CN114344282A - Nanoparticle with three-layer composite structure formed by prolamin, sodium caseinate and polysaccharide and rapid preparation method thereof - Google Patents
Nanoparticle with three-layer composite structure formed by prolamin, sodium caseinate and polysaccharide and rapid preparation method thereof Download PDFInfo
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Abstract
The invention provides a nanoparticle with a three-layer composite structure and a rapid preparation method thereof, wherein the nanoparticle has a three-layer structure and comprises an inner layer of alcohol soluble protein structure, a middle layer of sodium caseinate structure and an outer layer of polysaccharide structure, the inner layer of alcohol soluble protein structure is coated by the middle layer of sodium caseinate structure and the outer layer of polysaccharide structure together to form the three-layer composite nanoparticle structure, the preparation method comprises the steps of dissolving alcohol soluble protein in an organic phase, dissolving sodium caseinate and polysaccharide in water according to a certain proportion, and then adopting a nano-precipitation method to assemble the alcohol soluble protein, sodium caseinate and polysaccharide together to prepare stable nanoparticles, the method for preparing the composite nanoparticles has the characteristics of simple and convenient operation, good stability, high activity and suitability for large-scale production, can be used in the fields of food, medicine, cosmetics and health products.
Description
Technical Field
The invention relates to the technical field of protein compound nanoparticle preparation, in particular to a nanoparticle with a three-layer compound structure, which is formed by prolamin, sodium caseinate and polysaccharide and has a simple structure and a rapid preparation method.
Background
It is well known that more and more structurally functional compounds such as plant polyphenols have been found to have potential health benefits, particularly in ameliorating cardiovascular and cerebrovascular diseases, however, it is difficult to achieve laboratory-derived levels in practical applications, or in vivo levels do not yield results consistent with those obtained in vitro. Meanwhile, partial potential functional compounds have the defects of dark color, strong stimulation, low water solubility, unstable chemistry and the like, and the practical application of the functional compounds is limited by the problems. Physical effects the above problems in drug utilization are solved by embedding being the most potential method. Active substances are embedded in the formed dense mesh-shaped matrix in the form of spheres or capsules by the macromolecular micro-nano particles, so that the purposes of protecting the inactivation and delivery of the active substances are achieved. This has led to the need to synthesize different molecules to entrap functional compounds to mask irritation, improve their dispersibility, stability and bioavailability, etc. In view of this, the biocompatibility problems of the preparation materials (synthetic polymers or surfactants), residual organic solvents and acids, and any products degraded during the preparation process, need to be investigated. As a preventive strategy, for functional food applications only generally recognized safe ingredients can be used, of which vegetable proteins have received increasing attention as delivery materials.
The nanometer particle prepared by using the prolamin as the material has the functions of coating, protecting and delivering medicaments. Then the single prolamin nanoparticle has the characteristics of poor stability, low embedding rate and general slow-release effect. Prolamin nanoparticles with casein attached to the surface can be obtained by anti-solvent co-precipitation of another biopolymer such as sodium caseinate. The sodium caseinate used as the stabilizer has the characteristics of redispersibility and higher ion strength resistance. Since prolamin-based nanoparticles achieve stabilization mainly through charge repulsion, sodium-casein stabilized prolamin particles aggregate at pH = about 4.6 (isoelectric point of sodium-casein), which is not conducive to drug delivery. Polysaccharide is also widely reported as a prolamin nanoparticle stabilizer, and prolamin nanoparticles stabilized by polysaccharide do not have the characteristic of ion-resistant strength, so that the adoption of a sodium caseinate-polysaccharide composite stabilizer is expected to combine the advantages of the prolamin nanoparticle stabilizer and the prolamin nanoparticle stabilizer.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provide a nanoparticle with a three-layer composite structure, which is formed by prolamine, sodium caseinate and polysaccharide and has a simple structure and a rapid preparation method.
