Water-in-oil-in-water pickering emulsion loaded with arnica extract and method for its preparation

Abstract

The application discloses a water-in-oil-in-water Pickering emulsion loaded with Artemisia rupestris L. extract and a preparation method thereof, relates to the field of Pickering emulsion preparation, and is composed of a water solution of Artemisia rupestris L. natural active extract, lecithin, edible oil and sodium alginate-lysozyme protein amphiphilic composite nanoparticles. The application utilizes the characteristics of the Pickering emulsion, first, uses edible oil and lecithin to wrap the water solution of Artemisia rupestris L. natural active substance in a water-in-oil form, and then uses sodium alginate-lysozyme protein amphiphilic composite nanoparticles to wrap the water solution again, so as to form a water-in-oil-in-water Pickering emulsion, thereby protecting the Artemisia rupestris L. natural active extract, improving the stability and bioavailability of the Artemisia rupestris L. natural active substance, and making the prepared Pickering emulsion rich in Artemisia rupestris L. natural active substance have a more abundant nutritional value compared with traditional Pickering emulsions, and meet the diversified demand of the market for functional foods.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of Pickering emulsion preparation, in particular to a water-in-oil-in-water Pickering emulsion loaded with Artemisia selengensis extracts and a preparation method thereof. BACKGROUND

[0002] Pickering emulsion refers to an emulsion prepared by replacing traditional surfactants with solid particles as stabilizers. Compared with traditional emulsions stabilized by small molecule surfactants, Pickering emulsion has unique advantages: good emulsification effect, thereby saving cost; generally no post-treatment, environmentally friendly; strong emulsion stability, not easily affected by external factors such as temperature, long-term stability; therefore, the oil / water interface in Pickering emulsion also provides an ideal place for the self-assembly of solid particles, which can be used to prepare hollow structures, functional microcapsules and porous structures, etc.

[0003] Artemisia selengensis is rich in flavonoids, volatile oils, polysaccharides, alkaloids and other chemical components, and has the effects of antioxidant, antitumor, antibacterial and insecticidal, etc. However, functional active substances of Artemisia selengensis such as flavonoids and terpenes are easily oxidized, and there is currently a lack of research on protection technologies such as embedding of functional active substances of Artemisia selengensis, which makes it difficult for Artemisia selengensis deep processing products to be applied in the food or pharmaceutical industry. Therefore, it is necessary to provide a water-in-oil-in-water Pickering emulsion loaded with Artemisia selengensis extracts and a preparation method thereof to solve the above problems. SUMMARY

[0004] To solve the above technical problems, the present application provides the following technical solutions:

[0005] The water-in-oil-in-water Pickering emulsion loaded with Artemisia selengensis extracts is composed of an Artemisia selengensis natural active extract aqueous solution, lecithin, edible oil and sodium alginate-lysozyme protein amphiphilic composite nanoparticles.

[0006] The present application also discloses a preparation method of the water-in-oil-in-water Pickering emulsion loaded with Artemisia selengensis extracts, comprising the following steps:

[0007] Step one, after the Artemisia selengensis natural active extract aqueous solution, edible oil and lecithin are subjected to magnetic stirring for 30 min, an Artemisia selengensis natural active substance in the form of water-in-oil is prepared, wherein the volume ratio of the Artemisia selengensis natural active extract aqueous solution to the edible oil is 1:2, and the amount of lecithin is 3-5 wt% of the mass of the edible oil;

[0008] Step two, the water-in-oil form of natural active material of Artemisia selengensis and sodium alginate-lysozyme protein amphiphilic complex nanoparticle aqueous solution is mixed by a high-speed shearing machine to obtain a water-in-oil-in-water Pickering emulsion loaded with natural active extract of Artemisia selengensis.

[0009] Preferably, the preparation step of the natural active extract solution of Artemisia selengensis in step one is as follows:

[0010] a. After crushing fresh Artemisia selengensis, an ethanol solution with a temperature of 25-100 DEG C and a volume percentage concentration of 70%-100% is added, and the amount of the ethanol solution is 10-250 mL per g of Artemisia selengensis;

[0011] b. Then, the extraction is carried out in a water bath at 25-100 DEG C for 2-120 min or ultrasonic-assisted extraction is carried out to obtain a natural active extract of Artemisia selengensis, which is concentrated under reduced pressure and then spray dried or freeze dried to obtain a dry powder of the natural active extract of Artemisia selengensis, which is stored at -20 DEG C for use;

[0012] c. When used, the dry powder of the natural active extract of Artemisia selengensis is dissolved in water to obtain a natural active extract solution of Artemisia selengensis.

