Anti-aging repairing composition containing camellia nitidissima extract and application of anti-aging repairing composition

Abstract

The invention relates to an anti-aging repair composition containing a golden camellia extract and application. The liposome composition sequentially comprises a core layer, a shell layer and a chitosan coating from inside to outside, wherein the core layer comprises a camellia nitidissima extract and DNA sodium; the shell layer comprises lecithin, cholesterol, ceramide and an enzymolysis tea extract. Active substances are wrapped by a coating liposome wrapping technology, the active substances are helped to permeate the skin, the skin aging condition is comprehensively improved from multiple dimensions of promoting collagen synthesis, enhancing skin elasticity, resisting oxidation, repairing damaged skin barriers and the like, and the remarkable anti-aging and repairing effects are achieved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of liposomes, and particularly relates to an anti-aging and repair composition containing camellia nitidissima chun extract as well as a preparation method and application thereof. BACKGROUND

[0002] In today's society, with the improvement of people's living standards and the enhancement of health consciousness, anti-aging and repair has become one of the core demands in the field of skin care. Facial skin aging is a complex and progressive process, which is affected by multiple factors such as natural aging, photoaging, unhealthy lifestyle, environmental pollution and mental stress, and is manifested as a series of problems such as increased wrinkles, skin laxity, pigmentation, and decreased barrier function, which seriously affects people's appearance and mental health.

[0003] At present, there are various anti-aging and repair products on the market, among which emulsions are the basic category of daily skin care. However, the existing anti-aging and repair emulsion products still have many defects. On the one hand, the formulations of some products lack scientificity and systematicness, and excessively rely on single ingredients or blindly pile up popular ingredients, which is difficult to comprehensively and effectively solve the problem of skin aging, and even may cause secondary damage to the skin due to incompatibility or irritation between ingredients. Therefore, how to reduce the irritation of the product while ensuring the anti-aging and repair effect and improve the use comfort has become a key problem to be solved for current anti-aging and repair products. On the other hand, the skin barrier often prevents skin care ingredients from penetrating deeply and exerting their efficacy. Therefore, new liposome penetration technology can be developed to solve this problem. Liposomes are similar in structure to cell membranes, can smoothly pass through the intercellular space of keratinocytes, promote the transdermal absorption of active ingredients, and thus improve the bioavailability. In addition, liposomes have a wrapping effect, which can protect active substances, improve the stability and solubility of active substances. However, the liposomes on the market are mostly wrapped with single ingredients, and have poor stability. SUMMARY

[0004] The present application aims to overcome the deficiencies of the prior art and provide an anti-aging and repair composition containing camellia nitidissima chun extract as well as a preparation method and application thereof. The core layer of the liposome composition includes camellia nitidissima chun extract and DNA sodium, and the shell layer includes ceramide and enzymatic tea extract. The use of coating liposome wrapping technology can promote the transdermal absorption of these active ingredients at the same time, thereby exerting better efficacy.

[0005] To achieve the above-mentioned purpose, the technical scheme adopted by the present application is as follows:

[0006] In a first aspect, the present application provides an anti-aging protective composition containing camellia japonica extract, which comprises, from inside to outside, a core layer, a shell layer and a chitosan coating layer; wherein the core layer comprises camellia japonica extract and DNA sodium; the shell layer comprises lecithin, cholesterol, ceramide and enzymatic tea extract; the chitosan coating layer comprises chitosan; and the weight ratio of the camellia japonica extract, DNA sodium, lecithin, cholesterol, ceramide, enzymatic tea extract and chitosan is (0.1-10):(0.001-2):(0.1-10):(0.1-5):(0.001-2):(0.001-2):(0.01-3).

[0007] The functions of the raw materials of the liposome shell layer of the present application are as follows:

[0008] The lecithin, as the main material of the liposome, has dual characteristics of hydrophilicity and lipophilicity, is an important component of cell membrane, can supplement skin lipids, repair damaged stratum corneum and strengthen skin barrier; the cholesterol is an important component of the skin natural barrier, can combine with the lipids in the liposome, strengthen the stability and elasticity of the liposome, and enhance the skin barrier function; the chitosan molecule modifies the surface of the liposome, makes the surface of the liposome positively charged, can improve the stability of the liposome and the adsorption and penetration of the skin, and has strong water absorption, which can repair the skin barrier; the ceramide is the main component of the intercellular lipids of the skin stratum corneum, which can help skin to metabolize aged cells, stimulate the generation of new cells, smooth wrinkles, repair the stratum corneum and play an important role in maintaining the skin barrier function; the enzymatic tea extract releases more tea polyphenols by accurately decomposing the cell wall of tea leaves through enzymatic technology, so that the active ingredients in the tea extract are more easily absorbed by the skin, and the antioxidant and anti-inflammatory effects are exerted, and the skin permeability and stability of the enzymatic tea extract are further improved by being wrapped in the shell layer of the liposome.

[0009] The functions of the raw materials of the liposome core layer of the present application are as follows:

[0010] The flavonoids and tea polyphenols in the camellia japonica extract are the main antioxidants, have the activity of scavenging free radicals, can reduce the damage of oxidative stress to the skin, delay skin aging, enhance skin elasticity and reduce the generation of wrinkles; the DNA sodium (polydeoxyribonucleotide, PDRN) has the core advantages of activating the skin self-repairing ability from the cell level and providing a mild anti-aging solution, and the skin permeability and anti-aging effect of the DNA sodium are improved by being wrapped in the core layer of the liposome.

[0011] The surface of the chitosan-coated liposome is positively charged, and the positively charged chitosan-coated liposome can interact with the negative charge on the surface of the skin, change the secondary structure of the keratin in the stratum corneum, improve the fluidity of the cell membrane, increase the permeability of the skin, and make the effective components in the skin care product more easily absorbed by the skin, thereby improving the efficacy of the product, secondly, the chitosan-coated liposome can play an antibacterial and anti-inflammatory role, reduce the inflammatory symptoms, and promote the healing of the skin, and the antibacterial mechanism is mainly to interact with the negatively charged components on the surface of the bacteria, destroy the cell wall and cell membrane structure of the bacteria, and thus cause the death of the bacteria. In addition, the liposome core layer of the present application wraps the camellia nitidissima extract and DNA sodium, and the shell layer wraps the ceramide and the enzyme-decomposed tea leaf extract, which can simultaneously promote the stability and transdermal absorption of these active ingredients, thereby playing a better effect.

[0012] Preferably, the weight ratio of the camellia nitidissima extract and the DNA sodium is (1-3):(0.01-0.5).

[0013] Preferably, the weight ratio of the lecithin, cholesterol, ceramide, enzyme-decomposed tea leaf extract and chitosan is (1-8):(0.5-3):(0.01-1):(0.01-0.5):(0.1-1).

[0014] Preferably, the weight ratio of the camellia nitidissima extract and the DNA sodium is (1.5-2):(0.05-0.1).

[0015] Preferably, the weight ratio of the lecithin, cholesterol, ceramide, enzyme-decomposed tea leaf extract and chitosan is (4-5):(0.8-1):(0.05-0.1):(0.05-0.1):(0.4-0.5).