The technical scheme adopted by the invention for solving the technical problems is as follows:
a three-layer composite nanoparticle is formed by prolamin, sodium caseinate and polysaccharide, and is characterized in that the three-layer composite nanoparticle comprises an inner prolamin structure, a middle casein structure and an outer polysaccharide structure, wherein the middle casein structure and the outer polysaccharide structure jointly coat the inner prolamin structure to form a three-layer composite nanoparticle structure.
A preparation method of three-layer composite nano-particles formed by prolamin, sodium caseinate and polysaccharide is characterized in that the three-layer composite nano-particles are obtained by a one-step nano-precipitation method, and the preparation method specifically comprises the following steps:
(1) dissolving prolamin in organic phase, and stirring to completely dissolve;
(2) dissolving sodium caseinate and polysaccharide in water phase together, and stirring to dissolve completely;
(3) dropwise adding the prolamin organic phase obtained in the step (1) into the water phase obtained in the step (2) under the stirring condition to obtain dispersion liquid.
(4) And (4) carrying out rotary evaporation on the dispersion liquid obtained in the step (3) to remove an organic phase, centrifuging to remove large particles to obtain a nano particle dispersion liquid, and then carrying out vacuum freeze drying to obtain a nano particle solid.
The alcohol soluble protein is plant-extracted or conventional alcohol soluble protein sold in the market, and the concentration range of the alcohol soluble protein is 0.5-5% (w/v), and preferably 0.5-4%.
The alcohol soluble protein is dissolved by adopting an organic phase, and the organic phase comprises an ethanol aqueous solution, a methanol aqueous solution, an isopropanol aqueous solution, an acetone aqueous solution or ethylene glycol.
The volume fraction of the ethanol aqueous solution is 30-90%, the volume fraction of the methanol aqueous solution is 45-95%, the volume fraction of the isopropanol aqueous solution is 40-85%, preferably, the volume fraction of the organic phase methanol aqueous solution is 60-90% (w/v), the volume fraction of the ethanol aqueous solution is 65-85% (w/v), and the volume fraction of the isopropanol aqueous solution is 70-80% (v/v).
The sodium caseinate and the polysaccharide are dissolved in the water phase together according to a certain proportion, and the proportion range of the organic phase to the water phase is 1: 1.5-10, preferably, the volume of the organic phase and the aqueous phase is 1: 2-6 (v/v).
The polysaccharide type described in the present invention is a negative polysaccharide.
The concentration of the sodium caseinate is 0.15-10% (w/v), the concentration of the polysaccharide is 0.001-7% (w/v), the concentration of the sodium caseinate is 0.15-0.3% (w/v), and the concentration of the polysaccharide is 0.001-1% (w/v).
The nano-particles have a three-layer structure, namely, the prolamin structure at the inner layer is a hydrophobic core formed by prolamin, the sodium casein structure at the middle layer is an amphiphilic sodium casein monolayer, and the hydrophilic polysaccharide surface of the polysaccharide structure at the outer layer is attached to form a shell, the size of the nano-particles prepared by the method is changed along with the variety of the polysaccharide at the outer layer and is different from 50-600 nm, and the polydispersity is less than 0.3.
The alcohol-soluble protein is zein, millet alcohol-soluble protein, hordein and the like;
aiming at the problems of poor stability and low embedding rate of prolamin nanoparticles, one of the purposes of the patent is to prepare a stable nanoparticle for embedding a medicament, so that the stability targeting property and the bioavailability of the medicament are improved.
Aiming at the complex problem of preparing multilayer nano particles at present, the invention provides a one-step method for preparing nano particles with the structure, so that the complexity of the preparation process is greatly reduced.
The negative polysaccharide is polysaccharide with negative charge, such as chondroitin sulfate, sulfate radical modified curdlan, fucoidin, sodium alginate, sodium carboxymethylcellulose and the like; the invention utilizes the self-assembly property of alcohol soluble protein, utilizes the acting force between biomacromolecules and adopts an anti-solvent method to prepare the nano-particles with hydrophobic alcohol soluble protein cores, casein sodium single layers and hydrophilic shells. The method has the advantages that a novel and simple method for preparing the nano particles with the multilayer composite structure is provided, the size distribution of the particles formed at the same time is uniform, the embedding rate of lipid solubility is obviously improved, and the stability and the biological activity of the nano particles are enhanced. The three-layer stable nanoparticle provided by the invention can be used for protecting and delivering drugs or functional factors.
