CN104387712A - Nano composite carrier with superparamagnetism and preparation method thereof - Google Patents
Nano composite carrier with superparamagnetism and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a nano composite carrier with superparamagnetism and a preparation method thereof. The nano composite material takes nano Fe3O4 as a core and polyacrylamide gel (PAAG) as a shell, combines advantages of nano Fe3O4 and polyacrylamide gel, is a very ideal immobilized enzyme carrier, and can also be used as a carrier for immobilized cells, slow-release drugs and targeted drugs. The preparation method comprises the following steps: (1) preparing a mixed solution of acrylamide, methylene diacrylamide and nano Fe3O4; (2) preparing a reversed-phase micro-emulsion by taking the mixed solution as a disperse phase; (3) adding a catalyst and an accelerant, which are used for generating polyacrylamide gel, into the reversed-phase micro-emulsion to obtain a composite material; and (4) separating and purifying to obtain a nano Fe3O4/PAAG composite material. The nano composite material provided by the invention js capable of shortening the recovery time of enzyme and improving the recovery efficiency when serving as an enzyme carrier, and meanwhile, the preparation condition is mild, the process is simple and the cost is low.
Description
Technical field
The invention belongs to technical field of nano material, be specifically related to nano combined carrier with superparamagnetism and preparation method thereof.
Background technology
Enzyme is the biomacromolecule with catalysis, it is the catalyzer of all metabolic reactions in organism, it is the base substance maintaining organism vital movement, have catalytic reaction condition gentleness, catalytic efficiency is high, and Substratspezifitaet is strong, by product is few, applied range, the advantages such as catalytic activity is easy to adjust are a kind of desirable catalyzer.Along with the development of the technology such as biotechnology, enzyme engineering, molecular biology, zymin all obtains in fields such as molecular diagnosis, diseases prevention and treatment, medicine preparation, agriculture production, food-processing, environment protection and green chemical synthesis and develops rapidly and widespread use.Develop enzyme resource, promote enzyme suitability for industrialized production and application, realize the efficiency utilization of enzyme, to the development promoting China's medical and health cause, ensure the supply of the living matters such as food, improve Green Chemistry level of industrial technology, improve the ecological environment, realize sound and rapid economic growth, promote international competitiveness significant.
But the preparation process of enzyme is complicated, and reaction conditions requires harsh, and production cost is high.Simultaneously, its essence is by amino acid constitutive protein matter, space structure is unstable, very responsive to environment, the existence of the factors such as temperature, pH value, pressure, trypsinase, organic solvent, metal ion, oxygenant, reductive agent, electromagnetic field all likely makes its loss of catalytic activity, preserves more difficult.In addition, in catalytic reaction process, enzyme liquid is dispersed in reaction system, reclaim purification difficult, can not reuse, not only cause the waste of enzyme but also increase the purification of product and refining cost, and with the prolongation in reaction times, catalytic activity declines gradually, causes speed of response to reduce.Expensive, space structure is unstable, easy loss of activity, and enzyme liquid reclaims the bottleneck that difficulty etc. is considered to the use of restriction enzyme large-scale promotion.Therefore, on the basis of the space structure and catalytic performance that do not change enzyme, exploitation one can enhanced stability, improve catalytic activity, be convenient to again recycle, increase and recycle number of times, reducing the immobilization technology of use cost, is one of important research direction of enzyme engineering in recent years.
Utilize various organic/inorganic porous material, by physics or chemical process, enzyme load is prepared immobilized enzyme on the internal void or surface group of porous material, the organic efficiency of enzyme can be improved, increase cycle index, reducing production cost, is one of effective way promoting enzyme suitability for industrialized production and application.Fixation support affects enzyme after the immobilization of enzyme success and immobilization to live one of rate of recovery height, catalytic activity is strong and weak, cycle index is how many key factor, and the solid support material of exploitation excellent property is one of important development direction of immobilized enzyme.
