Composition containing collagen and sodium hyaluronate as well as preparation method and application of composition

Abstract

The invention discloses a composition containing collagen and sodium hyaluronate as well as a preparation method and application thereof, and the composition comprises the following components in percentage by weight: 0.1-2% of collagen, 1-3% of sodium hyaluronate, 0.05-0.1% of polydeoxyribonucleotide, 0.003-0.006% of sodium dihydrogen phosphate, 0.015-0.022% of disodium hydrogen phosphate, 0.6-0.95% of sodium chloride and water as a solvent. The collagen with the skin tightening effect, the sodium hyaluronate with the moisturizing effect and the polydeoxyribonucleotide with the collagen synthesis promoting effect are selected as effective components, and the effects of resisting aging, removing wrinkles and tightening and moisturizing the skin can be achieved.

Description

Technical Field

[0001] The present invention belongs to the technical field of skin care products, and in particular relates to a composition containing collagen and sodium hyaluronate, and a preparation method and application thereof. Background Art

[0002] Collagen, a biological polymer known scientifically as collagen, plays a role in connecting tissue in animal cells. It is one of the most critical raw materials in the biotechnology industry and a highly sought-after biomedical material. Its applications include biomedical materials, cosmetics, the food industry, and research. Collagen is primarily composed of three amino acids: glycine (Gly), proline (Pro), and hydroxyproline (Hyp). Collagen is closely linked to our skin's elasticity, hydration, wrinkles, and smoothness. Adults lose 1% of their collagen annually. Because women have less total collagen than men, this loss is more pronounced in women.

[0003] The molecular formula of sodium hyaluronate is (C 14 H 20 NO 11 Sodium hyaluronate (Na)n, composed of repeating disaccharides of D-glucuronic acid and N-acetyl-D-glucosamine, has widely varying molecular weights and swells in water to form a clear, viscous, elastic solution. Sodium hyaluronate behaves as a non-Newtonian fluid when dissolved in water, exhibiting excellent viscoelasticity, permeability, and biocompatibility. Sodium hyaluronate readily hydrolyzes to hyaluronic acid (HA) in water. Its natural abundance, degradability, and biocompatibility in animals and humans enable it to play a multifaceted role in regulating various biological processes and maintaining homeostasis, such as skin repair, cancer diagnosis, wound healing, tissue regeneration, anti-inflammatory properties, and immune regulation. Hyaluronic acid is highly hydrophilic and can capture approximately 1,000 times its weight in water. Its excellent hydration and moisture-locking properties make it a key ingredient in medical aesthetics and cosmetics.

[0004] Cosmetics are increasingly focusing on scientific formulations, maximizing product effectiveness through the combination of different ingredients. Currently, there are numerous functional products on the market, but most only have one or two benefits. Developing a product with multiple benefits would better meet market needs. Summary of the Invention

[0005] The present invention aims to provide a composition of collagen and sodium hyaluronate, its preparation method, and its use in skin care. The combined use of collagen and sodium hyaluronate has excellent ability to stimulate collagen regeneration and produces a synergistic enhancement effect. The addition of polydeoxyribonucleotides further enhances the composition's ability to stimulate collagen regeneration, achieving a firming and hydrating effect on the skin, thereby combining moisturizing, anti-wrinkle, and anti-aging effects. To achieve the above objectives, the present invention provides the following technical solutions: In one aspect, the present invention provides a composition of collagen and sodium hyaluronate, comprising the following components by weight percentage: 0.1-2% collagen, sodium hyaluronate, 1-3% polydeoxyribonucleotides, 0.05-1%, 0.003-0.006% sodium dihydrogen phosphate, 0.015-0.022% sodium hydrogen phosphate, 0.6-0.95% sodium chloride, and deionized water as a solvent.

[0006] In one or more embodiments of the present invention, the sodium hyaluronate includes a mixture of sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million.

[0007] Furthermore, in the compound, the mass ratio of sodium hyaluronate with a relative molecular mass of 0.3-10,000 to sodium hyaluronate with a relative molecular mass of 800,000-1.2 million is not higher than 4:6; further, the mass ratio is not higher than 3:7; further, the mass ratio is not higher than 2:8.

[0008] Furthermore, in the compound, the mass ratio of sodium hyaluronate with a relative molecular mass of 0.3-10,000 to sodium hyaluronate with a relative molecular mass of 800,000-1.2 million is 1:(7-9).

