CN114712327A - Preparation method of curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles - Google Patents

Preparation method of curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles Download PDF

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CN114712327A
CN114712327A CN202210478417.3A CN202210478417A CN114712327A CN 114712327 A CN114712327 A CN 114712327A CN 202210478417 A CN202210478417 A CN 202210478417A CN 114712327 A CN114712327 A CN 114712327A
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rice bran
curcumin
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bran protein
sodium caseinate
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王娜
孙妍
陈长兰
肖志刚
赵凤芹
谭莲兴
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Liaoning University
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Abstract

The invention relates to a preparation method of curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles, and belongs to the technical field of nanoparticle preparation. The preparation method comprises the following steps: extracting rice bran protein from defatted rice bran to prepare rice bran protein stock solution; mixing the rice bran protein stock solution, the sodium caseinate stock solution, the curcumin-ethanol solution and the Arabic gum stock solution under continuous stirring; removing ethanol by rotary evaporation, centrifuging, and freeze drying to obtain curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles. The preparation method is simple and low in cost, and the prepared nanoparticles have controllable particle size, higher stability and good biocompatibility, and can improve the solubility, dispersibility, stability and bioavailability of curcumin to a great extent.

Description

Preparation method of curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles
Technical Field
The invention belongs to the technical field of nanoparticle preparation, and particularly relates to a preparation method of curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles.
Background
Curcumin is a hydrophobic plant polyphenol, naturally occurring in the rhizomes of plants of the Zingiberaceae family. Curcumin has been shown to possess a variety of biological activities, including antibacterial, antidiabetic, antioxidant, anti-inflammatory, antitumor, antiviral, cardioprotective, neuroprotective, and the like. But curcumin is used as a highly hydrophobic molecule, the water solubility is only 11ng/mL, most curcumin can be discharged out of the body after being eaten by a human body, and the bioavailability is extremely low; curcumin is chemically unstable and susceptible to the polarity of the solution, surfactants, antioxidants, high temperature, light, extreme pH, moisture and oxygen. Curcumin is often limited in its utility as a nutritional ingredient in functional foods and beverages due to its chemical instability, low water solubility and low oral bioavailability. For this reason, researchers have attempted to embed curcumin in particles or emulsions with the goal of improving curcumin bioavailability.
Nanoparticles are typically in the size range of 10-1000nm, are readily dispersible and suspendable in water, and protein sources for the construction of protein nanoparticles include two categories: animal proteins and plant proteins. Plant protein has wider sources than animal protein and has a series of advantages of sustainability and the like, so the plant protein is more suitable to be used as a raw material for preparing nano particles in theory. However, when the pH of the single protein nanoparticle delivery system reaches the isoelectric point of the protein in the external environment, the protein structure is destroyed, so that a large amount of irreversible precipitation occurs, and the situation that the protein is degraded by the extreme pH and the complex enzyme system in the gastrointestinal tract environment cannot be solved. Therefore, it is necessary to develop a nanoparticle preparation method with higher stability and resistance by compounding proteins with other substances to construct nanoparticles, and nanoparticles for embedding, protecting and delivering drugs or nutritional ingredients are needed, and the nanoparticles prepared based on the method have better application prospects.
The invention patent (publication No. CN110917137B, publication No. 2021.08.10) discloses a preparation method of pickering emulsion with stable coordination of prolamin nanoparticles and starch nanoparticles, the pickering emulsion prepared by the invention has high stability, the embedding rate and the bioavailability of curcumin are improved, but the preparation period of the starch nanoparticles in the preparation process is long, and the preparation method of the pickering emulsion is complicated.
