CN104792688A - Acid-soluble chitosan microsphere and preparation method thereof - Google Patents
Acid-soluble chitosan microsphere and preparation method thereof Download PDFInfo
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- CN104792688A CN104792688A CN201510142199.6A CN201510142199A CN104792688A CN 104792688 A CN104792688 A CN 104792688A CN 201510142199 A CN201510142199 A CN 201510142199A CN 104792688 A CN104792688 A CN 104792688A
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Abstract
The invention provides acid-soluble chitosan microspheres which are prepared from chitosan and terephthalaldehyde through cross-linking reaction. The particle size of the microspheres is 6-11mu m, and the variable coefficient of the microspheres is 2.7-5%. The preparation method comprises the following steps: (1) preparing ejection liquid and receiving liquid; (2) in a constant temperature environment of 25-45 DEG C, adding the ejection liquid of the temperature identical to that of the constant temperature environment into an injector of electrostatic spinning equipment, pushing the ejection liquid into a metal syringe needle through an ejection pump, exerting high-voltage static electricity to the metal syringe needle so as to form the ejection liquid into ejection liquid droplets, receiving the ejection liquid droplets by using a container with the receiving liquid under the stirring condition, and enabling chitosan in the ejection liquid droplets to be completely cross-linked with terephthalaldehyde in the receiving liquid, thereby obtaining the chitosan microspheres; (3) washing the chitosan microspheres by using isopropanol so as to remove the receiving liquid on the surfaces of the microspheres, and washing with water so as to remove isopropanol.
Description
Technical field
The invention belongs to chitosan microball field, particularly a kind of single dispersing solubility in acid chitosan microball with erythrocyte size and preparation method thereof.
Background technology
Along with the widespread use in medical laboratory instrument such as electronic technology, Flow Cytometry, laser technology and robot calculator, the automaticity of cellanalyzer (flow cytometer) improve constantly and be widely used in clinical in.The data measured due to cellanalyzer are relative value, therefore must calibrate instrument system or demarcate before use.At present, standard items for calibrating cellanalyzer are behaved or the erythrocyte (see U.S.Pat.No.4489162) of chicken, but from mammalian, extract erythrocyte there is preparation process complexity, there is very high bio-hazard and the problem such as with high costs, and the shelf-life of this kind of " pure natural " erythrocyte is very short.For this reason, people wish to prepare the standard items that polymer microballoon is calibrated to replace erythrocyte as cellanalyzer.Zelmanovic etc. disclose the polymer microballoon that erythrocyte can be replaced as standard items, and confirm that namely the polymer microballoon meeting the condition such as good stability in particle diameter and erythrocyte sizableness, uniform particle diameter, nontoxic, calibration testing process can be used as cellanalyzer calibration criterion product and use (see U.S.Pat.No.6074879) by experiment.
Due to shitosan there is unique biologically active, nontoxic, biocompatibility good and be easy to the features such as degraded, usually chitosan microball is prepared or micro-capsule uses as pharmaceutical carrier, but there is no at present and shitosan is prepared into microballoon and is used for the report that replacement blood red blood cell uses as the standard items that blood cell analysis instrument is calibrated, this is mainly because existing method is difficult to shitosan to be prepared into particle diameter and erythrocyte sizableness and the good microballoon of monodispersity.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, a kind of solubility in acid chitosan microball and preparation method thereof is provided, to improve the monodispersity of the particle diameter chitosan microball suitable with erythrocyte, and enrich the kind of blood cell analysis instrument calibration standard product.
Solubility in acid chitosan microball of the present invention, is formed by shitosan and terephthalaldehyde cross-linking reaction, and the particle diameter of microballoon is 6 ~ 11 μm, and the value for coefficient of variation of microballoon is 2.7% ~ 5%.
The principle that chitosan microball of the present invention dissolves in acid condition is such as formula shown in (1), in neutral medium, the amino of shitosan and the aldehyde radical of terephthalaldehyde react and generate schiff bases, therefore in neutral conditions, chitosan microball can keep the integrality of good sphericity and structure; In the acid medium of low ph value, shitosan is positively charged due to protonated amino, intermolecular electrostatic repulsion and water wettability strengthen, chitosan microball is sharply swelling, constantly protonated along with amino of chitosan, the schiff bases that shitosan and terephthalaldehyde cross-linking reaction are formed becomes unstable gradually, finally causes chitosan microball disintegrate and dissolve.
Present invention also offers a kind of preparation method of above-mentioned solubility in acid chitosan microball, processing step is as follows:
(1) preparation of jetting fluid and receiving liquid
The preparation of jetting fluid: be dissolved in water by water soluble chitosan and form chitosan solution, then regulates pH value to 6.3 ~ 6.5 of chitosan solution with sodium hydrate aqueous solution, by centrifugal for the chitosan solution after adjust ph, filtration, namely form jetting fluid; The mass ratio of described water soluble chitosan and water is (0.02 ~ 0.03): 1;
The preparation of receiving liquid: poly-ricinoleic acid glyceride, tetramethylethylenediamine are mixed with solvent, then add terephthalaldehyde, is stirred to terephthalaldehyde and dissolves namely form receiving liquid completely; Described solvent is the mixed liquor of toluene and n-hexyl alcohol, the volume ratio of toluene and n-hexyl alcohol is (1 ~ 2): 1, the amount of poly-ricinoleic acid glyceride is every 1mL solvent 0.04 ~ 0.08g, the volume ratio of tetramethylethylenediamine and solvent is (0.004 ~ 0.008): 1, and the amount of terephthalaldehyde is every 1mL solvent 0.01 ~ 0.02g;
(2) preparation of chitosan microball
In the isoperibol of 25 ~ 45 DEG C, the jetting fluid identical with isoperibol temperature is added in the syringe of electrospinning device, then metal needle is pushed by syringe pump, high-pressure electrostatic is applied at metal needle place, namely jetting fluid forms liquid droplets, adopt the container filling receiving liquid to receive described liquid droplets under agitation, the shitosan in drop to be sprayed and the terephthalaldehyde in receiving liquid are completely crosslinked, obtain chitosan microball; The voltage controlling to be applied to metal needle place is 5 ~ 6kV, and jet length is 5 ~ 10cm, and the internal diameter of metal needle is 104 ~ 130 μm, and the flow of jetting fluid is 110 ~ 140 μ L/h;
(3) wash
Adopt washed with isopropyl alcohol step (2) gained chitosan microball to remove the receiving liquid of microsphere surface, then wash removing isopropyl alcohol with water.
