CN117700762A - Core-shell type magnetic composite microsphere and preparation method thereof - Google Patents
Core-shell type magnetic composite microsphere and preparation method thereof Download PDFInfo
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- CN117700762A CN117700762A CN202311518393.0A CN202311518393A CN117700762A CN 117700762 A CN117700762 A CN 117700762A CN 202311518393 A CN202311518393 A CN 202311518393A CN 117700762 A CN117700762 A CN 117700762A
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- 239000004005 microsphere Substances 0.000 title claims abstract description 141
- 239000002131 composite material Substances 0.000 title claims abstract description 63
- 239000011258 core-shell material Substances 0.000 title claims abstract description 53
- 238000002360 preparation method Methods 0.000 title claims abstract description 30
- 239000004793 Polystyrene Substances 0.000 claims abstract description 40
- 229920002223 polystyrene Polymers 0.000 claims abstract description 40
- 125000000524 functional group Chemical group 0.000 claims abstract description 21
- 239000000178 monomer Substances 0.000 claims abstract description 21
- PPBRXRYQALVLMV-UHFFFAOYSA-N Styrene Chemical compound C=CC1=CC=CC=C1 PPBRXRYQALVLMV-UHFFFAOYSA-N 0.000 claims abstract description 15
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims abstract description 15
- 238000000034 method Methods 0.000 claims abstract description 15
- 239000004372 Polyvinyl alcohol Substances 0.000 claims abstract description 13
- 229920002451 polyvinyl alcohol Polymers 0.000 claims abstract description 13
- 239000006087 Silane Coupling Agent Substances 0.000 claims abstract description 11
- 238000006482 condensation reaction Methods 0.000 claims abstract description 11
- 239000003999 initiator Substances 0.000 claims abstract description 11
- 238000011065 in-situ storage Methods 0.000 claims abstract description 9
- 239000003381 stabilizer Substances 0.000 claims abstract description 8
- 238000001308 synthesis method Methods 0.000 claims abstract description 7
- 230000007062 hydrolysis Effects 0.000 claims abstract description 6
- 238000006460 hydrolysis reaction Methods 0.000 claims abstract description 6
- 230000009471 action Effects 0.000 claims abstract description 3
- 230000000379 polymerizing effect Effects 0.000 claims abstract description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 87
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 24
- 238000003756 stirring Methods 0.000 claims description 24
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 20
- 238000006243 chemical reaction Methods 0.000 claims description 19
- OZAIFHULBGXAKX-UHFFFAOYSA-N 2-(2-cyanopropan-2-yldiazenyl)-2-methylpropanenitrile Chemical compound N#CC(C)(C)N=NC(C)(C)C#N OZAIFHULBGXAKX-UHFFFAOYSA-N 0.000 claims description 17
- 238000010438 heat treatment Methods 0.000 claims description 14
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 14
- 238000006116 polymerization reaction Methods 0.000 claims description 13
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 10
- 239000002612 dispersion medium Substances 0.000 claims description 8
- 229920000036 polyvinylpyrrolidone Polymers 0.000 claims description 8
- 239000001267 polyvinylpyrrolidone Substances 0.000 claims description 8
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 claims description 8
- 230000003301 hydrolyzing effect Effects 0.000 claims description 7
- 239000007800 oxidant agent Substances 0.000 claims description 7
- 239000007864 aqueous solution Substances 0.000 claims description 6
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims description 6
- 230000035484 reaction time Effects 0.000 claims description 6
- XDLMVUHYZWKMMD-UHFFFAOYSA-N 3-trimethoxysilylpropyl 2-methylprop-2-enoate Chemical compound CO[Si](OC)(OC)CCCOC(=O)C(C)=C XDLMVUHYZWKMMD-UHFFFAOYSA-N 0.000 claims description 5
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 5
- 230000001590 oxidative effect Effects 0.000 claims description 5
- 239000000243 solution Substances 0.000 claims description 5
- 229960002089 ferrous chloride Drugs 0.000 claims description 4
- 238000003018 immunoassay Methods 0.000 claims description 4
- NMCUIPGRVMDVDB-UHFFFAOYSA-L iron dichloride Chemical compound Cl[Fe]Cl NMCUIPGRVMDVDB-UHFFFAOYSA-L 0.000 claims description 4
- WYTZZXDRDKSJID-UHFFFAOYSA-N (3-aminopropyl)triethoxysilane Chemical compound CCO[Si](OCC)(OCC)CCCN WYTZZXDRDKSJID-UHFFFAOYSA-N 0.000 claims description 3
- 239000005708 Sodium hypochlorite Substances 0.000 claims description 3
- 239000011790 ferrous sulphate Substances 0.000 claims description 3
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 3
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 3
- 235000010333 potassium nitrate Nutrition 0.000 claims description 3
- 239000004323 potassium nitrate Substances 0.000 claims description 3
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims description 3
- 125000000542 sulfonic acid group Chemical group 0.000 claims description 3
- BPSIOYPQMFLKFR-UHFFFAOYSA-N trimethoxy-[3-(oxiran-2-ylmethoxy)propyl]silane Chemical compound CO[Si](OC)(OC)CCCOCC1CO1 BPSIOYPQMFLKFR-UHFFFAOYSA-N 0.000 claims description 3