Disclosure of Invention
Aiming at the existing problems, the invention provides a method for preparing rice bran protein-Arabic gum-sodium caseinate nano-particles loaded with curcumin,
the technical scheme adopted by the invention is as follows:
a method for preparing curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles comprises the following steps:
1) mixing defatted rice bran with deionized water with the volume of 5 times, homogenizing the mixture for 3-5 min, stirring for 1-2 h at normal temperature, adjusting the pH of the mixture to 9.5 by using 3.0M sodium hydroxide, magnetically stirring for 1-2 h at 55 ℃, centrifuging for 30min at room temperature at 10000r/min, filtering the centrifuged supernatant with gauze, adjusting the pH of the filtrate to 4.4 by using 2.0M hydrochloric acid, standing for 30min, centrifuging for 30min at room temperature at 10000r/min, taking precipitate, washing the precipitate for 2 times by using deionized water to obtain a neutral protein solution, freezing for 24h at-20 ℃, then placing the neutral protein solution in a freeze dryer, and freeze-drying for 48h at-0.1-0.5 mbar, -60 ℃ to-50 ℃ to obtain rice bran protein;
2) dissolving the rice bran protein obtained in the step 1) in deionized water to prepare a 10mg/mL rice bran protein solution, and adjusting the pH of the solution by using 1.0M sodium hydroxide to obtain a rice bran protein stock solution;
3) at normal temperature, placing a NaCl solution on a magnetic stirrer with the rotation speed of 600r/min, adding the rice bran protein stock solution and the sodium caseinate stock solution obtained in the step 2) while stirring, then adjusting the pH of the mixed solution to 10.0-12.0 by using 1.0M sodium hydroxide, continuing to magnetically stir for 30min, adding a curcumin-ethanol solution under the dark condition, fully and uniformly mixing, adding an Arabic gum stock solution, continuing to stir for 1-2 h, rotationally evaporating to remove ethanol, slowly adjusting the pH of the mixed system by using 0.05M hydrochloric acid, finally centrifuging for 10min at 3000r/min, and taking a centrifuged supernatant to obtain a curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticle aqueous solution;
4) freezing the curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticle aqueous solution obtained in the step 3) at-20 ℃ for 24 hours, then placing the solution in a freeze dryer, and freeze-drying the solution at-60 ℃ to-50 ℃ for 48 hours at 0.1-0.5 mbar to obtain curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles.
Further, in the preparation method, in the step 2), the pH of the rice bran protein stock solution is 10.0-12.0.
Further, in the preparation method, in the step 3), the concentration of the NaCl solution is 5-25 mmol/L.
Further, in the above preparation method, step 3), the preparation method of the sodium caseinate stock solution comprises the following steps: sodium caseinate powder was dissolved in deionized water to give 10mg/mL sodium caseinate stock solution.
Further, in the above preparation method, step 3), the preparation method of the curcumin-ethanol solution comprises the following steps: dissolving curcumin powder in anhydrous ethanol to obtain 0.15mg/mL curcumin-ethanol solution, and storing at 4 deg.C in dark place.
Further, in the above preparation method, step 3), the preparation method of the acacia gum stock solution comprises: acacia gum powder was dissolved in deionized water to give a 10mg/mL acacia gum stock solution.
Further, in the preparation method, in the step 3), the NaCl solution, the rice bran protein stock solution, the sodium caseinate stock solution, the curcumin-ethanol solution, and the acacia gum stock solution are 15:1: 0.5-2.5: 18-21: 2 by volume ratio.
Further, in the preparation method, in the step 3), the pH of the mixed system is 4.0-4.8.
The particle size of the rice bran protein-Arabic gum-sodium caseinate nano-particles loaded with curcumin prepared by any one of the preparation methods is 310.2-1000 nm, the PDI is 0.136-0.491, the Zeta potential is-14.2-20.2 mV, the curcumin embedding rate is 58.3-98.5%, the bioavailability of curcumin in simulated gastric juice is 15.1-18.3%, and the bioavailability in simulated intestinal juice is 23.1-27.2%.
The invention has the beneficial effects that:
1. the preparation method provided by the invention is simple, the preparation period is short, the preparation cost is low, and the prepared rice bran protein-Arabic gum-sodium caseinate nanoparticles are excellent in performance, controllable in particle size, uniform in particle size and distribution, higher in stability and good in biocompatibility.
2. According to the invention, the rice bran protein-Arabic gum nano-particles are constructed by compounding protein and polysaccharide to improve the surface property of the nano-particles and the resistance to the external environment, and meanwhile, sodium caseinate is added into the rice bran protein-Arabic gum nano-particles as an emulsifier to improve the biological stability of the nano-particles and the embedding effect of curcumin, so that the stability, the dispersibility and the solubility of the curcumin are improved, and the bioavailability of the curcumin can be obviously improved.
