JP2006137944A - Crosslinked (meth)acrylamide particles, process for their production and their use - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To solve the problem that acrylamide particles are mechanically weak, in particular, are difficult to obtain as fine particles of about 10 μm in size so as to be used in liquid chromatography, and are difficult to produce monodisperse crosslinked acrylamide particles having an arbitrary degree of crosslinking. <P>SOLUTION: The method of producing crosslinked (meth)acrylamide particles as particle size monodisperse particles comprises obtaining crosslinked (meth)acrylamide particles having an arbitrary degree of crosslinking and mechanically strong, and which can be introduced with various functional groups. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to crosslinked (meth) acrylamide particles.

  Polymerized particles containing a (meth) acrylamide monomer as a component (hereinafter referred to as (meth) acrylamide particles) have high hydrophilicity, excellent moisture retention, and low biotoxicity, making them suitable for ink binders, surface treatment agents, cosmetics, etc. Widely used. In particular, the substrate for liquid chromatography has excellent characteristics as a carrier for purification of proteins, nucleic acids, and the like because the hydrophobic interaction between the sample and the substrate is small when an aqueous eluent is used.

  Conventionally, as hydrophilic particles, particles obtained by crosslinking polysaccharides such as cellulose, dextran, or agarose are widely known. When preparing polysaccharide-based crosslinked particles, an aqueous solution of a polysaccharide and a crosslinking agent is suspended in an organic solvent in reverse phase, and the polysaccharide is crosslinked with an epoxy compound or dialdehyde as a crosslinking agent. .

  Further, (meth) acrylamide particles are prepared by polymerizing water-soluble (meth) acrylamide, and generally, a method of polymerizing by reverse phase suspension is employed (Patent Document 1, Patent Document 2, Patent Document). 3, Patent Document 4). As a production method other than the reverse phase suspension polymerization, a dispersion polymerization method in an organic solvent is known as a method for producing monodisperse particles having a uniform particle diameter of (meth) acrylamide particles (Patent Document 5). As other production methods, a polymerization method in critical carbon dioxide (Patent Document 6) and a spray method (Patent Document 7) are known.

  Further, N-alkoxymethyl (meth) acrylamide is a self-crosslinking monomer, and it is known that an alkoxy group can be eliminated by an acid catalyst and can easily bond to a self-crosslink or an amide group, an amino group or the like. For this reason, it is used for fiber processing (Patent Literature 8, Patent Literature 9), and used as a colloidal dispersant (Patent Literature 10, Patent Literature 11) or a coating film forming material (Patent Literature 12, Patent Literature 13). ing.

U.S. Pat. No. 4,070,348 (Claim 6) US Pat. No. 4,190,713 (abstract) U.S. Pat. No. 4,511,694 (Claim 1) JP 59-232101 (Claim 10) U.S. Pat. No. 4,988,568 (summary, claim 3) U.S. Pat. No. 4,748,220 (Abstract, Claim 4) JP 2003-211003 A (Claim 1) US Pat. No. 5,219,969 (abstract) US Pat. No. 5,149,943 (Background of the Invention) US Pat. No. 5,385,971 (Details of the Invention) US Pat. No. 6,316,568 (Claim 3) Japanese Patent Laid-Open No. 51-8343 (Section 3, Example 5) US Pat. No. 4,107,156 (Example 15)

  (Meth) acrylamide particles are used in various fields because of their properties as hydrophilic particles. However, there are problems in that the method for introducing a functional group is limited and the mechanical strength is weak. Further, since the mechanical strength is weak, it is difficult to make fine particles, and there remains a problem that particles having a size of 50 μm or less, particularly about 10 μm, which are used in liquid chromatography cannot be obtained.

  Another problem is that it is difficult to control the particle diameter in the method for producing (meth) acrylamide particles. This is because it is difficult to control the particle size in the reverse phase suspension polymerization method, which is a general method for producing (meth) acrylamide particles, and the seed is a method for producing monodisperse particles in the normal phase suspension polymerization method. This is because the polymerization method cannot be performed with (meth) acrylamide particles. The dispersion polymerization method can produce (meth) acrylamide particles having a uniform particle diameter, but the range of particle diameters that can be produced is as narrow as 10 μm or less, and the ratio between the crosslinkable monomer and (meth) acrylamide. However, since the particle size also changes when the particle size is changed and the particle size decreases when the degree of crosslinking is increased, the degree of crosslinking cannot be arbitrarily changed.

  An object of the present invention is to provide crosslinked (meth) acrylamide particles into which various substituents are introduced, and has excellent properties as a packing material for liquid chromatography having both high hydrophilicity and mechanical strength. The object is to provide crosslinked (meth) acrylamide particles. In addition, the production method of introducing substituents to polymer particles produced by normal phase suspension polymerization of the present invention facilitates crosslinking (meth) acrylamide particles having any degree of crosslinking and having various functional groups introduced. It is to provide a manufacturing method that can be manufactured. Another object of the present invention is to provide a method for producing crosslinked (meth) acrylamide particles having a particle size monodisperse having an arbitrary degree of crosslinking which cannot be produced by other polymerization methods by using a seed polymerization method.

