Anti-aging repairing composition for sensitive skin as well as preparation method and application of anti-aging repairing composition

Abstract

The invention discloses an anti-aging repairing composition for sensitive skin as well as a preparation method and application of the anti-aging repairing composition, and belongs to the technical field of daily cosmetic raw material preparation. The anti-aging repair composition is prepared by compounding the following components in parts by mass: 0.01 to 1 part of fisetin, 0.01 to 1.5 parts of acetyl hexapeptide-1, 0.01 to 0.3 part of tuberose polysaccharide and 1 to 4 parts of European chestnut leaf extract. Meanwhile, the invention optimizes a green preparation process combining dynamic countercurrent extraction and macroporous resin column purification of fisetin. The four components of the composition have a synergistic effect, a cell-blood vessel-nerve triple barrier repair anti-aging network is constructed, and the composition has excellent anti-oxidation, anti-inflammatory and anti-aging effects, is safe and non-irritant, perfectly meets the use requirements of sensitive skin, and can be widely applied to preparation of various anti-inflammatory, anti-oxidation and anti-aging skin care products.

Description

Technical Field

[0001] This invention belongs to the technical field of preparation of raw materials for daily cosmetics, specifically relating to an anti-aging and repairing composition for sensitive skin, its preparation method and application. Background Technology

[0002] Currently, fisetin extraction technology has significant limitations: traditional organic solvent methods (such as methanol / ethanol reflux), while efficient, inevitably introduce toxic solvent residues, posing a potential irritation risk to sensitive skin. Simultaneously, high-temperature processes easily lead to the degradation of heat-sensitive fisetin, while poor extraction selectivity results in numerous impurities and purification difficulties. It is difficult to simultaneously achieve high extraction efficiency, high product purity, no harmful residues, and preserved activity, failing to meet the stringent safety and quality standards for high-end sensitive skin cosmetic raw materials. Furthermore, industry solutions for anti-aging in sensitive skin mainly rely on high-concentration single ingredients or gentle plant-based complexes focused on soothing and repair. While the former can promote collagen regeneration, it can easily exacerbate skin sensitivity, while the latter has limited effectiveness due to a lack of core anti-aging mechanisms such as clearing aging cells, making it difficult to achieve both high efficiency and gentleness. Summary of the Invention

[0003] To address the shortcomings of existing technologies, the present invention aims to provide a compositional formulation system that simultaneously achieves barrier repair and precise anti-aging, breaking through the industry bottleneck of existing sensitive skin anti-aging products that "cannot balance efficacy and gentleness," and solving the problems that traditional anti-aging ingredients easily aggravate inflammatory reactions in sensitive skin, while gentle repair formulations lack core anti-aging mechanisms.

[0004] To achieve the above objectives, the present invention discloses the following technical solutions: In a first aspect, the present invention provides an anti-aging repair composition, which, by mass parts, comprises the following components: 0.01-1 parts of fisetin, 0.01-1.5 parts of acetyl hexapeptide-1, 0.01-0.3 parts of tuberose polysaccharide, and 1-4 parts of European chestnut leaf extract.

[0005] Preferably, the composition comprises, by weight parts, the following components: 0.1-0.8 parts of fisetin, 0.1-0.8 parts of acetyl hexapeptide-1, 0.1-0.2 parts of tuberose polysaccharide, and 1.1-3 parts of European chestnut leaf extract.

[0006] More preferably, the composition comprises, by weight parts, the following components: 0.4-0.6 parts of fisetin, 0.4-0.6 parts of acetyl hexapeptide-1, 0.14-0.16 parts of tuberose polysaccharide, and 1.9-2.1 parts of European chestnut leaf extract.

[0007] More preferably, the method for preparing the fisetin includes the following steps: Step 1: Crush and sieve the dried sawdust from the wax tree to obtain wax tree powder for later use; Step 2: Dynamic countercurrent extraction of wax tree powder to obtain an extract containing the target compound; Step 3: Concentrate the extract containing the target compound to obtain a crude extract of fisetin. Step 4: Purify and separate the crude extract of fisetin using a macroporous adsorption resin column, and collect the target eluent. Step 5: Concentrate the target eluent under reduced pressure and recrystallize it at a lower temperature to obtain fisetin crystals; Step 6: Vacuum dry the fisetin crystals to obtain fisetin.

