Preparation method of soybean 11S protein-high-ester citrus pectin liposome hydrogel
Soybean 11S protein-high ester citrus pectin liposome hydrogel was prepared by film hydration, which solved the problems of liposome stability and bioavailability, achieved the improvement of liposome stability and bioavailability, and extended the action time of active substances.
Patent Information
- Application Number
- CN202411751855.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-02
- Publication Date
- 2025-03-28
AI Technical Summary
Existing liposomes are prone to fusion and aggregation during storage and application, and encapsulated functional substances are prone to leakage. They are not suitable for direct oral consumption, making it difficult to solve the problems of their stability and bioavailability.
Soybean 11S protein-high ester citrus pectin liposome hydrogel was prepared by thin-film hydration method, and the hydrogel network was formed through Ca2+ induction, enhancing the stability and biocompatibility of liposomes.
The stability and bioavailability of liposomes are improved, the time of action of active substances in the receptor is extended, and the stability of the drug and the concentration of the site of action are improved.
Smart Images
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Figure HDA0005165194090000012
Abstract
Description
Technical Field
[0001] The invention belongs to the field of liposome development, and specifically relates to a method for preparing soybean 11S protein-high-ester citrus pectin liposome hydrogel. Background Art
[0002] Liposomes are closed vesicles composed of phospholipid bilayers, and the structure is very similar to biological membranes, with the ability to embed water-soluble or fat-soluble substances, so liposomes have good biocompatibility and biodegradability. Using liposomes to encapsulate substances can reduce their drug toxicity to non-target organs, and can also improve the stability and bioavailability of the encapsulated substances. However, the properties of liposomes themselves are not stable, they are easily oxidized, and there are problems such as poor stability in external gastrointestinal digestion. Therefore, liposomes are not suitable for direct oral consumption, and fusion aggregation is prone to occur during storage and application, and the encapsulated functionalized substances are also prone to leakage. These problems cannot be solved by improving the preparation technology alone. Therefore, it is necessary to further improve the performance of liposomes through surface modification methods.
[0003] The surface modification of liposomes is to use physical or chemical means to adsorb bioactive substances on the surface of liposomes through electrostatic effects, hydrophobic interactions and other forces, forming a shell-core structure to prevent the liposomes from decomposing and breaking in the environment. With the diversification of dietary habits, nutrients carried by liposomes face more challenges. Liposome hydrogels have attracted widespread attention because their three-dimensional spatial network can effectively protect liposomes, reduce damage in the human body and leakage of bioactive substances. Compared with the use of hydrogels or liposomes alone, loading liposomes into hydrogels can make bioactive compounds have high stability and controlled release behavior, and can form a stable three-dimensional spatial structure. Liposome hydrogel drug delivery systems have been proven to be able to double-layer controlled release of drugs, improve drug stability and drug concentration at the site of action, and are conducive to improving drug bioavailability.
[0004] Zhong Wei et al. prepared resveratrol liposome gel to treat melasma, improve antioxidant capacity, and reduce local damage. However, there are still few reports on liposome hydrogels in the food field. Summary of the invention
[0005] The purpose of the present invention is to provide a stable and non-toxic soybean 11S protein-high-ester citrus pectin liposome hydrogel system for delivering active substances and prolonging their action time in receptors.
[0006] In order to achieve the above object, the present invention adopts the following technical solutions:
[0007] The preparation method of the high-ester citrus pectin liposome hydrogel system adopts a thin film hydration method, which specifically includes the following steps:
[0008] (1) Soybean lecithin (1 g) and cholesterol (0.1 g) were dissolved in (50 mL) of ethanol solution and rotary evaporated at 40°C until a thin film was formed. The liposome membrane on the wall of the bottle was dissolved with (100 mL) of distilled water to obtain a crude liposome solution (10 mg / mL);
[0009] (2) using a cell disruptor to sonicate the liposome solution at 300 W for 10 min to obtain a liposome solution of uniform size (10 mg / mL);
[0010] (3) adding the liposome solution to the high-ester citrus pectin solution to obtain high-ester citrus pectin liposomes;
[0011] (4) Dissolve soybean 11S protein in water (100 mL) at a concentration of 7%;
[0012] (5) Weigh the high-ester citrus pectin liposome aqueous solution prepared in step (3), add it to the gel prepared in step (4), and use Ca 2+ The hydrogel network is induced to form, thus obtaining soybean 11S protein-high-ester citrus pectin liposome hydrogel.