Polyacrylamide gel (Polyacrylamide Gel, be called for short PAAG) be mixed by a certain percentage by acrylamide monomer and methylene diacrylamide linking agent, the cross network structure be polymerized under catalyst action, it can produce molecular sieve effect, is commonly used in the field such as chromatography media, electrophoretic separation support material.It has the advantages such as good biocompatibility, specific surface area is large, carrying capacity is strong, pore size is controlled.At present, the research that some patents and bibliographical information take PAAG as fixed enzyme vector is had.The patent of having applied for has: " preparation method of large empty polyacrylamide resin and application thereof " (patent No.: the ZL 201010244212.6) of the application such as Lei Zhongli; " a kind of method of fixing Formaldehyde decomposition pathway key enzyme and cofactor and application thereof " (patent No.: the ZL 201010224713.8) of the application such as Chen Limei.The paper delivered has: Tang Yu, etc. polyacrylamide gel immobilized enzyme method prepares the research of maltotetrose. Agriculture in Jiangxi journal, 2013,25 (7): 88-90; Liu Guiping, etc. the research of modified polyacrylamide immobilization Phenol-degrading Bacteria Strains. environmental engineering journal, 2008.6 (2): 861-864; Xue Ping, etc. polymer carrier is to the immobilization role of penicillin acylase. Journal of Functional Polymers, 2005,18 (2): 340-345; Li Yuanxun, etc. polyacrylic acid carrier is used for the immobilization of penicillin acylase. Journal of Functional Polymers, 2001,14 (4): 442-444.
But the method that these patents and document adopt is all prepare PAAG by polyreaction in the solution, then adopt the modes such as absorption to be fixed on by enzyme in the inner and surperficial hole of gel, it is even larger that the particle diameter as the PAAG of carrier is in micron order.In addition, also find no PAAG in prior art and be combined the report forming complex carrier with Z 250, more do not use PAAG to wrap the report being formed nano combined carrier by nano ferriferrous oxide.
In addition, use magnetic nano ferroferric oxide is core, core wraps and is also had report in the prior art, as the nano combined carrier being shell structure with chitosan, sodium alginate or alginate calcium etc. by the nano combined carrier of shell structure.But the diversity of object " immobilized enzyme " structure in view of the load of carrier institute, this area also needs the nano combined carrier kind developing other abundant species.
Summary of the invention
Present invention incorporates nanometer Fe
3o
4with the advantage of nano polypropylene acrylamide gel, invent one with nanometer Fe
3o
4for core, polyacrylamide gel is the nanometer Fe of shell
3o
4/ PAAG matrix material and preparation method thereof.
Compared with plain polypropylene acrylamide gel, nano polypropylene acrylamide gel has the advantages such as particle diameter is little, particle monodispersity is good, specific surface area large, even aperture distribution, carrying capacity are strong.Nanometer Fe simultaneously
3o
4there is the feature such as superparamagnetism and magnetic response Ying Qiang.The present invention proposes first with nanometer Fe
3o
4for core, polyacrylamide gel is the nanometer Fe of shell
3o
4/ PAAG matrix material and preparation method thereof, this material both can give full play to the high advantage of strong, the fixing rate of nano polypropylene acrylamide gel carrying capacity, can well utilize nanometer Fe again
3o
4superparamagnetism and magnetic response should be strong feature.With the immobilized enzyme of this material prepared by carrier, in catalytic reaction process or after reaction terminates, under the driving of magnetic field force, to a direction displacement, and can separate from reaction system, the recovery being convenient to immobilized enzyme with reuse.Therefore, nanometer Fe
3o
4/ PAAG matrix material is a kind of ideal fixed enzyme vector.
The present invention is that fixed enzyme vector and preparation thereof provide a kind of new thoughts and methods, likely become the breakthrough point promoting enzyme suitability for industrialized production and application, in enzyme engineering and biocatalysis field, there is huge using value, for raising China enzyme engineering technology, the development of the industry such as promote green chemical synthesis, biocatalysis and chiral drug to prepare is also significant.
Therefore, the invention provides a kind of nano combined carrier with superparamagnetism, described carrier is with nanometer Fe
3o
4be shell for core, with polyacrylamide gel.
In a kind of embodiment, the described nanometer Fe as core
3o
4particle diameter be 5 ~ 1000nm, be preferably 10 ~ 200nm, be more preferably 20 ~ 50nm; The described polyacrylamide gel thickness as shell is 5 ~ 1000nm, is preferably 10 ~ 800nm, is more preferably 30 ~ 500nm.Preferably, the particle diameter of described carrier is 15 ~ 1000nm, preferably 25 ~ 800nm, more preferably 50 ~ 500nm.
The present invention is the corresponding preparation method providing a kind of described complex carrier also, comprises the steps: steps A, preparation acrylamide, methylene diacrylamide and nanometer Fe
3o
4mixed solution; Step B, adds tensio-active agent, cosurfactant and oil phase and obtains reverse micro emulsion in above-mentioned mixed solution; Step C, adds the catalyzer for generating polyacrylamide gel and accelerator in this reverse micro emulsion, the complex carrier of polyreaction product nucleus shell structure; Step D, has the nano combined carrier of superparamagnetism after solid-liquid separation and purifying (selectively dry) described in obtaining.
