CN113730373A - zein-AOS composite nano-particles for delivering curcumin and preparation method thereof - Google Patents
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Abstract
The invention provides zein-AOS composite nanoparticles for delivering curcumin and a preparation method thereof. Under the conditions of proper pH and temperature, the zein-AOS composite nano-particle prepared by the invention is relatively stable, the encapsulation rate of curcumin reaches 89.7%, the nano-particle can reduce the release (< 33%) of curcumin in simulated gastric juice (SGF, pH 2.0), and can fully release curcumin in simulated intestinal juice (SIF, pH 7.4), and the release rate reaches 83.8%.
Description
Technical Field
The invention relates to curcumin delivery nanoparticles, in particular to zein-AOS composite nanoparticles for delivering curcumin and a preparation method thereof.
Background
Curcumin (Cur) is present in Curcuma aromatica, and has antioxidant, antiinflammatory, antiulcer, antibacterial, and antiviral effects. In addition, curcumin also has good anticancer effect and has no toxicity to normal cells. However, its use is limited due to its poor physicochemical stability and low oral bioavailability.
In the past decades, in order to overcome the problems of poor stability and low oral bioavailability, a number of methods have been investigated including the preparation of liposome structures or phospholipid structures, etc., e.g. WO2007101551, which prepares a complex of phospholipids and curcumin, however, the resulting product is a sticky wax which renders the product essentially non-encapsulated, and many groups are exploring different ways to combine curcumin with auxiliary agents such as piperine, quercetin, etc.
Despite the advances in the art, there is still a great need to improve curcumin encapsulation efficiency, bioavailability and stability.
Disclosure of Invention
Through years of efforts, the research team utilizes a Fourier infrared spectrometer, a transmission electron microscope, a scanning electron microscope and a differential scanning calorimeter to characterize various composite nanoparticles, examines the stability of different composite nanoparticles under different ionic strength, pH value and temperature conditions, and researches the encapsulation and release performance of the nanoparticles, and unexpectedly discovers the zein-AOS composite nanoparticle which is stable, high in encapsulation efficiency and suitable for delivering curcumin.
One of the objectives of the present invention is to provide zein-AOS composite nanoparticles for delivering curcumin, which are composed of zein (zein) and alginate-derived oligosaccharides (AOS), wherein the weight ratio of the zein to the alginate-derived oligosaccharides is 2: 1.
Another object of the present invention is to provide a method for preparing zein-AOS composite nanoparticles for delivering curcumin, which comprises the steps of:
1) preparation of zein stock solution
0.4g of zein is dissolved in 20mL of 75% ethanol water solution, and then 10mL of zein solution is slowly dropped into 30mL of deionized water. The ethanol was removed from the sample at 40 ℃ using a rotary evaporator. Finally, the solution was made up to 40mL with deionized water and stored in a refrigerator at 4 ℃ until use.
2) Preparation of 0.2% brown algae oligosaccharide stock solution
Dissolving a certain amount of brown alginate oligosaccharide into deionized water to obtain 0.2% brown alginate oligosaccharide stock solution.
3) zein-AOS (2:1) preparation of nanoparticles
Diluting 2.5mL of brown alginate oligosaccharide stock solution to 18mL by using deionized water, slowly dripping 2mL of zein stock solution into 18mL of brown alginate oligosaccharide aqueous solution, and magnetically stirring for 1 h. And then adjusting the pH value of the zein-AOS composite nano system to 4. Centrifugation was carried out at 3000rpm for 10 minutes, the supernatant was preserved, and finally the sample was stored at 4 ℃ until use.
In another aspect, the invention provides the use of zein-AOS composite nanoparticles for the delivery of curcumin. The use of the zein-AOS composite nanoparticle in the delivery of curcumin comprises loading curcumin into the zein-AOS composite nanoparticle to form a Cur/zein-AOS composite nanoparticle.
Preferably, the weight ratio of Cur to zein to AOS in the Cur/zein-AOS composite nano particles is 1:100:50-4:100:50
The zein-AOS composite nano-particles can be further used as carriers of fat-soluble active substances.
The invention successfully prepares the zein-AOS composite nano-particles through electrostatic interaction, hydrogen bond interaction and hydrophobic interaction. Meanwhile, the invention researches that the proportion of zein and AOS has influence on the particle size, potential and PDI of the composite nano-particles, when the weight ratio of zein to AOS is 2:1, the prepared zein-AOS composite nano-particles are uniform and stable, and the composite nano-particles have good stability on ionic strength (0-12.5) mmol/L, temperature (30-90 ℃), pH (4-9) and the like.
