Antioxidant composition and application thereof

Abstract

The invention provides an antioxidant composition containing pyrus ussuriensis fruit fermentation liquor and application of the antioxidant composition, and relates to the technical field of cosmetics. According to the present invention, the research results show that the composition of the Pyrus ussuriensis fruit fermentation broth and the hyaluronic acid or the hyaluronic acid salt has excellent synergistic anti-oxidation effect, such that the anti-oxidation effect of each component is reduced so as to achieve the effect of production cost reducing. In addition, the composition is high in safety and has no potential adverse reaction to a human body.

Description

Technical Field

[0001] This application relates to the field of cosmetic technology, and more particularly to an antioxidant composition and its application. Background Technology

[0002] The skin is the outermost layer of the human body and the first line of defense against the external environment. However, with the passage of time and environmental influences, the skin gradually ages, and free radicals are one of the main causes of skin aging. Free radicals can be divided into two main categories: endogenous and exogenous. Endogenous free radicals are mainly produced by the body's metabolism and exercise, while exogenous free radicals originate from ultraviolet radiation, pollution, stress, etc. They damage collagen and elastin fibers, leading to deeper wrinkles, sagging skin, and decreased barrier function. At the same time, free radicals can also oxidize subcutaneous unsaturated fatty acids to form lipofuscin, causing sunspots, melasma, age spots, and other skin blemishes.

[0003] Previous research (Chinese patent CN117165459A) found that the fermentation filtrate of pear fruit at 2%, 5%, and 10% concentrations can not only repair oxidative damage caused by ultraviolet radiation, but also provide oxidative protection against ultraviolet radiation.

[0004] How to better inhibit the generation of free radicals and thus prevent or alleviate the damage of free radicals to the skin is a technical problem that urgently needs to be solved in this field. Summary of the Invention

[0005] In order to enhance the antioxidant advantages of pear fermentation filtrate, the inventors of this application, after in-depth research, discovered for the first time that combining hyaluronic acid or its salt with a molecular weight of less than 10 kDa, which has almost no antioxidant effect, with pear fermentation filtrate can achieve a synergistic effect on the antioxidant activity, thus completing this invention.

[0006] On one hand, this application provides an antioxidant composition comprising, in a physiologically acceptable medium:

[0007] (a) At least one hyaluronic acid or its salt with a molecular weight of less than 10 kDa;

[0008] (b) Fermentation liquid of autumn pear (Pyrus ussuriensis);

[0009] The mass ratio of (a) to (b) is 1:(25-400).

[0010] The molecular weight of (a) can be any value among 379 Da, 400 Da, 500 Da, 600 Da, 700 Da, 761 Da, 800 Da, 900 Da, 1 kDa, 1.901 Da, 2 kDa, 3 kDa, 4 kDa, 5 kDa, 6 kDa, 7 kDa, 8 kDa, 8.812 kDa, 9 kDa, 9.9 kDa, 9.99 kDa, and 10 kDa.

[0011] The mass ratio of (a) to (b) can be any value among 1:25, 1:26, 1:27, 1:28, 1:29, 1:30, 1:40, 1:50, 1:60, 1:70, 1:80, 1:90, 1:100, 1:150, 1:200, 1:250, 1:300, 1:350, and 1:400.

[0012] Furthermore, the molecular weight of (a) is 379-10 kDa.

[0013] Preferably, the molecular weight of (a) is 761 Da-8812 Da.

[0014] Furthermore, the hyaluronic acid or its salt includes one or more of hyaluronic acid, sodium hyaluronate, potassium hyaluronate, and calcium hyaluronic acid.

[0015] Preferably, the hyaluronic acid or its salt is sodium hyaluronate.

[0016] Furthermore, the mass ratio of (a) to (b) is 1:(30-400).

[0017] Specifically, the fermentation broth of the autumn pear (Pyrus ussuriensis) is the fermentation filtrate of the pear fruit.

[0018] Specifically, the fermentation broth of Pyrus ussuriensis fruit is a filtrate of grape juice fermented with Hansenula polymorpha yeast / Pyrus ussuriensis fruit.