The present invention relates to a unit derived from an N-alkoxymethyl (meth) acrylamide monomer represented by Chemical Formula 1 (X in Chemical Formula 1 is hydrogen or a methyl group, and R1 represents a C4 or higher alkyl group). ,
In polymer particles (hereinafter referred to as the present polymer particles) having a particle diameter of 1 to 1000 μm, including units derived from polyfunctional unsaturated monomers that can be copolymerized with the monomer represented by Chemical Formula 1,
The unit derived from the N-alkoxymethyl (meth) acrylamide monomer represented by Chemical Formula 1 is represented by Chemical Formula 2 (wherein R2 in Chemical Formula 2 is H, an alkyl group of C3 or less, or C3 or less and one or more hydroxyl groups) In the chemical formula 2, R3 is a linear, branched, or alkyl group of H, NH2, C1 to C18, a phenyl group, C24 or less and substituted with one or more hydroxyl groups. It represents either a cyclic alkyl group, —CH 2 —CO—NH 2, or R 3 is a structural unit derived from another N-alkoxymethyl (meth) acrylamide monomer, It represents bonding to a nitrogen atom derived from an N-alkoxymethyl (meth) acrylamide monomer. A in Chemical Formula 2 represents a polymer particle) and / or Chemical Formula 3 (only R4 and R5 in Formula 3 are H, an alkyl group of C6 or less, a phenyl group, a naphthyl group, an alkyl group of C6 or less and substituted with one or more hydroxyl groups, -CH2COOH, -CH2SO3H, -CH2CH2SO3H,-( CH2) n-PO- (OH) 2 (wherein n represents an integer of 1 to 4), -CHR6-PO- (OH) 2 (wherein R6 represents a linear alkyl group having 1 to 3 carbon atoms). ), -CHR7-COOH (R7 represents an amino acid side chain), oligopeptide residue, cytidine residue, guanidine residue, melamine residue, benzoguanamine residue, polyethyleneimine, polyallylamine, or , R4 and R5 are structural units derived from other N-alkoxymethyl (meth) acrylamide monomers, and other N-alkoxy via a methylene group It represents a bond to a nitrogen atom derived from a chill (meth) acrylamide monomer.A in Chemical Formula 3 represents a polymer particle.) ) Providing acrylamide particles.

架橋（メタ）アクリルアミド粒子における置換基は、化学式４や化学式５に示されるような反応によって、酸触媒によるアルコキシ基脱離反応によるアミド基やアミノ基の導入が可能であり、本重合体粒子に種々の官能基を容易に誘導することができる。

Substituents in the crosslinked (meth) acrylamide particles can be introduced with amide groups or amino groups by an alkoxy group elimination reaction with an acid catalyst by a reaction shown in Chemical Formula 4 or Chemical Formula 5, Various functional groups can be easily derived.

化学式４中のＲ１、Ｒ２及びＲ３は化学式１及び化学式２に記載の置換基と同一。化学式４中のＡは重合体粒子を表す。

R1, R2 and R3 in Chemical Formula 4 are the same as the substituents described in Chemical Formula 1 and Chemical Formula 2. A in Chemical Formula 4 represents polymer particles.

化学式５中のＲ１、Ｒ４及びＲ５は化学式１及び化学式３に記載の置換基と同一。化学式５中のＡは重合体粒子を表す。

R1, R4 and R5 in Chemical Formula 5 are the same as the substituents described in Chemical Formula 1 and Chemical Formula 3. A in Chemical Formula 5 represents polymer particles.

  When a compound having an amide group is introduced, particles into which a highly hydrophilic amide compound such as formamide, lactamide or urea is introduced become porous highly hydrophilic particles. At this time, particles obtained using a polyfunctional amide compound such as urea become particles having both high hydrophilicity and unprecedented mechanical strength, and the particle diameter can be reduced. Useful. In addition, particles into which a compound having an amide group and an ion exchange group is introduced become an ion exchanger containing a (meth) acrylamide monomer as a component. Furthermore, it is also possible to introduce polyacrylamide as a compound having an amide group and various copolymers containing acrylamide as a component.

  Moreover, it is possible for this polymer particle to introduce | transduce the compound which has an amino group which has N-H instead of an amide group. By using a compound having another ion exchange group at the same time as the compound having an amino group, for example, an ion exchanger having a betaine structure is obtained. For example, when the compound having an amino group having N—H is a compound having a carboxylic acid group, a sulfonic acid group or a phosphoric acid group at the same time, the amino group is an anion exchange group by bonding to the polymer particles. It becomes an ion exchanger having a betaine structure having an amino group and a cation exchange group at the same time.

  The polymer particles can be prepared by a normal phase suspension polymerization method, and can be prepared by a seed polymerization method using a seed particle polymer having a uniform particle diameter. Particles having an arbitrary degree of cross-linking with a polyfunctional unsaturated monomer and having a uniform particle diameter can be easily prepared, and the above-mentioned various monodispersed (meth) acrylamide particles can be produced.

  The present invention is described in detail below.

  A unit derived from an N-alkoxymethyl (meth) acrylamide monomer represented by Chemical Formula 1 and a unit derived from a polyfunctional unsaturated monomer that can be copolymerized with the monomer represented by Chemical Formula 1 Since the N-alkoxymethyl (meth) acrylamide monomer of polymer particles having a particle diameter of 1-1000 μm (hereinafter referred to as the present polymer particles) is preferably oil-soluble, the alkoxy group of Formula 1 is Since it is necessary to be a hydrophobic group, R1 in the chemical formula 1 is preferably a linear alkyl group of C4 or more. Especially when the present polymer particles are produced by a seed polymerization method, the swelling property is also improved. In consideration, R1 in Chemical Formula 1 is preferably a linear alkyl group in the range of C4 to C8.

  The unit derived from a polyfunctional unsaturated monomer in the present invention is a unit derived from an oil-soluble monomer and needs to be copolymerizable with an N-alkoxymethyl (meth) acrylamide monomer. . Examples of these are (meth) acrylic acid esters such as ethylene glycol di (meth) acrylate, diethylene glycol di (meth) acrylate, polyethylene glycol di (meth) acrylate, glycerin di (meth) acrylate, glycerin tri (meth) acrylate, Methylene bismaleimide, N, N ′-(4-methyl-1,3-phenylene) bismaleimide, N, N ′-(phenylene) bismaleimide, N, N ′-(sulfonyldi-m-phenylene) bismaleimide, etc. And bismaleimides, divinylbenzene, triallyl isocyanurate, and methylene bisacrylamide. The amount of N-alkoxymethyl (meth) acrylamide and polyfunctional unsaturated monomer can be used within the range of 99.99: 0.01 to 50:50. Considering from the viewpoint of reducing the hydrophobic effect of the monomer and the viewpoint of the strength of the obtained particles, a range of 99.95: 0.05 to 98: 2 is preferable.

