High-stability dihydromyricetin nano-emulsion and preparation method thereof

Abstract

The invention belongs to the field of biological agents, and particularly relates to a high-stability dihydromyricetin nano-emulsion and a preparation method thereof. The dihydromyricetin nano-emulsion with high stability provided by the invention is prepared from the following components in parts by weight: 1 to 15.0 parts of dihydromyricetin, 1 to 3.0 parts of glyceride, 1 to 15 parts of an emulsifying agent, 0.05 to 0.1 part of a bacteriostatic agent, 1 to 3 parts of glycerol and 63.90 to 95.95 parts of water. According to the dihydromyricetin nano-emulsion and the preparation method thereof, a relatively strong emulsion interfacial film is formed through the compounding and synergistic effect of the surfactants from natural sources, the solubility of dihydromyricetin is greatly improved, the high-stability nano-emulsion with the average particle size within 200nm is obtained, and the defects in the prior art are overcome. The emulsion is stable for a long time and is nanoscale, the permeability of dihydromyricetin can be obviously improved, and the bioavailability is improved.

Description

TECHNICAL FIELD

[0001] The application belongs to the field of biological agents, and particularly relates to a high-stability dihydromyricetin nanoemulsion and a preparation method thereof. BACKGROUND

[0002] Dihydromyricetin (DHM) is a natural dihydroflavonol compound, which has good antioxidant, anticancer, anti-inflammatory, antimicrobial, liver protection, lipid and glucose metabolism regulation, and other pharmacological activities, and has high development value. However, DHM has low solubility and low permeability, resulting in low bioavailability, poor drug development, poor molecular stability, short half-life and other defects, which limit its wide application in clinical aspects.

[0003] In order to improve the above problems of DHM, in recent years, various new preparation technologies and new dosage forms have been used, such as forming liposomes, microemulsions, microcapsules, nano-micelles and the like, so as to significantly improve the stability, solubility and bioavailability of DHM.

[0004] Nanoemulsion is a kinetically stable, translucent or turbid liquid dispersion system, with a particle size of usually 20-300 nm, which can significantly improve the solubility and bioavailability of poorly soluble drugs. By loading dihydromyricetin in nanoemulsion, the water solubility and stability of dihydromyricetin can be effectively improved, the cell uptake efficiency can be enhanced, the drug degradation can be delayed, the half-life can be improved, and the bioavailability can be improved, so as to expand the application prospect of DHM in clinic, and have significant scientific research significance and market value. SUMMARY

[0005] Therefore, the technical problem to be solved by the present application is to provide a high-stability dihydromyricetin nanoemulsion and a preparation method thereof. The nanoemulsion is compounded with natural surfactants to form a strong emulsification interface film, which stably wraps dihydromyricetin in the oil phase, greatly improves the solubility of dihydromyricetin, forms a small-particle high-stability nanoemulsion with an average particle size of 200 nm, significantly increases the room temperature stability, and has good long-term storage stability. The present application overcomes the defects of the prior art, and the emulsion particles are nano-sized, which significantly improves the permeability of dihydromyricetin and improves the bioavailability thereof.

[0006] To solve the above technical problems, the technical scheme of the present application is as follows:

[0007] In one aspect, the present application provides a high-stability nanoemulsion of dihydromyricetin, comprising the following components in parts by weight: dihydromyricetin 1-15.0 parts, glyceride 1-3.0 parts, emulsifier 1-15 parts, bacteriostatic agent 0.05-0.1 parts, glycerol 1-3 parts, and water 63.90-95.95 parts; the emulsifier comprises Tween 80, soybean lecithin, and polyglycerol fatty acid monoester.

[0008] Preferably, the emulsifier comprises Tween 80 0.5-9 parts, soybean lecithin 0.05-3 parts, and polyglycerol fatty acid monoester 0.1-3 parts in parts by weight.

[0009] Further preferably, the weight ratio of Tween 80 to soybean lecithin is (1.5-3):1; specifically, it can be 1.5:1, 2:1, or 3:1; further preferably, the weight ratio of Tween 80 to soybean lecithin is 3:1.

