CS260852B1 - Macroporous polymer sorbents and method of their production - Google Patents

Description

The invention relates to macroporous cupolymer sorbents derivatized on the inner pore surface, immobilizing functional group-containing polymers, irreversibly penetrating them into the pore surface of the macroporous copolymers, or further stabilizing by covalent bonding of these polymers to the primary matrix of the copolymer support.

Organic polymeric sorbents for separation purposes are finding increasingly widespread applications. The development of instrumentation increases the demands on the quality of the analytical columns in liquid chromatography, especially the requirements for higher pressure resistance, good flow and, last but not least, chemical and microbiological stability are increasing.

Macroporous copolymer sorbents are widely used in the technology field for the separation of complex compound mixtures. In particular, natural systems containing proteins, glycoproteins, nucleic acids, polysaccharides, lipids and others are increasingly subject to intensive use in which separation techniques and technologies play a paramount role. In addition to the required properties of analytical sorbents, easy regenerability, high reproducibility of chemical and physical parameters, and the absence of undesirable non-specific and undefined interactions are required.

Sorption materials for ion exchange chromatography, affinity chromatography, gel permeation chromatography, and reverse phase chromatography employing macroporous organic polymers as carriers have developed considerably, since they are largely more chemical resistant than inorganic sorbents. A certain disadvantage in comparison with, for example, silica-based sorbents is their lower pressure stability, varying degrees of swelling depending on the pH, the ionic strength of the medium and the type of liquid phase with which they are in contact. The higher content of crosslinking agents naturally increases the mesh density and thus the mechanical resistance of the copolymer, but also the final physical properties of the heterogeneous three-dimensional copolymers are influenced and also limited by the presence of the dispersed phase in the polymerization. The effects of porogens, the flexibility of the primary polymeric chain constituting the network, intermolecular interactions in polymerization, etc. must be taken into account. Considerations and practical experiments have resulted in mechanical and physico-chemical constraints on the mechanical sta260852

280852 bility, which cannot be exceeded in single-stage copolymerization.

If the crosslinker content increases when copolymerizing the monovinyl monomer with a divinyl or polyvinyl crosslinker, the content of unreacted " hanging " double bonds remains in the final copolymers [J. ŠTOKR, B. SCHNEIDER, A. FRYDRYCHOVÁ and J. COUPEK, J. Appl. Polymer Sci. 23 (1979) 3,553-3,561]. If it is 0 ¹ residual vinyl divinylbenzene after reaction a first vinyl reactivity of the remaining vinyl groups somewhat reduced. In contrast, the reactivity of the residual vinyl of ethylene dimethacrylate is not limited by resonant effects and remains unchanged even after the reaction of the first vinyl group of this monomer. The presence of residual vinyl in the macroporous matrix can be advantageously used to further copolymerize it with other monomers and to immobilize the entire polymer chain which is formed by covalent bonding to the original macroporous matrix during polymerization.

The present invention provides macroporous polymeric sorbents comprising a primary macroporous carrier which is a copolymer of at least one monovinyl monomer selected from the group consisting of 2-hydroxyalkylmethacrylates of 2 to 4 carbon atoms in alkyl, alkylmethacrylates of 1 to 4 carbon atoms in alkyl , methacrylamide, C1 -C4 alkyl acrylates and styrene with at least one polyvinyl monomer selected from C2 -C4 alkylene dimethacrylates, esters of polyhydric alcohols with at least two methacrylic acid residues and divinylbenzene or divinyl homopolymer or a polyvinyl monomer selected from the group consisting of esters of acrylic or methacrylic acid with divalent or polyhydric alcohols and divinylbenzene in which an acrylic polymer carrying functional groups selected from the boron comprising vinyl-, allyl-, epoxy-, nitrile-, amido-, carbo-, amino-, alkyl-, aryl-, groups and derivatives thereof.

Irreversible immobilization can be achieved, for example, by physical penetration - the ingrowth of the resulting polymer with the primary three-dimensional matrix of the matrix. Copolymerization of the hanging vinyl groups of the primary matrix with the polymer formed is preferred. Radical polymerization in the presence of initiator is achieved by raising the temperature to 60-110 degrees Celsius, thermal polymerization in the absence of initiator is achieved by raising the temperature to 100-150 ° C. The free-radical polymerization can also be initiated by radiation of a suitable wavelength. In some cases, it is preferable to evaporate the solvent prior to polymerization. For chromatographic and other applications, spherical sorbents are preferred.