- 238000009007 Diagnostic Kit Methods 0.000 claims description 2
- 239000011203 carbon fibre reinforced carbon Substances 0.000 claims description 2
- 238000009833 condensation Methods 0.000 claims description 2
- 230000005494 condensation Effects 0.000 claims description 2
- IMBKASBLAKCLEM-UHFFFAOYSA-L ferrous ammonium sulfate (anhydrous) Chemical compound [NH4+].[NH4+].[Fe+2].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O IMBKASBLAKCLEM-UHFFFAOYSA-L 0.000 claims description 2
- USHAGKDGDHPEEY-UHFFFAOYSA-L potassium persulfate Chemical compound [K+].[K+].[O-]S(=O)(=O)OOS([O-])(=O)=O USHAGKDGDHPEEY-UHFFFAOYSA-L 0.000 claims description 2
- 238000010526 radical polymerization reaction Methods 0.000 claims description 2
- 239000002245 particle Substances 0.000 abstract description 17
- 230000008569 process Effects 0.000 abstract description 5
- 238000001514 detection method Methods 0.000 abstract description 3
- 239000000090 biomarker Substances 0.000 abstract 1
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- 238000006557 surface reaction Methods 0.000 abstract 1
- 235000019441 ethanol Nutrition 0.000 description 26
- 239000004593 Epoxy Substances 0.000 description 15
- 238000005406 washing Methods 0.000 description 13
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 12
- 230000036571 hydration Effects 0.000 description 7
- 238000006703 hydration reaction Methods 0.000 description 7
- 239000011324 bead Substances 0.000 description 6
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- 239000012046 mixed solvent Substances 0.000 description 6
- 230000004048 modification Effects 0.000 description 6
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- 229910052757 nitrogen Inorganic materials 0.000 description 6
- 150000001336 alkenes Chemical class 0.000 description 5
- 238000001035 drying Methods 0.000 description 5
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- JRZJOMJEPLMPRA-UHFFFAOYSA-N olefin Natural products CCCCCCCC=C JRZJOMJEPLMPRA-UHFFFAOYSA-N 0.000 description 5
- 238000012512 characterization method Methods 0.000 description 4
- 238000009826 distribution Methods 0.000 description 4
- 239000011259 mixed solution Substances 0.000 description 4
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- 238000007306 functionalization reaction Methods 0.000 description 3
- 238000004020 luminiscence type Methods 0.000 description 3
- 239000006249 magnetic particle Substances 0.000 description 3
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- CERQOIWHTDAKMF-UHFFFAOYSA-N Methacrylic acid Chemical compound CC(=C)C(O)=O CERQOIWHTDAKMF-UHFFFAOYSA-N 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000006978 adaptation Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
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- 238000001000 micrograph Methods 0.000 description 2
- 239000002105 nanoparticle Substances 0.000 description 2
- 230000020477 pH reduction Effects 0.000 description 2
- 229920001467 poly(styrenesulfonates) Polymers 0.000 description 2
- 229960002796 polystyrene sulfonate Drugs 0.000 description 2
- 239000011970 polystyrene sulfonate Substances 0.000 description 2
- MNCGMVDMOKPCSQ-UHFFFAOYSA-M sodium;2-phenylethenesulfonate Chemical compound [Na+].[O-]S(=O)(=O)C=CC1=CC=CC=C1 MNCGMVDMOKPCSQ-UHFFFAOYSA-M 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000009210 therapy by ultrasound Methods 0.000 description 2
- DZSVIVLGBJKQAP-UHFFFAOYSA-N 1-(2-methyl-5-propan-2-ylcyclohex-2-en-1-yl)propan-1-one Chemical compound CCC(=O)C1CC(C(C)C)CC=C1C DZSVIVLGBJKQAP-UHFFFAOYSA-N 0.000 description 1
- MTEZSDOQASFMDI-UHFFFAOYSA-N 1-trimethoxysilylpropan-1-ol Chemical compound CCC(O)[Si](OC)(OC)OC MTEZSDOQASFMDI-UHFFFAOYSA-N 0.000 description 1
- SMZOUWXMTYCWNB-UHFFFAOYSA-N 2-(2-methoxy-5-methylphenyl)ethanamine Chemical compound COC1=CC=C(C)C=C1CCN SMZOUWXMTYCWNB-UHFFFAOYSA-N 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-N 2-Propenoic acid Natural products OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 1
- LBSXSAXOLABXMF-UHFFFAOYSA-N 4-Vinylaniline Chemical compound NC1=CC=C(C=C)C=C1 LBSXSAXOLABXMF-UHFFFAOYSA-N 0.000 description 1
- BTBUEUYNUDRHOZ-UHFFFAOYSA-N Borate Chemical compound [O-]B([O-])[O-] BTBUEUYNUDRHOZ-UHFFFAOYSA-N 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000010835 comparative analysis Methods 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
- 238000007334 copolymerization reaction Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000011261 inert gas Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000006148 magnetic separator Substances 0.000 description 1
- FPYJFEHAWHCUMM-UHFFFAOYSA-N maleic anhydride Chemical compound O=C1OC(=O)C=C1 FPYJFEHAWHCUMM-UHFFFAOYSA-N 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
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- 239000002086 nanomaterial Substances 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 235000012239 silicon dioxide Nutrition 0.000 description 1
- SONHXMAHPHADTF-UHFFFAOYSA-M sodium;2-methylprop-2-enoate Chemical compound [Na+].CC(=C)C([O-])=O SONHXMAHPHADTF-UHFFFAOYSA-M 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