  Other monomers may be copolymerized with the N-alkoxymethyl (meth) acrylamide monomer represented by Chemical Formula 1 and the polyfunctional unsaturated monomer. However, since other monomers need to be oil-soluble hydrophobic monomers, when making particles that make use of the hydrophilicity of (meth) acrylamide contained in the components using this polymer particle as a starting material, It is not preferable to add other monomers.

  The particle diameter of the polymer particles is arbitrary depending on the use, but is preferably 1 to 1000 μm, particularly preferably 5 to 200 μm in the use as a filler for liquid chromatography.

  This polymer particle can easily introduce a compound having an amide group in which at least one bond on a nitrogen atom is an N—H bond in an organic solvent under an acid catalyst. The alkoxy group in the N-alkoxymethylamide group is eliminated, and the amide group nitrogen atom in the polymer particles is bonded to the amide nitrogen group of the compound having an amide group via the methylene group. A compound having an arbitrary substituent can be introduced as long as it is soluble in the organic solvent to be used.

  Since a compound having an amide group is introduced to the N-alkoxymethyl group of the present polymer particle, the unit derived from the N-alkoxymethyl (meth) acrylamide of the present polymer particle and the amide group derived from the introduced amide group The upper limit of the unit ratio is 1: 1 (molar ratio), and this ratio can be arbitrarily adjusted. The N-alkoxymethyl group that was not involved in the introduction of the amide group in the polymer particles is converted into a self-crosslinking structural unit represented by Chemical Formula 6 in the reaction for introducing the compound having an amide group.

化学式６中のＲ１はＣ４以上のアルキル基を表し、ｍは１以上の整数を表す。化学式６中のＡは重合体粒子を表す。

R1 in Chemical Formula 6 represents a C4 or higher alkyl group, and m represents an integer of 1 or higher. A in Chemical Formula 6 represents polymer particles.

  The present polymer particles into which an amide compound having a nonionic group as a substituent of the amide group of the compound to be introduced can be used as nonionic porous particles. Examples of amide compounds having nonionic groups include formamide, lactamide, acetate, acrylamide, gluconate, pantothenyl alcohol, benzoate, stearamide, cholic acid amide, polyacrylamide, polyamide resin, etc. Examples thereof include structural units derived from organic acid amides, amino sugar acetylated products such as N-acetoxyglucosamine and N-acetoxygalactosamine, urea, N-methylolurea, uridine and the like.

  In particular, a structural unit derived from a compound having a plurality of amide groups, such as urea, has a ratio of 1: 0.1 in the ratio of the N-alkoxymethyl group-derived unit of the polymer particles to the introduced amide group-derived amide group unit. To 1: 1 (molar ratio) by using within the range of not only high hydrophilicity but also porous particles having high mechanical strength due to cross-linking of the particles, a packing material for liquid chromatography Useful as. Such particles are particularly useful as particles having both high hydrophilicity and high mechanical strength, which are not present in conventional hydrophilic particles, as a chromatography carrier for desalting, and also have low swelling in organic solvents and the like. It is also useful as a normal phase chromatography support for polyol analysis.

  The polymer particles can easily introduce a compound having an amino group in which at least one bond on the nitrogen atom is an N—H bond in an organic solvent under an acid catalyst. The alkoxy group in the N-alkoxymethylamide group in the polymer particle is eliminated and the amide group nitrogen atom in the polymer particle is bonded to the amino nitrogen group of the compound having an amino group via the methylene group. It becomes an exchangeable particle. A compound having an arbitrary substituent can be introduced as long as it is soluble in the organic solvent to be used, and the ratio of the N-alkoxymethyl group-derived unit and the introduced amino group-derived amino group unit in the polymer particles The upper limit is 1: 1 (molar ratio), but the lower limit is not particularly limited. The N-alkoxymethyl group that is not involved in amino group introduction in the polymer particles is converted into a self-crosslinking structural unit represented by Chemical Formula 6 in the reaction for introducing the compound having an amino group. As an example of a compound having an amino group, a compound having an amide group can also be used. In this case, either one may have an N—H bond. Therefore, the introduction form includes the form in which the amino group is bonded to the amide group nitrogen atom in the polymer particle through a methylene group, and the form of the amide group in the polymer particle through the amide group nitrogen atom and methylene group. Introduced forms in which either or both forms of the combined form are present are mixed.

  The structural unit derived from the compound having an amino group introduced into the N-alkoxymethyl group in the polymer particles includes alkylamines such as ammonia, methylamine and ethylamine, and aromatics such as aniline, diaminobenzene and aminonaphthalene. Amino acids and their derivatives such as aromatic amines, taurine and alanine, arginine, asparagine, cysteine, glutamine, glycine, histidine, leucine, isoleucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, and the like Oligopeptides, adenine, cytosine, guanine and their derivatives, melamine, benzoguanamine, polyethyleneimine, polyallylamine, polyacrylamide, acrylamide and N, N-dimethyl List structural units derived from pigments such as lucifer yellow, acid red, and azofuroxine, such as a copolymer of ruaminopropylacrylamide and a copolymer of acrylamide and 2-acrylamido-2-methylpropanesulfonic acid. Can do.

  As a production method of the polymer particles, a general production method in which methylolation and etherification are performed from particles obtained by reverse-phase suspension polymerization of (meth) acrylamide with a water-soluble crosslinkable monomer is used. However, a method obtained by normal-phase suspension of N-alkoxymethylacrylamide and an oil-soluble crosslinkable monomer is preferable because the seed polymerization method can be used.

