CN109350609A - A kind of preparation of chitosan package zeins nanoparticle and the application for loading acacetin sodium salt - Google Patents
A kind of preparation of chitosan package zeins nanoparticle and the application for loading acacetin sodium salt Download PDFInfo
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- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
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Abstract
The invention discloses a kind of preparation method and applications of chitosan package zein nanoparticle.The nanoparticle the preparation method comprises the following steps: spray bottle (about 10~50 μm of mist droplet particle size) is added in the ethanol solution of zein, another spray bottle (about 10~50 μm of mist droplet particle size) separately is added in water phase, by two spray bottles to the solution sprayed up to the nanoparticle containing zein in closed device, the partial size of zein nanoparticle about 160~200nm, -8~12mv of zeta current potential.Suitable zein nanoparticle solution is added in chitosan solution, the nanoparticle of chitosan package zein can be obtained through being incubated for, the partial size that chitosan wraps up zein nanoparticle is 160~180nm, zeta current potential about 50mv.The present invention can continuous production chitosan wrap up zein nanoparticle, preparation method simplicity is pollution-free, mild safety, yield are high.When for loading acacetin sodium salt, drugloading rate is about 20~30%, 190~260nm of partial size, zeta 35~45mv of current potential, its stability can be improved after being wrapped up by the nanoparticle in acacetin sodium salt.
Description
Technical field
The present invention relates to biological medicine nano controlled-release technical fields, and in particular to a kind of chitosan package zeins
The preparation method of nanoparticle and the application for loading acacetin sodium salt.
Background technique
Zeins is main Storage protein in corn, accounts for the 35~60% of endosperm gross protein, average mark
Son amount about 40kD.Due to containing a large amount of hydrophobic amino acid in zeins, such as glutamine and proline；Lack energy
The acidity of electrification, basic amino acid, especially shortage tryptophan and lysine；Containing more sulfur-containing amino acid, due to its amino
Acid composition is unbalanced, and nutritive value is not high.Recent study discovery hydrophilic segment and is dredged in zeins molecular structure
Water section subregion is obvious, has unique self assembly characteristic, film forming, gelation, has water resistant, grease resistance and good life
Object compatibility, bioadhesive.Based on the amphiphilic of zein, zein can conveniently be obtained by classical phase separation method
The nanoparticle for encapsulating hydrophobic drug, but since the difficulty that molecular scale finely manipulates is larger, cause the reproducibility of method compared with
Difference, process are not easy to control, and the Zein nanoparticle stability that nanoparticle particle diameter distribution is wider, simple is poor, is easy to
Aggregation occurs and precipitates.
Chitosan is one of natural, reproducible natural polymer, is obtained by chitin by deacetylation,
With excellent physics, chemical property, good biocompatibility, biodegradable, in cell culture, drug controlled release, albumen
The numerous areas such as matter separation are widely used.The molecular weight of chitosan has significant impact to its solubility and physicochemical property.Make
For natural unique alkaline polysaccharide, it is soluble in acid solution and with positive electricity, thus with electronegative grain
Son, which be combined with each other to play by electrostatic interaction, to be increased nanoparticle stability, reduces burst drug release, and target cell is improved
The effects of to the endocytosis of nanoparticle, intake.
Acacetin is a kind of flavone compound found in Herba Saussureae Involueratae first, it was reported that has extensive pharmacology
Activity, such as it is anti-inflammatory, antitumor, anti-oxidant.A series of research discovery acacetins can be blocked by multiple ion channels recently
Effect is used for the treatment of atrial fibrillation, is a kind of anti-arrhythmia candidate compound of great potential, but since its water solubility is very poor,
Limit its clinical application.It can be improved thousands of times by the sodium salt water solubility of phosphate connection preparation, and can be in body
Hydrolysis occurs under the action of interior phosphate and generates acacetin, plays the therapeutic effect of acacetin itself, can be used as prodrug
It uses.But the chemical stability of acacetin sodium salt is not good enough, and it is more sensitive to high humidity, high temperature, it is wrapped in nanoparticle
In, its stability can be improved.
