JPS6096605A - Production of hydrophilic resin - Google Patents

Abstract

PURPOSE:To produce a hydrophilic resin suitable as a packing of gel permeation chromatography, by subjecting a glycidyl group-containing crosslinked copolymer to a ring opening reaction in a solvent which can swell the above copolymer. CONSTITUTION:As a solvent for a ring opening reaction, acetone, 1,4-dioxane, dichloromethane, toluene or the like is used. Namely, (A) a glycidyl group-containing crosslinked copolymer (a copolymer of a glycidyl ester of an unsaturated carboxylic acid with a polyvinyl compound) is added to (B) about 5-20ml, per g of component A, of the above solvent and swollen, and after adding (C) polyethylene glycol (MW>=about 200) or glycerin and (D) about 5-50mg, per g of component A, of a catalyst (e.g., sulfuric acid), the resulting mixture is reacted by ring opening at about 50-100 deg.C for about 3-10hr to render the crosslinked polymer hydrophilic.

Description

DETAILED DESCRIPTION OF THE INVENTION Specifically, the present invention relates to liquid chromatography, particularly for separating water-soluble substances having a molecular weight of less than /θ0θ, such as oligosaccharides, oligopeptides, and shield proteins.

Conventionally, the optical filler for Kölpermeation chromatography is crosslinked dextran, which is obtained by crosslinking dextran I with epichlorohydrin or the like.

Kake 1i is a compound of agarose or acrylamide and methylenebisacrylamide! 3 Polyacrylamide etc. are used.

However, in the fillers of Nos. 1 and 1, the cross-linking pattern 1 is reduced and the gel network + h structure is widened (7), so the particles are soft and have low mechanical strength.

Therefore, there is an active yIE point where the particles are deformed during chromatographic operation under pressure and a pressure drop occurs.

On the other hand, in order to improve this tArb point, glycidyl ester of unsaturated carboxylic acid is crosslinked with ethylene glycol dimethacrylate, etc., and the epoxy group of the copolymer is combined with water or lower alkylene glycol, polyalkylene glycol, polyol 11'i. Diluted with a denaturing agent/Gel Permini/Yon Chromatography J (For example, JP-A-53-/θ and 2-11 (t
) L Kashi, Nakara.

Although this product has improved strength Q: 1, the difference between the elution capacity of a 1 molecular weight substance and the elution capacity of a lower limit molecular weight substance is not a light minute. Therefore, when the fractionation range of a substance is constant, the retention capacity increases as the retention capacity increases. The ode becomes gentler,
Considering that the distance between adjacent peaks becomes longer and the separation is improved, the packing material is still not sufficient. In gel permeation chromatography, which synthesizes water-soluble substances with a molecular weight of 1000 or less in descending order of molecular weight, we are looking at a method for producing a water-absorbing resin with good separation properties and high mechanical strength. and arrived at the present invention.

The present invention consists in a method for producing a hydrophilic resin by subjecting a crosslinked copolymer having a glycidyl group to a ring-opening reaction with polyethylene glycol or glycerin.

The present invention will be explained in further detail below.

The cross-linked co-oxygen having a glycidyl group of the present invention can be obtained by combining a glycidyl ester of an unsaturated carboxylic acid with a polyvinyl compound as a cross-linking agent by a conventional method 1, for example, in JP-A-53-7
11 is produced by copolymerizing according to the methods described in No. 0 and No. 7.

Examples of glycidyl esters of unsaturated carboxylic acids include glycidyl acrylate, glycidyl methacrylate, glycidyl crotonate, diglycidyl itaconate, diglycidyl fumarate, and diglycidyl maleate. On the other hand, as a polyvinyl compound as a crosslinking agent, carbon n2-3 carbon glycol i k Ir
e'.

Esters of these polyalkylene glycols and acrylic acid or methacrylic acid, for example.

Acrylic acids of polyols such as ethylene glycol diacrylate, ethylene glycol dimethacrylate, propylene glycol diacrylate, propylene gelicol dimethacrylate, polyethylene glycol dimethacrylate, polyethylene glycol dimethacrylate, polypropylene glycol diacrylate, polyglopylene glycol dimethacrylate, etc. Also, methacrylic acid ester, divinylbenzene.

Divinyltoluene, divinylxylene, divinylethylbenzene, divinylnaphthalene l1, etc. p1゛.

