Multi-Anti-oxidization composition containing recombinant collagen, preparation method therefor, and use thereof

Abstract

The present application provides a multi-anti-oxidation composition containing recombinant collagen, a preparation method therefor, and use thereof. The multi-anti-oxidation composition containing the recombinant collagen comprises the recombinant collagen and an anti-oxidation embedding solution. The anti-oxidation embedding solution comprises hydroxytyrosol, ergothioneine, and astaxanthin. In the multi-anti-oxidation composition containing the recombinant collagen of the present application, on the basis of the recombinant collagen, the three anti-oxidation components of hydroxytyrosol, ergothioneine, and astaxanthin are added, and since the stability of the anti-oxidation components is poor, the three anti-oxidation components are subjected to multi-layer embedding treatment of schizophyllan-pectin in the present application, such that the composition achieves a stable multi-anti-oxidation effect, thereby improving the stability of the active ingredients, and achieving stable exertion of the active ingredients.

Description

A multi-antioxidant composition containing recombinant collagen and its preparation method and application Technical Field

[0001] The present application belongs to the field of cosmetic technology, and specifically relates to a multi-antioxidant composition containing recombinant collagen, and a preparation method and application thereof. Background Art

[0002] As we age, our body functions decline, the skin's epidermis becomes dry and thinner, collagen and elastin gradually disappear, and the fine mesh structure formed by the interweaving of collagen and elastic fibers is damaged, resulting in the appearance of wrinkles of varying sizes. Furthermore, the accumulation of oxygen free radicals is also a major cause of skin aging. Therefore, antioxidants are of great significance in preventing skin aging.

[0003] Collagen has the ability to stretch, maintain skin elasticity and promote cell growth. When used in skin care products, it can have moisturizing, repairing, and anti-wrinkle effects. It also has good biodegradability and bio-cosmetic properties. Therefore, collagen is also beginning to be used more and more in skin care products.

[0004] Hydroxytyrosol is a natural polyphenolic compound with diverse biological and pharmacological activities that can be extracted from olive oil. It plays a role in cancer prevention and treatment, anti-inflammatory activities, the prevention and treatment of cardiovascular and cerebrovascular diseases, the prevention and treatment of coronary heart disease, and the protection of retinal pigment epithelial cells. Its molecular structure not only possesses phenolic hydroxyl groups like other phenolic substances, but also contains alcoholic hydroxyl groups on the ethanol chain attached to the benzene ring. This gives hydroxytyrosol excellent antioxidant activity, effectively removing exogenous and endogenous oxidants and free radicals by controlling the oxidation of many unsaturated fatty acids.

[0005] Ergothioneine is a natural antioxidant and an important physiologically active substance in the body. It has multiple functions such as antioxidant, anti-aging, and anti-inflammatory, and can be used in food, biomedicine, cosmetics and other fields.

[0006] Astaxanthin is a chain-breaking antioxidant with strong antioxidant capacity. It can provide electrons to react with free radicals, thereby achieving the purpose of scavenging free radicals (antioxidation). Studies have shown that astaxanthin's free radical scavenging ability is stronger than many other antioxidant ingredients.

[0007] CN113966838A discloses an astaxanthin nanostructured lipid carrier—chitosan gel microparticles and a preparation method. The microparticles comprise an astaxanthin nanostructured lipid carrier and chitosan coated on the surface of the astaxanthin nanostructured lipid carrier. This method improves the stability and bioavailability of astaxanthin and enhances its water dispersibility. However, the cost of the excipients and equipment required to prepare the liposomes is relatively high. Furthermore, the production process for chitosan, another key ingredient, is highly environmentally unfriendly, requiring high-temperature treatment with strong acids and bases, generating large amounts of wastewater, and consuming significant amounts of energy.

[0008] CN112156036A discloses active ingredient particles based on coating technology and a preparation method thereof, relating to the technical field of cosmetics. The raw material components include astaxanthin, niacinamide, fruit acid, ergothioneine, and jojoba esters. Using jojoba esters as a coating, the four active ingredients (astaxanthin, niacinamide, fruit acid, and ergothioneine) are combined to form active ingredient particles. This effectively solves the contradiction that niacinamide and fruit acid cannot be used simultaneously, while also preventing product discoloration and failure caused by ergothioneine and astaxanthin.

