CN113679647A

CN113679647A - Composition containing long-hair dioscorea root extract and preparation and application thereof

Info

Publication number: CN113679647A
Application number: CN202110960506.7A
Authority: CN
Inventors: 俞文洁
Original assignee: Individual
Current assignee: Individual
Priority date: 2021-08-20
Filing date: 2021-08-20
Publication date: 2021-11-23

Abstract

The invention discloses a composition containing an extract of dioscorea villosa root, comprising: polyhydric alcohol, lecithin and extract of long hair yam root. The weight percentage of the polyhydric alcohol is 60-90%, the weight percentage of the lecithin is 1-15%, and the weight percentage of the long-hair dioscorea root extract is 0.1-8%. The composition of the present invention containing the extract of the roots of dioscorea villosa may further include fish egg protein. The invention also relates to the preparation of the composition containing the long-hair dioscorea root extract and the application of the composition in cosmetics.

Description

Composition containing long-hair dioscorea root extract and preparation and application thereof

Technical Field

The invention relates to a composition and application thereof, belongs to the field of cosmetics, and particularly relates to preparation of a composition of a long-hair dioscorea root extract and fish egg protein and application of the composition in a skin external preparation.

Background

As women age, especially women in climacteric period, and their gland secretion is unbalanced, such as thyroid gland, pancreas, gonad, adrenal gland, pituitary gland, pineal gland, etc., the effect of estrogen level decrease is more obvious on facial skin and body skin health. These effects include wrinkles, dryness, thinning of the skin, reduced collagen content, slow healing of wounds, loss of elasticity, sagging and the like. The Estrogen can be supplemented to increase the thickness, elasticity and collagen quantity of the skin, so that cells can be actively divided, the damaged skin can be repaired, the blood supply can be increased, the skin shows that the action mechanism of the human-activating halation … … has a great relationship with Estrogen Receptor (ER) protein, the human-activating halation … … belongs to GPER1(GPR30), Gap-ER and ER-X of a G protein coupled protein receptor family, the human-activating halation … …, the Gap-ER and the ER-X mediate quick non-genotype effects, play an indirect transcription regulation and control role through a second messenger system, and participate in multifunctional regulation of skin cell activation, regeneration, injury repair and the like. The estrogen receptor protein exists in cell membranes, cytoplasm and cell nucleus, is very sensitive, and when skin is irradiated by ultraviolet rays, the estrogen receptor protein is dormant to cause the obstruction of an estrogen signal channel, thereby being one of the main reasons for generating photoaging.

In addition, studies on impaired growth hormone secretion of adult people, published in "J.Clin.invest", Daniel R, and the like, have shown that with the increase of age, estrogen receptor protein is gradually weakened, and the amount of hormone secreted by the body is obviously reduced with the increase of age, so that stimulation of estrogen signals is hindered, and various problems of the body are generated.

The extract of Dioscorea hirta root, Dioscorea Villosa, Latin name Dioscorea Villosa, is extracted from the rhizome of a perennial vine plant of Dioscoreaceae. The plant is characterized in that the woody rootstock (root tuber) is thick, long, thin, bent and multi-node, and the variety produced in Mexico areas has the best quality. The National Institutes of Health (NIH) studies have shown that saponin molecules extracted from the roots of Dioscorea villosa are very similar in molecular structure to human native lutein, and that these saponin molecules have a trisaccharide glycoside consisting of two L-rhamnose and one D-glucose molecule, or two D-glucose and one L-rhamnose molecule. After being absorbed by skin, the saponin molecule can be used as a natural lutein similar component to activate hormone required by the secretion of human tissues, thereby regulating the balance of the hormone in the human body.

The roe protein is a protein taken from eggs of salmon (Oncorhynchus tschawytscha), trout (Squaliobarbus curriculus), herring (clupearia pallasi) and the like, is a cationic active peptide of basic amino acid, mainly comprises arginine and lysine according to a certain sequence, and the amino acid contains guanidino (PKa ═ 12.48), so that the roe protein carries a large number of positive charges and can interact with phospholipid carrying negative charges on cell membranes or muramic acid carrying negative charges on cell walls, thereby improving the permeability of the cell membranes or the cell walls and having membrane translocation and membrane transport functions. The sulfate of the fish egg protein adopted by the invention is also called protamine sulfate and protamine sulfate. Strongly basic protein sulfate isolated from male fish germ cells or milt.

In the prior art, chinese patent document 201910828987.9 discloses an anti-aging eye cream of dioscorea hispida and a preparation method thereof. It discloses only an eye cream containing an extract of dioscorea hispida roots. The research does not find the application of the combination of the dioscorea villosa root extract and the fish egg protein in the cosmetic field.

In order to fully exert the effect of the dioscorea zingiberensis root extract on the skin, improve the bioavailability of the dioscorea zingiberensis root extract and protect the stability of the active ingredients of the dioscorea zingiberensis root, the invention unexpectedly discovers a microcapsule structure, which is coated layer by layer in an onion mode, and anchors fish egg protein on the surface of the microcapsule so as to improve the permeability of active substances to the skin and maintain the long-time release characteristic of the active substances in the skin. The composition of the roe protein combined with the dioscorea villosa root extract microcapsule can change the permeability of cell membranes and greatly improve the transdermal absorption of skin, the roe protein transdermal peptide combines the microcapsule with estrogen receptor protein to realize strong stimulation on the estrogen receptor protein and provide a large dose of lutein, and the microcapsules are released layer by layer to improve the state of human estrogen; automatically and bidirectionally regulating hormone required by human body to balance state, opening blood stasis blocking lactic acid nodule, discharging turbid substance in vivo, activating adolescent energy, regulating endocrine, and recovering health and vitality.

A precursor similar to lutein of a human body is obtained from the long-hair dioscorea root extract, the precursor is absorbed through the skin, the burden of the liver caused by an internal administration mode is avoided, and the long-hair dioscorea root rich in plant hormone precursor substances is delivered to the action part of the skin through continuously optimizing the proportion. The microcapsule structure in the invention has extremely excellent affinity with skin, excellent active matter bearing and penetration, and the long-hair dioscorea root extract is not hormone and has no side effect.

Disclosure of Invention

In one aspect, the present invention provides a composition containing an extract of dioscorea villosa root, comprising: polyhydric alcohol, lecithin and extract of long hair yam root.

In a preferred embodiment, the composition of the present invention comprises 60 to 90% by mass of the polyhydric alcohol.

In a preferred embodiment, the polyhydric alcohol in the composition containing the extract of the long dioscorea villosa root of the present invention is selected from any two of glycerin, butylene glycol, pentylene glycol, and 1, 3-propanediol.

In a preferred embodiment, the composition of the present invention comprises 3.3 to 12% by mass of lecithin in the extract from the root of dioscorea villosa.

In a preferred embodiment, the composition containing the long dioscorea villosa root extract comprises 3.3 to 8% of the long dioscorea villosa root extract by mass.

