Fat-soluble component nano-emulsion and preparation method thereof

Abstract

The invention relates to the technical field of preparation of health-care products, in particular to a fat-soluble component nano-emulsion and a preparation method thereof. The preparation method comprises the following steps: stirring and dissolving the antioxidant and the caprylic / capric triglyceride, adding the vitamin D3 and the retinyl palmitate, and stirring and dissolving to obtain an oil phase; stirring the starch sodium octenylsuccinate in purified water until the starch sodium octenylsuccinate is dissolved, adding glycerol, and uniformly stirring and mixing to obtain a water phase; adding a small amount of water phase into the oil phase, and stirring at high speed to obtain primary emulsion; adding the primary emulsion into the residual water phase, stirring at a high speed, shearing and homogenizing to obtain a nano embedding emulsion; adding citric acid, cane sugar and sodium ascorbate into purified water, adding the nano embedding emulsion, and uniformly stirring to obtain a concentrated solution; and diluting the concentrated solution and adjusting the pH value to obtain the fat-soluble component nano-emulsion. The fat-soluble component nano-emulsion provided by the invention has good safety; the product can be easily absorbed and utilized by a human body, and is convenient for people to quickly supplement vitamin A and vitamin D3 required by the body.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of health product preparation, in particular to a nanometer emulsion of fat-soluble ingredients and a preparation method thereof. BACKGROUND

[0002] With the improvement of living standards and the enhancement of health awareness, people's demand for health products is increasing. Vitamin A and vitamin D, as essential trace elements for the human body, play an important role in maintaining human health. Vitamin A helps maintain visual function, skin health, and promotes growth and development, while vitamin D has a positive effect on bone health, immune function, and cardiovascular health.

[0003] Since the human body cannot synthesize these two vitamins, it must meet the demand through external intake. The content of vitamin A and vitamin D in daily diet is limited, and is affected by many factors such as region, season, eating habits, etc., so it is often difficult for people to meet the body's demand for these two vitamins through diet. At the same time, with the acceleration of life pace, more and more people tend to choose convenient and fast eating habits, ignoring the importance of balanced diet, further reducing the intake of vitamin A and vitamin D.

[0004] Vitamin AD in the form of oral liquid is more easily absorbed by the human body, with high bioavailability, and the demand for vitamin AD oral liquid in the current population has increased significantly. Vitamin AD are both fat-soluble ingredients, but the current nanometer emulsion of fat-soluble ingredients has poor stability, and the content of vitamin A and vitamin D in the oral liquid will decrease over time, resulting in a decrease in health effects. Therefore, a nanometer emulsion of fat-soluble ingredients with high stability and convenient storage is needed. SUMMARY

[0005] The first object of the present application is to provide a nanometer emulsion of fat-soluble ingredients, which is easily absorbed and utilized by the human body, and facilitates people to quickly supplement the required vitamin A and vitamin D in the body.

[0006] The second object of the present application is to provide a preparation method of a nanometer emulsion of fat-soluble ingredients.

[0007] The preparation method of the nanometer emulsion of fat-soluble ingredients provided by the present application has the characteristics that it comprises the following steps: S1. Preparation of oil phase: antioxidant and caprylocaproyl glycerides are stirred and dissolved at 60-80℃, and after dissolution, cooled to 40-50℃, add vitamin D3 and stir until dissolved; then add palmitate retinyl ester, stir until dissolved, continue to stir to obtain the oil phase; S2. Preparation of aqueous phase: add sodium octenyl succinate starch into purified water, stir at 50-70℃ until dissolved, cool to room temperature and stand for defoaming, then add glycerol, stir to mix evenly, to obtain the aqueous phase; S3. Preparation of emulsion: under stirring, take a small amount of the aqueous phase obtained in step S2 and add into the oil phase obtained in step S1, then high-speed stirring to obtain the primary emulsion; add the primary emulsion into the remaining aqueous phase, high-speed stirring for 15-20 min, then shear homogenization to obtain the nano-embedded emulsion; S4. Preparation of concentrated solution: add citric acid, sucrose and sodium ascorbate into purified water, stir until dissolved, then add the nano-embedded emulsion prepared in step S3, stir evenly to obtain the concentrated solution; S5. Preparation of oral solution: dilute the concentrated solution obtained in step S4, and adjust the pH value to 4.0-6.0 to obtain the nano-emulsion of fat-soluble ingredients.

[0008] Preferably, the mass ratio of antioxidant, capryol 90, vitamin D3 and retinyl palmitate in step S1 is (1-5):(30-40):(0.1-1):(30-50); the mass ratio of purified water, sodium octenyl succinate starch and glycerol in step S2 is (5-100):(1-10):(1-5); the mass ratio between the total amount of oil phase and aqueous phase in step S3 for preparing the nano-embedded emulsion is 1:(10-20).

