Preparation method and application of vitamin D dry powder raw material wrapped by nano fat emulsion

Abstract

The present application relates to a method and application of preparing a vitamin D dry powder raw material wrapped by a nano fat emulsion in the technical field of vitamin D raw material preparation, and it comprises the following steps: weighing the preparation material, the preparation material includes the following components and their mass percentages: fat-soluble vitamin D or vitamin D analog 0.5-20%, solubilizing component 1-5%, lecithin packaging material 1.5-10%, auxiliary packaging material 0.1-1%, antioxidant 0.01-0.05% and ion complexing agent 0.01-0.02%, and the remainder is excipient; the solubilizing component, fat-soluble vitamin D or vitamin D analog, lecithin packaging material, auxiliary packaging material, antioxidant and ion complexing agent are mixed to form colostrum; colostrum is homogenized by high pressure to a particle size of less than 500nm to obtain a fat emulsion vitamin D liquid; excipients are added to the fat emulsion vitamin D liquid, stirring and dissolving after filtering through a filter membrane, drying, and then crushing and sieving to obtain a nano fat emulsion wrapped vitamin D dry powder raw material. The present application can improve the stability and bioavailability of vitamin D raw materials.

Description

Technical Field

[0001] The present application relates to the field of vitamin D raw material preparation technology, and in particular to a preparation method and application of a vitamin D dry powder raw material encapsulated by a nano fat emulsion. Background Art

[0002] Vitamin D is a fat-soluble vitamin. The four-ring structure of vitamin D compounds is the same as the parent nucleus, but the side chains are different. There are at least 10 known vitamin D compounds, and the most important ones in clinical use are vitamin D2 and vitamin D3. Vitamin D has been favored by the world for its significant health and medical effects, but its promotion and application urgently need to solve the following three main problems: solubility, stability and bioavailability.

[0003] First, vitamin D is insoluble in water, a property that greatly limits its application in the pharmaceutical and food industries. Second, vitamin D is easily oxidized and decomposed by photocatalysis, especially under acidic conditions, and is very easy to degrade, resulting in a decrease in its content in the product. Third, because vitamin D is insoluble in water, it is not easily absorbed as an oral medicine or food, and its bioavailability is low and unstable, causing great trouble to clinical medication.

[0004] In order to solve the above-mentioned problems in the preparation process of vitamin D raw materials, the following existing technologies have emerged: it is reported that cyclodextrin encapsulation can improve the stability and water solubility of vitamin D; patents with announcement numbers CN113546062A and CN105311037B propose that dissolving vitamin D in vegetable oil can improve its stability; patent with announcement number CN104434780A proposes to form an emulsion through a surfactant, and this emulsion preparation increases the stability of vitamin D; patent with announcement number CN111346071A proposes that preparations prepared using polymers such as cellulose as excipients can stabilize and alleviate the degradation of vitamin D.

[0005] There are also reports that sodium starch octenyl succinate is used to encapsulate vitamin D to improve its stability during the preparation process and shelf life of solid preparations. However, due to the high brittleness of the encapsulated particles, they are easily broken during the mixing process of preparation, and the vitamin D without the protection of the polymer material will degrade quickly. In addition, the molecular weight, degree of substitution, and octenylation of the sodium starch octenyl succinate polymer encapsulating vitamin D are difficult to control, which is bound to cause great difficulty in controlling the quality of the final preparation drug. Therefore, this polymer excipient has not yet been approved for use by drug review agencies in various countries.

[0006] Moreover, most of the methods described above use excipients and additives in formulations and processes to improve stability, rather than protecting the source of raw materials. The vegetable oil dissolution method can stabilize vitamins to a certain extent, but it cannot help improve bioavailability, and it has a poor taste when taken. It can only be available in the form of oil-soluble soft capsules or drop pills, which limits the use of more dosage forms. Cyclodextrin technology is prohibited in the use of injections because it can cause serious side effects such as DNA mutations, and the drug loading is limited. Summary of the invention

[0007] In order to improve the stability and bioavailability of vitamin D raw materials, the present application provides a method for preparing a vitamin D dry powder raw material encapsulated by nano fat emulsion.

[0008] In the first aspect, the present application provides a method for preparing a nano fat emulsion-encapsulated vitamin D dry powder raw material using the following technical solution:

[0009] A method for preparing a vitamin D dry powder raw material wrapped by nano fat emulsion comprises the following steps:

[0010] Weighing the preparation material, the preparation material includes the following components and their mass percentages: 0.5-20% of fat-soluble vitamin D or vitamin D analogs, 1-5% of solubilizing component, 1.5-10% of lecithin packaging material, 0.1-1% of auxiliary packaging material, 0.01-0.05% of antioxidant, 0.01-0.02% of ion complexing agent, and the balance is excipient;

[0011] The solubilizing component, fat-soluble vitamin D or vitamin D analog, lecithin packaging material, auxiliary packaging material, antioxidant and ion complexing agent are mixed to prepare colostrum; the colostrum is homogenized by high pressure until the particle size is less than 500 nm to obtain fat emulsion vitamin D liquid;

[0012] Add excipients to the fat emulsion vitamin D liquid, stir and dissolve, filter through a filter membrane, dry, and then grind and sieve to obtain the vitamin D dry powder raw material wrapped in nano fat emulsion.