The Cur/zein-AOS has gastrointestinal tract slow release performance, and the zein-AOS nano-particles prepared by the method can be used as ideal carriers of fat-soluble active substances.
Drawings
FIG. 1 shows particle size and PDI (a) and zeta (b) potentials of zein-AOS nanoparticles at different zein and AOS ratios.
FIG. 2 is an infrared spectrum of different nanoparticles, where a is AOS nanoparticles, b is zein nanoparticles, c is zein-AOS nanoparticles, d is Cur nanoparticles, and e is Cur/zein-AOS (e) nanoparticles.
FIG. 3 is SEM and TEM images of nanoparticles, where a is an SEM image of zein nanoparticles, b is an SEM image of zein-AOS (2:1) composite nanoparticles (b), c is a TEM image of zein nanoparticles, and d is a TEM image of zein-AOS composite nanoparticles.
Fig. 4 is a DSC profile of nanoparticles.
FIG. 5 shows the particle size, PDI, potential of zein-AOS nanoparticles at different ionic strengths (a, b), different pH conditions (c, d), and different temperatures (e, f).
FIG. 6 shows the encapsulation efficiency of Cur/zei-AOS nanoparticles when loaded with different Cur.
FIG. 7 is a graph of Cur, Cur/zein, Cur/zein-AOS which mimics the rate of Cur release in the gastrointestinal tract.
Detailed Description
The zein (zein) used in the invention is purchased from Sigma reagent company, Missouri, USA, and the alginate oligosaccharide AOS is purchased from Shandong Crystal group, Inc.
The particle size, Polydispersity (PDI) and zeta potential of the nanoparticles are tested by a Malvern potentiostat. Before the measurement, the sample was diluted with deionized water having a pH of 4 to an appropriate concentration (a counting rate of 100-.
The invention adopts a Fourier transform spectrophotometer at the wavelength of 400-4000cm-1Resolution of 4cm-1Characterizing Cur, Cur/zein, zein-AOS and Cur/zein-AOS.
The invention adopts a Differential Scanning Calorimeter (DSC) to analyze the thermal deformation characteristics of Cur, Cur/zein, zein-AOS and Cur/zein-AOS. Approximately 5mg of the lyophilized sample was placed in a sealed standard aluminum pan for testing. The nitrogen flow rate was set at 10 deg.C/min and the temperature was increased from 30 deg.C to 120 deg.C at a rate of 5 deg.C/min while an empty pan was used as a control.
According to the invention, a Transmission Electron Microscope (TEM) is used for characterizing the zein-AOS sample under 200kV accelerating voltage. The surface morphology of the zein-AOS was characterized by a Scanning Electron Microscope (SEM) at an accelerating voltage of 5.0 kV. Before characterization, zein-AOS liquid samples were dropped on silicon wafers, and the samples were coated with gold for observation after drying in air.
The present invention to evaluate the release of Cur/zein-AOS nanoparticles in the gastrointestinal tract, we prepared Simulated Gastric Fluid (SGF) at pH 2.0 using 3.2 mg/mL aqueous pepsin. The Simulated Intestinal Fluid (SIF) has a content of 6.8mg/mL K2HPO4Pancreatin 0.8mg/mL, bile extract 20mg/mL, NaCl 8.775mg/mL, the pH was adjusted to 7.4. 30mL of freshly prepared Cur/zein-AOS dispersion were mixed with 30mL of SGF and shaken at 100rpm for 90 minutes at 37 ℃. 2mL samples were taken at regular intervals (30min, 60min, 90min) and supplemented with 2mL fresh SGF. After 90min, the entire SGF digest was poured into 60mL SIF. 2mL samples were collected at 120,150,180,210 and 240 minutes. The fresh SIF solution was replenished with 2 mL. The collected solution was centrifuged at 10000 Xg for 5min to remove insoluble matter. Curcumin in the supernatant was measured at 426nm with a UV-2600 spectrophotometer.
