A liposome and a method for preparing the same

Abstract

The application provides a kind of liposome, including phospholipid bilayer vesicular structure, the composition of the phospholipid bilayer vesicular structure includes phospholipid and long chain polyunsaturated fatty acid oil;The mass ratio of the phospholipid and the long chain polyunsaturated fatty acid oil is 1:(1-4);The long chain polyunsaturated fatty acid oil is DHA oil;The content of DHA in the DHA oil is 35% to 70%.The liposome provided in the application contains DHA and other long chain polyunsaturated fatty acids, DHA and other long chain polyunsaturated fatty acids as the composition of the phospholipid bilayer vesicular structure, so that the phase transition temperature of the liposome is increased, the flowability is weakened, and the structure is more stable;The surface of the liposome is charged, and the suspension stability is good.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of nanocarrier transport, in particular to a liposome and a preparation method thereof. BACKGROUND

[0002] Liposomes are spherical vesicles with a bilayer structure composed of amphiphilic substances-phospholipids, which are composed of single or multi-chamber phospholipid bilayer vesicles. Liposomes have the characteristics of biocompatibility and sustained release, and are a promising carrier delivery system.

[0003] As the main component of liposomes, the quality of phospholipids directly affects the quality of liposomes produced. Phospholipids include synthetic phospholipids and natural phospholipids. Synthetic phospholipids mainly include DPPC (dipalmitoyl phosphatidylcholine), DPPE (dipalmitoyl phosphatidyl ethanolamine), DSPC (distearoyl phosphatidylcholine) and the like, which all belong to hydrogenated phospholipids, and have the characteristics of high purity, stable properties, strong antioxidant properties, and stable finished products. However, synthetic phospholipids are expensive, and a large amount of organic solvent is used in production, which may pose a safety hazard. Natural phospholipids are mainly egg phospholipids (phosphatidylcholine, PC), and the main sources include egg yolk and soybeans, which are neutral. Due to the complex and diverse sources, types and compositions of natural phospholipids, they are a mixture, and the types of hydrophilic groups, the length of fatty acid chains and the degree of saturation are very different, the purity is difficult to control, and there are large differences between batches, and the stability is poor during use and storage; different phospholipids exhibit different physicochemical properties, which also affect the physicochemical property indicators and stability of liposomes, and even affect the functional properties of liposomes. Currently, high-purity natural phospholipids on the market also have problems such as high price and organic solvent residue.

[0004] Docosahexaenoic acid (DHA) has a molecular formula of C 22 H 32 O2, which is a straight-chain fatty acid containing 22 carbon atoms and 6 double bonds. DHA is a polyunsaturated fatty acid that is very important to the human body, and is an important member of the Omega-3 unsaturated fatty acid family. DHA is a major element for the growth and maintenance of nervous system cells, and is an important constituent fatty acid of the brain and retina.

[0005] Current technologies for DHA in liposomes include:

[0006] Patent CN105231460A, the mass ratio of soybean phospholipid to phospholipid type DHA is 10:1-4, Tween is an emulsifying agent, and DHA liposomes are obtained by ultrasonic treatment after hydration and membrane washing;

[0007] Patent CN113331423A, the mass ratio of phospholipid to DHA algal oil is 4-10:1, and octenyl succinate starch sodium is used as an emulsifier stabilizing system;

[0008] Patent CN109589321B, the mass ratio of phospholipid to DHA algal oil is 2:1, and a gas-assisted jet reaction chamber is designed to prepare DHA liposomes;

[0009] Patent CN103230002A, the mass ratio of DHA to lecithin is 0.5-1:2-6, and DHA liposomes are prepared by traditional film method combined with new dynamic high pressure method;

[0010] In the literature "Preparation and properties of DHA liposomes", the mass ratio of DHA:soy lecithin to cholesterol is 1:10.

[0011] In the literature "Preparation and properties of DHA algal oil liposomes", the mass ratio of soy lecithin to DHA algal oil is 5:1, and the encapsulation efficiency and particle size of the prepared liposomes have good indicators.

