Hyaluronic acid-casein-vitamin D3 water-soluble particles as well as preparation method and application thereof

Abstract

The invention discloses hyaluronic acid-casein-vitamin D3 water-soluble particles and a preparation method and application thereof, and relates to the technical field of water-based preparations and vitamins, the water-soluble particles are obtained by including vitamin D3, casein and amphiphilic hyaluronic acid with sulfobutyl-beta-cyclodextrin; the amphiphilic hyaluronic acid is long-chain alkyl modified hyaluronic acid; long-chain alkyl is introduced into a hyaluronic acid structure, hyaluronic acid is endowed with excellent amphiphilic performance, casein and amphiphilic hyaluronic acid are combined for use, the stability of vitamin D3 is effectively improved, the bioavailability of vitamin D3 is also improved, and the utilization efficiency of the product is higher; a hydrophobic cavity in sulfobutyl-beta-cyclodextrin effectively comprises vitamin D3, an external hydrophilic structure can be combined with hydrophilic structures in casein and amphiphilic hyaluronic acid, a hydrophobic long chain in modified hyaluronic acid can be inserted into the hydrophobic cavity, and the wrapping effect is better.

Description

Technical Field

[0001] The present invention relates to the technical field of aqueous preparations and vitamins, and in particular to a hyaluronic acid-casein-vitamin D 3 Water-soluble particles and preparation method and application thereof. Background Art

[0002] Vitamin D 3 It is an important form of vitamin D, which can effectively promote calcium absorption and bone metabolism and promote bone mineralization. With the change of people's lifestyle, people's dietary structure and working environment have undergone tremendous changes, resulting in the widespread presence of vitamin D in the body. 3 Symptoms of deficiency. Vitamin D 3 Vitamin D deficiency can cause serious diseases, including osteoporosis in infants and young children, rickets in the elderly, etc. 3 It is also related to respiratory and cardiovascular diseases. 3 It is fat-soluble, insoluble in water, and easily soluble in most organic solvents. Its structure is unstable and is easily changed under light, acidic and high temperature conditions. At present, many researchers have prepared new types of vitamin D 3 Granules are used to improve their water solubility and stability, but the stability of existing granules still needs to be improved, the shelf life is short, and there is a small amount of precipitation after dissolving in water. In addition, some vitamin D 3 The particles are made into an oily oral spray by adding rice bran oil, but the greasy feeling of the oil droplets makes the mouth feel bad, which is not good for children and adolescents.

[0003] Therefore, it is urgent to develop a vitamin D that is highly water-soluble, stable, tastes good, and is easily absorbed by the human body. 3 Particles. Summary of the invention

[0004] In order to solve the above problems, the present invention aims to provide a hyaluronic acid-casein-vitamin D 3 Water-soluble particles and preparation method and application thereof.

[0005] In order to achieve the above object, the present invention is implemented through the following technical solutions:

[0006] A hyaluronic acid-casein-vitamin D 3 Water-soluble granules containing vitamin D encapsulated by sulfobutyl-β-cyclodextrin 3 , casein and amphiphilic hyaluronic acid; the amphiphilic hyaluronic acid is long-chain alkyl-modified hyaluronic acid.

[0007] A hyaluronic acid-casein-vitamin D 3 The method for preparing water-soluble particles comprises the following steps:

[0008] ① Preparation of amphiphilic hyaluronic acid: by weight, weigh 18 to 22 parts of hyaluronic acid and 2.5 to 4.5 parts of long-chain alkylamine, add them to 1000 to 2000 parts of organic solvent, add 3.2 to 5.6 parts of condensing agent and 0.02 to 0.06 parts of 4-dimethylaminopyridine under stirring, react at 10 to 40° C. for 10 to 18 hours, filter, pour the filtrate into 2000 to 4000 parts of alcohol solvent, stir for 20 to 40 minutes, filter, wash the obtained solid with 300 to 500 parts of alcohol solvent, and dry to obtain amphiphilic hyaluronic acid;

[0009] ② Add 15 to 25 parts of the amphiphilic hyaluronic acid obtained in step ①, 38 to 46 parts of casein, and 40 to 50 parts of sulfobutyl-β-cyclodextrin to 10,000 to 12,000 parts of deionized water to obtain an aqueous phase for later use;

[0010] Take 5-10 servings of vitamin D 3 Add to 120-250 parts of medium chain triglycerides to obtain an oil phase for later use;

[0011] The obtained aqueous phase and oil phase are mixed, homogenized at a pressure of 400-600 psi and a temperature of 30-45° C. for 30-40 minutes to obtain an emulsion, and the obtained emulsion is spray-dried to obtain hyaluronic acid-casein-vitamin D 3 Water-soluble granules.