In the step (2) of said method, to receiving liquid stir be liquid droplets in order to avoid first ejecting receiving liquid surface with after the liquid droplets that ejects merge, and inter-adhesive between the microballoon avoiding occurring in receiving liquid cross-linking reaction, preferentially adopt the rotating speed of 200 ~ 300rpm to stir receiving liquid.
In said method, weight-average molecular weight≤5000 of described water soluble chitosan.
In said method, described water is deionized water or distilled water.
In said method, the concentration of described sodium hydrate aqueous solution is at least 1mol/L.
Compared with prior art, the present invention has following beneficial effect:
1, the invention provides a kind of novel chitosan microball, the particle diameter of this microballoon is 6 ~ 11 μm, with the sizableness of erythrocyte, and uniform particle diameter, there is good sphericity, the monodispersity of this microballoon is very good, the value for coefficient of variation of microballoon is 2.7% ~ 5%, in addition the shitosan that terephthalaldehyde is crosslinked is nontoxic, above-mentioned speciality all meets the requirement of blood cell analysis instrument calibration standard product, therefore chitosan microball of the present invention can be used for replacement blood red blood cell and uses as blood cell analysis instrument calibration standard product, enrich the kind of blood cell analysis instrument calibration standard product.
2, because chitosan microball of the present invention is formed by cross-linking reaction by shitosan and terephthalaldehyde, and the cross-linking reaction between shitosan and terephthalaldehyde is reversible, this chitosan microball can keep good sphericity and structural intergrity in neutral environment, and can dissolve rapidly in sour environment, therefore, after using chitosan microball of the present invention to use as blood cell analysis instrument calibration standard product, the chitosan microball remained in wherein can be dissolved by adding acid in instrument, effectively can avoid microballoon remaining in analytical instrument, thus the accuracy improved when analytical instrument is reused, in addition, the characteristic being cross-linked at alkaline environment according to described chitosan microball, being cross-linked in sour environment solution, pharmaceutical carrier can be it can be used as to use, and at stomach low ph environment targeting drug release, the rapid, effective treatment for disease of stomach provides new pharmaceutical carrier.
3, because the method for the invention adopts low-molecular-weight water soluble chitosan to prepare jetting fluid, and the viscosity etc. of jetting fluid ties in the operating parameter of electrostatic spraying, and therefore the method for the invention has obtained ten micron-sized chitosan microballs; Because the tetramethylethylenediamine in receiving liquid can accelerate the crosslinking rate of shitosan and terephthalaldehyde, and receiving liquid is for solvent is formulated with density and all less toluene of viscosity and n-hexyl alcohol, thus liquid droplets effectively can be avoided to stick together on the surface of receiving liquid and inside merging, thus ensure that chitosan microball prepared by the method for the invention has the monodispersity of good sphericity and height.
4, because the method for the invention adopts electrostatic spraying legal system for chitosan microball, electrostatic spraying method has simple to operate, that equipment is ripe advantage, and cost is also comparatively cheap, therefore the method for the invention is easy to the industrial-scale production realizing chitosan microball product.
Figure of description
Fig. 1 is that the method for the invention adopts electrospinning device to prepare the schematic diagram of chitosan microball, in figure, wherein, and 1-syringe pump, 2-syringe, 3-high-voltage power supply, 4-receiving vessel, 5-magnetic stirring apparatus.
Fig. 2 is the scanning electron microscopic picture of chitosan microball prepared by embodiment 1;
Fig. 3 is the fluorescence picture of chitosan microball prepared by embodiment 1;
Fig. 4 is the fluorescence picture of chitosan microball prepared by embodiment 2;
Fig. 5 is the fluorescence picture of chitosan microball prepared by embodiment 3;
Fig. 6 is chitosan microball prepared by embodiment 10 is the Laser Scanning Confocal Microscope photo of course of dissolution in the damping fluid of 3.5 in pH value.
Embodiment
Below by embodiment, solubility in acid chitosan microball of the present invention and preparation method thereof is described further.
In following each embodiment, molecular weight≤5000 of described water soluble chitosan, purchased from sea, Jinan marine biotechnology company limited of Derby, toluene, n-hexyl alcohol and isopropyl alcohol are purchased from Chengdu Ke Long chemical reagent factory, terephthalaldehyde is purchased from Chemical Reagent Co., Ltd., Sinopharm Group, and poly-ricinoleic acid glyceride (PGPR 90) is purchased from Danisco company of Denmark.