[0013] By adopting the above technical solution, fat emulsion technology is used to encapsulate fat-soluble vitamin D in fat emulsion droplets with a particle size of less than 500nm. The fat emulsion encapsulation can completely isolate vitamin D from the outside world, effectively avoiding the hydrolysis, enzymatic hydrolysis and acid-catalyzed decomposition of vitamin D, and also slowing down the degree of thermal decomposition and photodecomposition, so as to effectively improve the stability of the vitamin D raw material.

[0014] Nano fat encapsulates the poorly soluble compound vitamin D in the phospholipid-encapsulated solubilizing component droplets, which has a significant solubilizing effect and effectively improves the solubility of the vitamin D raw material. In addition, the fat emulsion obtained by the above method has a structure similar to that of biological membranes and chyle and similar basic components, good safety, good compatibility with biological membranes, and can promote the absorption of vitamin D through the mucosa; oral nano fat emulsion can also increase the adsorption of its encapsulated vitamin D in intestinal epithelial cells and prolong the absorption time, thereby effectively improving the bioavailability of vitamin D raw materials. This also makes it possible for the fat emulsion-encapsulated vitamin D to be directly taken orally as a raw material, or to be made into a preparation in combination with other excipients, and it is relatively easy to use in processed foods such as medicines, health products and beverages, which can not only meet people's demand for high-quality nutrients, but also fill the gap in the domestic food market.

[0015] At the same time, due to the dense arrangement and packaging of the phospholipid membrane formed by the lecithin packaging material and the relatively soft characteristics of the dry powder, it can effectively alleviate the damage caused by the friction of the powder during extrusion and mixing of fat emulsion vitamin D dry powder in the solid preparation preparation process, and the fat emulsion packaging effectively avoids the direct physical contact between vitamin D and the surface of the solid preparation auxiliary powder, avoiding the probability and degree of surface catalysis initiating and aggravating the degradation of vitamin D, and further improving the stability of the vitamin D raw material during the preparation process. The invention has a simple process, good safety of the packaging material and each auxiliary material, and controllable quality, and is suitable for large-scale industrial production.

[0016] Optionally, the lecithin packaging material includes one or a combination of egg yolk lecithin, soybean lecithin, hydrogenated egg yolk lecithin, hydrogenated soybean lecithin, artificial synthetic lecithin and sphingomyelin.

[0017] Optionally, the preparation of colostrum comprises the following steps:

[0018] The solubilizing component, fat-soluble vitamin D or vitamin D analogs, lecithin packaging material, auxiliary packaging material and antioxidant are mixed, heated and stirred to dissolve, and an oil phase is obtained; the ion complexing agent and water are mixed, and heated to obtain an aqueous phase; the oil phase is slowly added to the aqueous phase, the temperature of the aqueous phase is controlled at 45-55° C., high-speed shearing is performed, the addition time of the oil phase is controlled at 15-30 minutes, and stirring is continued for 20-30 minutes after mixing to obtain colostrum.

[0019] Optionally, the preparation of colostrum comprises the following steps:

[0020] The solubilizing component, fat-soluble vitamin D or vitamin D analogs, lecithin packaging material, auxiliary packaging material and antioxidant are dissolved in an organic mixed solvent, heated and stirred to dissolve, and the organic mixed solvent is removed by rotary evaporation to form a uniform phospholipid vitamin D film, and an appropriate amount of deionized water is added, and heated and stirred to hydrate and disperse the phospholipid vitamin D film to obtain colostrum.

[0021] Optionally, the preparation of colostrum comprises the following steps:

[0022] The solubilizing component, fat-soluble vitamin D or vitamin D analogs, lecithin packaging material, auxiliary packaging material and antioxidant are added to anhydrous ethanol, heated and stirred to dissolve to obtain a mixed solution, and then the mixed solution is slowly added to the ion complexing agent and deionized water in high-speed shearing, and stirring is continued for 10-30 minutes to obtain colostrum.

[0023] Optionally, a coating stabilizer accounting for 2-4% by weight of the colostrum is added and then high-pressure homogenization is performed, wherein the coating stabilizer is a gel prepared from triglyceride, sturgeon skin gelatin and hydroxyapatite in a mass ratio of 1:(0.4-0.6):(0.04-0.08).

[0024] By adopting the above technical solution, sturgeon skin gelatin has a strong gel strength, a clear microscopic network structure and relatively uniform voids. Hydroxyapatite is a porous material with high adsorption and carrying capacity. Sturgeon skin gelatin is compounded with hydroxyapatite, and sturgeon skin gelatin and hydroxyapatite are cross-linked and bonded to each other, forming a stable structure with high cross-linking density, softness and certain support strength on the surface of fat emulsion particles to prevent phospholipids from oxidative hydrolysis in aqueous solution and cause vitamin D leakage; and it can eliminate the physical contact such as friction and extrusion between the fat emulsion and the surface of the excipient powder to a certain extent, reducing the possibility of deterioration of the fat emulsion encapsulation effect; and it can increase the loading amount of the excipient on the fat emulsion vitamin D solution, thereby increasing the amount of vitamin D embedded in the raw material dry powder. Since hydroxyapatite itself has good biocompatibility and biodegradability, after the excipient is re-dissolved, the stable structure on the surface of the fat emulsion particles can be degraded and decomposed in the human body to release vitamin D; and compounding with triglycerides can further promote the degradation and decomposition of hydroxyapatite after entering the human body, while improving the stability of the vitamin D raw material, it also ensures the bioavailability and release efficiency of the vitamin D raw material.