[0010] Preferably, the glyceride comprises any one of natural or synthetic medium-chain triglyceride fatty acid ester, coconut oil, olive oil, avocado oil, or soybean oil.

[0011] Preferably, the bacteriostatic agent is any one of sodium benzoate or potassium sorbate.

[0012] In another aspect, the present application provides a preparation method of the high-stability nanoemulsion of dihydromyricetin, comprising the following steps:

[0013] (1) mixing the emulsifier and water, and fully magnetically stirring under heating conditions to fully dissolve the emulsifier, to obtain solution A; (2) ultrasonically treating glycerol, glyceride, and dihydromyricetin, and uniformly mixing to obtain glycerol-glyceride-dihydromyricetin mixture B; (3) after solution A is cooled to room temperature, glycerol-glyceride-dihydromyricetin mixture B is added dropwise into solution A, and homogenized emulsification is performed to obtain emulsion C; (4) adding a bacteriostatic agent into emulsion C, and magnetically stirring for a certain period of time to obtain the high-stability nanoemulsion of dihydromyricetin.

[0014] Preferably, the heating temperature in step (1) is 50-80°C; further preferably, the temperature is 60°C.

[0015] Preferably, the stirring time in step (1) is 10-40 minutes; further preferably, the stirring time is 20 minutes.

[0016] Preferably, the ultrasonic treatment in step (2) is performed under the following conditions: 2 minutes each time, 1 minute interval each time, 3 times of repetition, ultrasonic power of 500 W, and ultrasonic amplitude of 80%.

[0017] Preferably, the homogenization condition in step (3) is that the shearing rotation speed is 6000-18000 rpm, and the homogenization time is 10-30 minutes; further preferably, the homogenization condition is that the shearing rotation speed is 10000 rpm, and the homogenization time is 15 minutes. Preferably, the magnetic stirring condition in step (4) is that the stirring rotation speed is 50-1200 rpm, and the stirring time is 5-20 minutes; further preferably, the magnetic stirring condition is that the stirring rotation speed is 600 rpm, and the stirring time is 8 minutes.

[0018] The nanoemulsion of the application is characterized by particle size and microstructure, and an oil-in-water nanoemulsion with a particle size of less than 200 nm is obtained.

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

[0020] (1) The application significantly improves the solubility of DHM, and can increase the solubility of DHM by at least 100 times, the particle size of the DHM emulsion is less than 200 nm, which greatly improves the permeability of DHM and promotes the bioavailability, so as to promote the development of clinical application of DHM in antioxidant, anticancer, anti-inflammatory, antimicrobial, liver protection and other pharmacological activities. (2) The preparation process of the application adopts magnetic stirring, ultrasonic treatment, homogenization and other isothermal processes, which is simple and controllable, low in energy consumption, and convenient for industrial large-scale production; (3) The nanoemulsion prepared by the application can be used as a bioactive substance delivery system, providing a new idea for the design of medicine and functional food, and has a wide market application prospect. DETAILED DESCRIPTION

[0022] The specific embodiments of the application will be further described below. It should be noted that the description of the embodiments is used to help understand the application, but does not constitute a limitation on the application. In addition, the technical features involved in each embodiment of the application described below can be combined with each other as long as they do not conflict with each other.

[0023] If not specifically indicated, the chemical reagents and raw materials used in the examples are all conventional commercially available reagents, which do not need special extraction, and the products of different manufacturers do not have a significant effect on the results.

[0024] The specific embodiments of the application are as follows:

[0025] The application provides a preparation method of a high-stability nanoemulsion of dihydro-myrictanol, comprising the following steps:

[0026] (1) mixing the emulsifier and water, and fully magnetically stirring under heating condition to make the emulsifier fully dissolved, to obtain solution A; (2) ultrasonic treating the glycerol, glyceride and dihydromyricetin, and uniformly mixing to obtain glycerol-glyceride-dihydromyricetin mixture B; (3) after solution A is cooled to room temperature, glycerol-glyceride-dihydromyricetin mixture B is added dropwise into solution A, and homogenization emulsification is performed to obtain emulsion C; (4) adding bacteriostatic agent into emulsion C, and magnetically stirring for a certain time to obtain high-stability dihydromyricetin nanoemulsion.