Summary of the invention
The first purpose of the invention is to provide a kind of preparation methods of chitosan package zeins nanoparticle, should
Method prepares zeins nanoparticle first with relatively controllable phase separation method, secondly using chitosan in acid solution
It is middle institute it is positively charged, with zeins nanoparticle by electrostatic interaction in conjunction with, it is molten to prepare Chitosan-coated corn alcohol
Protein nano grain improves the reproducibility of preparation to increase the stability of zein nanoparticle.
A second object of the present invention is to provide the applications of the drug delivery system.
First purpose to realize the present invention, technical solution be the following steps are included:
(1) zeins is dissolved into ethanol solution, is added in spray bottle；
(2) separately the water phase of deionized water or the appropriate stabilizer of addition is added in another spray bottle；By step (1) and step
(2) two spray bottles described in are in closed device to the mixed solution sprayed up to the nanoparticle containing zeins.By institute
It obtains solution rotating and evaporates 30min, remove ethyl alcohol.
(3) zeins nanoparticle solution described in step (3) is added in chitosan solution, agitated incubation
Afterwards, 10min is centrifuged by 2000rpm and takes supernatant, then 30min is centrifuged by 12000rpm, collect sediment to get poly- to shell
Sugar package zeins nanoparticle.
(4) when being applied to load acacetin sodium salt, acacetin sodium salt is dissolved in the deionized water of step (4) or is added
In the water phase for entering appropriate stabilizer.
The concentration of zeins described in step (1) is 5~20mg/ml.
Concentration of alcohol described in step (1) is 70~80%.
Solution described in step (1) and water phase are entered in closed device with the speed ratio of 1:1~1:3.
The mass volume ratio of stabilizer described in step (2) and water is 1~10%.
Two spray bottles obtain milky zeins nanoparticle solution to spray in closed device in step (3).
The mass ratio of chitosan described in step (4) and zeins is 1~2:1.
The solvent of chitosan solution described in step (4) is 1~10% acetum.
Mixing speed described in step (4) is 200~600rpm.
Incubation time described in step (4) is 10~30min.
The concentration of acacetin sodium salt described in step (5) is 1~4mg/ml.
Chitosan package zeins nanoparticle partial size obtained by the above method is 190~250nm (PDI=0.10
~0.20), zeta current potential is about 50mv；It is 20~30% for drugloading rate when loading acacetin sodium salt, partial size about 190~
260nm, zeta current potential about 35~45mv.
Compared with prior art, the present invention has the following advantages and beneficial effects:
(1) Chitosan-coated zein nanoparticle can be continuously prepared, stability is better than zein nanoparticle.
(2) zein nanoparticle yield is higher than the methods of common phase separation method and stirring evaporation, and preparation condition is mild, receives
Grain of rice yield is higher, and particle diameter distribution is uniform, and dispersibility is high in water, to entrapment efficiency height.
(3) pass through control corn protein solution concentration, aqueous-phase concentration, the jet velocity of two-phase, chitosan concentration, chitosan
The parameters such as molecular weight effectively control the granule size of final product.
Detailed description of the invention
The particle diameter distribution and potential diagram of Fig. 1 zeins nanoparticle；
The grain size distribution of the PEG400 of Fig. 2 5% stable zeins nanoparticle；
The grain size distribution of the F68 of Fig. 3 5% stable zeins nanoparticle；
The particle diameter distribution and potential diagram of Fig. 4 chitosan (MW 5,000) cladding zeins nanoparticle；
The particle diameter distribution and potential diagram of Fig. 5 chitosan (MW 200,000) cladding zeins nanoparticle；
Fig. 6 loads the particle diameter distribution and potential diagram of the Chitosan-coated zeins nanoparticle of acacetin；
Specific embodiment
Below with reference to embodiment, the preferred embodiments of the present invention will be described in detail.Following embodiment is for illustrating this
Invention, but be not intended to limit the scope of the invention.Each raw material of following embodiment is conventional commercial product.