  As a polymerization initiator used for normal phase suspension polymerization, an oil-soluble polymerization initiator generally used for radical polymerization of vinyl monomers can be used. As an oil-soluble azo polymerization initiator, 2, 2 can be used. '-Azobis (4-methoxy-2,4-dimethylvaleronitrile), dimethyl-2,2'-azobis (2-methylpropionic acid), 1,1'-azobis (cyclohexane-1-carbonitrile), 2, Examples include 2′-azobis (2,4-dimethylvaleronitrile), 2,2′-azobis (isobutyronitrile), 2,2′-azobis (2-methylbutylnitrile), and oil-soluble organic peroxidation. 2,2-bis (4,4-di-t-butylperoxycyclohexyl) propane, 1,1-di (t-hexylperoxy) cyclohexane, 1,1 Di (t-butylperoxy) cyclohexane, di (3,5,5-trimethylhexanoyl) peroxide, dilauroyl peroxide, benzoyl peroxide, t-butylperoxyneodecanoic acid, t-hexylperoxypivalic acid, t-butylperoxypivalic acid, etc. Can be illustrated. In addition, since the polymerization is performed by suspension polymerization, it is preferable to use a polymerization initiator having a 10-hour half-life temperature of less than 100 ° C. The amount of the polymerization initiator used is in the range of 0.01 mol% to 1.0 mol%, particularly preferably in the range of 0.05 mol% to 0.5 mol%, based on the monomer. It is also possible to polymerize by ultraviolet rays or radiation without using a polymerization initiator.

  In the normal phase suspension polymerization in the present invention, a general polymer dispersion stabilizer used for stabilizing suspended oil droplets can be used. Examples thereof include polyvinyl alcohol, hydroxypropyl cellulose, polyvinyl pyrrolidone, polyethylene glycol, sodium polyacrylate, etc., but when using an organic acid polymer, the pH is set so that the aqueous phase does not become acidic. Therefore, it is preferable to use a general non-ionic polymer such as polyvinyl alcohol or hydroxypropyl cellulose. The amount used thereof can be 0.1 to 20% by weight as the concentration in the aqueous phase, and is preferably 0.5 to 10% by weight from the viewpoint of washing after polymerization. It is.

  Polymerization can be carried out by heating when using a thermally decomposable polymerization initiator, and by irradiation with radiation at room temperature when using radiation polymerization without using an initiator, and there is no particular limitation.

  In the case of normal phase suspension polymerization by seed polymerization, polystyrene and poly (styrene / acrylate ester) copolymer particles obtained by soap-free emulsion polymerization and dispersion polymerization methods used for the purpose of monodisperse particle size, For example, methacrylic acid ester particles can be used. In particular, it is preferable to use a seed particle polymer disclosed in JP-A-2001-2716 from the viewpoint of swelling property and the seed particle polymer hardly causing phase separation from the N-alkoxymethyl (meth) acrylamide polymer. In order to reduce the influence of the phase separation between the seed particle polymer and the polymer formed by polymerization of the monomer on the particle shape, the ratio of the seed particle polymer to the monomer is based on 1 part by weight of the seed particle. The monomer is preferably 10 parts by weight or more. Although there is no particular upper limit, for the purpose of narrowing the particle size distribution, the monomer content is preferably 10,000 parts by weight or less with respect to 1 part by weight of the seed particles. Therefore, there is no particular limitation on the swelling ratio represented by the ratio of the seed particle polymer to the monomer when seed polymerization is performed, and the swelling can be about 10,000 times the seed particle volume. It can be set according to.

  As the emulsifier used when carrying out normal phase suspension polymerization by seed polymerization, a general sulfonate or nonionic emulsifier can be used. Examples include nonionic emulsifiers such as alkyl sulfonates such as sodium lauryl sulfonate and sodium dodecylbenzene sulfonate, polyoxyethylene sorbitan stearate, polyoxyethylene stearyl ether, and polyoxyethylene sorbitan monolaurate. It can be illustrated. In particular, from the viewpoint of oil droplet stability, an alkyl sulfonate and a nonionic emulsifier of HLB 8 to 20 can be used in combination. The ratio of the nonionic emulsifier used in combination is 10% to 90% by weight, preferably 20% to 80% by weight, based on the alkyl sulfonate.

  When only alkyl sulfonate is used as an emulsifier used in normal phase suspension polymerization by seed polymerization, a compound having a higher hydrophobicity than N-alkoxymethyl (meth) acrylamide is used to improve oil droplet stability. It is also possible to use it. Examples of such highly hydrophobic compounds are C4 or higher alcohols such as butanol, pentanol, hexanol, cyclohexanol, octanol, acetic acid butyl ester, acetic acid pentyl ester, acetic acid hexyl ester, acetic acid phenyl ester, acetic acid benzyl ester. And organic acid esters such as propionic acid ethyl ester, propionic acid butyl ester, benzoic acid methyl ester and benzoic acid ethyl ester, and C6 to C9 aromatic compounds such as benzene, toluene and xylene. From the viewpoint of swelling, it is preferable to use higher alcohols having 5 to 6 carbon atoms or organic acid esters having -4 to 7 carbon atoms, benzene, and toluene. The amount used is 1% by weight or more based on the whole monomer, but it is not necessary to add more than necessary, and it is preferable to use 1% to 50% by weight based on the whole monomer.

  The polymer particles obtained by polymerization are washed with an appropriate solvent, generally warm water, in order to remove the polymer dispersion stabilizer.

  In the production method of the present invention, the obtained polymer particles are dispersed in an organic solvent, alkoxy groups are eliminated by an acid catalyst and bonded to other amide groups in the particles, thereby obtaining hydrophilic polymer particles. . In this step (hereinafter referred to as “second step”), it is also possible to introduce a compound having another amide group or amino group into the polymer particles using an acid catalyst. The polymer particles become hydrophobic by introducing a highly hydrophobic compound such as stearamide, and ion-exchange particles by introducing a compound having an ion-exchange group. In particular, by using an appropriate amount of a compound having a polyfunctional amide group or amino group, it is possible to simultaneously proceed with particle crosslinking.

