JPS6233549B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a hydrophilic carrier suitable as a packing material for chromatography and a method for producing the same.
In particular, in high-performance liquid chromatography, proteins and
Separate important water-soluble substances such as enzymes and nucleic acids,
The present invention relates to a hydrophilic carrier suitable for measuring the molecular weight distribution of natural and synthetic water-soluble polymers and a method for producing the same. Conventionally, hydrophilic carriers include
Cross-linked dextran, agarose, polyacrylamide, etc. are known. These gels have low adsorption to water-soluble substances and have played a major role in the field of biochemistry as carriers for gel filtration chromatography, affinity chromatography, electrophoresis, etc., and as means for analysis and purification. However, due to their low mechanical strength, these gels can only be used at low flow rates and are time consuming. Also, PH, ionic strength,
It is also known that changes in the eluent composition change the degree of swelling of the carrier, significantly reducing column efficiency. With the recent development of advanced liquid chromatography, there has been a long-awaited separation support that has mechanical strength that can be used at high flow rates and has high column stability against changes in the eluent used. Therefore, methacrylate gels, polyvinyl alcohol gels, and porous glasses have been developed and used, but they are inferior in the most important aspects of adsorption and separation ability, and have not been successful with neutral sugars and some water-soluble synthetic polymers. I'm getting it, but it's still not enough. In particular, the application of the above-mentioned gels to biochemical-related substances has been insufficient. Recently, JP-A-46-7296 proposed a support in which organosilane was chemically bonded in a multilayer layer by polymerizing organosilane, but the hydrophilic support shown here was Si-O-C. However, this bond is susceptible to hydrolysis in water, and the silane agent elutes during use, resulting in changes in the thickness of the polymerized layer, resulting in a decrease in chromatography reproducibility and column efficiency. This is undesirable for long-term use due to the occurrence of JP-A-48-22099 proposed a carrier in which organosilanes having organic groups with large atomic weights were chemically bonded in a monomolecular layer. Due to steric hindrance, the amount of silane bonding per unit area cannot be increased, and the surface of the carrier cannot be sufficiently covered. This is presumed to be the reason why the protein recovery rate is still insufficient. Furthermore, the performance of glycerolpropylsilane -bonded porous glass has been reported in J. Chromatog. It is presumed that the recovery rate of proteins, etc. is still insufficient because there is no. However, from these reports, etc., it can be seen that high speed is possible, and the adsorption properties are considerably improved from the recovery rate and elution amount compared to conventional hydrophilic carriers for high performance liquid chromatography. The present inventors conducted studies to further improve adsorption properties and develop a hard gel capable of increasing speed.
First, organosilane having a terminal epoxy group with a small molecular weight is chemically bonded to a porous carrier having silanol groups on the pore surface, thereby covering the pore surface densely with organosilane. By adding a hydrophilic substance to the epoxy group to form a hydrophilic organic layer, it was possible to further reduce adsorption compared to the above-mentioned hard carrier. However, in general, reactions with functional groups bonded to carriers do not increase the reaction rate compared to reactions in the gas phase or liquid phase. If a simple polyol is selected as the hydrophilic substance to be added, the reaction rate will be low and the hydrophilic organic layer formed will not be sufficient to reduce adsorption. Therefore, the hydrophilic substance to be added is preferably a hydrophilic organic substance having an epoxy group, and in order to form a sufficient hydrophilic organic substance layer to reduce adsorption, it is necessary