3. The method has important significance for improving high-value utilization of rice bran which is a rice byproduct, reducing waste of resources and promoting application and development of rice bran protein, and the development of a rice bran protein-based nanoparticle delivery system also provides a new method and a new thought for developing a novel carrier material in the future.
Drawings
Fig. 1 is a graph showing the particle size and PDI of curcumin-loaded rice bran protein-gum arabic-sodium caseinate nanoparticles prepared in examples 1 to 5.
FIG. 2 is a Zeta potential diagram of curcumin-loaded rice bran protein-gum arabic-sodium caseinate nanoparticles prepared in examples 1-5.
Fig. 3 is a graph of the embedding rate of curcumin by the curcumin-loaded rice bran protein-gum arabic-sodium caseinate nanoparticles prepared in examples 1 to 5.
Fig. 4 is a scanning electron microscope image of curcumin-loaded rice bran protein-gum arabic-sodium caseinate nanoparticles prepared in example 3, wherein a is 20000 times amplified, and B is 40000 times amplified.
Fig. 5 is a graph showing the effect of curcumin bioavailability in curcumin-loaded rice bran protein-gum arabic-sodium caseinate nanoparticles prepared in example 3.
Detailed Description
Example 1
A method for preparing curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles comprises the following specific steps:
1) mixing 1kg of defatted rice bran with 5000mL of deionized water, homogenizing the mixture for 5min, stirring at normal temperature for 1h, adjusting the pH of the mixture to 9.5 by using 3.0M sodium hydroxide, magnetically stirring at 55 ℃ for 1.5h, centrifuging at room temperature for 30min at 10000r/min, removing insoluble substances, centrifuging supernatant, filtering with gauze, adjusting the pH of the filtrate to 4.4 by using 2.0M hydrochloric acid, standing for 30min, centrifuging at room temperature for 30min at 10000r/min, taking precipitate, washing the precipitate with deionized water for 2 times to obtain a neutral protein solution, freezing at-20 ℃ for 24h, then placing in a freeze dryer, and freeze-drying at-0.5 mbar and-55 ℃ for 48h to obtain rice bran protein;
2) dissolving 1g of the rice bran protein obtained in the step 1) in 100mL of deionized water to prepare a 10mg/mL rice bran protein solution, and adjusting the pH of the solution to 10.0 by using 1.0M sodium hydroxide to obtain a 10mg/mL rice bran protein stock solution;
3) dissolving 1g of Arabic gum powder in 100mL of deionized water to obtain Arabic gum stock solution of 10 mg/mL;
4) dissolving 1g of sodium caseinate powder in 100mL of deionized water to obtain 10mg/mL of sodium caseinate stock solution;
5) dissolving 15mg curcumin powder in 100mL absolute ethanol to obtain 0.15mg/mL curcumin-ethanol solution, and storing at 4 deg.C in dark place for use;
6) at normal temperature, 15mL of 10mmol/L NaCl solution is placed on a magnetic stirrer with the rotating speed of 600r/min, 1mL of the rice bran protein stock solution obtained in the step 2) and 1mL of the sodium caseinate stock solution obtained in the step 4) are added while stirring, then adjusting the pH value of the mixed solution to 12.0 by using 1.0M sodium hydroxide, continuing to magnetically stir for 30min, adding 19mL of curcumin-ethanol solution obtained in the step 5) under dark condition, fully and uniformly mixing, adding 2mL of acacia gum stock solution obtained in step 3), stirring for 1h, removing ethanol by using a rotary evaporator, slowly adjusting the pH of the mixed system to 4.0 by using 0.05M hydrochloric acid, finally centrifuging for 10min at 3000r/min, removing any insoluble large particles, taking centrifugal supernatant to obtain curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticle aqueous solution;
7) freezing the curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticle aqueous solution obtained in the step 6) at-20 ℃ for 24 hours, then placing the solution in a freeze dryer, and freeze-drying the solution at-0.5 mbar and-55 ℃ for 48 hours to obtain curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles.