  The organic solvent used in the second step in the production method of the present invention is preferably one that is stable to an acid catalyst and has no reactivity with an N-alkoxymethyl group, such as dioxane, tetrahydrofuran, benzene, toluene, xylene, dimethyl sulfoxide. , Higher alcohols such as dimethylformamide, butanol, pentanol, hexanol, acetic acid butyl ester, acetic acid pentyl ester, acetic acid hexyl ester, acetic acid phenyl ester, acetic acid benzyl ester, propionic acid ethyl ester, propionic acid butyl ester, benzoic acid methyl ester And organic acid esters such as ethyl benzoate. In addition, the organic solvent used in the second step can be a mixed solvent with water depending on the application. In particular, in order to make the polymer particles porous, it is preferable to carry out with a solvent in which the polymer particles swell. The amount used is 2 parts by weight, preferably 4 parts by weight or more, based on the polymer particles.

  The acid catalyst used in the second step in the production method of the present invention is not particularly limited, but strong acids are preferable, mineral acids such as hydrochloric acid, sulfuric acid and toluenesulfonic acid, and Lewis acids such as boron trifluoride are exemplified. it can. The amount used is not particularly limited, but is preferably 0.01% by weight or more, particularly preferably 0.1% by weight or more, based on the polymerized particles.

  In the second step of the production method of the present invention, by allowing a compound capable of reacting with an N-alkoxymethyl group such as an amide group or amino group having an N—H structure to coexist with the compound, Can be introduced in. Either one or both of the amide group and the amino group may be present, and it is necessary to have an N—H group to be introduced by elimination of the alkoxy group of the present polymer particle. Examples of compounds that can be introduced include formamide, lactic acid amide, acetic acid amide, acrylamide, gluconic acid amide, pantothenyl alcohol, benzoic acid amide, stearic acid amide, cholic acid amide, polyacrylamide, polyamide resin, and other organic acid amides, Amino sugar acetylated products such as N-acetoxyglucosamine and N-acetoxygalactosamine, alkylamines such as urea, N-methylolurea, uridine, ammonia, methylamine and ethylamine, and aromatic amines such as aniline, diaminobenzene and aminonaphthalene , Taurine and alanine, arginine, asparagine, cysteine, glutamine, glycine, histidine, leucine, isoleucine, lysine, methionine, phenylalanine, proline, serine, threonine, tri Amino acids such as tophan, tyrosine, valine and their derivatives, and their oligopeptides, adenine, cytosine, guanine and their derivatives, melamine, benzoguanamine, polyethyleneimine, polyallylamine, polyacrylamide, acrylamide and N, N-dimethyl Examples thereof include lucifer yellow, acid red, azofuroxin, and the like, such as a copolymer of aminopropyl acrylamide and a copolymer of acrylamide and 2-acrylamido-2-methylpropane sulfonic acid. The amount of the compound that can be introduced can be arbitrarily set in accordance with the purpose as long as it can be dissolved in an organic solvent.

  In the second step in the production method of the present invention, the reaction temperature and reaction time are appropriately adjusted according to the type of acid used and the amount used, and the degree of progress is measured by measuring the amount of alcohol produced by the elimination of the alkoxy group. Can be determined.

  The resulting crosslinked (meth) acrylamide particles can be easily removed of acid catalyst, organic solvent, residual introduced compound, etc. by washing with warm water or an organic solvent capable of dissolving the introduced amide group or amino group-containing compound. Is possible.

  According to the present invention, crosslinked (meth) acrylamide particles can be produced by normal phase suspension polymerization, and the particle diameter can be easily controlled. In particular, a seed polymerization method can be used, and it becomes possible to easily produce porous particles having a monodispersed particle size. In addition, the particles obtained by the present invention can easily have a highly crosslinked structure, and particles having mechanical strength much higher than those of polysaccharide particles and acrylamide particles produced by conventional reverse phase suspension polymerization. Manufacturable. In particular, crosslinked (meth) acrylamide particles having both hydrophilicity and mechanical strength can be set to a small particle size, and exhibit excellent performance as a packing material for liquid chromatography. In the second step of the present invention, an N-alkoxymethyl group, which is a functional group of the present polymer particle, can be bonded to an amide group, an amino group, or the like. Therefore, various compounds having these functional groups were introduced. It became possible to produce particles.

Hereinafter, the present invention will be described in detail with reference to examples, but the present invention is not limited thereto.

Example 1
<Manufacture of seed particles>
In a 500 mL three-necked flask, 30 g of benzyl methacrylate (manufactured by Tokyo Chemical Industry), 1.5 g of 2-ethylhexyl thioglycolic acid ester (manufactured by Tokyo Chemical Industry), 0.6 g of potassium persulfate (manufactured by Kishida Chemical) and Mill-Q water 250 g was added and a nitrogen purge was performed for 3 hours at room temperature with stirring. The flask was set in an oil bath set at 70 ° C., and polymerization was carried out for 6 hours. The agglomerates were separated by filtration to obtain seed particles.
The particle size of the seed particles was 1.1 μm CV 6.8% by SEM observation. Further, the molecular weight was measured by the size exclusion chromatography method (hereinafter referred to as SEC) with the following measuring apparatus, and as a result, it was 7800 in terms of polystyrene. The solid content concentration in the seed particle solution was 8.8%.

