JPS6384641A - Strongly acidic cation-exchange resin and its production - Google Patents

Abstract

PURPOSE:To increase the mechanical strength of the title cation exchange resin by bonding a specified atomic group to the glycidyl group of a gelled copolymer obtained by cross-linking glycidyl monovinyl ester with the polyvinyl ester of a polyhydric alcohol. CONSTITUTION:The ring of the glycifyl group based on (A) the glycidyl monovinyl ester of a cross-linked gelled copoplymer from the (A) component and (B) the polyvinyl ester of a polyhydric alcohol is opened and modified with water, etc., to form a hydroxyl group, and propane sultone, etc., are allowed to act on the hydroxyl group. The atomic group expressed by formula I [X is a propylene group or a butylene group, (l) is o or 1, (m) is 2-4 integer, and (m) is 0 when (l) is 1] is bonded to the obtained material to produce a strongly acidic cation-exchange resin consisting of a porous polymer. When the resin is used as the packing material for high-speed liq. chromatography, the eluate can be sent at a high speed.

Description

Detailed Description of the Invention "Industrial Field of Application" The present invention is directed to a high-performance liquid chromatography agent that has excellent mechanical strength and chemical stability, and exhibits small volume changes during use, such as an aqueous solvent-based high-performance liquid chromatography agent. The present invention relates to a hydrophilic, strongly acidic cation exchange resin suitable for use as a hydrophilic, strongly acidic cation exchange resin, and a method for producing the same.

[Prior Art] Recently, separation of proteins, enzymes, etc. using ion exchange chromatography has become popular. For example, strongly acidic cation exchangers, such as cross-linked dextran and cross-linked agarose into which sulfone groups have been introduced, are used as packing materials for liquid chromatography for the purpose of separating and analyzing biological components. However, all of these ion exchangers have extremely high swelling degrees and low mechanical strength, so
It has the disadvantage that it cannot be used in applications that use packing materials with small particle sizes, such as high-performance liquid chromatography packing agents.

Ion exchangers of acrylic acid or methacrylic acid and divinylbenzene copolymers are also known. However, this copolymer is not preferable because hydrophobic adsorption of methacrylic acid and novinylbenzene in the skeleton and hydrophobic adsorption of biological components may occur.

Furthermore, ion exchangers in which ion exchange groups are bonded to the surface of silica gel are also known. However, silica gel has the disadvantage that it dissolves under slightly alkaline conditions.

[Problems to be Solved by the Invention] The object of the present invention is to overcome the drawbacks of the conventional ion exchangers and to provide a hydrophilic material with high mechanical strength, excellent chemical stability, and low swelling degree. The present invention provides a strongly acidic cation exchange resin and a method for producing the same.

[Means for Solving the Problems] According to the present invention, a hydrophilic strongly acidic cation exchange resin that satisfies the above objects and a method for producing the same are provided.

That is, the present invention comprises (A) glycidyl monovinyl ester or glycidyl monovinyl ether and (B) polyvinyl ester of polyhydric alcohol as main components;
) is a gel-like copolymer (hereinafter referred to as the gel-like copolymer according to the present invention) crosslinked with the (B) component, and the glycysol group based on the above-mentioned (4) component has the general formula (I). H (wherein, X represents a 2-hydroxypropylene group, a propylene group or a butylene group; t represents 0, 1 or 2; m represents O or an integer from represents an integer, p represents 0 or 1, p
When is O, t and m are 0, and when X is a 2-hydroxypropylene group, when t is 0, m is O, and when t is 1
When m is not O, and when X is a propylene group or a butylene group, t is O or 1, and when t is 1, m is 0.
It is. ) This invention relates to a strongly acidic cation exchange resin characterized by being made of a porous copolymer to which atomic groups represented by the following are bonded.

In addition, the present invention provides the method of causing propane sultone or 1,4-butane sultone to act on the hydroxyl group generated by ring-opening modification of the glycidyl group of the gel copolymer according to the present invention with water, glycerin or polyalkylene glycol,
The hydroxyl group has the general formula (II) (wherein, X represents a propylene group or a butylene group, represents til-io or 1, m represents O'' or an integer from 1 to 15, and n
represents an integer from 2 to 4, and when t is 1, m is O. This invention relates to a method for producing a strongly acidic cation exchange resin, which is characterized by bonding atomic groups represented by ) to obtain a porous copolymer.

