A copolymer of tremella polysaccharide grafted with metal ions and fullerol and its preparation method and application

Abstract

The present invention relates to the technical field of cosmetic formula, and specifically discloses a tremella polysaccharide grafted metal ion and fullerol copolymer and its preparation method and application. A preparation method of a tremella polysaccharide grafted metal ion and fullerol copolymer, step one: weighing raw materials, including metal salt, tremella polysaccharide and fullerol; step two: dissolving the metal salt in deionized water to prepare a metal salt solution; step three: adding the weighed tremella polysaccharide and fullerol to the above-mentioned metal salt solution to obtain a suspension; step four: heating the suspension, reacting to the reaction end point under ultrasonic conditions, rotary evaporation to 1 / 3 of the liquid volume, cooling and drying and post-processing to obtain a tremella polysaccharide grafted metal ion and fullerol copolymer. Through the metal ion-tremella polysaccharide-fullerol raw material combination, due to the synergistic effect of the raw materials, the grafted copolymer not only exerts the natural moisturizing, anti-inflammatory and repairing properties of tremella polysaccharide, but also enhances the anti-inflammatory effect and antioxidant capacity.

Description

Technical Field

[0001] The invention relates to the technical field of cosmetic formulas, and in particular to a tremella polysaccharide grafted metal ion and fulleranol copolymer, and a preparation method and application thereof. Background Art

[0002] Tremella polysaccharide, as a natural high molecular weight polysaccharide extracted from Tremella, has attracted much attention in the cosmetics field due to its excellent moisturizing, anti-inflammatory and repairing properties. It can effectively provide moisture to the skin and promote the recovery of damaged skin. 2+ The addition of Zn helps to enhance the skin's barrier function, while 2+ As a component of many important enzymes, it plays a key role in the repair and regeneration of the skin. Tremella polysaccharide is a natural active ingredient with antioxidant, anti-aging, anti-tumor, blood pressure lowering, blood lipid lowering, immune enhancement and other functions, and is relatively safe. Tremella polysaccharide can be widely used in many fields such as medical treatment, health care, food processing, skin care and beauty, animal husbandry and breeding. Moreover, the wide source of raw materials can bring greater application space for Tremella polysaccharide, which is of great significance for further research on the physicochemical properties and biological activity of Tremella polysaccharide. With the development of polysaccharide biological activity, more and more research is devoted to the effect of polysaccharide on skin moisturizing, anti-aging or whitening. However, most of the research on Tremella polysaccharide stays in the optimization of polysaccharide extraction process, structural characterization, antioxidant, anti-cancer, blood lipid lowering, blood sugar lowering and enhancing the body's immune activity. There are few or in-depth studies on the skin care activity of Tremella polysaccharide.

[0003] CN113893182A discloses a water-soluble fullerene complex and its preparation method and application, which includes raw materials such as tremella polysaccharide, polyglutamic acid, ectoine and polyglutamic acid. Due to the hydrophobic and oleophobic properties of fullerene, the grafting rate of fullerene and tremella polysaccharide in the prepared fullerene-tremella polysaccharide mixture is not high, and the synergistic effect of fullerene on the final product is limited.

[0004] CN114272187A discloses a hydrating and moisturizing dry facial mask containing fullerene and a preparation method thereof, which comprises raw materials such as palmitoyl tripeptide, okra polysaccharide, hydrolyzed sclerotium gum, fullerene, tremella polysaccharide, etc., but fails to fully utilize the efficacy of fullerene and tremella polysaccharide.

[0005] It can be seen that although the prior art uses Tremella polysaccharide as a raw material to prepare products for use in cosmetics, how to make Tremella polysaccharide fully exert its moisturizing and repairing effects in cosmetics; as fullerene is a nanomaterial, the use of nanomaterials in cosmetics is considered unsafe, and how to solve the safety problem of the application of nanomaterial fullerene in cosmetics needs to be solved urgently. Summary of the invention

[0006] In view of the above problems, an object of the present invention is to provide a method for preparing a copolymer of tremella polysaccharide grafted with metal ions and fullerol. Through a unique raw material combination and graft copolymerization technology, metal ions such as calcium ions and zinc ions are compounded with tremella polysaccharide and fullerol to form a copolymer of tremella polysaccharide grafted with metal ions and fullerol. The preparation method is simple and ensures that fullerol is uniformly grafted on the tremella polysaccharide compound gel. The amount of fullerol added can be strictly controlled to ensure the stability, safety and efficacy of the material.