The method for testing the encapsulation efficiency of curcumin in the nanoparticles comprises the following steps: the freeze-dried Cur/zein-AOS nanoparticles were dissolved with ultrasound-assisted in 80% aqueous ethanol (10 mL). After centrifugation at 4000rpm for 10min, the supernatant was collected. Washing the precipitate with 80% ethanol water solution for 3 times, centrifuging, and collecting supernatant. EE is determined according to the ratio of the curcumin content in the clear liquid to the total curcumin content. The curcumin content in the supernatant was determined by UV-2600 spectrophotometer at 426 nm.
Preparation of zein stock solution of 5mg/mL
0.4g of zein is dissolved in 20mL of 75% ethanol water solution, and then 10mL of zein solution is slowly dropped into 30mL of deionized water. The ethanol was removed from the sample at 40 ℃ using a rotary evaporator. Finally, the solution was made up to 40mL with deionized water and stored in a refrigerator at 4 ℃ until use.
Preparation of 2mg/mL brown algae oligosaccharide stock solution
2g of brown alginate oligosaccharide is dissolved in 1L of deionized water to obtain 2mg/mL brown alginate oligosaccharide stock solution.
Preparation of different curcumin stock solutions
Dissolving curcumin in anhydrous ethanol to obtain stock solutions of 1mg/mL,2mg/mL,3mg/mL and 4 mg/mL.
Preparation of Cur/zein stock solution
1mL of curcumin stock solution (1mg/mL,2mg/mL,3mg/mL,4mg/mL) was slowly dropped into 5mL of zein stock solution, respectively, and magnetically stirred (500rpm) for 2 hours. Then, the mixture solution was added dropwise to 15mL of deionized water, and magnetically stirred (500rpm) for 2 hours in a dark environment. Ethanol was removed by rotary evaporation. Finally, the sample was made up to 20mL with deionized water and stored in a 4 ℃ freezer for use.
Preparation example 1 zein nanoparticle preparation
0.4g of zein is dissolved in 20mL of 75% ethanol water solution, and then 10mL of zein solution is slowly dropped into 30mL of deionized water. The ethanol was removed from the sample at 40 ℃ using a rotary evaporator. Finally, the solution was made up to 40mL with deionized water and stored in a refrigerator at 4 ℃ until use.
Preparation example 2 preparation of zein-AOS nanoparticles with different compounding ratios
Diluting a certain amount of brown alginate oligosaccharide stock solution to 18mL by using deionized water, slowly dripping 2mL of zein stock solution into 18mL of brown alginate oligosaccharide aqueous solution to enable the dosage ratio of zein to brown alginate oligosaccharide to be 1:1,1:2,2:1,4:1,5:1,10:1 and 20:1 respectively, and magnetically stirring for 1 h. The pH of the zein-AOS composite nanoparticles was then adjusted to 4. Centrifugation was carried out at 3000rpm for 10 minutes, the supernatant was preserved, and finally the sample was stored at 4 ℃ until use.
Preparation example 3 preparation of Cur/zein-AOS nanoparticles with different ratios
The prepared Cur/zein stock solutions (2mL) added with curcumin stock solutions (1mg/mL,2mg/mL,3mg/mL and 4mg/mL) with different concentrations are respectively and slowly added into 18mL of alginate oligosaccharide aqueous solution (containing 2.5mL of alginate oligosaccharide stock solutions), the mixture is magnetically stirred for 1h, and then the pH value of the Cur/zein-AOS compound nanoparticles is respectively adjusted to 4. Finally the samples were stored at 4 ℃ for further studies.
EXAMPLE 1 particle size, zeta potential and PDI determination of different compounding ratios of zein-AOS nanoparticles
The particle size, zeta potential and PDI measurements of zein-AOS nanoparticles of different ratios in preparation example 4 were performed, and the results are shown in FIG. 1 that the particle size of zein (90nm) is smaller than that of zein-AOS. Since the negatively charged alginate oligosaccharides are not sufficient to stabilize zein nanoparticles, precipitation and flocculation can be observed when the zein-AOS weight ratio is 20: 1. As the amount of brown algae oligosaccharides increased (the ratio of zein to brown algae oligosaccharides increased from 10:1 to 2:1), the particle size of zein decreased from 128.7nm to 116.5 nm. This change may be due to a change in the electrostatic interaction between zein and AOS. The particle size of the zein-AOS increased from 116.5nm to 209.9nm as the ratio of zein to AOS increased from 2:1 to 1: 2. Probably because the zein nanoparticles are covered by multiple layers of alginate oligosaccharides. As the amount of brown algae oligosaccharide increased, the charge of the zein-AOS nanoparticle decreased (the ratio of zein to brown algae oligosaccharide decreased from 10:1 to 1:2, and the charge decreased from-23.2 mV to-32 mV). It is shown that the charge of the zein-AOS composite nanoparticle is mainly controlled by the negatively charged alginate oligosaccharide. When the weight ratio of zein-AOS is 2:1, the brown alginate oligosaccharide with negative charges can effectively stabilize the nanoparticles, and the particle size of the nanoparticles is small and uniform (the particle size is 116.5nm, and the PDI is 0.16). Therefore, we chose a 2:1 ratio for further study.
