阅读说明：本技术 一种多孔高荧光聚丙烯酸酯微球及其制备方法 (Porous high-fluorescence polyacrylate microsphere and preparation method thereof ) 是由 詹伟东 邓锋 孙宾 杨炎 陈珈 李俊 于 2020-07-09 设计创作，主要内容包括：本发明涉及一种多孔高荧光聚丙烯酸酯微球及其制备方法,制备具体为：(1)将乳化剂和去离子水混合形成体系I；(2)将丙烯酸甲酯和1,7-乙烯基-苝酰亚胺衍生物溶于有机溶剂中再加入到体系I中得到体系II；(3)在体系II中加入过硫酸钾引发聚合得到聚丙烯酸酯分散液；进而得到多孔高荧光聚丙烯酸酯微球；该微球是以1,7-乙烯基-苝酰亚胺衍生物作为交联剂制得的多孔高荧光聚丙烯酸酯微球；在440～460nm波长的激发下,产生630～645nm的1,7-乙烯基-苝酰亚胺衍生物特征荧光发射。本发明的方法简单、安全；制备得到微球应用方便,受到基体限制较小,调控添加量方便,在不同激发波长下可以发射特定的荧光,具有可识别性能。(The invention relates to a porous high-fluorescence polyacrylate microsphere and a preparation method thereof, and the preparation method specifically comprises the following steps: (1) mixing an emulsifier and deionized water to form a system I; (2) dissolving methyl acrylate and 1, 7-vinyl-perylene bisimide derivative in an organic solvent, and adding the solution into the system I to obtain a system II; (3) adding potassium persulfate into the system II to initiate polymerization to obtain polyacrylate dispersion liquid; thereby obtaining porous high-fluorescence polyacrylate microspheres; the microsphere is a porous high-fluorescence polyacrylate microsphere prepared by using a 1, 7-vinyl-perylene bisimide derivative as a cross-linking agent; under the excitation of the wavelength of 440-460 nm, the characteristic fluorescence emission of the 630-645 nm 1, 7-vinyl-perylene bisimide derivative is generated. The method is simple and safe; the prepared microsphere is convenient to apply, is less limited by a matrix, is convenient to adjust and control the addition amount, can emit specific fluorescence under different excitation wavelengths, and has identifiable performance.)

1. A porous high-fluorescence polyacrylate microsphere is characterized in that: the porous high-fluorescence polyacrylate microsphere is prepared by taking a 1, 7-vinyl-perylene bisimide derivative as a cross-linking agent; the porous high-fluorescence polyacrylate microsphere generates 630-645 nm characteristic fluorescence emission of the 1, 7-vinyl-perylene bisimide derivative under the excitation wavelength of 440-460 nm;

the 1, 7-vinyl-perylene bisimide derivative is perylene bisimide with a substituent group with an ethylene group at gulf position (1,7 positions) and a bulky substituent group at an imide position.

2. The porous high-fluorescence polyacrylate microsphere of claim 1, wherein the bulky substituent is a sesqui-cage siloxane or a long alkyl chain with a side chain;

the silsesquioxane is

the long alkyl chain with side chain is

WhereinIndicates that the linking position of the chemical bond is an N atom in an imide structure;

the substituent of the ethylene group is an alkyl chain with an ethylene group at the end group, and the alkyl chain is an alkyl chain with less than six carbons.

3. The porous high-fluorescence polyacrylate microsphere of claim 1, wherein the molar ratio of the 1, 7-vinyl-perylene imide derivative to the acrylate structural unit is 14-21.5: 125.

4. The porous high-fluorescence polyacrylate microsphere of claim 1, wherein the fluorescence quantum yield of the porous high-fluorescence polyacrylate microsphere is 95-99%.

5. The porous high-fluorescence polyacrylate microsphere of claim 1, wherein the average diameter of the porous high-fluorescence polyacrylate microsphere is 150-300 nm, the average pore diameter is 10-30 nm, and the porosity is 35-55%.

6. The preparation method of the porous high-fluorescence polyacrylate microspheres as claimed in any one of claims 1 to 5, which is characterized by comprising the following steps:

(1) mixing an emulsifier and deionized water at a temperature T1 to form a system I;

(2) firstly, dissolving methyl acrylate and 1, 7-vinyl-perylene bisimide derivatives in an organic solvent, adding the mixture into a system I, and mixing at a temperature of T2 to obtain a system II;

(3) adding potassium persulfate into the system II to initiate polymerization to obtain polyacrylate dispersion liquid; and cooling the polyacrylate dispersion liquid to room temperature, filtering, washing and drying to obtain the porous high-fluorescence polyacrylate microspheres.

