CN114672301B - Preparation method of aggregation-induced emission microsphere with core-shell structure - Google Patents
Preparation method of aggregation-induced emission microsphere with core-shell structure Download PDFInfo
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- 238000004220 aggregation Methods 0.000 title claims abstract description 73
- 239000004005 microsphere Substances 0.000 title claims abstract description 55
- 239000011258 core-shell material Substances 0.000 title claims abstract description 24
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- 229920000642 polymer Polymers 0.000 claims abstract description 20
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- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 6
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- WBIQQQGBSDOWNP-UHFFFAOYSA-N 2-dodecylbenzenesulfonic acid Chemical compound CCCCCCCCCCCCC1=CC=CC=C1S(O)(=O)=O WBIQQQGBSDOWNP-UHFFFAOYSA-N 0.000 claims description 2
- LIFHMKCDDVTICL-UHFFFAOYSA-N 6-(chloromethyl)phenanthridine Chemical compound C1=CC=C2C(CCl)=NC3=CC=CC=C3C2=C1 LIFHMKCDDVTICL-UHFFFAOYSA-N 0.000 claims description 2
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- 125000005227 alkyl sulfonate group Chemical group 0.000 claims description 2
- BTBJBAZGXNKLQC-UHFFFAOYSA-N ammonium lauryl sulfate Chemical compound [NH4+].CCCCCCCCCCCCOS([O-])(=O)=O BTBJBAZGXNKLQC-UHFFFAOYSA-N 0.000 claims description 2
- 229940060296 dodecylbenzenesulfonic acid Drugs 0.000 claims description 2
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- PYSRRFNXTXNWCD-UHFFFAOYSA-N 3-(2-phenylethenyl)furan-2,5-dione Chemical compound O=C1OC(=O)C(C=CC=2C=CC=CC=2)=C1 PYSRRFNXTXNWCD-UHFFFAOYSA-N 0.000 description 1
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- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 1
- 238000012921 fluorescence analysis Methods 0.000 description 1
- RPOCFUQMSVZQLH-UHFFFAOYSA-N furan-2,5-dione;2-methylprop-1-ene Chemical compound CC(C)=C.O=C1OC(=O)C=C1 RPOCFUQMSVZQLH-UHFFFAOYSA-N 0.000 description 1
- COBLIZNSZVKDMR-UHFFFAOYSA-N furan-2,5-dione;octadec-1-ene Chemical compound O=C1OC(=O)C=C1.CCCCCCCCCCCCCCCCC=C COBLIZNSZVKDMR-UHFFFAOYSA-N 0.000 description 1
- 150000002433 hydrophilic molecules Chemical class 0.000 description 1
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- 231100000956 nontoxicity Toxicity 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
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- 238000006467 substitution reaction Methods 0.000 description 1
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- 125000003698 tetramethyl group Chemical group [H]C([H])([H])* 0.000 description 1
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Abstract
The invention discloses a preparation method of aggregation-induced emission microsphere with core-shell structure, which comprises dissolving aggregation-induced emission material and amphiphilic polymer as raw materials in good solvent, adding water solution with surfactant, making the solution into microemulsion, removing good solvent in the microemulsion, centrifuging, redissolving the obtained precipitate in alkaline water, hydrolyzing, centrifuging, cleaning, redissolving the cleaned precipitate in buffer solution, and obtaining aggregation-induced emission microsphere with core-shell structure; the raw materials also comprise fat-soluble colored dye, and the aggregation-induced emission microsphere with core-shell structure, colorimetric and fluorescent functions is prepared. The method has the advantages of simple process, no need of complex instruments, short time, low reagent cost, high stability and good reproducibility, and has good commercialization prospect; and the prepared aggregation-induced emission microsphere has the advantages of complete appearance, good dispersibility, narrow particle size distribution, strong fluorescence signal and controllable particle size.
Description
Technical Field
The invention belongs to the technical fields of nano material synthesis, food safety detection and biomedical inspection, and particularly relates to a preparation method of aggregation-induced emission microspheres with core-shell structures.