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- Manufacturing Of Micro-Capsules (AREA)
Abstract
The invention discloses a core-shell type magnetic composite microsphere and a preparation method thereof. The method comprises the following steps: polymerizing styrene and a stabilizer for the first time under the action of an initiator, and then adding a functional group polymerizable monomer for the second time to obtain polystyrene functional microspheres; preparing magnetic microspheres by using ferrous salt and polystyrene functional microspheres through an in-situ synthesis method; the core-shell type magnetic composite microsphere is prepared by the hydrolysis condensation reaction of polyvinyl alcohol, a silane coupling agent and the magnetic microsphere. The preparation method provided by the invention has simple process and universality, and the prepared core-shell type magnetic composite microsphere has a compact magnetic layer with high crystallinity, controllable particle size, easy surface functionalization and good biocompatibility, can be used as a biological sensing material and is applied to biomarker detection.
Description
Technical Field
The invention relates to the technical field of magnetic micro-nano materials, in particular to a core-shell type magnetic composite microsphere and a preparation method thereof.
Background
The stability of the composite interface of the polystyrene microsphere and the magnetic layer is the key for preparing the core-shell type magnetic composite microsphere. The method for improving the interaction force of the composite interface adopts a mode of modifying the surface of the polystyrene microsphere and constructing functional magnetic particles. Wherein, the surface modification of polystyrene microsphere usually uses strong acid acidification treatment, the modification process is complex, the process is difficult to control, and the morphology and the surface structure of the microsphere can be damaged. In addition, the mode of preparing functional magnetic particles in advance and then carrying out interface compounding is needed to be carried out in multiple steps, the post-treatment is complex, and the obtained microsphere magnetic layer is not compact and is easy to generate magnetic leakage.
Disclosure of Invention
The invention provides a core-shell type magnetic composite microsphere and a preparation method thereof. The method adopts a mode of copolymerizing functional group monomers to realize the functionalization of the polystyrene microsphere, and then constructs a compact magnetic layer with high crystallinity by an in-situ synthesis mode, thereby avoiding the magnetic leakage phenomenon; finally, the magnetic composite microsphere is constructed through hydrolysis condensation reaction.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
in one aspect, the invention provides a method for preparing a core-shell type magnetic composite microsphere, comprising the following steps:
s1: polymerizing styrene and a stabilizer for the first time under the action of an initiator, and then adding a functional group polymerizable monomer for the second time to obtain polystyrene functional microspheres;
s2: preparing magnetic microspheres by using a ferrous salt and the polystyrene functional microspheres obtained in the step S1 through an in-situ synthesis method;
s3: and (2) preparing the core-shell type magnetic composite microsphere by a hydrolysis condensation reaction of the polyvinyl alcohol, the silane coupling agent and the magnetic microsphere obtained in the step (S2).
As a preferred embodiment, in step S1, the functional group-polymerizable monomer is selected from any one of polymerizable monomers containing a carboxyl functional group, an amino functional group, and a sulfonic acid functional group;
specifically, the polymerizable monomer having a carboxyl functional group includes acrylic acid, methacrylic acid, maleic anhydride, and the like; the polymerizable monomer having an amino functional group includes 4-aminostyrene, acrylamide, N-methylenebisacrylamide, and the like; examples of the polymerizable monomer having a sulfonic acid functional group include sodium styrene sulfonate, sodium methacrylate sulfonate, and 2-acrylamide-2-methylpropanesulfonic acid; the above list can be used alone or in combination with the functional groups of the same kind;
preferably, the polymerizable monomer is a monomer containing a carbon-carbon double bond and capable of undergoing free radical polymerization;
preferably, the stabilizer is polyvinylpyrrolidone;
preferably, the initiator is any one of Azobisisobutyronitrile (AIBN) or potassium persulfate (KPS);
preferably, the primary and secondary polymerizations are carried out in a dispersion medium; the dispersion medium is preferably ethanol;
preferably, the amount of styrene is 8 to 16g, the amount of stabilizer is 0.3 to 2g, the amount of initiator is 0.08 to 0.5g, and the amount of functional group polymerizable monomer is 0.8 to 3g, based on 100g of the dispersion medium.
