Composite embedded vitamin liposome as well as preparation method and application thereof

Abstract

The invention discloses a composite embedded vitamin liposome as well as a preparation method and application thereof, by optimizing a formula and a process, adopting high-purity natural plant source phospholipid, a proper amount of a wall material and a membrane stabilizer and combining an antioxidant, the vitamin retention rate exceeds 95% after the liposome is accelerated at 60 DEG C for 20 days, and the rehydration particle size is between 200 and 800nm; the flowability is good, no bad taste or smell exists, and efficient embedding and stable release of vitamins are realized. The liposome has excellent physical and chemical stability and is suitable for various dosage forms such as tablets and hard-shell capsules, and the bioavailability and market application value of the product are remarkably improved. In addition, a clean solvent ethanol is used in the preparation process and is effectively removed and recycled, so that the method meets the requirements of food processing industry laws and regulations, is environment-friendly and is suitable for large-scale production.

Description

Technical Field

[0001] The present invention belongs to the field of food nutrition and health, and particularly relates to a composite-embedded vitamin liposome and a preparation method thereof. Through unique formulation design and optimized preparation processes, this composite achieves efficient embedding and stable release of vitamins, significantly enhancing the bioavailability and market application value of the product. Background Art

[0002] As essential nutrients for the human body, vitamins play an important role in the food industry. However, traditional vitamin preparations have problems such as poor stability and low bioavailability, especially in the mixed application of water-soluble and fat-soluble vitamins. As a nano-drug delivery system, liposomes, due to their unique bilayer membrane structure, can effectively improve the stability and bioavailability of drugs and have been widely used in the pharmaceutical field. In recent years, liposome technology has also been gradually introduced into the food industry to improve the delivery efficiency of functional ingredients.

[0003] As an advanced drug delivery system, liposome technology has quickly become a research hotspot in multiple directions since it was first reported in the 1960s. Liposomes are vesicle structures formed by phospholipid bilayer membranes encapsulating hydrophilic or hydrophobic components, and their diameters are usually between dozens of nanometers and several micrometers. This unique structure endows liposomes with many superior properties, such as good biocompatibility and adjustable particle size.

[0004] Liposome technology has been widely used to improve the stability of drug components, increase the targeting of drugs, extend the action time of drugs, and reduce the side effects of drugs. By encapsulating drug molecules inside liposomes, the drugs can be effectively protected from the external environment, such as enzymatic hydrolysis and oxidation, thereby improving the stability and bioavailability of the drugs. In addition, the phospholipid molecules on the surface of liposomes are similar to the cell membrane structure, enabling liposomes to more easily penetrate cell membranes and improve the transdermal absorption and cell uptake efficiency of drugs.

[0005] With the acceleration of the modern life rhythm and the increase in work pressure, the sub-healthy state is widespread, and consumers' demand for health management is increasing day by day. At the same time, the state has strengthened the supervision of the dietary supplement industry, and the policies and regulations have been gradually improved, providing strong guarantees for the healthy development of the industry. Liposome preparations are also increasingly used in foods and health products to solve problems such as poor stability, poor absorption, low bioavailability of nutrients, and easy gastrointestinal discomfort.

[0006] Patent CN115006410A discloses a compound vitamin liposome, its preparation method and application. During the preparation process, components such as vitamins need to be heated and melted at 150 - 160°C. This condition will cause a large amount of degradation of vitamins, and the degradation products may have adverse effects on the human body, making it difficult to ensure product quality. Patent CN115737635A discloses a VC liposome complex and its preparation method. The phospholipids selected are one or a mixture of phospholipids such as lecithin, cephalin, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, etc. Different types of phospholipids have large differences in solubility and critical packing parameter (CPP), which will lead to instability between product batches and is difficult to apply to large-scale production. Patent CN109497561A discloses a preparation method of vitamin B 12 nano-liposome. The traditional thin film hydration method is used, but this method is limited to laboratory preparation and there is no corresponding production-level equipment, lacking the ability to meet industrialization requirements.

[0007] Most existing liposome vitamin complexes adopt single preparation methods, and these methods have certain limitations in particle size control, stability, and production efficiency. Summary of the Invention

[0008] In order to overcome the above deficiencies, the present invention aims to provide a compound-embedded vitamin liposome, its preparation method and application. Through an optimized formula and process combination form, by innovatively combining the ethanol injection process and the spray drying process, it aims to provide a liposome vitamin complex with good fluidity, small particle size, high encapsulation rate, good instant solubility, and significantly improved bioavailability. This process improves the stability of liposomes, simplifies the production process, reduces production costs, and makes it more suitable for industrial production of food and pharmaceutical preparations. The compound-embedded liposome prepared by the present invention has good fluidity and small particle size, and can be applied to directions such as tableting, hard shell capsules, and solid beverages.

[0009] The first aspect of the present invention is to protect a compound-embedded vitamin liposome, which is prepared from the following raw materials by weight: 0.1 - 80 parts of vitamins, 10 - 60 parts of phospholipids, 0.1 - 3 parts of antioxidants, 1 - 12 parts of membrane layer stabilizers, and 3 - 25 parts of wall materials. Further preferably: 0.1 - 75 parts of vitamins, 15 - 50 parts of phospholipids, 0.1 - 2 parts of antioxidants, 2 - 10 parts of membrane layer stabilizers, and 5 - 20 parts of wall materials. The most preferred value range: vitamin content: 20 - 75 parts, phospholipid content: 20 - 40 parts, antioxidant content: 0.5 - 1 part, membrane layer stabilizer content: 2 - 6 parts, wall material content: 10 - 15 parts;

[0010] For the above technical solution, further preferably: the vitamins include water-soluble vitamins and / or fat-soluble vitamins.

[0011] For the above technical solution, more preferably: the water-soluble vitamins: 10-75 parts; fat-soluble vitamins: 0.5-20 parts; more preferably: water-soluble vitamins 45-75 parts, fat-soluble vitamins 2-15 parts.

