CN113876633A

CN113876633A - Natural vegetable fat compound formed by human body sebum lipid imitation and preparation method and application thereof

Info

Publication number: CN113876633A
Application number: CN202111220129.XA
Authority: CN
Inventors: 吕韵; 丁德文; 陈强
Original assignee: Hangzhou Lanjiang Cosmetics Co ltd
Current assignee: Hangzhou Lanjiang Cosmetics Co ltd
Priority date: 2021-10-20
Filing date: 2021-10-20
Publication date: 2022-01-04
Anticipated expiration: 2041-10-20
Also published as: CN113876633B

Abstract

A natural vegetable fat compound formed by imitating sebum lipid of human body, a preparation method and application thereof, which belong to the technical field of cosmetics. Comprises the following components in parts by weight: 50-60 parts of vegetable triglyceride, 20-30 parts of vegetable wax ester, 4-6 parts of sterols, 11-13 parts of squalene and 0-0.8 part of tocopherol. The natural vegetable oil-fat compound imitating the formation of human sebum lipid and the preparation method and the application thereof construct a stable, mild and non-irritant natural vegetable oil-fat composition, simulate the formation of human sebum and the proportion of fatty acid, thereby obtaining a cosmetic beautifying oil-fat component with high biocompatibility and high bioavailability, being beneficial to enhancing the oxidation protection power of skin, softening and firming skin, maintaining the stable state of skin and achieving the effect of fading fine wrinkles.

Description

Natural vegetable fat compound formed by human body sebum lipid imitation and preparation method and application thereof

Technical Field

The invention belongs to the technical field of cosmetics, and particularly relates to a natural vegetable fat compound formed by human body sebum lipid imitation, a preparation method and application thereof.

Background

With the age, sebum is gradually degraded and the metabolic capability of the human body is reduced, so that the barrier function of the skin is reduced, subcutaneous collagen is lost, and the skin of the human body is gradually aged. The skin barrier is a physical barrier which is formed by a sebum membrane, keratin, lipid, a sandwich structure, a brick wall structure, dermal mucopolysaccharide, mucopolysaccharide and the like, resists the entrance of external harmful substances, irritants and sunlight, and has the effects of moisturizing, regulating and resisting inflammation. The damage of skin physical barrier can cause skin dryness, skin aging, pigmentation atopic dermatitis, eczema, psoriasis, ichthyosis, solar dermatitis, skin sensitivity, irritant dermatitis, hormone dependent dermatitis, seborrheic diseases such as acne, rosacea, and seborrheic dermatitis. At present, a variety of skin barrier function repairing cosmetics are various, inflammation of damaged skin is reduced by enhancing the moisture retention of the skin surface layer or adding a soothing component, and the effects of barrier function damage and sebum loss cannot be fundamentally solved.

Disclosure of Invention

Aiming at the problems in the prior art, the invention aims to design and provide a natural vegetable oil compound formed by simulating human body sebum lipid and a preparation method and a technical scheme of application thereof, the requirement of different skin using crowds on lipid supplementation is met through the bionic combination of the human body sebum, and the oil compound has strong compatibility with human skin, is beneficial to enhancing the oxidation protection power of the skin, softening and firming the skin, maintaining the stable state of the skin and achieving the effect of fading fine wrinkles.

The natural vegetable fat compound formed by the simulated human sebum lipid is characterized by comprising the following components in parts by weight: 50-60 parts of vegetable triglyceride, 20-30 parts of vegetable wax ester, 4-6 parts of sterols, 11-13 parts of squalene and 0-0.8 part of tocopherols;

the vegetable triglyceride consists of the following raw materials in parts by weight: 13-24 parts of meadowfoam seed oil, 25-40 parts of macadamia nut oil, 0.18-2.3 parts of camelina sativa seed oil, 0.2-2.3 parts of European plum seed oil and 1.2-6.4 parts of sea sandalwood seed oil;

the plant wax ester is a jojoba ester;

the sterol is at least one of rice chaff sterol and soyasterol;

the squalene is at least one of plant squalene and plant squalane;

the tocopherol is at least one of soybean tocopherol, palm tocotrienol and rice chaff tocotrienol.

The natural vegetable fat compound formed by the simulated human sebum lipid is characterized by comprising the following components in parts by weight: 54-58 parts of vegetable triglyceride, 23-27 parts of vegetable wax ester, 4.3-4.7 parts of sterols, 11-13 parts of squalene and 0.2-0.7 part of tocopherols.