The sorbents of the present invention are prepared by contacting a primary macroporous carrier with a monomer mixture consisting of a solution of acrylic monomer bearing functional groups and optionally a polymerization initiator in a solvent, and monomer mixture sorbed on the inner surface of the primary macroporous carrier for polymerization.

The choice of monomer to be used to immobilize the resulting polymer on the primary macroporous matrix depends on the effect to be achieved. When the monomer is a divinyl or polyvinyl compound, the polymerization and copolymerization with the hanging residual double bonds of the primary matrix results in subsequent crosslinking, which increases the network density and further reduces its swelling and mechanical stability of the end product. This effect is further enhanced by the penetration of the monomer into the surface layers of the primary matrix and its ingrowth by the newly formed polymer. The resulting effect of subsequent crosslinking on the pore size distribution and the specific surface area value is not great (see examples), the mechanical properties of such derivatized sorbents are significantly improved. If the monomer contains a functional group such as hydroxyl (2-hydroxyethyl methacrylate) or an amide (methacrylamide) that is non-ionic and hydrophilic, then the polymer-derivatized sorbent formed exhibits significantly increased surface hydrophilicity and a reduction in non-specific hydrophobic adsorption.

It is therefore suitable for rapid gel chromatography of biopolymers. When the functional group of the monomer is alkyl or aryl (methyl methacrylate, butyl methacrylate, methacrylic acid anilide, etc.), the end product after immobilization of the resulting polymer on the inner surface of the three-dimensional matrix acquires hydrophobic properties which can be controlled and advantageously used for adsorption (hydrophobic) chromatography of low molecular weight biopolymers.

Many vinyl monomers contain ionic functional groups (methacrylic acid, acrylic acid, dimethylaminomethyl acrylate, diethylaminoacrylate, etc.). By immobilizing the polymers of these ionic monomers, the product acquires the properties of macroporous ion exchangers which, depending on the nature of the functional groups, are strong or weak anion exchangers or cation exchangers for use in ion chromatography and ion exchange chromatography.

Monomers containing chemically reactive groups (glycidyl methacrylate, p-nitrophenyl methacrylate ester, etc.) have also been used, and immobilized polymers in this case provide a reactive sorbent with a very wide range of subsequent reactions utilizing covalent bonding of both low and high molecular weight ligands.

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The main fields of application of these sorbents with high selectivity and specificity are bioaffinity chromatography, ligand exchange chromatography and chromatofocusing.

The inert solvent used also plays an important role in the derivatization of the present invention. Its solvation potential for the primary polymer matrix determines the degree and depth of swelling of the surface layer of the primary matrix and the depth of penetration of monomer and initiator into the surface layer. Generally, liquids that are a precipitant for the primary matrix (e.g., heptane pro, 2-hydroxyethyl methacrylate co-ethylene dimethacrylate copolymer) ensure immobilization of the polymer only in the thin surface layer of the primary matrix or only at the interface.

Naturally, the solvating ability of the monomer itself for the primary matrix also plays a role (especially in solvent-evaporating processes) and mixed interactions in the primary matrix-solvent-monomer system, which are determined by the concentration ratios of all three components. Also, the mechanism of polyinerization on the surface of the primary matrix affects the physical and physicochemical properties of the resulting product.

From the foregoing, it is obvious that the present invention also includes the possibility of binary and ternary or multiple copolymerization with the immobilization of copolymers on the surface of the primary macroporous matrix, and thus an extremely wide range of properties to control the properties of the resulting sorbents.

For most chromatographic and other applications it is desirable to use sorbents whose particles have a spherical shape. It is therefore advantageous to select copolymers prepared by suspension polymerization as primary sorbents with a macroporous matrix, which are the subject of a number of protected processes. If a large number of binding sites are required for the covalent immobilization of the polymer on the surface of the matrix, it is desirable to select primary sorbents with a high crosslinker concentration or reaction products with incomplete conversion of the crosslinker so as to guarantee higher content of hanging vinyl functional groups in the primary matrix. The invention is further illustrated and exemplified by a number of examples which are not to be construed as limiting.

In some examples, monomers, initiators, and primary copolymer macroporous carriers are designated by trade names to shorten and provide clarity. The physical and chemical characteristics of these materials are as follows:

SEPARON HEMA 300 - macroporous copolymer of 2-hydroxyethyl methacrylate with ethylenedimethacrylate with a molecular weight exclusion limit of 300,000 daltons and a specific surface area of 60 m ² / g

SEPARON HEMA 1,000 - · Macroporous copolymer of 2-hydroxyethyl methacrylate with ethylene dimethacrylate with a molecular weight exclusion limit of 2.10 ⁶ Daltons and a specific surface area of 50 m ^? /G

SEPARON SE - macroporous copolymer of styrene with ethylene dimethacrylate with a specific surface area of 70 m ² / g and a content of styrene and divinylbenzene in 1: 1 copolymers

SEPARON BD - macroporous copolymer of butyl methacrylate with divinyl benzene with specific surface 120 m- / g

HEMA-2-hydroxyethyl methacrylate, p. A, dest.