[0025] Optionally, the method for preparing the coating stabilizer comprises the following steps:

[0026] The carbon triglyceride and sturgeon skin gelatin are added to deionized water, heated to 50-60° C., stirred and dissolved, and then a 0.3 g / L hydroxyapatite aqueous suspension is added under high-speed stirring, and the temperature is continued to be raised to 70-80° C., kept for reaction for 1-2 hours, then cooled to 25° C., stirred for 1-2 hours, and citric acid is added to adjust the pH to neutral to obtain a coating stabilizer.

[0027] Optionally, the excipient includes one or a combination of mannitol, sorbitol, polyethylene glycol, lactose, glucose, maltodextrin, dextran and trehalose.

[0028] By adopting the above technical solution and using the above excipients and coating stabilizers in combination, the stability of the vitamin D raw material and the encapsulation amount of vitamin D can be further effectively improved.

[0029] Optionally, the solubilizing component includes one or a combination of soybean oil, corn oil, peanut oil, sunflower seed oil, rapeseed oil, olive oil, pine oil, tea seed oil, galangal oil, ganoderma lucidum spore oil and sea buckthorn oil.

[0030] Optionally, the particle size of the fat emulsion in the fat emulsion vitamin D solution is 50-300nm.

[0031] By adopting the above technical solution, the fat emulsion with the above particle size can have a relatively higher vitamin D embedding amount in the subsequent raw material dry powder while having better solubility, stability and bioavailability.

[0032] Optionally, the homogenization pressure of the high-pressure homogenization is 95-110 MPa, and the number of homogenization times is 6-8 times.

[0033] In the second aspect, the present application provides a nano fat emulsion-encapsulated vitamin D dry powder raw material using the following technical solution:

[0034] A vitamin D dry powder raw material wrapped in nano fat emulsion is prepared by the preparation method of the vitamin D dry powder raw material wrapped in nano fat emulsion.

[0035] In the third aspect, the application of a vitamin D dry powder raw material encapsulated by a nano fat emulsion provided in the present application adopts the following technical solution:

[0036] The invention discloses an application of a vitamin D dry powder raw material wrapped in a nano fat emulsion, including solid preparations, oral liquid preparations and injection preparations. The application forms in the solid preparations include ordinary tablets, capsules, oral disintegrating tablets, granules and instant powders. The application forms in the oral liquid preparations include solutions, suspensions and drops. The application forms in the injection preparations include injections and dry powder injections.

[0037] By adopting the above technical solution, fat emulsion vitamin D dry powder can be directly prepared into solid preparations for application. Since fat emulsion vitamin D dry powder can still maintain nanometer-scale particle size after reconstitution, it solves the problem of dissolving fat-soluble vitamins and avoids the use of surfactants with side effects, and the obtained injection is safer; and the oral preparation prepared from fat emulsion vitamin D dry powder has good stability and improves the bioavailability of vitamin D when used as an oral liquid preparation.

[0038] In summary, the present application includes at least one of the following beneficial technical effects:

[0039] 1. The fat emulsion technology is used to encapsulate fat-soluble vitamin D in fat emulsion droplets with a particle size of less than 500nm. The fat emulsion encapsulation can completely isolate vitamin D from the outside world, effectively avoiding the hydrolysis, enzymatic hydrolysis and acid-catalyzed decomposition of vitamin D, and also slowing down the degree of thermal decomposition and photodecomposition, so as to effectively improve the stability of vitamin D raw materials;

[0040] 2. The nano-fat encapsulated insoluble compound vitamin D has a significant solubilizing effect in the solubilizing component droplets wrapped by lecithin, which effectively improves the solubility of the vitamin D raw material; and the fat emulsion obtained by the above method has a structure similar to that of biological membrane and chyle and similar basic components, good safety, and good compatibility with biological membrane, which can promote the absorption of vitamin D through the mucosa; oral nano-fat emulsion can also increase the adsorption of the encapsulated vitamin D in the intestinal epithelial cells and prolong the absorption time, thereby effectively improving the bioavailability of the vitamin D raw material;

[0041] 3. Due to the dense arrangement and wrapping of the phospholipid membrane formed by the lecithin packaging material and the relatively soft characteristics of the dry powder, it can effectively alleviate the damage caused by powder friction during extrusion and mixing of fat emulsion vitamin D dry powder in the solid preparation process, avoid the probability and degree of surface catalysis initiating and aggravating vitamin D degradation, and further improve the stability of vitamin D raw materials in the preparation process;

[0042] 4. Sturgeon skin gelatin and hydroxyapatite are cross-linked to form a stable structure with high cross-linking density, softness and certain supporting strength on the surface of fat emulsion particles, so as to prevent phospholipids from oxidative hydrolysis in aqueous solution and cause vitamin D leakage; and can eliminate physical contact such as friction and extrusion between the fat emulsion and the surface of excipient powder to a certain extent, and reduce the possibility of deterioration of fat emulsion encapsulation effect; and can increase the loading amount of excipient on fat emulsion vitamin D solution, thereby increasing the vitamin D embedding amount in the raw material dry powder; and the compounding of hydroxyapatite and triglyceride can further promote the degradation and decomposition of hydroxyapatite after entering the human body, while improving the stability of vitamin D raw material, it also ensures the bioavailability and release efficiency of vitamin D raw material. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1 It is a stability comparison chart of Example 3 of the present application and Comparative Examples 1-2.