[0009] Preferably, the antioxidant includes any one of dibutylhydroxytoluene, butylated hydroxyanisole and alpha-tocopherol.

[0010] Preferably, the duration of standing for defoaming in step S2 is 4-6 hours.

[0011] Preferably, the specific steps of step S3 include: under stirring, slowly add a small amount of the aqueous phase obtained in step S2 into the oil phase obtained in step S1, high-speed stirring at 500-600 rpm for 15-25 min to obtain the primary emulsion; add the primary emulsion into the remaining aqueous phase, high-speed stirring at 550-700 rpm for 15-20 min, then high-speed shear homogenization at 7000-8000 rpm for 25-35 min to obtain the nano-embedded emulsion.

[0012] Preferably, the mass ratio of oil phase and aqueous phase in step S3 for preparing the primary emulsion is 1:1.

[0013] Preferably, the concentrated solution obtained in step S4 includes the following components in mass fraction: purified water 30-50 parts, citric acid 0.1-0.8 parts, sucrose 0.05-1.10 parts, sodium ascorbate 0.1-0.5 parts and nano-embedded emulsion 0.07-11.00 parts. The sucrose can be sucralose.

[0014] Preferably, the specific steps of the step S5 comprise: adding fructose glucose syrup, essence and the concentrated solution obtained in the step S4 into purified water, stirring and dispersing uniformly, then adding a pH adjuster to adjust the pH value to 4.0-6.0, and calibrating to a target volume to obtain a nanometer emulsion of fat-soluble ingredients. When diluting, purified water can be added according to the concentration requirement of the oral solution, and the stability of vitamin A and D in the oral solution will not be affected; the type of essence can be selected according to actual needs to adapt to the taste requirements of different people.

[0015] Preferably, the pH adjuster is a 10% sodium hydroxide solution.

[0016] The application further provides a nanometer emulsion of fat-soluble ingredients prepared by the preparation method.

[0017] Beneficial effects: The application provides a nanometer emulsion of fat-soluble ingredients, all the raw and auxiliary materials used are food-grade, and the nanometer emulsion has good safety; the particle size of the nanometer emulsion reaches the nanometer level, greatly increases the contact area with the human digestive and absorption organs, is easily absorbed and utilized by the human body, and facilitates people to quickly supplement vitamin A and vitamin D3 required by the human body.

[0018] The application further provides a preparation method of the nanometer emulsion of fat-soluble ingredients. When preparing an oil phase, adding an antioxidant can effectively improve the stability of the product; when preparing an aqueous phase, adding sodium starch octenyl succinate, which is an amphiphilic high molecular compound, can help to form a uniform aqueous phase due to its good emulsifying performance; in the process of preparing an emulsion, the oil phase is slowly added into the aqueous phase, and through shearing and stirring, the components of the oil phase are combined with the hydrophobic groups in the sodium starch octenyl succinate, and an oil-in-water emulsion is gradually formed. Under the conditions of high-speed shearing and homogenization, the particle size of the emulsion is reduced to below 1 micrometer, the embedding rate reaches 99%+, a stable nanometer oil-in-water dispersion system is formed, and the product can be ensured to have no phenomena such as precipitation, stratification and oil separation during long-term storage. When preparing a concentrated solution, adding citric acid can eliminate the possible metal ions in the emulsion, effectively avoid the catalytic oxidation of the metal ions on vitamin A and vitamin D3, and sodium ascorbate has strong reducing property and can synergistically antioxidize with vitamin A and vitamin D3. It can not only directly scavenge free radicals in the system, but also reduce the oxidized vitamin A and vitamin D3 to active forms, thereby improving the stability of the final product. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 A particle size distribution diagram of the nanometer emulsion of fat-soluble ingredients prepared in Example 1. DETAILED DESCRIPTION

[0020] It should be noted that the following detailed description is illustrative only, and is intended to provide further description in order to provide a thorough understanding of the present application. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the application pertains.

[0021] It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments in accordance with the present application. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that the terms "comprises" and / or "comprising," when used in this specification, specify the presence of stated features, steps, operations, elements, components, and / or groups thereof, but do not preclude the presence or addition of one or more other features, steps, operations, elements, components, and / or groups thereof.

[0022] The technical solutions of the present application will be described clearly and completely below in conjunction with the embodiments. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work fall within the scope of protection of the present application.