[0008] More preferably, in step 2, the dynamic countercurrent extraction specifically involves using a 60%-70% ethanol aqueous solution as a solvent and performing 3-5 stages of continuous countercurrent extraction at 60-75°C and a material-to-liquid ratio of 1:8-1:12.

[0009] More preferably, in step 3, the concentration specifically involves concentrating the extract under reduced pressure at a vacuum of -0.08 to -0.09 MPa and a temperature of 55-65°C, recovering the ethanol, and obtaining a crude extract.

[0010] More preferably, in step 4, the purification and separation specifically involves redissolving the crude extract in water at a mass-to-volume ratio of 1:3 to 1:5 g / mL, filtering to obtain a clear sample solution, and loading the clear solution onto a macroporous adsorption resin column at a flow rate of 1 to 2 BV / h, with a sample volume of 0.3 to 0.5 BV of the resin's wet volume. After loading, the column is first eluted with 3 to 5 BV of pure water at a flow rate of 2 to 3 BV / h to remove water-soluble impurities. After elution, the column is then eluted again with 3 to 5 BV of 70 v / v% to 80 v / v% ethanol aqueous solution at a flow rate of 1 to 2 BV / h, and the eluent rich in fisetin is collected.

[0011] More preferably, in step 5, the cooling recrystallization specifically involves concentrating the eluent under reduced pressure at 50-60°C to 1 / 5-1 / 10 of its original volume, then cooling it to 0-4°C via a programmed cooling process for recrystallization, followed by filtration to obtain crystals.

[0012] Secondly, the present invention provides the application of the composition described in the first aspect in the preparation of skin care products with anti-inflammatory, antioxidant and anti-aging effects.

[0013] Thirdly, the present invention provides a lotion that removes aging skin cells and improves the skin aging of sensitive skin, the lotion containing the composition described in the first aspect.

[0014] Preferably, the emulsion further contains at least one of a thickener, a humectant, a pH adjuster, an emulsifier, an oil, a preservative, and a chelating agent.

[0015] More preferably, the thickener is selected from at least one of xanthan gum, carbomer, hydroxyethyl acrylate / sodium acryloyldimethyl taurate copolymer, ammonium acryloyldimethyl taurate / VP copolymer, hydroxyethyl cellulose, and cetyl alcohol.

[0016] More preferably, the moisturizer is selected from at least one of allantoin, sodium polyacrylate, panthenol, β-glucan, trehalose, caprylyl glycol, dipropylene glycol, sodium hyaluronate, 1,2-butanediol, glycerin, and tremella polysaccharide.

[0017] More preferably, the pH adjuster is selected from at least one of arginine, citric acid, and NaOH.

[0018] More preferably, the emulsifier is selected from 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, cetyl phosphate potassium, and sodium stearoyl glutamate.

[0019] More preferably, the oil is selected from at least one of caprylic / capric triglyceride, polydimethylsiloxane, jojoba oil, grape seed oil, meadowfoam seed oil, squalane, macadamia nut oil, and camellia oil.

[0020] More preferably, the preservative is selected from at least one of 1,2-hexanediol, 1,2-pentanediol, ethylhexylglycerin, p-hydroxyacetophenone, phenoxyethanol, octanoyl hydroxamic acid, and sodium benzoate.

[0021] The beneficial effects of this invention are: 1. This invention is the first to scientifically combine fisetin, which can specifically eliminate aging cells, European chestnut leaf extract, which strengthens the microvascular barrier, tuberose polysaccharide, which repairs the physical barrier, and acetyl hexapeptide-1, which regulates nerve signals, to construct a synergistic repair and anti-aging network that targets the cellular, vascular, and nerve barriers simultaneously. This design breaks through the bottleneck of traditional anti-aging formulas that are difficult to balance irritation and efficacy, and pioneers a new path of "using barrier repair to empower gentle anti-aging". 2. This invention employs a dynamic countercurrent extraction combined with macroporous resin column purification process to prepare fisetin, using wood wax tree, a byproduct of food processing, as raw material. This provides a wide availability of raw materials at low cost, and achieves high-value utilization of the byproduct, making it environmentally friendly. By precisely optimizing the extraction solvent, temperature, and purification method, fisetin degradation caused by high temperatures during traditional organic solvent thermal reflux extraction is avoided, while significantly reducing the risk of organic solvent residue. Detailed Implementation