Furthermore, the present invention provides epihalohydrin, ethylene glycol diglycidyl ether, propylene, etc. in the glycysol group of the gel copolymer according to the present invention, or in the hydroxyl group generated by ring-opening modification of the glycidyl group with water or glycerin. Glycidyl of a modified gel-like copolymer obtained by combining glycol didali cisole ether, 1,3-pro-yanool no glycidyl ether, 1,4-butanol no no glycidyl ether or polyethylene glycol sogellicidyl ether A sulfite compound is applied to the glycidyl group to form the general formula (2) or 1 to 15.
n represents an integer from 2 to 4,
p represents O or 1; when p is O, t and m are 0; when L is O, m is O; when t is 1, m is not O; This invention relates to a method for producing a strongly acidic cation exchange resin, which is characterized by bonding atomic groups represented by ) to obtain a porous copolymer.

Hereinafter, the hydrophilic strongly acidic cation exchange resin of the present invention and its manufacturing method will be explained.

The gel-like copolymer according to the present invention is disclosed in JP-A-53-1087
As described in the publication, for example, aqueous suspension polymerization of glycidyl monovinyl ester or glycidyl monovinyl ethyl and (B) polyvinyl ester of a polyhydric alcohol as a crosslinking agent in the presence of a diluent. It can be synthesized by

As component (A), glycidyl monovinyl ester,
Glycidyl acrylate, glycidyl methacrylate, etc. are used. Further, as the glycidyl monovinyl ether of component (A), allyl glycidyl ether, meta-allyl glycisol ether, etc. are used.

On the other hand, as the polyvinyl ester of polyhydric alcohol as the component (B) as a crosslinking agent, ethylene glycol diacrylate, ethylene glycol dimethacrylate, propylene glycol diacrylate, propylene glycol dimethacrylate, glycerin diacrylate, glycerin dimethacrylate, glycerin triacrylate, etc. Acrylate, glycerol trimethacrylate, pentaerythritol soacrylate, intererythritol dimethacrylate, R-intererythritol trimethacrylate, flubitol dimethacrylate, and sorbitol trimethacrylate.

In the present invention, component (A) may be used as the main component, and other monomers copolymerizable therewith may be appropriately used in combination.

(B) Component polyvinyl ester of polyhydric alcohol is
It is used in an amount of 5 to 90 mol of the total polymerizable monomer, preferably 10 to 60 mol of the total polymerizable monomer.

The diluent used during polymerization is a solvent for component (A) and component CB) that is inert to the polymerization reaction. Further, the diluent needs to be insoluble or sparingly soluble in water, which is a polymerization dispersion medium.

Such diluents include halodanized hydrocarbons such as chlorobenzene, dichloromethane, dichloroethane, trichloroethane; aliphatic or aromatic esters such as ethyl acetate, propyl acetate, butyl acetate, amyl acetate; butyl alcohol, amyl acetate, etc. Alcohol, octyl alcohol, lauryl alcohol, cyclohexanol, etc.:
! - I can list the types.

The amount of diluent used is in the range of 20 to 200 parts by weight based on 100 parts by weight of the total amount of component (4) and component (B) used. If the amount of the diluent used is less than 20 parts by weight, the pore volume of the resulting gel-like copolymer according to the present invention will decrease, resulting in a decrease in separation performance, and if it is more than 200 parts by weight, the gel-like copolymer according to the present invention will Copolymer lacks mechanical strength.

The initiator used in the polymerization may be a general radical polymerization initiator used in normal suspension polymerization, such as 2
, 2'-azobisisobutyronitrile, 2,2'-azohis-(2,4-trimethylvaleronitrile), and peroxide-based initiators such as benzoyl peroxide and lauroyl peroxide. Initiators can be used.

When performing suspension polymerization, it is preferable to add a dispersion stabilizer such as polyvinyl alcohol, carboxymethyl cellulose, or gelatin to the aqueous phase, and also to ensure that polymerizable monomers such as components (A) and (B) are polymerized. In order to prevent the dispersion medium from dissolving in water, salts such as sodium chloride, sodium sulfate, calcium chloride, etc. may be dissolved in water.