[0009] In the specific implementation process of the present invention, collagen is used. Collagen is closely related to the elasticity, hydration, wrinkles, smoothness and other aspects of the skin. Supplementing collagen can achieve the effect of firming the skin, anti-aging and wrinkle removal. In the specific implementation process of the present invention, polydeoxyribonucleotides are added to help promote the regeneration of collagen in skin tissue to quickly avoid skin sagging. On the other hand, the expression and synthesis of collagen are promoted through a synergistic mechanism, thereby improving the skin firming effect.

[0010] Furthermore, the collagen selected is type I human collagen or type III bovine collagen.

[0011] On the other hand, the present invention provides a method for preparing a composition of collagen and sodium hyaluronate, comprising: adding polydeoxyribonucleotides, a sodium hyaluronate composite composition, and collagen to a phosphate sodium chloride buffer solution in sequence, dissolving, filling, and sterilizing; the collagen content is 0.1-2%, the sodium hyaluronate content is 1-3%, and the polydeoxyribonucleotide content is 0.05-1%; the phosphate sodium chloride buffer solution contains sodium dihydrogen phosphate, disodium hydrogen phosphate, and sodium chloride.

[0012] The preparation method of the composition of collagen and sodium hyaluronate comprises the following steps: First, polydeoxyribonucleotides are dissolved in a phosphate sodium chloride buffer solution to form a transparent solution; sodium hyaluronate with a relative molecular mass of 0.3-10,000 and a relative molecular mass of 800,000-1.2 million are compounded and injected into the transparent solution containing polydeoxyribonucleotides to dissolve and form a transparent colloid; collagen is added and dissolved to form a milky white colloid; then a pH adjuster is added to adjust the pH to between 6.8 and 7.2 to obtain the final solution.

[0013] In a third aspect, the present invention provides an application of a composition containing collagen and sodium hyaluronate in the field of skin care. The composition containing collagen and sodium hyaluronate provided by the present invention is used as a liquid dressing in the application.

[0014] In the specific implementation process of the present invention, the preparation method includes the following steps: (1) First, dissolve the polydeoxyribonucleotide in phosphate-sodium chloride buffer solution to form a transparent solution; (2) Compound sodium hyaluronate with a mass ratio of 1:9, inject into phosphate sodium chloride buffer solution, and dissolve for 2-3 hours to form a transparent colloid; (3) Add collagen and dissolve it for 4-5 hours to form a milky white colloid; (4) Add a pH adjuster to adjust the pH value of the solution to between 6.8 and 7.2, and obtain a final solution with a pH value of 7.0.

[0015] In the specific implementation process of the present invention, the collagen selected is one of type I human collagen and type III bovine collagen. In the specific implementation process of the present invention, the pH regulator is selected from at least one of 1% sodium hydroxide solution, 0.1% sodium hydroxide solution, 0.1% hydrochloric acid solution and 1% hydrochloric acid solution. Compared with the prior art, the present invention has the following characteristics: (1) The present invention provides a composition of collagen and sodium hyaluronate and a preparation method thereof. The present invention adopts a compound of sodium hyaluronate, wherein low molecular weight sodium hyaluronate is conducive to better penetration of the skin, and medium and high molecular weight sodium hyaluronate is conducive to combining with skin tissue cells to form a good skin barrier and achieve better moisturizing effect. (2) The composition provided by the present invention, based on the synergistic effect of collagen and polydeoxyribonucleotides, exerts the best inhibitory effect on elastase, thereby achieving a firming effect. It combines moisturizing, anti-wrinkle and anti-aging effects.

[0016] (3) Comparative studies have found that the skin care composition provided by this application is composed of a variety of active ingredients with different functions in a certain proportion. The components work synergistically to achieve moisturizing, anti-wrinkle, and anti-aging effects. It can effectively increase skin moisturizing, enhance skin elasticity and skin barrier, improve skin firmness, improve rough and aging skin, effectively improve the instant anti-wrinkle effect of the skin, instantly smooth it, achieve anti-wrinkle and anti-aging, moisturizing and moisturizing effects, and make the skin more hydrated and plump. This product exerts its effects in all aspects and makes the effects in all aspects more obvious.

