Tightening and anti-wrinkle composition as well as preparation method and application thereof

Abstract

The invention discloses a firming and anti-wrinkle composition as well as a preparation method and application thereof, and the firming and anti-wrinkle composition comprises the following components in parts by mass: 0.2-0.5 part of a growth factor accelerant, 1 part of a lindera glauca fruit extract and 0.02-0.08 part of salmon essence source DNA (deoxyribonucleic acid) sodium. The growth factor accelerant is selected from at least one of hydroxyl pinacolone retinoate, bakuchiol, a spanishneedles herb extract, asiaticoside, palmitoyl tripeptide-1 and palmitoyl tetrapeptide-7. The growth factor accelerant, the lindera glauca fruit extract and the salmon essence source DNA sodium cooperate to improve the problems of skin laxity and slowdown epidermal layer cell renewal caused by deep protein loss and wrinkle formation caused by decline of the stratum corneum moisturizing capacity, and the effects of tightening and resisting wrinkles are synergistically exerted in multiple aspects.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, specifically relating to a firming and anti-wrinkle composition, its preparation method, and its application. Background Technology

[0002] As skin ages, it undergoes aging, manifesting as sagging and wrinkles. This is due to the collapse of the skin's deep supporting structures (decreased firmness) and the formation of epidermal folds (wrinkles), though these two causes are not entirely related. Currently, most research and commercially available products target stimulating collagen synthesis to achieve firming and anti-wrinkle effects. However, this approach fails to address the underlying skin aging caused by the collapse of the deep supporting structures and neglects the crucial role of cell renewal and inhibiting protein breakdown in anti-wrinkle efforts. Elastin and fibrillin, like "springs" in a "bed mattress" (dermal matrix), support the skin's elasticity and resilience, achieving deep firmness. Therefore, upregulating elastin synthesis is essential for deep skin firming. Conversely, inhibiting the breakdown and loss of elastin and fibrillin can boost elastin levels. Furthermore, promoting epidermal cell renewal and hydration are equally important for smoothing wrinkles and inhibiting fold formation.

[0003] Therefore, this invention is proposed. Summary of the Invention

[0004] This invention provides a firming and anti-wrinkle composition, its preparation method, and its application, aiming to simultaneously improve skin laxity caused by deep protein loss and wrinkle formation caused by slowed epidermal cell renewal and decreased stratum corneum moisturizing ability.

[0005] This invention provides the following technical solution: In a first aspect, the present invention provides a firming and anti-wrinkle composition, comprising, by weight parts: 0.2-0.5 parts of a growth factor promoter, 1 part of *Litsea cubeba* (… Lindera glauca The product contains fruit extract and 0.02-0.08 parts of salmon extract-derived DNA sodium; the growth factor promoter is selected from at least one of hydroxypinazone retinate, psoralen, Bidens trifoliata extract, asiaticoside, palmitoyl tripeptide-1 and palmitoyl tetrapeptide-7.

[0006] Growth factor promoters include those that can promote the expression and secretion of TGF-β (such as hydroxypinazone retinate, psoralen, Bidens pilosa extract, asiaticoside, etc.) or mimic the function of TGF-β (such as palmitoyl tripeptide-1, palmitoyl tetrapeptide-7), thereby acting on the TGF-β / Smad signaling pathway and promoting elastin synthesis.

[0007] Mountain pepper fruit (Latin name) Linderaglauca(Siebold&Zucc.)BlumeThe extract is rich in volatile oils (such as citral, β-cis-ocimene, caryophyllene, β-pinene, D-gemmaene, etc.), alkaloids (such as isoquinoline alkaloids), organic acids, flavonoids, and other flavonoid substances. In the dermis, it can gently inhibit the excessive activity of matrix metalloproteinases (MMP-1, MMP-3, MMP-9), reduce the degradation of collagen and elastin, and protect the existing support structure. In the epidermis, it promotes epidermal cell renewal and differentiation by activating specific receptors on keratinocytes (such as subtypes of TRPV ion channels), increasing epidermal thickness and thus smoothing fine wrinkles. Therefore, the extract of *Litsea cubeba* fruit can achieve a firming and anti-wrinkle effect by protecting the existing structure, reducing elastin degradation, and reducing the formation of epidermal folds (wrinkles). Combined with growth factor promoters (which can upregulate elastin synthesis), it synergistically achieves a firming and anti-wrinkle effect from three levels (upregulating elastin synthesis, reducing elastin degradation, and promoting epidermal cell renewal and differentiation).

[0008] Salmon DNA sodium is a sodium salt of DNA fragments (deoxyribonucleic acid) extracted from the testes (white part) of salmon. It has the effects of deep repair and regeneration, moisturizing, anti-sun aging and strengthening the skin barrier. It has a strong cell proliferation promotion effect and moisturizing effect. Together with growth factor promoters and pepper fruit extract, it can tighten the skin and smooth wrinkles.

[0009] In some embodiments, the method for extracting the Litsea cubeba fruit extract includes the following steps: The surfactant and water are mixed to obtain an entrainer, which is then added to a solvent tank. The mountain pepper fruits were washed, dried, and crushed to obtain mountain pepper fruit powder, which was then placed in an extraction vessel. Supercritical CO2 fluid is introduced through a solvent vessel and then into an extraction vessel for extraction reaction. Cooling and depressurization recover CO2, yielding an oil-water mixture; Preheated water was added to the oil-water mixture, and homogenization and phase inversion were performed to obtain an emulsion of Litsea cubeba fruit extract.