[0012] Based on the above-mentioned public literature, it can be seen that the main application of DHA liposomes at present is to encapsulate DHA as an active substance inside the liposomes to enhance the biocompatibility and tissue utilization of DHA. There is no report in the prior art that DHA is used as part of the liposome encapsulating material to improve the stability, encapsulation efficiency and other related qualities of the liposomes. SUMMARY

[0013] In view of the problems existing in the prior art, the present application provides a liposome containing DHA oil and a preparation method thereof. The addition of the DHA oil can significantly improve the stability of the liposome during preparation, storage and transportation; at the same time, it can improve the encapsulation efficiency of the liposome for some active ingredients, and can overcome the adverse effects of complex natural phospholipids on the physicochemical property indexes and stability of the liposome.

[0014] The present application provides a liposome, which comprises a phospholipid bilayer vesicular structure, and the composition of the phospholipid bilayer vesicular structure comprises phospholipid and long-chain polyunsaturated fatty acid oil; the mass ratio of the phospholipid to the long-chain polyunsaturated fatty acid oil is 1:(1-4);

[0015] The long-chain polyunsaturated fatty acid oil is DHA oil; the content of DHA in the DHA oil is 35%-70%. It is recognized by those skilled in the art that the content of DHA is obtained by internal standard method to obtain the mass fraction of DHA in total oil.

[0016] In the present application, DHA and other long-chain polyunsaturated fatty acids are used as a component of the phospholipid bilayer vesicle structure (i.e. part of the encapsulating material, rather than an active substance to be encapsulated), which is loaded by non-covalent interaction accumulation between the unsaturated fatty acid functional groups in the phospholipid structure, enhancing the hydrophobicity of the liposome vesicle lipophilic group, promoting the interaction of the phospholipid bilayer, increasing the thickness of the phospholipid bilayer; increasing the phospholipid phase transition temperature, reducing the fluidity, and further stabilizing the liposome structure. At the same time, DHA and other long-chain polyunsaturated fatty acids also enhance the binding of the phospholipid structure and the poorly soluble active ingredient, and improve the encapsulation efficiency of the liposome for the poorly soluble active ingredient. In addition, the carboxyl group of the long-chain polyunsaturated fatty acid in DHA oil is embedded in the phospholipid hydrophilic group, providing a negative charge for the neutral phospholipid, so that the liposome itself carries a charge, and there is an electrostatic repulsion between the liposomes, so the stability of the liposome suspension can be maintained and further improved.

[0017] The prior art patent CN108741080B shows that soybean phospholipid 4-10 mg / mL, microalgae DHA 2-5 mg / mL, and anthocyanin 1.33-3.33 mg / mL constitute a DHA-anthocyanin double-phase nanoliposome; although the ratio of phospholipid and DHA oil similar to the present application appears in the range, the document only lists the choice of 2:1 in the examples, and for this choice, it does not disclose the use of DHA oil to improve the stability of the liposome as described in the present application, nor does it give how to determine the ratio to overcome the influence of the complex natural phospholipid on the physicochemical property indexes and stability of the liposome.

[0018] Preferably, the mass ratio of the phospholipid to the long-chain polyunsaturated fatty acid oil is 1:(1-2).

[0019] Preferably, the content of triglyceride in the long-chain polyunsaturated fatty acid oil is greater than 95%.

[0020] Preferably, the phospholipid includes soybean lecithin and / or egg yolk lecithin.

[0021] Preferably, the particle size of the liposome is 80-180 nm.

[0022] Preferably, the DHA oil further includes one or more of ALA, EPA, LA, AA, and DPA; the total mass proportion of the polyunsaturated fatty acid oil in the DHA oil is not less than 50%.

[0023] Preferably, the liposome further includes a co-surfactant and / or a surfactant.

[0024] The co-surfactant includes a polyol and / or a polyethylene glycol.

[0025] The polyhydric alcohols include one or more of propylene glycol, butylene glycol and glycerol;

[0026] The polyethylene glycols include one or more of polyethylene glycol 400, polyethylene glycol 1000 and polyethylene glycol 2000;

[0027] The surfactants include one or more of Tween 20, Tween 40, Tween 60, Tween 80, PEG-10 hydrogenated castor oil, PEG-20 hydrogenated castor oil, PEG-30 hydrogenated castor oil, PEG-40 hydrogenated castor oil, PEG-50 hydrogenated castor oil and PEG-60 hydrogenated castor oil.