Example 2
A method for preparing curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles comprises the following specific steps:
1) mixing 1kg of defatted rice bran with 5000mL of deionized water, homogenizing the mixture for 5min, stirring at normal temperature for 1h, adjusting the pH of the mixture to 9.5 by using 3.0M sodium hydroxide, magnetically stirring at 55 ℃ for 1.5h, centrifuging at room temperature for 30min at 10000r/min, removing insoluble substances, centrifuging supernatant, filtering with gauze, adjusting the pH of the filtrate to 4.4 by using 2.0M hydrochloric acid, standing for 30min, centrifuging at room temperature for 30min at 10000r/min, taking precipitate, washing the precipitate with deionized water for 2 times to obtain a neutral protein solution, freezing at-20 ℃ for 24h, then placing in a freeze dryer, and freeze-drying at-0.5 mbar and-55 ℃ for 48h to obtain rice bran protein;
2) dissolving 1g of the rice bran protein obtained in the step 1) in 100mL of deionized water to prepare a 10mg/mL rice bran protein solution, and adjusting the pH of the solution to 10.0 by using 1.0M sodium hydroxide to obtain a 10mg/mL rice bran protein stock solution;
3) dissolving 1g of Arabic gum powder in 100mL of deionized water to obtain Arabic gum stock solution of 10 mg/mL;
4) dissolving 1g of sodium caseinate powder in 100mL of deionized water to obtain 10mg/mL of sodium caseinate stock solution;
5) dissolving 15mg curcumin powder in 100mL absolute ethanol to obtain 0.15mg/mL curcumin-ethanol solution, and storing at 4 deg.C in dark place for use;
6) at normal temperature, 15mL of 10mmol/L NaCl solution is placed on a magnetic stirrer with the rotating speed of 600r/min, 1mL of the rice bran protein stock solution obtained in the step 2) and 1mL of the sodium caseinate stock solution obtained in the step 4) are added while stirring, then adjusting the pH value of the mixed solution to 12.0 by using 1.0M sodium hydroxide, continuing to magnetically stir for 30min, adding 20mL of curcumin-ethanol solution obtained in the step 5) under dark condition, fully and uniformly mixing, adding 2mL of acacia gum stock solution obtained in step 3), stirring for 1h, removing ethanol with a rotary evaporator, slowly adjusting pH of the mixed system to 4.2 with 0.05M hydrochloric acid, centrifuging at 3000r/min for 10min, removing any insoluble large particles, taking centrifugal supernatant to obtain curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticle aqueous solution;
7) freezing the curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticle aqueous solution obtained in the step 6) at-20 ℃ for 24 hours, then placing the solution in a freeze dryer, and freeze-drying the solution at-0.5 mbar and-55 ℃ for 48 hours to obtain curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles.
Example 3
A method for preparing curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles comprises the following specific steps:
1) mixing 1kg of defatted rice bran with 5000mL of deionized water, homogenizing the mixture for 5min, stirring at normal temperature for 1h, adjusting the pH of the mixture to 9.5 by using 3.0M sodium hydroxide, magnetically stirring at 55 ℃ for 1.5h, centrifuging at room temperature for 30min at 10000r/min, removing insoluble substances, centrifuging supernatant, filtering with gauze, adjusting the pH of the filtrate to 4.4 by using 2.0M hydrochloric acid, standing for 30min, centrifuging at room temperature for 30min at 10000r/min, taking precipitate, washing the precipitate with deionized water for 2 times to obtain a neutral protein solution, freezing at-20 ℃ for 24h, then placing in a freeze dryer, and freeze-drying at-0.5 mbar and-55 ℃ for 48h to obtain rice bran protein;
2) dissolving 1g of the rice bran protein obtained in the step 1) in 100mL of deionized water to prepare a 10mg/mL rice bran protein solution, and adjusting the pH of the solution to 10.0 by using 1.0M sodium hydroxide to obtain a 10mg/mL rice bran protein stock solution;
3) dissolving 1g of Arabic gum powder in 100mL of deionized water to obtain Arabic gum stock solution of 10 mg/mL;
4) dissolving 1g of sodium caseinate powder in 100mL of deionized water to obtain 10mg/mL of sodium caseinate stock solution;
5) dissolving 15mg curcumin powder in 100mL absolute ethanol to obtain 0.15mg/mL curcumin-ethanol solution, and storing at 4 deg.C in dark place for use;
6) at normal temperature, 15mL of 15mmol/L NaCl solution is placed on a magnetic stirrer with the rotating speed of 600r/min, 1mL of rice bran protein stock solution obtained in the step 2) and 1.5mL of sodium caseinate stock solution obtained in the step 4) are added while stirring, then adjusting the pH value of the mixed solution to 12.0 by using 1.0M sodium hydroxide, continuing to magnetically stir for 30min, adding 19mL of curcumin-ethanol solution obtained in the step 5) under dark condition, fully and uniformly mixing, adding 2mL of acacia gum stock solution obtained in step 3), stirring for 1h, removing ethanol by using a rotary evaporator, slowly adjusting the pH of the mixed system to 4.2 by using 0.05M hydrochloric acid, finally centrifuging for 10min at 3000r/min, removing any insoluble large particles, taking centrifugal supernatant to obtain curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticle aqueous solution;
7) freezing the curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticle aqueous solution obtained in the step 6) at-20 ℃ for 24 hours, then placing the frozen curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticle aqueous solution in a freeze dryer, and freeze-drying the frozen curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticle aqueous solution at-55 ℃ for 48 hours at 0.5mbar to obtain the curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticle.
