JPH0117412B2 - - Google Patents

Description

[Detailed description of the invention]

(a) Technical Field The present invention relates to a novel adsorbent, and particularly to an adsorbent comprising a crosslinked polymer containing a component having excellent interpolymer complex-forming performance as a constituent component. (b) Prior art It is well known that high molecular weight polymers are used as adsorbents, and in particular, ion exchange resins are used to separate and purify substances by adsorbing and desorbing specific ionic substances. It is also well known that inter-polymer complexes formed by hydrogen bonds undergo reversible inter-polymer complex formation and dissociation due to PH, and as a result, physical properties change significantly. At present, it has not yet been possible to separate substances using the method. The present inventors have recently discovered that this change in physical properties of interpolymer complexes can be utilized for substance separation, and
To obtain a carrier that easily adsorbs and desorbs the substance to be separated,
We found that it is essential to introduce an inter-polymer complex structure into a cross-linked polymer, and obtained an adsorbent made of a cross-linked polymer containing such an inter-polymer complex-forming component. It is something. Up until now, polymers in which interpolymer complex-forming components have been introduced into crosslinked polymers have so-called interpenetrating network structures in which polyoxyethylene is crosslinked with triisocyanate and then polymerized by adding acrylic acid and a crosslinking agent. IPN) [Odaka,
Polymer Journal, 14 , P985 (′82)]. (c) Problems to be solved by the invention However, in this IPN structure, both ends of the polyoxyethylene (POE) chain are fixed.
There are few dynamic changes in POE chain motility;
The inter-polymer complex formation-dissociation phenomenon that occurs here is not sufficient to be utilized as a difference in the physical and chemical properties of separation carriers (adsorbents). (d) Means for Solving the Problems As a result of intensive research into the introduction of inter-polymer complex-forming components into crosslinked polymers for the purpose of use as separation carriers (adsorbents), the present inventors found (A ) Polymerizing a compound that simultaneously has a polyoxyethylene (POE) chain and a polymerizable double bond in the molecule, (B) a carboxylic acid (derivative) having a polymerizable double bond, and (C) a crosslinking agent. It is possible to introduce an inter-polymer complex-forming component into a cross-linked polymer, and the resulting cross-linked polymer has a new chemical structure in which POE chains are grafted. The inventors have discovered that this is an excellent adsorbent, and have arrived at the present invention. That is, the present invention provides (1) general formula (), (In the formula, each R may be the same or different and represents H or an organic group, and l, m, and p are integers of 1 or more.) A crosslinked polymer containing an interpolymer complex-forming component represented by Adsorbents made of polymers, and (2) A. Compounds that simultaneously have a polyoxyethylene chain and a polymerizable double bond in the molecule, B. Carboxylic acids or derivatives thereof that have polymerizable double bonds, C.
General formula (), characterized by reacting with a crosslinking agent, (In the formula, each R may be the same or different and represents H or an organic group, and l, m, and p are integers of 1 or more.) A crosslinked polymer containing an interpolymer complex-forming component represented by This invention relates to a method for producing an adsorbent made of a polymer. (e) Effect of the invention The polymerization reaction in the present invention involves the use of A. a compound having a polyoxyethylene chain and a polymerizable double bond in the molecule at the same time, and B. a carboxylic acid (derivative) having a polymerizable double bond. While polymerizing,
Proceeds through matrix polymerization that forms an intermolecular complex with the POE chain, producing blocks derived from carboxylic acid (derivatives) containing double bonds that correspond to the POE chain, and these are crosslinked with a crosslinking agent. For example, a crosslinked polymer having the following structure can be obtained. (l, m, n, and p are integers of 1 or more.) In this way, in the crosslinked polymer obtained by the present invention, only one end of the POE chain is fixed, and the other end is free. The POE chain of this graft type and the carboxyl group based on the double bond-containing carboxylic acid (derivative) of (B) constitute an inter-polymer complex-forming component,
Compared to the graft type POE chain mentioned above, which has both ends fixed, the POE chain has remarkable mobility due to the formation and dissociation of an inter-polymer complex due to pH change, and this is effective for adsorption and separation of substances. It becomes. (f) Specific disclosure of constituent elements of the invention Regarding the compound used in the present invention to introduce an inter-polymer complex-forming structure into a crosslinked polymer, polyoxyethylene chains and polymerizable double bonds are It is a compound (B) that forms an inter-polymer complex after polymerization with the compound (A) contained within and its polyoxyethylene unit, and theoretically any such compound can be used. However, (A) is methoxypolyoxyethylene (meth)acrylate Methoxypolyoxyethylene vinyl ether CH ₃ O (-CH ₂ CH ₂ O) - _o CH=CH ₂ n=1-45 Polyoxyethylene (meth)acrylate (NOF; Blenmar PE90, PE200, PE350), etc. (B) includes carboxylic acids such as acrylic acid and methacrylic acid, as well as carboxylic acids such as polyitaconic acid monomethyl ester, maleic anhydride, and itaconic acid monomethyl ester. Examples include acid derivatives. In particular, methoxypolyoxyethylene methacrylate (acrylate) having an oxyethylene unit (p) of 100 or less, and methacrylic acid or acrylic acid that forms an interpolymer complex with the polyoxyethylene unit after polymerization are preferably used. The ratio of addition of (A) and (B) is determined by the addition ratio of oxyethylene units and carboxylic acids (derivatives) having polymerizable double bonds that have the ability to form inter-polymer complexes after polymerization with the oxyethylene units such as methacrylic acid or acrylic acid. ) is 0.1 to 10, preferably 1. Depending on the types of (A) and (B) above, the interpolymer complex-forming component of the following general formula contained in the adsorbent of the present invention R and R' in the formula represent H, an alkyl group such as methyl or ethyl, or an organic group such as an aryl group;