[0009] The three main active ingredients, hydroxytyrosol, ergothioneine, and astaxanthin, all have the problems of instability and easy decomposition in practical applications. Among them, hydroxytyrosol, like many polyphenol molecules, is extremely sensitive to temperature and acidity and is prone to discoloration, which greatly hinders its use in cosmetics. Ergothioneine is highly water-soluble but insoluble in oil, and its skin permeability is poor, resulting in low bioavailability. In addition, in practical applications, ergothioneine is easy to discolor and has the disadvantages of being easily decomposed by light and having poor stability, which will cause the degradation of the effective ingredients of ergothioneine and greatly reduce its efficacy. Although natural astaxanthin has strong antioxidant properties, it has a highly unsaturated structure, which makes it tend to chemically degrade when exposed to conditions such as high temperature and light, which promotes its fading and the decline of its biological activity, while limiting its application in the fields of food, medicine, and cosmetics.

[0010] Therefore, developing an antioxidant composition that can achieve multiple antioxidant effects while improving the stability of active ingredients is a research focus in this field.

[0011] Summary of the Invention

[0012] The following is a summary of the subject matter described in detail herein. This summary is not intended to limit the scope of the claims.

[0013] In view of the shortcomings of the existing technology, the purpose of this application is to provide a multi-antioxidant composition containing recombinant collagen and its preparation method and application, which can achieve multiple antioxidant effects while improving the stability of active ingredients.

[0014] To achieve this goal, this application adopts the following technical solutions:

[0015] In a first aspect, the present application provides a multiple antioxidant composition containing recombinant collagen, wherein the multiple antioxidant composition containing recombinant collagen comprises recombinant collagen and an antioxidant embedding solution;

[0016] The antioxidant embedding solution comprises hydroxytyrosol, ergothioneine and astaxanthin.

[0017] The multi-antioxidant composition containing recombinant collagen of the present application is based on recombinant collagen and adds three antioxidant ingredients, namely hydroxytyrosol, ergothioneine and astaxanthin. By encapsulating the three antioxidant ingredients, stable multi-antioxidant effects are achieved, the stability of the active ingredients is improved, and the stable performance of the active ingredients is achieved.

[0018] In one embodiment, the multiple antioxidant composition containing recombinant collagen includes, in parts by weight: 0.1-0.5 parts of recombinant collagen (for example, 0.2 parts, 0.3 parts, 0.4 parts, etc.), and 1.5-10 parts of antioxidant embedding solution (for example, 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, etc.).

[0019] In one embodiment, the mass ratio of hydroxytyrosol, ergothioneine and astaxanthin in the antioxidant embedding solution is (0.1-5):(1-5):(0.05-3), for example, it can be 0.5:1:1, 1:1:1, 2:1:1, 3:1:1, 4:1:1, 0.5:2:1, 1:2:1, 2:2:1, 3:2:1, 4:2:1, 0.5:3:1, 1:3:1, 2:3:1, 3:3:1, 4:3:1, 0.5:4:1, 1:4:1, 2:4:1, 3:4:1, 4:4:1, 0.5:1:2, 1:1:2, 2:1:2, 3:1:2, 4:1:2, etc.

[0020] In one embodiment, the mass ratio of hydroxytyrosol, ergothioneine and astaxanthin in the antioxidant embedding solution is (1-3):(3-5):(0.1-2).

[0021] In one embodiment, the antioxidant embedding solution is prepared by the following method, which comprises:

[0022] (1) mixing hydroxytyrosol, ergothioneine and astaxanthin with linseed oil, heating and ultrasonically dispersing to obtain an oil phase; dissolving the Quillaja bark extract in a phosphate buffer to obtain an aqueous phase; and mixing the oil phase and the aqueous phase to obtain a primary emulsion;

[0023] (2) uniformly mixing the primary emulsion with a schizophyllan solution and a phosphate buffer to obtain a secondary emulsion;

[0024] (3) Evenly mixing the secondary emulsion, pectin solution, and phosphate buffer to obtain the antioxidant embedding solution.

