Ganoderma lucidum spore polysaccharide as well as preparation method and application thereof

Abstract

The invention relates to the technical field of ganoderma lucidum polysaccharide extraction, and particularly discloses ganoderma lucidum spore polysaccharide as well as a preparation method and application thereof. A preparation method of ganoderma lucidum spore polysaccharide comprises the following steps: washing ganoderma lucidum spore powder with water, removing impurities, drying in vacuum, embrittling at-19 to-22 DEG C, and then carrying out ultrasonic mixing and wall breaking to obtain pretreated ganoderma lucidum spore powder; adding the pretreated ganoderma lucidum spore powder into water, mixing and stirring, performing ultrasonic extraction, standing, and centrifuging to obtain a first extracting solution; adjusting the pH value of the first extracting solution, adding trypsin for enzymolysis, centrifuging, adding cation exchange resin, stirring for enzyme removal, filtering, concentrating to 1 / 3 of the original volume, and performing vacuum freeze drying to obtain the product. The ganoderma lucidum spore polysaccharide disclosed by the invention can be used for preparing the periorbital fat granule removing gel, and has the advantages of eliminating periorbital fat granules, improving periorbital dark circles and under-eye puffiness and brightening skin color.

Description

Technical Field

[0001] The present application relates to the technical field of Ganoderma lucidum polysaccharide extraction, and more specifically, it relates to a Ganoderma lucidum spore polysaccharide, its preparation method and application. Background Art

[0002] Ganoderma lucidum is a medicinal fungus belonging to the genus Ganoderma in the family Polyporaceae, order Polyporales, class Basidiomycetes. It has aliases such as right ear and is also known as wood Ganoderma lucidum, mushroom Ganoderma lucidum, soul grass, fairy grass, etc. It is widely distributed in tropical and subtropical regions. It tastes sweet and is neutral in nature, and belongs to the heart, liver, lung, and kidney meridians, with the effects of nourishing the heart and calming the mind, relieving asthma and coughing, tonifying the lung and replenishing qi, etc. Modern research shows that Ganoderma lucidum contains various bioactive components such as nucleotides, triterpenoids, polysaccharides, glycoproteins, and alkaloids. Among them, Ganoderma lucidum spore polysaccharide has shown great application potential in the fields of food, medicine, and cosmetics due to its multiple pharmacological activities such as immunomodulation, anti-tumor, hypoglycemic, anti-inflammatory, and anti-aging.

[0003] Ganoderma lucidum spore polysaccharide is the main bioactive component extracted and separated from Ganoderma lucidum spore powder. Small molecular polysaccharides can penetrate the cell wall and be directly absorbed by cells, participating in the metabolic regulation activities of cells, which provides a broad research prospect for the preparation of functional skin care products in the cosmetics field.

[0004] Due to the complexity of Ganoderma lucidum spore polysaccharide, the structure and properties of Ganoderma lucidum spore polysaccharide are affected by various aspects such as the origin, variety, and extraction method of Ganoderma lucidum. The quality of the Ganoderma lucidum spore polysaccharide preparation technology directly affects the quality of Ganoderma lucidum spore polysaccharide. Selecting a suitable preparation method is crucial for the quality and application of Ganoderma lucidum spore polysaccharide. In view of the related technologies mentioned above, the inventor found that traditional preparation methods such as hot water extraction often have the problem of low extraction efficiency, while microwave-assisted method and ultrasonic extraction method, although having high extraction efficiency, may damage the structure of polysaccharides, thus affecting their biological activity. Summary of the Invention

[0005] In order to improve the preparation efficiency of Ganoderma lucidum spore polysaccharide and maintain the biological activity of Ganoderma lucidum spore polysaccharide, and expand the application of Ganoderma lucidum spore polysaccharide, the present application provides a Ganoderma lucidum spore polysaccharide, its preparation method and application.

[0006] In the first aspect, the present application provides a preparation method of Ganoderma lucidum spore polysaccharide, adopting the following technical scheme: A preparation method of Ganoderma lucidum spore polysaccharide, comprising the following steps: S1: Wash, remove impurities, and vacuum dry Ganoderma lucidum spore powder, then place it at -19 to -22 °C for embrittlement for 4.5 - 5.5 h, and then perform ultrasonic mixing and cell wall breaking at 45 - 50 °C for 3 - 5 min to obtain pretreated Ganoderma lucidum spore powder; S2: Add the pretreated Ganoderma lucidum spore powder into water at a material-liquid ratio of 1:(20 - 30), mix and stir for 20 - 30 min, then perform ultrasonic extraction for 5 - 8 min, and then let it stand for 5 - 6 h, and centrifuge to obtain the first extract; S3: Adjust the pH of the first extract to (8 ± 0.2), add trypsin, and perform water bath heating and enzymatic hydrolysis at 35 - 45 °C for 1 - 1.5 h, and then centrifuge to obtain the second extract; S4: Add cation exchange resin to the second extract, stir and remove the enzyme for 1 - 1.5 h, and filter to obtain the third extract; S5: Concentrate the third extract to 1 / 3 of its original volume under vacuum, and then perform vacuum freeze-drying to obtain the product.

[0007] By adopting the above technical solutions, the technological process of preparing Ganoderma lucidum spore polysaccharide is carried out under low-temperature conditions, which can effectively retain the biological activity of the polysaccharide in the broken Ganoderma lucidum spore powder and protect it from the influence of temperature factors.