Drawings
FIG. 1 is a graph of surface potential as a function of pH for single-layered (naked prolamin protein), two-layered (sodium caseinate and chondroitin sulfate coated, respectively) and three-layered (casein and chondroitin sulfate co-coated) nanoparticles prepared in example 1.
FIG. 2 is a scanning electron microscope image of different magnification times of the triple-layered nanoparticles prepared in example 1.
FIG. 3 is a transmission electron micrograph of two-layered and three-layered nanoparticles of example 1.
Figure 4 is a graph of the pH stability of the three-layered nanoparticles of example 1.
FIG. 5 is a graph of the ionic strength stability of the three-layered nanoparticles of example 1.
Figure 6 is a plot of the nanoparticle size and polydispersity as freshly prepared and redispersed in example 1.
Fig. 7 is an infrared analysis chart of nanoparticles prepared in example 1.
Detailed Description
The invention is further described below with reference to the accompanying drawings:
as shown in the drawing, the preparation conditions for preparing the multilayer structure generally include two or more steps. According to the three-layer structure of the present patent, it is common practice to prepare casein-stabilized nanoparticles as a first step; and the second step is to put the nanoparticles stabilized by sodium caseinate into the polysaccharide aqueous solution to form three-layer composite nanoparticles. Or in the preparation, the sodium caseinate and the polysaccharide are usually dissolved separately and then the organic phase and one of the aqueous phases are added dropwise to the other aqueous phase. But either method creates complexity in the manufacturing process. The invention prepares the prolamin nanoparticles with stable casein and polysaccharide through a one-step method.
A three-layer composite nanoparticle is formed by prolamin, sodium caseinate and polysaccharide, and is characterized in that the three-layer composite nanoparticle comprises an inner prolamin structure, a middle casein structure and an outer polysaccharide structure, wherein the middle casein structure and the outer polysaccharide structure jointly coat the inner prolamin structure to form a three-layer composite nanoparticle structure.
A preparation method of three-layer composite nano-particles formed by prolamin, sodium caseinate and polysaccharide is characterized in that the three-layer composite nano-particles are obtained by a one-step nano-precipitation method, and the preparation method specifically comprises the following steps:
(1) dissolving prolamin in organic phase, and stirring to completely dissolve;
(2) dissolving sodium caseinate and polysaccharide in water phase together, and stirring to dissolve completely;
(3) dropwise adding the alcohol soluble protein organic phase obtained in the step (1) into the water phase obtained in the step (2) under the stirring condition to obtain dispersion, and stirring by adopting a magnetic stirrer, wherein the stirring speed of the magnetic stirrer is 500-900 rpm.
(4) And (3) carrying out rotary evaporation on the dispersion liquid obtained in the step (3) to remove an organic phase, wherein the rotary evaporation vacuum degree is 0.07-0.1 MPa, the temperature is 45-55 ℃, centrifuging to remove large particles to obtain a nanoparticle dispersion liquid, the centrifuging speed is 1000-5000 rpm, the centrifuging time is 5-40 min, then carrying out vacuum freeze drying to obtain a nanoparticle solid, and the temperature of the vacuum freeze drying is preferably-60-37 ℃, and more preferably-40 ℃. The time of the vacuum freeze drying is preferably 15-28 h, and more preferably 24 h;
the alcohol soluble protein is plant-extracted or commercially-available conventional alcohol soluble protein, the concentration range of the alcohol soluble protein is 0.5% -5% (w/v), preferably 0.5% -4%, the alcohol soluble protein is dissolved by adopting an organic phase, the organic phase comprises an ethanol aqueous solution, a methanol aqueous solution, an isopropanol aqueous solution, an acetone aqueous solution or ethylene glycol, the volume fraction of the ethanol aqueous solution is 30% -90%, the volume fraction of the methanol aqueous solution is 45% -95%, the volume fraction of the isopropanol aqueous solution is 40% -85%, preferably, the volume fraction of the organic phase methanol aqueous solution is 60% -90% (w/v), the volume fraction of the ethanol aqueous solution is 65% -85% (w/v), the volume fraction of the isopropanol aqueous solution is 70% -80% (v/v), the sodium caseinate and the polysaccharide are dissolved in the water phase together according to a certain proportion, the ratio of organic phase to aqueous phase ranges from 1: 1.5-10, preferably, the volume of the organic phase and the aqueous phase is 1: 2-6 (v/v), wherein the polysaccharide type is negative polysaccharide, the concentration of sodium casein is 0.15% -10% (w/v), the concentration of polysaccharide is 0.001% -7% (w/v), the concentration of sodium casein is 0.15% -0.3% (w/v), and the concentration of polysaccharide is 0.001% -1% (w/v).
