CN101507916B - Preparation method of molecularly imprinted polymer microspheres of macrolide antibiotics - Google Patents

Abstract

The invention discloses a method for preparing macrolide antibiotics molecular engram polymer microspheres, which is characterized by comprising the following steps: dissolving a dispersant, namely polyvinyl alcohol or hydroxyethyl cellulose into secondary distilled water to prepare a water-phase dispersion liquid; dissolving engram molecules and functional monomers into an organic solvent to prepare an oil-phase mixture; adding the oil-phase mixture into the water-phase dispersion liquid under the action of stirring, adding an initiator, namely azo-bis-iso-butyrynitrile into the mixture, performing thermal initiation polymerization on the mixture in water bath, and obtaining polymer microspheres; and adopting an ultrasonic extraction method to elute the engram molecules in a butyl acetate aqueous solution or a methanol solution of acetic acid, using distilled water to wash the engram molecules, and performing vacuum drying on the engram molecules to obtain the macrolide antibiotics molecular engram polymer microspheres. Through the method, the macrolide antibiotics molecular engram polymer microspheres are prepared in water phases and are recognized in the water phases; the reorganization result is close to that obtained by a natural biological molecular recognition system; and the invention provides a method for recognizing, separating and analyzing hydrophilic medicaments .

Description

Preparation method of molecularly imprinted polymer microspheres of macrolide antibiotics

technical field

The invention relates to a method for preparing molecularly imprinted polymer microspheres, in particular to a method for preparing macrolide antibiotic molecularly imprinted polymer microspheres. the

Background technique

Macrolide antibiotics are basic medium-spectrum antibiotics that are widely used. In terms of chemical structure, they are all based on 12-16-membered macrolides, linked by glycoside bonds and 1-3 molecules of sugar. polyoxymacrolide antibiotics. Erythromycin (EM) and roxithromycin are the most commonly used 14-membered macrolide antibiotics. Azithromycin is a semi-synthetic C-9 tertiary amine derivative of a 15-membered lactone ring, and midecamycin and acetylspiramycin are semi-synthetic 16-membered macrolide antibiotics. Semi-synthetic midecamycin and spiramycin derivatives. Erythromycin is a biological fermentation antibiotic, and the rest are semi-synthetic macrolide antibiotics. the

In the fermentation system and synthesis system of the above-mentioned antibiotics, there are multiple components with very similar structures, which have different antibacterial activities and toxicity. In order to obtain high-purity antibacterial drugs, the components must be separated during the refining and synthesis process. For example, the structures of erythromycin A and erythromycin C are very similar, but the antibacterial activity of erythromycin C is much lower than that of erythromycin A. However, its toxicity is much higher than that of erythromycin A, and it is an impurity in the drug. As a drug, erythromycin C and erythromycin A must be separated to improve the antibacterial activity of the drug and reduce toxicity. Due to their very similar structures, it is difficult to separate them during extraction and synthesis. the

Molecularly imprinted polymers of macrolide antibiotics prepared by molecular imprinting technology can use their unique steric structure and specific recognition to "memory" the molecular configuration of macrolide antibiotics, so as to have the ability to specific isotropic Ability to recognize and select constructs. the

For many years, the research on the preparation of molecularly imprinted polymers has mainly focused on non-aqueous solvent systems. Water molecules are considered to be strong polar solvents that weaken the hydrogen bond between imprinted molecules and functional monomers, and are easy to damage the relationship between imprinted molecules and functional groups. If the conditions of imprinting are not properly selected, it is likely to lead to the failure of imprinting. Most of the biomolecular recognition systems are carried out in the aqueous environment, and the application effect of molecularly imprinted polymers prepared in non-aqueous systems is poor in the aqueous environment, and imprinting in organic solvents cannot simulate practical applications in most cases. The aqueous environment in the medium cannot meet the diverse requirements of the use environment. the

The document "Patent Application No. 200610023903.7 Patent for Invention" discloses a method for preparing chloramphenicol molecularly imprinted polymer microspheres. Ball, chloramphenicol belongs to phenylalkylamine antibiotics. According to the literature search, there is no relevant report on the preparation of molecularly imprinted polymer microspheres of macrolide antibiotics in aqueous phase by molecular imprinting technology. the