Example 2 FTIR Studies of different nanoparticles
The nanoparticles of zein, AOS, Cur, zein-AOS (2:1), Cur/zein-AOS (2:100:50) in the above preparation examples were characterized by infrared spectroscopy, and the results are shown in FIG. 2 at 3500 cm--1The band of (3) is O-H stretching vibration, and O-H stretching vibration of zein-AOS and zein/Cur-AOS is different from that of zein, AOS and Cur, and probably because hydrogen bonds are formed among the zein, Cur and AOS. In the spectrum of FIG. 2b, zein has a characteristic peak of 1655cm-1And 1538cm-1And represent amide I (C ═ O stretching vibration) and amide II (N ═ H bending vibration), respectively. After addition of Cur and AOS, amide I in the spectrum of zer-AOS and Cur/zein-AOS shifted to 1646cm-1And 1651cm-1Amide II moved to 1530cm respectively-1And 1535cm-1Indicating that there is an electrostatic interaction between zein, AOS and Cur. 1035cm in AOS-1The characteristic peak of (b) in the zein-AOS shifted to 1030cm-1The zein/Cur-AOS composite particles are moved to 1038cm-1To (3). As in the spectrum of FIG. 2d, 1423cm-1、1277cm-1、1162cm-1The peaks at (A) represent stretching vibration of an aromatic ring, part of "ketone", and stretching vibration of an inter-ring chain, respectively. However, these characteristic peaks disappeared in the spectrum of zein/Cur-AOS, indicating that there are hydrogen bonds and hydrophobic interactions between zein, curcumin and alginate oligosaccharides, and that the nanoparticles effectively encapsulate curcumin.
Example 3 SEM and TEM Studies of different nanoparticles
The morphology of zein and zein-AOS (2:1) in the preparation examples was characterized by scanning electron microscopy and transmission electron microscopy. As shown in fig. 3: the zein nanoparticles are spherical in structure, but the surfaces are not smooth, and slight adhesion exists among the particles. The phenomenon is that the zein has certain film forming property due to the intermolecular disulfide bond of the sulfur-containing amino acid, so that the microspheres have certain adhesion with the evaporation of the solvent. When the zein is combined with the alginate oligosaccharides, the shape of the nano particles is spherical, and the nano particles are not adhered. zein-AOS nanoparticles are smoother and larger in particle size than zein. This is due to the fact that brown algae oligosaccharides coat zein through electrostatic interaction and hydrogen bonding. TEM also showed that the zein-AOS nanoparticles were larger than the zein particle size.
Example 4 DSC investigation of different nanoparticles
The thermal denaturation of zein, Cur/zein (2:1), and Cur/zein-AOS (2:100:50) in the preparation examples was measured by Differential Scanning Calorimetry (DSC). As shown in FIG. 4, the decomposition temperature of zein was 79.8 ℃. The decomposition temperature (88.2 ℃) of the Cur/zein nanoparticles is higher than that of zein. This is probably because the Cur/zein nanoparticles form a new encapsulating structure. When Cur/zein is combined with AOS, a higher decomposition temperature (91.2 ℃) is detected, which indicates that the molecular interaction between the brown alginate oligosaccharide and the zein increases the decomposition temperature. The characteristic decomposition peak (179.2 ℃) of curcumin in the Cur/zein and Cur/zein-AOS nanoparticles disappeared, indicating that curcumin was well dispersed in nanoparticles in an amorphous form.
Example 5 stability Studies under different environmental conditions stability studies of different salt ionic strengths
At pH 4, different concentrations of NaCl (0,5,10,12.5,15,20 mM) were prepared. And mixing the zein-AOS dispersion liquid with NaCl solutions with the same volume and different concentrations to obtain a series of samples. The particle size and potential of the sample were measured by a malvern potentiometer.