[0012] Preferably, the long-chain alkylamine is 1-aminododecane or 1-aminotetradecane.

[0013] Preferably, the organic solvent is dimethyl sulfoxide or N,N-dimethylformamide.

[0014] Preferably, the condensing agent is 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride or N,N-dicyclohexylcarbodiimide.

[0015] Preferably, the alcohol solvent is methanol or ethanol.

[0016] Preferably, the homogenization pressure is 500-600 psi and the temperature is 35-40°C.

[0017] Preferably, the spray drying air inlet temperature is 70°C to 95°C, the atomization pressure is 0.15MPa to 0.25MPa, and the air outlet temperature is 65°C to 80°C.

[0018] The present invention also includes a hyaluronic acid-casein-vitamin D 3 Application of water-soluble particles as vitamin D 3 Supplements.

[0019] Compared with the prior art, the present invention has the following advantages:

[0020] Hyaluronic acid-casein-vitamin D of the present invention 3 Water-soluble particles introduce long-chain alkyl groups into the hyaluronic acid structure, giving hyaluronic acid excellent amphiphilic properties. The combined use of casein and amphiphilic hyaluronic acid effectively improves vitamin D 3 While improving the stability, it also improves its bioavailability, and the utilization efficiency of the product is higher. Sulfonbutyl-β-cyclodextrin is used as an effective additive. The hydrophobic cavity inside sulfonbutyl-β-cyclodextrin effectively includes vitamin D 3 The external hydrophilic structure can be combined with the hydrophilic structure in casein and amphiphilic hyaluronic acid, while the hydrophobic long chain in the modified hyaluronic acid can be inserted into the hydrophobic cavity, which has a better wrapping effect, good taste, strong palatability, and is easy for the human body to absorb. DETAILED DESCRIPTION

[0021] The purpose of the present invention is to provide a hyaluronic acid-casein-vitamin D 3 The water-soluble particles, preparation method and application thereof are further described below in conjunction with specific embodiments.

[0022] Example 1

[0023] A hyaluronic acid-casein-vitamin D 3 The method for preparing water-soluble particles comprises the following steps:

[0024] ① Preparation of amphiphilic hyaluronic acid: weigh 18 g of hyaluronic acid and 2.5 g of 1-aminododecane, add them to 1000 g of dimethyl sulfoxide, add 3.2 g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.02 g of 4-dimethylaminopyridine under stirring, react at 10° C. for 18 hours, filter, pour the filtrate into 2000 g of ethanol, stir for 20 minutes, filter, wash the obtained solid with 300 g of ethanol, and dry to obtain amphiphilic hyaluronic acid;

[0025] ② Add 15 g of the amphiphilic hyaluronic acid obtained in step ①, 38 g of casein, and 40 g of sulfobutyl-β-cyclodextrin to 10,000 g of deionized water to obtain an aqueous phase for later use;

[0026] 5g Vitamin D 3 Add to 120 g of medium chain triglyceride to obtain an oil phase, which is set aside;

[0027] The obtained aqueous phase and oil phase were mixed, homogenized at a pressure of 400 psi and a temperature of 30° C. for 40 minutes to obtain an emulsion, and the obtained emulsion was spray-dried to obtain hyaluronic acid-casein-vitamin D 3Water-soluble particles; the spray drying air inlet temperature is 70°C, the atomization pressure is 0.25MPa, and the air outlet temperature is 65°C.

[0028] Example 2

[0029] A hyaluronic acid-casein-vitamin D 3 The method for preparing water-soluble particles comprises the following steps:

[0030] ① Preparation of amphiphilic hyaluronic acid: weigh 22 g of hyaluronic acid and 4.5 g of 1-aminotetradecane, add them to 2000 g of N,N-dimethylformamide, add 5.6 g of N,N-dicyclohexylcarbodiimide and 0.06 g of 4-dimethylaminopyridine under stirring, react at 40° C. for 10 hours, filter, pour the filtrate into 4000 g of methanol, stir for 40 minutes, filter, wash the obtained solid with 500 g of methanol, and dry to obtain amphiphilic hyaluronic acid;

[0031] ② Add 25 g of the amphiphilic hyaluronic acid obtained in step ①, 46 g of casein, and 50 g of sulfobutyl-β-cyclodextrin to 12000 g of deionized water to obtain an aqueous phase for later use;

[0032] 10g vitamin D 3 Add to 250 g of medium chain triglyceride to obtain an oil phase, which is set aside;

[0033] The obtained aqueous phase and oil phase were mixed, homogenized at a pressure of 600 psi and a temperature of 45° C. for 30 minutes to obtain an emulsion, and the obtained emulsion was spray-dried to obtain hyaluronic acid-casein-vitamin D 3 Water-soluble particles; the spray drying air inlet temperature is 95°C, the atomization pressure is 0.15MPa, and the air outlet temperature is 80°C.