SEC measurement condition column: TSKgel GMHXL 7.8 mm ID × 30 cm (manufactured by Tosoh Corporation)
Eluent: Tetrahydrofuran feed pump: DP-8020 (manufactured by Tosoh Corporation)
Detector: UV-8020 wavelength set to 254 nm (manufactured by Tosoh)
Standard sample: Standard polystyrene (manufactured by Tosoh)
Measurement flow rate: 1.0 mL / min
Sample concentration: 1 mg / mL (THF solution)

<Polymerization of N-alkoxymethylacrylamide particles>
N-butoxymethylacrylamide (manufactured by Wako Pure Chemical Industries) 298.5 g, ethylene glycol dimethacrylate (manufactured by Tokyo Chemical Industry) 1.5 g, 1-pentanol (manufactured by Kishida Chemical) 60 g, V-65 (polymerization initiator Wako Pure Chemical Industries) 0.46 g) and 1.8 g sodium lauryl sulfonate (manufactured by Tokyo Chemical Industry) were weighed in a 1 L beaker and mixed well with stirring. To this, 600 mL of ion-exchanged water was added and emulsified using an ultrasonic homogenizer. To this, 0.182 g (solid content) of prepared seed particles, 600 mL of 4% polyvinyl alcohol aqueous solution (trade name POVAL 224, manufactured by Kuraray Industrial Co., Ltd.) was added and stirred well for 3 minutes.
The solution was transferred to a 3 L separable flask, a stirring blade was attached, and the mixture was slowly stirred at room temperature for 12 hours at a rotation speed of 100 rpm. Microscopic observation confirmed that the seed particles were swollen with the monomer mixture and swollen to about 13 μm.
The separable flask was placed in an oil bath set at 65 ° C., and polymerization was carried out for 3 hours while stirring. It was confirmed that the monomer had disappeared by gas chromatography analysis using an OV-17 (Gascro Industry) column. After completion of the polymerization, washing with warm water was performed to obtain polymer particles. The recovered weight was 477 g, and the water content was 41%.

<Second step>
500 mL Separa contains 85 g of hydropolymerized particles (dry weight 50 g), 9.4 g of urea (manufactured by Wako Pure Chemical Industries), 4.25 g of paratoluenesulfonic acid monohydrate (manufactured by Wako Pure Chemical Industries) and 340 g of dimethyl sulfoxide (manufactured by Kishida Chemical). The solution was put into a bull flask and stirred for 2 hours in an oil bath set at 90 ° C. The obtained hydrophilized particles were washed with warm water to obtain hydrophilic porous particles.

  The obtained hydrophilic porous particles were packed in a 6.0 mm ID × 15 cm column, and the pore characteristics were measured by size exclusion chromatography using polyethylene glycol (hereinafter referred to as PEG) as a standard sample. The hydrophilicity was compared with conventional hydrophilic particles using benzyl alcohol retention as an index. The mechanical strength was measured by a method in which pure water was passed in a state where the column was packed and the liquid feeding pressure was measured when the flow rate was changed linearly.

Pore property measurement condition column: 6.0 mm ID × 15 cm
Eluent: ion-exchanged water measurement flow rate: 0.5 mL / min
Liquid feed pump: DP-8020 (manufactured by Tosoh)
Detector: RI-8022 (manufactured by Tosoh)
Sample: ethylene glycol, polyethylene glycol (molecular weight 200, 300, 600, 1000, 3000, 6000, manufactured by Wako Pure Chemical Industries, Ltd.)
Exclusion limit molecular weight calculation method: PEG molecular weight PEG conversion molecular weight porosity calculation method at a linear molecular weight 6000 elution position passing through elution volumes of 200, 300 and 600: (ethylene glycol elution volume-PEG 6000 elution volume) / (column volume) -PEG6000 elution volume) x100
Hydrophobicity measurement conditions Samples other than measurement samples Same as pore characteristic measurement conditions: ethylene glycol, benzyl alcohol Hydrophobicity value: benzyl alcohol elution capacity / ethylene glycol elution capacity Mechanical strength (liquid feeding resistance) measurement condition column: 4.6 mm ID × 10 cm
Liquid solvent: Ion-exchanged water flow rate gradient: 0.2 mL / min to 10.2 mL / min for 10 minutes Linear gradient liquid pump: CCPM-II
Detection: Converted strength from liquid pump pressure gauge output: Estimated from liquid feeding pressure deviates from linear relationship with liquid feeding speed (inflection point) Table 1 shows the results of exclusion limit molecular weight, pore volume and hydrophobicity test. Shows the mechanical strength measurement results. Although the hydrophobicity was slightly large, the mechanical strength was very high although a slight pressure increase due to particle deformation was observed at around 8 MPa.

実施例２
＜Ｎ−アルコキシメチルアクリルアミド粒子の重合＞
Ｎ−ブトキシメチルクリルアミド（和光純薬製）２９４．０ｇ、エチレングリコールジメタクリレート（東京化成工業製）４．０ｇ、１−ヘキサノール（キシダ化学製） ３０ｇに変更した以外は実施例１と同様に重合体粒子を作成した。

＜第二工程＞
含水重合粒子１６．７ｇ（乾燥重量１０ｇ）、エチルアミン塩酸塩２．５ｇ（東京化成工業製）、６０％−硫酸水溶液１ｍＬ、ジメチルスルホキシド６０ｍＬ（キシダ化学製）を２００ｍＬ三つ口フラスコに投入し、９０℃に設定したオイルバス中で２時間撹拌した。得られた親水化粒子を温水洗浄しアニオン交換多孔質粒子１８．１ｇを得た。得られた粒子を０．５Ｎ−ＮａＯＨ水溶液で洗浄後、０．５Ｎ塩酸を用いて滴定を行ったところ、イオン交換容量は０．２５ｍｅｑ／（粒子−ｍＬ）であった。

Example 2
<Polymerization of N-alkoxymethylacrylamide particles>
N-butoxymethyl chloramide (Wako Pure Chemical Industries, Ltd.) 294.0 g, ethylene glycol dimethacrylate (Tokyo Chemical Industry Co., Ltd.) 4.0 g, 1-hexanol (Kishida Chemical Co., Ltd.) 30 g Polymer particles were made.

<Second step>
Water-containing polymer particles 16.7 g (dry weight 10 g), ethylamine hydrochloride 2.5 g (manufactured by Tokyo Chemical Industry Co., Ltd.), 60% -sulfuric acid aqueous solution 1 mL, dimethyl sulfoxide 60 mL (manufactured by Kishida Chemical) were put into a 200 mL three-necked flask, The mixture was stirred for 2 hours in an oil bath set at 90 ° C. The obtained hydrophilized particles were washed with warm water to obtain 18.1 g of anion exchange porous particles. The obtained particles were washed with 0.5N-NaOH aqueous solution and titrated with 0.5N hydrochloric acid. As a result, the ion exchange capacity was 0.25 meq / (particles-mL).