[0007] The second object of the present invention is to provide a copolymer of tremella polysaccharide grafted with metal ions and fullerol. It has good stability and safety. Not only can the respective effects be fully exerted, but also the skin care effect is significantly enhanced through synergistic effects. Not only the natural moisturizing, anti-inflammatory and repairing properties of tremella polysaccharide are retained, but also its anti-inflammatory and antioxidant abilities are significantly enhanced.

[0008] The third object of the present invention is to provide a copolymer of Tremella fuciformis polysaccharide grafted with metal ions and fullerol for use in cosmetics. Compared with single-ingredient products, it more comprehensively meets the multiple needs of modern people for cosmetics, combines anti-oxidation, anti-inflammatory, moisturizing and soothing, and realizes comprehensive skin care.

[0009] The first technical solution adopted by the present invention is: a method for preparing a copolymer of Tremella polysaccharide grafted with metal ions and fullerol, comprising:

[0010] Step 1: weighing raw materials, including metal salt, Tremella polysaccharide and fullerenol;

[0011] Step 2: dissolving the metal salt in deionized water to prepare a metal salt solution;

[0012] Step 3: adding the Tremella polysaccharide and fullerenol to the metal salt solution to obtain a suspension;

[0013] Step 4: heating the suspension, reacting to the reaction end point under ultrasonic conditions, rotary evaporating to 1 / 3 of the liquid volume, cooling, and drying to obtain the Tremella polysaccharide grafted metal ion and fullerene alcohol copolymer.

[0014] Preferably, the metal salt includes one or more of zinc gluconate, calcium gluconate, sodium gluconate, potassium gluconate, magnesium gluconate, zinc citrate, calcium citrate, sodium citrate, potassium citrate and magnesium citrate.

[0015] Preferably, the concentration of the metal salt solution is: 10 -3 ~10 -2 %wt.

[0016] Preferably, the mass ratio of Tremella polysaccharide, metal salt and fullerenol is: 476.0~498.8:1.0~20.0:0.2~4.0.

[0017] Preferably, the mass ratio of Tremella polysaccharide, metal salt and fullerenol is: 491.0:8.5:0.5.

[0018] Preferably, the reaction endpoint is the complete disappearance of the Tremella polysaccharide in the suspension.

[0019] Preferably, the heating temperature in step 4 is 45-55° C., and the ultrasonic condition is 25-35 min.

[0020] Preferably, the post-drying treatment method in step 4 is one of freeze drying, spray drying and forced air drying.

[0021] The second technical solution adopted by the present invention is: a copolymer of Tremella polysaccharide grafted with metal ions and fulleranol.

[0022] The third technical solution adopted by the present invention is: application of a copolymer of Tremella fuciformis polysaccharide grafted with metal ions and fullerol in cosmetics.

[0023] Beneficial effects of the above technical solution:

[0024] (1) Compared with the prior art, the Tremella polysaccharide grafted metal ion and fullerol copolymer provided by the present invention can meet the requirements of cosmetics in terms of moisturizing, anti-inflammatory, repairing, anti-oxidation and other aspects. The raw material used is fullerol, which does not contain fullerene nanomaterials, thus solving the safety and hydrophobicity problems of fullerene nanomaterials used in cosmetics, and has the ability to scavenge free radicals and protect skin cells from damage caused by the external environment such as ultraviolet rays, pollutants, etc.

[0025] (2) The preparation method provided by the present invention includes compounding metal ions such as calcium ions and zinc ions with Tremella polysaccharide, grafting and copolymerizing with fullerol, forming Tremella polysaccharide grafted metal ions and fullerol copolymers, improving the anti-inflammatory properties of the raw materials, solving the problem of difficulty in grafting fullerene with Tremella polysaccharide in the prior art, and ensuring that fullerol is uniformly grafted on the Tremella polysaccharide compound. The antioxidant property of the product is improved, and free radicals are effectively removed. In the present invention, the amount of fullerol added can be strictly controlled to ensure the stability, safety and efficacy of the material.

[0026] (3) The graft copolymer provided by the present invention is used in cosmetics, and the efficacy and performance are tested. The results show that it has the comprehensive efficacy of high-efficiency moisturizing, anti-inflammatory, repairing and anti-oxidation, and can meet the user's demand for the multi-faceted efficacy of cosmetics. At the same time, the raw material has good stability and safety. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 The present invention provides a flow chart of the preparation method of the Tremella polysaccharide grafted metal ion and fulleranol copolymer. DETAILED DESCRIPTION

[0028] The following is a further detailed description of the implementation methods of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than an exhaustive list of all the embodiments. It should be noted that the embodiments and features in the embodiments of the present application can be combined with each other without conflict.