[0017] (4) The composition of collagen and sodium hyaluronate provided by the present invention is prepared from a mild and safe material, contains no chemical preservatives, and is non-irritating to the skin. DETAILED DESCRIPTION

[0018] The present invention is described in detail below with reference to the following embodiments. However, the following embodiments are only partial embodiments of the present invention, not all embodiments. Any limited modifications made within the scope of the claims of the present invention are still within the scope of the claims of the present invention. All other embodiments derived by persons of ordinary skill in the art based on the embodiments of the present invention are also within the scope of protection of the present invention.

[0019] Collagen was derived from type I human collagen. High molecular weight sodium hyaluronate in the sodium hyaluronate composition was purchased from Bloomage Biotechnology Co., Ltd. (J101230140), low molecular weight sodium hyaluronate was purchased from Bloomage Biotechnology Co., Ltd. (J201230424), and polydeoxyribonucleotide was purchased from Ruijiming Biotechnology Co., Ltd. (20240517).

[0020] Example 1, a composition containing collagen and sodium hyaluronate Calculated by weight percentage, it includes: 2% collagen, 1.5% sodium hyaluronate composition, 0.5% polydeoxyribonucleotide, 0.005% sodium dihydrogen phosphate, 0.022% disodium hydrogen phosphate, 0.9% sodium chloride, and the balance is deionized water.

[0021] The sodium hyaluronate composition described in this embodiment includes sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million, with a mass ratio of 1:9. The collagen described in this embodiment is type I collagen. The solvent in this embodiment is a phosphate sodium chloride buffer solution, wherein the mass ratio of sodium dihydrogen phosphate, sodium hydrogen phosphate, and sodium chloride is 0.05:0.22:9, and the concentrations are 0.005%, 0.022%, and 0.9%, respectively, and the balance is deionized water. First, the polydeoxyribonucleotide is dissolved in a phosphate sodium chloride buffer solution to form a transparent solution; sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million is compounded in a mass ratio of 1:9, and injected into the transparent solution containing polydeoxyribonucleotide, and dissolved for 2-3 hours to form a transparent colloid; type I collagen is added and dissolved for 4-5 hours to form a milky white colloid; then 1% sodium hydroxide solution is added to adjust the pH to 7.0 to obtain the final solution.

[0022] Example 2, a composition containing collagen and sodium hyaluronate Calculated by weight percentage, it includes: 1.5% collagen, 1.5% sodium hyaluronate composition, 0.5% polydeoxyribonucleotide, 0.005% sodium dihydrogen phosphate, 0.022% disodium hydrogen phosphate, 0.9% sodium chloride, and the balance is deionized water.

[0023] The sodium hyaluronate composition described in this embodiment includes sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million, with a mass ratio of 1:9. The collagen described in this embodiment is type I collagen. The solvent in this embodiment is a phosphate sodium chloride buffer solution, wherein the mass ratio of sodium dihydrogen phosphate, disodium hydrogen phosphate, and sodium chloride is 0.05:0.22:9, and the balance is deionized water.

[0024] The pH regulator in this embodiment is 1% sodium hydroxide solution. First, the polydeoxyribonucleotide is dissolved in a phosphate sodium chloride buffer solution to form a transparent solution; sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million is compounded in a mass ratio of 1:9, and injected into the transparent solution containing polydeoxyribonucleotide, and dissolved for 2-3 hours to form a transparent colloid; type I collagen is added and dissolved for 4-5 hours to form a milky white colloid; then 1% sodium hydroxide solution is added to adjust the pH to 7.0 to obtain the final solution. Example 3, a composition containing collagen and sodium hyaluronate Calculated by weight percentage, it includes: 0.1% collagen, 1.5% sodium hyaluronate composition, 0.5% polydeoxyribonucleotide, 0.005% sodium dihydrogen phosphate, 0.022% disodium hydrogen phosphate, 0.9% sodium chloride, and the balance is deionized water. The sodium hyaluronate composition described in this embodiment includes sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million, with a mass ratio of 1:9. The collagen described in this embodiment is type I collagen. The solvent in this embodiment is a phosphate sodium chloride buffer solution, wherein the mass ratio of sodium dihydrogen phosphate, disodium hydrogen phosphate, and sodium chloride is 0.05:0.22:9, and the balance is deionized water. The pH regulator in this embodiment is 1% sodium hydroxide solution.

