JPH0236351A - Production of column packing material for liquid chromatography - Google Patents

Abstract

PURPOSE:To improve acid resistance and alkali resistance by hydrolyzing the gel particles obtd. by bringing glycidyl monovinyl ester and polyvinyl ester of polyhydric alcohol into aq. suspension copolymn. in the presence of an acid which does not react with the glycidyl group of said particles and bringing 6-24C fatty acid into reaction. CONSTITUTION:Preferably glycidyl methacrylate as the glycidyl monovinyl ester, etc., and, for example, di and tri of ethylene glycol dimethacrylate as the polyvinyl ester of polyhydric alcohol, etc., are brought into the aq. suspension copolymn. The glycidyl group of the resulted gel particles is then hydrolyzed in the presence of the acid which does not react with the glycidyl group. Further, 6-24C fatty acid, fatty acid anhydride or fatty acid chloride are brought into reaction. The acid resistance and alkali resistance are improved in such a manner.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a method for producing a column packing material for liquid chromatography, particularly a column packing material for reverse phase partition liquid chromatography.

[Conventional technology]

Conventionally, as one separation method of liquid chromatography,
Reversed phase partition chromatography is known, which separates samples based on the strength of retention due to differences in hydrophobicity. There is.

(1) Chemically bonded silica gel chemically modified with a silane coupling agent having octadecyl groups etc. on the surface of silica gel [G.A. Kirkland, J.J.
D'Esteefano, Journal of Chromatographic Science
5science) Volume 8, page 309,
1970] (2) Styrene-divinylbenzene copolymer particles [M.D. Greaser, D.G. Beatsack, Analytical Chemistry (Analytica)
l Chemistry) Volume 45, Page 1383, 19
(1973) (3) Methacrylic acid ester copolymer particles having long alkyl chains (JP-A-62-90533) [Problems to be solved by the invention] Of the above, the silica gel in (1) is alkali-based. It has the disadvantage that the pn of the eluent is limited to 2 to 8 and cannot be used in all pH ranges.Furthermore, it is difficult to chemically bond to all the silanol groups on the silica surface, and unreacted silanol Since the groups remain, there is a problem that specific adsorption occurs for basic samples.

In addition, although the copolymer particles (2) and (3) have solved the problem regarding silica gel in (1), they swell and shrink rapidly between various solvents, so it is necessary to change the eluent in the column. There is a problem in that it is difficult to perform chromatography.

An object of the present invention is to solve these problems and provide a column packing material for reverse phase partition liquid chromatography that has acid resistance and alkali resistance, and has small differences in swelling and shrinkage between various solvents. .

[Means for Solving the Problems] The present invention provides glycidyl groups in gel particles obtained by aqueous suspension copolymerization of glycidyl monovinyl ester or glycidyl monovinyl ether and polyvinyl ester of polyhydric alcohol or polyvinyl ether of polyhydric alcohol. is hydrolyzed in the presence of an acid that does not react with the glycidyl group, and further hydrolyzed with a fatty acid having 6 to 24 carbon atoms, and a fatty acid having 6 to 24 carbon atoms.
The present invention relates to a method for producing a column packing material for liquid chromatography, which is characterized by reacting a fatty acid anhydride having 24 carbon atoms or a fatty acid chloride having 6 to 24 carbon atoms.

Examples of the glycidyl monovinyl ester or glycidyl monovinyl ether used in the present invention include glycidyl methacrylate, glycidyl acrylate, and allyl glycidyl ether, but glycidyl methacrylate is preferred in consideration of reactivity and alkali resistance.

The polyvinyl ester of polyhydric alcohol and the polyvinyl ether of polyhydric alcohol used in the present invention include:
For example, alkylene glycol divinyl esters such as ethylene glycol dimethacrylate, ethylene glycol diacrylate, propylene glycol dimethacrylate, and propylene glycol diacrylate; divinyl ester of alkylene glycol,
Di- or triacrylate of glycerin, di- or trimethacrylate of glycerin, di- or trimethacrylate of trimethylolpropane, di- or triacrylate of trimethylolpropane, di- or trimethylolmethane,
tri- or tetramethacrylate, di-, tri- or tetraacrylate of tetramethylolmethane, ethylene glycol diallyl ether, propylene glycol diallyl ether, polyethylene glycol diallyl ether,
Examples include polypropylene glycol diallyl ether, di- or triallyl ether of glycerin, di- or triallyl ether of trimethylolpropane, di-, tri- or tetraallyl ether of tetramethylolmethane, etc.; In consideration of pressure resistance, ethylene glycol dimethacrylate and tetramethylolmethane di, tri, or tetramethacrylate are desirable.