Equal parts of aromatic compounds are used.

The polyvinyl compound is j (weight)% of the total polymerizable monomers.
As mentioned above, it is wisely used so that it is /j (weight)% or more.

As a diluent during polymerization, it is a solvent for polymerizable monomers)
1) Use something inert to the polymerization reaction.

Further, since the hemopolymerization is usually carried out by a suspension-polymerization method, the diluent must be insoluble or print-soluble in water, which is a polymerization dispersion medium.

Such diluents include aliphatic compounds such as dichloromethane, dichloroethane, trichloroethane, etc., dimethyl phthalate,
Aliphatic or aromatic esters such as diethyl phthalate are included.

A diluent is a combination of a polymerizable monomer and a dilution series.

Against 7, at least 30 vol.
is used so that the relationship is 41o~ and θ capacity i1;.

Examples of polymerization starting material include peroxides such as benzoyl peroxide, t-butyl hydroperoxide, lauroyl peroxide, dimetahydroperoxide, methyl ethyl ketone peroxide, or azobisin butyl peroxide. Lonitrile, Asohis α, α-dimethervanlonitrile, 2. ,．． 2'-Azobis-11,
Goodimethylvaleronitrile, azobis α-methylbutyronitrile.

Azobisinbutyramide Azo, 11 offshore start blue I
J is used. Its use l, 0 of the polymeric monomer
．． 7~/0 (r, Ii: 1li).

In terms of polymerization, jlj-formable monomer is polymerized and dispersed;
In order to prevent these substances from dissolving in water, substances such as sodium chloride, calcium chloride, etc. may be dissolved in water. Also, to stabilize the dispersion, use gelatin and polyvinyl alcohol.

Can be used in addition to a dispersion stabilizer such as carboxymethyl cellulose in all water.

The amount of the aqueous layer is approximately equal to or more than the amount of the organic layer and is not particularly limited, but it is used up to about /θ times the volume.

The polymerization reaction usually takes place at jθ~ and o°C at 3~! Separate them by an appropriate method such as applying time line processing or the like.

The polymer having a glycidyl group obtained as described above is made hydrophilic by an open-field reaction with polyethylene glycol or glycerin, and is reacted in the presence of a solvent that swells the copolymer. Yoshi1 molecular weight/θOθ
Easy production of water-based resins with good separation properties and high mechanical strength using gel permeation chromatography, which separates the following water-soluble substances 'i-1 in descending order of molecular weight. can do.

The polyethylene glycol used to ring-open the glycidyl group of a crosslinked copolymer having a glycidyl group usually has a molecular weight of 200 or more. Further, as the glycerin, any ordinary commercially available product may be used.

The solvent that swells the copolymer used in the ring-opening reaction usually has the property of dissolving polyethylene glycol and glycerin, and examples thereof include baths that dissolve linear glycidyl polymers. Specifically, acetone,
C3 to C9 lower aliphatic cutones such as acetonyl acetone, diethyl ether, C2 to C3 dioxane
Examples include low C4 aliphatic ethers or cyclic ethers, halogenated aliphatic hydrocarbons such as dichloromethane and dichloroethane, and aromatic hydrocarbons such as toluene, benzene and quinolene.

In the ring-opening reaction of the glycidyl group of a crosslinked copolymer with glycidyl groups, if the above-mentioned solvents are used, the ring-opening reaction may be caused by simply hydrolyzing with water, polyalkylene glycol or polyol, etc. The degree of swelling of the copolymer is preferably larger than that of the reacted one. For example, the degree of crosslinking when using a glycerin solution of /, \-dioxane is Jj%. The degree of swelling of the copolymer is based on the volume in glycerin. If it is 80.

23゛body concentration% concentration is 8/, jθ body concentration is 83.2j
With volumetric power /,! , 100% actual gravelness (semi-/, goodioxane) is 7 times larger than that, and swelling starts at 25% volume or more.

The amount of solvent used varies depending on the size of the solvent, but in terms of -1 dose, it is 5 to 20 mL per 72 copolymer, and the amount is 2 to 20 mL per total amount of polyethylene glycol or glycerin and solvent.
5 volume coefficient or more is preferred; Can the solvent be mixed with polyethylene glycol or glycerin? The copolymer may be reacted with the copolymer as a liquid, or the copolymer may be swollen in advance with a 6♀ medium and then reacted with polyethylene glycol or glycerin.