Embodiment 1
In the present embodiment, the preparation method of chitosan microball is as follows:
(1) preparation of jetting fluid and receiving liquid
The preparation of jetting fluid: water soluble chitosan is dissolved and forms chitosan solution in deionized water, then regulate between pH value to 6.3 ~ 6.4 of chitosan solution with the sodium hydrate aqueous solution that concentration is 1mol/L, by centrifugal for the chitosan solution after adjust ph, use Filter paper filtering, namely form jetting fluid; The mass ratio of described water soluble chitosan and deionized water is 0.02:1;
The preparation of receiving liquid: poly-ricinoleic acid glyceride, tetramethylethylenediamine are mixed with solvent, then add terephthalaldehyde, is stirred to terephthalaldehyde and dissolves namely form receiving liquid completely; Described solvent is the mixed liquor of toluene and n-hexyl alcohol, the volume ratio of toluene and n-hexyl alcohol is 1:1, the amount of poly-ricinoleic acid glyceride is every 1mL solvent 0.06g, and the volume ratio of tetramethylethylenediamine and solvent is 0.006:1, and the amount of terephthalaldehyde is every 1mL solvent 0.02g.
(2) preparation of chitosan microball
The temperature regulating electrostatic spinning machine is 45 DEG C, open recirculating air button simultaneously, make the temperature in electrostatic spinning machine and remain on 45 DEG C, the container being loaded with jetting fluid is placed in the space of electrostatic spinning machine, when the temperature of liquid to be sprayed reaches 45 DEG C, jetting fluid to be loaded in the syringe 2 of electrostatic spinning machine and to push metal needle by syringe pump 1, the voltage of 5KV ~ 6KV is applied at metal needle place, namely jetting fluid forms liquid droplets, adopt fill receiving liquid container reception described in liquid droplets adopt magnetic stirring apparatus to stir receiving liquid with the rotating speed of 200rpm, shitosan after 8h in liquid droplets and the terephthalaldehyde in receiving liquid are completely crosslinked, obtain chitosan microball, the flow of described jetting fluid is 110 μ l/h, and the internal diameter of metal needle is 104 μm (metal needle model is 32G), and jet length is 5cm.
(3) wash
Adopt washed with isopropyl alcohol step (2) gained chitosan microball, the receiving liquid of removing microsphere surface, then spend deionized water removing isopropyl alcohol.Fig. 2 is the scanning electron microscopic picture of microballoon prepared by the present embodiment, Fig. 3 is the fluorescence picture of microballoon prepared by the present embodiment, from Fig. 2 and Fig. 3, the uniform particle diameter of microballoon prepared by the present embodiment, sphericity are good, the coefficient of variation (CV) value of chitosan microball is 2.7%, and the particle diameter of chitosan microball is 6.4 μm.
Embodiment 2
In the present embodiment, the preparation method of chitosan microball is as follows:
(1) preparation of jetting fluid and receiving liquid
The preparation of jetting fluid: water soluble chitosan is dissolved and forms chitosan solution in deionized water, then regulate between pH value to 6.4 ~ 6.5 of chitosan solution with the sodium hydrate aqueous solution that concentration is 2mol/L, by centrifugal for the chitosan solution after adjust ph, use Filter paper filtering, namely form jetting fluid; The mass ratio of described water soluble chitosan and deionized water is 0.03:1;
The preparation of receiving liquid: poly-ricinoleic acid glyceride, tetramethylethylenediamine are mixed with solvent, then add terephthalaldehyde, is stirred to terephthalaldehyde and dissolves namely form receiving liquid completely; Described solvent is the mixed liquor of toluene and n-hexyl alcohol, the volume ratio of toluene and n-hexyl alcohol is 1:1, the amount of poly-ricinoleic acid glyceride is every 1mL solvent 0.06g, and the volume ratio of tetramethylethylenediamine and solvent is 0.004:1, and the amount of terephthalaldehyde is every 1mL solvent 0.02g.
(2) preparation of chitosan microball
The temperature regulating electrostatic spinning machine is 45 DEG C, open recirculating air button simultaneously, make the temperature in electrostatic spinning machine and remain on 45 DEG C, the container being loaded with jetting fluid is placed in the space of electrostatic spinning machine, when the temperature of liquid to be sprayed reaches 45 DEG C, jetting fluid to be loaded in the syringe 2 of electrostatic spinning machine and to push metal needle by syringe pump 1, the voltage of 5KV ~ 6KV is applied at metal needle place, namely jetting fluid forms liquid droplets, adopt fill receiving liquid container reception described in liquid droplets adopt magnetic stirring apparatus to stir receiving liquid with the rotating speed of 300rpm, shitosan after 8h in liquid droplets and the terephthalaldehyde in receiving liquid are completely crosslinked, obtain chitosan microball, the flow of described jetting fluid is 110 μ l/h, and the internal diameter of metal needle is 104 μm (metal needle model is 32G), and jet length is 5cm.
(3) wash
Adopt the receiving liquid of washed with isopropyl alcohol step (2) gained chitosan microball removing microsphere surface, then spend deionized water removing isopropyl alcohol.Fig. 4 is the fluorescence picture of microballoon prepared by the present embodiment, and as shown in Figure 4, uniform particle diameter, the sphericity of microballoon prepared by the present embodiment are good, and the coefficient of variation (CV) value of chitosan microball is 5%, and the particle diameter of chitosan microball is 9 μm.