In a preferred embodiment, in the step S1, the reaction temperature of the primary polymerization is 60 to 80 ℃, and the reaction time of the primary polymerization is 5 to 8 hours;
preferably, the reaction temperature of the secondary polymerization is 60-80 ℃, and the reaction time of the secondary polymerization is 3-5 h;
preferably, the primary and secondary polymerizations are carried out under stirring conditions;
preferably, the stirring speed is 150-250 rpm.
In certain specific embodiments, step S1 further comprises a post-treatment operation comprising washing and drying.
In certain specific embodiments, the specific operation of step S1 includes the steps of:
dispersing a stabilizer in a dispersion medium under inert atmosphere, and heating to 60-80 ℃; under the stirring condition of 150-250 rpm, adding a mixture of an initiator and styrene, and then carrying out polymerization reaction for 5-8 h; adding the mixed solution of the functional group polymerizable monomer and the dispersion medium to continue to polymerize for 3-5 h; washing with ethanol and drying to obtain the polystyrene functional microsphere.
In a preferred embodiment, in step S2, the in situ synthesis method comprises the steps of:
in an inert atmosphere, under the stirring condition, heating the polystyrene functional microspheres prepared in the step S1 with sodium hydroxide, an oxidant and ferrous salt in an aqueous solution for reaction;
preferably, the oxidizing agent is selected from any one of hydrogen peroxide, potassium nitrate and sodium hypochlorite;
preferably, the ferrous salt is selected from at least one of ferrous sulfate, ferrous ammonium sulfate and ferrous chloride;
preferably, the mass ratio of the polystyrene functional microsphere to sodium hydroxide to ferrous salt is 1:2 to 6:3.5 to 7.5;
preferably, the mass of the oxidant is 2-10 times of that of ferrous salt;
preferably, the temperature of the heating reaction is 60-100 ℃, and the time of the heating reaction is 2-24 hours;
preferably, the stirring speed is 250-350 rpm;
in certain specific embodiments, step S2 further comprises a post-treatment comprising washing.
In certain embodiments, step S2 specifically comprises the steps of: dispersing the polystyrene functional microspheres obtained in the step S1 in water, and stirring for 0.5-1.5 h; introducing inert gas, sequentially adding sodium hydroxide and an oxidant, and stirring for 0.5-1 h; heating to 60-100 ℃, and rapidly adding ferrous salt aqueous solution for reacting for 2-24 h; washing with pure water and ethanol to obtain the magnetic microsphere;
in a preferred embodiment, in step S3, the hydrolytic condensation reaction is specifically: carrying out hydrolytic condensation reaction on the magnetic microspheres obtained in the step S2, polyvinyl alcohol and a silane coupling agent in an ethanol solution;
preferably, the molecular weight of the polyvinyl alcohol is 5000-35000;
preferably, the silane coupling agent is selected from at least one of 3-aminopropyl triethoxysilane (KH-550), gamma-glycidoxypropyl trimethoxysilane (KH-560) and gamma-methacryloxypropyl trimethoxysilane (KH-570);
preferably, the mass ratio of the magnetic microsphere to the polyvinyl alcohol to the silane coupling agent is 1:0.5 to 2.5:1 to 10;
preferably, in the ethanol solution, the volume ratio of ethanol to water is 1: 5-20;
preferably, the reaction temperature of the hydrolytic condensation is 50-90 ℃ and the reaction time is 12-24 hours;
in certain specific embodiments, step S3 further comprises a post-treatment operation, said post-treatment comprising washing.
In certain embodiments, step S3 specifically comprises the steps of: dispersing the magnetic microspheres in an ethanol solution under the stirring condition; adding polyvinyl alcohol and stirring for 0.5-3 h by ultrasonic; adding a silane coupling agent, stirring for 0.5-1 h by ultrasonic, heating to 50-90 ℃ and continuing to react for 12-24 h; washing with pure water and ethanol to obtain the core-shell type magnetic composite microsphere.
In yet another aspect, the invention provides the core-shell type magnetic composite microsphere obtained by the preparation method.
In the technical scheme of the invention, amino, epoxy, olefin and other corresponding functional groups are modified on the surface of the core-shell magnetic composite microsphere.
In still another aspect, the invention provides an application of the core-shell type magnetic composite microsphere in preparing a chemiluminescent immunoassay diagnostic kit.
