JPS61142459A - Manufacture of filler for liquid chromatograph - Google Patents

Abstract

PURPOSE:To obtain a filler which provides a large strength and a high separation performance with an even mesh structure, by making polymer particles mainly composed of glycidyl monovinyl ester or glycidyl monovinyl ether hydrophylic by opening rings of epoxy groups after swelled in a proper solvent. CONSTITUTION:Glycidyl monovinyl ester or glycidyl monovinyl ether is used as main component and is polymerized in suspension with a proper amount of divinyl benzen or the like to introduce a desired crosslinked bond in addition to other copolymerizable monomers as required to obtain a polymer particle with the average particle size of 5-500mum. This particle is swelled in an organic solvent for swelling, such as aceton, and dioxane and made to react with polyvalent alcohol, amine or the like able to impart a hydrophylic group by opening the rings of epoxy groups to obtain a filler for a high performance liquid chromatography which is ideal for separation of a desired component with an excellent mechanical strength. This also can be utilized for industrial applications by massive filling thereof into a huge column.

Description

DETAILED DESCRIPTION OF THE INVENTION A. Field of Industrial Application The present invention relates to a method for producing a filler for liquid chroma 1-(LC).

In recent years, LC has been used not only as a means of analysis, but also in many industrial fields such as the food industry, pharmaceutical industry, synthesis and purification of chemical industry product intermediates, inorganic industry, and textile industry. As such gels for LC, dextran cross-linked polymer gels, acrylamide cross-linked polymer gels, cross-linked polystyrene gels, cross-linked polyvinyl acetate gels, and cross-linked polyethylene glycol dimethacrylate gels have been increasingly used as separation means. A wide variety of products have been developed, including silanol-based gels and silanol/polyoxyethylene-based gels. However, gels with high separation performance and high throughput are generally expensive; for example, Pharmacia Fine Chemical, which is known as a high-performance LC gel, is expensive.
Cef7Dex gel (dextran cross-linked polymer gel) manufactured by S Company and others has excellent performance in small-scale fractional FIA applications, but because it is a so-called soft gel, it has poor mechanical strength and cannot be used in large quantities in huge columns. In industrial applications where gel is used as a filler, its performance is not fully demonstrated due to destruction or compression of the gel, and it also causes operational problems such as increased operating pressure and increased frequency of backwash operations. Of course, this method has the inherent drawback of using a large amount of expensive gel.

As a result of intensive research to provide a bead-shaped polymer that is inexpensive and has high separation performance, the present inventors have found that by subjecting an epoxy group-containing spherical polymer to acidic heat treatment under specific conditions, the hydroxyl group can be removed by ring opening of the jepoxy group. was introduced and found that it was possible to provide an industrially advantageous LC gel with high resolution, and proposed Japanese Patent Application No. 108422/1983. Although such gels are inexpensive, have excellent separation performance, and have no defects in industrial operation, the ring-opening reaction proceeds from the particle surface, and the uniformity between the particle surface and the interior is not necessarily sufficient.

C. Problems to be Solved by the Invention The present invention solves the conventional drawbacks in the field of LC fillers, forms a uniform network structure from the particle surface to the inside, and introduces hydrophilic groups, and is inexpensive. It has high resolution and
Moreover, it is an object of the present invention to produce an LC filler that is free from defects in industrial operation and is industrially advantageous.

2. Means for Solving the Problems The above-mentioned object of the present invention is to swell polymer particles formed by polymerizing a monomer mainly composed of glycidyl monovinyl ester or glycidyl monovinyl ether with respect to the polymer. The present invention will be described in detail below, but first, the epoxy group-containing polymer used in the present invention is , glycidyl monovinyl ester or glycidyl monovinyl ether as the main component. In addition, glycidyl monovinyl ester or glycidyl monovinyl ester has a carbon number of 8.
-12 glycidyl esters of monovinyl carboxylic acids or glycidyl ethers of monovinyl alcohols having 8 to 12 carbon atoms, among which preferred examples include glycidyl acrylate, glycidyl methacrylate, allyl glycidyl ether, methallyl glycidyl ether, etc. I can do it.