Embodiment 1
(1) precision weighs 100.1mg zeins and is dissolved in 80% ethyl alcohol of 10ml, and ultrasonic 5min keeps it completely molten
Solution, is placed in the spray bottle of 50ml；
(2) 10ml deionized water is measured, is placed in the spray bottle of another 50ml；
(3) at room temperature, by two spray bottles in closed device to spray, obtain opalescent solution.By acquired solution
The rotary evaporation 30min at 50 DEG C removes ethyl alcohol；
(4) being centrifuged 10min by 2000rpm takes supernatant as sample, with nano-zs90 Malvern partial size and potentiometer
The partial size and current potential of product are measured, average grain diameter is about (polydispersity index=0.209) 180.5nm, and current potential is -11.9mv, is received
The partial size and potential image of the grain of rice are shown in attached drawing 1.
Embodiment 2
(1) precision weighs 100.3mg zeins and is dissolved in 80% ethyl alcohol of 10ml, and ultrasonic 5min keeps it completely molten
Solution, is placed in the spray bottle of 50ml；
(2) precision weighs 5.2mg PEG400 and is dissolved in 10ml deionized water, and ultrasonic 5min makes it completely dissolved, and is placed in another
In the spray bottle of one 50ml；
(3) at room temperature, by two spray bottles in closed device to spray, obtain opalescent solution.By acquired solution
The rotary evaporation 30min at 50 DEG C removes ethyl alcohol；
(4) being centrifuged 10min by 2000rpm takes supernatant as sample, with nano-zs90 Malvern partial size and potentiometer
Measure the partial size of product.The average grain diameter of 5% PEG400 stable zeins nanoparticle is about (more points of 159.3nm
Dissipate index=0.186), grain size distribution is shown in Fig. 2.
Embodiment 3
(1) precision weighs 100.3mg zeins and is dissolved in 80% ethyl alcohol of 10ml, and ultrasonic 5min keeps it completely molten
Solution, is placed in the spray bottle of 50ml；
(2) precision weighs 5.3mg F68 and is dissolved in 10ml deionized water, and ultrasonic 5min makes it completely dissolved, and is placed in another
In the spray bottle of 50ml；
(3) at room temperature, by two spray bottles in closed device to spray, obtain opalescent solution.By acquired solution
The rotary evaporation 30min at 50 DEG C removes ethyl alcohol；
(4) being centrifuged 10min by 2000rpm takes supernatant as sample, with nano-zs90 Malvern partial size and potentiometer
Measure product partial size, the partial size of 5% F68 stable zeins nanoparticle be about 246.7nm (polydispersity index=
0.224), grain size distribution is shown in Fig. 3.
Embodiment 4
(1) precision weighs 100.2mg zeins and is dissolved in 80% ethyl alcohol of 10ml, and ultrasonic 5min keeps it completely molten
Solution, is placed in the spray bottle of 50ml；
(2) 10ml deionized water is measured, is placed in the spray bottle of another 50ml；
(3) at room temperature, by two spray bottles in closed device to spray, obtain opalescent solution.By acquired solution
The rotary evaporation 30min at 50 DEG C removes ethyl alcohol；
(4) the accurate chitosan oligosaccharide (MW 5,000) for weighing 6.8mg, is added in the acetum of 10ml 1%.With 600rpm
Speed stirring, take the solution 1ml in step 3 to be slowly dropped into, stir 30min；
(5) being centrifuged 10min by 2000rpm takes supernatant as sample, with nano-zs90 Malvern partial size and potentiometer
Measure the partial size and current potential of product, average grain diameter is about (polydispersity index=0.187) 166nm, current potential 52.5mv, partial size and
Potential image is shown in Fig. 4.