[0025] In one embodiment, the heating temperature is 45-55°C, for example, 46°C, 48°C, 50°C, 52°C, 54°C, etc., and the heating time is 1.5-2.5h, for example, 1.6h, 1.8h, 2h, 2.2h, 2.4h, etc.

[0026] In one embodiment, the ultrasonic treatment time is 2-5 min, for example, 2.5 min, 3 min, 4 min, etc.

[0027] In one embodiment, the mass ratio of the oil phase to the water phase in the primary emulsion is 1:(8-12), for example, 1:9, 1:10, 1:11, etc.

[0028] In one embodiment, the mass percentage of the Quillaja bark extract in the aqueous phase is 0.8-1.2%, for example, 0.9%, 1%, 1.1%, etc.

[0029] In one embodiment, the mass percentage of the total amount of hydroxytyrosol, ergothioneine and astaxanthin in the oil phase is 10-20%, for example, 12%, 14%, 16%, 18%, etc.

[0030] In one embodiment, the mass percentage of Schizophyllan in the Schizophyllan solution is 0.8-1.2%, for example, 0.9%, 1%, 1.1%, etc.

[0031] In one embodiment, the solvent of the Schizophyllan solution is an acetic acid aqueous solution, and the mass percentage of acetic acid in the acetic acid aqueous solution is 0.8-1.2%, for example, 0.9%, 1%, 1.1%, etc.

[0032] In one embodiment, the mass percentage of pectin in the pectin solution is 3-5%, for example, 3.5%, 4%, 4.5%, etc.

[0033] In one embodiment, the solvent of the pectin solution is phosphate buffer.

[0034] The numerical range described in this application includes not only the point values ​​listed above, but also any point values ​​between the above numerical ranges that are not listed. Due to limited space and for the sake of simplicity, this application no longer exhaustively lists the specific point values ​​included in the range.

[0035] In a second aspect, the present application provides a method for preparing the multi-antioxidant composition containing recombinant collagen as described in the first aspect, the preparation method comprising: uniformly mixing the recombinant collagen and the antioxidant embedding solution to obtain the composition.

[0036] In a third aspect, the present application provides a use of the multi-antioxidant composition containing recombinant collagen as described in the first aspect in cosmetics.

[0037] In a fourth aspect, the present application provides a multi-antioxidant moisturizing lotion, the raw materials of which include: the multi-antioxidant composition containing recombinant collagen described in the first aspect, an emollient, a moisturizer, an emulsifier, and a thickener.

[0038] Compared with the prior art, this application has the following beneficial effects:

[0039] 1. The multiple antioxidant composition containing recombinant collagen of the present application is based on recombinant collagen and adds three antioxidant ingredients, hydroxytyrosol, ergothioneine and astaxanthin. Since the antioxidant ingredients have poor stability, the present application performs a multi-layer embedding treatment of the three antioxidant ingredients with Schizophyllan-pectin to enable the composition to achieve stable multiple antioxidant effects, improve the stability of the active ingredients, and achieve stable performance of the active ingredients.

[0040] 2. The antioxidant embedding solution of the present application is prepared by means of synthetic biology, with milder reaction conditions, simpler process, less carbon emissions during the reaction process, and the embedding treatment process is more low-carbon and environmentally friendly than the existing technology.

[0041] Still other aspects will become apparent upon reading and understanding the detailed description. DETAILED DESCRIPTION

[0042] The technical solution of the present application is further described below through specific implementation methods. Those skilled in the art should understand that the embodiments are only used to help understand the present application and should not be regarded as specific limitations of the present application.

[0043] As used herein, the terms "comprises," "including," "having," "containing" or any other variations thereof, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that comprises a listed element is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.

[0044] "Optional" or "either" means that the subsequently described event or circumstance can or cannot occur, and that the description includes instances where the event occurs and instances where it does not.