The natural vegetable fat compound formed by the simulated human sebum lipid is characterized by comprising the following components in parts by weight: 57.5 parts of vegetable triglyceride, 26 parts of vegetable wax ester, 4.5 parts of sterols, 12 parts of squalene and 0.5 part of tocopherols.

The preparation method of the natural vegetable fat compound formed by the simulated human sebum lipid is characterized by comprising the following steps:

1) uniformly mixing the meadowfoam seed oil, the macadamia nut oil, the camelina sativa seed oil, the European plum seed oil, the seabuckthorn seed oil and the squalene at a temperature of below 20 ℃ in proportion to obtain a mixed material 1;

2) mixing sterols and tocopherols at a certain proportion, and heating and dissolving at 60-70 deg.C to obtain a mixed material 2;

3) mixing and stirring the mixed materials 1 and 2 at 40-50 ℃, cooling to 35 ℃, adding the vegetable wax ester according to the proportion, stirring uniformly, and cooling to normal temperature to obtain the natural vegetable oil compound.

The natural vegetable oil and fat compound formed by simulating sebum lipid of a human body is characterized in that the vegetable squalene is prepared by the following method:

1) preparing crude mixed oil: sun drying at least two of fructus Canarii albi, semen glycines and semen Tritici Aestivi, pulverizing to 90-100 mesh, squeezing at 40-45 deg.C under 50-60MPa for 65-75min, and filtering to obtain crude mixed oil;

2) and (3) separating and purifying squalene: separating the crude mixed oil by silica gel column chromatography, performing gradient elution by using a mixed solution of n-hexane or triethyl citrate containing 5-8% of ethyl acetate as a mobile phase, controlling the elution flow rate to be 11-13mL/min, collecting and recovering the solvent from each 20mL fraction of the obtained eluent, evaporating the collected 26-28 fraction eluent to remove the solvent, and performing chromatographic detection to obtain the squalene with the purity of 97.96% and the average yield of 86.21%.

The natural vegetable fat and oil compound formed by the human body sebum lipid imitation is applied to the preparation of cosmetics.

The natural vegetable oil-fat compound imitating the sebum lipid of a human body is applied to the preparation of cosmetics, wherein the content of the compound is 1-30%, preferably 8-25%, and more preferably 10-20%.

The meadowfoam seed oil contains long-chain fatty acid with an anti-oxidation effect and a large amount of vitamin E, is a natural antioxidant, can improve the self-resistance of skin, enables the skin to keep soft for a long time, is compounded with other oils, and can also increase the stability of other oils.

The macadamia nut oil contains nutrients which are necessary for forming an oil protective layer on the skin, and most importantly, the macadamia nut oil is mild in oiliness and does not irritate the skin; the oil has good ductility, greasy feeling, good permeability and high solubility to various essential oils, and is a good base oil.

The camelina sativa seed oil contains rich essential fatty acid-omega-3, tocopherol, palmitic acid and other components, is mild and non-irritant to skin, and can improve the content of skin fat, enhance the skin barrier function, prevent photoaging, reduce skin inflammation and other problems.

The European plum seed oil is used as a skin conditioner in cosmetics, can deeply nourish the skin, arouse the vitality of the skin, fade dark yellow, effectively improve the problem of the skin and enable the skin to be fine and smooth.

The rosewood seed oil contains abundant unsaturated fatty acid and vitamin C, is beneficial to keeping the integrity of cell walls, has a recombination effect and an anti-aging effect on skin, and has the functions of bringing good affinity to long-chain fatty acid, increasing the moisture content of the skin, improving sebaceous gland tissues, anti-inflammatory property and improving the blood circulation of the skin.

The jojoba esters are not easy to oxidize, resistant to high temperature and high pressure, small in viscosity change, good in touch and ductility, easy to absorb by skin, and are excellent oil, moisturizer and humectant in cosmetics, the risk coefficient is 1, and the jojoba esters are safe and can be used safely.

The rice chaff sterol is a specific component in rice, has high permeability on skin due to water locking and moisturizing, can maintain the softness and the moisture of cells, and effectively keeps the moisture on the surface of the skin; the rice chaff sterol can also promote skin metabolism, inhibit skin speech, and prevent erythema solare and skin aging.

The soyasterol has antioxidant effect, can maintain the softness and moisture of cells, has mild permeability to skin, can keep the moisture on the surface of skin, promotes skin metabolism, and has the effects of inhibiting epidermal inflammation, delaying skin aging, eliminating color spots and erythema solaris, and the like.

The plant squalene has the following effects: liver protection, promoting hepatocyte regeneration and protecting hepatocyte, thereby improving liver function; efficacy against fatigue of said squalane: strengthen and repair the epidermis, effectively form the natural protective film, help the balance between skin and sebum.

