Multi-functional polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum composition

Abstract

The present invention discloses a composition of polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum with multiple functions, belonging to the technical field of cosmetics. The preparation method of the composition of polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum with multiple functions comprises the following steps: taking hydrolyzed sclerotium gum and deionized water, mixing them, performing ultrasonic oscillation, adding polyglutamic acid hydrogel and antioxidant hyaluronic acid, heating and stirring, filtering, transferring the solid phase into a template, placing the template in an environment of -20°C and freezing for 6-8 h, taking it out, melting it at room temperature for 2-3 h, and performing freeze-thawing 3-4 times repeatedly, thus obtaining the composition of polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum with moisturizing, repairing, firming and anti-aging functions.

Description

Technical Field

[0001] The present invention belongs to the technical field of cosmetics, and particularly relates to a composition of polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum with multiple functions. Background Art

[0002] Under the influence of external environmental pollutants, microbial invasion, sunlight and other stimulating factors, the skin may experience different degrees of allergies, such as symptoms like dryness, itching, stinging and tightness, as well as problems such as aging, wrinkles, discoloration and dull skin color that seriously affect skin health. Currently, there are numerous anti-aging and repair compositions on the market, but their anti-aging and repair effects are always unsatisfactory and difficult to meet people's needs. Therefore, in order to better achieve anti-aging and repair effects, it is necessary to find an anti-aging and repair composition with excellent performance. Summary of the Invention

[0003] The purpose of the present invention is to provide a composition of polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum with multiple functions for improving skin care effects.

[0004] The purpose of the present invention can be achieved by the following technical solutions:

[0005] A composition of polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum with multiple functions, and the preparation method of the composition of polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum with multiple functions includes the following steps:

[0006] Take hydrolyzed sclerotium gum and deionized water and mix them, perform ultrasonic oscillation, add polyglutamic acid hydrogel (purchased from Macheng Jinxin Biotechnology Co., Ltd.), antioxidant hyaluronic acid, heat and stir, filter, transfer the solid phase into a template, place the template in an environment at -20°C and freeze for 6 - 8 h, take it out, melt it at room temperature for 2 - 3 h, and perform freeze-thaw cycles 3 - 4 times to obtain the composition of polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum.

[0007] As a preferred technical solution of the present invention, the mass ratio of hydrolyzed sclerotium gum, deionized water, polyglutamic acid hydrogel, and antioxidant hyaluronic acid is 20 - 30:50 - 60:70 - 80:25 - 35.

[0008] As a preferred technical solution of the present invention, the molecular weight of the hydrolyzed sclerotium gum is 1.2 - 2.2 kDa.

[0009] As a preferred technical solution of the present invention, the preparation method of the antioxidant hyaluronic acid includes the following steps:

[0010] Take hyaluronic acid (purchased from Diyuan Biochemical Products Co., Ltd., Dingtao County), mix it with deionized water, stir for 10 - 12 h, add sodium periodate, stir in the dark for 2 - 3 h, dialyze and purify, and then conduct vacuum freeze-drying to obtain antioxidant hyaluronic acid.

[0011] As a preferred technical solution of the present invention, the dosage ratio of the hyaluronic acid, deionized water, and sodium periodate is 2 - 3 g : 200 - 300 mL : 20 mL.

[0012] As a preferred technical solution of the present invention, the heating and stirring refers to stirring at 30 - 40 °C for 4 - 6 h.

[0013] As a preferred technical solution of the present invention, the vacuum freeze-drying refers to vacuum freeze-drying at -50 °C for 8 - 10 h.

[0014] As a preferred technical solution of the present invention, the time of ultrasonic oscillation is 15 - 25 min.

[0015] An application of a polyglutamic acid hydrogel, hyaluronic acid, and hydrolyzed sclerotium gum composition with multiple functions, where the application refers to using the polyglutamic acid hydrogel, hyaluronic acid, and hydrolyzed sclerotium gum composition as a nutrient or a carrier for the preparation of cosmetics.

[0016] Advantages of the present invention:

[0017] A polyglutamic acid hydrogel, hyaluronic acid, and hydrolyzed sclerotium gum composition with multiple functions disclosed by the present invention is obtained by compounding hydrolyzed sclerotium gum, polyglutamic acid hydrogel, and antioxidant hyaluronic acid. Through the synergistic effect of the three, it plays roles in moisturizing, repairing, firming, and anti-aging. Specific embodiments

[0018] To further elaborate on the technical means and effects adopted by the present invention to achieve the predetermined invention purpose, the following combines examples to detail the specific embodiments, structures, features, and their effects according to the present invention as follows.