The nano-particles have a three-layer structure, namely, the prolamin structure at the inner layer is a hydrophobic core formed by prolamin, the sodium casein structure at the middle layer is an amphiphilic sodium casein monolayer, and the hydrophilic polysaccharide surface of the polysaccharide structure at the outer layer is attached to form a shell, the size of the nano-particles prepared by the method is changed along with the variety of the polysaccharide at the outer layer and is different from 50-600 nm, and the polydispersity is less than 0.3.
The alcohol-soluble protein is zein, millet alcohol-soluble protein, hordein and the like;
aiming at the problems of poor stability and low embedding rate of prolamin nanoparticles, one of the purposes of the patent is to prepare a stable nanoparticle for embedding a medicament, so that the stability targeting property and the bioavailability of the medicament are improved.
Aiming at the complex problem of preparing multilayer nano particles at present, the invention provides a one-step method for preparing nano particles with the structure, so that the complexity of the preparation process is greatly reduced.
The negative polysaccharide is polysaccharide with negative charge, such as chondroitin sulfate, sulfate radical modified curdlan, fucoidin, sodium alginate, sodium carboxymethylcellulose and the like; the invention utilizes the self-assembly property of alcohol soluble protein, utilizes the acting force between biomacromolecules and adopts an anti-solvent method to prepare the nano-particles with hydrophobic alcohol soluble protein cores, casein sodium single layers and hydrophilic shells. The method has the advantages that a novel and simple method for preparing the nano particles with the multilayer composite structure is provided, the size distribution of the particles formed at the same time is uniform, the embedding rate of lipid solubility is obviously improved, and the stability and the biological activity of the nano particles are enhanced. The three-layer stable nanoparticle provided by the invention can be used for protecting and delivering drugs or functional factors.
The rotary evaporation, centrifuge and vacuum freeze-drying equipment are not particularly limited in the invention, and the equipment can be commercially available according to the conventional method in the field.
Example 1
1 g of prolamin was dissolved in 100 mL of 75% (w/v) aqueous ethanol, stirred to complete dissolution, and then filtered to remove insoluble material. The sodium caseinate and chondroitin sulfate were dissolved in water at concentrations of 4 mg/mL and 3 mg/mL, respectively. Then, 100 mL of ethanol aqueous solution of prolamin was added dropwise to 500 mL of aqueous solution containing sodium caseinate and chondroitin sulfate with stirring speed of 600 rpm, and stirring was continued for 15 min to obtain a dispersion. The ethanol is rotary evaporated at 40 ℃, and the precipitate is removed by centrifugation at 3000 rmp for 10 min to obtain the nano dispersion.
The shape characteristics of the obtained nanoparticles are observed by a scanning electron microscope and a transmission electron microscope, and the result shows that the prepared particles are in a small spherical shape with a regular surface and in a core-shell structure, namely, the casein sodium and the chondroitin sulfate are arranged on the outer layer of the prolamin particles.
