阅读说明：本技术 一种干细胞外泌体冻干粉溶媒制备方法 (Preparation method of stem cell exosome freeze-dried powder solvent ) 是由 武丽芳 马小梅 邢梦洁 张红钢 于 2021-08-19 设计创作，主要内容包括：本发明公开了一种干细胞外泌体冻干粉溶媒制备方法,其技术方案要点是：包括以下步骤：步骤一、准备原料,按照质量份数取神经酰胺10-15份、甘油2-5份、维生素4-7份、皮肤促渗剂2-5份、抗氧化剂2-5份、植物提取物4-7份、保湿剂2-5份、寡肽4-7份、苯乙酮3-6份、丙二醇4-7份和去离子水；步骤二、一次材料混合,将神经酰胺、甘油、植物提取物、维生素和去离子水按照重量份数投入到混合桶内,通过搅拌器进行搅拌,使得各给原料之间充分混合,有利于提高原料之间的充分融合效率,流程明确以及清晰,通过制备方法中加入的皮肤促渗剂有利于提高原料成分透过表皮到达真皮层的速度,提高护肤的效果,保持溶媒中的各个材料的优良性能。(The invention discloses a preparation method of a stem cell exosome freeze-dried powder solvent, which has the technical scheme key points that: the method comprises the following steps: preparing raw materials, namely taking 10-15 parts of ceramide, 2-5 parts of glycerol, 4-7 parts of vitamin, 2-5 parts of skin penetration enhancer, 2-5 parts of antioxidant, 4-7 parts of plant extract, 2-5 parts of humectant, 4-7 parts of oligopeptide, 3-6 parts of acetophenone, 4-7 parts of propylene glycol and deionized water according to parts by mass; and step two, mixing the materials once, namely putting ceramide, glycerol, the plant extract, vitamins and deionized water into a mixing barrel according to parts by weight, stirring by a stirrer to ensure that the raw materials are fully mixed, so that the full fusion efficiency of the raw materials is improved, the process is clear and clear, and the skin penetration enhancer added in the preparation method is favorable for improving the speed of the raw material components penetrating through the epidermis to reach the dermis layer, the skin care effect is improved, and the excellent performance of each material in the solvent is kept.)

1. A preparation method of a stem cell exosome freeze-dried powder solvent is characterized by comprising the following steps: the method comprises the following steps:

preparing raw materials, namely taking 10-15 parts of ceramide, 2-5 parts of glycerol, 4-7 parts of vitamin, 2-5 parts of skin penetration enhancer, 2-5 parts of antioxidant, 4-7 parts of plant extract, 2-5 parts of humectant, 4-7 parts of oligopeptide, 3-6 parts of acetophenone, 4-7 parts of propylene glycol and deionized water according to parts by mass;

step two, mixing the primary materials, namely putting ceramide, glycerol, plant extracts, vitamins and deionized water into a mixing barrel according to parts by weight, and stirring by a stirrer to fully mix the raw materials;

step three, heating and stirring, namely putting the fully mixed raw materials in the step into a reaction skin penetration enhancer kettle for heating and stirring by a stirrer, wherein the heating time is 20-40 minutes until the raw materials in the reaction kettle are fully dissolved to obtain a reactant A;

step four, heating in a water bath, namely putting the oligopeptide, the acetophenone and the propylene glycol into a water bath container according to parts by weight, mixing and heating in the water bath, uniformly stirring and heating by a stirrer until the oligopeptide, the acetophenone and the propylene glycol are fully dissolved to obtain a mixture B;

step five, mixing the secondary materials, reducing the temperature in the reaction kettle to 50-65 ℃, putting the mixture B obtained in the step four into the reaction kettle, fully stirring the mixture B and the reactant A, and fully fusing to obtain a mixture C;

mixing the materials in the sixth step and the third step, adding the skin penetration enhancer, the antioxidant and the humectant into the reaction kettle according to the parts by weight, and fully stirring the mixture C for 16-24 minutes to obtain a solvent liquid;

and seventhly, cooling, namely cooling the solvent liquid through cooling equipment, and filtering through a filter screen.

2. The method for preparing a lyophilized powder solvent of a stem cell exosome according to claim 1, which is characterized in that: the antioxidant is one or more of tocopherol antioxidant, butylated hydroxyanisole antioxidant, butylated hydroxytoluene antioxidant and tertiary butyl hydroquinone.