[0010] The method for preparing Litsea cubeba fruit extract provided by this invention can simultaneously increase the content of fat-soluble volatile oils, flavonoids, and water-soluble alkaloids and polyphenols. In contrast, existing technologies using alcohol or water extraction cannot avoid drying and concentrating the crude extract, leading to the loss of volatile oils. The monoterpenes and sesquiterpenes in the volatile oils possess broad-spectrum antibacterial, anti-allergic, and skin-smoothing effects; therefore, increasing the content of volatile oils in the extract is beneficial for enhancing its firming and anti-wrinkle effects.

[0011] This invention employs surfactant-assisted supercritical CO2 extraction to obtain *Litsea cubeba* fruit extract. The extraction process utilizes the formation of a microemulsion between the supercritical fluid, surfactant, and water to improve the extraction rate of water-soluble components, while the more lipophilic supercritical CO2 fluid ensures the extraction of fat-soluble components. Simultaneously, the surfactant used can serve as an emulsifier for the subsequent preparation of the extract emulsion. A stable oil-in-water emulsion is prepared via a phase-inversion emulsion method, avoiding the need for drying and concentration steps. Furthermore, this emulsion exhibits good room-temperature storage stability, allowing the fat-soluble active ingredients to be stably dispersed in the aqueous phase, maintaining their activity, and preventing stratification that could negatively impact the user experience.

[0012] In some embodiments, the surfactant is at least one selected from PEG-30 dimerhydroxystearate, sorbitan monopalmitate, sorbitan monolaurate, and polyglycerol-2-didihydroxystearate. The HLB value and PIT temperature (phase transition temperature) of the surfactant affect the stability of the resulting emulsion and the preparation process conditions. This invention selects surfactants with an HLB value of 5-8.6, which are near the critical point between hydrophilic and lipophilic interactions, thus exhibiting good emulsifying effects in both extraction systems and the preparation of oil-in-water emulsions. These surfactants have a PIT temperature of 35-60°C, a relatively low phase transition temperature, thus eliminating the need for high temperatures to complete the transformation from water-in-oil to oil-in-water, avoiding the loss of volatile oils.

[0013] In some embodiments, the mass ratio of the surfactant to water is (10:90) to (3:97).

[0014] In some implementations, the crushing refers to crushing the pepper fruit to 60-80 mesh.

[0015] In some embodiments, the ratio of the pepper fruit to the entrainer is 1 kg: 100-300 mL. If the amount of entrainer is too small, it is difficult to improve the extraction rate of water-soluble active ingredients; if the amount of entrainer is too large, it is easy to reduce the compatibility and non-polarity of the extraction fluid, resulting in a decrease in the extraction rate.

[0016] In some implementations, the extraction pressure is 20-30 MPa, the temperature is 40-60 °C, and the supercritical CO2 flow rate is 20-30 kg / h.

[0017] In some embodiments, the mixing ratio of the pepper fruit powder and supercritical CO2 is 1g:2~4mL.

[0018] In some implementations, the cooling and decompression refers to reducing the pressure to 5-10 MPa and / or reducing the temperature to 25°C-40°C.

[0019] In some embodiments, the preheated water temperature is 40~60°C; selecting an appropriate temperature for homogenization and phase inversion based on the PIT temperature of the surfactant is beneficial to improving the stability of the extract emulsion.

[0020] In some embodiments, the amount of preheated water added is determined according to the water content of the pepper fruit extract being 40% to 60%.

[0021] In some implementations, the homogenization phase inversion refers to homogenization at 800-1200 rpm.

[0022] In some embodiments, the firming and anti-wrinkle composition further includes one or more of the following: soluble proteoglycans, serum albumin, fibronectin, collagen, chitosan, Inonotus obliquus extract, Betula alba sap, Bletilla striata root water, ceramide, and sodium polyaspartate.

[0023] In some embodiments, the firming and anti-wrinkle composition comprises, by weight percentage: 0.2%–3% hydroxypinazone retinate, 0.2%–4% asiaticoside, 1%–5% palmitoyl tripeptide-1, 20%–30% peperomia fruit extract and 0.5%–1% salmon extract-derived DNA sodium, with the balance being betula alba sap.

[0024] In some embodiments, the firming and anti-wrinkle composition comprises, by weight percentage: 0.2%–3% hydroxypinazone retinate, 1%–2% psoralen, 5%–10% Bidens trifoliata extract, 20%–30% Litsea cubeba fruit extract, 0.5%–1% salmon extract-derived sodium DNA, 2%–3% soluble proteoglycans, 0.5%–3% chitosan, and 5%–10% Inonotus obliquus extract, with the balance being water.

[0025] In some embodiments, the firming and anti-wrinkle composition is applied to a serum, face cream, lotion, or mask.

[0026] Secondly, the present invention provides a firming and anti-wrinkle serum, wherein the serum comprises, by weight percentage, the following components: 10% to 30% of any of the firming and anti-wrinkle compositions described above, 7% to 15% of a moisturizer, 0.3% to 1.5% of a preservative, 0% to 7% of a thickener, and 0.01% to 0.05% of a pH adjuster, with the balance being water.