Dynamic light scattering equipment showed that the average particle size of the nanoparticles was 200nm and the polydispersity was 0.15. Potentiometric analysis indicated that the charge of the particles as a function of pH was identical to that of chondroitin sulfate, which was thus present on the surface of sodium caseinate, and the results indicated that a hydrophobic prolamin core, an amphiphilic sodium caseinate monolayer and a hydrophilic chondroitin sulfate shell were formed.
The ionic strength and pH stability researches show that the prepared nanoparticles are stable in a pH range of 2-8 and can resist the ionic strength of 3M NaCl.
The infrared chromatography finds that the peptide bond between the sulfate radical on the chondroitin sulfate molecule and sodium caseinate forms a hydrogen bond and an ionic bond.
It was found that the nanoparticles still have the characteristics of nanoparticles by re-dissolving after freeze-drying the nanoparticles.
Example 2
2 g of prolamin was dissolved in 100 mL of 85% (w/v) aqueous isopropanol, stirred to complete dissolution, and then filtered to remove insoluble material. The concentration of sodium caseinate and hyaluronic acid dissolved in water is 2 mg/mL and 4 mg/mL respectively. Then, 100 mL of ethanol aqueous solution of prolamin was added dropwise to 300 mL of aqueous solution containing sodium caseinate and hyaluronic acid with stirring speed of 600 rpm, and stirring was continued for 30 min to obtain a dispersion. The ethanol is evaporated by rotation at the temperature of 45 ℃, and the nano dispersion liquid is obtained by centrifuging for 10 min at the temperature of 4000 rmp.
The shape characteristics of the obtained nanoparticles are observed by a scanning electron microscope, and the result shows that the prepared particles are in a small spherical shape with a regular surface and in a core-shell structure, namely, the casein sodium and the hyaluronic acid are arranged on the outer layer of the prolamin particles.
The morphology characteristics of the obtained nanoparticles are observed by a scanning electron microscope and a transmission electron microscope, and the results show that:
dynamic light scattering showed an average nanoparticle size of 230nm and a polydispersity of 0.17. The potentiometric analysis indicated that the charge of the particles as a function of pH was consistent with hyaluronic acid, which was thus present on the surface of sodium caseinate, and the results indicated that a hydrophobic prolamin core, an amphiphilic sodium caseinate monolayer and a hydrophilic hyaluronic acid shell were formed.
The ionic strength and pH stability researches show that the prepared nanoparticles are stable in a pH range of 4-8 and can resist the ionic strength of 2M NaCl.
The infrared chromatography finds that the peptide bond between the carboxylate radical on the hyaluronic acid molecule and sodium caseinate forms a hydrogen bond and an ionic bond.
It was found that the nanoparticles still have the characteristics of nanoparticles by re-dissolving after freeze-drying the nanoparticles.
Example 3
1 g of prolamin was dissolved in 100 mL of 90% (w/v) aqueous methanol, stirred to complete dissolution, and then filtered to remove insoluble material. The concentrations of sodium caseinate and fucose dissolved in water were 6 mg/mL and 2 mg/mL, respectively. Then, 100 mL of an aqueous ethanol solution of prolamin was added dropwise to 600 mL of an aqueous solution containing sodium caseinate and fucose while stirring at 800 rpm, and stirring was continued for 20 min to obtain a dispersion. The ethanol is rotary evaporated at 60 ℃, and the precipitate is removed by centrifugation at 3500 rmp for 8 min to obtain the nano dispersion liquid.
The shape characteristics of the obtained nanoparticles are observed by a scanning electron microscope, and the result shows that the prepared particles are in a small spherical shape with a regular surface and in a core-shell structure, namely, the casein sodium and the fucose are arranged on the outer layer of the prolamin particles.
Dynamic light scattering showed an average particle size of 147nm and a polydispersity of 0.21. Potentiometric analysis indicated that the charge of the particles as a function of pH was consistent with fucose and therefore fucose was present on the surface of sodium caseinate, resulting in the formation of a hydrophobic prolamin core, an amphiphilic sodium caseinate monolayer and a hydrophilic fucose shell.
The ionic strength and pH stability researches show that the prepared nanoparticles are stable in a pH range of 2-8 and can resist the ionic strength of 3.5M NaCl.