Invention content

In order to overcome the deficiency that molecularly imprinted polymer microspheres of macrolide antibiotics cannot be prepared in aqueous phase by molecular imprinting technology in the prior art, the present invention provides a preparation method of molecularly imprinted polymer microspheres of macrolide antibiotics , to prepare molecularly imprinted polymer microspheres of macrolide antibiotics in aqueous phase. the

The technical solution adopted by the present invention to solve the technical problem: a method for preparing macrolide antibiotic molecularly imprinted polymer microspheres, which is characterized by comprising the following steps:

(a) Dissolve the dispersant polyvinyl alcohol or hydroxyethyl cellulose in double-distilled water at 50-90°C, stir until completely dissolved, and form an aqueous dispersion. The amount of the dispersant is 0.8% to the amount of the functional monomer 1.5%;

(b) Dissolve the imprinted molecules and functional monomers in an organic solvent, and after ultrasonication for 30-60 minutes at 20-30°C, add a cross-linking agent and apply ultrasonication for 10-20 minutes to form an oil-phase mixture; imprinted molecules: functional Monomer: crosslinking agent = 1:2～6:5～40 (molar ratio);

(c) under stirring at 400-500r/min, slowly add the oil phase mixed liquid prepared in step (b) dropwise to the water phase dispersion liquid prepared in step (a), the oil phase mixed liquid and the water phase dispersion liquid The volume ratio is 1:6~14; the dropping rate is controlled at 2~3ml/min; N ₂ is introduced for 15~30min, and the flow rate of N ₂ is 0.5~1ml/min;

(d) under the action of stirring at 400～500r/min, add initiator azobisisobutyronitrile to the mixed solution prepared in step (c), adopt thermal initiation to carry out suspension polymerization reaction, the amount of initiator is the amount of functional monomer 0.5-1.5%, thermally initiate polymerization in a water bath at 50-70°C for 10-24 hours, cool to room temperature, and filter with suction to obtain polymer microspheres;

(e) The polymer microspheres obtained in step (d) are in 20-30°C, 10-15% butyl acetate aqueous solution or 10-15% acetic acid methanol solution, and the imprinted molecules are eluted by ultrasonic extraction, ultrasonically The elution time is 30-60 minutes until no antibiotics can be detected;

(f) wash repeatedly with distilled water, remove residual butyl acetate, methyl alcohol and acetic acid;

(g) vacuum-drying the eluted molecularly imprinted polymer microspheres at 50-60° C. to obtain macrolide antibiotic molecularly imprinted polymer microspheres;

The imprinted molecule is any one of erythromycin, roxithromycin, azithromycin, 9,3"-diacetylmedecamycin or acetylspiramycin; the functional monomer is α-methacrylic acid , any one of acrylic acid or acrylamide; the crosslinking agent is ethylene glycol dimethyl diacrylate, trimethylol propane triacrylate or N', N'-dimethylbisacrylamide glycol Any one of dimethacrylate; the organic solvent is any one of chloroform, acetonitrile, toluene, butyl acetate, acetic acid or methanol.

The beneficial effects of the present invention are: the present invention prepares molecularly imprinted polymer microspheres of macrolide antibiotics in the water phase, which can carry out molecular recognition in the water phase, which is closer to the natural biomolecular recognition system, and is a pro- Aqueous pharmaceuticals provide methods for identification, separation and analysis in the aqueous phase. the

The present invention is described in detail below in conjunction with embodiment. the

Detailed ways

Example 1: Weigh 0.622g of polyvinyl alcohol and add it to 100ml double distilled water, heat up to 80°C with stirring to dissolve it, and form an aqueous dispersion, cool to room temperature and transfer to a 250ml reactor. Weigh 0.734g of erythromycin and dissolve it in 10ml of toluene, add 0.4ml of acrylic acid, and apply ultrasound for 40min at 30°C to make the erythromycin and acrylic acid fully react to form a complex, then add 4.655g of ethylene glycol dimethyl bis Acrylate, ultrasonic 10min, into oil phase mixture. Slowly add 1 part of the oil phase mixed liquid into 6 parts of the aqueous phase dispersion under stirring at 450r/min, and the dropping speed is controlled at 2ml/min to form a milky white microsuspension emulsion. Introduce N ₂ for 15 minutes, and the flow rate of N ₂ is 0.5ml/min. Then 0.0760 g of azobisisobutyronitrile was added and stirred at 400 r/min, and thermally initiated polymerization in a water bath at 50° C. for 24 hours. After cooling to room temperature, it was filtered to obtain polymer microspheres.