The plant squalene can adopt at least two of olive, soybean and wheat as raw materials, combines the cold pressing method and the silica gel column chromatography method, has the advantages of simple and convenient operation, environmental protection and the like, and can obtain a squalene product with high purity and high yield; in the method, the crude oil is prepared by adopting a cold pressing technology, so that the squalene component contained in the crude oil is not easy to be oxidized; during separation and purification, the types, concentrations and elution speeds of the stationary phase and the mobile phase have obvious influence on the purity and yield of the final product squalene. The wheat germ oil is cereal germ oil prepared from wheat malt, concentrates the nutrition essence of wheat, is rich in squalene, vitamin E, linoleic acid, linolenic acid, octa-carbon alcohol and various physiological active components, and is known as functional oil with nutrition and health care effects. The olive and the soybean also contain squalene and can be used as raw materials for extracting plant squalene. The plant squalane prepared by the hydrogenation of the plant squalene can be directly used in the raw material formula of the invention.

The plant squalene disclosed by the invention adopts at least two of olive, soybean and wheat as raw materials, contains trace other effective components of mixed raw materials besides the extracted high-purity squalene, and has a more remarkable effect due to the synergistic ratio of the effective components in the extract.

The plant squalane is extracted from plants, has good oxidation resistance and thermal stability, is lipid closest to sebum of a human body, has strong affinity, can be integrated with the sebum membrane of the human body, forms a natural barrier on the skin surface, can inhibit the peroxidation of skin lipid,

effectively permeate into skin, promote the proliferation of basal cells of the skin and delay skin aging; squalane can also open skin pores, promote blood circulation, promote cell metabolism, and repair damaged skin.

The tocopherol is a hydrolysate of vitamin E, can increase the antioxidation of cells, has a certain anti-aging effect, and can improve lipid metabolism.

The meadowfoam seed oil, the macadamia nut oil, the camelina sativa seed oil, the European plum seed oil, the seabuckthorn seed oil, the jojoba esters, the rice chaff sterols, the soybean sterols, the plant squalene, the plant squalane, the soybean tocopherols, the palm tocotrienols and the rice chaff tocotrienols can be purchased from the market, and can also be prepared by adopting the method or a conventional extraction method.

As a natural oil compound, the natural oil compound has different components and contents, has obvious difference in efficacy, different raw materials, different values and different utilization rates, and the types and contents of the obtained effective components are different. The limitations of the components and the contents in the natural vegetable oil compound, the selection of the process steps and the process parameters in the preparation process are the optimal values screened by the applicant through a large number of tests, optimization, comparison and analysis, and the effective components in the components are mutually compatible and coordinated, so that the optimal human body sebum lipid imitation effect is achieved.

The natural vegetable oil-fat compound imitating the formation of human sebum lipid and the preparation method and the application thereof construct a stable, mild and non-irritant natural vegetable oil-fat composition, simulate the formation of human sebum and the proportion of fatty acid, thereby obtaining a cosmetic beautifying oil-fat component with high biocompatibility and high bioavailability, being beneficial to enhancing the oxidation protection power of skin, softening and firming skin, maintaining the stable state of skin and achieving the effect of fading fine wrinkles.

Drawings

FIGS. 1-2 are graphs of mean values of tissue viability for example 3 of the present invention, respectively;

FIG. 3 is a diagram showing the relative carbon atom ratios of CERs in example 3 of the present invention;

FIG. 4 is a diagram showing the relative occupation ratio of each of the CER subclasses in example 3 of the present invention;

FIG. 5 is a chart of the average carbon chain length of CERs of example 3 of the present invention;

FIG. 6 is a CER peak area/FFA peak area chart in example 3 of the present invention;

FIG. 7 is a graph showing the trend of the change in tissue viability in example 3 of the present invention;

FIG. 8 is a diagram showing the change of tissue morphology in example 3 of the present invention;

FIG. 9 is a graph showing the trend of the change in the thickness of the stratum corneum of example 3 of the present invention;

FIG. 10 is a graph showing the change in the content of CD44 in example 3 of the present invention;

FIG. 11 is a graph showing the trend of CD44 content in example 3 of the present invention;

FIG. 12 is a graph showing the malondialdehyde content in example 3 of the present invention.

Detailed Description

The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments of the present invention, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. The following described embodiments are exemplary and are intended to be illustrative of the invention and are not to be construed as limiting the invention.

Example 1: 13 parts of meadowfoam seed oil, 40 parts of macadamia nut oil, 2.3 parts of camelina sativa seed oil, 2 parts of European plum seed oil, 2.5 parts of rosewood seed oil, 20 parts of jojoba esters, 4 parts of rice chaff sterol, 2 parts of plant squalene and 10 parts of plant squalane.

