JPS60106802A - Production of hydrophilic gel - Google Patents

Abstract

PURPOSE:To produce the titled gel which does not undergo nonspecific adsorption irrespective of pH, by introducing a polysaccharide having a number of hydroxyl groups in the molecule as a hydrophilic group into a styrenic hydrophobic polymer gel. CONSTITUTION:A PS forming a styrenic hydrophobic gel (styrene/divinylbenzene copolymer) is reacted with chloromethyl methyl ether in the presence of anhydrous stannic chloride to introduce chloromethyl groups into the benzene ring of styrene, and an alkali is reacted with the product in the presence of a phase transfer catalyst to convert the chloromethyl groups into hydroxylmethyl groups. A polysaccharide is reacted with the hydroxyl groups of the obtained hydroxymethylated polystyrene gel in the presence of anhydrous stannic chloride to bond the polysaccharide thereto through glycoside bonds. USE:Filler for high-performance liquid chromatography.

Description

[Detailed Description of the Invention] Technical details The present invention is based on the following method: 1. Adding water to a hydrophobic polystyrene gel suitable for comatograno using a high-speed liquid. 11-ni'C+'l;li
;;How to calculate Q: -1.

Conventionally, when analyzing water-soluble biopolymers, such as proteins, IL, and IIR, using liquid chromatography.

As the filling Fi11, a gel containing 7yp% polysaccharide is used. This gel is an excellent packing material with almost no non-specific adsorption, but since it is a soft gel, it has a low 1M resistance limit. Therefore, it cannot be used in high performance liquid chromatograph f-. For high performance liquid chromatography,
In order to prevent non-specific adsorption, a silica gel odorant with a silane surface treatment (chemically bonded silica gel) is used. JP-A No. 56-93043 discloses that the surface of a porous material such as silica gel is treated with a silane coupling agent having an epoxy group at the end such as γ-glycidoxypropyltrimethoxysilane. Then, the hydrophilic gel obtained by reacting sugar chains with this was used as a gel fill for chromatography such as protein%.
” The number of months to be used is indicated. However, since such a surface j4 agent is based on silica gel, it has an acidic
It can only be used under certain conditions, and it is impossible to use pH9 at pH 8 or higher.

Excellent stability in both acidic and alkaline conditions.
In addition, there is a polystyrene gel of styrene-divinylbenzene copolymer that can be used in high-performance liquid chromatography. However, this gel has strong hydrophobic interactions.

4. For direct protein intake, etc. ), non-specific adsorption occurs. Therefore, 1!J of ion replacement with an ionic derivative and an absorbent can only be used for hydrophobic chromatography.

Purpose of the Invention The present invention [1] Preparation of 11 for packing for analyzing water r=I・'t, H% molecules such as protein '1p and Kuwaken existing in living bodies by high performance liquid chromatography. The purpose is to provide a method. 41 of the present invention! The purpose of i is
The object of the present invention is to develop a method for producing a one-piece fillet for tomatogelfish, which has a high pH value and does not cause non-specific adsorption.

Nephew of the Invention The method of making water leaf gel according to the present invention is as follows: 1. Styrene-based hydrophobic polymer gel coated with high performance liquid chromatography (m) and 1++1. From Tojin C, which has many hydroxyl groups in the molecule as aqueous groups (1' = 72),
Hydrophilic game Jl/l Koryu lli! This method involves the process of inserting a hydroxymethyl group into the benzene ring of styrene that forms a styrene-based hydrophobic polymer gel, and reacting the hydroxymethyl group with a sugar to valve the glycosidic bond to form the paulownia 4. It has a finalizing process.

Does that lead to the above purpose? will be accomplished.

Polystyrene, which is widely used as a styrene-based hydrophobic six-molecular gel, is a styrene-divinylbenzene copolymer. When this is reacted with chloromethyl methyl ether in the presence of anhydrous stannic chloride.

As shown in the formula below, the chloromethyl group is exclusively present in the benzene ring of styrene. Furthermore, when an alkali is applied in the presence of a phase transfer catalyst, it is converted into a hydroxymethyl group.

CH,OH The hydroxyl group of this hydroxomethylated polystyrene gel is reacted with a saccharide in the presence of anhydrous ``4'', ``ni'', and ``ni'' to give glycoside 6'7 and 1,6. For example, to introduce .beta.-D-glucose, pentaacetyl-.beta.-1)-glucose is reacted to form glycoside 6'7, as shown in the following formula.

Ac Then, as shown in the following formula, C is converted into a carbon using an alkali to remove the acetyl group, which is the first → group.

Example The present invention will be explained below using a preferred embodiment G.