The above-mentioned polymer microspheres were eluted with ultrasonic extraction in 10% butyl acetate aqueous solution at 20°C, and the ultrasonic elution time was 30 minutes until no erythromycin molecules could be detected at 492nm. . Then wash repeatedly with distilled water to remove residual methanol and acetic acid solution. The obtained polymer microspheres were vacuum-dried at 60° C. to obtain erythromycin molecularly imprinted polymer microspheres. the

After testing, the prepared erythromycin molecularly imprinted polymer microspheres had an average particle size of 60 μm. The competing substrate was roxithromycin with a separation factor of 1.87. the

Example 2: Weigh 0.0451 g of hydroxyethyl cellulose and add it to 90 ml of double-distilled water, heat up to 50° C. to dissolve it under stirring, and form an aqueous dispersion. After cooling to room temperature, transfer it to a 250 ml reactor. Weigh 0.734g of erythromycin and dissolve it in 7.5ml of acetonitrile, add 0.4ml of α-methacrylic acid, and apply ultrasound at 25°C for 30min to fully react the erythromycin and methacrylic acid to form a complex, then add 3.960g Ethylene glycol dimethyl diacrylate, ultrasonic 20min, to form an oil phase mixture. Slowly add 1 part of the oil phase mixed liquid into 14 parts of the aqueous phase dispersion under stirring at 400r/min, and the dropping speed is controlled at 3ml/min to form a milky white microsuspension emulsion. Introduce N ₂ for 30 minutes, and the flow rate of N ₂ is 1ml/min. Then 0.0760 g of azobisisobutyronitrile was added and stirred, the stirring condition was 500 r/min, and the polymerization was thermally initiated in a 70° C. water bath for 20 hours. After cooling to room temperature, it was filtered to obtain polymer microspheres.

The above polymer microspheres were eluted with ultrasonic extraction in 15% butyl acetate aqueous solution at 30°C, and the ultrasonic elution time was 60 minutes until no erythromycin molecules were detected at 492nm. , and then repeatedly washed with distilled water to remove residual methanol and acetic acid solution. The obtained polymer microspheres were vacuum dried at 50°C. The erythromycin molecularly imprinted polymer microspheres were obtained. the

After testing, the average particle size of the prepared erythromycin molecularly imprinted polymer microspheres was 45 μm. The competing substrate was roxithromycin with a separation factor of 2.25. the

Example 3: Weigh 0.055 g of hydroxyethyl cellulose and add it to 10 ml of double-distilled water, heat up to 80° C. to dissolve it under stirring, and form an aqueous dispersion, and transfer it to a 250 ml reactor after cooling to room temperature. Weigh 0.734g of erythromycin and dissolve it in 7.5ml of acetonitrile, add 0.4ml of α-methacrylic acid, and apply ultrasound for 50min at 20°C to fully react the erythromycin and α-methacrylic acid to form a complex, then add 3.960 g ethylene glycol dimethyl diacrylate, ultrasonic 10min, an oil phase mixture. Slowly add 1 part of the oil-phase mixed liquid into 7 parts of the aqueous phase dispersion under stirring at 500 r/min, and the dropping speed is controlled at 2 ml/min to form a milky white micro-suspoemulsion. Introduce N ₂ for 20 minutes, and the flow rate of N ₂ is 0.6ml/min. Then 0.0760 g of azobisisobutyronitrile was added and stirred at 450 r/min, and the polymerization was thermally initiated in a water bath at 65° C. for 20 hours. After cooling to room temperature, it was filtered to obtain polymer microspheres.