[0025] First, the polydeoxyribonucleotide is dissolved in a phosphate sodium chloride buffer solution to form a transparent solution; sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million is compounded in a mass ratio of 1:9, and injected into the transparent solution containing polydeoxyribonucleotide, and dissolved for 2-3 hours to form a transparent colloid; type I collagen is added and dissolved for 4-5 hours to form a milky white colloid; then 1% sodium hydroxide solution is added to adjust the pH to 7.0 to obtain the final solution.

[0026] Example 4 In a first aspect of this embodiment, a composition containing collagen and sodium hyaluronate is provided, which comprises, by weight percentage, 2% collagen, 1% sodium hyaluronate composition, 0.5% polydeoxyribonucleotide, 0.005% sodium dihydrogen phosphate, 0.022% disodium hydrogen phosphate, 0.9% sodium chloride, and the balance being deionized water. The sodium hyaluronate composition described in this embodiment includes sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million, with a mass ratio of 1:9. The collagen described in this embodiment is type I collagen. The solvent in this embodiment is a phosphate sodium chloride buffer solution, wherein the mass ratio of sodium dihydrogen phosphate, disodium hydrogen phosphate, and sodium chloride is 0.05:0.22:9, and the balance is deionized water.

[0027] The pH regulator in this embodiment is 1% sodium hydroxide solution.

[0028] First, the polydeoxyribonucleotide is dissolved in a phosphate sodium chloride buffer solution to form a transparent solution; sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million is compounded in a mass ratio of 1:9, and injected into the transparent solution containing polydeoxyribonucleotide, and dissolved for 2-3 hours to form a transparent colloid; type I collagen is added and dissolved for 4-5 hours to form a milky white colloid; then 1% sodium hydroxide solution is added to adjust the pH to 7.0 to obtain the final solution. Example 5, a composition containing collagen and sodium hyaluronate Calculated by weight percentage, it includes: 2% collagen, 3% sodium hyaluronate composition, 0.5% polydeoxyribonucleotide, 0.005% sodium dihydrogen phosphate, 0.022% disodium hydrogen phosphate, 0.9% sodium chloride, and the balance is deionized water. The sodium hyaluronate composition described in this embodiment includes sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million, with a mass ratio of 1:9.

[0029] The collagen described in this embodiment is type I collagen.

[0030] The solvent in this embodiment is a phosphate sodium chloride buffer solution, wherein the mass ratio of sodium dihydrogen phosphate, disodium hydrogen phosphate, and sodium chloride is 0.05:0.22:9, and the balance is deionized water.

[0031] The pH regulator in this embodiment is 1% sodium hydroxide solution. First, the polydeoxyribonucleotide is dissolved in a phosphate sodium chloride buffer solution to form a transparent solution; sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million is compounded in a mass ratio of 1:9, and injected into the transparent solution containing polydeoxyribonucleotide, and dissolved for 2-3 hours to form a transparent colloid; type I collagen is added and dissolved for 4-5 hours to form a milky white colloid; then 1% sodium hydroxide solution is added to adjust the pH to 7.0 to obtain the final solution. Comparative Example 1 The specific operation of this comparative example is the same as that of the above embodiment, except that: the content of the sodium hyaluronate composition is 1.5% (sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million, with a mass ratio of 1:9), 0.5% polydeoxyribonucleotide, 0.005% sodium dihydrogen phosphate, 0.022% sodium hydrogen phosphate, 0.9% sodium chloride, and the balance is deionized water. Comparative Example 2 The specific operation of this comparative example is the same as that of the above embodiment, except that: collagen 2%, polydeoxyribonucleotide 0.5%, sodium dihydrogen phosphate 0.005%, sodium hydrogen phosphate 0.022%, sodium chloride 0.9%, and the balance is deionized water. Comparative Example 3 The specific operation of this comparative example is the same as that of the above embodiment, except that: 2% collagen, 1.5% sodium hyaluronate composition (sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million, compounded in a mass ratio of 1:9), 0.005% sodium dihydrogen phosphate, 0.022% disodium hydrogen phosphate, 0.9% sodium chloride, and the balance is deionized water. Comparative Example 4 The specific operation of this comparative example is the same as that of the above embodiment, except that: 1.5% sodium hyaluronate composition (sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million, with a mass ratio of 1:9), 0.005% sodium dihydrogen phosphate, 0.022% disodium hydrogen phosphate, 0.9% sodium chloride, and the balance is deionized water. Comparative Example 5 The specific operation of this comparative example is the same as that of the above embodiment, except that: 2% collagen, 1.5% sodium hyaluronate composition (a mixture of sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million, with a mass ratio of 1:7), 0.5% polydeoxyribonucleotide, 0.005% sodium dihydrogen phosphate, 0.022% sodium hydrogen phosphate, 0.9% sodium chloride, and the balance being deionized water. Comparative Example 6 The specific operation of this comparative example is the same as that of the above embodiment, except that: 2% collagen, 1.5% sodium hyaluronate composition (a mixture of sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million, with a mass ratio of 1:10), 0.5% polydeoxyribonucleotide, 0.005% sodium dihydrogen phosphate, 0.022% sodium hydrogen phosphate, 0.9% sodium chloride, and the balance being deionized water.