[0016] Preferably, the preparation method of the enzyme-decomposed tea leaf extract comprises the following steps:

[0017] (1) raw material pretreatment: fresh, clean and non-rotting withered tea leaves are selected as raw materials, microwave treatment is performed for dehydration and fixation, and after cooling, the dried tea leaves are crushed by a crushing device, sieved, and tea leaf powder is obtained; wherein the power of the microwave treatment is 300-500W, and the treatment time is 40-60s; the particle size of the sieving is 40-60 meshes, and the microwave treatment is used to destroy the enzyme activity in the tea leaves;

[0018] (2) Enzymatic hydrolysis: adding sterilized deionized water to the tea powder, mixing and stirring evenly, adding compound enzyme, and performing enzymatic hydrolysis at a temperature of 35-45°C for 2-5 hours to allow the enzyme to fully decompose the tea cell wall and release the effective ingredients in the tea, and then performing enzyme inactivation treatment on the enzymatically hydrolyzed mixture to obtain an enzymatic hydrolyzate; wherein, the amount of sterilized deionized water added is 6-7 times the weight of the tea powder, and the compound enzyme includes cellulase and pectinase in a weight ratio of 1:1, and the amount of the compound enzyme added is 0.6%-1% of the weight of the tea powder; wherein, the enzyme inactivation treatment is: placing the enzymatically hydrolyzed mixture in a boiling water bath for 5 minutes to inactivate the enzyme and stop the enzymatic hydrolysis reaction;

[0019] (3) Three-stage extraction: adding ethanol solution to the enzymatic hydrolysate, performing the first stage extraction to obtain the first stage extract; performing the second stage extraction on the mixed solution after the first stage extraction to obtain the second stage extract; performing the third stage extraction on the mixed solution after the second stage extraction to obtain the extract; wherein, the mass concentration of the ethanol solution is 45-55%, the weight ratio of the enzymatic hydrolysate to the ethanol solution is 1:(3-5), the first stage extraction temperature is 25-35°C, the time is 35-45min, the second stage extraction temperature is 55-65°C, the time is 35-45min, and the third stage extraction temperature is 75-85°C, the time is 15-25min; through the three-stage extraction, different effective components in tea can be fully extracted and the extraction rate can be improved;

[0020] (4) Extraction liquid treatment: The extract after the multi-stage extraction is subjected to centrifugation and filtration treatment to further purify the active ingredients in the extract to obtain a crude tea extract, which is then subjected to adsorption and desorption treatments using a macroporous resin in sequence. The desorbed extract is then subjected to vacuum distillation treatment to obtain a tea mixture;

[0021] (5) Spray drying: spray drying the tea mixture to obtain the enzymatically hydrolyzed tea extract in powder form; wherein the spray drying parameters are: inlet air temperature of 95-105°C, outlet air temperature of 75-85°C, and feed rate of 10-20 mL / min.

[0022] In a second aspect, the present invention provides a method for preparing the anti-aging repair composition containing the Camellia chrysantha extract of the first aspect, comprising the following steps:

[0023] S1, lecithin, cholesterol, ceramide, enzymatic tea extract, 50ml anhydrous ethanol are added into a round-bottom flask, the round-bottom flask is installed on a rotary evaporator, stirring and water bath heating are started until the components are dissolved to form a lipid solution; the anhydrous ethanol in the lipid solution is evaporated until a thin film of lipid is formed on the inner wall of the flask; wherein the stirring speed is 500-600r / min, the water bath temperature is 35-45℃, the vacuum degree for evaporating anhydrous ethanol is 0.08-0.1MPa, the temperature is 35-45℃, and the amount of anhydrous ethanol added is 2-3 times the mass of lecithin, cholesterol, ceramide and enzymatic tea extract;

[0024] S2, the camellia sinensis var. assamica extract and DNA sodium are dissolved in water to obtain a core layer solution, the core layer solution is added into the round-bottom flask in which a thin film has been formed in step S1, stirring and water bath heating are started until the lipid thin film is completely hydrated to form multilamellar liposomes, and the multilamellar liposomes are subjected to ultrasonic treatment to obtain a liposome suspension; wherein the mass concentration of the core layer solution is 30-40%, the stirring speed is 500-600r / min, and the water bath temperature is 35-45℃;

[0025] S3, chitosan coating: chitosan is dissolved in a 0.1%-1% glacial acetic acid solution to prepare a chitosan solution with a mass concentration of 0.2%-1.0%, the liposome suspension is mixed with the prepared chitosan solution at a ratio of 1:0.5, stirring is performed at 25-30℃, and then standing is performed to allow chitosan to bind to the liposomes through electrostatic interaction and weak hydrophobic force, so that the electrostatic binding is stable, and then centrifugation is performed to obtain chitosan-coated liposomes;

[0026] S4, ultrafiltration: the chitosan-coated liposomes are subjected to ultrafiltration to remove unencapsulated components and free lipids, and the anti-aging and repair composition containing the camellia sinensis var. assamica extract is obtained.

[0027] Preferably, in the preparation step S3 of the anti-aging and repair composition containing the camellia sinensis var. assamica extract, the molecular weight of the chitosan is 200-450kDa; after 1.5 hours of magnetic stirring at 200-400rpm and 25-30℃, standing is performed for 30 minutes, and then centrifugation is performed at a speed of 10000rpm for 10min to remove chitosan that is not bound, and the chitosan-coated liposomes are obtained.

[0028] Preferably, in the preparation step S4 of the anti-aging and repair composition containing the camellia sinensis var. assamica extract, the ultrafiltration parameters are as follows: tangential flow filtration technology is used, a flat membrane bag is used, regenerated cellulose is selected as the membrane material, the pore size is 50nm, the transmembrane pressure (TMP) is 0.1-0.4MPa, and the tangential flow rate is 1-3m / s.

[0029] In a third aspect, the present application provides a use of the anti-aging and repair composition containing the camellia sinensis var. assamica extract in the first aspect in the preparation of a cosmetic product.

[0030] In a fourth aspect, the present invention provides an emulsion comprising the following raw materials in weight percentage: 5%-20% of the anti-aging repair composition containing Camellia chrysantha extract in the first aspect, 0.05%-0.5% thickener, 1%-10% moisturizer, 1%-6% emulsifier, 5%-15% oil, 0.5%-3% preservative and 0.01%-0.3% pH regulator, and the balance being deionized water.

[0031] Preferably, the thickener includes at least one of xanthan gum, carbomer, hydroxyethyl acrylate / sodium acryloyldimethyl taurate copolymer, ammonium acryloyldimethyl taurate / VP copolymer, hydroxyethyl cellulose, and cetyl alcohol.

[0032] Preferably, the moisturizing agent includes at least one of allantoin, sodium polyacrylate, panthenol, β-glucan, trehalose, caprylyl glycol, dipropylene glycol, sodium hyaluronate, 1,2-butylene glycol, glycerin, and tremella polysaccharide.

[0033] Preferably, the pH adjuster comprises at least one of arginine, citric acid, NaOH, and disodium EDTA.

[0034] Preferably, the emulsifier includes at least one of PEG-100 glyceryl stearate, glyceryl stearate citrate, cetearyl glucoside, cetyl alcohol, stearyl alcohol, cetearyl alcohol, PEG-20 methyl glucose sesquistearate, methyl glucose sesquistearate, potassium cetyl phosphate, and sodium stearoyl glutamate.