[0019] Furthermore, the pear fermentation filtrate is obtained by fermenting pears with grape juice containing Hansenula polysaccharide.

[0020] Furthermore, the fermentation broth of the autumn pear (Pyrus ussuriensis) is obtained by fermenting grape juice with Hansenula polymorpha yeast.

[0021] Preferably, the grape juice Hansenula polymorpha is grape juice Hansenula polymorpha ZS02.

[0022] Preferably, the fermentation includes fermentation at a temperature of 25℃-40℃ and a fermentation speed of 100-300 rpm for 40-60 hours.

[0023] Specifically, the fermentation temperature can be any of the following values: 25℃, 26℃, 27℃, 28℃, 29℃, 30℃, 31℃, 32℃, 33℃, 34℃, 35℃, 36℃, 37℃, 38℃, 39℃, and 40℃. The fermentation speed can be 100 rpm, 110 rpm, 120 rpm, 130 rpm, 140 rpm, 150 rpm, 160 rpm, 170 rpm, 180 rpm, 190 rpm, 200 rpm, and 210 rpm. The fermentation time can be any of the following speed values: 220rpm, 230rpm, 240rpm, 250rpm, 260rpm, 270rpm, 280rpm, 290rpm, 300rpm, and 40h, 41h, 42h, 43h, 44h, 45h, 46h, 47h, 48h, 49h, 50h, 51h, 52h, 53h, 54h, 55h, 56h, 57h, 58h, 59h, 60h, and 40h.

[0024] In a preferred embodiment, the pear fermentation filtrate can be prepared by referring to the method described in Chinese Patent CN117165459A.

[0025] In a preferred embodiment, the method for preparing the pear fermentation filtrate includes:

[0026] Step 1: Crush and filter the pears to obtain pear juice;

[0027] Step 2: Sterilize the original juice of pears to obtain the fermentation medium;

[0028] Step 3: Inoculate the grape juice with Hanseniaspora uvarum ZS02 into the fermentation medium and ferment for 40-60 hours at 25℃-40℃ and 100-300rpm to obtain pear fermentation filtrate.

[0029] Preferably, the preparation method may further include the steps of filtering and removing impurities from the pear fermentation filtrate.

[0030] The grape juice contained Hanseniaspora uvarum ZS02, which was deposited on November 7, 2022, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCCNO.26056 and address at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.

[0031] In a preferred embodiment, the method for preparing the pear fermentation product filtrate includes:

[0032] 1) Crush the ripe pears, filter them with gauze to obtain the juice, and then filter them sequentially through a 2.0μm clarifying plate, a 0.8μm clarifying plate, and a 0.22μm filter membrane to obtain the original pear juice;

[0033] 2) Sterilize the original juice of pear fruit at 100℃ for 10 minutes to obtain pear fruit juice filtrate;

[0034] 3) Inoculate a single colony of Hanseniaspora uvarum ZS02 into the grape juice filtrate and culture it in a shake flask at 200 rpm and 30℃ for 48 h to obtain the grape juice fermentation filtrate.

[0035] 4) Centrifuge the pear fermentation filtrate at 6000 rpm for 10 min, filter to remove bacteria, take the supernatant and sterilize at 80℃ for 1 h, then filter with a 0.22 μm filter membrane to finally obtain the pear fermentation product filtrate.

[0036] Those skilled in the art may also choose to purchase the grape juice fermentation product filtrate of *Hansenula polysaccharide* / *Pyrus ussuriensis* prepared by Bloomage Biotechnology Co., Ltd. for experiments. The source of this product is not specifically limited in this application.

[0037] Pear, also known as peeled pear, is a local cultivar of the Pyrus suriensis Maxim., belonging to the genus Pyrus in the Rosaceae family. Pears are rich in nutrients such as free acids, pectin, polysaccharides, fats, proteins, cellulose, iron, calcium, and phosphorus, earning them the reputation of being the "king of ginkgo nuts." Due to their warm nature and sweet, slightly sour taste, they possess properties that clear heat and resolve phlegm, nourish the stomach and lungs, quench thirst and relieve coughs, soften blood vessels, and alleviate hangovers. They are a green food rich in nutrients.