Hydroxymethylation of styrene-based hydrophobic gel: 50 g of polystyrene gel (styrene-divinylbenzene copolymer; spherical, average particle size 10 ttm) dried under reduced pressure overnight was placed in a 1-sized separable flask, and dried chloroform 3
This was stirred at room temperature for 2 hours. This was cooled to 0°C, and 25% of chloromethyl methyl ether was mixed with 25% of stannic chloride.
It was added together with m and allowed to react for 1 hour. Dissolve the reaction mixture. Separately, add the gel to a dioxane-water mixture (3:IV/V
), groove cleaning with dioxane-diluted hydrochloric acid mixture (3:IV/V). Pure water was gradually replaced with dioxane. Reduce the volume of chloromethylated styrene gel overnight at 6 ()°C.
lf dried. Its collection is 4 6, 7 g. The chlorine concentration was 5.6 times Ti'c (flask smoking method - nitric acid Akirabe method). 20 g of the obtained chloromethylated styrene gel was added to 150 mi of 0-dichlorobenzene.
% cloudy. An aqueous solution (55 mL) containing 28 g of potassium acetate and 9 g of potassium hydroxide was added thereto, followed by the addition of porridge and 6 g of tetrabutylammonium hydroxide.

This was stirred vigorously at 80° C. for 24 hours until the mixture was dissolved.

After the reaction, the gel was distributed from door to door.

Methanol, water, detrahydrofuran-water mixture (3:)
V/V), tetrahydrofuran, acetone.

Washed with dichloromethane and methanol. This 11 was dried under reduced pressure overnight at room temperature. Its yield is 18.2
11) ivy in g.

(Bl Introduction of 1 octopus into styrene-based hydrophobic gel: 10 g of hydroxymethyl styrene gel.
-'r dichloromethane 80' mJ2 (l C 30
Minute IH mouth ゛ early cloudy sasebuko. 'Penta-]'Cetyl-β-[) //'')Recose (Song Tunhua 1+, I
(manufactured by Ill.) and 12 g of anhydrous stannic chloride were dissolved in 150 ml of dichloromethane with OC, and this was added to the solution of styrene 6 gel.

41L' in OC? Song IV 'I made it to the core. gel from door to door;
Methanol, 4<, ? ``+'', 1.ii Acid, saturated water, methanol-water mixture (1:1) was added to 'l!i'l and washed.The obtained gel was washed with
V tatami・■・S14-ta. The yield was 10g.

Absorption of ester (1750I71+') was confirmed by infrared, early spectrum (KI3r disk method). 10g of gel was added to 200 g of methanol-water mixture (1:1).
Add to a solution of 5 g of potassium hydroxide dissolved in ηl,
It was treated at 0°C for 2 hours. The gel was washed separately with water and methanol. Next, the mixture was suspended in water-methanol (1:1) and dispersed in an ultrasonic cleaner for 5 minutes. Then, the part that did not precipitate was removed by decantation. Collect the precipitated gel.

-I made Kensuke Yasumuda. The yield was 9.0g.

Infrared spectroscopy revealed that the ester yield had disappeared. Absorption based on hydroxyl group is 330
0 t: m-- Make sure that the light is shining on the column A7 to (Q)
Add 20g of the hydrophilic Ihigel obtained in Section 11 to a water-methanol mixture (1:IV/V)! It was cloudy. this! 1■
The turbid liquid mode was set to 70 Lμd), and water was used as a feed liquid to fill a stainless steel column with a diameter of 7.9 M and a length of 500 mm. After 300 tni of King's solution was poured, it was removed from the filling machine 1 and connected to the Takahyaku Shu liquid chromatography apparatus 6 for analysis. As samples, maltose, maltobeki-11/-sugar bovine serum albumin (BSA),
1! Liethylene glycol (1νlW6 jl (1)
The printing line 11r (i 1ftl) of each sample was determined using an aqueous solution of 1111. The results are shown in the table below.

The pump was a 1'' manufactured by Honbunko (Tri) and 0 tarV (G). % syneresis is pHoto, IM) 1.
ml, 7+ij at lZf speed of 7 minutes. For run 4, a differential 1i'i, l 4 loaf meter made by Waters was used.

17-ri intersection example ゛1, Q hydrated gel 1 (instead, polystyrene gel was packed in Shio II 1 (Q JlN and 11 times, put it on the ζ column, and tried each of the following 1 and 4 times)・1's I'r l i11
J: ``I've decided on the song.I'm going to put that Jokichi style on -F.''

According to the method of the present invention, a hydrophobic styrene polymer gel S, which has a plurality of hydroxyl groups as an aqueous group, is converted into a hydrophilic gel. Therefore, the white matter that exists in the living body? 1 (High-speed analysis of water-monolytic polymers such as moe / l [enables analysis by body chromatography. In the case of KEL, it is possible to make a gel for aqueous gel permeation chromatography by introducing a 4-aqueous group.

Furthermore, the spring-lasting polymer gel that is rM in by this method has a wide %N pH regardless of the liquid property of the sample τ solution.
It can be used with other members of the group, and has the critical point of not causing different adsorption.

that's all Publisher: Bond Suikado Koei Co., Ltd.

Claims (1)

[Scope of Claims] 1(1) A process for introducing a hydroxymethyl group into the beno(yne) ring of styrene forming a styrene-based hydrophobic VL polymer gel; A method for producing a Vl:J, tJ<> gel, comprising: reacting saccharides and linking the nine rivers through glycosidic bonds.