Preparation method of anti-wrinkle essence

Abstract

The invention provides a preparation method of an anti-wrinkle essence, which is used for preparing the anti-wrinkle essence comprising polydeoxyribonucleotide and human collagen which are used as anti-wrinkle agents, ribonucleic acid which is used as a neurotransmitter inhibitor, and an antioxidant, a humectant, a thickening agent, a diluent, a pH regulator and a preservative which are added in cosmetics in an acceptable manner. Comprising the following steps: putting a diluent into a preparation pot, and scattering a thickening agent carbomer; the preparation method comprises the following steps: pre-dispersing a humectant sodium hyaluronate, glycerol, butanediol and sclerotium gum serving as a thickener, adding the pre-dispersed humectant sodium hyaluronate, glycerol and butanediol into a preparation pot, stirring and heating, adding pentanediol serving as the humectant and an antioxidant, continuously stirring until the materials are uniform, and cooling for the first time; adding a pH regulator into the preparation pot subjected to primary cooling to regulate the pH value, carrying out secondary cooling, adding a preservative, a neurotransmitter inhibitor, an anti-wrinkle agent and other materials serving as a thickening agent and a humectant, continuously stirring until the materials are uniform, standing, and filtering by using a screen, so as to obtain the product.

Description

Technical Field

[0001] This invention relates to a preparation method, and more particularly to a preparation method for an anti-wrinkle serum, belonging to the field of anti-wrinkle serum manufacturing technology. Background Technology

[0002] As people age, and due to factors such as genes and environment, the metabolism of human skin gradually slows down, resulting in a decrease in water and subcutaneous fat, coarsening of elastic fibers, loss of luster, and ultimately the formation of wrinkles.

[0003] Based on the principles of skin wrinkle formation, wrinkles can be divided into static wrinkles and dynamic expression lines, among which:

[0004] Static wrinkles are caused by the lack of human collagen and elastin, which leads to skin laxity, sagging, and local layering. They manifest as sagging upper facial skin, resulting in wrinkles such as lower eyelid wrinkles, nasolabial folds, cheek wrinkles, and neck wrinkles. They mainly appear around the age of forty, after which permanent static wrinkles will gradually remain on the face.

[0005] The cause of dynamic expression lines is the surface skin wrinkles caused by muscle contraction when people make facial expressions. These mainly include frown lines, forehead wrinkles, and crow's feet. In addition to the natural aging of the skin, the most important factor is the excessive contraction of facial muscles, which usually appears around the age of 20.

[0006] Wrinkles are a typical symptom of skin aging and a manifestation of the loss of skin elasticity, which has always been a concern for people.

[0007] Although skin aging is an irreversible physiological phenomenon, its process can be slowed down or delayed.

[0008] The most common way to delay or slow down the appearance of facial wrinkles is to apply anti-wrinkle cosmetics, including anti-wrinkle essential oils. Currently, there are many types of anti-wrinkle cosmetics available, such as:

[0009] Invention patent applications: "A multi-peptide anti-aging repair mask essence and its preparation method" (application number: CN201510009980.6), "A cosmetic" (application number: CN201710215761.2), "A skin care composition with repair and regeneration, moisturizing and anti-aging effects" (application number: CN201711162611.6), "Anti-wrinkle composition and anti-wrinkle cream" (application number: CN202011544525.3), and "A scorpion venom polypeptide and its use in whitening and anti-wrinkle cosmetics or drugs" (application number: CN202011544525.3). Application No.: CN202410322953.3), Invention Patent Application No.: CN202410115384.5, Invention Patent Application No.: CN202410227711.6, Invention Patent Application No.: CN202410390651.X, Invention Patent Application No.: CN202410428860.9, etc.

[0010] As can be seen from the above-listed anti-wrinkle skin products, existing anti-wrinkle skin products contain a variety of ingredients, such as extracts of various precious plants and animals, as well as ingredients such as peptides, retinol (vitamin A), xylose derivatives, and serotonin.

[0011] However, the extracts of precious plants and animals have complex compositions and their anti-skin aging effects are not certain. Furthermore, high concentrations of these extracts can cause skin intolerance issues and may clog hair follicles, damaging the skin. Retinol and other similar products can irritate the skin, while Pro-Xylane and peptides are relatively mild, but their anti-wrinkle effects are slow and not long-lasting.

[0012] In existing technologies, there are various methods for preparing serums, such as:

[0013] The invention patent application "A whitening and anti-wrinkle essence and its preparation method" (application number: CN202310027116.3) provides a technical route in which the aqueous phase is added to water, stirred and homogenized, the temperature of the mixture is controlled at 80-85°C and kept warm; then the temperature is lowered to 50-55°C, the whitening and anti-wrinkle composition dispersed with glycerin is added, and stirring is continued until homogenized; when the temperature is lowered to 40-45°C, the moisturizing phase is added, and when the temperature is lowered to 38-40°C, the preservative and fragrance are added, and the essence is obtained after stirring.

[0014] The invention patent application "Whitening and Anti-wrinkle Composition Containing Antrodia camphorata and Essence Containing the Composition and Preparation Method of Essence" (application number: CN201710850641.X) provides the following method: (1) Add the whitening and anti-wrinkle composition of Antrodia camphorata, glycerin, butylene glycol, 1,2-pentanediol, p-hydroxyacetophenone, and phenoxyethanol to a container, heat to 75°C, keep warm and stir at high speed for 20 minutes to fully disperse and dissolve the extract; (2) Add sodium hyaluronate, Lactobacillus / soy milk fermentation product filtrate and water to another container, stir to completely dissolve the materials; when (1) drops to 50°C, pour it into (2) and stir at high speed for 20 minutes to mix the liquid evenly; after vacuum degassing the solution, the preparation method of the essence is obtained.

[0015] The invention patent application "An Anti-wrinkle Essence and Its Preparation Method" (application number: CN202311383235.9) provides a method for preparing an anti-wrinkle essence by adding water, thickener, glyceryl polyether-26 (a moisturizer), and sodium hyaluronate (a moisturizer) to an emulsifying pot, homogenizing until completely dispersed, stirring, heating to 85-90°C, and maintaining the temperature for 20-30 minutes until defoaming is complete, to obtain material A; adding a skin moisturizer and oil-soluble anti-wrinkle ingredients to an oil phase pot, stirring and heating to 80-85°C until completely dissolved, to obtain material B; adding material B to material A while stirring at a uniform speed, stirring until the oil phase forms uniform oil droplets, to obtain material C; and then adding water-soluble... The anti-wrinkle ingredients, butylene glycol, 1,2-pentanediol, and hydroxyphenylpropylamide benzoic acid are mixed evenly to obtain material D; the antioxidant and moisturizer 1,3-butanediol are heated to 50-60℃ and completely dissolved to obtain material E; the pH adjuster and water are stirred at room temperature and dissolved until transparent to obtain material F; niacinamide and water are stirred at room temperature and dissolved until transparent to obtain material G; the moisturizer 1,2-hexanediol, fragrance, and solubilizer are mixed evenly to obtain material H; when material C is cooled to 42-45℃, materials D, E, F, G, and H are added, stirred until the mixture is uniform, and then cooled to below 40℃ to obtain the essence.

[0016] As can be seen from the above preparation methods, the preparation methods of essence are mainly designed based on its components, including steps such as ingredient preparation, mixing, and stirring. They may also include processes such as heating and cooling, and some may even include vacuum degassing. Obviously, these preparation methods may not be suitable for some essences containing active materials with special properties. These active materials with special properties are often the true essence of the so-called essence, and their degree of dispersion and activity retention determine the efficacy of the essence. Summary of the Invention

[0017] To meet people's desire to delay and slow down skin aging, maintain its delicate smoothness and youthful beauty, this invention provides a method for preparing an anti-wrinkle essence, so as to obtain an essence that can effectively delay the aging of facial skin, enhance its elasticity, and achieve the purpose of quickly reducing wrinkles and wrinkles while maintaining the skin's delicate smoothness for a long time.

[0018] To achieve the above objectives, the present invention provides the following technical solution:

[0019] A method for preparing an anti-wrinkle serum, wherein:

[0020] The anti-wrinkle serum comprises a clear and transparent mixture consisting of polydeoxyribonucleotides and human collagen as anti-wrinkle agents, ribonucleic acid as a neurotransmitter inhibitor, and antioxidants, moisturizers, thickeners, diluents, pH adjusters, and preservatives acceptable for addition to cosmetics. Its preparation method includes the following steps:

[0021] Place the diluent in the mixing pot and slowly sprinkle in carbomer as a thickener.

[0022] Sodium hyaluronate, glycerin, and butylene glycol, which are used as humectants, are pre-dispersed with sclerotium gum, which is used as a thickener, to prepare a uniformly dispersed mixture.

[0023] Then, the mixture is added to the preparation pot, stirred and heated, and then pentylene glycol as a humectant and an antioxidant are added. Stirring continues until the material in the preparation pot is uniform, and then the first cooling is carried out.

[0024] Add a pH adjuster to the prepared pot after the first cooling to adjust the pH, then perform a second cooling. Next, add preservatives, neurotransmitter inhibitors, anti-wrinkle agents, and other materials as thickeners and humectants. Continue stirring until homogeneous, let stand, and then filter through a sieve to obtain the final product.

[0025] Furthermore, in the above preparation method, the heating temperature is 70-75°C, the first cooling temperature is 50-55°C, and the second cooling temperature is below 40°C.

[0026] Furthermore, in the above preparation method, the stirring speed is 5-50 r / min, and the sieve used for filtration has a mesh size of 200 mesh.

[0027] Furthermore, in the above preparation method, the preparation pot is a stainless steel steam heating pot with a stirring device, and the preparation pot is sterilized by boiling water for 30 minutes, then sealed and cooled to room temperature before use.

[0028] Furthermore, in the above preparation method, the stirring device is a frame stirrer with a screen stirring frame.

[0029] Furthermore, in the anti-wrinkle serum prepared by the above method:

[0030] The antioxidant is p-hydroxyacetophenone, the diluent is water, the pH adjuster is arginine, and the preservatives are glyceryl caprylate, capryloyl hydroxamic acid, and 1,3-propanediol.

[0031] Furthermore, in the 100mL anti-wrinkle serum prepared by the above method:

[0032] The compound contains 0.50–0.52 g of the polydeoxyribonucleotide, 0.10–0.12 g of the human collagen, 0.010–0.012 g of the ribonucleic acid, 0.50–0.52 g of the p-hydroxyacetophenone, 0.05–0.06 g of the sodium hyaluronate, 6.0–6.2 g of the glycerol, 4.0–4.2 g of the butylene glycol, 3.0–3.2 g of the pentanediol, 0.10–0.12 g of the sclerotium sclerotium gum, 0.10–0.12 g of the carbomer, 0.10–0.12 g of the arginine, and a total of 0.30–0.32 g of the glyceryl caprylate, capryloyl hydroxamic acid, and 1,3-propanediol, with the remainder being water.