3. The method for preparing a lyophilized powder solvent of a stem cell exosome according to claim 1, which is characterized in that: the plant extract is one or more of seaweed extract, tea extract, crocus sativus extract and centella asiatica extract.

4. The method for preparing a lyophilized powder solvent of a stem cell exosome according to claim 1, which is characterized in that: the humectant is one or more of a sodium PCA humectant, a sodium lactate humectant and a hydroxyethyl urea humectant.

5. The method for preparing a lyophilized powder solvent of a stem cell exosome according to claim 1, which is characterized in that: the oligopeptide is one or more of oligopeptide-1, oligopeptide-3, oligopeptide-5 and oligopeptide-6.

6. The method for preparing a lyophilized powder solvent of a stem cell exosome according to claim 1, which is characterized in that: the vitamin is one or more of vitamin A, vitamin D, vitamin E and vitamin K.

7. The method for preparing a lyophilized powder solvent of a stem cell exosome according to claim 1, which is characterized in that: the stirring speed of the stirrer in the second step is 300-400 rpm, and the stirring time is 10-25 minutes.

8. The method for preparing a lyophilized powder solvent of a stem cell exosome according to claim 1, which is characterized in that: and in the third step, the reaction kettle is heated to 75-90 ℃, and the stirring time of the stirrer is 20-25 minutes.

9. The method for preparing a lyophilized powder solvent of a stem cell exosome according to claim 1, which is characterized in that: in the fourth step, the temperature of the water bath is 50-65 ℃, and the heating time is 30-40 minutes.

10. The method for preparing a lyophilized powder solvent of a stem cell exosome according to claim 1, which is characterized in that: and in the seventh step, the temperature of the solvent liquid is reduced to 21-27 ℃, the cooling time is 4-5 hours, and the filter screen is a filter screen with the aperture of 0.5-1.5 mu m.

Technical Field

The invention relates to the technical field of freeze-dried powder solvents, and in particular relates to a preparation method of a stem cell exosome freeze-dried powder solvent.

Background

The lyophilized powder is sterile powder injection prepared by freezing the medicinal liquid into solid in sterile environment, and vacuum-pumping to sublimate and dry water. Effectively prevents the change of the physicochemical and biological characteristics of the product, has small damage to the structure and the characteristics of biological tissues and cells, leads the biological tissues and the cells to rapidly enter a dormant state, and effectively protects the stability of the effective components of a plurality of thermosensitive drug biological products. Such as protein, microbe, etc. without denaturation and loss of bioactivity; secondly, the freeze-dried product is loose in shape and basically not changed in color after being dried, and can be quickly dissolved and recover the physical and chemical properties and biological activity of the original aqueous solution after being added with water. Thirdly, because the drying is carried out under the vacuum condition, the protective effect on some substances which are easy to oxidize is good. Fourthly, the water content of the freeze-dried product is very low, so that the stability of the product is improved, the chance of pollution is reduced, the transportation is convenient, the storage life of the product is prolonged, and the solvent is used for dissolving the freeze-dried powder.

For example, chinese patent No. CN111388394A discloses a two-phase lyophilized powder solvent, and a preparation method and application thereof. The two-phase freeze-dried powder solvent comprises an oil phase component and a water phase component, wherein the oil phase component comprises ceramide, phytosterol oleate, wild soybean oil, an antioxidant and base oil.

The freeze-dried powder solvent has the advantages of enhancing the water locking capacity and the repairing capacity of the skin; however, the above-mentioned lyophilized powder solvent still has some disadvantages, such as: the preparation process is simple and crude, and the penetration of the solvent to the skin is poor.

Disclosure of Invention

The invention aims to provide a preparation method of a stem cell exosome freeze-dried powder solvent, and aims to solve the problems in the background technology.

In order to achieve the purpose, the invention provides the following technical scheme:

a preparation method of a stem cell exosome freeze-dried powder solvent comprises the following steps:

preparing raw materials, namely taking 10-15 parts of ceramide, 2-5 parts of glycerol, 4-7 parts of vitamin, 2-5 parts of skin penetration enhancer, 2-5 parts of antioxidant, 4-7 parts of plant extract, 2-5 parts of humectant, 4-7 parts of oligopeptide, 3-6 parts of acetophenone, 4-7 parts of propylene glycol and deionized water according to parts by mass;

step two, mixing the primary materials, namely putting ceramide, glycerol, plant extracts, vitamins and deionized water into a mixing barrel according to parts by weight, and stirring by a stirrer to fully mix the raw materials;