Infrared chromatographic analysis shows that the peptide bond between the sulfate radical on the fucose molecule and sodium casein forms hydrogen bond and ion bond.
It was found that the nanoparticles still have the characteristics of nanoparticles by re-dissolving after freeze-drying the nanoparticles.
Example 4
3 g of prolamin was dissolved in 100 mL of ethylene glycol solution, stirred to complete dissolution, and then filtered to remove insoluble material. The sodium caseinate and sodium carboxymethylcellulose were dissolved in water at concentrations of 10 mg/mL and 4 mg/mL, respectively. Then, 100 mL of ethanol aqueous solution of prolamin was added dropwise to 1000 mL of aqueous solution containing sodium caseinate and sodium carboxymethylcellulose at a stirring speed of 900 rpm, and stirring was continued for 40 min to obtain a dispersion. The ethanol is rotary evaporated at 50 ℃, and the precipitate is removed by centrifugation at 3000 rmp for 12 min to obtain the nano dispersion.
The morphology characteristics of the obtained nanoparticles are observed by a scanning electron microscope, and the result shows that the prepared particles are in a small spherical shape with a regular surface and have a core-shell structure, namely, the sodium caseinate and the sodium carboxymethylcellulose are arranged on the outer layer of the prolamin particles.
Dynamic light scattering showed an average particle size of 177nm and a polydispersity of 0.27. Potentiometric analysis indicated that the charge of the particles as a function of pH was consistent with chondroitin sulfate, so sodium carboxymethylcellulose was present on the surface of the sodium casein, and the results indicated that a hydrophobic prolamin core, an amphiphilic sodium casein monolayer, and a hydrophilic sodium carboxymethylcellulose shell were formed.
The ionic strength and pH stability researches show that the prepared nanoparticles are stable in a pH range of 2-8 and can resist the ionic strength of 3.5M NaCl.
Infrared chromatographic analysis shows that peptide bonds between carboxylate radicals on sodium carboxymethyl cellulose molecules and sodium casein form hydrogen bonds and ionic bonds.
It was found that the nanoparticles still have the characteristics of nanoparticles by re-dissolving after freeze-drying the nanoparticles.
Example 5
1.5 g of prolamin was dissolved in 100 mL of 85 aqueous ethanol, stirred to complete dissolution, and then filtered to remove insoluble material. The sodium caseinate and sulfate modified curdlan were dissolved in water at concentrations of 1 mg/mL and 2 mg/mL, respectively. Then 100 mL of ethanol aqueous solution of prolamin was added drop-wise to 800 mL of aqueous solution containing sodium caseinate and sulfated curdlan at 900 rpm stirring speed and stirring was continued for 40 min to obtain a dispersion. The ethanol is rotary evaporated at 50 ℃, and the precipitate is removed by centrifugation at 3000 rmp for 12 min to obtain the nano dispersion.
The morphology characteristics of the obtained nanoparticles are observed by a scanning electron microscope, and the result shows that the prepared particles are in a small spherical shape with a regular surface and in a core-shell structure, namely the casein sodium and the gel polysaccharide are arranged on the outer layer of the prolamin particles.
The morphology characteristics of the obtained nanoparticles are observed by a scanning electron microscope and a transmission electron microscope, and the results show that:
dynamic light scattering showed an average nanoparticle size of 159nm and a polydispersity of 0.27. The potentiometric analysis indicated that the charge of the particles as a function of pH was identical to that of chondroitin sulphate, which was therefore present on the surface of the sodium caseinate, and the results indicated that a hydrophobic prolamin core, an amphiphilic sodium caseinate monolayer and a hydrophilic gel polysaccharide shell were formed.
Through the ionic strength and pH temperature study, the prepared nanoparticles are stable in the pH range of 2-8 and can resist the ionic strength of 2.5M NaCl.
Infrared chromatographic analysis shows that the peptide bond between sulfate radical on sulfated curdlan molecule and sodium caseinate forms hydrogen bond and ion bond.