[0033] Furthermore, the anti-wrinkle essence prepared by the above preparation method is a clear purple to purplish-red transparent liquid with a pH value of 6.5 to 7.5 and a relative density of 1.02 to 1.04 at 20°C.

[0034] Furthermore, the anti-wrinkle essence prepared by the above preparation method, when tested according to Chapter 4 of the National Standard of the People's Republic of China "Technical Specifications for Cosmetic Safety" (2015 edition), has a mercury content ≤1mg / kg, a lead content ≤10mg / kg, an arsenic content ≤2mg / kg, and a cadmium content ≤5mg / kg.

[0035] Furthermore, the anti-wrinkle essence prepared by the above preparation method, when tested according to Chapter 5 of the National Standard of the People's Republic of China "Technical Specifications for Cosmetic Safety" (2015 edition), has a total bacterial count ≤1000 CFU / g, a total mold and yeast count ≤100 CFU / g, and no detectable thermotolerant coliforms, Staphylococcus aureus, or Pseudomonas aeruginosa.

[0036] Compared with the prior art, the outstanding beneficial effects and significant progress of the present invention are as follows:

[0037] 1) The preparation method provided by this invention can prepare an anti-wrinkle essence comprising polydeoxyribonucleotides and human collagen as anti-wrinkle agents, ribonucleic acid as a neurotransmitter inhibitor, and antioxidants, moisturizers, thickeners, diluents, pH adjusters, and preservatives that can be added to cosmetics. Among the polydeoxyribonucleotides (abbreviated as sodium DNA or PDRN) and human collagen as anti-wrinkle agents, sodium DNA or PDRN is a skin cell repair activator that can quickly activate the skin's inherent regenerative ability and activate the production of human collagen. Human collagen can regulate the internal and external balance of fibroblasts, stimulate the production of type I and III human collagen, support the epidermis and stretch the dermis, and activate skin vitality in both directions. It has a good effect of delaying skin aging and enhancing skin elasticity. As for the ribonucleic acid RNA as a neurotransmitter inhibitor, it can effectively inhibit the release of neurotransmitters and inhibit expression lines. The essence obtained by organically and ingeniously combining these functional and effective ingredients can quickly reduce wrinkles and maintain smoothness of facial skin after simple application.

[0038] 2) The anti-wrinkle essence prepared by the preparation method provided by this invention has been tested and proven to meet the requirements of the "Cosmetic Safety Technical Specifications" (2015 edition) in terms of all its physicochemical indicators. It is safe and hygienic, and no adverse reactions occur after use, thus meeting the safety standards for cosmetics.

[0039] 3) The anti-wrinkle essence prepared by the method provided by this invention involves pre-dispersing sodium hyaluronate, glycerin, and butylene glycol as moisturizers with sclerotium tumefaciens as a thickener to form a uniformly dispersed mixture. Then, the mixture is added to the preparation pot, stirred and heated, followed by the addition of pentylene glycol as a moisturizer and an antioxidant. Stirring continues until the material in the preparation pot is uniform, and the various effective ingredients are more evenly dispersed. The preparation method provided by this invention involves two cooling processes: the first cooling is within the temperature range of 50–55°C, and the second cooling is within the temperature range of below 40°C, which better preserves the performance of the various effective ingredients. In particular, the addition of a pH adjuster after the first cooling, followed by the second cooling, makes the active ingredients more stable and compatible, and the effective ingredients are more easily absorbed. Furthermore, this invention uses a sieve filter, making the product more refreshing, non-sticky, and finer. In summary, the essence prepared by this invention has uniformly dispersed active ingredients, maintains good performance, is gentle and non-irritating, refreshing and non-sticky, suitable for sensitive skin, and its active ingredients are easily absorbed, leaving the skin feeling more moisturized, stronger, more radiant, softer, and more hydrated and translucent. The skin becomes more delicate, smooth, plump, and has the soothing effects of improving rough skin, reducing facial fine lines and wrinkles, and alleviating redness, itching, and burning sensations. In particular, it has excellent anti-wrinkle effects, is simple to prepare, and convenient to use. Compared with existing technologies, it has outstanding beneficial effects and significant progress, and therefore has great promotion and application value. Attached Figure Description

[0040] To more clearly illustrate the technical solution of the present invention and the technical effects of implementing the present invention, the accompanying drawings used in the embodiments of the present invention will be briefly introduced below.

[0041] It is obvious:

[0042] The accompanying drawings described below are only some of the drawings of embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without any creative effort, but these other drawings also belong to the drawings required for the embodiments of the present invention, wherein:

[0043] Figure 1 A schematic diagram of the frame stirrer structure of the preparation pot used in the anti-wrinkle serum preparation method provided by the present invention.

[0044] Figure 2 A bar chart showing the change in the average length of crow's feet 4 hours after the application of the test substance;

[0045] Figure 3 A bar chart showing the change in the average width of crow's feet 4 hours after the application of the test substance;

[0046] Figure 4 A bar chart showing the change in the average depth of crow's feet 4 hours after the application of the test product;

[0047] Figure 5 A bar chart showing the change in the average length of crow's feet after use of the test product;

[0048] Figure 6 A bar chart showing the change in the average width of crow's feet after use of the test product;

[0049] Figure 7 A bar chart showing the change in the average depth of crow's feet after use of the test product;

[0050] Figure 8 A bar chart showing the change in the average width of nasolabial folds after the use of the test substance;

[0051] Figure 9 A bar chart showing the change in the average depth of nasolabial folds after the use of the test substance;

[0052] Figure 10 A bar chart showing the change in the average width of forehead wrinkles after use of the test product;

[0053] Figure 11 A bar chart showing the average change in forehead wrinkle depth after use of the test product;

[0054] Figure 12 A bar chart showing the average change in forehead wrinkle depth after use of the test product;

[0055] Figure 13 A bar chart showing the change in the skin elasticity parameter R2 after use of the test substance;

[0056] Figure 14 A bar chart showing the change in the skin elasticity parameter F4 after use of the test substance;

[0057] Figure 15 A bar chart showing the change in skin glossiness values ​​after use of the test substance;

[0058] Figure 16 A bar chart showing the water content of the stratum corneum of the skin;

[0059] Figure 17 A bar chart showing the changes in transepidermal water loss rate of the skin at different time points after the application of the test substance;

[0060] Figure 18 A bar chart showing changes in skin hemoglobin after use of the test substance;

[0061] Figure 19 A bar chart showing the change in skin a* values ​​after use of the test substance;

[0062] Figure 20 A set of photographs showing changes in local facial features before and after treatment was used for some of the subjects;

[0063] Figure 21 This is a photo set showing the improvement in wrinkles before and after use for some participants;

[0064] Figure 22 A photo set showing the improvement in wrinkles after 4 hours of use on some subjects.

[0065] In the picture:

[0066] 10 - Stirring shaft, 20 - Stirring frame, 30 - Screen;

[0067] p-values ​​are defined as follows: "ns" indicates no statistical difference (p > 0.05); "*" indicates a significant difference (0.01 ≤ p < 0.05); "**" indicates a highly significant difference (0.001 ≤ p < 0.01); and "***" indicates an extremely significant difference (p < 0.001). Detailed Implementation

[0068] To make the technical solution, beneficial effects and significant progress of the present invention clearer and more comprehensive, the technical solution provided by the present invention will be clearly and completely described below through specific embodiments and their effects. Obviously, all described embodiments are only some embodiments of the present invention, and not all embodiments.

[0069] Based on the embodiments and effects of this invention, all other embodiments and effects obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0070] It should be noted that the terms "firstly," "secondly," etc., in the specification, claims, and embodiments and effect examples of this invention are only used to distinguish different objects and not to describe a specific order; furthermore, the term "comprising" and any variations thereof are intended to cover non-exclusive inclusion, for example, including not only a series of listed steps or units of a process, method, system, product, or device, but also optionally steps or units not listed, or optionally other steps or units inherent to these processes, methods, products, or devices.

[0071] It should be understood that some basic operational terms commonly used in the art are used in the description of the embodiments of the present invention, such as "mixing", "stirring", "filtering", "heating", and "cooling". These terms should be interpreted broadly, that is, they can refer to conventional operations performed using various conventional equipment and instruments in the art, or they can refer to operations performed using the latest equipment, such as programmed operations and unmanned automatic operations. Unless otherwise expressly defined, those skilled in the art should understand the specific meaning of the above terms in the present invention according to the specific circumstances and adopt specific operating methods to achieve their operating objectives.

[0072] It should also be noted that the following specific embodiments can be combined with each other, and the same or similar concepts or processes may not be described again in some embodiments and their effects.

[0073] Furthermore, all the raw materials and excipients, preservatives and other reagents, production equipment and facilities involved in the following specific embodiments and effect examples are commercially available.

[0074] The technical solution of the present invention will now be described in detail with reference to specific embodiments and their effects.

[0075] Example

[0076] This embodiment provides a method for preparing an anti-wrinkle serum for quickly reducing wrinkles and maintaining long-lasting smoothness of facial skin.

[0077] A method for preparing an anti-wrinkle serum, wherein:

[0078] The anti-wrinkle serum comprises a clear and transparent mixture consisting of polydeoxyribonucleotides and human collagen as anti-wrinkle agents, ribonucleic acid as a neurotransmitter inhibitor, and antioxidants, moisturizers, thickeners, diluents, pH adjusters, and preservatives acceptable for addition to cosmetics. Its preparation method includes the following steps:

[0079] Place the diluent in a mixing pot and slowly sprinkle in carbomer as a thickener. Pre-disperse sodium hyaluronate, glycerin, and butylene glycol as humectants with sclerotium tumefaciens as a thickener to form a uniformly dispersed mixture. Then, add this mixture to the mixing pot, stir and heat. Add pentylene glycol as a humectant and an antioxidant, and continue stirring until the material in the mixing pot is uniform. Perform a first cooling. Add a pH adjuster to the mixing pot after the first cooling to adjust the pH, and perform a second cooling. Then, add preservatives, neurotransmitter inhibitors, anti-wrinkle agents, and other materials as thickeners and humectants. Continue stirring until uniform, let stand, and then filter through a sieve to obtain the final product.

[0080] Furthermore, in the above preparation method, the heating temperature is 70-75°C, the first cooling temperature is 50-55°C, and the second cooling temperature is below 40°C, so as to ensure the activity of the active ingredients under these mild conditions.

[0081] Furthermore, in the above preparation method, the stirring speed is 5-50 r / min, and the filtration screen is 200 mesh, so as to fully disperse the active ingredients and effectively remove particulate matter and entrained air bubbles from the essence.

