EPA liposome as well as preparation method and application thereof

Abstract

The invention discloses an EPA liposome as well as a preparation method and application thereof, and particularly relates to the technical field of cosmetics. The EPA liposome disclosed by the invention is prepared by carrying out liposome self-assembly on the following raw materials in parts by mass: 1 to 4 parts of microalgae oil EPA, 1 to 2 parts of epicatechin-3-gallate, 0.2 to 1 part of alpha-tocopherol, 0.02 to 0.1 part of ginsenoside Rg3, 2 to 10 parts of lipid, 1 to 10 parts of an emulsifier and the balance of water, wherein the total amount is 100 parts. According to the EPA liposome disclosed by the invention, the stable effect of EPA in composite systems such as cosmetics and skin care products is achieved through the cooperation of functional substances contained in the EPA liposome and the entrapment design of the nano liposome, meanwhile, the in-situ anti-oxidation and protection functions of EPA are enhanced, the auto-oxidation of EPA is slowed down, and the improvement of bioavailability and the enhancement of aging delaying effect are realized.

Description

Technical Field

[0001] The present invention belongs to the technical field of cosmetics, and particularly relates to an EPA liposome, a preparation method thereof and an application thereof. Background Art

[0002] Algal oil EPA belongs to ω-3 unsaturated fatty acids, and can reduce oxidative damage, improve endothelial function, alleviate inflammation and provide in-situ antioxidant and protection functions by enhancing mitochondrial function and biosynthesis, and is expected to be used in cosmetics to relieve skin aging. However, EPA is extremely susceptible to the influence of light, oxygen, overheating, metal elements and free radicals, causing physical and chemical instabilities such as oxidation, rancidity and polymerization, which prevent its direct application in the compound system of cosmetics and skin care products. Therefore, designing and preparing an EPA preparation with stable efficacy in the compound system of cosmetics and skin care products has become a hot topic.

[0003] Currently, to improve the instability problem of EPA, the related technologies disclose a technical strategy for embedding and disposing EPA through encapsulation technology. For example, the patent application with publication number CN118542456A designs and prepares an emulsion for encapsulating EPA with polysaccharide and citrus fiber as composite wall materials, which enhances the protection and delivery effects on EPA, extends the shelf life, and improves the physical and chemical stability of the EPA carrier.

[0004] However, common encapsulation technologies such as the above still have deficiencies in practical applications. For example, the auxiliary acidity regulators, antioxidants and other compounding substances are generally limited to improving the stability of the encapsulation wall layer, resulting in poor biological efficacy of the EPA carrier, which greatly affects the achievement of the in-situ antioxidant and protection functions of EPA. Summary of the Invention

[0005] The present invention provides an EPA liposome, a preparation method thereof and an application thereof, aiming to solve the problem of poor biological efficacy of the existing EPA carrier.

[0006] To achieve the above object, the present application discloses the following technical solutions:

[0007] The first aspect of the present invention provides an EPA liposome, which is prepared and formed by self-assembly of liposomes through the following raw materials in parts by mass:

[0008] 1-4 parts of microalgal oil EPA, 1-2 parts of epicatechin-3-gallate, 0.2-1 part of α-tocopherol, ginsenoside Rg 3 0.02-0.1 part, 2-10 parts of lipid, 1-10 parts of emulsifier, and water is made up to 100 parts.

[0009] In a possible implementation, the EPA liposome is prepared and formed by self-assembly of liposomes through the following raw materials in parts by mass:

[0010] 1 - 4 parts of EPA from microalgae oil, 1 - 2 parts of epigallocatechin - 3 - gallate, 0.5 - 0.8 parts of α - tocopherol, ginsenoside Rg 3 0.05 - 0.1 part, 2 - 10 parts of lipid, 1 - 10 parts of emulsifier, and water is made up to 100 parts.

[0011] In a possible embodiment, the EPA liposome is prepared and formed by self - assembly of liposomes with the following raw materials in parts by mass:

[0012] 4 parts of EPA from microalgae oil, 2 parts of epigallocatechin - 3 - gallate, 0.5 part of α - tocopherol, ginsenoside Rg 3 0.08 part, 5 parts of lipid, 6 parts of emulsifier, and water is made up to 100 parts.

[0013] In a possible embodiment, the lipid is at least one of butyl octyl salicylate, egg yolk lecithin, cholesterol, phosphatidylcholine, and phosphatidylserine.