[0008] Through the pretreatment of freeze-crystallization embrittlement and ultrasonic mixing and cell wall breaking of the Ganoderma lucidum spore powder after vacuum drying, the cell wall of the Ganoderma lucidum spore powder can be made more fragile first, which is convenient for efficiently destroying the cell wall of the Ganoderma lucidum spore powder under the strong cavitation effect and mechanical effect during ultrasonic treatment, so that its polysaccharide and other active ingredients can be more easily released during the subsequent water-soluble process, effectively improving the extraction efficiency and quality of the subsequent Ganoderma lucidum spore polysaccharide.

[0009] After the pretreated Ganoderma lucidum spore powder is dissolved in water, the extraction of Ganoderma lucidum spore polysaccharide is carried out by means of preliminary stirring, short-term ultrasonic treatment and standing, which just makes the dissolution efficiency of Ganoderma lucidum spore polysaccharide reach the best level, prevents the continuous ultrasonic from over-damaging the active ingredients, and improves the extraction rate of polysaccharide by combining with the mass transfer efficiency of the solution.

[0010] Protein belongs to macromolecular substances, and its direct utilization rate is relatively low. It is not easy to penetrate the skin to play a direct role when preparing cosmetics. Trypsin treatment can hydrolyze the macromolecular proteins in the Ganoderma lucidum spore powder in the first extract and degrade them into small peptides and amino acids. On the one hand, because small peptides and amino acids have small molecular weights and good skin permeability, they can carry Ganoderma lucidum spore polysaccharide into the deep layer of the skin more effectively, so as to play better biological activity. On the other hand, small peptides and amino acids can act as auxiliary components to cooperate with Ganoderma lucidum spore polysaccharide and expand its application in the preparation of functional cosmetics.

[0011] Optionally, the addition amount of the trypsin is 0.1 - 0.2% of the mass of the first extract, and the enzyme activity is 200,000 u / g.

[0012] By adopting the above technical solution and controlling the dosage of trypsin, the protein in the first extract can be efficiently enzymolyzed while avoiding the destruction of Ganoderma lucidum spore polysaccharide, thereby improving the purity and biological activity of Ganoderma lucidum spore polysaccharide.

[0013] Optionally, the cation exchange resin is a macroporous resin with a sulfide group, and the addition amount is 15-20 g / L.

[0014] By adopting the above technical solution and using a macroporous structure resin with a sulfonic acid group, trypsin can be efficiently removed and the purity of Ganoderma lucidum spore polysaccharide can be improved.

[0015] Optionally, the frequency of ultrasonic mixing and wall breaking in step S1 is 20-30kHz, the power is 300-500W, and the frequency of ultrasonic extraction in step S2 is 30-40kHz, the power is 600-800W, and the temperature is 45-55°C.

[0016] In a second aspect, the present application provides a Ganoderma lucidum spore polysaccharide, which is prepared by the Ganoderma lucidum spore polysaccharide preparation method of the present application.

[0017] In a third aspect, the present application provides a use of Ganoderma lucidum spore polysaccharide in the preparation of a gel for removing fat particles around the eyes.

[0018] In a fourth aspect, the present application provides a gel for removing fat particles around the eyes.

[0019] A gel for removing fat particles around the eyes, wherein the raw materials include, by mass percentage, 6-8% of the ganoderma lucidum spore polysaccharide according to claim 5, 1-3% fruit acid, 0.4-0.7% salicylic acid, 0.5-1% p-hydroxyacetophenone, 5-10% glycerol, 2-5% squalane, 2-5% emulsifier, 0.5-1.5% xylitol glucoside, 1-2% anhydroxylitol, 1-2% vitamin B5, 2-5% avocado tree fat, 0.5-1.5% sorbitan isostearate and 4-6% emollient components, and the balance is water.

[0020] Fat particles are usually tiny cysts formed by abnormal skin keratinization and sebum accumulation. Fruit acid and salicylic acid can promote the metabolism of keratinocytes, penetrate into lipid pores, remove acne dirt and excess oil accumulated in the pores to dissolve the accumulated keratin and sebum, thereby effectively reducing and removing fat particles. However, since the skin around the eyes is relatively weak and sensitive, the use of acid ingredients may cause skin rejection reactions, such as peeling, redness, swelling and inflammation, etc. These adverse reactions may increase the burden on the skin around the eyes.

[0021] By adopting the above technical solutions, the Ganoderma lucidum spore polysaccharide of the present application can be used in combination with fruit acid and salicylic acid as a skin care product for the eye area, effectively neutralizing the skin rejection reaction that may be caused by acid components, further effectively caring for the skin around the eyes, removing dead skin cells and excess oil, and promoting the removal of milia. In addition, the Ganoderma lucidum spore polysaccharide of the present application also helps to reduce dark circles and eye bags when preparing skin care products for the eye area, and improves the overall condition of the skin around the eyes.

[0022] Optionally, the fruit acid is glycolic acid and citric acid with a mass ratio of 1:(0.4 - 0.6).

[0023] By adopting the above technical solutions, glycolic acid and citric acid, as a kind of fruit acid, have high solubility, can effectively soften the skin surface layer, remove the dead skin epidermis, promote the keratin metabolism of the skin around the eyes, dissolve sebum accumulation, and effectively reduce and eliminate milia around the eyes.

[0024] Optionally, the emulsifier is lecithin and polysorbate - 60 with a mass ratio of 1:(2.5 - 4.5).