[0082] Furthermore, in the above preparation method, the preparation pot is a stainless steel steam heating pot with a stirring device, and the preparation pot is sterilized by boiling water for 30 minutes, then sealed and cooled to room temperature before use, so as to ensure the hygiene and safety of the product.

[0083] Furthermore, such as Figure 1 The schematic diagram of the frame stirrer structure of the preparation pot used in the anti-wrinkle essence preparation method provided by the present invention is shown. The stirring device is a frame stirrer including a stirring shaft 10, a stirring frame 20 and a sieve 30.

[0084] Furthermore, in the anti-wrinkle serum prepared by the above preparation method, the antioxidant is p-hydroxyacetophenone, the diluent is water, the pH adjuster is arginine, and the preservatives are glyceryl caprylate, capryloyl hydroxamic acid, and 1,3-propanediol.

[0085] Furthermore, in the 100mL anti-wrinkle serum prepared by the above method:

[0086] It contains 0.50–0.52 g of polydeoxyribonucleotides, 0.10–0.12 g of human collagen, 0.010–0.012 g of ribonucleic acid, 0.50–0.52 g of p-hydroxyacetophenone, 0.05–0.06 g of sodium hyaluronate, 6.0–6.2 g of glycerol, 4.0–4.2 g of butylene glycol, 3.0–3.2 g of pentylene glycol, 0.10–0.12 g of sclerotium tumefaciens gum, 0.10–0.12 g of carbomer, 0.10–0.12 g of arginine, and 0.30–0.32 g of a combination of glyceryl caprylate, capryloyl hydroxamic acid, and 1,3-propanediol. The remainder is water.

[0087] Furthermore, the anti-wrinkle essence prepared by the above preparation method is a clear purple to purplish-red transparent liquid with a pH value of 6.5 to 7.5 and a relative density of 1.02 to 1.04 at 20°C.

[0088] Further:

[0089] The anti-wrinkle serum prepared by the above method, when tested according to Chapter 4 of the National Standard of the People's Republic of China "Technical Specifications for Cosmetic Safety" (2015 edition), has the following contents: mercury ≤1mg / kg, lead ≤10mg / kg, arsenic ≤2mg / kg, and cadmium ≤5mg / kg.

[0090] The anti-wrinkle essence prepared by the above method, when tested according to Chapter 5 of the National Standard of the People's Republic of China "Technical Specifications for Cosmetic Safety" (2015 edition), has a total bacterial count ≤1000 CFU / g, a total mold and yeast count ≤100 CFU / g, and no detectable thermotolerant coliforms, Staphylococcus aureus, or Pseudomonas aeruginosa.

[0091] From the above description, it can be seen that:

[0092] The preparation method provided in this embodiment can prepare an anti-wrinkle serum comprising polydeoxyribonucleotides and human collagen as anti-wrinkle agents, ribonucleic acid as a neurotransmitter inhibitor, and antioxidants, moisturizers, thickeners, diluents, pH adjusters, and preservatives acceptable for addition to cosmetics, wherein:

[0093] Polydeoxyribonucleotides, also known as sodium DNA or PDRN, which are anti-wrinkle agents, are skin cell repair activators that can quickly activate the skin's inherent regenerative capacity and activate the production of human collagen.

[0094] Human collagen, which is also an anti-wrinkle agent, can regulate the internal and external balance of fibroblasts, stimulate the production of type I and III human collagen, support the epidermis and stretch the dermis, activate skin vitality in both directions, and has a good effect of delaying skin aging and enhancing skin elasticity.

[0095] RNA, as a neurotransmitter inhibitor, can effectively inhibit the release of neurotransmitters, thereby inhibiting the formation of expression lines.

[0096] Experiments have shown that:

[0097] The anti-wrinkle essence prepared by the above-mentioned anti-wrinkle agents, neurotransmitter inhibitors, antioxidants, moisturizers, thickeners, diluents, pH adjusters and preservatives, especially the organic and ingenious combination of sodium DNA and human collagen as anti-wrinkle agents and RNA as neurotransmitter inhibitors, can quickly reduce wrinkles and maintain smoothness of the skin after being simply applied to the face, allowing the user to regain a youthful and beautiful appearance.

[0098] Meanwhile, since the above-mentioned anti-wrinkle essence was prepared using suitable equipment and processes, the anti-wrinkle essence prepared was tested and tested, and its various physicochemical indicators all met the requirements of the "Cosmetic Safety Technical Specifications" (2015 edition). It is safe and hygienic, and no adverse reactions occurred after use, thus meeting the safety standards for cosmetics.

[0099] As can be seen, the anti-wrinkle essence prepared by the preparation method provided in this embodiment has uniformly dispersed active ingredients and maintains good performance. As a result, the prepared anti-wrinkle essence is gentle and non-irritating, refreshing and non-sticky, suitable for sensitive skin. The active ingredients are easily absorbed, and the skin feels more moisturized, stronger, more radiant, softer, and more hydrated and translucent. The skin becomes more delicate, smooth, plump, and has the soothing effects of improving rough skin, fading facial fine lines and wrinkles, and reducing redness, itching, and burning sensations. In particular, it has excellent anti-wrinkle effects, is simple to prepare, and convenient to use. Compared with the prior art, it has outstanding beneficial effects and significant progress, and therefore has great promotion and application value.

[0100] To further help understand the technical effects of the anti-wrinkle serum provided in this embodiment, the following will further explain the above technical solution through the formulation examples of three specific production batches of anti-wrinkle serum listed in Table 1.

[0101] Note: In Table 1, 240124F61, 240201F55, and 240226F60 are three batches of anti-wrinkle serums. "Content in 100mL (g)" refers to the weight of the constituent substances contained in 100mL of anti-wrinkle serum.

[0102] Table 1

[0103]

[0104]

[0105] Polydeoxyribonucleotides (PDRN) derived from salmon with a molecular weight ≤1000kDa were purchased from Ruijiming (Shandong) Biotechnology Co., Ltd., etc.; human collagen, type I, type III, or a combination of both, was purchased from Jiangsu Chuangjian Medical Technology Co., Ltd., etc.; ribonucleic acid (RNA) was purchased from Shaanxi Harvard Biotechnology Co., Ltd., etc.; p-hydroxyacetophenone and 1,2-pentanediol were from Symrise Fragrance & Flavor (Nantong) Co., Ltd., etc.; sodium hyaluronate was from Bloomage Biotechnology Co., Ltd., etc.; glycerol was from Tyco Brown Chemical (Zhangjiagang) Co., Ltd., etc.; carbomer was from Lubrizol Advanced Materials, Inc.; butylene glycol was from KH NeoChem CO., Ltd., etc.; 1,3-propanediol was from Primient Covation LLC, etc.; sclerotium sclerotium gum was from ALBAN MULLER INTERNATIONAL, SAS, Cargill France SAS, etc.; arginine was from Ajinomoto "L-arginine" in Japan, etc.; glyceryl caprylate was purchased from INOLEX, Inc., etc.

[0106] To further help understand the technical effects that the technical solution provided in this embodiment can achieve, the following specific examples will be used to further illustrate the efficacy of the anti-wrinkle essence prepared by the preparation method provided in the above embodiment.

[0107] Example 1: This example evaluates the safety of the anti-wrinkle serum by testing its physicochemical properties using the preparation method provided in the above embodiments.

[0108] 1) The purple to purplish-red transparent anti-wrinkle serum liquid of batch number 240226F60 provided in the above embodiments was selected for physicochemical index testing. The relevant test items, test methods, test standards and test results are listed in Table 1.1:

[0109] Table 1.1

[0110]

[0111]

[0112] 2) The anti-wrinkle serum with batch number 240201F55, which is a purple to purplish-red transparent liquid, provided in the above examples, was subjected to physicochemical index testing. The testing was conducted in accordance with the "Cosmetic Safety Technical Specifications" (2015 edition). The relevant microbiological test results are shown in Table 1.2, and the relevant physicochemical test results are shown in Table 1.3.

[0113] Table 1.2

[0114] Inspection items unit Test results Limit Total bacterial count CFU / g 150 ≤1000 Total number of molds and yeasts CFU / g <10 ≤100 Thermoresistant coliforms / g Not detected Not detectable Staphylococcus aureus / g Not detected Not detectable Pseudomonas aeruginosa / g Not detected Not detectable

[0115] Table 1.3

[0116]

[0117]

[0118] The data listed in Tables 1.1, 1.2, and 1.3 show that:

[0119] Microbiological and physicochemical tests were conducted on anti-wrinkle serums in batches 240226F60 and 240201F55. All indicators met the requirements of the National Standard of the People's Republic of China "Technical Specifications for Cosmetic Safety" (2015 edition), and thus met the safety requirements for cosmetics.

[0120] It should be noted that:

[0121] Although Example 1 only tested anti-wrinkle serums batches 240226F60 and 240201F55 for microbiological and physicochemical items and concluded that they met the safety requirements for cosmetics, in fact, the same tests can be performed on other batches of anti-wrinkle serums obtained according to the technical solution provided in the above embodiments, and the same conclusion of meeting the safety requirements for cosmetics can be drawn. For the sake of simplicity, this specification will not list more relevant test data.

[0122] Example 2: This example evaluates the effectiveness of the anti-wrinkle serum (batch number 240124F61) prepared according to the method described in the above embodiments by conducting a questionnaire survey among users.

[0123] Note: The product described in this effect example is the purple to purplish-red transparent anti-wrinkle serum liquid with batch number 240124F61 provided in the above embodiment.

[0124] Testing items: Suitable for sensitive skin, anti-wrinkle, firming, soothing, radiance, and moisturizing effects (consumer questionnaire test).

[0125] Test basis: "Cosmetic Safety Technical Specifications" (2015 edition) Human safety trial test, "Cosmetic Consumer Use Test - Questionnaire Survey Evaluation Guide" T / TDCA006-2022, Cosmetic Efficacy Evaluation (XⅢ) - Consumer Use Test [J], Daily Chemical Industry, 2021, 51(6): 6. "Evaluation of Skin Care Product Suitability for Sensitive Skin: Facial Lactic Acid Stinging Method" T / SPMA006-2023.

[0126] I. Materials and Methods

[0127] 1. Test substance: Anti-wrinkle serum (batch number: 240124F61).

[0128] 2. Purpose of the test: Using a single-blind study, this study evaluates the changes in skin wrinkles, skin firmness, skin moisture, and skin redness in 30 subjects with sensitive skin and wrinkles before and after using the test sample, through clinical physician assessment and consumer use testing. This will help to evaluate the test sample's effects on "suitability for sensitive skin", "anti-wrinkle", "firming", "soothing", "radiance", and "moisturizing".

[0129] 3. Subjects: 30 subjects were selected who met the testing criteria. A total of 30 subjects actually completed the test. There were 0 males and 30 females. Their ages ranged from 34 to 58 years, with an average age of 47.9 ± 6.1 years. See Table 2.1 for subject information.