[0014] In a possible embodiment, the lipid is one of the combination of egg yolk lecithin and cholesterol, and the combination of egg yolk lecithin and phosphatidylcholine.

[0015] In a possible embodiment, the emulsifier is at least one of PE40 hydrogenated castor oil, coconut glucoside, polyglyceryl - 10 stearate, PEG - 8 caprylic / capric glyceride, sucrose stearate, and polysorbate - 80.

[0016] In a possible embodiment, the emulsifier is the combination of polyglyceryl - 10 stearate and coconut glucoside.

[0017] The second aspect of the present invention also provides a method for preparing the EPA liposome described above, which includes:

[0018] Dissolve the lipid in ethanol and then evaporate it under reduced pressure to form a lipid film;

[0019] Add the emulsifier, EPA from microalgae oil, epigallocatechin - 3 - gallate, α - tocopherol, and ginsenoside Rg 3 into water and stir at 40 - 60 °C to form an aqueous phase;

[0020] Mix the aqueous phase with the lipid film at 40 - 60 °C and then perform high - speed shearing to obtain a primary liposome emulsion;

[0021] Perform high - pressure homogenization treatment on the primary liposome emulsion to obtain the EPA liposome.

[0022] In a possible embodiment, the pressure of the high - pressure homogenization treatment is 500 - 1000 bar, the temperature is 40 - 60 °C, and the number of homogenization times is 3 - 6.

[0023] In a possible implementation, the shearing rate is 8000 - 10000 rpm and the time is 1 - 5 min.

[0024] The third aspect of the present invention also provides the use of the above-mentioned EPA liposomes in the preparation of cosmetics.

[0025] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0026] The EPA liposomes provided by the present invention are prepared into nano-liposomes by setting the combination of active ingredients including algal oil EPA, epicatechin-3-gallate, α-tocopherol and ginsenoside Rg 3 so as to achieve the stable effect of EPA in composite systems such as cosmetics and skin care products through the synergy of the contained active ingredients and the encapsulation design of nano-liposomes, while enhancing the in-situ antioxidant and protection functions of EPA and slowing down the auto-oxidation of EPA, thereby improving the bioavailability and enhancing the anti-aging effect. Specifically, taking the contained algal oil EPA and epicatechin-3-gallate as the main active ingredients, supplemented by α-tocopherol and ginsenoside Rg 3 , on the one hand, it can synergistically with epicatechin-3-gallate to scavenge free radicals, inhibit antioxidant mechanisms such as oxidase, improve the antioxidant function and weaken the auto-oxidative degradation of EPA; on the other hand, it can synergistically with α-tocopherol to eliminate skin free radicals caused by ultraviolet light, reduce the oxidative damage caused by free radicals to the skin, achieve good photoprotection for the skin, and at the same time terminate the auto-oxidation of EPA by combining with the free radicals generated by the oxidation of EPA, effectively slowing down the oxidation degradation rate of EPA and improving the bioavailability of EPA; thirdly, it can synergistically with ginsenoside Rg 3 regulate the redox balance in the human body, enhance the self-repair of cells, reduce oxidative stress reactions, effectively enhance the in-situ antioxidant function of EPA, achieve a long-term anti-aging effect, and further expand the application prospects of EPA in the daily chemical industry. In addition, the present invention also improves the encapsulation rate of active ingredients by reasonably controlling the preparation parameters. Specifically, high-speed shearing can simply encapsulate the active ingredients, which can reduce the oxidation decomposition efficiency of the active ingredients when they are not homogenized into stable liposomes. By combining high-speed shearing and high-pressure homogenization, the uniformity of liposomes can be effectively guaranteed, and the particle size of liposomes can be regulated. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings required for the description of the embodiments will be briefly introduced below. Obviously, the drawings in the following description are only some embodiments recorded in the present application. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0028] Figure 1 The effect diagram of ABTS free radical scavenging rate provided for this application;

[0029] Figure 2 The transdermal absorption effect diagram provided for this application. Specific embodiments

[0030] Next, the technical solutions in the embodiments of this application will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of this application. Obviously, the described embodiments are part of the embodiments of this application, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments described in this application without creative efforts shall fall within the scope of protection of this application.