Drug-loaded nanoparticles present various environmental challenges in the human gastrointestinal tract. Therefore, it is important to study the physical stability of the composite nanoparticles under different ionic strengths. As shown in FIGS. 5a and b, the particle size of the zein-AOS (2:1) complex was 116.5nm at a NaCl concentration of 0 mmol/L. When the ion concentration was increased to 10 mmol/L, the composite nanoparticle had a particle size of 278nm and PDI of 0.21, at which time the composite nanoparticle was relatively stable. However, as the ionic strength increases, the particle size and PDI of the composite nanoparticle increase, aggregation and precipitation occur in the range of 12.5 to 20mmol/L, and the amount of charge of the composite nanoparticle decreases. This is probably because the electrostatic shielding effect reduces the surface charge of zein with increasing ionic strength, thereby weakening the interaction between zein and alginate oligosaccharides.
Influence of the pH of the solution
The pH of the zein-AOS dispersion (pH 2 to pH 8) was adjusted with 1M HCl or NaOH. The particle size and zeta potential were then obtained using a malvern potentiometer.
The composite nanoparticles as carriers of curcumin will be subject to pH changes in the gastrointestinal tract. Therefore, it is important to study the pH stability of the composite nanoparticle. As shown in fig. 5c, d: at pH 3, the zein-AOS nanoparticles were unstable (particle size 1424.3nm, PDI 0.47) and aggregated, probably due to the reduced surface charge and low absolute potential of the nanoparticles, and the reduced electrostatic repulsion between the nanoparticles. As the pH value increases (pH 4-pH 9), the absolute potential of the zein-AOS particles increases, the particle size is less than 135nm, and the PDI is less than 0.23. When the pH value is 4-9, the electrostatic interaction force among the nano particles can keep the nano system stable. Therefore, zein-AOS composite nanoparticles can be used as ideal carriers.
Influence of temperature
And heating the zein-AOS dispersion liquid at 30-90 ℃ for 30min, cooling to room temperature, and measuring the particle size and the potential of the sample.
The effect of temperature on zein-AOS composite nanoparticles is shown in FIG. 5e, f: after heating for 30min at 30-90 ℃, the particle size (129.9nm-136.5nm), PDI (0.16-0.19) and charge (-29.5 mV-30.5 mV) of the composite nano-particles are not changed greatly, which indicates that the composite nano-particles have good temperature stability. This result can be explained by the thermal denaturation of zein at around 100 ℃.
Example 6 encapsulation efficiency (EE%) comparison study for different Cur loadings
The encapsulation efficiency of the zein-AOS (2; 1) composite nanoparticles is shown in FIG. 6: in this study, different amounts of curcumin (1mg,2mg,3mg,4mg) were loaded into zein-AOS complex nanoparticles. As curcumin increased, the EE of Cur/zein-AOS gradually decreased. Wherein, when curcumin was increased from 1mg to 4mg, the encapsulation efficiency decreased from 92.11% to 59.6%, indicating that excess curcumin exceeded the loading capacity of the zein-AOS complex nanoparticles. Therefore, in further studies, 2mg of curcumin was encapsulated in zein-AOS nanoparticles, where the ratio of curcumin, zein, and alginate oligosaccharides was 2:100: 50.
Example 7 comparative experiment for simulation of gastrointestinal environment digestion
The Cur, Cur/zein (2:25) and Cur/zein-AOS (2:100:50) nanoparticles in the preparation examples were used to simulate the release rate in the gastrointestinal tract, and as shown in FIG. 7, the curcumin release rates in Cur, Cur/zein and Cur/zein-AOS in Simulated Gastric Fluid (SGF) were 60.5%, 39.9% and 32.9%, respectively. The release rate of the free curcumin is obviously higher than that of Cur/zein and Cur/zein-AOS, and the zein has the effect of controlling the release of the curcumine.
In Simulated Intestinal Fluid (SIF), both Cur/zein and Cur/zein-AOS showed burst effect, probably because bile salts and polypeptides in SIF helped to dissolve curcumin, and in addition, AOS coated on zein surface were also dissolved. The release rate of Cur/zein is higher than that of Cur/zein-AOS. This is probably because the brown algae oligosaccharide layer can protect zein to some extent and can prevent zein from being rapidly hydrolyzed. Cur/zein-AOS has the effect of controlling and releasing curcumin in the stomach, and can release a large amount of curcumin in intestinal juice, so zein-AOS nanoparticles can be used as an ideal carrier of fat-soluble medicines.