step three, heating and stirring, namely putting the fully mixed raw materials in the step into a reaction skin penetration enhancer kettle for heating and stirring by a stirrer, wherein the heating time is 20-40 minutes until the raw materials in the reaction kettle are fully dissolved to obtain a reactant A;

step four, heating in a water bath, namely putting the oligopeptide, the acetophenone and the propylene glycol into a water bath container according to parts by weight, mixing and heating in the water bath, uniformly stirring and heating by a stirrer until the oligopeptide, the acetophenone and the propylene glycol are fully dissolved to obtain a mixture B;

step five, mixing the secondary materials, reducing the temperature in the reaction kettle to 50-65 ℃, putting the mixture B obtained in the step four into the reaction kettle, fully stirring the mixture B and the reactant A, and fully fusing to obtain a mixture C;

mixing the materials in the sixth step and the third step, adding the skin penetration enhancer, the antioxidant and the humectant into the reaction kettle according to the parts by weight, and fully stirring the mixture C for 16-24 minutes to obtain a solvent liquid;

and seventhly, cooling, namely cooling the solvent liquid through cooling equipment, and filtering through a filter screen.

Preferably, the antioxidant is one or more of tocopherol antioxidant, butylated hydroxyanisole antioxidant, butylated hydroxytoluene antioxidant and tertiary butyl hydroquinone.

Preferably, the plant extract is one or more of seaweed extract, tea extract, saffron extract and centella extract.

Preferably, the moisturizer is one or more of a sodium PCA moisturizer, a sodium lactate moisturizer, and a hydroxyethyl urea moisturizer.

Preferably, the oligopeptide is one or more of oligopeptide-1, oligopeptide-3, oligopeptide-5 and oligopeptide-6.

Preferably, the vitamin is one or more of vitamin a, vitamin D, vitamin E and vitamin K.

Preferably, the stirring speed of the stirrer in the second step is 300-400 rpm, and the stirring time is 10-25 minutes.

Preferably, the reaction kettle in the third step is heated to 75-90 ℃, and the stirring time of the stirrer is 20-25 minutes.

Preferably, the water bath temperature in the fourth step is 50-65 ℃, and the heating time is 30-40 minutes.

Preferably, in the seventh step, the temperature of the solvent liquid is reduced to 21-27 ℃, the cooling time is 4-5 hours, and the filter screen is a filter screen with the aperture of 0.5-1.5 μm.

Compared with the prior art, the invention has the beneficial effects that:

the steps of primary material mixing, secondary material mixing, tertiary material mixing and the like in the preparation method of the dry cell exosome freeze-dried powder solvent are favorable for improving the sufficient fusion efficiency among raw materials, the flow is clear and definite, the speed of the raw material components reaching the dermis layer through the epidermis is favorably improved through the skin penetration enhancer added in the preparation method, the skin care effect is improved, the storage time of the solvent is favorably prolonged through the antioxidant, the excellent performance of each material in the solvent can be kept, the service life of the solvent is prolonged, the moisturizing effect and the moisturizing time of the solvent are favorably improved through the provided humectant, collagen is favorably and rapidly supplemented through oligopeptide, the metabolism is promoted, and the regeneration capacity of the collagen is favorably realized.

Drawings

FIG. 1 is a block flow diagram of the present invention.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1

A preparation method of a stem cell exosome freeze-dried powder solvent comprises the following steps:

preparing raw materials, namely taking 10 parts of ceramide, 2 parts of glycerol, 4 parts of vitamin, 2 parts of skin penetration enhancer, 2 parts of antioxidant, 4 parts of plant extract, 2 parts of humectant, 4 parts of oligopeptide, 3 parts of acetophenone, 4 parts of propylene glycol and deionized water according to the mass parts;

step two, mixing the primary materials, namely putting ceramide, glycerol, plant extracts, vitamins and deionized water into a mixing barrel according to parts by weight, and stirring by a stirrer to fully mix the raw materials;

step three, heating and stirring, namely putting the fully mixed raw materials in the step into a reaction skin penetration enhancer kettle for heating and stirring by a stirrer, wherein the heating time is 20 minutes until the raw materials in the reaction kettle are fully dissolved to obtain a reactant A;

step four, heating in a water bath, namely putting the oligopeptide, the acetophenone and the propylene glycol into a water bath container according to parts by weight, mixing and heating in the water bath, uniformly stirring and heating by a stirrer until the oligopeptide, the acetophenone and the propylene glycol are fully dissolved to obtain a mixture B;

step five, mixing the secondary materials, reducing the temperature in the reaction kettle to 50 ℃, putting the mixture B obtained in the step four into the reaction kettle, fully stirring the mixture B and the reactant A, and fully fusing to obtain a mixture C;

mixing the materials in the sixth step and the third step, adding the skin penetration enhancer, the antioxidant and the humectant into the reaction kettle according to the parts by weight, and fully stirring the mixture C for 16 minutes to obtain a solvent liquid;

and seventhly, cooling, namely cooling the solvent liquid through cooling equipment, and filtering through a filter screen.