[0031] In the following description of this specification, the term "and / or" is used to describe the association relationship of associated objects, indicating that there can be three relationships. For example, A and / or B can represent: A exists alone, B exists alone, and both A and B exist simultaneously. Among them, A and B can be singular or plural; the symbol " / " means "or".

[0032] In the following description of this specification, the term "at least one" means one or more, and "multiple" means two or more. "At least one of the following" or similar expressions refer to any combination of these items, including any combination of single or plural items. For example, "at least one of A, B, or C", or, "at least one of A, B, and C", can represent any one of A, B, C, or A + B, or A + C, or B + C, or A + B + C, where A, B, and C can be single or multiple respectively.

[0033] In the following description of this specification, the sequence number does not mean the sequence of execution. Some or all steps can be executed in parallel or sequentially. The execution sequence of each process should be determined by its function and internal logic, and does not constitute any limitation to the execution process of this embodiment.

[0034] In the following description of this specification, the numerical range should be understood as also specifically disclosing each intermediate value between the upper and lower limits of the range. Any stated value or intermediate value within the stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in this embodiment, and the upper and lower limits of the smaller range can be independently included or excluded from the range.

[0035] Unless otherwise specified, the technical / scientific terms used in this specification have the meanings commonly understood by those of ordinary skill in the art. Although this specification only describes preferred materials and methods, any similar or equivalent methods and materials can also be used in specific examples or test cases. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the said documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0036] Example 1

[0037] This example provides a preparation method of EPA liposomes, specifically including:

[0038] S1: Add 3 parts by mass of egg yolk lecithin and 2 parts by mass of cholesterol to 10 parts by mass of ethanol solution. After forming a uniform lipid ethanol solution by magnetic stirring at 400 rpm and 50 °C, perform vacuum evaporation on the lipid ethanol solution to form a lipid film;

[0039] S2: Mix 4 parts by mass of polyglyceryl-10 stearate, 2 parts by mass of coco-glucoside, 4 parts by mass of microalgae oil EPA, 2 parts by mass of epicatechin-3-gallate, 0.5 part by mass of α-tocopherol, 0.08 part by mass of ginsenoside Rg 3 and the balance of water, and form a uniform solution by magnetic stirring at 300 rpm for 30 min at a temperature of 40 °C;

[0040] S3: Add the solution obtained in S2 to the lipid film obtained in S1, and form a uniform mixture by magnetic stirring at 500 rpm for 40 min at a temperature of 40 °C;

[0041] S4: Place the mixture obtained in S3 at room temperature and perform high-speed shearing at 9000 rpm for 10 min to obtain the primary liposome emulsion;

[0042] S5: Place the primary liposome emulsion obtained in S4 into a high-pressure homogenizer with a pressure of 600 bar and a temperature of 45 °C for homogenization 3 times, and cool to room temperature to obtain EPA liposomes.

[0043] Example 2

[0044] This example provides a preparation method of EPA liposomes, specifically including:

[0045] S1: Add 3 parts by mass of egg yolk lecithin and 2 parts by mass of phosphatidylcholine to 10 parts by mass of ethanol solution. After forming a uniform lipid ethanol solution by magnetic stirring at 400 rpm and 50 °C, perform vacuum evaporation on the lipid ethanol solution to form a lipid film;

[0046] S2: Mix 4 parts by mass of polyglyceryl-10 stearate, 2 parts by mass of cocoyl glucoside, 4 parts by mass of microalgae oil EPA, 2 parts by mass of epicatechin-3-gallate, 0.5 part by mass of α-tocopherol, 0.08 part by mass of ginsenoside Rg 3 with the balance of water, and magnetically stir at 300 rpm for 30 min at a temperature of 40 °C to form a homogeneous solution;

[0047] S3: Add the solution obtained in S2 to the lipid film obtained in S1, and magnetically stir at 500 rpm for 40 min at a temperature of 40 °C to form a homogeneous mixture;

[0048] S4: Place the mixture obtained in S3 at room temperature and subject it to high-speed shearing at 9000 rpm for 10 min to obtain the liposome primary emulsion;

[0049] S5: Place the liposome primary emulsion obtained in S4 into a high-pressure homogenizer with a pressure of 600 bar and a temperature of 45 °C for homogenization 3 times, and cool to room temperature to obtain the EPA liposome.