In the technical scheme of the invention, the core-shell type magnetic composite microsphere can be used for constructing immunomagnetic beads and is further applied to chemiluminescence immunoassay diagnosis.
The technical scheme has the following advantages or beneficial effects:
the invention provides a core-shell type magnetic composite microsphere and a preparation method thereof, wherein the method adopts a mode of copolymerizing functional group monomers to realize the functionalization of polystyrene microspheres; then a compact magnetic layer with high crystallinity is constructed in an in-situ synthesis mode, so that the phenomenon of magnetic leakage is avoided; finally, the magnetic composite microsphere is constructed through hydrolysis condensation reaction. The magnetic composite microsphere prepared by the invention has controllable particle size and universality: firstly, the density of functional groups on the surface of the polystyrene microsphere can be regulated and controlled by changing the types and the quantity of functional group monomers, so that the preparation of the magnetic microsphere with different magnetic contents is realized; secondly, the preparation of magnetic composite microspheres with different functions can be realized by changing the types and the dosage of the silane coupling agent, and the preparation is oriented to different biosensing applications.
Compared with the prior art, the invention has the following advantages:
according to the preparation method of the core-shell type magnetic composite microsphere, the functionalization of the polystyrene microsphere is realized by the copolymerization of the functional group monomers, the strong acid acidification treatment of the microsphere surface is avoided, the morphology and the surface structure of the microsphere are not damaged, the preparation method has the advantages of simple process and controllable preparation, and the industrial mass production is facilitated.
According to the preparation method of the core-shell type magnetic composite microsphere, disclosed by the invention, the compact magnetic layer can be constructed on the surface of the polystyrene functional microsphere through the in-situ synthesis method, so that the core-shell type magnetic composite microsphere with different functionalities can be prepared by using the compact magnetic layer as an ideal matrix, and the magnetic particle prepared on the basis of the in-situ synthesis method has the advantages of good size uniformity, high crystallinity, good magnetic response performance and the like, and is beneficial to the stable batch preparation of the magnetic composite microsphere.
According to the preparation method of the core-shell type magnetic composite microsphere with controllable particle size, provided by the invention, the multifunctional magnetic composite microsphere with controllable magnetic content and functional group distribution is constructed through hydrolysis condensation reaction, so that magnetic leakage of the magnetic microsphere is avoided, meanwhile, silicon dioxide coating is not needed, the process is simple, the advantages of simplicity in operation, mild conditions and universality in scheme are achieved, and the preparation method can be applied to different biosensing.
Drawings
Fig. 1: scanning electron microscope pictures of polystyrene functional microspheres prepared in the embodiments 1-3 of the invention;
fig. 2: the hydration particle size distribution diagram of the polystyrene functional microspheres prepared in the embodiments 1-3 of the invention;
fig. 3: a transmission electron microscope image of the magnetic microsphere prepared in the embodiment 2 of the invention;
fig. 4: the transmission electron microscope image of the core-shell type magnetic composite microsphere prepared in the embodiment 2 of the invention;
fig. 5: the hydration particle size distribution diagram of the core-shell type magnetic composite microsphere prepared in the embodiment 2 of the invention;
fig. 6: surface charge patterns of core-shell magnetic composite microspheres prepared in examples 1-3 of the invention;
fig. 7: magnetic response performance evaluation diagrams of core-shell magnetic composite microspheres prepared in examples 1-3 of the invention.
Detailed Description
The following examples are only some, but not all, of the examples of the invention. Accordingly, the detailed description of the embodiments of the invention provided below is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be made by a person skilled in the art without making any inventive effort, are intended to fall within the scope of the present invention.
In the present invention, all the equipment, raw materials and the like are commercially available or commonly used in the industry unless otherwise specified. The methods in the following examples are conventional in the art unless otherwise specified.
Example 1: amino core-shell type magnetic composite microsphere
The embodiment provides an amino core-shell type magnetic composite microsphere, which is prepared by the following steps:
s1: preparation of polystyrene carboxyl microsphere
2.5g of polyvinylpyrrolidone (PVP) and 250g of ethanol are added into a three-neck flask, nitrogen is introduced, and the temperature is raised to 75 ℃; after 0.5g of initiator Azobisisobutyronitrile (AIBN) and 25g of styrene monomer were mixed, the mixture was stirred at 200rpm and added to a three-necked flask to react for 6 hours; then adding a mixed solution of 3g of methacrylic acid and 15g of ethanol for continuous reaction for 4 hours; washing with ethanol for 3 times, and drying at 50 ℃ to obtain polystyrene carboxyl microspheres;
s2: preparation of magnetic microspheres
Adding 1g of the polystyrene carboxyl microsphere obtained in the step S1 and 90g of pure water into a three-neck flask, and stirring at 250rpm for 0.5h; introducing nitrogen, sequentially adding 5g of sodium hydroxide and 40mL of 30% hydrogen peroxide solution, stirring for 30min, heating to 80 ℃, rapidly adding 10mL of 0.5g/mL ferrous sulfate aqueous solution, and continuing to react for 6h; washing with pure water and ethanol for 3 times to obtain magnetic microsphere dispersion;
s3: preparing the core-shell type magnetic composite microsphere.