The ring-opening reaction of the glycidyl group is carried out in the presence of the above solvent.

The reaction is carried out using an acid catalyst frequently. For example, in sulfuric acid, copolymer/7 and j ryg
~,, using about 5' o my amount, jθ°C ~ 700°C,
The 3 to 70 o'clock reaction can selectively carry out the ring-opening reaction for hydrophilization without selectively cleaving the ester bond of the cross-linked co-111' compound.

The reaction mixture was cooled and then purified with pure water to obtain a filtrate.

The hydrophilized resin obtained by the method of the present invention has an ino (molecular weight (/
The water-bathable substance J+ of θ00 or less) was used as a filler for the purpose of gel permeation chromatography.
'4 has a large capacity h'5, and it has a high power and a high division layer 1 (sexuality).

As a matter of fact, there is nothing specific to the following examples.

Fugan Example/ (a) Frame 4 for curing glycidyl ester groups:, iy Production of co-merged particles What are ethylene glycol dimethacrylate 7jfl and glycidyl methacrylate L? ,/,,2
-dichloroethane 322 and 2.27-abbis-
Add a mixture of 2/3 ml of goo-dimethyl nitrile to demineralized water, and 2/00 ml of polyvinyl alcohol.
2, sodium chloride, and 4t2, and suspension polymerization was carried out at 1° C. for about an hour while stirring at high speed.

After cooling the reaction product, the generated co-11 combined particles were collected and washed with water. This copolymer was then mixed with toluene//:
zsrw and 373 ml of water at room temperature.
After stirring for an hour, filter. Furthermore, this copolymer/
! After repeating the process of adding it to l of methanol and stirring once! θ℃, and the time tag is 2.

The particles that have undergone the above operations are sieved and have a particle size of l~/2μnt.
Copolymer particles 1009 were obtained (degree of crosslinking: 21 degrees).

(b) The copolymer particles 207 obtained by the ring-opening reaction (a) in a polyethylene glycol solution were mixed with ICA in Sidiokidun/θθ at room temperature for about 20 hours. be defrauded. Next, 100 ml of polyethylene glycol (average molecular weight: Otsu 00) was added and mixed well.
Add.

? It can be heated with stirring for 11.7 hours at θ°C.

After cooling, the hydrophilic resin was soaked with pure water. j+ and
A hydrophilic compound that is filtered and bound in a polyethylene glycol solution is prepared.

The above hydrophilic resin was classified into particles with a particle size of ~70 g, an internal diameter of 7.3-Tn, and a length of θOmm in a stainless steel column with a light Jp L column. For known polyethylene glycol and ethylene glycol θ, ta% aqueous solutions, temperature -2J-1:,
Measured at a flow rate of O, ml/mm (7. When a calibration curve was created with the logarithm of the molecular weight on the vertical axis and the bath volume on the horizontal axis, the first
It became like the figure. In addition, the values for each elution city of 1 molecule @ 200 and O 0θ in polyethylene glycol of known molecular weight are taken as the average value of the M elution curve. From the calibration curve, the retention capacity between ethylene glycol (molecular weight: 6) and polyethylene glycol with a molecular weight of /x/θ3 was 7 ml.This packed column was used to hold 15 ml of polyethylene glycol solution with an average molecular weight of 20 (:). As shown in Figure 1, the effective retention capacity (capacity between the beginning and end of the chromatogram) is 3.9.
It was Go. First, peak detection was performed using refractive index.

Human example υ;j ring reaction example in glycerin bath solution/
- Using copolymer particles obtained in the same manner as in (a), 'A=
Example 7/u/--Producted in exactly the same manner as in (b) except that glycerin was used instead of polyethylene glycol,
A hydrophilic resin to which glycerin is chemically bonded (!-17
c. Similar to Example/-(c), this hydrophilic resin was classified 1-7, l~/θ/(/E
The inner diameter of the particle is 7j'1lll11, the length is θor,
+rn stainless steel column). Using this column, molecules -: 11 known polyethylene glycol and O-ethylene glycol were precipitated from an aqueous solution of θ, l to prepare a test sample, as shown in Figure 3. :Line yoshini~/X103 molecule: jj'fi''
jlL4 no Yasumo) Tani: I: "Ke,!, it was 9 ml.