[0012] For the above technical solution, it can also meet the requirement of compound use of vitamins. More preferably: the ratio of the water-soluble vitamins to the fat-soluble vitamins is 3-15:1. This ratio range can ensure good embedding rate and stability, and at the same time meet the requirements of different application scenarios. Further preferably, the ratio of water-soluble vitamins to fat-soluble vitamins is 4-10:1.

[0013] For the above technical solution, more preferably: the water-soluble vitamins can be selected from one or more of vitamin C, vitamin B1, vitamin B2, vitamin B6, and vitamin B12.

[0014] For the above technical solution, more preferably: the fat-soluble vitamins can be selected from one or more of vitamin A, vitamin D, vitamin E, and vitamin K.

[0015] For the above technical solution, more preferably: the phospholipids are selected from one or more of soybean lecithin, sunflower lecithin, and rapeseed lecithin.

[0016] For the above technical solution, more preferably: the content of acetone-insoluble matter in the phospholipids is required to be greater than 60%. Further preferably, the content of acetone-insoluble matter is greater than 90%. More preferably, the content of acetone-insoluble matter is greater than 95%.

[0017] For the above technical solution, more preferably: the content of phosphatidylcholine in the above-mentioned phospholipids is required to be greater than 50%. Further preferably, the content of phosphatidylcholine is greater than 70%. More preferably, the content of phosphatidylcholine is greater than 90%. In the examples, high-purity phospholipids such as soybean lecithin PC90 and above are selected.

[0018] For the above technical solution, more preferably: the film layer stabilizer includes one or more of phytosterol, mono- and diglycerol fatty acid esters, fatty acids, and vegetable oils.

[0019] For the above technical solution, more preferably: the weight ratio of the film layer stabilizer to the phosphatidylcholine component in the phospholipids is 1:3-1:15. Further preferably, the weight ratio of the film layer stabilizer to the phosphatidylcholine component in the phospholipids is 1:3-1:12. More preferably, it is 1:4-1:10; the most preferred weight ratio is 1:4-1:8.

[0020] For the above technical solutions, more preferably: The phytosterol is selected from one or more of sitosterol, stigmasterol, brassicasterol, ergosterol, and microalgae sterol.

[0021] For the above technical solutions, more preferably: To meet the stability of liposomes, it is required that the melting points of the above-mentioned mono- and diglycerol fatty acid esters, fatty acids, and vegetable oils are in the range of 40-90 °C, preferably 40-80 °C, and more preferably 45-65 °C.

[0022] For the above technical solutions, more preferably: The antioxidant is selected from one or more of ascorbic acid, ascorbyl palmitate, mixed tocopherols, citrus flavonoids, proanthocyanidins, tea polyphenols, and carotenoids.

[0023] For the above technical solutions, more preferably: The wall material includes one or more of modified starch, gum arabic, maltodextrin, resistant dextrin, maltooligosaccharide, and isomaltooligosaccharide.

[0024] Another aspect of the present invention is to protect a method for preparing a composite-embedded vitamin liposome, which includes the following steps:

[0025] S1. Preparation of the aqueous phase: Prepare an aqueous solution of water-soluble vitamins;

[0026] S2. Preparation of the alcohol phase: After anhydrous ethanol, phospholipids, and membrane layer stabilizers are fully mixed and dissolved, fat-soluble vitamins are added until completely dissolved;

[0027] S3. Preparation of the wall material solution: Prepare an aqueous solution of the wall material;

[0028] S4. Slowly inject the alcohol phase into the aqueous phase, stir well to form a primary liposome dispersion solution. After concentration under reduced pressure, the concentrated solution is slowly added to the aqueous solution of the wall material obtained in S3, and stirred well to form a composite-embedded liposome dispersion solution.

[0029] S5. Perform high-speed shearing, high-pressure homogenization, and drying on the composite-embedded liposome dispersion solution obtained in S4 to obtain the product.

[0030] For the technical solutions described above, preferably, the dissolution temperature of the water-soluble vitamins in S1 is 30-70 °C; more preferably 40-60 °C; most preferably 45-55 °C.

[0031] Further preferably, to reduce the degradation of the content of water-soluble vitamins during the sample preparation process, the dissolution process of the water-soluble vitamins in S1 needs to be carried out in a light-shielded and inert gas environment.

[0032] For the technical solutions described above, preferably, the dissolution temperature of the phospholipid and the membrane layer stabilizer in S2 is 40-70 °C; more preferably 50-65 °C; most preferably 55-60 °C.

[0033] Further preferably, in order to reduce the degradation of the content of fat-soluble vitamins during the sample preparation process, in step S2, before adding the fat-soluble vitamins, the temperature of the alcohol phase is adjusted to 35-60 °C and maintained until the fat-soluble vitamins are completely dissolved. More preferably 40-50 °C. Further preferably, the dissolution temperature of the fat-soluble vitamins in the alcohol phase is 40-50 °C, and the dissolution process needs to be carried out in a light-shielded and inert gas environment.

[0034] For the technical solutions described above, the weight ratio of the amount of deionized water used in S1 to the amount of absolute ethanol used in S2 should be within a certain range. Preferably, the weight ratio of water to ethanol is (1-8):1. Further preferably, the weight ratio of water to ethanol is (1-5):1. More preferably, it is (2-4):1.

[0035] For the technical solutions described above, preferably, the solid content of the wall material solution in S3 is 20-60 wt%; more preferably 30-50 wt%; most preferably 35-45 wt%.

[0036] For the technical solutions described above, preferably, the temperature of the vacuum concentration process in S4 is 30-70 °C; more preferably 40-60 °C. Further preferably, the temperature is 45-55 °C.

[0037] For the technical solutions described above, preferably, the vacuum degree of the vacuum concentration process in S4 is -0.07 to -0.1 MPa; preferably -0.1 to -0.08 MPa. Further preferably, the vacuum degree is -0.09 to -0.085 MPa.