Performing magnetic separation on the magnetic microsphere dispersion liquid to obtain 1g of magnetic microspheres, then re-dispersing the magnetic microspheres in a mixed solvent of 10mL of ethanol and 80mL of pure water, adding the mixed solvent into a three-neck flask, and stirring at 300rpm for 0.5h; and (3) starting water bath ultrasonic treatment, adding 1g of polyvinyl alcohol (with the molecular weight of 9000) into a three-neck flask, carrying out ultrasonic stirring reaction for 1h, then heating to 60 ℃, adding 3g of 3-aminopropyl triethoxysilane (KH-550), carrying out reaction for 12h, and then cleaning for 3 times by using pure water and ethanol respectively to obtain the amino core-shell type magnetic composite microsphere.
Example 2: epoxy core-shell type magnetic composite microsphere
The embodiment provides an epoxy-based core-shell type magnetic composite microsphere, which is prepared by the following steps:
s1: preparation of polystyrene amino microspheres
2.5g of polyvinylpyrrolidone (PVP) and 250g of ethanol are added into a three-neck flask, nitrogen is introduced, and the temperature is raised to 75 ℃; after 0.5g of initiator Azobisisobutyronitrile (AIBN) and 25g of styrene monomer were mixed, the mixture was stirred at 200rpm and added to a three-necked flask to react for 7 hours; then adding a mixed solution of 4g of acrylamide and 14g of ethanol for continuous reaction for 3 hours; washing with ethanol for 3 times, and drying at 50 ℃ to obtain polystyrene amino microspheres;
s2: preparation of magnetic microspheres
2g of polystyrene amino microspheres and 90g of pure water are added into a three-neck flask, and stirred at 250rpm for 0.5h; introducing nitrogen, sequentially adding 6g of sodium hydroxide and 40.4g of potassium nitrate, stirring for 30min, heating to 80 ℃, rapidly adding 10mL of 0.9g/mL ferrous chloride aqueous solution, and continuing to react for 12h; washing with pure water and ethanol for 3 times to obtain magnetic microspheres;
s3: preparing the core-shell type magnetic composite microsphere.
Performing magnetic separation on the magnetic microsphere dispersion liquid to obtain 1g of magnetic microspheres, then re-dispersing the magnetic microspheres in a mixed solvent of 15mL of ethanol and 75mL of pure water, and adding the mixed solvent into a three-neck flask and stirring at 300rpm for 1h; and (3) starting water bath ultrasonic treatment, adding 1.25g of polyvinyl alcohol (with the molecular weight of 15000) into a three-neck flask, carrying out ultrasonic stirring reaction for 1.5h, heating to 80 ℃, adding 8g of gamma-glycidol ether oxypropyl trimethoxysilane (KH-560), carrying out reaction for 16h, and cleaning for 3 times by using pure water and ethanol respectively to obtain the epoxy core-shell type magnetic composite microsphere.
Example 3: olefine base magnetic composite microsphere
The embodiment provides an olefin-based core-shell type magnetic composite microsphere, which is prepared by the following steps:
s1: preparation of polystyrene sulfonate microspheres
2.5g of polyvinylpyrrolidone (PVP) and 250g of ethanol are added into a three-neck flask, nitrogen is introduced, and the temperature is raised to 75 ℃; after 0.5g of initiator Azobisisobutyronitrile (AIBN) and 25g of styrene monomer were mixed, the mixture was stirred at 200rpm and added to a three-necked flask to react for 5 hours; then adding a mixed solution of 5g of sodium styrene sulfonate and 13g of ethanol for continuous reaction for 5 hours; washing with ethanol for 3 times, and drying at 50 ℃ to obtain polystyrene sulfonic microspheres;
s2: preparation of magnetic microspheres
1.5g polystyrene sulfonate microspheres and 90mL pure water are added into a three-neck flask, and stirred at 250rpm for 0.5h; introducing nitrogen, sequentially adding 5g of sodium hydroxide and 29.6g of sodium hypochlorite, stirring for 30min, heating to 90 ℃, rapidly adding 10mL of 0.8g/mL ferrous chloride aqueous solution, and continuing to react for 10h; washing with pure water and ethanol for 3 times to obtain magnetic microspheres;
s3: preparation of olefine base core-shell type magnetic composite microsphere
Performing magnetic separation on the magnetic microsphere dispersion liquid to obtain 1g of magnetic microspheres, then re-dispersing the magnetic microspheres in a mixed solvent of 10mL of ethanol and 80mL of pure water, and adding the mixed solvent into a three-neck flask and stirring at 300rpm for 1h; turning on water bath ultrasonic, adding 2g of polyvinyl alcohol (with molecular weight of 25000) into a three-neck flask, carrying out ultrasonic stirring reaction for 2h, then heating to 70 ℃, adding 6g of gamma-methacryloxypropyl trimethoxy silane (KH-570), carrying out reaction for 24h, and cleaning for 3 times by using pure water and ethanol respectively to obtain the olefin-based core-shell magnetic composite microsphere.