[0025] By adopting the above technical solutions, lecithin is one of the main components of cell membranes and has a similar phospholipid bilayer structure to skin cells, which can promote the interaction between active ingredients and skin cell membranes, ensuring that the active ingredients penetrate into the skin layer. And polysorbate - 60, as a non - ionic surfactant, can make the encapsulated active ingredients form tiny emulsion droplets in the gel, making various active ingredients more evenly dispersed.

[0026] The two work together as an emulsifier to effectively encapsulate the active ingredients in the gel, improve the stability of the gel, make various active ingredients more evenly dispersed, continuously penetrate into the skin, and significantly improve the absorption rate of the active ingredients in the gel by the skin around the eyes.

[0027] Optionally, the emollient component is selected from any one or a combination of polydimethylsiloxane, isononyl isononanoate, isoamyl acetate, and caprylic / capric triglyceride.

[0028] By adopting the above technical solutions, components such as polydimethylsiloxane, isononyl isononanoate, isoamyl acetate, and caprylic / capric triglyceride are added to skin care products as emollient components. They can not only enhance the moisturizing effect of skin care products on the skin, but also help to form a protective layer on the skin surface, reduce the influence of the external environment on the skin, make the skin care products smoother and easier to apply during use, and provide long - term moisturizing and protection for the skin around the eyes.

[0029] In summary, the present application has the following beneficial effects: 1. The preparation process of Ganoderma lucidum spore polysaccharide in this application is carried out under low-temperature conditions. After the vacuum-dried Ganoderma lucidum spore powder is pretreated by cryo-brittle and ultrasonic wall-breaking, it is dissolved in water, and the Ganoderma lucidum spore polysaccharide is extracted by combining stirring, short-term ultrasound and static standing. The hard cell wall of Ganoderma lucidum spore powder is efficiently destroyed, and the dissolution rate of Ganoderma lucidum spore polysaccharide reaches the optimal level appropriately, reducing the excessive damage of ultrasound to the effective active ingredients in Ganoderma lucidum spore powder, and effectively improving the preparation efficiency and quality of Ganoderma lucidum spore polysaccharide.

[0030] 2. This application uses trypsin to treat the extract of Ganoderma lucidum spore powder, degrading the macromolecular proteins that are difficult to penetrate the skin and play a direct role in Ganoderma lucidum spore powder into small peptides and amino acids. On the one hand, because small peptides and amino acids have smaller molecular weights and good skin permeability, they can carry Ganoderma lucidum spore polysaccharide into the deep layer of the skin more effectively, thereby exerting better biological activity. On the other hand, small peptides and amino acids can act as auxiliary components to synergize with Ganoderma lucidum spore polysaccharide, expanding its application in the preparation of functional cosmetics.

[0031] 3. The Ganoderma lucidum spore polysaccharide of this application can be applied to the preparation of eye perimeter milia removal gel, which can effectively reduce the skin rejection reaction that may be caused by salicylic acid and peracid substances to the eye perimeter skin, effectively enhance the efficacy of the eye perimeter milia removal gel in reducing and eliminating eye perimeter milia, and further improve the overall condition of the eye perimeter skin. Detailed implementation manners

[0032] The following examples further illustrate this application in detail. Raw materials

[0033] Unless otherwise specified, the raw materials used in the examples, comparative examples, application examples and comparative application examples in this application are all commercially available cosmetic-grade products, specifically: Ganoderma lucidum spore powder, selected from the Ganoderma lucidum spore powder of Xianlingzhi collected from the Jinzhai Dabie Mountain Ganoderma lucidum base; Trypsin, selected from Xiasheng GDG-2009, with an enzyme activity of 200,000 u / g; Cation exchange resin, a macroporous structure resin with sulfonic groups, selected from Hubei Rishengchang New Material Technology Co., Ltd., D001 macroporous styrene-based cation exchange resin; Salicylic acid, glycolic acid and citric acid, all of cosmetic grade, with a purity ≥ 99%; Sorbitan isostearate, selected from Span 120 / Span 120 LQ-(RB); Shea butter, selected from Shanxi Shuohang Biotechnology Co., Ltd., SHSW-0327-007; Polydimethylsiloxane, selected from Victy Bio, WKQ-0034782; Isononyl isononanoate, selected from SEPPIC LANOL99; Isoamyl acetate, selected from WAK-CHEM, WKQ-0063210; Caprylic / capric triglyceride, selected from KLK GTCC 3575. Examples Example 1

[0034] A method for preparing Ganoderma lucidum spore polysaccharide, comprising the following steps: S1: Wash and remove impurities from Ganoderma lucidum spore powder, then perform vacuum drying. Set the vacuum degree to 0.9 MPa and the vacuum drying time to 6 h, so that the moisture content of the Ganoderma lucidum spore powder after vacuum drying is ≤ 6%. Then place it at -19 °C for embrittlement for 5.5 h, and then at 45 °C, with a frequency of 20 kHz and a power of 300 W, perform ultrasonic mixing and wall-breaking for 5 min to obtain pretreated Ganoderma lucidum spore powder; S2: Add the pretreated Ganoderma lucidum spore powder to water according to a solid-liquid ratio of 1:20, mix and stir for 20 min, then at 45 °C, with a frequency of 30 kHz and a power of 600 W, perform ultrasonic extraction for 8 min, then let it stand for 6 h and centrifuge to obtain the first extract; S3: Adjust the pH of the first extract to (8 ± 0.2), add trypsin, perform water bath heating and enzymatic hydrolysis at 45 °C for 1 h and then centrifuge to obtain the second extract. The addition amount of trypsin is 0.1% of the mass of the first extract, and the enzyme activity is 200,000 u / g; S4: Add cation exchange resin to the second extract, stir and remove the enzyme for 1 h, filter to obtain the third extract. The addition amount of cation exchange resin is 15 g / L; S5: Concentrate the third extract under vacuum to 1 / 3 of the original volume, then set the vacuum degree to -0.2 MPa for vacuum freeze-drying treatment. First pre-cool at -16 °C for 12 h, then freeze at -22 °C for 24 h, and finally heat up to 45 °C, with a sublimation time of 20 h to obtain the product. Example 2