[0038] For the technical solutions described above, preferably, the stirring temperature after adding to the aqueous wall material solution in S4 is 40-60 °C. Further preferably, the temperature is 40-50 °C.

[0039] For the technical solutions described above, preferably, the homogenization pressure in S5 is 20-40 MPa; further preferably, the homogenization pressure is 30-40 MPa; preferably, the number of homogenization times is two.

[0040] For the technical solutions described above, the drying method in S5 can be selected from one of spray drying, low-temperature spray drying, freeze drying, and freeze spray drying. Preferably, in order to better meet the needs of industrialization, the drying method in S5 is selected as spray drying or low-temperature spray drying. Further preferably, for vitamin components with poor thermal stability, the drying method is selected as low-temperature spray drying, and for vitamin components with good thermal stability, the drying method is selected as spray drying.

[0041] Finally, the present invention also protects the application of the above-mentioned composite-embedded vitamin liposomes, and the application fields include the production of tablet preparations, hard-shell capsules, solid beverages, etc. Among them, solid beverages include fruit and vegetable solid beverages, protein solid beverages, tea solid beverages, plant solid beverages, etc.

[0042] Advantages of the present invention:

[0043] 1. The composite-embedded vitamin liposomes prepared by the present invention have good fluidity, uniform particle size, no bad smell, and the rehydrated particle size is in the range of 200-800 nm.

[0044] 2. The composite-embedded vitamin liposomes prepared by the present invention have very good physical stability and chemical stability after secondary embedding. After the obtained liposomes are accelerated at 60°C for 20 days, problems such as caking, browning, and deterioration of smell do not occur, and the content degradation rate is less than 5%.

[0045] 3. For the composite-embedded vitamin liposomes prepared by the present invention, the phospholipids in the raw materials are all natural plant-derived components, and cholesterol is not added to the formula, which better meets the needs of consumers.

[0046] 4. The preparation method of the composite-embedded vitamin liposomes involved in the present invention uses clean solvent ethanol as a processing aid and removes and recovers it in the process. The solvent residue of the product is less than 50 ppm, which meets the requirements of food processing industry regulations and does not cause environmental pollution.

[0047] 5. The preparation method of the composite-embedded vitamin liposomes involved in the present invention simplifies the complex steps of traditional liposome preparation, reduces the production difficulty and cost, and improves the production efficiency; it has very good industrial adaptability and can meet the large-scale production of products.

[0048] 6. The composite-embedded vitamin liposomes prepared by the present invention have a wide range of applications and can be widely applied to various dietary supplements and functional health food fields such as tablet preparations, hard-shell capsules, and solid beverages.

[0049] 7. The composite-embedded vitamin liposomes prepared by the present invention can significantly improve the bioavailability of vitamins and ensure that the human body can more efficiently absorb and utilize vitamin components.

[0050] 8. The composite-embedded vitamin liposomes prepared by the present invention can be rapidly dispersed and dissolved in water or other media to form a uniform and stable solution, which is suitable for the application requirements of various dosage forms. Detailed Embodiments

[0051] The present invention will be further described below in conjunction with embodiments, but it should be understood that the protection scope of the present invention is not limited by the embodiments.

[0052] Example 1

[0053] Aqueous phase: Weigh 225 parts of deionized water, heat it to 50 °C, add 75 parts of vitamin C and 0.5 part of citrus flavonoids, and stir until completely dissolved.

[0054] Alcohol phase: Weigh 40 parts of absolute ethanol, add 15 parts of soybean lecithin PC90 (PC90: phosphatidylcholine content ≥ 90%, the same below) and 2 parts of mono- and diglycerol fatty acid esters, heat it to 60 °C, and stir until completely dissolved.

[0055] Wall material solution: Add 7.5 parts of modified starch to 14 parts of deionized water, heat and stir until completely dissolved.

[0056] Slowly inject the alcohol phase into the aqueous phase, stir well, and then transfer it to a rotary evaporation flask for concentration under reduced pressure. The concentration conditions are -0.09 MPa and 55 °C.

[0057] Add the vitamin C liposome concentrate to the wall material solution, stir at 45 °C for 1 hour. Shear the feed liquid at a high speed of 5500 revolutions per minute for 2 minutes, and homogenize it at 35 MPa for 2 times. Then carry out spray drying, control the inlet air temperature at 130 °C and the outlet air temperature at 80 °C to obtain vitamin C liposome microcapsule powder.

[0058] Example 2

[0059] Aqueous phase: Weigh 200 parts of deionized water, heat it to 50 °C, add 45 parts of vitamin B6 and 1 part of grape seed extract, and stir until completely dissolved.

[0060] Alcohol phase: Weigh 90 parts of absolute ethanol, add 35 parts of sunflower lecithin PC70 and 3 parts of palm oil, heat it to 60 °C, and stir until completely dissolved.

[0061] Wall material solution: Add 16 parts of arabic gum to 37 parts of deionized water, heat and stir until completely dissolved.

[0062] Slowly inject the alcohol phase into the aqueous phase, stir well, and then transfer it to a rotary evaporation flask for concentration under reduced pressure. The concentration conditions are -0.1 MPa and 50 °C.

[0063] Add the vitamin B6 liposome concentrate to the wall material solution and stir at 40 °C for 1 hour. Shear the feed liquid at a high speed of 5000 revolutions per minute for 3 minutes and homogenize it under high pressure of 40 MPa twice. Then carry out spray drying, control the inlet air temperature at 140 °C and the outlet air temperature at 82 °C to obtain vitamin B6 liposome microcapsule powder.

[0064] Example 3

[0065] Aqueous phase: Weigh 200 parts of deionized water, heat it up to 50 °C, add 1 part of tea polyphenols, and stir until completely dissolved.

[0066] Alcohol phase: Weigh 70 parts of absolute ethanol, add 2 parts of vitamin D3 crystals, 25 parts of soybean lecithin PC70 and 3 parts of stigmasterol, heat it up to 60 °C, and stir until completely dissolved.