[0045] The indefinite articles "a" and "an" before an element or component of the present application do not limit the quantity requirement (i.e., the number of times the element or component appears). Therefore, "a" or "an" should be interpreted as including one or at least one, and elements or components in the singular also include plural forms, unless the quantity clearly refers to only the singular form.

[0046] The terms "one embodiment," "some embodiments," "exemplarily," "specific examples," or "some examples" used herein mean that a specific feature, structure, material, or characteristic described in conjunction with the embodiment or example is included in at least one embodiment or example of the present application. In this document, the schematic descriptions of the above terms do not necessarily refer to the same embodiment or example.

[0047] In this application, unless otherwise specified, percentages and contents are all by mass. Unless otherwise specified, the experimental methods used are conventional methods, and the materials and reagents used can be purchased from commercial sources.

[0048] Preparation Example 1

[0049] This preparation example provides an antioxidant embedding solution, wherein the raw materials of the antioxidant embedding solution include: 2 parts of hydroxytyrosol, 4 parts of ergothioneine, and 1 part of astaxanthin;

[0050] The preparation method is as follows:

[0051] (1) Hydroxytyrosol, ergothioneine, and astaxanthin were mixed with linseed oil, heated at 50° C. for 2 h, and then ultrasonically dispersed for 2 min to obtain an oil phase, wherein the content of the three antioxidant components in the oil phase was 15%; Q-Naturale extract was dissolved in phosphate buffer (pH 7.0) to obtain an aqueous phase, wherein the content of Q-Naturale extract in the aqueous phase was 1%; and the oil phase and the aqueous phase were mixed at 10,000 rpm to obtain a primary emulsion;

[0052] (2) the primary emulsion was mixed with a schizophyllan solution (the solvent was a 1% acetic acid aqueous solution, the concentration of schizophyllan was 1%) and a phosphate buffer (pH 4.0) in a volume ratio of 1:1:2 to obtain a secondary emulsion;

[0053] (3) The secondary emulsion was mixed with pectin solution (the solvent was phosphate buffer pH 5.0, and the pectin concentration was 4%) and phosphate buffer (pH 4.0) in a volume ratio of 2:1:1 to obtain the antioxidant embedding solution.

[0054] Preparation Example 2

[0055] This preparation example provides an antioxidant embedding solution, wherein the raw materials of the antioxidant embedding solution include: 3 parts of hydroxytyrosol, 3 parts of ergothioneine, and 2 parts of astaxanthin;

[0056] The preparation method is as follows:

[0057] (1) Hydroxytyrosol, ergothioneine, and astaxanthin were mixed with linseed oil, heated at 55° C. for 1.5 h, and then ultrasonically dispersed for 2.5 min to obtain an oil phase, wherein the content of the three antioxidant components in the oil phase was 10%; Q-Naturale extract was dissolved in phosphate buffer (pH 7.0) to obtain an aqueous phase, wherein the content of Q-Naturale extract in the aqueous phase was 0.8%; the oil phase and the aqueous phase were mixed at 10,000 rpm to obtain a primary emulsion;

[0058] (2) the primary emulsion was mixed with a schizophyllan solution (the solvent was a 0.8% acetic acid aqueous solution, the concentration of schizophyllan was 1.2%) and a phosphate buffer (pH 4.0) in a volume ratio of 1:1:2 to obtain a secondary emulsion;

[0059] (3) The secondary emulsion was mixed evenly with a pectin solution (the solvent was a phosphate buffer with a pH of 5.0 and a pectin concentration of 3%) and a phosphate buffer (pH 4.0) at a volume ratio of 2:1:1 to obtain the antioxidant embedding solution.