There is no problem in using other monomers that can be copolymerized with the above-mentioned monomer as the main component, for example, vinyl chloride, vinyl A, vinyl halides such as vinyl chloride 7, etc. and halogenated vinyliso 7; unsaturated carboxylic acids such as acrylic acid, methacrylic acid, itaconic acid and their salts; esters of acrylic acid or methacrylic acid such as methyl, ethyl, butyl, octyl, methoxyethyl; methyl vinyl ketone , unsaturated ketones such as methyl 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 substituted products; vinyl sulfonic acid, methallyl sulfonic acid, Unsaturated hydrocarbon sulfonic acids such as p-styrene sulfone and salts thereof;
Styrene and its alkyl or halogen substituted products; allyl alcohol and hiso esters or ethers; vinyl compounds such as vinylidene cyanide and allyl methacrylate. In addition, as necessary, divinylbenzene, ethylene glycol dimethacrylate, diethylene glycol dimethacrylate, polyethylene glycol dimethacrylate), N,N'-methylenebisacrylamide, divinyl succinate, vinyl methacrylate,
By using a crosslinking monomer such as allyl methacrylate, some crosslinking bonds can be introduced into the polymer in advance.

By using such monomers and carrying out suspension polymerization in a medium that does not dissolve the resulting polymer, an epoxy group-containing spherical polymer can be obtained. As such a polymerization medium, use an organic solvent that does not adversely affect the polymerization reaction and dissolves the monomer polymer but precipitates the polymer, as long as it does not cause ring opening of the epoxy group in the polymerization system. However, water-based systems are preferred from the viewpoint of ease of handling and economic efficiency.

Such aqueous suspension polymerization essentially involves ethylenically unsaturated carboxylic acid such as acrylic acid, methacrylic acid, maleic acid, itaconic acid, etc. ! or a salt thereof and an ethylenically unsaturated sulfonic acid such as styrene sulfonic acid or sulfopropyl methacrylate or a salt thereof in the presence of a water-soluble polymer having a bond thereto, a conventional radical polymerization initiator, for example 2.2' -azobisisobutyronitrile, 2,2'-azobis(2,
4-dimethylvaleronitrile), 2,2'-azobis(
This can be advantageously carried out by using noazo compounds such as 2,4-dimethyl-4-methoxyvaleronitrile), peroxides such as benzoyl peroxide, and di-t-butyl peroxide. Such suitable polymerization means are described in further detail in Japanese Patent Application No. 104785/1985 filed by the present applicant.

Note that the particle size of the gel finally obtained is almost uniquely determined by the size of the epoxy group-containing polymer particles, so L
5 to 500 μ, preferably 8, suitable for gel use for C
It is desirable to produce polymer particles having an average particle size within the range of 0 to 300 microns.

Next, the swelling process of the polymer particles, which plays an extremely important role in achieving the purpose of the present invention, will be described. As for the type of organic solvent, any organic solvent can be used as long as it has the ability to swell the polymer particles. For example, Ketones such as acetone and methyl ethyl ketone; ethers such as dioxane, ethylene glycol dimethyl ether, and diethylene glycol dimethyl ether; cellosolves such as methyl cellosolve and ethyl cellosolve; dimethyl formamide, dimethyl sulfoxide, ethylene carbonate, methoxyethylene oxyethyl butylene 1,8 points Examples include methyl ethyl ketone, dioxane ethylene glycol dimethyl ether,
Diethylene glycol dimethyl ether, ethylene carbonate, methoxyethylene oxyethyl polyethylene 1
, 3 borate can give favorable results in terms of separation ability, workability, toxicity, etc. of the final gel.

The amount of solvent used, swelling conditions such as temperature and time are determined as appropriate depending on the properties such as hardness, crosslinking density, and gel moisture content that should be imparted to the final gel. Although it is difficult to specify the amount of solvent to be used, the amount of solvent to be used should be at least 3 times the weight of the polymer particles, preferably 5 to 20 times the weight of the polymer particles.
Double the temperature, increase the temperature to 50~140'C, and increase the time to 0.5~140'C.
It is desirable to set it within a range of 5 hours.

Next, as a means for ring-opening and making hydrophilic the epoxy group having a binder in the swollen polymer particles, use any method that can ring-open the epoxy group and introduce a desired amount of cross-linking and hydrophilic groups. It is possible to do so, and such means are widespread.

Although it is difficult to list all of them due to (2) addition of acidic sulfite or sulfite or salts thereof; Depending on the field of application of the final product, LC filler, methods ■ to ■ and ■ may be used if a nonionic hydrophilic group is required to be introduced; Methods 1 and 2 can be selected and used as appropriate; however, method 2 is not preferred when a gel with a high gel moisture content is required.