[0050] Example 3

[0051] This example provides the preparation of an EPA liposome, specifically including:

[0052] S1: Add 5 parts by mass of egg yolk lecithin to 10 parts by mass of an ethanol solution, magnetically stir at 400 rpm and form a homogeneous lipid ethanol solution at 50 °C, and then perform vacuum evaporation on the lipid ethanol solution to form a lipid film;

[0053] S2: Mix 4 parts by mass of polyglyceryl-10 stearate, 2 parts by mass of cocoyl glucoside, 4 parts by mass of microalgae oil EPA, 2 parts by mass of epicatechin-3-gallate, 0.5 part by mass of α-tocopherol, 0.08 part by mass of ginsenoside Rg 3 with the balance of water, and magnetically stir at 300 rpm for 30 min at a temperature of 40 °C to form a homogeneous solution;

[0054] S3: Add the solution obtained in S2 to the lipid film obtained in S1, and magnetically stir at 500 rpm for 40 min at a temperature of 40 °C to form a homogeneous mixture;

[0055] S4: Place the mixture obtained in S3 at room temperature and subject it to high-speed shearing at 9000 rpm for 10 min to obtain the liposome primary emulsion;

[0056] S5: Place the liposome primary emulsion obtained in S4 into a high-pressure homogenizer with a pressure of 600 bar and a temperature of 45 °C for homogenization 3 times, and cool to room temperature to obtain the EPA liposome.

[0057] Example 4

[0058] This embodiment provides a method for preparing EPA liposomes, specifically including:

[0059] S1: Add 3 parts by mass of egg yolk lecithin and 2 parts by mass of cholesterol to 10 parts by mass of ethanol solution. After forming a uniform lipid ethanol solution through magnetic stirring at 400 rpm and 50 °C, perform vacuum evaporation on the lipid ethanol solution to form a lipid film;

[0060] S2: Mix 4 parts by mass of polyglyceryl-10 stearate, 2 parts by mass of coco-glucoside, 4 parts by mass of microalgae oil EPA, 2 parts by mass of epicatechin-3-gallate, 0.8 parts by mass of α-tocopherol, 0.1 parts by mass of ginsenoside Rg 3 and the balance of water, and form a uniform solution through magnetic stirring at 300 rpm for 30 min at a temperature of 40 °C;

[0061] S3: Add the solution obtained in S2 to the lipid film obtained in S1, and form a uniform mixture through magnetic stirring at 500 rpm for 40 min at a temperature of 40 °C;

[0062] S4: Place the mixture obtained in S3 at room temperature and perform high-speed shearing at 9000 rpm for 10 min to obtain the liposome primary emulsion;

[0063] S5: Place the liposome primary emulsion obtained in S4 into a high-pressure homogenizer with a pressure of 600 bar and a temperature of 45 °C for homogenization 3 times, and cool to room temperature to obtain EPA liposomes.

[0064] Example 5

[0065] This embodiment provides a method for preparing EPA liposomes, specifically including:

[0066] S1: Add 3 parts by mass of egg yolk lecithin and 2 parts by mass of cholesterol to 10 parts by mass of ethanol solution. After forming a uniform lipid ethanol solution through magnetic stirring at 400 rpm and 50 °C, perform vacuum evaporation on the lipid ethanol solution to form a lipid film;

[0067] S2: Mix 4 parts by mass of polyglyceryl-10 stearate, 2 parts by mass of coco-glucoside, 4 parts by mass of microalgae oil EPA, 2 parts by mass of epicatechin-3-gallate, 0.2 parts by mass of α-tocopherol, 0.02 parts by mass of ginsenoside Rg 3 and the balance of water, and form a uniform solution through magnetic stirring at 300 rpm for 30 min at a temperature of 40 °C;

[0068] S3: Add the solution obtained in S2 to the lipid film obtained in S1, and form a uniform mixture through magnetic stirring at 500 rpm for 40 min at a temperature of 40 °C;

[0069] S4: Place the mixture obtained in S3 at room temperature and shear it at a high speed of 9000 rpm for 10 min to obtain the liposome primary emulsion;

[0070] S5: Place the liposome primary emulsion obtained in S4 into a high-pressure homogenizer with a pressure of 600 bar and a temperature of 45 °C for homogenization 3 times, and then cool it to room temperature to obtain EPA liposomes.

