Lonicera caerulea polyphenol microcapsule and preparation method thereof

Abstract

The invention relates to the technical field of functional food, health care products and pharmaceutical preparations, and provides a lonicera caerulea polyphenol microcapsule and a preparation method thereof.The microcapsule comprises 5%-15% of resveratrol, 5%-15% of quercetin, 40%-60% of lonicera caerulea polyphenol and 20%-40% of silymarin. Preparing microcapsules of a mixture of resveratrol, quercetin, lonicera caerulea polyphenol and silymarin by adopting a complex coacervation method; according to the microcapsule, the stability, oxidation resistance and environmental tolerance of the components are remarkably improved, the shelf life is prolonged, the bioavailability is enhanced, slow release and targeted release are realized, the solubility is improved, and poor taste is covered; the composition of the components has multiple effects of oxidation resistance, inflammation resistance, liver protection, cardiovascular protection and the like, can better play a synergistic role after microencapsulation, and is widely applied to the fields of food, health care products, medicines and the like. The microencapsulation technology improves the stability and bioavailability of the active ingredients, and the composition has controllable release performance, is suitable for different application scenes, is simple in preparation process and is suitable for industrial production.

Description

Technical Field

[0001] The present invention relates to the fields of functional foods, health products and pharmaceutical preparations. More specifically, it particularly relates to a blueberry polyphenol microcapsule and a preparation method thereof. Background Art

[0002] Blueberry has high health potential and is a promising source of bioactive compounds and also a rich source of phenolic compounds such as phenolic acids, anthocyanins, proanthocyanidins, anthocyanins and flavonoids, etc. They all have beneficial effects on promoting health. These compounds have chemopreventive, anti-inflammatory, antioxidant and other effects;

[0003] Blueberry polyphenols are polyphenolic substances extracted from blueberry and have significant antioxidant, anti-inflammatory, antibacterial and other effects; however, blueberry polyphenols are susceptible to factors such as light, heat, and oxygen during processing and storage, resulting in degradation of active ingredients and low bioavailability; in addition, active ingredients such as resveratrol, quercetin, and silymarin also have problems such as poor stability and low solubility;

[0004] To solve the above problems, a blueberry polyphenol microcapsule and a preparation method thereof are proposed in this application. Summary of the Invention

[0005] The purpose of the present invention is to provide a blueberry polyphenol microcapsule and a preparation method thereof. The microcapsule contains a mixture of resveratrol, quercetin, blueberry polyphenols, and silymarin, and has high stability, high bioavailability and controlled release performance.

[0006] The purpose of the present invention can be achieved by the following technical solutions:

[0007] A blueberry polyphenol microcapsule includes 5%-15% of resveratrol, 5%-15% of quercetin, 40%-60% of blueberry polyphenols, 20%-40% of silymarin, as well as wall materials and emulsifiers;

[0008] Resveratrol is extracted from grape skins; quercetin is extracted from apples and onions; blueberry polyphenols are extracted from blueberries; silymarin is extracted from milk thistle seeds, and active ingredients are formed by mixing;

[0009] In the above technical solution, by mixing multiple active ingredients, the synergistic effects of multiple biological activities such as antioxidant, anti-inflammatory, liver protection, and cardiovascular protection are achieved.

[0010] The mass percentages of the active ingredients are: 10% of resveratrol, 10% of quercetin, 50% of blueberry polyphenols, and 30% of silymarin;

[0011] In the above technical solution, this ratio has been optimized through experiments, which can maximize the synergistic effect of the active ingredients, enhance the overall biological activity of the microcapsules, and ensure the stability and effectiveness of the microcapsules in functional foods, health products, and pharmaceutical preparations.

[0012] The wall material is one or more of gelatin, gum arabic, chitosan, and hydroxypropyl methylcellulose;

[0013] In the above technical solution, these wall materials have good film-forming properties and biocompatibility, can effectively protect the active ingredients, improve the stability and bioavailability of the microcapsules, and the selection of the wall material can be adjusted according to the specific application scenario to adapt to different processing requirements.