to add a certain amount of epoxy or more in the molecule of the hydrophilic substance. It is necessary that the group be contained. The pore surface of the support obtained in this way is not only covered with individual organosilanes with large organic groups, but also organic substances bonded to the organosilanes bond to each other to form a polymer layer. Therefore, we have completed the present invention by discovering a hard hydrophilic carrier that almost satisfies the above-mentioned uses of the soft gel in terms of adsorption properties, is suitable for high-speed operation, has high resolution, and provides stable column performance. That is, the present invention provides a hydrophilic carrier in which the porous surface is covered with a copolymer of silanol groups and organosilane on the pore surface with Si-O-Si bonds and an epoxy group-containing organic substance, and a method for producing the same. It is. More specifically, a hydrophilic carrier characterized in that the silanol group Si on the pore surface of the porous carrier and the following general formula ()Si are bonded by Si-O-Si, general formula () {R ₁ ; Bonds of C-C, C-O, C-H and O-H or bonds of C-C, C-O, C-H, O-H and C-X (X; halogen atom) ( The halogen atom content is 20% by weight or less), and the Si
is an organic group consisting of a Si-C bond and having an atomic ratio of C and O (C/O) of 2.5 or less, R ₂ ; the same organic group as R ₁ , methyl group, ethyl group, methoxy group, ethoxy group, or hydroxyl group , R ₃ ; methyl group, ethyl group, methoxy group, ethoxy group, or hydroxyl group, m: an integer of 1 or more, n: an integer of 0 or 1 or more} and the silanol group on the surface of the pores of the porous carrier and the general formula () {R ₄ ; an organic group having an epoxy group and the total atomic weight of its constituent atoms being less than 150, R ₅ ; the same organic group as R ₄ , a methyl group, an ethyl group, a methoxy group, an ethoxy group, or a halogen atom; R ₆ ; Organosilane represented by methyl group, ethyl group, methoxy group, ethoxy group or halogen atom, R ₇ ; methoxy group, ethoxy group or halogen atom} in the presence or absence of a solvent inert to the carrier and the organosilane. , react at 20°C to 300°C to form Si-O-Si bonds, and then form C-C, C-O, C-H and O-H bonds or C-C, C-O, C-H, O
It is composed of -H and C-X (X; halogen atom) bonds (halogen atom content is 20% by weight or less), has an atomic ratio of C to O (C/O) of 2.5 or less, has a molecular weight of 1000 or less, and has an epoxy equivalent. (Molecular weight/number of epoxy groups) An epoxy group-containing organic substance of 300 or less is copolymerized with an organosilane-treated support in an inert solvent at -20°C to 200°C in the presence or absence of a catalyst. The present invention provides a method for producing a hydrophilic carrier characterized by the following. The present invention will be explained in detail below. The porous carrier used in the hydrophilic carrier of the present invention is made of silica gel, porous glass, diatomaceous earth, etc., and the silanol groups and organosilane on the pore surface are bonded with Si-O-Si, and the silanol group and organosilane on the pore surface are bonded with Si-O-Si. It is a hydrophilic carrier that has a silyl group or polysilyl group due to copolymerization, and the carrier surface is covered with a hydrophilic organic stationary phase resulting from alcoholic hydroxyl groups or ether bonds. In the porous carrier used in the present invention, the pore surfaces are densely covered with organosilane, and organic substances are bonded to the organosilane to form a polymer layer.
It is optimal to have at least 1.0 pieces/nm ² , preferably at least 1.0 pieces/nm ² . Further, the carrier may have any shape, but spherical carriers are particularly preferred for use in chromatography. Furthermore, the particle size of the carrier is 1
A carrier having a pore size of ~500 μm, preferably 2-200 μm, and an average pore size of 20 Å or more can be used. The organosilane having an epoxy group in the present invention is desirably one that can easily bond with the silanol groups on the surface of the pores of the porous carrier and can cover the surface of the porous carrier as densely as possible.