[0060] Preparation Example 3

[0061] This preparation example provides an antioxidant embedding solution, wherein the raw materials of the antioxidant embedding solution include: 1 part of hydroxytyrosol, 5 parts of ergothioneine, and 0.1 part of astaxanthin;

[0062] The preparation method is as follows:

[0063] (1) Hydroxytyrosol, ergothioneine, and astaxanthin were mixed with linseed oil, heated at 45° C. for 2.5 h, and then ultrasonically dispersed for 2.5 min to obtain an oil phase, wherein the content of the three antioxidant components in the oil phase was 20%; Q-Naturale extract was dissolved in phosphate buffer (pH 7.0) to obtain an aqueous phase, wherein the content of Q-Naturale extract in the aqueous phase was 1.2%; and the oil phase and the aqueous phase were mixed at 10,000 rpm to obtain a primary emulsion;

[0064] (2) the primary emulsion was mixed with a schizophyllan solution (the solvent was a 1.2% acetic acid aqueous solution, the concentration of schizophyllan was 0.8%) and a phosphate buffer (pH 4.0) in a volume ratio of 1:1:2 to obtain a secondary emulsion;

[0065] (3) The secondary emulsion was mixed evenly with a pectin solution (the solvent was a phosphate buffer with a pH of 5.0 and a pectin concentration of 5%) and a phosphate buffer (pH 4.0) in a volume ratio of 2:1:1 to obtain the antioxidant embedding solution.

[0066] Preparation Example 4

[0067] This preparation example provides an antioxidant embedding solution, the raw materials of which include: 5 parts of hydroxytyrosol, 1 part of ergothioneine, and 0.05 parts of astaxanthin; the preparation method refers to Preparation Example 1.

[0068] Preparation Example 5

[0069] This preparation example provides an antioxidant embedding solution, the raw materials of which include: 0.1 parts of hydroxytyrosol, 5 parts of ergothioneine, and 3 parts of astaxanthin; the preparation method refers to Preparation Example 1.

[0070] Comparative Preparation Example 1

[0071] This comparative preparation example provides an antioxidant embedding solution, which differs from Preparation Example 1 only in that hydroxytyrosol is not added, and the insufficient part is supplemented by ergothioneine and astaxanthin in a ratio of 4:1; the preparation method refers to Preparation Example 1.

[0072] Comparative Preparation Example 2

[0073] This comparative preparation example provides an antioxidant embedding solution, which differs from Preparation Example 1 only in that ergothioneine is not added, and the insufficient part is supplemented by hydroxytyrosol and astaxanthin in a ratio of 2:1; the preparation method refers to Preparation Example 1.

[0074] Comparative Preparation Example 3

[0075] This comparative preparation example provides an antioxidant embedding solution, which differs from Preparation Example 1 only in that astaxanthin is not added, and the insufficient part is supplemented by hydroxytyrosol and ergothioneine in a ratio of 1:2; the preparation method refers to Preparation Example 1.

[0076] Comparative Preparation Example 4

[0077] This comparative preparation example provides an antioxidant embedding solution, which differs from Preparation Example 1 only in the preparation method. Step (2) is not performed, and the primary emulsion obtained in step (1) is directly subjected to step (3). The raw materials and amounts are referred to Preparation Example 1.

[0078] Comparative Preparation Example 5

[0079] This comparative preparation example provides an antioxidant embedding solution, which differs from Preparation Example 1 only in the preparation method. Step (3) is not performed. The secondary emulsion obtained in step (2) is the antioxidant embedding solution. The raw materials and amounts are referred to Preparation Example 1.

[0080] Comparative Preparation Example 6

[0081] This comparative preparation example provides an antioxidant embedding solution, which differs from Preparation Example 1 only in the preparation method. The raw materials and amounts used refer to Preparation Example 1.

[0082] The preparation method of this comparative preparation example includes:

[0083] (1) Hydroxytyrosol, ergothioneine, and astaxanthin were mixed with linseed oil, heated at 50° C. for 2 h, and then ultrasonically dispersed for 2 min to obtain an oil phase, wherein the content of the three antioxidant components in the oil phase was 15%; Q-Naturale extract was dissolved in phosphate buffer (pH 7.0) to obtain an aqueous phase, wherein the content of Q-Naturale extract in the aqueous phase was 1%; and the oil phase and the aqueous phase were mixed at 10,000 rpm to obtain a primary emulsion;

[0084] (2) mixing the primary emulsion with a pectin solution (the solvent is a phosphate buffer with a pH of 5.0 and a pectin concentration of 4%) and a phosphate buffer (pH 4.0) in a volume ratio of 2:1:1 to obtain a secondary emulsion;

[0085] (3) The secondary emulsion was mixed with a schizophyllan solution (the solvent was a 1% acetic acid aqueous solution, and the concentration of schizophyllan was 1%) and a phosphate buffer (pH 4.0) in a volume ratio of 1:1:2 to obtain the antioxidant embedding solution.