[0023] Embodiment 1 A preparation method of a nanometer emulsion of a fat-soluble ingredient, comprising the following steps: S1. Preparation of oil phase: take 3.0 g of antioxidant butyl hydroxy anisole, 35.0 g of caprylic capric glyceride, and dissolve under stirring in a 70°C water bath. After dissolution, cool to 40-50°C, then add 0.8 g of vitamin D3 under a 40°C water bath, and stir until dissolved. Then add 30.0 g of palmitate retinyl ester, stir until dissolved, and continue to stir in a water bath for 30 min to obtain the oil phase; S2. Preparation of water phase: add 0.5 kg of sodium octenyl succinate starch to 5.0 kg of purified water, and stir at a constant speed of 200 r / min at 60°C until dissolved. Cool to room temperature and stand for 4-6 hours to defoam (to fully defoam, the sodium octenyl succinate starch solution can be prepared one day in advance). Then add 0.3 kg of glycerol and stir to mix evenly to obtain the water phase; S3. Preparation of emulsion: under stirring, take 78.5 g of the water phase obtained in step S2 and add to the oil phase obtained in step S1, and stir at a high speed of 500 rpm for 20 min to obtain a primary emulsion. Add the primary emulsion to the remaining water phase, and stir at a high speed of 600 rpm for 18 min. Then use a SH digital constant speed shearing emulsifying stirrer to homogenize at a high speed of 8000 rpm for 30 min to obtain a nanometer-embedded emulsion; S4. Preparation of concentrate: Add 0.5 kg citric acid, 0.60 kg sucralose and 0.3 kg sodium ascorbate to 40.0 kg purified water, stir until dissolved, then add the nano-embedded emulsion prepared in step S3, stir evenly to obtain concentrate; S5. Preparation of oral liquid: Add 9.0 kg of fructose syrup, 0.30 kg of cantaloupe flavoring and the concentrated liquid obtained in step S4 to 900 kg of purified water. After stirring and dispersing evenly, add 10% sodium hydroxide solution as a pH adjuster to adjust the pH to 4.0-6.0. Then add an appropriate amount of purified water to make up to 1000 L to obtain a lipid-soluble nanoemulsion.

[0024] Example 2 A method for preparing a lipid-soluble nanoemulsion includes the following steps: S1. Preparation of the oil phase: Take 1.0 g of antioxidant dibutylhydroxytoluene and 30.0 g of caprylic / capric triglyceride, stir and dissolve them in a water bath at 60-80℃. After dissolution, cool to 40-50℃, then add 0.1 g of vitamin D3 in a water bath at 40℃ and stir until dissolved. Then add 40.0 g of retinyl palmitate and stir until dissolved. Continue stirring in a water bath for 30 min to obtain the oil phase. S2. Preparation of aqueous phase: Add 0.1 kg of sodium octenyl succinate starch to 0.50 kg of purified water, and stir at 50°C with a constant speed electric stirrer at 200 rpm until dissolved. Cool to room temperature and let stand to defoam for 4-6 hours (to fully defoam, the sodium octenyl succinate starch solution can be prepared one day in advance). Then add 0.1 kg of glycerol and stir to mix evenly to obtain the aqueous phase. S3. Preparation of emulsion: Under stirring, take 61.1g of the aqueous phase obtained in step S2 and add it to the oil phase obtained in step S1 according to the mass fraction. Stir at high speed of 600rpm for 15min to obtain the primary emulsion. Add the primary emulsion to the remaining aqueous phase and stir at high speed of 550rpm for 20min. Then use an SH digital display constant speed shear emulsification mixer to homogenize at high speed of 7000rpm for 35min to obtain the nano-embedded emulsion. S4. Preparation of concentrated solution: Add 0.1 kg citric acid, 0.05 kg sucralose and 0.1 kg sodium ascorbate to 30.0 kg purified water, stir until dissolved, then add the nano-embedded emulsion prepared in step S3, stir evenly to obtain concentrated solution; S5. Preparation of oral liquid: Add 8.0 kg of fructose syrup, 0.10 kg of cantaloupe flavoring and the concentrated liquid obtained in step S4 to 900 kg of purified water. After stirring and dispersing evenly, add 10% sodium hydroxide solution as a pH adjuster to adjust the pH to 4.0-6.0. Then add an appropriate amount of purified water to make up to 1000 L to obtain a lipid-soluble nanoemulsion.