[0022] To better illustrate the objectives, technical solutions, and advantages of this invention, the invention will be further described below with reference to specific embodiments. Those skilled in the art should understand that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

[0023] Unless otherwise specified, the experimental methods used in the specific embodiments are all conventional methods; the materials and reagents used are all commercially available unless otherwise specified; and the percentages mentioned in the specific embodiments are all mass percentages unless otherwise specified.

[0024] I. In this invention Acetyl hexapeptide-1: Purchased from Nanjing Genscript Biotech Co., Ltd., product code: RP30144CN-100; Tuberose polysaccharide: Purchased from Xinyang Mufan Biotechnology Co., Ltd., product code: 11988; European chestnut leaf extract: purchased from Henan Tianfu Chemical Co., Ltd., product code: European chestnut leaf extract; Wax tree: Obtained from Jiaoling County Futai Biotechnology Co., Ltd., product name: Lacquer tree.

[0025] II. Preparation of Fesetone 1. Dynamic reverse flow extraction The specific steps are as follows: Step 1, Raw material pretreatment: Crush the dried sawdust from the wood wax tree and pass it through a 40-mesh sieve for later use; Step 2, Extraction: The raw material is put into a countercurrent extraction device, and 70 v / v% ethanol aqueous solution is used as solvent. Three-stage continuous countercurrent extraction is carried out at 65℃ and a material-to-liquid ratio of 1:10. Step 3, Concentration: The extract is concentrated under reduced pressure at a vacuum of -0.09 MPa and a temperature of 60°C to recover ethanol and obtain crude extract. Step 4, Purification: The crude extract was redissolved in water at a mass-to-volume ratio of 1:5 g / mL, filtered, and the resulting clear solution was loaded onto an AB-8 macroporous adsorption resin column at a flow rate of 1.5 BV / h. The loading volume was 0.4 BV of the resin wet volume. After loading, the column was eluted with 5 BV of pure water at a flow rate of 2.5 BV / h to remove water-soluble impurities. After elution, the column was eluted with 5 BV of 75 v / v% ethanol aqueous solution at a flow rate of 1.2 BV / h, and the eluent rich in fisetin was collected. Step 5, Purification: The eluent is concentrated to 1 / 7 of its original volume under reduced pressure at a vacuum of -0.09 MPa and a temperature of 60°C. It is then cooled to 2°C and recrystallized, filtered, to obtain crystals. Step 6, Drying: The obtained crystals were vacuum dried at a vacuum degree of -0.095 MPa and a temperature of 45 °C to obtain fisetin ①.

[0026] 2. Organic solvent hot reflux extraction method The specific steps are as follows: Step 1, Raw material pretreatment: Crush the dried sawdust of wax tree into powder through a 40-mesh sieve to obtain wax tree powder for later use; Step 2, Extraction: Add 70 v / v% ethanol aqueous solution to the wood wax powder at a material-to-liquid ratio of 1:20 g / mL, heat and reflux at 80℃ for 2 hours, filter while hot, collect the filtrate, repeat the extraction once with the filter residue, combine the two filtrates, concentrate under reduced pressure at 60℃ and vacuum degree -0.09MPa to recover ethanol, and obtain a thick extract. Step 3, Purification: Disperse the thick extract in water, extract three times with an equal volume of ethyl acetate, combine the organic phases, dry with anhydrous sodium sulfate, and concentrate under reduced pressure to dryness to obtain fisetin ②.

