An antioxidant composition and its use, products

Abstract

The application provides an antioxidant composition and application and product thereof, and belongs to the technical field of cosmetics. The antioxidant composition is composed of propolis and mussel mucin, and the mass ratio of the propolis and the mussel mucin is 0.25-1:1. In the antioxidant composition, the propolis and the mussel mucin play a significant synergistic effect in the aspects of antioxidation and moisturization in a specific ratio range, and the product containing the composition has the characteristics of good uniformity, high temperature resistance, cold resistance stability and the like, and has a wide application in skin care products.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology and provides an antioxidant composition and its application and products. Background Technology

[0002] Propolis is a gelatinous natural product whose main active components are flavonoids, phenolic acids, and terpenes. Studies have shown that propolis possesses various biological activities, including antioxidant, immune-boosting, antiviral, antibacterial, blood sugar-lowering, blood lipid-lowering, liver-protective, anticancer, and anti-inflammatory effects, and is widely used in food, pharmaceuticals, and cosmetics. Among these, polyphenols, flavonoids, and terpenes exhibit antioxidant activity, the mechanisms of which are related to scavenging free radicals, chelating metal ions, and inhibiting oxidase activity, thus demonstrating strong antioxidant activity.

[0003] Chinese invention patent application CN1600322A (publication date March 30, 2005) discloses a propolis composition comprising 1-80 parts of propolis extract and 20-95 parts of polyethylene glycol. Polyethylene glycol serves as a carrier, supporting the water-soluble liquid or solid dispersion. The product has a high propolis content and exhibits significant pharmacological effects such as antioxidant, lipid-lowering, beautifying, bactericidal, and antiviral properties. It can be formulated into dosage forms such as water-soluble drops, soft capsules, tablets, hard capsules, creams, suppositories, pellets, and injections.

[0004] However, the antioxidant effect of propolis extract when used alone needs further improvement, and its high viscosity generally leads to poor uniformity during use. Existing technology (Bi Jingxiu, He Lizhong, Gao Min, et al. Study on Oxidative Crosslinking Characteristics of Mussel Adhesive Protein [J]. World's Latest Medical Information Digest. 2018, Vol. 18, No. 55) discloses mussel adhesive protein (MAP) secreted byssal threads from marine mussels. This protein possesses high strength, high toughness, high adhesion, and water resistance, and is considered an excellent broad-spectrum bioadhesive for applications in medicine and bioengineering. However, its antioxidant capacity is significantly affected by the degree of crosslinking, and current research is relatively limited. However, it is expected that its antioxidant properties can be enhanced through compounding with other ingredients, showing potential applications in antioxidant skincare products.

[0005] Furthermore, most products containing propolis extract currently exhibit poor heat and cold resistance, placing high demands on their storage environment. Therefore, it is necessary to further investigate the composition and preparation process of products containing propolis extract to comprehensively improve their antioxidant activity, uniformity, and stability, thereby fully leveraging their role in antioxidant products. Summary of the Invention

[0006] The purpose of this invention is to provide:

[0007] An antioxidant composition and related technology are provided to address the technical problems of poor uniformity and low antioxidant activity in existing propolis-containing antioxidant products, or a combination thereof.

[0008] Terminology Explanation: Unless otherwise defined, all technical terms in this document have the same meaning as commonly understood by one of ordinary skill in the art to which the subject matter of the claims pertains. Unless otherwise stated, all patents, patent inventions, and disclosures cited in this document are incorporated herein by reference in their entirety. If multiple definitions exist for terms in this document, the definitions in this chapter shall prevail.

[0009] It should be understood that the above brief description and the following detailed description are exemplary and for illustrative purposes only, and do not limit the subject matter of the invention in any way. In this invention, the singular is used in conjunction with the plural unless otherwise specifically stated. It should also be noted that, unless otherwise stated, the use of “or” or “or” means “and / or”. Furthermore, the use of the term “comprising” and other forms such as “including,” “containing,” and “contains” are not limiting.