[0067] Wall material solution: Add 8 parts of modified starch and 62 parts of maltodextrin to 140 parts of deionized water, heat and stir until completely dissolved.

[0068] Slowly inject the alcohol phase into the aqueous phase, stir well to mix evenly, and then transfer it to a rotary evaporation flask for concentration under reduced pressure. The concentration conditions are -0.09 MPa and 50 °C.

[0069] Add the vitamin D3 liposome concentrate to the wall material solution and stir at 40 °C for 1 hour. Shear the feed liquid at a high speed of 5000 revolutions per minute for 3 minutes and homogenize it under high pressure of 40 MPa twice. Then carry out spray drying, control the inlet air temperature at 140 °C and the outlet air temperature at 82 °C to obtain vitamin D3 liposome microcapsule powder.

[0070] Example 4

[0071] Aqueous phase: Weigh 200 parts of deionized water, heat it up to 50 °C, and set aside.

[0072] Alcohol phase: Weigh 120 parts of absolute ethanol, 55 parts of rapeseed lecithin PC50 and 4 parts of sitosterol, heat it up to 60 °C, and stir until completely dissolved. Add 8 parts of vitamin E and 0.2 part of ascorbyl palmitate, and stir until completely dissolved.

[0073] Wall material solution: Add 32.8 parts of maltodextrin to 70 parts of deionized water, heat and stir until completely dissolved.

[0074] Slowly inject the alcohol phase into the aqueous phase, stir well to mix evenly, and then transfer it to a rotary evaporation flask for concentration under reduced pressure. The concentration conditions are -0.085 MPa and 55 °C.

[0075] Add the vitamin E liposome concentrate to the wall material solution and stir at 40 °C for 1 hour. High-speed shear the feed liquid at 5000 revolutions per minute for 3 minutes and perform high-pressure homogenization at 40 MPa twice. Subsequently, perform spray drying, controlling the inlet air temperature at 140 °C and the outlet air temperature at 82 °C to obtain vitamin E liposome microcapsule powder.

[0076] Example 5

[0077] Aqueous phase: Weigh 180 parts of deionized water, heat it to 50 °C, add 20 parts of vitamin B6, 10 parts of vitamin B12, and 2 parts of ascorbic acid, and stir until completely dissolved.

[0078] Alcohol phase: Weigh 90 parts of absolute ethanol, add 40 parts of sunflower lecithin PC80 and 4 parts of stigmasterol, heat it to 60 °C, and stir until completely dissolved.

[0079] Wall material solution: Add 24 parts of maltodextrin to 50 parts of deionized water, heat and stir until completely dissolved.

[0080] Slowly inject the alcohol phase into the aqueous phase, stir well to mix evenly, and then transfer it to a rotary evaporation flask for concentration under reduced pressure. The concentration conditions are -0.09 MPa and 48 °C.

[0081] Add the compound vitamin B liposome concentrate to the wall material solution and stir at 40 °C for 1 hour. High-speed shear the feed liquid at 5000 revolutions per minute for 3 minutes and perform high-pressure homogenization at 40 MPa twice. Subsequently, perform spray drying, controlling the inlet air temperature at 140 °C and the outlet air temperature at 82 °C to obtain compound vitamin B liposome microcapsule powder.

[0082] Example 6

[0083] Aqueous phase: Weigh 260 parts of deionized water, heat it to 50 °C, and set aside.

[0084] Alcohol phase: Weigh 150 parts of absolute ethanol, add 50 parts of soy lecithin PC95 and 5 parts of mono- and diglycerol fatty acid esters, heat it to 60 °C, and stir until completely dissolved. Add 15 parts of vitamin A, 5 parts of vitamin D crystals, and 0.5 part of mixed tocopherols, and stir until completely dissolved.

[0085] Wall material solution: Add 10 parts of resistant dextrin and 14.5 parts of isomaltooligosaccharide to 50 parts of deionized water, heat and stir until completely dissolved.

[0086] Slowly inject the alcohol phase into the aqueous phase, stir well to mix evenly, and then transfer it to a rotary evaporation flask for concentration under reduced pressure. The concentration conditions are -0.09 MPa and 48 °C.

[0087] Add the compound vitamin A and D liposome concentrate to the wall material solution and stir at 40 °C for 1 hour. Shear the feed liquid at a high speed of 5000 revolutions per minute for 3 minutes and homogenize it at a high pressure of 40 MPa for 2 times. Then carry out spray drying, control the inlet air temperature at 140 °C and the outlet air temperature at 82 °C to obtain the compound vitamin A and D liposome microcapsule powder.

[0088] Example 7

[0089] Aqueous phase: Weigh 150 parts of deionized water, heat it to 50 °C, add 45 parts of vitamin C, and stir until it is completely dissolved.

[0090] Alcohol phase: Weigh 80 parts of absolute ethanol, add 30 parts of rapeseed lecithin PC70 and 2 parts of stearic acid, heat it to 60 °C, and stir until it is completely dissolved. Add 10 parts of vitamin E and 0.2 part of ascorbyl palmitate, and stir until it is completely dissolved.

[0091] Wall material solution: Add 5 parts of gum arabic and 7.8 parts of maltooligosaccharide to 30 parts of deionized water, heat and stir until it is completely dissolved.

[0092] Slowly inject the alcohol phase into the aqueous phase, stir well and mix evenly, and then transfer it to a rotary evaporation flask for concentration under reduced pressure. The concentration conditions are -0.09 MPa and 52 °C.

[0093] Add the compound vitamin liposome concentrate to the wall material solution and stir at 40 °C for 1 hour. Shear the feed liquid at a high speed of 5000 revolutions per minute for 3 minutes and homogenize it at a high pressure of 40 MPa for 2 times. Then carry out spray drying, control the inlet air temperature at 140 °C and the outlet air temperature at 82 °C to obtain the compound vitamin liposome microcapsule powder.