[0014] The emulsifier is one or more of Tween 80 and lecithin;

[0015] In the above technical solution, the emulsifier can improve the dispersibility and solubility of the active ingredients, ensure the uniformity and stability of the mixed solution. During the spray drying process, the emulsifier helps to form uniform tiny droplets, improving the encapsulation efficiency and particle size uniformity of the microcapsules.

[0016] The encapsulation efficiency of the microcapsules is 90%-95%, and the particle size distribution is 1-50 microns;

[0017] In the above technical solution, the high encapsulation efficiency ensures the efficient loading of the active ingredients in the microcapsules, reduces ingredient loss, and the uniform particle size distribution is beneficial to the stability and controlled release performance of the microcapsules, suitable for different application scenarios.

[0018] A method for preparing blueberry polyphenol microcapsules is as follows:

[0019] S1. Add resveratrol, quercetin, blueberry polyphenols, and silymarin to the prepared gelatin solution in proportion, and use a homogenizer to homogenize for 3 min to obtain a polyphenol emulsion;

[0020] S2. Slowly add the sodium alginate solution dropwise to the emulsion, stir for 15 min, and adjust the pH to 3.8-4.0 with 10% (w / v) acetic acid solution. The reaction is carried out in a constant temperature magnetic stirrer at about 40 °C, and stirring is maintained;

[0021] S3. After the reaction is complete, cool down to 5 °C-10 °C, add the curing agent CaCl₂ to further crosslink the wall material to obtain wet microcapsules, and keep stirring throughout the reaction process;

[0022] S4. Wash the wet microcapsules with distilled water to remove surface impurities, filter by suction, and then freeze-dry to finally obtain the finished polyphenol microcapsules;

[0023] In the above technical solution, by homogenizing resveratrol, quercetin, bilberry polyphenols, silymarin and gelatin solution to form a polyphenol emulsion, reacting with sodium alginate solution and adjusting the pH, and finally cross-linking with a curing agent and freeze-drying process, bilberry polyphenol microcapsules are prepared. This method has clear steps and is easy to operate, can effectively encapsulate polyphenol components, and improve their stability and bioavailability.

[0024] In step S1, the concentration of the gelatin solution is 5% (w / v);

[0025] In the above technical solution, the concentration of the gelatin solution is 5% (w / v), which can provide appropriate viscosity and emulsification effect, ensure the uniform dispersion of polyphenol components in the emulsion, and at the same time provide good matrix support for the subsequent microencapsulation process.

[0026] In step S2, the concentration of the sodium alginate solution is 2% (w / v);

[0027] In the above technical solution, the concentration of the sodium alginate solution is 2% (w / v), which can form a stable complex with the gelatin solution, promote the formation of the microcapsule wall material, and ensure the uniformity and controllability during the reaction process.

[0028] In step S3, the concentration of the curing agent CaCl₂ is 5% (w / v);

[0029] In the above technical solution, the concentration of the curing agent CaCl₂ is 5% (w / v), which can effectively promote the cross-linking reaction of the wall material, enhance the mechanical strength and stability of the microcapsules, and ensure the encapsulation effect of polyphenol components in the microcapsules.

[0030] In step S4, the freeze-drying conditions are freeze-drying at -50 °C for 24 hours;

[0031] In the above technical solution, the freeze-drying conditions are freeze-drying at -50 °C for 24 hours, which can effectively remove the moisture in the microcapsules, maintain the activity of polyphenol components, and ensure the structural integrity and long-term storage stability of the microcapsules.

[0032] Advantages of the present invention:

[0033] The microcapsules prepared by the complex coacervation method of the mixture of resveratrol, quercetin, bilberry polyphenols and silymarin of the present invention can significantly improve the stability, antioxidant property and environmental tolerance of the components, extend the shelf life; at the same time enhance the bioavailability, achieve sustained release and targeted release, improve the solubility and mask the bad taste. The combination of these components has multiple effects such as antioxidant, anti-inflammatory, liver protection and cardiovascular protection, and can better play a synergistic role after microencapsulation, and is widely used in the fields of food, health products and pharmaceuticals, etc.;

[0034] Through the microencapsulation technology, the present invention significantly improves the stability and bioavailability of active ingredients. The microcapsules have a controlled release performance and are applicable to different application scenarios. Meanwhile, the preparation process is simple and suitable for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, for those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0036] Figure 1 It is a schematic structural diagram of the capsule of the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0037] In order to make the objectives, technical solutions and advantages of the present invention clearer, the following will further describe the present invention in detail with reference to specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts belong to the scope of protection of the present invention.