In the catalyzer used in the present invention and accelerator and prior art, conventional being polymerized by raw material propylene acid amides and methylene diacrylamide generates polyacrylamide gel catalyzer used and accelerator.
In the present invention, by nanometer Fe
3o
4on core basis, use reverse microemulsion process to prepare polyacrylamide gel shell thereon, thus obtain the nano combined carrier described in the present invention.
In a kind of embodiment, described tensio-active agent be cetyl trimethylammonium bromide, bromination dodecyl trimethyl ammonium, Sodium dodecylbenzene sulfonate, styrylphenol formaldehyde resin, polyoxyethylenesorbitan sorbitan monooleate, polyoxyethylene polyoxypropylene block type polyethers, polyoxyethylene alkylphenol ether, polyoxyethylene octylphenol ether, polyoxyethylene nonylphenol ether, aliphatic alcohol polyethenoxy, sulfonic acid esters, amides, silicone based in one or more, preferred cetyl trimethylammonium bromide; Described cosurfactant is alcoholic solvent, one or more in preferred Virahol, propyl alcohol and ethanol; Described oil phase is varsol, is preferably one or more in hexanaphthene, normal heptane and tetracol phenixin.
The nano composite material that the method for the invention prepares has superparamagnetism, and under the driving of externally-applied magnetic field, the enzyme that can carry institute's load carries out directed driven.Compared with traditional fixed enzyme vector, the present invention can realize the displacement of immobilized enzyme, shortens recovery time, greatly improves organic efficiency.Nano composite material particle diameter of the present invention is little, monodispersity good, pore size is controlled, specific surface area is large, chemical stability is good, carrying capacity is strong, good biocompatibility, magnetic response are strong, rate and the enzyme rate of recovery of living is fixed high, catalytic activity and substrate tolerance is strong, cycle index is many, recovery is convenient by the immobilized enzyme of this material prepared by carrier.Preparation method's reaction conditions of the present invention is gentle, and preparation technology is simple, with low cost.
A kind of nano composite material with superparamagnetism of the present invention can be used for preparing immobilized enzyme, but is not limited to immobilized enzyme, also can as the carrier of immobilized cell, sustained and controlled release medicament and targeted drug.
Accompanying drawing explanation
Fig. 1 ~ 8 are respectively the nanometer Fe that embodiment 1 ~ 8 prepares
3o
4the particle size distribution figure of/PAAG matrix material;
Fig. 9 is the nanometer Fe that embodiment 6 prepares
3o
4the scanning electron microscope (SEM) photograph of/PAAG matrix material.
Embodiment
Below in conjunction with specific embodiments and the drawings, the invention will be further described, but the present invention is not restricted to following examples.
The laser particle size analyzer used in embodiment for ZS 90 laser particle size analyzer, scanning electron microscope be S-800 scanning electronic microscope.
Raw materials used source in embodiment: nanometer Fe
3o
4, polyacrylamide, methylene diacrylamide and cetyl trimethylammonium bromide (CTAB) purchase in Sigma company, all the other all reagent and material are all purchased in Aladdin Reagent Company.
Embodiment 1
10mL deionized water, 0.5g nanometer Fe is added in the triangular flask of cleaning
3o
4, 1.0
gpolyacrylamide, 0.05g methylene diacrylamide, completely after mixing, add 10gCTAB, 20mL Virahol and 30mL hexanaphthene respectively, is stirred well to dissolve completely to form stable w/o type reverse micro emulsion; Then 0.010g ammonium persulphate (APS) and 10 μ L Tetramethyl Ethylene Diamines (TEMED) are dissolved in 1.0mL water, after dissolving completely, use constant flow pump, with the flow velocity of 0.2mL/min, this mixed solution is joined in reverse micro emulsion.All add rear continuation stirring reaction 1h to terminate to gel reaction, homodisperse nanometer Fe can be obtained
3o
4/ PAAG suspension.
By nanometer Fe
3o
4/ PAAG suspension is placed in supercentrifuge, with the centrifugal 5min of the speed of 10000r/min, collects centrifugal sediment; Then add water respectively and Virahol carries out cleaning centrifugal sediment, again with the centrifugal 5min of the speed of 10000r/min after cleaning, repeatable operation three times is to nanometer Fe
3o
4the impurity of/PAAG composite material surface and internal residual is washed completely.Then centrifugal sediment is placed in vacuum freeze drier lyophilize, pure nanometer Fe can be obtained
3o
4/ PAAG matrix material.