[0034] Example 3

[0035] A hyaluronic acid-casein-vitamin D 3 The method for preparing water-soluble particles comprises the following steps:

[0036] ① Preparation of amphiphilic hyaluronic acid: 19 g hyaluronic acid and 4 g 1-aminododecane were weighed and added to 1200 g dimethyl sulfoxide, and 3.6 g 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.03 g 4-dimethylaminopyridine were added under stirring, and the mixture was stirred at 20° C. for 12 hours, filtered, and the filtrate was poured into 2200 g methanol, stirred for 25 minutes, filtered, and the obtained solid was washed with 320 g methanol and dried to obtain amphiphilic hyaluronic acid;

[0037] ② Add 18 g of the amphiphilic hyaluronic acid obtained in step ①, 40 g of casein, and 42 g of sulfobutyl-β-cyclodextrin to 10500 g of deionized water to obtain an aqueous phase for later use;

[0038] 6g of vitamin D 3 Add to 150 g of medium chain triglyceride to obtain an oil phase, which is set aside;

[0039] The obtained aqueous phase and oil phase were mixed, homogenized at a pressure of 500 psi and a temperature of 35° C. for 32 minutes to obtain an emulsion, and the obtained emulsion was spray-dried to obtain hyaluronic acid-casein-vitamin D 3 Water-soluble particles; the spray drying air inlet temperature is 75°C, the atomization pressure is 0.18MPa, and the air outlet temperature is 70°C.

[0040] Example 4

[0041] A hyaluronic acid-casein-vitamin D 3 The method for preparing water-soluble particles comprises the following steps:

[0042] ① Preparation of amphiphilic hyaluronic acid: weigh 21 g of hyaluronic acid and 3.5 g of 1-aminotetradecane, add them to 1400 g of dimethyl sulfoxide, add 3.8 g of N,N-dicyclohexylcarbodiimide and 0.04 g of 4-dimethylaminopyridine under stirring, react at 35° C. for 16 hours, filter, pour the filtrate into 3800 g of ethanol, stir for 25 minutes, filter, wash the obtained solid with 340 g of ethanol, and dry to obtain amphiphilic hyaluronic acid;

[0043] ② Add 24 g of the amphiphilic hyaluronic acid obtained in step ①, 42 g of casein, and 48 g of sulfobutyl-β-cyclodextrin to 11000 g of deionized water to obtain an aqueous phase for later use;

[0044] 8g of vitamin D 3 Add to 220 g of medium chain triglyceride to obtain an oil phase, which is set aside;

[0045] The obtained aqueous phase and oil phase were mixed, homogenized at a pressure of 580 psi and a temperature of 40° C. for 38 minutes to obtain an emulsion, and the obtained emulsion was spray-dried to obtain hyaluronic acid-casein-vitamin D 3 Water-soluble particles; the spray drying air inlet temperature is 90°C, the atomization pressure is 0.22MPa, and the air outlet temperature is 75°C.

[0046] Example 5

[0047] A hyaluronic acid-casein-vitamin D 3 The method for preparing water-soluble particles comprises the following steps:

[0048] ① Preparation of amphiphilic hyaluronic acid: weigh 20 g of hyaluronic acid and 3 g of 1-aminododecane, add them to 1500 g of N,N-dimethylformamide, add 4 g of N,N-dicyclohexylcarbodiimide and 0.05 g of 4-dimethylaminopyridine under stirring, react at 35°C for 15 hours, filter, pour the filtrate into 3000 g of ethanol, stir for 30 minutes, filter, wash the obtained solid with 400 g of ethanol, and dry to obtain amphiphilic hyaluronic acid;

[0049] ② Add 20 g of the amphiphilic hyaluronic acid obtained in step ①, 40 g of casein, and 45 g of sulfobutyl-β-cyclodextrin to 11000 g of deionized water to obtain an aqueous phase for later use;

[0050] 7g of vitamin D 3 Add to 150 g of medium chain triglyceride to obtain an oil phase, which is set aside;

[0051] The obtained aqueous phase and oil phase were mixed, homogenized at a pressure of 550 psi and a temperature of 38° C. for 35 minutes to obtain an emulsion, and the obtained emulsion was spray-dried to obtain hyaluronic acid-casein-vitamin D 3 Water-soluble particles; the spray drying air inlet temperature is 80°C, the atomization pressure is 0.20MPa, and the air outlet temperature is 70°C.