The amount of the aqueous layer used is from approximately the same volume or more to about 10 times the volume of the organic layer.

The polymerization reaction is usually carried out at 50 to 90°C for 3 to 16 hours. After completion of polymerization, the copolymer is filtered, washed with water, and further washed with an organic solvent.

The gel-like copolymer particles according to the present invention thus obtained are porous and have an average pore diameter of 100X to 2000X, and are spherical particles with an average particle diameter of several microns to 100 microns. It is.

As a method for bonding the atomic group represented by the general formula (I) to the glycidyl group of the gel copolymer according to the present invention, (a) A method of ring-opening modification of a glycidyl group with water, glycerin or polyethylene glycol, and bonding a sulfoalkyl group to the generated hydroxyl group; (b) a glycidyl group based on component (A) of the gel copolymer according to the present invention; Examples include a method of bonding a sulfone group to the glycidyl group of the modified gel copolymer according to the present invention, and a method of bonding a sulfone group to the glycidyl group of the modified gel copolymer according to the present invention.

In the method (a), the ring-opening modification with water is carried out by heating the gel-like copolymer according to the present invention in water at 70° C. to 100° C. for 2 to 10 hours using sulfuric acid or hydrochloric acid as a catalyst. The concentration of acid used as a catalyst is 0.05~
1 regulation is preferred.

Ring-opening modification with glycerin or polyethylene glycol is performed by adding the gel-like copolymer according to the present invention in glycerin or polyethylene glycol, or in an organic solvent containing glycerin or polyethylene glycol, using boron trifluoride etherate as a catalyst, using a conventional method. 40-80
This is done by heating for 1 to 5 hours at .

The sulfoalkyl group is introduced by reacting the ring-opening modified gel copolymer with propane sultone or 1,4-butane sultone in an organic solvent using an alkali metal hydroxide as a catalyst. The reaction temperature and reaction time are usually room temperature to 100°C for 2 to 24 hours.

In addition, the introduction of sulfone groups in the method (b) is carried out by allowing a sulfite compound to act on the glycidyl groups of the gel-like copolymer according to the present invention in an aqueous solution.

Furthermore, the introduction of sulfone groups in the method (c) is carried out by allowing a sulfite compound to act on the glycidyl groups of the modified gel-like copolymer according to the present invention in an aqueous solution.

In the method (c), the modified gel copolymer is obtained by ring-opening modification of the gel copolymer according to the present invention with water or glycerin, and then ring-opening modification to produce hydroxyl group niepihalohydrin or ethylene glycol diglycidyl. Ether, propylene glycol diglycidyl ether, 1.3-7
It can be synthesized by combining compounds having a glycidyl group such as lopanediol cyclidyl ether, 1,4-butanediol diglycidyl ether, or polyethylene glycol diglycidyl ether. Ring-opening modification of the glycidyl group of the gel-like copolymer according to the present invention with water or glycerin is carried out in the same manner as in the case (a) above. The method for introducing the above compound having a glycysol group into a ring-opening modified gel copolymer is as follows:
Although various methods can be employed, two typical methods are shown below.

In the first method, a ring-opening modified gel copolymer is reacted with epichlorhydrin, ebibromohydrin, or the like in an aqueous solution of an inorganic base such as sodium hydroxide or potassium carbonate.

The reaction temperature is room temperature to 80°C, and the reaction time is 1 to 8 hours.

The second method involves adding ethylene glycol sogelicidyl ether, propylene glycol diglycidyl ether, propylene glycol diglycidyl ether, This is carried out by reacting 1.3-7'ropanol noglycidyl ether, 1.4-7" tandiol diglycidyl ether, or polyethylene glycol diglycidyl ether. The polyethylene glycol diglycidyl ether used is polyethylene glycol Repeating unit of diglycy sol ether -C2H4
It is preferable to use one having a repeating number of 0- from 1 to 10.

The concentration of the alkaline aqueous solution is usually preferably 1 to 10 normal. The reaction temperature is room temperature to 100℃, and the reaction time is 1
~24 hours.