Scanning electron microscope and laser particle size analyzer measure and show that the nano combined carrier of gained is uniform spherome, and its median size is 32nm.
Embodiment 2
10mL deionized water, 0.5g nanometer Fe is added in the triangular flask of cleaning
3o
4, 3.0g polyacrylamide, 0.05g methylene diacrylamide, completely after mixing, add 10gCTAB, 20mL Virahol and 30mL hexanaphthene respectively, be stirred well to dissolve completely and form stable w/o type reverse micro emulsion; Then 0.030g ammonium persulphate (APS) and 30 μ L Tetramethyl Ethylene Diamines (TEMED) are dissolved in 1.0mL water, after dissolving completely, use constant flow pump, with the flow velocity of 0.2mL/min, this mixed solution is joined in reverse micro emulsion.All add rear continuation stirring reaction 1h to terminate to gel reaction, homodisperse nanometer Fe can be obtained
3o
4/ PAAG suspension.
By nanometer Fe
3o
4/ PAAG suspension is placed in supercentrifuge, with the centrifugal 5min of the speed of 10000r/min, collects centrifugal sediment; Then add water respectively and Virahol carries out cleaning centrifugal sediment, again with the centrifugal 5min of the speed of 10000r/min after cleaning, repeatable operation three times is to nanometer Fe
3o
4the impurity of/PAAG composite material surface and internal residual is washed completely.Then centrifugal sediment is placed in vacuum freeze drier lyophilize, pure nanometer Fe can be obtained
3o
4/ PAAG matrix material.
Scanning electron microscope and laser particle size analyzer measure and show that the nano combined carrier of gained is uniform spherome, and its median size is 128nm.
Embodiment 3
10mL deionized water, 0.5g nanometer Fe is added in the triangular flask of cleaning
3o
4, 5.0g polyacrylamide, 0.05g methylene diacrylamide, completely after mixing, add 10gCTAB, 20mL Virahol and 30mL hexanaphthene respectively, be stirred well to dissolve completely and form stable w/o type reverse micro emulsion; Then 0.050g ammonium persulphate (APS) and 50 μ L Tetramethyl Ethylene Diamines (TEMED) are dissolved in 1.0mL water, after dissolving completely, use constant flow pump, with the flow velocity of 0.2mL/min, this mixed solution is joined in reverse micro emulsion.All add rear continuation stirring reaction 1h to terminate to gel reaction, homodisperse nanometer Fe can be obtained
3o
4/ PAAG suspension.
By nanometer Fe
3o
4/ PAAG suspension is placed in supercentrifuge, with the centrifugal 5min of the speed of 10000r/min, collects centrifugal sediment; Then add water respectively and Virahol carries out cleaning centrifugal sediment, again with the centrifugal 5min of the speed of 10000r/min after cleaning, repeatable operation three times is to nanometer Fe
3o
4the impurity of/PAAG composite material surface and internal residual is washed completely.Then centrifugal sediment is placed in vacuum freeze drier lyophilize, pure nanometer Fe can be obtained
3o
4/ PAAG matrix material.
Scanning electron microscope and laser particle size analyzer measure and show that the nano combined carrier of gained is uniform spherome, and its median size is 399nm.
Embodiment 4
10mL deionized water, 0.5g nanometer Fe is added in the triangular flask of cleaning
3o
4, 10.0g polyacrylamide, 0.05g methylene diacrylamide, completely after mixing, add 10gCTAB, 20mL Virahol and 30mL hexanaphthene respectively, be stirred well to dissolve completely and form stable w/o type reverse micro emulsion; Then 0.10g ammonium persulphate (APS) and 100 μ L Tetramethyl Ethylene Diamines (TEMED) are dissolved in 1.0mL water, after dissolving completely, use constant flow pump, with the flow velocity of 0.2mL/min, this mixed solution is joined in reverse micro emulsion.All add rear continuation stirring reaction 1h to terminate to gel reaction, homodisperse nanometer Fe can be obtained
3o
4/ PAAG suspension.