[0086] Example 1

[0087] This embodiment provides a multiple antioxidant composition, which includes: 0.3 parts of recombinant collagen and 5 parts of the antioxidant embedding solution prepared in Preparation Example 1. The recombinant collagen and the antioxidant embedding solution are evenly mixed to obtain the composition.

[0088] Example 2

[0089] This embodiment provides a multiple antioxidant composition, which includes: 0.5 parts of recombinant collagen and 1.5 parts of the antioxidant embedding solution prepared in Preparation Example 2. The recombinant collagen and the antioxidant embedding solution are evenly mixed to obtain the composition.

[0090] Example 3

[0091] This embodiment provides a multiple antioxidant composition, which includes: 0.1 parts of recombinant collagen and 10 parts of the antioxidant embedding solution prepared in Preparation Example 3. The recombinant collagen and the antioxidant embedding solution are evenly mixed to obtain the composition.

[0092] Examples 4-5

[0093] This embodiment provides a multiple antioxidant composition, which differs from Example 1 only in that the antioxidant embedding solution prepared in Preparation Example 1 is replaced with an equal amount of antioxidant embedding solution prepared in Preparation Examples 4-5; other raw materials, amounts and preparation methods are similar to those in the examples.

[0094] Comparative Examples 1-6

[0095] This comparative example provides a multiple antioxidant composition, which differs from Example 1 only in that the antioxidant embedding solution prepared in Preparation Example 1 is replaced with an equal amount of the antioxidant embedding solution prepared in Preparation Examples 1-65; other raw materials, amounts and preparation methods refer to the examples.

[0096] Comparative Example 7

[0097] This embodiment provides a multiple antioxidant composition, which includes: 0.3 parts of recombinant collagen and 5 parts of an antioxidant compound; wherein the antioxidant compound is obtained by compounding hydroxytyrosol, ergothioneine and astaxanthin in a ratio of 1:5:0.1, and no encapsulation treatment is performed; the recombinant collagen and the antioxidant compound are evenly mixed to obtain a composite.

[0098] Test Example 1

[0099] Evaluation of DPPH free radical scavenging ability

[0100] The ability to scavenge DPPH (1,1-diphenyl-2-trinitrophenylhydrazine) free radicals reflects, to a certain extent, a substance's antioxidant capacity. A higher free radical scavenging rate indicates stronger antioxidant capacity and, therefore, stronger anti-aging properties. Therefore, the anti-aging effects of an anti-aging composition can be assessed by studying its DPPH free radical scavenging capacity.

[0101] The multiple antioxidant compositions prepared in the examples and comparative examples were diluted with DMSO to a sample with a mass fraction of 2%, and the following test was performed:

[0102] Add 2.0 mL of 45.8 mg / L DPPH test solution and the sample to be tested to the test tube, add anhydrous ethanol to the total volume to 3 mL, shake well, and react in the dark for 30 minutes. Then use a 1 cm cuvette to measure the absorbance at a wavelength of 517 nm, which is recorded as A. 10 ; Add 2mL of anhydrous ethanol and the corresponding volume of the sample to be tested to the test tube, and add anhydrous ethanol to make up the total volume to 3mL. The measured absorbance is recorded as A 11 ; Add 2mL of DPPH test solution and 1mL of anhydrous ethanol to the test tube, and record the measured absorbance as A 12 , calculate the DPPH free radical scavenging rate of the test solution (Y');

[0103] The calculation formula for DPPH free radical scavenging rate is:

[0104] Where A 10 A is the absorbance value of the system after the sample to be tested removes DPPH, 11 is the absorbance value of the system after removing DPPH from the sample blank control, A 12 is the absorbance value of the reaction system before adding the drug.

[0105] The test results are shown in Table 1.