The sulfite compounds used in the method (b) and the method (b) include sodium sulfite, sodium hydrogen sulfite,
Potassium sulfite, ammonium sulfite, etc. The concentration of the sulfite compound is preferably 1 mol to 5 mol/l. The reaction temperature is room temperature to 100°C, and the reaction time is 2 hours to 24 hours.

[Effects of the Invention] The hydrophilic strongly acidic cation exchange resin obtained by the present invention has the following advantages compared to conventional ion exchangers.

1) It has high mechanical strength, and when used, for example, as a high-performance liquid chromatography stimulant, the eluent can be fed at high speed, making rapid analysis possible.

2) Since the degree of crosslinking is high, the degree of swelling is low and it can be used at high salt concentrations.

3) It is stable over a wide pH range, and can be used stably even under alkaline conditions, which cannot be applied to fillers using silica gel as a carrier, for example.

4) Since the resin surface is covered with hydrophilic hydroxyl groups and ether groups, it has sufficient hydrophilicity, and when separating biological components etc., hydrophobic adsorption is small.

[Examples] Hereinafter, the present invention will be explained in more detail with reference to Examples, but the present invention is not limited to the following Examples unless it exceeds the gist thereof.

Example 1 (1) Production of gel copolymer particles having glycidyl groups A mixture of glycidyl methacrylate 500F, ethylene glycol dimethacrylate 20ON, n-dotecanol 150g, chlorobenzene 800g and azobisisobutyronitrile 10.9g was prepared. , 15.17 was added to an aqueous solution of 61 in which polyvinyl alcohol of 15.17 was dissolved, and suspension polymerization was carried out at 60° C. for 8 hours while stirring at high speed.

After cooling the reactant, the generated copolymer particles were collected by filtration,
Washed with water. Next, this copolymer cake was placed in 4 tons of toluene, stirred at room temperature for 2 hours, and then filtered. Further, this copolymer cake was added to 46 denatured alcohol and stirred for 1 hour. After filtering this copolymer, it was washed with denatured alcohol 21 and dried at 80° C. for 8 hours. The copolymer particles subjected to the above operations were classified to obtain 210 g of a gel-like copolymer having a glycidyl group with a particle size of 10 to 15 microns.

(2) 10 g of gel-like copolymer particles obtained in ring-opening (1) with water was added to 0.5
The mixture was well dispersed in a specified aqueous sulfuric acid solution (100 mA') and maintained at 90° C. for 4 hours while stirring. After cooling, the reaction product was collected by filtration and washed with water. No free epoxy groups were found in the reactant cake.

(3) Introduction of sulfopropyl groups After drying the ring-opening modified gel copolymer particles obtained in (2), 10 g of the copolymer particles were taken and added with 100 ml of dimethyl sulfoxide. ;y' Ronohira Nonsulton 15
5 ml of sodium I,20 hydroxide was added, and the mixture was reacted at 30°C for 7 hours. After the reaction was completed, the particles were washed with water. The ion exchange capacity of the obtained particles was measured by neutralization titration and was found to be 0.63 mmol/? -The presence of a sulfopropyl group in drygel was observed.

Example 2 (4) Ring-opening with glycerin (1) Disperse the R-shaped copolymer particles 10F obtained in step (1) in a mixed solution of 5Qml of dioxane and 5Qml of glycerin, and add 0.5ml of boron trifluoride niteller). In addition, the mixture was stirred at 70° C. for 4 hours to carry out an addition reaction of glycerin to glycidyl groups. The obtained copolymer particles were thoroughly washed with acetone and water to obtain a copolymer cake of 301. No free epoxy groups were found in the copolymer cake.

(5) After drying the ring-opening modified gel-like copolymer particles obtained by introduction of sulfopropyl groups (4), take copolymer particles ION, and use sulfopropyl groups in exactly the same manner as in (3). The group was introduced. As a result, an ion exchange resin having an ion exchange capacity of 0.53 mmol/ji-drygel was obtained.

Example 3 (6) 10 g of gel-like copolymer particles obtained in ring-opening with polyethylene glycol (1) were mixed with dioxane 5Qm.
A' and polyethylene glycol 200 (average molecular weight 2
00) 50m7? 0.5 ml of boron trifluoride Nieteller) was added thereto, and the mixture was stirred at 70°C for 4 hours to carry out an addition reaction of polyethylene glycol to glycidyl groups.