[0027] In some embodiments, the moisturizer is selected from at least one of glycerin, butylene glycol, dipropylene glycol, propylene glycol, pentanediol, and sodium hyaluronate.

[0028] In some embodiments, the preservative is selected from at least one of p-hydroxyacetophenone, phenoxyethanol, and ethylhexylglycerin.

[0029] In some embodiments, the thickener is selected from at least one of carbomer and xanthan gum.

[0030] In some embodiments, the pH adjuster is selected from at least one of citric acid, lactic acid, and triethanolamine.

[0031] Thirdly, the present invention also provides a method for preparing the above-mentioned firming and anti-wrinkle serum, characterized by comprising the following steps: (1) Dissolve the thickener in water, heat to 50~60℃ to dissolve, keep warm for later use, and obtain solution A; (2) Add humectant and preservative to solution A, stir evenly, and then cool to 20~30℃ to obtain solution B; (3) Add the firming and anti-wrinkle composition to the solution B, stir evenly, add pH adjuster and defoamer to obtain firming and anti-wrinkle essence.

[0032] Beneficial effects:

[0033] This invention provides a firming and anti-wrinkle composition, its preparation method, and its application. The composition contains a growth factor promoter, a Sichuan pepper fruit extract, and salmon extract-derived sodium DNA. These three components synergistically improve skin laxity caused by deep protein loss and wrinkle formation due to slowed epidermal cell renewal and decreased stratum corneum moisturizing ability, thus exerting a multi-faceted firming and anti-wrinkle effect. The Sichuan pepper fruit extract provided by this invention is obtained through surfactant-assisted supercritical CO2 extraction, which can simultaneously increase the content of fat-soluble volatile oils, flavonoids, and water-soluble alkaloids and polyphenols. Attached Figure Description

[0034] Figure 1 To produce physical photographs of the subject's eye before and after the experiment. Detailed Implementation

[0035] The present invention will be further described in detail below with reference to specific embodiments, but the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in this technical field.

[0036] Example 1 This embodiment provides a firming and anti-wrinkle composition comprising, by weight percentage: 1.4% hydroxypinazone retinate, 2% asiaticoside, 3% palmitoyl tripeptide-1, 20% peperomia fruit extract and 0.8% salmon extract-derived DNA sodium, with the balance being betula alba sap.

[0037] The preparation method of Litsea cubeba fruit extract includes the following steps: (1) The entrainer was obtained by mixing dehydrated sorbitan monopalmitate and water at a mass ratio of 6:94 and then adding it to the solvent tank; (2) Wash the fresh mountain pepper fruit, then dry it under hot air at 40℃ for three days. After crushing, pass it through an 80-mesh sieve to obtain mountain pepper fruit powder, which is placed in an extraction vessel. The ratio of mountain pepper fruit powder to entrainer is 1kg:200mL. (3) Supercritical CO2 fluid is introduced into the solvent vessel and then into the extraction vessel for extraction; the pressure during extraction is 25 MPa, the temperature is 50℃, the flow rate of supercritical CO2 is 25 kg / h, and the mixing ratio of pepper fruit powder and supercritical CO2 is 1 g: 3 mL. (4) Reduce the pressure to 5 MPa to recover CO2, collect the oil-water mixture in the separation vessel, test the water content in the oil-water mixture (weight loss rate after drying at 80℃ for 24h), and then add ultrapure water preheated to 50℃ at a rate of 1L / min according to the water content of 45% in the final product of the mountain pepper fruit extract. Homogenize at 1000 rpm until the temperature drops to 30℃ to obtain the emulsion-like mountain pepper fruit extract.

[0038] The method for preparing the above-mentioned firming and anti-wrinkle composition includes the following steps: (1) Dissolve asiaticoside in the sap of birch (BETULA ALBA) at 50℃ to obtain phase A, and keep it warm for later use; (2) Add the pepper fruit extract to phase A while stirring. After the addition is complete, lower the temperature to 25°C. Finally, add hydroxypinazone retinate and salmon extract-derived DNA sodium, stir evenly, and degas to obtain a firming and anti-wrinkle composition.

[0039] Partial source information: Palmitoyl tripeptide-1, purchased from Jining Fangyu Chemical Co., Ltd.; fresh Sichuan pepper fruit, purchased from Dazhou Agricultural Market, Sichuan; sodium DNA, a salmon extract source, purchased from Xi'an Minglang Biotechnology Co., Ltd.; medical grade birch sap, purchased from Shaanxi Yangyuanshen Biotechnology Co., Ltd.

[0040] Example 2

[0041] The difference from Example 1 is that the preparation method of the Litsea cubeba fruit extract is as follows: (1) Wash the fresh mountain pepper fruit, then dry it under hot air at 40℃ for three days, crush it and pass it through an 80-mesh sieve to obtain mountain pepper fruit powder; (2) Soak the pepper fruit powder in ethanol at 40℃ for 36 hours and repeat 3 times. Combine the extracts and distill under reduced pressure at 60℃ to obtain the extract. The mixing ratio of pepper fruit powder and ethanol is 1kg:2L. (3) Add 3% of dehydrated sorbitan monopalmitate by weight of the extract, and then add ultrapure water preheated to 50°C at a rate of 1L / min according to the water content of the final product of the pepper fruit extract being 45%. Homogenize at 1000rpm until the temperature drops to 30°C to obtain the pepper fruit extract.