Embodiment 3
In the present embodiment, the preparation method of chitosan microball is as follows:
(1) preparation of jetting fluid and receiving liquid
The preparation of jetting fluid: water soluble chitosan is dissolved in distilled water and forms chitosan solution, then regulate between pH value to 6.4 ~ 6.5 of chitosan solution with the sodium hydrate aqueous solution that concentration is 1mol/L, by centrifugal for the chitosan solution after adjust ph, use Filter paper filtering, namely form jetting fluid; The mass ratio of described water soluble chitosan and distilled water is 0.02:1;
The preparation of receiving liquid: poly-ricinoleic acid glyceride, tetramethylethylenediamine are mixed with solvent, then add terephthalaldehyde, is stirred to terephthalaldehyde and dissolves namely form receiving liquid completely; Described solvent is the mixed liquor of toluene and n-hexyl alcohol, the volume ratio of toluene and n-hexyl alcohol is 1:1, the amount of poly-ricinoleic acid glyceride is every 1mL solvent 0.06g, and the volume ratio of tetramethylethylenediamine and solvent is 0.005:1, and the amount of terephthalaldehyde is every 1mL solvent 0.02g.
(2) preparation of chitosan microball
The temperature regulating electrostatic spinning machine is 40 DEG C, open recirculating air button simultaneously, make the temperature in electrostatic spinning machine and remain on 40 DEG C, the container being loaded with jetting fluid is placed in the space of electrostatic spinning machine, when the temperature of liquid to be sprayed reaches 40 DEG C, jetting fluid to be loaded in the syringe 2 of electrostatic spinning machine and to push metal needle by syringe pump 1, the voltage of 5KV ~ 6KV is applied at metal needle place, namely jetting fluid forms liquid droplets, adopt fill receiving liquid container reception described in liquid droplets adopt magnetic stirring apparatus to stir receiving liquid with the rotating speed of 200rpm, shitosan after 8h in liquid droplets and the terephthalaldehyde in receiving liquid are completely crosslinked, obtain chitosan microball, the flow of described jetting fluid is 110 μ l/h, and the internal diameter of metal needle is 104 μm (metal needle model is 32G), and jet length is 5cm.
(3) wash
Adopt the receiving liquid of washed with isopropyl alcohol step (2) gained chitosan microball removing microsphere surface, then with distilled water washing removing isopropyl alcohol.Fig. 5 is the fluorescence picture of microballoon prepared by the present embodiment, and as shown in Figure 5, uniform particle diameter, the sphericity of microballoon prepared by the present embodiment are good, and the coefficient of variation (CV) value of chitosan microball is 4%, and the particle diameter of chitosan microball is 7 μm.
Embodiment 4
In the present embodiment, the preparation method of chitosan microball is as follows:
(1) preparation of jetting fluid and receiving liquid
The preparation of jetting fluid: water soluble chitosan is dissolved and forms chitosan solution in deionized water, then regulate between pH value to 6.3 ~ 6.4 of chitosan solution with the sodium hydrate aqueous solution that concentration is 1mol/L, by centrifugal for the chitosan solution after adjust ph, use Filter paper filtering, namely form jetting fluid; The mass ratio of described water soluble chitosan and deionized water is 0.02:1;
The preparation of receiving liquid: poly-ricinoleic acid glyceride, tetramethylethylenediamine are mixed with solvent, then add terephthalaldehyde, is stirred to terephthalaldehyde and dissolves namely form receiving liquid completely; Described solvent is the mixed liquor of toluene and n-hexyl alcohol, the volume ratio of toluene and n-hexyl alcohol is 2:1, the amount of poly-ricinoleic acid glyceride is every 1mL solvent 0.04g, and the volume ratio of tetramethylethylenediamine and solvent is 0.007:1, and the amount of terephthalaldehyde is every 1mL solvent 0.02g.
(2) preparation of chitosan microball
The temperature regulating electrostatic spinning machine is 45 DEG C, open recirculating air button simultaneously, make the temperature in electrostatic spinning machine and remain on 45 DEG C, the container being loaded with jetting fluid is placed in the space of electrostatic spinning machine, when the temperature of liquid to be sprayed reaches 45 DEG C, jetting fluid to be loaded in the syringe 2 of electrostatic spinning machine and to push metal needle by syringe pump 1, the voltage of 5KV ~ 6KV is applied at metal needle place, namely jetting fluid forms liquid droplets, adopt fill receiving liquid container reception described in liquid droplets adopt magnetic stirring apparatus to stir receiving liquid with the rotating speed of 250rpm, shitosan after 8h in liquid droplets and the terephthalaldehyde in receiving liquid are completely crosslinked, obtain chitosan microball, the flow of described jetting fluid is 110 μ l/h, and the internal diameter of metal needle is 104 μm (metal needle model is 32G), and jet length is 5cm.
(3) wash
Adopt washed with isopropyl alcohol step (2) gained chitosan microball to remove the receiving liquid of microsphere surface, then spend deionized water removing isopropyl alcohol.The particle diameter of chitosan microball prepared by the present embodiment is 6 μm, and the coefficient of variation (CV) value of chitosan microball is 3%.
Embodiment 5
In the present embodiment, the preparation method of chitosan microball is as follows:
(1) preparation of jetting fluid and receiving liquid
The preparation of jetting fluid: water soluble chitosan is dissolved and forms chitosan solution in deionized water, then regulate between pH value to 6.4 ~ 6.5 of chitosan solution with the sodium hydrate aqueous solution that concentration is 2mol/L, by centrifugal for the chitosan solution after adjust ph, use Filter paper filtering, namely form jetting fluid; The mass ratio of described water soluble chitosan and deionized water is 0.02:1;
The preparation of receiving liquid: poly-ricinoleic acid glyceride, tetramethylethylenediamine are mixed with solvent, then add terephthalaldehyde, is stirred to terephthalaldehyde and dissolves namely form receiving liquid completely; Described solvent is the mixed liquor of toluene and n-hexyl alcohol, the volume ratio of toluene and n-hexyl alcohol is 2:1, the amount of poly-ricinoleic acid glyceride is every 1mL solvent 0.04g, and the volume ratio of tetramethylethylenediamine and solvent is 0.008:1, and the amount of terephthalaldehyde is every 1mL solvent 0.01g.