[0044] Figure 2 It is a stability comparison chart of long-term sample retention inspection of Example 3 and Comparative Example 1 of the present application. DETAILED DESCRIPTION

[0045] The embodiments of the present invention will be described in detail below in conjunction with examples. However, those skilled in the art will understand that the following examples are only used to illustrate the present invention and should not be construed as limiting the scope of the present invention. The specific conditions not specified in the examples are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used without indicating the manufacturer are all conventional products that can be purchased commercially.

[0046] Example Part I: Different colostrum preparation methods

[0047] Embodiment 1:

[0048] A vitamin D dry powder raw material wrapped in nano fat emulsion, the preparation method of which comprises the following steps:

[0049] The preparation material is weighed with a total of 500 g, and the preparation material includes the following components and their mass percentages:

[0050] Vitamin D3 5%, solubilizing component 2%, lecithin packaging material 3%, auxiliary packaging material 0.5%, antioxidant 0.03%, ion complexing agent 0.03%, and the balance is excipients;

[0051] The dissolving component is soybean oil, the lecithin packaging material is soybean lecithin, the auxiliary packaging material is oleic acid, the antioxidant is vitamin E, the ion complexing agent is EDTA, and the excipient is lactose.

[0052] Add the above-mentioned amounts of soybean oil, vitamin D3, soybean lecithin, oleic acid and vitamin E into a reactor, heat to 65°C under nitrogen protection, and stir to completely dissolve the materials to obtain an oil phase solution; add 800-850 mL of deionized water into the reactor, add the above-mentioned amount of EDTA, adjust the pH of the solution to 8.5-9.5, and heat to 50°C to obtain an aqueous phase solution; under sufficient nitrogen protection, slowly add the prepared oil phase solution into the aqueous phase solution kettle, control the temperature of the aqueous phase kettle at 50°C, high-speed shearing, the shear speed is 8000 r / min, the adding time is controlled at 25 min, and stirring is continued for 30 min after mixing to obtain colostrum.

[0053] The colostrum prepared above was homogenized 7 times by a high-pressure homogenizer with a pressure of 100 MPa. The internal control index was based on reducing the particle size of the fat emulsion containing vitamin D to within 200-300 nm to obtain a fat emulsion vitamin D solution.

[0054] The above-prepared fat emulsion vitamin D liquid was filtered through 1.0um and 0.4um filter membranes respectively by high pressure, and lactose prepared in advance and filtered through 0.22um filter membrane was added, and freeze-dried. The obtained dry powder was crushed and sieved to obtain the vitamin D dry powder raw material wrapped in nano fat emulsion.

[0055] After passing the inspection, it is filled with nitrogen, packed in double-layer aluminum foil bags, and stored in the freezer.

[0056] Embodiment 2:

[0057] A vitamin D dry powder raw material encapsulated by nano fat emulsion, which is different from Example 1 in that the preparation process of colostrum comprises the following steps:

[0058] The same amount of soybean oil, vitamin D3, soybean lecithin, oleic acid, vitamin E and other raw materials and auxiliary materials as in Example 1 were dissolved in a mixed solvent of chloroform and methanol, and heated and stirred to completely dissolve the raw materials and auxiliary materials. The organic solvent was removed by a rotary evaporator to form a uniform phospholipid vitamin D film on the inner surface of the reactor. Then 800-850 mL of deionized water (pH 9.0) was added, and under the protection of nitrogen, the phospholipid film was hydrated and dispersed by heating and stirring to obtain colostrum.

[0059] Embodiment 3:

[0060] A vitamin D dry powder raw material encapsulated by nano fat emulsion, which is different from Example 1 in that the preparation process of colostrum comprises the following steps:

[0061] Under nitrogen protection, the same amount of soybean oil, vitamin D3, soybean lecithin, oleic acid, vitamin E and other raw materials and auxiliary materials as in Example 1 were added to anhydrous ethanol (about 10% of the final volume), and heated and stirred to completely dissolve the raw materials and auxiliary materials; the ethanol solution containing the raw materials and auxiliary materials was slowly added to 800-850 ml of deionized water (pH 9.0) and the above-mentioned amount of EDTA stirred at high speed by a shearing machine, the stirring speed was 3000 r / min, and stirring was continued for 25 minutes under nitrogen protection to obtain colostrum.

[0062] Example Part II: Different Drying Methods

[0063] Embodiment 4:

[0064] A vitamin D dry powder raw material wrapped in nano fat emulsion is different from Example 3 in that: the fat emulsion vitamin D liquid with excipients is filtered and then transported to a spray drying tower by a high-pressure pump for spray drying, the air inlet temperature is controlled at 100°C and the air outlet temperature is controlled at 70-90°C, and after drying, the nano fat emulsion wrapped in vitamin D dry powder raw material is obtained by crushing and sieving.