[0094] In the above Examples 1 to 7, the following parameters were screened, as shown in Table 1:

[0095] Table 1

[0096]

[0097] Comparative Example 1 (adding low-specification phospholipids)

[0098] On the basis of Example 5, the influence of the phospholipid specification on the product quality was compared. In this comparative example, low-specification phospholipids were used.

[0099] Aqueous phase: Weigh 180 parts of deionized water, heat it to 50 °C, add 20 parts of vitamin B6, 10 parts of vitamin B12 and 2 parts of ascorbic acid, and stir until it is completely dissolved.

[0100] Alcohol phase: Weigh 90 parts of absolute ethanol, add 40 parts of powdered sunflower phospholipids (phosphatidylcholine content ≈ 20%, and other phospholipid components such as phosphatidylinositol and phosphatidylethanolamine), and 4 parts of stigmasterol. Heat to 60 °C and stir. Since there are a large number of ethanol-insoluble substances in the powdered phospholipids, add chloroform and keep stirring until completely dissolved.

[0101] Wall material solution: Add 24 parts of maltodextrin to 50 parts of deionized water, heat and stir to completely dissolve it.

[0102] Slowly inject the alcohol phase into the aqueous phase, stir well to mix evenly, and then transfer it to a rotary evaporation flask for concentration under reduced pressure. The concentration conditions are -0.09 MPa and 48 °C.

[0103] Add the compound vitamin B liposome concentrate to the wall material solution, and stir at 40 °C for 1 hour. Shear the feed liquid at a high speed of 5000 revolutions per minute for 3 minutes, and homogenize it at 40 MPa for 2 times. Then carry out spray drying, control the inlet air temperature at 140 °C and the outlet air temperature at 82 °C to obtain the compound vitamin B liposome microcapsule powder.

[0104] Comparative example 2 (without adding wall material)

[0105] On the basis of Example 2, compare the influence of the wall material on the product properties. In this comparative example, no wall material component is added.

[0106] Aqueous phase: Weigh 200 parts of deionized water, heat to 50 °C, add 45 parts of vitamin B6 and 1 part of grape seed extract, and stir to completely dissolve it.

[0107] Alcohol phase: Weigh 90 parts of absolute ethanol, add 35 parts of sunflower lecithin PC70 and 3 parts of palm oil, heat to 60 °C, and stir until completely dissolved.

[0108] Slowly inject the alcohol phase into the aqueous phase, stir well to mix evenly, and then transfer it to a rotary evaporation flask for concentration under reduced pressure. The concentration conditions are -0.1 MPa and 50 °C. Adjust the solid content to 35%, and stir at 40 °C for 1 hour. Shear the feed liquid at a high speed of 5000 revolutions per minute for 3 minutes, and homogenize it at 40 MPa for 2 times. Then carry out spray drying, control the inlet air temperature at 140 °C and the outlet air temperature at 82 °C to obtain the vitamin B6 liposome microcapsule powder.

[0109] Comparative example 3 (without adding film layer stabilizer)

[0110] On the basis of Example 3, compare the influence of the film layer stabilizer on the product quality. In this comparative example, no film layer stabilizer is added.

[0111] Aqueous phase: Weigh 200 parts of deionized water, heat to 50 °C, add 1 part of tea polyphenols, and stir to completely dissolve it.

[0112] Alcohol phase: Weigh 70 parts of absolute ethanol, add 2 parts of vitamin D3 crystals and 25 parts of soy lecithin PC70, heat to 60 °C, and stir until completely dissolved.

[0113] Wall material solution: Add 8 parts of modified starch and 62 parts of maltodextrin to 140 parts of deionized water, heat and stir until completely dissolved.

[0114] Slowly inject the alcohol phase into the water phase, stir well, and then transfer it to a rotary evaporation flask for concentration under reduced pressure. The concentration conditions are -0.09 MPa and 50 °C.

[0115] Add the vitamin D3 liposome concentrate to the wall material solution, stir at 40 °C for 1 hour. Shear the feed liquid at a high speed of 5000 revolutions per minute for 3 minutes, and homogenize it at 40 MPa for 2 times. Then carry out spray drying, control the inlet air temperature at 140 °C and the outlet air temperature at 82 °C to obtain vitamin D3 liposome microcapsule powder.

[0116] Comparative Example 4 (without adding antioxidant)

[0117] On the basis of Example 6, compare the influence of antioxidants on the product stability. In this comparative example, no antioxidant is added.

[0118] Water phase: Weigh 260 parts of deionized water, heat to 50 °C, and set aside.

[0119] Alcohol phase: Weigh 150 parts of absolute ethanol, add 50 parts of soy lecithin PC95 and 5 parts of mono- and diglycerol fatty acid esters, heat to 60 °C, and stir until completely dissolved. Add 15 parts of vitamin A and 5 parts of vitamin D crystals, and stir until completely dissolved.

[0120] Wall material solution: Add 10 parts of resistant dextrin and 14.5 parts of isomaltooligosaccharide to 50 parts of deionized water, heat and stir until completely dissolved.

[0121] Slowly inject the alcohol phase into the water phase, stir well, and then transfer it to a rotary evaporation flask for concentration under reduced pressure. The concentration conditions are -0.09 MPa and 48 °C.

[0122] Add the compound vitamin AD liposome concentrate to the wall material solution, stir at 40 °C for 1 hour. Shear the feed liquid at a high speed of 5000 revolutions per minute for 3 minutes, and homogenize it at 40 MPa for 2 times. Then carry out spray drying, control the inlet air temperature at 140 °C and the outlet air temperature at 82 °C to obtain compound vitamin A and D liposome microcapsule powder.

[0123] Application Example 1 (preparing tablet preparation)

[0124] Take 60 parts of the liposome samples of the above-mentioned examples or comparative examples, 23.5 parts of sorbitol, 15 parts of microcrystalline cellulose, and 1.5 parts of magnesium stearate, mix them evenly, and then perform direct compression tableting to test the tablet indexes.