(2) preparation of chitosan microball
The temperature regulating electrostatic spinning machine is 45 DEG C, open recirculating air button simultaneously, make the temperature in electrostatic spinning machine and remain on 45 DEG C, the container being loaded with jetting fluid is placed in the space of electrostatic spinning machine, when the temperature of liquid to be sprayed reaches 45 DEG C, jetting fluid to be loaded in the syringe 2 of electrostatic spinning machine and to push metal needle by syringe pump 1, the voltage of 5KV ~ 6KV is applied at metal needle place, namely jetting fluid forms liquid droplets, adopt fill receiving liquid container reception described in liquid droplets adopt magnetic stirring apparatus to stir receiving liquid with the rotating speed of 200rpm, shitosan after 8h in liquid droplets and the terephthalaldehyde in receiving liquid are completely crosslinked, obtain chitosan microball, the flow of described jetting fluid is 110 μ l/h, and the internal diameter of metal needle is 104 μm (metal needle model is 32G), and jet length is 5cm.
(3) wash
Adopt the receiving liquid of washed with isopropyl alcohol step (2) gained chitosan microball removing microsphere surface, then spend deionized water removing isopropyl alcohol.The particle diameter of chitosan microball prepared by the present embodiment is 6 μm, and the coefficient of variation (CV) value of chitosan microball is 3%.
Embodiment 6
In the present embodiment, the preparation method of chitosan microball is as follows:
(1) preparation of jetting fluid and receiving liquid
The preparation of jetting fluid: water soluble chitosan is dissolved and forms chitosan solution in deionized water, then regulate between pH value to 6.3 ~ 6.4 of chitosan solution with the sodium hydrate aqueous solution that concentration is 1mol/L, by centrifugal for the chitosan solution after adjust ph, use Filter paper filtering, namely form jetting fluid; The mass ratio of described water soluble chitosan and deionized water is 0.02:1;
The preparation of receiving liquid: poly-ricinoleic acid glyceride, tetramethylethylenediamine are mixed with solvent, then add terephthalaldehyde, is stirred to terephthalaldehyde and dissolves namely form receiving liquid completely; Described solvent is the mixed liquor of toluene and n-hexyl alcohol, the volume ratio of toluene and n-hexyl alcohol is 2:1, the amount of poly-ricinoleic acid glyceride is every 1mL solvent 0.06g, and the volume ratio of tetramethylethylenediamine and solvent is 0.005:1, and the amount of terephthalaldehyde is every 1mL solvent 0.02g.
(2) preparation of chitosan microball
The temperature regulating electrostatic spinning machine is 35 DEG C, open recirculating air button simultaneously, make the temperature in electrostatic spinning machine and remain on 35 DEG C, the container being loaded with jetting fluid is placed in the space of electrostatic spinning machine, when the temperature of liquid to be sprayed reaches 35 DEG C, jetting fluid to be loaded in the syringe 2 of electrostatic spinning machine and to push metal needle by syringe pump 1, the voltage of 5KV ~ 6KV is applied at metal needle place, namely jetting fluid forms liquid droplets, adopt fill receiving liquid container reception described in liquid droplets adopt magnetic stirring apparatus to stir receiving liquid with the rotating speed of 300rpm, shitosan after 8h in liquid droplets and the terephthalaldehyde in receiving liquid are completely crosslinked, obtain chitosan microball, the flow of described jetting fluid is 140 μ l/h, and the internal diameter of metal needle is 104 μm (metal needle model is 32G), and jet length is 5cm.
(3) wash
Adopt the receiving liquid of washed with isopropyl alcohol step (2) gained chitosan microball removing microsphere surface, then spend deionized water removing isopropyl alcohol.The particle diameter of chitosan microball prepared by the present embodiment is 9 μm, and the coefficient of variation (CV) value of chitosan microball is 4%.
Embodiment 7
In the present embodiment, the preparation method of chitosan microball is as follows:
(1) preparation of jetting fluid and receiving liquid
The preparation of jetting fluid: water soluble chitosan is dissolved and forms chitosan solution in deionized water, then regulate between pH value to 6.3 ~ 6.4 of chitosan solution with the sodium hydrate aqueous solution that concentration is 2mol/L, by centrifugal for the chitosan solution after adjust ph, use Filter paper filtering, namely form jetting fluid; The mass ratio of described water soluble chitosan and deionized water is 0.02:1;
The preparation of receiving liquid: poly-ricinoleic acid glyceride, tetramethylethylenediamine are mixed with solvent, then add terephthalaldehyde, is stirred to terephthalaldehyde and dissolves namely form receiving liquid completely; Described solvent is the mixed liquor of toluene and n-hexyl alcohol, the volume ratio of toluene and n-hexyl alcohol is 2:1, the amount of poly-ricinoleic acid glyceride is every 1mL solvent 0.04g, and the volume ratio of tetramethylethylenediamine and solvent is 0.006:1, and the amount of terephthalaldehyde is every 1mL solvent 0.02g.