[0013] Preferably, the edible oil is selected from one or a mixture of several of soybean oil, rapeseed oil, peanut oil, corn oil, olive oil, camellia oil, palm oil, sunflower oil, sesame oil, flaxseed oil, and walnut oil.

[0014] Preferably, the preparation method of the sodium alginate polysaccharide / lysozyme protein-based complex nanoparticle solution is as follows:

[0015] Sodium alginate is prepared into a solution with a concentration of 1-2 mg / ml, and lysozyme is prepared into a solution with a concentration of 2-4 mg / ml, and then the two are mixed and stirred by a magnetic stirrer for 30 min to obtain a sodium alginate polysaccharide / lysozyme protein-based complex nanoparticle solution.

[0016] Preferably, the shearing rate of the high-speed shearing machine is 8000-18000 rpm.

[0017] The present application has the following advantages:

[0018] (1) Simple synthesis: the raw materials of the Pickering emulsion of the present application are easy to obtain, the synthesis process is simple, the cost is low, the reaction process is easy to control, and it is suitable for batch production, which is conducive to the popularization and application of commercialization;

[0019] (2) Green and non-toxic: the raw materials for synthesizing the Pickering emulsion of the present application are all food-grade raw materials, and are all green and non-toxic raw materials, so the Pickering emulsion prepared is a green and non-toxic food additive;

[0020] (3) Good antioxidant effect: the Pickering emulsion of the application can effectively prevent the damage of adverse factors such as ultraviolet rays to the natural active extract of Artemisia selengensis;

[0021] (4) Wide application range: the Pickering emulsion of the application has functions such as antioxidant, and can well solve the problem that the natural active substance of Artemisia selengensis is difficult to be applied in the food processing industry due to unstable properties, and is particularly suitable for various functional foods;

[0022] (5) The application utilizes the characteristics of the Pickering emulsion, first, edible oil and lecithin are used to wrap the natural active substance aqueous solution of Artemisia selengensis in a water-in-oil form, and then the sodium alginate-lysozyme protein amphiphilic composite nanoparticles are used to further wrap it, forming a water-in-oil-in-water Pickering emulsion, so that the natural active extract of Artemisia selengensis is protected, thereby improving the stability and bioavailability of the natural active substance of Artemisia selengensis. The Pickering emulsion rich in natural active substance of Artemisia selengensis prepared by the application has more nutritional value compared with the traditional Pickering emulsion, and meets the diversified demand of the market for functional foods. BRIEF DESCRIPTION OF DRAWINGS

[0023] Fig. 1 A, B and C are Artemisia selengensis aqueous solution, soybean oil Artemisia selengensis aqueous solution and water-in-oil-in-water Pickering emulsion loaded with natural active extract of Artemisia selengensis prepared by the application, respectively;

[0024] Fig. 2 A and C are microscope photos of the water-in-oil-in-water Pickering emulsion loaded with natural active extract of Artemisia selengensis under 400 and 800 times magnification field of view, respectively; B and D are fluorescence photos of the Artemisia selengensis natural active extract aqueous solution in each microsphere in the Pickering emulsion under 400 and 800 times magnification field of view, respectively;

[0025] Fig. 3 After 8h irradiation by ultraviolet rays, the flavonoid components in the Artemisia selengensis natural extract embedded in the water-in-oil-in-water Pickering emulsion are well protected, and the residual rate of the flavonoid components is compared with that of the non-embedded group. DETAILED DESCRIPTION

[0026] In order to make the technical means, creative features, purposes and effects realized by the application easy to understand, the application is further described below in combination with specific embodiments.

[0027] For example, Figs. 1 to 3As shown, the Pickering emulsion loaded with Artemisia extract is composed of Artemisia natural active extract aqueous solution, lecithin, edible oil and sodium alginate-lysozyme protein amphiphilic composite nanoparticles.

[0028] The application also discloses a preparation method of the Pickering emulsion loaded with Artemisia extract.

[0029] Step one, after magnetic stirring for 30 min, the Artemisia natural active extract aqueous solution, edible oil and lecithin are prepared into the Artemisia natural active material in the form of water-in-oil, wherein the volume ratio of the Artemisia natural active extract aqueous solution and the edible oil is 1:2, and the amount of lecithin is 3-5 wt% of the mass of the edible oil.