In addition, for the above methods 1 to 2, acids such as formic acid, acetic acid, hydroxyacetic acid, sulfuric acid, nitric acid, hydrochloric acid, and perchloric acid; alkalis such as sulfur hydroxide, potassium hydroxide, and ammonia; Zinc fluoride, boron trifluoride ether complex salt,
By reacting zinc chloride etc. for about 0.5 to 5 hours, preferably at a humidity temperature of 0'C or higher, it is industrially advantageous to open the epoxy group, form a crosslink, and improve hydrophilic properties. It is possible to introduce a group (hydroxyl group). In addition, the polyhydric alcohol used in the above method (2) includes glycols such as ethylene glycol, pro-1,2,8-glycol, trimethylene glycol, tetramethylene glycol, polyethylene glycol, and polypropylene glycol, glycerin, 1,2,8- Octahydric alcohols such as phythanetriol; Tetrahydric alcohols such as erythritol, pentaerythritol, and D-xylose; Pentavalent or higher alcohols such as D-arabitol, D-mannitol, D-turhite, manonite, etc. You can list the following,
Among them, glycols and trihydric alcohols are preferred.

In addition, the above method (2) includes tertiary amines such as benzyldimethylamine, triphylylamino, and tris(dimethylamino)methylphenol; quaternary ammonium salts such as triethylbenzylammonium chloride and tetramethylammonium chloride; 2-methylimidazole;
HyFO-IP cyacetic acid, lactic acid,
Oxymonocarboxylic acids such as α, β-dioxyisobutyric acid, trioxybutyric acid, and gluconic acid; Monooxypolycarboxylic acids such as malic acid, oxyglutaric acid, and citric acid; Tartaric acid, α, β-dioxyglutaric acid, and oxycitric acid ,
Hydroxycarboxylic acids such as polyoxypolycarboxylic acids such as trioxybutyric acid can be added.

Furthermore, as the above method (2), ammonia, hydroxylamine, hydrazine, monoethanolamine, jetanolamine, triethanolamine, 2-oxy-2-
Amines such as methylpropylamine, urea, 1,000 urea, 29-thousand urea, etc., and methyl mercaptan, monothioethylene glycol, thioglycol, ethanedithio-/', 1,4-butanedithiol, etc. These mercaptans can be easily added without using a catalyst. In addition, in any of the above-mentioned methods (1) to (2), the same temperature and time conditions as in the above-mentioned methods (1) to (2) can be employed.

Note that when water is used as a medium for the ring-opening of epoxy groups, cross-linking, and introduction of hydrophilic groups, the hydrolysis reaction proceeds preferentially and In order to prevent the introduction of functional groups other than those mentioned above, or from an industrial point of view, the organic solvent used in the swelling step is continued to be used, and the reaction is carried out in a homogeneous system or, in some cases, in a cine homogeneous system. It is preferable that water is used in combination, and the amount of water should be kept to the minimum necessary.

In this way, particle sizes generally range from 10 to 500, u
, preferably 80-3ooμ, gel moisture content 30-20
00%, preferably 50 to 1000% gel can be manufactured with industrial advantage.

2. Function By adopting the above-mentioned means of the present invention, the epoxy group-containing polymer particles are swollen, and uniform cross-linking is formed over the surface and inside of the particles, and hydrophilic groups are introduced and the particles are swollen. It is believed that the state particle structure is fixed, and the present invention provides a gel with a high crosslinking density and a high gel water content, that is, a packing material for LC that has good mechanical strength and separation ability. It is considered possible.

E. Effects of the invention: The crosslinking density and gel moisture content are high, and the particle structure is uniform.Therefore, there are no operational defects due to increased operating pressure or increased frequency of backwashing operations, and high separation is achieved. LC with the ability
A characteristic advantage of the present invention is that it can provide a filler for epoxy groups. Another advantage of the present invention is that the amount of cross-linking introduced can be arbitrarily controlled without adjusting the amount of cross-linkable monomer at the stage of preparing the group-containing polymer and preparing a wide variety of polymers. It is.

The gel of the present invention produced in this way is inexpensive, has high efficiency, and has no operational problems, and can be used for gel filtration using the molecular sieve effect, or for ion chromatography by introducing ionic groups. It is used industrially in an extremely wide variety of fields of application, including as a packing material for adsorption chromatography, and as a packing material for adsorption chromatography.