(2) preparation of chitosan microball
The temperature regulating electrostatic spinning machine is 25 DEG C, open recirculating air button simultaneously, the temperature in electrostatic spinning machine is made to remain on 25 DEG C, the container being loaded with jetting fluid is placed in the space of electrostatic spinning machine, when the temperature of liquid to be sprayed is 25 DEG C, jetting fluid to be loaded in the syringe 2 of electrostatic spinning machine and to push metal needle by syringe pump 1, the voltage of 5KV ~ 6KV is applied at metal needle place, namely jetting fluid forms liquid droplets, adopt fill receiving liquid container reception described in liquid droplets adopt magnetic stirring apparatus to stir receiving liquid with the rotating speed of 200rpm, shitosan after 8h in liquid droplets and the terephthalaldehyde in receiving liquid are completely crosslinked, obtain chitosan microball, the flow of described jetting fluid is 140 μ l/h, and the internal diameter of metal needle is 104 μm (metal needle model is 32G), and jet length is 5cm.
(3) wash
Adopt the receiving liquid of washed with isopropyl alcohol step (2) gained chitosan microball removing microsphere surface, then spend deionized water removing isopropyl alcohol.The particle diameter of chitosan microball prepared by the present embodiment is 9 μm, and the coefficient of variation (CV) value of chitosan microball is 4%.
Embodiment 8
In the present embodiment, the preparation method of chitosan microball is as follows:
(1) preparation of jetting fluid and receiving liquid
The preparation of jetting fluid: water soluble chitosan is dissolved and forms chitosan solution in deionized water, then regulate between pH value to 6.3 ~ 6.4 of chitosan solution with the sodium hydrate aqueous solution that concentration is 2mol/L, by centrifugal for the chitosan solution after adjust ph, use Filter paper filtering, namely form jetting fluid; The mass ratio of described water soluble chitosan and deionized water is 0.02:1;
The preparation of receiving liquid: poly-ricinoleic acid glyceride, tetramethylethylenediamine are mixed with solvent, then add terephthalaldehyde, is stirred to terephthalaldehyde and dissolves namely form receiving liquid completely; Described solvent is the mixed liquor of toluene and n-hexyl alcohol, the volume ratio of toluene and n-hexyl alcohol is 1:1, the amount of poly-ricinoleic acid glyceride is every 1mL solvent 0.04g, and the volume ratio of tetramethylethylenediamine and solvent is 0.007:1, and the amount of terephthalaldehyde is every 1mL solvent 0.02g.
(2) preparation of chitosan microball
The temperature regulating electrostatic spinning machine is 25 DEG C, open recirculating air button simultaneously, the temperature in electrostatic spinning machine is made to remain on 25 DEG C, the container being loaded with jetting fluid is placed in the space of electrostatic spinning machine, when the temperature of liquid to be sprayed is 25 DEG C, jetting fluid to be loaded in the syringe 2 of electrostatic spinning machine and to push metal needle by syringe pump 1, the voltage of 5KV ~ 6KV is applied at metal needle place, namely jetting fluid forms liquid droplets, adopt fill receiving liquid container reception described in liquid droplets adopt magnetic stirring apparatus to stir receiving liquid with the rotating speed of 200rpm, shitosan after 8h in liquid droplets and the terephthalaldehyde in receiving liquid are completely crosslinked, obtain chitosan microball, the flow of described jetting fluid is 140 μ l/h, and the internal diameter of metal needle is 104 μm (metal needle model is 32G), and jet length is 8cm.
(3) wash
Adopt the receiving liquid of washed with isopropyl alcohol step (2) gained chitosan microball removing microsphere surface, then spend deionized water removing isopropyl alcohol.The particle diameter of chitosan microball prepared by the present embodiment is 11 μm, and the coefficient of variation (CV) value of chitosan microball is 5%.
Embodiment 9
In the present embodiment, the preparation method of chitosan microball is as follows:
(1) preparation of jetting fluid and receiving liquid
The preparation of jetting fluid: water soluble chitosan is dissolved and forms chitosan solution in deionized water, then pH value to 6.4 ~ 6.5 of chitosan solution are regulated with the sodium hydrate aqueous solution that concentration is 2mol/L, by centrifugal for the chitosan solution after adjust ph, use Filter paper filtering, namely form jetting fluid; The mass ratio of described water soluble chitosan and deionized water is 0.02:1;
The preparation of receiving liquid: poly-ricinoleic acid glyceride, tetramethylethylenediamine are mixed with solvent, then add terephthalaldehyde, is stirred to terephthalaldehyde and dissolves namely form receiving liquid completely; Described solvent is the mixed liquor of toluene and n-hexyl alcohol, the volume ratio of toluene and n-hexyl alcohol is 1:1, the amount of poly-ricinoleic acid glyceride is every 1mL solvent 0.06g, and the volume ratio of tetramethylethylenediamine and solvent is 0.008:1, and the amount of terephthalaldehyde is every 1mL solvent 0.02g.
(2) preparation of chitosan microball
The temperature regulating electrostatic spinning machine is 25 DEG C, open recirculating air button simultaneously, the temperature in electrostatic spinning machine is made to remain on 25 DEG C, the container being loaded with jetting fluid is placed in the space of electrostatic spinning machine, when the temperature of liquid to be sprayed is 25 DEG C, jetting fluid to be loaded in the syringe 2 of electrostatic spinning machine and to push metal needle by syringe pump 1, the voltage of 5KV ~ 6KV is applied at metal needle place, namely jetting fluid forms liquid droplets, adopt fill receiving liquid container reception described in liquid droplets adopt magnetic stirring apparatus to stir receiving liquid with the rotating speed of 200rpm, shitosan after 8h in liquid droplets and the terephthalaldehyde in receiving liquid are completely crosslinked, obtain chitosan microball, the flow of described jetting fluid is 140 μ l/h, and the internal diameter of metal needle is 104 μm (metal needle model is 32G), and jet length is 10cm.