Example 4
A method for preparing curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles comprises the following specific steps:
1) mixing 1kg of defatted rice bran with 5000mL of deionized water, homogenizing the mixture for 5min, stirring at normal temperature for 1h, adjusting the pH of the mixture to 9.5 by using 3.0M sodium hydroxide, magnetically stirring at 55 ℃ for 1.5h, centrifuging at room temperature for 30min at 10000r/min, removing insoluble substances, centrifuging supernatant, filtering with gauze, adjusting the pH of the filtrate to 4.4 by using 2.0M hydrochloric acid, standing for 30min, centrifuging at room temperature for 30min at 10000r/min, taking precipitate, washing the precipitate with deionized water for 2 times to obtain a neutral protein solution, freezing at-20 ℃ for 24h, then placing in a freeze dryer, and freeze-drying at-0.5 mbar and-55 ℃ for 48h to obtain rice bran protein;
2) dissolving 1g of the rice bran protein obtained in the step 1) in 100mL of deionized water to prepare a 10mg/mL rice bran protein solution, and adjusting the pH of the solution to 12.0 by using 1.0M sodium hydroxide to obtain a 10mg/mL rice bran protein stock solution;
3) dissolving 1g of Arabic gum powder in 100mL of deionized water to obtain Arabic gum stock solution of 10 mg/mL;
4) dissolving 1g of sodium caseinate powder in 100mL of deionized water to obtain 10mg/mL of sodium caseinate stock solution;
5) dissolving 15mg curcumin powder in 100mL absolute ethanol to obtain 0.15mg/mL curcumin-ethanol solution, and storing at 4 deg.C in dark place for use;
6) at normal temperature, 15mL of 20mmol/L NaCl solution is placed on a magnetic stirrer with the rotating speed of 600r/min, 1mL of rice bran protein stock solution obtained in the step 2) and 1.5mL of sodium caseinate stock solution obtained in the step 4) are added while stirring, then adjusting the pH value of the mixed solution to 12.0 by using 1.0M sodium hydroxide, continuing to magnetically stir for 30min, adding 20mL of curcumin-ethanol solution obtained in the step 5) under the dark condition, fully and uniformly mixing, adding 2mL of acacia gum stock solution obtained in step 3), stirring for 1h, removing ethanol with a rotary evaporator, slowly adjusting pH of the mixed system to 4.8 with 0.05M hydrochloric acid, centrifuging at 3000r/min for 10min, removing any insoluble large particles, taking centrifugal supernatant to obtain curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticle aqueous solution;
7) freezing the curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticle aqueous solution obtained in the step 6) at-20 ℃ for 24 hours, then placing the solution in a freeze dryer, and freeze-drying the solution at-0.5 mbar and-55 ℃ for 48 hours to obtain curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles.