[0042] Example 3

[0043] The difference from Example 1 is that the preparation method of the Litsea cubeba fruit extract is as follows: (1) Add ethanol as an entrainer to the solvent tank; (2) Wash the fresh mountain pepper fruit, then dry it under hot air at 40℃ for three days. After crushing, pass it through an 80-mesh sieve to obtain mountain pepper fruit powder, which is placed in an extraction vessel. The ratio of mountain pepper fruit powder to entrainer is 1kg:200mL. (3) Supercritical CO2 fluid is introduced into the solvent vessel and then into the extraction vessel for extraction; the pressure during extraction is 25 MPa, the temperature is 50℃, the flow rate of supercritical CO2 is 25 kg / h, and the mixing ratio of pepper fruit powder and supercritical CO2 is 1 g: 3 mL. (4) Reduce the pressure to 5 MPa to recover CO2 and obtain crude product of Litsea cubeba fruit extract; (5) The crude product of the Litsea cubeba fruit extract was distilled under reduced pressure at 50°C for 2 hours to obtain oil. After cooling, 3% of the oil mass of dehydrated sorbitan monopalmitate was added. Then, ultrapure water preheated to 50°C was added at a rate of 1L / min, according to the water content of the final product of the Litsea cubeba fruit extract being 45%. The mixture was homogenized at 1000rpm until the temperature dropped to 30°C to obtain the Litsea cubeba fruit extract.

[0044] Example 4

[0045] The difference from Example 1 is that in the preparation method of the Litsea cubeba fruit extract, an equal amount of AOT (sodium di(2-ethylhexyl) succinate sulfonate) is used as the surfactant instead of dehydrated sorbitan monopalmitate.

[0046] Example 5

[0047] The difference from Example 1 is that the preparation method of the Litsea cubeba fruit extract is as follows: The preparation method of Litsea cubeba fruit extract includes the following steps: (1) Mix AOT and water at a mass ratio of 6:94 to obtain an entrainer; (2) Wash the fresh mountain pepper fruit, then dry it under hot air at 40℃ for three days. After crushing, pass it through an 80-mesh sieve to obtain mountain pepper fruit powder. Soak the powder in the entrainer for 12 hours, and then put it into the extraction vessel. The ratio of mountain pepper fruit powder to entrainer is 1kg:600mL. (3) Supercritical CO2 fluid is introduced into the solvent vessel and then into the extraction vessel for extraction; the pressure during extraction is 25 MPa, the temperature is 50℃, the flow rate of supercritical CO2 is 25 kg / h, and the mixing ratio of pepper fruit powder and supercritical CO2 is 1 g: 3 mL. (4) Reduce the pressure to 5 MPa to recover CO2 and collect the crude product of Litsea cubeba fruit extract in the separation vessel; (5) The crude product of Litsea cubeba fruit extract was distilled under reduced pressure at 80°C for 2 hours. After cooling, ultrapure water preheated to 50°C was added at a rate of 1L / min according to the water content of the final product of Litsea cubeba fruit extract being 45%. The mixture was homogenized at 1000rpm until the temperature dropped to 30°C to obtain Litsea cubeba fruit extract.

[0048] Example 6 This embodiment provides a firming and anti-wrinkle composition comprising, by weight percentage: 0.2% hydroxypinazone retinate, 4% asiaticoside, 1% palmitoyl tripeptide-1, 25% peperomia fruit extract and 2% salmon extract-derived DNA sodium, with the balance being birch (BETULA ALBA) sap.

[0049] The preparation method of Litsea cubeba fruit extract includes the following steps: (1) The entrainer was obtained by mixing dehydrated sorbitan monolaurate, polyglycerol-2-dimeric hydroxystearate and water in a mass ratio of 3:2:95 and then adding it to the solvent tank; (2) Wash the fresh mountain pepper fruit, then dry it under hot air at 40℃ for three days. After crushing, pass it through an 80-mesh sieve to obtain mountain pepper fruit powder, which is then placed in an extraction vessel. The ratio of mountain pepper fruit powder to entrainer is 1kg:100mL. (3) Supercritical CO2 fluid is introduced into the solvent vessel and then into the extraction vessel for extraction; the pressure during extraction is 20 MPa, the temperature is 50 ℃, the flow rate of supercritical CO2 is 30 kg / h, and the mixing ratio of pepper fruit powder and supercritical CO2 is 1 g: 2 mL. (4) Reduce the pressure to 5 MPa to recover CO2, collect the oil-water mixture in the separation vessel, test the water content in the oil-water mixture (weight loss rate after drying at 80℃ for 24h), and then add ultrapure water preheated to 50℃ at a rate of 1L / min according to the water content of the final product of the mountain pepper fruit extract being 55%, and maintain homogenization at 1000rpm until the temperature drops to 30℃, thus obtaining the emulsion-like mountain pepper fruit extract.

[0050] The preparation method of the above-mentioned firming and anti-wrinkle composition is the same as that in Example 1.