[0038] This application is the first to discover that pear fermentation filtrate and hyaluronic acid or its salts can achieve synergistic effects in both repairing and protecting against oxidative damage, and can better inhibit the production of free radicals, thereby preventing or alleviating the damage of free radicals to the skin.

[0039] Furthermore, the physiologically acceptable medium includes one or more of solvents, pH adjusters, preservatives, and thickeners.

[0040] This application may also include other functional ingredients that do not have a negative impact on anti-oxidation, such as hyaluronic acid or its salts.

[0041] Further, the molecular weight of the hyaluronic acid or its salt is 10.1kDa-3000kDa, specifically, it can be 10.1kDa, 20kDa, 30kDa, 40kDa, 41kDa, 50kDa, 60kDa, 70kDa, 80kDa, 90kDa, 100kDa, 200kDa, 300kDa, 400kDa, 500kDa, 600kDa, 700kDa, 800kDa, 900kDa, 998kDa, 1000kDa, 1 Any value from 100kDa, 1200kDa, 1300kDa, 1400kDa, 1500kDa, 1600kDa, 1700kDa, 1800kDa, 1870kDa, 1900kDa, 2000kDa, 2100kDa, 2200kDa, 2240kDa, 2300kDa, 2400kDa, 2500kDa, 2600kDa, 2700kDa, 2800kDa, 2900kDa, and 3000kDa.

[0042] This application may also add physiologically acceptable excipients, which may be suitable propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, flow aids, flavoring agents, suspending agents, coating materials, fragrances, anti-adhesion agents, binding agents, penetration enhancers, buffers, plasticizers, surfactants, foaming agents, defoamers, encapsulating agents, humectants, absorbents, diluents, flocculants and anti-flocculation agents, filter aids, release inhibitors, etc.

[0043] The compositions of this application can be prepared by general methods, wherein one or more physiologically acceptable media or carriers may be added. Those skilled in the art can select the media or carrier according to actual needs.

[0044] In a preferred embodiment, the medium may be water.

[0045] In a preferred embodiment, the method for preparing the composition includes: mixing (a) and (b) in a physiologically acceptable medium, wherein the mass ratio of (a) to (b) is 1:(25-400), and the molecular weight of (a) is less than 10 kDa.

[0046] One aspect of this application relates to compositions, which are any compositions capable of achieving the effects described in this application. The compositions include, but are not limited to, the simultaneous or sequential use of the components. "Simultaneous use" includes using them together in the same formulation or separately in different formulations. "Sequential use" includes using them sequentially in different formulations, with no restriction on the order of sequential use.

[0047] On the other hand, this application also provides the use of the described composition in the preparation of products for oxidative damage repair and / or oxidative damage protection.

[0048] Preferably, the oxidation refers to cell damage caused by excessive accumulation of free radicals.

[0049] Preferably, the sources of the free radicals can be divided into two main categories: endogenous and exogenous. Endogenous free radicals are mainly produced by human metabolism and exercise, while exogenous free radicals come from ultraviolet rays, pollution, stress, etc.

[0050] On the other hand, this application also provides the use of the composition in the preparation of skin antioxidant products.

[0051] Preferably, the product is a topical skin preparation.

[0052] Furthermore, the products include skin care products that prevent and / or alleviate skin aging, skin barrier damage, and skin pigmentation caused by free radicals.

[0053] Furthermore, the skin antioxidant includes preventing and / or alleviating one or more of the following: UV-induced skin aging, skin barrier damage, and skin pigmentation.

[0054] The skin's antioxidant properties are achieved by inhibiting the generation of free radicals.

[0055] In a preferred embodiment, the free radical inhibition rate can reach more than 20%, and can reach as high as 77.89±2.010%.

[0056] On the other hand, this application also provides a skin care product comprising the composition.

[0057] Preferably, the product or composition is mild and non-irritating to the human body and meets the requirements of cosmetic safety technical specifications.

[0058] Those skilled in the art can adjust the concentration of the composition used in this application according to the actual situation, and no specific limitation is made in this application.