[expression] or the other

It may also be combined with [Formula] to represent an acid anhydride residue. As for the crosslinking agent (C), the following compounds having two or more radically polymerizable double bonds in the molecule can be used. Namely, they are ethylene glycol dimethacrylate, polyethylene glycol dimethacrylate, divinylbenzene, triallyl isocyanate, etc., and the amount added is 0.1 to 50%, preferably 0.5 to 30%, based on the monomer ratio. Examples of radical polymerization initiators include azoisobutyronitrile, t-butyl hydroperoxide, cumene hydroperoxide, benzoyl peroxide, acetyl peroxide, di-t-butyl peroxide, azodicyclohexylcarbonitrile, dimethyl azodiisobutyrate, and succinic acid. Peroxides, dicumene peroxides, etc. commonly used in this type of polymerization reaction can be used, and the amount added is 0.1 to 10% by weight.
It is. The crosslinked polymer of the present invention can be produced by any polymerization method such as solution polymerization, bulk polymerization, suspension polymerization, etc., and the polymerization reaction is preferably carried out under an atmosphere of an inert gas such as nitrogen. ℃, preferably 60-80
The polymerization time is 3 to 20 hours. The diluent used in the suspension polymerization reaction is not particularly limited, but for example, the following can be used alone or as a mixture. i.e. aromatic hydrocarbons such as benzene, toluene, xylene, etc.
- Aliphatic hydrocarbons such as octane, alcohols such as cyclohexanol, lauryl alcohol, etc., and the amount added is 10 to 100% based on the monomer mixture.
%. Ordinary suspension polymerization of the monomer mixture, polymerization initiator, and diluent mixture can be carried out in a dispersion medium containing an appropriate protective colloid and salt.
That is, a suitable diluent such as water is added to a monomer mixture capable of introducing an inter-polymer complex structure into a crosslinked polymer, a crosslinking agent and a radical polymerization initiator, and cyclohexanol, which is immiscible with the monomers and diluent, is prepared. Polymerization can also be carried out in the presence of a suitable protective colloid or surfactant using liquid paraffin or the like as a dispersion medium. The crosslinked polymer obtained by the method described above is separated from the reaction solution by filtration, washed with a solvent such as pure water, methanol, or acetone, and then washed with an alkali, acid aqueous solution, or pure water. Wash repeatedly and dry under reduced pressure at 40-80°C. Adsorption and separation of substances using the crosslinked high molecular weight polymer of the present invention can be carried out by commonly used batch methods, column methods, etc., and they can be suitably selected as necessary to carry out the adsorption and separation. (g) Effects of the invention The crosslinked polymer containing the inter-polymer complex-forming component of the present invention can contain metal ions, basic amino acids,
It can adsorb and separate a wide range of substances such as antibiotics, proteins, enzymes, etc. It has particularly excellent adsorption ability for proteins and enzymes, and can be easily adsorbed and desorbed by adjusting the pH, making it easy to operate. It is simple, requires less equipment, and is an industrially useful adsorbent. (H) Field of Application of the Invention The crosslinked polymer obtained by the present invention is specifically a crosslinked polymer containing an interpolymer complex-forming component consisting of polyoxyethylene, polymethacrylic acid, polyacrylic acid, etc. It is a grafted type of POE chain, and since one end of the POE chain is free, the mobility of the POE chain due to the formation of a complex between polymers and dissociation is remarkable, making it effective for adsorption and separation of substances, making it useful as an adsorption carrier. It is also a highly practical compound that can be applied to immobilized enzyme carriers, etc. To further explain the range of application, this crosslinked polymer is inherently a weakly acidic cation exchange resin, and can be used to separate substances that are related to general weakly acidic cation exchange resins, such as calcium ions (Ca ⁺⁺ ), magnesium ions, etc. (Mg ⁺⁺ ), basic amino acids such as arginine, histidine, lysine, ornithine, antibiotics such as streptomycin, penicillin, chloramphenicol, and chlorotetracycline, and basic dyes such as methylene blue. It is possible to apply both adsorption and separation. In addition, this cross-linked polymer contains polyoxyethylene chains, which are synthetic polymers that only have mild interactions with biological components, and this