[0013] It is further defined that in step (1), 1 g of soybean lecithin and 0.1 g of cholesterol;
[0014] It is further defined that in step (3), the high-ester citrus pectin solution is 100 mL, the concentration is 0.5%, and the ratio of the high-ester citrus pectin solution to the liposome solution is 1:1;
[0015] It is further defined that the soybean 11S protein concentration in step (4) is 7%;
[0016] Beneficial effects: The present invention firstly prepares high-ester citrus pectin liposomes by thin film hydration method. Compared with naked liposomes, high-ester citrus pectin has a high degree of methoxylation, and the formed hydrogel has stronger stability, and can bind and stabilize functional ingredients, such as drugs, nutrients, etc., to improve their delivery efficiency and bioavailability in the body. BRIEF DESCRIPTION OF THE DRAWINGS
[0017] Figure 1 Schematic diagram of the average particle size of high-ester citrus pectin-liposomes with different concentrations;
[0018] Figure 2 Soybean 11S protein-high ester citrus pectin liposome gel (Lip-CP-11S) with different concentrations of high ester citrus pectin DETAILED DESCRIPTION
[0019] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the present invention are clearly and completely described below in combination with the embodiments of the present invention. The described embodiments are part of the embodiments of the present invention, not all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work belong to the protection scope of the present invention.
[0020] Lecithin and cholesterol were purchased from Shanghai Yuanye Biotechnology Co., Ltd.
[0021] High ester citrus pectin was purchased from Shandong Andeli Group Co., Ltd.
[0022] The soybeans selected in the present invention are Longken soybeans, which are purchased commercially.
[0023] Embodiment 1:
[0024] A method for preparing soybean 11S protein-high-ester citrus pectin liposome hydrogel, the specific steps are as follows:
[0025] (1) Dissolve 1 g of soybean lecithin and 0.1 g of cholesterol in 50 mL of ethanol solution, evaporate the ethanol using a rotary evaporator at 40°C until the ethanol is evaporated to dryness to form a thin film, add 100 mL of distilled water to the rotary evaporator to dissolve the liposome film to obtain crude liposomes of 10 mg / mL. Use a cell disruptor to sonicate at 300 W for 10 min to obtain a liposome solution of uniform size.
[0026] (2) Weigh 0.1 g of high-ester citrus pectin and add it to 100 mL of distilled water (pH 3.5) to obtain a 1 mg / mL high-ester citrus pectin solution. Add an equal volume of the liposome solution in (1) to the high-ester citrus pectin solution to obtain a high-ester citrus pectin-liposome solution. Use a freeze dryer to freeze-dry to obtain a high-ester citrus pectin-liposome powder sample.
[0027] (3) Weigh 7 g of soybean 11S protein and add it to 100 mL of distilled water to obtain a 70 mg / mL soybean 11S protein solution. Add Ca 2+ The mixture was heated at 85°C for 30 min, during which high-ester citrus pectin-liposome powder was added to finally form soybean 11S protein-high-ester citrus pectin liposome hydrogel.
[0028] Embodiment 2:
[0029] A method for preparing soybean 11S protein-high-ester citrus pectin liposome hydrogel, the specific steps are as follows:
[0030] (1) Dissolve 1 g of soybean lecithin and 0.1 g of cholesterol in 50 mL of ethanol solution, evaporate the ethanol using a rotary evaporator at 40°C until the ethanol is evaporated to dryness to form a thin film, add 100 mL of distilled water to the rotary evaporator to dissolve the liposome film to obtain crude liposomes of 10 mg / mL. Use a cell disruptor to sonicate at 300 W for 10 min to obtain a liposome solution of uniform size.
[0031] (2) Weigh 0.2 g of high-ester citrus pectin and add it to 100 mL of distilled water (pH 3.5) to obtain a 2 mg / mL high-ester citrus pectin solution. Add an equal volume of the liposome solution in (1) to the high-ester citrus pectin solution to obtain a high-ester citrus pectin-liposome solution. Use a freeze dryer to freeze-dry to obtain a high-ester citrus pectin-liposome powder sample.
[0032] (3) Weigh 7 g of soybean 11S protein and add it to 100 mL of distilled water to obtain a 70 mg / mL soybean 11S protein solution. Add Ca 2+ The mixture was heated at 85°C for 30 min, during which high-ester citrus pectin-liposome powder was added to finally form soybean 11S protein-high-ester citrus pectin liposome gel.
[0033] Embodiment 3:
[0034] A method for preparing soybean 11S protein-high-ester citrus pectin liposome hydrogel, the specific steps are as follows:
[0035] (1) Dissolve 1 g of soybean lecithin and 0.1 g of cholesterol in 50 mL of ethanol solution, evaporate the ethanol using a rotary evaporator at 40°C until the ethanol is evaporated to dryness to form a thin film, add 100 mL of distilled water to the rotary evaporator to dissolve the liposome film to obtain crude liposomes of 10 mg / mL. Use a cell disruptor to sonicate at 300 W for 10 min to obtain a liposome solution of uniform size.
[0036] (2) Weigh 0.3 g of high-ester citrus pectin and add it to 100 mL of distilled water (pH 3.5) to obtain a 3 mg / mL high-ester citrus pectin solution. Add an equal volume of the liposome solution in (1) to the high-ester citrus pectin solution to obtain a high-ester citrus pectin-liposome solution. Use a freeze dryer to freeze-dry to obtain a high-ester citrus pectin-liposome powder sample.