EXAMPLES The present invention will be explained in more detail with reference to Examples below, but the scope of the present invention is not limited in any way by the description of these Examples. In the examples, parts and percentages are expressed on a weight basis unless otherwise specified.The gel moisture content is calculated by placing polymer particles sufficiently equilibrated with deionized water in a centrifugal separator with a centrifugal effect of 2000G to adhere to the particle surface. After removing the water present, the weight (Wl) of the polymer particles is measured, and then the polymer particles are dried so that the dry weight<Wz
> was measured and calculated using the following formula.

Example 1 2s parts of polymer particles with an average particle diameter of 180μ obtained by main polymerization of glycidyl methacrylate (GMA) and 175 parts of ethylene carbonate (EC) were charged into a flask, heated to 120C, and stirred for 1 hour. After that, 60 parts of glycerin was added, and after further stirring for 1 hour, 0 parts of zinc borofluoride was added.
．． 2 parts were added and reacted for 8 hours.

The gel moisture content of the obtained gel particles (a) was 278%.

This gel was packed into a glass column with an inner diameter of 1.5 cN and a height of 80 cIIt, and its performance as a packing material for LC was evaluated. Bovine serum albumin 596 and ammonium sulfate 2.
A column was charged with 0.5 cc of an aqueous solution containing 5%, deionized water was used as an eluent, and the elution rate was 1 cc for 7 minutes to create a dissolution curve as shown in FIG. 1(a).

Table 1 also shows the results of measuring the relationship between the elution rate and pressure drop of this column.

For comparison, a gel (b, gel water 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 Figure 1 ( b)
and listed in Table 1.

From the results shown in Table 1, Figure 1, and Table 1, it can be understood that the product of the present invention has excellent sweat resistance and degreasing performance.

Example 2 25 parts of the polymer particles of Example 1 and 175 parts of methyl ethyl ketone were placed in a flask and stirred at 50''C for 2 hours, then 70 parts of jetanolamine was added and reacted with stirring for 8 hours.

After the reaction, gel particles (e) were washed with deionized water, and gel particles (d) were neutralized with 10% hydrochloric acid 101 after the reaction and washed with deionized water. The gel moisture content was measured to be 247%. , 578%.

Using gel particles (d), Amy was prepared in the same manner as in Example 1.
The results of electrical distribution of cal No5L (trade name of polysaccharide mixture manufactured by Nisso Kagaku ■) are shown in FIG.

Example 3 Diethylene glycol dimethyl ether as a substitute for EC,
The same procedure as in Example 1 was carried out except that 70 parts of gluconic acid was used instead of 60 parts of glycerin, 0.5 part of triethylbenzylammonium chloride was used instead of 0.1 part of perchloric acid, and the reaction temperature was adjusted to t-130'C. Gel particles (e) were prepared using the gel particles. The gel moisture content of these gel particles was 28,896. The separation ability of Amycol N (L 6 L) was evaluated in the same manner as in Example 2. The results are shown in FIG.

Example 4 25 parts of polymer particles with an average particle diameter of 120 μ obtained by polymerizing GMA9596 and 5% vinyl acetate and 175 parts of the swelling ff4 agent shown in Table 2 below were charged into a flask and heated at 90″C.
After stirring for 1 hour, 70 parts of deionized water was added, and after further stirring for 1 hour, 0.1 part of perchloric acid was added and reacted for 8 hours. After the reaction was completed, four types of gel particles (fi) were obtained by washing with deionized water.

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 results of measuring the gel moisture content are also listed in Table 2. do.

Table 2 Also, the results of evaluating the separation ability of the gel particles (f and j) according to the formulation of Example 1 are shown in FIG. 4, and it can be seen that the product of the present invention has excellent separation ability.

[Brief explanation of the drawing]

Figure 1 is from Example 1 (Pk & and b), Figure 2 is from Example 2 (NLd), and Figure 8 is from Example 8 (No, e).
), and FIG. 4 shows Example 4 (No, f and j).
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noon o 5゜jibanan) ((Kogen0 10 '20 30 4()
Fox v 505! jr-'jf, ($劫妾-?Yama Ichibanzut*+ (ml)

Claims (1)

[Claims] 1. Polymer particles obtained by polymerizing a monomer containing glycidyl monovinyl ester or glycidyl monovinyl ether as a main component are swollen in an organic solvent that has a swelling ability for the polymer. A method for producing a packing material for liquid chromatography, which comprises subsequently ring-opening the epoxy group and making it hydrophilic. 2. The manufacturing method according to claim 1, wherein a polyhydric alcohol is added to an organic solvent.