Background
The fluorescence analysis technology is widely applied to the fields of food safety rapid detection, biomedical inspection and the like due to the advantages of convenience in detection, rapid sensitivity, real-time detection and the like, and plays a key role in guaranteeing food safety and disease diagnosis and treatment. Because the traditional organic fluorescent dye has a rigid plane structure, the fluorescence of the traditional organic fluorescent dye is greatly weakened or even quenched in a high concentration or aggregation state, so that the application of the traditional organic fluorescent dye in the field of high-sensitivity analysis is severely restricted.
Until 2001, the group of the academy of sciences and sciences at hong Kong university Tang Benzhong first proposed the concept of "aggregation-induced emission (AIE)", which opened a new way to solve this problem. Compared with the traditional aggregation quenching organic fluorescent dye, the fluorescent dye with aggregation-induced emission property has weak light emission even difficult to observe in a dilute solution state, but can emit bright fluorescence when aggregation occurs in the solution or in a solid state. Aggregation-induced emission type fluorescent dyes have become a research hotspot in the fluorescence field due to their unique properties of solid state emission.
The existing synthesis methods of aggregation-induced emission microspheres mainly comprise an emulsion polymerization method, a self-assembly method and a microemulsion method. Emulsion polymerization (CN 104628923A, CN104628924 a) allows the synthesis of structurally stable polymer microspheres, but with the presence of an initiator initiated polymerization, thus resulting in a longer synthesis period (> 2 days). The self-assembly method (CN 113788795A) connects the aggregation-inducing luminescent material and the hydrophilic compound by chemical synthesis to form an amphiphilic chemical with one hydrophilic end and the other hydrophobic end. In microsphere synthesis, no initiator is needed due to hydrophilic-hydrophobic interactions, and spontaneous balling is achieved. Because AIE molecules and hydrophilic chemicals need to be connected through chemical synthesis in advance, the pretreatment process for synthesizing the microsphere by a self-assembly method is complex, and the universality is low. And its dye loading efficiency is low due to the limitation of the structure of the compound (the fixation of the dye and the polymer is 1:1). The microemulsion method combines the advantages of simple synthesis process and short synthesis time of the self-assembly method, and can synthesize the aggregation-induced emission microsphere in a short time by a simple process. For example, patent CN109749326B prepares tetramethyl tetrastilbene fluorescent microsphere by microemulsion method, but contains organic glass and polymethyl methacrylate, but the introduction of polymethyl methacrylate can lead to the reduction of the loading of fluorescent dye, resulting in the optical performance of fluorescent microsphere not high. By removing polymethyl methacrylate in the organic phase, the dye loading of the microspheres can be increased. Therefore, the invention aims to remove the organic glass, adopts the amphiphilic polymer and synthesizes the AIE fluorescent microsphere by a microemulsion method. Because AIE dyes are rich in benzene rings, and have a polarity difference from polymers, phase separation occurs during the volatilization of the organic phase, forming a typical core-shell structure. The outer polymer shell not only provides various functional groups, but also protects the fluorescent groups, thereby improving the environmental tolerance. Has bright prospect in application.
Disclosure of Invention
Aiming at the defects and the problems in the prior art, the invention aims to provide a preparation method of aggregation-induced emission microspheres with core-shell structures. The invention provides a method for stably preparing 50-1000nm aggregation-induced emission microsphere, and the prepared aggregation-induced emission microsphere has the characteristics of high fluorescence intensity, narrow particle size distribution, high sphericity and the like.
Because the hydrophobicity of the aggregation-induced emission material is far higher than that of the amphiphilic polymer, phase separation can occur when the good solvent is removed, the aggregation-induced emission material is contracted inwards to form a nucleus, the amphiphilic polymer is in a shell outside, and the prepared aggregation-induced emission microsphere has a vivid core-shell structure. The amphiphilic polymer shell plays a role in protecting the AIE inner core, isolates the influence of the external extreme environment on the internal fluorescent groups, and the prepared fluorescent microsphere has the advantages of higher stability and better environmental tolerance. As shown in fig. 2, the synthesized aggregation-induced emission microsphere has a core-shell structure.
The invention is realized by the following technical scheme:
The invention provides a preparation method of aggregation-induced emission microsphere with core-shell structure, which comprises the steps of dissolving aggregation-induced emission material and amphiphilic polymer as raw materials in a good solvent, adding aqueous solution with surfactant, preparing the solution into microemulsion, removing the good solvent in the microemulsion, dissolving the obtained precipitate again after centrifugal separation, centrifugally cleaning after hydrolysis, and dissolving the cleaned precipitate in buffer solution again to obtain the aggregation-induced emission microsphere with core-shell structure.