[0051] Example 7 This embodiment provides a firming and anti-wrinkle composition comprising, by weight percentage: 3% hydroxypinazone retinate, 3% psoralen, 8% Bidens trifoliata extract, 30% Ligusticum chuanxiong fruit extract, 0.6% salmon extract-derived DNA sodium, 2.5% soluble proteoglycan, 1.7% chitosan, and 6% Inonotus obliquus extract, with the balance being water.

[0052] The preparation method of Litsea cubeba fruit extract includes the following steps: (1) Mix PEG-30 dihydroxystearate and water at a mass ratio of 3:97 to obtain an entrainer, and then add it to a solvent tank; (2) Wash the fresh mountain pepper fruit, then dry it under hot air at 40℃ for three days. After crushing, pass it through an 80-mesh sieve to obtain mountain pepper fruit powder, which is then placed in an extraction vessel. The ratio of mountain pepper fruit powder to entrainer is 1kg:300mL. (3) Supercritical CO2 fluid is introduced into the solvent vessel and then into the extraction vessel for extraction; the pressure during extraction is 25 MPa, the temperature is 50℃, the flow rate of supercritical CO2 is 25 kg / h, and the mixing ratio of pepper fruit powder and supercritical CO2 is 1 g: 3 mL. (4) Reduce the pressure to 5 MPa to recover CO2, collect the oil-water mixture in the separation vessel, test the water content in the oil-water mixture (weight loss rate after drying at 80℃ for 24h), and then add ultrapure water preheated to 60℃ at a rate of 1L / min according to the water content of 45% in the final product of the mountain pepper fruit extract, and maintain homogenization at 1000 rpm until the temperature drops to 30℃, thus obtaining the emulsion-like mountain pepper fruit extract.

[0053] The method for preparing the above-mentioned firming and anti-wrinkle composition includes the following steps: (1) Psoralen, Bidens trifoliata extract, soluble proteoglycan, Inonotus obliquus extract and chitosan were dissolved in ultrapure water at 50°C to obtain phase A, which was kept warm for later use. (2) Add the pepper fruit extract to phase A while stirring. After the addition is complete, lower the temperature to 25°C. Finally, add hydroxypinazone retinate and salmon extract-derived DNA sodium, stir evenly, and degas to obtain a firming and anti-wrinkle composition.

[0054] Partial source information for raw materials: Bidens pilosa extract, water concentrate, purchased from Lanzhou Waterles Biotechnology Co., Ltd.; soluble proteoglycans, purchased from Hubei Langbowan Biomedical Co., Ltd.; chitosan, medical grade, purchased from Qingdao Yuekang Biotechnology Co., Ltd.; Inonotus obliquus extract, water concentrate, purchased from Baoji Liupanyun Biotechnology Co., Ltd.

[0055] Comparative Example 1 The difference from Example 1 is that no growth factor promoter is added.

[0056] A composition comprising, by weight percentage: 20% Litsea cubeba fruit extract, 0.8% sodium DNA derived from salmon extract, and the balance being BETULA ALBA sap.

[0057] Comparative Example 2 The difference from Example 1 is that no pepper fruit extract was added.

[0058] A composition comprising, by weight percentage: 1.4% hydroxypinazone retinate, 2% asiaticoside, 3% palmitoyl tripeptide-1, 0.8% salmon extract-derived DNA sodium, with the remainder being birch (BETULA ALBA) sap.

[0059] Comparative Example 3 The difference from Example 1 is that no pepper fruit extract was added.

[0060] A composition comprising, by weight percentage: 1.4% hydroxypinazone retinyl ester, 2% asiaticoside, 3% palmitoyl tripeptide-1 and 20% Litsea cubeba fruit extract, with the balance being betula alba sap.

[0061] Test Example 1: Emulsion Stability To investigate the stability of the emulsion-like extract of Litsea cubeba fruit, the following tests were conducted: Sealed samples were placed in an environment of -10±1℃ for two weeks, and after returning to room temperature, their appearance was compared with samples stored at room temperature (25±3℃). The average particle size of the emulsion was measured using a dynamic light scattering instrument to evaluate cold resistance. Sealed samples were placed in an environment of 48±1℃ for two weeks, and after returning to room temperature, their appearance was compared with samples stored at room temperature (25±3℃). The average particle size of the emulsion was measured using a dynamic light scattering instrument to evaluate heat resistance. The results are shown in Table 1.

[0062] Table 1

[0063] As shown in Table 1, the Litsea cubeba fruit extract prepared by the preparation method provided by the present invention is in the form of an emulsion and has good stability.

[0064] Test Example 2 Antioxidant Test The extract of *Litsea cubeba* fruit was dissolved in ultrapure water to obtain an extract solution with a solid content (obtained by drying at 80℃ for 24 h) of 100 mg / L. A DPPH free radical scavenging ability assay kit (purchased from Shanghai Enzyme-Link Biotechnology Co., Ltd.) was used, and the experiment was conducted according to the kit instructions. A control group, experimental group, positive group, and blank group were set up, and the test solutions were prepared according to the groupings shown in Table 2 below. The absorbance value of each tube was measured at a wavelength of 517 nm. A 100 mg / L vitamin C solution was used as a positive control. The DPPH free radical scavenging rate of each sample was calculated according to the following formula: DPPH free radical scavenging rate (%) = [1 - (A1 - A2) ÷ A...] 空白组 ]×100%, where A1 is the absorbance value of the experimental group or positive group, A2 is the absorbance value of the experimental group control or positive group control, A 空白组 The absorbance values ​​are for the blank tubes. The average DPPH radical scavenging rates of the *Litsea cubeba* fruit extracts from each example are shown in Table 3.

