JPH0750087B2 - Method for manufacturing packing material for liquid chromatography - Google Patents

Description

TECHNICAL FIELD The present invention relates to a method for producing a packing material for liquid chromatography (LC).

(B) Conventional technology LC has been used not only as one of analytical means in recent years, but also in many fields such as food industry, pharmaceutical industry, synthesis and purification of intermediate products for chemical industry, inorganic industry, textile industry, etc. Is being applied as one of the effective separation means, and such LC
As gel for gel, dextran cross-linked polymer gel, acrylamide cross-linked polymer gel, cross-linked polystyrene gel, cross-linked polyvinyl acetate gel, cross-linked polystyrene glycol dimethacrylate gel, silanol polyoxyethylene gel, etc. have been developed. There is. However, gels with higher separation performance and higher processing capacity are generally more expensive. For example, Ph that is known as a high performance LC gel
Sephadex gels (dextran cross-linked polymer gels) from armacia Fine chemicals, etc. have excellent performance in small-scale separation applications, but they are so-called soft gels and have poor mechanical strength. In industrial applications where the gel is used for filling, not only its performance is not fully exhibited due to breakage and compression of gel, but it also causes operational problems such as increase in operating pressure and increase in backwashing operation frequency. Of course, it has an inherent defect that a large amount of expensive gel is used.

Therefore, the present inventors have conducted intensive studies to provide a bead-shaped polymer that is inexpensive and has high separation performance, and as a result, the epoxy group-containing spherical polymer is subjected to acidic heat treatment under specific conditions to open the epoxy group to form a hydroxyl group. Was introduced, and the fact that a gel for LC having high resolution can be provided industrially advantageously was proposed, and the Japanese Patent Application No. 58-108422 was previously proposed. Although such a gel is inexpensive and has excellent separation performance and has no defects in industrial operation, the ring-opening reaction proceeds from the particle surface, and the uniformity between the particle surface and the inside is not always sufficient.

(C) Problems to be Solved by the Invention The present invention eliminates the conventional drawbacks in the field of LC fillers, forms a uniform network structure from the particle surface to the inside, and introduces a hydrophilic group, which is inexpensive. With high resolution,
Moreover, it is an object of the present invention to industrially produce a filler for LC that has no defects in industrial operation.

D, means for solving the problems The above-described object of the present invention is to provide a polymer particle comprising a glycidyl monovinyl ester or a glycidyl monovinyl ether as a main component and a monomer containing no crosslinkable monomer. This is achieved by swelling the polymer in an organic solvent having swelling ability, and then ring-opening and hydrophilizing the epoxy group.

Hereinafter, the present invention will be described in detail. First, the epoxy group-containing polymer used in the present invention is produced by using glycidyl monovinyl ester or glycidyl monovinyl ether as a main component. The glycidyl monovinyl ester or glycidyl monovinyl ether has 3 carbon atoms.
To 12 monovinylcarboxylic acid glycidyl esters or C3 to C12 monovinyl alcohol glycidyl ethers, among which glycidyl acrylate, glycidyl methacrylate, allyl glycidyl ether, and methallyl glycidyl ether are preferred examples. You can

It should be noted that the above-mentioned monomer which is the main component may be appropriately used in combination with another monomer which can be copolymerized with the monomer, and for example, vinyl chloride, vinyl bromide, vinyl halide such as vinylidene chloride and vinyl halide and Vinylidene halides; unsaturated carboxylic acids such as acrylic acid, methacrylic acid, itaconic acid and salts thereof; esters of acrylic acid or methacrylic acid such as methyl, ethyl, butyl, octyl and methoxyethyl; methyl vinyl ketone, methyl Unsaturated ketones such as isopropenyl ketone; vinyl esters such as vinyl formate, vinyl acetate and vinyl propionate; vinyl ethers such as methyl vinyl ether and ethyl vinyl ether; acrylamide and its alkyl substitution products; vinyl sulfonic acid, methallyl sulfonic acid Carbonization of p-styrene sulfonic acid, etc. Containing sulfonic acid and salts thereof; styrene and its alkyl or halogen substituents;
Examples thereof include allyl alcohol and its esters or ethers; vinyl compounds such as vinylidene cyanide and methacrylonitrile.