[0035] Preferably, the oil comprises at least one of caprylic / capric triglyceride, dimethicone, jojoba oil, grape seed oil, meadowfoam seed oil, squalane, macadamia nut oil, and camellia oil.

[0036] Preferably, the preservative includes at least one of 1,2-hexanediol, 1,2-pentanediol, ethylhexylglycerin, p-hydroxyacetophenone, phenoxyethanol, caprylhydroxamic acid, and sodium benzoate.

[0037] In a fifth aspect, the present invention provides a method for preparing the emulsion of the fourth aspect, comprising the following steps:

[0038] (1) Mix the thickener and the moisturizer evenly, add deionized water, heat to 70-80°C, homogenize, and keep warm for later use to obtain component A;

[0039] (2) Mix the oil and emulsifier, heat to 70-80°C, stir to dissolve, and keep warm for later use to obtain component B;

[0040] (3) Heat component A to 70-80°C, add component B, and homogenize to obtain an emulsified base.

[0041] (4) The emulsifying base is stirred and cooled to 30-50℃, the anti-aging repair composition containing the camellia nitidissima chung extract is added, the stirring is continued until the material is uniform, the preservative and the pH regulator are added to adjust the pH to 6.0-6.5, the stirring is stopped, the material is discharged, and the emulsion is obtained.

[0042] Compared with the prior art, the anti-aging repair composition has the following beneficial effects:

[0043] The purpose of the present application is to provide an anti-aging repair composition, which comprises a core layer, a shell layer and a chitosan coating layer from inside to outside, wherein the core layer comprises a camellia nitidissima chung extract and DNA sodium, and the shell layer comprises ceramide and an enzymatic tea extract. The active ingredients are wrapped by the coating liposome encapsulation technology to realize transdermal absorption, so that the liposome composition of the present application can comprehensively improve the skin aging condition from multiple dimensions such as promoting collagen synthesis, enhancing skin elasticity, antioxidant, repairing damaged skin barrier, and realizing significant anti-aging repair effect. BRIEF DESCRIPTION OF DRAWINGS

[0044] Figure 1 It is a comparison chart before and after the use of the emulsion of application example 1.

[0045] In order to better illustrate the purpose, technical scheme and advantages of the present application, the present application will be further described below in combination with specific examples.

[0046] The raw materials used in the following examples and comparative examples are as follows:

[0047] Camellia nitidissima chung extract: manufacturer is Ximu Source (Xiamen) Biotechnology Co., Ltd., model is camellia nitidissima chung extract;

[0048] Marigold extract: manufacturer is Xi'an Lvtian Biotechnology Co., Ltd., model is PBJ0301;

[0049] DNA sodium: Shenzhen Weiqi Technology Co., Ltd., model is DNA sodium;

[0050] Ceramide: Shanghai Yuanye Biotechnology Co., Ltd., model S35454-25mg;

[0051] Tea: Wuyishan Wumengjikai Tea Co., Ltd., model is green tea;

[0052] Cellulase: Yinglian Enzyme Preparation Trade (Shanghai) Co., Ltd., model is CL;

[0053] Pectinase: Ningxia Xiasen Industrial Group Co., Ltd., model is FFY-0654.

[0054] Other materials and reagents used in the examples can be obtained from commercial channels unless otherwise specified.

[0055] The preparation method of the enzymatic tea extract includes the following steps:

[0056] The preparation method of the enzymatic tea extract includes the following steps:

[0057] (1) Raw material pretreatment: Fresh, clean, and non-rotting and withering tea leaves are selected as raw materials, and microwave treatment is performed for dehydration and fixation. After cooling, the dried tea leaves are crushed using a crushing device, and sieved to obtain tea powder. The power of the microwave treatment is 400 W, and the treatment time is 50 s. The particle size of the sieving is 40-60 mesh. The microwave treatment is used to destroy the enzyme activity in the tea leaves.

[0058] (2) Enzymatic treatment: Sterilized deionized water is added to the tea powder, and after uniform mixing and stirring, a composite enzyme is added. The enzyme is allowed to fully decompose the cell wall of the tea leaves at a temperature of 40°C for 4 h to release the effective components in the tea leaves. The mixture after enzymatic treatment is subjected to enzyme inactivation treatment to obtain an enzymatic solution. The amount of sterilized deionized water added is 7 times the weight of the tea powder. The composite enzyme includes cellulase and pectinase in a weight ratio of 1:1, and the amount of composite enzyme added is 0.8% of the weight of the tea powder. The enzyme inactivation treatment involves placing the mixture after enzymatic treatment in a boiling water bath for 5 min to inactivate the enzyme and stop the enzymatic reaction.

[0059] (3) Three-stage extraction: Ethanol solution is added to the enzymatic solution for first-stage extraction to obtain a first-stage extract. The mixture after the first-stage extraction is subjected to second-stage extraction to obtain a second-stage extract. The mixture after the second-stage extraction is subjected to third-stage extraction to obtain an extract. The mass concentration of the ethanol solution is 50%, and the weight ratio of the enzymatic solution to the ethanol solution is 1:4. The first-stage extraction temperature is 30°C, and the time is 40 min. The second-stage extraction temperature is 60°C, and the time is 40 min. The third-stage extraction temperature is 80°C, and the time is 20 min.

[0060] (4) Extract treatment: centrifugation and filtration are performed on the extract after multi-stage extraction to further purify the effective components in the extract, to obtain a crude tea extract. The crude tea extract is subjected to adsorption and desorption purification treatment with a macroporous resin in sequence. Specifically, AB-8 type weakly polar macroporous resin is used for purification. First, the crude extract is adjusted to a pH of 3.0-4.0 and filtered, and then loaded onto a pretreated resin column (resin dynamic adsorption capacity: 80-100 mg tea polyphenol / mL) at a flow rate of 2-3 BV / h. When the tea polyphenol concentration of the effluent reaches 1 / 5-1 / 10 of that of the loading solution, the adsorption is stopped. Then, deionized water is used to flush at a flow rate of 3-4 BV / h until the conductivity is ≤50 μS / cm to remove macromolecular impurities. Then, 20% ethanol (2 BV, 2 BV / h), 30% ethanol (3 BV, 1.5 BV / h), and 50% ethanol (4 BV, 1 BV / h) are used in sequence for gradient elution. The tea extract is obtained by combining the elution sections of 30% and 50% ethanol. The resin can be reused after being washed with 70% ethanol and regenerated by acid and alkali.

[0061] (5) Spray drying: the eluate is concentrated (≤50℃) to a solid content of 20%-30% under reduced pressure, and then spray dried. The parameters for spray drying are as follows: inlet air temperature: 160-180℃, outlet air temperature: 70-90℃. Finally, a powdered tea extract is obtained.

[0062] Examples 1-7 and Comparative Examples 1-5

[0063] The mass ratio between the components of the anti-aging protective composition of Examples 1-7 and Comparative Examples 1-5 is shown in Table 1. The total mass of the anti-aging protective composition is 100 parts.