[0035] A method for preparing Ganoderma lucidum spore polysaccharide, comprising the following steps: S1: Wash and remove impurities from Ganoderma lucidum spore powder, then perform vacuum drying. Set the vacuum degree to 0.9 MPa and the vacuum drying time to 6 h, so that the moisture content of the Ganoderma lucidum spore powder after vacuum drying is ≤ 6%. Then place it at -22 °C for embrittlement for 4.5 h, and then at 50 °C, with a frequency of 30 kHz and a power of 500 W, perform ultrasonic mixing and wall-breaking for 3 min to obtain pretreated Ganoderma lucidum spore powder; S2: Add the pretreated Ganoderma lucidum spore powder to water at a material-liquid ratio of 1:30, mix and stir for 30 min, then perform ultrasonic extraction at 55 °C, a frequency of 40 kHz, and a power of 800 W for 5 min, and then let it stand for 5 h and centrifuge to obtain the first extract; S3: Adjust the pH of the first extract to (8 ± 0.2), add trypsin, perform water bath heating and enzymatic hydrolysis at 35 °C for 1.5 h and then centrifuge to obtain the second extract. The addition amount of trypsin is 0.2% of the mass of the first extract, and the enzyme activity is 200,000 u / g; S4: Add cation exchange resin to the second extract, stir and remove the enzyme for 1.5 h, and filter to obtain the third extract. The addition amount of cation exchange resin is 20 g / L; S5: Concentrate the third extract under vacuum to 1 / 3 of its original volume, then set the vacuum degree to -0.2 MPa for vacuum freeze-drying treatment. First, pre-cool at -16 °C for 12 h, then freeze at -22 °C for 24 h, and finally heat up to 45 °C with a sublimation time of 20 h to obtain the product. Example 3

[0036] A method for preparing Ganoderma lucidum spore polysaccharide, comprising the following steps: S1: Wash and remove impurities from the Ganoderma lucidum spore powder, then perform vacuum drying. Set the vacuum degree to 0.9 MPa and the vacuum drying time to 6 h to make the moisture content of the vacuum-dried Ganoderma lucidum spore powder ≤ 6%. Then place it at -20 °C for embrittlement for 5 h, and then perform ultrasonic mixing and cell wall breaking at 45 - 50 °C, a frequency of 20 kHz, and a power of 300 W for 4 min to obtain the pretreated Ganoderma lucidum spore powder; S2: Add the pretreated Ganoderma lucidum spore powder to water at a material-liquid ratio of 1:25, mix and stir for 25 min, then perform ultrasonic extraction at 50 °C, a frequency of 30 kHz, and a power of 600 W for 6 min, and then let it stand for 6 h and centrifuge to obtain the first extract; S3: Adjust the pH of the first extract to (8 ± 0.2), add trypsin, perform water bath heating and enzymatic hydrolysis at 40 °C for 1.2 h and then centrifuge to obtain the second extract. The addition amount of trypsin is 0.15% of the mass of the first extract, and the enzyme activity is 200,000 u / g; S4: Add cation exchange resin to the second extract, stir and remove the enzyme for 1.2 h, and filter to obtain the third extract. The addition amount of cation exchange resin is 15 g / L; S5: Concentrate the third extract under vacuum to 1 / 3 of its original volume, then set the vacuum degree to -0.2 MPa for vacuum freeze-drying treatment. First, pre-cool at -16 °C for 12 h, then freeze at -22 °C for 24 h, and finally heat up to 45 °C with a sublimation time of 20 h to obtain the product. Comparative example

[0037] Comparative Example 1 A method for preparing Ganoderma lucidum spore polysaccharide, which is different from Example 1, includes the following steps: S1: Wash and remove impurities from Ganoderma lucidum spore powder, then conduct vacuum drying. Set the vacuum degree to 0.9 MPa and the vacuum drying time to 6 h to make the moisture content of the vacuum-dried Ganoderma lucidum spore powder ≤ 6%. Then place it at -19°C for embrittlement for 5.5 h to obtain pretreated Ganoderma lucidum spore powder; S2: Add the pretreated Ganoderma lucidum spore powder to water according to a solid-liquid ratio of 1:20. After mixing and stirring for 20 min, carry out ultrasonic extraction at 45°C at a frequency of 30 kHz and a power of 600 W for 13 min, then let it stand for 6 h and centrifuge to obtain the first extract; S3: Adjust the pH of the first extract to (8 ± 0.2), add trypsin, and carry out water bath heating and enzymatic hydrolysis at 45°C for 1 h and then centrifuge to obtain the second extract. The addition amount of trypsin is 0.1% of the mass of the first extract, and the enzyme activity is 200,000 u / g; S4: Add cation exchange resin to the second extract, stir and remove the enzyme for 1 h, and filter to obtain the third extract. The addition amount of cation exchange resin is 15 g / L; S5: Concentrate the third extract under vacuum to 1 / 3 of the original volume, then set the vacuum degree to -0.2 MPa for vacuum freeze-drying treatment. First pre-cool at -16°C for 12 h, then freeze at -22°C for 24 h, and finally heat up to 45°C and the sublimation time is 20 h to obtain the product.