ABIBN - azobisisobutyronitril kryst. p. a. EDMAM - ethylenedimethacrylate

Example 1

To a suspension of 10 g Separon HEMA 300 (30-40 gmj in 50 ml of scarred acetone) was added a residue of 1 g of HEMA in 5 ml of distilled acetone and 1 ml of a 1% solution of ABIBN in distilled acetone. The sorbent treated, containing a uniformly loaded monomer with the initiator, was then dry heated in a flask to 70-110 [deg.] C. After washing with water, methanol, acetone and ether, a sorbent was obtained whose swelling in water was reduced by 33%, in methanol by 23% and in acetone by 20%.

Example 2

Application of the HEMA monomer and the initiator to HEMA Separon 300 (30-40 _(umj was performed in the same amounts and the same procedure as described in Example 1. The modified sorbent was then converted into n-heptane and heated therein with stirring for 7 h to reflux temperature (90-95 ° C). the product was further treated as in Example 1, specific surface area increased by 25 ° / o, the value of an exclusion limit remained unchanged ^300.10:! D.

Example 3 g Separone HEMA 200 (30-40 μπι) was added to 50 ml n-heptane, stirred for 30 minutes at room temperature and then a mixture of 1 g HEMA and 1 ml 1% acetone solution ABIBN was added dropwise with stirring over 15 minutes. The reaction mixture was then stirred for an additional 30 minutes at room temperature and then heated to reflux for a minimum of 7 hours. The product was transferred for filtration, washed with acetone, water, methanol and ether, extracted with benzene, ethanol and acetone every 8 hours and dried.

Example 4

Separon HEMA 1000 (12 μΐη) was modified with the same weight and procedure as described in Example 3. The product treatment was also carried out according to Example 3. The exclusion limit was the same as that of the starting Separon, i.e. 2.10 ⁶ D, the specific surface area increased. 17% (from 46.5 to 54.3 m ² / g).

Example 5 5 g Separone HEMA 1000 (12 µm) was added to 50 ml n-heptane and 1 g HEMA and 1 ml 1% acetone ABIBN solution were added to this suspension. The mixture was stored overnight at 5-10 ° C and the next day polymerized by heating to 95 ° C for 7 hours. Treatment of the product of Example 3.

Example 6 g Separone HEMA 1000 (12 µm) was doped with 1.5 g HEMA with 1 ml 1% acetone solution ABIBN in 50 ml n-heptane as described in Example 3.

Example 7 g Separone HEMA 1000 (12 μΐη) 250 milliliters i-octane, 3.5 g HEMA, 1.5 g EDMA, 5 ml 1% acetone solution ABIBN were polymerized as described in Example 3.

Example 1 a d 8 g Separone HEMA 1000 (12 μπι), 250 ml i-octane, 3.5 g HEMA, 1.5 EDMA, 5 ml 1% acetone solution ABIBN were polymerized as described in Example 3.

Example 9 g Separone HEMA 1000 (15-20 / ml) 250 ml n-heptane, 4.5 g HEMA, 3 g EDMA and 0.075 ml 10% acetone solution ABIBN were polymerized as described in Example 3.

Example 10 g Separone HEMA 1000 (15-25 µm) 250 ml n-heptane, 1.5 g HEMA, 1 g EDMA, 0.025 ml 10% ABIBN solution were polymerized as described in Example 3. Example 11 g Separone HEMA 1 000 (15-25 μπι) 250 ml n-heptane, 15 g glycidyl methacrylate, 1.5 ml 10% acetone solution ABIBN was polymerized according to example

3. Transfer the product with acetone for filtration, wash 3 times quickly with water, then with acetone and ether. Finally, the product was extracted for 8 hours with benzene and 8 hours with acetone. 55.5 g of sorbent having an epoxy content of 1040, ueq / g) were obtained.

Example 12 g Separone HEMA 1000 (15-20 μπι) 150 ml n-heptane, 6 g glycidyl methacrylate, 0.6 ml 10% solution of ABIBN in acetone were polymerized and the product treated as described in Example 11. The product obtained (32.25) gj contained 736 / epoxy groups / g sorbent.