Example 2: 18 parts of meadowfoam seed oil, 32 parts of macadamia nut oil, 2 parts of camelina sativa seed oil, 1.8 parts of European plum seed oil, 6 parts of seabuckthorn seed oil, 23 parts of jojoba esters, 6 parts of soyasterol, 1 part of plant squalene, 9.5 parts of plant squalane, 0.2 part of soybean tocopherol, 0.2 part of palm tocotrienol and 0.1 part of rice chaff tocotrienol.

Example 3: 23 parts of meadowfoam seed oil, 28.17 parts of macadamia nut oil, 0.29 part of camelina sativa seed oil, 0.29 part of European plum seed oil, 5.75 parts of seabuckthorn seed oil, 26 parts of jojoba esters, 4.5 parts of rice chaff sterol, 11.5 parts of plant squalane and 0.5 part of palm tocotrienol.

Example 4: 14.66 parts of meadowfoam seed oil, 37.65 parts of macadamia nut oil, 1.73 parts of camelina sativa seed oil, 1.73 parts of European plum seed oil, 1.73 parts of seabuckthorn seed oil, 26 parts of jojoba esters, 4.5 parts of soyasterol, 3.5 parts of plant squalene, 8 parts of plant squalane and 0.5 part of soybean tocopherol.

Example 5: 24 parts of meadowfoam seed oil, 25 parts of macadamia nut oil, 0.18 part of camelina sativa seed oil, 0.2 part of European plum seed oil, 1.2 parts of seabuckthorn seed oil, 30 parts of jojoba esters, 4 parts of rice chaff sterol, 0.5 part of soyasterol, 5 parts of plant squalene, 6.8 parts of plant squalane, 0.1 part of soybean tocopherol, 0.1 part of palm tocotrienol and 0.1 part of rice chaff tocotrienol.

The preparation method of the natural vegetable fat and oil compound comprises the following steps:

The plant squalene is prepared by the following method:

2) and (3) separating and purifying squalene: separating the crude mixed oil by silica gel column chromatography, performing gradient elution by using n-hexane or triethyl citrate containing 5-8% of ethyl acetate as a mobile phase, controlling the elution flow rate to be 11-13mL/min, collecting and recovering the solvent from each 20mL fraction of the obtained eluent, evaporating the collected 26-28 fraction eluent to remove the solvent, and performing chromatographic detection to obtain the squalene with the purity of 97.96% and the average yield of 86.21%. The plant squalane prepared by the hydrogenation of the plant squalene can be directly used in the raw material formula of the invention.

The application of the natural vegetable fat compound formed by the simulated human sebum lipid in the preparation of cosmetics; wherein the content of the complex is 1-30%, preferably 8-25%, more preferably 10-20%.

The design principle of the present invention is illustrated below from the analysis of lipids in the epidermis of a healthy human body. The skin surface lipid secreted by a healthy human body is composed of:

the triglyceride and free fatty acid distribution of the human body was analyzed as:

the invention simulates the constitution of skin surface lipid respectively through the components of various plant source oil and fat:

in order to simulate the fatty acid composition of skin surface lipid, the fatty acid communication of different vegetable oils and fats needs to be analyzed to prepare a proper proportion which is as close to the composition of natural lipid of human body as possible. Taking embodiment 3 of the present invention as an example, the following is specific:

according to the embodiment 3 of the invention, by matching different combinations of the vegetable oils, the component can simulate the distribution of fatty acid in sebum of a human body as much as possible, so that the cosmetic beautifying oil component with high biocompatibility and high bioavailability is obtained. The complexes of the present invention described in examples 1-2 and examples 4-5, also close to the composition of natural lipids in humans.

The beneficial effects of the present invention are further illustrated below in conjunction with other experimental data.

Test one: human test data test.

1. Test method

1.1 measurement of skin hydration

Corneometer MPA 10(Courage and Khazaka, Cologne, Germany) records the capacitance at the skin surface. Capacitance is expressed in digital form in arbitrary units (a.u.). A probe tip (7x7 mm) consisting of a condenser was applied to the skin surface at a constant pressure (3.5 newtons). The measurement principle is based on the significantly different dielectric constants of water (about 81) and most other materials (less than 7). Five measurements were made in each test area and the average was used to define the hydration state of the stratum corneum. The corneometer used in the invention. S/N09372310; probe S/N09341841.