[0032] Comparative Example 7 The specific operation of this comparative example is the same as that of the above embodiment, except that: 2% collagen, 1.5% sodium hyaluronate composition (a mixture of sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million, with a mass ratio of 0:10), 0.5% polydeoxyribonucleotide, 0.005% sodium dihydrogen phosphate, 0.022% disodium hydrogen phosphate, 0.9% sodium chloride, and the balance is deionized water.

[0033] Comparative Example 8 The specific operation of this comparative example is the same as that of the above embodiment, except that: 2% collagen, 1% sodium hyaluronate composition (a mixture of sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million, with a mass ratio of 1:10), 0.5% polydeoxyribonucleotide, 0.005% sodium dihydrogen phosphate, 0.022% sodium hydrogen phosphate, 0.9% sodium chloride, and the balance being deionized water.

[0034] Comparative Example 9 The specific operation of this comparative example was the same as that of the above example, with the following differences: 2% collagen, 1% sodium hyaluronate composition (a mixture of sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million, with a mass ratio of 1:7), 0.5% polydeoxyribonucleotides, 0.005% sodium dihydrogen phosphate, 0.022% disodium hydrogen phosphate, 0.9% sodium chloride, and the balance being deionized water. The order of adding the polydeoxyribonucleotide and sodium hyaluronate composite composition was adjusted, with the sodium hyaluronate composite added first, followed by the polydeoxyribonucleotide. It was found that when the order of adding the polydeoxyribonucleotide and sodium hyaluronate was adjusted, precipitation was produced.

[0035] Comparative Example 10 The specific operation of this comparative example is the same as that of the above embodiment, except that: 2% collagen, 3% sodium hyaluronate composition (a mixture of sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million, with a mass ratio of 1:10), 0.5% polydeoxyribonucleotide, 0.005% sodium dihydrogen phosphate, 0.022% sodium hydrogen phosphate, 0.9% sodium chloride, and the balance being deionized water.

[0036] Comparative Example 11 The specific operation of this comparative example is the same as that of the above embodiment, except that: 2% collagen, 3% sodium hyaluronate composition (a mixture of sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million, with a mass ratio of 1:7), 0.5% polydeoxyribonucleotide, 0.005% sodium dihydrogen phosphate, 0.022% sodium hydrogen phosphate, 0.9% sodium chloride, and the balance being deionized water.

[0037] The material amounts of the embodiments and comparative examples are summarized in Table 1: Table 1, Component types and ratios (%) of Examples 1-5 and Comparative Examples 1-11

[0038] Test case, efficacy test 1. Elastase test Experimental background The firming effect is mainly characterized by evaluating the inhibition rate of the test sample on elastase. Elastase is mainly synthesized and secreted by fibroblasts, and can degrade elastin in the skin, causing skin aging. The experimental principle of inhibiting elastase is that porcine pancreatic elastase catalyzes the reaction with the enzyme substrate. After adding the active substance, the absorbance changes, and the magnitude of the absorbance change reflects the inhibition rate of the elastase inhibitor. This experiment compares the results of the elastase inhibition rate test of the test sample with the negative control. If the inhibition rate of the test sample is higher than that of the negative control and there is a significant difference (P < 0.05), the test sample can be considered to have a firming effect.