Claims (6)
1. A zein-AOS composite nanoparticle for delivering curcumin is characterized in that the zein-AOS composite nanoparticle is composed of zein and Alginate Oligosaccharides (AOS), and the weight ratio of the zein to the alginate oligosaccharides is 2: 1.
2. A method of preparing zein-AOS composite nanoparticles for delivering curcumin, characterized in that said method of preparing zein-AOS composite nanoparticles comprises the steps of:
1) preparation of zein stock solution
0.4g of zein is dissolved in 20mL of 75% ethanol water solution, and then 10mL of zein solution is slowly dropped into 30mL of deionized water. The ethanol was removed from the sample at 40 ℃ using a rotary evaporator. Finally, the solution was made up to 40mL with deionized water and stored in a refrigerator at 4 ℃ until use.
2) Preparation of 0.2% brown algae oligosaccharide stock solution
Dissolving a certain amount of brown alginate oligosaccharide into deionized water to obtain 0.2% brown alginate oligosaccharide stock solution.
3) zein-AOS (2:1) preparation of nanoparticles
Diluting 2.5mL of brown alginate oligosaccharide stock solution to 18mL by using deionized water, slowly dripping 2mL of zein stock solution into 18mL of brown alginate oligosaccharide aqueous solution, and magnetically stirring for 1 h. And then adjusting the pH value of the zein-AOS composite nano system to 4. Centrifugation was carried out at 3000rpm for 10 minutes, the supernatant was preserved, and finally the sample was stored at 4 ℃ until use.
3. Use of zein-AOS composite nanoparticles as claimed in claim 1 for the delivery of curcumin.
4. Use of zein-AOS composite nanoparticles according to claim 4 in the delivery of curcumin, characterized in that said step of delivering curcumin comprises loading curcumin into the zein-AOS composite nanoparticles to form Cur/zein-AOS composite nanoparticles.
5. Use of zein-AOS composite nanoparticles according to claim 5 in the delivery of curcumin, characterized in that the weight ratio of Cur: zein: AOS in said Cur/zein-AOS composite nanoparticles is from 1:100:50 to 4:100: 50.
6. Use of zein-AOS composite nanoparticles as defined in claim 1 as carriers for fat-soluble active substances.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070254848A1 (en) * | 2004-03-24 | 2007-11-01 | Meiyu Geng | Algin Oligosaccharides and the Derivatives thereof as Well as the Manufacture and the Use of the Same |
| CN104814929A (en) * | 2015-05-14 | 2015-08-05 | 中国计量学院 | Preparation method of algin oligosaccharide liposome |
| CN105456196A (en) * | 2015-11-23 | 2016-04-06 | 上海交通大学 | Curcumin composite particle and preparation method and application thereof |
| CN106822035A (en) * | 2017-03-31 | 2017-06-13 | 中国农业大学 | A kind of zeins shellac curcumin composite colloid particle and preparation method thereof |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070254848A1 (en) * | 2004-03-24 | 2007-11-01 | Meiyu Geng | Algin Oligosaccharides and the Derivatives thereof as Well as the Manufacture and the Use of the Same |
| CN104814929A (en) * | 2015-05-14 | 2015-08-05 | 中国计量学院 | Preparation method of algin oligosaccharide liposome |
| CN105456196A (en) * | 2015-11-23 | 2016-04-06 | 上海交通大学 | Curcumin composite particle and preparation method and application thereof |
| CN106822035A (en) * | 2017-03-31 | 2017-06-13 | 中国农业大学 | A kind of zeins shellac curcumin composite colloid particle and preparation method thereof |
Non-Patent Citations (4)
| Title |
|---|
| YONGKAI YUAN 等: "One-step self-assembly of curcumin-loaded zein/sophorolipid nanoparticles: physicochemical stability, redispersibility, solubility and bioaccessibility", 《FOOD & FUNCTION》 * |
| 董潇 等: "玉米醇溶蛋白-葡萄糖美拉德反应产物制备姜黄素纳米颗粒", 《中国食品学报》 * |
| 黄旭琳 等: "负载姜黄素的玉米醇溶蛋白-多糖纳米颗粒的制备及生物活性研究", 《广东药学院学报》 * |
| 黄晓霞 等: "负载姜黄素的玉米醇溶蛋白-果胶纳米颗粒制备及抗氧化活性研究", 《广东农业科学》 * |
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