[0028] Preferably, the liposomes further include an encapsulated active ingredient; the encapsulated active ingredient includes one or more of vitamin A, vitamin A ester, vitamin E, vitamin E ester, vitamin D2, vitamin D3, vitamin K, biotin, coenzyme Q10, curcumin, beta-carotene, lutein, canthaxanthin, lutein ester, lycopene and astaxanthin.

[0029] Preferably, the mass ratio of the phospholipid, the long-chain polyunsaturated fatty acid oil and the encapsulated active ingredient is 1:(1-4):(0.1-2).

[0030] Preferably, the mass ratio of the phospholipid, the long-chain polyunsaturated fatty acid oil and the encapsulated active ingredient is 1:(1-2):(0.1-2).

[0031] The present application provides a method for preparing the liposomes, including the following steps:

[0032] S1, weighing the liposome components with the mass ratio; the liposome components include the phospholipid and the long-chain polyunsaturated fatty acid oil; or, the liposome components include the phospholipid, the long-chain polyunsaturated fatty acid oil and an encapsulated active ingredient;

[0033] S2, mixing the liposome components in step S1 with anhydrous ethanol to obtain a mixed solution;

[0034] S3, evaporating the alcohol in the mixed solution in step S2 to obtain an oily film;

[0035] S4, placing the oily film in step S3 in a dispersion medium or a dispersion medium containing the surfactant to obtain a liposome dispersion; the dispersion medium includes water and / or the co-surfactant.

[0036] The present application further provides another kind of liposomes, which include the liposome dispersion in step S4 and a fragrance; the mass proportion of the fragrance in the liposomes is 0.001%-0.1%.

[0037] Advantages:

[0038] The application forms a bilayer vesicular structure of liposome by specific proportion of DHA oil and phospholipid, the addition of DHA oil rich in polyunsaturated fatty acid can significantly improve the stability of the liposome in the process of preparation, storage and transportation; meanwhile, it can improve the encapsulation rate of the liposome to some active ingredients, and overcome the adverse effects of complex natural phospholipid on the physical and chemical property indexes and stability of the liposome, so that the selection of phospholipid as one of the components is more extensive. DETAILED DESCRIPTION

[0039] The application provides a liposome, which comprises a phospholipid bilayer vesicular structure, and the composition of the phospholipid bilayer vesicular structure comprises phospholipid and long-chain polyunsaturated fatty acid oil.

[0040] In the application, the phospholipid comprises natural phospholipid and / or synthetic phospholipid, and the natural phospholipid preferably comprises soybean lecithin and / or egg yolk lecithin. One of the advantages of the application is that the influence of complex natural phospholipid on the physical and chemical property indexes and stability of the liposome can be overcome, so that the purity and source of the phospholipid are not specially required, and the conventional commercially available phospholipid products can be used as the raw material for preparing the liposome.

[0041] In the application, the long-chain polyunsaturated fatty acid oil is DHA oil; the DHA oil comprises DHA; the mass ratio of the DHA in the DHA oil is 35%-70%; the DHA oil preferably further comprises one or more of α-linolenic acid (ALA), eicosapentaenoic acid (EPA), linoleic acid (LA), arachidonic acid (AA) and docosapentaenoic acid (DPA); more preferably, the DHA oil comprises EPA, DPA and AA, wherein the content of the EPA is preferably 1-10%, the content of the DPA is preferably 8-20%, and the content of the AA is preferably 1-5%. As a preferred scheme, in the DHA oil, the mass ratio of the total polyunsaturated fatty acid oil in the DHA oil is not less than 50%. In the field, the long-chain polyunsaturated fatty acid is understood as a polyunsaturated fatty acid with more than 20 carbon atoms and more than 2 unsaturated double bonds.

[0042] In the present application, the long-chain polyunsaturated fatty acid in the DHA oil preferably includes one or more of free type, ethyl ester type, triglyceride type and phospholipid type; more preferably includes triglyceride type. In a more preferred embodiment of the present application, the content of triglyceride in the long-chain polyunsaturated fatty acid oil is greater than 95%. The source of the DHA oil in the present application is not particularly limited, and any DHA oil that is commonly commercially available in the art and within the required specification range of the present application can be used.