Glycidyl monovinyl ester or glycidyl monovinyl ether (I) and polyvinyl ester of polyhydric alcohol or polyvinyl ether of polyhydric alcohol (II)
If (1) is too large, the hydrophilicity of the generated gel particles will improve, but the pressure resistance will tend to decrease;
The ratio of 1)/(n) is 50150 to 97/ in molar ratio.
A range of 3 is preferred.

In the present invention, the glycidyl monovinyl ester or glycidyl monovinyl ether and the polyvinyl ester of a polyhydric alcohol or the polyvinyl ether of a polyhydric alcohol are subjected to aqueous suspension polymerization in the presence of an organic solvent that is incompatible with water, if necessary, to form a gel. It is considered to be a particle.

The water-incompatible organic solvents used in the present invention are used to make the gel porous and these
The amount dissolved in 0 g is 15 g or less, such as toluene, diethylbenzene, dodecane, isoamyl alcohol, chlorobenzene, ethyl acetate, propyl acetate, butyl acetate, and the like. These solvents may be used alone or in combination of two or more. The amount of these used is preferably 50 to 300% by weight based on the total amount of monomers. If there is too little organic solvent that is incompatible with water, it will be difficult to make the resulting particles porous.
If the amount is too large, the porosity of the resulting particles tends to increase, resulting in poor pressure resistance.

Aqueous suspension polymerization involves suspension polymerization in an aqueous medium, and water is essential as this aqueous medium, and a water-soluble organic solvent must be dissolved within the range that does not impair the stability of the suspension system. You may use water that has been drained.

At this time, it is preferable to add all the organic solvents that are incompatible with water before the reaction or until the polymerization rate reaches 20%. The addition may be done in portions.

Polymerization initiators suitable for use in the present invention include benzoyl peroxide, dichlorobenzoyl peroxide, dicumyl peroxide, di-tert-butylene peroxide 7L/2. 5-cy(peroxybenzoate)hexyne-3,1,3-bis(tert-butylperoxyisopropyl)benzene,
Lauroyl peroxide, tert-butyl peracetate, 2,5-dimethyl-2,5-di(tert-butylperoxy)hexyne-3
, 2゜5-dimethyl-2,5-di(tert-butylperoxy)hexane and tert-butyl perbenzoate, organic peroxides such as methyl ethyl ketone peroxide, methyl cyclohexanone peroxide, azobisisobutyronitrile and dimethyl azo There are azo compounds such as diisobutyrate, and one or more of these can be used. The amount used is determined depending on the type of vinyl monomer and the desired molecular weight of the resulting polymer, but it is preferably used in an amount of 0.1 to 4.0% by weight based on the vinyl monomer. be done.

In addition, in the present invention, a poorly soluble phosphate as a dispersant,
Water-soluble polymeric protective colloids and the like can be added to the polymerization system.

Examples of sparingly soluble phosphates include tricalcium phosphate and magnesium phosphate. As a polymeric protective colloid,
Examples include polyvinyl alcohol, water-soluble cellulose derivatives such as alkylcellulose, hydroxyalkylcellulose, and carboxyalkylcellulose, and sodium polyacrylate. It is preferable that the sparingly soluble phosphate is used in an amount of 0.01% by weight or more, and the water-soluble polymer protective colloid is used in an amount of 1 to 0.001% by weight based on the total amount of substances present in the polymerization system.

In addition, anionic surfactants are added to the polymerization system for particle size control, etc., and water-soluble inorganic Salts can be added to the polymerization system.

Glycidyl monovinyl ester or glycidyl monovinyl ether, an organic solvent incompatible with water, a dispersant, a polymerization initiator, and, if necessary, a dispersion aid, are mixed in advance or each added separately to an aqueous medium and dispersed. . In this case, in order to disperse well, it is preferable to perform high-speed stirring using a homomixer or the like, and this high-speed stirring can be continued until the early stage of polymerization. The subsequent polymerization is carried out under normal stirring using a propeller stirrer or the like.

