Traditional Chinese medicine protein peptide composition with whitening, anti-aging and moisturizing effects, preparation method and application of traditional Chinese medicine protein peptide composition in cosmetics

Abstract

The invention discloses a traditional Chinese medicine protein peptide composition with whitening, anti-aging and moisturizing effects, a preparation method and application of the traditional Chinese medicine protein peptide composition in cosmetics. The traditional Chinese medicine protein peptide composition is composed of rhizoma polygonati protein peptide and radix polygonati officinalis protein peptide which are obtained through an enzymolysis method and have the molecular weight of 10 KDa or below, and the two protein peptides have the synergistic interaction effect on the whitening, anti-aging and moisturizing effects. The invention also provides a preparation method of the polygonatum sibiricum protein peptide and the polygonatum odoratum protein peptide, a mild and environment-friendly production process is realized through a multi-time enzymolysis technology and an optimized preparation process, the biological activity of the traditional Chinese medicine protein peptide is maintained to the maximum extent, the purity of the protein peptide is improved, and no irritation is ensured. The traditional Chinese medicine protein peptide composition is applied to cosmetics of different dosage forms such as water aqua, emulsion, cream and essence, is wide in application, provides a multifunctional, non-irritant, natural and mild raw material selection for the cosmetic industry, and meets the urgent demand of consumers for safe, efficient and environment-friendly cosmetics.

Description

Technical Field

[0001] The present invention belongs to the technical field of cosmetics, and in particular, relates to a traditional Chinese medicine protein peptide composition with whitening, anti-aging and moisturizing effects, a preparation method and application thereof in cosmetics. Background Art

[0002] In the rapidly changing beauty and health market, consumers' demand for cosmetics has far exceeded basic cleansing and moisturizing, and they are pursuing safer, more efficient, multifunctional and environmentally friendly products. This trend has prompted the cosmetics industry to continuously explore new sources of raw materials and preparation technologies to meet consumers' growing personalized needs. Traditional Chinese medicine, a natural resource with rich bioactive ingredients and a long history of application, is gradually becoming a new hot spot in cosmetics research and development.

[0003] According to the National Food Safety Standard Plant Protein Peptides for Food Processing (GB 31611-2023), Chinese medicine protein peptides are products with peptides with a relative molecular mass of 189 to 10,000 as the main component, which are produced from protein-rich Chinese medicinal materials through extraction, enzymatic hydrolysis and (or) microbial fermentation, filtration, sterilization, drying and other processes. These peptides are usually composed of 2 to 50 amino acid residues, with a small molecular weight and rich variety. They have the characteristics of easy cell transport, high absorption and utilization, short half-life, low toxicity and immunogenicity, and diverse biological activities. Therefore, they show excellent application potential in the fields of food and cosmetics. For example, protein peptides extracted from traditional Chinese medicines such as ginseng, astragalus, and yam can be used to develop products that improve human resistance because of their ability to regulate the immune system. Protein peptides extracted from Chinese wolfberry, hawthorn and other medicinal materials can not only provide additional nutritional supplements, but also improve the taste and flavor of food; protein peptides extracted from Ganoderma lucidum, ginseng, Schisandra chinensis and other medicinal materials have antioxidant activity, which can effectively resist the damage of free radicals to the skin and delay the aging process of the skin. In short, Chinese medicine protein peptides are extracted and enzymatically hydrolyzed from Chinese medicinal materials through modern biotechnology. Because of their small molecular weight and easy absorption, they have biological activities such as immunomodulation, anti-oxidation, and elimination of free radicals, and show great application potential in the fields of food and cosmetics.

[0004] Polygonatum sibiricum is the dried rhizome of Polygonatum kingianum Coll.et Hemsl., Polygonatum sibiricum Red. or Polygonatum cyrtonema Hua, all of which are plants of the Liliaceae family. Polygonatum odoratum is the dried rhizome of Polygonatum odoratum (Mill.) Druce, all of which are plants of the Liliaceae family. Polygonatum sibiricum and Polygonatum odoratum are traditional Chinese medicinal materials, and they have the effects of nourishing yin and moistening the lungs, nourishing the skin and beautifying the skin in traditional Chinese medicine theory. Modern studies have shown that both Polygonatum sibiricum and Polygonatum odoratum contain rich active ingredients such as polysaccharides, proteins, and amino acids, which play an important role in improving skin conditions and enhancing skin immunity. In particular, Polygonatum sibiricum protein peptides and Polygonatum odoratum protein peptides extracted and hydrolyzed by modern biotechnology have their molecular weights reduced by enzymatic hydrolysis technology, thereby improving skin permeability and bioavailability.

[0005] Both Polygonatum sibiricum and Polygonatum odoratum contain a lot of protein and polysaccharide components, and the two are mostly combined into glycoproteins. Glycoproteins are biological macromolecules composed of sugars and proteins. To obtain protein peptides from Polygonatum sibiricum and Polygonatum odoratum, it is necessary to decompose the glycoproteins into sugars and proteins, remove the sugars, and further enzymatically hydrolyze the resulting proteins. At present, there are few studies on the extraction and preparation processes of Polygonatum sibiricum and Polygonatum odoratum protein peptides, and there are certain problems. For example, the preparation process uses extreme conditions such as high temperature, high pressure, strong acid and strong alkali chemical hydrolysis, and power supply, which cannot guarantee the activity of the protein peptides; or the crude protein is not extracted in advance, but the Chinese medicine liquid is directly enzymatically hydrolyzed and filtered, and the final product contains a large number of other components with a molecular weight close to that of the protein peptide, and the purity of the obtained protein peptide is low.

[0006] CN 110934837A discloses a composite tablet of Polygonatum sibiricum polypeptide and a preparation method thereof. The preparation method of Polygonatum sibiricum polypeptide is to add Polygonatum sibiricum extract to an expansion solvent composed of CO2 and ethanol, pressurize to 1.5-2.5MPa to form an expansion system for high-pressure extraction. CN106083990A discloses a method for extracting Polygonatum sibiricum peptide. The preparation method of Polygonatum sibiricum peptide is to pass an electric current through Polygonatum sibiricum stock solution to obtain a precipitate, and then prepare Polygonatum sibiricum peptide by a certain method. Whether the method of passing an electric current can improve the preparation efficiency and whether the activity of the peptide can be guaranteed remains to be discussed. CN 108034682 A discloses a process for extracting Polygonatum sibiricum entity small molecule peptide, which process not only includes dual-frequency composite ultrasound and microwave radiation alternating auxiliary extraction, but also includes supercritical CO2 high-pressure extraction. The extraction conditions are relatively extreme and fierce. CN 116649571 A discloses a Polygonatum odoratum peptide oral liquid and a preparation method thereof. However, according to the preparation method in the patent application, the obtained extract should be a mixture of Polygonatum odoratum polysaccharide and Polygonatum odoratum polypeptide, and the polysaccharide is the main component, with less protein and peptides. Some studies have conducted microwave extraction and antioxidant studies on Polygonatum odoratum glycoprotein. The yield of Polygonatum odoratum glycoprotein obtained is 8.85%. Studies have shown that Polygonatum odoratum glycoprotein has significant reducing power and the ability to scavenge DPPH free radicals. However, glycoprotein is a type of macromolecular complex composed of two substances, sugar and protein, connected in a certain way, which is quite different from protein peptides with smaller molecular weight.

[0007] In summary, the current preparation technology of Chinese medicine protein peptides and their application in the cosmetics field face several challenges. On the one hand, the harsh conditions commonly used in the preparation process, such as high temperature, high pressure, acid-base hydrolysis and electricity, often damage the natural structure and biological activity of protein peptides, thereby weakening their skin care effects; on the other hand, most Chinese medicine protein peptide products on the market contain many impurities and are not pure enough; furthermore, the functional limitations of a single Chinese medicine protein peptide are large, making it difficult to meet consumers' expectations for the multifunctionality of cosmetics. In addition, the safety of cosmetic raw materials has always been the focus of attention in the industry. Ensuring that Chinese medicine protein peptides are both safe and non-irritating is a key issue that needs to be overcome.

[0008] In view of the above background, it is of great significance to provide a multifunctional Chinese herbal protein peptide composition. At the same time, it is particularly urgent to develop a Chinese herbal protein peptide composition that can overcome the shortcomings of the existing technology, adopt a mild and environmentally friendly preparation process, and retain the biological activity of the Chinese herbal protein peptide to the greatest extent. Summary of the invention

[0009] The purpose of the present invention is to overcome the above-mentioned defects and deficiencies in the prior art and to provide a traditional Chinese medicine protein peptide composition with whitening, anti-aging and moisturizing effects.

[0010] The second object of the present invention is to provide a method for preparing the traditional Chinese medicine protein peptide composition.

[0011] The third object of the present invention is to provide application of the traditional Chinese medicine protein peptide composition in cosmetics.

[0012] The above-mentioned object of the present invention is achieved through the following technical solutions:

[0013] The present invention firstly provides a traditional Chinese medicine protein peptide composition, which is composed of yellow spherical protein peptide and polygonatum protein peptide in a mass ratio of 1 to 3:1; the preparation method of the yellow spherical protein peptide and polygonatum protein peptide is that a composite enzyme is used for enzymatic hydrolysis to obtain crude protein, and the crude protein is then enzymatically hydrolyzed twice with a composite alkaline protein and pepsin in sequence to obtain small molecule peptides with a molecular weight below 10KDa, the composite enzyme is α-mannosidase, β-galactosidase, α-glucosidase and β-glucosidase, and the composite alkaline protein is trypsin and papain.

[0014] The present invention proposes a scientifically proportioned Chinese medicine protein peptide composition, and obtains yellow spherical protein peptide and yuzhu protein peptide by three enzymatic hydrolysis methods. Specifically, the first enzymatic hydrolysis mainly aims to hydrolyze glycoprotein and decompose glycoprotein into sugar and protein, and the second enzymatic hydrolysis mainly aims to decompose macromolecular protein into smaller peptide segments. The third enzymatic hydrolysis is based on the second enzymatic hydrolysis, and further decomposes medium-sized peptide segments into small-molecule peptides. The protein peptide obtained by this preparation process has a high purity, and the content of yellow spherical protein peptide ranges from 83.6% to 87.3%, and the content of yuzhu protein peptide ranges from 85.5% to 88.1%. Further, yellow spherical protein peptide and yuzhu protein peptide are combined in a specific ratio, and it is found that the combination of the two protein peptides has a synergistic effect, which significantly improves the multiple skin care effects such as whitening, anti-aging, and moisturizing. Specifically, at the same dosage, the yellow spherical protein peptide and yuzhu protein peptide composition of a specific ratio have higher whitening, anti-aging and moisturizing effects than a single protein peptide.