[0130] Table 2.1 Subject Information Table

[0131]

[0132]

[0133] 4. Subject Requirements

[0134] 4.1 Inclusion criteria:

[0135] All 30 enrolled participants completed the test. Their mean age was 47.9 ± 6.1 years. All participants who completed the test met the following inclusion criteria:

[0136] The inclusion criteria for subjects were: volunteers aged 18 to 60 who met the requirements of the experiment were selected as subjects, and subjects who had a positive lactic acid stinging test or were identified as having sensitive skin according to the sensitive skin questionnaire;

[0137] Individuals with the following conditions should not be selected as subjects:

[0138] 1) Those who have used antihistamines in the past week or immunosuppressants in the past month;

[0139] 2) Those who have used any anti-inflammatory drugs at the test site within the past two months;

[0140] 3) Subjects with clinically unhealed inflammatory skin diseases;

[0141] 4) Patients with insulin-dependent diabetes mellitus:

[0142] 5) Patients with asthma or other chronic respiratory diseases who are currently receiving treatment;

[0143] 6) Individuals who have received anti-cancer chemotherapy within the past 6 months;

[0144] 7) Patients with immunodeficiency or autoimmune diseases;

[0145] 8) Breastfeeding or pregnant women;

[0146] 9) Patients who have undergone bilateral mastectomy and bilateral axillary lymph node dissection;

[0147] 10) Those whose facial scars, pigmentation, atrophy, port-wine stains, or other blemishes affect the determination of test results;

[0148] 11) Participants in other clinical trials;

[0149] 12) Individuals with highly sensitive constitutions;

[0150] 13) Non-volunteer participants or those who cannot complete the prescribed content according to the test requirements.

[0151] 4.2 Subject Restrictions:

[0152] During the trial, the test site should use the test product provided by the testing institution, discontinue the use of other products with similar effects, and maintain the usual daily product usage habits.

[0153] II. Testing Process

[0154] 1. Information registration (verification of identity information);

[0155] 2. A lactic acid stinging test will be conducted 7 to 3 days before the start of the long-term use trial (safety evaluation) to screen out those who test positive for lactic acid stinging to participate in the trial.

[0156] 3. On the day of the formal test, qualified subjects will be screened, and informed consent forms will be signed before the formal test begins.

[0157] First, the subjects washed their faces as instructed and rested for 30 minutes in an environment with a temperature of 21±1℃ and a relative humidity of 50±10%RH.

[0158] Subsequently, under the supervision of the experimenters, the product was used correctly to complete a single product use test, and the degree of discomfort such as burning, stinging, and itching on the skin after the subject's first use of the product was evaluated.

[0159] After the trial period, the experimenters conducted a lactic acid stinging test and recorded the subjects' lactic acid stinging scores and total scores at 30s, 2, 5min and 5min respectively as the baseline values ​​for lactic acid stinging scores.

[0160] After all procedures were completed, the subjects left the laboratory, used the product at home, and were followed up in the second week.

[0161] 4. Subjects used the product at home. During the follow-up visit two weeks after product use, they washed their faces as required and rested for 30 minutes in an environment with a temperature of 21±1℃ and a relative humidity of 50±10%RH. Laboratory experts evaluated the subjects' facial skin according to the safety evaluation criteria for human trial. At the same time, they completed a consumer usage questionnaire.

[0162] After the above tests were completed, the researchers conducted another lactic acid stinging test to evaluate the product and recorded the lactic acid stinging score of the skin after 2 weeks of product use.

[0163] 5. Conduct data analysis after the test is completed.

[0164] III. Test Results

[0165] 1. Facial sensation score of subjects after their first use of the product

[0166] The facial sensation scores of the subjects after their first use of the product are shown in Table 2.2.

[0167] Table 2.2 Subjects' sensory scores after first product use

[0168]

[0169] Scoring criteria:

[0170] Burning rating: 0 points: none; 1 point: mild, occasionally felt; 2 points: moderate, noticeable burning, but tolerable; 3 points: severe, very noticeable burning, requires cool water to relieve.

[0171] Stinging pain rating: 0 points: none; 1 point: mild, occasionally felt; 2 points: moderate, obvious stinging pain, but tolerable; 3 points: severe, very obvious stinging pain, requiring topical medication for relief.

[0172] Itching score: 1 point (0 degrees): almost no itching; 2 points (mild, grade I): slight itching, relieved by stroking and patting without scratching, no secondary rash caused by scratching; 3 points (moderate, grade II): obvious itching, irresistible to scratch, mild secondary rash caused by scratching is visible; 4 points and above (severe, grade III and very severe, grade IV): severe itching, affecting sleep and work, obvious secondary rash caused by scratching is visible locally; very severe itching, affecting sleep and work, emotional distress, severe and numerous secondary rashes.

[0173] 2. Skin reactions during product use by test subjects

[0174] The results of the clinical assessment of adverse skin reactions during the use of the product by laboratory personnel are summarized in Table 2.3, which summarizes the results of the cosmetic sensitivity skin suitability test.

[0175] Table 2.3 Summary of Cosmetic Suitability Test Results for Sensitive Skin

[0176]

[0177] The skin adverse reaction scoring criteria are shown in Table 2.4.

[0178] Table 2.4 Skin Adverse Reaction Scoring Criteria

[0179]

[0180]

[0181] 3. Evaluation of lactic acid stinging and self-assessment by subjects

[0182] 3.1 Evaluation of lactic acid stinging

[0183] Before and after two weeks of product use, testers conducted lactic acid stinging tests on 30 subjects. The stinging sensation at the test site was evaluated at 30s, 2, 5min and 5min, and the total sensory score was calculated. The improvement of skin irritation after product use was also evaluated. The improvement scores of stinging symptoms after product use are shown in Table 2.5.

[0184] Table 2.5 Subjects' lactic acid stinging test symptom improvement scores

[0185] Stinging Score Before Use Stinging sensation after use (total score) 3.34±0.48 1.76±0.74

[0186] 3.2 Subjective assessment of improvement in subjects' skin condition

[0187] Before and two weeks after product use, clinicians clinically assessed the symptoms of stinging, itching, burning, redness, and dryness on the skin of 30 subjects, and evaluated the improvement of skin irritation after product use. The clinical symptom improvement scores of the subjects after product use are shown in Table 2.6 below.

[0188] Table 2.6 Assessment Table of Clinical Skin Symptoms of Subjects [Total, (Minimum, Maximum)]

[0189] project Before use After use Scorching 5，(0，2) 3，(0，1) stinging 3，(0，2) 1，(0，1) itching 3，(0，1) 1，(0，1) dry 4，(0，1) 0，(0，0) erythema 3，(0，2) 1，(0，1) Desquamation 4，(0，1) 1、(0，1) Total Score 22 7

[0190] Note: The scoring criteria are 0 = no response; 1 = mild; 2 = moderate; 3 = severe.

[0191] 4. Results of the consumer questionnaire used by the test subjects

[0192] After the subjects used the product, they evaluated its effects on "suitability for sensitive skin", moisturizing, soothing, firming, and anti-wrinkle through a consumer use questionnaire. The following is the content of the questionnaire.

[0193] 4.1 The product is gentle (non-irritating) and suitable for sensitive skin.

[0194] During the product use period, all subjects experienced no adverse reactions or discomfort. The results are shown in Table 2.7 below.

[0195] Table 2.7 Results of the survey on the product's suitability for sensitive skin

[0196]

[0197]

[0198] 4.2 Skin feel after product use

[0199] After the participants finished using the product, they were asked to fill in their overall feelings about the product and their skin satisfaction after using the product. The results are shown in Table 2.8.

[0200] Table 2.8 Results of the survey on skin feel after using the product

[0201] serial number question agree p-value 1 The product has a refreshing and non-sticky texture. 100％ <0.001 2 The product is easily absorbed after use. 100％ <0.001

[0202] 4.3 Evaluation of product moisturizing and gloss effects

[0203] A consumer questionnaire survey was conducted after two weeks of product use. Participants were asked to evaluate the product's moisturizing and radiance effects based on their experience using the product. The results are shown in Table 2.9.

[0204] Table 2.9 Survey results of respondents' opinions on product claims regarding moisturizing and gloss enhancement.

[0205]

[0206] 4.4 Evaluation of the product's anti-wrinkle efficacy

[0207] Two weeks after the participants used the product, a consumer questionnaire was conducted, in which participants were asked to evaluate the product's anti-wrinkle effects on improving fine lines and texture based on their experience using the product. The results are shown in Table 2.10.

[0208] Table 2.10 Survey Results on the Anti-wrinkle Efficacy Claims of the Products by the Subjects

[0209]

[0210]

[0211] 4.5 Evaluation of Product Firming Efficacy

[0212] A consumer questionnaire survey was conducted on the subjects after they used the product for 2 weeks. The subjects were asked to evaluate the product’s skin-tightening efficacy based on their experience using the product. The results are shown in Table 2.11.

[0213] Table 2.11 Survey Results on Product Firming Efficacy Claims by Subjects

[0214]

[0215] 4.6 Evaluation of Product's Soothing Efficacy

[0216] Two weeks after the participants used the product, a consumer questionnaire was conducted, in which participants were asked to evaluate the product's soothing effects based on their experience. The results are shown in Table 2.12, which reflects the participants' assessment of the product's claims on soothing and firming effects.

[0217] Table 2.12 Survey Results of Subjects' Feedback on Product Claims of Soothing and Firming Efficacy

[0218]

[0219] IV. Test Results:

[0220] By observing 30 subjects with sensitive skin, and after using the test sample continuously for 2 weeks under normal conditions, the potential for adverse skin reactions in humans was evaluated.

[0221] Clinical evaluation by dermatologists showed that none of the 30 subjects experienced adverse reactions after use, and the subjects' self-reported results indicated that they did not feel any discomfort during or after use.

[0222] Test results show that the anti-wrinkle serum with batch number 240124F61 has high safety, is "mild (non-irritating)", and meets the requirement of "suitable for sensitive skin".

[0223] After two weeks of continuous use of the test samples under normal conditions, this consumer questionnaire test showed that 100% of the participants felt that the product was "refreshing and non-sticky" and "easily absorbed" in terms of the product's texture and feel.

[0224] After two weeks of product use, based on the fact that 100% of the participants felt that "after using the product, the skin did not feel dry or tight, but felt more moisturized", 96.7% of the participants felt that "after using the product, the skin felt more hydrated and moisturized", 93.3% of the participants felt that "after using the product, the skin felt stronger", 90.0% of the participants felt that "after using the product, the skin felt more radiant", 90.0% of the participants felt that "after using the product, the skin felt softer", and 93.3% of the participants felt that "after using the product, the skin felt more hydrated and translucent", and the results showed significant differences, it can be concluded that the product improves skin radiance after use, and therefore, it has a "moisturizing" effect.