[0019] The preparation of the hydrolyzed sclerotium gum in this application specifically includes the following steps:

[0020] Inoculate Sclerotium rolfsii in a 250 mL shake flask containing 50 mL of seed fermentation medium, and culture it at 28 °C and 220 r / min for 72 h;

[0021] Take an inoculum of 5% of the volume of the seed fermentation medium and transfer it to a 250 mL shake flask containing 25 mL of glucose fermentation medium, culture it at 30 °C and 220 r / min for 96 h, conduct pressure filtration, and collect 50 mL of sclerotium rolfsii fermentation broth;

[0022] Heat the fermentation broth of *Sclerotium rolfsii* to 90°C, hold for 20 min, and then cool to room temperature;

[0023] Enzymatic hydrolysis: Adjust the pH of the fermentation broth of *Sclerotium rolfsii* to 6.0 with 10% citric acid by mass, then add a mixed enzyme composed of 0.05% lysozyme, chitinase, and hemicellulase by mass. After stirring at 45°C for 4 h, adjust the pH to 8.0 with 1 mol / L sodium hydroxide, add 0.1% papain by mass, heat up to 55°C, react for 4 h, heat to 100°C, hold for 60 min, cool to 10°C, and centrifuge at 4000 r / min to collect the supernatant;

[0024] Alcohol precipitation: Add absolute ethanol to the supernatant, with the volume ratio of the supernatant to absolute ethanol being 1:1, mix and stir for 30 min, and then centrifuge;

[0025] Purification: Collect the precipitate, redissolve it with deionized water, add 2.5 g of powdered activated carbon, hold at 100°C for 30 min, cool to room temperature, centrifuge at 13000 r / min using a high-speed centrifuge to obtain the centrifuged supernatant, filter the centrifuged supernatant through a 0.1-μm ceramic membrane, and take the filtrate to obtain hydrolyzed *Sclerotium rolfsii* gum.

[0026] The components of the seed fermentation medium are: glucose 100 g / L, NaNO₃ 3 g / L, KH₂PO₄ 1.3 g / L, citric acid 0.7 g / L, KCl 0.5 g / L, MgSO₄·7H₂O 0.5 g / L, FeSO₄·7H₂O 0.05 g / L, vitamin B₁ 3.3 g / L, ZnSO₄·7H₂O 3.3 g / L, yeast extract 1 g / L, and the pH is 4.6 (adjusted with phosphoric acid);

[0027] The components of the glucose fermentation medium are: potato 200 g / L, glucose 20 g / L, agar 15 g / L, and distilled water 1000 mL;

[0028] The mass ratio of the lysozyme, chitinase, hemicellulase, and papain is 2:1:1:0.5;

[0029] All *Sclerotium rolfsii* involved in this application are sourced from Ningbo Mingzhou Biotechnology Co., Ltd.

[0030] Example 1

[0031] A preparation method of a multi-functional composition of polyglutamic acid hydrogel, hyaluronic acid, and hydrolyzed *Sclerotium rolfsii* gum comprises the following steps:

[0032] Mix hydrolyzed sclerotium gum and deionized water, and ultrasonically oscillate for 15 min. Then add polyglutamic acid hydrogel and antioxidant hyaluronic acid, and stir at 30 °C for 4 h. Filter, transfer the solid phase into a template, place the template in an environment at -20 °C and freeze for 6 h. Take it out and melt it at room temperature for 2 h. Repeat the freeze-thaw cycle 3 times to obtain the polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum composition. The mass ratio of the hydrolyzed sclerotium gum, deionized water, polyglutamic acid hydrogel and antioxidant hyaluronic acid is 20:50:70:25. The molecular weight of the hydrolyzed sclerotium gum is 1.2 kDa.

[0033] The preparation method of the antioxidant hyaluronic acid includes the following steps:

[0034] Mix hyaluronic acid and deionized water, and stir for 10 h. Then add sodium periodate and stir in the dark for 2 h. Dialyze and purify, and vacuum freeze-dry at -50 °C for 8 h to obtain antioxidant hyaluronic acid. The dosage ratio of the hyaluronic acid, deionized water and sodium periodate is 2 g:200 mL:20 mL.