[0025] Example 3 A method for preparing a lipid-soluble nanoemulsion includes the following steps: S1. Preparation of the oil phase: Take 5.0 g of antioxidant α-tocopherol and 40.0 g of caprylic / capric triglyceride, stir and dissolve them in a water bath at 60-80℃. After dissolution, cool to 40-50℃, then add 1.0 g of vitamin D3 in a water bath at 40℃ and stir until dissolved. Then add 50.0 g of retinyl palmitate and stir until dissolved. Continue stirring in a water bath for 30 min to obtain the oil phase. S2. Preparation of aqueous phase: Add 1.0 kg of sodium octenyl succinate starch to 10.0 kg of purified water, and stir at 70°C with a constant speed electric stirrer at 200 rpm until dissolved. Cool to room temperature and let stand to defoam for 4-6 hours (to fully defoam, the sodium octenyl succinate starch solution can be prepared one day in advance). Then add 0.5 kg of glycerol and stir to mix evenly to obtain the aqueous phase. S3. Preparation of emulsion: Under stirring, take 96g of the aqueous phase obtained in step S2 and add it to the oil phase obtained in step S1 according to the mass fraction. Stir at high speed of 600rpm for 25min to obtain the primary emulsion. Add the primary emulsion to the remaining aqueous phase and stir at high speed of 700rpm for 20min. Then use an SH digital display constant speed shear emulsification mixer to homogenize at high speed of 7500rpm for 25min to obtain the nano-embedded emulsion. S4. Preparation of concentrated solution: Add 0.8 kg citric acid, 1.10 kg sucralose and 0.5 kg sodium ascorbate to 50.0 kg purified water, stir until dissolved, then add the nano-embedded emulsion prepared in step S3, stir evenly to obtain concentrated solution; S5. Preparation of oral liquid: Add 10.0 kg of fructose syrup, 0.50 kg of cantaloupe flavoring and the concentrated liquid obtained in step S4 to 900 kg of purified water. After stirring and dispersing evenly, add 10% sodium hydroxide solution as a pH adjuster to adjust the pH to 4.0-6.0. Then add an appropriate amount of purified water to make up to 1000 L to obtain a lipid-soluble nanoemulsion.

[0026] Example 4 A method for preparing a lipid-soluble nanoemulsion includes the following steps: Steps S1, S2, S4, and S5 are the same as in Example 1; only step S3 is different. The specific steps of step S3 are as follows: S3. Emulsion Preparation: The oil phase obtained in step S1 is placed in an electrolytic cell and dispersed under an electric field with an electric field strength of 2-8 V / cm and a frequency of 20-600 Hz. Then, under stirring, 78.5 g of the aqueous phase obtained in step S2 is added to the dispersed oil phase, and the mixture is stirred at 500 rpm for 20 min to obtain a primary emulsion. The primary emulsion is added to the remaining aqueous phase and stirred at 600 rpm for 18 min. Then, the mixture is homogenized by high-speed shearing at 8000 rpm for 30 min using an SH digital display constant speed shearing mixer to obtain a nano-embedded emulsion. Oil droplets are subjected to an electric field force in an electric field. When the electric field strength reaches the range described in Example 4, the electric field force causes the oil droplets to deform and break into smaller droplets. These smaller droplets are more uniformly dispersed in the aqueous phase, increasing the contact area between the oil and aqueous phases, which is beneficial for the uniform dispersion of the oil phase in the aqueous phase. The electric field helps to form a charge layer on the surface of the oil droplets, generating electrostatic repulsion between the droplets, thereby reducing droplet aggregation and improving the stability of the nanoemulsion.

[0027] By adjusting parameters such as electric field strength and frequency, the particle size of nanoemulsions can be precisely controlled, allowing for better control of droplet size and distribution, resulting in a more uniform particle size distribution in the final nanoemulsion.

[0028] The above dispersion process is based on the principle of directional migration of ions under the action of an electric field. In the preparation of lipid-soluble nanoemulsions, by applying an external electric field, vitamin A, vitamin D3, and related functional components (such as antioxidants) can be charged and migrate directionally under the action of the electric field, thereby achieving uniform distribution and efficient encapsulation of the components and improving the quality and performance of the nanoemulsion.

[0029] Comparative Example 1 A method for preparing a lipid-soluble nanoemulsion includes the following steps: S1. Preparation of the oil phase: Take 5g of sodium EDTA-2 and 40.0g of caprylic / capric triglyceride, stir and dissolve them in a water bath at 60-80℃. After dissolving, cool to 40-50℃, then add 1.0g of vitamin D3 in a water bath at 40℃ and stir until dissolved. Then add 50.0g of retinyl palmitate and stir until dissolved. Continue stirring in a water bath for 30min to obtain the oil phase. S2. Preparation of aqueous phase: Add 0.5 kg of sodium octenyl succinate starch to 2 kg of purified water, and stir at 70°C with a constant speed electric stirrer at 200 rpm until dissolved. Cool to room temperature and let stand to defoam for 4-6 hours (to fully defoam, the sodium octenyl succinate starch solution can be prepared one day in advance). Then add 0.5 kg of glycerol and stir to mix evenly to obtain the aqueous phase. S3. Preparation of emulsion: Under stirring, take 96g of the aqueous phase obtained in step S2 and add it to the oil phase obtained in step S1 according to the mass fraction. Stir at high speed of 600 rpm for 25 min to obtain the primary emulsion. Add the primary emulsion to the remaining aqueous phase and stir at high speed of 700 rpm for 20 min. Then use an SH digital display constant speed shear emulsification mixer to homogenize at high speed of 7500 rpm for 25 min to obtain vitamin AD nano-embedded emulsion. S4. Preparation of concentrated solution: Add 0.8 kg citric acid, 1.10 kg sucralose and 0.5 kg sodium ascorbate to 50.0 kg purified water, stir until dissolved, then add the vitamin AD nano-embedded emulsion prepared in step S3, stir evenly to obtain concentrated solution; S5. Preparation of oral liquid: Add 10.0 kg of fructose syrup, 0.50 kg of cantaloupe flavoring and the concentrated liquid obtained in step S4 to 900 kg of purified water. After stirring and dispersing evenly, add 10% sodium hydroxide solution as a pH adjuster to adjust the pH to 4.0-6.0. Then add an appropriate amount of purified water to make up to 1000 L to obtain a lipid-soluble nanoemulsion.

