymer fine particles G-6
Showed linearity only up to a pressure of 200 kg / cm ² . Polymer fine particles G after 200 kg / cm ² load
According to electron microscope observation, -6 was mostly deformed and partly damaged. Therefore, it can be seen that the polymer particles G-2 have a sufficient pressure resistance as compared to the polymer particles G-6.

Confirmation of Effect of High-Pressure Packing: Polymer fine particles G-2 were packed in a column at a packing pressure of 500 kg / cm ² according to Example 1. In addition, polymer fine particles G-6
At a filling pressure of 80 kg / cm ² (80
At kg / cm ² , no breakage of the filler was observed).

As the liquid sending pump, LC manufactured by Shimadzu Corporation is used.
-9A as an autosampler manufactured by Sekisui Chemical Co., Ltd.
ASU-420, SPD-, manufactured by Shimadzu Corporation as a detector
Using 6AV, flow rate 1.0mL / min, detection wavelength 254n
m, and the measurement was repeated using caffeine and theophylline as measurement samples. The change in the theoretical plate number of the caffeine peak when repeated measurements were performed under the above conditions was examined.
The results are shown in FIG. It can be seen that the polymer microparticles G-2 are superior in durability to the polymer microparticles G-6.

Measurement Examples: Polymer fine particles G-1 using methacrylic acid (Example 1), G-5 (Comparative Example 1), G-7
Regarding (Comparative Example 3), the protein was separated.
Each of the polymer microparticles was packed in a stainless steel column by the same method as in the measurement of
50 kg / cm ² , 8 for polymer fine particle G-5
The filling was performed at 0 kg / cm ² . Regarding the polymer fine particles G-7, the filling liquid did not flow regardless of the set pressure, and the polymer particles could not be packed in the column.

The apparatus used for the measurement was the same as the apparatus used for confirming the effect of high-pressure filling, and the flow rate was 0.8 mL / min, the detection wavelength was 254 nm, and the measurement samples were myoglobin (derived from horse skeleton) and α. -Chymotrypsinogen (from bovine pancreas), ribonuclease A (from bovine pancreas), and lysozyme (from chicken egg white) were used. In addition, each measurement sample is all made by Sigma.

As the eluent, 50 mL of phosphate buffer, pH = 7.0 was used as solution A, and solution A and 5 were used as solution B.
An equal mixture of 00 mM NaCl aqueous solution (pH = 7.0) was used. In addition, linear gradient elution from 100% of solution A to 100% of solution B was performed.

The obtained chromatograms are shown in FIG. 2 (results for polymer fine particles G-1) and FIG.
-5). As is clear from FIG. 2, the polymer fine particles (G-1) can be separated in a short time,
Each peak was also sharp. On the other hand, in the case of the polymer fine particles (G-5), each peak was not sharp, and after one measurement was completed, the inside of the column was brought into a measurement start state (100% solution A) for the next sample measurement. It takes a long time to equilibrate. This is considered to be because the polymer particles (G-5) cannot be packed under high pressure and the separation ability is reduced, and the polymer particles (G-5) easily swell and shrink, so that it takes time to equilibrate.

(Example 5) 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
Then, 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 sodium lauryl sulfate solution was added thereto, and the mixture was homogenized with a homogenizer (manufactured by IKA Labotechnik) at 24000 r
Homogenized at pm for 15 minutes. The obtained emulsion was added to the above A-1 dispersion and stirred at room temperature for 24 hours to allow the monomer and the polymerization initiator 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 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 to a carboxyl group to obtain hydrophilic polymer fine particles (G2-1).