[0027] III. An anti-aging repair composition 1. Composition 1-6 The anti-aging repair composition was obtained by compounding according to the mass proportions in Table 1. Table 1. Mass parts of each component in the composition Raw material name Composition 1 Composition 2 Composition 3 Composition 4 Composition 5 Composition 6 Fesetone① 0.01 0.1 0.4 0.6 0.8 1 Acetyl hexapeptide-1 0.01 0.1 0.4 0.6 0.8 1.5 Tuberose polysaccharides 0.01 0.1 0.14 0.16 0.2 0.3 European chestnut leaf extract 1 1.4 1.9 2.1 3 4 2. Composition 7-10 Composition 7-10 is an adjustment based on the formulation of composition 3, specifically compounded according to the mass parts in Table 2 to obtain composition 7-10; Table 2. Mass parts of each component in the composition Raw material name Composition 7 Composition 8 Composition 9 Composition 10 Fesetone① / 0.4 0.4 0.4 Acetyl hexapeptide-1 0.4 / 0.4 0.4 Tuberose polysaccharides 0.14 0.14 / 0.14 European chestnut leaf extract 1.9 1.9 1.9 / Note: " / " in the table indicates no addition.

[0028] 3. Composition 11 Composition 11 is based on the formulation of composition 3, but with fisetin ① replaced by fisetin ②. The specific compounding is carried out according to the mass parts in Table 3 to obtain composition 11. Table 3. Mass parts of each component in the composition Raw material name Composition 11 Fesetone ② 0.4 Acetyl hexapeptide-1 0.4 Tuberose polysaccharides 0.14 European chestnut leaf extract 1.9 IV. Testing of Fesetone Purity and Composition Properties 1. Determination of fisetin purity by HPLC external standard method 1.1 Instruments and Reagents Instruments: High performance liquid chromatograph (equipped with ultraviolet detector or DAD), analytical balance, ultrasonic instrument, 0.45μm microporous filter membrane.

[0029] Reagents: Fesedon reference standard (purity ≥98%), acetonitrile (chromatographic grade), phosphoric acid (analytical grade or chromatographic grade), ultrapure water, methanol (chromatographic grade).

[0030] 1.2 Chromatographic conditions Chromatographic column: C18 column (4.6mm×250mm, 5μm).

[0031] Mobile phase: acetonitrile: 0.1% phosphoric acid aqueous solution, gradient elution; flow rate: 1.0 mL / min; column temperature: 30°C; detection wavelength: 360 nm; injection volume: 10 μL.

[0032] 1.3 Solution Preparation Reference stock solution: Accurately weigh about 10 mg of fisetin reference standard, place it in a 50 mL brown volumetric flask, dissolve and dilute to the mark with methanol, shake well to obtain a stock solution with a concentration of about 0.2 mg / mL.

[0033] Series of reference solutions: Accurately measure an appropriate amount of the above stock solution, dilute it with methanol, and prepare a series of standard solutions with concentrations of approximately 5, 10, 20, 40, and 60 μg / mL.

[0034] Test solution: Accurately weigh approximately 5 mg each of your samples fisetin ① and fisetin ②, place them in a 25 mL brown volumetric flask, dissolve and dilute to the mark with methanol by sonication, and shake well. Filter through a 0.45 μm microporous membrane and collect the filtrate for later use.

[0035] 1.4. Determination Method System suitability: Take a standard solution of appropriate concentration (e.g., 20 μg / mL), inject it 5-6 times consecutively, and record the retention time and peak area of ​​fisetin. The RSD of retention time should be ≤1.0%, and the RSD of peak area should be ≤2.0%.

[0036] Standard curve construction: Inject a series of reference solutions sequentially. Plot concentration (C, μg / mL) on the x-axis and peak area (A) on the y-axis, and perform linear regression to obtain the standard curve equation A = kC + b. The correlation coefficient r is required. 2 ≥0.999.

[0037] Sample determination: Inject the test solutions of fisetin ① and fisetin ② separately, and record the peak areas of the two fisetins (A_sample).

[0038] Calculate purity: Substitute sample A into the standard curve equation to calculate the concentration of the two fisetinones in the test solution (sample C, μg / mL).