[0039] Scope of application and relevance This in vitro test is suitable for evaluating cosmetic products and ingredients claiming to achieve firming and anti-wrinkle effects through elastase inhibition. This in vitro method complements subjective and objective human evaluation methods, as it is the primary pathway for elastin degradation in the skin and correlates well with in vivo firming efficacy testing. Experimental setup Sample group test: dilute with pure water to the concentration required for sample testing; Negative control: pure water; Positive control: EGCG was diluted with water to a concentration of 0.1%. Experimental methods Set up sample group, sample background group, solvent group and solvent background group, and set up 3 parallel groups for each group. (1) Add 0.2 mL of sample solution of the same concentration to each of the sample group and the sample background group. Add 1.0 mL of pH 8.8 boric acid buffer to the sample background group, 0.2 mL of pH 8.8 boric acid buffer to the solvent group, and 1.0 mL of pH 8.8 boric acid buffer to the solvent background group. (2) Add 10 mL of AAPAN solution and 1.0 mL of elastase to each of the sample and solvent groups. Mix the mixture thoroughly and shake in a constant temperature water bath for 20 min.

[0040] (3) 2 mL of pH 6.0 phosphate buffer was added to each of the sample group, sample background group, solvent group, and solvent background group. Then, an equal amount of a mixture of pH 8.8 boric acid buffer and pH 6.0 phosphate buffer was added to make the volume 5 mL. The mixture was centrifuged for 5 min and 3 mL of the supernatant was collected. (4) Transfer each reaction solution into a 1 cm cuvette and measure the absorbance at 410 nm. Calculation formula

[0041] Where: A1 is the absorbance of the reaction solution without sample; A2 is the absorbance of the reaction solution without sample and enzyme; A3 is the absorbance of the reaction solution containing the sample and enzyme; A4 is the absorbance of the reaction solution containing the sample and without the enzyme. Data Analysis Statistical analysis was performed using SPSS software. Comparisons of elastase inhibition rates between the test samples, the positive control group, and the negative control group were performed using independent sample t-tests. All statistical analyses were performed using two-tailed tests, with a significance level of α = 0.05. A P > 0.05 indicated no significant difference between the two groups; a P < 0.05 indicated a significant difference between the two groups. The compositions obtained in Examples 1-5 and Comparative Examples 1-7 were tested according to the above test method. The results are shown in Table 2.

[0042] Table 2. Elastase test data and analysis of the compositions

[0043] According to the results in Table 2, the data of Examples 1-5 show that when the firming ingredients collagen and polydeoxyribonucleotides are present, the inhibitory effect on elastase is significant, and when the collagen content decreases, the elastase inhibition rate decreases. The elastase inhibition rate results of Examples 1-5 are significantly different from those of the negative control group (P<0.05), indicating that they have a firming effect.

[0044] The results of Comparative Example 1 show that when the collagen component is missing, the inhibition rate is less than 50%, and the inhibitory effect on elastin is not obvious. The results of Comparative Example 4 showed that when collagen and polydeoxyribonucleotides were absent, the inhibition rate on elastin was similar to that of the negative control, indicating that the two played a synergistic role.

[0045] The results of Comparative Example 9 show that when the order of adding polydeoxynucleotides and sodium hyaluronate is adjusted, precipitation occurs, resulting in a decrease in the effect of polydeoxynucleotides on promoting collagen production and a lower elastase inhibition rate.

[0046] 2. Human body firming efficacy test Test method: There are 12 experimental groups, recruiting a total of 25×12 volunteers; ensuring that the final recovery data meets the requirements of more than 20 people in each group.

[0047] Table 3, Test Equipment Information

[0048] Take an appropriate amount of sample and apply evenly on the face, gently massage until completely absorbed, apply once in the morning and evening. Test Evaluation Skin elasticity R2 value Parameter description: Reflects the ability of the skin to return to its original shape after being stretched or compressed; the larger the measured value, the better the skin elasticity. Result judgment: Before and after trial use of the product, the R2 value of skin elasticity increased significantly (P < 0.05), indicating that the test substance has the effect of improving skin elasticity.

[0049] Skin firmness F4 value: Using an elasticity tester, the logarithmic curve area of ​​the skin's overall amplitude during ten consecutive negative pressure cycles.

[0050] Parameter description: Reflects the firmness of the skin; the smaller the measured value, the firmer the skin.