By nanometer Fe
3o
4/ PAAG suspension is placed in supercentrifuge, with the centrifugal 5min of the speed of 10000r/min, collects centrifugal sediment; Then add water respectively and Virahol carries out cleaning centrifugal sediment, again with the centrifugal 5min of the speed of 10000r/min after cleaning, repeatable operation three times is to nanometer Fe
3o
4the impurity of/PAAG composite material surface and internal residual is washed completely.Then centrifugal sediment is placed in vacuum freeze drier lyophilize, pure nanometer Fe can be obtained
3o
4/ PAAG matrix material.
Scanning electron microscope and laser particle size analyzer measure and show that the nano combined carrier of gained is uniform spherome, and its median size is 647nm.
Embodiment 5
10mL deionized water, 0.5g nanometer Fe is added in the triangular flask of cleaning
3o
4, 1.0g polyacrylamide, 0.2g methylene diacrylamide, completely after mixing, add 10gCTAB, 20mL Virahol and 30mL hexanaphthene respectively, be stirred well to dissolve completely and form stable w/o type reverse micro emulsion; Then 0.100g ammonium persulphate (APS) and 100 μ L Tetramethyl Ethylene Diamines (TEMED) are dissolved in 1.0mL water, after dissolving completely, use constant flow pump, with the flow velocity of 0.2mL/min, this mixed solution is joined in reverse micro emulsion.All add rear continuation stirring reaction 1h to terminate to gel reaction, homodisperse nanometer Fe can be obtained
3o
4/ PAAG suspension.
By nanometer Fe
3o
4/ PAAG suspension is placed in supercentrifuge, with the centrifugal 5min of the speed of 10000r/min, collects centrifugal sediment; Then add water respectively and Virahol carries out cleaning centrifugal sediment, again with the centrifugal 5min of the speed of 10000r/min after cleaning, repeatable operation three times is to nanometer Fe
3o
4the impurity of/PAAG composite material surface and internal residual is washed completely.Then centrifugal sediment is placed in vacuum freeze drier lyophilize, pure nanometer Fe can be obtained
3o
4/ PAAG matrix material.
Scanning electron microscope and laser particle size analyzer measure and show that the nano combined carrier of gained is uniform spherome, and its median size is 12nm.
Embodiment 6
10mL deionized water, 0.5g nanometer Fe is added in the triangular flask of cleaning
3o
4, 3.0g polyacrylamide, 0.2g methylene diacrylamide, completely after mixing, add 10gCTAB, 20mL Virahol and 30mL hexanaphthene respectively, be stirred well to dissolve completely and form stable w/o type reverse micro emulsion; Then 0.030g ammonium persulphate (APS) and 30 μ L Tetramethyl Ethylene Diamines (TEMED) are dissolved in 1.0mL water, after dissolving completely, use constant flow pump, with the flow velocity of 0.2mL/min, this mixed solution is joined in reverse micro emulsion.All add rear continuation stirring reaction 1h to terminate to gel reaction, homodisperse nanometer Fe can be obtained
3o
4/ PAAG suspension.
By nanometer Fe
3o
4/ PAAG suspension is placed in supercentrifuge, with the centrifugal 5min of the speed of 10000r/min, collects centrifugal sediment; Then add water respectively and Virahol carries out cleaning centrifugal sediment, again with the centrifugal 5min of the speed of 10000r/min after cleaning, repeatable operation three times is to nanometer Fe
3o
4the impurity of/PAAG composite material surface and internal residual is washed completely.Then centrifugal sediment is placed in vacuum freeze drier lyophilize, pure nanometer Fe can be obtained
3o
4/ PAAG matrix material.
Scanning electron microscope (Fig. 9) and laser particle size analyzer measure and show that the nano combined carrier of gained is uniform spherome, and its median size is 72nm.
Embodiment 7
10mL deionized water, 0.5g nanometer Fe is added in the triangular flask of cleaning
3o
4, 5.0g polyacrylamide, 0.2g methylene diacrylamide, completely after mixing, add 10gCTAB, 20mL Virahol and 30mL hexanaphthene respectively, be stirred well to dissolve completely and form stable w/o type reverse micro emulsion; Then 0.050g ammonium persulphate (APS) and 50 μ L Tetramethyl Ethylene Diamines (TEMED) are dissolved in 1.0mL water, after dissolving completely, use constant flow pump, with the flow velocity of 0.2mL/min, this mixed solution is joined in reverse micro emulsion.All add rear continuation stirring reaction 1h to terminate to gel reaction, homodisperse nanometer Fe can be obtained
3o
4/ PAAG suspension.