Example 5
A method for preparing curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles comprises the following specific steps:
1) mixing 1kg of defatted rice bran with 5000mL of deionized water, homogenizing the mixture for 5min, stirring at normal temperature for 1h, adjusting the pH of the mixture to 9.5 by using 3.0M sodium hydroxide, magnetically stirring at 55 ℃ for 1.5h, centrifuging at room temperature for 30min at 10000r/min, removing insoluble substances, centrifuging supernatant for gauze filtration, adjusting the pH of the filtrate to 4.4 by using 2.0M hydrochloric acid, standing for 30min, centrifuging at room temperature for 30min at 10000r/min, taking precipitate, washing the precipitate with deionized water for 2 times to obtain a neutral protein solution, freezing at-20 ℃ for 24h, then placing in a freeze dryer, and freeze-drying at-0.5 mbar and-55 ℃ for 48h to obtain rice bran protein;
2) dissolving 1g of the rice bran protein obtained in the step 1) in 100mL of deionized water to prepare a 10mg/mL rice bran protein solution, and adjusting the pH of the solution to 10.0 by using 1.0M sodium hydroxide to obtain a 10mg/mL rice bran protein stock solution;
3) dissolving 1g of Arabic gum powder in 100mL of deionized water to obtain Arabic gum stock solution of 10 mg/mL;
4) dissolving 1g of sodium caseinate powder in 100mL of deionized water to obtain 10mg/mL of sodium caseinate stock solution;
5) dissolving 15mg curcumin powder in 100mL absolute ethanol to obtain 0.15mg/mL curcumin-ethanol solution, and storing at 4 deg.C in dark place for use;
6) at normal temperature, 15mL of 20mmol/L NaCl solution is placed on a magnetic stirrer with the rotating speed of 600r/min, 1mL of rice bran protein stock solution obtained in the step 2) and 1.5mL of sodium caseinate stock solution obtained in the step 4) are added while stirring, then adjusting the pH value of the mixed solution to 12.0 by using 1.0M sodium hydroxide, continuing to magnetically stir for 30min, adding 18mL of curcumin-ethanol solution obtained in the step 5) under dark condition, fully and uniformly mixing, adding 2mL of acacia gum stock solution obtained in step 3), stirring for 1h, removing ethanol with a rotary evaporator, slowly adjusting pH of the mixed system to 4.0 with 0.05M hydrochloric acid, centrifuging at 3000r/min for 10min, removing any insoluble large particles, taking centrifugal supernatant to obtain curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticle aqueous solution;
7) freezing the curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticle aqueous solution obtained in the step 6) at-20 ℃ for 24 hours, then placing the solution in a freeze dryer, and freeze-drying the solution at-0.5 mbar and-55 ℃ for 48 hours to obtain curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles.
The curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles prepared in the above examples 1 to 5 were detected, and the specific detection results were as follows:
as shown in fig. 1, the particle size distribution of the curcumin-loaded rice bran protein-gum arabic-sodium caseinate nanoparticles prepared in examples 1 to 5 is 363.6 to 521.6nm, and PDI is less than 0.3, which indicates that the nanoparticles prepared by the invention have small particle size, good dispersion degree and uniformity.
As shown in fig. 2, the Zeta potential of the curcumin-loaded rice bran protein-gum arabic-sodium caseinate nanoparticles prepared in examples 1 to 5 is between-17.4 mV and-19.6 mV, which indicates that the surfaces of the nanoparticles all have negative charges, and the curcumin-loaded rice bran protein-gum arabic-sodium caseinate nanoparticles prepared in example 3 have the most negative charges and the best stability.
As shown in fig. 3, the embedding rate of the curcumin-loaded rice bran protein-gum arabic-sodium caseinate nanoparticles prepared in examples 1 to 5 on curcumin is 67.1 to 91.9%, and the higher embedding rate indicates that the prepared nanoparticles can realize effective embedding on curcumin.
Fig. 4 is a scanning electron microscope image of the curcumin-loaded rice bran protein-gum arabic-sodium caseinate nanoparticles prepared in example 3, and the curcumin-loaded rice bran protein-gum arabic-sodium caseinate nanoparticles prepared in the invention are spherical and approximately spherical, and have smooth particle surfaces and uniform sizes.
Fig. 5 is a graph showing the effect of curcumin-loaded rice bran protein-gum arabic-sodium caseinate nanoparticles prepared in example 3 on the improvement of curcumin bioavailability. Bioavailability of curcumin in free curcumin and nanoparticles prepared in example 3 was evaluated by continuous in vitro digestion of simulated gastric fluid (60min) and intestinal fluid (120 min). The in vitro bioavailability of curcumin is considered to be the portion that is released by dissolution in gastrointestinal fluids and is generally absorbed only when it is released by dissolution in gastrointestinal fluids. Compared with free curcumin, the nanoparticles prepared in the example 3 have better solubility in the gastrointestinal tract and obviously improve the bioavailability of the curcumin.