The above-mentioned polymer microspheres were eluted with ultrasonic extraction in 13% butyl acetate aqueous solution at 25°C, and the ultrasonic elution time was 40 min until no erythromycin molecules were detected at 492 nm. , and then repeatedly washed with distilled water to remove residual methanol and acetic acid solution. The obtained polymer microspheres were vacuum dried at 55°C. The erythromycin molecularly imprinted polymer microspheres were obtained. the

After testing, the average particle size of the prepared erythromycin molecularly imprinted polymer microspheres was 40 μm. The competing substrate was roxithromycin with a separation factor of 2.18. the

Example 4: Weigh 0.622g of polyvinyl alcohol and add it to 120ml of twice-distilled water, heat it up to 90°C with stirring to dissolve it, and form an aqueous dispersion. After cooling to room temperature, transfer it into a 250ml reactor. Weigh 0.825g of roxithromycin and dissolve it in 9ml of chloroform, add 0.5ml of acrylic acid, and apply ultrasound for 40 minutes at 20°C to make the roxithromycin and acrylic acid fully react to form a complex, then add 2.90g of trimethylolpropane Triacrylate, ultrasonic 15min, to form an oil phase mixture. Slowly add 1 part of the oil phase mixed solution dropwise to 8 parts of the aqueous phase dispersion under stirring at 410 r/min, and the dropping rate is controlled at 3 ml/min to form a milky white microsuspension emulsion. Introduce N ₂ for 18 minutes, and the flow rate of N ₂ is 0.7ml/min. Then 0.0760 g of azobisisobutyronitrile was added and stirred, the stirring condition was 410 r/min, and the polymerization was thermally initiated in a 65° C. water bath for 18 hours. After cooling to room temperature, it was filtered to obtain polymer microspheres.

The above-mentioned polymer microspheres were eluted with ultrasonic extraction in 11% butyl acetate aqueous solution at 22°C, and the ultrasonic elution time was 50 min until no erythromycin molecules were detected at 492 nm. , and then repeatedly washed with distilled water to remove residual methanol and acetic acid solution, and the obtained polymer microspheres were vacuum-dried at 54°C. Roxithromycin molecularly imprinted polymer microspheres were obtained. the

After testing, the prepared roxithromycin molecularly imprinted polymer microspheres had an average particle size of 50 μm. The competing substrate was azithromycin with a separation factor of 1.62. the

Example 5: Weigh 0.0521 g of hydroxyethyl cellulose and add it to 100 ml of twice-distilled water, heat up to 60° C. to dissolve it under stirring, and form an aqueous dispersion. After cooling to room temperature, transfer it into a 250 ml reactor. Weigh 0.825g of roxithromycin and dissolve it in 9ml of chloroform, add 0.5ml of acrylic acid, and apply ultrasound at 30°C for 40 minutes to make the roxithromycin and acrylic acid fully react to form a complex, then add 2.90g of trimethylolpropane Triacrylate, ultrasonic 15min, to form an oil phase mixture. 1 part of the oil phase mixed solution was slowly added dropwise to 9 parts of the aqueous phase dispersion under stirring at 420r/min, and the rate of addition was controlled at 2ml/min to form a milky white microsuspension emulsion. Introduce N ₂ for 16 minutes, and the flow rate of N ₂ is 0.8ml/min. Then 0.0760 g of azobisisobutyronitrile was added and stirred at 430 r/min, and thermally initiated polymerization in a water bath at 55° C. for 18 hours. After cooling to room temperature, it was filtered to obtain polymer microspheres.

The above polymer microspheres were eluted with ultrasonic extraction in 14% butyl acetate aqueous solution at 26°C, and the ultrasonic elution time was 35 minutes until no erythromycin molecules could be detected at 492nm. , and then repeatedly washed with distilled water to remove residual methanol and acetic acid solutions, and the obtained polymer microspheres were vacuum-dried at 53° C. to obtain roxithromycin molecularly imprinted polymer microspheres. the

After testing, the average particle size of the prepared roxithromycin molecularly imprinted polymer microspheres was 40 μm. The competing substrate was azithromycin with a separation factor of 1.93. the

Example 6: Weigh 0.622g of polyvinyl alcohol and add it to 120ml of double distilled water, stir and heat up to 80°C to dissolve it, forming an aqueous dispersion, and transfer it to a 250ml reactor after cooling to room temperature. Weigh 0.880g of azithromycin and dissolve it in 7.5ml of acetonitrile, add 0.3g of acrylamide, and apply ultrasound at 25°C for 60min to fully react the azithromycin and acrylamide to form a complex, then add 4.655g of ethylene di Alcohol dimethyl diacrylate, ultrasonic 10min, to form an oil phase mixture. Slowly add 1 part of the oil-phase mixed liquid into 10 parts of the aqueous phase dispersion under stirring at 430 r/min, and the dropping speed is controlled at 3 ml/min to form a milky white microsuspension emulsion. N ₂ was introduced for 17 minutes, and the flow rate of N ₂ was 0.9ml/min. Then 0.0760g of azobisisobutyronitrile was added and stirred under the stirring condition of 440r/min, thermally initiated polymerization in a 60°C water bath for 16 hours, cooled to room temperature, and filtered to obtain polymer microspheres.