It exhibits adsorption and desorption properties through changes due to pH, and it exhibits adsorption and desorption properties through changes due to pH.
Suitable for use in separating or adsorbing proteins such as lactalbumin, β-lactoglobulin, chymotrypsinogen, γ-chymotrypsin, and human leukocyte interferon, enzymes, and biological substances such as red blood cells, granulocytes, and lymphocytes. . (li) Specific Examples of the Invention The present invention will be specifically explained in the following Examples, but the present invention is of course not limited to these Examples. Example 1 A 100 ml three-necked flask equipped with a stirring device and a cooling tube was charged with the following raw materials so that the molar ratio of acrylic acid and oxyethylene units was 1. That is, 10.44 g of acrylic acid, 3.02 g of ethylene glycol dimethacrylate, and methoxypolyoxyethylene methacrylate (oxyethylene units are 90).
Add 6.54g, azoisobutyronitrile 0.30g, and toluene 30g, and polymerize at 60℃ under nitrogen atmosphere.
Time polymerization was carried out. The obtained white bulk cross-linked polymer was filtered, purified water,
After washing with methanol and acetone, washing with alkali, acid aqueous solution and pure water, drying under reduced pressure at 80°C, and grinding in a mortar to obtain 17.88 g of white powder. The infrared absorption spectrum of this crosslinked polymer has absorption bands around 1100 cm ^-1 , 1710 cm ^-1 , and 3450 cm ^-1 , confirming the presence of carboxyl groups derived from acrylic acid and ether bonds derived from methoxypolyoxyethylene methacrylate. It was done. The percentage composition of this crosslinked high molecular weight polymer according to elemental analysis was as follows. C: 52.77 H: 6.85 The degree of crosslinking was 9.42 mol% of ethylene glycol dimethacrylate. The polymerization reaction proceeds as a matrix polymerization in which acrylic acid forms an intermolecular complex with polyoxyethylene chains while polymerizing, so acrylic acid blocks corresponding to polyoxyethylene chains are generated, and these blocks form ethylene glycol, which is a crosslinking agent. It is crosslinked with dimethacrylate, and from the above analysis results, this crosslinked polymer contains the following structure, Ratio of each block l:m:n=14500:161:
It is 1520. Note that this crosslinked high molecular weight polymer was insoluble in ordinary organic solvents. Example 2 A 500 ml decomposition flask equipped with a stirring device and a cooling tube was charged with the following raw materials so that the molar ratio of methacrylic acid and oxyethylene units was 1. Namely 21.25g of methacrylic acid, 5.15g of ethylene glycol dimethacrylate, methoxypolyoxyethylene methacrylate (oxyethylene unit is 9)
13.60 g of azoisobutyronitrile, 0.60 g of azoisobutyronitrile, 5.00 g of toluene mixed and dissolved, and 200 ml of a 1.3 g/ml aqueous calcium chloride solution containing 1% polyvinyl alcohol were added, under a nitrogen atmosphere, at a polymerization temperature of 70°C.
Suspension polymerization was carried out for 8 hours at a stirring speed of 300 rpm. The obtained porous white crosslinked polymer was filtered, washed with pure water, methanol, and acetone, and then treated with alkali,
It was washed with an acid aqueous solution and pure water, and dried under reduced pressure at 80°C.
36.24 g of white spherical powder with a diameter of about 100 μm was obtained. The infrared absorption spectrum of this crosslinked polymer has absorption bands around 1100 cm ^-1 , 1720 cm ^-1 , and 3450 cm ^-1 , confirming the presence of carboxyl groups derived from methacrylic acid and ether bonds derived from methoxypolyoxyethylene methacrylate. The percentage composition of this crosslinked polymer was determined by elemental analysis to be as follows. C; 55.67 H; 7.48 ¹³ CNMR spectrum was also measured as follows. A predetermined amount of heavy water was added as a magnetic field locking agent under predetermined PH conditions, and the crosslinked polymer was immersed in it to measure ^{the 13} CNMR spectrum at room temperature. 3000～
By integrating 30,000 times, the signal of the ether carbon derived from the oxyethylene chain in the crosslinked polymer is determined based on tetramethylsilane.
It was observed at around 70 ppm (this proves that ether carbon derived from methoxypolyoxyethylene methacrylate was introduced into the crosslinked polymer). Table 1 shows the relationship between the half-width of the ether carbon signal and the pH.