[0030] Step two, the water-in-oil form of the Artemisia natural active material and the equal volume of sodium alginate-lysozyme protein amphiphilic composite nanoparticle aqueous solution are uniformly mixed by a high-speed shearing machine to obtain the Pickering emulsion loaded with Artemisia natural active extract.

[0031] The preparation steps of the Artemisia natural active extract aqueous solution in the step one are as follows:

[0032] a, after crushing, fresh Artemisia is added into an ethanol solution with a temperature of 25-100 DEG C and a volume percentage concentration of 70-100%, and the amount of the ethanol solution is 10-250 mL per g of Artemisia;

[0033] b, then the Artemisia is extracted in a water bath at 25-100 DEG C or ultrasonic-assisted extraction for 2-120 min to obtain the Artemisia natural active extract, which is concentrated under reduced pressure and then spray dried or freeze dried to obtain the dry powder Artemisia natural active extract, which is stored at-20 DEG C for use;

[0034] c, when used, the dry powder Artemisia natural active extract is dissolved in water to obtain the Artemisia natural active extract aqueous solution.

[0035] The edible oil is selected from one or a mixture of several selected from the group consisting of soybean oil, rapeseed oil, peanut oil, corn oil, olive oil, camellia oil, palm oil, sunflower oil, sesame oil, flaxseed oil and walnut oil.

[0036] The preparation method of the sodium alginate polysaccharide / lysozyme protein-based composite nanoparticle solution is as follows:

[0037] Sodium alginate was prepared into a solution with a concentration of 1-2 mg / ml, and lysozyme was prepared into a solution with a concentration of 2-4 mg / ml. After mixing, the sodium alginate polysaccharide / lysozyme protein-based composite nanoparticle solution was obtained by magnetic stirring for 30 minutes. The shearing rate of the high-speed shearing machine was 8000-18000 revolutions per minute.

[0038] Example 1

[0039] Preparation of the water-in-oil-in-water Pickering emulsion loaded with natural active extract of Artemisia rupestris L.: equal volumes of the natural active substance of Artemisia rupestris L. in the form of water-in-oil and the aqueous solution of sodium alginate-lysozyme protein amphiphilic composite nanoparticles were homogenized by a high-speed shearing machine at 10000 rpm to obtain the water-in-oil-in-water Pickering emulsion loaded with natural active extract of Artemisia rupestris L. The emulsion can significantly reduce the damage of ultraviolet light to the related active ingredients in the natural active extract of Artemisia rupestris L. After 8h irradiation by ultraviolet light, the residual rate of flavonoids is 105% higher than that of the unembedded group, and the natural active extract of Artemisia rupestris L. is well protected.

[0040] In the preparation of the natural active extract of Artemisia rupestris L. solution, fresh Artemisia rupestris L. was crushed and added to an ethanol solution with a temperature of 50℃ and a volume percentage concentration of 70%, and the amount of the ethanol solution was 50mL per g of Artemisia rupestris L. Then, the natural active extract of Artemisia rupestris L. was obtained by extraction in a 50℃ water bath or ultrasonic-assisted extraction for 60 minutes. After vacuum concentration, spray drying or freeze drying was performed to obtain dry powder of the natural active extract of Artemisia rupestris L., which was stored at -20℃ for later use. When used, the dry powder of the natural active extract of Artemisia rupestris L. was dissolved in water to obtain the natural active extract of Artemisia rupestris L. solution.

[0041] In the preparation of the sodium alginate-lysozyme protein amphiphilic composite nanoparticles, sodium alginate is a natural anionic polysaccharide, and lysozyme is a natural antibacterial protein with a high isoelectric point. They can be self-assembled into sodium alginate polysaccharide / lysozyme protein-based composite nanoparticles by electrostatic interaction. Sodium alginate was prepared into a solution with a concentration of 2 mg / ml, and lysozyme was prepared into a solution with a concentration of 4 mg / ml. The sodium alginate polysaccharide / lysozyme protein-based composite nanoparticle solution was obtained by magnetic stirring for 30 minutes.

[0042] In the preparation of the natural active substance of Artemisia rupestris L. in the form of water-in-oil, the natural active extract of Artemisia rupestris L. solution and peanut oil were mixed in a ratio of 1:2, and the amount of lecithin was 5wt% of the mass of peanut oil. After magnetic stirring for 30 minutes, the natural active substance of Artemisia rupestris L. in the form of water-in-oil was obtained.