[0225] After two weeks of product use, 93.3% of participants reported that their skin felt smoother and more delicate, 86.7% reported that their skin felt plumper and fuller, 86.7% reported that the product improved roughness, 86.7% reported that the product reduced fine lines and wrinkles, 90.0% reported that the product improved facial wrinkles, 83.3% reported that the product improved crow's feet, and 83.3% reported that the product improved under-eye wrinkles. The product showed significant differences in efficacy, with 90.0% of respondents believing it improved fine lines on the forehead, 83.3% believing it improved forehead wrinkles, 83.3% believing it improved nasolabial folds, 86.7% believing it improved frown lines, 90.0% believing it improved marionette lines (corner of the mouth lines), and 93.3% believing it improved nasal wrinkles (bridge of the nose). Therefore, it can be concluded that the product has an "anti-wrinkle" effect.

[0226] After two weeks of product use, based on the fact that 93.3% of the subjects felt that "the skin felt firmer after using the product", 90.0% of the subjects felt that "the skin felt more elastic after using the product", and 93.3% of the subjects felt that "the product improved skin laxity after using the product", and the results showed significant differences, it can be concluded that the product has a "firming" effect.

[0227] After two weeks of product use, based on the fact that 90.0% of the subjects believed that "the product effectively improved skin redness", 93.3% of the subjects believed that "the product effectively improved skin itching", and 93.3% of the subjects believed that "the product effectively improved skin burning", and the results showed significant differences, it can be concluded that the product has a "soothing" effect.

[0228] Example 3: In this example, the anti-wrinkle serum with batch number 240124F61 prepared by the method provided in the above embodiments was subjected to a human skin patch test to further evaluate its product safety.

[0229] Test item: Human skin patch test.

[0230] Test basis: Human skin patch test of the "Cosmetic Safety Technical Specifications" (2015 edition).

[0231] I. Materials and Methods

[0232] 1. Test substance: The purple to purplish-red transparent anti-wrinkle essence liquid with batch number 240124F61 obtained in the above examples.

[0233] 2. Negative control: Blank filter paper control.

[0234] 3. Subjects: A total of 30 subjects, including 7 males and 23 females, aged 19 to 45 years, with an average age of 37.0 ± 7.1 years, who met the subject voluntary inclusion criteria.

[0235] 4. Patch Test Method: Using the closed patch test method, approximately 0.025 mL of the test substance was placed in the patch tester, and a hypoallergenic adhesive tape was applied to the flexor side of the subject's forearm. The test substance was removed after 24 hours, and the skin reaction was observed at 0.5, 24, and 48 hours after removal. The results were recorded according to the skin reaction grading standard in Chapter 7 of the "Cosmetic Safety Technical Specifications" (2015 edition).

[0236] II. Test Results

[0237] The skin reactions of the 30 subjects at different observation times are shown in Tables 3.1 and 3.2.

[0238] Table 3.1 Skin reactions of 30 subjects at different observation times

[0239]

[0240]

[0241] Table 3.2 Summary of Cosmetic Human Skin Patch Test Results

[0242]

[0243] Note: Skin reaction rating levels:

[0244] 0 = negative reaction;

[0245] 1 = Suspicious reaction, only slight erythema;

[0246] 2 = Weak positive reaction (erythema reaction); erythema, infiltration, edema, and papules may be present;

[0247] 3 = Strong positive reaction (herpes reaction); erythema, infiltration, edema, papules, vesicles; the reaction may extend beyond the tested area;

[0248] 4 = Extremely strong positive reaction (confluent herpes reaction); obvious erythema, severe infiltration, edema, confluent herpes; reaction extends beyond the test area.

[0249] III. Test Results:

[0250] Human skin patch test results showed that 0 out of 30 people experienced adverse skin reactions, indicating that the anti-wrinkle essence with batch number 240124F61 obtained by the preparation method provided in the above embodiments is safe for human skin.

[0251] Example 4: This example demonstrates the effectiveness of the anti-wrinkle serum (batch number 240124F61) provided in the above embodiments, applied to the face of a subject to evaluate its suitability for sensitive skin, moisturizing, firming, anti-wrinkle, repairing, and radiance-enhancing functions.

[0252] I. Methods and Materials

[0253] 1. Testing method:

[0254] 1.1 Referring to the "Human Evaluation Method for the Repairing Efficacy of Cosmetics T / GDCA009-2022" and the "Test Method for Seven Efficacy Items of Cosmetics (Anti-wrinkle, Firming, Moisturizing, Oil Control, Repairing, Nourishing, and Soothing) T / CAB 0152-2022", the moisturizing and repairing efficacy of the product was evaluated by assessing the improvement of the main parameter transepidermal water loss rate and the secondary parameters skin hemoglobin and skin moisture content before and after the subjects used the test sample.

[0255] 1.2. Referring to the "Test Method for Seven Efficacy Items of Cosmetics (Anti-wrinkle, Firming, Moisturizing, Oil Control, Repairing, Nourishing, and Soothing) T / CAB 0152-2022" and the "Test Method for Anti-wrinkle Efficacy of Cosmetics T / ZHCA006-2019", the changes in the average length, average width, and average depth of crow's feet, forehead wrinkles, and nasolabial folds before and after subjects used the test sample were evaluated using instruments. The changes in wrinkle grading of subjects were also evaluated through clinical assessment to evaluate the anti-wrinkle efficacy of the product.

[0256] 1.3. Referring to the "Test Method for Seven Efficacy Items of Cosmetics (Anti-wrinkle, Firming, Moisturizing, Oil Control, Repairing, Nourishing, and Soothing) T / CAB 0152-2022" and the "Test Method for Firming Efficacy of Cosmetics T / TDCA003-2021", the changes in firming parameters of subjects before and after using the test sample were evaluated by instruments to assess the firming efficacy of the product.

[0257] 2. Test substance: The purple to purplish-red transparent anti-wrinkle essence liquid with batch number 240124F61 provided in the above examples.

[0258] 3. Participants: Thirty female participants who met the testing criteria were selected. None of them withdrew from the trial for personal reasons, and a total of 30 participants actually completed the test. Their ages ranged from 36 to 60 years, with a mean age of 49.7 ± 8.0 years. Participant information is shown in Table 4.1.

[0259] Table 4.1 Subject Information Table

[0260]

[0261]

[0262] 4. Subject Requirements

[0263] 4.1 Inclusion criteria: All 30 subjects enrolled completed the test, with a mean age of 49.7 ± 8.0 years. All subjects who completed the test met the following inclusion criteria.

[0264] Subject inclusion criteria:

[0265] 1) Female, aged 35 to 60, in good health;

[0266] 2) The subjects did not have serious chronic wasting diseases (such as asthma, diabetes, etc.);

[0267] 3) Able to cooperate well with researchers according to the requirements of the experimental protocol;

[0268] 4) Individuals who have not participated in other clinical trials in the past month;

[0269] 5) Currently not participating in other clinical trial projects;

[0270] 6) The subject's facial features were loose and the tested area had obvious wrinkles.

[0271] Individuals with the following conditions should not be selected as subjects:

[0272] 1) Pregnant or breastfeeding women

[0273] 2) Those who have used antihistamines within the past week or immunosuppressants within the past month;

[0274] 3) Those who have used any anti-inflammatory drugs at the test site within the past two months;

[0275] 4) Those with facial eczema, severe seborrheic dermatitis, or folliculitis;

[0276] 5) The test area has large areas of birthmarks, scratches, vitiligo, pigmented nevi, keloids, or other skin characteristics that may affect the test.

[0277] 6) Has been vaccinated in the past month or plans to be vaccinated during the study period;

[0278] 7) Patients with insulin-dependent diabetes mellitus, or patients with immunodeficiency or autoimmune diseases;

[0279] 8) Unable to cooperate with the test subject.

[0280] Subjects should also pay attention to the following:

[0281] 1) Subjects are prohibited from using other types of anti-wrinkle cosmetics during the testing period. Other products such as toners, moisturizers or other skin care products can be used as usual.

[0282] 2) Subjects are prohibited from ingesting preparations that may affect test results (such as anti-inflammatory drugs, cold medicines, and injectable medications);

[0283] 3) During the testing period, cosmetic procedures or treatments that may affect the test results, such as laser spot removal and skin rejuvenation, are prohibited.

[0284] 4) Subjects are not allowed to change any facial products or skincare habits during the test.

[0285] 4.2 Subject Restrictions:

[0286] During the test, the test site should be treated with the test product provided by the testing institution; no other products with equivalent efficacy or that may affect the test results should be used. Usage habits for other daily necessities such as soap and shower gel must remain consistent.

[0287] II. Evaluation Instruments

[0288] 1. Vapometer, test parameter: transepidermal water loss (TEWL) value of the skin. A decrease in this value indicates that the skin barrier function has been improved.

[0289] 2. Skin color measuring instrument SKINCOLORCATCH, test parameters: a* represents the red and green color of the object: positive value indicates red, negative value indicates green, and a smaller a* indicates that the red color is weakened.

[0290] 3. The German CK company MPA580 skin testing instrument - Corneometer probe for skin moisture testing. Test parameters: Corneometer value (CU) of skin stratum corneum moisture content. The higher the value, the higher the moisture content of the skin stratum corneum.

[0291] 4. The MPA580 Cutometer, a skin elasticity tester from the German company CK, has the following test parameters: skin firmness parameter F4. The smaller the value, the firmer the skin.

[0292] 5. SkinLosteester Gloss Tester. Test parameter: Skin gloss value. The higher the value, the better the skin gloss.

[0293] 6. CK MPA580 skin testing instrument from Germany - MX18 probe for testing skin melanin and hemoglobin. Test parameter: Hemoglobin. A decrease in this value indicates an improvement in skin redness.

[0294] 7. Antera3D, test parameters: skin wrinkle length, average width, average depth, etc. A decrease in this value indicates an effect of improving skin wrinkles.

[0295] 8. VISIA-CR acquires facial images of the subject.

[0296] III. Testing Process

[0297] 1. Information registration (verification of identity information).

[0298] 2. Select qualified subjects and obtain informed consent. Select subjects with sensitive skin through questionnaires. Subjects who feel that their skin barrier function is impaired, such as dryness, peeling, and redness, as well as those with obvious wrinkles around the eyes, nasolabial folds, and forehead wrinkles are selected for this test.

[0299] 3. Eligible subjects will enter the trial. Before the test, they will wash their faces with a standardized facial cleanser and then equilibrate in an environment with an ambient temperature of 20-22°C and a relative humidity of 40-60% for at least 20 minutes. Subsequently, the subjects will have their full-face images captured, and their crow's feet, forehead wrinkles, and nasolabial folds will be photographed. Then, the subjects will be tested for transepidermal water loss rate, skin moisture content, skin hemoglobin, skin firmness parameters, and initial values ​​of skin radiance.