[0035] Example 2

[0036] A preparation method of a polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum composition with multiple functions includes the following steps:

[0037] Mix hydrolyzed sclerotium gum and deionized water, and ultrasonically oscillate for 20 min. Then add polyglutamic acid hydrogel and antioxidant hyaluronic acid, and stir at 35 °C for 5 h. Filter, transfer the solid phase into a template, place the template in an environment at -20 °C and freeze for 7 h. Take it out and melt it at room temperature for 2.5 h. Repeat the freeze-thaw cycle 3 times to obtain the polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum composition. The mass ratio of the hydrolyzed sclerotium gum, deionized water, polyglutamic acid hydrogel and antioxidant hyaluronic acid is 25:55:75:30. The molecular weight of the hydrolyzed sclerotium gum is 1.7 kDa.

[0038] The preparation method of the antioxidant hyaluronic acid includes the following steps:

[0039] Mix hyaluronic acid and deionized water, and stir for 11 h. Then add sodium periodate and stir in the dark for 2.5 h. Dialyze and purify, and vacuum freeze-dry at -50 °C for 9 h to obtain antioxidant hyaluronic acid. The dosage ratio of the hyaluronic acid, deionized water and sodium periodate is 2.5 g:250 mL:20 mL.

[0040] Example 3

[0041] A preparation method of a polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum composition with multiple functions includes the following steps:

[0042] Mix hydrolyzed scleroglucan and deionized water, and ultrasonically oscillate for 25 min. Add polyglutamic acid hydrogel and antioxidant hyaluronic acid, and stir at 40 °C for 6 h. Filter, transfer the solid phase into a template, place the template in an environment at -20 °C and freeze for 8 h, take it out, and melt it at room temperature for 3 h. Repeat the freeze-thaw cycle 4 times to obtain the composition of polyglutamic acid hydrogel, hyaluronic acid, and hydrolyzed scleroglucan; the mass ratio of hydrolyzed scleroglucan, deionized water, polyglutamic acid hydrogel, and antioxidant hyaluronic acid is 30:60:80:35; the molecular weight of the hydrolyzed scleroglucan is 2.2 kDa;

[0043] The preparation method of the antioxidant hyaluronic acid includes the following steps:

[0044] Mix hyaluronic acid and deionized water, and stir for 12 h. Add sodium periodate, and stir in the dark for 3 h. Dialyze and purify, and vacuum freeze-dry at -50 °C for 10 h to obtain antioxidant hyaluronic acid; the dosage ratio of hyaluronic acid, deionized water, and sodium periodate is 3 g: 300 mL: 20 mL.

[0045] Comparative Example 1

[0046] The difference from Example 1 is that the mass ratio of hydrolyzed scleroglucan, deionized water, polyglutamic acid hydrogel, and antioxidant hyaluronic acid is 0:55:100:30.

[0047] Comparative Example 2

[0048] The difference from Example 1 is that the mass ratio of hydrolyzed scleroglucan, deionized water, polyglutamic acid hydrogel, and antioxidant hyaluronic acid is 25:55:0:105.

[0049] Comparative Example 3

[0050] The difference from Example 1 is that the mass ratio of hydrolyzed scleroglucan, deionized water, polyglutamic acid hydrogel, and antioxidant hyaluronic acid is 25:55:105:0.

[0051] Comparative Example 4

[0052] The difference from Example 1 is that hyaluronic acid is not modified.

[0053] Performance Test

[0054] Test Example 1

[0055] Human patch test

[0056] 1. Test substance: The product of Example 1.

[0057] 2. Negative control: Blank control.

[0058] 3. Subjects: A total of 30 people, including 3 males and 27 females, aged 18 - 59 years old, with an average age of 37.2 ± 10.6 years old, meeting the volunteer selection criteria for subjects.

[0059] 4. Patch test method: Select qualified patch test equipment. Using the closed patch test method, place approximately 0.020g - 0.025g of the test substance in the patch test device, and apply it to the back of the subject with a low - sensitization tape. After 24 hours, remove the test substance, and observe the skin reaction at 0.5, 24, and 48 hours after removal respectively. Record the results according to the skin reaction grading standard in the currently effective technical specification.

[0060] 5. Experimental results:

[0061]

[0062]

[0063] 6. Skin reaction grading standard:

[0064]

[0065] The poly - glutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum composition of this application has no adverse reactions on human skin; the inventors of this application repeated the above - mentioned test with the samples prepared in other examples and comparative examples, and the test results were the same, all having no adverse reactions on human skin.