[0059] In a preferred embodiment, the concentration of the fermented Pyrus ussuriensis fruit broth in the product, by mass percentage, can be 0.01%-50%, for example, any value or range of 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 20%, 30%, 40%, 50%.

[0060] In a preferred embodiment, the concentration of hyaluronic acid or its salt in the product, by mass percentage, can be 0.001%-5%, for example, any value or range of 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%.

[0061] The present invention has the following beneficial effects:

[0062] This application is the first to discover that pear fermentation filtrate and hyaluronic acid or its salts can achieve synergistic effects in both repairing and protecting against oxidative damage, resulting in more significant antioxidant efficacy.

[0063] Furthermore, even if the amount of each substance added is insufficient to exert its effect individually, the combination method provided in this application can achieve the dual function of oxidative damage repair and oxidative damage protection. By utilizing the synergistic effect between the components, the effective dosage of each component for antioxidant effect is reduced, thereby achieving the effect of reducing production costs.

[0064] In addition, the composition is highly safe and has no potential adverse reactions in humans. Detailed Implementation

[0065] To more clearly illustrate the overall concept of this application, a detailed description is provided below by way of embodiments. Numerous specific details are set forth in the following description to provide a more thorough understanding of the invention. However, it will be apparent to those skilled in the art that the invention can be practiced without one or more of these details. In other instances, certain technical features well-known in the art have not been described to avoid confusion with the invention.

[0066] It should be noted that the following detailed descriptions are illustrative and intended to provide further explanation of this application. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains.

[0067] It should be noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the exemplary embodiments according to this application. As used herein, the singular form is intended to include the plural form as well, unless the context clearly indicates otherwise. Furthermore, it should be understood that when the terms "comprising" and / or "including" are used in this specification, they indicate the presence of features, steps, operations, devices, components, and / or combinations thereof.

[0068] Unless otherwise specified, in the following embodiments, reagents or instruments whose manufacturers are not indicated are all conventional products that can be purchased commercially.

[0069] The experimental materials involved included: grape juice Hansenula polymorpha / Pyrus pyrifolia fermentation product filtrate (Pyrus pyrifolia fermentation filtrate, batch number: 20220706, Bloomage Biotechnology Co., Ltd.), sodium hyaluronate (761 Daltons, batch number: J201230755; 1901 Daltons, batch number: 21102143; 8812 Daltons, batch number: 22032442; 2.24 million Daltons, batch number: 201230411, Bloomage Biotechnology Co., Ltd.), human skin keratinocytes HaCaT, trypsin (KGI Biotechnology), ROS detection kit (KGI Biotechnology), Lactobacillus / Pyrus pyrifolia fermentation product filtrate purchased from Guangzhou Tongyi Chemical Co., Ltd., and serum-free culture medium purchased from KGI Biotechnology.

[0070] The instruments and equipment mentioned below include: a clean bench (Beijing Donglian Haer Instrument Manufacturing Co., Ltd., SCB-1520), a carbon dioxide incubator (SANYO), a digital display constant temperature water bath (Jintan Zhongda Instrument Factory), a constant temperature microplate rapid shaker (Haimen Qilin Medical Instrument Factory), a UV-8 ultraviolet lamp box (Beijing Electric Light Source Research Institute), an ST-513 ultraviolet measuring instrument (Taiwan Xianchi), and a flow cytometer (BD).

[0071] Before further describing specific embodiments of the present invention, it should be understood that the scope of protection of the present invention is not limited to the specific embodiments described below; it should also be understood that the terminology used in the embodiments of the present invention is for describing specific embodiments and not for limiting the scope of protection of the present invention. Test methods in the following embodiments that do not specify specific conditions are generally performed under conventional conditions or as recommended by the respective manufacturers.

[0072] When numerical ranges are given in the embodiments, it should be understood that, unless otherwise stated in the present invention, both endpoints of each numerical range and any value between the two endpoints may be selected. Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art. In addition to the specific methods, apparatus, and materials used in the embodiments, based on the knowledge of the prior art possessed by one of ordinary skill in the art and the description of this invention, any prior art methods, apparatus, and materials similar to or equivalent to those described, apparatus, and materials in the embodiments of this invention may be used to implement the present invention.