[0300] 4. After the initial value test is completed, the laboratory technician will distribute test samples to the enrolled subjects and instruct them on how to use the product as required.

[0301] Four hours later, images of the wrinkles around the eyes of the subjects were collected again to compare the improvement of wrinkles around the eyes after four hours of use.

[0302] After the first day of testing was completed, the participants took the product away and used it at home. Throughout the testing period, the participants strictly followed the usage instructions and used the product correctly and continuously.

[0303] 5. Subjects used the product at home and were followed up in weeks 1, 2 and 4 for full-face image acquisition, photography of the crow's feet, forehead wrinkles and nasolabial folds, and repeated tests of transepidermal water loss rate, skin moisture content, skin hemoglobin, skin luster and skin firmness.

[0304] 6. After the test is completed, conduct data analysis.

[0305] IV. Test Results

[0306] 1. Improvement in crow's feet 4 hours after application of the test substance

[0307] On the day of the test, four hours after using the product, wrinkles in a fixed selected area around the corners of the subjects' eyes were analyzed.

[0308] The test results are as follows:

[0309] 1) Table 4.2 Results of length, average width, and average depth of crow's feet 4 hours after product application;

[0310] 2) Table 4.3 Results of the difference in crow's feet wrinkles 4 hours after application of the test substance;

[0311] as well as:

[0312] Figure 2 Bar chart showing the change in the average length of crow's feet 4 hours after application of the test substance;

[0313] Figure 3 Bar chart showing the change in the average width of crow's feet 4 hours after application of the test substance;

[0314] Figure 4 A bar chart showing the change in the average depth of crow's feet 4 hours after the use of the test substance.

[0315] Table 4.2 Results of length, average width, and average depth of crow's feet 4 hours after product application.

[0316]

[0317]

[0318] Table 4.3 Results of the difference in crow's feet wrinkles 4 hours after application of the test substance

[0319] Evaluation indicators 4-hour improvement 4-hour improvement rate (%) Average length of crow's feet wrinkles (mm) <![CDATA[-6.17 ＊＊＊ ]]> -20.54 Average width of crow's feet (mm) <![CDATA[-0.14 ＊＊＊ ]]> -18.68 Average depth of crow's feet wrinkles (mm) <![CDATA[-0.0053 ＊＊＊ ]]> -17.49

[0320] 2. Wrinkle improvement results at different time points after application of the test product

[0321] 2.1 Results of improvement in wrinkles around the eyes at different time points after use of the test product

[0322] After using the product for 1, 2, and 4 weeks, wrinkles in a fixed selected area around the corners of the eyes were analyzed. The results showed that the wrinkles around the corners of the eyes in the selected area were significantly improved before and after product use. See the results below:

[0323] Table 4.4 Results of the length, average width, and average depth of crow's feet at different time points after use of the test substance;

[0324] Table 4.5 Results of differences in crow's feet at different time points after use of the test substance;

[0325] as well as:

[0326] Figure 5 Bar chart showing the change in average length of crow's feet after use of the test product;

[0327] Figure 6Bar chart showing the change in average width of crow's feet after use of the test product;

[0328] Figure 7 Bar chart showing the change in average depth of crow's feet after use of the test substance.

[0329] Table 4.4 Results of length, average width, and average depth of crow's feet at different time points after application of the test substance.

[0330]

[0331]

[0332] Table 4.5 Results of differences in crow's feet at different time points after application of the test substance

[0333]

[0334] 2.2 Results of improvement in nasolabial folds at different time points after application of the test product

[0335] After using the product for 1, 2, and 4 weeks, wrinkles in a fixed area of ​​the nasolabial folds on the subjects' faces were analyzed. The results showed that the nasolabial folds in the selected area were significantly improved before and after product use compared to before use. The results are shown in the table below:

[0336] Table 4.6 Results of average width and average depth of nasolabial folds after use of the test material;

[0337] Table 4.7 Results of nasolabial fold differences at different time points after use of the test substance;

[0338] as well as:

[0339] Figure 8 Bar chart showing the change in average width of nasolabial folds after use of the test substance;

[0340] Figure 9 A bar chart showing the change in the average depth of nasolabial folds after the use of the test substance.

[0341] Table 4.6 Results of average width and average depth of nasolabial folds after use of the test material

[0342]

[0343]

[0344] Table 4.7 Results of nasolabial fold differences at different time points after use of the test substance

[0345]

[0346] 2.3 Results of improvement in forehead wrinkles at different time points after use of the test product

[0347] After using the product for 1, 2, and 4 weeks, wrinkles in a fixed area on the forehead of the subjects were analyzed. The results showed that the forehead wrinkles in the selected area were significantly improved before and after product use compared to before use. The results are shown below:

[0348] Table 4.6 Results of average width and average depth of nasolabial folds after use of the test material;

[0349] Table 4.7 Results of nasolabial fold differences at different time points after use of the test substance;

[0350] as well as:

[0351] Figure 10 Bar chart showing the change in average forehead wrinkle width after use of the test product;

[0352] Figure 11 Bar chart showing the change in average forehead wrinkle depth after use of the test product;

[0353] Figure 12 A bar chart showing the change in the average depth of forehead wrinkles after use of the test substance.

[0354] Table 4.8 Results of average width and average depth of forehead wrinkles after use of the test substance

[0355]

[0356]

[0357] Table 4.9 Results of forehead wrinkle differences at different time points after use of the test substance

[0358]

[0359] 3. Elasticity and firmness results at different time points after use of the test substance

[0360] 3.1 Results of elastic parameter R2

[0361] After 1, 2, and 4 weeks of product use, the skin elasticity of the subjects' cheeks was analyzed. The results showed that the facial elasticity parameter R2 of the subjects increased significantly compared with before use. The results are as follows:

[0362] Table 4.10 Results of elastic parameter R2 at different time points after use of the test material;

[0363] Table 4.11 Results of the difference in elastic parameter R2 at different time points after use of the test material;

[0364] as well as:

[0365] Figure 13 Bar graph showing the change in skin elasticity parameter R2 after use of the test substance.

[0366] Table 4.10 Results of elastic parameters R2 at different time points after use of the test material

[0367] parameter base value Week 1 Week 2 Week 4 Mean ± SD 0.46±0.096 0.47±0.073 0.51±0.074 0.54±0.052 Maximum value 0.67 0.61 0.70 0.69 Minimum value 0.29 0.36 0.41 0.46 median 0.45 0.48 0.51 0.55 p-value for normality test ＞0.05 ＞0.05 <0.05 ＞0.05 significance p-value / ＞0.05 <0.05 <0.001 Significance / ns * ***

[0368] Table 4.11 Results of the difference in elastic parameter R2 at different time points after use of the test material

[0369]

[0370] 3.2 Tightening parameter F4 results

[0371] After 1, 2, and 4 weeks of product use, the skin firmness of the subjects' cheeks was analyzed. The results showed that the subjects' facial firmness parameter F4 was significantly reduced compared with before use. The results are shown in Tables 11 and 12. Figure 13 The results are as follows:

[0372] Table 4.12 Results of the tightening parameter F4 at different time points after use of the test substance;

[0373] Table 4.13 Results of the difference in the tightening parameter F4 at different time points after use of the test substance;

[0374] as well as:

[0375] Figure 14 Bar chart showing the change in skin elasticity parameter F4 after use of the test substance.

[0376] Table 4.12 Results of F4 Tightening Parameters at Different Time Points After Use of the Test Substance

[0377] parameter base value Week 1 Week 2 Week 4 Mean ± SD 2.48±0.48 2.31±0.47 2.06±0.38 1.89±0.42 Maximum value 3.42 3.49 2.88 2.56 Minimum value 1.62 1.50 1.52 1.18 median 2.45 2.15 1.91 1.82 p-value for normality test ＞0.05 ＜0.05 ＜0.05 ＞0.05 significance p-value / ＞0.05 ＜0.001 ＜0.001 Significance / ns *** ***

[0378] Table 4.13 Results of the difference in the firming parameter F4 at different time points after use of the test substance

[0379]

[0380] 3.3 Gloss results of the test material at different time points after use

[0381] After using the product for 1, 2, and 4 weeks, the luster of the subjects' cheeks was analyzed. The results showed that the subjects' facial luster increased significantly compared to before use. See the results below:

[0382] Table 4.14 Gloss results of the test material at different time points after use;

[0383] Table 4.15 Results of gloss difference at different time points after use of the test material;

[0384] as well as:

[0385] Figure 15 Bar chart showing changes in skin glossiness after use of the test substance.

[0386] Table 4.14 Gloss results of the test material at different time points after use

[0387]

[0388]

[0389] Table 4.15 Results of gloss difference at different time points after use of the test material

[0390]

[0391] 4. Results of skin moisture content at different time points after application of the test substance

[0392] After 1, 2, and 4 weeks of product use, the skin moisture content of the subjects' cheeks was analyzed. The results showed that the moisture content of the stratum corneum on the subjects' faces was significantly increased compared with before use. The results are shown below:

[0393] Table 4.16 Results of skin stratum corneum moisture content at different time points after application of the test substance;

[0394] Table 4.17 Results of skin stratum corneum moisture difference at different time points after application of the test substance;

[0395] as well as:

[0396] Figure 16 Bar chart showing the water content of the stratum corneum of the skin.

[0397] Table 4.16 Results of skin stratum corneum moisture content at different time points after application of the test substance

[0398] parameter base value Week 1 Week 2 Week 4 Mean ± SD 52.37±10.53 56.99±9.68 64.64±8.00 69.96±6.13 Maximum value 68.80 70.15 82.57 81.92 Minimum value 24.25 36.73 45.20 59.92 median 52.58 58.85 64.73 69.33 p-value for normality test ＞0.05 ＜0.05 ＞0.05 ＞0.05 significance p-value / <0.01 ＜0.001 ＜0.001 Significance / ** *** ***

[0399] Table 4.17 Results of skin stratum corneum moisture difference at different time points after application of the test substance

[0400]

[0401] 5. After 1, 2, and 4 weeks of product use, the transepidermal water loss rate (TEWL) of the subjects' cheeks was analyzed. The results showed that the subjects' facial TEWL value (g / m2 / h) was significantly lower than before use. See the results below:

[0402] Table 4.18 Results of transepidermal water loss (TEWL) values ​​at different time points after use of the test substance;

[0403] Table 4.19 Results of the TEWL (transdermal water loss) difference at different time points after use of the test substance;

[0404] as well as:

[0405] Figure 17 Bar chart showing the changes in transepidermal water loss rate of the skin at different time points after the application of the test substance.