[0064] The preparation method of the anti-aging protective composition of Examples 1-7 and Comparative Examples 1-5 comprises the following steps:

[0065] S1, lecithin, cholesterol, ceramide, enzymatic tea extract, and anhydrous ethanol are added to a round-bottom flask. The round-bottom flask is installed on a rotary evaporator, and stirring and water bath heating are turned on until the components are dissolved, forming a lipid solution. The anhydrous ethanol in the lipid solution is evaporated until a thin film of lipid is formed on the inner wall of the flask. The stirring speed is 550 r / min, the water bath temperature is 40℃, the evaporation pressure of anhydrous ethanol is 0.09 MPa, and the evaporation temperature of anhydrous ethanol is 40℃. The amount of anhydrous ethanol added is 2 times the mass of lecithin, cholesterol, ceramide, and enzymatic tea extract.

[0066] S2, dissolve the Camellia leaf extract and DNA sodium in water to obtain a core layer solution, add the core layer solution to the round-bottom flask in which the thin film has been formed in step S1, start stirring and water bath heating to completely hydrate the lipid film, form multilamellar liposomes, and then perform ultrasonic treatment on the multilamellar liposomes to obtain a liposome suspension; wherein the mass concentration of the core layer solution is 35%, the stirring speed is 550 r / min, the water bath temperature is 40°C, the ultrasonic treatment frequency is 20 KHz, the power is 500 W, and the time is 20 min;

[0067] S3, chitosan coating: select chitosan with a molecular weight of 200-450 kDa: concentration 0.2%-1.0% (dissolved in 0.1%-1% glacial acetic acid), 200-400 rpm magnetic stirring for 12 hours to completely dissolve, filter sterilization (0.22 μm filter membrane), mix the liposome suspension with the prepared chitosan solution at a ratio of 1:0.5, and then magnetically stir at 200-400 rpm for 1.5 hours at 25-30°C, and then stand for 30 minutes to stabilize the electrostatic combination, centrifuge (10,000 rpm, 10 minutes) to remove the unbound chitosan, and obtain the chitosan-coated liposomes;

[0068] S4, ultrafiltration: the chitosan-coated liposomes are subjected to ultrafiltration, specifically: using tangential flow filtration technology, using a flat membrane package, the membrane material is regenerated cellulose, the pore size range is 50 nm, the transmembrane pressure (TMP) is controlled at 0.1-0.4 MPa, the tangential flow rate is 1-3 m / s, and the unencapsulated components and free lipids are removed to obtain the anti-aging and repair composition containing the Camellia leaf extract.

[0069] Table 1: weight ratio of each raw material in the anti-aging and repair composition of each group

[0070]

[0071]

[0072] Comparative Example 6

[0073] Comparative Example 6 differs from Example 1 in that the Camellia leaf extract in Comparative Example 6 is replaced by a marigold extract.

[0074] Comparative Example 7

[0075] Comparative Example 7 differs from Example 1 in that the enzyme-degraded tea leaf extract in Example 1 is replaced by a tea leaf extract in Comparative Example 7. The preparation method of the tea leaf extract comprises the following steps:

[0076] (1) Raw material pretreatment: fresh, clean, non-rotting and withering tea leaves are selected as raw materials, microwave treatment is performed for dehydration and fixation, and after cooling, the dried tea leaves are crushed by a crushing device, sieved, and tea powder is obtained; the power of microwave treatment is 400 W, and the treatment time is 50 s; the particle size after sieving is 40-60 meshes;

[0077] (2) Three-stage extraction: the crushed tea leaves are taken, ethanol solution is added, and the first-stage extraction is performed to obtain the first-stage extract; the mixture after the first-stage extraction is subjected to the second-stage extraction to obtain the second-stage extract; the mixture after the second-stage extraction is subjected to the third-stage extraction to obtain the extract; the mass concentration of the ethanol solution is 50%, the weight ratio of the enzymatic hydrolysate to the ethanol solution is 1:4, the first-stage extraction temperature is 30°C, and the time is 40 min, the second-stage extraction temperature is 60°C, and the time is 40 min, and the third-stage extraction temperature is 80°C, and the time is 20 min;

[0078] (3) Extract treatment: the extract after the multi-stage extraction is subjected to centrifugation and filtration treatment to further purify the effective components in the extract, and tea crude extract is obtained; the tea crude extract is subjected to adsorption and desorption treatment in sequence by using a macroporous resin, and the extract after desorption is subjected to vacuum distillation treatment to obtain a tea mixture;

[0079] (4) Spray drying: the tea mixture is subjected to spray drying treatment to obtain the tea extract in powder form; the parameters of spray drying are as follows: the inlet air temperature is 100°C, the outlet air temperature is 80°C, and the feeding speed is 15 mL / min.

[0080] Comparative Example 8

[0081] Comparative Example 8 differs from Example 1 in that the four ingredients in the composition: ceramide, enzymatic tea extract, DNA sodium, and camellia japonica extract are directly mixed by stirring at a rotation speed of 500 r / min for 20 min to obtain the composition.

[0082] Test Example 1: Test of Anti-aging Repair Composition for Reducing Inflammatory Factors

[0083] I. Reagent Preparation

[0084] 1. Culture medium preparation: DMEM culture medium containing 10% (V / V) fetal bovine serum (FBS) is prepared.

[0085] 2. Trypsin-EDTA solution: 0.25% trypsin-EDTA solution is divided into small volumes (such as 5-10 mL), and is stored in a -20°C refrigerator. Before use, it is quickly thawed in a 37°C water bath, and repeated freezing and thawing is avoided to affect the activity.

[0086] 3. Lipopolysaccharide (LPS): LPS was prepared into a 1 mg / mL stock solution with sterile PBS and stored at -20°C. Before the experiment, the stock solution was diluted with culture medium to a working concentration of 1 μg / mL.

[0087] 4. Composition sample solution: The compositions of Examples 1-7 and Comparative Examples 1-8 were dissolved in DMSO to prepare a 10 mg / mL stock solution, which was stored at -20°C. When used, the stock solution was diluted with culture medium to a working concentration of 100 μg / mL, ensuring that the final concentration of DMSO did not exceed 0.1% to exclude its potential effects on cells.

[0088] 5. Dexamethasone: Dexamethasone was prepared into a 10 mM stock solution with anhydrous ethanol and stored at -20°C. Before the experiment, the stock solution was diluted with culture medium to a working concentration of 1 μM.

[0089] 6. ELISA kit-related reagents: According to the instructions of the selected ELISA kit for inflammatory factors such as TNF-α and IL-1α, prepare the required washing buffer, detection antibody, standard, and other reagents, and restore them to room temperature in advance.

[0090] II. Preparation of consumables

[0091] 1. Cell culture consumables: Prepare T75 culture flasks, 96-well cell culture plates, 15 mL and 50 mL centrifuge tubes, pipette tips (10 μL, 200 μL, 1000 μL), cell scrapers, etc., ensuring that all consumables are sterile products and are used as soon as possible after opening.

[0092] 2. Other consumables: Prepare 96-well plate sealing membranes, tin foil paper (for light-proof storage of reagents), ice boxes (for maintaining sample low temperature during operation), etc.

[0093] III. Preparation of instruments

[0094] 1. Cell culture-related instruments: Turn on the carbon dioxide incubator in advance, adjust the temperature to 37°C, the CO2 concentration to 5%, and the humidity to more than 95%, and perform cleaning and disinfection treatment. The centrifuge is set to 4°C for pre-cooling for cell centrifugation operation.