1.2 measurement of biomechanical Properties (elasticity, hardness)

The biomechanical properties of the skin were assessed using a Cutomer MPA580(Courage + Khazaka Electronic GmbH, Cologne; S/N32041888 tube: S/N04317556).

The measurement is based on the vacuum suction principle. By applying a constant negative pressure over a certain period of time, the skin is sucked into a hollow tube with a hole of 2 mm in diameter. The skin is then allowed to retract under normal air pressure. The depth of penetration of the tube by the skin is optically recorded without friction and mechanical influence. Some normalized parameters may be calculated from the resulting penetration depth curve. Most parameters are a function of skin thickness and therefore cannot be simply compared between subject and region.

To improve accuracy and capture information about the skin properties under repeated external pressure, the cycle is repeated several times, and the parameters selected for evaluation are based on the area rather than a single measurement point.

1.2.1 skin firmness

The skin firmness is evaluated by a parameter F4, i.e. the area under the approximate envelope function of the maximum extension. A decrease in F4 corresponds to an increase in skin firmness.

1.2.2 skin elasticity

Skin elasticity was evaluated by the ratio F3/F4. The larger F3 compared to F4, the greater the restoring force and the smaller the residual deformation remaining. The closer the result value is to 1, the greater the elasticity of the skin.

The parameters were calculated by WinCT (Courage & Khazaka GmbH, Colon-Germany).

1.3 measurement of skin roughness

PRIMOS (phase shift rapid in vivo skin measurement) is a non-contact measurement device that can perform real-time three-dimensional in vivo measurements of the micro-topography of human skin based on active image triangulation techniques. The measuring head comprises a digital micromirror device as a projection unit and a CCD camera as a recording unit, mounted on an adjustable frame. For active image triangulation, an intensity encoded point M is projected onto the surface under test. Its image on the surface is recorded by a CCD camera from a particular angle. The point M is a function of parameters such as intensity, triangulation angle between the projection system and the camera, and other camera and projection plane internal and external coordinates. Height information of the structured surface is encoded in a distorted intensity pattern, which is recorded. Resolution and accuracy depend on the optical and topographical features of the surface being measured and the noise characteristics of the measurement system. In order to accurately measure the human skin in vivo, different effective wavelength and amplification factor parameters should be used according to the measured part (inner forearm, forehead, eye region).

In order to take into account differences in human skin and to avoid unnecessary distortions due to motion, measurements were made using a fast phase shift technique (phase width: 16 and 64 pixels). At least 3 recordings are made for each measurement, and the sharpest image without motion distortion or artifacts is selected for further processing.

At the end of the invention, the distortions due to body hair are digitally removed and the macrostructure (calculated by polynomial approximation), i.e. the curvature of the whole test area, is subtracted in order to correctly analyze the microstructure, i.e. the surface roughness.

The skin roughness is then evaluated by the parameter RZ (average roughness). To mitigate potential directional effects, the evaluation employed the arithmetic mean of RZ for 32 radial cuts.

The average roughness is defined as:

where n is the number of equal segments into which the scan length l is divided and RZi is the maximum peak-to-valley depth in each segment. According to the German standard Din 4768/1, RZ is calculated from 5 equal-length segments. The system used in the present invention. PRIMOS compact high resolution S/N108 + 00042, software version 5.7.

1.4 measurement of wrinkle depth/area/volume

As described above, the crow's feet area is recorded as a three-dimensional topographic map using the PRIMOS system. For accurate detection of deeper structures, different fast phase shift settings (phase widths: 16, 64 and 128 pixels) are used. For each measurement, at least 3 recordings are made and the sharpest image without motion distortion or artifacts is selected for further processing. In subsequent visits, the originally captured data was superimposed onto the volunteer's skin to aid in the repositioning process of the test area.

At the end of the invention, the distortions due to body hair were digitally removed and the macrostructure (calculated by polynomial approximation), i.e. the curvature of the whole tested area, was subtracted in order to make a suitable analysis of the microstructure, i.e. wrinkles and surface roughness. For evaluation by the "wrinkle analysis module" of the PRIMOS software, the three-dimensional data from subsequent visits were additionally elastically matched (aligned by movement, rotation, tilting) and the largest common area in the data set for each subject was determined. Thus, the area covered by the analysis varies between different subjects, and therefore absolute wrinkle volume and area measurements should not be compared between subjects.

The resulting data set was then analyzed using classical parameters cut into from the surface and the "wrinkle analysis module" in the PRIMOS software. The following parameters were calculated to characterize the wrinkles.

Wrinkle depth (Rmax, μm): RMax is defined as the maximum vertical distance from the highest peak to the lowest valley.