[0106] Test Example 2

[0107] Hydroxyl free radical scavenging assay

[0108] Main reagents: salicylic acid (analytical pure, Sinopharm Chemical Reagent Co., Ltd.), ferrous sulfate (analytical pure, Sinopharm Chemical Reagent Co., Ltd.), anhydrous ethanol (analytical pure, Tianjin Fuyu Fine Chemical Co., Ltd.), and hydrogen peroxide.

[0109] Main equipment: Spectra Max M5 Molecular Device

[0110] Experimental Method: A sample group (Example and Comparative Example) containing the multi-antioxidant composition was diluted to 1 / 50, and a negative control group was prepared. 0.1 part ferrous sulfate, 0.1 part ethanol, 0.1 part salicylic acid, 0.1 part sample or negative control, 1 part deionized water, and 0.1 part hydrogen peroxide were added to a 1.5 mL centrifuge tube, capped tightly, and shaken upside down. The tube was incubated in a 37°C water bath for 15 minutes. The solution was then transferred to a 96-well plate, with triplicate wells per group. The absorbance at 510 nm was measured using a microplate reader.

[0111] The hydroxyl radical scavenging rate was calculated by the following formula:

[0112] Hydroxyl radical scavenging rate % = [A0-(A X -A X0 )] / A0×100%, where A0 is the absorbance of the negative control group, A X is the absorbance of the sample group, A X0 The test results are shown in Table 1.

[0113] Test Example 3

[0114] Skin stratum corneum moisture content test

[0115] Testing instrument: Skin stratum corneum moisture content meter milipure;

[0116] Test sample: an antioxidant moisturizing lotion containing the multiple antioxidant compositions prepared in the Examples and Comparative Examples, wherein the ingredients of the antioxidant moisturizing lotion include: 80 parts of ionized water, 4 parts of squalane, 3 parts of glycerin, 5 parts of betaine, 2 parts of sodium hyaluronate, 2 parts of sodium stearoyl lactylate, and 0.5 parts of guar gum;

[0117] Testing phase: 2 weeks (D2), 4 weeks (D4);

[0118] Test subjects: 30 healthy women aged 20-45 years;

[0119] Measurement area: The test location is near the temple on the right side of the volunteer's face;

[0120] Test parameter explanation:

[0121] Water content of the stratum corneum: The higher the value, the more water the stratum corneum contains. Unit: Comeometer Unit, abbreviated as CU. Water growth rate (%) = (D n -D0)×100% / D0, where Dn is the water content of the subject's skin at D2 and D4, and D0 is the initial water content of the subject's skin. The test results are shown in Table 1.

[0122] Table 1

[0123] According to the tabular data, it can be seen from Example 1-3 that when the mass ratio of hydroxytyrosol, thioneine and astaxanthin in the antioxidant embedding solution is (1-3): (3-5): (0.1-2), the antioxidant and moisturizing effects are the best. It can be seen from Example 1-3 and Example 4-5 that when the mass ratio of hydroxytyrosol, thioneine and astaxanthin in the antioxidant embedding solution is (0.1-5): (1-5): (0.05-3), the antioxidant and moisturizing effects are not as good as Example 1-3.

[0124] It can be seen from Example 1 and Comparative Examples 1-3 that when the raw material of the antioxidant embedding solution is a combination of hydroxytyrosol, ergothioneine and astaxanthin, the effect is best. If any one of the raw materials is missing, the antioxidant and moisturizing effects of the resulting composition will decrease.

[0125] It can be seen from Example 1 and Comparative Examples 4-5 that when the secondary embedding of the antioxidant embedding solution is retained only once, whether the Schizophyllan or pectin is retained, the antioxidant efficacy of the resulting composition will be greatly reduced; it can be seen from Example 1 and Comparative Example 6 that when the two embeddings in the embedding process are swapped in order, pectin embedding is performed first, and then Schizophyllan embedding is performed, the antioxidant efficacy of the resulting composition will also be greatly reduced; it can be seen from Example 1 and Comparative Example 7 that when the antioxidant component is not embedded and is directly combined with collagen, the antioxidant and moisturizing properties of the resulting composition are greatly reduced.