[0095] 2. Detection instruments: Preheat the enzyme marker according to the instructions, calibrate and perform performance testing to ensure the accuracy of the detection results. At the same time, prepare the vortex shaker, micropipette, etc., and perform calibration and function check.

[0096] IV. Cell culture

[0097] RAW264.7 mouse macrophage cells were inoculated in T75 culture bottles and cultured in a 37℃, 5% CO2 environment with DMEM medium containing 10% FBS. When the cells grew to the logarithmic growth phase, the cells were digested with 0.25% trypsin-EDTA, the cell density was adjusted to 2x10 5 cells / mL, and 100 μL of the cell suspension was added to each well of a 96-well plate.

[0098] Five, drug treatment and inflammation induction

[0099] (I) Group setting

[0100] Blank control group: 100 μL of cell suspension and 100 μL of medium were added to each well, without adding LPS and composition sample solution.

[0101] LPS group: 100 μL of cell suspension was added to each well, 50 μL of medium was added first, and then 50 μL of 1 μg / mL LPS solution was added to induce inflammation.

[0102] Sample group: 100 μL of cell suspension was added to each well, and 50 μL of 50 μg / mL composition sample solution of Examples 1-7 and Comparative Examples 1-8 was added (per well), and the 96-well plate was placed in a culture incubator for pre-treatment for 1 hour. After the pre-treatment was completed, 50 μL of 1 μg / mL LPS solution was added to each well.

[0103] Positive control group: 100 μL of cell suspension was added to each well, and 50 μL of 1 μM dexamethasone solution was added, and after pre-treatment for 1 hour, 50 μL of 1 μg / mL LPS solution was added.

[0104] (II) Specific operation

[0105] According to the group setting, the corresponding drugs and LPS solutions were accurately added using a multichannel pipette, and the final volume in each well was 200 μL. When adding the solution, try to avoid direct impact of the liquid on the cells, and slowly add the pipette tip to the edge of the well wall.

[0106] After the sample was added, the 96-well plate was gently shaken to mix the solution evenly, and the 96-well plate was sealed with a sealing film to prevent water evaporation and contamination. The treated cells were placed in a 37℃, 5% CO2 incubator for 24 hours. During the culture process, the incubator door was not frequently opened to maintain a stable culture environment.

[0107] Four, inflammation factor detection and result calculation

[0108] After 24 hours of culture, the 96-well plate was taken out of the incubator and placed on an ice box for operation to prevent degradation of inflammatory factors. The cell culture supernatant in the 96-well plate was transferred to a new 96-well plate, and the cell precipitate was avoided to be sucked as much as possible. If further treatment of the supernatant is needed (such as centrifugation to remove cell debris, etc.), centrifugation at 1000 rpm for 5 minutes at 4°C can be performed, and the supernatant is taken for detection. According to the operation instruction of the ELISA kit, the ELISA plate was placed in the enzyme label instrument, and the corresponding detection wavelength was selected according to the TNF-α and IL-1α inflammatory factor in the kit, and the absorbance (OD value) of each well was measured. The standard concentration was taken as the X axis, and the OD value was taken as the Y axis, and the curve was fitted by four-parameter logistic regression (4-PL) or linear regression. The sample OD value was substituted into the standard curve equation to calculate the TNF-α or IL-1α concentration (pg / mL), and the specific formula was as follows:

[0109] TNF-α inhibition rate (%) = (1-LPS group TNF-α concentration / sample group TNF-α concentration) x 100%;

[0110] IL-1α inhibition rate (%) = (1-LPS group IL-1α concentration / sample group IL-1α concentration) x 100%;

[0111] Among them, the sample group includes the sample group, the blank control group and the positive control group, and the specific data are shown in Table 2.

[0112] Table 2 TNF-α inhibition rate and IL-1α inhibition rate of the composition of each group

[0113] Group / Performance TNF-α inhibition rate / % IL-1α inhibition rate / % Example 1 78.4 66.3 Example 2 76.3 64.5 Example 3 76.5 64.8 Example 4 72.2 62.2 Example 5 76.2 63.9 Example 6 69.4 61.2 Example 7 69.9 61.5 Comparative Example 1 29.7 24.5 Comparative Example 2 50.7 41.2 Comparative Example 3 57.9 49.2 Comparative Example 4 35.2 30.3 Comparative Example 5 39.9 31.7 Comparative Example 6 65.6 55.7 Comparative Example 7 63.0 54.1 Comparative Example 8 38.2 30.0 Blank Control Group 0.2 0.4 Positive Control Group 83.5 75.3

[0114] It can be known from Table 2 that the greater the TNF-α inhibition rate and IL-1α inhibition rate values, the better the anti-inflammatory performance of the composition.

[0115] It can be known from the data of Example 1 and Comparative Examples 1-5 that the TNF-α inhibition rate and IL-1α inhibition rate of Comparative Examples 1-5 are all decreased to different degrees compared with Example 1, which indicates that the Camellia nitidissima Chang extract contains flavonoids (quercetin, kaempferol), which reduces TNF-α transcription by inhibiting the NF-κB pathway, and is the core of anti-inflammatory. The tea releases small molecule EGCG through enzymatic hydrolysis process, enhances permeability, and directly inhibits IL-1α converting enzyme activity. Chitosan has the effects of antibacterial and anti-inflammatory, and reduces inflammatory symptoms. DNA sodium accelerates tissue repair and indirectly reduces inflammation triggering.

[0116] It can be known from the data of Example 1 and Comparative Examples 6 and 7 that the composition of the active substance wrapped by the liposome has a great influence on the anti-inflammatory effect of the composition.

[0117] From the combination of Example 1 and Comparative Example 7, it can be seen that the anti-inflammatory performance of Comparative Example 7 is significantly lower than that of Example 1, which is probably because the tea extract of Comparative Example 7 is not subjected to enzymatic hydrolysis, and the tea releases small molecules of EGCG through the enzymatic hydrolysis process, enhances the permeability, and directly inhibits the IL-1α converting enzyme activity.

[0118] From the combination of Example 1 and the data of Examples 1-7, it can be seen that when the weight ratio of the Camellia japonica extract, DNA sodium, lecithin, cholesterol, ceramide, enzymatically hydrolyzed tea extract, and chitosan is (1.5-2):(0.05-0.1):(4-5):(0.8-1):(0.05-0.1):(0.05-0.1):(0.4-0.5), the anti-inflammatory effect of the composition is at a relatively optimal level.

[0119] Test Example 2: Test of Composition Promoting Collagen Production

[0120] I. Preparation of Reagents

[0121] 1. Preparation of culture medium: DMEM culture medium containing 10% (V / V) fetal bovine serum (FBS) is prepared.

[0122] 2. Trypsin-EDTA solution: 0.25% trypsin-EDTA solution is aliquoted into small volumes and stored at -20°C. Before use, it is quickly thawed at 37°C water bath to avoid the influence of repeated freezing and thawing on activity.

[0123] 3. PBS buffer: PBS buffer with pH 7.2-7.4 is prepared, sterilized by autoclaving, and stored at 4°C for cell washing.

[0124] 4. Cell lysis solution: according to the experimental requirements, a suitable cell lysis solution (such as RIPA lysis solution containing protease inhibitors) is selected and prepared on site to ensure the lysis effect.

