CN-117225318-B - Preparation method of carboxylated porous microspheres

Abstract

The application relates to the technical field of microspheres, in particular to a preparation method of carboxylated porous microspheres, which comprises the steps of adding a biocompatible polymer, an acrylic copolymer and an oil-soluble emulsifier into an organic solvent for dissolution to prepare a disperse phase; adding water-soluble emulsifying agent and/or protective colloid into water to prepare continuous phase, mixing disperse phase and continuous phase to obtain emulsion, continuously stirring until precipitation appears, collecting precipitation, cleaning and drying to obtain the carboxylated porous microsphere. Aeration or addition of a cross-linking agent may also be performed during the continuous agitation phase. The carboxylated porous microspheres have good fluidity and good nucleic acid extraction and purification effects.

Inventors

• CHEN ZHIJIAN
• LIU FENG
• LIN ZHIKENG

Assignees

• 厦门为正生物科技股份有限公司

Dates

Publication Date

20260508

Application Date

20230926

Claims (10)

1. 1. The preparation method of the carboxylated porous microspheres is characterized by comprising the following steps: S1, adding a biocompatible polymer, an acrylic acid copolymer and an oil-soluble emulsifier into an organic solvent for dissolution to prepare a disperse phase, wherein the biocompatible polymer is selected from polylactic acid or polylactic acid-glycolic acid copolymer; S2, adding a water-soluble emulsifier and/or a protective colloid into water to prepare a continuous phase; And S3, mixing the disperse phase in the step S1 and the continuous phase in the step S2 to obtain emulsion, continuously stirring until precipitation appears, collecting the precipitation, cleaning, and drying to obtain the carboxylated porous microspheres.
2. 2. The method of preparing carboxylated porous microspheres according to claim 1, wherein the weight ratio of the biocompatible polymer to the acrylic copolymer in step S1 is 1:0.01-0.2.
3. 3. The method for preparing carboxylated porous microspheres according to claim 1, wherein the weight ratio of acrylic acid in the acrylic copolymer in step S1 is 1 to 50%.
4. 4. The method for preparing carboxylated porous microspheres according to claim 1, wherein the boiling point of the organic solvent in step S1 is not more than 120 ℃ at 1 atmosphere.
5. 5. The method for preparing carboxylated porous microspheres according to claim 1, wherein the concentration of the dispersed phase in step S1 is 1 to 60wt%.
6. 6. The method of preparing carboxylated porous microspheres according to claim 1, wherein the weight of the oil-soluble emulsifier in step S1 is 1-10% of the weight of the dispersed phase.
7. 7. The method of preparing carboxylated porous microspheres according to claim 1, wherein the weight content of the water soluble emulsifier and/or protective colloid in the continuous phase in step S2 is 0.5-10wt%.
8. 8. The method of preparing carboxylated porous microspheres according to claim 1, wherein the weight ratio of the dispersed phase to the continuous phase in step S3 is 0.1 to 1:1.
9. 9. The method for preparing carboxylated porous microspheres according to claim 1, wherein the continuous stirring in step S3 is performed until precipitation occurs, and gas is continuously introduced from the bottom at the beginning of the continuous stirring.
10. 10. The method of preparing carboxylated porous microspheres according to claim 1, wherein the continuous stirring in step S3 is performed until precipitation occurs, and the carboxyl crosslinking agent is added at the beginning of the continuous stirring.

Description

Preparation method of carboxylated porous microspheres Technical Field The application relates to the technical field of microspheres, in particular to a preparation method of carboxylated porous microspheres. Background Porous microspheres are a raw material commonly used in biological reagent detection, and are particularly used for separating and purifying nucleic acid, so that the porous surface is required to have active functional groups such as hydroxyl groups, carboxyl groups, amino groups and the like, so that the hydrophilicity and the binding property of the porous microspheres with the nucleic acid can be improved, and the separating and purifying effects of the nucleic acid can be improved. The carboxyl functional porous microsphere is usually treated by adopting a carboxylation reagent, such as oleic acid, undecanoic acid and the like, but the physical combination is adopted, the molecular weight of long-chain alkyl carboxylic acid is small, the stability after carboxylation is not high enough or carboxyl groups cannot be effectively present on the surface of the porous microsphere, so that the performance of the carboxyl functional porous microsphere cannot be effectively reflected, and the application effect on nucleic acid extraction and purification is poor. Based on this, there is still a need for improvements in the art of carboxylated porous microspheres. Disclosure of Invention In order to solve the technical problems, the application provides a preparation method of carboxylated porous microspheres, which improves the application effect of the carboxylated porous microspheres on nucleic acid extraction and purification. The application adopts the following technical scheme: the preparation method of the carboxylated porous microspheres comprises the following steps: S1, adding a biocompatible polymer, an acrylic acid copolymer and an oil-soluble emulsifier into an organic solvent for dissolution to prepare a disperse phase; S2, adding a water-soluble emulsifier and/or a protective colloid into water to prepare a continuous phase; And S3, mixing the disperse phase in the step S1 and the continuous phase in the step S2 to obtain emulsion, continuously stirring until precipitation appears, collecting the precipitation, cleaning, and drying to obtain the carboxylated porous microspheres. Preferably, the weight ratio of the biocompatible polymer to the acrylic copolymer in step S1 is 1:0.01-0.2. Preferably, the weight proportion of acrylic acid in the acrylic acid copolymer in step S1 is 1-50%. Preferably, the organic solvent in step S1 has a boiling point of not more than 120 ℃ at 1 atmosphere. Preferably, the concentration of the dispersed phase in step S1 is 1-60wt%. Preferably, the weight of the oil-soluble emulsifier in step S1 is 1-10% of the weight of the dispersed phase. Preferably, the water-soluble emulsifier and/or protective colloid in step S2 is present in an amount of 0.5 to 10wt% in the continuous phase. Preferably, the weight ratio of the dispersed phase to the continuous phase in step S3 is 0.1-1:1. Preferably, in step S3, the stirring is continued until precipitation occurs, and gas is continuously introduced from the bottom at the beginning of the continuous stirring. Preferably, the stirring is continued in step S3 until precipitation occurs, and the carboxyl crosslinking agent is added at the beginning of the continuous stirring. In summary, the application has the following beneficial effects: 1. The application adopts an emulsification method to prepare the microsphere, and in the process of stirring the emulsion, the surface of the microsphere generates a plurality of holes along with the volatilization of the organic solvent in a disperse phase (oil phase), and the microsphere is precipitated and separated out. The acrylic acid copolymer is adopted to provide carboxyl for the porous microsphere, the molecular weight of the acrylic acid copolymer is larger, the combination effect of the acrylic acid copolymer and the biocompatible polymer is higher, and small molecular fatty acid such as oleic acid and the like can not plasticize the biocompatible polymer, so that more stable carboxyl content and distribution can be provided, and the extraction and purification of nucleic acid are more facilitated. 2. In the application, the emulsion is stirred and the organic solvent volatilizes, and the air is continuously blown from the bottom, so that the volatilization of the organic solvent can be quickened, more and bigger micropores are formed, the precipitation of microspheres is quickened, the air blowing can prevent the microspheres from being precipitated to the bottom of a container too early in the forming process, and the microspheres are deformed due to the action of gravity when not completely solidified and shaped, so that the porous microspheres with higher roundness cannot be obtained. When the porous microsphere with high roundness is used for extracting nucleic