[0125] Application Example 2 (preparation of hard capsules)

[0126] Take the liposome samples of the above-mentioned examples or comparative examples and directly pour them into the capsule shells to prepare hard capsules, and test the tablet indexes.

[0127] Application Example 3 (preparation of solid beverages)

[0128] Take 35 parts of the liposome samples of the above-mentioned examples or comparative examples, 30 parts of erythritol, 25 parts of maltodextrin, 9.4 parts of stevioside, 0.5 parts of citric acid, and 0.1 part of edible essence, mix them evenly, and then perform granulation. The net content is 3 g / package to prepare solid beverages.

[0129] Effect Comparison Group 1. Index Evaluation

[0130] Perform index evaluation on the liposome samples prepared in Examples 1-7 and Comparative Examples 1-4. The evaluation indexes include product state, color, taste and smell, embedding rate, particle size, angle of repose, solvent residue, and rehydrated particle size.

[0131] State: Visual inspection.

[0132] Color: Visual inspection.

[0133] Taste and smell: Sniffing.

[0134] Particle size: Weigh about 10 g of the sample, place it in a 40-mesh standard sieve, shake for more than 3 min, and calculate the passing rate through the 40-mesh sieve.

[0135] Embedding rate: Determine the content of free vitamins using the ultrafiltration method or the organic solvent extraction method. Embedding rate = (total vitamin content - free vitamin content) / total vitamin content * 100%

[0136] Angle of repose: Measure using an angle of repose measuring instrument.

[0137] Solvent residue: Determine using a headspace sampler combined with gas chromatography.

[0138] Rehydrated particle size: Dissolve the liposomes at a concentration of 1%, and use a nanoparticle size analyzer to measure the particle size after rehydration, and take the average particle size.

[0139] Table 2. Results of Liposome Index Evaluation

[0140]

[0141] According to the evaluation results of liposome indicators, the test results of product status, color, taste and smell, particle size, embedding rate, angle of repose, etc. of different examples and comparative examples can be compared.

[0142] Status: The liposomes of Examples 1-7 are all free-flowing powders, indicating that the fluidity state of the powder is very good. The product state of Comparative Example 2 is sticky and caked. This is because no wall material was added to the formula. During the spray drying process, the drying temperature was much higher than the phase transition temperature of phospholipids, resulting in the phenomenon of sticking and caking of the product, and the yield decreased significantly, which is not convenient for industrial production.

[0143] Color: The color of the liposome samples is affected by the type of vitamin and the formula. The liposomes in Examples 1-7 are off-white to light yellow. In Comparative Example 1, low-purity phospholipids were used, resulting in a yellow color. In Comparative Example 2, no wall material was added, resulting in a yellow color. In fact, the addition of high-purity lecithin and wall material will make the color of the liposome lighter.

[0144] Taste and smell: The liposomes in Examples 1-7 have no obvious smell, while Comparative Examples 1 and 2 have a slight phospholipid smell, which is also due to the use of low-purity lecithin in the formula and the absence of wall material. The higher the purity of lecithin, the lighter the smell, making the liposome more soluble in various terminal dosage forms and more acceptable to consumers. Similarly, the addition of wall material enables the liposome to be double-embedded, which can further mask the smell of phospholipids and delay the oxidation of phospholipids and the degradation of vitamins.

[0145] Embedding rate: The embedding rates of the liposomes in Examples 1-7 are all above 70%. The size of the embedding rate is greatly affected by the product formula. For example, reducing the vitamin loading, increasing the phospholipid specification, using a membrane stabilizer, adding a wall material, etc. will all increase the embedding rate of the product. By comparing the embedding rates of the products in each comparative example and the corresponding examples, the differences in embedding rates caused by the formula composition can be significantly found.

[0146] Particle size: The sieve passing rate of the liposomes in Examples 1-7 through 40 meshes is >99%, and the sieve passing rate of Comparative Example 2 is 90%, indicating that the sticking situation of the product is relatively serious, which is consistent with the analysis results of the state index.

[0147] Angle of repose: The angle of repose of the powder is an important parameter describing the stacking characteristics of the powder in a static state, which reflects the fluidity of the powder. The smaller the angle of repose, the better the fluidity of the powder. The angle of repose of the liposomes in Examples 1-7 is about 40°, indicating very good fluidity, while the angle of repose of Comparative Examples 1 and 2 increases, indicating relatively poor fluidity.

[0148] Solvent residue: This process can effectively remove the organic solvents used in the process, and the solvent residues of the products in each example and comparative example are all less than 50 ppm.

[0149] Redispersion particle size: After the liposomes in Examples 1-7 were redispersed and dissolved, the average particle size was less than 400 nm. Compared with the corresponding examples, the particle sizes of the liposomes in Comparative Examples 1-4 increased to a certain extent, which also indicates the influence of the formulation.

[0150] In addition, the powder sunflower phospholipids (PC20) were used in Comparative Example 1. Since there were a large number of ethanol-insoluble substances in the powder phospholipids, chloroform was added during the dissolution process, which was not a processing aid permitted in GB2760. Therefore, it does not have the conditions for industrialization in the food industry.

[0151] Through the evaluation results of the above partial indicators, it illustrates the necessity of the selection of phospholipid specifications and the addition of wall materials.

[0152] The analysis of the above indicators only reflects the initial product level. Therefore, it is necessary to further compare the accelerated stability of each group.

[0153] Effect comparison group 2. Investigation of accelerated stability

[0154] The liposome samples prepared in Examples 1-7 and Comparative Examples 1-4 were subjected to accelerated investigation at 60 °C, and the product state was observed and the vitamin retention rate was measured.

[0155] Table 3. Evaluation results of indicators after 20 days of liposome acceleration

[0156]

[0157] Sensory indicators: After 20 days of accelerated investigation at 60 °C, the browning of the liposomes in Comparative Examples 1, 2, and 4 was significantly aggravated, and an obvious phospholipid oxidation odor occurred. The reasons are as follows: Low-purity lecithin was used in Comparative Example 1, and phospholipids and glycolipids with more other components would aggravate oxidation; in Comparative Example 2, no wall material was used for secondary embedding, and the phospholipids were directly exposed to the outermost layer, aggravating the process of oxidation and browning; in Comparative Example 4, no antioxidant was added, which aggravated the oxidation process, resulting in product browning and an unpleasant odor.