Further, the raw materials also comprise fat-soluble colored dye, and the aggregation-induced emission microsphere with core-shell structure, colorimetric and fluorescent functions is prepared.
Further, the aggregation-induced emission material is selected from any one of the formulas (I), (II) and (III) to respectively prepare the aggregation-induced emission microsphere with red, yellow and green emission; the aggregation-induced emission material is respectively selected from more than two of the formulas (I), (II) and (III) and mixed and doped according to different proportions to prepare aggregation-induced emission microspheres with various colors;
Further, the fat-soluble colored dye comprises any one or more of red, green, blue and black, and the final concentration of the fat-soluble colored dye after being dissolved in a good solvent is 1-200mg/mL.
Further, the good solvent comprises any one of dichloromethane, chloroform, dichloroethane, cyclohexane, cyclopentane and n-butanol; the amphiphilic polymer comprises any one of styrene maleic anhydride polymer, isobutylene maleic anhydride polymer, octadecene maleic anhydride polymer and lactic acid ethylene glycol copolymer; the surfactant comprises any one of ammonium dodecyl sulfate, sodium dodecyl sulfate, dodecylbenzene sulfonic acid, triethanolamine lauryl sulfate and sodium secondary alkyl sulfonate.
Further, the final concentration of the aggregation-induced emission material after being dissolved in a good solvent is 1-200mg/mL; the final concentration of the amphiphilic polymer after being dissolved in a good solvent is 1-500mg/mL; the final concentration of the aqueous surfactant solution is 0.5-10mg/mL.
Further, the microemulsion is prepared at 0-35 ℃, the emulsifying time is 0.5-10 minutes, and the emulsifying power is 50-500W; the alkaline water is sodium hydroxide aqueous solution with pH=9-12, and the hydrolysis time is 24 hours; removing good solvent in the microemulsion by rotary evaporation, wherein the rotary evaporation condition is that the vacuum degree is more than 0.085MPa, and the time is 10-60 minutes; the centrifugal speed is 3000-16000rpm after the good solvent in the microemulsion is removed, and the time is 10-30 minutes.
Compared with the prior art, the invention has the beneficial effects that:
1. The aggregation-induced fluorescence dye organic molecule adopted by the invention has the advantages of high quantum efficiency, no toxicity or harm, easy synthesis, rich structure, simple process, no need of complex instruments, short time, low reagent cost, high stability and good reproducibility, and has better commercialization prospect; and the prepared aggregation-induced emission microsphere has the advantages of complete appearance, good dispersibility, narrow particle size distribution, strong fluorescence signal and controllable particle size.
2. Due to the interaction of hydrophilicity and hydrophobicity, an AIE inner core and a polymer outer shell are spontaneously formed, the polymer outer shell not only provides protection for the AIE inner core, but also can provide various functional groups such as carboxyl, phenylboronic acid and the like through modification; the aggregation-induced emission microsphere with different particle sizes can be prepared by adjusting the volume of the oil phase, the oil-water ratio, the concentration of the surfactant, the emulsifying power and the emulsifying temperature; the aggregation-induced emission microsphere prepared by the invention has good light-emitting performance, and the light-emitting intensity is improved by 64 times compared with that of the traditional FITC fluorescent microsphere.
3. The invention can also prepare the aggregation-induced emission microsphere with colorimetric and fluorescent functions, and can simultaneously provide naked eye interpretation and high-sensitivity quantitative detection capabilities.
Drawings
FIG. 1 is a flow chart showing the preparation of the multicolor aggregation-induced emission microsphere with core-shell structure according to example 1 of the present invention.
FIG. 2 shows the structural formula of a red aggregation-induced emission material according to the present invention.
FIG. 3 shows the structural formula of a yellow aggregation-induced emission material according to the present invention.
FIG. 4 shows the structural formula of a green aggregation-induced emission material according to the present invention.
FIG. 5 is a scanning electron microscope image of aggregation-induced emission microspheres of different particle sizes synthesized in example 1 of the present invention, wherein A) 250nm, B) 350nm and C) 450nm.