EXAMPLE 13 g of Separone HEMA 1000 (10 μΐη), 100 ml of n-heptane, 3 g of glycidyl methacrylate and 3 ml of a 1% acetone solution of ABIBN were polymerized according to Example 11. The product contained 682 .mu.g / g of epoxy groups.

Example 14

31.3 g of the sorbent prepared according to Example 12, 155 ml of n-heptane, 6.2 g of glycidyl methacrylate and 0.6 ml of a 10% acetone solution of ABIBN were polymerized and worked up as described in Example 12. A product containing 1250 <eq of epoxy groups was obtained / g sorbent.

Example 15 g of the sorbent prepared according to example 14, 75 ml of n-heptane, 3 g of glycidyl methacrylate and 0.3 ml of a 10% solution of ABIBN in distilled acetone were polymerized and processed as in Example 12. A sorbent with an epoxy group content of 1680 juq / g was prepared. .

Example 16 g Separone HEMA 1000 (15-25 μΐη), 250 ml n-heptane, 6 g glycidyl methacrylate, 4 g EDMA, 1 ml 10% ABIBN in acetone were polymerized and treated as in Example 12. 57.6 g of product were obtained. content of epoxy groups 460 / zekv / g sorbent.

g of the thus obtained ¹ sorbent containing epoxy groups was suspended in 250 ml of 0.1 M perchloric acid and allowed to react at room temperature for 5 days. The sorbent was then filtered, washed with distilled water to neutralize the filtrate, followed by methanol, acetone and ether.

Example 17 g of the product of Example 16 was polymerized in 100 ml of n-heptane with 4 g of glycidyl methacrylate and 0.4 ml of a 10% acetone260852 solution of ABIBN as described in Example 11. Sorbent thus prepared. contained 690 epoxy groups / g sorbent. Example 18 g Separone HEMA 1000 (30-45 µm), 150 ml n-heptane, 4.5 g acrylonitrile and 0.45 ml 10% solution of ABIBN in dist. Acetone was polymerized as described in Example 3. The product obtained contained 1,643 µg CN groups / g sorbent.

EXAMPLE 1 9 g Separone HEMA 1000 (30-45 μιη), 250 ml n-heptane, 7.5 g acrylamide dissolved in 35 ml dist. acetone and 0.75 ml of a 10% solution of ABIBN in the rain, the acetone was heated to boiling. By distilling off the required amount of acetone, the composition of the reaction mixture was adjusted so that the boiling point was at least 80 ° C and maintained at this temperature for 7.5 hours. The product was worked up as described in Example 3. The amide groups content of the product was 1670 juq / g. sorbent.

EXAMPLE 20 0 g Separone HEMA 1000 (30-45 μηι), 250 ml n-heptane, 7.5 g acrylamide dissolved in 35 ml dist. of acetone, 5 g of EDMA, 1.25 ml of 10% acetone solution of ABIBN was polymerized as described in Example 19. The amide group content of the product was 1671 µg / g sorbent.

Example 21 g Separone HEMA 1000 treated according to Example 13 was suspended in 75 ml diethylamine and 2 ml water and heated to boiling for 8 h. The sorbent was then filtered, washed with dest. water to neutralize the filtrate, then washed with methanol and ether. The sorbent thus prepared had an ion exchange capacity of 564 µg / g sorbent, an N content of 1.09%.

Example 22

4.7 g of Separone HEMA 1000 treated according to Example 13 were suspended in 50 ml of triethylamine and 1.8 ml of water and heated to reflux for 20 hours. The product after treatment according to Example 21 contained 0.68% N.

Example 23 g Separone SE (15-25 .mu.m) was reacted without initiator with 1.5 g divinylbenzene according to Example 1. The resulting product was heated to 150 ° C for 24 hours after evaporation of the solvent.

The resulting crosslinked product was used to absorb organic compounds from aqueous solutions.

EXAMPLE 24 g of macroporous copolymers of butyl acrylate with trimethylolpropane trimethacrylate having an elimination limit of 200,000 daltons and a specific surface area of 105 m ² / g were reacted with 7 g of dimethylaminoethyl methacrylate analogously to Example 1. The resulting product had an ion exchange capacity of 900 µg / g.

Example 25

To 10 g Separone BD (100-200 µm) in 50 ml distilled acetone was added a solution of 1 g acrylic acid and the suspension was evaporated to dryness on a rotary evaporator below 30 ° S. The treated sorbent is placed in the vicinity of the hard radiation source for 48 hours. The resulting product, after extraction, was analyzed for carboxyl group content (120 µeq / g). Example 26 g of a macroporous homopolymer of ethylene dimethacrylate with an inner surface area of 300 m ² / g was derivatized analogously to Example 11 with methacrylic acid anilide. The resulting product was diazotized and used to immobilize naphthalenesulfone compounds.