[0043] In the present application, the mass ratio of the phospholipid to the long-chain polyunsaturated fatty acid oil is 1:(1-4), preferably 1:(1-2). The liposome with the mass ratio within the range has good stability. If the ratio is lower than this, the stability cannot be achieved due to the too low content of phospholipid.

[0044] In the present application, the bilayer vesicle of the liposome is preferably large unilamellar vesicle. The particle size of the liposome is preferably 80-180 nm, and the stability is better.

[0045] The liposome provided by the present application has the characteristics of uniform particle size, negative surface charge and strong antioxidant capacity. The addition of long-chain polyunsaturated fatty acid oil in the raw material can enhance the stability of the liposome and improve the encapsulation rate of the poorly soluble active ingredient. The influence of natural phospholipid with complex components on the physicochemical property index and stability of the liposome can be effectively overcome.

[0046] In the present application, the liposome preferably further includes a co-surfactant and / or a surfactant; the co-surfactant preferably includes one or more of polyhydric alcohol and / or polyethylene glycol; the polyhydric alcohol preferably includes one or more of propylene glycol, butylene glycol and glycerol; the polyethylene glycol preferably includes one or more of polyethylene glycol 400, polyethylene glycol 1000 and polyethylene glycol 2000; the surfactant preferably includes one or more of Tween 20, Tween 40, Tween 60, Tween 80, PEG-10 hydrogenated castor oil, PEG-20 hydrogenated castor oil, PEG-30 hydrogenated castor oil, PEG-40 hydrogenated castor oil, PEG-50 hydrogenated castor oil and PEG-60 hydrogenated castor oil. In the present application, the total mass of the co-surfactant and / or surfactant and the mass of the phospholipid and the long-chain polyunsaturated fatty acid preferably have a mass ratio of (0-2):1:(1-4), more preferably (0-2):1:(1-2). The source of the co-surfactant and / or surfactant in the present application is not particularly limited, and any product that is commonly commercially available in the art and meets the standard can be used.

[0047] In the present application, the liposome preferably further comprises an entrapped active ingredient; the entrapped active ingredient preferably comprises one or more of vitamin A, vitamin A ester, vitamin E, vitamin E ester, vitamin D2, vitamin D3, vitamin K, biotin, coenzyme Q10, curcumin, beta-carotene, lutein, canthaxanthin, lutein ester, lycopene and astaxanthin; more preferably comprises one or more of vitamin A, coenzyme Q10, beta-carotene and lutein ester. In the present application, the mass ratio of the phospholipid, the long-chain polyunsaturated fatty acid oil and the entrapped active ingredient is preferably 1:(1-4):(0.1-2), more preferably 1:(1-2):(0.1-2). The present application does not make special limitations on the source of the entrapped active ingredient, and those skilled in the art can select from the conventional commercially available products meeting the standards according to actual needs.

[0048] The present application provides a preparation method of the liposome, comprising the following steps:

[0049] S1, weighing the liposome components with the mass ratio; the liposome components comprise the phospholipid and the long-chain polyunsaturated fatty acid oil; or, the components of the liposome comprise the phospholipid, the long-chain polyunsaturated fatty acid oil and the entrapped active ingredient;

[0050] S2, dissolving the liposome components in anhydrous ethanol according to step S1;

[0051] S3, evaporating the ethanol solution obtained by dissolving according to step S2 to obtain an oily film;

[0052] S4, placing the oily film according to step S3 in a dispersion medium or a dispersion medium containing the surfactant to obtain a liposome dispersion; the dispersion medium comprises water and / or the co-surfactant.

[0053] The preparation method provided by the present application adds long-chain polyunsaturated fatty acid as an oil phase in the process of preparing liposomes from phospholipids, uses the phospholipid bilayer vesicles formed by the combination of long-chain polyunsaturated fatty acid and phospholipid, obtains long-chain polyunsaturated fatty acid liposomes with uniform particle size, negative surface charge, strong antioxidant capacity, improved encapsulation rate of active ingredients, and can enhance the stability of the liposomes.

[0054] The present application also provides another kind of liposome, which comprises the liposome dispersion according to step S4 and a fragrance; the mass ratio of the fragrance in the liposome is 0.001%-0.1%. The present application does not make special limitations on the type and source of the fragrance, and the conventional commercially available products meeting the standards can be used.