[0010] The definition of the standard chemical term can be found in the reference "Pharmacopoeia of the People's Republic of China (2020 Edition): China Medical Science and Technology Press: May 2020: First Edition".

[0011] Unless otherwise stated, conventional methods within the scope of the art shall be used.

[0012] Unless specifically defined herein, the use of all commercially available products herein employs standard techniques. For example, it may be carried out using the manufacturer's instructions for use with the kit, or in accordance with methods known in the art or the description of this invention. The techniques and methods described herein can generally be implemented according to conventional methods well known in the art, based on the descriptions in the various summary and more specific documents cited and discussed in this specification.

[0013] The terms “optional / arbitrary” or “optionally / arbitrarily” mean that the event or situation described below may or may not occur, including both the occurrence and non-occurrence of the event or situation.

[0014] The term "propolis" (Latin name Colla) used in this article refers to a resinous substance collected by worker bees from the apical buds and exudates of plants, mixed with beeswax and secretions from the bees' mandibular glands, forming an aromatic, sticky solid. It is now included in the *Pharmacopoeia of the People's Republic of China*. It possesses antioxidant, anti-inflammatory, immunomodulatory, oral health, and wound healing properties. Its taste is bitter, pungent, and cold. It enters the spleen and stomach meridians, and is primarily used to replenish deficiencies, resolve turbidity and lipids, and quench thirst; externally, it detoxifies, reduces swelling, and promotes tissue regeneration. It is used for physical weakness and premature aging, hyperlipidemia, and diabetes; externally, it is used to treat chapped skin and burns.

[0015] The term "Mussel Adhesive Protein (MAP)" used in this article refers to the natural adhesion system secreted by the byssal glands of marine mussels. It is a class of high-molecular-weight proteins rich in DOPA (3,4-dihydroxyphenylalanine, DOPA) groups, possessing a unique ability to achieve strong, reversible adhesion in moist, saline environments. Mussel adhesive proteins are rich in amino acid residues such as glycine, serine, aspartic acid, and lysine, providing a flexible backbone, hydrophilic interface, and charged side chains, which facilitate stable interactions with the aquatic environment and various interfaces. Furthermore, the most distinctive structural feature is the high content of DOPA residues, which transform into orthoquinone structures under oxidative conditions. Orthoquinones undergo covalent addition or cross-linking reactions with amino groups, thiol groups, and other nucleophilic groups, thereby forming stable covalent networks between protein molecules and between the protein and the substrate. The dynamic balance between oxidized and unoxidized DOPA gives mussel adhesive proteins both reversible adsorption and persistent solidification characteristics in moist environments.

[0016] In a first aspect, the present invention provides an antioxidant composition comprising propolis and mussel adhesive protein, wherein the mass ratio of propolis to mussel adhesive protein is 0.25-1:1.

[0017] The mass ratio of propolis to mussel adhesive protein is 0.25-1:1, including but not limited to: 0.25:1, 0.3:1, 0.35:1, 0.4:1, 0.45:1, 0.5:1, 0.55:1, 0.6:1, 0.65:1, 0.7:1, 0.75:1, 0.8:1, 0.85:1, 0.9:1, 0.95:1, or 1:1. In this invention, by further limiting the ratio of propolis to mussel adhesive protein within this range, the product containing this composition can be guaranteed to have high antioxidant properties while further improving its moisturizing performance, and simultaneously endowing the product with higher uniformity, stability, and temperature and cold resistance.

[0018] The preferred mass ratio of propolis to mussel adhesive protein is 0.5-0.75:1, including but not limited to 0.5:1, 0.55:1, 0.6:1, 0.65:1, 0.7:1, or 0.75:1. In this invention, by further limiting the proportion of propolis and mussel adhesive protein to this ratio, it is possible to ensure that the product containing this composition simultaneously possesses high antioxidant and moisturizing properties, while also exhibiting optimal homogeneity, stability, and temperature and cold resistance.

[0019] Secondly, the present invention provides the application of any of the antioxidant compositions described above in skin care products.