[0065] Example Part 3: Addition of coating stabilizer

[0066] Embodiment 5:

[0067] A vitamin D dry powder raw material wrapped in a nano fat emulsion is different from Example 3 in that: a coating stabilizer whose weight is 3% of the total weight of the prepared material is added to the prepared colostrum and then high-pressure homogenization is performed; the preparation method of the coating stabilizer comprises the following steps: adding carbon triglyceride and sturgeon skin gelatin to 50ML deionized water, heating to 50-60°C, stirring and dissolving, then adding a hydroxyapatite water suspension with a concentration of 0.3g / L under high-speed stirring, continuing to heat to 75°C, keeping the temperature for reaction for 2h, cooling to 25°C, continuing to stir for 2h, adding citric acid to adjust the pH to neutral, and obtaining the coating stabilizer; wherein the mass ratio of carbon triglyceride, sturgeon skin gelatin and hydroxyapatite is 1:0.5:0.06.

[0068] Embodiment 6:

[0069] A vitamin D dry powder raw material coated with nano fat emulsion, which is different from Example 5 in that the weight of the coating stabilizer is 2% of the total weight of the preparation material.

[0070] Embodiment 7:

[0071] A vitamin D dry powder raw material coated with nano fat emulsion, which is different from Example 5 in that the weight of the coating stabilizer is 4% of the total weight of the preparation material.

[0072] Embodiment 8:

[0073] A vitamin D dry powder raw material wrapped in nano fat emulsion is different from Example 5 in that the mass ratio of carbon triglyceride, sturgeon skin gelatin and hydroxyapatite is 1:0.4:0.04.

[0074] Embodiment 9:

[0075] A vitamin D dry powder raw material wrapped in nano fat emulsion is different from Example 5 in that the mass ratio of carbon triglyceride, sturgeon skin gelatin and hydroxyapatite is 1:0.6:0.08.

[0076] Example Part 4: Different Encapsulation Amounts of Vitamin D

[0077] Embodiment 10:

[0078] A vitamin D dry powder raw material wrapped in nano fat emulsion, which is different from Example 3 in that the mass percentage of vitamin D in the prepared material is 2%.

[0079] Embodiment 11:

[0080] A vitamin D dry powder raw material wrapped in nano fat emulsion, which is different from Example 3 in that the mass percentage of vitamin D in the prepared material is 10%.

[0081] Example Part V: Different Lecithin Packaging Materials

[0082] Embodiment 12:

[0083] A vitamin D dry powder raw material encapsulated by nano fat emulsion, which is different from Example 3 in that the lecithin encapsulating material is egg yolk lecithin.

[0084] Embodiment 13:

[0085] A vitamin D dry powder raw material encapsulated by nano fat emulsion, which is different from Example 3 in that the lecithin encapsulating material is hydrogenated soybean lecithin.

[0086] Embodiment 14:

[0087] A vitamin D dry powder raw material encapsulated by nano fat emulsion, which is different from Example 3 in that the lecithin packaging material is artificially synthesized lecithin, specifically the phospholipid dipalmitoylglycerol phosphatidylcholine.

[0088] Embodiment 15:

[0089] A vitamin D dry powder raw material encapsulated by nano fat emulsion, which is different from Example 3 in that the lecithin encapsulating material is sphingomyelin.

[0090] Example Part VI: Different Excipients

[0091] Embodiment 16:

[0092] A vitamin D dry powder raw material encapsulated by nano fat emulsion is different from that in Example 3 in that the excipient is mannitol.

[0093] Embodiment 17:

[0094] A vitamin D dry powder raw material encapsulated by nano fat emulsion is different from that of Example 3 in that the excipient is sorbitol.

[0095] Performance Test:

[0096] 1) Stability test: The recovery rate of vitamin D3 in the samples of Examples 1-17 was accurately measured using high performance liquid chromatography; after one year of low temperature storage, the recovery rate of vitamin D3 in the samples was measured again, and the measurement results are shown in Table 1;

[0097] 2) Determination of insoluble particles: The solubility of Examples 1-17 was determined according to the insoluble particle inspection method in section 0903 of the Chinese Pharmacopoeia 2020 Edition.

[0098] Table 1:

[0099]

[0100]

[0101] Combining Examples 1-3 and Table 1, it can be seen that the colostrum preparation methods of Examples 1-3 are all different. It can be seen from Table 1 that the recovery rate of vitamin D3 of the vitamin D dry powder raw material wrapped by the nano fat emulsion obtained in Examples 1-3 is all above 99%; and after being stored for one year, the recovery rate of vitamin D3 is still all above 95%. It can be obtained from the above that vitamin D3 is only slightly degraded in the process of preparing the vitamin D dry powder raw material wrapped by the nano fat emulsion, that is, it has good preparation stability and good long-term storage stability. And the insoluble particles of 10 μm or more in each 100 ml of Examples 1-3 are all below 5, and there are no insoluble particles of 25 μm or more, which shows that the vitamin D dry powder raw materials obtained in Examples 1-3 have good solubility, that is, they have good bioavailability.