(Example 6) 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) as a hydrophobic crosslinkable monomer and 1.0 g of benzoyl peroxide (manufactured by Wako Pure Chemical Industries) as a polymerization initiator were dissolved. 900 mL of a 0.5% by weight aqueous solution of sodium lauryl sulfate was added thereto, and a homogenizer (IKA Labotechni) was added.
(manufactured by k Company) at 24,000 rpm for 15 minutes. The obtained emulsion was added to the above dispersion A-2 and stirred at room temperature for 24 hours to allow the monomer and the polymerization initiator to be absorbed by the fine polymer particles A-2. Thereafter, the reaction system was heated to 70 ° C.
And polymerized for 2 hours. After 2 hours, glycidyl methacrylate (manufactured by NOF CORPORATION) 25 as a hydrophilic monomer
g was added, and polymerization was further performed at 70 ° C. for 2 hours. It was washed with ion-exchanged water and methanol and dried. 30 mL of xylene was added to 10 g of the obtained polymer microparticles for impregnation. This was dispersed in a 4% by weight aqueous solution of polyvinyl alcohol, and 1.5 mL of sulfuric acid was further 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 (G2-2).

(Example 7) 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
Then, 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 sodium lauryl sulfate solution was added thereto, and the mixture was homogenized with a homogenizer (manufactured by IKA Labotechnik) at 24000 r
Homogenized at pm for 15 minutes. The obtained emulsion was added to the above A-1 dispersion and stirred at room temperature for 24 hours to allow the monomer and the polymerization initiator 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 glycidyl methacrylate (manufactured by NOF Corporation) 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. 10 mL of a 500 mM hydrochloric acid aqueous solution was added to 10 g of the obtained polymer fine particles, and 50 mL of a 20% by weight aqueous sodium sulfite solution was further added. The mixture was allowed to react at 80 ° C. for 20 hours to introduce a sulfonic acid group into a glycidyl group, thereby obtaining hydrophilicity. Polymer fine particles (G2-3) were obtained.

(Comparative Example 4) In this example, a hydrophobic crosslinkable monomer and a chemically reactive monomer (F2) were mixed, and the mixture was absorbed by hydrophobic fine polymer particles (A) to carry out polymerization. . Based on the method of Example 5, 1 g of the hydrophobic crosslinkable fine particles (A-1)
Triethylene glycol dimethacrylate 25 g and 2
A mixture of 25 g of -hydroxyethyl methacrylate was absorbed and polymerized at 80 ° C for 8 hours. After polymerization, a chemical reaction was carried out in the same manner as in Example 5 to obtain hydrophilic polymer fine particles (G2-4).

Comparative Example 5 In this example, porous particles were prepared. 0.5% by weight sodium lauryl sulfate in which 1 g of hydrophobic polymer fine particles (A-2) are dispersed (manufactured by Wako Pure Chemical Industries, Ltd.)
200 mL of a 3% by weight aqueous solution of polyvinyl alcohol (manufactured by Nippon Synthetic Chemical Industry) was added to 400 mL of the aqueous solution, and the mixture was stirred at room temperature for 1 hour (A-2 dispersion). On the other hand, divinylbenzene (manufactured by Kishida Chemical Co.) as a hydrophobic crosslinkable monomer
50 g and 100 mL of toluene as a porosifying agent were mixed, 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 sodium lauryl sulfate solution was added thereto, and the mixture was homogenized with a homogenizer (manufactured by IKA Labotechnik) at 24,000 rpm.
m for 15 minutes. The obtained emulsion was added to the above dispersion A-2 and stirred at room temperature for 24 hours to allow the monomer and the polymerization initiator to be absorbed by the fine polymer particles A-2.
Thereafter, the temperature of the reaction system was raised to 70 ° C., and polymerization was performed for 2 hours.
Two hours later, 25 g of glycidyl methacrylate (manufactured by NOF Corporation) was added as a hydrophilic monomer, and polymerization was further performed at 70 ° C. for 2 hours. It was washed with ion-exchanged water and methanol and dried. 30 mL of xylene was added to 10 g of the obtained polymer microparticles for impregnation. This was dispersed in a 4% by weight aqueous solution of polyvinyl alcohol, and 1.5 m
L was added. Reaction at 80 ° C. for 2 hours with stirring,
A sulfonic acid group was introduced into the glycidyl group. The obtained particles were washed with water and acetone and dried to obtain hydrophilic polymer fine particles (G2-5).