Embodiment 5
(1) precision weighs 100.4mg zeins and is dissolved in 80% ethyl alcohol of 10ml, and ultrasonic 5min keeps it completely molten
Solution, is placed in the spray bottle of 50ml；
(2) 10ml deionized water is measured, is placed in the spray bottle of another 50ml；
(3) at room temperature, by two spray bottles in closed device to spray, obtain opalescent solution.By acquired solution
The rotary evaporation 30min at 50 DEG C removes ethyl alcohol；
(4) the accurate chitosan (MW 20,000) for weighing 6.9mg is added in 1% acetum of 10ml.With 600rpm
Speed stirring, take the solution in 1ml step 3 to be slowly dropped into, stir 30min；
(5) 10min is centrifuged by 2000rpm and discards lower sediment, measured with nano-zs90 Malvern partial size and potentiometer
The partial size and current potential of product, average grain diameter are about (polydispersity index=0.317) 221.5nm, current potential 54.7mv, partial size and electricity
Bit distribution figure is shown in Fig. 5.
Embodiment 6
(1) precision weighs 100mg zeins and is dissolved in 80% ethyl alcohol of 10ml, and ultrasonic 5min keeps it completely molten
Solution, is placed in the spray bottle of 50ml；
(2) precision weighs 20mg acacetin sodium salt and is dissolved in 10ml deionized water, and ultrasonic 5min makes it completely dissolved, and sets
In the spray bottle of another 50ml；
(3) at room temperature, by two spray bottles in closed device to spray, obtain opalescent solution.By acquired solution
The rotary evaporation 30min at 50 DEG C removes ethyl alcohol；
(4) being centrifuged 10min by 2000rpm takes supernatant as sample, sampling nano-zs90 Malvern partial size and electricity
The partial size and current potential of position instrument measurement nanoparticle, average grain diameter are about 200.6nm, and current potential is -19.6mv；
(5) it by the chitosan oligosaccharide of 13.8mg (MW 5,000), is added in 1% acetum of 20ml.With the speed of 600rpm
Degree stirring, takes the sample solution in 2ml step 4 to be slowly dropped into, and stirs 30min；
(6) being centrifuged 10min by 2000rpm takes supernatant as sample, with nano-zs90 Malvern partial size and potentiometer
The partial size and current potential of nanoparticle are measured, average grain diameter is about (polydispersity index=0.374) 245.7nm, and current potential 38.9mv leads to
Cross calculation formula: drugloading rate=(the free dose of total dose -)/drug-carrying nanometer particle gross weight, obtaining drugloading rate is 29.2%, grain
Diameter and potential image are shown in Fig. 6.
Claims (10)
1. a kind of preparation method of chitosan package zeins nanoparticle, it is characterised in that:
[1] zeins is dissolved into ethanol solution by, is added in spray bottle；
[2] water phase of deionized water or the appropriate stabilizer of addition is separately added in another spray bottle by；
[3] by two spray bottles described in step (1) and step (2) in closed device to spraying up to receiving containing zein
The mixed solution of the grain of rice.
[4] zeins nanoparticle solution described in step (3) is added in chitosan solution, after agitated incubation,
Chitosan is obtained by centrifugal purification wraps up zeins nanoparticle.
2. the preparation method of Zein nanoparticle according to claim 1, which is characterized in that the ethyl alcohol is molten
The concentration of liquid is 70~80%.
3. the preparation method of Zein nanoparticle according to claim 1, which is characterized in that the corn alcohol
The mass concentration of molten protein solution is 5~20mg/ml, and the molecular weight of zeins is 25~45kDa.
4. the preparation method of Zein nanoparticle according to claim 1, which is characterized in that the stabilizer
Type be one or more of F68, F127, Span80, RH40, PEG400, Pu Luoshamu 407.
5. the preparation method of Zein nanoparticle according to claim 1, which is characterized in that the stabilizer
Mass volume ratio with water is 1~10%.
6. the preparation method of Zein nanoparticle according to claim 1, which is characterized in that the alcoholic solution
Entered in closed device with water phase with the speed ratio of 1:1~1:3.
7. the preparation method of Zein nanoparticle according to claim 1, which is characterized in that the chitosan
Molecular weight be 5~20kDa.
8. the preparation method of Zein nanoparticle according to claim 1, which is characterized in that the chitosan
Mass ratio with zeins is 1~2:1.