[0073] Unless otherwise stated, the experimental methods, detection methods, and preparation methods disclosed in this invention all employ conventional techniques in the fields of microbiology, biochemistry, analytical chemistry, cell culture, and related areas.

[0074] In addition, the "water" mentioned in this invention includes any feasible water that can be used in the art, such as deionized water, distilled water, ion-exchanged water, double-distilled water, high-purity water, and purified water.

[0075] In the following examples, unless otherwise specified, % means wt%, that is, weight percentage, and parts means mass parts.

[0076] Experiment Example 1: Antioxidant Effect of Skin Cells

[0077] 1. Sample preparation

[0078] Fermentation filtrate: The filtrate of grape juice fermented with Hansenula polysaccharide / Pyrus ussuriensis or lactobacillus / pear juice fermented with pyrethrum was added to serum-free culture medium, mixed and stirred until completely dissolved, to obtain fermentation filtrates of different concentrations (0.2%, 0.3%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%), which were then filtered through a 0.22μm filter membrane for sterilization.

[0079] Sodium hyaluronate: Sodium hyaluronate with different molecular weights (1901 Daltons, 2.24 million Daltons, 761 Daltons, 8812 Daltons) was added to serum-free culture medium, mixed and stirred until completely dissolved, to obtain a 0.01% concentration of sodium hyaluronate, which was then filtered through a 0.22μm filter membrane for sterilization.

[0080] Composition: Prepare according to the contents in Table 1. Add sodium hyaluronate, grape juice Hansenula polysaccharide / Pyrus suriensis fruit fermentation product filtrate, and Lactobacillus / pear juice fermentation product filtrate to serum-free culture medium, mix and stir until completely dissolved to obtain sample solution, and filter sterilize through a 0.22μm filter membrane.

[0081] Table 1

[0082]

[0083] 2. Oxidative damage repair efficacy

[0084] 2.1 Paving

[0085] HaCaT cells in the logarithmic growth phase were harvested at a concentration of 1×10⁻⁶. 5 Cells were seeded at a density of 1 / mL in 12-well culture plates, with 1 mL of cell suspension in each well, and cultured in a CO2 incubator at 37°C and 5% CO2 for 24 hours.

[0086] 2.2 Damage Model Establishment

[0087] Both the experimental and model groups were covered with plastic wrap, and UVA was applied at 2000 μW / cm². 2 Irradiation intensity for 1 hour, UVB at 700 μW / cm 2 The positive control group was irradiated with high intensity for 3 minutes, while the negative control group was covered with aluminum foil and not irradiated.

[0088] 2.3 Dosing detection

[0089] After irradiation, discard the old culture medium. Add samples to the experimental groups, and add serum-free culture medium to the model group and negative control group, 1 mL per well. Continue culturing for 24 h, then discard all culture medium and wash twice with PBS. Add 1.5 mL of DCFH-DA (dichlorofluorescein diacetate, prepared by diluting 0.375 μL DCFH-DA to 1 mL PBS) to each well, and incubate in a cell culture incubator for 30 min, mixing every 5 min to ensure probe binding. Discard the probe, wash twice with preheated serum-free culture medium, add 1 mL of serum-free culture medium to each well, and incubate at 37°C for 10 min. After washing once with PBS, digest the cells with trypsin, wash twice with PBS, resuspend in 300 μL PBS, and detect using flow cytometry, recording the average fluorescence intensity.

[0090] The fluorescence intensity of Mean FITC-A was statistically analyzed, as fluorescence intensity represents the amount of ROS produced. The fluorescence intensity of the negative control group, model group, and experimental group was also statistically analyzed. The ratio of the experimental group to the model group represents the ROS generation rate. The ROS inhibition rate was calculated as follows, and the results are shown in Table 2.

[0091] ROS inhibition rate = (1 - (average fluorescence intensity of experimental group / average fluorescence intensity of model group)) × 100%.

[0092] 2.4 Results Analysis

[0093] Compared with the negative control group, the t-test was used. "ns" indicates no statistical difference (p>0.05); "*" indicates a statistical difference (p<0.05); "**" indicates a significant statistical difference (p<0.01); "***" indicates an extremely significant statistical difference (p<0.001).