Wrinkle depth (Rmax, μm): RMax refers to the maximum vertical distance from the highest peak to the lowest valley of five equal lengths. To mitigate the localization effect, the arithmetic mean of Rmax of 50 parallel incisions perpendicular to the main wrinkle across the entire measurement area (0.5 mm border) was used for the evaluation.

Average wrinkle depth (μm): an average wrinkle depth calculated by the wrinkle analysis module.

The average wrinkle depth calculated by the wrinkle analysis module (manual rejection of detection artifacts).

Total area of wrinkles (mm)²): a total measured area covered by wrinkles detected by the wrinkle analysis module.

The total measured area covered by wrinkles detected by the wrinkle analysis module (excluding artifacts detected manually).

Total volume of wrinkles (mm)³) Total volume of wrinkles detected by the wrinkle analysis module (detection artifact excluded manually).

Total volume of wrinkles detected by the wrinkle analysis module (excluding artifacts detected manually). Can be calculated from the average depth and the area covered.

The system used in the present invention. PRIMOS compact high resolution S/N108 + 00041, software version 5.7.

2. The results of the tests are shown in Table 1.

TABLE 110% Natural vegetable fat Complex (example 3) data test Table for human test data of caprylic/capric triglyceride solution

3. Conclusion of the test

Human body test data prove that the composition can obviously improve the moisture content of the skin, improve the smoothness of the skin, improve the elasticity of the skin and reduce wrinkles after being tried out for 28 days.

Skin moisture content: the test samples were found to statistically significantly increase skin hydration. Over 28 days of use, an average increase of 23.1% was observed, with effects on 100% of volunteers.

Skin firmness and elasticity: the test sample can obviously improve the elasticity of the skin, so that the optimal state of tightness and elasticity is achieved. Over 28 days of use, the effect of the test product was detected in volunteers with an average increase of 18.1% (firmness)/9.8% (elasticity), 100% (firmness)/80% (elasticity).

Skin smoothness test products were found to statistically significantly increase the smoothness of the skin. Over 28 days of use, an average increase of 14.5% was observed, with an effect on 85% of volunteers.

Depth, area and volume of wrinkles in conventional use, the test product was found to reduce wrinkle depth, area and volume statistically significantly. After 28 days of treatment, a reduction in mean depth of 13% (classical cutting analysis)/11% (wrinkle analysis module) was observed, with effective fruits in 95% of volunteers.

The beneficial effects of the present invention can also be achieved by performing the above human test data tests using examples 1-2 and examples 4-5.

And (2) test II: in vitro test data assay.

1. Introduction of indexes

1.1 tough barrier

1.1.1 Barrier index

The barrier index ET50 is an index that reflects the degree of skin tolerance to irritants. And after the administration and incubation of the sample are finished, TritonX-100 is acted on the surface of the model for 0h, 1h and 3h, the tissue viability is detected after the action is finished, and the ET50 value is calculated according to the tissue viability at 3 points. The higher the ET50 value, the stronger the barrier resistance.

1.1.2 Barrier lipids

Ceramides and fatty acids are the major constituents of barrier lipids, and ceramides, fatty acid subclass constituents (saturated fatty acids, unsaturated fatty acids, hydroxylated fatty acids) and the ratio of ceramides and fatty acids affect skin barrier structure and penetration function.

1.2 epidermal anti-aging (anti-aging barrier)

1.2.1 tissue viability

The vitality of skin cells is closely related to the exertion of normal functions of skin, and functional damage is firstly related to the decline of the vitality of the skin cells, so as to cause a series of problems of inflammation, aging, even skin diseases and the like. Increasing cell viability helps the skin to resist adverse environmental effects.

1.2.2 CD44

CD44 is a receptor for hyaluronic acid, and is capable of binding hyaluronic acid to exert a number of biological effects, such as moisturizing, skin barrier remodeling; with age, the level of CD44 in the skin gradually decreases. Therefore, the content of CD44 in the skin is increased, the skin moisturizing capability can be improved, the skin is full and plump, the utilization rate of exogenous hyaluronic acid can be effectively increased, and the capacity of barrier reconstruction of aged skin is increased.

1.2.3 tissue morphology

Tissue morphology is the microscopic physiological structural analysis of the tissue after H & E staining. Through the histological morphology of the model, the change condition of the skin structure under different treatment conditions can be intuitively known, and the stratum corneum is very obviously seen by the model with weakened barrier, so that the barrier lifting function of the sample to be tested can be evaluated through the change of the thickness of the stratum corneum.