[0071] Example 6

[0072] This example provides a preparation method of EPA liposomes, which specifically includes:

[0073] S1: Add 3 parts by mass of egg yolk lecithin and 2 parts by mass of cholesterol to 10 parts by mass of an ethanol solution. After magnetic stirring at 400 rpm and 50 °C to form a uniform lipid ethanol solution, perform vacuum evaporation on the lipid ethanol solution to form a lipid film;

[0074] S2: Mix 4 parts by mass of polyglyceryl-10 stearate, 2 parts by mass of coco-glucoside, 4 parts by mass of microalgae oil EPA, 2 parts by mass of epigallocatechin-3-gallate, 1 part by mass of α-tocopherol, 0.1 part by mass of ginsenoside Rg 3 and the balance of water, and stir magnetically at 300 rpm for 30 min at a temperature of 40 °C to form a uniform solution;

[0075] S3: Add the solution obtained in S2 to the lipid film obtained in S1, and stir magnetically at 500 rpm for 40 min at a temperature of 40 °C to form a uniform mixture;

[0076] S4: Place the mixture obtained in S3 at room temperature and shear it at a high speed of 9000 rpm for 10 min to obtain the liposome primary emulsion;

[0077] S5: Place the liposome primary emulsion obtained in S4 into a high-pressure homogenizer with a pressure of 600 bar and a temperature of 45 °C for homogenization 3 times, and then cool it to room temperature to obtain EPA liposomes.

[0078] Comparative Example 1

[0079] This comparative example provides a preparation method of EPA liposomes, which specifically includes:

[0080] S1: Add 3 parts by mass of egg yolk lecithin and 2 parts by mass of cholesterol to 10 parts by mass of an ethanol solution. After magnetic stirring at 400 rpm and 50 °C to form a uniform lipid ethanol solution, perform vacuum evaporation on the lipid ethanol solution to form a lipid film;

[0081] S2: Mix 4 parts by mass of polyglyceryl-10 stearate, 2 parts by mass of cocoglucoside, 4 parts by mass of microalgal oil EPA, and the balance of water, and magnetically stir at 300 rpm for 30 min at a temperature of 40 °C to form a homogeneous solution;

[0082] S3: Add the solution obtained in S2 to the lipid film obtained in S1, and magnetically stir at 500 rpm for 40 min at a temperature of 40 °C to form a homogeneous mixture;

[0083] S4: Place the mixture obtained in S3 at room temperature and subject it to high-speed shearing at 9000 rpm for 10 min to obtain the liposome primary emulsion;

[0084] S5: Homogenize the liposome primary emulsion obtained in S4 three times in a high-pressure homogenizer at a pressure of 600 bar and a temperature of 45 °C, and cool to room temperature to obtain the EPA liposome.

[0085] Comparative Example 2

[0086] This comparative example provides a preparation of EPA liposome, specifically including:

[0087] S1: Add 3 parts by mass of egg yolk lecithin and 2 parts by mass of cholesterol to 10 parts by mass of ethanol solution. After magnetically stirring at 400 rpm and forming a homogeneous lipid ethanol solution at 50 °C, perform reduced-pressure evaporation on the lipid ethanol solution to form a lipid film;

[0088] S2: Mix 4 parts by mass of polyglyceryl-10 stearate, 2 parts by mass of cocoglucoside, 4 parts by mass of microalgal oil EPA, 0.5 part by mass of α-tocopherol, 0.08 part by mass of ginsenoside Rg 3 and the balance of water, and magnetically stir at 300 rpm for 30 min at a temperature of 40 °C to form a homogeneous solution;

[0089] S3: Add the solution obtained in S2 to the lipid film obtained in S1, and magnetically stir at 500 rpm for 40 min at a temperature of 40 °C to form a homogeneous mixture;

[0090] S4: Place the mixture obtained in S3 at room temperature and subject it to high-speed shearing at 9000 rpm for 10 min to obtain the liposome primary emulsion;

[0091] S5: Homogenize the liposome primary emulsion obtained in S4 three times in a high-pressure homogenizer at a pressure of 600 bar and a temperature of 45 °C, and cool to room temperature to obtain the EPA liposome.