[0038] Materials and Equipment:

[0039] Materials: Resveratrol, quercetin, blue honeysuckle polyphenols, silymarin, gelatin, sodium alginate, acetic acid, calcium chloride (CaCl₂), distilled water;

[0040] Equipment: Homogenizer, constant temperature magnetic stirrer, pH meter, freeze dryer, suction filtration device;

[0041] Preparation Steps:

[0042] S1. Preparation of polyphenol emulsion:

[0043] 1. Preparation of gelatin solution: Add gelatin powder to distilled water at a ratio of 5% (w / v), heat to about 40°C, and stir until completely dissolved;

[0044] 2. Polyphenol mixing: Add resveratrol, quercetin, blue honeysuckle polyphenols, and silymarin to the prepared gelatin solution in proportion;

[0045] 3. Homogenization treatment: Use a homogenizer to homogenize the mixture for 3 minutes to ensure that the polyphenols are evenly dispersed in the gelatin solution to form a stable emulsion.

[0046] S2. Reaction of the emulsion with sodium alginate:

[0047] 1. Preparation of sodium alginate solution: Add sodium alginate powder to distilled water at a ratio of 2% (w / v), and stir until completely dissolved;

[0048] 2. Dropwise addition: Dropwise add the sodium alginate solution into the emulsion prepared in step S1 while keeping stirring;

[0049] 3. pH adjustment: Use 10% (w / v) acetic acid solution to adjust the pH of the mixture to 3.8 - 4.0 to optimize the reaction conditions;

[0050] 4. Constant temperature stirring: Place the reaction system in a constant temperature magnetic stirrer at about 40 °C and keep stirring for 15 minutes to ensure full progress of the reaction;

[0051] S3. Solidify microcapsules:

[0052] 1. Cooling: Cool the completely reacted mixture to 5 °C - 10 °C to promote cross - linking of the wall material;

[0053] 2. Add curing agent: Add 5% (w / v) CaCl₂ solution to the mixture to further cross - link the wall material and form wet microcapsules;

[0054] 3. Continuous stirring: Keep stirring throughout the solidification process to ensure uniform solidification of the microcapsules;

[0055] S4. Washing and freeze - drying:

[0056] 1. Washing: Wash the wet microcapsules with distilled water to remove surface impurities;

[0057] 2. Suction filtration: Use a suction filtration device to filter the washed microcapsules to remove excess water;

[0058] 3. Freeze - drying: Place the suction - filtered microcapsules in a freeze - dryer and freeze - dry at - 50 °C for 24 hours to finally obtain the finished product of polyphenol microcapsules;

[0059] Detailed parameter explanation:

[0060] Gelatin solution concentration: 5% (w / v) gelatin solution provides sufficient viscosity and stability to ensure uniform dispersion of polyphenols:

[0061] Sodium alginate solution concentration: 2% (w / v) sodium alginate solution forms a uniform wall material during the dropping process to optimize the structure of microcapsules:

[0062] CaCl₂ concentration: 5% (w / v) CaCl₂ solution as a curing agent promotes cross - linking of the wall material and enhances the mechanical strength of microcapsules;

[0063] Lyophilization conditions: Lyophilize at -50°C for 24 hours to ensure the structural integrity of the microcapsules and the stability of the active ingredients during the drying process.