[0094] To illustrate the additive effect of the composition, we used the King's formula: Q=E(a+b) / (Ea+Eb-Ea×Eb) (where Q<0.55 indicates significant antagonism, Q=0.55-0.85 indicates antagonism, Q=0.85-1.15 indicates additive effect, and Q>1.15 indicates enhancement).

[0095] 3. Oxidative damage protection

[0096] 3.1 Paving

[0097] HaCaT cells in the logarithmic growth phase were harvested at a concentration of 1×10⁻⁶. 5 Cells were seeded at a density of 1 / mL in 12-well culture plates, with 1 mL of cell suspension in each well, and cultured in a CO2 incubator at 37°C and 5% CO2 for 24 hours.

[0098] 3.2 Sample Addition Processing

[0099] The experimental group discarded the old culture medium and added samples, while the model group and negative control group added serum-free culture medium, 1 mL per well, and continued to culture for 24 h before irradiation.

[0100] 3.3 Establishment and Detection of Damage Models

[0101] Both the experimental and model groups were covered with plastic wrap, and UVA was applied at 2000 μW / cm². 2 Irradiation intensity for 1 hour, UVB at 700 μW / cm 2 Irradiate the cells at the specified intensity for 3 min, while the negative control group is covered with aluminum foil and not irradiated. After irradiation, discard the old culture medium, add serum-free medium, and continue culturing for 3 h. After culturing, discard all culture medium and wash twice with PBS. Add 1.5 mL of DCFH-DA to each well, place in a cell culture incubator, and incubate for 30 min, mixing every 5 min to ensure complete probe binding. Discard the probe, wash twice with preheated serum-free medium, add 1 mL of serum-free medium to each well, and incubate at 37°C for 10 min. After washing once with PBS, digest the cells with trypsin, wash twice with PBS, resuspend in 300 μL of PBS, and detect using flow cytometry, recording the average fluorescence intensity.

[0102] The fluorescence intensity of Mean FITC-A was statistically analyzed, as fluorescence intensity represents the amount of ROS produced. The fluorescence intensity of the negative control group, model group, and experimental group was also statistically analyzed. The ratio of the experimental group to the model group represents the ROS generation rate. The ROS inhibition rate was calculated as follows, and the results are shown in Table 3.

[0103] ROS inhibition rate = (1 - (average fluorescence intensity of experimental group / average fluorescence intensity of model group)) × 100%.

[0104] 3.4 Results Analysis

[0105] Compared with the negative control group, the t-test was used. "ns" indicates no statistical difference (p>0.05); "*" indicates a statistical difference (p<0.05); "**" indicates a significant statistical difference (p<0.01); "***" indicates an extremely significant statistical difference (p<0.001).

[0106] To illustrate the additive effect of the composition, we used the King's formula: Q=E(a+b) / (Ea+Eb-Ea×Eb) (where Q<0.55 indicates significant antagonism, Q=0.55-0.85 indicates antagonism, Q=0.85-1.15 indicates additive effect, and Q>1.15 indicates enhancement).

[0107] Table 2 Results of UV damage repair efficacy test

[0108]

[0109] Note: The fermentation filtrate group uses the same fermentation filtrate as described in the corresponding examples or comparative examples as the sample solution; the sodium hyaluronate group uses the same sodium hyaluronate as described in the corresponding examples or comparative examples as the sample solution; and the composition group uses the compositions described in Examples 1-9 and Comparative Examples 1-4 as the sample solution.

[0110] Table 3 Results of UV damage protection efficacy test

[0111]

[0112] Note: The fermentation filtrate group uses the same fermentation filtrate as described in the corresponding examples or comparative examples as the sample solution; the sodium hyaluronate group uses the same sodium hyaluronate as described in the corresponding examples or comparative examples as the sample solution; and the composition group uses the compositions described in Examples 1-9 and Comparative Examples 1-4 as the sample solution.