[0030] Comparative Example 2 A method for preparing a lipid-soluble nanoemulsion includes the following steps: S1. Preparation of the oil phase: Take 10.0 g of ascorbyl palmitate and 40.0 g of caprylic / capric triglyceride, stir and dissolve them in a water bath at 60-80℃. After dissolving, cool to 40-50℃, and then add 1.0 g of vitamin D3 in a water bath at 40℃ and stir until dissolved. Then add 50.0 g of retinyl palmitate and stir until dissolved. Continue stirring in a water bath for 30 min to obtain the oil phase. S2. Preparation of aqueous phase: Add 0.5 kg of sodium octenyl succinate starch to 1.0 kg of purified water, and stir at 70°C with a constant speed electric stirrer at 200 rpm until dissolved. Cool to room temperature and let stand to defoam for 4-6 hours (to fully defoam, the sodium octenyl succinate starch solution can be prepared one day in advance). Then add 0.5 kg of glycerol and stir to mix evenly to obtain the aqueous phase. S3. Preparation of emulsion: Under stirring, take 96g of the aqueous phase obtained in step S2 and add it to the oil phase obtained in step S1 according to the mass fraction. Stir at high speed of 600 rpm for 25 min to obtain the primary emulsion. Add the primary emulsion to the remaining aqueous phase and stir at high speed of 700 rpm for 20 min. Then use an SH digital display constant speed shear emulsification mixer to homogenize at high speed of 7500 rpm for 25 min to obtain vitamin AD nano-embedded emulsion. S4. Preparation of concentrated solution: Add 0.8 kg citric acid, 1.10 kg sucralose and 0.5 kg sodium ascorbate to 50.0 kg purified water, stir until dissolved, then add the vitamin AD nano-embedded emulsion prepared in step S3, stir evenly to obtain concentrated solution; S5. Preparation of oral liquid: Add 10.0 kg of fructose syrup, 0.50 kg of cantaloupe flavoring and the concentrated liquid obtained in step S4 to 900 kg of purified water. After stirring and dispersing evenly, add 10% sodium hydroxide solution as a pH adjuster to adjust the pH to 4.0-6.0. Then add an appropriate amount of purified water to make up to 1000 L to obtain a lipid-soluble nanoemulsion.

[0031] Test case The particle size of the lipid-soluble nanoemulsion prepared in Example 1 was measured, and the test results are as follows: Figure 1 As shown, by Figure 1 It can be seen that the vitamin A and vitamin D particles in the lipid-soluble nanoemulsion prepared by this invention are small in size and have a narrow particle size distribution, with an average particle size of 36 nm. This indicates that the overall distribution of particles in the lipid-soluble nanoemulsion is relatively concentrated and the particle size is uniform.

[0032] Performance testing: The lipid-soluble nanoemulsion prepared in Example 1, as well as the lipid-soluble nanoemulsions prepared in Comparative Examples 1 and 2, were used as samples for performance testing.

[0033] 1. Water solubility test: The lipid-soluble nanoemulsion prepared in Example 1 can be rapidly dissolved in water, and the solution is clear, indicating that the lipid-soluble nanoemulsion has good water solubility.

[0034] 2. Stability testing: 2.1 Physical stability test 2.1.1 Centrifugation Experiment Nanoemulsions with adequate physical stability should not exhibit stratification under the experimental conditions of centrifugation at 4000 r / min for 15 min. 30 ml of the lipid-soluble nanoemulsions from Example 1, Comparative Example 1, and Comparative Example 2 were placed in centrifuge tubes and centrifuged at 4000 r / min for 15 min. The results are as follows: The product of Example 1 showed no stratification after centrifugation, the solution was clear, the transmittance was 95%, and the stability met the requirements. The product of Comparative Example 1 showed a 0.3 cm layer at the bottom of the test tube after centrifugation, and the upper layer was slightly turbid. The product of Comparative Example 2 showed no obvious stratification after centrifugation, but the solution was slightly milky (transmittance 82%). It can be seen that the product of Example 1 has significantly better stability and dispersibility than Comparative Example 1 after centrifugation.