[0038] Comparative Example 2 A method for preparing Ganoderma lucidum spore polysaccharide, which is different from Example 1, includes the following steps: S1: Wash and remove impurities from Ganoderma lucidum spore powder, then conduct vacuum drying. Set the vacuum degree to 0.9 MPa and the vacuum drying time to 6 h to make the moisture content of the vacuum-dried Ganoderma lucidum spore powder ≤ 6%. Then place it at -19°C for embrittlement for 5.5 h, and then carry out ultrasonic mixing and cell wall breaking at 45°C at a frequency of 20 kHz and a power of 300 W for 5 min to obtain pretreated Ganoderma lucidum spore powder; S2: Add the pretreated Ganoderma lucidum spore powder to water according to a solid-liquid ratio of 1:20. After mixing and stirring for 45 min, let it stand for 6 h and centrifuge to obtain the first extract; S3: Adjust the pH of the first extract to (8 ± 0.2), add trypsin, and carry out water bath heating and enzymatic hydrolysis at 45°C for 1 h and then centrifuge to obtain the second extract. The addition amount of trypsin is 0.1% of the mass of the first extract, and the enzyme activity is 200,000 u / g; S4: Add cation exchange resin to the second extract, stir for 1 hour to remove enzymes, filter to obtain the third extract, and the addition amount of the cation exchange resin is 15 g / L; S5: Concentrate the third extract under vacuum to 1 / 3 of its original volume, then set the vacuum degree to -0.2 MPa for vacuum freeze-drying treatment. First, pre-cool at -16 °C for 12 hours, then freeze at -22 °C for 24 hours, and finally heat up to 45 °C with a sublimation time of 20 hours to obtain the product.

[0039] Comparative Example 3 A method for preparing ganoderma spore polysaccharide, which is different from Example 1 and includes the following steps: S1: Wash and remove impurities from ganoderma spore powder, then perform vacuum drying. Set the vacuum degree to 0.9 MPa and the vacuum drying time to 6 hours to make the moisture content of the vacuum-dried ganoderma spore powder ≤ 6%. Then place it at -19 °C for embrittlement for 5.5 hours, and then at 45 °C, carry out ultrasonic mixing and wall-breaking for 5 minutes at a frequency of 20 kHz and a power of 300 W to obtain pretreated ganoderma spore powder; S2: Add the pretreated ganoderma spore powder to water according to a solid-liquid ratio of 1:20, mix and stir for 20 minutes, then at 45 °C, carry out ultrasonic extraction for 8 minutes at a frequency of 30 kHz and a power of 600 W, then let it stand for 6 hours, and centrifuge to obtain the first extract; S3: Concentrate the first extract under vacuum to 1 / 3 of its original volume, then set the vacuum degree to -0.2 MPa for vacuum freeze-drying treatment. First, pre-cool at -16 °C for 12 hours, then freeze at -22 °C for 24 hours, and finally heat up to 45 °C with a sublimation time of 20 hours to obtain the product. Application Example

[0040] Application of the ganoderma spore polysaccharide prepared by the method for preparing ganoderma spore polysaccharide of the present application in the preparation of eye perimeter milia removal gel.

[0041] Application Example 1 An eye perimeter milia removal gel, the raw materials and their dosages are shown in Table 1, wherein the ganoderma spore polysaccharide is obtained by the preparation method of Example 1, the fruit acid is glycolic acid and citric acid with a mass ratio of 1:0.4, the emulsifier is lecithin and polysorbate-60 with a mass ratio of 1:2.5, and the skin moisturizing component is polydimethylsiloxane and isononyl isononanoate with a mass ratio of 1:1.

[0042] Table 1

[0043] The preparation method of the above-mentioned eye perimeter milia removal gel includes the following steps: S1. Mix the emulsifier with glycerol and squalane after adding water according to the ingredient ratio. After heating to 50 °C, add ganoderma spore polysaccharide, fruit acid, salicylic acid and p - hydroxyacetophenone. Stir evenly and then cool to room temperature to obtain the active stock solution; S2. Add xylitol glucoside, dehydrated xylitol, vitamin B5, shea butter, sorbitan isostearate and skin - moisturizing components to the active stock solution according to the ingredient ratio. After mixing evenly, adjust the pH to (5.5 ± 0.2), stir at 35 °C for 20 min, cool to room temperature and sterilize to obtain the product. Application Example 2

[0044] An eye - area milia removal gel, different from Application Example 1 in that the raw materials and their dosages are shown in Table 1. Among them, the fruit acid is glycolic acid and citric acid with a mass ratio of 1:0.5, the emulsifier is lecithin and polysorbate - 60 with a mass ratio of 1:3, the skin - moisturizing component is polydimethylsiloxane and glyceryl trioctanoate with a mass ratio of 1:1, and other steps are the same as those in Application Example 1. Application Example 3

[0045] An eye - area milia removal gel, different from Application Example 1 in that the raw materials and their dosages are shown in Table 1. Among them, the fruit acid is glycolic acid and citric acid with a mass ratio of 1:0.6, the emulsifier is lecithin and polysorbate - 60 with a mass ratio of 1:4.5, the skin - moisturizing component is isononyl isononanoate, and other steps are the same as those in Application Example 1. Application Example 4