FIG. 6 is a flow chart showing the preparation of the aggregation-induced emission microsphere with core-shell structure and colorimetric fluorescence dual function according to example 2 of the present invention.
FIG. 7 is a transmission electron microscope image of aggregation-induced emission microspheres synthesized in example 2 of the present invention.
Detailed Description
The invention will be further described with reference to the accompanying drawings.
Example 1 preparation of multicolor aggregation-induced emission microsphere having core-shell Structure
The preparation flow chart of the multicolor aggregation-induced emission microsphere with the core-shell structure is shown in figure 1, and the specific process is as follows:
1) Preparation of aggregation-induced emission microspheres: dissolving aggregation-induced emission dye (selected from any one or more of the figures 2, 3 and 4) and polymaleic anhydride octadecene (PMAO) in chloroform (forming an oil phase), wherein the concentration of the aggregation-induced emission dye after dissolving in the chloroform is 10-100 mg/mL, the concentration of the PMAO after dissolving in the chloroform is 10-500 mg/mL, and then adding sodium dodecyl sulfate solution (water phase) with the mass concentration of 0.5-10%, wherein the volume ratio of the water phase to the oil phase is 1:4, uniformly mixing, and oscillating and ultrasonic to form coarse emulsion.
Crushing the solution by using a cell breaker, performing ultrasonic emulsification at 25 ℃ for 2 minutes (9.9 seconds on; 5.0 seconds off) with the emulsification power of 150W, performing rotary evaporation on the emulsion at 0.085MPa for 10 minutes, removing chloroform, centrifuging at 13500 r for 20 minutes to remove supernatant, performing ultra-pure water washing on the precipitate for three times, redissolving the precipitate in ultrapure water or alkaline water with pH of 10, performing centrifugal washing on the precipitate for 3 times, and obtaining the aggregation-induced emission microspheres, and re-dissolving the aggregation-induced emission microspheres in a buffer solution.
Description: FIGS. 2,3 and 4 are structural formulas of red, yellow and green aggregation-inducing emission materials, respectively, in which various combinations thereof may form other colors according to a certain ratio, and red+yellow may form orange; the emission wavelengths of AIE formulas 1, 4, 6 in FIG. 2 are 590-630nm, and the emission wavelengths of AIE formulas 8-11 in FIG. 3 are 550-580nm; the emission wavelengths of AIE formulas 12, 14, 15 in FIG. 4 are 500-540nm.
FIG. 5 is a scanning electron microscope image of aggregation-induced emission microspheres of different particle sizes synthesized in this example, wherein A) 250nm, B) 350nm and C) 450nm.
Example 2 preparation of colorimetric and fluorescent bifunctional aggregation-induced emission microsphere with core-shell Structure
As shown in fig. 6, the preparation process of the aggregation-induced emission microsphere with core-shell structure and dual functions of colorimetry and fluorescence is as follows:
Red or yellow or green aggregation-induced emission material, fat-soluble black dye and polymaleic anhydride octadecene are respectively weighed and dissolved in chloroform, and the final concentrations are respectively 25mg/mL, 25mg/mL and 25mg/mL. An aqueous solution of 10mg/mL sodium dodecyl sulfate was prepared. The ratio of the aqueous phase to the oil phase was 100. Mu.l: 400 microliters, shaking ultrasound, forming a coarse emulsion. The emulsion was performed at 25℃for 2 minutes (9.9 seconds on; 5.0 seconds off) using a cell disrupter with an emulsifying power of 150W. The obtained microemulsion is redissolved in 1mL of ultrapure water, then transferred into a rotary steaming bottle, rotary steamed for 10 minutes under the pressure of 0.085MPa, and the chloroform in the microemulsion is removed. Centrifugation was carried out at 13500 rpm for 20 min, the supernatant was removed, redissolved in 1mL of alkaline water at ph=10 and hydrolysed for 24 h. The mixture was centrifuged and washed 3 times with ultrapure water. Finally, the aggregation-induced emission microsphere is redissolved in ultrapure water to prepare the colorimetric and fluorescent dual-functional aggregation-induced emission microsphere with a core-shell structure, and a transmission electron microscope diagram of the aggregation-induced emission microsphere is shown in figure 7.