[0029] The terms "first", "second", etc. (if any) in the specification and claims are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence. It should be understood that the terms used in this way are interchangeable where appropriate so that the embodiments described herein can be implemented in an order other than that illustrated or described herein. In addition, the terms "including" and "having" and any variations thereof are intended to cover non-exclusive inclusions, for example, a process, method, system, product, or device that includes a series of steps or units is not necessarily limited to those steps or units that are clearly listed, but may include other steps or units that are not clearly listed or inherent to these processes, methods, products, or devices.

[0030] It should be understood that the term "and / or" used in this article is only a description of the association relationship of associated objects, indicating that there can be three relationships. For example, A and / or B can represent: A exists alone, A and B exist at the same time, and B exists alone. In addition, the character " / " in this article generally indicates that the associated objects before and after are in an "or" relationship.

[0031] Raw materials and equipment (all commercially available)

[0032] Raw materials: Tremella polysaccharide, fullerol and metal salts, the metal salts include zinc gluconate, calcium gluconate, sodium gluconate, potassium gluconate, magnesium gluconate, one or more of zinc citrate, calcium citrate, sodium citrate, potassium citrate and magnesium citrate.

[0033] Equipment: vortex mixer, dryer (freeze dryer, spray dryer, air dryer are all acceptable);

[0034] The preparation method of Tremella polysaccharide grafted metal ions and fullerol copolymer comprises the following steps:

[0035] Step 1: Weigh raw materials, including metal salts, Tremella polysaccharide and fullerol; the metal salts include one or more of zinc gluconate, calcium gluconate, sodium gluconate, potassium gluconate, magnesium gluconate, zinc citrate, calcium citrate, sodium citrate, potassium citrate and magnesium citrate.

[0036] Step 2: Dissolve the metal salt in deionized water to prepare a metal salt solution; the concentration of the metal salt solution is: 10 -3 ~10 -2 %wt.

[0037] Step 3: Add the Tremella polysaccharide and fullerol to the metal salt solution to obtain a suspension, which is a visible Tremella polysaccharide suspension. The mass ratio of Tremella polysaccharide, metal salt, and fullerol is: 476.0~498.8: 1.0~20.0: 0.2~4.0.

[0038] Step 4: heating the suspension at 45-55°C, reacting under ultrasonic conditions for 25-35 minutes until the end of the reaction, and completely eliminating the Tremella polysaccharide in the suspension. Rotary evaporation to 1 / 3 volume, cooling, and drying to obtain Tremella polysaccharide grafted metal ions and fullerene alcohol copolymers. The drying post-treatment method is one of freeze drying, spray drying, and forced air drying.

[0039] Preferably, since the metal salt includes one or more of zinc gluconate, calcium gluconate, sodium gluconate, potassium gluconate, magnesium gluconate, zinc citrate, calcium citrate, sodium citrate, potassium citrate and magnesium citrate, the metal salt can provide corresponding metal ions. The following examples only take some metal salts as examples to prepare copolymers and test corresponding properties.

[0040] The chemical structure of Tremella polysaccharide is as follows:

[0041] ;

[0042] Tremella polysaccharide undergoes esterification reaction with fullerol and is connected with metal ions. The specific structural formula is shown below:

[0043]

[0044] Example 1

[0045] (1) Weigh 8.5 mg of zinc gluconate and prepare 24.5 mL of solution.

[0046] (2) Weigh 491.0 mg of Tremella polysaccharide and 0.5 mg of fullerol and dissolve them in 24.5 mL of metal salt solution.

[0047] (3) The mixed solution was shaken on a vortex mixer, heated to 45-55°C, and then sonicated for 30 min.

[0048] (4) Tremella polysaccharide is completely dissolved, cooled, and becomes gel-like.

[0049] (5) The gel is placed in a freeze dryer and dried to obtain copolymer 1.

[0050] Example 2

[0051] (1) Weigh 20.0 mg of zinc gluconate and prepare 24.5 mL of solution.

[0052] (2) Weigh 478.0 mg of Tremella polysaccharide and 2.0 mg of fullerenol and dissolve them in 24.5 mL of metal salt solution.