[0039] Calculate sample purity using the following formula: Purity (%) = (C_sample × V × 10) -3 ) / m×100% C_sample: Concentration calculated from the standard curve (μg / mL); V: Volume of the test solution (mL); m: Mass of the sample taken (mg); 10 -3 : The coefficient for converting μg to mg The results are shown in Table 4: Table 4 Fesetone① Fesetone ② purity(%) 98.8 84.6 2. In vitro detection of the inhibition rate of aging-related secretory phenotype (IL-6) Principle: Senescent cells secrete large amounts of pro-inflammatory cytokines (IL-6). If the composition can inhibit senescent cells, the concentration of these pro-inflammatory cytokines in their culture supernatant will decrease. The inhibition rate can be calculated by comparing the concentration difference between the treatment group and the model group.

[0040] 2.1 Constructing a model of senescent cells Cell selection: Human dermal fibroblasts (HDFs) were used.

[0041] Induced senescence: Cells were treated with a sublethal concentration of hydrogen peroxide (H2O2) (150-200 μM) for 24 h. After induction, the cells should stop proliferating and exhibit senescent morphology.

[0042] Model validation: SA-β-Gal staining (senescent cells appear blue) was used to confirm that the proportion of senescent cells in the model exceeded 70%, ensuring the model's success.

[0043] 2.2 Experimental Grouping: Blank control group: young cells that have not been induced to age.

[0044] Model control group: No drug treatment was given after induced aging.

[0045] Experimental group: After induced aging, composition 1-11 (0.1%) was added.

[0046] Procedure: Add the drug to the cell culture medium and continue culturing for 24-48 hours. After the culture is complete, collect the cell culture supernatant, centrifuge at 3000 rpm for 10 minutes, and use the supernatant for later use.

[0047] 2.3 ELISA Detection Follow the operating procedures on the ELISA kit. Finally, measure the absorbance (OD value) of each well at a wavelength of 450 nm. Plot a standard curve based on the concentration and OD value of the standard. Substitute the OD values ​​of the experimental group and the model group into the standard curve to calculate the actual concentration (pg / mL) of IL-6 in the supernatant, and calculate the inhibition rate. The formula for calculating the inhibition rate is as follows: Inhibition rate (%) = [1 - (concentration in experimental group / concentration in model group)] × 100% The results are shown in Table 5.

[0048] 3. In vitro anti-inflammatory effect assay (TNF-α) 3.1 Cell Culture RAW264.7 mouse macrophages were seeded in T75 culture flasks and cultured in DMEM medium containing 10% FBS at 37°C and 5 v / v % CO2. When the cells reached the logarithmic growth phase, they were digested with 0.25% trypsin-EDTA to adjust the cell density to 2 × 10⁶ cells / year. 5 Cells / mL were seeded into 96-well plates, with 100 μL of cell suspension added to each well.

[0049] 3.2 Preparation of the test sample Take each composition 1-11 and dilute it with DMEM medium containing 10% FBS to prepare a test sample 1-11 with a concentration of 100 μg / mL.

[0050] 3.3 Drug treatment and inflammation induction Group settings: Blank control group: 100 μL of culture medium.

[0051] LPS group: 50 μL culture medium + 50 μL 1 μg / mL LPS to induce inflammatory response.

[0052] Sample group: 50 μL of 100 μg / mL test sample 1-11, pretreated for 1 h, and then 50 μL of 1 μg / mL LPS was added.

[0053] 3.4. Specific operations In a 96-well plate, each well was first seeded with 100 μL of cell suspension (2 × 10⁶ cells / well). 5 After culturing for 24 hours and allowing the cells to adhere, the culture medium was changed and samples were added according to the group settings (ensuring a final volume of 200 μL for each group). The treated cells were then placed in a 37°C, 5v / v%CO2 incubator for 24 hours.

[0054] 3.5 Detection of inflammatory factors Cell supernatant was collected, and the TNF-α inflammatory factor level was detected according to the ELISA kit instructions. The TNF-α inhibition rate was then calculated using the formula: TNF−α inhibition rate (%) = (1 − TNF−α concentration in the sample group / TNF−α concentration in the LPS group) × 100%. The results are shown in Table 5.

[0055] 4. In vitro antioxidant test (DPPH test) 4.1 Experimental Principle A higher DPPH free radical scavenging rate indicates a better antioxidant effect.