[0126] The applicant declares that while the present application uses the above-mentioned embodiments to illustrate the process of the present application, the present application is not limited to the above-mentioned embodiments, which does not mean that the present application must rely on the above-mentioned embodiments in order to be implemented. Those skilled in the art should understand that any improvements to the present application, equivalent replacements for the raw materials of the present application's products, addition of auxiliary ingredients, and selection of specific methods, etc., fall within the scope of protection and disclosure of the present application.

[0127] The preferred embodiments of the present application are described in detail above. However, the present application is not limited to the specific details of the above embodiments. Within the technical concept of the present application, various simple modifications can be made to the technical solution of the present application, and these simple modifications all fall within the scope of protection of the present application.

[0128] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any appropriate manner unless there is any contradiction. In order to avoid unnecessary repetition, this application will not further describe various possible combinations.

Claims

1. A multi-antioxidant composition containing recombinant collagen, comprising recombinant collagen and an antioxidant embedding solution; The antioxidant embedding solution comprises hydroxytyrosol, ergothioneine and astaxanthin.

2. The multi-antioxidant composition according to claim 1, wherein: In parts by weight, the multiple antioxidant composition comprises: 0.1-0.5 parts of recombinant collagen and 1.5-10 parts of antioxidant embedding solution.

3. The multi-antioxidant composition according to claim 1 or 2, wherein: The mass ratio of hydroxytyrosol, ergothioneine and astaxanthin in the antioxidant embedding solution is (0.1-5):(1-5):(0.05-3); Optionally, the mass ratio of hydroxytyrosol, ergothioneine and astaxanthin in the antioxidant embedding solution is (1-3):(3-5):(0.1-2).

4. The multi-antioxidant composition according to any one of claims 1 to 3, wherein: The antioxidant embedding solution is prepared by the following method, which comprises: (1) mixing hydroxytyrosol, ergothioneine and astaxanthin with linseed oil, heating and ultrasonically dispersing to obtain an oil phase; dissolving a soapberry bark extract in a phosphate buffer to obtain an aqueous phase; and mixing the oil phase and the aqueous phase to obtain a primary emulsion; (2) uniformly mixing the primary emulsion with a schizophyllan solution and a phosphate buffer to obtain a secondary emulsion; (3) The secondary emulsion is mixed evenly with pectin solution and phosphate buffer to obtain the antioxidant embedding solution.

5. The multi-antioxidant composition according to claim 4, wherein: The heating temperature is 45-55°C and the heating time is 1.5-2.5h; Optionally, the ultrasound duration is 2-5 min; Optionally, the mass ratio of the oil phase to the water phase in the primary emulsion is 1:(8-12); Optionally, the mass percentage of the Quillaja bark extract in the aqueous phase is 0.8-1.2%; Optionally, the mass percentage of the total amount of hydroxytyrosol, ergothioneine and astaxanthin in the oil phase is 10-20%.

6. The multi-antioxidant composition according to claim 4 or 5, wherein: The mass percentage of Schizophyllan in the Schizophyllan solution is 0.8-1.2%; Optionally, the solvent of the Schizophyllan solution is an acetic acid aqueous solution, and the mass percentage of acetic acid in the acetic acid aqueous solution is 0.8-1.2%.

7. The multi-antioxidant composition according to any one of claims 4 to 6, wherein: The mass percentage of pectin in the pectin solution is 3-5%; Optionally, the solvent of the pectin solution is phosphate buffer.

8. A method for preparing a multi-antioxidant composition containing recombinant collagen according to any one of claims 1 to 7, comprising: The recombinant collagen and the antioxidant embedding solution are mixed evenly to obtain the product.

9. Use of the multi-antioxidant composition containing recombinant collagen as claimed in any one of claims 1 to 7 in cosmetics.

10. A multi-antioxidant moisturizing lotion, the raw materials of which include: The multi-antioxidant composition containing recombinant collagen according to any one of claims 1 to 7, an emollient, a humectant, an emulsifier and a thickener.

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