[0066] Test Example 2

[0067] Randomly select 35 skin - healthy volunteers (the male - to - female ratio is 15:20), randomly divide them into 7 groups, with 5 people in each group. Each group corresponds to a poly - glutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum composition in an example or a comparative example;

[0068] After each volunteer cleans the facial skin with facial cleanser, dries it, then cleans the facial skin with clean water and dries it again, 12 test groups respectively take 1g of the poly - glutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum compositions prepared in Examples 1 - 3 and Comparative Examples 1 - 4, dissolve them in 100 mL of deionized water to make a paste, evenly apply it on the face, and gently pat the facial skin for 30 seconds for testing, and take the average value;

[0069] After the facial moisture dries naturally, use a skin detector to measure the moisture content A of the facial skin at room temperature of 25°C and humidity of 60%; sit quietly indoors for 1 hour, and measure the moisture content B of the facial skin again under the same conditions, and calculate the moisture retention rate. The higher the moisture retention rate, the better the water - locking performance;

[0070] After comparison, perform statistical analysis. The comparison results of each group show that P < 0.05, and the difference is statistically significant;

[0071] Water retention rate = B ÷ A × 100%

[0072] The test results are shown in Table 1 below;

[0073] Table 1

[0074] Moisture retention rate (%) Example 1 85.7 Example 2 86.8 Example 3 85.4 Comparative Example 1 69.2 Comparative Example 2 70.8 Comparative Example 3 71.3 Comparative Example 4 80.8

[0075] Test results: The polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum compositions prepared in Examples 1 - 3 have good water locking performance; the water locking performance of the polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum compositions prepared in Comparative Examples 1 - 3 has significantly decreased.

[0076] Test Example 3

[0077] Apply the polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum compositions prepared in the above Examples 1 - 3 and Comparative Examples 1 - 4 to the acne - inflamed areas of patients.

[0078] Test subjects: Randomly select 140 volunteers with acne inflammation of different degrees, aged 18 - 30 years old. Among them, there are 48 people with neutral skin type, 51 people with oily skin type, and 41 people with dry skin type.

[0079] Test method: Take 2 g of the polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum compositions prepared in the above Examples 1 - 3 and Comparative Examples 1 - 4 and apply them evenly on the acne - inflamed areas and within a radius of 3 cm. Apply once in the morning and once in the evening. After continuous use for two weeks, evaluate the use effect of the polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum compositions. The test results are shown in Table 2. Divide 20 people into 1 group, a total of 7 groups. Each group corresponds to a polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed sclerotium gum composition in an example or a comparative example.

[0080] Evaluation criteria for use effect:

[0081] Grade Ⅰ: Obvious repair effect, significant soothing effect, no irritation, soft skin feeling during use, moisturizing, moist but not greasy, the skin is moist, smooth and bright after use;

[0082] Grade Ⅱ: Good repair effect, good soothing effect, no irritation, general skin feeling during use, moisturizing, the skin moisture is average after use, and the skin gloss change is not obvious;

[0083] Grade Ⅲ: General repair effect, general soothing effect, no irritation, general skin feeling during use, general moisturizing effect, no obvious change in the skin and gloss after use;

[0084] Grade Ⅳ: Poor repair effect, poor soothing effect, slightly irritating, general skin feeling during use, general moisturizing effect, no change in the skin and gloss after use.

[0085] Table 2: Test Results of the Soothing and Repairing Effects of Polyglutamic Acid Hydrogel, Hyaluronic Acid and Sclerotium rolfsii Gum Hydrolysate Compositions in Examples 1-3 and Comparative Examples 1-4

[0086] Grade I Grade II Grade III Grade IV Example 1 19 1 0 0 Example 2 20 0 0 0 Example 3 20 0 0 0 Comparative Example 1 0 0 1 19 Comparative Example 2 2 17 1 0 Comparative Example 3 1 18 0 1 Comparative Example 4 3 15 1 1

[0087] As shown in Table 2, the polyglutamic acid hydrogel, hyaluronic acid and sclerotium rolfsii gum hydrolysate compositions of Examples 1-3 are considered to have good soothing and repairing effects, and have a good skin feel during use; while the soothing and repairing effects of the sclerotium rolfsii gum hydrolysate in Comparative Examples 1-4 are worse than those of Examples 1-3.

[0088] Test Example 4

[0089] (1) Test Subjects: 40 volunteers aged 30-55 with wrinkles and healthy and intact skin were randomly divided into 4 groups, with 10 volunteers in each group.