[0158] Entrapment efficiency: The entrapment efficiencies of the products in each group decreased to a certain extent after 20 days of acceleration. The decrease in the entrapment efficiency of the liposomes in Examples 1-7 was within 10%, while the decrease in the entrapment efficiency of the samples in Comparative Examples 1-4 was very obvious. The reasons are as follows: Low-purity lecithin was used in Comparative Example 1, and leakage of the liposome structure was more likely to occur during the acceleration process; in Comparative Example 2, no wall material was used for secondary embedding, and accelerated phospholipid oxidation led to a decrease in the encapsulation effect; in Comparative Example 3, the lack of a membrane layer stabilizer accelerated the leakage of the content; in Comparative Example 4, phospholipid oxidation also led to a decrease in the entrapment efficiency.

[0159] Vitamin retention rate: After 20 days of accelerated testing at 60°C, the vitamin retention rates of the liposomes in Examples 1 to 7 were all > 95%. However, compared with the corresponding examples, the vitamin retention rates in Comparative Examples 1 to 4 decreased. Especially in Comparative Example 4, due to the absence of antioxidant addition, the VA retention rate was only 92.0%, showing a significant decrease.

[0160] The above results of accelerated stability further illustrate the necessity of choosing phospholipid specifications, adding wall materials, adding stabilizers, and adding antioxidants.

[0161] Effect comparison group 3. Investigation of application performance

[0162] For the liposome samples prepared in Examples 2, 3, 5, 6 and Comparative Examples 1 to 4, tablet preparations, hard-shell capsules, and solid beverages were respectively prepared, and their application performance was investigated.

[0163] Investigation of tableting performance: Investigate the state and stability of tablets.

[0164] Investigation of hard-shell capsules: Investigate the process feasibility and capsule stability.

[0165] Investigation of reconstitution performance: Investigate the dissolution rate and solution homogeneity.

[0166] Table 4. Results of investigation of liposome tableting performance

[0167]

[0168] Through the analysis of the data in Table 4, the following conclusions can be drawn:

[0169] High-purity phospholipids (such as soybean lecithin PC90 and above): Help improve the chemical stability of the product and ensure a high retention rate of vitamins in high-temperature accelerated stability tests. The vitamin retention rates in Examples 2, 3, 5, and 6 were all higher than 96%, while in Comparative Example 1 using low-purity phospholipids, the retention rates were slightly lower (VB6 95.8%, VB12 96.9%).

[0170] Adding appropriate wall materials: Not only improves the physical stability of liposomes, but also enhances their mechanical strength and pressure resistance during tableting, ensuring a higher vitamin retention rate. In Comparative Example 2 without adding wall materials, the vitamin retention rate decreased significantly to 93.9%.

[0171] Membrane layer stabilizers: Enhance the structural stability of liposomes, prevent the leakage of contents, and thus improve the vitamin retention rate. In Comparative Example 3 lacking membrane layer stabilizers, the vitamin retention rate decreased to 94.2%.

[0172] Antioxidants: They play a crucial role in preventing the oxidative degradation of vitamins. Especially in products containing easily oxidizable fat-soluble vitamins (such as VA, VD), adding antioxidants is essential. For example, in Comparative Example 4, the absence of antioxidants led to a significant decrease in the retention rates of VA and VD to 90.8% and 91.3% respectively.

[0173] Table 5. Results of the performance investigation of liposomal hard-shell capsules

[0174] Process Feasibility Stability Results (Retention Rate at 60°C for 20 Days) Example 2 Feasible 96.8％ Example 3 Feasible 98.2％ Example 5 Feasible <![CDATA[VB698.1%, VB 12 97.9%]]> Example 6 Feasible VA 97.3%, VD 98.4% Comparative Example 1 Feasible <![CDATA[VB696.1%, VB 12 96.6%]]> Comparative Example 2 Poor Fluidity, Process Not Feasible / Comparative Example 3 Feasible 97.1％ Comparative Example 4 Feasible VA 91.3%, VD 92.9%

[0175] Through the analysis of the data in Table 5, the following conclusions can be drawn:

[0176] Importance of wall materials: Adding appropriate wall materials not only improves the fluidity of liposomes, ensures the feasibility of the hard-shell capsule production process, but also enhances the physical stability of the product, preventing problems such as caking and poor fluidity.

[0177] Effect of phospholipid purity: High-purity phospholipids contribute to improving the chemical stability of the product, especially being more prominent in high-temperature accelerated stability tests. Although low-purity phospholipids can also achieve good results in some cases, considering other performance indicators comprehensively, high-purity phospholipids are a better choice.

[0178] Function of membrane stabilizers: Membrane stabilizers enhance the structural stability of liposomes, prevent the leakage of contents, and thus improve the retention rate of vitamins. The absence of membrane stabilizers will slightly reduce the stability of the product.

[0179] Cruciality of antioxidants: Antioxidants play a crucial role in preventing the oxidative degradation of vitamins. Especially in products containing easily oxidizable fat-soluble vitamins (such as VA, VD), adding antioxidants is essential to ensure long-term stability.

[0180] Table 6. Results of the performance investigation of liposomal reconstitution

[0181] Dispersion State Stand for 12 h Example 2 Rapid Dispersion, Uniform Dissolution Uniform Example 3 Rapid Dispersion, Uniform Dissolution Uniform Example 5 Rapid Dispersion, Uniform Dissolution Uniform Example 6 Rapid Dispersion, Uniform Dissolution Uniform Comparative Example 1 Slower Dispersion, Uniform Dissolution Uniform Comparative Example 2 Slower Dispersion, Uniform Dissolution Slight Flocculation Comparative Example 3 Rapid Dispersion, Uniform Dissolution Slight Flocculation Comparative Example 4 Rapid Dispersion, Uniform Dissolution Uniform

[0182] Through the analysis of the data in Table 6, the following conclusions can be drawn:

[0183] Importance of phospholipid purity: High-purity phospholipids (such as soybean lecithin PC90 and above) contribute to improving the dispersion speed and stability of liposomes, avoiding problems such as slow dispersion caused by unstable phospholipid membrane structures.