It is preferable that the molecule has 1 to 3 lower alkoxy groups or halogen atoms, 1 to 2 organic groups having an epoxy group bonded to the end, and the others are methyl or ethyl groups. For example, in the general formula () {R ₄ ; an organic group having an epoxy group and the total atomic weight of its constituent atoms being less than 150, R ₅ ; the same organic group as R ₄ , a methyl group, an ethyl group, a methoxy group, an ethoxy group, or a halogen atom; R ₆ ; Methyl group, ethyl group, methoxy group, ethoxy group or halogen atom, R ₇ ; methoxy group, ethoxy group or halogen atom}, especially γ-glycidyloxypropyltrimethoxysilane, γ-glycidyloxypropylmonomethyldichlor Examples include silane, γ-glycidyloxypropyldimethylmonochlorosilane, 3,4-epoxybutanemonomethyldichlorosilane, 3,4-epoxybutanetriethoxysilane, diγ-glycidyloxypropyldichlorosilane, and the like. In the present invention, when the organosilane is chemically bonded to the silanol groups of the porous carrier, the presence of a solvent is not an essential requirement, but from the viewpoint of stability and operability for uniformity of surface treatment, it is usually done in the presence of a solvent. It is advantageous to carry out the reaction. The solvent may be any solvent as long as it dissolves the organosilane and is inert to the porous carrier and the organosilane, such as ethers, ketones, esters, hydrocarbons, halogenated hydrocarbons, nitriles, and N-substituted amides. class, sulfoxide class, 3rd class
Particularly preferred solvents include ethers such as M-butyl ether, ethylene glycol dimethyl ether, diethylene glycol dimethyl ether, tetrahydrofuran and dioxane, hydrocarbons such as toluene, xylene and M-hexane, chlorobenzene, Examples include halogenated hydrocarbons such as dibromobutane, N-substituted amides such as dimethylformamide, diethylacetamide, and N-methylpyrrolidone, and sulfoxides such as dimethylsulfoxide. The reaction between the silanol groups on the surface of the porous carrier of the present invention and the organosilane is a known method, and the reaction rate is slow at low temperatures and the amount of organosilane bonded is not sufficient. On the other hand, at high temperatures, the stability of the epoxy group in organosilane decreases, and the subsequent copolymerization with the epoxy group-containing organic substance will not proceed. It is better. Furthermore, the amount of organosilane used in the porous carrier depends on the amount of silanol groups present in the carrier, but since the organosilane used in the present invention binds only 3.3 μmole/m ² at the maximum (surface area of the carrier) × (3.3 μmol/m ² ) or more of organosilane is sufficient. In the silane-treated carrier, as long as the porous carrier having a silanol group and the organosilane form a Si-O-Si bond, there is no problem even if the epoxy group is modified by ring opening. However, when the epoxy group is ring-opening modified, the compound to be bonded needs to have sufficient hydrophilicity so as not to cause hydrophobic interactions with various water-soluble substances, and the epoxy group-containing organic substance must have sufficient hydrophilicity to avoid reaction. In addition, the ring-opening reaction must be carried out in the amount necessary to sufficiently cover the pore surface with the hydrophilic polymer through the subsequent copolymerization reaction with the epoxy group-containing organic substance. It is necessary to leave the epoxy group. Therefore, the ring-opening modification rate of the epoxy group due to hydrolysis is at most 70% or less, preferably 60% or less, or the molecular weight is 200% or less.
Aliphatic polyols such as ethylene glycol, glycerin, erythritol, pentaerythritol, sorbitol or the general formula HO( _-CH2
Ring-opening accompanied by addition of polyethylene glycol represented by _-CH2 -O-) _oH (n=1 to 5) is preferred. Especially when performing ring opening by hydrolysis, H ₂ O
in the presence of sulfuric acid, hydrochloric acid, perchloric acid,
The silanized carrier may be impregnated with a protic acid such as phosphoric acid. When performing ring opening accompanied by addition of aliphatic polyol or polyethylene glycol, heat to 60°C or higher in the presence of the aliphatic polyol or polyethylene glycol and the presence or absence of a solvent, or ring-opening modification with a catalyst. can do. The solvent used in this reaction may be inert to the organosilane, the aliphatic polyol and polyethylene glycol to be added, and may include aliphatic or alicyclic ethers, ketones, esters, hydrocarbons, and halogens. Examples include hydrogenated hydrocarbons, nitriles, N-substituted amides, and sulfoxides. Particularly preferred solvents include ethers such as ethylene glycol, dimethyl ether, diethylene glycol dimethyl ether, and dioxane;
Halogenated hydrocarbons such as 1,2-dichloroethane, 1,2-dichloropropane, 1,1,1-trichloroethane, N-substituted amides such as N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide Examples include sulfoxides such as. Catalysts used in this reaction include protic acids such as sulfuric acid, hydrochloric acid, perchloric acid, and phosphoric acid; Lewis acids such as boron trifluoride, boron trifluoride etherate, and zinc boron tetrafluoride; Examples include metal halides such as stannic, titanium tetrachloride, and aluminum chloride, and Lewis acids such as boron trifluoride etherate are particularly preferred. Next, the method for copolymerizing the silane-treated carrier and the epoxy group-containing organic substance involves stirring the silane-treated carrier and the epoxy group-containing organic substance at a predetermined temperature and in the presence or absence of an inert solvent and a catalyst for a predetermined time. This is to react while doing so. At this time, in order to remove impurities bonded to silicon atoms, it is preferable to impregnate in an acidic aqueous solution at a predetermined temperature for a predetermined time and then wash with water, if necessary. Thereafter, the product is separated, washed several times with an inert solvent, and then dried under reduced pressure to obtain a hydrophilic carrier. This operation does not particularly affect the elution volume or recovery rate of the sample to be measured, but if a carrier without this operation is used for chromatography, it will affect detection by a differential refractometer.