The above polymer microspheres were eluted with ultrasonic extraction in 12% butyl acetate aqueous solution at 28°C, and the ultrasonic elution time was 45 minutes until no erythromycin molecules could be detected at 492nm. , and then repeatedly washed with distilled water to remove residual methanol and acetic acid solution. The obtained polymer microspheres were vacuum dried at 52°C. Azithromycin molecularly imprinted polymer microspheres were obtained. the

After testing, the prepared azithromycin molecularly imprinted polymer microspheres had an average particle size of 60 μm. The competing substrate was roxithromycin with a separation factor of 2.35. the

Example 7: Weigh 0.0451g of hydroxyethyl cellulose and add it into 90ml of twice-distilled water, stir and heat up to 60°C to dissolve it to form an aqueous dispersion, cool to room temperature and transfer it into a 250ml reactor. Weigh 0.880g of azithromycin and dissolve it in 7.5ml of acetonitrile, add 0.3g of acrylamide, and apply ultrasound for 60min at 20°C to make the azithromycin and acrylamide fully react to form a complex, then add 4.655g of acetonitrile Glycol dimethyl diacrylate, ultrasonic 10min, into an oil phase mixture. Slowly add 1 part of the oil phase mixed liquid into 11 parts of the aqueous phase dispersion under stirring at 460r/min, and the dropping speed is controlled at 2ml/min to form a milky white microsuspension emulsion. Introduce N ₂ for 22 minutes, and the flow rate of N ₂ is 0.5ml/min. Then, 0.0760 g of azobisisobutyronitrile was added and stirred. The stirring condition was 460 r/min, and thermally initiated polymerization in a 65°C water bath for 16 hours. After cooling to room temperature, the mixture was filtered to obtain polymer microspheres.

The above-mentioned polymer microspheres were eluted with ultrasonic extraction in methanol solution of 10% acetic acid at 20° C., and the ultrasonic elution time was 55 min until no erythromycin molecules were detected at 492 nm. Then wash repeatedly with distilled water to remove residual methanol and acetic acid solution. The obtained polymer microspheres were vacuum dried at 51°C. Azithromycin molecularly imprinted polymer microspheres were obtained. the

After testing, the prepared azithromycin molecularly imprinted polymer microspheres had an average particle size of 45 μm. The competing substrate was roxithromycin with a separation factor of 2.18. the

Example 8: Weigh 0.0451 g of hydroxyethyl cellulose and add it to 90 ml of double-distilled water, stir and heat up to 50°C to dissolve it, forming an aqueous dispersion, and transfer it to a 250 ml reactor after cooling to room temperature. Weigh 0.850g of 9,3”-diacetyl midecamycin and dissolve it in 8ml of acetonitrile, add 0.4ml of acrylic acid, and apply ultrasound at 20°C for 40min to fully react with 9,3”-diacetyl midecamycin and acrylic acid to form To the compound, add 2.16g of N',N'-2 methylbisacrylamide glycol dimethacrylate, and sonicate for 20 minutes to form an oil phase mixture. Slowly add 1 part of the oil phase mixed liquid into 12 parts of the aqueous phase dispersion under stirring at 470r/min, and the dropping speed is controlled at 2ml/min to form a milky white microsuspension emulsion. Introduce N ₂ for 24 minutes, and the flow rate of N ₂ is 0.6ml/min. Then 0.0469g of azobisisobutyronitrile was added and stirred at 470r/min, and thermally initiated polymerization in a water bath at 55°C for 20 hours. After cooling to room temperature, it was filtered to obtain polymer microspheres.