[0037] (3) Weigh 7 g of soybean 11S protein and add it to 100 mL of distilled water to obtain a 70 mg / mL soybean 11S protein solution. Add Ca 2+ The mixture was heated at 85°C for 30 min, during which high-ester citrus pectin-liposome powder was added to finally form soybean 11S protein-high-ester citrus pectin liposome hydrogel.
[0038] Embodiment 4:
[0039] A method for preparing soybean 11S protein-high-ester citrus pectin liposome hydrogel, the specific steps are as follows:
[0040] (1) Dissolve 1 g of soybean lecithin and 0.1 g of cholesterol in 50 mL of ethanol solution, evaporate the ethanol using a rotary evaporator at 40°C until the ethanol is evaporated to dryness to form a thin film, add 100 mL of distilled water to the rotary evaporator to dissolve the liposome film to obtain crude liposomes of 10 mg / mL. Use a cell disruptor to sonicate at 300 W for 10 min to obtain a liposome solution of uniform size.
[0041] (2) Weigh 0.4 g of high-ester citrus pectin and add it to 100 mL of distilled water (pH 3.5) to obtain a 4 mg / mL high-ester citrus pectin solution. Add an equal volume of the liposome solution in (1) to the high-ester citrus pectin solution to obtain a high-ester citrus pectin-liposome solution. Use a freeze dryer to freeze-dry to obtain a high-ester citrus pectin-liposome powder sample.
[0042] (3) Weigh 7 g of soybean 11S protein and add it to 100 mL of distilled water to obtain a 70 mg / mL soybean 11S protein solution. Add Ca 2+ The mixture was heated at 85°C for 30 min, during which high-ester citrus pectin-liposome powder was added to finally form soybean 11S protein-high-ester citrus pectin liposome hydrogel.
[0043] Embodiment 5:
[0044] A method for preparing soybean 11S protein-high-ester citrus pectin liposome hydrogel, the specific steps are as follows:
[0045] (1) Dissolve 1 g of soybean lecithin and 0.1 g of cholesterol in 50 mL of ethanol solution, evaporate the ethanol using a rotary evaporator at 40°C until the ethanol is evaporated to dryness to form a thin film, add 100 mL of distilled water to the rotary evaporator to dissolve the liposome film to obtain crude liposomes of 10 mg / mL. Use a cell disruptor to sonicate at 300 W for 10 min to obtain a liposome solution of uniform size.
[0046] (2) Weigh 0.5 g of high-ester citrus pectin and add it to 100 mL of distilled water (pH 3.5) to obtain a 5 mg / mL high-ester citrus pectin solution. Add an equal volume of the liposome solution in (1) to the high-ester citrus pectin solution to obtain a high-ester citrus pectin-liposome solution. Use a freeze dryer to freeze-dry to obtain a high-ester citrus pectin-liposome powder sample.
[0047] (3) Weigh 7 g of soybean 11S protein and add it to 100 mL of distilled water to obtain a 70 mg / mL soybean 11S protein solution. Add Ca 2+The mixture was heated at 85°C for 30 min, during which high-ester citrus pectin-liposome powder was added to finally form soybean 11S protein-high-ester citrus pectin liposome hydrogel.
Claims
1. A soybean 11S protein-high-ester citrus pectin liposome hydrogel, characterized in that: After the high-ester citrus pectin liposome system is formed, it is added into soybean 11S protein hydrogel to obtain a gel system.
2. The method for preparing the high-ester citrus pectin liposome hydrogel system according to claim 1, characterized in that: The specific steps include: Step 1, liposome preparation Step 1.1, dissolve 1 g of soybean lecithin and 0.1 g of cholesterol in 50 mL of ethanol solution, rotary evaporate the ethanol solution at 40°C, and dissolve the liposome membrane with 100 mL of distilled water to obtain a 10 mg / mL crude liposome solution; Step 1.2, using a cell disruptor to sonicate the liposome solution at 300 W for 10 min to obtain liposomes of uniform size; Step 2, Preparation of high-ester citrus pectin liposomes Step 2.1, adding the liposome solution to the same volume of high-ester citrus pectin (CP) solution (10 mg / mL) to obtain Lip-CP solution; Step 2.2, adding according to the ratio concentration of 1:(1, 2, 3, 4, 5) to obtain Lip-CP 0.1%, Lip-CP 0.2%, Lip-CP 0.3%, Lip-CP 0.4%, Lip-CP 0.5%; Step 3, preparation of soybean 11S protein-high ester citrus pectin liposome hydrogel Step 3.1, dissolving soybean 11S protein in water at a concentration of 7%; Step 3.2, add the optimal ratio of high-ester citrus pectin liposomes from the above part and use Ca 2+ The hydrogel network was induced to form and soybean 11S protein-liposome hydrogel was prepared.
3. The preparation method according to claim 2, characterized in that: The high ester citrus pectin (0.4 g) described in step 2.
1.
4. The preparation method according to claim 2, characterized in that: Soybean 11S protein (7 g) as described in step 3.
1.
5. The method for preparing high-ester citrus pectin liposome hydrogel according to claim 2 is used to prepare soybean 11S protein-high-ester citrus pectin liposome hydrogel.