[0065] Table 2

[0066] Table 3

[0067] Two-tailed tests were used for statistical analysis, and independent samples t-tests were used to calculate p-values. The p-values ​​were compared with the model control. *: p < 0.05 indicates a significant difference, and **: p < 0.01 indicates a highly significant difference.

[0068] As shown in Table 3, the extract of Litsea cubeba has good antioxidant activity, which mainly comes from flavonoids and phenolic acids. Furthermore, the antioxidant activity of the extract obtained by the surfactant-assisted supercritical CO2 extraction method provided by this invention is similar to that obtained by the alcohol extraction method.

[0069] Test Example 3: Volatile Oil Content Test The volatile oil content of the extracts of Litsea cubeba fruit obtained by different extraction methods was determined according to Method A of the volatile oil determination method in the 2020 edition of the Pharmacopoeia of the People's Republic of China. The total amount of volatile oil in the extracts obtained by different extraction methods was determined using 1 kg of Litsea cubeba fruit powder extracted according to the methods of different embodiments. The results are shown in Table 4.

[0070] Table 4

[0071] As shown in Table 4, surfactant-assisted supercritical CO2 extraction improves the extraction rate of volatile oils compared to alcohol extraction (Example 2) and ethanol-assisted supercritical CO2 extraction, thereby enhancing the medicinal activity of the extract. Examples 4 and 5 used more hydrophilic surfactants, but their volatile oil yields were lower because the affinity of more hydrophilic surfactants for supercritical CO2 fluid is weakened.

[0072] Test Example 4: Cytotoxicity Test Cytotoxicity assays were performed to evaluate the toxicity of the composition to fibroblasts and keratinocytes. The specific procedures were as follows: 8 × 10⁻⁶ mg / L of the composition was injected into each cell. 3 Each hole and 1×10 4 Human fibroblasts (Fb20081902, Guangdong Boxi Biotechnology Co., Ltd.) and human keratinocytes (HaCaT, Guangdong Boxi Biotechnology Co., Ltd.) were seeded into 96-well plates and incubated for 12 h in an incubator (37℃, 5% CO2). When the cell layering rate reached 60%, 200 μL of drug was administered per well, followed by 24 h of incubation. The experimental group received medium containing 100 g / L of the drug composition; the blank group received DMEM medium; the solvent group received DMEM medium only (no cells seeded); and the positive control group received DMEM medium containing 10 wt% DMSO. Each sample was tested in 5 replicates, and the average value was calculated. After drug administration, the cells were incubated for another 24 h, then the supernatant was discarded, and 0.5 mg / mL MTT was added and incubated at 37℃ in the dark. After 4 hours, the supernatant was aspirated, DMSO was added, and the OD value was read at 490 nm and the cell viability was calculated. Cell viability % = (OD of experimental group or positive control group - OD of solvent group) / (OD of blank group - OD of solvent group) * 100%. The results are shown in Table 5.

[0073] Table 5

[0074] As shown in Table 5, the composition provided by the present invention has weak cytotoxicity, and the cell survival rate is significantly reduced when no salmon extract-derived DNA sodium is added.

[0075] Test Example 5: Metalloproteinase Activity Inhibition Test To evaluate the inhibitory effect of the composition on MMP-1 expression, a matrix metalloproteinase expression inhibition assay was performed. The specific procedure was as follows: human fibroblasts were injected with 4 × 10⁻⁶ cells / mL of MMP-1. 4 Inoculate 12-well plates with DMEM medium at a density of 1 sample per well and incubate for 24 h in an incubator (37℃, 5% CO2). Discard the supernatant, then administer 2 mL of drug per well, followed by 24 h of incubation. The experimental group was treated with medium containing 100 g / L of the composition plus UV light (40 mJ), while the blank group was treated with DMEM medium plus UV light (40 mJ). Each sample was tested in triplicate, and the average value was calculated. Collect the supernatant and use the MMP-1 ELISA kit to detect the concentration of MMP-1 enzyme in the supernatant according to the instructions. The MMP-1 expression inhibition rate was calculated as (concentration in the blank group - concentration in the experimental group) / concentration in the blank group × 100%. The results are shown in Table 6.

[0076] Test Example 6: Elastin Expression Promotion Assay To evaluate the effect of the composition on promoting elastin expression, cell experiments were conducted to test the elastin content synthesized in human fibroblasts. The specific procedure was as follows: [Instructions for application were given]. 4 Human fibroblasts were seeded into 24-well plates at a seeding density of [number] cells / well and incubated for 12 hours in an incubator (37°C, 5% CO2). When the cell deposition rate reached 60%, 2 mL of drug was administered to each well, followed by 24 hours of incubation. The experimental group received culture medium containing 100 g / L of the drug, while the control group received DMEM medium. Each sample was tested in triplicate, and the average value was calculated. After drug administration, the plates were incubated for another 24 hours. The supernatant was collected, and the concentration of elastin in the supernatant was determined using an Elastin ELISA kit according to the manufacturer's instructions. The results are shown in Table 6.