[0053] (3) Place on a vortex mixer, shake well, heat to 45-55°C, and then sonicate for 30 min.

[0054] (4) Tremella polysaccharide is completely dissolved, cooled, and becomes gel-like.

[0055] (5) The gel is placed in a spray dryer and dried to obtain copolymer 2.

[0056] Example 3

[0057] (1) Weigh 1.0 mg of zinc gluconate and prepare 24.5 mL of solution.

[0058] (2) Weigh 498.9 mg of Tremella polysaccharide and 0.1 mg of fullerol and dissolve them in 24.5 mL of metal salt solution.

[0059] (3) Place on a vortex mixer, shake well, heat to 45-55°C, and then sonicate for 30 min.

[0060] (4) Tremella polysaccharide is completely dissolved, cooled, and becomes gel-like.

[0061] (5) The gel is placed in an air dryer and dried to obtain copolymer 3.

[0062] Example 4

[0063] (1) Weigh 1.0 mg of zinc gluconate and prepare 24.5 mL of solution.

[0064] (2) Weigh 497.0 mg of Tremella polysaccharide and 2.0 mg of fullerol and dissolve them in 24.5 mL of metal salt solution.

[0065] (3) Place on a vortex mixer, shake well, heat to 45-55°C, and then sonicate for 30 min.

[0066] (4) Tremella polysaccharide is completely dissolved, cooled, and becomes gel-like.

[0067] (5) The gel was placed in a freeze dryer and dried to obtain copolymer 4.

[0068] Example 5

[0069] (1) Weigh 20.0 mg of zinc gluconate and prepare 24.5 mL of solution.

[0070] (2) Weigh 479.9.0 mg of Tremella polysaccharide and 0.1 mg of fullerol and dissolve them in 24.5 mL of metal salt solution.

[0071] (3) Place on a vortex mixer, shake well, heat to 45-55°C, and then sonicate for 30 min.

[0072] (4) Tremella polysaccharide is completely dissolved, cooled, and becomes gel-like.

[0073] (5) The gel was placed in a freeze dryer and dried to obtain copolymer 5.

[0074] Example 6

[0075] (1) Weigh 13.4 mg of calcium gluconate and prepare 24.5 mL of solution.

[0076] (2) Weigh 486.1 mg of Tremella polysaccharide and 0.5 mg of fullerol and dissolve them in 24.5 mL of metal salt solution.

[0077] (3) Place on a vortex mixer, shake well, heat to 45-55°C, and then sonicate for 30 min.

[0078] (4) Tremella polysaccharide is completely dissolved, cooled, and becomes gel-like.

[0079] (5) The gel was placed in a freeze dryer and dried to obtain copolymer VI.

[0080] Example 7

[0081] (1) Weigh 20.0 mg of calcium gluconate and prepare 24.5 mL of solution.

[0082] (2) Weigh 478.0 mg of Tremella polysaccharide and 2.0 mg of fullerenol and dissolve them in 24.5 mL of metal salt solution.

[0083] (3) Place on a vortex mixer, shake well, heat to 45-55°C, and then sonicate for 30 min.

[0084] (4) Tremella polysaccharide is completely dissolved, cooled, and becomes gel-like.

[0085] (5) The gel was placed in a freeze dryer and dried to obtain copolymer VII.

[0086] Example 8

[0087] (1) Weigh 20.0 mg of calcium gluconate and prepare 24.5 mL of solution.

[0088] (2) Weigh 498.9 mg of Tremella polysaccharide and 2.0 mg of fullerenol and dissolve them in 24.5 mL of metal salt solution.

[0089] (3) Place on a vortex mixer, shake well, heat to 45-55°C, and then sonicate for 30 min.

[0090] (4) Tremella polysaccharide is completely dissolved, cooled, and becomes gel-like.

[0091] (5) The gel is placed in an air dryer and dried to obtain copolymer VIII.

[0092] Example 9

[0093] (1) Weigh 1.0 mg of calcium gluconate and prepare 24.5 mL of solution.

[0094] (2) Weigh 497.0 mg of Tremella polysaccharide and 2.0 mg of fullerol and dissolve them in 24.5 mL of metal salt solution.

[0095] (3) Place on a vortex mixer, shake well, heat to 45-55°C, and then sonicate for 30 min.

[0096] (4) Tremella polysaccharide is completely dissolved, cooled, and becomes gel-like.