[0015] Furthermore, the preparation method of the polyanthocyanin peptide and the polyanthocyanin peptide comprises the following steps:

[0016] S1. Performing hydrothermal extraction on the Chinese medicine raw materials to obtain a Chinese medicine extract;

[0017] S2. The Chinese medicine extract is initially hydrolyzed using a composite enzyme to obtain crude protein of Chinese medicine; the composite enzyme is α-mannosidase, β-galactosidase, α-glucosidase and β-glucosidase;

[0018] S3. The crude protein solution of traditional Chinese medicine is hydrolyzed for a second time using a composite alkaline protease to obtain a second hydrolyzate; the composite alkaline protease is trypsin and papain;

[0019] S4. performing a third enzymatic hydrolysis on the second enzymatic hydrolyzate using pepsin to obtain a third enzymatic hydrolyzate;

[0020] S5. The third enzymatic hydrolysate was ultrafiltered and post-treated using a membrane with a molecular weight cutoff of 10 KDa to obtain a Chinese medicine protein peptide;

[0021] The Chinese medicinal raw material is Polygonatum odoratum or Polygonatum sibiricum.

[0022] Therefore, the present invention optimizes the preparation process and adopts three enzymatic hydrolysis to prepare protein peptides, thereby avoiding the destruction of components by extreme conditions. The operation is simple, gentle and environmentally friendly, which helps to maintain the biological activity of the protein peptides and improve the purity of the protein peptides. The obtained protein peptides are safe, non-irritating and have high purity.

[0023] Furthermore, step S1 is to crush the Chinese medicine raw material, immerse the polygonatum powder in distilled water according to a material-liquid mass ratio of 1:13-17, and boil for 2-2.5 hours to obtain a Chinese medicine extract.

[0024] Preferably, step S1 is to crush the Chinese medicine raw material, immerse the polygonatum powder in distilled water according to a material-liquid mass ratio of 1:15, and boil for 2 hours to obtain a Chinese medicine extract.

[0025] Furthermore, step S2 is to adjust the pH value of the extract to 6.0-8.0, add complex enzyme, perform enzymolysis at 50°C for 3-7 hours to obtain the first enzymolysis solution, and inactivate the enzyme; centrifuge the first enzymolysis solution to obtain the supernatant, and precipitate with alcohol to obtain the crude protein of traditional Chinese medicine.

[0026] Preferably, the mass ratio of α-mannosidase, β-galactosidase, α-glucosidase and β-glucosidase in the complex enzyme is 0.8-1.2: 0.8-1.2: 0.8-1.2: 0.8-1.2.

[0027] Preferably, the mass ratio of α-mannosidase, β-galactosidase, α-glucosidase and β-glucosidase in the complex enzyme is 1:1:1:1.

[0028] Furthermore, step S3 is to mix the crude protein of traditional Chinese medicine with distilled water in a mass ratio of 1:13-17, stir evenly, heat to 45-50°C, add composite alkaline protease, adjust the pH value to 7.0-9.0, stir and hydrolyze for 2.0-6.0 hours to obtain a second enzymolysis solution.

[0029] Preferably, step S3 is to mix the crude protein of traditional Chinese medicine with distilled water in a mass ratio of 1:15, stir evenly, heat to 50° C., add composite alkaline protease, adjust the pH value to 7.0-9.0, stir and hydrolyze for 2.0-6.0 hours to obtain a second enzymolysis solution.

[0030] Preferably, the mass ratio of trypsin to papain in the composite alkaline protease is 0.8-1.2:0.8-1.2.

[0031] Preferably, the mass ratio of trypsin to papain in the composite alkaline protease is 1:1.

[0032] Furthermore, step S4 is to add pepsin to the second enzymolysis solution, adjust the pH to 2.0-3.5, perform a third enzymolysis, the enzymolysis time is 2.0-6.0 hours, inactivate the enzyme, and obtain a third enzymolysis solution.

[0033] Furthermore, when the Chinese medicine raw material is Polygonatum sibiricum, the amount of the composite enzyme in step S2 is 1-3% of the mass of the Chinese medicine extract; the amount of the composite alkaline protease in step S3 is 1-5% of the mass of the Chinese medicine crude protein solution; the amount of pepsin in step S4 is 1-5% of the mass of the second enzymatic hydrolyzate.

[0034] Furthermore, when the Chinese medicine raw material is Polygonatum odoratum, the amount of the complex enzyme in step S2 is 1.5-3.5% of the mass of the Chinese medicine extract; the amount of the complex alkaline protease in step S3 is 1.5-5.5% of the mass of the Chinese medicine crude protein solution; the amount of pepsin in step S4 is 1.5-5.5% of the mass of the second enzymatic hydrolyzate.

[0035] The present invention provides application of the traditional Chinese medicine protein peptide composition in preparing whitening, anti-aging and moisturizing cosmetics.

[0036] Furthermore, the cosmetic is an aqueous solution, emulsion, cream or essence.

[0037] The Chinese medicine protein peptide composition is not only suitable for cosmetics in various dosage forms such as aqueous solutions, emulsions, creams, essences, etc., but also provides the cosmetics industry with a new, multifunctional and non-irritating raw material option, fully meeting consumers' urgent needs for natural, safe and environmentally friendly cosmetics.

[0038] The present invention provides a cosmetic comprising the above-mentioned traditional Chinese medicine protein peptide composition.

[0039] Furthermore, the mass percentage of the traditional Chinese medicine protein peptide composition in the cosmetics is 0.1-5%.

[0040] Compared with the prior art, the present invention has the following beneficial effects:

[0041] The present invention provides a Chinese medicine protein peptide composition, which is composed of yellow spherical protein peptide and polygonatum protein peptide, wherein the yellow spherical protein peptide and polygonatum protein peptide are both protein peptides with a molecular weight of less than 10KDa obtained by three enzymatic hydrolysis methods. The Chinese medicine protein peptide composition is obtained by two raw materials in an appropriate ratio, and the Chinese medicine protein peptide composition not only has a variety of skin care effects of whitening, anti-aging and moisturizing, but also the two protein peptide raw material components act synergistically, and the effect is significantly better than using only one of them. The present invention also provides a preparation method of the yellow spherical protein peptide and polygonatum protein peptide, which optimizes the preparation process through multiple enzymatic hydrolysis technology, realizes a mild and environmentally friendly production process, and maximizes the biological activity of the Chinese medicine protein peptide, while improving the purity of the protein peptide to ensure non-irritation, and the preparation method is simple to operate, mild conditions, environmentally friendly, and easy to produce. Compared with most chemical synthetic raw materials used in cosmetics, this Chinese medicine protein peptide composition has the advantages of being non-irritating to the skin, mild and safe, and having no toxic side effects. It is used in cosmetics in different dosage forms such as water-based solutions, emulsions, creams, essences, etc., and has a wide range of uses. It provides the cosmetics industry with a multifunctional, non-irritating, natural and mild raw material option, meeting consumers' urgent needs for safe, efficient and environmentally friendly cosmetics. DETAILED DESCRIPTION

[0042] The present invention is further described with specific examples, but the examples do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0043] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0044] α-Mannosidase (Product No.: M923745-1mg; ≥15units / mg protein (biuret)), β-galactosidase (Product No.: S10066-1KU; purity / concentration: BR, 250-600u / mg protein), α-glucosidase (Product No.: S10050-1ku; purity level: 50units / mg protein yeast), β-glucosidase (Product No.: G824159-10mg; purity / concentration: 10-30U / mg, source: almond), trypsin (Product No.: S10034-100g; purity / concentration: BR; 1:250), papain (Product No.: P815680-500g; purity level: ≥3units / mg), pepsin (Product No.: 9001-75-6; specification: >3000U / mg).

[0045] Example 1

[0046] (1) Preparation of yellow protamine peptide

[0047] S1. Wash and dry the polygonatum sibiricum, grind it and pass it through a 120-mesh sieve, immerse the polygonatum sibiricum powder in distilled water at a material-liquid mass ratio of 1:15, stir evenly, and boil it at 90°C for 2 hours to obtain a polygonatum sibiricum extract.

[0048] S2. The pH value of the extract is adjusted to 6.0, and a composite enzyme (α-mannosidase, β-galactosidase, α-glucosidase, β-glucosidase, the mass ratio of the four is 1:1:1:1) equivalent to 1.0% of the mass of the mixed solution is added thereto, and the enzymolysis is carried out at a temperature of 50°C for 3.0 hours to obtain the first enzymolysis solution, which is heated at 90°C for 5 minutes to inactivate the enzyme and terminate the reaction. The first enzymolysis solution is centrifuged at a speed of 6000rpm for 20 minutes, and the supernatant is taken. The temperature is adjusted to 20°C, 70% of the volume of ethanol is added to the supernatant, and it is stirred evenly. The mixed solution is placed under refrigeration at 4°C and allowed to stand for 24 hours. Centrifuge at a speed of 3500rpm for 10 minutes to obtain the precipitate, and the crude protein of Polygonatum sibiricum is obtained.

[0049] S3. Mix the crude protein of Polygonatum sibiricum with distilled water in a mass ratio of 1:15, stir evenly, heat to 50°C, add a composite alkaline protease (a mixture of trypsin and papain, with a mass ratio of 1:1) equivalent to 1.0% of the mass of the protein solution, adjust the pH value to 7.0, stir and hydrolyze for 2.0 hours to obtain the second enzymolysis solution.

[0050] S4. After the enzymolysis is completed, pepsin equivalent to 1.0% of the solution mass is added to the second enzymolysis solution, the pH is adjusted to 2.0, and the third enzymolysis is performed for 2.0 hours. The third enzymolysis solution is heated to 90°C, the enzyme is inactivated for 15 minutes, and the reaction is terminated.