The prepared aggregation-induced emission microsphere has colorimetric and fluorescent dual-signal output; the prepared immunochromatographic test strip has the characteristics of naked eye interpretation and high-sensitivity accurate quantitative detection.
The preparation process of the PCT colorimetric and fluorescent double-signal output immunochromatographic test strip is as follows:
2mL of PB6.0 was placed in a sample bottle, and 0.5mg of aggregation-induced emission microsphere was added. 50 μg of PCT-labeled antibody was added and incubated for 30 minutes. EDC was added to the flask in 3 portions, 25 μg each, and incubated for 40 minutes after each addition. 40mg of skim milk was added for blocking and incubated for 1 hour. The blocked probe was centrifuged at 15000rpm for 15 minutes, the supernatant was removed, and the probe complex solution was added for preservation. The synthesized aggregation-induced emission probe was sprayed onto the conjugate pad in a spray amount of 3. Mu.l/cm. And (3) spraying 1.8mg/mL of PCT (PCT) labeled antibody and 1.0mg/mL of goat anti-mouse antibody on a test line and a quality control line on the nitrocellulose membrane respectively, wherein the spraying amount is 0.95 mu L/cm. And respectively sticking a nitrocellulose membrane, a combination pad sprayed with a probe, a sample pad and absorbent paper on the PVC bottom plate. And then stored in a drying cabinet.
The foregoing description of the preferred embodiments of the present invention has been presented only in terms of those specific and detailed descriptions, and is not, therefore, to be construed as limiting the scope of the invention. It should be noted that modifications, improvements and substitutions can be made by those skilled in the art without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.
Claims (2)
1. A preparation method of aggregation-induced emission microspheres with core-shell structures is characterized by comprising the following steps: dissolving aggregation-induced emission material and amphiphilic polymer as raw materials in a good solvent, adding an aqueous solution with a surfactant, preparing the solution into microemulsion, removing the good solvent in the microemulsion, dissolving the obtained precipitate in pure water or alkaline water again after centrifugal separation, centrifugally cleaning after hydrolysis, and dissolving the cleaned precipitate in a buffer solution again to prepare the aggregation-induced emission microsphere with a core-shell structure;
The raw materials also comprise fat-soluble colored dye, and the aggregation-induced emission microsphere with core-shell structure, colorimetric and fluorescent functions is prepared;
The aggregation-induced emission material is selected from any one of a formula (I), a formula (II) and a formula (III) to respectively prepare aggregation-induced emission microspheres with red, yellow and green emission; the aggregation-induced emission material is prepared by mixing and doping more than two of the materials in the formulas (I), (II) and (III) according to different proportions to obtain aggregation-induced emission microspheres with various colors;
，
(I)
，
(II)
，
(III)
The fat-soluble colored dye is selected from one or more of red, green, blue and black, and the final concentration of the fat-soluble colored dye after being dissolved in a good solvent is 1-200mg/mL;
The good solvent is selected from any one of dichloromethane, chloroform, dichloroethane, cyclohexane, cyclopentane and n-butanol; the amphiphilic polymer adopts polymaleic anhydride octadecene; the surfactant is selected from any one of ammonium dodecyl sulfate, sodium dodecyl sulfate, dodecylbenzene sulfonic acid, triethanolamine lauryl sulfate and sodium secondary alkyl sulfonate;
The final concentration of the aggregation-induced emission material after being dissolved in a good solvent is 1-200mg/mL; the final concentration of the amphiphilic polymer after being dissolved in a good solvent is 1-500mg/mL; the final concentration of the aqueous surfactant solution is 0.5-10mg/mL.
2. The method for preparing the aggregation-induced emission microsphere with the core-shell structure according to claim 1, wherein the method comprises the following steps: the microemulsion is prepared at 0-35 ℃, the emulsifying time is 0.5-10 minutes, and the emulsifying power is 50-500W;
the alkaline water is sodium hydroxide aqueous solution with pH=9-12, and the hydrolysis time is 24 hours;
removing good solvent in the microemulsion by rotary evaporation, wherein the rotary evaporation condition is that the vacuum degree is more than 0.085MPa, and the time is 10-60 minutes;
the centrifugal speed is 3000-16000rpm after the good solvent in the microemulsion is removed, and the time is 10-30 minutes.
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