[0113] Conclusion: As shown in Table 2, the combination of grape juice fermentation product filtrate of *Hansenula polysaccharide* / *Pyrus pyrifolia* and sodium hyaluronate has the effect of inhibiting ROS after UV damage. Compared with the use of grape juice fermentation product filtrate of *Hansenula polysaccharide* / *Pyrus pyrifolia* or sodium hyaluronate alone, the combined use has a more significant inhibitory effect on ROS after UV damage, and plays a synergistic role in inhibiting ROS after UV damage, thus achieving a better oxidative damage repair effect.

[0114] As shown in Table 3, the combination of grape juice Hansenula polysaccharide / Apricot fruit fermentation product filtrate and sodium hyaluronate added before UV irradiation can effectively reduce UV-induced ROS. Compared with the use of grape juice Hansenula polysaccharide / Apricot fruit fermentation product filtrate or sodium hyaluronate alone, the combined use has a more significant inhibitory effect on ROS, and plays a synergistic role in reducing UV-induced ROS, thus achieving a better protective effect against oxidative damage.

[0115] Experimental Example 2: Patch Test

[0116] The patch test in this embodiment was conducted in accordance with the "Human Skin Patch Test" in the Cosmetic Safety Technical Specifications (2015 Edition).

[0117] Instrumentation: Patch tester (Chemotechnique Diagnostics AB, Sweden).

[0118] Subject Recruitment: We are recruiting 30 healthy subjects, regardless of gender.

[0119] Sample preparation: According to the contents in Table 4, add sodium hyaluronate and grape juice Hansenula polysaccharide / Pyrus ussuriensis fruit fermentation product filtrate to purified water, mix and stir until completely dissolved to obtain the sample solution.

[0120] Preparation of the control group: Purified water was used as the control group.

[0121] Experimental Method: Tear open the packaging of the plaque applicator and measure 0.020 g of sample into the small chamber. Apply the plaque applicator to the flexor side of the subject's forearm, and gently press it with the palm of your hand to ensure even application to the skin, for 24 hours. After removing the plaque applicator, observe and record the reaction results according to Table 5 for 48 hours. The results are shown in Table 6.

[0122] Table 4

[0123]

[0124]

[0125] Table 5. Grading Criteria for Skin Reactions in Closed Patch Tests

[0126]

[0127] Table 6

[0128] Group Level 0 Level 1 Level 2 Level 3 Level 4 Sample 1 30 0 0 0 0 Sample 2 30 0 0 0 0 Sample 3 30 0 0 0 0 Sample 4 30 0 0 0 0 Sample 5 30 0 0 0 0 Sample 6 30 0 0 0 0 Sample 7 30 0 0 0 0 Sample 8 30 0 0 0 0 Sample 9 30 0 0 0 0 control group 30 0 0 0 0

[0129] Conclusion: Samples 1-9 all showed negative reactions and were mild and non-irritating to the human body.

[0130] The above description is merely an embodiment of this application and is not intended to limit the scope of this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the scope of the claims of this application.

Claims

1. An antioxidant composition, characterized in that, The composition comprises, in a physiologically acceptable medium, the following: (a) At least one hyaluronic acid or its salt with a molecular weight of less than 10 kDa; (b) Fermentation liquid of autumn pear (Pyrus ussuriensis); The mass ratio of (a) to (b) is 1:(25-400).

2. The composition according to claim 1, characterized in that, The molecular weight of (a) is 379–10 kDa.

3. The composition according to claim 1, characterized in that, The mass ratio of (a) to (b) is 1:(30-400).

4. The composition according to claim 1, characterized in that, (b) is the fermented filtrate of pear fruit.

5. The composition according to claim 4, characterized in that, (b) is obtained by fermenting grape juice with Hansenula polysaccharide yeast to obtain pear fruit.

6. Use of the composition according to any one of claims 1-5 in the preparation of products for repairing and / or protecting against oxidative damage.

7. Use of the composition according to any one of claims 1-5 in the preparation of skin antioxidant products.

8. The application according to claim 6 or 7, characterized in that, The products include skin care products that prevent and / or alleviate skin aging, skin barrier damage, and skin pigmentation caused by free radicals.

9. A skin care product, characterized in that, The skin care product comprises the composition according to any one of claims 1-5.

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