[0056] 4.2 Test Methods 4.2.1 Preparation of DPPH solution Weigh 4 mg of DPPH powder, dissolve it in 60% v / v ethanol aqueous solution, and make up to 100 mL to prepare a DPPH solution with a concentration of 40 mg / L. Store in the dark. 4.2.2 Sample solution preparation Using compositions 1-11 as samples, weigh the corresponding samples, dissolve and dilute them with 60% v / v ethanol aqueous solution to prepare a sample solution with a concentration of 5 mg / mL; 4.2.3 Experimental Grouping Blank control group (A0): 2 mL sample solution + 2 mL 60% v / v ethanol aqueous solution; Sample group (A1): 2 mL sample solution + 2 mL DPPH solution; Sample matrix group (A2): 2 mL DPPH solution + 2 mL 60% v / v ethanol aqueous solution; Reaction: After mixing each group evenly, let it stand in the dark for 30 minutes, centrifuge at 5000 r / min for 10 minutes, take the supernatant, and measure the absorbance at a wavelength of 517 nm.

[0057] 4.2.4 Parallel Experiments and Replication Three parallel tubes were set up for each sample, and the average value of the results was taken.

[0058] 4.3 Data Calculation The formula for DPPH free radical scavenging rate is: Scavenging rate (%) = [1−(A1−A0) / A2]×100%, and the results are shown in Table 5.

[0059] 5. Results Table 5 Results of each group of samples Group / Project DPPH free radical scavenging rate / % IL-6 inhibition rate / % TNF-α inhibition rate / % Composition 1 58.72 35.69 38.71 Composition 2 72.65 42.18 45.33 Composition 3 89.27 56.39 60.29 Composition 4 86.41 51.72 57.84 Composition 5 81.39 47.65 53.17 Composition 6 75.82 43.29 49.62 Composition 7 42.35 22.17 31.42 Composition 8 45.68 24.35 33.58 Composition 9 38.52 21.34 18.76 Composition 10 55.19 33.42 36.25 Composition 11 78.53 45.26 51.37 6. Results Analysis According to the test results, compositions 1-6 have excellent performance in anti-oxidation, anti-aging, and anti-inflammatory effects. Compositions 7-10 are single-component missing control formulations of composition 3. The results show that compositions 7-10 have significantly lower antioxidant, anti-aging, and anti-inflammatory effects compared to composition 3. This result proves that fisetin, acetyl hexapeptide-1, tuberose polysaccharide, and European chestnut leaf extract have irreplaceable complementary and synergistic effects, solving the problem that traditional anti-aging formulas cannot balance irritation and efficacy.

[0060] Composition 11 replaces fisetin ①, prepared by dynamic countercurrent extraction combined with macroporous resin column purification, with fisetin ②, prepared by traditional organic solvent thermal reflux extraction. Test results show that the purity of fisetin ① is higher than that of fisetin ②, and the three efficacy indicators of composition 11 are all lower than those of composition 3. This proves that the preparation process of dynamic countercurrent extraction combined with macroporous resin column purification used in this invention can extract and retain the bioactivity of fisetin more efficiently, avoid the degradation of active ingredients caused by high temperature in the traditional thermal reflux extraction process, and at the same time, this process is green and environmentally friendly, with low organic solvent residue, which better meets the safety requirements of cosmetic raw materials.

[0061] V. A lotion that removes dead skin cells and improves the appearance of aging skin in sensitive skin. The specific preparation steps of the emulsion are as follows: Step 1: Mix the thickener and humectant evenly, add pure water, heat at 75℃ water bath temperature for 10 minutes, then homogenize at 4000rpm for 5 minutes, keep warm and set aside to obtain pre-prepared component A.

[0062] Step 2: Take the oil and emulsifier in a beaker, heat to 75°C, dissolve evenly, keep warm for later use, and obtain the pre-prepared component B.

[0063] Step 3: Heat the pre-made component A to 75°C, add component B at 75°C in a homogenizer at 4000 rpm, homogenize for 5 min, and obtain the emulsified base material.