Drift phenomenon often occurs at the beginning of use. In fact, when the carrier that had not been subjected to this operation was washed with distilled water, organic carbon was detected, but no organic carbon was detected on the carrier that had been subjected to this operation. The epoxy-containing organic substance used in the present invention must have sufficient hydrophilicity so as not to cause hydrophobicity and interaction with water-soluble substances. In addition, it is necessary to contain a certain amount or more of epoxy groups so that the pore surfaces are covered with a polymer layer through a copolymerization reaction. Furthermore, as the molecular weight increases, the internal volume of the carrier decreases, which is undesirable. Therefore, the epoxy group-containing organic substance in the present invention is composed of C-C, C-O, C-H, and O-H bonds, and the ratio of the number of carbon atoms to oxygen atoms (C/O) is 2.5 or less. , a compound with a molecular weight of 1000 or less and an epoxy group equivalent (molecular weight/number of epoxy groups) of 300 or less, preferably 200 or less (hereinafter referred to as an epoxy group-containing organic substance), or C-C, C-
O, C-H, C-X (X: halogen) and O-
Composed of H bonds, the ratio of the number of carbon atoms to oxygen atoms (C/O) is 2.5 or less, the molecular weight is 1000 or less, the epoxy equivalent (molecular weight / number of epoxy groups) is 300 or less,
It is preferable to use a compound (hereinafter referred to as an epoxy group-containing organic substance) having a halogen atom content of 200 or less and a halogen atom content of 20% by weight or less, preferably 15% by weight or less. Epoxy group-containing organic substances include ethylene oxide, glycidol, 1,2,3,4-diepoxybutane, diglycidyl ether, ethylene glycol (mono- to di)glycidyl ether, glycerin polyglycidyl ether, pentaerythritol polyglycidyl ether, sorbitol. Polyglycidyl ether, diglycerol polyglycidyl ether, polyethylene glycol (mono to di)glycidyl ether, (polymerization degree of polyethylene glycol 9 or less) polypropylene glycol (mono to di)glycidyl ether (polymerization degree of polypropylene glycol 3 or less), 1. 4-Butanediol (mono to di)
Examples include glycidyl ether, 1,3-propanediol (mono- to di)glycidyl ether and the like. The glycidyl ether compounds listed here are manufactured in a conventional manner, for example by reaction of the corresponding aliphatic polyol with epihalohydrin, but in the normal manufacturing process, by hydrolysis of the oligomer or epoxy group of the glycidyl ether compound. Compounds that have become glycol groups and compounds containing residual halohydrin groups due to insufficient opening reaction of halohydrin groups are produced as by-products, but these compounds are also preferred epoxy group-containing organic substances in the present invention, and mixtures There is no problem in using it as. Epoxy group-containing organic substances include 3-glycidyloxy, propylene chlorohydrin, ethylene glycol glycidyl chlorohydroxypropyl ether, glycerol diglycidyl chlorohydroxypropyl ether, diethylene glycol glycidyl chlorohydroxypropyl ether,
Examples include sorbitol triglycidyl bromohydroxypropyl ether, diglycerol diglycidyl chlorohydroxypropyl ether and the like. Catalysts used in the copolymerization of epoxy group-containing organic substances with organosilane bonded to the carrier of the present invention include protonic acids such as sulfuric acid, hydrochloric acid, perchloric acid, and phosphoric acid, boron trifluoride, boron trifluoride, Examples include Lewis acids such as etherate, zinc boron tetrafluoride, and metal halides such as stannous or stannic chloride, titanium tetrachloride, aluminum chloride, etc., but Lewis acids such as boron trifluoride etherate are particularly preferred. . Further, in this reaction, although the presence of a solvent is not an essential requirement, it is usually advantageous to carry out the reaction in the presence of a solvent from the viewpoint of uniformity of surface treatment and operability. The solvent may be inert to the organosilane or the organic substance to be added, and may include aliphatic or alicyclic ethers, ketones, esters, hydrocarbons, halogenated hydrocarbons, nitriles, sulfoxides, etc. I can give you. Particularly preferred solvents include ethers such as ethyl ether, diethylene glycol dimethyl ether, ethylene glycol dibutyl ether, tetrahydrofuran, and dioxane, 1,2-dichloroethane,