The above-mentioned polymer microspheres were eluted by ultrasonic extraction in methanol solution of 15% acetic acid at 30°C, and the ultrasonic elution time was 30 minutes until no erythromycin molecules were detected at 492nm. Then wash repeatedly with distilled water to remove residual methanol and acetic acid solution. The obtained polymer microspheres were vacuum dried at 56°C. 9,3"-diacetyl midecamycin molecularly imprinted polymer microspheres were obtained. 

After testing, the prepared 9,3"-diacetyl midecamycin molecularly imprinted polymer microspheres had an average particle size of 55 μm. The competitive substrate was erythromycin, and the separation factor was 1.88. 

Example 9: Weigh 0.622g of polyvinyl alcohol and add it into 120ml of double distilled water, stir and heat up to 90°C to dissolve it, forming an aqueous dispersion, and transfer it to a 250ml reactor after cooling to room temperature. Weigh 0.850g of 9,3”-diacetyl midecamycin and dissolve it in 8ml of acetonitrile, add 0.3g of acrylamide, and apply ultrasound at 25°C for 60min to make 9,3”-diacetyl midecamycin and acrylamide fully After forming a complex, add 2.16g of N', N'-2 methylbisacrylamide ethylene glycol dimethacrylate, and sonicate for 20 minutes to form an oil phase mixture. Slowly add 1 part of the oil phase mixed liquid into 13 parts of the aqueous phase dispersion under stirring at 480r/min, and the dropping speed is controlled at 3ml/min to form a milky white microsuspension emulsion. Introduce N ₂ for 26 minutes, and the flow rate of N ₂ is 0.8ml/min. Then 0.0469g of azobisisobutyronitrile was added and stirred at 480r/min, and thermally initiated polymerization in a water bath at 60°C for 20 hours. After cooling to room temperature, it was filtered to obtain polymer microspheres.

The above-mentioned polymer microspheres were eluted with ultrasonic extraction in methanol solution of 13% acetic acid at 25°C, and the ultrasonic elution time was 60 min until no erythromycin molecules were detected at 492 nm. Then wash repeatedly with distilled water to remove residual methanol and acetic acid solution. The obtained polymer microspheres were vacuum dried at 57°C. 9,3"-diacetyl midecamycin molecularly imprinted polymer microspheres were obtained. 

After testing, the prepared 9,3"-diacetyl midecamycin molecularly imprinted polymer microspheres had an average particle size of 60 μm. The competitive substrate was erythromycin, and the separation factor was 1.91. 

Example 10: Weigh 0.0451 g of hydroxyethyl cellulose and add it to 90 ml of double-distilled water, stir and heat up to 50°C to dissolve it, forming an aqueous dispersion, and transfer it to a 250 ml reactor after cooling to room temperature. Weigh 0.922g of acetylspiramycin and dissolve it in 8ml of acetonitrile, add 0.3g of acrylamide, and apply ultrasound at 30°C for 40 minutes to make the acetylspiramycin and acrylamide fully react to form a complex, then add 2.16g of N', N'-2 methylbisacrylamide ethylene glycol dimethacrylate, sonicated for 10 minutes to form an oil phase mixture. Slowly add 1 part of the oil phase mixed solution into 10 parts of the aqueous phase dispersion under stirring at 490 r/min, and the dropping rate is controlled at 2 ml/min to form a milky white microsuspension emulsion. Introduce N ₂ for 28 minutes, and the flow rate of N ₂ is 0.5ml/min. Then 0.0469g of azobisisobutyronitrile was added and stirred, the stirring condition was 490r/min, and the polymerization was thermally initiated in a 50°C water bath for 16 hours. After cooling to room temperature, it was filtered to obtain polymer microspheres.

The above-mentioned polymer microspheres were eluted with ultrasonic extraction in methanol solution of 11% acetic acid at 26°C, and the ultrasonic elution time was 40 min until no erythromycin molecules were detected at 492 nm. Then wash repeatedly with distilled water to remove residual methanol and acetic acid solution. The obtained polymer microspheres were vacuum dried at 58°C. Acetylspiramycin molecularly imprinted polymer microspheres were obtained. the

After testing, the average particle size of the prepared acetylspiramycin molecularly imprinted polymer microspheres was 47 μm. The competing substrate was midecamycin with a separation factor of 1.93. the