[Table] The degree of crosslinking was 8.65 mol% of ethylene glycol dimethacrylate. This crosslinked polymer also produced methacrylic acid blocks corresponding to polyoxyethylene chains for the same reason as described in Example 1, and these were crosslinked with ethylene glycol dimethacrylate as a crosslinking agent.
Combining the above analysis results, this crosslinked polymer contains the following structure, Ratio of each block l:m:n=2470:274:260
It is. Note that this crosslinked high molecular weight polymer was insoluble in ordinary organic solvents. Furthermore, as shown in Table 1, it can be seen that by changing the pH, the characteristics of the crosslinked polymer, that is, the mobility of the polyoxyethylene chains in the present crosslinked polymer, change greatly. Next, the adsorption capacity of bovine serum albumin (BSA) was measured using this crosslinked polymer. That is, 1g of crosslinked polymer and 50mg/dl of the specified pH
After reaching the equilibrium adsorption amount of 20ml of BSA solution at room temperature,
The amount of BSA in the supernatant was determined by the microbiuret method. The results are shown in Table 2.

[Table] As shown in Table 2, it can be seen that the adsorption characteristics of crosslinked polymers change greatly by changing the pH. The reason why this cross-linked polymer showed significant adsorption ability at pH 4.9 is that the carboxyl group does not dissociate in the cross-linked polymer, and moreover, it forms a stable inter-polymer complex with the polyoxyethylene chain. And moreover
This is because the cross-linked polymer was stably adsorbed mainly due to hydrophobic interaction because the BSA molecule has no charge and has a compact molecular shape because it is near the isoelectric point of the BSA molecule. it is conceivable that.
In any case, as is clear from Table 2, the crosslinked polymer of the present invention can be
It can easily adsorb and desorb BSA, making it an excellent adsorbent. Therefore, by using this crosslinked polymer, the target substance can be easily adsorbed and separated by simply adjusting the pH, and it is found that it is a very useful polymer adsorbent from an industrial perspective. Example 3 A 500 ml decomposition flask equipped with a stirring device and a cooling tube was charged with the following raw materials so that the molar ratio of methacrylic acid and oxyethylene units was 1. Namely 22.65g of methacrylic acid, 5.49g of ethylene glycol dimethacrylate, methoxypolyoxyethylene methacrylate (oxyethylene unit is 90)
Add 200 ml of a 1.3 g/ml calcium chloride aqueous solution containing 11.87 g, 75% benzoyl peroxide, 1.20 g, and toluene 4.00 g, and 1% polyvinyl alcohol, under a nitrogen atmosphere, at a polymerization temperature of 70°C.
Suspension polymerization was carried out for 8 hours at a stirring speed of 300 rpm. The obtained porous white crosslinked polymer was filtered, washed with pure water, methanol, and acetone, and then treated with an alkali,
It was washed with an acid aqueous solution and pure water, and dried under reduced pressure at 80°C.
34.08 g of white spherical powder with a diameter of about 100 μm was obtained. The infrared absorption spectrum of this crosslinked polymer has absorption bands around 1100 cm ^-1 , 1710 cm ^-1 , and 3450 cm ^-1 , confirming the presence of carboxyl groups derived from methacrylic acid and ether bonds derived from methoxypolyoxyethylene methacrylate. It was done. The percentage composition of this crosslinked high molecular weight polymer according to elemental analysis was as follows. C: 55.79 H: 7.52 In addition, ^{the 13} CNMR spectrum was measured in the same manner as in Example 2. In this crosslinked polymer, a signal of ether carbon derived from oxyethylene chains was observed at around 70 ppm based on tetramethylsilane. This shows that ether carbon derived from methoxypolyoxyethylene methacrylate has been introduced into the crosslinked polymer. Next, the half-width of this ether carbon signal and
The relationship with PH is shown in Table 3.