[0125] 5. Collagen detection kit: carefully read the kit instructions, and take the required reagents (such as standards, detection antibodies, color developing agents, etc.) out of the refrigerator in advance and restore to room temperature.

[0126] 6. Sample composition solution: the compositions of Examples 1-7 and Comparative Examples 1-8 are dissolved with DMSO to prepare 10 mg / mL stock solutions, which are stored at -20°C. During the experiment, the medium is used to dilute to a working concentration of 100 μg / mL, and the final concentration of DMSO is not more than 0.1%.

[0127] II. Preparation of Consumables

[0128] Cell culture consumables: prepare sterile T75 culture bottles, 96-well cell culture plates, 15 mL and 50 mL centrifuge tubes, pipette tips (10 μL, 200 μL, 1000 μL), cell scrapers, etc., and use them as soon as possible after opening.

[0129] Other consumables: Prepare 96-well plate sealing film, ice box (for keeping the sample at low temperature), vortex shaker, tin foil paper (to store reagents in the dark), etc.

[0130] III. Instrument preparation

[0131] 1. Cell culture related instruments: Turn on the carbon dioxide incubator in advance, adjust the temperature to 37°C, the CO2 concentration to 5%, and the humidity to more than 95%, and clean and disinfect. The centrifuge is set to 4°C for pre-cooling for cell centrifugation.

[0132] 2. Detection instrument: Preheat and calibrate the enzyme label instrument to ensure detection accuracy. At the same time, prepare micropipettors, vortex shakers, etc., and perform function checks and calibrations.

[0133] IV. Cell culture and inoculation

[0134] Take the frozen human fibroblasts from the liquid nitrogen tank and quickly place them in a 37°C water bath for rapid thawing, constantly shaking the frozen tube during the process. After thawing, inoculate the cell suspension into a T75 culture flask, and use DMEM medium containing 10% FBS to culture at 37°C in a 5% CO2 environment. When the cells grow to the logarithmic growth phase, digest the cells with 0.25% trypsin-EDTA, adjust the cell density to an appropriate level (such as 2 x 10 5 cells / mL), and inoculate into a 96-well plate, adding an appropriate amount of cell suspension to each well (100 μL per well).

[0135] V. Drug treatment

[0136] 1. Group setting

[0137] Blank control group: Add 100 μL of cell suspension and 100 μL of medium to each well without adding sample composition solution.

[0138] Sample group: Add 50 μL of sample composition solution of 100 μg / mL of Examples 1-7 and Comparative Examples 1-8 (per well), and then add 50 μL of medium to make the final volume of each well 200 μL.

[0139] 2. Specific operation

[0140] Use a multichannel pipettor to accurately add the corresponding sample solution and medium according to the group setting. When adding the solution, try to avoid liquid impact on the cells, and you can slowly add the pipettor gun head to the edge of the well wall.

[0141] After the addition is completed, gently shake the 96-well plate to mix the solution evenly, seal the 96-well plate with a sealing film to prevent water evaporation and contamination. Place the treated cells in a 37°C, 5% CO2 incubator for 48 hours. During the incubation process, avoid frequently opening the incubator door to maintain a stable incubation environment.

[0142] Six, sample collection and collagen detection

[0143] 1. Cell lysis

[0144] After the incubation is completed, take the 96-well plate out of the incubator and place it on an ice box for operation to prevent collagen degradation. Discard the culture medium and gently rinse the cells with 100 μL of PBS twice to remove residual culture medium. Add 50 μL of cell lysis solution to each well, place the 96-well plate on ice for lysis for 30 minutes, and gently shake the 96-well plate every 5-10 minutes during the lysis process to ensure that the cells are fully lysed. After lysis is completed, transfer the lysate to a 1.5 mL centrifuge tube, centrifuge at 12000 rpm at 4°C for 10 minutes, collect the supernatant, and avoid sucking the precipitate. Transfer the supernatant to a new centrifuge tube and store it at -80°C for later use or immediately proceed to collagen detection.

[0145] 2. Collagen detection and result calculation

[0146] According to the instructions of the collagen detection kit, place the ELISA plate in the enzyme label instrument, select the appropriate wavelength (according to the kit), and measure the absorbance (OD value) of each well. Use the standard concentration as the X-axis and the OD value as the Y-axis to fit the curve with four-parameter logistic regression (4-PL) or linear regression. Substitute the sample OD value into the standard curve equation to calculate the collagen concentration (pg / mL), and the specific formula is as follows:

[0147] Collagen concentration increase rate (%) = (collagen concentration 样品组 -collagen concentration 空白对照组 ) / collagen concentration 空白对照组 × 100%; see Table 3.

[0148] Table 3: Collagen increase rate of samples in each group

[0149]

[0150]

[0151] As can be seen from Table 3, the higher the collagen promotion rate, the better the repair effect of the composition.

[0152] From the data of Example 1 and Comparative Examples 1-5, it can be seen that the collagen promotion rates of Comparative Examples 1-5 are all decreased to different degrees compared with Example 1, which shows that DNA sodium as a direct activator of collagen synthesis significantly improves the dermal layer density by stimulating fibroblast proliferation and type III collagen expression. The Camellia japonica extract rich in flavonoids (such as kaempferol) protects the collagen structure by inhibiting MMP-1 enzyme, and synergistically enhances the collagen density with DNA sodium. The enzymatic tea extract releases small molecules EGCG (epigallocatechin gallate) after enzymolysis, which penetrates the dermal layer to activate collagen gene transcription. Ceramide reduces inflammatory damage by strengthening the skin barrier and protecting fibroblast function, thereby indirectly promoting collagen synthesis.

[0153] From the data of Example 1 and Comparative Examples 6 and 7, it can be seen that the collagen promotion effect of the liposome-encapsulated ingredient combination has a greater impact.

[0154] From the data of Example 1 and Comparative Example 7, it can be seen that the anti-inflammatory performance of Comparative Example 7 is significantly decreased compared with Example 1, which may be because the tea extract of Comparative Example 7 is not enzymolysed, and the content of EGCG is reduced.

[0155] From the data of Example 1 and Examples 1-7, it can be seen that when the weight ratio of Camellia japonica extract and DNA sodium is (1.5-2):(0.05-0.1), and the weight ratio of lecithin, cholesterol, ceramide, enzymatic tea extract and chitosan is (4-5):(0.8-1):(0.05-0.1):(0.05-0.1):(0.4-0.5), the effect of the composition on promoting collagen production is at a relatively optimal level.

[0156] Application Examples 1-7 and Comparative Application Examples 1-8

[0157] The compositions of Examples 1-7 and Comparative Examples 1-8 are added to the emulsion at a concentration of 10 wt%, to obtain the emulsions of Application Examples 1-7 and Comparative Application Examples 1-8, and the formulations are shown in Table 4.

[0158] The preparation method of the emulsions of Application Examples 1-7 and Comparative Application Examples 1-8 comprises the following steps:

[0159] (1) Mix the thickening agent and the humectant uniformly, add pure water, heat at a water bath temperature of 75°C for 10 min, then homogenize uniformly at 4000 rpm for 5 min, and store for use after keeping warm, to obtain Component A.