[0043] Example 2:

[0044] Different from the preparation of the water-in-oil-in-water Pickering emulsion loaded with natural active extract of Artemisia rupestris L. in Example 1, the rotation speed of the high-speed shearing machine was 13000 rpm, and the following differences were made:

[0045] In the preparation of the natural active extract of Artemisia rupestris L. aqueous solution, fresh Artemisia rupestris L. was crushed and added to an ethanol solution with a temperature of 75℃ and a volume percentage concentration of 80%, and the amount of the ethanol solution was calculated according to 100 mL of ethanol solution per g of Artemisia rupestris L.; then the natural active extract of Artemisia rupestris L. was obtained by extraction at 75℃ water bath or ultrasonic-assisted extraction for 40 min, and the dry powder of the natural active extract of Artemisia rupestris L. was obtained by spray drying or freeze drying after vacuum concentration; the dry powder of the natural active extract of Artemisia rupestris L. was stored at -20℃ for use; and the natural active extract of Artemisia rupestris L. aqueous solution was obtained by dissolving the dry powder of the natural active extract of Artemisia rupestris L. in water;

[0046] In the preparation of the sodium alginate-lysozyme protein amphiphilic composite nanoparticles, sodium alginate is a natural anionic polysaccharide, and lysozyme is a natural antibacterial protein with high isoelectric point, and the two can be self-assembled into sodium alginate polysaccharide / lysozyme protein-based composite nanoparticles. Sodium alginate was prepared into a solution with a concentration of 1.5 mg / ml, and lysozyme was prepared into a solution with a concentration of 3 mg / ml, and then the sodium alginate polysaccharide / lysozyme protein-based composite nanoparticle solution was obtained by magnetic stirring for 30 minutes;

[0047] In the preparation of the oil-in-water form of natural active substances of Artemisia rupestris L., the natural active extract of Artemisia rupestris L. aqueous solution and walnut oil were mixed in a proportion of 1:2, and the amount of lecithin was 4wt% of the mass of walnut oil, and the oil-in-water form of natural active substances of Artemisia rupestris L. was obtained by magnetic stirring for 30 min.

[0048] The water-in-oil-in-water Pickering emulsion loaded with natural active extract of Artemisia rupestris L. was obtained, which could significantly reduce the damage of ultraviolet rays to the related active ingredients in the natural active extract of Artemisia rupestris L. After 8h irradiation by ultraviolet rays, the residual rate of flavonoids was 123% higher than that of the unembedded group.

[0049] Example 3:

[0050] Different from the preparation of the water-in-oil-in-water Pickering emulsion loaded with natural active extract of Artemisia rupestris L. in Example 1, the rotation speed of the high-speed shearing machine was 13000 rpm, and the following differences were made:

[0051] When the water solution of natural active extract of Artemisia selengensis is prepared, the fresh Artemisia selengensis is crushed, and then added into an ethanol solution with a temperature of 100 DEG C and a volume percentage concentration of 90%, and the amount of the ethanol solution is 200 mL per gram of Artemisia selengensis; then the natural active extract of Artemisia selengensis is obtained by extraction under the condition of a water bath at 100 DEG C for 20 minutes or ultrasonic-assisted extraction; the natural active extract of Artemisia selengensis is obtained by spray drying or freeze drying after concentration under reduced pressure; and the dry powder of the natural active extract of Artemisia selengensis is stored at -20 DEG C for use; and the water solution of the natural active extract of Artemisia selengensis is obtained by dissolving the dry powder of the natural active extract of Artemisia selengensis in water;

[0052] When the sodium alginate-lysozyme protein amphiphilic composite nanoparticles are prepared, the sodium alginate is a natural anionic polysaccharide, and the lysozyme is a natural antibacterial protein with a high isoelectric point; the sodium alginate polysaccharide / lysozyme protein-based composite nanoparticles are obtained by electrostatic self-assembly of the sodium alginate and the lysozyme; the sodium alginate is prepared into a solution with a concentration of 1 mg / ml, and the lysozyme is prepared into a solution with a concentration of 2 mg / ml; and the sodium alginate polysaccharide / lysozyme protein-based composite nanoparticle solution is obtained by magnetic stirring for 30 minutes.