[0406] Table 4.18 Results of transepidermal water loss (TEWL) values ​​at different time points after application of the test substance

[0407] parameter base value Week 1 Week 2 Week 4 Mean ± SD 13.37±3.93 13.28±3.65 12.17±2.55 11.15±2.58 Maximum value 20.63 21.35 17.05 16.85 Minimum value 8.28 7.93 7.33 6.50 median 12.50 12.68 12.06 10.65 p-value for normality test ＜0.05 ＞0.05 ＞0.05 ＞0.05 significance p-value / ＞0.05 ＜0.05 ＜0.001 Significance / ns * ***

[0408] Table 4.19 Results of the difference in transdermal water loss (TEWL) at different time points after use of the test substance

[0409]

[0410] 6. Results of skin hemoglobin at different time points after application of the test substance

[0411] After 1, 2, and 4 weeks of product use, the skin moisture on the cheeks of the subjects was analyzed. The results showed that the hemoglobin levels in the facial skin of the subjects were significantly reduced compared with before use. The results are shown below:

[0412] Table 4.20 Results of skin hemoglobin at different time points after application of the test substance;

[0413] Table 4.21 Results of skin hemoglobin differences at different time points after application of the test substance;

[0414] as well as:

[0415] Figure 18 Bar chart showing changes in skin hemoglobin after use of the test substance.

[0416] Table 4.20 Results of skin hemoglobin at different time points after application of the test substance

[0417] parameter base value Week 1 Week 2 Week 4 Mean ± SD 307.00±37.35 298.18±44.42 283.66±46.39 276.06±41.87 Maximum value 371.00 377.00 371.33 359.00 Minimum value 240.00 227.00 183.17 181.33 median 308.17 286.19 281.33 271.83 p-value for normality test ＞0.05 ＞0.05 ＞0.05 ＞0.05 significance p-value / ＞0.05 ＜0.001 ＜0.001 Significance / ns *** ***

[0418] Table 4.21 Results of skin hemoglobin differences at different time points after application of the test substance

[0419]

[0420]

[0421] 7. Results of a* values ​​at different time points after use of the test substance

[0422] After 1, 2, and 4 weeks of product use, the skin moisture content of the subjects' cheeks was analyzed. The results showed that the a* value of the subjects' facial skin was significantly lower than before use. The results are as follows:

[0423] Table 4.22 Results of a* values ​​at different time points after use of the test substance;

[0424] Table 4.23 Results of the difference in a* values ​​at different time points after use of the test substance;

[0425] as well as:

[0426] Figure 19 Bar chart showing changes in skin a* values ​​after use of the test substance.

[0427] Table 4.22 Results of a* values ​​at different time points after use of the test substance

[0428] parameter base value Week 1 Week 2 Week 4 Mean ± SD 12.06±2.04 11.82±1.87 10.97±1.67 10.38±1.70 Maximum value 15.25 16.75 14.50 14.50 Minimum value 7.50 7.00 7.75 6.00 median 11.38 12.00 10.88 10.25 p-value for normality test ＞0.05 ＞0.05 ＞0.05 ＞0.05 significance p-value / ＞0.05 ＜0.01 ＜0.001 Significance / ns ** ***

[0429] Table 4.23 Results of a* value differences at different time points after use of the test substance

[0430]

[0431] V. Results of the Consumer Testing Questionnaire for Subjects

[0432] After the subjects used the product, they evaluated its effects on "suitability for sensitive skin", moisturizing, soothing, firming, and anti-wrinkle through a consumer use questionnaire. The following is the content of the questionnaire.

[0433] 1. The product is gentle (non-irritating) and suitable for sensitive skin.

[0434] During product use, none of the subjects experienced any adverse reactions or discomfort. The results are shown in Table 4.24, which presents the subjects' survey results regarding the product's suitability for sensitive skin.

[0435] Table 4.24 Results of the survey on the product's suitability for sensitive skin

[0436] serial number question agree p-value 1 After using the product, there was no discomfort on the face; the product is gentle and non-irritating. 100％ ＜0.001 2 This product is suitable for your sensitive skin. 100％ ＜0.001

[0437] 2. Skin feel after using the product

[0438] After the participants finished using the product, they were asked to fill in their overall feelings about the product and their skin satisfaction after using the product. The results are shown in Table 4.25, which is the survey results on the skin feel after using the product.

[0439] Table 4.25 Results of the survey on skin feel after using the product

[0440] serial number question agree p-value 1 The product has a refreshing and non-sticky texture. 100％ ＜0.001 2 The product is easily absorbed after use. 100％ ＜0.001

[0441] 3. Evaluation of product's moisturizing and gloss-enhancing effects

[0442] A consumer questionnaire survey was conducted on the subjects after they used the product for 4 weeks. Subjects were asked to evaluate the product’s moisturizing and glossing effects based on their experience using the product. The results are shown in Table 4.26, which shows the subjects’ survey results on the product’s claims on moisturizing and glossing effects.

[0443] Table 4.26 Survey Results on the Product's Claims Regarding Moisturizing and Glossiness

[0444]

[0445] 4. Evaluation of the product's anti-wrinkle efficacy

[0446] A consumer questionnaire survey was conducted on the subjects after they used the product for 4 weeks. Subjects were asked to evaluate the anti-wrinkle effect of the product on improving fine lines and wrinkles based on their experience using the product. The results are shown in Table 4.27.

[0447] Table 4.27 Survey Results on the Anti-wrinkle Efficacy Claims of the Products

[0448]

[0449]

[0450] 5. Product Firming Efficacy Evaluation

[0451] A consumer questionnaire survey was conducted on the subjects after using the product for 4 weeks. Subjects were asked to evaluate the product's skin-tightening efficacy based on their experience using the product. The results are shown in Table 4.28.

[0452] Table 4.28 Survey Results on Product Firming Efficacy Claims by Subjects

[0453]

[0454] 6. Evaluation of the product's soothing effects

[0455] A consumer questionnaire survey was conducted on the subjects after they used the product for 4 weeks. Subjects were asked to evaluate the product’s soothing effects based on their experience using the product. The results are shown in Table 4.29. Subjects’ survey results on the product’s claims of soothing and firming effects.

[0456] Table 4.29 Survey Results on the Product's Claims of Soothing and Firming Efficacy

[0457]

[0458] VI. Results Analysis

[0459] from Figure 20 A collection of photos showing changes in local facial features before and after use by some subjects. Figure 21 A collection of photos showing the improvement in wrinkles before and after use by some participants. Figure 22 Based on the 4-hour wrinkle improvement photos of some participants and the above observations of the participants, the following evaluation results can be obtained:

[0460] By observing 30 subjects aged 27-60 years, who used the test sample continuously for 4 weeks under normal conditions, the potential for adverse skin reactions was evaluated. Subjects' self-reported results showed no discomfort during or after use. The test results indicate that the Magic RNA Youthful Anti-Wrinkle Essence has a high safety profile, and it can be concluded that the anti-wrinkle essence provided in this embodiment is "mild (non-irritating)."

[0461] Four hours after using the product, the length of crow's feet decreased by 20.54% (p<0.05) compared to before use, which was statistically significant. The average width of crow's feet decreased by 18.68% (p<0.05), which was statistically significant, and the average depth of crow's feet decreased by 17.49% (p<0.05), which was statistically significant.

[0462] After one week of product use, the length of crow's feet decreased by 13.33% (p<0.05) compared to before use, showing a significant difference. The average width of crow's feet decreased by 17.87% (p<0.05), showing a significant difference compared to before use. The average depth of crow's feet decreased by 14.58% (p<0.05), showing a significant difference compared to before use. The average width of nasolabial folds decreased by 10.51% (p<0.05), showing a significant difference compared to before use; the average depth of nasolabial folds decreased by 11.97% (p<0.05), showing a significant difference compared to before use. The average width of forehead wrinkles decreased by 13.76% (p<0.05), showing a significant difference compared to before use; the average depth of forehead wrinkles decreased by 14.19% (p<0.05), showing a significant difference compared to before use.

[0463] After two weeks of product use, compared with before use, the length of crow's feet decreased by 17.55% (p<0.05), a significant difference; the average width of crow's feet decreased by 15.72% (p<0.05), a significant difference; the average depth of crow's feet decreased by 16.17% (p<0.05), a significant difference; the average width of nasolabial folds decreased by 13.03% (p<0.05), a significant difference; the average depth of nasolabial folds decreased significantly by 17.16% (p<0.05), a significant difference; the average width of forehead wrinkles decreased by 17.66% (p<0.05), a significant difference; and the average depth of forehead wrinkles decreased by 15.64% (p<0.05), a significant difference.

[0464] After 4 weeks of product use, the length of crow's feet decreased by 25.61% (p<0.05) compared to before use, showing a significant difference. The average width of crow's feet decreased by 17.71% (p<0.05), showing a significant difference compared to before use. The average depth of crow's feet decreased by 21.72% (p<0.05), showing a significant difference compared to before use. The average width of nasolabial folds decreased by 18.51% (p<0.05), showing a significant difference compared to before use. The average depth of nasolabial folds decreased by 21.98% (p<0.05), showing a significant difference compared to before use. The average width of forehead wrinkles decreased by 21.89% (p<0.05), showing a significant difference compared to before use. The average depth of forehead wrinkles decreased by 26.75% (p<0.05), showing a significant difference compared to before use. These results indicate that the product has anti-wrinkle effects.

[0465] After one week of product use, the subjects' transepidermal water loss (TEWL) value decreased by 0.64%, skin hemoglobin decreased by 2.87%, a* value decreased by 2.00%, and the stratum corneum moisture content increased by 8.84% (p<0.05), which were significantly different from before use. Glossiness increased by 3.54%.

[0466] After two weeks of product use, the subjects' transepidermal water loss (TEWL) value decreased by 8.96% (p<0.05), which was significantly different from before use; skin hemoglobin decreased by 4.73% (p<0.05), which was significantly different from before use; a* value decreased by 9.02% (p<0.05), which was significantly different from before use; skin stratum corneum moisture content increased by 14.61% (p<0.05), which was significantly different from before use; and skin gloss increased by 6.22% (p<0.05), which was significantly different from before use.

[0467] After 4 weeks of product use, the subjects experienced a significant decrease in transepidermal water loss (TEWL) by 16.59% (p<0.05) compared to pre-use levels, a significant decrease in skin hemoglobin by 10.08% (p<0.05), a significant decrease in a* value by 13.89% (p<0.05), a significant increase in stratum corneum hydration by 27.21% (p<0.05), and a significant increase in skin radiance by 15.10% (p<0.05). These results indicate that the product improves skin radiance and possesses moisturizing and repairing effects.

[0468] After one week of product use, the subjects' skin firmness F4 value decreased by 6.94%, and their elasticity index R2 value increased by 3.45%.

[0469] After two weeks of product use, the subjects' skin firmness F4 value decreased by 18.37% (p<0.05), which was significantly different from before use, and the elasticity index R2 value increased by 10.66% (p<0.05).

[0470] After 4 weeks of product use, the subjects' skin firmness F4 value decreased by 28.82% (p<0.05), which was significantly different from before use. The elasticity index R2 value increased by 17.47% (p<0.05). These results indicate that the product has a firming effect.