9. the preparation method of Zein nanoparticle according to claim 1, which is characterized in that the chitosan
Mixing speed with zeins nanoparticle is 200~600rpm.
10. the preparation method of Zein nanoparticle according to claim 1, which is characterized in that the shell is poly-
The incubation time of sugar and zeins nanoparticle is 10~30min.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109770333A (en) * | 2019-03-18 | 2019-05-21 | 青岛农业大学 | A kind of alcohol soluble protein zymolyte nanoparticle and its preparation method and application of modification of chitosan modification |
| CN111228420A (en) * | 2020-02-27 | 2020-06-05 | 佛山科学技术学院 | Preparation method of coix seed oil nano composite particles |
| CN112205628A (en) * | 2020-09-18 | 2021-01-12 | 广州大学 | Composite condensate with double embedding functions and preparation method and application thereof |
| CN112245409A (en) * | 2020-10-23 | 2021-01-22 | 安徽大学 | Vegetable protein-ursodesoxycholic acid sustained-release nanoparticle composite microcapsule and preparation method thereof |
| WO2021231658A1 (en) * | 2020-05-15 | 2021-11-18 | Elc Management Llc | Shear-thinning cosmetic composition |
| CN114129538A (en) * | 2021-10-26 | 2022-03-04 | 浙江科技学院 | Preparation method and application of crocin-loaded zein nanoparticles |
| CN114568674A (en) * | 2022-02-17 | 2022-06-03 | 江南大学 | Preparation method of high-embedding-rate microcapsules for powdering spice essential oil |
| CN116172903A (en) * | 2023-03-13 | 2023-05-30 | 北京工商大学 | Corn protein polysaccharide nanoparticle and preparation method and application thereof |
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Cited By (13)
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|---|---|---|---|---|
| CN109770333A (en) * | 2019-03-18 | 2019-05-21 | 青岛农业大学 | A kind of alcohol soluble protein zymolyte nanoparticle and its preparation method and application of modification of chitosan modification |
| CN111228420A (en) * | 2020-02-27 | 2020-06-05 | 佛山科学技术学院 | Preparation method of coix seed oil nano composite particles |
| US11446229B2 (en) | 2020-05-15 | 2022-09-20 | Elc Management Llc | Shear-thinning cosmetic composition |
| CN115697281B (en) * | 2020-05-15 | 2024-04-05 | Elc管理有限责任公司 | Shear thinning cosmetic composition |
| WO2021231658A1 (en) * | 2020-05-15 | 2021-11-18 | Elc Management Llc | Shear-thinning cosmetic composition |
| CN115697281A (en) * | 2020-05-15 | 2023-02-03 | Elc管理有限责任公司 | Shear thinning cosmetic composition |
| CN112205628A (en) * | 2020-09-18 | 2021-01-12 | 广州大学 | Composite condensate with double embedding functions and preparation method and application thereof |
| CN112245409B (en) * | 2020-10-23 | 2022-03-15 | 安徽大学 | Vegetable protein-ursodesoxycholic acid sustained-release nanoparticle composite microcapsule and preparation method thereof |
| CN112245409A (en) * | 2020-10-23 | 2021-01-22 | 安徽大学 | Vegetable protein-ursodesoxycholic acid sustained-release nanoparticle composite microcapsule and preparation method thereof |
| CN114129538B (en) * | 2021-10-26 | 2022-12-09 | 浙江科技学院 | Preparation method and application of crocin-loaded zein nanoparticles |
| CN114129538A (en) * | 2021-10-26 | 2022-03-04 | 浙江科技学院 | Preparation method and application of crocin-loaded zein nanoparticles |
| CN114568674A (en) * | 2022-02-17 | 2022-06-03 | 江南大学 | Preparation method of high-embedding-rate microcapsules for powdering spice essential oil |
| CN116172903A (en) * | 2023-03-13 | 2023-05-30 | 北京工商大学 | Corn protein polysaccharide nanoparticle and preparation method and application thereof |
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