[0102] Combining Examples 3-4 and Table 1, it can be seen that the drying method of Example 4 is different from that of Example 3. It can be seen from Table 1 that the recovery rate of vitamin D3 of the vitamin D dry powder raw material wrapped by the nano fat emulsion prepared in Example 4 is above 99%, and after one year of storage, the recovery rate is still above 95%; and the number of insoluble particles above 10 μm in each 100 ml is ≤5, and there are no insoluble particles above 25 μm. It can be seen from the above that the vitamin D dry powder raw materials prepared by freeze drying and spray drying have good preparation stability and long-term stability, and also have good solubility and bioavailability.

[0103] Combining Example 3, Example 5 and Table 1, it can be seen that a coating stabilizer is added to Example 5 before high-pressure homogenization. From Table 1, it can be seen that the recovery rate of vitamin D3 of the vitamin D dry powder raw material encapsulated by the nano fat emulsion prepared in Example 5 and the recovery rate after one year of storage are still higher than those in Example 3. It can be seen from the above that the addition of a coating stabilizer can further improve the preparation stability and long-term storage stability of the vitamin D dry powder raw material.

[0104] In combination with Example 5 and Examples 6-9, the weight ratio of the coating stabilizer used in Example 6-7 to the total weight of the prepared material is different from that in Example 5, and the mass ratios of carbon triglyceride, sturgeon skin gelatin and hydroxyapatite in Examples 8-9 are different from those in Example 5. As can be seen from Table 1, the vitamin D3 recovery rate of the vitamin D dry powder raw materials prepared in Examples 6-9 is above 99%, and after one year of storage, the recovery rate is still above 95%; and in each 100 ml of Examples 6-9, the insoluble particles above 10 μm are ≤5, and there are no insoluble particles above 25 μm.

[0105] Combining Example 3, Example 10-11 and Table 1, it can be seen that the mass percentage of vitamin D in the prepared material of Example 10-11 is different from that of Example 3. It can be seen from Table 1 that the vitamin D3 recovery rate of the vitamin D dry powder raw material prepared in Example 10-11 is above 99%, and after one year of storage, the recovery rate is still above 95%; and each 100 ml of Example 10-11 contains ≤5 insoluble particles above 10 μm, and there are no insoluble particles above 25 μm. It can be obtained from the above that the vitamin D dry powder raw material prepared by the vitamin D embedding amount within the scope of this application has good preparation stability and long-term stability, and also has good solubility and bioavailability.

[0106] Combining Example 3, Example 12-15 and Table 1, it can be seen that the lecithin used in Example 12-15 is different from that in Example 3. It can be seen from Table 1 that the vitamin D3 recovery rate of the vitamin D dry powder raw material prepared in Example 12-15 is above 99%, and after one year of storage, the recovery rate of vitamin D3 is still above 95%; and the insoluble particles of 10 μm or more in each 100 ml of Example 12-15 are ≤5, and there are no insoluble particles of 25 μm or more. It can be obtained from the above that the vitamin D dry powder raw material prepared by the lecithin packaging material in this application has good preparation stability and long-term stability, and also has good solubility and bioavailability.

[0107] Combining Example 3, Examples 16-17 and Table 1, it can be seen that the excipients used in Examples 16-17 are different from those in Example 3. It can be seen from Table 1 that the vitamin D3 recovery rate of the vitamin D dry powder raw materials prepared in Examples 16-17 is above 99%, and after one year of storage, the recovery rate is still above 95%; and the insoluble particles above 10 μm in each 100 ml of Examples 16-17 are ≤5 particles, and there are no insoluble particles above 25 μm. It can be seen from the above that the vitamin D dry powder raw materials prepared by the excipients of the present application have good preparation stability and long-term stability, and also have good solubility and bioavailability.

[0108] Example Part VII: Application of the Vitamin D Dry Powder Raw Material Encapsulated in Nanofat Emulsion to the Preparation of Oral Solid Preparations and Oral Liquid Preparations

[0109] Embodiment 18:

[0110] A vitamin D dry powder raw material wrapped in nano fat emulsion, magnesium stearate, the vitamin D dry powder raw material wrapped in nano fat emulsion prepared in Example 3, corn starch, pregelatinized starch, and calcium carbonate are added to a mixer in sequence according to the prescription amount in Table 2, and the material is discharged after mixing. The mixed material is sent to a dry granulator for granulation to obtain mixed particles; the vitamin D3 content is determined; and the loading amount is calculated according to the vitamin D3 content. The loading range is adjusted according to the vitamin D3 content determination result, and the mixed powder is filled into cellulose acetate phthalate enteric capsules to prepare capsules. After the finished product is tested and qualified, it is boxed, labeled, sealed, and stored.

[0111] Table 2:

[0112]

[0113] Embodiment 19:

[0114] A raw material of vitamin D dry powder wrapped in nano fat emulsion, carboxymethyl cellulose and glucose are dissolved in distilled water according to the prescription amount in Table 3, heated to 90°C, continuously heated for 30 minutes, and then cooled to 50°C, and then the fat emulsion D3 dry powder prepared in Example 3 is added, mixed evenly, and flavoring is added to prepare an oral solution preparation. After the finished product is tested and qualified, it is filled, labeled, sealed, and stored.