Example 3
The following second process was performed using the N-alkoxymethylacrylamide particles prepared in Example 2.
<Second step>
Water-containing polymer particles 16.7 g (dry weight 10 g), taurine 2.25 g (manufactured by Wako Pure Chemical Industries), 60% -sulfuric acid aqueous solution 1 mL, dimethyl sulfoxide 55 mL (manufactured by Kishida Chemical), and distilled water 5 mL are charged into a 200 mL three-necked flask. The mixture was stirred for 2 hours in an oil bath set at 90 ° C. The obtained hydrophilic particles were washed with warm water to obtain 16.1 g of ion-exchange porous particles having a betaine structure. The obtained particles were washed with 0.5 N HCl and titrated with 0.1 N NaOH, and the ion exchange capacity was 0.08 meq / (particles-mL).

Example 4
Using the N-alkoxymethylacrylamide particles prepared in Example 2, the same procedure as in Example 2 was carried out except that the second step was changed to 2.5 g of glycine and 2 mL of 60% sulfuric acid aqueous solution as the amino compound. The obtained particles were washed with 0.5N HCl and titrated with 0.1N NaOH. The ion exchange capacity was 0.07 meq / (particles-mL).

Example 5
<Creation of acrylamide / 2-acrylamido-2-methylpropanesulfonic acid copolymer>
3 g of acrylamide (manufactured by Aldrich), 7 g of 2-acrylamido-2-methylpropanesulfonic acid (manufactured by Aldrich), and 0.1 g of potassium persulfate (manufactured by Kishida Chemical) are dissolved in ion-exchanged water and put into a 100 mL flask to form nitrogen. Polymerization was carried out at 70 ° C. for 16 hours under an air stream.

<Second step>
Subsequently, Example 3 was used except that the N-alkoxymethylacrylamide particles prepared in Example 2 were used and the second step was changed to 5 mL of an acrylamide-2-acrylamide-2-methylpropanesulfonic acid copolymer solution as an amide compound. Manufactured in the same manner. The obtained particles were washed with 0.5 N HCl and titrated with 0.1 N NaOH, and the ion exchange capacity was 0.013 meq / (particles-mL).

Example 6
The particles prepared in Example 1 were packed in a 10 mm ID × 15 cm liquid chromatography column and subjected to desalting chromatography. Chromatographic conditions are shown below.
Eluent: 50 mmol / L phosphate buffer pH 6.5
Measurement flow rate: 3 mL / min
Temperature: 4 ° C
Sample: 10 mg / mL Ovalbumin (manufactured by Sigma) 0.5 mol / L NaCl and 50 mmol / L phosphate buffer pH 6.5
Sample volume: 1 mL, 2 mL, 3 mL, 4 mL
apparatus:
Liquid feed pump DP-8020 (manufactured by Tosoh)
Column oven CO-8020C (manufactured by Tosoh)
Detector UV-8020 (manufactured by Tosoh) Gradient monitor GM-8010 (manufactured by Tosoh) set to a wavelength of 280 nm
The obtained chromatogram is shown in FIG. Protein and salt could be separated efficiently.

Example 7
The particles prepared in Example 1 were packed in a 4.6 mm ID × 15 cm liquid chromatography column, and normal phase chromatography, which is a polyol analysis method, was performed. Chromatographic conditions are shown below.
Eluent:
1) Acetonitrile / water = 7/3
2) Acetonitrile / 5 mmol / L phosphate buffer pH 6.5 = 7/3
Measurement flow rate: 1 mL / min
Temperature: 80 ° C
sample:
Eluent 1) Ethylene glycol, glycerol, sorbitol, glucose, saccharose, maltose Eluent 2) Ethylene glycol, glycerol, glucose, glucosamine, galactoconic acid apparatus:
Liquid feed pump DP-8020 (manufactured by Tosoh)
Column oven CO-8020C (manufactured by Tosoh)
Detector RI-8022 (manufactured by Tosoh Corporation)
The obtained chromatogram is shown in FIGS. The polyol was separable efficiently. In addition, when conventional silica-based fillers are used, simultaneous analysis of basic sugar, neutral sugar and acidic sugar, which is difficult to perform simultaneous analysis, is possible.

Comparative Example 1
PIERCE polyacrylamide product name D-Salt Polyacrylamide 1800 was extracted from the column, and the same measurement as in Example 1 was performed. Although the pore volume is large and the hydrophobicity is small, the particle strength is large, so that the liquid feeding resistance is small, but the mechanical strength is weak and the material is crushed at about 1.7 MPa.

Comparative Example 2
Product name manufactured by Amersham Sephadex G25 Superfine The same measurement as in Example 1 was performed using crosslinked dextran particles. Although the pore volume is large and the hydrophobicity is small, the particle diameter is large, so that the liquid feeding resistance is small, but the mechanical strength is weak and it is crushed at about 0.95 MPa.

  Since the polymer particles of the present invention can introduce various compounds having an amide group or amino group or both, various binders, coating materials, cosmetics, immunodiagnostic carriers, packing for liquid chromatography Usage as an agent is conceivable. In particular, polymer particles having characteristics of having both low hydrophobicity and high mechanical strength have particularly excellent performance as a packing material for liquid chromatography.