[0097] (5) The gel was placed in an air dryer and dried to obtain copolymer 9.

[0098] Example 10

[0099] (1) Weigh 20.0 mg of calcium gluconate and prepare 24.5 mL of solution.

[0100] (2) Weigh 479.9 mg of Tremella polysaccharide and 0.1 mg of fullerol and dissolve them in 24.5 mL of metal salt solution.

[0101] (3) Place on a vortex mixer, shake well, heat to 45-55°C, and then sonicate for 30 min.

[0102] (4) Tremella polysaccharide is completely dissolved, cooled, and becomes gel-like.

[0103] (5) The gel is placed in an air drying machine for drying to obtain copolymer 10.

[0104] The components and their contents in Examples 1 to 10 are shown in Table 1.

[0105] Table 1 Components and contents in each example

[0106]

[0107] Comparative Example 1

[0108] A water-soluble fullerene complex in the prior art is used as comparative example 1. The preparation method of the water-soluble fullerene complex in comparative example 1 is as follows:

[0109] (1) mixing fullerene powder and Tremella polysaccharide evenly;

[0110] (2) subjecting the mixture of the fullerene powder and Tremella polysaccharide to a strong dispersion treatment to obtain a mixture powder material;

[0111] (3) adding water to fully dissolve the obtained powder material, and centrifuging at high speed in a centrifugal device to obtain a supernatant;

[0112] (4) Filtering the supernatant using a filtering device, concentrating and drying the supernatant to obtain the water-soluble fullerene complex.

[0113] Comparative Example 2

[0114] A moisturizing dry facial mask containing fullerene and a preparation method thereof in the prior art is used as comparative example 2. The preparation method of the water-soluble fullerene complex of comparative example 2 is as follows:

[0115] 0.08 parts of palmitoyl tripeptide-8, 0.1 parts of okra polysaccharide, 0.2 parts of hydrolyzed sclerotium gum, 0.25 parts of fullerene, 0.3 parts of tremella polysaccharide, 0.5 parts of β-glucan, 0.5 parts of D-panthenol, 0.6 parts of tocopherol, 0.8 parts of Astragalus membranaceus root extract, 1 part of silk peptide, 1.2 parts of aloe barbadensis leaf extract, 2 parts of mannitol, 3.5 parts of 1,3-butylene glycol, 4 parts of moisturizing composition, 5 parts of glycerin, and 79.97 parts of deionized water are stirred and mixed evenly.

[0116] Comparative Example 3

[0117] Taking the Tremella polysaccharide solution in the prior art as comparative example 3, the preparation method is as follows:

[0118] (1) Weigh 500.0 mg of Tremella polysaccharide and dissolve it in 24.5 mL of deionized water;

[0119] (2) Place the mixture on a vortex mixer and shake well, then sonicate for 30 min;

[0120] (3) Tremella polysaccharide was completely dissolved and became gel-like after cooling;

[0121] (4) Place the gel in a freeze dryer for freeze drying and take it out after freeze drying.

[0122] Performance detection and evaluation method of Tremella polysaccharide grafted with metal ions and fulleranol copolymer

[0123] 1 Evaluation of antioxidant capacity index ORAC value

[0124] Accurately pipette 1g of sample into a 15ml centrifuge tube and dilute with 10ml ultrapure water to obtain a 100mg / ml sample solution. Through preliminary experiments, find the appropriate concentration for DPPH evaluation and finally obtain the concentration series of the sample:

[0125] The ORAC reaction is carried out in a 75 mmol potassium phosphate buffer (pH = 7.4) environment. After pre-setting at 37°C for 5 minutes, 140 μL of AAPH is quickly added to each well with a multichannel pipette to start the reaction. The microplate is placed in a fluorescence analyzer at 37°C for continuous measurement with an excitation wavelength of 485 nm and an emission wavelength of 538 nm. The fluorescence intensity of each well is measured every 2 minutes, and the measurement time is generally set until the fluorescence decays to the baseline.

[0126] 2 Safety evaluation

[0127] According to the "Technical Specifications for Safety of Cosmetics" (2015 edition), with clinical medicine as the theoretical basis, a specific experimental population is formed into a test group, and a patch test is conducted on the designated parts of the subjects to determine the potential possibility of adverse reactions of cosmetics to human skin. Select qualified patch test materials. Put the test substance into the patch tester, with an amount of about 0.020g~0.025g (solid or semi-solid) or 0.020mL~0.025mL (liquid, which can be dripped on the filter paper attached to the patch tester and placed in the patch tester).