In this embodiment, the antioxidant is preferably a tocopherol antioxidant and a butylated hydroxyanisole antioxidant.

In this embodiment, the plant extract is preferably an alga extract or a tea extract.

In this embodiment, the humectants are preferably sodium PCA humectants and sodium lactate humectants.

In this embodiment, the oligopeptide is preferably oligopeptide-1 and oligopeptide-3.

In this embodiment, the vitamins are preferably vitamin a and vitamin D.

In this embodiment, the stirring speed of the stirrer in the second step is preferably 300 rpm, and the stirring time is preferably 10 minutes.

In this embodiment, preferably, in the third step, the reaction kettle is heated to 75 ℃, and the stirring time of the stirrer is 20 minutes.

In this embodiment, preferably, in the fourth step, the water bath temperature is 50 ℃, and the heating time is 30 minutes.

In this embodiment, preferably, in the seventh step, the temperature of the solvent liquid is reduced to 21 ℃, the time of the temperature reduction is 4 hours, and the filter screen is a filter screen with a pore size of 0.5 μm.

Example 2

A preparation method of a stem cell exosome freeze-dried powder solvent comprises the following steps:

preparing raw materials, namely taking 12 parts of ceramide, 3 parts of glycerol, 6 parts of vitamin, 4 parts of skin penetration enhancer, 3 parts of antioxidant, 6 parts of plant extract, 4 parts of humectant, 6 parts of oligopeptide, 4 parts of acetophenone, 5 parts of propylene glycol and deionized water according to the mass parts;

step two, mixing the primary materials, namely putting ceramide, glycerol, plant extracts, vitamins and deionized water into a mixing barrel according to parts by weight, and stirring by a stirrer to fully mix the raw materials;

step three, heating and stirring, namely putting the fully mixed raw materials in the step into a reaction skin penetration enhancer kettle for heating and stirring by a stirrer, wherein the heating time is 30 minutes until the raw materials in the reaction kettle are fully dissolved to obtain a reactant A;

step four, heating in a water bath, namely putting the oligopeptide, the acetophenone and the propylene glycol into a water bath container according to parts by weight, mixing and heating in the water bath, uniformly stirring and heating by a stirrer until the oligopeptide, the acetophenone and the propylene glycol are fully dissolved to obtain a mixture B;

step five, mixing the secondary materials, cooling the temperature in the reaction kettle to 60 ℃, putting the mixture B obtained in the step four into the reaction kettle, fully stirring the mixture B and the reactant A, and fully fusing to obtain a mixture C;

mixing the materials in the sixth step and the third step, adding the skin penetration enhancer, the antioxidant and the humectant into the reaction kettle according to the parts by weight, and fully stirring the mixture C for 20 minutes to obtain a solvent liquid;

and seventhly, cooling, namely cooling the solvent liquid through cooling equipment, and filtering through a filter screen.

In this embodiment, the antioxidant is preferably tert-butylhydroquinone.

In this embodiment, the plant extract is preferably an extract of centella asiatica.

In this embodiment, preferably, the humectant is a hydroxyethyl urea humectant.

In this embodiment, the oligopeptide is preferably oligopeptide-6.

In this embodiment, the vitamins are preferably vitamin E and vitamin K.

In this embodiment, the stirring speed of the stirrer in the second step is preferably 350 rpm, and the stirring time is preferably 15 minutes.

In this embodiment, preferably, in the third step, the reaction kettle is heated to 85 ℃ and the stirring time of the stirrer is 24 minutes.

In this embodiment, preferably, in the fourth step, the water bath temperature is 60 ℃, and the heating time is 35 minutes.

In this embodiment, preferably, in the seventh step, the temperature of the solvent liquid is reduced to 25 ℃, the time of the temperature reduction is 4.5 hours, and the filter screen is a filter screen with a pore size of 1 μm.