7. The method for preparing porous high-fluorescence polyacrylate microspheres according to claim 6, wherein the emulsifier is potassium laurate, sodium dodecyl sulfate or sodium dioctyl sulfosuccinate.

8. The method for preparing porous polyacrylate microsphere with high fluorescence according to claim 6, wherein the organic solvent is toluene or xylene.

9. The preparation method of the porous high-fluorescence polyacrylate microsphere of claim 6, wherein in the step (1), T1 is 35-55 ℃;

in the system II in the step (2), the content of the emulsifier is 0.4-0.7 wt%, the content of the methyl acrylate is 4-6 wt%, the content of the 1, 7-vinyl-perylene imide derivative is 9-15 wt%, and the content of the organic solvent is 6-10 wt%; t2 is 75-95 ℃;

in the step (3), the mass ratio of the addition amount of the potassium persulfate to the methyl acrylate is 0.0024-0.006: 1; the polymerization time is 4-8 h, and the polymerization temperature is 75-95 ℃.

Technical Field

The invention belongs to the technical field of fluorescent microspheres, and relates to a porous high-fluorescence polyacrylate microsphere and a preparation method thereof.

Background

The fluorescent microspheres are solid particles which have particle diameters ranging from nanometer to micron, are loaded with fluorescent substances and can excite fluorescence under the stimulation of external energy. Due to the unique properties and excellent performance of the fluorescent microspheres, the fluorescent microspheres are widely applied to the fields of cell surface antigen detection, cell phagocytic function detection, lesion tracers, blood flow analysis, diagnostic reagents and the like in the aspect of biological analysis.

Patent CN201310219573.9 relates to a method for preparing polymer fluorescent microspheres, which comprises the following steps: firstly, preparing polymer fluorescent microsphere liquid drops: dissolving a first polymer in a solvent, adding a fluorescent material, and uniformly stirring by magnetic force to obtain a discontinuous phase; respectively filling the discontinuous phase and the continuous phase into injectors connected with a discontinuous phase inlet and a continuous phase inlet of the microfluid device, adjusting the flow rate of two-phase solution by a micro-sampling pump, obtaining fluorescent microsphere liquid drops at a microfluid outlet, and collecting the fluorescent microsphere liquid drops in a fluorescent microsphere receiving device; step two, preparing the polymer fluorescent microspheres: and drying the fluorescent microsphere liquid drops until the solvent is completely volatilized, washing with a detergent, and cleaning the continuous phase solution to obtain the polymer fluorescent microsphere finally. The method has the advantages of simple equipment and convenient operation, and can adjust the particle size of the microspheres by adjusting the flow rates of the continuous phase and the discontinuous phase, so that the prepared microspheres have high particle size uniformity.

Patent CN201910160304.7 is a manufacturing method for providing a fluorescent microsphere test piece, which comprises a raw material containing area, wherein a fluorescent microsphere solution is dropped into the raw material containing area to naturally level and dry to form a layer of fluorescent microspheres, resin is dropped onto the fluorescent microspheres, a cover glass is covered to fill the encapsulating space formed in the raw material containing area with resin without gaps, and the cover glass is peeled off after the resin is cured to obtain the fluorescent microsphere test piece. In the method, the fluorescent microsphere solution is dripped into the raw material accommodating area to naturally level and air-dry to form a layer of tiled fluorescent microspheres, so that the fluorescent microspheres are prevented from being stacked and piled on any longitudinal arrangement, and accurate detection results can be obtained; the thickness of the finally prepared fluorescent microsphere test piece is controlled by the measuring scale, so that fluorescent microsphere test pieces with different thicknesses can be obtained according to requirements.

Patent CN201611143884.1 relates to a method for preparing nano fluorescent microspheres, which comprises the following steps: preparation of fluorescent dye solution: dissolving a fluorescent dye in an organic solvent, and diluting to obtain a fluorescent dye solution; preparing the nano fluorescent microspheres: adding a comonomer, an initiator, a cross-linking agent, a dispersant and a fluorescent dye solution into a reaction vessel, stirring and dispersing uniformly, and reacting to prepare a nano fluorescent microsphere solution; dialysis of the nano fluorescent microsphere solution: and (3) dialyzing the nano fluorescent microsphere solution to remove unreacted monomers and impurities. The beneficial effects are as follows: NIPAM and DMC both have good water solubility, the polymer microsphere prepared by using the same also has good water solubility, and the NIPAM is added into the material for synthesizing the microsphere, so that a certain temperature-sensitive property can be brought to the product; the fluorescent microspheres prepared by copolymerizing and embedding the fluorescent dye by using the two monomers have good biocompatibility, can grow together with cells, and does not influence the activity of the cells.