In another aspect, the present invention relates to a method for preparing a composition containing an extract of dioscorea villosa root, comprising the steps of:

1a) dispersing a long dioscorea villosa root extract and lecithin in a polyol to form a polyol system;

1b) lyophilizing or homogenizing under high pressure to obtain composition containing extract of Dioscorea villosa root;

in a preferred embodiment, the freeze-drying is arranged to: sublimation drying is carried out at the temperature of-80 to-40 ℃ and the vacuum degree of 1 to 30Pa for 3 to 21 hours, desorption drying is carried out at the temperature of-10 to +20 ℃ and the vacuum degree of 1 to 30Pa for 1 to 10 hours, and the water content is controlled to be 0 to 6 percent.

In a preferred embodiment, the spray drying is arranged to: the air inlet temperature is 40-80 ℃, the atomization pressure is 0.2-0.8 MPa, the feeding speed is 100-2000 ml/h, and the water content is controlled to be 0-8%.

In a preferred embodiment, the high pressure homogenization is arranged to: the circulation is carried out for 3 times at the flow rate of 1200L/h and the pressure of 100 MPa.

In still another aspect, the present invention relates to a skin external preparation comprising an extract of dioscorea villosa root and fish egg protein.

In a preferred embodiment, the skin external preparation comprises an extract of dioscorea villosa root and fish egg protein selected from the group consisting of: salmon protamine, trout protamine, herring protamine.

In a preferred embodiment, the skin external preparation comprises the extract of the root of dioscorea villosa and the roe protein, wherein the mass percentage of the roe protein is less than or equal to 16.7 percent.

In still another aspect, the present invention also relates to a method for preparing a skin external preparation comprising an extract of dioscorea villosa root and roe protein, the method comprising the steps of:

1c) mixing the fish egg protein with the composition comprising the extract of the roots of dioscorea villosa prepared in step 1b) to obtain the skin external preparation comprising the fish egg protein.

In yet another aspect, the present invention relates to a composition comprising an extract of the root of dioscorea villosa, for use in cosmetics.

In still another aspect, the present invention relates to a skin external preparation comprising an extract from the root of dioscorea villosa and fish egg protein, for use in cosmetics.

Drawings

Fig. 1 shows an electron micrograph of the composition of example 1 comprising dioscorea villosa root extract, lecithin, polyol, and fish egg protein.

Detailed Description

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are described herein. For the purposes of the present invention, the following terms are defined below.

The term "about" as used herein refers to an amount, level, value, dimension, size, or amount that differs by up to 30%, 20%, or 10% as compared to the amount, level, value, dimension, size, or amount of a reference. The percentages used herein are by weight unless otherwise indicated.

Throughout the specification and claims, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

The present invention is based on the following unexpected findings: the composition containing Dioscorea villosa root extract can be used in combination with fish egg protein to prepare stable skin external preparation. Moreover, the skin external preparation containing the dioscorea villosa root extract composition and the fish egg protein can remarkably improve the intake of the dioscorea villosa root extract, thereby improving the bioavailability of the dioscorea villosa root extract.

The inventors of the present application have also found that in order to fully exert the effect of the dioscorea villosa root extract on the skin, improve the bioavailability thereof, and protect the stability of the active ingredient of dioscorea villosa root, the present invention has unexpectedly found a microcapsule structure, which is coated with "onion" type layer by layer, and anchors the fish egg protein on the surface of the microcapsule, so as to improve the permeability of the active substance to the skin and maintain the long-term release characteristics of the active substance in the skin. The composition of the roe protein combined with the dioscorea villosa root extract microcapsule can change the permeability of cell membranes and greatly improve the transdermal absorption of skin, the roe protein transdermal peptide combines the microcapsule with estrogen receptor protein to realize strong stimulation on the estrogen receptor protein and provide a large dose of lutein, and the microcapsules are released layer by layer to improve the state of human estrogen; automatically and bidirectionally regulating hormone required by human body to balance state, opening blood stasis blocking lactic acid nodule, discharging turbid substance in vivo, activating adolescent energy, regulating endocrine, and recovering health and vitality.

External preparation for skin

The skin external preparation of the present invention comprises a long-hair dioscorea root extract, lecithin, a polyol, and fish egg protein, and has an onion-type layer-by-layer encapsulated microcapsule structure. Here, the external preparation for skin may be referred to as an external preparation for skin composition or an external composition for skin. In the present invention, the "onion-type layer-by-layer microcapsule structure" means a structure in which fluidity is maintained while regularity of molecular arrangement in which a bimolecular film of an amphiphilic molecule and water are alternately oriented is retained. Therefore, the term "having an" onion-type layer-by-layer microcapsule structure "means that the skin preparation for external use contains a lamellar microcapsule structure, and the lamellar microcapsule structure can be confirmed, for example, by using a polarization microscope according to the method and conditions described in examples described later.

The invention also aims to provide the application of the composition in preparing a skin external preparation.

Preferably, the skin external preparation is a cosmetic, preferably a lotion, essence, mask, emulsion, cream;

preferably, the content of the composition in the skin external preparation is 0.001 to 50% by mass, for example, 0.005%, 0.01%, 0.05%, 0.1%, 0.5%, 1.5%, 3%, 5%, 12%, 22%, 30%, 35%, 40%, 45%, 49% or the like, preferably 1 to 20%, more preferably 2 to 10%, most preferably 3 to 8%.

The composition prepared by combining the specific fish egg protein can obviously accelerate the skin permeability of the dioscorea villosa root extract, increase the release time of the dioscorea villosa root extract in the skin, and has excellent safety, stability and efficacy; compared with the composition which is obtained by the same components and preparation process and is not added with the fish egg protein, the composition or the skin external agent containing the fish egg protein has obvious advantages.

Extract of Dioscorea villosa root

Fish egg protein

Lecithin

The lecithin used in the present invention is a lecithin derived from soybean or egg yolk and an enzyme-treated lecithin, and may be subjected to chemical treatment such as hydrogenation. Here, lecithin is a general term for a mixture mainly composed of various phospholipids extracted from animals and plants, and mainly composed of Phosphatidylcholine (PC), Phosphatidylethanolamine (PE), Phosphatidylinositol (PI), and Phosphatidic Acid (PA). As the lecithin used in the present invention, a complex (lecithin complex) obtained by combining a sterol and lecithin can be used. Examples of such a complex include a complex of hydrogenated lecithin and phytosterol, and a complex of hydrogenated lecithin and cholesterol. Therefore, as the lecithin to be used, a complex obtained by combining a sterol and lecithin is preferably used. The content of lecithin in the external preparation for skin of the present invention is not particularly limited, and may be appropriately adjusted depending on the form and purpose of the external preparation for skin, the composition ratio of other components, and the like. For example, the content of lecithin is preferably 0.1 to 15% by mass, more preferably 3.3 to 12% by mass, based on the whole skin preparation for external use.

Polyhydric alcohols

The polyol that can be used in the present invention is not particularly limited as long as it is a water-soluble polyol having 2 or more hydroxyl groups in the molecule. Specific examples thereof include glycerin, diglycerin, triglycerin, polyethylene glycol, propylene glycol, 1, 3-butanediol, 1, 2-pentanediol, and sorbitol. Among them, glycerin, 1, 3-butanediol, and 1, 2-pentanediol are preferably used in view of excellent stability of the lamellar liquid crystal structure. These polyols may be used alone, or in combination of 2 or more. The content of the polyhydric alcohol in the skin external preparation of the present invention is not particularly limited, and may be appropriately adjusted depending on the form and purpose of the skin external preparation, the composition ratio of other components, and the like. For example, the amount is preferably 60 to 90% by mass, and more preferably 65 to 86% by mass, based on the whole skin preparation for external use.