[0128] When the test substance is the original cosmetic final product, the control well is a blank control (no substance is placed in it). When the test substance is a diluted cosmetic, the diluent of the cosmetic is used in the control well.

[0129] Apply the patch tester containing the test substance to the back or curved forearm of the subject with non-irritating tape, and press lightly with the palm of your hand to make it evenly adhere to the skin for 24 hours.

[0130] Observe the skin reaction 30 minutes after removing the test patch tester and after the indentation disappears. If the result is negative, observe again 24 hours and 48 hours after the patch test. Record the reaction results according to Table 2 (Grading Standards for Adverse Skin Reactions).

[0131] Table 2 Grading table for adverse skin reactions

[0132]

[0133] Interpretation of skin occlusive patch test results: If more than 5 out of 30 subjects experience Grade 1 adverse skin reactions, or more than 2 subjects experience Grade 2 adverse skin reactions, or any Grade 3 or above adverse skin reaction occurs, the test substance is judged to have an adverse reaction to the human body.

[0134] 3Anti-aging effect evaluation

[0135] Studies have shown that senescent cells have significant characteristics. Senescent cells usually become larger in size, and the activity of senescence-associated β-galactosidase (SA-β-Gal) is upregulated in a pH 6.0 environment. Therefore, the anti-aging effect of the active ingredient was evaluated through the β-galactosidase activity inhibition experiment. The evaluation results are shown in Table 3.

[0136] Table 3 Anti-aging efficacy test and evaluation results

[0137]

[0138] From the detection, evaluation and analysis of different embodiments in Table 3 above, it can be concluded that the Tremella polysaccharide grafted metal ions and fullerol copolymers provided by the present invention have better antioxidant properties than the grafted copolymers provided by the comparative example. The Tremella polysaccharide grafted metal ions and fullerol copolymers provided by Examples 1 and 6 have outstanding antioxidant properties, and Example 1 has better antioxidant properties and superior anti-aging ability.

[0139] 4 Evaluation of anti-inflammatory efficacy

[0140] 4.1 Cell survival rate

[0141] Add 10 μL of WST-8 staining solution in the dark, incubate at 37°C for 30 min, and use an enzyme-linked immunosorbent assay to measure the absorbance at 450 nm. For the blank control, mix an equal amount of cell-free culture medium and WST-8 staining solution to calculate the cell survival rate.

[0142] 4.2 Antioxidant damage repair

[0143] The level of intracellular reactive oxygen species (ROS) in human dermal fibroblasts was determined by the DCFH-DA method. After culturing the cells under UV irradiation for 24 h, each group of cells was washed twice with PBS, incubated with DCFH-DA for 30 min, digested with trypsin, centrifuged and collected. The cells were resuspended, observed with a fluorescence microscope, and the fluorescence intensity was measured.

[0144] 4.3 Anti-inflammatory effect

[0145] Ultraviolet radiation can lead to an increase in the levels of inflammatory factors IL-1B, IL-6 and TNF-a in skin tissue, while treatment with the composition of the present invention can reduce the abnormal levels of inflammatory factors in skin tissue caused by ultraviolet radiation.

[0146] Blank group: Cultured in untreated DMEM medium.

[0147] Model group: The fibroblasts were cultured in untreated DMEM medium and irradiated with UVA and UVB.

[0148] Experimental group: The fibroblasts were cultured with a culture medium containing the copolymers of different embodiments, and were irradiated with UVA and UVB.

[0149] Each of the above groups of cells was set up with 5 replicate wells. The amount of the composition added to each group was the same, all 100 μL. The anti-inflammatory efficacy test evaluation results are shown in Table 4.

[0150] Table 4 Anti-inflammatory efficacy test evaluation results

[0151]

[0152] From the detection and evaluation analysis of different embodiments in Table 4 above, compared with the blank group and the model group, it can be concluded that the experimental results of the Tremella polysaccharide grafted metal ions and fullerol copolymers of Examples 1 to 10 have better cell survival rate, antioxidant damage repair and anti-inflammatory effects, while the compositions of Comparative Examples 1 to 3 lack a certain component of the present invention and have poor effects. It can be seen that the components in the Tremella polysaccharide grafted metal ions and fullerol copolymers provided by the present invention have a synergistic effect, which can improve cell survival rate, repair oxidative damage and have anti-inflammatory effects.