(3) wash
Adopt the receiving liquid of washed with isopropyl alcohol step (2) gained chitosan microball removing microsphere surface, then spend deionized water removing isopropyl alcohol.The particle diameter of chitosan microball prepared by the present embodiment is 11 μm.
Embodiment 10
In the present embodiment, the preparation method of chitosan microball is as follows:
(1) preparation of jetting fluid and receiving liquid
The preparation of jetting fluid: water soluble chitosan is dissolved and forms chitosan solution in deionized water, then pH value to 6.4 ~ 6.5 of chitosan solution are regulated with the sodium hydrate aqueous solution that concentration is 2mol/L, by centrifugal for the chitosan solution after adjust ph, use Filter paper filtering, namely form jetting fluid; The mass ratio of described water soluble chitosan and deionized water is 0.02:1;
The preparation of receiving liquid: poly-ricinoleic acid glyceride, tetramethylethylenediamine are mixed with solvent, then add terephthalaldehyde, is stirred to terephthalaldehyde and dissolves namely form receiving liquid completely; Described solvent is the mixed liquor of toluene and n-hexyl alcohol, the volume ratio of toluene and n-hexyl alcohol is 1:1, the amount of poly-ricinoleic acid glyceride is every 1mL solvent 0.08g, and the volume ratio of tetramethylethylenediamine and solvent is 0.006:1, and the amount of terephthalaldehyde is every 1mL solvent 0.01g.
(2) preparation of chitosan microball
The temperature regulating electrostatic spinning machine is 45 DEG C, open recirculating air button simultaneously, make the temperature in electrostatic spinning machine and remain on 45 DEG C, the container being loaded with jetting fluid is placed in the space of electrostatic spinning machine, when the temperature of liquid to be sprayed reaches 45 DEG C, jetting fluid to be loaded in the syringe 2 of electrostatic spinning machine and to push metal needle by syringe pump 1, the voltage of 5KV ~ 6KV is applied at metal needle place, namely jetting fluid forms liquid droplets, adopt fill receiving liquid container reception described in liquid droplets adopt magnetic stirring apparatus to stir receiving liquid with the rotating speed of 300rpm, shitosan after 8h in liquid droplets and the terephthalaldehyde in receiving liquid are completely crosslinked, obtain chitosan microball, the flow of described jetting fluid is 110 μ l/h, and the internal diameter of metal needle is 104 μm (metal needle model is 32G), and jet length is 5cm.
(3) wash
Adopt washed with isopropyl alcohol step (2) gained chitosan microball, to remove the receiving liquid of microsphere surface, then spend deionized water removing isopropyl alcohol.The mean grain size of chitosan microball prepared by the present embodiment is 6.4 μm, and the coefficient of variation (CV) value of chitosan microball is 2.7%.
The damping fluid that pH value is 3.5 is put at 25 DEG C of chitosan microballs prepared by the present embodiment, its course of dissolution is observed under Laser Scanning Confocal Microscope, as shown in Figure 6, as shown in Figure 6, chitosan microball first occurs swelling, then dissolve gradually, when putting into pH value and being the 60s of the damping fluid of 3.5, chitosan microball dissolves completely.
Claims (10)
1. a solubility in acid chitosan microball, is characterized in that described microballoon is formed by shitosan and terephthalaldehyde cross-linking reaction, and the particle diameter of microballoon is 6 ~ 11 μm, and the value for coefficient of variation of microballoon is 2.7% ~ 5%.
2. the preparation method of solubility in acid chitosan microball described in claim 1, is characterized in that processing step is as follows:
(1) preparation of jetting fluid and receiving liquid
The preparation of jetting fluid: be dissolved in water by water soluble chitosan and form chitosan solution, then regulates pH value to 6.3 ~ 6.5 of chitosan solution with sodium hydrate aqueous solution, by centrifugal for the chitosan solution after adjust ph, filtration, namely form jetting fluid; The mass ratio of described water soluble chitosan and water is (0.02 ~ 0.03): 1;
The preparation of receiving liquid: poly-ricinoleic acid glyceride, tetramethylethylenediamine are mixed with solvent, then add terephthalaldehyde, is stirred to terephthalaldehyde and dissolves namely form receiving liquid completely; Described solvent is the mixed liquor of toluene and n-hexyl alcohol, the volume ratio of toluene and n-hexyl alcohol is (1 ~ 2): 1, the amount of poly-ricinoleic acid glyceride is every 1mL solvent 0.04 ~ 0.08g, the volume ratio of tetramethylethylenediamine and solvent is (0.004 ~ 0.008): 1, and the amount of terephthalaldehyde is every 1mL solvent 0.01 ~ 0.02g;
(2) preparation of chitosan microball
In the isoperibol of 25 ~ 45 DEG C, the jetting fluid identical with isoperibol temperature is added in the syringe of electrospinning device, then metal needle is pushed by syringe pump, high-pressure electrostatic is applied at metal needle place, namely jetting fluid forms liquid droplets, adopt the container filling receiving liquid to receive described liquid droplets under agitation, the shitosan in drop to be sprayed and the terephthalaldehyde in receiving liquid are completely crosslinked, obtain chitosan microball; The voltage controlling to be applied to metal needle place is 5 ~ 6kV, and jet length is 5 ~ 10cm, and the internal diameter of metal needle is 104 ~ 130 μm, and the flow of jetting fluid is 110 ~ 140 μ L/h;
(3) wash
Adopt washed with isopropyl alcohol step (2) gained chitosan microball to remove the receiving liquid of microsphere surface, then wash removing isopropyl alcohol with water.