[0035] 2.1.2 Shaking Test In Example 1, five groups of 30 mL lipid-soluble nanoemulsions were sealed in 50 mL test tubes and shaken at 100 rpm in a 30 °C constant temperature water bath shaker. Samples were taken at 12 h, 24 h, 48 h, and 72 h, and the centrifugal stability parameters (Ke) of the lipid-soluble nanoemulsions at each sampling time were measured. The test results are shown in Table 1.

[0036] Table 1

[0037] As shown in Table 1, the lipid-soluble nanoemulsion prepared in Example 1 still exhibits good stability under continuous shaking, and shaking does not reduce its stability.

[0038] The products of Comparative Example 1 and Comparative Example 2 were subjected to shaking tests using the parameters described above, and samples were taken at their initial state and after 72 hours to determine the centrifugal stability parameter (Ke). The results showed that the Ke value of the product of Comparative Example 1 was 0.315 at 0 hours and 0.482 at 72 hours, an increase of 53%; the Ke value of the product of Comparative Example 2 was 0.313 at 0 hours and 0.395 at 72 hours, an increase of 26.2%. In contrast, the Ke value of the product of Example 1 increased by only 6.1% at 72 hours compared to its initial state, demonstrating significantly better stability than the products of Comparative Example 1 and Comparative Example 2.

[0039] 2.1.3 Temperature Test In Example 1, three groups of 30 mL lipid-soluble nanoemulsions were sealed in glass bottles and placed at 40℃, 60℃, and 70℃ for 10 days, respectively. The particle size and distribution of the lipid-soluble nanoemulsions were examined at 0, 5, and 10 days. The average particle size test results are shown in Table 2.

[0040] Table 2

[0041] As shown in Table 2, the particle size of the lipid-soluble component nanoemulsion prepared in Example 1 did not change significantly at different temperatures.

[0042] The products of Comparative Example 1 and Comparative Example 2 were subjected to temperature experiments using the above method. They were placed at 70°C for 10 days, and the initial particle size and the particle size at 10 days were measured.

[0043] After measurement, the initial particle size of the product of Comparative Example 1 was 37 nm, and the particle size was 62 nm after 10 days, with an increase of 67.6%; the initial particle size of the product of Comparative Example 2 was 36 nm, and the particle size was 51 nm after 10 days, with an increase of 41.7%. However, the particle size increase of the product of Example 1 under the same test conditions was only 27.8%, and its particle size stability was much better than that of Comparative Example 1 and Comparative Example 2.

[0044] 2.1.4 Accelerated Testing The lipid-soluble component nanoemulsion prepared in Example 1 was placed at 37°C for 6 months. The physical properties of the lipid-soluble component nanoemulsion at 0, 1, 2, 3 and 6 months were investigated. The average particle size test results are shown in Table 3.

[0045] Table 3

[0046] Accelerated testing demonstrated that the particle size of the lipid-soluble nanoemulsion prepared in Example 1 did not change significantly at room temperature (30°C) and could be stored for a long time.

[0047] The products of Comparative Example 1 and Comparative Example 2 were subjected to accelerated testing using the above method to investigate the physical properties of the lipid-soluble component nanoemulsion at 0 months and 6 months.

[0048] The initial particle size of the product in Comparative Example 1 was 37 nm, and after 6 months the particle size reached 75 nm, an increase of 102.7%. The initial particle size of the product in Comparative Example 2 was 36 nm, and after 6 months the particle size reached 58 nm, an increase of 61.1%. In contrast, the particle size of the product in Example 1 was 45 nm after 6 months, with an increase of only 25.0%, indicating higher stability and ease of long-term storage.

[0049] 2.2 Chemical stability test 2.2.1 High Temperature Test The fat-soluble nanoemulsion prepared in Example 1 was filled into a sodium-calcium glass bottle and sealed. It was placed at 40°C, 60°C and 70°C for 10 days respectively. The changes in the content of the effective components vitamin A and vitamin D3 in the fat-soluble nanoemulsion were investigated at 0 days, 5 days and 10 days respectively. The test results are shown in Table 4.

[0050] Table 4

[0051] As shown in Table 4, the fat-soluble nanoemulsion prepared by this invention maintains stable levels of vitamin A and vitamin D3 when stored at the above-mentioned temperature for an extended period, ensuring the effectiveness of the components during storage.