[0046] An eye - area milia removal gel, different from Application Example 1 in that the raw materials and their dosages are shown in Table 1. Among them, the fruit acid is glycolic acid and citric acid with a mass ratio of 1:0.5, the emulsifier is lecithin and polysorbate - 60 with a mass ratio of 1:4, the skin - moisturizing component is isoamyl acetate and glyceryl trioctanoate with a mass ratio of 1:1, and other steps are the same as those in Application Example 1. Application Example 5

[0047] An eye - area milia removal gel, different from Application Example 1 in that the ganoderma spore polysaccharide in the raw materials is obtained by the preparation method of Example 2, and other steps are the same as those in Application Example 1. Application Example 6

[0048] An eye - area milia removal gel, different from Application Example 1 in that the ganoderma spore polysaccharide in the raw materials is obtained by the preparation method of Example 3, and other steps are the same as those in Application Example 1. Comparative Application Example

[0049] Comparative Application Example 1 An eye perimeter milia removal gel, which is different from Application Example 1 in that the ganoderma spore polysaccharide in the raw materials is obtained by the preparation method of Comparative Example 1, and the other steps are the same as those in Application Example 1.

[0050] Comparative Application Example 2 An eye perimeter milia removal gel, which is different from Application Example 1 in that the ganoderma spore polysaccharide in the raw materials is obtained by the preparation method of Comparative Example 2, and the other steps are the same as those in Application Example 1.

[0051] Comparative Application Example 3 An eye perimeter milia removal gel, which is different from Application Example 1 in that the ganoderma spore polysaccharide in the raw materials is obtained by the preparation method of Comparative Example 3, and the other steps are the same as those in Application Example 1.

[0052] Comparative Application Example 4 An eye perimeter milia removal gel, which is different from Application Example 1 in that no ganoderma spore polysaccharide is added, and the ganoderma spore polysaccharide in the raw materials is replaced with glycerol of equal mass, and the other steps are the same as those in Application Example 1. Performance detection test

[0053] Perform the following relevant performance detection tests on the eye perimeter milia removal gels obtained from Application Examples 1 - 6 and Comparative Application Examples 1 - 4.

[0054] Test Example 1 Detection of product physical and chemical properties

[0055] Perform stability tests on the eye perimeter milia removal gels obtained from Application Examples 1 - 6 and Comparative Application Examples 1 - 4 according to the general methods in the cosmetics industry, and record the test results in Table 2.

[0056] Among them, the pH value test is carried out according to the determination of pH value in the "Hygienic Chemical Test Methods" of the "Cosmetics Safety and Technology Specifications" (2015 edition); the heat resistance test is carried out on the product according to the "Heat Resistance" test method in 5.2.2 of GB / T 29665 Skin Care Lotion (o / w type), that is, keep it at (40 ± 1) °C for 24 h, and there is no separation phenomenon after returning to room temperature; the cold resistance test is carried out on the product according to the "Cold Resistance" test method in 5.2.3 of GB / T 29665 Skin Care Lotion (o / w type), that is, keep it at (-8 ± 2) °C for 24 h, and there is no separation phenomenon after returning to room temperature; the centrifugation test is carried out on the product according to the "Centrifugation Test" test method in 5.2.4 of GB / T 29665 Skin Care Lotion (o / w type), that is, 2000 r / min, 30 min, and there is no stratification.

[0057] Table 2

[0058] As can be seen from the performance test results in Table 2, the Ganoderma lucidum spore polysaccharide prepared by the method for preparing Ganoderma lucidum spore polysaccharide of the present application has good stability when applied to the preparation of eye perimeter milia removal gel, and has good stability in heat resistance, cold resistance and centrifugation tests.

[0059] Test Example 2 Detection of Hygiene Index

[0060] Perform hygiene index detections such as microorganisms and heavy metals on the eye perimeter milia removal gel obtained from Application Examples 1-6 and Comparative Application Examples 1-4. The specific item types and technical indicators are shown in Table 3.

[0061] Detect according to the corresponding requirements in the "Cosmetics Safety and Technical Specifications" (2015 Edition). The results show that all the detection indicators of the eye perimeter milia removal gel obtained from Application Examples 1-6 and Comparative Application Examples 1-4 meet the requirements.

[0062] Table 3

[0063] Test Example 3 Detection of Milia Improvement Effect

[0064] Select 55 female volunteers aged between 25 and 55 with milia problems around the eyes and similar quantity and degree as the test subjects. Randomly and evenly divide them into 11 groups, with 5 test subjects in each group. Among them, groups 1-10 are used as experimental groups. After cleaning the skin every day, apply the eye perimeter milia removal gel obtained from Application Examples 1-6 and Comparative Application Examples 1-4 around the eyes of each group respectively. Observe the improvement of the eye perimeter milia after 30 days. The 11th group is used as the blank group and does not use any eye care products.

[0065] Use the VISIA skin image analyzer to take facial images of the testers before and after using the gel. Score the fine lines, milia improvement and eye perimeter skin conditions after using the gel for different items relative to before using (0-100 points). Take the average value of the scoring data of each group as the specific scoring result and record it in Table 4.