[0020] The skincare products mentioned include, but are not limited to, any one of skincare water, serum, lotion, and face mask.

[0021] Thirdly, the present invention provides a topical facial mask comprising any of the antioxidant compositions described above, excipients, and water; the excipients comprising one or more of glycerin, sodium hyaluronate, sodium carboxymethyl cellulose, and ethanol.

[0022] The composition, by weight, includes: 0.095-0.55 parts of antioxidant composition, 2-3 parts of glycerin, 0.05-0.1 parts of sodium hyaluronate, 1-5 parts of sodium carboxymethyl cellulose, 1-5 parts of ethanol, and 90-110 parts of deionized water.

[0023] Fourthly, the present invention provides a method for preparing any of the above-described topical facial masks, comprising the following steps: (1) After crushing propolis into powder, add anhydrous ethanol, cool and extract to obtain propolis extract; (2) Dissolve the propolis extract in anhydrous ethanol to obtain solution 1; (3) Add glycerol to water and add solution 1 while stirring to obtain solution 2; (4) Add mussel adhesive protein, sodium hyaluronate and sodium carboxymethyl cellulose to solution 2 in sequence, and stir until sodium carboxymethyl cellulose is fully swollen to obtain the solution.

[0024] Furthermore, in step (1), the temperature for cooling extraction is -10~0℃, and the time is 8-12h.

[0025] Furthermore, in step (4), the stirring speed is 1000-1500 rpm and the time is 2-4 h.

[0026] The technical feature "the mass ratio of propolis to mussel adhesive protein is 0.25-1:1" is derived from the foregoing explanation and / or the actual amounts of propolis and mussel adhesive protein used in masks 1, 4, 6, and 7 in the embodiments. Therefore, those skilled in the art can reasonably infer that the technical feature of the mass ratio of propolis to mussel adhesive protein being 0.25-1:1, its subordinate concepts, substantially equivalent technical means, and technical means that can replace this technical feature based on existing technology and conventional technical means and common knowledge should all fall within the scope of protection of this invention.

[0027] The technical feature "the mass ratio of propolis to mussel adhesive protein is 0.5:1" corresponds to the actual amount of propolis and mussel adhesive protein used in mask 1 in the embodiment. Therefore, those skilled in the art can reasonably infer that the technical feature of the mass ratio of propolis to mussel adhesive protein being 0.5:1, its subordinate concepts, its substantially equivalent technical means, and technical means that can replace this technical feature based on existing technology and conventional technical means and common knowledge should all fall within the protection scope of this invention. For example, replacing propolis or mussel adhesive protein with other commercially available propolis and mussel adhesive proteins while keeping other technical features unchanged still falls within the protection scope of this invention.

[0028] Compared with the prior art, the present invention has the following beneficial effects: (1) This invention verifies that propolis and mussel adhesive protein have a significant synergistic effect in antioxidation within a specific ratio range; at the same time, by further optimizing the ratio of the two, the moisturizing effect of the composition is further improved, and it has a wide range of applications in skin care products. (2) By further limiting the ratio of propolis and mussel adhesive protein, this invention endows the skin care product with better uniformity, stability and temperature and cold resistance, avoids the precipitation of effective ingredients, ensures the antioxidant and moisturizing effects of the product, and extends the shelf life of the product, which is of great value to actual production. Attached Figure Description

[0029] Figure 1 The bar chart shows the scavenging rate of DPPH free radicals by the topical mask prepared in this invention. Figure 2 The bar chart shows the scavenging rate of ABTS free radicals by the topical mask prepared in this invention. Figure 3 A bar chart showing the moisturizing rate of the facial mask prepared according to the present invention. Figure 4 A bar chart showing the hemolysis rate of the topical facial mask prepared according to the present invention; Figures 1-4 In the bar chart, if the letters above the bars are the same, it indicates that there is no significant difference; if the letters are different, it indicates that there is a significant difference. Detailed Implementation

[0030] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes should also fall within the scope of protection claimed by the present invention.