1.3 Oxidation resistance (lipid peroxidation protection)

1.3.1 cell viability

Keratinocytes are the main cell type of the epidermis. Oxidative stress caused by the environment and the inside can cause the peroxidation of cell lipids, thereby damaging the viability and normal physiological functions of cells. The cells can evaluate the protective ability of the sample to be tested on lipid oxidation through the external oxidative stress.

2. Purpose of testing

The efficacy of this composition (the test sample in this experiment is example 3) in the robust barrier, epidermal anti-aging, protection against lipid peroxidation on in vitro recombinant 3D skin tissue models and keratinocyte models was determined.

3. Experimental Material

3.1 test System

The skin model used in the test is a 3D epidermal skin model

(hereinafter referred to as skin model), the batch number is: ES2000806, manufactured and supplied by Guangdong Boxi Biotech Co.

3.2 Primary reagents

TA culture medium (special culture medium for skin model), phosphate buffer solution (solibao), mtt (sigma), isopropyl alcohol (national drug), 4% paraformaldehyde (BioSharp), xylene (national drug), absolute ethyl alcohol (national drug), hematoxylin (bi yun day), eosin (bi yun day), hydrochloric acid (national drug), 50 × sodium citrate (siennatt bio), Anti-CD44 antibody (Abcam),

ABC-Peroxidase Kits (VECTASTAIN), Anti-Mouse-488 (goat Anti Mouse) (Abcam), Anti-quencher (Biyun day), Hochest33342 (Biyun day).

3.3 Main Equipment

CO2 incubator (Thermo), clean bench (sujing altai), microplate reader (BioTek), micro-oscillator (its linbel), upright microscope (Olympus).

4. Scheme design, see table 2.

TABLE 2 in vitro test protocol design Table (sample: EXAMPLE 3)

5. Results of the experiment

5.1 Barrier improvement efficacy test results

Experiment grouping

Remarking: firstly, performing ET50 detection on BC group TA 7; the PC group and the test sample group are subjected to administration treatment by TA7, and the TA8 is subjected to ET50 detection. ② PC group as positive control, WY14643(PPAR agonist) was administered.

5.1.1 Barrier index test results, see FIGS. 1 and 2.

In fig. 1: when statistically analyzed by the t-test method, the significance was expressed as P-value <0.05 and P-value < 0.01, compared to the BC group. Fig. 1 shows that the tissue viability of the PC (WY14643) group was significantly increased at 0h, 1h and 3h compared to the BC group, indicating that this positive control assay was effective; the tissue viability of the test sample groups was significantly increased at 0h, 1h and 3 h.

In fig. 2: according to the result of the ET50 value, the ET50 value of the sample group is higher than the ET50 value of the BC group; therefore, the sample has good barrier lifting efficacy.

5.1.2 Barrier lipid assay results, see FIGS. 3-6.

Fig. 3 shows that the relative proportions of C63, C65, C67 and C69 in the ceramides of the sample group were significantly increased by 222.28%, 244.26%, 378.56% and 264.71%, respectively, compared to the BC group. Fig. 4 shows that the relative percentage of ceramide subclass EOP was significantly increased, 272.52% higher in the sample group compared to the BC group. Figure 5 shows that the mean carbon chain length of the ceramides was significantly increased by 7.35% in the sample group compared to the BC group. Figure 6 shows that the ceramide to fatty acid ratio was significantly increased by 37.56% in the sample group compared to the BC group.

5.2 detection results of epidermal anti-aging efficacy

Experiment grouping

Remarking: PC group was a positive control and was administered WY14643(PPAR agonist).

5.2.1 tissue viability assay results, see FIG. 7.

In fig. 7: when the statistical analysis is carried out by using the t-test method, compared with the BC group, the significance is represented by #, and P-value <0.05 is represented by #, and P-value; < 0.01 as # #; significance was expressed as P-value <0.05 and P-value < 0.01 compared to the NC group. Fig. 7 shows that the tissue viability of the NC group was significantly reduced compared to the BC group, indicating that the stimulation conditions were effective in this experiment. Compared with the NC group, the tissue viability of the PC (WY14643) group was significantly increased, indicating that this experiment was effective. Compared with the NC group, the tissue viability of the sample group is remarkably increased by 6.14%.

5.2.2 tissue morphology results, see FIGS. 8-9.

In FIGS. 8-9: when the statistical analysis is carried out by using the t-test method, compared with the BC group, the significance is represented by #, the P-value <0.05 is represented by #, and the P-value < 0.01 is represented by # #; significance was expressed as P-value <0.05 and P-value < 0.01 compared to the NC group. FIGS. 8-9 show that the average stratum corneum thickness in the NC group was significantly increased compared to the BC group, indicating that the stimulation conditions were effective in this experiment; compared with the NC group, the average thickness of the stratum corneum of the PC (WY14643) group is obviously reduced, which shows that the experiment is effective; the mean stratum corneum thickness of the sample group was significantly reduced by 23.92% compared to the NC group.