The obtained copolymer particles were thoroughly washed with acetone and water,
30 g of copolymer cake was obtained.

(7) Introduction of sulfopropyl groups After drying the copolymer cake obtained in (6), 10 g of copolymer particles were taken, and sulfopropyl groups were introduced in the same manner as in (3). Result, 0.45
An ion exchange resin with an ion exchange capacity of mmol/g-drygel was obtained.

Example 4 (8) Introduction of glycidyl groups with epichlorohydrin To 30 g of cake of the ring-opening modified gel copolymer particles obtained in (2), 60 ml of 2N sodium hydroxide and 20 TL of epichlorohydrin were added, and the mixture was heated at 50°C. Stirred for 2 hours. The reaction product was washed with water and acetone and then dried at room temperature. A portion of this copolymer was taken and reacted with 1.3 mol of sodium thiosulfate, and the produced OH- was titrated to quantify the glycidyl group (J., Porras, Journal of Chromatography Chiku 90). 87-98, 1974).

As a result, the presence of glycidyl groups at 0.51 mmol/g-dryge 1 was observed.

(9) Introduction of sulfone groups To the modified gel copolymer 10.9 obtained in (8), add 60 rnl of 25 thiosulfite aqueous solution and mix at 70°C.
The mixture was stirred for 8 hours. Next, the obtained reaction product was washed with water.

One part of the reaction product was taken and the amount of sulfone group was determined by neutralization titration. As a result, 0.35 mmol/fI-dry
Sulfone groups of gel were observed.

Example 5 Cake 3 of ring-opening modified copolymer particles obtained by introducing glycidyl groups with α0 1,4-butanediol diglycidyl ether (2)
0! ! 30 ml of 1N sodium hydroxide, 60 ml of sodium borohydride and 30 ml of 1,4-butanediol diglycidyl ether were added to the mixture, and the mixture was heated at 30°C with stirring.
It was held for 5 hours. The reaction product was taken, washed with acetone, water, and acetone, and dried at room temperature.

When the glycidyl groups in the reaction product were quantified in the same manner as in (8), 0.30 mmol/9-drygel of glycidyl groups was observed.

(11) Introduction of sulfone group to 10 g of modified gel-like copolymer particles obtained by αQ (9)
A sulfone group was introduced in a similar manner to Tobaritoku. 0
．． 25 mi! An ion exchange resin was obtained having an ion exchange capacity of J mol/9-drygel.

Applied gAJ 1 Fill a stainless steel column (inner diameter 8 fins x length 75 fins) with the cation exchange resin obtained in Jitsugi (3), and use 20 mmol phosphate buffer (pH 7.0) as the eluent for elution. 20 mmol phosphate buffer + 0.5 molar sodium chloride (pH 7.0) was used as solution (B), flow rate Q, 5 ml 17 ml
Elution 1 (a 40-minute linear gradient from A to eluent (B) was carried out, and a mixture of ■ myoglobin, ■ trinosinoke, ■ ribonuclease A1, ■ α-chymotrypsinodan A1, ■ cytochrome C1, and ■ lysozyme were obtained. When analyzed, we were able to separate six proteins as shown in Figure 1.

As a comparative example, a stainless steel column was similarly filled with the copolymer having no cation exchange group, which had been ring-opened by polymerization water removal obtained in Example 1-(2), and was treated under the same conditions as above. A mixture of (1) myoglobin, (2) trypsinogen, (2) ribonuclease A1, (2) α-chymotrypsinodan A1, (3) cytochrome C10, and lysozyme was analyzed, but the six components leaked out almost simultaneously and could not be separated.

Furthermore, examples! - Add 1N saline solution at 1ml/mi to the column filled with the ion exchange resin obtained in (3).
It was sent for 20 hours at a flow rate of n. In this case, the column pressure is 10
kg/crn2, and no increase in column pressure was observed during liquid feeding. Also, when I opened the top of this column,
No gaps due to shrinkage were observed.