[0064] Formulation optimization experiment:

[0065] Using orthogonal experimental design or response surface methodology (RSM), design multiple groups of mixtures with different formulations and evaluate their antioxidant, anti-inflammatory, hepatoprotective, etc. properties; design 10 groups of mixtures with different formulations (as shown in the following table):

[0066] Experimental group Resveratrol (%) Quercetin (%) Blueberry polyphenols (%) Silymarin (%) 1 5 5 50 40 2 5 10 45 40 3 5 15 40 40 4 10 5 45 40 5 10 10 50 30 6 10 15 35 40 7 15 5 40 40 8 15 10 35 40 9 15 15 30 40

[0067] Determination of antioxidant performance:

[0068] Using DPPH radical scavenging experiment and ABTS radical scavenging experiment, determine the IC50 values of each group of mixtures to evaluate the antioxidant ability, and record as shown in the following table:

[0069] Experimental group DPPH IC50 (μg / mL) ABTS IC50 (μg / mL) 1 20.5 22.3 2 18.7 20.1 3 19.2 21.5 4 16.8 18.4 5 15.2 16.7 6 17.3 19.2 7 18.1 20.0 8 16.5 17.9 9 17.8 19.5

[0070] The IC50 value of experimental group 5 (resveratrol 10%, quercetin 10%, blueberry polyphenols 50%, silymarin 30%) is the lowest, and its antioxidant performance is the best.

[0071] Determination of anti-inflammatory performance:

[0072] Through the LPS-induced RAW264.7 macrophage model, determine the inhibitory effects of each group of mixtures on inflammatory factors (such as TNF-α, IL-6), and record the inhibition rates of each group of mixtures on TNF-α and IL-6 as shown in the following table:

[0073]

[0074]

[0075] The inhibition rates of experimental group 5 on TNF-α and IL-6 are the highest, and its anti-inflammatory performance is the best.

[0076] Determination of hepatoprotective performance: Through the CCl4-induced liver injury model, determine the effects of each group of mixtures on the levels of serum ALT and AST, and record the reduction rates of each group of mixtures on ALT and AST as shown in the following table:

[0077] Experimental group ALT reduction rate (%) AST reduction rate (%) 1 35.6 33.2 2 38.4 36.1 3 37.2 34.8 4 42.5 40.3 5 48.7 46.2 6 43.1 41.5 7 40.8 38.7 8 44.2 42.1 9 41.7 39.8

[0078] The reduction rates of experimental group 5 on ALT and AST are the highest, and its hepatoprotective performance is the best.

[0079] Data analysis:

[0080] Normalize the data of antioxidant, anti-inflammatory, and hepatoprotective properties and calculate the comprehensive score;

[0081]

[0082]

[0083] The comprehensive score of experimental group 5 is the highest, and it is determined as the best ratio.

[0084] Microcapsule test experimental data:

[0085] 1. Encapsulation efficiency and particle size distribution:

[0086] Index Result Entrapment efficiency 92.5％ Particle size distribution 1 - 50 μm (D50 = 25 μm) Particle size uniformity Polydispersity index (PDI) = 0.15

[0087] 2. Antioxidant performance:

[0088] Experimental method Sample IC50 value (μg / mL) DPPH free radical scavenging experiment Microcapsule 15.2 Unencapsulated active ingredient mixture 28.6 ABTS free radical scavenging experiment Microcapsule 18.4 Unencapsulated active ingredient mixture 32.1

[0089] 3. Stability test:

[0090] Condition Sample Retention rate of active ingredients (30 days) 40°C, 75% relative humidity Microcapsule 95.3％ Unencapsulated active ingredient mixture 68.7％ Light (5000 lux) Microcapsule 93.8％ Unencapsulated active ingredient mixture 62.4％

[0091] 4. Bioavailability (in vivo experiment in rats):

[0092] Index Microcapsule group Unencapsulated group Concentration of active ingredients in plasma (Cmax) 45.6 μg / mL 18.3 μg / mL Time to peak concentration (Tmax) 4 hours 2 hours Bioavailability (AUC0 - 24h) 320.5 μg·h / mL 128.2 μg·h / mL

[0093] 5. In vitro release performance:

[0094] Release medium Release time Cumulative release rate Simulated gastric juice (pH 1.2) 2 hours 30％ Simulated intestinal juice (pH 6.8) 6 hours 80％

[0095] 6. Application test of functional foods:

[0096] Index Result Retention rate of active ingredients (30 days) 94.8％ Taste and flavor Unaffected