Example 11: Weigh 0.622g of polyvinyl alcohol and add it to 90ml of double distilled water, stir and heat up to 90°C to dissolve it, forming an aqueous dispersion, and transfer it to a 250ml reactor after cooling to room temperature. Weigh 0.922g of acetylspiramycin and dissolve it in 8ml of acetonitrile, add 0.5ml of methacrylic acid, and apply ultrasound at 30°C for 60 minutes to fully react the acetylspiramycin and methacrylic acid to form a complex, then add 2.16g of N' , N'-2 methyl bisacrylamide ethylene glycol dimethacrylate, ultrasonic 20min, an oil phase mixture. Slowly add 1 part of the oil-phase mixed liquid to 8 parts of the aqueous phase dispersion under stirring at 450 r/min, and the dropping speed is controlled at 3 ml/min to form a milky white micro-suspoemulsion. N ₂ was introduced for 29 minutes, and the flow rate of N ₂ was 0.9ml/min. Then 0.0469g of azobisisobutyronitrile was added and stirred, the stirring condition was 410r/min, and the polymerization was thermally initiated in a 70°C water bath for 18 hours. After cooling to room temperature, it was filtered to obtain polymer microspheres.

The above-mentioned polymer microspheres were eluted with ultrasonic extraction in 12% methanol solution of acetic acid at 21°C, and the imprinted molecules were eluted for 50 minutes until no erythromycin molecules were detected at 492 nm. Then wash repeatedly with distilled water to remove residual methanol and acetic acid solution. The obtained polymer microspheres were vacuum dried at 59°C. Acetylspiramycin molecularly imprinted polymer microspheres were obtained. the

After testing, the prepared acetylspiramycin molecularly imprinted polymer microspheres had an average particle size of 55 μm. The competing substrate was midecamycin with a separation factor of 1.95. the

Claims (1)

1. the preparation method of a macrolide antibiotics molecular engram polymer microsphere is characterized in that may further comprise the steps:

(a) polyethylene of dispersing agent alcohol or hydroxyethylcellulose are dissolved in 50～90 ℃ the redistilled water, are stirred to whole dissolvings, become aqueous dispersion liquid, dispersant dosage is 0.8%～1.5% of a function monomer consumption;

(b) microsphere and function monomer are dissolved in the organic solvent, behind ultrasonication 30～60min, add crosslinking agent under 20～30 ℃ of conditions, ultrasonication 10～20min becomes the oil phase mixed liquor; Microsphere: function monomer: crosslinking agent=1: 2～6: 5～40 (mol ratio);

(c) under 400～500r/min stirring action, the oil phase mixed liquor that step (b) is prepared slowly is added drop-wise in the aqueous dispersion liquid of step (a) preparation, and the volume ratio of oil phase mixed liquor and aqueous dispersion liquid is 1: 6～14; Rate of addition is controlled at 2～3ml/min; Feed N ₂15～30min, N ₂Flow 0.5～1ml/min;

(d) under 400～500r/min stirring action, in the mixed liquor of step (c) preparation, add the initator azodiisobutyronitrile, adopt heat to cause and carry out suspension polymerization, initiator amount is 0.5～1.5% of a function monomer consumption, thermal-initiated polymerization 10～24h in 50～70 ℃ of water-baths, be cooled to room temperature, suction filtration is handled, and obtains polymer microballoon;

(e) polymer microballoon that step (d) is obtained is in the methanol solution of 20～30 ℃, 10～15% butyl acetate aqueous solution or 10～15% acetate, adopt the method wash-out microsphere of ultrasonic extraction, ultrasonic elution time is 30～60min, till can not detecting antibiotic;

(f) use the distilled water cyclic washing, remove residual butyl acetate, methyl alcohol and acetate;

(g) with the molecular blotting polymer microsphere behind the wash-out 50～60 ℃ of following vacuum drying, obtain macrolide antibiotics molecular engram polymer microsphere;

Described microsphere is erythromycin, ROX, azithromycin, 9,3 "-any of diacetyl medecamycin or acetyl spiramycin; Described function monomer is any of α-Jia Jibingxisuan, acrylic acid or acrylamide; Described crosslinking agent is trimethylolpropane triacrylate, N ', any of N '-dimethyl bisacrylamide or ethylene glycol dimethacrylate; Described organic solvent is any of chloroform, acetonitrile, toluene, butyl acetate, acetate or methyl alcohol.