[0052] Example 6

[0053] A hyaluronic acid-casein-vitamin D 3 The method for preparing water-soluble particles comprises the following steps:

[0054] ① Preparation of amphiphilic hyaluronic acid: weigh 20 g of hyaluronic acid and 4 g of 1-aminotetradecane, add them to 1500 g of N,N-dimethylformamide, add 5 g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.04 g of 4-dimethylaminopyridine under stirring, react at 30° C. for 14 hours, filter, pour the filtrate into 3500 g of methanol, stir for 30 minutes, filter, wash the obtained solid with 450 g of methanol, and dry to obtain amphiphilic hyaluronic acid;

[0055] ② Add 20 g of the amphiphilic hyaluronic acid obtained in step ①, 40 g of casein, and 45 g of sulfobutyl-β-cyclodextrin to 11500 g of deionized water to obtain an aqueous phase for later use;

[0056] 6g of vitamin D 3 Add to 200 g of medium chain triglyceride to obtain an oil phase, which is set aside;

[0057] The obtained aqueous phase and oil phase were mixed, homogenized at a pressure of 500 psi and a temperature of 40° C. for 35 minutes to obtain an emulsion, and the obtained emulsion was spray-dried to obtain hyaluronic acid-casein-vitamin D3 Water-soluble particles; the spray drying air inlet temperature is 90°C, the atomization pressure is 0.20MPa, and the air outlet temperature is 70°C.

[0058] 1. Particle size and water solubility test

[0059] The hyaluronic acid-casein-vitamin D obtained in Examples 1 to 6 of the present invention was measured using a Malvern particle size analyzer. 3 The particle size of the water-soluble particles is shown in Table 1. The particle size of the products prepared by the present invention is about 100 μm and is evenly distributed. 3 Add 1 g of water-soluble granules into 50 mL of water, stir to dissolve, and observe the condition of the solution.

[0060] Table 1 Particle size and water solubility test results

[0061] Particle size / μm Aqueous solution state Example 1 103±34μm Clarity and transparency Example 2 129±39μm Clarity and transparency Example 3 116±25μm Clarity and transparency Example 4 122±51μm Clarity and transparency Example 5 120±24μm Clarity and transparency Example 6 114±35μm Clarity and transparency

[0062] It can be seen from the results in Table 1 that the hyaluronic acid-casein-vitamin D obtained by the present invention 3 The water-soluble particles have a particle size of about 100 μm and are evenly distributed. After being dissolved in water, they are all clear and transparent without obvious precipitation and have good water solubility.

[0063] 2. Long-term stability test

[0064] Vitamin D 3 Content determination: Hyaluronic acid-casein-vitamin D 3 An appropriate amount of water-soluble particles was added to a centrifuge tube, and an appropriate amount of n-hexane was added. The tube was shaken for 10 minutes, and solid impurities were removed by filtering with a microporous membrane with a diameter of 0.45 μm. 100 mL of the filtrate was taken and vitamin D was determined on a high performance liquid chromatograph according to the method described in the imported standard (standard number JX20070088). 3 content.

[0065] Hyaluronic acid-casein-vitamin D 3 The water-soluble granules were placed at a temperature of 40°C and a relative humidity of 75% for 6 months. Vitamin D was measured at the end of the 3rd and 6th months. 3 The results are shown in Table 2. After 6 months, the hyaluronic acid-casein-vitamin D 3 Vitamin D in water-soluble granules 3 The contents of Example 2 have all decreased, but the contents are still greater than 97%, and the stability is good. Example 2 has the highest content and the best stability.

[0066] Table 2 Stability test results

[0067]

[0068] 3. Light stability test and thermal stability test

[0069] The hyaluronic acid-casein-vitamin D obtained in Examples 1 to 6 3 The water-soluble particles were divided into two parts. One part was subjected to the light and temperature test. The hyaluronic acid-casein-vitamin D obtained in Examples 1 to 6 without packaging was tested according to the influencing factors in the Guiding Principles for Stability Tests of APIs and Preparations in the Appendix of the 2020 Edition of the Chinese Pharmacopoeia. 3 The water-soluble granules were placed under light conditions for 10 days, and samples were taken on the 0th and 10th days to measure vitamin D 3 The results are shown in Table 3. The vitamin D prepared in the embodiment of the present invention 3 The particles all have good light stability. After being placed under light for 10 days, they still have a relatively high content, among which the sample prepared in Example 6 has the highest content.