[0051] S5. The enzyme-killed hydrolyzate is purified by centrifugation at a speed of 15000 r / min to obtain a centrifuge. The centrifuge is filtered through an ultrafiltration membrane with a molecular weight cutoff of 10 KDa, and the filtrate is concentrated and freeze-dried to obtain polyanthocyanin peptide.

[0052] (2) Preparation of Polygonatum odoratum peptide

[0053] Referring to the preparation method of the above-mentioned yellow protamine peptide, the differences are as follows: S2 is a composite enzyme (α-mannosidase, β-galactosidase, α-glucosidase, β-glucosidase, the mass ratio of the four is 1:1:1:1) equivalent to 1.5% of the mass of the mixed solution; S3 is a composite alkaline protease (a mixture of trypsin and papain, the mass ratio is 1:1) equivalent to 1.5% of the mass of the protein solution; S4 is a pepsin equivalent to 1.5% of the mass of the solution.

[0054] (3) Preparation of Chinese medicine protein peptide composition

[0055] The frozen powders of Polygonatum protein peptide and Polygonatum protein peptide are mixed evenly in a weight ratio of 1:1 to obtain a Chinese medicine protein peptide composition.

[0056] Example 2

[0057] (1) Preparation of yellow protamine peptide

[0058] S1. Wash and dry the polygonatum sibiricum, grind it and pass it through a 120-mesh sieve, immerse the polygonatum sibiricum powder in distilled water at a material-liquid mass ratio of 1:15, stir evenly, and boil it at 90°C for 2 hours to obtain a polygonatum sibiricum extract.

[0059] S2. The pH value of the extract is adjusted to 6.5, and a composite enzyme (α-mannosidase, β-galactosidase, α-glucosidase, β-glucosidase, the mass ratio of the four is 1:1:1:1) equivalent to 1.5% of the mass of the mixed solution is added thereto, and the enzymolysis is carried out at a temperature of 50°C for 4.0 hours to obtain the first enzymolysis solution, which is heated at 90°C for 5 minutes to inactivate the enzyme and terminate the reaction. The first enzymolysis solution is centrifuged at a speed of 6000rpm for 20 minutes, and the supernatant is taken. The temperature is adjusted to 20°C, 70% of the volume of ethanol is added to the supernatant, stirred evenly, and the mixed solution is placed under refrigeration at 4°C for 24 hours. Centrifuge at a speed of 3500rpm for 10 minutes to obtain the precipitate, and the crude protein of Polygonatum sibiricum is obtained.

[0060] S3. Mix the crude protein of Polygonatum sibiricum with distilled water in a mass ratio of 1:15, stir evenly, heat to 50°C, add a composite alkaline protease (a mixture of trypsin and papain, with a mass ratio of 1:1) equivalent to 2.0% of the mass of the protein solution, adjust the pH value to 7.5, stir and hydrolyze for 3.0 hours to obtain the second enzymolysis solution.

[0061] S4. After the enzymolysis is completed, pepsin equivalent to 2.0% of the solution mass is added to the second enzymolysis solution, the pH is adjusted to 2.5, and the third enzymolysis is performed for 3.0 hours. The third enzymolysis solution is heated to 90°C, the enzyme is inactivated for 15 minutes, and the reaction is terminated.

[0062] S5. The enzyme-killed hydrolyzate is purified by centrifugation at a speed of 15000 r / min to obtain a centrifuge. The centrifuge is filtered through an ultrafiltration membrane with a molecular weight cutoff of 10 KDa, and the filtrate is concentrated and freeze-dried to obtain polyanthocyanin peptide.

[0063] (2) Preparation of Polygonatum odoratum peptide

[0064] Referring to the above-mentioned method for preparing yellow protamine peptide, the differences are as follows: S2 is a composite enzyme (α-mannosidase, β-galactosidase, α-glucosidase, β-glucosidase, the mass ratio of the four is 1:1:1:1) equivalent to 2.0% of the mass of the mixed solution; S3 is a composite alkaline protease (a mixture of trypsin and papain, the mass ratio is 1:1) equivalent to 2.5% of the mass of the protein solution; S4 is a pepsin equivalent to 2.5% of the mass of the solution added to the second enzymatic hydrolysis solution.

[0065] (3) Preparation of Chinese medicine protein peptide composition

[0066] The frozen powders of Polygonatum protein peptide and Polygonatum odoratum protein peptide are uniformly mixed in a weight ratio of 1.5:1 to obtain a traditional Chinese medicine protein peptide composition.

[0067] Example 3

[0068] (1) Preparation of yellow protamine peptide

[0069] S1. Wash and dry the polygonatum sibiricum, grind it and pass it through a 120-mesh sieve, immerse the polygonatum sibiricum powder in distilled water at a material-liquid mass ratio of 1:15, stir evenly, and boil it at 90°C for 2 hours to obtain a polygonatum sibiricum extract.

[0070] S2. The pH value of the extract is adjusted to 7.0, and a composite enzyme (α-mannosidase, β-galactosidase, α-glucosidase, β-glucosidase, the mass ratio of the four is 1:1:1:1) equivalent to 2.0% of the mass of the mixed solution is added thereto, and the enzymolysis is carried out at a temperature of 50°C for 5.0 hours to obtain the first enzymolysis solution, which is heated at 90°C for 5 minutes to inactivate the enzyme and terminate the reaction. The first enzymolysis solution is centrifuged at a speed of 6000rpm for 20 minutes, and the supernatant is taken. The temperature is adjusted to 20°C, 70% of the volume of ethanol is added to the supernatant, stirred evenly, and the mixed solution is placed under refrigeration at 4°C for 24 hours. Centrifuge at a speed of 3500rpm for 10 minutes to obtain the precipitate, and the crude protein of Polygonatum sibiricum is obtained.

[0071] S3. Mix the crude protein of Polygonatum sibiricum with distilled water in a mass ratio of 1:15, stir evenly, heat to 50°C, add a composite alkaline protease (a mixture of trypsin and papain, with a mass ratio of 1:1) equivalent to 3.0% of the mass of the protein solution, adjust the pH value to 8.0, stir and hydrolyze for 4.0 hours to obtain the second enzymolysis solution.

[0072] S4. After the enzymolysis is completed, pepsin equivalent to 3.0% of the solution mass is added to the second enzymolysis solution, the pH is adjusted to 2.8, and the third enzymolysis is performed for 4.0 hours. The third enzymolysis solution is heated to 90°C, the enzyme is inactivated for 15 minutes, and the reaction is terminated.

[0073] S5. The enzyme-killed hydrolyzate is purified by centrifugation at a speed of 15000 r / min to obtain a centrifuge. The centrifuge is filtered through an ultrafiltration membrane with a molecular weight cutoff of 10 KDa, and the filtrate is concentrated and freeze-dried to obtain polyanthocyanin peptide.

[0074] (2) Preparation of Polygonatum odoratum peptide

[0075] Referring to the preparation method of the above-mentioned yellow protamine peptide, the differences are as follows: S2 is a composite enzyme (α-mannosidase, β-galactosidase, α-glucosidase, β-glucosidase, the ratio of the four is 1:1:1:1) equivalent to 2.5% of the mass of the mixed solution; S3 is a composite alkaline protease (a mixture of trypsin and papain, the mass ratio is 1:1) equivalent to 3.5% of the mass of the protein solution; S4 is a pepsin equivalent to 3.5% of the mass of the solution.

[0076] (3) Preparation of Chinese medicine protein peptide composition

[0077] The frozen powders of Polygonatum protein peptide and Polygonatum odoratum protein peptide are uniformly mixed in a weight ratio of 2:1 to obtain a traditional Chinese medicine protein peptide composition.

[0078] Example 4

[0079] (1) Preparation of yellow protamine peptide

[0080] S1. Wash and dry the polygonatum sibiricum, grind it and pass it through a 120-mesh sieve, immerse the polygonatum sibiricum powder in distilled water at a material-liquid mass ratio of 1:15, stir evenly, and boil it at 90°C for 2 hours to obtain a polygonatum sibiricum extract.

[0081] S2. The pH value of the extract is adjusted to 7.5, and a composite enzyme (α-mannosidase, β-galactosidase, α-glucosidase, β-glucosidase, the mass ratio of the four is 1:1:1:1) equivalent to 2.5% of the mass of the mixed solution is added thereto, and the enzymolysis is carried out at a temperature of 50°C for 6.0 hours to obtain the first enzymolysis solution, which is heated at 90°C for 5 minutes to inactivate the enzyme and terminate the reaction. The first enzymolysis solution is centrifuged at a speed of 6000rpm for 20 minutes, and the supernatant is taken. The temperature is adjusted to 20°C, 70% of the volume of ethanol is added to the supernatant, stirred evenly, and the mixed solution is placed under refrigeration at 4°C for 24 hours. Centrifuge at a speed of 3500rpm for 10 minutes to obtain the precipitate, and the crude protein of Polygonatum sibiricum is obtained.

[0082] S3. Mix the crude protein of Polygonatum sibiricum with distilled water in a mass ratio of 1:15, stir evenly, heat to 50°C, add a composite alkaline protease (a mixture of trypsin and papain, with a mass ratio of 1:1) equivalent to 4% of the mass of the protein solution, adjust the pH value to 8.5, stir and hydrolyze for 5.0 hours to obtain the second enzymolysis solution.

[0083] S4. After the enzymolysis is completed, pepsin equivalent to 4.0% of the solution mass is added to the second enzymolysis solution, the pH is adjusted to 3.0, and the third enzymolysis is performed for 5.0 hours. The third enzymolysis solution is heated to 90°C, the enzyme is inactivated for 15 minutes, and the reaction is terminated.

[0084] S5. The enzyme-killed hydrolyzate is purified by centrifugation at a speed of 15000 r / min to obtain a centrifuge. The centrifuge is filtered through an ultrafiltration membrane with a molecular weight cutoff of 10 KDa, and the filtrate is concentrated and freeze-dried to obtain polyanthocyanin peptide.

[0085] (2) Preparation of Polygonatum odoratum peptide

[0086] Referring to the preparation method of the above-mentioned yellow protamine peptide, the differences are as follows: S2 is a composite enzyme (α-mannosidase, β-galactosidase, α-glucosidase, β-glucosidase, the ratio of the four is 1:1:1:1) equivalent to 3.0% of the mass of the mixed solution; S3 is a composite alkaline protease (a mixture of trypsin and papain, the mass ratio is 1:1) equivalent to 4.5% of the mass of the protein solution; S4 is a pepsin equivalent to 4.5% of the mass of the solution.