Examples include halogenated hydrocarbons such as 1,4-dibromobutane, 1,2-dichloropropane, and 1,1,2-trichloroethane, and sulfoxides such as dimethyl sulfoxide. In this reaction, the reaction rate is sometimes fast even at low temperatures, but it is generally slow and impractical; on the other hand, at high temperatures, various side reactions occur and are unfavorable for adsorption, so ℃
It can be carried out over a temperature range of The optimum reaction temperature is selected depending on the addition compound, the type of epoxy group-containing organic substance for copolymerization, the type of solvent, the type of catalyst, etc., but it is usually carried out in the range of 0°C to 150°C. The amount of the epoxy group-containing organic substance used for copolymerization is required to be at least equivalent to the epoxy group present in the carrier. Use more than twice as much. However, when carrying out in a solvent, it is not necessary to increase the concentration above the concentration at which gelation occurs with the epoxy group-containing organic substance alone. As explained above, when the same epoxy group-containing organic group as R ₄ is used as R ₅ in the general formula () as an organosilane, the epoxy group moiety of both R ₄ and R ₅ , that is, R ₁ in the general formula () Since copolymerization occurs at the and R ₂ parts, m and n in the general formula () are both integers of 1 or more. However, if another functional group is used as R ₅ , m in the general formula () becomes an integer of 1 or more. is copolymerized with an integer of , but the R ₅ portion, that is, the integer of general formula ()
Since copolymerization does not occur at the R ₂ portion, n is 0. Furthermore, the halogen atoms present in organosilane change to HCl when forming Si-O-Si bonds through reaction with silanol. Furthermore, the unreacted halogen atoms also react with water during subsequent contact with an aqueous solution and change into HCl, all of which replace hydroxyl groups. The carrier of the present invention has extremely superior performance as a carrier for high-speed gel filtration compared to known conventional carriers. Its first characteristic is low adsorption. This is because the pore surface of the carrier is covered with a hydrophilic polymer layer, so the hydrophobic interaction between the substance to be measured and the carrier surface is extremely small. Secondly, silica gel and porous glass are inorganic carriers that can be easily prepared using known methods as base materials for carriers.These carriers have a much narrower pore size distribution than organic polymer carriers, A hard gel with no change in degree of swelling due to differences in temperature can be obtained.
Therefore, the separation ability is very high, the speed can be easily increased, and the column stability is good. Further, although adjustment of the pore size depends on the carrier used as the base material, carriers with various pore sizes can be prepared using known methods. Therefore, the hydrophilic carrier obtained in the present invention has extremely excellent performance, and can be used to transport proteins, enzymes, polypeptides, amino acids, nucleic acids, nucleotides, sugars, vitamins, hormones, water-soluble synthetic polymers, etc. It is extremely effective when separating using a solvent using the molecular sieve effect. It can also be used for chromatographic measurements in other polar solvent systems, and various functional groups can be introduced using the hydroxyl groups on the surface. Can be used as a base material. Hereinafter, the present invention will be explained in more detail with reference to Examples. Example 1 The physical properties of the silica gel carrier used as the base material are: shape: spherical particle diameter: 10-12 μm; specific surface area: 250 m ² /g; average pore diameter: 250 Å. Place this silica gel in a flask equipped with a reflux condenser.
After drying at 60℃ and 10mmHg for 6 hours, take 24 parts (all parts by weight) and use γ- as _R4 in the general formula ().
Glycidyloxypropyl group

[Formula] 20 parts of γ-glycidyloxypropyltrimethoxysilane having methoxy groups as R ₅ , R ₆ , and R ₇ is impregnated into a solution diluted with 200 parts of toluene. The reaction is carried out under reflux for 8 hours while distilling off the methanol produced. Separate the silanized silica gel and add 3 parts with 100 parts of toluene.