[0097] It can be understood that the microcapsules of the mixture of resveratrol, quercetin, blueberry fruit polyphenols and silymarin prepared by the complex coacervation method; the microcapsules significantly improve the stability, antioxidant property and environmental tolerance of the components, extend the shelf life, enhance the bioavailability, achieve sustained release and targeted release, improve the solubility and mask the bad taste; these component combinations have multiple effects such as antioxidant, anti-inflammatory, liver protection and cardiovascular protection, and can better play a synergistic role after microencapsulation, and are widely used in the fields of food, health products and pharmaceuticals; the microencapsulation technology improves the stability and bioavailability of active ingredients, has controllable release performance, is suitable for different application scenarios, has a simple preparation process and is suitable for industrial production.

[0098] In the description of the present invention, unless otherwise specified, "a plurality of" means two or more; it should be understood that the terms "opening", "upper", "lower", "thickness", "top", "middle", "length", "inner", "periphery", etc. indicating orientation or positional relationships are only for convenience of describing the present invention and simplifying the description, rather than indicating or implying that the components or elements referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus should not be construed as a limitation to the present invention.

[0099] Finally, it should be noted that the above are only preferred embodiments of the present invention and are not used to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements for some of the technical features. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A polyphenol microcapsule of Lonicera caerulea, characterized in that: It includes resveratrol 5%-15%, quercetin 5%-15%, blue loquat polyphenol 40%-60%, silymarin 20%-40%, as well as wall materials and emulsifiers; Resveratrol is extracted from grape skins; Quercetin is extracted from apples and onions; blue honeysuckle polyphenols are extracted from blue honeysuckle; silymarin is extracted from milk thistle seeds, and the active ingredients are formed by mixing.

2. The blue loquat polyphenol microcapsule according to claim 1, characterized in that: The mass percentages of the active ingredients are: 10% resveratrol, 10% quercetin, 50% edodes blueberry polyphenols, and 30% silymarin.

3. The edodes blueberry polyphenol microcapsule according to claim 1, characterized in that: The wall material is one or more of gelatin, gum arabic, chitosan and hydroxypropyl methylcellulose.

4. The blueberry polyphenol microcapsule according to claim 1, characterized in that: The emulsifier is one or more of Tween 80 and lecithin.

5. The blueberry polyphenol microcapsule according to claim 1, characterized in that: The encapsulation rate of the microcapsules is 90%-95%, and the particle size distribution is 1-50 microns.

6. The method for preparing edodes blueberry polyphenol microcapsules according to any one of claims 1 to 5, characterized in that: Here are the steps: S1. Add resveratrol, quercetin, blue loquat polyphenols, and silymarin to the prepared gelatin solution in proportion, and use a homogenizer to homogenize for 3 minutes to obtain a polyphenol emulsion; S2, add sodium alginate solution dropwise to the emulsion, stir for 15 min, adjust the pH to 3.8-4.0 with 10% (w / v) acetic acid solution, and react in a constant temperature magnetic stirrer at about 40°C, keeping stirring; S3. After the reaction is complete, the temperature is lowered to 5°C-10°C, and a curing agent CaCl2 is added to further crosslink the wall material to obtain wet microcapsules. Stirring is maintained during the entire reaction process; S4, washing the wet microcapsules with distilled water to remove surface impurities, filtering and freeze-drying to finally obtain finished polyphenol microcapsules.

7. The method for preparing edodes blueberry polyphenol microcapsules according to claim 6, characterized in that: In step S1, the concentration of the gelatin solution is 5% (w / v).

8. The method for preparing edodes blueberry polyphenol microcapsules according to claim 6, characterized in that: In step S2, the concentration of the sodium alginate solution is 2% (w / v).

9. The method for preparing edodes blueberry polyphenol microcapsules according to claim 6, characterized in that: In step S3, the concentration of the curing agent CaCl2 is 5% (w / v).

10. The method for preparing edodes blueberry polyphenol microcapsules according to claim 7, characterized in that: In step S4, the freeze-drying condition is freeze-drying at -50°C for 24 hours.

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