Indicates the change in the liquid supply pressure when the flow rate of liquid sent to the column is changed. Results of desalting chromatography of protein solution containing salt Chromatographic results of neutral polyols Simultaneous chromatographic results for basic, neutral and acidic sugars

Claims (15)

A unit derived from an N-alkoxymethyl (meth) acrylamide monomer represented by Chemical Formula 1 (X in Chemical Formula 1 is hydrogen or a methyl group, and R1 represents a C4 or higher alkyl group);
In a polymer particle having a particle diameter of 1 to 1000 μm and including a unit derived from a polyfunctional unsaturated monomer that can be copolymerized with the monomer represented by Chemical Formula 1, N − represented by Chemical Formula 1 A unit derived from an alkoxymethyl (meth) acrylamide monomer is represented by chemical formula 2 (wherein R2 in chemical formula 2 is H, C3 or lower alkyl group or C3 or lower and an alkyl group substituted with one or more hydroxyl groups). R3 in Chemical Formula 2 is H, NH2, a C1-C18 alkyl group, a phenyl group, a linear, branched or cyclic alkyl group which is C24 or less and substituted with one or more hydroxyl groups, -CH2- Represents any one of CO-NH2, or R3 is a structural unit derived from another N-alkoxymethyl (meth) acrylamide monomer, and other N-alkoxymethyl ( A) in the chemical formula 2 represents a polymer particle) and / or chemical formula 3 (wherein R4 and R5 in the chemical formula 3 are H, C6 or lower alkyl group, phenyl group, naphthyl group, C6 or lower alkyl group substituted with one or more hydroxyl groups, -CH2COOH, -CH2SO3H, -CH2CH2SO3H,-(CH2) n-PO- (OH) 2 (Wherein n represents an integer of 1 to 4), -CHR6-PO- (OH) 2 (wherein R6 represents a linear alkyl group having 1 to 3 carbon atoms), -CHR7-COOH (R7 is an amino acid) Represents a side chain), oligopeptide residue, adenine residue, cytosine residue, guanine residue, melamine residue, benzoguanamine residue, polyethyleneimine, polyallylamine Alternatively, R4 and R5 are structural units derived from other N-alkoxymethyl (meth) acrylamide monomers, and are derived from other N-alkoxymethyl (meth) acrylamide monomers via a methylene group. Cross-linked (meth) acrylamide particles into which at least the substituent represented by A in Chemical Formula 3 represents a polymer particle is introduced.

The molar ratio of the unit derived from the N-alkoxymethyl (meth) acrylamide monomer represented by Chemical Formula 1 and the unit derived from the polyfunctional unsaturated monomer is 99.99: 0.01 to 50:50. The crosslinked (meth) acrylamide particles according to claim 1, which are within a range. The crosslinked (meth) acrylamide particle according to claim 1 or 2, wherein the polyacrylamide polymer or the copolymer containing polyacrylamide is bonded to the main chain of the polymer particle. 2. At least one of R3, R4 and R5 according to claim 1 contains at least one ion exchange group selected from a sulfonic acid group, a carboxylic acid group, a phosphoric acid group and an amino group. 3. Crosslinked (meth) acrylamide particles according to 3. 5. The crosslinked (meth) acrylamide particles according to claim 1, wherein the polymer particles have a pore diameter of 5000 or less with polyethylene glycol as a sample. A method for producing crosslinked (meth) acrylamide particles according to any one of claims 1 to 5,
(1) Oil-soluble N-alkoxymethyl (meth) acrylamide represented by Chemical Formula 1, a polyfunctional unsaturated monomer copolymerizable with the monomer represented by Chemical Formula 1, and a suspension stabilizer in water A step of obtaining polymer particles by suspending in a phase and polymerizing,
(2) The compound having an amide group or amino group is dispersed or dissolved, and the acid catalyst is allowed to act in the organic solvent in which the polymer particles in the step (1) are dispersed, thereby deriving from the monomer of the chemical formula 1. Introducing a substituent represented by Chemical Formula 2 and / or Chemical Formula 3 into the unit;
A method for producing crosslinked (meth) acrylamide particles comprising: 7. The method for producing crosslinked (meth) acrylamide particles according to claim 6, wherein in step (1), an oil-soluble N-alkoxymethyl (meth) acrylamide represented by Chemical Formula 1 and a single amount represented by Chemical Formula 1 A mixture containing a polyfunctional unsaturated monomer that can be copolymerized with the body is emulsified in water using an emulsifier, and the seed polymer particles are allowed to act on this to cause the seed polymer particles to absorb the monomer and monodisperse the particle size A method for producing crosslinked (meth) acrylamide particles, wherein monomer oil droplets are prepared and polymerized. In the method for producing crosslinked (meth) acrylamide particles, at least one or more selected from C4 or higher alcohols or organic acid esters and C6 to C9 aromatic compounds as stabilizers for the monodisperse monomer droplets having a particle size of The method for producing crosslinked (meth) acrylamide particles according to claim 7, wherein the organic solvent is allowed to act on the monomer in an amount of 1% by weight or more. The crosslinked (meth) acrylamide particles according to claim 7 or 8, wherein the emulsifier is an alkyl sulfonate, an alkyl benzene sulfonate, or an alkyl ether sulfate, and is used in an amount of 0.1 to 5% by weight based on the monomer. Manufacturing method. The method for producing crosslinked (meth) acrylamide particles according to claim 9, wherein a nonionic emulsifier of 1 to 50% by weight of HLB 8 to 20% is used in combination with the emulsifier. The acid catalyst is selected from mineral acids such as hydrochloric acid, phosphoric acid and sulfuric acid, alkylsulfonic acid, benzenesulfonic acid, toluenesulfonic acid, organic acids such as oxalic acid, acetic acid and maleic acid, and Lewis acids such as trifluoroboron. The method according to any one of claims 6 to 10, which is one or more acids. The compound having an amide group or amino group is a compound selected from the group consisting of formamide, urea, N-methylolurea, acetamide, lactamide, 2-hydroxypropionamide, oxamide, malonamide, and polyacrylamide. The method for producing crosslinked (meth) acrylamide particles according to any one of 10 above. A packed column in which the crosslinked (meth) acrylamide particles according to any one of claims 1 to 5 are packed in a liquid chromatography column. A method of using the crosslinked (meth) acrylamide particles according to any one of claims 1 to 3 or claim 5 as a desalting column after being packed in a liquid chromatography column. A method in which the crosslinked (meth) acrylamide particles according to any one of claims 1 to 3 are packed in a liquid chromatography column and used as a polyol analysis column.

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