Wash twice and dry under reduced pressure. Next, the total amount of the obtained silane-treated silica gel and glycerin polyglycidyl ether as an epoxy group-containing organic substance A solution prepared by diluting 10 parts with 200 parts of 1,4-dioxane is placed in a reactor equipped with a stirrer, and while stirring at room temperature, 1 part of boron trifluoride etherate is injected, and then the reaction temperature is raised to 80°C. and react for 2 hours. Next, wash three times with 100 parts of dioxane. Furthermore, in order to remove the alkoxy group bonded to the silicon atom, it was impregnated with 200 parts of aqueous hydrochloric acid solution of PH3,
After reacting at ℃ for 12 hours, the carrier was washed with 100 parts of water three times and dried under reduced pressure.
Wet-packed into a stainless steel column with a length of 600 mm.
The performance as a packing material for gel permeation chromatography using an aqueous solvent was investigated under the following conditions. The general formula () on the carrier obtained in this way is
R ₁ is Here, R ₂ and R ₃ are hydroxyl groups. Measurement conditions Equipment HLC-802UR (trade name of Toyo Soda Kogyo Co., Ltd.) Detector Differential refractometer and ultraviolet absorption detector (280 nm) Eluent Water or phosphate buffer (1/15 mol, phosphate buffer, PH6. 81/10 mol Kc) Flow rate 1 ml/min Sample injection amount 0.1 solution 100μ (protein) 0.2-0.5% solution 100μ (others) First, using water as the eluent, dextran (MW: 500000) (manufactured by Pharmacia, Dextran T -500) and ethylene glycol to determine the gel capacitance. (Gel capacity)=(Amount of ethylene glycol elution)-(Amount of dextran T-500 elution)/(Amount of dextran T-500 elution) At this time, the gel capacity was 1.40.
Next, the eluent was changed to phosphate buffer, and the elution amount (elution position) and recovery rate of various proteins were measured.
The results are shown in the table. To measure the recovery rate, calculate the peak area when passing through a stainless steel column with an inner diameter of 0.6 mm and a length of 7.4 m.
Total peak area (main peak + impurity peak) when passing through a packed column, assuming 100%
Expressed as a percentage. In this case, an ultraviolet absorption detector was used as the detector, and the measurements were performed three times and the average was determined. The results of compositional analysis in the dry state of the carrier after silane treatment and the carrier after copolymerization reaction in this example are shown below.

[Table] Furthermore, the infrared spectrum of the carrier obtained in this example is obscured by the absorption of the raw material silica gel itself, except for the absorption of paraffins at 2940 cm ^-1 and 2880 cm ^-1 . Therefore, this support was repeatedly heat-treated in an aqueous hydrogen fluoride solution to completely remove the SiO ₂ component, and then water and low-boiling compounds were removed from the aqueous solution using a rotary evaporator, and the residue was exposed to infrared rays. When the spectrum is measured, there is an aliphatic ether absorption at 1120 cm ^-1 , a hydroxyl group at 1050 cm ^-1 and 3450 cm ^-1 , and a paraffin absorption near 2900 cm ^-1 . Comparative Example 1 Using the same silica gel as in Example 1, it was silanized with the same organosilane under the same conditions, washed and dried, and the obtained silanized silica gel was impregnated with 200 parts of an aqueous hydrochloric acid solution of PH3, and heated at 40°C. at 12
A time reaction is performed to remove the alkoxy group bonded to the silicon atom and to hydrolyze and ring-open the epoxy group. After thorough washing with water and drying under reduced pressure, the obtained carrier was wet-packed into the same column as in Example 1, various proteins were measured under the same conditions, and the recovery rates were determined. The results are also shown in the table.