[0027] The heating temperature in step (1) is 50-80℃, and the stirring time is 10-40 minutes; the ultrasonic condition in step (2) is: 2 minutes each time, 1 minute interval each time, 3 times in total, the ultrasonic power is 500w, and the ultrasonic amplitude is 80%; the homogenization condition in step (3) is: the shearing rotation speed is 6000-18000rpm, and the homogenization time is 10-30 minutes; the magnetic stirring condition in step (4) is: the stirring rotation speed is 50-1200r / min, and the stirring time is 5-20 minutes.

[0028] Example 1

[0029] The embodiment provides a high-stability dihydromyricetin nanoemulsion, which comprises, according to weight parts, 3 parts of dihydromyricetin, 2 parts of saturated caprylic acid triglyceride, 1.5 parts of Tween 80, 1 part of soybean lecithin, 0.4 part of polyglycerol fatty acid monoester, 0.03 part of potassium sorbate, 2 parts of glycerol and 88.57 parts of water.

[0030] The preparation method is: (1) weighing the ingredients: weighing various materials according to the proportion and reserving; (2) mixing Tween 80, soybean lecithin, polyglycerol fatty acid monoester and water, magnetically stirring under 50℃ heating condition for 20 minutes, to make the emulsifier fully dissolved, to obtain solution A; (3) ultrasonic treating glycerol, saturated caprylic acid triglyceride and dihydromyricetin, 2 minutes each time, 1 minute interval each time, 3 times in total, the ultrasonic power is 500w, and the ultrasonic amplitude is 80%, uniformly mixing to obtain glycerol-glyceride-dihydromyricetin mixture B; (4) after solution A is cooled to room temperature, glycerol-glyceride-dihydromyricetin mixture B is added dropwise into solution A, homogenization is performed at a shearing rotation speed of 8000rpm for 10 minutes, to make emulsion C; (5) adding potassium sorbate into emulsion C, magnetically stirring at a rotation speed of 500r / min for 5 minutes, to obtain high-stability dihydromyricetin nanoemulsion.

[0031] Example 2

[0032] The embodiment provides a kind of high stability dihydromyricetin nanoemulsion, according to weight fraction, including dihydromyricetin 4 parts, saturated capric acid triglyceride 3 parts, tween 80 4 parts, soybean lecithin 0.06 parts, polyglycerol fatty acid monoester 1 part, sodium benzoate 0.05 parts, glycerol 3 parts and water 80.89 parts.

[0033] The preparation method is: (1) weighing ingredients: after weighing various materials according to the ratio, reserve; (2) mix tween 80, soybean lecithin and polyglycerol fatty acid monoester and water, under the condition of heating at 50 DEG C, magnetic stirring for 30 minutes, make emulsifier fully dissolved to obtain solution A; (3) glycerol, saturated capric acid triglyceride and dihydromyricetin are treated by ultrasonic, after each ultrasonic 2 minutes, stop 1 minute, repeat 3 times, the ultrasonic power is 500w, ultrasonic amplitude is 80%, uniformly mixed to obtain glycerol-glyceride-dihydromyricetin mixture B; (4) after solution A is reduced to room temperature, glycerol-glyceride-dihydromyricetin mixture B is added to solution A, and the shear rotation speed is 9000rpm Homogeneous 15 minutes, emulsification obtains emulsion C; (5) add sodium benzoate to emulsion C, after stirring for 5 minutes under the condition of magnetic stirring rotation speed 600r / min, obtain the high stability dihydromyricetin nanoemulsion.

[0034] Example 3

[0035] The embodiment provides a kind of high stability dihydromyricetin nanoemulsion, according to weight fraction, including dihydromyricetin 5 parts, olive oil 2 parts, tween 80 3 parts, soybean lecithin 1 part, polyglycerol fatty acid monoester 1 part, potassium sorbate 0.03 parts, glycerol 2 parts and water 82.97 parts.