The content of the fish egg protein in the external preparation for skin of the present invention is not particularly limited, and may be appropriately adjusted depending on the form and purpose of the external preparation for skin, the composition ratio of other components, and the like. For example, the amount is preferably 0.001 to 20% by mass, more preferably 0.01 to 10% by mass, based on the whole skin preparation for external use. In general, the content of the egg protein is 0.001% by mass or more, and it is considered that the white, porous and spongy skin preparation for external use is stabilized by the surface activity of the egg protein, and the stability of the skin preparation for external use is advantageously maintained.

According to the study of the present invention, it was found that fish egg protein has an interaction with lecithin and a polyhydric alcohol, and fish egg protein can improve the affinity of lecithin and a polyhydric alcohol due to the interaction with lecithin and a polyhydric alcohol.

The present invention is based on the following unexpected findings: the application of the protamine can improve the intake of the dioscorea villosa root extract by skin cells and the bioavailability, and experiments in fibroblasts and Langerhans cells show that the composition containing the roe protein has higher absorption intensity than the composition without the roe protein, and the data have statistical significance.

In certain embodiments of the present invention, the polyol is present in an amount of 60 to 90% by weight.

In certain embodiments of the present invention, the polyol is selected from any two of glycerol, butanediol, pentanediol, 1,3 propanediol.

In certain embodiments of the present invention, the lecithin is present in an amount of 3.3 to 12% by weight.

In some embodiments of the present invention, the weight percentage of the dioscorea villosa root extract is 3.3-8%.

Preparation method

The composition or the external preparation for skin of the present invention can be prepared by a low-temperature freeze-drying method using a high-pressure homogenizer.

The composition containing the extract of the roots of dioscorea villosa of the present invention can be prepared by the following process. For example, 1a) dispersing a long dioscorea villosa root extract and lecithin in a polyol to form a polyol system; 1b) lyophilizing or homogenizing under high pressure to obtain composition containing extract of Dioscorea villosa root;

in some embodiments of the invention, the resulting composition comprising the extract of the roots of Dioscorea villosa is frozen in a freezer at-80 ℃. Then, freeze-drying was performed with the following settings: sublimation drying is carried out at the temperature of-80 to-40 ℃ and the vacuum degree of 1 to 30Pa for 3 to 21 hours, desorption drying is carried out at the temperature of-10 to +20 ℃ and the vacuum degree of 1 to 30Pa for 1 to 10 hours, and the water content is controlled to be 0 to 6 percent.

In some embodiments of the invention, the resulting composition comprising the extract of the roots of Dioscorea villosa is frozen in a freezer at-80 ℃. Then, spray drying was performed with the following settings: the air inlet temperature is 40-80 ℃, the atomization pressure is 0.2-0.8 MPa, the feeding speed is 100-2000 ml/h, and the water content is controlled to be 0-8%.

In some embodiments of the invention, the resulting composition comprising the extract of dioscorea villosa root is homogenized in a high pressure homogenizer: the circulation is carried out for 3 times at the flow rate of 1200L/h and the pressure of 100 MPa.

The skin external preparation containing the extract of the root of dioscorea villosa according to the present invention can be prepared by the following process. For example,

1a) dispersing a long dioscorea villosa root extract and lecithin in a polyol to form a polyol system; 1b) lyophilizing or homogenizing under high pressure to obtain composition containing extract of Dioscorea villosa root; 1c) mixing the fish egg protein with the composition comprising the extract of the roots of dioscorea villosa prepared in step 1b) to obtain the skin external preparation comprising the fish egg protein.

In some embodiments of the present invention, the obtained skin external composition containing the extract of the root of dioscorea villosa is frozen in a freezer at-80 ℃. Then, freeze-drying was performed with the following settings: sublimation drying is carried out at the temperature of-80 to-40 ℃ and the vacuum degree of 1 to 30Pa for 3 to 21 hours, desorption drying is carried out at the temperature of-10 to +20 ℃ and the vacuum degree of 1 to 30Pa for 1 to 10 hours, and the water content is controlled to be 0 to 6 percent.

In some embodiments of the present invention, the obtained skin external preparation containing the extract of the root of dioscorea villosa is frozen in a freezer at-80 ℃. Then, spray drying was performed with the following settings: the air inlet temperature is 40-80 ℃, the atomization pressure is 0.2-0.8 MPa, the feeding speed is 100-2000 ml/h, and the water content is controlled to be 0-8%.

In some embodiments of the present invention, the obtained external preparation for skin containing the extract of dioscorea villosa root is subjected to a homogenization treatment in a high pressure homogenizer: the circulation is carried out for 3 times at the flow rate of 1200L/h and the pressure of 100 MPa.

Application method

The composition of the present invention comprising an extract of dioscorea villosa root may be topically applied to mammalian skin. For example, the composition comprising the extract of dioscorea villosa root can be applied to skin in need of treatment from up to 2 times per day to as little as 1 time per week (e.g., once per day, once every two days, once per week), and the like.

The external preparation for skin containing the extract of dioscorea villosa root of the present invention may be topically applied to the skin of a mammal. For example, the composition comprising the extract of dioscorea villosa root can be applied to skin in need of treatment from up to 2 times per day to as little as 1 time per week (e.g., once per day, once every two days, once per week), and the like.

The use of the fish egg protein can improve the intake of skin cells to the composition, improve the bioavailability and the permeability, and show that the fish egg protein composition has higher absorption intensity compared with the common composition in skin model experiments, and has statistical significance among data.

The technical aspects of the present invention will be described in detail below with reference to preferred embodiments, but the scope of the present invention is not limited to these embodiments, and the technical aspects of the present invention are intended to be described and not limited. Test methods in which specific conditions are not specified in the following examples are generally carried out under conventional conditions or under conditions recommended by the manufacturer. All percentages and parts are by weight unless otherwise indicated.

The source description of the raw materials in the invention (type and manufacturer, list or text description)

Long mollissima rhizome extract: guangzhou home Biotechnology Ltd

Roe protein (P4020): Sigma-ALORICH Co

The coumarins are all obtained from SIGMA-ALORICH

Lecithin: japanese Kokusan Kogyo

Glycerol, pentanediol, butanediol, 1, 3-propanediol, coumarin, etc., all of analytical purity, Shanghai Allan Biochemical department

Flow cytometer, model FACSCANTO 10C, BD Biosciences

Vacuum freeze dryer model FD-2, Beijing Bo Yi kang laboratory instruments Ltd

Spray drier model QFN-9000Y, Shanghai georgen industries Ltd

The in vitro fibroblast, melanocyte, and Langerhans cell culture method adopts enzyme digestion method, i.e. MEM complete culture solution at 37 deg.C and 5% CO₂Culturing in an incubator.