[0055] The liposome can be obtained by using the high-pressure homogenization method, and industrial preparation is realized.

[0056] The liposome provided by the application has simple preparation steps, low preparation condition requirement, good product quality and high stability, and provides various optional paths for dry preparation of the liposome.

[0057] The technical solutions provided by the application will be described in detail below in combination with examples, but they should not be understood as limiting the protection scope of the application. If not specifically indicated, the experimental methods used in the examples are conventional methods; the materials, reagents and the like used can be obtained from commercial channels.

[0058] In each example, different types of soybean lecithin and ethanol are purchased from the National Medicine. The DHA oil is DHA algal oil produced by Ji Biyou and sold in the market, and the DHA content is 45%. The contents of lutein ester, beta-carotene and astaxanthin are all above 95%. Vitamin A and coenzyme Q10 are purchased from Jin Dawei.

[0059] Example 1

[0060] 10 g of soybean lecithin (PC-60), 10 g of DHA oil and 5 g of vitamin A are weighed, fully dissolved in 100 mL of anhydrous ethanol, and the anhydrous ethanol is removed by rotary evaporation to form an oily film. Then 75 mL of deionized water is added to dissolve the film to form a vitamin A liposome aqueous solution.

[0061] The particle size and particle size distribution, morphology, encapsulation rate and stability are measured. The average particle size of the liposome is 142 nm, the zeta potential is -36.2 mv, the particle size is uniform, and the loading vitamin A encapsulation rate is 96.4%. The liposome is still stable and has no precipitation and stratification after being placed in a 45℃ stability test box for 1 month, and the loading vitamin A encapsulation rate is measured to be 95.8%.

[0062] Example 2

[0063] 10 g of soybean lecithin (PC-90), 20 g of DHA oil and 20 g of lutein ester are weighed, fully dissolved in 200 mL of anhydrous ethanol, and the anhydrous ethanol is removed by rotary evaporation to form an oily film. Then 30 mL of propylene glycol and 120 mL of deionized water are added to form a water phase to dissolve the film to form a lutein ester liposome aqueous solution.

[0064] The particle size and particle size distribution, morphology, encapsulation rate and stability are measured. The average particle size of the liposome is 169 nm, the zeta potential is -40.3 mv, the particle size is uniform, and the loading lutein ester encapsulation rate is 87.3%. The liposome is still stable and has no precipitation and stratification after being placed in a 45℃ stability test box for 1 month, and the loading lutein ester encapsulation rate is measured to be 84.9%.

[0065] Example 3

[0066] Take 10 g of egg yolk lecithin (PC-50), 12 g of DHA oil, and 3 g of β-carotene, dissolve them thoroughly in 100 mL of ethanol, dissolve them under low temperature stirring, add 75 mL of deionized water, turn on the ultrasonic cell disrupter, adjust the ultrasonic power to about 100 W, set the timer to 20 min, and the effective ultrasonic time is 50%, and remove the ether by rotary evaporation to form a water solution of β-carotene liposomes.

[0067] Determine the particle size and particle size distribution, morphology, encapsulation rate, stability, and other indicators. The average particle size of the liposomes is 160 nm, the ζ potential is -31.8 mv, the particle size is uniform, and the β-carotene loading encapsulation rate is 93.6%. The liposomes are still stable and have no precipitation after being placed in a 45°C stability test box for 1 month, and the measured β-carotene loading encapsulation rate is 92.0%.

[0068] Example 4

[0069] Take 10 g of soy lecithin (PC-80), 16 g of DHA oil, and 10 g of astaxanthin oil, heat and stir under nitrogen protection at 60°C, add 5 g of polyethylene glycol 2000 and 73 mL of deionized water to form an aqueous phase, and continue to stir for 10 min, and then homogenize 5 times at 800 bar with a high-pressure homogenizer to prepare astaxanthin liposomes.

[0070] Determine the particle size and particle size distribution, morphology, encapsulation rate, stability, and other indicators. The average particle size of the liposomes is 104 nm, the ζ potential is -30.7 mv, the particle size is uniform, and the astaxanthin loading encapsulation rate is 99.9%. The liposomes are still stable and have no precipitation after being placed in a 45°C stability test box for 1 month, and the measured astaxanthin loading encapsulation rate is 99.8%.