The invention performs morphology characterization on the polystyrene functional microsphere prepared in the step S1 in the examples 1-3 through a scanning electron microscope, and as shown in the figure 1, the polystyrene carboxyl microsphere, the polystyrene amino microsphere and the polystyrene sulfonic microsphere are spherical structures with smooth surfaces, the particle sizes of the polystyrene carboxyl microsphere, the polystyrene amino microsphere and the polystyrene sulfonic microsphere are respectively 1.06+/-0.18 mu m, 1.04+/-0.29 mu m and 1.01+/-0.16 mu m, and the variation coefficient CV value is less than 3%.
The hydration particle size of the polystyrene functional microspheres prepared in the step S1 in the examples 1-3 is characterized by a nanometer particle size potentiometer, and as can be seen from the figure 2, the average hydration particle sizes of the polystyrene carboxyl microspheres, the polystyrene amino microspheres and the polystyrene sulfonic microspheres are respectively 1.48 mu m, 1.45 mu m and 1.43 mu m, and the particle size distribution is uniform.
The magnetic microsphere prepared in the step S2 of the embodiment 2 and the core-shell type magnetic composite microsphere prepared in the step S3 are subjected to morphology characterization by a transmission electron microscope, and the morphology characterization is shown in the figures 3 and 4: (1) The magnetic microsphere has a rough surface and a spherical structure with a particle size of 1.12+/-0.08 mu m; the dense magnetic layer can be seen from the position of the arrow in the figure (fig. 3); (2) The surface of the core-shell type magnetic composite microsphere prepared by the hydrolytic condensation reaction is smooth, and the microsphere has a spherical structure with the particle size of 1.19+/-0.03 mu m, and an obvious modification layer can be seen from the position of an arrow in the figure (figure 4).
The epoxy-based core-shell type magnetic composite microsphere prepared in the embodiment 2 is subjected to hydration particle size characterization by a nano-particle size potentiometer, and as shown in fig. 5, the average hydration particle size of the epoxy-based core-shell type magnetic composite microsphere is 1.57 mu m, and the average hydration particle size is increased by about 120nm relative to the particle size of the magnetic microsphere, so that the surface of the core-shell type magnetic composite microsphere is disclosed to be provided with a modification layer.
According to the invention, the surface charge of the core-shell type magnetic composite microsphere prepared in the examples 1-3 is characterized by a nano-particle size potentiometer, and as can be seen from fig. 6, the surface charges of the prepared amino, epoxy and olefin-based core-shell type magnetic composite microsphere are 17.05+/-0.65 mV, -23.53+/-2.62 mV and-18.23+/-0.89 mV respectively, which indicate that the functional groups are successfully modified on the surface of the core-shell type magnetic composite microsphere.
According to the invention, the magnetic response performance of the core-shell type magnetic composite microsphere is evaluated through the magnetic separation time, as shown in fig. 7, the carboxyl, epoxy and olefin-based core-shell type magnetic composite microsphere prepared in the embodiments 1-3 of the invention is dispersed in absolute ethyl alcohol, and the magnetic separator is used for carrying out magnetic separation on the dispersion liquid, so that three functionalized core-shell type magnetic composite microsphere dispersion liquids can be completely separated within 60 seconds, excellent magnetic response performance is shown, the coating of a magnetic layer is complete, and no magnetic loss occurs.
The invention evaluates the chemiluminescent immunoassay performance of the core-shell type magnetic composite microsphere prepared in the embodiment, and specifically comprises the following steps:
10mg of the epoxy-based core-shell type magnetic composite microsphere prepared in example 2 and commercial epoxy-based magnetic beads (epoxy-based magnetic beads with the particle size of 1 μm of Shanghai Aba Ding Shenghua technology Co.) are respectively taken and dispersed in 1mL of 0.1M borate buffer (pH 8.5); 200 μg of mouse anti-human Thyroid Stimulating Hormone (TSH) monoclonal antibody is added, and after uniform mixing, the mixture is incubated for 24 hours at 37 ℃; add 100. Mu.L of commercial blocking reagent (JSR company Blockmaster CE 210) and continue incubation for 24h; magnetic separation and washing were performed 3 times with 0.1M Tris buffer (pH 7.4) containing 0.1% Tween-20 to obtain magnetic beads coated with mouse anti-human TSH monoclonal antibodies, and further luminescence value detection was performed on a chemiluminescent instrument, and specific detection results are shown in Table 1.