[0471] VII. Adverse Skin Reactions

[0472] During the use of the test substance, none of the 30 subjects experienced adverse skin reactions, and no subject withdrew from the trial due to failure to attend follow-up visits on time for personal reasons.

[0473] It should be noted that:

[0474] Although Examples 2, 3, and 4 only conducted further product safety evaluations on the anti-wrinkle serum of batch 240124F61 through human skin patch testing, and evaluated its effects on sensitive skin suitability, moisturizing, firming, anti-wrinkle, repairing, and radiance through two questionnaire tests, in fact, the same test evaluations can be performed on other batches of anti-wrinkle serums obtained according to the technical solution provided in the above embodiments, and the same safety evaluation and functional effect evaluation conclusions can be drawn. For the sake of simplicity, this instruction manual will not list more test data.

[0475] In summary, it can be seen that:

[0476] The preparation method provided by this invention can prepare an anti-wrinkle essence comprising polydeoxyribonucleic acid and human collagen as anti-wrinkle agents, ribonucleic acid as a neurotransmitter inhibitor, and antioxidants, moisturizers, thickeners, diluents, pH adjusters, and preservatives that can be accepted as cosmetic ingredients. Tests have shown that the anti-wrinkle essence prepared by the method provided by this invention not only meets the requirements of the "Cosmetic Safety Technical Specifications" (2015 edition) in all its physicochemical indicators, but also exhibits no adverse reactions after use. Therefore, it is safe, gentle, and non-irritating, suitable for sensitive skin. After use, it feels refreshing and non-sticky, easily absorbed, and not dry or tight. The skin is more moisturized, stronger, more radiant, softer, and more translucent, with significant moisturizing effects. The skin becomes more delicate, smooth, plump, and has the effect of improving rough skin, reducing facial fine lines and wrinkles, and soothing the skin.

[0477] Because the mechanism of wrinkle formation in human skin is clearly understood, the preparation method provided by this invention has a simple and clear formulation with appropriate and reasonable compatibility. In particular, the organic and ingenious combination of sodium DNA and human collagen as anti-wrinkle agents, and RNA as a neurotransmitter inhibitor, coupled with suitable production equipment and processes, can produce a gentle, soothing, and highly effective anti-wrinkle essence. It is not only safe and suitable for sensitive skin, but also has obvious moisturizing, anti-wrinkle, and skin-firming effects, as well as skin-soothing effects. Moreover, it is simple to prepare and easy to use, enabling the skin to quickly reduce wrinkles and maintain smoothness for a long time, allowing users to regain a youthful and beautiful appearance. Compared with existing technologies, it has outstanding beneficial effects and significant progress, and therefore has great promotion and application value.

[0478] The preparation method provided by this invention plays a crucial role in the preparation of anti-wrinkle serum because:

[0479] The anti-wrinkle serum prepared by this invention contains sodium DNase, human collagen, and RNA as a neurotransmitter inhibitor as its key anti-wrinkle active ingredients. However, 100 mL of the anti-wrinkle serum contains only 0.50–0.52 g of polydeoxyribonucleotides, only 0.10–0.12 g of human collagen, and even less 0.010–0.012 g of ribonucleic acid. Therefore, ensuring that these components are evenly dispersed and do not become damaged or lose their activity during the production process is not only related to the functional effect of the serum but also to the safety of the product.

[0480] The preparation method provided by this invention first involves adding carbomer as a thickener to a diluent. Then, sodium hyaluronate, glycerin, and butylene glycol as humectants, along with sclerotium tumefaciens as a thickener, are pre-dispersed and added to a mixing pot. The mixture is stirred and heated. Then, pentylene glycol as a humectant and an antioxidant are added. The mixture is stirred until the materials in the mixing pot are uniform. The temperature is then lowered for the first time, and the pH is adjusted. The temperature is then lowered for the second time to obtain a base liquid with a certain consistency and suitable temperature and pH. Finally, preservatives, neurotransmitter inhibitors, anti-wrinkle agents, and other materials as thickeners and humectants are added. This ensures that the anti-wrinkle agents and neurotransmitter inhibitors are not destroyed by high temperatures and can be slowly dispersed in the thicker base liquid, preventing them from quickly settling or adhering to the stirrer or the walls of the mixing pot, which would result in poor dispersion.

[0481] In the optimized preparation method, the preparation pot is first boiled in water for 30 minutes, then covered and cooled to room temperature before use. This ensures the hygiene and safety of the essence. Because the entire preparation process of the anti-wrinkle essence can only be carried out at a low temperature to ensure the performance of the materials, it is not possible to sterilize the materials by high temperature during the preparation process. Therefore, the pre-high temperature sterilization of the equipment is particularly important. Although there are various methods of sterilization in the existing technology, boiling in water will not damage the production equipment and will not leave any chemical residues. It is the most suitable pre-sterilization method for the anti-wrinkle essence to be prepared by this invention.

[0482] The stirring speed is set to 5-50 r / min, and a frame stirrer with a screen stirring frame is used in the optimized preparation method. This not only meets the stirring requirements of each stage of the preparation process, allowing the active ingredients to be fully dispersed, but also effectively removes particulate matter and trapped air bubbles in the preparation process of the essence.

[0483] Similarly, filtering through a sieve in the final stage of the preparation method can not only further remove particulate matter, entrained bubbles and foam from the essence, but also further disperse the various components in the essence, thereby obtaining a homogeneous and excellent product.

[0484] Based on the above explanation, it can be seen that:

[0485] The preparation method provided by this invention is adapted to the production of anti-wrinkle serums with multiple functions, especially those using sodium DNA, human collagen, and neurotransmitter inhibitor RNA as the main anti-wrinkle active ingredients. Furthermore, the preparation method of the anti-wrinkle serum provided by this invention is significantly different from the prior art, enabling the production of better anti-wrinkle serum products. The method is simple, the effect is excellent, and it represents a significant improvement over the prior art, making it highly valuable for promotion and application.

[0486] In the description process of the above instruction manual:

[0487] The terms “this embodiment,” “this case,” “as shown,” “further,” etc., are used to indicate that the specific features, structures, materials, or characteristics described in the embodiment or case are included in at least one embodiment or related case of the present invention.

[0488] In this specification, the illustrative expressions of the above terms are not necessarily directed at the same embodiments or cases, and the specific features, structures, materials or characteristics described may be combined or combined in any suitable manner in one or more embodiments or cases;

[0489] Furthermore, without creating contradictions, those skilled in the art can combine or integrate the different embodiments or cases described in this specification, as well as the features of the different embodiments or cases.

[0490] Finally, it should be noted that:

[0491] The above embodiments and effect examples are only used to illustrate the technical solutions of the present invention, and are not intended to limit them. Although the present invention has been described in detail with reference to the foregoing embodiments and effect examples, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments and effect examples, or equivalent substitutions can be made to some or all of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention. Non-essential improvements, adjustments or substitutions made by those skilled in the art based on the content of this specification are all within the scope of protection claimed by the present invention.

Claims

1. A method for preparing an anti-wrinkle serum, characterized in that: The anti-wrinkle serum comprises a clear and transparent mixture consisting of polydeoxyribonucleotides and human collagen as anti-wrinkle agents, ribonucleic acid as a neurotransmitter inhibitor, and antioxidants, moisturizers, thickeners, diluents, pH adjusters, and preservatives acceptable for addition to cosmetics. Its preparation method includes the following steps: Place the diluent in the mixing pot and slowly sprinkle in carbomer as a thickener. Sodium hyaluronate, glycerin, and butylene glycol, which are used as humectants, are pre-dispersed with sclerotium gum, which is used as a thickener, to prepare a uniformly dispersed mixture. Then, the mixture is added to the preparation pot, stirred and heated, and then pentylene glycol as a humectant and an antioxidant are added. Stirring continues until the material in the preparation pot is uniform, and then the first cooling is carried out. Add a pH adjuster to the prepared pot after the first cooling to adjust the pH, then perform a second cooling. Next, add preservatives, neurotransmitter inhibitors, anti-wrinkle agents, and other materials as thickeners and humectants. Continue stirring until homogeneous, let stand, and then filter through a sieve to obtain the final product.

2. The preparation method according to claim 1, characterized in that: The heating temperature is 70-75℃, the first cooling temperature is 50-55℃, and the second cooling temperature is below 40℃.

3. The preparation method according to claim 1, characterized in that: The stirring speed is 5-50 r / min, and the filtration screen has a mesh size of 200 mesh.

4. The preparation method according to claim 1, characterized in that: The preparation pot is a stainless steel steam-heated pot with a stirring device. The preparation pot is sterilized by boiling in water for 30 minutes, then sealed and cooled to room temperature before use.

5. The preparation method according to claim 4, characterized in that: The stirring device is a frame stirrer with a screen stirring frame.

6. The preparation method according to claim 1, characterized in that: The antioxidant is p-hydroxyacetophenone, the diluent is water, the pH adjuster is arginine, and the preservatives are glyceryl caprylate, capryloyl hydroxamic acid, and 1,3-propanediol.

7. The preparation method according to claim 6, characterized in that: 100mL of the anti-wrinkle essence contains 0.50-0.52g of the polydeoxyribonucleotide, 0.10-0.12g of the human collagen, 0.010-0.012g of the ribonucleic acid, 0.50-0.52g of the p-hydroxyacetophenone, 0.05-0.06g of the sodium hyaluronate, 6.0-6.2g of the glycerol, 4.0-4.2g of the butylene glycol, 3.0-3.2g of the pentylene glycol, 0.10-0.12g of the sclerotium tumefaciens gum, 0.10-0.12g of the carbomer, 0.10-0.12% of the arginine, and 0.30-0.32g of the combined amount of glyceryl caprylate, capryloyl hydroxamic acid, and 1,3-propanediol, with the remainder being water.

8. The preparation method according to claim 7, characterized in that: The anti-wrinkle essence is a clear, purple to purplish-red transparent liquid with a pH value of 6.5 to 7.5 and a relative density of 1.02 to 1.04 at 20°C.

9. The preparation method according to claim 7, characterized in that: According to Chapter 4 of the 2015 edition of the National Standard of the People's Republic of China "Technical Specifications for Safety of Cosmetics", the mercury content is ≤1mg / kg, the lead content is ≤10mg / kg, the arsenic content is ≤2mg / kg, and the cadmium content is ≤5mg / kg.

10. The preparation method according to claim 7, characterized in that: According to Chapter 5 of the 2015 edition of the National Standard of the People's Republic of China "Technical Specifications for Safety of Cosmetics", the total bacterial count should be ≤1000 CFU / g, the total mold and yeast count should be ≤100 CFU / g, and thermotolerant coliforms, Staphylococcus aureus and Pseudomonas aeruginosa should not be detected.

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