[0064] Step 4: Stir and cool the emulsified base material to 50°C, add composition 3, continue stirring until the material is uniform, then add preservative and pH adjuster to adjust the pH to 6.0-6.5, stop stirring, discharge the material, and obtain an emulsion that removes aging skin cells and improves the skin aging of sensitive skin.

[0065] The amounts of each ingredient added to the above emulsion are detailed in Table 6.

[0066] Table 6 Raw material addition amount (unit: wt%)

[0067] VI. Human Efficacy Testing of Emulsion 1. Human patch test 1.1 Test Method Thirty participants were selected for the test according to the inclusion criteria, and the sample size was no more than 50 mm². 2A qualified spot test apparatus with a depth of approximately 1 mm is used. The sample is placed in the small chamber of the spot test apparatus, with a volume of approximately 0.020 mL to 0.025 mL. The spot test apparatus containing the sample (Application Examples 1, 2, and 3: matrix group; blank control group: distilled water) is applied to the flexor side of the subject's forearm using hypoallergenic adhesive tape. Gently press with the palm of the hand to ensure even application to the skin, and leave for 24 hours. Skin reactions are observed according to the standards in Table 7 at 30 minutes (after the indentation disappears), 24 hours, and 48 hours after removing the test apparatus, and the results are recorded.

[0068] Table 7 Adverse Reaction Rating Rating levels Skin reaction 0 negative reaction 1 Suspicious reaction, only slight erythema 2 Weak positive reaction (erythema reaction): erythema, infiltration, edema, and papules may be present. 3 Strong positive reaction (herpes reaction): erythema, infiltration, edema, papules; the reaction may extend beyond the test area. 4 Extremely strong positive reaction (confluent herpes simplex reaction): obvious erythema, severe infiltration, edema, confluent herpes simplex; reaction extends beyond the test area. 1.2 Result Judgment Interpretation of results from occlusive skin patch testing: If more than 5 out of 30 subjects have a suspected Grade 1 skin reaction, or more than 2 out of 30 subjects have a weakly positive Grade 2 skin reaction, or if any one subject has a Grade 3 or higher adverse skin reaction, the test substance is considered to have an adverse skin reaction on humans.

[0069] After testing, the application examples 1-3 and the matrix emulsion of the present invention all showed negative reactions after human patch testing, indicating that they are safe and non-irritating to human skin.

[0070] 2. Human efficacy test 2.1 Test Method Forty participants (healthy women aged 30-40) were selected according to the subject inclusion criteria and randomly divided into four groups of 10 people each. Each group was assigned a trial sample (emulsions from Application Examples 1-3 and the matrix group). Sample usage method: After cleansing the face, the subject should take 1g of sample lotion, apply it evenly to the face, and gently massage for 2 minutes to allow the skin to absorb it. Use it once in the morning and once in the evening for a total of 28 days.

[0071] The Tewameter®™ HEX epidermal water loss meter and Dermatop instrument were used to test the facial skin condition before product use (day 0), and on days 14 and 28 after product use, so as to comprehensively evaluate the repair, firming and anti-wrinkle effects of the product.

[0072] The specific experimental parameters and equipment used in the experiment are shown in Table 8.

[0073] Table 8 Test Parameters and Equipment

[0074] The formulas for calculating changes in TWEL and the number of wrinkles are as follows: TWEL improvement rate = (T0 sample group - T14, 28 sample groups) / T0 sample group * 100%; Wrinkle improvement rate = (T0 sample group - T14 and 28 sample groups) / T0 sample group * 100%; Firmness improvement rate = (T14, 28 sample groups - T0 sample group) / T0 sample group * 100%.

[0075] 2.2 Test Results The experimental results are shown in Table 9; Table 9 Results of Human Efficacy Tests

[0076] 2.3 Results Analysis According to the test results, the lotion containing the anti-aging repair composition provided by the present invention is safe, non-irritating and non-allergenic to human skin, and is suitable for the long-term use needs of people with sensitive skin. According to the test results in Table 9, the composition of the present invention can significantly improve the barrier function of sensitive skin by inhibiting skin inflammation and repairing damaged stratum corneum, reducing transepidermal water loss, and fundamentally relieving discomfort symptoms such as redness, dryness, itching, and stinging of sensitive skin. It has excellent skin repair effects. At the same time, the composition of the present invention can effectively improve skin firmness, reduce the number of facial wrinkles, and delay the skin aging process by inhibiting the aging of dermal fibroblasts and reducing the secretion of aging-related pro-inflammatory factors. It has a safe and effective anti-aging effect for people with sensitive skin.