[0053] When the natural active substance of Artemisia selengensis in the form of water-in-oil is prepared, the water solution of the natural active extract of Artemisia selengensis and olive oil are mixed at a ratio of 1:2, and the amount of lecithin is 3 wt% of the mass of the olive oil; and the natural active substance of Artemisia selengensis in the form of water-in-oil is obtained by magnetic stirring for 30 minutes.

[0054] The water-in-oil-in-water Pickering emulsion loaded with the natural active extract of Artemisia selengensis can significantly reduce the damage of ultraviolet rays to the related active ingredients in the natural active extract of Artemisia selengensis; and the residual rate of the flavonoid components is 114% higher than that of the unembedded group after 8 hours of ultraviolet irradiation.

[0055] From the examples 1, 2, 3 and Fig. 3 It can be concluded that the flavonoid components in the natural extract of Artemisia selengensis embedded in the water-in-oil-in-water Pickering emulsion are well protected after 8 hours of ultraviolet irradiation.

[0056] It is to be understood by those skilled in the art that the present application can be implemented by other embodiments without departing from the spirit or essential characteristics thereof. Therefore, the above disclosed embodiments are merely exemplary in all aspects and are not the only ones. All changes within the scope of the present application or within the equivalent scope of the present application are intended to be included in the present application.

Claims

1. A water-in-oil-in-water Pickering emulsion loaded with an extract of Artemisia vulgaris, characterized in that, The Pickering emulsion is composed of an aqueous solution of natural active extract of Artemisia selengensis, lecithin, edible oil and sodium alginate-lysozyme protein amphiphilic composite nanoparticles; The preparation steps of the aqueous solution of natural active extract of Artemisia selengensis are as follows: a. After crushing fresh Artemisia selengensis, add an ethanol solution with a temperature of 25-100℃ and a volume percentage concentration of 70%-100%, the amount of the ethanol solution is 10-250 mL per g of Artemisia selengensis; b. Then, the natural active extract of Artemisia selengensis is obtained by dipping or ultrasonic-assisted dipping for 2-120 min at 25-100℃ in water bath, and the natural active extract of Artemisia selengensis in dry powder form is obtained by spray drying or freeze drying after concentration under reduced pressure, and is stored at 20℃ for later use. 20℃ for later use. c. When used, dissolve the dry powder of natural active extract of Artemisia selengensis in water to obtain an aqueous solution of natural active extract of Artemisia selengensis.

2. The process for the preparation of a water-in-oil-in-water Pickering emulsion loaded with extracts of Artemisia herba-alba according to claim 1, characterized in that: The method comprises the following steps: Step one, after magnetic stirring for 30 min, an oil-in-water form of natural active substance of Artemisia selengensis is prepared from an aqueous solution of natural active extract of Artemisia selengensis, edible oil and lecithin, wherein the volume ratio of the aqueous solution of natural active extract of Artemisia selengensis to the edible oil is 1:2, and the amount of lecithin is 3-5 wt% of the mass of the edible oil; Step two, an oil-in-water-in-water Pickering emulsion loaded with natural active extract of Artemisia selengensis is obtained by homogenously mixing the oil-in-water form of natural active substance of Artemisia selengensis and an aqueous solution of sodium alginate-lysozyme protein amphiphilic composite nanoparticles in equal volumes by using a high-speed shearing machine.

3. The process for the preparation of a water-in-oil-in-water Pickering emulsion loaded with extracts of Artemisia herba-alba according to claim 2, characterized in that: The edible oil is selected from one or a mixture of several of soybean oil, rapeseed oil, peanut oil, corn oil, olive oil, camellia oil, palm oil, sunflower oil, sesame oil, flaxseed oil and walnut oil.

4. The process for the preparation of a water-in-oil-in-water Pickering emulsion loaded with extracts of Artemisia herba-alba according to claim 2, characterized in that: The preparation method of the sodium alginate-lysozyme protein amphiphilic composite nanoparticle solution is as follows: Prepare a 1-2 mg / ml solution of sodium alginate and a 2-4 mg / ml solution of lysozyme, mix them and then stir magnetically for 30 min to obtain a sodium alginate-lysozyme protein amphiphilic composite nanoparticle solution.

5. The process for the preparation of a water-in-oil-in-water Pickering emulsion loaded with extracts of Artemisia herba-alba according to claim 2, characterized in that: The shearing rate of the high-speed shearing machine is 8000-18000 revolutions per minute.

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