The aqueous medium is preferably used in an amount of 1 to 50 times the total amount of the glycidyl monovinyl ester or glycidyl monovinyl ether and the water-incompatible organic solvent. In this case, water is used as the aqueous medium, but water containing a water-soluble organic solvent dissolved therein may also be used as long as it does not impede the stability of the suspension system.

For hydrolyzing the glycidyl groups of the gel particles, an acid that does not react with the glycidyl groups is used. Such acids include sulfuric acid, perchloric acid, toluenesulfonic acid, benzenesulfonic acid, and the like. The initial concentration of the acid solution used is 0.1N ~
5N is preferred. If it is thinner than this, the reaction will not proceed, and if it is thicker than this, hydrolysis of the ester bond will tend to occur. Further, it is preferable that the acid solution contains at least 10% by weight of water.

The solvent used for preparing the above acid solution is not particularly limited as long as it is compatible with acid and water and is inert to acid and glycidyl groups, such as acetone, methyl ethyl ketone, dioxane, tetrahydrofuran, acetonitrile, etc. However, there is no problem in simply using water as a solvent without using these solvents.

The amount of acid solution used should be sufficient to immerse the gel particles in the solvent, and should be at least the same amount as the weight of the gel. This reaction is carried out at 25°C to 100°C for 3 to 10 hours. It will be done. As a result, 8 of the glycidyl group possessed by the gel particles
More than 0% is hydrolyzed.

When one glycidyl group is hydrolyzed by the action of an acid in this manner, one glycidyl group is generated.

A fatty acid having 6 to 24 carbon atoms, which is reacted with a gel obtained by hydrolyzing a glycidyl group in the presence of an acid, and a fatty acid having 6 to 24 carbon atoms.
There is no particular restriction on the fatty acid anhydride having 24 carbon atoms and the fatty acid chloride having 6 to 24 carbon atoms, but stearic acid chloride and lauric acid chloride are preferred in view of their industrial availability and high reactivity. .

Fatty acids having 6 to 24 carbon atoms, fatty acid anhydrides having 6 to 24 carbon atoms, fatty acid chlorides having 6 to 24 carbon atoms,
Glycidyl groups are hydrolyzed in the presence of acid (preferably at least 10-4 mol per gram of dry gel is used. If less than 10-4 mol per gram is used, the resulting gel is subjected to liquid chromatography. The performance as a column packing material becomes insufficient.

Fatty acids having 6 to 24 carbon atoms, fatty acid anhydrides having 6 to 24 carbon atoms, fatty acid chlorides having 6 to 24 carbon atoms,
The reaction is carried out in a solvent that does not react with these and, if necessary, in the presence of a catalyst.

The concentration of fatty acids having 6 to 24 carbon atoms, fatty acid anhydrides having 6 to 24 carbon atoms, and fatty acid chlorides having 6 to 24 carbon atoms is preferably 5% by weight or more, and the amount used is based on the weight of the gel. The same amount or more is sufficient. The reaction is -20°C
The process proceeds by heating at ~100°C for 1 hour or more.

The spherical particles having a particle size of 1 to 200 μm, preferably 1 to 15 μm, obtained as described above can be used as a carrier for ion exchange chromatography, and after further modification, can be used as a carrier for Affinity chromatography.

〔Example〕

Examples of the present invention are shown below. Hereinafter, % means weight %.

Example 1 (1) Synthesis of base gel which is a copolymer of glycidyl methacrylate and ethylene glycol dimethacrylate (
Polymerization reaction) 175 g of glycidyl methacrylate, 25 g of ethylene glycol dimethacrylate, 12 g of isoamyl alcohol
A mixture of 5 g, butyl acetate, 95 g, and azobisisobutyronitrile 1.0 g was suspended in 0.2% methylcellulose aqueous solution 22, and mixed for 1 minute at room temperature under high-speed stirring using a homomixer. , and then transferred to a common propeller stirring device and reacted at 80°C for 7 hours to obtain gel particles insoluble in water and organic solvents. This gel was collected by 5 filtration, 21 parts water, 2 parts methanol! After washing, it was air-dried. The glycidyl group fit (measured by the hydrochloric acid-calcium chloride method) of this gel was 5 meq/g.