[0184] Key roles of wall materials and film layer stabilizers: Adding appropriate amounts of wall materials and film layer stabilizers can significantly improve the physical stability of liposomes and prevent them from flocculating during standing. This is very important for maintaining the long-term stability and consistency of the product. Influence of antioxidants: Although the influence of antioxidants on the reconstitution performance is relatively small, they play a key role in the accelerated stability test to ensure the efficient retention of vitamin components.

[0185] In summary, according to the investigation results of the application performance of liposomes, analyze the applicability of their tablet pressing application, hard shell capsule application, and solid beverage application.

[0186] Tablet pressing application: The liposomes of Examples 2, 3, 5, and 6 were tested by tablet pressing and their stability was investigated. The process was feasible and the stability passed. The liposome processes of Comparative Examples 1 to 4 were feasible, but the retention rates of the tablets prepared in Comparative Examples 2 and 3 decreased significantly after 20 days of acceleration. This was because without the protection of wall materials and film layer stabilizers, they were affected by mechanical forces during tablet pressing, which damaged their structure to a certain extent, resulting in a decrease in the vitamin retention rate.

[0187] Hard shell capsule application: All examples passed the test. The liposomes of Comparative Example 2 could not be applied to hard shell capsules because of their poor fluidity, which easily blocked the discharge port.

[0188] Solid beverage application: The liposomes of Examples 2, 3, 5, and 6 could be quickly dispersed and dissolved uniformly after reconstitution, with an average particle size below 310 nm, and remained uniform after standing for 12 h. However, the liposomes of Comparative Examples 1 and 2 dispersed slowly, the particle size increased, and slight flocculation occurred after standing for 12 h. The reason was also that wall materials and film layer stabilizers were not added, and the phospholipid membranes of the liposomes fused after dissolving in water, resulting in the generation of flocculation and making them unable to be well applied to solid beverages.

[0189] The above is only a preferred specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Those skilled in the art can easily think of changes or substitutions within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention. Therefore, the protection scope of the present invention should be subject to the protection scope of the claims.

Claims

1. A composite embedded vitamin liposome, characterized in that: It is prepared from the following raw materials in parts by weight: 0.1 to 80 parts of vitamins, 10 to 60 parts of phospholipids, 0.1 to 3 parts of antioxidants, 1 to 12 parts of film layer stabilizers, and 3 to 25 parts of wall materials.

2. The composite-embedded vitamin liposome according to claim 1, wherein: The vitamins described are water-soluble vitamins and / or fat-soluble vitamins.

3. The composite embedded vitamin liposome according to claim 1, wherein: The water-soluble vitamins are selected from one or more of vitamin C, vitamin B1, vitamin B2, vitamin B6, and vitamin B12; the fat-soluble vitamins are selected from one or more of vitamin A, vitamin D, vitamin E, and vitamin K.

4. The composite embedded vitamin liposome according to claim 1, characterized in that: The phospholipids are selected from one or more of soybean lecithin, sunflower lecithin, and rapeseed lecithin; The acetone-insoluble matter content in the phospholipids is greater than 60%; The phosphatidylcholine content in the phospholipids is greater than 50%.

5. The composite-embedded vitamin liposome according to claim 1, wherein: The film layer stabilizers include one or more of phytosterols, mono- and diglycerol fatty acid esters, fatty acids, and vegetable oils; The melting points of the mono- and diglycerol fatty acid esters, fatty acids, and vegetable oils are 40 to 90 °C.

6. The composite embedded vitamin liposome according to claim 1, characterized in that: The weight ratio of the film layer stabilizer to the phosphatidylcholine component in the phospholipids is 1:3 to 1:

15.

7. The composite embedded vitamin liposome according to claim 1, characterized in that: The phytosterols are selected from one or more of sitosterol, stigmasterol, brassicasterol, ergosterol, and microalgal sterol.

8. The composite embedded vitamin liposome according to claim 1, wherein: The antioxidants are selected from one or more of ascorbic acid, ascorbyl palmitate, mixed tocopherols, citrus flavonoids, proanthocyanidins, tea polyphenols, and carotenoids.

9. The composite-embedded vitamin liposome according to claim 1, wherein: The wall materials include one or more of modified starch, gum arabic, maltodextrin, resistant dextrin, oligo-maltose, and oligo-isomaltose.

10. The preparation method of the composite-embedded vitamin liposome according to claim 1, which comprises the following steps: S1. Preparation of the aqueous phase: Prepare an aqueous solution of water-soluble vitamins; S2. Preparation of the alcoholic phase: After anhydrous ethanol, phospholipids, and film layer stabilizers are fully mixed and dissolved, fat-soluble vitamins are added until completely dissolved; S3. Preparation of the wall material solution: Prepare an aqueous solution of the wall material; S4. Slowly inject the alcoholic phase into the aqueous phase, stir well to form a primary liposome dispersion solution, concentrate it under reduced pressure to obtain a concentrated solution, and slowly add it to the aqueous solution of the wall material obtained in S3, stir well to form a composite-embedded liposome dispersion solution; S5. Perform high-speed shearing, high-pressure homogenization, and drying on the composite-embedded liposome dispersion solution obtained in S4 to obtain the product.

11. According to the method described in claim 10, it is characterized in that: The solid content of the wall material solution in S3 is 20 to 60 wt%; The weight ratio of water to alcohol in S4 is (1 to 8):

1.

12. The application of the composite-embedded vitamin liposome according to claim 1 includes the production of tablet preparations, hard-shell capsules, and solid beverages.

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