[0031] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all instruments, devices, equipment, reagents, products, etc., used in the embodiments of the present invention are obtained through conventional commercial means.

[0032] The propolis was purchased from Ningbo Jieshun Biotechnology Co., Ltd. The mussel adhesive protein (product specification 98%) was purchased from Xinyang Mufan Biotechnology Co., Ltd.

[0033] Example 1: This embodiment provides a series of cream-type face masks, the composition of which is shown in Table 1.

[0034] The preparation methods for mask 1 and masks 4-6 are as follows: Step 1: Weigh out the following ingredients according to the weight proportions in Table 1: propolis, mussel adhesive protein, glycerin, sodium hyaluronate, sodium carboxymethyl cellulose, anhydrous ethanol, and deionized water. Step 2: Crush the propolis with a crusher to obtain propolis powder, and set aside; Step 3: Add 10 times the mass of anhydrous ethanol to the propolis powder, stir well, and then sonicate for 30 minutes to accelerate dissolution, resulting in a dark brown propolis solution. Then, place the solution in a refrigerator at -4°C for 12 hours for cold extraction. Step 4: Filter the extracted propolis solution three times to remove impurities, evaporate the filtrate by rotary evaporation to remove most of the solvent, and then freeze-dry to obtain propolis extract; Step 5: Dissolve the propolis extract in the prescribed amount of anhydrous ethanol and set aside. Step 6: Add deionized water and glycerin to a beaker and stir well. While stirring rapidly (1000 rpm), add propolis extract dropwise to the beaker until the solution turns pale yellow. Step 7: Add mussel adhesive protein, sodium hyaluronate and sodium carboxymethyl cellulose to the above solution in sequence, and stir for 3 hours until the sodium carboxymethyl cellulose is fully swollen to obtain a spreadable face mask.

[0035] Table 1. Composition of raw materials for facial mask preparation (unit: parts by weight)

[0036] The preparation method of mask 2 is as follows: Step 1: Weigh out the following ingredients according to the proportions in Table 1: propolis, mussel adhesive protein, glycerin, sodium hyaluronate, sodium carboxymethyl cellulose, ethanol, and deionized water. Step 2: Crush the propolis with a crusher to obtain propolis powder, and set aside; Step 3: Add 10 times the mass of anhydrous ethanol to the propolis powder, stir well, and then sonicate for 30 minutes to accelerate dissolution, resulting in a dark brown propolis solution. Then, place the solution in a refrigerator at -4°C for 12 hours for cold extraction. Step 4: Filter the extracted propolis solution three times to remove impurities, evaporate the filtrate by rotary evaporation to remove most of the solvent, and then freeze-dry to obtain propolis extract; Step 5: Dissolve the propolis extract in the prescribed amount of anhydrous ethanol and set aside. Step 6: Add deionized water and glycerin to a beaker and stir well. While stirring rapidly (1000 rpm), add propolis extract dropwise to the beaker until the solution turns pale yellow. Step 7: Add sodium hyaluronate and sodium carboxymethyl cellulose to the above solution in sequence, and stir for 3 hours until the sodium carboxymethyl cellulose is fully swollen to obtain a topical mask.

[0037] The preparation method of mask 3 is as follows: Step 1: Weigh out the mussel adhesive protein, glycerin, sodium hyaluronate, sodium carboxymethyl cellulose, ethanol and deionized water according to the mass proportions in Table 1; Step 2: Add deionized water and glycerin to a beaker and stir well; Step 3: Add mussel adhesive protein, sodium hyaluronate and sodium carboxymethyl cellulose to the above solution in sequence, and stir for 3 hours until the sodium carboxymethyl cellulose is fully swollen to obtain a spreadable face mask.