[Table] Examples 2 to 8 Using the same silica gel as in Example 1, various organosilanes, solvents, and epoxy group-containing organic substances,
Using a catalyst and a solvent, the carbon content of the carrier obtained after reacting under the same conditions as in Example 1 was measured. The results of determining the recovery rate of C) are shown in the table. In addition, the correspondence with the general formula of each compound in each example is shown in the table. In addition, glycerin polyglycidyl ether
The Cl content is determined by the fact that some of the glycidyl groups are -OCH ₂ CH
This is because it is the chlorohydrin moiety of (OH)CH ₂ Cl. Example 9 Before copolymerizing the organosilane-treated silica gel of Example 1 with an epoxy group-containing organic substance, some of the epoxy groups were immersed at 30°C for 2 hours in 200 parts of an aqueous solution adjusted to pH 5.5 by adding hydrochloric acid. The carbon content of a carrier obtained by treating ring-opening modified silica gel under the same conditions as in Example 1 and the carbon content of a carrier obtained by wet-packing this carrier into the same column and measuring proteins (human serum albumin, myoglobin, The results of determining the recovery rate of cytochrome C) are also shown in the table. The epoxy concentrations before ring-opening modification and after modification are as follows: Each dry gel is placed in a certain amount of dry dioxane in which hydrogen chloride is dissolved, and after the epoxy group of the gel is ring-opened to form chlorohydrin, the remaining hydrogen chloride is removed with water. Back titrate with sodium oxide. Determined using the method of Anal, Chem, 19 414 (1947). The result is 0.75 m before ring-opening modification. eq/g 〃 Back 0.55m. It was eq/g. Example 10 Glycerin was added to the organosilane-treated silica gel of Example 1 before copolymerizing it with an epoxy group-containing organic substance.
100 parts diethylene glycol dimethyl ether
The mixture is placed in a reactor equipped with a stirrer together with a solution diluted with 100 parts, and 1 part of boron trifluoride etherate is injected while stirring at room temperature of 25°C, and the mixture is allowed to react at the same reaction temperature for 1 hour. Next, 1,4-dioxane
The carbon content of the carrier obtained by washing the silica gel twice with 100 parts under the same conditions as in Example 1 is shown below. The results of determining the recovery rates of serum albumin, myoglobin, and cytochrome C) are also shown in the table. The epoxy concentration after ring-opening modification was determined in the same manner as in Example 9, and the result was 0.25m. It was eq/g. Comparative Example 2 Using the same silica gel as in Example 1, it was silanized with the same organosilane under the same conditions, washed and dried, and then the same operation as in Example 1 was performed without using the epoxy group-containing organic substance. The carbon content of the obtained carrier and the recovery rate of proteins (human serum albumin, myoglobin, cytochrome C) obtained by wet packing this carrier into the same column under the same conditions are also shown in the table.

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Claims (1)

[Scope of Claims] 1. A hydrophilic carrier characterized in that silanol groups Si on the pore surface of the porous carrier and the following general formula ()Si are bonded by Si-O-Si. General formula () {R ₁ ; Bonds of C-C, C-O, C-H and O-H or bonds of C-C, C-O, C-H, O-H and C-X (X; halogen atom) ( The halogen atom content is 20% by weight or less), and the Si
is an organic group consisting of a Si-C bond and having an atomic ratio of C and O (C/O) of 2.5 or less, R ₂ ; the same organic group as R ₁ , methyl group, ethyl group, methoxy group, ethoxy group, or hydroxyl group , R ₃ ; Methyl group, ethyl group, methoxy group, ethoxy group, or hydroxyl group, m: An integer of 1 or more, n: 0 or an integer of 1 or more} 2 Silanol group on the surface of the pores of the porous carrier and the general formula () {R ₄ ; an organic group having an epoxy group and the total atomic weight of its constituent atoms being less than 150, R ₅ ; the same organic group as R ₄ , a methyl group, an ethyl group, a methoxy group, an ethoxy group, or a halogen atom; R ₆ ; Organosilane represented by methyl group, ethyl group, methoxy group, ethoxy group or halogen atom, R ₇ ; methoxy group, ethoxy group or halogen atom} in the presence or absence of a solvent inert to the carrier and the organosilane. , react at 20°C to 300°C to form Si-O-Si bonds, and then form C-C, C-O, C-H and O-H bonds or C-C, C-O, C-H, O
It is composed of -H and C-X (X; halogen atom) bonds (halogen atom content is 20% by weight or less), has an atomic ratio of C to O (C/O) of 2.5 or less, has a molecular weight of 1000 or less, and has an epoxy equivalent. (Molecular weight/number of epoxy groups) An epoxy group-containing organic substance of 300 or less is copolymerized with an organosilane-treated support in an inert solvent at -20°C to 200°C in the presence or absence of a catalyst. The silanol group Si on the surface of the porous carrier and the following general formula ()Si
are combined by Si-O-Si {R ₁ , R ₂ , R ₃ , m, n; same as above} Production method of hydrophilic carrier.