[0115] Table 3:

[0116] Ingredient name Prescription amount Fat emulsion D3 dry powder 400IU / 10ml Carboxymethyl cellulose 100L glucose 500g Food flavors Moderate

[0117] Performance Test:

[0118] The vitamin D3 recovery rate in the samples of Examples 18-19 was accurately determined using high performance liquid chromatography; the samples were stored at low temperature for 1 year, and samples were taken at 1, 2, 4, 6, 8, 10, and 12 months and the vitamin D3 recovery rate in the samples was determined again. The measurement results are shown in Table 4.

[0119] Table 4:

[0120]

[0121] Combining Examples 18-19 and Table 4, it can be seen that the recovery rate of vitamin D3 in the products of Examples 18-19 is maintained at more than 99%, and when the samples are stored at low temperature for 1 year, the vitamin D3 is maintained at more than 95%, that is, it is still within the qualified range. As can be seen from the above, the vitamin D dry powder raw material wrapped in the nano fat emulsion has no significant degradation in the capsule preparation process and the preparation process of the oral liquid preparation, that is, it is guaranteed that the vitamin D dry powder raw material of the present application can produce qualified solid preparations and oral liquid preparations, and the content of vitamin D3 meets the requirements of drug specifications during the one-year period of investigation.

[0122] Example Part 8: Application of Vitamin D Dry Powder Raw Material Encapsulated in Nanofat Emulsion in the Preparation of Injection

[0123] Embodiment 20:

[0124] A kind of vitamin D dry powder raw material wrapped by nano fat emulsion, the whole production process is carried out under nitrogen protection, the fat emulsion vitamin D3 dry powder prepared in Example 3 is added to 800-850ml injection water (pH8.0-8.5, water temperature 50°C) stirred at high speed (3000r / min) by a shearing machine according to the prescription amount of 1mL:7.5mg (300,000 IU) specification injection, and continued to stir for 10-30min under the condition of nitrogen protection, that is, vitamin D fat emulsion compound liquid is obtained. After homogenization by a high-pressure homogenizer 1-3 times, the pressure is 100MPa, and nitrogen filling is obtained to obtain a uniform fat emulsion vitamin D solution with a particle size range of 200-300nm, that is, an injection is prepared. After high-temperature rotation sterilization, lamp inspection, labeling, refrigerated storage.

[0125] Performance Testing:

[0126] The recovery rate of vitamin D3 in the sample of Example 20 was accurately determined using high performance liquid chromatography; the sample was stored at low temperature for 1 year, and samples were taken at 1, 2, 4, 6, 8, 10, and 12 months and the recovery rate of vitamin D3 and the particle size of the injection solution in the sample were determined again. The results are shown in Table 5.

[0127] Table 5:

[0128]

[0129]

[0130] Combining Example 20 and Table 5, it can be seen that the recovery rate of vitamin D3 in the product of Example 20 is above 99%, and when the sample is stored at low temperature for 1 year, vitamin D3 remains above 95%, that is, it is still within the qualified range. As can be seen from the above, the vitamin D dry powder raw material wrapped in the nano fat emulsion has no significant degradation during the preparation process of the injection, that is, it is guaranteed that the vitamin D dry powder raw material of the present application can produce qualified solid preparations and oral liquid preparations, and the content of vitamin D3 meets the requirements of drug specifications during the one-year period of investigation, and the particle size of the injection also meets the standard.

[0131] Example Part 9: Stability of Vitamin D Dry Powder Raw Material Encapsulated by Nanofat Emulsion

[0132] Comparative Example 1:

[0133] A method for preparing a vitamin D raw material comprises directly dissolving vitamin D3 in soybean oil to prepare a D3 oil solution.

[0134] Comparative Example 2:

[0135] A method for preparing a vitamin D raw material comprises directly dissolving vitamin D3 in an ethanol solution to prepare a D3 ethanol solution.

[0136] Comparative Example 3:

[0137] The difference from Example 3 is: a commercially available vitamin D3 solid dry powder sample.

[0138] Comparative Example 4:

[0139] The difference from Example 3 is that the prepared fat emulsion vitamin D liquid is not added with excipients and is not dried, and the vitamin D solution sample encapsulated by the nano fat emulsion is directly obtained.

[0140] Comparative Example 5:

[0141] The difference from Example 3 is that an equal amount of vitamin E in the preparation material is replaced by excipients.

[0142] Comparative Example 6:

[0143] The difference from Example 3 is that an equal amount of EDTA in the preparation material is replaced by excipients.

[0144] Performance testing:

[0145] 1) Using high performance liquid chromatography, the recovery rate of vitamin D3 in the samples of Example 3 and Comparative Examples 1-2 was accurately determined; the three samples were then placed in a light-proof oven maintained at 40° C., and samples were taken on the 5th and 10th days, and the recovery rate of vitamin D3 in the samples was determined again. The results are as follows: Figure 1 shown.

[0146] 2) Using high performance liquid chromatography to accurately measure the recovery rate of vitamin D3 in the samples of Example 3 and Comparative Example 1; then placing the samples of Example 3 and Comparative Example 1 in a dark, 60° C. oven, taking samples on the 20th, 40th, and 80th days, and measuring the recovery rate of vitamin D3 in the samples again, the measurement results are as follows: Figure 2 shown.

[0147] 3) The samples of Example 3 and Comparative Examples 1-6 were placed in a light-proof oven at different temperatures. Samples were taken after 5 days, and the reduction rate of the vitamin D3 content in each sample was measured and calculated. The calculation results are shown in Table 6.