[0038] Test Example 1: Mask Efficacy Test (1) Antioxidant properties Under light-protected conditions, 1 mL of the facial mask sample was directly added to 3 mL of DPPH solution (0.04 mg / mL) and mixed thoroughly to form the sample solution. 3 mL of anhydrous ethanol was used as the control group instead of the DPPH solution, and 1 mL of deionized water was used as the blank group instead of the facial mask sample. The three solutions were then incubated at 37°C for 30 min in a constant temperature shaking incubator. The absorbance of the mixed solution was measured at 517 nm using a microplate reader (Spectramax 190, Molecular Devices), and the clearance rate was calculated using the following formula:

[0039] In the formula, A0 represents the absorbance of the mixture of DPPH solution and deionized water; A1 represents the absorbance of the mixture of DPPH solution and mask sample; and A2 represents the absorbance of the mixture of mask sample and anhydrous ethanol.

[0040] 0.8 mL of the facial mask sample was added to 3.2 mL of ABTS solution (7 mM) to create the sample group solution. 0.8 mL of deionized water was used instead of the facial mask sample as the blank group. Both mixed solutions were incubated in the dark for 6 minutes. The absorbance of the mixed solutions was then measured at 734 nm using a microplate reader (Spectramax 190, Molecular Devices), and the clearance rate was calculated using the following formula:

[0041] In the formula, A0 represents the absorbance of the ABTS solution mixed with deionized water; A1 represents the absorbance of the ABTS solution mixed with the mask sample.

[0042] The above experiments were all repeated 3 times, and the results are shown in the figure. Figure 1 and Figure 2 .

[0043] (2) Moisturizing test A 2cm x 4cm piece of medical tape was flatly applied to a glass slide to simulate skin. A 0.5g sample of the facial mask was then evenly spread onto the tape, and the initial mass of the entire device was measured. The slide was then placed in an environment of 25℃ and 75%RH for 30 minutes, and the mass of the device was measured again. The moisturizing rate was calculated using the following formula:

[0044] In the formula, m0 is the total weight of the device without the addition of sample, in g; m1 is the total weight of the mask sample and device at 0 min, in g; and m2 is the total weight of the mask and device at 30 min, in g.

[0045] The experiment was repeated 3 times, and the results are shown below. Figure 3 .

[0046] (3) Blood compatibility test Mix rat red blood cells with PBS solution, centrifuge (3000 rpm, 10 min), remove the supernatant, and repeat centrifugation 3-5 times until the supernatant is clear and transparent to obtain the red blood cell solution for later use.

[0047] Take 1 mL of the mask sample and add it to 19 mL of deionized water. Mix at 37°C for 6 hours to prepare an extract.

[0048] A mask sample group was prepared by mixing 10 μL of red blood cell solution with 60 μL of mask extract and 930 μL of PBS solution. A positive control group was prepared by mixing 10 μL of red blood cell solution with 990 μL of deionized water, and a negative control group was prepared by mixing 10 μL of red blood cells with 990 μL of PBS solution. After incubating each sample group at 37°C for 3 hours, they were centrifuged (3000 rpm, 10 min). The absorbance of the supernatant at 545 nm was measured using a ELISA reader (Spectramax 190, Molecular Devices), and the hemolysis rate was calculated using the following formula:

[0049] In the formula, A + A represents the absorbance of the positive group sample. - A represents the absorbance of the negative control group; A represents the absorbance of the mask sample group.

[0050] The experiment was repeated 3 times, and the results are shown below. Figure 4 .

[0051] (4) Experimental results: according to Figure 1 , 2 The results showed that masks 1, 2, 4, 6, and 7 achieved a DPPH free radical scavenging rate of over 65% and an ABTS free radical scavenging rate of over 70%. Overall, masks 1, 4, 6, and 7 exhibited excellent antioxidant properties. The ratio of propolis to mussel adhesive protein in these masks ranged from 0.25 to 1:1, indicating that compared to using propolis and mussel adhesive protein alone, the combination of propolis and mussel adhesive protein within this ratio range had a significant synergistic effect in terms of antioxidant properties. Among them, masks 1 and 4 also achieved a moisturizing rate of over 60%, resulting in better overall skincare effects. Furthermore, mask 1 achieved a DPPH scavenging rate of 74.85±0.59%, an ABTS scavenging rate of 84.40±0.57%, and a moisturizing rate of 62.22±2.12%, demonstrating the best skincare effect.