(Comparative Example 6) 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). Dissolve 1.0 g of benzoyl peroxide (manufactured by Wako Pure Chemical Industries) as a polymerization initiator in 50 g of triethylene glycol dimethacrylate (manufactured by Kishida Chemical), and add 200 mL of a 4% by weight aqueous solution of polyvinyl alcohol (manufactured by Nippon Synthetic Chemical) 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 2-hydroxyethyl methacrylate was added as a hydrophilic monomer, and polymerization was further performed at 80 ° C. for 2 hours.
The polymer was washed with ion-exchanged water and methanol, dried, treated in the same manner as in Example 5, and treated with hydrophilic polymer fine particles (G2-
6) was obtained.

(Evaluation) The above Examples 5 to 7 and Comparative Examples 4 to
Particle size distribution measurement, swelling resistance test, pressure resistance test, and specific measurement of the hydrophilic polymer fine particles obtained in 6 were performed and evaluated in the following manner, respectively.

Particle size 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 an electron micrograph. The results are shown in Tables 4 and 5 below. In Examples 5 to 7 and Comparative Example 4 (polymer fine particles G2-1 to G2-4), a uniform particle size distribution was obtained. In Comparative Example 5 (polymer fine particles G2-5), some of the particles were aggregated. Comparative Example 6 (polymer fine particles G
In 2-6), the variation was very large. This is G
This is because 2-6 is manufactured based on a method in which the particle size is not controlled.

[0138]

[Table 4]

[0139]

[Table 5]

Swelling resistance test: The polymer fine particles G2-1 (Example 5) and G2-4 (Comparative Example 4) having a carboxyl group were packed in a column, and then solvents having different pH and ionic strength were used. The swelling resistance was confirmed by examining the pressure loss by passing the solution through. Each polymer fine particle
0 g was collected and 50 mM phosphate buffer (pH 6.0) 3
After dispersing in 0 mL and sonicating for 5 minutes, the mixture was stirred well. The entire volume is a stainless steel empty column (6 mm diameter x 30 m
m) was injected into the connected packer (manufactured by Umetani Seiki Co., Ltd.).
A liquid feed pump (manufactured by Sanuki Kogyo KK) 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 6 below.

[0142]

[Table 6]

As is clear from Table 6, polymer fine particles G
It was found that 2-1 (Example 5) had less swelling / shrinkage and quicker equilibration to the external environment than the polymer fine particles G2-4 (Comparative Example 4). It is considered that this is because the polymer microparticles G2-1 have few hydrophilic groups inside the polymer microparticles, and the hydrophilic groups are unevenly distributed on the surface.

Pressure test: hydrophilic polymer fine particles G2-
With respect to Sample No. 2 (Example 6) and G2-5 (Comparative Example 5), liquids having different flow rates were passed through columns packed with the respective liquids, and the pressure resistance was evaluated by examining the flow rates and the pressure loss at that time. .

Each hydrophilic polymer fine particle was packed in a stainless steel column in the same manner as in the above-mentioned swelling resistance test (filling pressure 80 k
g / cm ² ), the column was connected to a liquid sending pump, and the above solution A was passed at 1 mL / min for 30 minutes. Thereafter, the flow rate was increased by 0.5 mL / min, and the pressure display of the liquid sending pump when the liquid was passed at each flow rate for 10 minutes was read. As a result, in the hydrophilic polymer fine particles G2-2, the relationship between the flow velocity and the pressure loss showed linearity even at a pressure of 600 kg / cm ² . G2-5 showed linearity only up to a pressure of 100 kg / cm ² . After the passage, the polymer fine particles were extracted from the column and observed with an electron microscope. As a result, no deformation or damage was observed in G2-2, but some deformation or damage was observed in G2-5. This is considered to be due to a decrease in pressure resistance performance due to porosity.