The fluorescent microspheres obtained by the above patents have respective characteristics, but the fluorescence emission intensity of the obtained microspheres is not outstanding enough, in the fluorescent microspheres, the microspheres with a porous structure are concerned, the porous polymer microspheres are concerned due to the unique structure thereof, the porous structure can increase the specific surface area of a carrier on one hand, so that more active components can be fixed, and on the other hand, channels can help reactants and products to diffuse therein, so that the reaction efficiency is improved, and the fluorescent microspheres have wide application prospects in the aspects of immobilized enzymes, targeted drugs, immunoassay, cell separation, high-grade cosmetics, environment-friendly high-efficiency catalysts and the like. Many methods for making the polymer have pores, such as solvent pore-forming method in suspension polymerization, template pore-forming method, etc., and are currently used.

Disclosure of Invention

The invention aims to solve the problem that the fluorescence emission intensity of fluorescent microspheres in the prior art is not high enough, and provides porous high-fluorescence polyacrylate microspheres and a preparation method thereof.

One of the purposes is to provide a porous high-fluorescence polyacrylate microsphere, which is prepared by taking 1, 7-vinyl-perylene bisimide derivatives as a cross-linking agent; the porous high-fluorescence polyacrylate microsphere generates 630-645 nm characteristic fluorescence emission of the 1, 7-vinyl-perylene bisimide derivative under the excitation wavelength of 440-460 nm;

the second purpose is to provide a preparation method of the porous high-fluorescence polyacrylate microsphere, which comprises the following steps: (1) mixing an emulsifier and deionized water to form a system I; (2) dissolving methyl acrylate and 1, 7-vinyl-perylene bisimide derivative in an organic solvent, and adding the solution into the system I to obtain a system II; (3) adding potassium persulfate into the system II to initiate polymerization to obtain polyacrylate dispersion liquid; and cooling the polyacrylate dispersion liquid to room temperature, filtering, washing and drying to obtain the porous high-fluorescence polyacrylate microspheres.

In order to achieve the purpose, the invention adopts the following scheme:

a porous high-fluorescence polyacrylate microsphere is prepared by taking 1, 7-vinyl-perylene bisimide derivatives as a cross-linking agent; the porous high-fluorescence polyacrylate microsphere generates 630-645 nm characteristic fluorescence emission of the 1, 7-vinyl-perylene bisimide derivative under the excitation wavelength of 440-460 nm, and the color is orange yellow;

the 1, 7-vinyl-perylene bisimide derivative is perylene bisimide with a substituent group with an ethylene group at gulf position (1,7 positions) and a bulky substituent group at an imide position.

The 1, 7-vinyl-perylene imide derivatives in the present invention function as: the 1, 7-vinyl-perylene imide derivative with the bulky imide site substituent can enable the 1, 7-vinyl-perylene imide derivative to have great steric hindrance when being aggregated through pi-pi interaction, and can be more easily existed in a system in a monomolecular state in the presence of a solvent. Finally, the 1, 7-vinyl-perylene imide derivative can be used as a cross-linking agent to enter the polyacrylate microsphere in a monomolecular state. The 1, 7-vinyl-perylene imide derivative is also a fluorescent molecule, and the 1, 7-vinyl-perylene imide derivative generates fluorescence quenching when being aggregated through pi-pi interaction, so that the fluorescence quantum yield is reduced, and the related fluorescence performance is reduced. According to the invention, the 1, 7-vinyl-perylene bisimide derivative enters a system in a monomolecular state, so that the aggregation of the derivative is effectively avoided, the occurrence of fluorescence quenching is avoided, and the good fluorescence property of the 1, 7-vinyl-perylene bisimide derivative is maintained.

As a preferred technical scheme:

the porous high-fluorescence polyacrylate microsphere is characterized in that the bulky substituent is sesqui-cage siloxane or long alkyl chain with side chain;

the silsesquioxane is

R is isobutyl or isooctyl;

the long alkyl chain with side chain is

WhereinIndicates that the linking position of the chemical bond is an N atom in an imide structure;

the substituent of the ethylene group is an alkyl chain with an ethylene group at the end group, and the alkyl chain is an alkyl chain with less than six carbons.