[0052] The fat-soluble nanoemulsions prepared in Comparative Examples 1 and 2 were filled into sodium-calcium glass bottles and sealed. They were placed at 70°C for 10 days, and the changes in the content of the effective components vitamin A and vitamin D3 in the fat-soluble nanoemulsions were investigated at 0 days and 10 days, respectively.

[0053] In Comparative Example 1, the vitamin A content decreased from the initial 119.7 ug / 10 ml to 114.2 ug / 10 ml, a decrease of 4.6%, and the vitamin D3 content decreased from the initial 6.67 ug / 10 ml to 6.28 ug / 10 ml, a decrease of 5.8%. In Comparative Example 2, the vitamin A content decreased from the initial 119.6 ug / 10 ml to 116.3 ug / 10 ml, a decrease of 2.8%, and the vitamin D3 content decreased from the initial 6.66 ug / 10 ml to 6.41 ug / 10 ml, a decrease of 3.8%. In Example 1, the decrease in vitamin A was 1.4% and the decrease in vitamin D3 was 2.5% during the same period. Therefore, the degradation rate of the active ingredients in the products of Comparative Example 1 and Comparative Example 2 is much higher than that in Example 1, which is not conducive to long-term storage.

[0054] 2.2.2 Accelerated Testing The fat-soluble component nanoemulsion prepared in Example 1 was placed at 37°C for 6 months, and the changes in vitamin A and vitamin D3 content at 0, 1, 2, 3 and 6 months were investigated. The experimental results are shown in Table 5.

[0055] Table 5

[0056] As shown in Table 5, the content of the active ingredients vitamin A and vitamin D3 in the fat-soluble nanoemulsion remained relatively stable after long-term storage at 37°C.

[0057] The fat-soluble nanoemulsions prepared in Comparative Examples 1 and 2 were stored at 37°C for 6 months, and the changes in vitamin A and vitamin D3 content at 0 and 6 months were investigated. In Comparative Example 1, the vitamin A content decreased from the initial 119.7 ug / 10 ml to 110.3 ug / 10 ml, a decrease of 7.9%, and the vitamin D3 content decreased from the initial 6.67 ug / 10 ml to 6.05 ug / 10 ml, a decrease of 9.3%. In Comparative Example 2, the vitamin A content decreased from the initial 119.6 ug / 10 ml to 113.5 ug / 10 ml, a decrease of 5.1%, and the vitamin D3 content decreased from the initial 6.66 ug / 10 ml to 6.27 ug / 10 ml, a decrease of 5.9%. In Example 1, the decrease in vitamin A was 3.1% and the decrease in vitamin D3 was 3.9% during the same period. Therefore, the degradation rate of the active ingredients in the products of Comparative Examples 1 and 2 was much higher than that in Example 1, which is not conducive to long-term storage.

[0058] In summary, the antioxidants butylated hydroxytoluene / butylated hydroxyanisole / α-tocopherol used in this invention can directly scavenge free radicals in the system and are highly compatible with both the oil phase (glyceryl caprylate) and the aqueous phase (sodium octenyl succinate starch). This not only protects VA / VD3 from oxidation but also maintains the dispersion stability of the nanoparticles. Therefore, the physical and chemical stability of the lipid-soluble nanoemulsion prepared by this invention is far superior to that of Comparative Examples 1 and 2, which is beneficial for long-term storage.

[0059] 3. Safety test 3.1 Acute oral toxicity test The experiment was conducted using a controlled dose method. Twenty SD rats, half male and half female, with females weighing 195.7–212.2 g and males weighing 189.9–209.1 g, were used. They were fasted for 16 hours but had unrestricted water intake. A single dose group was established at 10.0 g / kg bw. 50.0 g of the test sample was diluted with pure water to 100 mL to prepare a test solution with a concentration of 500 mg / mL. The test solution was administered orally at a dose of 20 mL / kg bw. After gavage, the animals were kept fasted for another 4 hours and observed for 14 consecutive days. Symptoms of poisoning and mortality were recorded. Animals were weighed at the start of the experiment, on day 7, and at the end of the experiment. After the experiment, the animals were euthanized and subjected to gross necropsy.

[0060] According to GB15193.3-2014 "National Food Safety Standard - Acute Oral Toxicity Test", the LD50 was assessed as follows: 50 >10.0g / kg bw, which is practically non-toxic.

[0061] 3.2.28-day oral toxicity test One hundred weaned SD rats, half male and half female, were selected. Females weighed 74.8–100.0 g, and males weighed 80.0–100.0 g. The test substance was administered via feeding. Preliminary results indicated that the maximum tolerable concentration of the test substance incorporation in the animals was 10.0%. The high-dose group was set at 10.0 g / kg bw / d, with a 2-fold interval design. The medium- and low-dose groups were set at 5.0 mg / kg bw / d and 2.5 mg / kg bw / d (equivalent to 25, 50, and 100 times the proposed human dose, respectively). A control group was also included. A convalescent control satellite group and a high-dose satellite group were also added. Each experimental group consisted of 20 animals, and each satellite group consisted of 10 animals, with half males and half females. The animals were fed continuously for 28 days.