[0092] Comparative Example 3

[0093] This comparative example provides a preparation of EPA liposome, specifically including:

[0094] S1: Add 3 parts by mass of egg yolk lecithin and 2 parts by mass of cholesterol into 10 parts by mass of ethanol solution. After forming a uniform lipid ethanol solution through magnetic stirring at 400 rpm and at 50 °C, perform reduced-pressure evaporation on the lipid ethanol solution to form a lipid film;

[0095] S2: Mix 4 parts by mass of polyglyceryl-10 stearate, 2 parts by mass of cocoyl glucoside, 4 parts by mass of microalgae oil EPA, 2 parts by mass of epicatechin-3-gallate and the balance of water, and form a uniform solution through magnetic stirring at 300 rpm for 30 min at a temperature of 40 °C;

[0096] S3: Add the solution obtained in S2 into the lipid film obtained in S1, and form a uniform mixture through magnetic stirring at 500 rpm for 40 min at a temperature of 40 °C;

[0097] S4: Place the mixture obtained in S3 at room temperature and perform high-speed shearing at 9000 rpm for 10 min to obtain the primary lipid emulsion;

[0098] S5: Place the primary lipid emulsion obtained in S4 into a high-pressure homogenizer with a pressure of 600 bar and a temperature of 45 °C for homogenization 3 times, and cool to room temperature to obtain the EPA liposome.

[0099] Comparative Example 4

[0100] This comparative example provides the preparation of an EPA liposome, specifically including:

[0101] S1: Add 3 parts by mass of egg yolk lecithin and 2 parts by mass of cholesterol into 10 parts by mass of ethanol solution. After forming a uniform lipid ethanol solution through magnetic stirring at 400 rpm and at 50 °C, perform reduced-pressure evaporation on the lipid ethanol solution to form a lipid film;

[0102] S2: Mix 4 parts by mass of polyglyceryl-10 stearate, 2 parts by mass of cocoyl glucoside, 4 parts by mass of microalgae oil EPA, 2 parts by mass of epicatechin-3-gallate, 0.5 part by mass of α-tocopherol and the balance of water, and form a uniform solution through magnetic stirring at 300 rpm for 30 min at a temperature of 40 °C;

[0103] S3: Add the solution obtained in S2 into the lipid film obtained in S1, and form a uniform mixture through magnetic stirring at 500 rpm for 40 min at a temperature of 40 °C;

[0104] S4: Place the mixture obtained in S3 at room temperature and perform high-speed shearing at 9000 rpm for 10 min to obtain the primary lipid emulsion;

[0105] S5: Place the primary lipid emulsion obtained in S4 into a high-pressure homogenizer with a pressure of 600 bar and a temperature of 45 °C for homogenization 3 times, and cool to room temperature to obtain the EPA liposome.

[0106] Comparative Example 5

[0107] This comparative example provides a preparation method of EPA liposomes, specifically including:

[0108] S1: Add 3 parts by mass of egg yolk lecithin and 2 parts by mass of cholesterol into 10 parts by mass of ethanol solution. After forming a uniform lipid ethanol solution through magnetic stirring at 400 rpm and at 50 °C, perform reduced-pressure evaporation on the lipid ethanol solution to form a lipid film;

[0109] S2: Mix 4 parts by mass of polyglyceryl-10 stearate, 2 parts by mass of cocoyl glucoside, 4 parts by mass of microalgae oil EPA, 2 parts by mass of epicatechin-3-gallate, 0.08 part by mass of ginsenoside Rg 3 and the balance of water, and form a uniform solution through magnetic stirring at 300 rpm for 30 min at a temperature of 40 °C;

[0110] S3: Add the solution obtained in S2 to the lipid film obtained in S1, and form a uniform mixture through magnetic stirring at 500 rpm for 40 min at a temperature of 40 °C;

[0111] S4: Place the mixture obtained in S3 at room temperature and perform high-speed shearing at 9000 rpm for 10 min to obtain the primary liposome emulsion;

[0112] S5: Place the primary liposome emulsion obtained in S4 into a high-pressure homogenizer with a pressure of 600 bar and a temperature of 45 °C for homogenization 3 times, and cool to room temperature to obtain EPA liposomes.

[0113] 1. Particle size stability test

[0114] Perform particle size stability tests on the samples prepared in Examples 1-6 and Comparative Examples 1-5. Place them at -18 °C, 4 °C, room temperature, and 40 °C for 30 days. At the initial state, after being placed at -18 °C, 4 °C, room temperature, and 40 °C for 30 days, use a Zetasizer / Nano-ZS90 Malvern nano-potential particle size analyzer to measure the particle size under different temperature conditions at room temperature. The results are shown in Table 1.