The epoxy group-containing spherical polymer can be obtained by carrying out suspension polymerization using a monomer as described above in a medium that does not dissolve the resulting polymer. As such a polymerization medium, it is possible to use an organic solvent which does not adversely affect the polymerization reaction and dissolves the monomer mixture but precipitates the polymer unless it causes the ring opening of the epoxy group in the polymerization system. However, an aqueous system is preferably used from the viewpoint of easy handling and economy.

Such aqueous suspension polymerization is substantially carried out by using an ethylenically unsaturated carboxylic acid such as acrylic acid, methacrylic acid, maleic acid or itaconic acid or a salt thereof and an ethylenically unsaturated sulfonic acid such as styrene sulfonic acid or sulfopropyl methacrylate. Or in the presence of a water-soluble polymer containing a bond with a salt thereof, an ordinary radical polymerization initiator such as 2,2′-azobisisobutyronitrile or 2,2′-azobis (2,4-dimethylvalero) is used. Nitrile), 2,2'-azobis (2,4-dimethyl-4-methoxyvaleronitrile) and other azo compounds, and benzoyl peroxide, di-t-butyl peroxide, and other peroxides are advantageous. Can be done.
The suitable polymerization means is described in more detail in Japanese Patent Application No. 58-104735 assigned by the present applicant.

Since the particle size of the finally obtained gel is almost uniquely determined by the size of the epoxy group-containing polymer particles, LC
5 to 500μ, preferably 30 to 300, suitable for gel use
It is desirable to make polymer particles having an average particle size in the range of μ.

Next, the swelling step of the polymer particles, which plays an extremely important role in achieving the object of the present invention, will be described.

As the type of the organic solvent, any organic solvent can be used as long as it has a swelling ability with respect to the polymer particles, and examples thereof include ketones such as acetone and methyl ethyl ketone; ethers such as dioxane, ethylene glycol dimethyl ether and diethylene glycol dimethyl ether; methyl cellosolve and ethyl. Cellosolves such as cellosolve; dimethylformamide, dimethylsulfoxide, ethylene carbonate, methoxyethyleneoxyethylbutylene 1,3 borate and the like can be mentioned, but among them, methyl ethyl ketone,
If dioxane ethylene glycol dimethyl ether, diethylene glycol dimethyl ether, ethylene carbonate, methoxyethylene oxyethyl butylene 1,3 borate is used, the resolution of the gel finally obtained,
It is possible to give favorable results in terms of workability and toxicity.

The amount of the solvent used, the swelling conditions such as temperature and time, are appropriately determined according to various properties such as hardness, cross-linking density and gel moisture content that should be imparted to the finally obtained gel. However, the amount of the solvent used is 3 times or more, preferably 5 to 20 times the weight of the polymer particles.
It is desirable to set the temperature to 50 to 140 ° C. and the time to 0.5 to 5 hours.

Next, as a means for ring-opening and hydrophilizing the epoxy group contained in the swollen polymer particles, it is preferable to use as long as the epoxy group can be ring-opened and a desired amount of cross-linking and hydrophilic group can be introduced. However, it is difficult to list all of them because such means are extremely diverse, but typical examples thereof include hydrolysis, addition of polyhydric alcohol, addition of alkyl alcohol, addition of hydroxycarboxylic acid, and addition of amine. Addition, addition of mercaptan, addition of acidic sulfurous acid or sulfurous acid or salts thereof, and the like can be mentioned. Incidentally, depending on the field of application of the final product LC filler, when the introduction of a nonionic hydrophilic group is required ~ and method, from the purpose of introducing an ionic hydrophilic group, and the method. Can be appropriately selected and used, but the method is not preferable when a gel having a high gel water content is required.