[0077] Table 6

[0078] As shown in Table 6, the composition provided by the present invention has the effect of promoting the expression of elastin in human fibroblasts and inhibiting the expression of MMP-1 enzyme.

[0079] Test Example 7: Skin Irritation Test The skin irritation of the composition was evaluated using a closed patch test on human skin, specifically following the method for human skin patch testing as described in the "Cosmetic Safety Technical Specifications" (2015 edition). Thirty volunteers aged 25-45 years who met the test requirements were selected as subjects (half male and half female). A 300 g / L aqueous solution of the composition was added to the patch applicator, with a dosage of 0.020-0.025 mL. The blank control consisted of no added substance. The patch applicator containing the test substance was applied to the flexor side of the subject's forearm using non-irritating adhesive tape, and the skin was gently pressed with the palm of the hand to ensure even application. Skin reactions were observed 30 min (after the indentation disappeared), 24 h, and 48 h after removing the patch applicator, and the results were recorded (see Table 7).

[0080] Table 7

[0081] As can be seen from the data in Table 7, the embodiments provided by the present invention do not cause irritation to human skin or cause only mild irritation.

[0082] Application Example 1 This invention provides a firming and anti-wrinkle serum, comprising the following components by mass ratio: 25% of the composition of Example 1, 2% sodium hyaluronate, 8% glycerin, 2% propylene glycol, 0.1% p-hydroxyacetophenone, 0.5% phenoxyethanol, and 0.011% lactic acid, with the balance being water.

[0083] The preparation method of the above-mentioned firming and anti-wrinkle serum is as follows: (1) Dissolve the humectant and preservative in water at 50°C, stir well, and then cool down to 20~30°C to obtain solution B; (2) Add the firming and anti-wrinkle composition to the solution B, stir evenly, add pH adjuster and defoaming to obtain firming and anti-wrinkle essence.

[0084] Application Example 2 This invention provides a firming and anti-wrinkle serum, comprising the following components by mass ratio: 30% of the composition of Example 2, 1.5% sodium hyaluronate, 8% glycerin, 2% propylene glycol, 3% butylene glycol, 0.1% p-hydroxyacetophenone, 0.5% phenoxyethanol, 0.8% carbomer 940, and 0.011% lactic acid, with the balance being water.

[0085] The preparation method of the above-mentioned firming and anti-wrinkle serum is as follows: (1) Dissolve the thickener in water, heat to 60°C to dissolve, keep warm for later use, and obtain solution A; (2) Add humectant and preservative to solution A, stir evenly, and then cool to 20~30℃ to obtain solution B; (3) Add the firming and anti-wrinkle composition to the solution B, stir evenly, add pH adjuster and defoamer to obtain firming and anti-wrinkle essence.

[0086] Application Comparison Example 1 This invention provides a firming and anti-wrinkle serum, comprising the following components by mass ratio: 25% of the composition of Comparative Example 1, 2% sodium hyaluronate, 8% glycerin, 2% propylene glycol, 0.1% p-hydroxyacetophenone, 0.5% phenoxyethanol, and 0.011% lactic acid, with the balance being water.

[0087] The preparation method is the same as in Application Example 1.

[0088] Application Comparison Example 2 This invention provides a firming and anti-wrinkle serum, comprising the following components by mass ratio: 25% of the composition of Comparative Example 2, 2% sodium hyaluronate, 8% glycerin, 2% propylene glycol, 0.1% p-hydroxyacetophenone, 0.5% phenoxyethanol, and 0.011% lactic acid, with the balance being water.

[0089] The preparation method is the same as in Application Example 1.

[0090] Application Comparison Example 3 This invention provides a firming and anti-wrinkle serum, comprising the following components by mass ratio: 25% of the composition of Comparative Example 3, 2% sodium hyaluronate, 8% glycerin, 2% propylene glycol, 0.1% p-hydroxyacetophenone, 0.5% phenoxyethanol, and 0.011% lactic acid, with the balance being water.

[0091] The preparation method is the same as in Application Example 1.

[0092] Test Example 8: Human Anti-wrinkle Test To evaluate the firming and anti-wrinkle effect of a serum on the skin around the eyes using a human skin elasticity test, the following procedures were performed: Forty volunteers aged 35-45 with undamaged skin (half male and half female) were randomly assigned to groups of eight, each group receiving a different serum. Each participant applied 1 mL of serum to their face by hand every evening. During the test, no other cosmetics were applied to the test area (around the eyes). The serum was used continuously for four weeks. The elasticity of the skin around the eyes was measured before the test and after four weeks using an MPA580 skin elasticity tester. The rate of change in skin elasticity was calculated as [Σ(T4-T0) / T4]÷n×100%, where T4 is the elasticity value of the skin around the eyes after four weeks, T0 is the elasticity value of the skin around the eyes before the test, and n is the number of participants. The results are shown in Table 8.

[0093] Table 8

[0094] As shown in Table 8, the application examples provided by this invention have a higher skin elasticity improvement effect compared to the control examples. (See attached specification) Figure 1 The photos show the corners of the eyes of a subject who used the essence of Application Example 1 before and after the experiment. The photos show that the wrinkles around the subject's eyes became significantly shallower after the administration of the drug.