[0087] (3) Preparation of Chinese medicine protein peptide composition

[0088] The frozen powders of Polygonatum protein peptide and Polygonatum odoratum protein peptide are uniformly mixed in a weight ratio of 2.5:1 to obtain a traditional Chinese medicine protein peptide composition.

[0089] Example 5

[0090] (1) Preparation of yellow protamine peptide

[0091] S1. Wash and dry the polygonatum sibiricum, grind it and pass it through a 120-mesh sieve, immerse the polygonatum sibiricum powder in distilled water at a material-liquid mass ratio of 1:15, stir evenly, and boil it at 90°C for 2 hours to obtain a polygonatum sibiricum extract.

[0092] S2. The pH value of the extract is adjusted to 8.0, and a composite enzyme (α-mannosidase, β-galactosidase, α-glucosidase, β-glucosidase, the mass ratio of the four is 1:1:1:1) equivalent to 3.0% of the mass of the mixed solution is added thereto, and the enzymolysis is carried out at a temperature of 50°C for 7.0 hours to obtain the first enzymolysis solution, which is heated at 90°C for 5 minutes to inactivate the enzyme and terminate the reaction. The first enzymolysis solution is centrifuged at a speed of 6000rpm for 20 minutes, and the supernatant is taken. The temperature is adjusted to 20°C, 70% of the volume of ethanol is added to the supernatant, stirred evenly, and the mixed solution is placed under refrigeration at 4°C for 24 hours. Centrifuge at a speed of 3500rpm for 10 minutes to obtain the precipitate, and the crude protein of Polygonatum sibiricum is obtained.

[0093] S3. Mix the crude protein of Polygonatum sibiricum with distilled water in a mass ratio of 1:15, stir evenly, heat to 50°C, add a composite alkaline protease (a mixture of trypsin and papain, with a mass ratio of 1:1) equivalent to 5.0% of the mass of the protein solution, adjust the pH value to 9.0, stir and hydrolyze for 6.0 hours to obtain the second enzymolysis solution.

[0094] S4. After the enzymolysis is completed, pepsin equivalent to 5.0% of the solution mass is added to the second enzymolysis solution, the pH is adjusted to 3.5, and the third enzymolysis is performed for 6.0 hours. The third enzymolysis solution is heated to 90°C, the enzyme is inactivated for 15 minutes, and the reaction is terminated.

[0095] S5. The enzyme-killed hydrolyzate is purified by centrifugation at a speed of 15000 r / min to obtain a centrifuge. The centrifuge is filtered through an ultrafiltration membrane with a molecular weight cutoff of 10 KDa, and the filtrate is concentrated and freeze-dried to obtain polyanthocyanin peptide.

[0096] (2) Preparation of Polygonatum odoratum peptide

[0097] Referring to the preparation method of the above-mentioned yellow protamine peptide, the difference is that: S2 is a composite enzyme (α-mannosidase, β-galactosidase, α-glucosidase, β-glucosidase, the ratio of the four is 1:1:1:1) equivalent to 3.5% of the mass of the mixed solution; S3 is a composite alkaline protease (a mixture of trypsin and papain, the mass ratio is 1:1) equivalent to 5.5% of the mass of the protein solution; S4 is a pepsin equivalent to 5.5% of the mass of the solution.

[0098] (3) Preparation of Chinese medicine protein peptide composition

[0099] The frozen powders of Polygonatum protein peptide and Polygonatum odoratum protein peptide are uniformly mixed in a weight ratio of 3:1 to obtain a traditional Chinese medicine protein peptide composition.

[0100] Comparative Example 1

[0101] The difference between Comparative Example 1 and Example 3 is that, in the preparation process of yellow protamine peptide and polygonatum protein peptide, only composite enzyme (α-mannosidase, β-galactosidase, α-glucosidase, β-glucosidase) and composite alkaline protease are used for two enzymatic hydrolysis, and pepsin is not used for the third enzymatic hydrolysis, that is, step S4 is omitted, and other preparation conditions and the dosage ratio of yellow protamine peptide and polygonatum protein peptide in the traditional Chinese medicine protein peptide composition are consistent with Example 3.

[0102] Comparative Example 2

[0103] The difference between Comparative Example 2 and Example 3 is that, in the preparation process of yellow protamine peptide and polygonatum protein peptide, only composite enzyme (α-mannosidase, β-galactosidase, α-glucosidase, β-glucosidase) and pepsin are used for enzymatic hydrolysis twice, and composite alkaline protease is not used for enzymatic hydrolysis, that is, step S3 is omitted, and other preparation conditions and the dosage ratio of yellow protamine peptide and polygonatum protein peptide in the traditional Chinese medicine protein peptide composition are consistent with Example 3.

[0104] Comparative Example 3

[0105] The difference between Comparative Example 3 and Example 3 is that, in the preparation process of yellow spherical protein peptide and polygonatum protein peptide, only composite alkaline protease and pepsin are used for enzymatic hydrolysis twice, and composite enzyme (α-mannosidase, β-galactosidase, α-glucosidase, β-glucosidase) is not used for enzymatic hydrolysis, that is, step S2 is omitted, and other preparation conditions and the dosage ratio of yellow spherical protein peptide and polygonatum protein peptide in the traditional Chinese medicine protein peptide composition are consistent with those in Example 3.

[0106] Comparative Example 4

[0107] The difference between Comparative Example 4 and Example 3 is that, in the preparation process of yellow spherical protein peptide and polygonatum protein peptide, only composite enzyme (α-mannosidase, β-galactosidase, α-glucosidase, β-glucosidase) is used for enzymatic hydrolysis, and composite alkaline protease and pepsin are not used for enzymatic hydrolysis, that is, steps S3 and S4 are omitted, and other preparation conditions and the dosage ratio of yellow spherical protein peptide and polygonatum protein peptide in the traditional Chinese medicine protein peptide composition are consistent with those in Example 3.

[0108] Comparative Example 5

[0109] The difference between Comparative Example 5 and Example 3 is that, in the preparation process of yellow protamine peptide and polygonatum protein peptide, only composite alkaline protease is used for enzymatic hydrolysis, and composite enzyme (α-mannosidase, β-galactosidase, α-glucosidase, β-glucosidase) and pepsin are not used for enzymatic hydrolysis, that is, steps S2 and S4 are omitted, and other preparation conditions and the dosage ratio of yellow protamine peptide and polygonatum protein peptide in the traditional Chinese medicine protein peptide composition are consistent with those in Example 3.

[0110] Comparative Example 6

[0111] The difference between Comparative Example 6 and Example 3 is that, in the preparation process of yellow protamine peptide and polygonatum protein peptide, only pepsin is used for enzymatic hydrolysis, and composite enzymes (α-mannosidase, β-galactosidase, α-glucosidase, β-glucosidase) and composite alkaline protease are not used for enzymatic hydrolysis, that is, steps S2 and S3 are omitted, and other preparation conditions and the dosage ratio of yellow protamine peptide and polygonatum protein peptide in the traditional Chinese medicine protein peptide composition are consistent with those in Example 3.

[0112] Comparative Example 7

[0113] The difference between Comparative Example 7 and Example 3 is that the dosage ratio of the raw materials in the Chinese medicine protein peptide composition is different, the dosage ratio of Polygonatum protein peptide and Polygonatum protein peptide is 1:5, but the total amount is the same as that of Example 3, and the preparation process is consistent with that of Example 3.

[0114] Comparative Example 8

[0115] The difference between Comparative Example 8 and Example 3 is that the dosage ratio of the raw materials in the Chinese medicine protein peptide composition is different, the dosage ratio of Polygonatum protein peptide and Polygonatum protein peptide is 1:3, but the total amount is the same as that of Example 3, and the preparation process is consistent with that of Example 3.

[0116] Comparative Example 9

[0117] The difference between Comparative Example 9 and Example 3 is that the dosage ratio of the raw materials in the Chinese medicine protein peptide composition is different, the dosage ratio of Polygonatum protein peptide and Polygonatum protein peptide is 5:1, but the total amount is the same as that of Example 3, and the preparation process is consistent with that of Example 3.

[0118] Comparative Example 10

[0119] The difference between Comparative Example 10 and Example 3 is that the dosage ratio of the raw materials in the Chinese medicine protein peptide composition is different, the dosage ratio of Polygonatum protein peptide and Polygonatum protein peptide is 7:1, but the total amount is the same as that of Example 3, and the preparation process is consistent with that of Example 3.

[0120] Comparative Example 11

[0121] The difference between Comparative Example 11 and Example 3 is that the Chinese medicine protein peptide composition contains only Polygonatum protein peptide, and no Polygonatum protein peptide is added, but the total amount is the same as that of Example 3, and the preparation process of Polygonatum protein peptide is the same as that of Example 3.

[0122] Comparative Example 12

[0123] The difference between Comparative Example 12 and Example 3 is that the Chinese medicine protein peptide composition contains only Polygonatum odoratum protein peptide, and no Polygonatum odoratum protein peptide is added, but the total amount is the same as that of Example 3, and the preparation process of Polygonatum odoratum protein peptide is the same as that of Example 3.

[0124] Application Example 1 A water-based preparation containing a Chinese medicine protein peptide composition and a preparation method thereof

[0125] Raw materials (mass fractions): 0.1 parts of Chinese medicine protein peptide composition, 3 parts of propylene glycol, 0.1 parts of sodium hyaluronate, 3 parts of ethanol, 3 parts of glycerol, 0.1 parts of EDTA-2Na, appropriate amount of citric acid, adjust the pH range (5.5-6.5), 0.4 parts of p-hydroxyacetophenone, 0.4 parts of phenoxyethanol, 0.02 parts of flavor, add deionized water to 100 parts.

[0126] Preparation steps:

[0127] Weigh all ingredients accurately to ensure that the proportion of each ingredient meets the recipe requirements.

[0128] Add an appropriate amount of deionized water into a stirring tank, slowly add the Chinese medicine protein peptide composition, sodium hyaluronate, and glycerin into the water, and continue stirring until they are completely dissolved to obtain a main solution.