[0036] The preparation method is: (1) weighing ingredients: after weighing various materials according to the ratio, reserve; (2) mix tween 80, soybean lecithin and polyglycerol fatty acid monoester and water, under the condition of heating at 60 DEG C, magnetic stirring for 25 minutes, make emulsifier fully dissolved to obtain solution A; (3) glycerol, olive oil and dihydromyricetin are treated by ultrasonic, after each ultrasonic 2 minutes, stop 1 minute, repeat 3 times, the ultrasonic power is 500w, ultrasonic amplitude is 80%, uniformly mixed to obtain glycerol-glyceride-dihydromyricetin mixture B; (4) after solution A is reduced to room temperature, glycerol-glyceride-dihydromyricetin mixture B is added to solution A, and the shear rotation speed is 8000rpm Homogeneous 20 minutes, emulsification obtains emulsion C; (5) add potassium sorbate to emulsion C, after stirring for 6 minutes under the condition of magnetic stirring rotation speed 400r / min, obtain the high stability dihydromyricetin nanoemulsion.

[0037] Example 4

[0038] The embodiment provides a high-stability nanometer emulsion of dihydromyricetin, which comprises, according to weight parts, dihydromyricetin 6 parts, coconut oil 3 parts, Tween 80 5 parts, soybean lecithin 1 part, polyglycerol fatty acid monoester 1 part, potassium sorbate 0.03 part, glycerol 3 parts and water 75.97 parts.

[0039] The preparation method is as follows: (1) weighing ingredients: weighing various materials according to the proportion and reserving; (2) mixing Tween 80, soybean lecithin, polyglycerol fatty acid monoester and water, magnetically stirring for 10 minutes under the condition of heating at 50 DEG C, so that the emulsifier is fully dissolved to obtain solution A; (3) ultrasonic treating glycerol, coconut oil and dihydromyricetin, stopping for 1 minute after ultrasonic treating for 2 minutes each time, repeating for 3 times, the ultrasonic power is 500w, the ultrasonic amplitude is 80%, uniformly mixing to obtain glycerol-glyceride-dihydromyricetin mixed solution B; (4) after solution A is reduced to room temperature, glycerol-glyceride-dihydromyricetin mixed solution B is added dropwise into solution A, homogenizing for 10 minutes at a shearing rotating speed of 10000rpm, emulsifying to obtain emulsion C; (5) adding potassium sorbate into emulsion C, stirring for 5 minutes under the condition of magnetic stirring at a rotating speed of 800r / min, so that the high-stability nanometer emulsion of dihydromyricetin is obtained.

[0040] Embodiment 5

[0041] The embodiment provides a high-stability nanometer emulsion of dihydromyricetin, which comprises, according to weight parts, dihydromyricetin 8 parts, soybean oil 3 parts, Tween 80 6 parts, soybean lecithin 1 part, polyglycerol fatty acid monoester 1 part, sodium benzoate 0.05 part, glycerol 3 parts and water 71.95 parts.

[0042] The preparation method is as follows: (1) weighing ingredients: weighing various materials according to the proportion and reserving; (2) mixing Tween 80, soybean lecithin, polyglycerol fatty acid monoester and water, magnetically stirring for 10 minutes under the condition of heating at 50 DEG C, so that the emulsifier is fully dissolved to obtain solution A; (3) ultrasonic treating glycerol, coconut oil and dihydromyricetin, stopping for 1 minute after ultrasonic treating for 2 minutes each time, repeating for 3 times, the ultrasonic power is 500w, the ultrasonic amplitude is 80%, uniformly mixing to obtain glycerol-glyceride-dihydromyricetin mixed solution B; (4) after solution A is reduced to room temperature, glycerol-glyceride-dihydromyricetin mixed solution B is added dropwise into solution A, homogenizing for 10 minutes at a shearing rotating speed of 10000rpm, emulsifying to obtain emulsion C; (5) adding potassium sorbate into emulsion C, stirring for 5 minutes under the condition of magnetic stirring at a rotating speed of 800r / min, so that the high-stability nanometer emulsion of dihydromyricetin is obtained.