[0071] Example 5

[0072] Take 10 g of soy lecithin (PC-70), 20 g of DHA oil, and 5 g of coenzyme Q10, heat and stir under nitrogen protection at 60°C, add 63 mL of deionized water and 20 g of glycerol to form an aqueous solution, and continue to stir for 10 min, and then homogenize 3 times at 2200 bar with a micro-jet homogenizer to prepare coenzyme Q10 liposomes.

[0073] Determine the particle size and particle size distribution, morphology, encapsulation rate, stability, and other indicators. The average particle size of the liposomes is 82 nm, the ζ potential is -36.4 mv, the particle size is uniform, and the coenzyme Q10 loading encapsulation rate is 98.4%. The liposomes are still stable and have no precipitation after being placed in a 45°C stability test box for 1 month, and the measured astaxanthin loading encapsulation rate is 97.7%.

[0074] Comparative Example 1

[0075] Take 10 g of soy lecithin (PC-60), 5 g of vitamin A, and dissolve them thoroughly in 100 mL of anhydrous ethanol. Remove the anhydrous ethanol by rotary evaporation to form an oily film. Add 75 mL of deionized water to dissolve the film to form a vitamin A liposome aqueous solution.

[0076] Determine the particle size and particle size distribution, morphology, encapsulation rate, stability, and other indicators. The average particle size of the liposome is 163 nm, the zeta potential is -5.7 mv, the particle size is uniform, and the encapsulation rate of the loaded vitamin A is 85.9%. After being placed in a 45°C stability test box for 1 month, the liposome precipitates.

[0077] Comparative Example 2

[0078] Take 10 g of soy lecithin (PC-60), 10 g of oleic acid, and 5 g of vitamin A, and dissolve them thoroughly in 100 mL of anhydrous ethanol. Remove the anhydrous ethanol by rotary evaporation to form an oily film. Add 75 mL of deionized water to dissolve the film to form a vitamin A liposome aqueous solution.

[0079] Determine the average particle size of the liposome to be 216 nm, the encapsulation rate of the loaded vitamin A to be 90.4%, the zeta potential to be -13.6 mv, and the liposome to precipitate after being placed in a 45°C stability test box for 1 month.

[0080] Comparative Example 3

[0081] Take 10 g of egg yolk lecithin (PC-50), 2 g of DHA oil, and 3 g of β-carotene, and dissolve them thoroughly in 100 mL of ether. Stir and dissolve under low temperature. Add 75 mL of deionized water. Connect the ultrasonic cell disruptor and adjust the ultrasonic power to about 100 W. Set the timer to 20 min and the effective ultrasonic time to 50%. Remove the ether by rotary evaporation to form a β-carotene liposome aqueous solution.

[0082] Determine the particle size and particle size distribution, morphology, encapsulation rate, stability, and other indicators. The average particle size of the liposome is 196 nm, the zeta potential is -19.8 mv, and the encapsulation rate of the loaded β-carotene is 90.3%. After being placed in a 45°C stability test box for 1 month, the average particle size of the liposome increases to 367 nm and the encapsulation rate of the loaded β-carotene is 85.1%.

[0083] Comparative Example 4

[0084] Take 10 g of soy lecithin (PC-80) and 10 g of astaxanthin oil. Heat and stir under nitrogen protection at 60°C. Add 5 g of polyethylene glycol 2000 and 73 mL of deionized water to form an aqueous phase. Stir continuously for 10 min. Homogenize 5 times at 800 bar using a high-pressure homogenizer to prepare astaxanthin liposomes.

[0085] The particle size and particle size distribution, morphology, encapsulation efficiency, stability and other indicators were determined. The average particle size of the liposome was 146 nm, the zeta potential was -10.7 mv, and the astaxanthin loading rate was 97.4%. The liposome was placed in a 45°C stability test box for 1 month and precipitated.

[0086] Comparative Example 5

[0087] Soybean lecithin (PC-60) 5 g, DHA oil 25 g, and vitamin A 5 g were weighed, dissolved in 100 mL of anhydrous ethanol, and rotary evaporated to remove the anhydrous ethanol to form an oily film. Then, 75 mL of deionized water was added to dissolve the film to form a vitamin A liposome aqueous solution.