According to the porous high-fluorescence polyacrylate microsphere, the molar ratio of the 1, 7-vinyl-perylene bisimide derivative to the acrylate structural unit is 14-21.5: 125.

According to the porous high-fluorescence polyacrylate microsphere, the fluorescence quantum yield of the porous high-fluorescence polyacrylate microsphere is 95-99%, the fluorescence emission peak of the microsphere is kept at a low wavelength, and the red shift of a monomolecular state in a solution is small; fluorescence emission at low wavelengths is maintained and small red-shifted relative to unimolecular states indicates that no significant aggregation of the molecules occurs and fluorescence is not quenched by aggregation, so high fluorescence quantum yields can be maintained.

The porous high-fluorescence polyacrylate microsphere has the advantages that the average diameter is 150-300 nm, the particle size is uniform, the average pore diameter is 10-30 nm, and the porosity is 35-55%.

The preparation method of the porous high-fluorescence polyacrylate microsphere comprises the following steps:

(1) mixing an emulsifier and deionized water at a temperature T1 to form a system I;

(2) firstly, dissolving methyl acrylate and the 1, 7-vinyl-perylene imide derivative in an organic solvent, adding the methyl acrylate and the 1, 7-vinyl-perylene imide derivative into the system I, and mixing at a temperature of T2 to obtain a system II, wherein the organic solvent is favorable for dispersing the 1, 7-vinyl-perylene imide derivative without generating obvious aggregation and is favorable for subsequently entering the polyacrylate microspheres in a monomolecular state;

(3) stirring the system II for a certain time, and adding potassium persulfate into the system II to initiate polymerization to obtain a polyacrylate dispersion liquid; and cooling the polyacrylate dispersion liquid to room temperature (23 +/-2 ℃), filtering, washing and drying to obtain the porous high-fluorescence polyacrylate microspheres (solid powder).

According to the preparation method of the porous high-fluorescence polyacrylate microsphere, the emulsifier is potassium laurate, sodium dodecyl sulfate or sodium dioctyl sulfosuccinate.

According to the preparation method of the porous high-fluorescence polyacrylate microsphere, the organic solvent is toluene or xylene.

The preparation method of the porous high-fluorescence polyacrylate microsphere comprises the following steps of (1), wherein T1 is 35-55 ℃, and the mixing time is 3-8 min;

in the system II in the step (2), the content of the emulsifier is 0.4-0.7 wt%, the content of the methyl acrylate is 4-6 wt%, the content of the 1, 7-vinyl-perylene imide derivative is 9-15 wt%, and the content of the organic solvent is 6-10 wt%; t2 is 75-95 ℃;

in the step (3), the mass ratio of the addition amount of the potassium persulfate to the methyl acrylate is 0.0024-0.006: 1, the stirring speed is 300-500 r/min, and the stirring time is 15-35 min; the polymerization time is 4-8 h, and the polymerization temperature is 75-95 ℃; the drying temperature is 90-140 ℃.

The principle of the invention is as follows:

the high-fluorescence polyacrylate microsphere is prepared by introducing fluorescence molecules into a polyacrylate molecular chain structure through double bond reaction, but the fluorescence monomer of the copolymerization method is copolymerized by adopting molecules with single functionality, is connected into a high molecular chain, and only plays a role of a fluorescent substance in the material. The 1, 7-vinyl-perylene bisimide derivative has two functionalities, can connect two macromolecular chains in the preparation process of the polyacrylate microsphere, plays a role in crosslinking in the polyacrylate microsphere and also plays a role in a fluorescent substance. In the preparation process, a good solvent of the 1, 7-vinyl-perylene bisimide derivative is added, so that the molecule enters a molecular chain in a monomolecular state, and fluorescence quenching caused by aggregation among molecules is avoided, so that the prepared microsphere has high fluorescence performance. The porous structure of the high-fluorescence polyacrylate microsphere is beneficial for more fluorescent substances to receive light, so that the fluorescent substances in the microsphere can also receive exciting light to participate in fluorescence emission.

Has the advantages that:

(1) the porous high-fluorescence polyacrylate microsphere can emit specific fluorescence under different excitation wavelengths, and has identifiable performance;

(2) the preparation method of the porous high-fluorescence polyacrylate microsphere is simple and safe; the prepared microspheres are convenient to apply, less limited by a matrix and convenient to adjust and control the addition amount.

Detailed Description

The invention will be further illustrated with reference to specific embodiments. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Further, it should be understood that various changes or modifications of the present invention may be made by those skilled in the art after reading the teaching of the present invention, and such equivalents may fall within the scope of the present invention as defined in the appended claims.