It was found that the nanoparticles still have the characteristics of nanoparticles by re-dissolving after freeze-drying the nanoparticles.
The above description is only a basic description of the concept of the present invention, and any modifications and embellishments made according to the technical solution of the present invention belong to the protection scope of the present invention.
Claims (8)
1. A three-layer composite nanoparticle is formed by prolamin, sodium caseinate and polysaccharide, and is characterized in that the three-layer composite nanoparticle comprises an inner prolamin structure, a middle casein structure and an outer polysaccharide structure, wherein the middle casein structure and the outer polysaccharide structure jointly coat the inner prolamin structure to form a three-layer composite nanoparticle structure.
2. A preparation method of three-layer composite nano-particles formed by prolamin, sodium caseinate and polysaccharide is characterized in that the three-layer composite nano-particles are obtained by a one-step nano-precipitation method, and the preparation method specifically comprises the following steps:
(1) dissolving prolamin in organic phase, and stirring to completely dissolve;
(2) dissolving sodium caseinate and polysaccharide in water phase together, and stirring to dissolve completely;
(3) dropwise adding the prolamin organic phase obtained in the step (1) into the water phase obtained in the step (2) under the stirring condition to obtain dispersion liquid.
(4) And (4) carrying out rotary evaporation on the dispersion liquid obtained in the step (3) to remove an organic phase, centrifuging to remove large particles to obtain a nano particle dispersion liquid, and then carrying out vacuum freeze drying to obtain a nano particle solid.
3. The method of claim 2, wherein the prolamin is a plant-derived or commercially-available prolamin having a prolamin concentration in the range of 0.5% to 5% (w/v).
4. The method of claim 2, wherein the prolamin is solubilized using an organic phase comprising an aqueous ethanol solution, an aqueous methanol solution, an aqueous isopropanol solution, an aqueous acetone solution, or ethylene glycol.
5. The method of claim 4, wherein the ethanol aqueous solution comprises 30% to 90% by volume, the methanol aqueous solution comprises 45% to 95% by volume, and the isopropyl alcohol aqueous solution comprises 40% to 85% by volume.
6. The method of claim 2, wherein the sodium caseinate and the polysaccharide are co-dissolved in the aqueous phase at a ratio of organic to aqueous phases in the range of 1: 1.5 to 10.
7. The method of claim 2, wherein the polysaccharide is a negative polysaccharide.
8. The method of claim 2, wherein the concentration of sodium caseinate ranges from 0.15% to 10% (w/v) and the concentration of polysaccharide ranges from 0.001% to 7% (w/v).
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| CN114835924A (en) * | 2022-06-20 | 2022-08-02 | 威海惠安康生物科技有限公司 | Preparation method of prolamin nanoparticles formed by co-assembling sodium caseinate and polysaccharide |
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| ES2697301A1 (en) * | 2017-07-21 | 2019-01-22 | Univ Del Pais Vasco / Euskal Herriko Unibertsitatea | NANOEMULSIONS TO PREVENT THE BIOLOGICAL COLONIZATION OF SURFACES (Machine-translation by Google Translate, not legally binding) |
| CN112715939A (en) * | 2020-12-30 | 2021-04-30 | 杭州娃哈哈科技有限公司 | Preparation method of dihydromyricetin compound solution capable of being stably stored for long time |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| ES2697301A1 (en) * | 2017-07-21 | 2019-01-22 | Univ Del Pais Vasco / Euskal Herriko Unibertsitatea | NANOEMULSIONS TO PREVENT THE BIOLOGICAL COLONIZATION OF SURFACES (Machine-translation by Google Translate, not legally binding) |
| CN112715939A (en) * | 2020-12-30 | 2021-04-30 | 杭州娃哈哈科技有限公司 | Preparation method of dihydromyricetin compound solution capable of being stably stored for long time |
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| CN114835924A (en) * | 2022-06-20 | 2022-08-02 | 威海惠安康生物科技有限公司 | Preparation method of prolamin nanoparticles formed by co-assembling sodium caseinate and polysaccharide |
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