Using this element column, polyethylene glycol with an average molecular weight of 2θ0 was divided into 7υ[shi].

However, human power i example/ is the same as (,・ζyo〈minute 1411sa force/ζ.

Example 3 Glycidyl methacrylate λg 02, ethylene glycol dimethacrylate 1-name 02, /, 2-dichlor on tank 3 recognition 7 and λ2.2. '-Azobis--, turn methyl chloride + 1 nitrile 12 was polymerized with a homogeneous mixed solution of 12 nitriles (4': i<- Example/--A frame 4i!!i J'- Producing polymer L-fi
i, copolymerized particles/θθ7 of 2 μm (degree of crosslinking, 20%).

Using the obtained copolymer particles, a hydrophilic resin to which glycerin was chemically bonded was prepared in exactly the same manner as in Example/-(b) except that glycerin was used instead of polyethylene glycol. It's Kotobuki.

A 0.6% aqueous solution of polyethylene glycol and ethylene glycol of known molecular weight was packed in a stainless steel column of ., 600 In1l+ and analyzed.7 A calibration curve was drawn and the result was as shown in Figure 1.

According to the calibration curve, the retention answer 11 for the molecular weight J1 of the molecular weight J1 of '8 amount is g. Om! It also had good disciplinarity.

Comparison Example/Example/-The co-merged Rt particles 209 produced in (a) are placed in θml of a 0.7N% aqueous solution containing gas dioxane and mixed well. In the evening, I took a bath in the water
Heat to °C for 3 hours with stirring (47 mm, length O θ0
(Filled.This casino all phases, min-1-, t+t
If known polyethylene glycol and 1 liter of ethylene glycol water, O, S ratio <, dissolved acid, 1 min 4 min.
-13 When I created the ship, it looked like this in Figure 3. Detection; 11th line" - Riots~/x/θ3 numerator 111"
The retention valley of the range +, 7 i, j, t, θ+nl was met. Using this column, polyethylene glycol with semi-uniform molecular I11 and θO can be obtained by dividing the amount of water-soluble substance with molecules -',,; IJ”rf4 Sanakatsume ζ on 11.

This is slightly smaller than the retention 'a-iii, 'Vi-'-, Ome and the X-mizu 1g-ijJ fat of the present invention.''11
However, the r;11 water-based resin of the present invention is more hydrophilic than the aqueous resin.
j:v) H-;1 ipa) how low 15, minute tono 11
People think that my sexuality has deteriorated.

[Brief explanation of drawings]

Figure 1 is an example (filled with hydrophilic fragrant resin for 1 minute - calibrated using a known polyethylene glycol solution and an aqueous solution of ethylene glycol). Figure 2 is a chromatogram obtained by analyzing polyethylene glycol with an average molecular weight of 200 as a sample with the packed column of Example/4. For a column filled with aqueous resin, a test was carried out using known polyethylene glycol and ethylene glycol water bath numbers.
(This is a NJ diagram.
Shi,. This is a diagram showing a test using polyethylene glycol of known molecular weight and a water bath solution of ethylene glycol for this column. FIG. 5 shows the hydrophilic resin obtained at the ratio 1i;, 9-1+'u/. FIG. 2 is a calibration curve prepared for a column obtained using polyethylene glycol of known molecular weight and a water bath solution of ethylene glycol. No. 3121 is a chromatogram obtained by using polyethylene glycol with an average molecular weight -○○ as a sample and dividing it in a packed column with a ratio jis/. In Figures 1 to 1, the vertical axis represents the molecular weight.
+11 ke bathing capacity (me). Applicant: Representative of Sannauka no Ebisu Kogyo Shishiki Company Person: Ben 1111 Practitioner, Hase-yo - wa power)/Name section 1 diagram shi? )1 to i22? Ji! Quantity 1' no η/) - Force 2 Nagimaride - t (tnl') - Me3 figure'iJ 岑子 ん whisper) Output ear amount (v p'n - difference! \ Ichikata Rin - Katade Perforation 1 (rnl) - Figure ＼ ＼ 15 /b

Claims (1)

[Claims] (1) Copolymerization is carried out in producing a hydrophilic resin by subjecting a crosslinked copolymer having a glycidyl group to a ring-opening reaction with polyethylene glycol or glycerin.