[0066] Milia Scoring Criteria: Below 60 points: No obvious improvement, the number of milia does not decrease, and may even increase. 60-80: Slight improvement, the number of milia decreases slightly, the eye perimeter skin is dry, and there is a slight skin rejection reaction of peeling. 80-90: Obvious improvement, the number of milia decreases significantly, the size or color of the milia also improves significantly, the eye perimeter skin is moist, and no skin rejection reaction occurs. 90 - 100: Significant improvement. The number of milia has been greatly reduced, almost all have disappeared or only a very small number remain. The size and color of the remaining milia have also been significantly improved. The eye area skin is moisturized and no skin rejection reaction has occurred.

[0067] Table 4

[0068] From the performance test results of experimental application examples 1 - 6, comparative application example 4 and the blank control group in Table 4, it can be seen that the ganoderma spore polysaccharide prepared by the preparation method of a ganoderma spore polysaccharide of the present application can effectively reduce the possible skin rejection reaction caused by salicylic acid and peracid substances when applied to the preparation of an eye area milia removal gel. It synergizes with active ingredients such as salicylic acid and fruit acid to promote the keratin metabolism of the eye area skin, dissolve sebum accumulation, effectively reduce and eliminate eye area milia, and improve the overall condition of the eye area skin.

[0069] Test Example 4 Detection of the Effect of Improving Dark Circles and Eye Bags

[0070] Select 220 volunteers aged 25 - 50 with relatively deep dark circles and large eye bags as test subjects, with a male - female ratio of 1:1. They are randomly divided into 11 groups, with each group including 10 males and 10 females. Among them, groups 1 - 10 are experimental groups. During the test period, 20 subjects in each group apply an eye area milia removal gel obtained from experimental application examples 1 - 6 and comparative application examples 1 - 4 of the present application twice a day, morning and evening, after cleansing their faces. The 11th group is the blank group and does not apply any eye care products during the test period. The eye area dark circles and eye bag conditions of each group of test subjects are tested and recorded before use and after 8 weeks of use.

[0071] Test Instruments: MDD4 - Mexameter MX 18, Minolta CM - 700D spectrophotometer, Skin Rapid Optical Imaging System PRIMOS lite.

[0072] Test Method: Enter the test environment 30 minutes before the test, clean the skin and wait quietly for 15 minutes for testing; use the CM - 700D spectrophotometer to continuously test the face 6 times and take the average value as the skin color; use the MDD4 - Mexameter MX18 to test the melanin content. This instrument is based on the principle of spectral absorption and determines the content of melanin and heme in the skin by measuring the reflection amount of light with a specific wavelength on the human skin. The smaller the melanin value, the whiter the skin, and vice versa, the darker the skin; use the Skin Rapid Optical Imaging System PRIMOS lite to test the volume of the skin protrusion under the eyes and evaluate the change of the eye bag. The smaller the eye bag volume value, the smaller the eye bag volume.

[0073] The test data measured for all subjects in each group were averaged after removing the maximum and minimum values, and then statistically analyzed. The final results were recorded in Table 5 - 8. SPSS 13.0 software was used for statistical analysis, and repeated - measures analysis of variance, paired t - test, Friedman test, and Wilcoxon test were applied to analyze the data. A P value < 0.05 indicated a statistically significant difference.

[0074] 1. The statistical results of the melanin content before and after using the eye - area milia - removing gel are shown in Table 5, and the skin color test results are shown in Table 6. L * represents brightness (0 - 100), 0 represents black, and 100 represents white. The larger the skin color L * value, the brighter the skin color. The P values of the blank group and the 10 experimental groups were all < 0.05.

[0075] Table 5

[0076] Table 6

[0077] 2. The statistical results of the eye - bag volume before and after using the eye - area milia - removing gel are shown in Table 7 below. The P values of the blank group and the 10 experimental groups were all < 0.05.

[0078] Table 7

[0079] From the performance test results of Application Examples 1 - 6, Comparative Application Example 4, and the blank control group in Tables 5 - 7, it can be seen that the ganoderma spore polysaccharide prepared by the preparation method of a ganoderma spore polysaccharide of the present application can cooperate with effective ingredients such as salicylic acid and fruit acid when applied to prepare the eye - area milia - removing gel, promote the metabolism and blood circulation of the eye - area skin, effectively improve the problems of eye - area dark circles and eye bags, and at the same time brighten the skin color.

[0080] It can be seen from the performance test results of Application Examples 1-6 and Comparative Application Examples 1-2 in Table 4-7 that when the Ganoderma lucidum spore polysaccharide prepared by the preparation methods of Comparative Examples 1-2 is applied to the preparation of an eye perimeter milia removal gel, the improvement effects on eye perimeter dark circles and eye bags and the skin brightening effect are significantly inferior to the application effects of the Ganoderma lucidum spore polysaccharide prepared by the preparation methods of Examples 1-3. This is because for the Ganoderma lucidum spore polysaccharide prepared by the preparation method of Comparative Example 1, ultrasonic treatment was only carried out in the water extraction stage and not in the pretreatment stage, while for the Ganoderma lucidum spore polysaccharide prepared by the preparation method of Comparative Example 2, ultrasonic treatment was only carried out in the pretreatment stage and not in the water extraction stage. Ultrasonic treatment is carried out in different mass transfer mechanisms in the pretreatment stage and water extraction stage of Ganoderma lucidum spore powder to break the hard cell wall of Ganoderma lucidum spore powder, which is more efficient. The ultrasonic treatment under the two mass transfer mechanisms enables the extraction rate of Ganoderma lucidum spore polysaccharide to reach a more balanced level, and at the same time reduces the excessive damage of ultrasonic treatment to the active ingredients in Ganoderma lucidum spore powder, ensuring the preparation efficiency and quality of Ganoderma lucidum spore polysaccharide, and thus ensuring the efficacy of Ganoderma lucidum spore polysaccharide as a raw material for the preparation of an eye perimeter milia removal gel in eliminating and reducing milia, improving eye perimeter dark circles and eye bags, and improving skin color.