Example 1:

41.7 g of glycerol and 30 g of pentanediol are respectively weighed and added into a beaker, 3.3 g of lecithin, 8.3 g of the dioscorea villosa root extract and 0.2 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 200 rpm. After mixing was complete, the mixture was heated to 60 ℃ in a water bath, the stirring speed was maintained at 200rpm, and stirring was maintained for 20 minutes. Then, the temperature was lowered by stirring with a disk-type stirring paddle, the stirring speed was maintained at 150rpm, and the stirring was stopped after cooling to 45 ℃. Freeze drying, setting sublimation drying at-80 to-40 ℃, vacuum degree of 1 to 30Pa for 12 hours, setting desorption drying at-10 to +20 ℃, vacuum degree of 1 to 30Pa for 8 hours, controlling water content between 0 and 6 percent, setting sublimation drying at-80 to-40 ℃, vacuum degree of 1 to 30Pa for 12 hours, setting desorption drying at-10 to +20 ℃, vacuum degree of 1 to 30Pa for 10 hours, controlling water content between 0 and 6 percent, and obtaining white loose porous sponge-shaped amorphous powder. Adding 16.7 g of roe protein, grinding, and sieving by a 300-mesh sieve to obtain a white composition for later use;

example 2:

33.3 g of glycerol, 26.7 g of butanediol and 16.7 g of 1, 3-propanediol are respectively weighed and added into a beaker, 3.3 g of lecithin, 3.3 g of long-hair dioscorea root extract and 0.04 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 220 rpm. After mixing was complete, the mixture was heated to 70 ℃ with a water bath, the stirring speed was maintained at 220rpm, and stirring was maintained for 12 minutes. Then, the temperature was lowered by stirring with a disk-type stirring paddle, the stirring speed was maintained at 120rpm, and the stirring was stopped after cooling to 40 ℃. Freeze drying, setting sublimation drying at-80 to-40 ℃, vacuum degree of 1 to 30Pa, time of 17 hours, setting desorption drying at-10 to +20 ℃, vacuum degree of 1 to 30Pa, time of 10 hours, controlling water content between 0 and 6 percent, setting sublimation drying at-80 to-40 ℃, vacuum degree of 1 to 30Pa, time of 5 hours, setting desorption drying at-10 to +20 ℃, vacuum degree of 1 to 30Pa, time of 5 hours, controlling water content between 0 and 6 percent, and obtaining white loose porous sponge-shaped amorphous powder. Adding 16.7 g of roe protein, grinding, and sieving by a 300-mesh sieve to obtain a white composition for later use;

example 3:

33.3 g of glycerin and 33.3 g of butanediol are respectively weighed and added into a beaker, 6.7 g of lecithin, 10 g of the dioscorea villosa root extract and 0.05 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 280 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 280rpm, and stirring was maintained for 15 minutes. And then stirring and cooling by using a disc type stirring paddle, keeping the stirring speed at 200rpm, stopping stirring after cooling to 45 ℃, freeze-drying, setting sublimation drying at the temperature of-80 to-40 ℃, the vacuum degree at 1 to 30Pa, keeping the time for 15 hours, setting desorption drying at the temperature of-10 to +20 ℃, setting the vacuum degree at 1 to 30Pa, keeping the time for 6 hours, controlling the water content at 0 to 6 percent, setting sublimation drying at the temperature of-80 to-40 ℃, setting the vacuum degree at 1 to 30Pa, keeping the time for 18 hours, setting desorption drying at the temperature of-10 to +20 ℃, keeping the vacuum degree at 1 to 30Pa, keeping the time for 3 hours, and keeping the water content at 0 to 6 percent to obtain white loose porous spongy amorphous powder. Adding 16.7 g of roe protein, grinding, and sieving by a 300-mesh sieve to obtain a white composition for later use;

example 4:

16.7 g of glycerin and 29.1 g of butanediol are respectively weighed and added into a beaker, and then 20.8 g of lecithin, 16.7 g of the dioscorea villosa root extract and 0.1 g of coumarin are weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 300 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 300rpm, and stirring was maintained for 5 minutes. Then, the temperature was lowered by stirring with a disk-type stirring paddle, the stirring speed was maintained at 200rpm, and the stirring was stopped after cooling to 45 ℃. Freeze drying, setting sublimation drying at-80 to-40 ℃, the vacuum degree of 1 to 30Pa, the time of 20 hours, setting desorption drying at-10 to +20 ℃, the vacuum degree of 1 to 30Pa, the time of 2 hours, and controlling the water content between 0 and 6 percent, setting sublimation drying at-80 to-40 ℃, the vacuum degree of 1 to 30Pa, the time of 10 hours, setting desorption drying at-10 to +20 ℃, the vacuum degree of 1 to 30Pa, the time of 7 hours, and controlling the water content between 0 and 6 percent, and obtaining the white loose porous sponge-shaped amorphous powder. Adding 16.7 g of roe protein, grinding, and sieving by a 300-mesh sieve to obtain a white composition for later use;

example 5:

55 g of glycerin and 16.7 g of butanediol are respectively weighed and added into a beaker, 5 g of lecithin, 6.6 g of the long-hair dioscorea root extract and 0.08 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 350 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 350rpm, and stirring was maintained for 6 minutes. Then, the temperature was lowered by stirring with a disk-type stirring paddle, the stirring speed was maintained at 150rpm, and the stirring was stopped after cooling to 40 ℃. Freeze drying, setting sublimation drying at-80 to-40 ℃, vacuum degree of 1 to 30Pa, time of 5 hours, setting desorption drying at-10 to +20 ℃, vacuum degree of 1 to 30Pa, time of 9 hours, controlling water content between 0 and 6 percent, setting sublimation drying at-80 to-40 ℃, vacuum degree of 1 to 30Pa, time of 11 hours, setting desorption drying at-10 to +20 ℃, vacuum degree of 1 to 30Pa, time of 4 hours, controlling water content between 0 and 6 percent, and obtaining white loose porous sponge-shaped amorphous powder. Adding 16.7 g of roe protein, grinding, and sieving by a 300-mesh sieve to obtain a white composition for later use;

example 6:

25 g of glycerin and 29.2 g of butanediol are respectively weighed and added into a beaker, and 16.6 g of lecithin, 12.5 g of the dioscorea villosa root extract and 0.15 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 400 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 400rpm, and stirring was maintained for 15 minutes. Then, the temperature was lowered by stirring with a disk-type stirring paddle, the stirring speed was maintained at 200rpm, and the stirring was stopped after cooling to 40 ℃. Freeze drying, setting sublimation drying at-80 to-40 ℃, the vacuum degree of 1 to 30Pa, the time of 21 hours, setting desorption drying at-10 to +20 ℃, the vacuum degree of 1 to 30Pa, the time of 1 hour, and the water content of 0 to 6 percent, setting sublimation drying at-80 to-40 ℃, the vacuum degree of 1 to 30Pa, the time of 8 hours, setting desorption drying at-10 to +20 ℃, the vacuum degree of 1 to 30Pa, and the water content of 6 hours, and controlling the water content of 0 to 6 percent, thereby obtaining the white loose porous sponge-shaped amorphous powder. Adding 16.7 g of roe protein, grinding, and sieving by a 300-mesh sieve to obtain a white composition for later use;

example 7:

33.3 g of glycerol and 26.7 g of pentanediol are respectively weighed and added into a beaker, 6.7 g of lecithin, 16.6 g of the dioscorea villosa root extract and 0.2 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 400 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 400rpm, and stirring was maintained for 15 minutes. After stirring, homogenizing the composition by using a high-pressure homogenizer at a flow rate of 1200L/h and a pressure of 100MPa for 3 times, circulating for 3 times to obtain milky smooth and fine emulsion, adding 16.7 g of roe protein, and uniformly stirring to obtain the composition for later use;

example 8:

33.3 g of glycerol, 26.7 g of butanediol and 16.7 g of 1, 3-propanediol are respectively weighed and added into a beaker, 3.3 g of lecithin, 3.3 g of long-hair dioscorea root extract and 0.04 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 400 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 400rpm, and stirring was maintained for 10 minutes. After stirring, homogenizing the composition by using a high-pressure homogenizer at a flow rate of 1200L/h and a pressure of 100MPa for 3 times, circulating for 3 times to obtain milky smooth and fine emulsion, adding 16.7 g of roe protein, and uniformly stirring to obtain the composition for later use;

example 9:

33.3 g of glycerin and 42.5 g of butanediol are respectively weighed and added into a beaker, and 3.3 g of lecithin, 4.2 g of the dioscorea villosa root extract and 0.05 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 400 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 400rpm, and stirring was maintained for 10 minutes. After stirring, homogenizing the composition by using a high-pressure homogenizer at a flow rate of 1200L/h and a pressure of 100MPa for 3 times, circulating for 3 times to obtain milky smooth and fine emulsion, adding 16.7 g of roe protein, and uniformly stirring to obtain the composition for later use;

example 10:

16.7 g of glycerol and 33.3 g of butanediol are respectively weighed and added into a beaker, and 25 g of lecithin, 8.3 g of the dioscorea villosa root extract and 0.1 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 400 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 400rpm, and stirring was maintained for 10 minutes. After stirring, homogenizing the composition by using a high-pressure homogenizer at a flow rate of 1200L/h and a pressure of 100MPa for 3 times, circulating for 3 times to obtain milky smooth and fine emulsion, adding 16.7 g of roe protein, and uniformly stirring to obtain the composition for later use;

example 11:

55 g of glycerin and 16.7 g of butanediol are respectively weighed and added into a beaker, 5 g of lecithin, 6.6 g of the extract of the long-hair dioscorea root and 0.08 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 400 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 400rpm, and stirring was maintained for 10 minutes. After stirring, homogenizing the composition by using a high-pressure homogenizer at a flow rate of 1200L/h and a pressure of 100MPa for 3 times, circulating for 3 times to obtain milky smooth and fine emulsion, adding 16.7 g of roe protein, and uniformly stirring to obtain the composition for later use;

example 12:

25 g of glycerin and 29.2 g of butanediol are respectively weighed and added into a beaker, and 16.6 g of lecithin, 12.5 g of the dioscorea villosa root extract and 0.15 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 400 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 400rpm, and stirring was maintained for 10 minutes. After stirring, homogenizing the composition by using a high-pressure homogenizer at a flow rate of 1200L/h and a pressure of 100MPa for 3 times, circulating for 3 times to obtain milky smooth and fine emulsion, adding 16.7 g of roe protein, and uniformly stirring to obtain the composition for later use;

example 13:

41.7 g of glycerol and 30 g of pentanediol are respectively weighed and added into a beaker, 3.3 g of lecithin, 8.3 g of the dioscorea villosa root extract and 0.2 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 200 rpm. After mixing was complete, the mixture was heated to 60 ℃ in a water bath, the stirring speed was maintained at 200rpm, and stirring was maintained for 20 minutes. Then, the temperature was lowered by stirring with a disk-type stirring paddle, the stirring speed was maintained at 150rpm, and the stirring was stopped after cooling to 45 ℃. Freeze drying, setting sublimation drying at-80 to-40 ℃, vacuum degree of 1 to 30Pa for 12 hours, setting desorption drying at-10 to +20 ℃, vacuum degree of 1 to 30Pa for 8 hours, controlling water content between 0 and 6 percent, setting sublimation drying at-80 to-40 ℃, vacuum degree of 1 to 30Pa for 12 hours, setting desorption drying at-10 to +20 ℃, vacuum degree of 1 to 30Pa for 10 hours, controlling water content between 0 and 6 percent, and obtaining white loose porous sponge-shaped amorphous powder. Grinding, and sieving with 300 mesh sieve to obtain white composition;

example 14:

33.3 g of glycerol, 26.7 g of butanediol and 16.7 g of 1, 3-propanediol are respectively weighed and added into a beaker, 3.3 g of lecithin, 3.3 g of long-hair dioscorea root extract and 0.04 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 220 rpm. After mixing was complete, the mixture was heated to 70 ℃ with a water bath, the stirring speed was maintained at 220rpm, and stirring was maintained for 12 minutes. Then, the temperature was lowered by stirring with a disk-type stirring paddle, the stirring speed was maintained at 120rpm, and the stirring was stopped after cooling to 40 ℃. Freeze drying, setting sublimation drying at-80 to-40 ℃, vacuum degree of 1 to 30Pa, time of 17 hours, setting desorption drying at-10 to +20 ℃, vacuum degree of 1 to 30Pa, time of 10 hours, controlling water content between 0 and 6 percent, setting sublimation drying at-80 to-40 ℃, vacuum degree of 1 to 30Pa, time of 5 hours, setting desorption drying at-10 to +20 ℃, vacuum degree of 1 to 30Pa, time of 5 hours, controlling water content between 0 and 6 percent, and obtaining white loose porous sponge-shaped amorphous powder. Grinding, and sieving with 300 mesh sieve to obtain white composition;

example 15:

33.3 g of glycerin and 33.3 g of butanediol are respectively weighed and added into a beaker, 6.7 g of lecithin, 10 g of the dioscorea villosa root extract and 0.05 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 280 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 280rpm, and stirring was maintained for 15 minutes. And then stirring and cooling by using a disc type stirring paddle, keeping the stirring speed at 200rpm, stopping stirring after cooling to 45 ℃, freeze-drying, setting sublimation drying at the temperature of-80 to-40 ℃, the vacuum degree at 1 to 30Pa, keeping the time for 15 hours, setting desorption drying at the temperature of-10 to +20 ℃, setting the vacuum degree at 1 to 30Pa, keeping the time for 6 hours, controlling the water content at 0 to 6 percent, setting sublimation drying at the temperature of-80 to-40 ℃, setting the vacuum degree at 1 to 30Pa, keeping the time for 18 hours, setting desorption drying at the temperature of-10 to +20 ℃, keeping the vacuum degree at 1 to 30Pa, keeping the time for 3 hours, and keeping the water content at 0 to 6 percent to obtain white loose porous spongy amorphous powder. Grinding, and sieving with 300 mesh sieve to obtain white composition;

example 16:

16.7 g of glycerin and 29.1 g of butanediol are respectively weighed and added into a beaker, and then 20.8 g of lecithin, 16.7 g of the dioscorea villosa root extract and 0.1 g of coumarin are weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 300 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 300rpm, and stirring was maintained for 5 minutes. Then, the temperature was lowered by stirring with a disk-type stirring paddle, the stirring speed was maintained at 200rpm, and the stirring was stopped after cooling to 45 ℃. Freeze drying, setting sublimation drying at-80 to-40 ℃, the vacuum degree of 1 to 30Pa, the time of 20 hours, setting desorption drying at-10 to +20 ℃, the vacuum degree of 1 to 30Pa, the time of 2 hours, and controlling the water content between 0 and 6 percent, setting sublimation drying at-80 to-40 ℃, the vacuum degree of 1 to 30Pa, the time of 10 hours, setting desorption drying at-10 to +20 ℃, the vacuum degree of 1 to 30Pa, the time of 7 hours, and controlling the water content between 0 and 6 percent, and obtaining the white loose porous sponge-shaped amorphous powder. Grinding, and sieving with 300 mesh sieve to obtain white composition;

example 17:

55 g of glycerin and 16.7 g of butanediol are respectively weighed and added into a beaker, 5 g of lecithin, 6.6 g of the long-hair dioscorea root extract and 0.08 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 350 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 350rpm, and stirring was maintained for 6 minutes. Then, the temperature was lowered by stirring with a disk-type stirring paddle, the stirring speed was maintained at 150rpm, and the stirring was stopped after cooling to 40 ℃. Freeze drying, setting sublimation drying at-80 to-40 ℃, vacuum degree of 1 to 30Pa, time of 5 hours, setting desorption drying at-10 to +20 ℃, vacuum degree of 1 to 30Pa, time of 9 hours, controlling water content between 0 and 6 percent, setting sublimation drying at-80 to-40 ℃, vacuum degree of 1 to 30Pa, time of 11 hours, setting desorption drying at-10 to +20 ℃, vacuum degree of 1 to 30Pa, time of 4 hours, controlling water content between 0 and 6 percent, and obtaining white loose porous sponge-shaped amorphous powder. Grinding, and sieving with 300 mesh sieve to obtain white composition;

example 18:

25 g of glycerin and 29.2 g of butanediol are respectively weighed and added into a beaker, and 16.6 g of lecithin, 12.5 g of the dioscorea villosa root extract and 0.15 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 400 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 400rpm, and stirring was maintained for 15 minutes. Then, the temperature was lowered by stirring with a disk-type stirring paddle, the stirring speed was maintained at 200rpm, and the stirring was stopped after cooling to 40 ℃. Freeze drying, setting sublimation drying at-80 to-40 ℃, the vacuum degree of 1 to 30Pa, the time of 21 hours, setting desorption drying at-10 to +20 ℃, the vacuum degree of 1 to 30Pa, the time of 1 hour, and the water content of 0 to 6 percent, setting sublimation drying at-80 to-40 ℃, the vacuum degree of 1 to 30Pa, the time of 8 hours, setting desorption drying at-10 to +20 ℃, the vacuum degree of 1 to 30Pa, and the water content of 6 hours, and controlling the water content of 0 to 6 percent, thereby obtaining the white loose porous sponge-shaped amorphous powder. Grinding, and sieving with 300 mesh sieve to obtain white composition;

example 19:

33.3 g of glycerol and 26.7 g of pentanediol are respectively weighed and added into a beaker, and 6.7 g of lecithin, 16.7 g of the dioscorea villosa root extract and 0.2 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 400 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 400rpm, and stirring was maintained for 15 minutes. After stirring, homogenizing the composition by using a high-pressure homogenizer, circulating for 3 times at a flow rate of 1200L/h and a pressure of 100MPa to obtain milky smooth and fine emulsion, and uniformly stirring to obtain the composition for later use;

example 20:

33.3 g of glycerol, 26.7 g of butanediol and 16.7 g of 1, 3-propanediol are respectively weighed and added into a beaker, 3.3 g of lecithin, 3.3 g of long-hair dioscorea root extract and 0.04 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 400 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 400rpm, and stirring was maintained for 10 minutes. After stirring, homogenizing the composition by using a high-pressure homogenizer, circulating for 3 times at a flow rate of 1200L/h and a pressure of 100MPa to obtain milky smooth and fine emulsion, and uniformly stirring to obtain the composition for later use;

example 21:

33.3 g of glycerin and 42.5 g of butanediol are respectively weighed and added into a beaker, and 3.3 g of lecithin, 4.2 g of the dioscorea villosa root extract and 0.05 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 400 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 400rpm, and stirring was maintained for 10 minutes. After stirring, homogenizing the composition by using a high-pressure homogenizer, circulating for 3 times at a flow rate of 1200L/h and a pressure of 100MPa to obtain milky smooth and fine emulsion, and uniformly stirring to obtain the composition for later use;

example 22:

16.7 g of glycerol and 33.3 g of butanediol are respectively weighed and added into a beaker, and 25 g of lecithin, 8.3 g of the dioscorea villosa root extract and 0.1 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 400 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 400rpm, and stirring was maintained for 10 minutes. After stirring, homogenizing the composition by using a high-pressure homogenizer, circulating for 3 times at a flow rate of 1200L/h and a pressure of 100MPa to obtain milky smooth and fine emulsion, and uniformly stirring to obtain the composition for later use;

example 23:

55 g of glycerin and 16.7 g of butanediol are respectively weighed and added into a beaker, 5 g of lecithin, 6.6 g of the extract of the long-hair dioscorea root and 0.08 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 400 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 400rpm, and stirring was maintained for 10 minutes. After stirring, homogenizing the composition by using a high-pressure homogenizer, circulating for 3 times at a flow rate of 1200L/h and a pressure of 100MPa to obtain milky smooth and fine emulsion, and uniformly stirring to obtain the composition for later use;

example 24:

25 g of glycerin and 29.2 g of butanediol are respectively weighed and added into a beaker, and 16.6 g of lecithin, 12.5 g of the dioscorea villosa root extract and 0.15 g of coumarin are further weighed and added into the same beaker, and are uniformly mixed by a disc type stirring paddle, and the stirring speed is kept at 400 rpm. After mixing was complete, the mixture was heated to 75 ℃ in a water bath, the stirring speed was maintained at 400rpm, and stirring was maintained for 10 minutes. After stirring, homogenizing the composition by using a high-pressure homogenizer, circulating for 3 times at a flow rate of 1200L/h and a pressure of 100MPa to obtain milky smooth and fine emulsion, and uniformly stirring to obtain the composition for later use;

test example 1 stability of the composition

The samples were filled into 100ml glass vials and stored in thermostats at different temperatures. The appearance and structural change of the samples were observed at different times.

The laboratory high-temperature and low-temperature storage stability test can be carried out by adopting the following method: (1) placing the mixture in a constant temperature incubator at 48 ℃ for 90 days, and observing after the temperature is recovered, and (2) placing the mixture in a constant temperature incubator at 25 ℃ for 90 days, and observing the stability and the stability after the temperature is recovered; (3) the stability was observed after storage at-18 ℃ for 90 days and return to room temperature.

And if the sample has no obvious oil-water separation, no obvious mildew and bacteria growing, no obvious odor and color change, the stability inspection is passed. The results are shown in Table 1.

Table 1 stability results for compositions of examples 1-24

In examples 1-24, examples 4, 6,10,12,16,18,22 failed the immediate, one month and three month high temperature stability test, and the composition delaminated, flocculated, coarsened and yellowed during the test.

Test example 2 evaluation of safety

Examples 1-24 were formulated as 10% aqueous solutions, and then skin irritation experiments were performed on the 10% aqueous solutions.