EXAMPLE 27 g of a macroporous homopolymer of divinylbenzene having an internal surface area of 270 m @ ² / g were processed analogously to Example Ϊ except that a mixture of 1.5 g of diallyl phthalate and 1.5 g of acrylonitrile was adsorbed on the support. The reaction conditions were the same as in Example 1. The resulting product contained 1090 juevv nitrile groups / g.

Example 28 g Separone HEMA 1000 (10 μΐη), 50 ml n-heptane, 1.5 g Ν, Ν-methylene-bis-acrylamide in 7 ml dist. of acetone, 0.15 ml of a 10% acetone solution of ABIBN was polymerized as described in Example 19. The amide group content of the product was 1 607 µg / g sorbent.

Example 29 g Separone HEMA 1000 (10 µm), 50 ml n-heptane, 1.5 g triethylene glycol methacrylate, 0.15 ml 10% acetone solution ABIBN was polymerized as described in Example 19.

EXAMPLE 30 g of Separone HEMA 1000 (10 .mu.m), 50 ml of n-heptane, 1.5 g of methacrylic acid, 0.15 ml of a 10% acetone solution of ABIBN were polymerized as described in Example 19. The product contained 573, COOH / g. sorbent.

EXAMPLE 31 g of a trimethylolpropantrimethacrylate macroporous homopolymer with an exclusion limit of 10 ⁶ daltons in a heptane solution were derivatized analogously to Example 3, except that 1.5 g of dimethylaminoethyl methacrylate was used as monomer. The resulting ion-exchange capacity determined by titration was 950 ueq / g dry sorbent.

Example 32 g of a macroporous homopolymer of divinylbenzene having a specific surface area of 400 m ² / g was derivatized analogously to Example 1 with a solution of 1 g of methacrylic acid in 5 ml of acetone with the addition of 1 ml of a 1% acetone solution of benzoyl peroxide. The resulting product after titration showed an exchange capacity of immobilized carboxyl groups of 950 µg / g sorbent.

Claims (9)

fRedmEt CLAIMS 1. Macroporous polymeric sorbents, characterized in that they consist of a primary macroporous carrier, which is a copolymer of at least one monovinyl monomer selected from the group consisting of 2-hydroxyalkyl methacrylates of 2 to 4 carbon atoms in alkyl, alkyl methacrylates of 1 to 4 carbon atoms in alkyl, methacrylamide, C 1 -C 4 alkyl acrylates and styrene with at least one polyvinyl monomer selected from the group consisting of C 2 -C 4 alkylene dimethacrylates, esters of polyhydric alcohols with at least two methacrylic acid and divinylbenzene residues or a divinyl or polyvinyl monomer homopolymer selected from the group consisting of an ester of acrylic or methacrylic acid with divalent or polyhydric alcohols and divinylbenzene, in which an acrylic polymer bearing functional groups selected from vinyl-, reversibly immobilized on the inner surface of the pores, alkyl-, epoxy-, nitrile-, amido-, carboxy-, amino-, alkyl-, aryl-, groups and derivatives thereof. 2. Sorbents according to claim 1, characterized in that the polymers irreversibly retained on the surface of the carrier are immobilized by physically penetrating the polymer into the three-dimensional network of the carrier. 3. Sorbents according to claim 1, characterized in that: INVENTION that polymers irreversibly retained on the surface of the carrier are immobilized by covalent bonding of the polymer to the three-dimensional matrix of the carrier. 4. Sorbents according to claim 1, characterized in that their particles have a spherical shape. 5. A process for the production of macroporous polymeric sorbents according to claim 1, characterized in that the primary macroporous carrier as defined in item 1 is contacted with a monomer mixture consisting of a solution of acrylic monomer carrying functional groups as defined in item 1 and optionally a polymerization initiator in solvent and monomer mixture. sorbed on the inner surface of the primary macroporous carrier is brought to polymerization. 6. A process according to claim 5, wherein the radical polymerization of the woolly monomer is carried out on the support surface by raising the system temperature to 60-110 [deg.] C. 7. A process according to claim 5, characterized in that the polymerization of the vinyl monomer is carried out thermally on the support surface in the absence of an initiator by raising the temperature to 100-150 ° C. 8. A process according to claim 5, wherein the polymerization of the vinyl monomer is carried out on the surface of the support by radiation. 9. The process of claim 5 wherein the solvent is evaporated prior to polymerization.