Next, the ion exchange resin 5 obtained in Example 1-(3)
g of 0.01N aqueous sodium hydroxide solution 100m7
! After putting it in the container and leaving it at room temperature for 50 hours, the ion exchange capacity was measured and found to be 0.64 meq/ji.
No change in ion exchange capacity was observed before immersion in the sodium hydroxide aqueous solution. These results indicate that the cation exchange resin of the present invention is extremely stable even in alkaline solutions.

Application example 2 The cation exchange resin obtained in Example 5-αη was packed into a stainless steel column (inner diameter 8 mm x length 75 mm), and myoglobin and lysonuclease A1 were analyzed under the same analytical conditions as in application example 1. When the mixture of α-chymotrypsinokone A1 and cytochrome C1 was analyzed, it was possible to separate five tannophosphates as shown in FIG. 2.

[Brief explanation of the drawing]

FIG. 1 is a chromatogram obtained when a protein mixture was subjected to ion exchange chromatography using the cation exchange resin of the present invention obtained in Example 1-(3). In the figure, ■ is myoglobin, ■ is trigysinodan, ■ acupuncture ponuclease A1, and ■ is α-chymotrypsinodan A.
, ■ indicate each peak of cytochrome C, d and lysozyme. FIG. 2 is a chromatogram obtained when a protein mixture was subjected to ion exchange chromatography using the cation exchange resin of the present invention obtained in Example 5-α. In the figure, ■ is myoglobin, ■ is bonuclease-A, and ■ is
indicates the peaks of α-chymotrypsinoke 9-AX, cytochrome C1, and pyrozozyme.

Claims (3)

[Claims] (1) The main components are (A) glycidyl monovinyl ester or glycidyl monovinyl ether and (B) polyvinyl ester of polyhydric alcohol, and the component (A) is (
The glycidyl group based on the component (A) of the gel copolymer crosslinked with component B) has the general formula (I)▲mathematical formula, chemical formula, table, etc.▼...(I) (in the formula, X represents a 2-hydroxypropylene group, propylene group or butylene group; l represents 0, 1 or 2; m represents 0 or an integer from 1 to 15; n represents an integer from 2 to 4; p represents 0 or 1, p
When is 0, l and m are 0, when X is 2-hydroxypropylene group, when l is 0, m is 0, and when l is 1
When , m is not 0, when X is a propylene group or a butylene group, l is 0 or 1, and when l is 1, m is 0
It is. ) A strongly acidic cation exchange resin characterized by being composed of a porous copolymer in which atomic groups represented by the following are bonded. (2) The main components are (A) glycidyl monovinyl ester or glycidyl monovinyl ether and (B) polyvinyl ester of polyhydric alcohol, and the component (A) is (
Propane sultone or 1,4- Butane sultone is applied to the hydroxyl group to form the general formula (II) ▲There are mathematical formulas, chemical formulas, tables, etc.▼...(II) (wherein, X represents a propylene group or a butylene group,
l represents 0 or 1, m represents 0 or an integer from 1 to 15, n represents an integer from 2 to 4, and when l is 1, m is 0. 1. A method for producing a strongly acidic cation exchange resin, which comprises bonding atomic groups represented by ) to obtain a porous copolymer. (3) Main components are (A) glycidyl monovinyl ester or glycidyl monovinyl ether and (B) polyvinyl ester of polyhydric alcohol, and the component (A) is (
Epihalohydrin, ethylene glycol diglycidyl ether is added to the glycidyl group based on component (A) of the gel copolymer crosslinked with component B), or to the hydroxyl group generated by ring-opening modification of the glycidyl group with water or glycerin. , propylene glycol diglycidyl ether, 1,
3-propanediol diglycidyl ether, 1,4-
A sulfite compound is applied to the glycidyl group of a modified gel-like copolymer obtained by bonding butanediol diglycidyl ether or polyethylene glycol dicrycidyl ether, and the glycidyl group is converted into the general formula (III)▲mathematical formula, chemical formula, table etc. ▼ ...(III) (In the formula, l represents 0, 1 or 2, m represents 0 or 1
n represents an integer from 2 to 4, p represents 0 or 1, when p is 0, l and m are 0, and when l is 0, m is 0. , when l is 1, m is not 0. 1. A method for producing a strongly acidic cation exchange resin, which comprises bonding atomic groups represented by ) to obtain a porous copolymer.