(2) 100 g of gel particles obtained in the hydrolysis reaction (1) were mixed with 100 g of 0.5N sulfuric acid.
0d and reacted at 80°C for 5 hours. This was collected through 5 filtration, washed well, and then dried. The amount of glycidyl groups in this gel (measured by the hydrochloric acid-calcium chloride method) was Omeq/g.

(3) 50 g of gel obtained in reaction (2) with stearic acid chloride, 500 g of dehydrated distilled pyridine
g and 25 g of stearic acid chloride were mixed, kept at 10°C for 1 hour while cooling with ice water, and then heated to 60°C for 5 hours.
Allowed time to react. The generated gel was collected by filtration, washed well, and then dried. Yield was 61g.

Comparative Example 1 Example 1 except that 160 g of glycidyl methacrylate and 40 g of ethylene glycol dimethacrylate were used.
Polymerization and hydrolysis reactions were carried out in the same manner as above. Glycidyl group amount after polymerization reaction (measured by hydrochloric acid-calcium chloride method)
is 4.5 meq/g.

The amount of glycidyl groups after the hydrolysis reaction (measured by the hydrochloric acid-calcium chloride method) was On+eq/g.

Application example 1 The gel obtained in Example 1 was sorted to give 10 to 15
The particles had a particle size of μm and were packed into a stainless steel column with a diameter of 6.0 mm and a diameter of 150 mm. Figure 1 shows a chromatogram in which benzene, naphthalene, and anthracene were separated using this column. In FIG. 1, the symbols 1.2 and 3 indicate the elution volume peaks of benzene, naphthalene, and anthracene, respectively. The mobile phase was methanol:water (9
5:5), flow rate is 1.0j11/mm, detection is UV25
I went at 4r+m. Also, 100% water, 10% methanol
Even when flowing 0%, there were no voids in the column.

Application Example 2 The gel obtained in Comparative Example 1 was sorted and packed in a column in the same manner as in Application Example 1, and benzene, naphthalene, and anthracene were separated under the same conditions as in Application Example 1. The chromatogram is shown in FIG. 2, but benzene, naphthalene, and anthracene are not separated and cannot be used as a reverse phase packing material.

〔Effect of the invention〕

According to the method of the present invention, it is possible to provide a column packing material for reversed-phase partition chromatography that has excellent acid resistance and alkali resistance and has small differences in swelling and shrinkage between various solvents.

[Brief explanation of the drawing]

Figure 1 is a chromatogram obtained using gel particles produced according to Example 1 of the present invention as a filler, and Figure 2 is a chromatogram obtained using gel particles produced according to Comparative Example 1 as a filler. This is a chromatogram. Explanation of symbols 1...Benzene elution peak 2...Naphthalene elution peak 3...Anthracene elution peak distortion 1, ry) 0 2 4 Otsu g / θ Welding each 1 (Masu 1)

Claims (1)

[Scope of Claims] 1. Glycidyl groups of gel particles obtained by aqueous suspension copolymerization of glycidyl monovinyl ester or glycidyl monovinyl ether and polyvinyl ester of polyhydric alcohol or polyvinyl ether of polyhydric alcohol, It is characterized by hydrolyzing in the presence of an unreactive acid, and further reacting with a fatty acid having 6 to 24 carbon atoms, a fatty acid anhydride having 6 to 24 carbon atoms, or a fatty acid chloride having 6 to 24 carbon atoms. A method for producing a column packing material for liquid chromatography. 2. Liquid chromatography according to claim 1, wherein 50 to 97 mol% of glycidyl monovinyl ester or glycidyl monovinyl ether and 50 to 3 mol% of polyvinyl ester of polyhydric alcohol or polyvinyl ether of polyhydric alcohol are aqueous suspension copolymerized. Manufacturing method of column packing material for 3. The method for producing a column packing material for liquid chromatography according to claim 1 or 2, wherein the glycidyl monovinyl ester or glycidyl monovinyl ether is glycidyl methacrylate.