[0115] Table 1: Results of particle size stability test of EPA liposomes

[0116]

[0117]

[0118] As can be seen from Table 1, the particle sizes of the liposomes prepared in Examples 1-6 and Comparative Examples 1-5 are all small, and the change range of the particle size at the storage temperature is not large. Among them, the particle size of the sample in Example 1 is the smallest, and the particle size is the most stable under the four storage temperatures.

[0119] 2. Entrapment efficiency test

[0120] The liposome samples prepared in Examples 1, 2, and 3 were processed, and the EPA content was analyzed by a Shimadzu gas chromatograph GC-2010Plus, and the entrapment efficiency of the samples was calculated.

[0121] Table 2: Results of entrapment efficiency of EPA liposome samples

[0122] Group Entrapment efficiency (%) Example 1 99.3 Example 2 97.5 Example 3 92.2

[0123] As can be seen from Table 2, the emulsification entrapment efficiencies of Examples 1-2 are relatively close and significantly higher than that of Example 3, indicating that the combination of egg yolk lecithin and cholesterol or phosphatidylcholine can improve the entrapment efficiency; at the same time, the emulsification entrapment efficiency of Example 1 is the highest, indicating that the combination of egg yolk lecithin and cholesterol has the best entrapment effect. Therefore, the present invention preferably uses a combined lipid of egg yolk lecithin and cholesterol or phosphatidylcholine, and more preferably a combined lipid of egg yolk lecithin and cholesterol.

[0124] 3. Antioxidant experiment

[0125] Mix 10 μL of the test substance evenly with 190 μL of ABTS working solution / PBS with a concentration of 0.25 mmol / L, shake well, to obtain the ABTS working solution solution of the test substance and the PBS solution of the test substance, incubate at room temperature for 6 min, and measure the absorbance at 405 nm. The results are Figure 1 as shown in Table 3. Among them, the absorbance of the ABTS working solution solution of the test substance is denoted as A1, the absorbance of the PBS solution of the test substance is denoted as A2, and the absorbance of the blank ABTS working solution and the blank PBS is denoted as A3; the test substance is the algal oil EPA liposome prepared in the examples and comparative examples. The calculation formula for the ABTS radical scavenging rate is as follows:

[0126]

[0127] Table 3: Results of ABTS radical scavenging rate

[0128]

[0129] According to Figure 1 and Table 3, the highest average value of the ABTS radical scavenging rate of the liposome emulsion prepared in Example 1 is 94.02%, and the ABTS radical scavenging rates of the examples are all higher than those of the comparative examples. All the examples of the invention can scavenge ABTS radicals and have antioxidant properties.

[0130] 4. Test the degradation rate of EPA in Examples 1-6 and Comparative Examples 1-5

[0131] The samples prepared in Examples 1-6 and Comparative Examples 1-5 were placed at room temperature for 30 days. At 0 day, 7 days, 14 days, 21 days, and 30 days, the samples were pretreated respectively. Then, a Shimadzu LC-20A liquid chromatograph was used to separate EPA and its degradation products, and quantitative analysis was carried out by an ultraviolet detector to calculate the EPA degradation rate. The results are shown in Table 4.

[0132] Table 4: Degradation rate of EPA

[0133]

[0134] According to Table 4, from the test data of the EPA degradation rate of the samples prepared in Examples 1-6 and Comparative Examples 1-5, it can be obtained that the EPA degradation rate of the samples prepared in Examples 1-6 is relatively slow, and the EPA degradation rate of the sample in Example 1 is the smallest.

[0135] 5. Transdermal absorption experiment

[0136] In vitro skin penetration experiment: The Franz diffusion cell was placed in a drug transdermal diffusion tester, and the condition was set as a constant temperature circulating water bath at 37 °C. The mouse skin was laid flat between the supply pool and the receiving pool and fixed with a specific clip. The stratum corneum of the skin faced the supply pool, and the subcutaneous tissue faced the receiving pool. The receiving pool was filled with 6 mL of receiving solution (PBS with pH 7.2-7.4), and air bubbles were removed. It was balanced at 37 °C and 300 r / min for 1 h. 1 mL of the sample to be tested was added to the supply pool respectively, and the preparation was contacted with the intact components of the stratum corneum. The supply pool was sealed with a sealing film to prevent the preparation from volatilizing, and the receiving solution was continuously stirred to ensure the uniform dispersion of the preparation; 1 mL of the receiving solution was taken out at 0, 2, 4, 8, and 24 h respectively, and an equal amount of fresh receiving solution PBS was immediately added for supplementation after each sampling. The taken samples were determined by HPLC and the cumulative permeability per unit skin area was calculated. The specific test results are as follows:

[0137] Cumulative permeation amount (Q) = (A × V) / A; V is the volume of the receiving chamber (milliliter); A is the exposed area of the skin sample (square centimeter).