As can be seen from Table 1, the epoxy-based magnetic composite microsphere prepared in example 2 has lower luminescence value of negative sample, higher luminescence value of positive sample, high sensitivity and wide linear range, compared with the commercial epoxy-based magnetic beads.
Table 1 comparative analysis of chemiluminescent detection Performance of epoxy-based core-shell magnetic composite microspheres and commercial epoxy-based magnetic beads prepared in example 2
The foregoing is only a preferred embodiment of the invention, it being noted that: it will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the principles of the present invention, and such modifications and adaptations are intended to be comprehended within the scope of the invention.
Claims (10)
1. The preparation method of the core-shell type magnetic composite microsphere is characterized by comprising the following steps of:
s1: polymerizing styrene and a stabilizer for the first time under the action of an initiator, and then adding a functional group polymerizable monomer for the second time to obtain polystyrene functional microspheres;
s2: preparing magnetic microspheres by using a ferrous salt and the polystyrene functional microspheres obtained in the step S1 through an in-situ synthesis method;
s3: and (2) preparing the core-shell type magnetic composite microsphere by a hydrolysis condensation reaction of the polyvinyl alcohol, the silane coupling agent and the magnetic microsphere obtained in the step (S2).
2. The method according to claim 1, wherein in step S1, the functional group-polymerizable monomer is selected from any one of polymerizable monomers containing a carboxyl functional group, an amino functional group, and a sulfonic acid functional group;
preferably, the polymerizable monomer is a monomer containing a carbon-carbon double bond and capable of undergoing free radical polymerization;
preferably, the stabilizer is polyvinylpyrrolidone;
preferably, the initiator is any one of azodiisobutyronitrile or potassium persulfate;
preferably, the primary and secondary polymerizations are carried out in a dispersion medium; the dispersion medium is preferably ethanol.
3. The preparation method according to claim 2, wherein the amount of styrene is 8 to 16g, the amount of stabilizer is 0.3 to 2g, the amount of initiator is 0.08 to 0.5g, and the amount of functional group polymerizable monomer is 0.8 to 3g based on 100g of the dispersion medium.
4. The preparation method according to claim 1, wherein in the step S1, the reaction temperature of the primary polymerization is 60 to 80 ℃, and the reaction time of the primary polymerization is 5 to 8 hours;
preferably, the reaction temperature of the secondary polymerization is 60-80 ℃, and the reaction time of the secondary polymerization is 3-5 h;
preferably, the primary and secondary polymerizations are carried out under stirring conditions;
preferably, the stirring speed is 150-250 rpm.
5. The method of claim 1, wherein in step S2, the in situ synthesis method comprises the steps of:
and (3) in an inert atmosphere, heating and reacting the polystyrene functional microspheres prepared in the step (S1) with sodium hydroxide, an oxidant and ferrous salt in an aqueous solution under the stirring condition.
6. The method according to claim 5, wherein the oxidizing agent is selected from any one of hydrogen peroxide, potassium nitrate and sodium hypochlorite;
preferably, the ferrous salt is selected from at least one of ferrous sulfate, ferrous ammonium sulfate and ferrous chloride;
preferably, the mass ratio of the polystyrene functional microsphere to sodium hydroxide to ferrous salt is 1:2 to 6:3.5 to 7.5;
preferably, the mass of the oxidant is 2-10 times of the mass of the ferrous salt.
7. The method according to claim 5, wherein the heating reaction is carried out at a temperature of 60 to 100 ℃ for a time of 2 to 24 hours;
preferably, the stirring speed is 250-350 rpm.
8. The preparation method according to claim 1, wherein in step S3, the hydrolytic condensation reaction is specifically: carrying out hydrolytic condensation reaction on the magnetic microspheres obtained in the step S2, polyvinyl alcohol and a silane coupling agent in an ethanol solution;
preferably, the molecular weight of the polyvinyl alcohol is 5000-35000;
preferably, the silane coupling agent is selected from at least one of 3-aminopropyl triethoxysilane, gamma-glycidoxypropyl trimethoxysilane and gamma-methacryloxypropyl trimethoxysilane;
preferably, the mass ratio of the magnetic microsphere to the polyvinyl alcohol to the silane coupling agent is 1:0.5 to 2.5:1 to 10;
preferably, in the ethanol solution, the volume ratio of ethanol to water is 1: 5-20;
preferably, the reaction temperature of the hydrolytic condensation is 50-90 ℃ and the reaction time is 12-24 hours.
9. The core-shell type magnetic composite microsphere obtained by the preparation method of any one of claims 1 to 8.
10. Use of the core-shell type magnetic composite microsphere according to claim 9 for preparing a chemiluminescent immunoassay diagnostic kit.
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