[0051] Descriptive statistics were performed for each test value using SPSS software. The Shapiro-Wilk test was used to test for significance of normal distribution of data changes. If P > 0.05, the data were normally distributed. A paired sample t-test was performed with a significance level of α = 0.05. If P < 0.05, the data were non-normally distributed, and a Wilcoxon signed-rank test was performed. The test results are shown in Table 4; Table 4, Test Equipment Information

[0052] According to the results in Table 4, the data of Examples 1-5 show that when the firming ingredients collagen and polydeoxyribonucleotides are present, the effect of improving skin elasticity is significant, and when the collagen content decreases, the change in skin elasticity gradually decreases, indicating that collagen has the effect of improving skin elasticity. The results of Comparative Example 1 show that when the collagen component is lacking, the change rate is small, but due to the presence of polydeoxyribonucleotides, it shows a certain effect on improving skin elasticity, indicating that deoxyribonucleotides have a certain effect on improving skin elasticity.

[0053] The results of Comparative Example 4 show that when collagen and polydeoxyribonucleotides are lacking, the rate of change of skin elasticity is similar to that of the blank control, and the sample does not show the efficacy of improving skin elasticity, indicating that the combined use of the two has a synergistic effect.

[0054] The results of Comparative Example 9 show that when the order of adding polydeoxynucleotides and sodium hyaluronate is adjusted, precipitation occurs, resulting in a decrease in the effect of polydeoxynucleotides on promoting collagen production and a decrease in the rate of change, indicating that it has a certain effect on improving skin elasticity. 3. Skin moisturizing efficacy test Testers: 120 female volunteers aged 38-55 years old with dry, dull and yellow skin were randomly divided into 12 groups, with 10 people in each group. Each subject was informed of the entire research process and signed an informed consent form. Test samples: skin care composition samples prepared in Examples 1-5 and Comparative Examples 1-7. Testing Method: After cleansing their faces once in the morning and evening, each group of 10 participants applied one sample to their faces. The skin care compositions prepared in Examples 1-5 and Comparative Examples 1-7 were applied evenly to their faces. No other cosmetics were used during the testing period, which lasted for four weeks. Facial skin moisture content and transepidermal water loss (TEWL) were measured before and after application, with measurements taken every two weeks and the average value calculated.

[0055] All test subjects cleaned their faces on days 0, 14, and 28, rested for 30 minutes in the test environment, and then underwent the following tests: (1) facial skin moisture content was measured using the MoistureMeterSC (Delfin, Finland) skin stratum corneum moisture meter, and the value displayed on the instrument was read; (2) transepidermal water loss (TEWL value) was measured using the VapoMeter (Delfin, Finland) skin moisture loss meter, and the value displayed on the instrument was read.

[0056] Test environment requirements: temperature 21±1℃; relative humidity 50±5%RH; lighting conditions: color temperature 5500-6500K. Test results: (1) Facial skin moisture content test The test results of facial skin moisture content are shown in Table 5. The change value after 28 days (%) = (facial skin moisture content on day 28 - facial skin moisture content on day 0) / facial skin moisture content on day 0 × 100%.

[0057] Table 5, facial skin moisture content test results of various skin care product compositions

[0058] Comparative Examples 6 to 8 have significant elastase inhibition effects, good firming effects, and low transepidermal water loss values, indicating that they have skin moisturizing effects. However, the skin water content is low. The reason is that the low content of low molecular weight HA leads to a reduced hydrating effect, and therefore, the skin water content is low.

[0059] (2) Transepidermal water loss (TEWL value) test The results of the transepidermal water loss (TEWL) test are shown in Table 6. The change after 28 days (%) = (TEWL value at 28 days - TEWL value at 0 days) / TEWL value at 0 days × 100. Table 6, Transepidermal Water Loss (TEWL) test results of various skin care compositions

[0060] As can be seen from Examples 1-5 in Tables 5 and 6, as the content of sodium hyaluronate increases, the moisture content of the facial skin gradually increases, and the transepidermal water loss value of the skin decreases, indicating that the skin moisturizing effect is good and the skin barrier function is enhanced. In Comparative Example 2, no sodium hyaluronate was added, and the skin water content was significantly reduced, and the transepidermal water loss value of the skin was reduced, indicating that the sample did not achieve the moisturizing effect and the skin barrier function was weakened. The results show that sodium hyaluronate has moisturizing effects.