[0090] (2) Products Used: The polyglutamic acid hydrogel, hyaluronic acid and sclerotium rolfsii gum hydrolysate compositions prepared from the compositions in Examples 1-3 and Comparative Example 4 were used once in the morning and once in the evening every day for one cycle, and one cycle was 30 days.

[0091] (3) Usage Method: After cleansing the face in the morning and evening every day, take an appropriate amount of essence and apply it evenly on the face, and gently massage until absorbed. The volunteers were randomly divided into 4 groups, and 10 volunteers in each group were respectively used with the polyglutamic acid hydrogel, hyaluronic acid and sclerotium rolfsii gum hydrolysate compositions prepared in Examples 1-3 and Comparative Example 4.

[0092] (4) Test Items: Anti-aging test. Specifically, before and after the cyclic use, a VISIA skin detector was used to detect and analyze the use effects. The detected contents included skin texture, pores and wrinkles. The test results were expressed as percentages, representing the comparison ranking of the skin characteristics of the tested person compared with others of the same race, age and muscle type.

[0093] The average value of each group's index data was taken as the final result, and the anti-aging test was carried out on the compositions prepared in Examples 1-3 and Comparative Example 4 of the present invention. The results are shown in Table 3.

[0094] Table 3

[0095]

[0096] As can be seen from Table 3, the polyglutamic acid hydrogel, hyaluronic acid and sclerotium rolfsii gum hydrolysate compositions prepared in Examples 1-3 of the present application have good anti-aging effects.

[0097] The above are only the preferred embodiments of the present invention and do not impose any formal restrictions on the present invention. Although the present invention has been disclosed above in the preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some changes or modifications to equivalent embodiments by using the above-disclosed technical content without departing from the technical solution of the present invention. However, as long as it does not depart from the technical solution content of the present invention, any brief modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention still fall within the scope of the technical solution of the present invention.

Claims

1. A polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed scleroglucan composition with multiple functions, characterized in that, The preparation method of the composition of polyglutamic acid hydrogel with multiple functions, hyaluronic acid and sclerotium rolfsii hydrolyzate comprises the following steps: Take sclerotium rolfsii hydrolyzate and deionized water, mix them, carry out ultrasonic oscillation, add polyglutamic acid hydrogel and antioxidant hyaluronic acid, heat and stir, filter, transfer the solid phase into a template, place the template in an environment at -20°C and freeze for 6 - 8 h, take it out, melt it at room temperature for 2 - 3 h, and carry out freeze-thaw cycles for 3 - 4 times to obtain the composition of polyglutamic acid hydrogel, hyaluronic acid and sclerotium rolfsii hydrolyzate; The mass ratio of the sclerotium rolfsii hydrolyzate, deionized water, polyglutamic acid hydrogel and antioxidant hyaluronic acid is 20 - 30:50 - 60:70 - 80:25 - 35; The preparation method of the antioxidant hyaluronic acid comprises the following steps: Take hyaluronic acid and deionized water, mix them, stir for 10 - 12 h, add sodium periodate, stir in the dark for 2 - 3 h, carry out dialysis purification, and vacuum freeze-dry to obtain antioxidant hyaluronic acid; The dosage ratio of the hyaluronic acid, deionized water and sodium periodate is 2 - 3 g:200 - 300 mL:20 mL.

2. A composition of polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed scleroglucan with multiple functions according to claim 1, characterized in that, The molecular weight of the sclerotium rolfsii hydrolyzate is 1.2 - 2.2 kDa.

3. A composition of a polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed scleroglucan with multiple functions according to claim 1, characterized in that, The heating and stirring means stirring at 30 - 40°C for 4 - 6 h.

4. A composition of a polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed scleroglucan with multiple functions according to claim 1, characterized in that, The vacuum freeze-drying means vacuum freeze-drying at -50°C for 8 - 10 h.

5. A polyglutamic acid hydrogel, hyaluronic acid and hydrolyzed scleroglucan composition with multiple functions according to claim 1, characterized in that, The time of the ultrasonic oscillation is 15 - 25 min.

6. Use of a composition of a polyglutamic acid hydrogel, hyaluronic acid, and hydrolyzed scleroglucan having multiple functions according to any one of claims 1-5, characterized in that, The application refers to using the composition of polyglutamic acid hydrogel, hyaluronic acid and sclerotium rolfsii hydrolyzate as a nutrient or a carrier for the preparation of cosmetics.

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