[0081] It can be seen from the performance test results of Application Examples 1-6 and Comparative Application Example 3 in Table 4-7 that when the Ganoderma lucidum spore polysaccharide prepared by the preparation method of Comparative Example 3 is applied to the preparation of an eye perimeter milia removal gel, the improvement effects on eye perimeter dark circles and eye bags and the skin brightening effect are significantly inferior to the application effects of the Ganoderma lucidum spore polysaccharide prepared by the preparation methods of Examples 1-3. This is because the Ganoderma lucidum spore polysaccharide prepared by the preparation method of Comparative Example 3 did not use trypsin to treat the extraction solution of Ganoderma lucidum spore powder, and there are a large number of macromolecular protein impurities in the Ganoderma lucidum spore powder extraction solution. These components are difficult to penetrate the skin and play a direct role, and as impurity components, they also have an adverse impact on the efficacy of the gel. Degrading the macromolecular proteins in Ganoderma lucidum spore powder that are difficult to penetrate the skin and play a direct role into small peptides and amino acids. On the one hand, because small peptides and amino acids have a small molecular weight and good skin permeability, they can carry Ganoderma lucidum spore polysaccharide to enter the deep layer of the skin more effectively, thereby exerting better biological activity. On the other hand, small peptides and amino acids can act as auxiliary components to cooperate with Ganoderma lucidum spore polysaccharide, and can significantly improve the effects of the prepared eye perimeter milia removal gel in removing and reducing eye perimeter milia, improving dark circles and eye bags, and brightening the skin.

[0082] This specific embodiment is only an interpretation of the present application and is not a limitation of the present application. Those skilled in the art can make modifications to this embodiment without creative contributions according to needs after reading this specification, but as long as it is within the scope of the claims of the present application, it is protected by the patent law.

Claims

1. A method for preparing Ganoderma lucidum spore polysaccharide, characterized in that: The following steps are involved: S1: washing, removing impurities, and vacuum drying the Ganoderma lucidum spore powder, and then embrittlement at -19 to -22°C for 4.5-5.5h, and then ultrasonically mixing and breaking the spores at 45-50°C for 3-5min to obtain pretreated Ganoderma lucidum spore powder; S2: adding the pretreated Ganoderma lucidum spore powder to water at a solid-liquid ratio of 1: (20-30), mixing and stirring for 20-30 minutes, performing ultrasonic extraction for 5-8 minutes, and then standing for 5-6 hours, centrifuging, and obtaining a first extract; S3: adjusting the pH of the first extract to (8±0.2), adding trypsin, heating in a water bath at 35-45° C. for 1-1.5 h, and then centrifuging to obtain a second extract; S4: adding a cation exchange resin to the second extract, stirring to remove the enzyme for 1-1.5 hours, filtering to obtain a third extract; S5: Concentrate the third extract to 1 / 3 of its original volume under vacuum, and then perform vacuum freeze-drying to obtain the product.

2. The method for preparing Ganoderma lucidum spore polysaccharide according to claim 1, characterized in that The amount of trypsin added is 0.1-0.2% of the mass of the first extract, and the enzyme activity is 200,000 u / g.

3. The method for preparing Ganoderma lucidum spore polysaccharide according to claim 1, characterized in that The cation exchange resin is a macroporous resin with a sulfur group, and the addition amount is 15-20g / L.

4. The method for preparing Ganoderma lucidum spore polysaccharide according to claim 1, characterized in that In step S1, the frequency of ultrasonic mixing and wall breaking is 20-30kHz, the power is 300-500W, and the frequency of ultrasonic extraction in step S2 is 30-40kHz, the power is 600-800W, and the temperature is 45-55℃.

5. A Ganoderma lucidum spore polysaccharide, characterized in that: The polysaccharide is prepared by the method for preparing the polysaccharide from ganoderma lucidum spores according to any one of claims 1 to 4.

6. Use of the Ganoderma lucidum spore polysaccharide as claimed in claim 5 in preparing a gel for removing fat particles around the eyes.

7. A gel for removing fat particles around the eyes, characterized in that: Calculated by mass percentage, the raw materials include 6-8% of the Ganoderma lucidum spore polysaccharide according to claim 5, 1-3% fruit acid, 0.4-0.7% salicylic acid, 0.5-1% p-hydroxyacetophenone, 5-10% glycerin, 2-5% squalane, 2-5% emulsifier, 0.5-1.5% xylitol glucoside, 1-2% anhydroxylitol, 1-2% vitamin B5, 2-5% avocado butter, 0.5-1.5% sorbitan isostearate and 4-6% emollient component, and the balance is water.

8. The gel for removing fat particles around the eyes according to claim 7, characterized in that , the fruit acid is glycolic acid and citric acid in a mass ratio of 1:(0.4-0.6).

9. The gel for removing fat particles around the eyes according to claim 7, characterized in that , the emulsifier is lecithin and polysorbate-60 in a mass ratio of 1:(2.5-4.5).

10. The gel for removing fat particles around the eyes according to claim 7, characterized in that The emollient component is selected from any one or more combinations of polydimethylsiloxane, isononyl isononanoate, isopentyl acetate, and caprylic / capric triglyceride.

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