[0153] The Tremella polysaccharide grafted metal ions and fullerol copolymer provided in Example 1 has better antioxidant properties than other groups, and the Tremella polysaccharide grafted metal ions and fullerol copolymer provided in Example 1 and Example 6 have better anti-inflammatory and superior antioxidant damage repair capabilities.

[0154] 5. Propionibacterium acnes

[0155] Fluorescence spectroscopy: Porphyrins in Propionibacterium acnes emit fluorescence at specific wavelengths. The presence and quantity of porphyrins can be determined by irradiating the sample with an excitation light source (such as a laser or LED) and detecting the emitted fluorescence using a spectrometer. This method has a high degree of specificity and sensitivity. The test results of the Tremella polysaccharide grafted metal ions and fullerol copolymers provided in preferred embodiments 1 and 6 are shown in Table 5 below.

[0156] Table 5 Porphyrin quantity detection results

[0157]

[0158] 6Inhibit tyrosinase activity

[0159] In the biosynthesis of skin melanin, tyrosinase is a key enzyme that acts on DOPA to form DOPA quinone, which spontaneously undergoes a series of reactions to finally form melanin. In a phosphoric acid solution at pH 6.8, tyrosinase can catalyze the conversion of DOPA into DOPA quinone, and the absorbance value can be measured at 475nm on a spectrophotometer. Raw materials with tyrosinase activity inhibition can reduce the conversion of DOPA into DOPA quinone, thereby reducing the absorbance value. The inhibitory effect of the raw materials on tyrosinase activity can be evaluated based on the change in absorbance value.

[0160] Test results

[0161] Each batch of experiments needs to be tested with a positive control. The IC50 (the sample test concentration corresponding to 50% inhibition effect) of the positive control kojic acid should be between 0.05 mg / mL and 0.15 mg / mL, and the test system is considered to be effective. The samples provided in Examples 1 to 10 and Comparative Examples 1 to 3 were tested, and the results are shown in Tables 6 and 7 below.

[0162] Table 6 Tyrosinase inhibition test results 1

[0163]

[0164] Table 7 Tyrosinase inhibition test results II

[0165]

[0166] Compared with the raw materials provided in Examples 1 to 10 and Comparative Examples 1 to 3, the Tremella polysaccharide grafted metal ion and fulleranol copolymer provided in Example 1 has better tyrosine inhibition ability.

[0167] According to the above performance test evaluation, the tremella polysaccharide grafted metal ions and fullerol copolymers provided by the present invention have significantly improved performance compared with the raw materials in the prior art: in the cosmetics industry, seeking efficient and multifunctional raw materials has always been the focus of research and development. The tremella polysaccharide grafted metal ions and fullerol copolymers provided by the present invention not only maintain the excellent moisturizing, anti-inflammatory and repairing properties of tremella polysaccharides, but also significantly improve their anti-inflammatory effects by adding metal ions (including Ca / Zn ions). At the same time, the introduction of fullerol adds a strong antioxidant capacity to the product, effectively resisting the damage of the external environment to the skin. This comprehensive performance improvement gives the tremella polysaccharide grafted metal ions and fullerol copolymers a unique competitive advantage in the field of cosmetics.

[0168] Proper cost control: Although the research and development process of the Tremella polysaccharide grafted metal ions and fullerol copolymer involves multiple ingredients, the cost-effectiveness of the raw materials is ensured through meticulous cost control. While maintaining product performance, the production cost is reduced to provide a more reasonable market price to meet consumer demand.

[0169] Easy to process and apply: The Tremella polysaccharide grafted metal ions and fullerol copolymer has good stability and compatibility, and is easy to mix and process with other cosmetic ingredients. It provides cosmetic manufacturers with greater formula design space and simplifies the production process. In addition, the raw material is easy to store and transport, reducing the risk in the production process.

[0170] Environmental protection and sustainability: The preparation process of the Tremella polysaccharide grafted metal ions and fullerol copolymer uses environmentally friendly materials and processes, reducing pollution to the environment. At the same time, the raw material has good biodegradability and will not cause long-term burden on the environment. It is in line with the green development concept of modern society and has also contributed to the sustainable development of the cosmetics industry.

[0171] High safety: Safety is an important consideration in the cosmetics industry. The Tremella polysaccharide grafted metal ions and fullerol copolymer of the present invention has undergone strict safety assessment and quality control during the preparation process. It is ensured that the raw materials used comply with relevant regulations and standards and are non-irritating to the skin. This highly safe raw material provides consumers with a safer and more reliable skin care experience.