[0052] Furthermore, as a facial product, face masks can easily come into contact with sensitive areas such as mucous membranes and eyes. Excessive irritation may cause symptoms such as bleeding and allergies. The face masks of this invention exhibit good blood compatibility in blood compatibility tests, with hemolysis rates all below 5%, indicating that these masks are non-irritating and non-corrosive to the skin and eyes when used.

[0053] The data above shows that propolis and mussel adhesive protein have synergistic antioxidant and moisturizing effects within a specific ratio range. When used as the main active ingredients in cream masks, they can endow the masks with excellent antioxidant, moisturizing and safety properties.

[0054] Test Example 2: Physicochemical Properties Test of Facial Masks A mask sample with a uniform texture and no floating matter or sediment is considered to have good homogeneity.

[0055] The mask samples prepared in the comparative examples were centrifuged at high speed (10,000 rpm, 10 min) and stored at 45°C and -20°C for 24 h respectively. After being restored to room temperature, the mask samples were observed to show no layering, discoloration or solid precipitation, which was considered to indicate that the samples had good stability.

[0056] The results are shown in Table 2.

[0057] Table 2 Physicochemical properties of antioxidant composition face masks

[0058] Propolis has low solubility in aqueous solutions, and precipitation occurs when the propolis content is too high. As shown in Table 2, propolis precipitation occurs in masks 2, 3, and 7. The precipitate forms a solid after centrifugation, indicating uneven texture. Propolis extract contains a large amount of polyphenols and flavonoids; precipitation affects the effectiveness of its active substances. Masks 1 and 4-6, on the other hand, have a uniform and stable texture, good cold and heat resistance, and can withstand the effects of environmental changes. Considering the antioxidant, moisturizing, and other effects of the masks, as well as their properties, the masks of this invention preferably contain a propolis and mussel adhesive protein ratio of 0.25-0.075:1.

[0059] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. An antioxidant composition, characterized in that, It is composed of propolis and mussel adhesive protein, wherein the mass ratio of propolis to mussel adhesive protein is 0.25-1:

1.

2. The antioxidant composition according to claim 1, characterized in that, The mass ratio of propolis to mussel adhesive protein is 0.5-0.75:

1.

3. The antioxidant composition according to claim 2, characterized in that, The mass ratio of propolis to mussel adhesive protein is 0.5:

1.

4. The use of the antioxidant composition according to any one of claims 1-3 in skin care products.

5. The application according to claim 4, characterized in that, The skincare products include any one of the following: toner, serum, lotion, and mask.

6. A type of cream-applied facial mask, characterized in that, The composition includes the antioxidant composition according to any one of claims 1-3, excipients, and water; the excipients include one or more of glycerin, sodium hyaluronate, sodium carboxymethyl cellulose, and ethanol.

7. The cream-type facial mask according to claim 6, characterized in that, The composition comprises, by weight, 0.095-0.55 parts of antioxidant composition, 2-3 parts of glycerin, 0.05-0.1 parts of sodium hyaluronate, 1-5 parts of sodium carboxymethyl cellulose, 1-5 parts of ethanol, and 90-110 parts of deionized water.

8. The method for preparing a cream-type facial mask according to claim 6 or 7, characterized in that, Includes the following steps: (1) After crushing propolis into powder, add anhydrous ethanol, cool and extract to obtain propolis extract; (2) Dissolve the propolis extract in anhydrous ethanol to obtain solution 1; (3) Add glycerol to water and add solution 1 while stirring to obtain solution 2; (4) Add mussel adhesive protein, sodium hyaluronate and sodium carboxymethyl cellulose to solution 2 in sequence, and stir until sodium carboxymethyl cellulose is fully swollen to obtain the solution.

9. The preparation method according to claim 8, characterized in that, In step (1), the temperature for cooling extraction is -10~0℃, and the time is 8-12h.

10. The preparation method according to claim 8, characterized in that, In step (4), the stirring speed is 1000-1500 rpm and the time is 2-4 hours.

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