[0129] Take another clean container, add propylene glycol, ethanol and flavor, and stir until the flavor is completely dissolved to obtain a flavor solution.

[0130] Slowly add the prepared flavor solution into the main solution and stir until the two solutions are completely mixed and there is no separation.

[0131] Weigh an appropriate amount of citric acid and slowly add it to the stirring tank, checking the pH value with a pH meter while adding until the desired pH range (5.5-6.5) is reached.

[0132] Add EDTA-2Na to the mixing tank and stir until dissolved.

[0133] Add p-hydroxyacetophenone and phenoxyethanol to the mixing tank and continue to stir evenly. Finally, add deionized water to 100 parts, and perform final stirring and adjustment to ensure that all components of the formula are fully mixed and evenly mixed to obtain an aqueous solution containing a Chinese medicine protein peptide composition.

[0134] Application Example 2 An emulsion containing a Chinese medicine protein peptide composition and its preparation method

[0135] Raw materials (mass fractions): 1 part of Chinese medicine protein peptide composition, 2 parts of glycerin, 0.1 parts of sodium hyaluronate, 0.1 parts of allantoin, 0.02 parts of essence, 10 parts of sweet almond oil, 8 parts of jojoba oil, 2 parts of beeswax, 0.5 parts of vitamin E, 5 parts of Span 80, 0.5 parts of carbomer, 0.5 parts of phenoxyethanol, appropriate amount of citric acid, and deionized water added to 100 parts.

[0136] Preparation steps:

[0137] Weighing raw materials: Accurately weigh all raw materials to ensure that the proportion of each raw material meets the recipe requirements.

[0138] Oil phase preparation: Mix sweet almond oil, jojoba oil, beeswax and vitamin E in a container, heat to about 70-75°C, melt them completely and mix them evenly.

[0139] Preparation of water phase: In a separate container, heat an appropriate amount of deionized water to about 70-75°C. Then, add glycerin, sodium hyaluronate, allantoin and carbomer in sequence, stirring while adding until completely dissolved.

[0140] Emulsification: When the water phase is still warm, slowly add Span 80 and stir vigorously to fully emulsify it. Slowly pour the preheated oil phase into the water phase, stirring quickly with a stirrer to form a preliminary emulsion. Use a homogenizer to homogenize the preliminary emulsion to improve the stability and fineness of the emulsion.

[0141] Cooling adjustment: Cool the homogenized emulsion to room temperature and stir appropriately to prevent stratification. When the emulsion cools to near room temperature, add fragrance and phenoxyethanol and stir evenly. Add appropriate amount of citric acid to adjust the pH value to 5.5-6.5.

[0142] Quality testing: The emulsion is subjected to quality tests such as appearance, odor, and stability to ensure that the product meets the standard requirements, that is, the emulsion containing the Chinese medicine protein peptide composition is obtained.

[0143] Application Example 3 A cream containing a Chinese medicine protein peptide composition and its preparation method

[0144] Raw materials (mass fraction): 3 parts of Chinese medicine protein peptide composition, 5 parts of trehalose, 0.3 parts of sodium hyaluronate, 0.1 parts of allantoin, 12 parts of rosehip oil, 10 parts of sweet almond oil, 2 parts of candelilla wax, 2.5 parts of cetearyl alcohol emulsifier, 0.4 parts of vitamin E, 2 parts of Polysorbate 80, 0.3 parts of phenoxyethanol, appropriate amount of citric acid, deionized water added to 100 parts

[0145] Preparation steps:

[0146] Weighing raw materials: Accurately weigh all raw materials to ensure that the proportion of each raw material meets the recipe requirements.

[0147] Oil phase preparation: Mix rosehip oil, sweet almond oil, candelilla wax, vitamin E and cetearyl alcohol emulsifier in a container and heat to 85°C until all solid ingredients are completely melted and mixed evenly.

[0148] Preparation of aqueous phase: In another container, add appropriate amount of deionized water and heat to 85°C, then add trehalose, sodium hyaluronate and allantoin in sequence, stirring while adding until completely dissolved.

[0149] Emulsification: When the water phase is still warm, add Polysorbate 80 to the water phase, stirring while adding to fully dissolve it. Slowly pour the preheated oil phase into the water phase, stirring quickly with an electric stirrer while pouring to form a preliminary cream. Use a homogenizer to homogenize the preliminary formed cream to improve the stability and fineness of the cream.

[0150] Cooling adjustment: Cool the emulsified cream to room temperature, stirring appropriately to prevent stratification. When the cream cools to near room temperature, add phenoxyethanol and stir evenly. Check the pH value of the cream and add appropriate amount of citric acid to adjust the pH value to 5.5-6.5.

[0151] Quality testing: The cream is subjected to quality tests such as appearance, smell, and stability to ensure that the product meets the standard requirements, that is, the cream containing the Chinese medicine protein peptide composition is obtained.

[0152] Application Example 4: An essence containing a Chinese medicine protein peptide composition and its preparation method

[0153] Raw materials (mass fractions): 5 parts of Chinese medicine protein peptide composition, 0.5 parts of sodium hyaluronate, 5 parts of glycerol, 5 parts of butylene glycol, 3 parts of 1,3-propylene glycol, 2 parts of trehalose, 0.1 parts of allantoin, 0.05 parts of disodium EDTA, 0.5 parts of phenoxyethanol, 0.3 parts of carbomer, appropriate amount of triethanolamine, and deionized water added to 100 parts.

[0154] Preparation steps: Add carbomer to an appropriate amount of deionized water, stir evenly to fully swell it, and heat to about 70°C to promote dissolution. In a clean, sterilized container, add deionized water (reserved for the dissolved carbomer), and then add the Chinese medicine protein peptide composition, sodium hyaluronate, glycerol, butylene glycol, 1,3-propylene glycol, trehalose, allantoin and disodium EDTA in sequence. Heat to about 75°C, stirring while heating to ensure that all ingredients are completely dissolved. When the water phase temperature drops to about 70°C, slowly add the pre-prepared carbomer solution, stirring while adding to avoid a large amount of foam. Continue stirring until the carbomer is completely dissolved to form a uniform viscous solution. Use triethanolamine to slowly adjust the pH value of the solution to between about 6.0 and 6.5. When the solution temperature drops to near room temperature, add phenoxyethanol and stir evenly. Use a homogenizer to homogenize the solution to improve the uniformity and stability of the solution. The essence is subjected to quality inspection, including appearance, odor, pH value, viscosity and other indicators, to ensure that the product meets the standard requirements, thus obtaining an essence containing a Chinese medicine protein peptide composition.

[0155] Test Example 1 Determination of the Content of Polygonatum Protein Peptide and Polygonatum Protein Peptide

[0156] The content of the polyanthocyanin peptide and the polyanthocyanin peptide involved in Examples 1 to 5 and Comparative Examples 1 to 6 was determined to evaluate the advantages and disadvantages of the protein peptide extraction and purification method of the present invention. The method and results are as follows:

[0157] 1. Methodology

[0158] According to the provisions of the "National Food Safety Standard Plant Protein Peptides for Food Processing" (GB 31611-2023), the "Kjeldahl Nitrogen Determination Method" in the "National Food Safety Standard Determination of Protein in Food" (GB 5009.5-2016) is used to determine the content of protein peptides.

[0159] 2. Determination principle: The protein peptide is decomposed under catalytic heating conditions, and the generated ammonia combines with sulfuric acid to form ammonium sulfate. Alkalinization distillation frees the ammonia, which is absorbed by boric acid and titrated with sulfuric acid or hydrochloric acid standard titration solution. The nitrogen content is calculated based on the acid consumption, and then multiplied by the conversion coefficient (6.25), which is the content of the protein peptide.

[0160] 3. Materials and Reagents

[0161] The Chinese medicine protein peptide compositions were prepared according to the methods of Examples 1 to 5 and Comparative Examples 1 to 6, and dried and stored for later use.

[0162] Copper sulfate (CuSO4·5H2O), potassium sulfate (K2SO4), sulfuric acid (H2SO4), boric acid (H3BO3), methyl red indicator (C 15 H 15 N3O2), bromocresol green indicator (C 21 H 14 Br4O5S), methylene blue indicator (C 16 H 18 CIN3S·3H2O), sodium hydroxide (NaOH), and 95% ethanol (C2H5OH) were all analytically pure.

[0163] Boric acid solution (20 g / L): Weigh 20 g of boric acid, dissolve it in water and dilute to 1000 mL.

[0164] Sodium hydroxide solution (400 g / L): Weigh 40 g of sodium hydroxide, dissolve it in water, cool it, and dilute it to 100 mL.

[0165] Sulfuric acid standard titration solution [c(1 / 2H2SO4)]: 0.0500mol / L.

[0166] Methyl red ethanol solution (1 g / L): weigh 0.1 g of methyl red, dissolve it in 95% ethanol, and dilute it to 100 mL with 95% ethanol.

[0167] Methylene blue ethanol solution (1 g / L): Weigh 0.1 g of methylene blue, dissolve it in 95% ethanol, and dilute it to 100 mL with 95% ethanol.

[0168] Bromocresol green ethanol solution (1 g / L): Weigh 0.1 g of bromocresol green, dissolve it in 95% ethanol, and dilute it to 100 mL with 95% ethanol.

[0169] Mixed indicator solution: Mix 2 parts of methyl red ethanol solution with 1 part of methylene blue ethanol solution before use.

[0170] 4. Analysis steps

[0171] Sample treatment: Weigh 0.2g of the fully mixed solid sample, accurate to 0.001g, transfer it into a dry 100mL nitrogen determination bottle, add 0.4g copper sulfate, 6g potassium sulfate and 20mL sulfuric acid, shake it gently, place a small funnel at the mouth of the bottle, and place the bottle at an angle of 45° on an asbestos net with small holes. Heat carefully, and after the contents are completely carbonized and the foam stops completely, increase the firepower and keep the liquid in the bottle slightly boiling until the liquid is blue-green and clear and transparent, and continue heating for 0.5h~1h. Remove and cool, carefully add 20mL of water, cool it, transfer it into a 100mL volumetric flask, and wash the nitrogen determination bottle with a small amount of water, add the washing liquid into the volumetric flask, add water to the scale, mix well and set aside. Perform a reagent blank test at the same time.