[0172] The present invention not only retains the natural moisturizing, anti-inflammatory and repairing properties of Tremella polysaccharide through a unique combination of raw materials and graft copolymerization technology, but also significantly enhances its anti-inflammatory and antioxidant capabilities. This comprehensive functional improvement makes the present invention have a broader application prospect in the field of cosmetics. Combining Tremella polysaccharide and metal ions, not only can their respective functions be fully exerted, but also the skin care effect is significantly enhanced through synergy. Compared with single-ingredient products, it more comprehensively meets the multiple needs of modern people for cosmetics, integrates anti-oxidation, anti-inflammatory, moisturizing and soothing, and realizes comprehensive skin care. At the same time, by optimizing the production process and ensuring the stability of the formula. Fullerol is compounded with the Ca / Zn ion Tremella polysaccharide composite gel through an esterification reaction, ensuring that fullerol can be evenly grafted on Tremella polysaccharide, thereby giving full play to its antioxidant effect. This step is the key to the comprehensive functional improvement of the present invention, so that the graft copolymer provided by the present invention has higher moisturizing, anti-inflammatory, repairing, antioxidant and protection against ultraviolet damage.

[0173] When the concentration of Tremella polysaccharide is 10 mg / mL or above, the solution can form a gel, and the viscosity of the gel increases with the increase of Tremella polysaccharide concentration. The concentration of metal ions should not be too high, and high concentration of metal ions will reduce the viscosity of the gel, which will comprehensively affect the function and activity of the final product.

[0174] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not limitations on the implementation methods of the present invention. For ordinary technicians in the relevant field, other different forms of changes or modifications can be made on the basis of the above description. It is impossible to list all the implementation methods here. All obvious changes or modifications derived from the technical solution of the present invention are still within the protection scope of the present invention.

Claims

1. A method for preparing a copolymer of Tremella fuciformis polysaccharide grafted with metal ions and fullerol, characterized in that: include: Step 1: weighing raw materials, including metal salt, Tremella polysaccharide and fullerenol; Step 2: dissolving the metal salt in deionized water to prepare a metal salt solution; Step 3: adding the Tremella polysaccharide and fullerenol to the metal salt solution to obtain a suspension; Step 4: heating the suspension, reacting to the end point of the reaction under ultrasonic conditions, rotary evaporating to 1 / 3 of the liquid volume, cooling, drying and post-processing to obtain the Tremella polysaccharide grafted metal ion and fullerenol copolymer; The metal salt includes one or more of zinc gluconate, calcium gluconate, zinc citrate and calcium citrate.

2. The method for preparing the copolymer of Tremella fuciformis polysaccharide grafted with metal ions and fullerol according to claim 1, characterized in that: The mass concentration of the metal salt solution is: 10 -3 ~10 -2 %wt.

3. The method for preparing the copolymer of Tremella fuciformis polysaccharide grafted with metal ions and fullerol according to claim 1, characterized in that: The mass ratio of the Tremella polysaccharide, the metal salt and the fullerenol is: 476.0-498.8:1.0-20.0:0.2-4.

0.

4. The method for preparing the copolymer of Tremella fuciformis polysaccharide grafted with metal ions and fullerol according to claim 1, characterized in that: The mass ratio of Tremella polysaccharide, metal salt and fullerenol is 497.5:8.5:

2.

5. The method for preparing the copolymer of Tremella fuciformis polysaccharide grafted with metal ions and fullerol according to claim 1, characterized in that: The reaction endpoint is when the Tremella polysaccharide in the suspension completely disappears.

6. The method for preparing the copolymer of Tremella fuciformis polysaccharide grafted with metal ions and fullerol according to claim 1, characterized in that: The heating temperature in step 4 is 45-55° C., and the ultrasonic condition is 25-35 min.

7. The method for preparing the copolymer of Tremella fuciformis polysaccharide grafted with metal ions and fullerol according to claim 1, characterized in that: The post-drying treatment method in step 4 is one of freeze drying, spray drying and air drying.

8. A copolymer of Tremella polysaccharide grafted with metal ions and fullerol prepared by the preparation method according to any one of claims 1 to 7.

9. Use of the copolymer of Tremella fuciformis polysaccharide grafted with metal ions and fullerol as claimed in claim 8 in cosmetics.

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