The experimental animals used are adult, healthy, non-skin damaged, normal white New Zealand rabbits, weighing 2.3-2.6kg, female and male, but female animals should be infertile and have not been born. At least 4 animals were used. The experimental animals are raised in single cages, and the animals are adapted to the environment of the experimental animal room for at least 3 days before the experiment, and the temperature is controlled to be 19-26 ℃. The experimental animals and the experimental animal rooms conform to the corresponding national regulations. The conventional feed is selected, and drinking water is not limited. The hair on both sides of the spine of the experimental animal is cut off about 24 hours before the experiment, the epidermis cannot be damaged, and the hair removing range is about 3cm multiplied by 3cm on the left and the right respectively. Approximately 0.5mL (g) of the test article is applied directly to one side of the skin and then covered with two layers of gauze (2.5 cm. times.2.5 cm) and a layer of cellophane or the like, and secured with a non-irritating adhesive tape and bandage. The other side of the skin was treated with distilled water as a control. The application time is 4h by adopting a sealing test. After the test is finished, the residual test substance is removed by warm water or a non-irritant solvent. Skin reactions of the smeared parts are observed for 1, 24, 48 and 72h after the removal of the test substances, skin reaction scores are carried out according to the table 1, comprehensive evaluation is carried out according to the average value of the tested animal integrals, and the skin irritation intensity is judged according to the highest integral average value of each observation time point of 24, 48 and 72 h.

The average integral per animal per day was calculated according to the following formula, and the skin irritation intensity was judged in tables 2 and 3.

Average score per animal per day ═ (Σ integration of erythema and edema/number of animals tested)/14

TABLE 2 skin irritation response score

TABLE 3 skin irritation Strength grading

[ evaluation results ]

TABLE 4 safety evaluation results

Table 4 the compositions of examples 1-3, 5, 7-9, 11, 13-15, 17, 19-21, and 23, which were 10% aqueous solutions, all had erythema and redness scores of less than 0.5 points and were judged to be non-irritating, and the compositions prepared in these examples were safe. On the other hand, the 10% aqueous solution of the composition of examples 4, 6,10,12,16,18,22,24 had erythema and redness scores between 0.5 and 2.0, and showed marked erythema and redness with mild irritation, and failed to be judged.

Test example 3

Fibroblasts were cultured in a conventional manner, plated (6-well plates), and the composition with added egg protein of examples 1-12 (blank) and the composition with added egg protein of examples 13-24 were used. Adding a composition with a fluorescent probe (coumarin) -culture medium solution into the blank group, adding the roe protein with the fluorescent probe (coumarin) into the culture medium solution, wherein the concentration of the roe protein is 25ug/ml, incubating the cells for 4h, and treating the cells: after incubation, removing the culture medium, adding 300ul of pancreatin into each well, digesting for 2min, adding the culture medium to stop digestion, beating for three times, centrifuging (1200r/min for 3min), discarding the supernatant, taking the precipitate, adding 1ml of PBS to the precipitate, beating, centrifuging (1200r/min for 3min), discarding the supernatant, adding 0.3ml of PBS to the precipitate, beating, measuring the cell uptake by using a cell flow meter, and detecting by using a computer. Data differences were statistically significant (P < 0.01).

The fluorescent probe is added into the composition in the preparation process as a marker;

the median FL1H (coumarin-corresponding detection channel) in the flow cytometer data was statistically averaged and the results are shown in table 5 for the composition of each of the compositions prepared in examples 1-24 and the results in the fibroblast experiments.

TABLE 5

Examples	Fibroblast uptake/FL 1H	Examples	Fibroblast uptake/FL 1H
				1	89395	13	54181
2	89776	14	49776
				3	85432	15	37651
4	87690	16	41996
				5	79651	17	50872
6	77895	18	33497
				7	80584	19	45809
8	78951	20	44361
				9	77503	21	48952
10	82369	22	53490
				11	81207	23	49810
12	78699	24	47658

The compositions prepared in examples 1 to 6 and examples 13 to 18 were lyophilized compositions prepared by the same lyophilization process, with the difference that examples 1 to 6 were lyophilized compositions to which fish egg protein was added, and examples 13 to 18 were lyophilized compositions to which fish egg protein was not added. Specifically, the enhancement rate was 65% for example 1 compared to the median mean FL1H of the lyophilized composition of example 13; example 2 the boost rate was 80.4% compared to the median mean FL1H of the lyophilized composition of example 14; example 3 the boost rate was 126.9% compared to the median mean FL1H of the lyophilized composition of example 15; example 4 the boost rate was 108.8% compared to the median mean FL1H of the lyophilized composition of example 16; example 5 the boost rate was 56.6% compared to the median mean FL1H of the lyophilized composition of example 17; example 6 the boost rate was 132.5% compared to the median mean FL1H of the lyophilized composition of example 18; the average lift was 95%. It can be seen that the lyophilized compositions of examples 1-6 can be more easily absorbed by fibroblasts than the lyophilized compositions of examples 13-18 due to the addition of the fish egg protein.

The compositions prepared in examples 7-12 and examples 19-24 were prepared by the same high pressure homogenization procedure, with the difference that examples 7-12 were compositions with added egg protein, while examples 19-24 were compositions without added egg protein. Specifically, the boost rate of example 7 was 75.9% compared to the median mean FL1H of the composition of example 19; example 8 the boost rate was 78.0% compared to the median mean FL1H for the composition of example 20; example 9 the boost rate was 58.3% compared to the median mean FL1H for the composition of example 21; example 10 had a boost rate of less than 54.0% compared to the median mean FL1H of the composition of example 22; example 11 had a 63% boost compared to the median mean FL1H for the composition of example 23; example 12 the boost rate was 65.1% compared to the median mean FL1H of the lyophilized composition of example 24; the average lift was 65.7%. It can be seen that the lyophilized compositions of examples 7-12 can be more easily absorbed by fibroblasts than the compositions of examples 19-24 due to the addition of the fish egg protein.

In summary, it can be seen that the average increasing rate of FL1H median mean value by the freeze-drying process and the superposition of roe protein is 95%, which is much higher than the average increasing rate of FL1H median mean value by the homogenization process and the superposition of roe protein 65.7%.

Test example 4

Langerhans cells were grown by conventional methods, plated (6-well plates), with examples 13-24 as no egg protein added (blank) and examples 1-12 as egg protein added. Adding the long-hair dioscorea root extract composition with a fluorescent probe (coumarin) -culture medium solution (the concentration of the long-hair dioscorea root extract is 50ug/ml) into the blank group, adding the long-hair dioscorea root extract composition with the fluorescent probe (coumarin) and the roe protein-culture medium solution (the concentration of the long-hair dioscorea root extract is 50ug/ml and the concentration of the roe protein is 25ug/ml) into the roe protein group, incubating the cells for 4h, and treating the cells: after incubation, removing the culture medium, adding 300ul of pancreatin into each well, digesting for 2min, adding the culture medium to stop digestion, beating for three times, centrifuging (1200r/min for 3min), discarding the supernatant, taking the precipitate, adding 1ml of PBS to the precipitate, beating, centrifuging (1200r/min for 3min), discarding the supernatant, adding 0.3ml of PBS to the precipitate, beating, measuring the cell uptake by using a cell flow meter, and detecting by using a computer. Data differences were statistically significant (P < 0.01).