[0172] Determination: Install the nitrogen distillation apparatus, fill the steam generator to 2 / 3 with water, add a few glass beads, a few drops of methyl red ethanol solution and a few milliliters of sulfuric acid to keep the water acidic, heat and boil the water in the steam generator and keep it boiling.

[0173] Add 10.0mL of boric acid solution and 1 to 2 drops of mixed indicator to the receiving bottle, and insert the lower end of the condenser under the liquid surface. According to the nitrogen content in the sample, accurately draw 2.0mL of sample treatment solution from the small glass cup and inject it into the reaction chamber. Wash the small glass cup with 10mL of water and let it flow into the reaction chamber, then plug the rod-shaped glass stopper. Pour 10.0mL of sodium hydroxide solution into the small glass cup, lift the glass stopper to let it slowly flow into the reaction chamber, immediately cover the glass stopper and seal it with water. Clamp the spiral clamp and start distillation. After distilling for 10 minutes, move the distillate receiving bottle, and the liquid surface leaves the lower end of the condenser, and distill for another 1 minute. Then rinse the outside of the lower end of the condenser with a small amount of water and remove the distillate receiving bottle. Titrate to the end point with sulfuric acid standard titration solution as soon as possible, that is, the color of the solution changes to gray-blue. At the same time, make a reagent blank.

[0174] The protein content in the sample is calculated as follows:

[0175]

[0176] Where:

[0177] X—protein peptide content in the sample, in grams per hundred grams (g / 100g);

[0178] V1—the volume of sulfuric acid or hydrochloric acid standard titrant consumed by the test solution, in milliliters (mL);

[0179] V2—volume of sulfuric acid or hydrochloric acid standard titrant consumed by reagent blank, in milliliters (mL);

[0180] c—concentration of sulfuric acid standard titration solution, in moles per liter (mol / L);

[0181] The mass of nitrogen equivalent to 0.0140-1.0mL sulfuric acid standard titration solution, in grams (g);

[0182] m—mass of the sample, in grams (g);

[0183] V3—the volume of digestive fluid absorbed, in milliliters (mL);

[0184] 6.25—Nitrogen conversion factor for protein peptides;

[0185] 100—Conversion factor.

[0186] Table 1 Protein peptide content in Examples and Comparative Examples

[0187]

[0188]

[0189] By analyzing the content determination results of the polyanthocyanin peptide and the polyanthocyanin peptide involved in Examples 1 to 5 and Comparative Examples 1 to 6 (Table 1), the following conclusions can be drawn:

[0190] (1) The protein peptide extraction and purification method of the embodiment has high effectiveness and superiority. According to the measurement results, the content of the yellow spherical protein peptide in the embodiment ranges from 84.2% to 87.3%, and the content of the yuzhu protein peptide ranges from 85.5% to 88.1%, which are significantly higher than the content in the comparative example (the content of the yellow spherical protein peptide in the comparative example ranges from 15.4% to 69.1%, and the content of the yuzhu protein peptide ranges from 16.5% to 72.6%). This significant difference shows that the protein peptide extraction and purification method adopted by the present invention has high effectiveness and superiority. Further analysis found that the main difference between the embodiment and the comparative example in the extraction and purification process is the number of enzymatic hydrolysis. The embodiment uses three enzymatic hydrolysis, while the comparative example uses one or two enzymatic hydrolysis. This shows that the number of enzymatic hydrolysis is an important factor affecting the content of protein peptides. By increasing the number of enzymatic hydrolysis, protein peptides can be extracted and purified more effectively.

[0191] (2) The protein peptide extraction and purification method of the embodiment has good stability and reproducibility. The data in the embodiment show that although there are certain fluctuations in the protein peptide content between the embodiments, overall, the fluctuation range is relatively small. The content fluctuation range of Polygonatum sibiricum protein peptide is between 84.2% and 87.3%, and the content fluctuation range of Polygonatum odoratum protein peptide is between 85.5% and 88.1%. This smaller fluctuation range shows that the extraction and purification method has good stability and reproducibility, and can maintain relatively consistent protein peptide extraction effects in different batches.

[0192] In summary, the protein peptide extraction and purification method adopted in the present invention can not only effectively increase the content of Polygonatum cyrtonema protein peptide and Polygonatum odoratum protein peptide, but also has good stability and reproducibility.

[0193] Test Example 2 Skin irritation test

[0194] The skin irritation test was performed on the Chinese medicine protein peptide compositions involved in Examples 1 to 5, and the method and results are as follows:

[0195] 1. Test method basis: In accordance with the "Skin Irritation Test Method" in the "Technical Specifications for Safety of Cosmetics (2015 Edition)".

[0196] 2. Experimental principle: Apply the test substance once to the skin of the test animal, observe the degree of local irritation to the animal skin at a specified time interval and score it. Use self-control to evaluate the irritation of the test substance to the skin.

[0197] 3. Test samples and animals: The test substance is a 1% aqueous solution of the Chinese medicine protein peptide composition involved in Examples 1 to 5. The test animal is a common New Zealand white rabbit purchased from Guangdong Medical Experimental Animal Center.

[0198] 4. Experimental steps:

[0199] 4.1 25 white New Zealand rabbits were selected and randomly divided into 5 groups, 5 rabbits in each group, which were respectively Groups 1 to 5 of Examples.

[0200] 4.2 About 24 hours before the test, trim the hair on both sides of the spine of the New Zealand rabbit's back without damaging the epidermis. The hair removal area is about 3cm×3cm on the left and right sides.

[0201] 4.3 Take about 0.5 mL of the test substance and apply it directly to the skin, then cover it with two layers of gauze (2.5 cm × 2.5 cm) and a layer of cellophane, and then fix it with non-irritating adhesive tape and bandage. Use the other side of the skin as a control. Use a closed test with a duration of 4 hours. After the test, use warm water to remove the residual test substance.

[0202] 4.4 Observe the skin reaction at the application site 1, 24, 48 and 72 hours after the removal of the test substance, and score the skin reaction according to Table 2. The comprehensive evaluation adopts the average score of the test animals, and the skin irritation intensity is determined according to Table 3 based on the average of the highest scores at each observation time point of 24, 48 and 72 hours.

[0203] Table 2 Skin irritation reaction scores

[0204]

[0205] Table 3 Skin irritation intensity classification

[0206]

[0207] 5. Results and Analysis

[0208] Within 1 to 72 hours after administration of the Chinese medicinal protein peptide compositions of the embodiments of the present invention, no erythema or edema was observed on the skin of the New Zealand rabbits. According to the skin irritation intensity classification in Table 2, it is shown that the Chinese medicinal protein peptide compositions of the embodiments of the present invention are non-irritating to the skin.

[0209] Test Example 3 Whitening Efficacy Test

[0210] In vitro whitening efficacy tests were performed on Examples 1 to 5 and Comparative Examples 1 to 12.

[0211] 1. Methodology

[0212] The test is carried out in accordance with "T / GDCA 006-2021 Test Method for Inhibition of Tyrosinase Activity by Cosmetic Raw Materials (In Vitro Method)".

[0213] 2. Test Principle

[0214] The production process of melanin is finely regulated by tyrosinase. Tyrosinase can catalyze the conversion of L-tyrosine into DOPA, and further oxidize DOPA into DOPAquinone. This series of reactions ultimately leads to the synthesis of melanin. Therefore, tyrosinase plays a vital role in the production of melanin. It controls the production process of melanin, and its activity level is directly related to the deposition of pigment in the skin. Inhibiting the activity of tyrosinase is of great significance in delaying the browning process of the skin and reducing the production of melanin, and is one of the effective ways to whiten the skin.

[0215] Those test substances that inhibit tyrosinase activity can effectively slow down the rate at which tyrosinase catalyzes the conversion of L-tyrosine to dopaquinone. By measuring the absorbance value of dopaquinone at a wavelength of 475nm, the inhibitory effect of the test substance on tyrosinase activity can be quantitatively evaluated. The ability of a substance to inhibit tyrosinase activity can, to a certain extent, be used as an indicator to measure its whitening potential. Specifically, the higher the degree of inhibition of tyrosinase activity, the more significant the whitening effect of the substance.

[0216] 3. Materials and Reagents

[0217] The Chinese medicine protein peptide compositions were prepared according to the methods of Examples 1 to 5 and Comparative Examples 1 to 12, and dried and stored for later use.

[0218] L-Tyrosine, purchased from Macklin, purity ≥ 99.0%, biochemical reagent BR. Mushroom tyrosinase, purchased from Macklin, specification 25KU. Kojic acid, purchased from Macklin, purity ≥ 99.0%, biochemical reagent BR. The remaining reagents were of analytical grade. Thermo scientific 1530 full wavelength microplate reader.

[0219] 4. Experimental steps:

[0220] 4.1 Solution preparation

[0221] Phosphate buffer solution (PBS), pH=6.8, 0.1 mol / L: Weigh 1.361 g of potassium dihydrogen phosphate and add distilled water to a total volume of 100 mL to obtain solution A. Weigh 1.78 g of disodium hydrogen phosphate and add distilled water to 100 mL to obtain solution B. Then, mix solution A and solution B in a ratio of 1:1 to obtain a phosphate buffer solution (PBS).

[0222] L-tyrosine solution: Weigh 0.025 g of L-tyrosine, dissolve it in the PBS buffer prepared above, and perform ultrasonic treatment to promote dissolution, and finally adjust the volume to 50 mL. This solution must be prepared and used immediately to ensure its freshness and effectiveness.

[0223] Tyrosinase solution: Use PBS buffer as solvent to prepare mushroom tyrosinase to 500U / mL. After preparation, divide the solution into portions and store it at -20 degrees Celsius. At the same time, be careful to avoid secondary freezing and thawing of the solution to ensure the activity of the enzyme.

[0224] Positive control kojic acid: PBS buffer was used to dilute kojic acid into a series of mass concentration gradient solutions, with specific concentrations of 0.080 g / L, 0.040 g / L, 0.020 g / L, 0.010 g / L, 0.008 g / L, 0.005 g / L, 0.002 g / L, and 0.001 g / L.