Measurement Examples: Fine polymer particles G2-1 having carboxyl groups (Example 5), G2-4 (Comparative Example 4) and G2-6 (Comparative Example 6) and fine polymer particles G2- having sulfonic acid groups Regarding Example 3 (Example 7), hemoglobins in human blood were separated. First, each of the polymer fine particles was packed in a stainless steel column having a diameter of 4.6 mm and 35 mm by the same method 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. If the eluent A and eluent B cannot be separated satisfactorily even when they are passed for the above-mentioned flow time, set the flow time so that the separation can be as good as possible within the practical measurement time. changed. 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. 4 shows that the filler is G2-1,
FIG. 5 shows the results obtained using a column of G2-3, FIG. 6 shows the results obtained using a column of G2-4, and FIG. 7 shows the results obtained using a column of G2-6. Peak 5 is hemoglobin A1a (HbA1a) and hemoglobin A1b (HbA1b), peak 6 is hemoglobin F (HbF), peak 7 is unstable hemoglobin A1c (unstable HbA1c), peak 8 is stable hemoglobin A1c (stable HbA1c), Peak 9 indicates hemoglobin A0 (HbA0).

FIGS. 4 and 5 (using G2-1 and G2-3)
Could be separated in a short time, and each peak was sharp.
On the other hand, in FIG. 6 (using G2-4), each peak is not sharp, and after the completion of one measurement, the inside of the column is started to be measured for the next sample measurement (100% solution A). It takes a long time to equilibrate. In G2-4,
This is probably due to the fact that the hydrophilic functional groups abundantly present inside the particles swell and shrink easily, and it takes time to equilibrate. In FIG. 7 (using G2-6), each peak is further expanded, and the measurement time is also extended. This is considered to be due to the non-uniformity of the particle size distribution of the polymer fine particles.

[0151]

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
Absorb the hydrophobic crosslinkable monomer (B) or the monomer mixture (D) composed of the hydrophobic crosslinkable monomer (B) and the hydrophobic non-crosslinkable monomer (C), and form a polymerization initiator ( A layer having a high degree of crosslinking formed by polymerizing the hydrophobic crosslinkable monomer (B) or the monomer mixture (D) around the hydrophobic polymer fine particles (A) because of polymerization in the presence of E). Is formed, and thus a polymer fine particle skeleton excellent in pressure resistance is formed. In this case, since no organic solvent is used, no pores or cavities are formed in the polymer fine particle skeleton, whereby the pressure resistance of the polymer fine particle skeleton is also enhanced.

In addition, during the polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D), the hydrophilic monomer (F1) is added to continue the polymerization. Since the hydrophilic polymer microparticles (G1) are obtained, the hydrophilic polymer microparticles (G1) have unevenly distributed hydrophilic groups derived from the hydrophilic monomer near the surface, and while ensuring hydrophilicity, It is possible to obtain hydrophilic polymer microparticles (G1) that are unlikely to cause swelling or shrinkage. Therefore, according to the invention described in claim 1, the degree of crosslinking is high, the pressure resistance is excellent, the hydrophilicity is exhibited,
In addition, hydrophilic polymer fine particles having a uniform particle size can be provided.

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 the monomer mixture (D) composed of the hydrophobic non-crosslinkable monomer (C) are absorbed and polymerized in the presence of the polymerization initiator (E). A layer having a high degree of cross-linking formed by polymerizing the hydrophobic cross-linkable monomer (B) or the monomer mixture (D) is formed around the polymer layer, and thus a polymer fine particle skeleton excellent in pressure resistance is formed. . In this case, since no organic solvent is used, no pores or cavities are formed in the polymer fine particle skeleton, whereby the pressure resistance of the polymer fine particle skeleton is also enhanced.

In addition, during polymerization of the hydrophobic crosslinkable monomer (B) or the monomer mixture (D), a monomer having a functional group (I) which can be converted into a hydrophilic group by a chemical reaction. Since the polymerization is continued by adding the polymer (F2), the layer formed around the polymer fine particle skeleton contains the above-mentioned monomer (F2), and thus the high molecular weight obtained by the subsequent chemical reaction is obtained. The molecular fine particles (G2) have a hydrophilic group (H) converted from the monomer (F2), and the hydrophilic groups are unevenly distributed near the surface of the hydrophilic polymer fine particles (G2). It is possible to obtain hydrophilic polymer fine particles (G2) in which swelling, shrinkage, and the like are unlikely to occur while ensuring the properties. 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.