TABLE 6

Examples	Langerhans cell uptake results/FL 1H	Examples	Langerhans cell uptake results/FL 1H
				1	16207	13	7936
2	17346	14	7659
				3	12390	15	6651
4	13507	16	6932
				5	15711	17	6834
6	16589	18	6654
				7	14572	19	5912
8	16604	20	7730
				9	15488	21	7129
10	14507	22	7323
				11	15612	23	6873
12	14382	24	6751

The compositions prepared in examples 1 to 6 and examples 13 to 18 were lyophilized compositions prepared by the same lyophilization process, with the difference that examples 1 to 6 were lyophilized compositions to which fish egg protein was added, and examples 13 to 18 were lyophilized compositions to which fish egg protein was not added. Specifically, the enhancement rate of example 1 was 104.2% compared to the median mean FL1H of the lyophilized composition of example 13; example 2 the boost rate was 126.5% compared to the median mean FL1H of the lyophilized composition of example 14; example 3 the boost rate was 86.3% compared to the median mean FL1H of the lyophilized composition of example 15; example 4 the boost rate was 94.8% compared to the median mean FL1H of the lyophilized composition of example 16; example 5 the boost rate was 129.9% compared to the median mean FL1H of the lyophilized composition of example 17; example 6 enhancement rate was 149.3% compared to the median mean FL1H of the lyophilized composition of example 18; the average lift was 115.2%. It can be seen that the lyophilized compositions of examples 1-6 can be more easily absorbed by langerhans cells than the lyophilized compositions of examples 13-18 due to the addition of the fish egg protein.

The compositions prepared in examples 7-12 and examples 19-24 were prepared by the same high pressure homogenization procedure, with the difference that examples 7-12 were compositions with added egg protein, while examples 19-24 were compositions without added egg protein. Specifically, the boost rate was 146.5% for example 7 compared to the median mean FL1H for the composition of example 19; example 8 the boost rate was 114.8% compared to the median mean FL1H for the composition of example 20; example 9 the boost rate compared to the median mean FL1H for the composition of example 21 was 117.3%; example 10 had a boost rate of less than 98.1% compared to the median mean FL1H of the composition of example 22; example 11 the boost rate was 127.1% compared to the median mean FL1H for the composition of example 23; example 12 the boost rate was 113% compared to the median mean FL1H of the lyophilized composition of example 24; the average lift was 119.5%. It can be seen that the lyophilized compositions of examples 7-12 can be more easily absorbed by langerhans cells than the compositions of examples 19-24 due to the addition of the fish egg protein.

In summary, it can be seen that the average increasing rate of FL1H median mean value by the freeze-drying process and the superposition of roe protein is 115.2%, which is comparable to the average increasing rate of FL1H median mean value by the homogenization process and the superposition of roe protein is 119.5%.

Test example 5: electron microscopy characterization

The composition prepared in example 1 was used as a subject to be examined for microscopic morphology characterization (test image is an effect of 2.5 ten thousand times magnification of an actual sample) using an apparatus of an electron microscope (model: AMT Camera), and the analysis test results are shown in FIG. 1 below.

As can be seen from the electron microscopic scanning results, the composition of example 1 comprises dioscorea villosa root extract, lecithin, polyol and fish egg protein, and has an "onion" -type layer-by-layer microcapsule structure.

The compositions prepared in examples, which were successful in stability and safety, were used for the preparation of external preparations for skin. The skin external preparation is preferably a cosmetic, such as a lotion, essence, cream, etc. The weight percentage of the composition in the skin external preparation is 0.001-50% (w/w). The preferred weight percentage is 1% to 20% (w/w). More preferably 2% to 10% (w/w). The most preferred weight percentage is 3% to 8% (w/w).

The following are examples of specific applications of the compositions in skin external preparations, and formulations and methods for preparing these dosage forms. In the tables, "-" indicates no addition.

Application example 1: preparation of face cream

The raw materials and contents are shown in Table 7

TABLE 7

Application example 2: preparation of the emulsion

The raw materials and contents are shown in Table 8.

TABLE 8

Application example 3: preparation of jelly

The raw materials and contents are shown in Table 9.

TABLE 9

Application example 4: preparation of astringent

The raw materials and contents are shown in Table 10.

Watch 10

Application example 5: preparation of essence

The raw materials and contents are shown in Table 11.

TABLE 11

Application example 6: preparation of facial mask

The raw materials and contents are shown in Table 12.

TABLE 12

Application example 7: preparation of eye cream

The raw materials and contents are shown in Table 13.

Watch 13

Application example 8: preparation of shower gel

The raw materials and contents are shown in Table 14.

TABLE 14

Example 9: preparation of facial cleanser

The raw materials and contents are shown in Table 15.

Watch 15

Examples 1 to 6 and examples 13 to 18 in the present invention can also be used alone as a low temperature freeze-dried product.

The cosmetic prepared by the composition of the invention achieves the using effect equivalent to or better than that of the commercial product. The applicant states that the present invention is illustrated in detail by the above examples, but the present invention is not limited to the above detailed methods, i.e. it is not meant that the present invention must rely on the above detailed methods for its implementation. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.

Claims

1. A composition comprising an extract of dioscorea hispida root, comprising: polyhydric alcohol, lecithin and extract of long hair yam root.

2. The composition according to claim 1, wherein the polyol is present in an amount of 60 to 90% by mass.

3. The composition of any of claims 1-2, wherein the polyol is selected from any two of glycerol, butylene glycol, pentylene glycol, 1,3 propanediol.

4. The composition according to claim 1, wherein the lecithin is 1-12% by mass.

5. The composition according to claim 1, wherein the weight percentage of the long dioscorea villosa root extract is 0.1-8%.

6. A process for the preparation of a composition according to claims 1-5, said process comprising the steps of:

7. the method of claim 6, wherein the freeze-drying is configured to: sublimation drying is carried out at the temperature of-80 to-40 ℃ and the vacuum degree of 1 to 30Pa for 3 to 21 hours, desorption drying is carried out at the temperature of-10 to +20 ℃ and the vacuum degree of 1 to 30Pa for 1 to 10 hours, and the water content is controlled to be 0 to 6 percent.

8. The method of claim 6, wherein the high pressure homogenization is configured to: the circulation is carried out for 3 times at the flow rate of 1200L/h and the pressure of 100 MPa.

9. The external preparation for skin comprising the composition according to any one of claims 1 to 5, wherein the external preparation for skin further comprises roe protein.

10. The external preparation for skin as claimed in claim 9, wherein the fish egg protein is selected from the group consisting of: salmon protamine, trout protamine, herring protamine.

11. The method for preparing the external preparation for skin according to claims 9 to 10, comprising the steps of:

12. The method of claim 11, wherein the freeze-drying is configured to: sublimation drying is carried out at the temperature of-80 to-40 ℃ and the vacuum degree of 1 to 30Pa for 3 to 21 hours, desorption drying is carried out at the temperature of-10 to +20 ℃ and the vacuum degree of 1 to 30Pa for 1 to 10 hours, and the water content is controlled to be 0 to 6 percent.

13. The method of claim 11, wherein the high pressure homogenization is configured to: the circulation is carried out for 3 times at the flow rate of 1200L/h and the pressure of 100 MPa.

14. Use of a composition according to claim 1 in cosmetics.

15. The use of the external preparation for skin according to claim 9 in cosmetics.