[0225] Traditional Chinese medicine protein peptide composition solution: The traditional Chinese medicine protein peptide compositions in Examples 1 to 5 and Comparative Examples 1 to 12 were prepared into solutions with a concentration of 1.0 mg / mL using PBS buffer for later use.

[0226] Distilled water was used as blank control.

[0227] 4.2 Experimental Grouping

[0228] On the 96-well ELISA plate, the following four wells are set: solvent background well (marked as Ta), solvent reaction well (marked as Tb), sample background well (marked as Tc) and sample reaction well (marked as Td). Among them, the Ta group is the solvent background group, and its operation is not to add the substrate L-tyrosine solution, and no sample solution is added at the same time; the Tb group is the solvent reaction group, and the substrate L-tyrosine solution needs to be added, but the sample solution is not added; the Tc group is set as the sample background group, and its operation is not to add the substrate L-tyrosine solution, but the sample solution is added; and the Td group is the sample reaction group, and the substrate L-tyrosine solution and the sample solution need to be added at the same time. Three replicate wells are set for each of the above experiments to ensure the accuracy and reliability of the data.

[0229] 4.3 Test steps

[0230] According to the reagent addition amount provided in Table 4, L-tyrosine solution, sample solution / solvent and PBS buffer were added to each well in turn. After the addition is completed, the contents of each well are fully mixed and placed in a constant temperature environment of 37°C for incubation for 10 minutes. Subsequently, 20 μL of tyrosinase solution was added to each well, and the mixing reaction was continued at 37°C to ensure that the reaction time was 5min±5s. After the reaction was completed, the ELISA plate was immediately placed in the ELISA instrument to measure the absorbance values ​​of different samples and positive controls at a wavelength of 475nm. Three plate wells were set for each group for measurement, and the average value was taken. From the addition of the tyrosinase solution to the final measurement of the absorbance, the operation time of all wells remained consistent, all of which were 5min±5s.

[0231] Table 4 Tyrosinase activity inhibition test sample addition table

[0232]

[0233] Calculation of tyrosinase activity inhibition rate:

[0234] Suppression rate = [1-(Td-Tc) / (Tb-Ta)] × 100%

[0235] Where:

[0236] Td—absorbance of sample reaction well;

[0237] Tc—absorbance of sample background hole;

[0238] Tb—average absorbance of solvent reaction wells;

[0239] Ta—average absorbance of solvent background holes.

[0240] The results are shown in Table 5.

[0241] Table 5 Tyrosinase activity inhibition rate

[0242]

[0243]

[0244] It can be seen from the data in Table 5 that the Chinese medicine protein peptide composition of Examples 1 to 5 has a good inhibitory effect on tyrosinase activity, and the inhibition rate is between 59.89% and 65.47%, showing a strong whitening effect. In contrast, the inhibition rates of Comparative Examples 1 to 12 are lower, ranging from 20.87% to 56.24%, indicating that there are significant differences in the inhibitory effects between different compositions.

[0245] The inhibition rate of the positive control group (kojic acid) was 72.09%, which was higher than all the embodiments and comparative examples, indicating that the positive control had the strongest tyrosinase inhibition effect. The inhibition rate of the blank control group was only 0.21%, which showed the effectiveness of the experimental design and further verified the reliability of the experimental results. Although kojic acid has the strongest tyrosinase inhibition activity and is a special effect ingredient in whitening and freckle removal cosmetics, excessive kojic acid dosage can cause white spots on human skin and irritation. The Chinese medicine protein peptide composition is a mild and high tyrosinase inhibition activity, which is more suitable as an effective ingredient for skin care cosmetics.

[0246] Further analysis revealed that, since the number of enzymatic hydrolysis times in the extraction process of Comparative Examples 1 to 6 was different from that in the embodiment, the obtained protein peptide content was relatively low, and the tyrosinase activity inhibition rate was also low. This indicates that the tyrosinase activity inhibition rate may have a certain correlation with the protein peptide content. In other words, the level of protein peptide content may directly affect the inhibitory effect of the composition on tyrosinase.

[0247] In addition, by comparing the Chinese medicine protein peptide compositions in Examples 1 to 5 with Comparative Examples 7 to 12, it can be found that different ratios of yellow protamine peptide and yuzhu protein peptide have a significant effect on the whitening effect. When the ratio of yellow protamine peptide to yuzhu protein peptide is within a certain range (yellow protamine peptide: yuzhu protein peptide is 1 to 3: 1), the whitening effect of the composition is more significant. However, Comparative Examples 11 and 12 contain only yellow protamine peptide or yuzhu protein peptide, and their tyrosinase activity inhibition rates are relatively low. This indicates that there may be a synergistic effect between the two protein peptides, which jointly enhances the whitening effect of the composition. When only one of the protein peptides is contained at the same dosage, the whitening effect is reduced. In summary, when preparing a Chinese medicine protein peptide composition, it is crucial to reasonably control the ratio of yellow protamine peptide to yuzhu protein peptide.

[0248] Test Example 4 In vitro anti-aging efficacy test

[0249] Anti-aging efficacy test of Examples 1 to 5 and Comparative Examples 1 to 12.

[0250] 1. Methodology

[0251] The test was conducted in accordance with the General Principles of Anti-Aging-Antioxidant Evaluation Methods (T / ZGKSL001-2020).

[0252] 2. Test Principle

[0253] Excessive production of free radicals is the main cause of natural skin aging and photoaging. With the increase of age, the activity of enzymes that naturally scavenge free radicals in the skin decreases, and excess free radicals can damage skin cells, leading to the appearance of aging symptoms. Therefore, scavenging free radicals has become an effective means to delay skin aging. Cosmetics (or raw materials) with the ability to scavenge free radicals can protect skin cells from damage by reducing the generation of free radicals or removing existing free radicals, thereby delaying the skin aging process and reducing the generation of wrinkles. When evaluating the anti-aging effect of cosmetics, the scavenging ability of samples for free radicals (such as DPPH free radicals, ABTS free radicals, superoxide anions, hydroxyl free radicals, etc.) can be measured by in vitro experiments, and its anti-aging effect can be objectively evaluated. This method has the advantages of simple operation and clear results, and is one of the commonly used in vitro methods for evaluating the anti-aging effect of cosmetics.

[0254] 3. Materials and Reagents

[0255] The Chinese medicine protein peptide composition was prepared according to the methods of Examples 1 to 5 and Comparative Examples 1 to 12, and was prepared into a 0.1% aqueous solution with distilled water as a test sample. Anhydrous ethanol (Guangzhou Chemical Reagent Factory); sodium dihydrogen phosphate dihydrate (McLean); PBS phosphate buffer dry powder (White Shark, 1x); 1,1-diphenyl-2-picrylhydrazine (Solebo Technology Co., Ltd.); Trolox (McLean); total antioxidant capacity detection kit ABTS method (Biyuntian Biotechnology Co., Ltd.); Thermoscientific 1530 full-wavelength microplate reader.

[0256] Weigh 3 mg of 1,1-diphenyl-2-trinitrophenylhydrazine into a 25 mL volumetric flask and dilute to volume with 95% ethanol to prepare a 0.12 mg / mL DPPH ethanol solution.

[0257] The Chinese medicine protein peptide compositions were prepared according to the methods of Examples 1 to 5 and Comparative Examples 1 to 12, and were respectively prepared with distilled water to form 1.0 mg / mL aqueous solutions as test samples.

[0258] Accurately weigh 25.0 mg of Trolox reference substance, dilute to 25 mL with anhydrous ethanol, and refrigerate in a refrigerator at low temperature and away from light as a positive control solution.

[0259] Preparation of PBS buffer (pH = 7.4, 0.01 mol / L): weigh 8.0 g NaCl, 0.2 g KCl, 2.88 g Na2HPO4·2H2O and 0.27 g KH2PO4, dissolve in about 800 mL purified water, stir with a magnetic stirrer until completely dissolved. Adjust pH to 7.4 with 0.1 M HCl, and dilute to 1000 mL with distilled water.

[0260] 4 Experimental steps

[0261] 4.1 DPPH free radical scavenging ability determination

[0262] Set up sample wells, sample background wells, blank wells and solvent background wells in a 96-well plate. Add 30μL of the test sample solution, 150μL of distilled water, and 60μL of DPPH ethanol solution to the sample wells; replace the DPPH ethanol solution with 95% ethanol in the sample background wells. Add 180μL of distilled water and 60μL of DPPH ethanol solution to the blank wells; replace the DPPH ethanol solution with 95% ethanol in the solvent background wells. Mix thoroughly, keep away from light for 5 minutes, and measure the absorbance of each well at a wavelength of 517nm. Trolox is used as a positive control.

[0263] The calculation formula of DPPH free radical scavenging rate S1 is:

[0264] T = sample well absorbance; T0 = sample background well absorbance; C = solvent well absorbance; C0 = solvent background well absorbance

[0265] The measurement results are shown in Table 6.

[0266] Table 6 DPPH free radical scavenging rate

[0267]

[0268]

[0269] 4.2 Determination of ABTS free radical scavenging ability

[0270] According to the kit instructions, mix the ABTS solution and the ABTS working stock solution in equal proportions and store in the dark for 16 hours. When using, dilute the ABTS working stock solution with PBS buffer to make the ABTS working solution. The absorbance of the ABTS working solution minus the corresponding PBS buffer blank control should be A 734 is 0.7±0.05, corresponding to A 405 Around 1.4.

[0271] Set up sample wells, sample background wells, and blank wells in a 96-well plate. Add 10 μL of the sample solution and 200 μL of ABTS working solution to the sample wells; replace the sample solution with the corresponding solvent in the sample background wells. Add 10 μL of the corresponding solvent and 200 μL of PBS buffer (80% ethanol) to the blank wells. Mix thoroughly, store in dark for 4 minutes, and measure the A 734 The absorbance of each well was measured. Trolox was used as a positive control.

[0272] The calculation formula of ABTS free radical scavenging rate S2 is:

[0273] P = sample well absorbance; P0 = sample background well absorbance; L = solvent well absorbance

[0274] The measurement results are shown in Table 7.