[0160] (2) Mix the oil and the emulsifier, heat to 75°C, and stir to dissolve uniformly, and store for use after keeping warm, to obtain Component B.

[0161] (3) The A component is heated to 75°C, the B component is added, and homogenized at 4000 rpm for 5 min to obtain an emulsified base.

[0162] (4) The emulsified base is stirred and cooled to 50°C, the anti-aging protective composition is added, and stirring is continued until the material is uniform. Then, the preservative and pH adjuster are added to adjust the pH to 6.5, the stirring is stopped, the material is discharged, and an emulsion is obtained.

[0163] Table 3 Emulsion formulations of application examples 1-7 and comparative application examples 1-7

[0164]

[0165] Comparative application example 9

[0166] The emulsion of comparative application example 9 is prepared without adding the anti-aging protective composition, and an equal amount of deionized water is used instead of the composition. The preparation method is the same as that of application example 1.

[0167] Test example 3: human patch test of emulsion

[0168] Thirty volunteers, 15 males and 15 females, aged 20-50 years, are recruited, and a closed patch test method is used. Equal amounts (0.5g-0.6g) of test samples (emulsions prepared in application examples 1-7 and comparative application examples 1-9) are placed in a specific patch tester, which is attached to the volunteer's arm with low-sensitivity adhesive tape. Eight groups of test samples are attached to each arm, and the skin is lightly pressed to ensure uniform attachment. The patch tester is left on the skin for 24 hours. After 24 hours, the patch tester is removed, and the skin reaction is observed at 0.5h, 24h, and 48h. The results are recorded. The adverse reaction grade of the skin is shown in Table 4 below.

[0169] Table 4 Adverse reaction grade of the skin

[0170]

[0171] After testing, the emulsions provided in application examples 1-7 and comparative application examples 1-9 are all negative reactions after human patch testing, and are safe and non-irritating to human skin.

[0172] Test example 4: human anti-wrinkle firming and soothing efficacy test

[0173] 1. Experimental basis

[0174] The human efficacy evaluation test method in the "Cosmetic Safety Technical Specification" (2015 edition) is referred to.

[0175] 2. Subject selection

[0176] Inclusion criteria: 45-60 year old Asian adult female volunteers are recruited, who meet the following conditions:

[0177] a) Clinical evaluation of skin laxity (grade I-III according to clinical grading criteria);

[0178] b) Self-reported history of skin sensitivity (confirmed by dermatologist evaluation);

[0179] c) No history of serious systemic disease or skin disease.

[0180] Exclusion criteria: pregnant / lactating women, severe allergic constitution, participation in other clinical trials within the last 3 months.

[0181] Number of subjects: A total of 90 qualified volunteers were included, divided into 16 groups of 5 people each according to the random number table method.

[0182] 3. Sample application method

[0183] Test samples: Emulsions prepared in Application Examples 1-7 and Comparative Application Example 1-9 (double-blind number) were used, among which the serum prepared in Comparative Application Example 9 was the blank control group, and the rest of the application examples and comparative examples were sample groups.

[0184] Usage: The volunteers applied 1 mL of the sample evenly to the whole face after cleansing in the morning and evening every day, and gently massaged until complete absorption.

[0185] 4. Test period and process

[0186] Test period: Lasting for 14, 28 and 56 days (D0, D 14 , D 28 and D 56 ).

[0187] Visit time points: Instrument detection was performed at D0 (baseline period), D 14 , D 28 and D 56 (end of test).

[0188] Preparation before detection:

[0189] a) The subject used a unified non-irritating cleansing product to clean the face after visiting;

[0190] b) Rest for 30 minutes in a constant temperature and humidity environment (temperature 21±1℃, humidity 50±10%);

[0191] c) Keep eyes closed and relaxed during detection, avoid facial expression interference.

[0192] The facial skin condition on the 14th day, 28th day and 56th day after use of the test product was observed, so as to comprehensively evaluate the firming and anti-wrinkle effect of the test product. The specific test parameters and equipment in the experiment are shown in Table 5, and the evaluation parameters are shown in Table 6.

[0193] Result determination: after the product is used, the values of the test area indicators R2 / R7 / Q1, WrinkleArea, and WrinkleLength increase and have significant differences, indicating that the test sample has the effect of firming and anti-wrinkling; on the contrary, if the parameters do not increase and do not have significant differences, it indicates that the test product does not have the effect of firming and anti-wrinkling.

[0194] Table 5 Test parameters and equipment

[0195]

[0196] Table 6 Evaluation parameters

[0197]

[0198]

[0199] The evaluation parameter calculation formula is as follows:

[0200] (T14, 28, 56) R2 value improvement rate = (R2 T0 -R2 Tn ) / R2 T0 ×100%; wherein, T n =T 14 , T 28 , T 56 .

[0201] (T14, 28, 56) WrinkleArea value improvement rate = (WrinkleArea T0 -WrinkleArea Tn ) / WrinkleArea T0 ×100%; wherein, T n =T 14 , T 28 , T 56 .

[0202] (T14, 28, 56) Wrinkle Length value improvement rate = (Wrinkle Length T0 -Wrinkle Length Tn ) / Wrinkle Length T0 ×100%; wherein, T n =T 14 , T 28 , T 56 . Data are shown in Table 7.

[0203] Table 7 Test results after the emulsion of each group is used

[0204]

[0205]

[0206] From Table 7, it can be seen that, in combination with the data of application example 1 and comparative application examples 1-5, the liposome mainly composed of camellia sinensis leaf extract, DNA sodium, ceramide, enzyme-decomposed tea leaf extract and chitosan can synergistically improve the anti-wrinkle and elasticity-improving effects of the emulsion.

[0207] In combination with the data of application example 1 and comparative application example 6, it can be seen that the performance of the emulsion is significantly lower than that of application example 1, which may be because the anti-aging effect of calendula extract is not as good as that of camellia sinensis leaf extract.

[0208] In combination with the data of application example 1 and comparative application example 7, it can be seen that the performance of the emulsion is lower than that of application example 1, which may be because the enzyme-decomposed tea leaf extract decomposes the cell wall of tea leaf accurately, releases more tea polyphenols, and makes the active ingredients in the tea leaf extract more easily absorbed by the skin to play the effects of antioxidant and anti-inflammatory.

[0209] In combination with the data of application example 1 and comparative application example 8, it can be seen that, in comparative application example 8, the raw materials of the composition are directly mixed without using liposomes as the carrier of the components, and the performance of the emulsion is significantly lower than that of application example 1, which may be because the form of liposomes can promote the transdermal absorption of active ingredients, and the use of coating liposome wrapping technology to wrap active substances can better help active substances penetrate the skin to achieve significant anti-aging and protection effects.

[0210] In summary, the anti-wrinkle, protection and elasticity-improving effects of the emulsion are mainly based on the addition of the anti-aging and protection composition, such as Figure 1 As shown in Table 8, after using the emulsion of application example 1 for 56 days, the crow's feet, under-eye wrinkles and marionette lines are all faded to different degrees, which may be because the anti-aging and protection composition of the application can promote collagen synthesis, enhance skin elasticity, and play multiple dimensions such as antioxidant and repair of damaged skin barrier, to comprehensively improve the aging condition of the skin.