[0095] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A firming and anti-wrinkle composition, characterized in that, Based on its mass fractions, it includes the following components: 0.2~0.5 parts growth factor promoter, 1 part Sichuan pepper ( Lindera glauca The product contains fruit extract and 0.02-0.08 parts of salmon extract-derived DNA sodium; the growth factor promoter is selected from at least one of hydroxypinazone retinate, psoralen, Bidens trifoliata extract, asiaticoside, palmitoyl tripeptide-1 and palmitoyl tetrapeptide-7.

2. The firming and anti-wrinkle composition according to claim 1, characterized in that, The extraction method for the extract of Litsea cubeba fruit includes the following steps: The surfactant and water are mixed to obtain an entrainer, which is then added to a solvent tank. The mountain pepper fruits were washed, dried, and crushed to obtain mountain pepper fruit powder, which was then placed in an extraction vessel. Supercritical CO2 fluid is introduced through a solvent vessel and then into an extraction vessel for extraction reaction. Cooling and depressurization recover CO2, yielding an oil-water mixture; Preheated water was added to the oil-water mixture, and homogenization and phase inversion were performed to obtain an emulsion of Litsea cubeba fruit extract.

3. The firming and anti-wrinkle composition according to claim 2, characterized in that, The surfactant is at least one of PEG-30 dihydroxystearate, sorbitan monopalmitate, sorbitan monolaurate and polyglycerol-2-dihydroxystearate; And / or, the mass ratio of the surfactant to water is (10:90) to (3:97); And / or, the crushing refers to crushing the peony fruit to 60-80 mesh; And / or, the ratio of the mountain pepper fruit to the entrainer is 1 kg: 100~300 mL; And / or, the extraction pressure is 20~30MPa, the temperature is 40~60℃, and the supercritical CO2 flow rate is 20-30kg / h; And / or, the mixing ratio of the pepper fruit powder and supercritical CO2 is 1g: 2~4mL; And / or, the cooling and pressure reduction refers to the pressure being reduced to 5-10 MPa, and / or the temperature being reduced to 25℃-40℃; And / or, the preheated water temperature is 40~60℃; And / or, the amount of preheated water added is determined according to the water content of the pepper fruit extract being 40% to 60%; And / or, the homogenization phase inversion refers to homogenization at 800~1200 rpm.

4. The firming and anti-wrinkle composition according to any one of claims 1 to 3, characterized in that, The firming and anti-wrinkle composition also includes one or more of the following: soluble proteoglycans, serum albumin, fibronectin, collagen, chitosan, Inonotus obliquus extract, Betula alba sap, Bletilla striata root water, ceramide, and sodium polyaspartate.

5. The firming and anti-wrinkle composition according to claim 4, characterized in that, The firming and anti-wrinkle composition comprises, by weight percentage: 0.2%–3% hydroxypinazone retinate, 0.2%–4% asiaticoside, 1%–5% palmitoyl tripeptide-1, 20%–30% peperomia fruit extract and 0.5%–1% salmon extract-derived DNA sodium, with the balance being betula alba sap.

6. The firming and anti-wrinkle composition according to claim 4, characterized in that, The firming and anti-wrinkle composition comprises, by weight percentage: 0.2%–3% hydroxypinazone retinate, 1%–2% psoralen, 5%–10% Bidens trifoliata extract, 20%–30% Litsea cubeba fruit extract, 0.5%–1% salmon extract-derived sodium DNA, 2%–3% soluble proteoglycans, 0.5%–3% chitosan, and 5%–10% Inonotus obliquus extract, with the balance being water.

7. The application of the firming and anti-wrinkle composition according to any one of claims 1 to 6, characterized in that, The firming and anti-wrinkle composition is applied to serums, face creams, lotions, or masks.

8. A firming and anti-wrinkle serum, characterized in that, The serum comprises the following components by weight percentage: 10% to 30% of the firming and anti-wrinkle composition according to any one of claims 1 to 6, 7% to 15% of moisturizer, 0.3% to 1.5% of preservative, 0% to 7% of thickener and 0.01% to 0.05% of pH adjuster, with the balance being water.

9. The firming and anti-wrinkle serum according to claim 8, characterized in that, The moisturizer is selected from at least one of glycerin, butylene glycol, dipropylene glycol, propylene glycol, pentanediol, and sodium hyaluronate; And / or, the preservative is selected from at least one of p-hydroxyacetophenone, phenoxyethanol and ethylhexylglycerin; And / or, the thickener is selected from at least one of carbomer and xanthan gum; And / or, the pH adjuster is selected from at least one of citric acid, lactic acid and triethanolamine.

10. The method for preparing the firming and anti-wrinkle serum according to claim 8, characterized in that, Includes the following steps: (1) Dissolve the thickener in water, heat to 50~60℃ to dissolve, keep warm for later use, and obtain solution A; (2) Add humectant and preservative to solution A, stir evenly, and then cool to 20~30℃ to obtain solution B; (3) Add the firming and anti-wrinkle composition to the solution B, stir evenly, add pH adjuster and defoamer to obtain firming and anti-wrinkle essence.

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