[0275] Table 7 ABTS free radical scavenging rate

[0276]

[0277] It can be seen from Tables 6 and 7 that the DPPH free radical scavenging rates of the Chinese medicine protein peptide compositions of Examples 1 to 5 are between 80.68% and 83.75%, and the ABTS free radical scavenging rates are between 78.53% and 84.25%, showing strong anti-aging ability. Among them, the DPPH free radical scavenging rate of the examples is similar to the DPPH free radical scavenging rate of the positive control of 86.12%, while the ABTS free radical scavenging rate of the examples is much higher than the ABTS free radical scavenging rate of the positive control of 68.48%.

[0278] In contrast, the DPPH radical scavenging rate and ABTS radical scavenging rate of the Chinese medicine protein peptide compositions of Comparative Examples 1 to 12 are significantly lower, wherein the DPPH radical scavenging rate is between 38.21% and 72.29%, and the ABTS radical scavenging rate is between 26.97% and 75.11%. This result shows that the Polygonatum protein peptide and Polygonatum protein peptide in the Chinese medicine protein peptide composition can synergistically enhance the anti-aging effect, and their ratio and preparation process also have a significant effect on their anti-aging effect (free radical scavenging rate). Although the composition of the comparative example also shows a certain free radical scavenging ability, the overall effect is not as good as the composition of the embodiment.

[0279] In summary, the Chinese medicine protein peptide compositions of Examples 1 to 5 have strong in vitro anti-aging effects and outstanding antioxidant capacity, and are expected to provide an effective solution for delaying skin aging. Although the compositions of the comparative examples also have certain anti-aging effects, their overall performance is not as good as that of the compositions of the examples, which further illustrates the importance of the raw material ratio and preparation process of the Chinese medicine protein peptide compositions.

[0280] Test Example 5 Moisturizing Efficacy Test

[0281] Moisturizing efficacy test of Examples 1 to 5 and Comparative Examples 1 to 12.

[0282] 1. Method basis: The test was conducted in accordance with “QBT4256-2011 Guidelines for Evaluation of Moisturizing Efficacy of Cosmetics”.

[0283] 2. Materials and reagents: The Chinese medicine protein peptide composition was prepared according to the methods of Examples 1 to 5 and Comparative Examples 1 to 12, and was prepared into 0.1% aqueous solution with distilled water as the test sample. The reference sample was 1% glycerol, and the blank control was 100% distilled water.

[0284] 3. Experimental method: 95 healthy subjects aged 18 to 55 were selected on a voluntary basis. They were required to have no history of serious diseases, immunodeficiency, or allergies, and had not used hormones or immunosuppressants in the past month, and had not participated in other experiments. The subjects were randomly divided into 19 groups, 5 people in each group, namely, Example 1 to 5 groups, Comparative Example 1 to 12 groups, a control group, and a blank group. During the experiment, the subjects did not apply any drugs or cosmetics unrelated to the experiment.

[0285] Before the test, clean the inner forearms of the subjects' hands and wipe them with dry tissues, and mark the measurement area on the inner forearms of the subjects' hands. Before the formal test, sit quietly in a standard room (test environment temperature: (22±1)℃, humidity: (50±5)%, real-time dynamic monitoring) for at least 30 minutes, do not drink water during this period, expose your forearms, place them in the test state, and keep relaxed.

[0286] During the experiment, a 3×3 cm area was marked on the inside of the left and right arms. 2 Test area, multiple areas can be marked on the same arm, and the interval between each area is 1 cm. The test samples and control samples are randomly assigned to the left and right arms. The Corneometer CM 825 skin moisture tester is used to measure and record the skin moisture content of the test area. The results are expressed as moisture measurement value (Moisture Measurement Value, MMV) in units of CU (Corneometer Units). Each area is measured in parallel 5 times, and the average value is taken as the final test value. First, the initial (blank) value of each test area is measured, and then the value is calculated according to (2.0±0.1) mg / cm 2 The test sample was evenly applied to the designated area using a latex fingertip at a dosage of 1.5 mg / ml. The skin moisture content of the test area and the control area was measured 1 hour and 2 hours after application. To ensure the consistency of the test, all tests on the same volunteer were completed by the same measurement personnel. The skin moisture content growth rate at 2 hours was calculated according to the following formula, and the average value was taken. The higher the skin moisture content growth rate, the better the moisturizing effect. The test results are shown in Table 8 below.

[0287] Moisture content growth rate = (MMVt-MMV0) / MMV0×100%

[0288] Where MMV0 is the skin MMV before use; MMVt is the skin MMV at time period t after use.

[0289] Table 8 Skin moisture content and 2h moisture content growth rate

[0290]

[0291] It can be seen from the data in Table 8 that after the Chinese medicine protein peptide composition in the embodiment is applied to the skin of the subject, the skin moisture content growth rate within 2 hours is between 45.60% and 49.30%. This fully demonstrates that the Chinese medicine protein peptide composition in the embodiment has a strong moisturizing effect, which is similar to the moisturizing effect of the positive control group. In contrast, the moisturizing effect of the Chinese medicine protein peptide composition in Comparative Examples 1 to 12 is significantly weaker. The 2-hour skin moisture content growth rate of these compositions ranges from only 16.32% to 43.82%, which is much lower than the composition of the embodiment. This confirms that there are significant differences in moisturizing effects between different Chinese medicine protein peptide compositions.

[0292] The weak moisturizing ability of comparative examples 1 to 6 is related to the number of enzymatic hydrolysis times in their extraction process. Since the number of enzymatic hydrolysis times of these two groups of comparative examples is different from that of the examples, the obtained protein peptide content is relatively low. At the same time, the growth rate of the skin moisture content in 2 hours is also correspondingly low. This shows that different enzymatic hydrolysis times and the resulting differences in protein peptide content may have an important impact on the moisturizing ability of protein peptides.

[0293] By comparing the data of the examples with those of comparative examples 11 to 12, it can be found that the moisturizing ability of the Chinese medicine protein peptide composition also has a synergistic effect. Comparative examples 11 and 12 contain only yellow spherical protein peptide or yuzhu protein peptide, respectively, and their 2-hour skin moisture content growth rate is relatively low. This shows that when yellow spherical protein peptide and yuzhu protein peptide are used together, their moisturizing effect is significantly better than the effect of using either protein peptide alone.

[0294] By comparing the data of the embodiment with the comparative examples 7 to 10, it can be seen that the different ratios of Polygonatum protein peptide and Polygonatum protein peptide in the traditional Chinese medicine protein peptide composition have a significant effect on the moisturizing ability. When the ratio of the two protein peptides is within a certain range (the ratio in the embodiment), the moisturizing effect of the traditional Chinese medicine protein peptide composition is more significant.

[0295] Therefore, this test example shows that under the preparation conditions and raw material dosage ratio described in the present invention, a Chinese medicine protein peptide composition with significant moisturizing effect can be obtained.

Claims

1. A Chinese medicine protein peptide composition, characterized in that: The invention is composed of yellow protamine peptide and polygonatum protein peptide in a mass ratio of 1 to 3:1; the preparation method of the yellow protamine peptide and polygonatum protein peptide is that a composite enzyme is used for enzymatic hydrolysis to obtain crude protein, and the crude protein is then enzymatically hydrolyzed twice with a composite alkaline protein and pepsin in sequence to obtain small molecule peptides with a molecular weight below 10KDa, the composite enzyme is α-mannosidase, β-galactosidase, α-glucosidase and β-glucosidase, and the composite alkaline protein is trypsin and papain.

2. The Chinese medicine protein peptide composition according to claim 1, characterized in that: The preparation method of the polyanthocyanin peptide and the polyanthocyanin peptide comprises the following steps: S1. Performing hydrothermal extraction on the Chinese medicine raw materials to obtain a Chinese medicine extract; S2. The Chinese medicine extract is initially hydrolyzed using a composite enzyme to obtain crude protein of Chinese medicine; the composite enzyme is α-mannosidase, β-galactosidase, α-glucosidase and β-glucosidase; S3. The crude protein solution of traditional Chinese medicine is hydrolyzed for a second time using a composite alkaline protease to obtain a second hydrolyzate; the composite alkaline protease is trypsin and papain; S4. performing a third enzymatic hydrolysis on the second enzymatic hydrolyzate using pepsin to obtain a third enzymatic hydrolyzate; S5. The third enzymatic hydrolysate was ultrafiltered and post-treated using a membrane with a molecular weight cutoff of 10 KDa to obtain a Chinese medicine protein peptide; The Chinese medicinal raw material is Polygonatum odoratum or Polygonatum sibiricum.

3. The Chinese medicine protein peptide composition according to claim 2, characterized in that: The mass ratio of α-mannosidase, β-galactosidase, α-glucosidase and β-glucosidase in the complex enzyme is 0.8-1.2: 0.8-1.2: 0.8-1.2: 0.8-1.

2.

4. The Chinese medicine protein peptide composition according to claim 2, characterized in that: The mass ratio of trypsin to papain in the composite alkaline protease is 0.8-1.2:0.8-1.

2.

5. The Chinese medicine protein peptide composition according to claim 2, characterized in that: When the Chinese medicine raw material is Polygonatum sibiricum, the amount of the composite enzyme in step S2 is 1-3% of the mass of the Chinese medicine extract; the amount of the composite alkaline protease in step S3 is 1-5% of the mass of the Chinese medicine crude protein solution; the amount of pepsin in step S4 is 1-5% of the mass of the second enzymatic hydrolyzate.

6. The Chinese medicine protein peptide composition according to claim 2, characterized in that: When the Chinese medicine raw material is Polygonatum odoratum, the amount of the complex enzyme in step S2 is 1.5-3.5% of the mass of the Chinese medicine extract; the amount of the complex alkaline protease in step S3 is 1.5-5.5% of the mass of the Chinese medicine crude protein solution; the amount of pepsin in step S4 is 1.5-5.5% of the mass of the second enzymatic hydrolyzate.

7. Use of the Chinese medicinal protein peptide composition according to any one of claims 1 to 6 in the preparation of whitening, anti-aging and moisturizing cosmetics.

8. The use according to claim 7, wherein the cosmetic is an aqueous solution, emulsion, cream or essence.

9. A cosmetic, characterized in that: A Chinese medicinal protein peptide composition comprising any one of claims 1 to 6.

10. The cosmetic according to claim 9, characterized in that: The mass percentage of the traditional Chinese medicine protein peptide composition in cosmetics is 0.1-5%.

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