[0148] Table 6:

[0149]

[0150] refer to Figure 1 It can be seen that the vitamin D dry powder raw material of Example 3 and the D3 oil solution of Comparative Example 1 can better maintain the preparation stability and short-term stability of vitamin D3 than the D3 ethanol solution of Comparative Example 2. On the 10th day, the degradation degree of vitamin D3 of the D3 oil solution of Comparative Example 1 and the D3 ethanol solution of Comparative Example 2 was much greater than that of the vitamin D dry powder raw material of Example 3, which shows that the fat emulsion encapsulation of the present application can better isolate vitamin D3 from the outside world, and has better protection and stability effects on vitamin D3.

[0151] Refer to Figure 2 It can be seen that although the vitamin D3 content in the sample of Example 3 of the present application is reduced, it still meets the quality standard requirements of health products during the test period; while the vitamin D3 content of the D3 oil solution sample of Comparative Example 1 showed a significant downward trend during the test period, and its vitamin D3 content no longer met the quality standard requirements of health products after 60 days.

[0152] In combination with Example 3, Comparative Examples 1-3 and Table 6, it can be seen that under the conditions of 40°C, protected from light and for 5 days, the vitamin D3 reduction rates of Comparative Examples 1-3 are much higher than those of Example 3, among which the D3 reduction rate of Comparative Example 2 is the highest, which shows that compared with D3 ethanol solution, D3 oil solution and commercially available vitamin D3 solid samples, the vitamin D dry powder raw material encapsulated by the nano-fat emulsion prepared in the present application can significantly improve the stability of vitamin D.

[0153] Combining Example 3, Comparative Examples 4-6 and Table 6, it can be seen that under the conditions of 40°C, protected from light, and for 5 days, the vitamin D3 reduction rates of Comparative Examples 4-6 are all higher than those of Example 3, indicating that when preparing the vitamin D dry powder raw material, adding excipients and drying, as well as antioxidants and ion chelating agents can effectively improve the stability of vitamin D.

[0154] It can also be seen from Table 6 that under the conditions of 60°C, dark, and 5 days, the vitamin D3 content of Example 3 only decreased by 2%, which is significantly less than the reduction rate of vitamin D3 of Comparative Examples 1-3 under the conditions of 40°C, dark, and 5 days. This shows that the vitamin D dry powder raw material encapsulated by the nano fat emulsion of the present application can improve the stability of the vitamin and ensure its stability during the pretreatment of the vitamin D raw material.

[0155] The above are all preferred embodiments of the present application, and the protection scope of the present application is not limited thereto. Therefore, all equivalent changes made based on the products, methods, and principles of the present application should be included in the protection scope of the present application.

Claims

1. A method for preparing a vitamin D dry powder raw material wrapped in a nano fat emulsion, characterized in that: The following steps are involved: Weigh the preparation material, which includes the following components and their mass percentages: 5% fat-soluble vitamin D3, 2% solubilizing component, 3% lecithin packaging material, 0.5% auxiliary packaging material, 0.03% antioxidant and 0.03% ion complexing agent, and the balance is excipient; The dissolution-aiding component is soybean oil, the lecithin packaging material is soybean lecithin, the auxiliary packaging material is oleic acid, the antioxidant is vitamin E, and the excipient is lactose; The solubilizing component, fat-soluble vitamin D3, lecithin packaging material, auxiliary packaging material, antioxidant and ion complexing agent are mixed to prepare colostrum; the colostrum is homogenized by high pressure until the particle size is less than 500 nm to obtain fat emulsion vitamin D liquid; Adding excipients to the fat emulsion vitamin D liquid, stirring and dissolving, filtering through a filter membrane, freeze-drying, and then crushing and sieving to obtain a nano fat emulsion-encapsulated vitamin D dry powder raw material; Adding 2-4% of the weight of a coating stabilizer to the colostrum and then homogenizing under high pressure, wherein the coating stabilizer is a gel prepared from triglyceride, sturgeon skin gelatin and hydroxyapatite in a mass ratio of 1: (0.4-0.6): (0.04-0.08); The preparation method of the coating stabilizer comprises the following steps: The carbon triglyceride and sturgeon skin gelatin are added to deionized water, heated to 50-60° C., stirred and dissolved, and then a 0.3 g / L hydroxyapatite aqueous suspension is added under high-speed stirring, and the temperature is continued to be raised to 70-80° C., kept for reaction for 1-2 hours, then cooled to 25° C., stirred for 1-2 hours, and citric acid is added to adjust the pH to neutral to obtain a coating stabilizer.

2. The method for preparing a vitamin D dry powder raw material wrapped by a nano fat emulsion according to claim 1, characterized in that: The particle size of the fat emulsion in the fat emulsion vitamin D medicinal solution is 50-300nm.

3. The method for preparing a vitamin D dry powder raw material wrapped by a nano fat emulsion according to claim 1, characterized in that: The homogenization pressure of the high-pressure homogenization is 95-110 MPa, and the homogenization times are 6-8 times.

4. A vitamin D dry powder raw material wrapped by nano fat emulsion, characterized in that: The vitamin D dry powder is prepared by the method for preparing the nano fat emulsion-encapsulated vitamin D dry powder raw material according to any one of claims 1 to 3.

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