[0062] According to GB15193.22-2014 "National Food Safety Standard 28-Day Oral Toxicity Test", the results were evaluated. Under the experimental conditions, the No Observed Adverse Effect Level (NOAEL) for the tested samples in the 28-day oral toxicity test for female and male SD rats was 12.95 g / kg bw / d and 12.83 g / kg bw / do, respectively.

[0063] The above tests demonstrate that the fat-soluble nanoemulsion provided by this invention has high safety and meets national food safety requirements.

[0064] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for preparing a nanoemulsion of a fat-soluble component, characterized in that, Includes the following steps: S1. Preparation of the oil phase: Dissolve the antioxidant and caprylic / capric triglyceride at 60-80℃, cool to 40-50℃ after dissolution, add vitamin D3 and stir until dissolved; then add retinyl palmitate and stir until dissolved, continue stirring to obtain the oil phase; S2. Preparation of aqueous phase: Add sodium octenyl succinate starch to purified water, stir at 50-70℃ until dissolved, cool to room temperature and let stand to defoam, then add glycerol, stir and mix evenly to obtain aqueous phase; S3. Preparation of emulsion: Under stirring, take a small amount of the aqueous phase obtained in step S2 and add it to the oil phase obtained in step S1, and then stir at high speed to obtain the initial emulsion; add the initial emulsion to the remaining aqueous phase, stir at high speed for 15-20 min, and then shear homogenize to obtain the nano-embedded emulsion. S4. Preparation of concentrate: Add citric acid, sucrose and sodium ascorbate to purified water and stir until dissolved. Then add the nano-embedded emulsion prepared in step S3 and stir evenly to obtain concentrate. S5. Preparation of oral solution: Dilute the concentrated solution obtained in step S4 and adjust the pH value to 4.0-6.0 to obtain a lipid-soluble nanoemulsion.

2. The method for preparing the lipid-soluble component nanoemulsion according to claim 1, characterized in that, In step S1, the mass ratio of antioxidant, caprylic / capric triglyceride, vitamin D3 and retinyl palmitate is (1-5):(30-40):(0.1-1):(30-50); in step S2, the mass ratio of purified water, sodium octenyl succinate starch and glycerol is (5-100):(1-10):(1-5); in step S3, the mass ratio between the total amount of oil phase and water phase when preparing the nano-embedded emulsion is 1:(10-20).

3. The method for preparing the lipid-soluble component nanoemulsion according to claim 1, characterized in that, Antioxidants include any one of butylated hydroxytoluene, butylated hydroxyanisole, and α-tocopherol.

4. The method for preparing the lipid-soluble component nanoemulsion according to claim 1, characterized in that, The duration of the static defoaming process in step S2 is 4-6 hours.

5. The method for preparing the lipid-soluble component nanoemulsion according to claim 1, characterized in that, The specific steps of step S3 include: under stirring, taking a small amount of the aqueous phase obtained in step S2 and slowly adding it to the oil phase obtained in step S1, and stirring at high speed at 500-600 rpm for 15-25 min to obtain a primary emulsion; adding the primary emulsion to the remaining aqueous phase, stirring at high speed at 550-700 rpm for 15-20 min, and then homogenizing at high speed at 7000-8000 rpm for 25-35 min to obtain a nano-embedded emulsion.

6. The method for preparing the lipid-soluble component nanoemulsion according to claim 1, characterized in that, In step S3, when preparing the colostrum, the mass ratio of the oil phase to the water phase is 1:

1.

7. The method for preparing the lipid-soluble component nanoemulsion according to claim 1, characterized in that, The concentrate obtained in step S4 comprises the following components in parts by mass: 30-50 parts purified water, 0.1-0.8 parts citric acid, 0.05-1.10 parts sucrose, 0.1-0.5 parts sodium ascorbate, and 0.07-11.00 parts nano-embedding emulsion.

8. The method for preparing the lipid-soluble component nanoemulsion according to claim 1, characterized in that, The specific steps of step S5 include: adding fructose syrup, flavoring and the concentrate obtained in step S4 to purified water, stirring and dispersing evenly, adding a pH adjuster to adjust the pH to 4.0-6.0, and obtaining a lipid-soluble nanoemulsion.

9. The method for preparing the lipid-soluble component nanoemulsion according to claim 1, characterized in that, The pH adjuster is a 10% sodium hydroxide solution.

10. A lipid-soluble nanoemulsion prepared by any one of the preparation methods according to claims 1-9.

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