[0088] The particle size and particle size distribution, morphology, encapsulation efficiency, stability and other indicators were determined. The average particle size of the liposome was 315 nm, the zeta potential was -13.6 mv, the vitamin A loading rate was 86.3%, and the liposome precipitated after being placed in a 45°C stability test box overnight.

[0089] Comparative Example 6

[0090] Soybean lecithin (PC-60) 10 g, phospholipid type DHA (Ji Bi You, prepared according to the method described in 2021106627859 Example 1, DHA content 27%) 10 g, and vitamin A 5 g were weighed, dissolved in 100 mL of anhydrous ethanol, and rotary evaporated to remove the anhydrous ethanol to form an oily film. Then, 75 mL of deionized water was added to dissolve the film to form a vitamin A liposome aqueous solution.

[0091] The particle size and particle size distribution, morphology, encapsulation efficiency, stability and other indicators were determined. The average particle size of the liposome was 135 nm, the zeta potential was -23.6 mv, and the vitamin A loading rate was 95.3%. The average particle size of the liposome increased to 467 nm after being placed in a 45°C stability test box for 1 month, and the vitamin A loading rate was 65.6%.

[0092] The above is only a preferred embodiment of the present application, and it should be noted that those skilled in the art can make several improvements and refinements without departing from the principles of the present application, and these improvements and refinements should also be considered within the scope of protection of the present application.

Claims

1. A liposome comprising a phospholipid bilayer vesicular structure, characterized in that, The composition of the phospholipid bilayer vesicular structure includes phospholipids and long-chain polyunsaturated fatty acid oil; the mass ratio of the phospholipids to the long-chain polyunsaturated fatty acid oil is 1:(1-4); The long-chain polyunsaturated fatty acid oil is DHA oil; the content of DHA in the DHA oil is 35%-70%; The content of triglyceride in the long-chain polyunsaturated fatty acid oil is greater than 95%; The phospholipids are soybean lecithin and / or egg yolk lecithin; The liposome further includes an active ingredient; the active ingredient is one or more of vitamin A, vitamin A ester, vitamin E, vitamin E ester, vitamin D2, vitamin D3, vitamin K, biotin, coenzyme Q10, curcumin, beta-carotene, lutein, canthaxanthin, lutein ester, lycopene and astaxanthin.

2. The liposome of claim 1, wherein, The particle size of the liposome is 80-180 nm.

3. The liposome of claim 1, wherein, The DHA oil further includes one or more of ALA, EPA, LA, AA and DPA.

4. The liposome according to any one of claims 1 to 3, wherein, The liposome further includes a co-surfactant and / or a surfactant; The co-surfactant is a polyol and / or a polyethylene glycol; The polyol is one or more of propylene glycol, butylene glycol and glycerol; The polyethylene glycol is one or more of polyethylene glycol 400, polyethylene glycol 1000 and polyethylene glycol 2000; The surfactant is one or more of Tween 20, Tween 40, Tween 60, Tween 80, PEG-10 hydrogenated castor oil, PEG-20 hydrogenated castor oil, PEG-30 hydrogenated castor oil, PEG-40 hydrogenated castor oil, PEG-50 hydrogenated castor oil and PEG-60 hydrogenated castor oil.

5. The liposome of claim 4, wherein, The mass ratio of the phospholipids, the long-chain polyunsaturated fatty acid oil and the active ingredient is 1:(1-4):(0.1-2).

6. A method of preparing the liposome according to any one of claims 1 to 5, characterized in that, The method includes the following steps: S1, weighing the liposome composition ingredients to obtain the mass ratio; the composition of the liposome includes the phospholipids, the long-chain polyunsaturated fatty acid oil and the active ingredient; S2, mixing the liposome composition ingredients in step S1 with anhydrous ethanol to obtain a mixed solution; S3, evaporating the alcohol in the mixed solution in step S2 to obtain an oily film; S4, placing the oily film in step S3 in a dispersion medium or a dispersion medium containing the surfactant to obtain a liposome dispersion system; the dispersion medium includes water and / or the co-surfactant.

7. A liposome, characterized by, The liposome includes the liposome dispersion system in step S4 of claim 6 and a fragrance; the mass fraction of the fragrance in the liposome is 0.001%-0.1%.