[0138] Cumulative permeability (J): J = Q / (A × t), where Q is the amount of drug permeating through the skin within time t; A is the exposed area of the skin sample (square centimeter); t is the experimental time (hour).

[0139] According to Figure 2 it can be known that after a 24-hour penetration experiment, the cumulative permeation amount and permeability of all samples through the mouse skin increased with the extension of time. After 24 h, the cumulative transdermal permeability of Example 1 reached 5.51%, and the skin penetration effect was the best.

[0140] The various embodiments in this specification are described in a progressive manner. For the same or similar parts between the various embodiments, reference can be made to each other. Each embodiment focuses on the differences from other embodiments.

[0141] The above embodiments are only used to illustrate the technical solutions of the present application, rather than limiting the present application. Although the present application has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the present application.

Claims

1. An EPA liposome, characterized in that The liposomes are prepared by self-assembly of the following raw materials in parts by weight: Microalgae oil EPA 1-4 parts, epicatechin-3-gallate 1-2 parts, α-tocopherol 0.2-1 parts, ginsenoside Rg3 0.02-0.1 parts, lipid 2-10 parts, emulsifier 1-10 parts, water is added to 100 parts.

2. The EPA liposome according to claim 1, characterized in that The liposomes are prepared by self-assembly of the following raw materials in parts by weight: Microalgae oil EPA 1-4 parts, epicatechin-3-gallate 1-2 parts, α-tocopherol 0.5-0.8 parts, ginsenoside Rg3 0.05-0.1 parts, lipid 2-10 parts, emulsifier 1-10 parts, water is added to make up to 100 parts.

3. The EPA liposome according to claim 2, characterized in that The liposomes are prepared by self-assembly of the following raw materials in parts by weight: EPA 4 parts of microalgae oil, epicatechin-3-gallate 2 parts, α-tocopherol 0.5 parts, ginsenoside Rg30.08 parts, lipid 5 parts, emulsifier 6 parts, water is added to 100 parts.

4. The EPA liposome according to any one of claims 1 to 3, characterized in that The lipid is at least one of butyl octanol salicylate, egg yolk lecithin, cholesterol, phosphatidylcholine, and phosphatidylserine.

5. The EPA liposome according to claim 4, characterized in that The lipid is one of a combination of egg yolk lecithin and cholesterol and a combination of egg yolk lecithin and phosphatidylcholine.

6. The EPA liposome according to any one of claims 1 to 3, characterized in that The emulsifier is at least one of PE40 hydrogenated castor oil, coco-glucoside, polyglyceryl-10 stearate, PEG-8 caprylic / capric glyceride, sucrose stearate, and polysorbate-80.

7. The EPA liposome according to claim 6, characterized in that The emulsifier is a combination of polyglyceryl-10 stearate and coconut glucoside.

8. A method for preparing the EPA liposome according to any one of claims 1 to 7, characterized in that: Include: After dissolving the lipids in ethanol, the lipids are evaporated under reduced pressure to form a lipid film; Add emulsifier, microalgae oil EPA, epicatechin-3-gallate, α-tocopherol and ginsenoside Rg3 into water, and stir at 40-60° C. to form an aqueous phase; The aqueous phase and the lipid film are mixed at 40-60° C. and sheared at high speed to obtain liposome colostrum; The liposome colostrum is subjected to high pressure homogenization to obtain the EPA liposome.

9. The preparation method according to claim 8, characterized in that: The pressure of the high-pressure homogenization treatment is 500-1000 bar, the temperature is 40-60° C., and the number of homogenization times is 3-6; And / or, the shearing rate is 8000-10000 rpm, and the time is 1-5 min.

10. Use of the EPA liposome according to any one of claims 1 to 7 in preparing cosmetics.

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