5.2.3CD44 immunohistochemical assay results, see FIGS. 10-11.

In FIGS. 10-11: when the statistical analysis is carried out by using the t-test method, compared with the BC group, the significance is represented by #, the P-value <0.05 is represented by #, and the P-value < 0.01 is represented by # #; significance was expressed as P-value <0.05 and P-value < 0.01 compared to the NC group. Fig. 10-11 show that CD44 content in NC group was significantly reduced compared to BC group, indicating that the stimulation conditions were effective in this experiment; compared with the NC group, the content of CD44 in the PC (WY14643) group is obviously increased, which shows that the experiment is effective; compared with the NC group, the CD44 content of the sample group is increased remarkably by 25.85%.

5.3 efficacy test results for prevention of lipid peroxidation, see experimental groups and FIG. 12.

Experiment grouping

In fig. 12: mean SD of five independent experiments, showing statistically significant differences with P < 0.05. Fig. 12 shows that the malondialdehyde content of the positive group was significantly increased compared to the blank group, indicating that the stimulation conditions of this experiment were effective; compared with the blank group, the malondialdehyde content of the control group I is obviously reduced, which indicates that the sample effectively prevents cell lipid peroxidation under the condition of no stimulation; the malondialdehyde content of control group II was significantly reduced (28.5% calculated as malondialdehyde) compared to the positive group, indicating that the sample was effective in preventing lipid peroxidation of cells under oxidative stress.

6. Summary of results

The test result shows that: the natural plant complex (example 3) of the present invention has a barrier enhancing effect by promoting differentiation of epidermal cells and formation of barrier function; meanwhile, the composition has the capability of resisting epidermal aging caused by tissue damage by improving the tissue vitality and increasing the content of CD44 in the skin. The beneficial effects of the present invention can also be achieved by performing the in vitro test data tests described above with examples 1-2 and examples 4-5.

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be able to cover the technical scope of the present invention and the equivalent alternatives or modifications according to the technical solution and the inventive concept of the present invention within the technical scope of the present invention.

Claims

1. A natural vegetable fat compound imitating the constitution of human body sebum lipid is characterized by comprising the following components in parts by weight: 50-60 parts of vegetable triglyceride, 20-30 parts of vegetable wax ester, 4-6 parts of sterols, 11-13 parts of squalene and 0-0.8 part of tocopherols;

the plant wax ester is a jojoba ester;

the sterol is at least one of rice chaff sterol and soyasterol;

the squalene is at least one of plant squalene and plant squalane;

2. The natural vegetable fat and oil composition imitating sebum lipid of human body as claimed in claim 1, wherein the composition comprises the following components in parts by weight: 54-58 parts of vegetable triglyceride, 23-27 parts of vegetable wax ester, 4.3-4.7 parts of sterols, 11-13 parts of squalene and 0.2-0.7 part of tocopherols.

3. The natural vegetable fat and oil composition imitating sebum lipid of human body as claimed in claim 1, wherein the composition comprises the following components in parts by weight: 57.5 parts of vegetable triglyceride, 26 parts of vegetable wax ester, 4.5 parts of sterols, 12 parts of squalene and 0.5 part of tocopherols.

4. The method for preparing a natural vegetable lipid complex imitating the constitution of sebum lipids of human body as claimed in claim 1, comprising the steps of:

5. The natural vegetable oil and fat composition imitating the constitution of sebum lipid of human body as claimed in claim 1, wherein the vegetable squalene is prepared by the following method:

2) and (3) separating and purifying squalene: separating the crude mixed oil by silica gel column chromatography, performing gradient elution by using a mixed solution of n-hexane or triethyl citrate containing 5-8% of ethyl acetate as a mobile phase, controlling the elution flow rate to be 11-13mL/min, collecting and recovering the solvent from each 20mL fraction of the obtained eluent, evaporating the collected eluent of 26-28 fractions to remove the solvent, and performing chromatographic detection to obtain the high-purity squalene.

6. Use of a natural vegetable lipid complex mimicking the sebum lipid composition of human body as claimed in claim 1 in the preparation of cosmetics.

7. Use of a natural vegetable lipid complex mimicking the sebum lipid content of a human body as claimed in claim 6, wherein the complex is present in an amount of 1-30%, preferably 8-25%, more preferably 10-20% for the preparation of a cosmetic product.