[0043] Embodiment 6

[0044] This embodiment provides a highly stable dihydromyricetin nanoemulsion, which, by weight, comprises 12 parts dihydromyricetin, 3 parts coconut oil, 6 parts Tween 80, 2 parts soybean lecithin, 1 part polyglycerol fatty acid monoester, 0.05 parts potassium sorbate, 2 parts glycerol, and 67.95 parts water.

[0045] The preparation method is as follows: (1) Weighing and mixing: Weigh all kinds of materials according to the proportion and set aside; (2) Mix Tween 80, soybean lecithin and polyglycerol fatty acid monoester with water, and stir magnetically for 20 minutes under heating at 70°C to fully dissolve the emulsifier and obtain solution A; (3) Sonicate glycerol, coconut oil and dihydromyricetin, sonicate for 2 minutes each time and stop for 1 minute, repeat 3 times, the ultrasonic power is 500w and the ultrasonic amplitude is 80%, and after uniform mixing, obtain glycerol-glyceryl ester-dihydromyricetin mixture B; (4) After cooling the solution A to room temperature, add glycerol-glyceryl ester-dihydromyricetin mixture B dropwise to solution A, homogenize at a shear speed of 10000rpm for 10 minutes, and emulsify to obtain emulsion C; (5) Add potassium sorbate to emulsion C, stir at a magnetic stirring speed of 600 rpm for 6 minutes to obtain a nanoemulsion of highly stable dihydromyricetin.

Claims

1. A highly stable dihydromyricetin nanoemulsion, characterized in that, The product comprises, by weight, the following components: 1-15.0 parts of dihydromyricetin, 1-3.0 parts of glycerides, 1-15 parts of emulsifier, 0.05-0.1 parts of antibacterial agent, 1-3 parts of glycerin, and 63.90-95.95 parts of water.

2. The highly stable dihydromyricetin nanoemulsion according to claim 1, characterized in that, The emulsifier includes Tween 80, soybean lecithin, and polyglycerol fatty acid monoester; the emulsifier, by weight, includes 0.5-9 parts of Tween 80, 0.05-3 parts of soybean lecithin, and 0.1-3 parts of polyglycerol fatty acid monoester.

3. The highly stable dihydromyricetin nanoemulsion according to claim 1, characterized in that, The glycerides include any one of natural or synthetic medium-chain triglyceride fatty acid esters, coconut oil, olive oil, avocado oil, or soybean oil.

4. The highly stable dihydromyricetin nanoemulsion according to claim 1, characterized in that, The antibacterial agent is either sodium benzoate or potassium sorbate.

5. A method for preparing a highly stable dihydromyricetin nanoemulsion as described in claim 1, characterized in that, The preparation method includes the following steps: (1) mixing emulsifier and water, and stirring magnetically under heating conditions to fully dissolve the emulsifier to obtain solution A; (2) ultrasonically treating glycerol, glyceryl ester and dihydromyricetin, and mixing them uniformly to obtain glycerol-glyceryl ester-dihydromyricetin mixture B; (3) cooling solution A to room temperature, adding glycerol-glyceryl ester-dihydromyricetin mixture B dropwise to solution A, and homogenizing and emulsifying to obtain emulsion C; (4) adding antibacterial agent to emulsion C, and stirring magnetically for a certain time to obtain a nanoemulsion of highly stable dihydromyricetin.

6. The highly stable dihydromyricetin nanoemulsion according to claim 5, characterized in that, In step (1), the heating temperature is 50~180℃ and the stirring time is 10~40 minutes.

7. The highly stable dihydromyricetin nanoemulsion according to claim 5, characterized in that, The homogenization conditions in step (3) are: shearing speed of 6000~18000 rpm and homogenization time of 10~30 minutes.

8. The highly stable dihydromyricetin nanoemulsion according to claim 5, characterized in that, In step (4), the magnetic stirring speed is 50~1200 rpm and the stirring time is 5~20 minutes.