When the hydrophilic polymer fine particles are used as a filler for liquid chromatography as described in claim 3, the hydrophilic polymer fine particles (G1) have excellent pressure resistance. It can be packed under high pressure, can increase the speed of measurement in various liquid chromatography, and can measure various samples 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 (F1), the hydrophilic polymer fine particles (G1) can be charged for ion exchange. Thus, a packing material suitable for ion-exchange chromatography can be provided.

When the hydrophilic polymer fine particles (G2) are used as a filler for liquid chromatography as described in claim 5, the hydrophilic polymer fine particles (G2) have excellent pressure resistance. It can be packed under high pressure, can increase the speed of measurement in various liquid chromatography, and can measure various samples 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.

As in the sixth aspect of the present invention, the hydrophilic polymer fine particles (G2) can be obtained by using a monomer having a functional group capable of generating an ion exchange group by a chemical reaction as the monomer (F2). 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 shows Example 2 (polymer fine particles G-2) and Comparative Example 2
The figure which showed the change of the theoretical plate number at the time of performing analysis using (polymer fine particle G-6) as an initial value 100.

FIG. 2 is a view showing a chromatogram obtained when a protein is analyzed using Example 1 (polymer fine particles G-1).

FIG. 3 is a view showing a chromatogram obtained when a protein is analyzed using Comparative Example 1 (polymer fine particles G-5).

FIG. 4 is a view showing a chromatogram obtained when analyzing hemoglobins using Example 5 (polymer fine particles G2-1).

FIG. 5 is a view showing a chromatogram obtained when analyzing hemoglobins using Example 7 (polymer fine particles G2-3).

FIG. 6 is a view showing a chromatogram obtained when analyzing hemoglobins using Comparative Example 4 (polymer fine particles G2-4).

FIG. 7 is a view showing a chromatogram obtained when analyzing hemoglobins using Comparative Example 6 (polymer fine particles G2-6).

[Explanation of symbols]

 1 ... peak of myoglobin 2 ... peak of α-chymotrypsinogen 3 ... peak of ribonuclease A 4 ... peak of lysozyme 5 ... peak of HbA1a and HbA1b 6 ... peak of HbF 7 ... peak of unstable HbA1c 8 ... peak of stable HbA1c 9: HbA0 peak

 ────────────────────────────────────────────────── ─── Continued on the front page (72) Inventor Yuji Setoguchi 4560 Kaiseicho, Shinnanyo-shi, Yamaguchi Prefecture Sekisui Chemical Co., Ltd.

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. Polymerisation initiator (E) by absorbing monomer mixture (D) containing monomer (C)
, And by continuing the polymerization by adding the hydrophilic monomer (F1) during the polymerization, the crosslinked hydrophilic polymer fine particles (G
A method for producing polymer fine particles, which comprises producing 1). 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. Polymerisation initiator (E) by absorbing monomer mixture (D) containing monomer (C)
, And further added with a monomer (F2) having a functional group (I) that can be converted into a hydrophilic group (H) by a chemical reaction during the polymerization, and the polymerization is continued. By converting the functional group (I) into a hydrophilic group (H), the hydrophilic polymer fine particles (G
2. A method for producing polymer fine particles, which comprises the step of 2). 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. Polymerisation initiator (E) by absorbing monomer mixture (D) containing monomer (C)
, And by continuing the polymerization by adding the hydrophilic monomer (F1) during the polymerization, the crosslinked hydrophilic polymer fine particles (G
1) A method for producing a packing material for liquid chromatography comprising: 4. The method according to claim 3, wherein the hydrophilic monomer (F1) is a monomer having an ion exchange group, and the hydrophilic polymer fine particles (G1) 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. Polymerisation initiator (E) by absorbing monomer mixture (D) containing monomer (C)
, And further added with a monomer (F2) having a functional group (I) that can be converted into a hydrophilic group (H) by a chemical reaction during the polymerization, and the polymerization is continued. By converting the functional group (I) into a hydrophilic group (H), the hydrophilic polymer fine particles (G
2) A method for producing a packing material for liquid chromatography comprising: 6. The packing material for liquid chromatography according to claim 5, wherein the hydrophilic group (H) is an ion exchange group, and the hydrophilic polymer fine particles (G2) are a packing material for ion exchange. Production method.