Regarding the above-mentioned methods, as catalysts, acids such as formic acid, acetic acid, hydroxyacetic acid, sulfuric acid, nitric acid, hydrochloric acid, perchloric acid; alkalis such as sodium hydroxide, potassium hydroxide, ammonium, zinc borofluoride, trifluoride, etc. Boron ether complex salt,
By using zinc chloride or the like, preferably at a temperature of 50 ° C. or higher, by reacting for approximately 0.5 to 5 hours, industrially advantageous ring opening of epoxy groups, formation of cross-linking, and formation of hydrophilic groups (hydroxyl groups) Can be introduced. Examples of the polyhydric alcohol used in the above method include glycols such as ethylene glycol, propylene glycol, triethylene glycol, tetramethylene glycol, polyethylene glycol and polypropylene glycol; glycerin, 1,2,3-butanetriol and the like 3 Polyhydric alcohols; tetrahydric alcohols such as erythritol, pentaerythritol and D-xylose; pentavalent or higher alcohols such as D-arabite, D-mannite, D-sorbit and mannite, Of these, glycols and trihydric alcohols are preferable.

Further, as the above method, tertiary amines such as benzyldimethylamine, tributylamine and tris (dimethylamino) methylphenol; quaternary ammonium salts such as triethylbenzylammonium chloride and tetramethylammonium chloride; 2-methylimidazole, 2-
In the presence of a catalyst such as imidazoles such as methyl-4-ethylimidazole, hydroxyacetic acid, lactic acid, α, β-
Oxymonocarboxylic acids such as dioxyisobutyric acid, trioxybutyric acid and gluconic acid; malic acid, oxyglutaric acid,
Monooxypolycarboxylic acids such as citric acid; tartaric acid, α,
Hydroxycarboxylic acids such as polyoxypolycarboxylic acids such as β-dioxyglutaric acid, oxycitric acid and trioxybutyric acid can be added.

Further, as the above method, ammonia, hydroxylamine, hydrazine, monoethanolamine, diethanolamine, triethanolamine, 2-oxy-2-
Amines such as methylpropylamine, urea, thiourea, and thiourea dioxide, and as the above method, mercaptans such as methyl mercaptan, monothioethylene glycol, thioglycolic acid, ethanedithiol, and 1,4-butanedithiol, It can be easily added without using a catalyst. In any of the above-mentioned methods,
The conditions of temperature and time substantially similar to the above-mentioned methods can be adopted.

When water is used as a medium for carrying out the ring-opening of the epoxy group, the crosslinking bond and the introduction reaction of the hydrophilic group, the hydrolysis reaction preferentially proceeds to form the crosslinking bond or the addition reaction (hydroxyl group). It is difficult to introduce functional groups other than), or from the industrial viewpoint, the organic solvent used in the swelling step is continuously used to carry out the reaction in a homogeneous system or in a heterogeneous system in some cases. However, when water is used in combination, the amount of water should be kept to the minimum necessary.

In this way, generally particle size is 10 ~ 500μ, preferably 30 ~ 300μ, gel moisture content 30 ~ 2000%, preferably 50.
~ 1000% gel can be produced industrially advantageously.

D. By adopting the above-described means of the present invention, the epoxy group-containing polymer particles are swollen, the formation of uniform cross-linking and the introduction of hydrophilic groups and immobilization of the swollen state particle structure over the surface and inside of the particles. Therefore, it is considered that a gel having a high crosslink density and a high gel water content, that is, an LC melt filler having excellent mechanical strength and excellent separability can be provided.

E. Effect of the invention The crosslink density and gel moisture content are high, and the particle structure is uniform, so there is no operational defect due to increase in operating pressure or frequency of backwashing operation, and high separation. It is a characteristic advantage of the present invention that a filler for LC having a function can be provided.

In addition, by adjusting the amount of polyhydric alcohol, hydroxycarboxylic acid, amine, etc. added to the epoxy group, the amount of the crosslinkable monomer can be adjusted at the stage of preparing the epoxy group-containing polymer in advance to prepare various types. It is also an effect of the present invention that the amount of cross-linking introduced can be arbitrarily controlled without preparing a polymer extending over the entire length.

The gel of the present invention produced in this manner is inexpensive, has high resolution, and has no operational problems. Therefore, it can be used for gel chromatography utilizing the molecular sieving effect or for ion chromatography by introducing an ionic group. As a packing material for adsorption chromatography, it is industrially used in a wide variety of application fields.

F. Examples Hereinafter, the present invention will be described in more detail with reference to Examples, but the scope of the present invention is not limited by the description of these Examples. In the examples, parts and percentages are by weight unless otherwise specified. The gel water content was determined by subjecting polymer particles sufficiently equilibrated with deionized water to a 2000 G centrifugal separator to remove water adhering to the particle surface, and then measuring the weight (W ₁ ) of the water. The polymer particles were dried, and the weight (W ₂ ) after drying was measured, and the value was obtained by the following formula.