[0070] Another batch was subjected to a thermal stability test, which was placed at 60°C for 10 days. Samples were taken on the 0th and 10th days to determine the vitamin D 3 The results are shown in Table 3. The hyaluronic acid-casein-vitamin D prepared in the embodiment of the present invention 3 The water-soluble particles all have good thermal stability and still have a relatively high content after being placed at 60° C. for 10 days, among which the sample prepared in Example 6 has the highest content.

[0071] Table 3 Results of photostability test and thermal stability test

[0072] <![CDATA[0-day VD 3 / %]]> <![CDATA[Light stability VD 3 / %]]> <![CDATA[Thermal stability VD 3 / %]]> Example 1 105.4 98.5 98.3 Example 2 106.4 99.2 99.1 Example 3 105.2 98.4 98.2 Example 4 105.8 98.7 98.5 Example 5 104.8 98.1 97.6 Example 6 105.3 98.4 98.3

Claims

1. A hyaluronic acid-casein-vitamin D3 water-soluble particle, characterized in that: The invention is obtained by including vitamin D3, casein and amphiphilic hyaluronic acid in sulfobutyl-β-cyclodextrin; the amphiphilic hyaluronic acid is a long-chain alkyl-modified hyaluronic acid.

2. A method for preparing hyaluronic acid-casein-vitamin D3 water-soluble particles, characterized in that: The following steps are involved: ① Preparation of amphiphilic hyaluronic acid: by weight, weigh 18 to 22 parts of hyaluronic acid and 2.5 to 4.5 parts of long-chain alkylamine, add them to 1000 to 2000 parts of organic solvent, add 3.2 to 5.6 parts of condensing agent and 0.02 to 0.06 parts of 4-dimethylaminopyridine under stirring, react at 10 to 40° C. for 10 to 18 hours, filter, pour the filtrate into 2000 to 4000 parts of alcohol solvent, stir for 20 to 40 minutes, filter, wash the obtained solid with 300 to 500 parts of alcohol solvent, and dry to obtain amphiphilic hyaluronic acid; ② Add 15 to 25 parts of the amphiphilic hyaluronic acid obtained in step ①, 38 to 46 parts of casein, and 40 to 50 parts of sulfobutyl-β-cyclodextrin to 10,000 to 12,000 parts of deionized water to obtain an aqueous phase for later use; Add 5-10 parts of vitamin D3 to 120-250 parts of medium chain triglycerides to obtain an oil phase for later use; The obtained aqueous phase and oil phase are mixed, and homogenized at a pressure of 400-600 psi and a temperature of 30-45° C. for 30-40 minutes to obtain an emulsion. The obtained emulsion is spray-dried to obtain hyaluronic acid-casein-vitamin D3 water-soluble particles.

3. The method for preparing a hyaluronic acid-casein-vitamin D3 water-soluble particle according to claim 2, characterized in that: The long-chain alkylamine is 1-aminododecane or 1-aminotetradecane.

4. The method for preparing a hyaluronic acid-casein-vitamin D3 water-soluble particle according to claim 2, characterized in that: The organic solvent is dimethyl sulfoxide or N,N-dimethylformamide.

5. The method for preparing a hyaluronic acid-casein-vitamin D3 water-soluble particle according to claim 2, characterized in that: The condensing agent is 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride or N,N-dicyclohexylcarbodiimide.

6. The method for preparing a hyaluronic acid-casein-vitamin D3 water-soluble particle according to claim 2, characterized in that: The alcohol solvent is methanol or ethanol.

7. The method for preparing a hyaluronic acid-casein-vitamin D3 water-soluble particle according to claim 2, characterized in that: The homogenization pressure is 500-600 psi and the temperature is 35-40°C.

8. The method for preparing a hyaluronic acid-casein-vitamin D3 water-soluble particle according to claim 2, characterized in that: The spray drying air inlet temperature is 70°C to 95°C, the atomization pressure is 0.15MPa to 0.25MPa, and the air outlet temperature is 65°C to 80°C.

9. The use of the hyaluronic acid-casein-vitamin D3 water-soluble particles according to claim 1, characterized in that: As a vitamin D3 supplement.

Images