[Table] The degree of crosslinking was 9.43 mol% of ethylene glycol dimethacrylate. This crosslinked polymer also produces methacrylic acid blocks corresponding to polyoxyethylene chains for the same reason as described in Example 1, and these are crosslinked with the crosslinking agent ethylene glycol dimethacrylate. Combining the above analysis results, this crosslinked polymer contains the following structure, Ratio of each block l:m:n=26300:292:
It is 2770. Note that this crosslinked high molecular weight polymer was insoluble in ordinary organic solvents. Furthermore, as shown in Table 3, it can be seen that by changing the pH, the properties of the crosslinked polymer, that is, the mobility of the polyoxyethylene chains in this crosslinked polymer, change greatly. Moreover, this rate of change is larger than the change shown in Example 2. This shows that a larger difference in adsorption characteristics can be expressed by changing the pH, and shows that it is effective in separating and adsorbing substances. Next, the BSA adsorption capacity was measured in the same manner as in Example 2 using this crosslinked polymer. The results are shown in Table 4.

[Table] As shown in Table 4, it can be seen that the adsorption characteristics of crosslinked polymers change greatly by changing the pH. Further, the adsorption behavior can be understood for the same reason as described in Example 2. As is clear from Table 4, the crosslinked polymer of the present invention can easily adsorb and desorb the substance to be separated by changing the pH. It can be seen that the target substance can be easily adsorbed and separated simply by adjusting the pH, making it a useful polymeric adsorbent. In addition to BSA, similar separation and adsorption of proteins such as α-lactalbumin and chymotrypsinogen, enzymes, red blood cells, granulocytes, lymphocytes, etc., could be carried out using this crosslinked polymer. Example 4 50 mg of the same crosslinked polymer as in Example 1 was soaked in physiological saline in advance to sufficiently swell it, and then excess physiological saline was removed. Add this to 2ml of human plasma.
was added and stirred at 37°C for 2 hours to allow adsorption, then the supernatant was taken out and the adsorption rate was determined by quantifying the amount of residual protein. Albumin, IgG (nomnoglobulin G), IgM
(Immunoglobulin M) is RID method (immunodiffusion method)
The immune complex was quantified by the PEG (polyethylene glycol) precipitation method-RID method. The results are shown in Table 5.

[Table] It can be seen that in addition to albumin, proteins such as IgG, IgM, and IC are effectively adsorbed by the adsorbent of the present invention. In this example, the pH was approximately 7.0.
The reason why the amount of albumin adsorbed is small is thought to be due to the influence of this pH.

Claims (1)

[Claims] 1 General formula (), (In the formula, each R may be the same or different and represents H or an organic group, and l, m, and p are integers of 1 or more.) A crosslinked polymer containing an interpolymer complex-forming component represented by Adsorbent made of polymer. 2 A: A compound having a polyoxyethylene chain and a polymerizable double bond at the same time in the molecule; B: A carboxylic acid or its derivative having a polymerizable double bond; C: A general compound characterized by reacting with a crosslinking agent. formula(), (In the formula, each R may be the same or different and represents H or an organic group, and l, m, and p are integers of 1 or more.) A crosslinked polymer containing an interpolymer complex-forming component represented by A method for producing an adsorbent made of a polymer.