3. the preparation method of solubility in acid chitosan microball according to claim 2, is characterized in that stirring receiving liquid with the rotating speed of 200 ~ 300rpm in step (2).
4. the preparation method of solubility in acid chitosan microball according to Claims 2 or 3, is characterized in that weight-average molecular weight≤5000 of described water soluble chitosan.
5. the preparation method of solubility in acid chitosan microball according to Claims 2 or 3, is characterized in that described water is deionized water or distilled water.
6. the preparation method of solubility in acid chitosan microball according to claim 4, is characterized in that described water is deionized water or distilled water.
7. the preparation method of solubility in acid chitosan microball according to Claims 2 or 3, is characterized in that the concentration of described sodium hydrate aqueous solution is at least 1mol/L.
8. the preparation method of solubility in acid chitosan microball according to claim 4, is characterized in that the concentration of described sodium hydrate aqueous solution is at least 1mol/L.
9. the preparation method of solubility in acid chitosan microball according to claim 5, is characterized in that the concentration of described sodium hydrate aqueous solution is at least 1mol/L.
10. the preparation method of solubility in acid chitosan microball according to claim 6, is characterized in that the concentration of described sodium hydrate aqueous solution is at least 1mol/L.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105106967A (en) * | 2015-08-07 | 2015-12-02 | 四川大学 | Chitosan microparticles with shapes and sizes identical to lurid cells and preparation method of chitosan microparticles |
| CN107118286A (en) * | 2017-04-27 | 2017-09-01 | 广西科学院 | The preparation method of chitosan crosslinked 2,5 furans dicarbaldehyde hydrogels |
| CN112617179A (en) * | 2020-11-13 | 2021-04-09 | 华南理工大学 | High-resistance single-particle starch microsphere, and preparation method and application based on electrostatic spraying method |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6096344A (en) * | 1995-07-28 | 2000-08-01 | Advanced Polymer Systems, Inc. | Bioerodible porous compositions |
| WO2006024490A2 (en) * | 2004-08-30 | 2006-03-09 | Interstitial Therapeutics | Methods and compositions for the treatment of cell proliferation |
| CN101148511A (en) * | 2006-09-18 | 2008-03-26 | 中国科学院过程工程研究所 | Method for preparing fluorescent chitosan micro-sphere and application of the same in tracer field |
| CN102114006A (en) * | 2011-02-24 | 2011-07-06 | 四川大学 | Preparation method of chitosan hollow and core-shell microcapsule with pH-sensitive burst release property |
| CN102626602A (en) * | 2012-03-31 | 2012-08-08 | 四川大学 | Method for preparing chitosan micro-capsule with single emulsion serving as formwork |
| US20130123314A1 (en) * | 2009-02-20 | 2013-05-16 | Academic Hospital Maastricht | Method for improving cartilage repair and/or preventing cartilage degeneration in a joint |
| CN103962075A (en) * | 2014-05-19 | 2014-08-06 | 四川大学 | Preparation method of composite microcapsule with monodisperse capsule-in-capsule structure |
-
2015
- 2015-03-27 CN CN201510142199.6A patent/CN104792688B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6096344A (en) * | 1995-07-28 | 2000-08-01 | Advanced Polymer Systems, Inc. | Bioerodible porous compositions |
| WO2006024490A2 (en) * | 2004-08-30 | 2006-03-09 | Interstitial Therapeutics | Methods and compositions for the treatment of cell proliferation |
| CN101148511A (en) * | 2006-09-18 | 2008-03-26 | 中国科学院过程工程研究所 | Method for preparing fluorescent chitosan micro-sphere and application of the same in tracer field |
| US20130123314A1 (en) * | 2009-02-20 | 2013-05-16 | Academic Hospital Maastricht | Method for improving cartilage repair and/or preventing cartilage degeneration in a joint |
| CN102114006A (en) * | 2011-02-24 | 2011-07-06 | 四川大学 | Preparation method of chitosan hollow and core-shell microcapsule with pH-sensitive burst release property |
| CN102626602A (en) * | 2012-03-31 | 2012-08-08 | 四川大学 | Method for preparing chitosan micro-capsule with single emulsion serving as formwork |
| CN103962075A (en) * | 2014-05-19 | 2014-08-06 | 四川大学 | Preparation method of composite microcapsule with monodisperse capsule-in-capsule structure |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105106967A (en) * | 2015-08-07 | 2015-12-02 | 四川大学 | Chitosan microparticles with shapes and sizes identical to lurid cells and preparation method of chitosan microparticles |
| CN105106967B (en) * | 2015-08-07 | 2017-12-15 | 四川大学 | Chitosan microparticle with erythrocyte shapes and sizes and preparation method thereof |
| CN107118286A (en) * | 2017-04-27 | 2017-09-01 | 广西科学院 | The preparation method of chitosan crosslinked 2,5 furans dicarbaldehyde hydrogels |
| CN107118286B (en) * | 2017-04-27 | 2019-05-10 | 广西科学院 | The preparation method of chitosan crosslinked 2,5-furandaldehyde hydrogel |
| CN112617179A (en) * | 2020-11-13 | 2021-04-09 | 华南理工大学 | High-resistance single-particle starch microsphere, and preparation method and application based on electrostatic spraying method |
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