[0061] In Comparative Example 7, only high molecular weight sodium hyaluronate was added. The results show that the hydrating effect was not obvious, but the moisturizing effect was relatively good in reducing water loss.

[0062] In Comparative Example 9, the order of adding polydeoxyribonucleotides and sodium hyaluronate was adjusted, resulting in a decrease in the effective amount of sodium hyaluronate used, and thus a weakened moisturizing effect.

[0063] The above description is only a preferred embodiment of the present application and is not intended to limit the scope of protection of the present application. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present application are included in the scope of protection of the present application.

Claims

1. A composition of collagen and sodium hyaluronate, characterized in that: The invention comprises the following components by weight percentage: 0.1-2% collagen, 1-3% sodium hyaluronate, 0.05-1% polydeoxyribonucleotide, 0.003-0.006% sodium dihydrogen phosphate, 0.015-0.022% disodium hydrogen phosphate, 0.6-0.95% sodium chloride, and deionized water as a solvent; the sodium hyaluronate comprises a compound of sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million.

2. The composition of collagen and sodium hyaluronate according to claim 1, characterized in that In the compound, the mass ratio of sodium hyaluronate with a relative molecular mass of 0.3-10,000 to sodium hyaluronate with a relative molecular mass of 800,000-1.2 million is not higher than 4:

6.

3. The composition of collagen and sodium hyaluronate according to claim 1, characterized in that In the compound, the mass ratio of sodium hyaluronate with a relative molecular mass of 0.3-10,000 to sodium hyaluronate with a relative molecular mass of 800,000-1.2 million is not higher than 3:7, and further, the mass ratio is not higher than 2:

8.

4. The composition of collagen and sodium hyaluronate according to claim 1, characterized in that In the compound, the mass ratio of sodium hyaluronate with a relative molecular mass of 0.3-10,000 to sodium hyaluronate with a relative molecular mass of 800,000-1.2 million is 1:7-9.

5. The composition of collagen and sodium hyaluronate according to claim 1, characterized in that The collagen used is type I human collagen or type III bovine collagen.

6. A method for preparing a composition of collagen and sodium hyaluronate, characterized in that: include: The preparation method is prepared by sequentially adding polydeoxyribonucleotide, sodium hyaluronate compound and collagen into a phosphate sodium chloride buffer solution, dissolving, filling and sterilizing; the collagen content is 0.1-2%, the sodium hyaluronate content is 1-3%, and the polydeoxyribonucleotide content is 0.05-1%; the phosphate sodium chloride buffer solution contains sodium dihydrogen phosphate, disodium hydrogen phosphate and sodium chloride.

7. The method for preparing the composition of collagen and sodium hyaluronate according to claim 6, wherein: The steps include: First, polydeoxyribonucleotides are dissolved in a phosphate sodium chloride buffer solution to form a transparent solution; sodium hyaluronate with a relative molecular mass of 0.3-10,000 and a relative molecular mass of 800,000-1.2 million are compounded and injected into the transparent solution containing polydeoxyribonucleotides to dissolve and form a transparent colloid; collagen is added and dissolved to form a milky white colloid; then a pH adjuster is added to adjust the pH to between 6.8 and 7.2 to obtain the final solution.

8. The method for preparing the composition of collagen and sodium hyaluronate according to claim 6, wherein: The composition of collagen and sodium hyaluronate comprises, by weight percentage, 2% collagen, 1.5% sodium hyaluronate composition, 0.5% polydeoxyribonucleotide, 0.005% sodium dihydrogen phosphate, 0.022% disodium hydrogen phosphate, 0.9% sodium chloride, and the balance being deionized water; First, the polydeoxyribonucleotide is dissolved in a phosphate sodium chloride buffer solution to form a transparent solution; sodium hyaluronate with a relative molecular mass of 0.3-10,000 and sodium hyaluronate with a relative molecular mass of 800,000-1.2 million is compounded in a mass ratio of 1:9, and injected into the transparent solution containing the polydeoxyribonucleotide and dissolved for 2-3 hours to form a transparent colloid; type I collagen is added and dissolved for 4-5 hours to form a milky white colloid; then the pH value is adjusted to 7.0 to obtain the final solution.

9. Use of the composition of collagen and sodium hyaluronate according to any one of claims 1 to 5 in the field of skin care.

10. The use according to claim 9, characterized in that The composition of collagen and sodium hyaluronate is used as a liquid dressing.

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