[0211] Finally, it should be noted that the above examples are only used to illustrate the technical solutions of the application and not to limit the protection scope of the application. Although the application has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the application can be modified or replaced by equivalents without departing from the essence and scope of the technical solutions of the application.

Claims

1. An anti-aging and repairing composition containing Camellia chrysantha extract, characterized in that: The liposome composition comprises, from the inside to the outside, a core layer, a shell layer and a chitosan coating; wherein the core layer comprises a camellia chrysantha extract and sodium DNA; the shell layer comprises lecithin, cholesterol, ceramide and enzymatically hydrolyzed tea extract; the chitosan coating comprises chitosan; and the weight ratio of the camellia chrysantha extract, sodium DNA, lecithin, cholesterol, ceramide, enzymatically hydrolyzed tea extract and chitosan is (0.1-10): (0.001-2): (0.1-10): (0.1-5): (0.001-2): (0.001-2): (0.01-3).

2. The anti-aging and repairing composition containing the Camellia chrysantha extract according to claim 1, wherein The weight ratio of the camellia chrysantha extract to sodium DNA is (1.0-3.0): (0.01-0.5).

3. The anti-aging and repairing composition containing the Camellia chrysantha extract according to claim 1, wherein: The weight ratio of the lecithin, cholesterol, ceramide, enzymatically hydrolyzed tea extract and chitosan is (1-8): (0.5-3): (0.01-1): (0.01-0.5): (0.1-1).

4. The anti-aging and repairing composition containing the Camellia chrysantha extract according to claim 1, wherein: The weight ratio of the camellia chrysantha extract to DNA sodium is (1.5-2): (0.05-0.1).

5. The anti-aging and repairing composition containing the Camellia chrysantha extract according to claim 1, wherein The weight ratio of the lecithin, cholesterol, ceramide, enzymatically hydrolyzed tea extract and chitosan is (4-5): (0.8-1): (0.05-0.1): (0.05-0.1): (0.4-0.5).

6. The anti-aging and repairing composition containing the Camellia chrysantha extract according to claim 1, wherein: The method for preparing the enzymatically hydrolyzed tea extract comprises the following steps: (1) Raw material pretreatment: fresh, clean, and non-rotten tea leaves are selected as raw materials, and microwave treatment is performed to dehydrate and fix the tea leaves. After cooling, the dried tea leaves are crushed using a crushing device and sieved to obtain tea powder. The microwave treatment power is 300-500W, the treatment time is 40-60s, and the sieved particle size is 40-60 mesh. (2) Enzymatic hydrolysis: adding sterile deionized water to the tea powder, mixing and stirring uniformly, adding a complex enzyme, performing enzymatic hydrolysis at a temperature of 35-45°C for 2-5 hours, and performing enzyme inactivation treatment on the enzymatically hydrolyzed mixture to obtain an enzymatic hydrolyzate; wherein the amount of sterile deionized water added is 6-7 times the weight of the tea powder, the complex enzyme includes cellulase and pectinase in a weight ratio of 1:1, and the amount of the complex enzyme added is 0.6%-1% of the weight of the tea powder; (3) Three-stage extraction: adding ethanol solution to the enzymatic hydrolysate, performing the first stage extraction, and obtaining the first stage extract; performing the second stage extraction on the mixed solution after the first stage extraction, and obtaining the second stage extract; performing the third stage extraction on the mixed solution after the second stage extraction, and obtaining the extract; wherein the mass concentration of the ethanol solution is 45-55%, the weight ratio of the enzymatic hydrolysate to the ethanol solution is 1:(3-5), the first stage extraction temperature is 25-35°C, the time is 35-45 min, the second stage extraction temperature is 55-65°C, the time is 35-45 min, and the third stage extraction temperature is 75-85°C, the time is 15-25 min; (4) Extraction liquid treatment: The extract after the multi-stage extraction is centrifuged and filtered to obtain a crude tea extract, and the crude tea extract is sequentially subjected to adsorption and desorption treatment using a macroporous resin. The desorbed extract is vacuum distilled to obtain a tea mixture; (5) Spray drying: The tea mixture is spray dried to obtain the enzymatically hydrolyzed tea extract in powder form.

7. The method for preparing the anti-aging and repairing composition containing the Camellia chrysantha extract according to any one of claims 1 to 6, characterized in that: The following steps are involved: S1. Add lecithin, cholesterol, ceramide, enzymatically hydrolyzed tea extract, and anhydrous ethanol to a round-bottom flask, install the round-bottom flask on a rotary evaporator, start stirring and heat in a water bath until the components are dissolved to form a lipid solution; evaporate the anhydrous ethanol in the lipid solution until the lipid solution forms a thin film on the inner wall of the flask; the amount of anhydrous ethanol added is 2-3 times the mass of lecithin, cholesterol, ceramide, and enzymatically hydrolyzed tea extract; S2. Dissolve the Camellia chrysantha extract and sodium DNA in water to obtain a nuclear layer solution, add the nuclear layer solution to the round-bottom flask in which the thin film has been formed in step S1, start stirring and heat in a water bath until the lipid film is completely hydrated to form multilamellar liposomes, and then ultrasonically treat the multilamellar liposomes to obtain a liposome suspension; wherein the mass concentration of the nuclear layer solution is 30-40%, the stirring speed is 500-600 r / min, and the temperature of the water bath is 35-45°C; S3. Chitosan coating: Dissolve chitosan in 0.1%-1% glacial acetic acid solution to prepare a chitosan solution with a mass concentration of 0.2%-1.0%. Mix the liposome suspension with the prepared chitosan solution at a ratio of 1:0.

5. Stir at 25-30°C and let stand to stabilize electrostatic binding. Centrifuge to obtain chitosan-coated liposomes. S4. Ultrafiltration: Ultrafiltration is performed on the chitosan-coated liposomes to remove unencapsulated components and free lipids, thereby obtaining the anti-aging and repairing composition containing the Camellia chrysantha extract.

8. Use of the anti-aging and repairing composition containing the Camellia chrysantha extract according to any one of claims 1 to 6 in the preparation of cosmetics.

9. An emulsion, characterized in that The invention comprises the following raw materials in weight percentage: 5%-20% of the anti-aging repair composition containing the camellia chrysantha extract according to any one of claims 1 to 6, 0.05%-0.5% of a thickener, 1%-10% of a moisturizer, 1%-6% of an emulsifier, 5%-15% of oil, 0.5%-3% of a preservative and 0.01%-0.3% of a pH regulator, and the balance is deionized water.

10. The method for preparing the emulsion according to claim 9, characterized in that: The following steps are involved: (1) Mix the thickener and the moisturizer evenly, add deionized water, heat to 70-80°C, homogenize, and keep warm for later use to obtain component A; (2) Mix the oil and emulsifier, heat to 70-80°C, stir to dissolve, and keep warm for later use to obtain component B; (3) Heat component A to 70-80°C, add component B, and homogenize to obtain an emulsified base. (4) The emulsified base is stirred and cooled to 30-50° C., the anti-aging repair composition containing the Camellia chrysantha extract is added, and stirring is continued until the material is uniform, and then a preservative and a pH adjuster are added to adjust the pH to 6.0-6.5, and stirring is stopped. The material is discharged to obtain the emulsion.

Images