Example 3

A preparation method of a stem cell exosome freeze-dried powder solvent comprises the following steps:

preparing raw materials, namely taking 15 parts of ceramide, 5 parts of glycerol, 7 parts of vitamin, 5 parts of skin penetration enhancer, 5 parts of antioxidant, 7 parts of plant extract, 5 parts of humectant, 7 parts of oligopeptide, 6 parts of acetophenone, 7 parts of propylene glycol and deionized water according to the mass parts;

step two, mixing the primary materials, namely putting ceramide, glycerol, plant extracts, vitamins and deionized water into a mixing barrel according to parts by weight, and stirring by a stirrer to fully mix the raw materials;

step three, heating and stirring, namely putting the fully mixed raw materials in the step into a reaction skin penetration enhancer kettle for heating and stirring by a stirrer, wherein the heating time is 40 minutes until the raw materials in the reaction kettle are fully dissolved to obtain a reactant A;

step four, heating in a water bath, namely putting the oligopeptide, the acetophenone and the propylene glycol into a water bath container according to parts by weight, mixing and heating in the water bath, uniformly stirring and heating by a stirrer until the oligopeptide, the acetophenone and the propylene glycol are fully dissolved to obtain a mixture B;

step five, mixing the secondary materials, reducing the temperature in the reaction kettle to 65 ℃, putting the mixture B obtained in the step four into the reaction kettle, fully stirring the mixture B and the reactant A, and fully fusing to obtain a mixture C;

mixing the materials in the sixth step and the third step, adding the skin penetration enhancer, the antioxidant and the humectant into the reaction kettle according to the parts by weight, and fully stirring the mixture C for 24 minutes to obtain a solvent liquid;

and seventhly, cooling, namely cooling the solvent liquid through cooling equipment, and filtering through a filter screen.

In this embodiment, the antioxidant is preferably a tocopherol antioxidant and a dibutylhydroxytoluene antioxidant.

In this embodiment, the plant extracts are preferably saffron extract and centella asiatica extract.

In this embodiment, the moisturizers are preferably sodium lactate moisturizer and hydroxyethyl urea moisturizer.

In this embodiment, the oligopeptide is preferably oligopeptide-3 and oligopeptide-5.

In this embodiment, preferably, the vitamin is vitamin a.

In this embodiment, the stirring speed of the stirrer in the second step is preferably 400 rpm, and the stirring time is preferably 5 minutes.

In this embodiment, preferably, in the third step, the reaction kettle is heated to 90 ℃, and the stirring time of the stirrer is 25 minutes.

In this embodiment, preferably, the water bath temperature in the fourth step is 65 ℃, and the heating time is 40 minutes.

In this embodiment, preferably, in the seventh step, the temperature of the solvent liquid is reduced to 27 ℃, the time of the temperature reduction is 5 hours, and the filter screen is a filter screen with a pore size of 1.5 μm.

Skin feel test;

40 subjects were selected and randomized into 4 groups of 10 persons each and samples of example 1, example 2 and example 3 were tested against comparative example 1 and comparative example 2.

The specific method comprises the following steps: cleaning the surface of an arm with clear water, taking a proper amount of sample, uniformly applying the sample on the surface of the skin of the arm, and evaluating the absorption speed, the freshness, the moisturizing effect, the moistening effect, the softness and the tenderness of the freeze-dried powder according to the feeling in the using process and 30 minutes after the use. The scoring results are shown in table 1, each scoring range is 1-10 points, and the higher the score is, the better the effect of the sample is.

It is understood from the table that the absorption rate, the degree of freshness, the moisturizing effect, the softness and the tenderness score in example 3 are high.

The working principle and the using process of the invention are as follows:

the steps of primary material mixing, secondary material mixing, tertiary material mixing and the like in the preparation method of the dry cell exosome freeze-dried powder solvent are favorable for improving the sufficient fusion efficiency among raw materials, the flow is clear and definite, the speed of the raw material components reaching the dermis layer through the epidermis is favorably improved through the skin penetration enhancer added in the preparation method, the skin care effect is improved, the storage time of the solvent is favorably prolonged through the antioxidant, the excellent performance of each material in the solvent can be kept, the service life of the solvent is prolonged, the moisturizing effect and the moisturizing time of the solvent are favorably improved through the provided humectant, collagen is favorably and rapidly supplemented through oligopeptide, the metabolism is promoted, and the regeneration capacity of the collagen is favorably realized.

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.