By nanometer Fe
3o
4/ PAAG suspension is placed in supercentrifuge, with the centrifugal 5min of the speed of 10000r/min, collects centrifugal sediment; Then add water respectively and Virahol carries out cleaning centrifugal sediment, again with the centrifugal 5min of the speed of 10000r/min after cleaning, repeatable operation three times is to nanometer Fe
3o
4the impurity of/PAAG composite material surface and internal residual is washed completely.Then centrifugal sediment is placed in vacuum freeze drier lyophilize, pure nanometer Fe can be obtained
3o
4/ PAAG matrix material.
Scanning electron microscope and laser particle size analyzer measure and show that the nano combined carrier of gained is uniform spherome, and its median size is 354nm.
Embodiment 8
10mL deionized water, 0.5g nanometer Fe is added in the triangular flask of cleaning
3o
4, 10.0g polyacrylamide, 0.2g methylene diacrylamide, completely after mixing, add 10gCTAB, 20mL Virahol and 30mL hexanaphthene respectively, be stirred well to dissolve completely and form stable w/o type reverse micro emulsion; Then 0.10g ammonium persulphate (APS) and 100 μ L Tetramethyl Ethylene Diamines (TEMED) are dissolved in 1.0mL water, after dissolving completely, use constant flow pump, with the flow velocity of 0.2mL/min, this mixed solution is joined in reverse micro emulsion.All add rear continuation stirring reaction 1h to terminate to gel reaction, homodisperse nanometer Fe can be obtained
3o
4/ PAAG suspension.
By nanometer Fe
3o
4/ PAAG suspension is placed in supercentrifuge, with the centrifugal 5min of the speed of 10000r/min, collects centrifugal sediment; Then add water respectively and Virahol carries out cleaning centrifugal sediment, again with the centrifugal 5min of the speed of 10000r/min after cleaning, repeatable operation three times is to nanometer Fe
3o
4the impurity of/PAAG composite material surface and internal residual is washed completely.Then centrifugal sediment is placed in vacuum freeze drier lyophilize, pure nanometer Fe can be obtained
3o
4/ PAAG matrix material.
Scanning electron microscope and laser particle size analyzer measure and show that the nano combined carrier of gained is uniform spherome, and its median size is 561nm.
Claims (6)
1. have a nano combined carrier for superparamagnetism, it is characterized in that, described carrier is with nanometer Fe
3o
4be shell for core, with polyacrylamide gel.
2. complex carrier according to claim 1, is characterized in that, the described nanometer Fe as core
3o
4particle diameter be 5 ~ 1000nm, be preferably 10 ~ 200nm, be more preferably 20 ~ 50nm; The described polyacrylamide gel thickness as shell is 5 ~ 1000nm, is preferably 10 ~ 800nm, is more preferably 30 ~ 500nm.
3. complex carrier according to claim 1, is characterized in that, the particle diameter of described carrier is 15 ~ 1000nm, preferably 25 ~ 800nm, more preferably 50 ~ 500nm.
4. the application of the complex carrier in claims 1 to 3 described in any one in fixed enzyme vector, fixed cell carrier, sustained and controlled release medicament carrier or target medicine carrier.
5., as a preparation method for complex carrier as described in any one in claims 1 to 3, comprise the steps:
Steps A, preparation acrylamide, methylene diacrylamide and nanometer Fe
3o
4mixed solution;
Step B, adds tensio-active agent, cosurfactant and oil phase and obtains reverse micro emulsion in above-mentioned mixed solution;
Step C, adds the catalyzer for generating polyacrylamide gel and accelerator in this reverse micro emulsion, the complex carrier of polyreaction product nucleus shell structure;
Step D, has the nano combined carrier of superparamagnetism after solid-liquid separation and purifying described in obtaining.
6. method according to claim 5, it is characterized in that, described tensio-active agent be cetyl trimethylammonium bromide, bromination dodecyl trimethyl ammonium, Sodium dodecylbenzene sulfonate, styrylphenol formaldehyde resin, polyoxyethylenesorbitan sorbitan monooleate, polyoxyethylene polyoxypropylene block type polyethers, polyoxyethylene alkylphenol ether, polyoxyethylene octylphenol ether, polyoxyethylene nonylphenol ether, aliphatic alcohol polyethenoxy, sulfonic acid esters, amides, silicone based in one or more, preferred cetyl trimethylammonium bromide; Described cosurfactant is alcoholic solvent, one or more in preferred Virahol, propyl alcohol and ethanol; Described oil phase is varsol, is preferably one or more in hexanaphthene, normal heptane and tetracol phenixin.
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