Example 1 25 parts of polymer particles having an average particle diameter of 180μ obtained by polymerizing glycidyl methacrylate (GMA) and 175 parts of ethylene carbonate (EC) were charged in a flask, heated to 120 ° C. and stirred for 1 hour. Then, 60 parts of glycerin was added, and the mixture was further stirred for 1 hour, then 0.2 part of zinc borofluoride was added and reacted for 3 hours.

The gel moisture content of the obtained gel particles (a) was 273%.

This gel was packed in a glass column having an inner diameter of 1.5 cm and a height of 30 cm to evaluate the performance as a packing material for LC. Aqueous solution containing 5% bovine serum albumin and 2.5% ammonium sulfate as a sample
0.5 cc was charged in the column, deionized water was used as an eluent, and the elution rate was 1 cc / min, and the elution curve shown in FIG. 1 (a) was prepared.

Table 1 shows the results of measurement of the relationship between the elution rate of this column and the pressure loss.

For comparison, a gel (b, gel moisture content: 90%) was prepared in the same manner as above except that deionized water was used instead of EC, and the results evaluated in the same manner as above are shown in FIG. ) And Table 1.

From the results of FIG. 1 and Table 1, it is understood that the product of the present invention has excellent pressure resistance and separation performance.

Example 2 25 parts of the polymer element described in Example 1 and 17 of methyl ethyl ketone
After charging 5 parts to a flask and stirring at 50 ° C. for 2 hours, 70 parts of diethanolamine was added and the reaction was carried out while stirring for 3 hours.

After the reaction, gel water content was measured for gel particles (c) washed with deionized water and gel particles (d) washed with deionized water after neutralization with 10% hydrochloric acid 10 after the reaction.
It was 7% and 573%.

Using the gel particles (d) and following the same procedure as in Example 1, Amycol
FIG. 2 shows the result of separation of No. 6L (trade name of polysaccharide mixture manufactured by Nitto Chemical Co., Ltd.).

Example 3 Diethylene glycol dimethyl ether in place of EC, 70 parts of glycolic acid in place of 60 parts of glycerin, triethylbenzylammonium chloride in place of 0.1 part of perchloric acid.
Gel particles (e) were prepared in the same manner as in Example 1 except that 5 parts were used and the reaction temperature was 130 ° C.

The gel moisture content of this gel particle was 238%, and the results of evaluating the separation ability of Amycol No. 6L as in Example 2 are shown in FIG.

Example 4 25 parts of polymer particles having an average particle diameter of 120μ obtained by polymerizing 95% of GMA and 5% of vinyl acetate and a swelling agent 1 shown in Table 2 below.
75 parts was placed in a flask and stirred at 90 ° C. for 1 hour, 70 parts of deionized water was added, and after stirring for another 1 hour, 0.1 part of perchloric acid was added and reacted for 3 hours. After completion of the reaction, the gel was washed with deionized water to obtain four types of gel particles (fi).

The results of measuring the water content of these gels are shown in Table 2 below. For comparison, gel particles (j) were prepared in the same manner as above except that deionized water was used instead of the swelling agent, and the gel moisture content was measured.

Further, the results of evaluation of the separability of gel particles (f and j) according to the formulation of Example 1 are shown in FIG. 4, and it is understood that the product of the present invention has excellent separability.

[Brief description of drawings]

FIG. 1 is from Example 1 (No.a and b), FIG. 2 is from Example 2 (No.d), FIG. 3 is from Example 3 (No.e), and FIG. The elution curves obtained from Example 4 (Nos. F and j) are shown.

Claims (2)

[Claims] 1. An organic solvent having swelling ability for polymer particles, which comprises glycidyl monovinyl ester or glycidyl monovinyl ether as a main component and is obtained by polymerizing a monomer containing no crosslinkable monomer. A method for producing a packing material for liquid chromatography, which comprises swelling in an epoxy group, and then ring-opening and hydrophilizing an epoxy group. 2. The method according to claim 1, wherein a polyhydric alcohol is added to the organic medium.