[0077] While specific embodiments of the present invention have been described above, those skilled in the art should understand that these are merely illustrative examples, and the scope of protection of the present invention is defined by the appended claims. Those skilled in the art can make various changes or modifications to these embodiments without departing from the principles and essence of the present invention, but all such changes and modifications fall within the scope of protection of the present invention.

Claims

1. An anti-aging repair composition, characterized in that, The composition comprises, by weight parts, the following components: 0.01-1 parts of fisetin, 0.01-1.5 parts of acetyl hexapeptide-1, 0.01-0.3 parts of tuberose polysaccharide, and 1-4 parts of European chestnut leaf extract.

2. The composition according to claim 1, characterized in that, The composition comprises, by weight parts, the following components: 0.1-0.8 parts of fisetin, 0.1-0.8 parts of acetyl hexapeptide-1, 0.1-0.2 parts of tuberose polysaccharide, and 1.1-3 parts of European chestnut leaf extract.

3. The composition according to claim 2, characterized in that, The composition comprises, by weight parts, the following components: 0.4-0.6 parts of fisetin, 0.4-0.6 parts of acetyl hexapeptide-1, 0.14-0.16 parts of tuberose polysaccharide, and 1.9-2.1 parts of European chestnut leaf extract.

4. The composition according to any one of claims 1-3, characterized in that, The method for preparing the fisetin includes the following steps: Step 1: Crush and sieve the dried sawdust from the wax tree to obtain wax tree powder for later use; Step 2: Dynamic countercurrent extraction of wax tree powder to obtain an extract containing the target compound; Step 3: Concentrate the extract containing the target compound to obtain a crude extract of fisetin. Step 4: Purify and separate the crude extract of fisetin using a macroporous adsorption resin column, and collect the target eluent; Step 5: Concentrate the target eluent under reduced pressure and recrystallize at a lower temperature to obtain fisetin crystals; Step 6: Vacuum dry the fisetin crystals to obtain fisetin.

5. The composition according to claim 4, characterized in that, In step 2, dynamic countercurrent extraction specifically involves using 60%-70% ethanol aqueous solution as solvent and performing 3-5 stages of continuous countercurrent extraction at 60-75℃ and a material-to-liquid ratio of 1:8-1:

12.

6. The composition according to claim 4, characterized in that, In step 3, the concentration specifically involves concentrating the extract under reduced pressure at a vacuum of -0.08 to -0.09 MPa and a temperature of 55-65°C, recovering the ethanol, and obtaining a crude extract.

7. The composition according to claim 4, characterized in that, In step 4, the purification and separation specifically involves redissolving the crude extract in water at a mass-to-volume ratio of 1:3 to 1:5 g / mL, filtering to obtain a clear sample solution, and loading the clear solution onto a macroporous adsorption resin column at a flow rate of 1 to 2 BV / h. The sample volume is 0.3 to 0.5 BV of the resin's wet volume. After loading, the column is eluted with 3 to 5 BV of pure water at a flow rate of 2 to 3 BV / h to remove water-soluble impurities. After elution, the column is eluted again with 3 to 5 BV of 70 v / v% to 80 v / v% ethanol aqueous solution at a flow rate of 1 to 2 BV / h, and the eluent rich in fisetin is collected.

8. The composition according to claim 4, characterized in that, In step 5, the cooling recrystallization specifically involves concentrating the eluent under reduced pressure at 50-60°C to 1 / 5-1 / 10 of its original volume, then cooling it to 0-4°C via a programmed cooling process for recrystallization, followed by filtration to obtain crystals.

9. The use of the composition according to any one of claims 1-7 in the preparation of skin care products with anti-inflammatory, antioxidant and anti-aging effects.

10. A lotion that removes aging skin cells and improves skin aging in sensitive skin, characterized in that, The emulsion contains the composition according to any one of claims 1-7.

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