Claims (9)

1. A preparation method of curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles is characterized by comprising the following steps:
1) mixing defatted rice bran with deionized water with the volume of 5 times, homogenizing the mixture for 3-5 min, stirring for 1-2 h at normal temperature, adjusting the pH of the mixture to 9.5 by using 3.0M sodium hydroxide, magnetically stirring for 1-2 h at 55 ℃, centrifuging for 30min at room temperature at 10000r/min, filtering the centrifuged supernatant with gauze, adjusting the pH of the filtrate to 4.4 by using 2.0M hydrochloric acid, standing for 30min, centrifuging for 30min at room temperature at 10000r/min, taking precipitate, washing the precipitate for 2 times by using deionized water to obtain a neutral protein solution, freezing for 24h at-20 ℃, then placing the neutral protein solution in a freeze dryer, and freeze-drying for 48h at-0.1-0.5 mbar, -60 ℃ to-50 ℃ to obtain rice bran protein;
2) dissolving the rice bran protein obtained in the step 1) in deionized water to prepare a 10mg/mL rice bran protein solution, and adjusting the pH of the solution by using 1.0M sodium hydroxide to obtain a rice bran protein stock solution;
3) at normal temperature, placing a NaCl solution on a magnetic stirrer with the rotation speed of 600r/min, adding the rice bran protein stock solution and the sodium caseinate stock solution obtained in the step 2) while stirring, then adjusting the pH of the mixed solution to 10.0-12.0 by using 1.0M sodium hydroxide, continuing to magnetically stir for 30min, adding a curcumin-ethanol solution under the dark condition, fully and uniformly mixing, adding an Arabic gum stock solution, continuing to stir for 1-2 h, rotationally evaporating to remove ethanol, slowly adjusting the pH of the mixed system by using 0.05M hydrochloric acid, finally centrifuging for 10min at 3000r/min, and taking a centrifuged supernatant to obtain a curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticle aqueous solution;
4) freezing the curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticle aqueous solution obtained in the step 3) at-20 ℃ for 24 hours, then placing the solution in a freeze dryer, and freeze-drying the solution at-60 ℃ to-50 ℃ for 48 hours at 0.1-0.5 mbar to obtain curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles.
2. The method according to claim 1, wherein the pH of the rice bran protein stock solution in the step 2) is 10.0 to 12.0.
3. The method according to claim 1, wherein in the step 3), the concentration of the NaCl solution is 5-25 mmol/L.
4. The method according to claim 1, wherein the sodium caseinate stock solution is prepared in step 3): sodium caseinate powder was dissolved in deionized water to give 10mg/mL sodium caseinate stock solution.
5. The preparation method according to claim 1, wherein in step 3), the curcumin-ethanol solution is prepared by: dissolving curcumin powder in anhydrous ethanol to obtain 0.15mg/mL curcumin-ethanol solution, and storing at 4 deg.C in dark place.
6. The method according to claim 1, wherein the gum arabic stock solution is prepared in step 3): acacia gum powder was dissolved in deionized water to give a 10mg/mL acacia gum stock solution.
7. The preparation method according to claim 1, wherein in the step 3), the NaCl solution, the rice bran protein stock solution, the sodium caseinate stock solution, the curcumin-ethanol solution and the Arabic gum stock solution are 15:1: 0.5-2.5: 18-21: 2 in volume ratio.
8. The preparation method according to claim 1, wherein in the step 3), the pH of the mixed system is 4.0 to 4.8.
9. The method for preparing curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles as claimed in any one of claims 1 to 8, wherein the particle size of the curcumin-loaded rice bran protein-Arabic gum-sodium caseinate nanoparticles is 310.2-1000 nm, the PDI is 0.136-0.491, the Zeta potential is-14.2-20.2 mV, the curcumin embedding rate is 58.3-98.5%, the bioavailability of curcumin in simulated gastric juice is 15.1-18.3%, and the bioavailability in simulated intestinal juice is 23.1-27.2%.
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