Composition containing schizophyllan as well as preparation method and application thereof

Abstract

The invention relates to a composition containing schizophyllan and a preparation method and application thereof, the composition comprises schizophyllan and a plant extract, and the plant extract is prepared from sambucus chinensis, citrus aurantium, gynura divaricata, waxberry and perigord truffle. The composition prepared by the invention is powder and does not contain preservatives, the addition amount is easy to control, and the composition is easy to store and transport, and effective components in the plant composition can improve the moisturizing and repairing effects of the composition while improving the stability of the composition. Sambucus chinensis, finger citron, Gynura divaricata, waxberry and perigord truffle have a certain synergistic effect on the effects.

Description

Technical Field

[0001] The invention belongs to the technical field of cosmetics and relates to a composition containing schizophyllan and a preparation method and application thereof. Background Art

[0002] Schizophyllan is a bioactive polysaccharide extracted from the fungus Schizophyllan. It has many excellent physiological activities and functional properties, and has shown extremely high application value in many fields such as medicine, skin care, and health care. The advantages of Schizophyllan aqueous solution are that it is easy to use, easy to apply, and quickly absorbed by the skin, which allows Schizophyllan to contact the skin relatively quickly and begin to work. However, the aqueous dosage form also has significant limitations. From the perspective of concentration, due to the physical properties of the aqueous solution and the requirements for formula stability, the concentration of Schizophyllan added to the aqueous solution is difficult to reach a high level, which also limits the intensity of the effect it can produce. In addition, the stability of the aqueous solution is worse than that of the powder. In order to ensure its solubility and stability, it is often necessary to add some specific chemical reagents such as solubilizers. These additional ingredients may interfere with the activity of Schizophyllan itself to a certain extent and affect its efficacy.

[0003] The limitations of existing aqueous dosage forms in practical applications make it difficult to meet the growing demand for efficacy. Powdered dosage forms, with their unique advantages, have shown great potential in the effective utilization and efficacy of Schizophyllan, and are expected to become a better dosage form choice. Therefore, there is an urgent need to provide a Schizophyllan powder composition. Summary of the invention

[0004] In view of the deficiencies of the prior art, the object of the present invention is to provide a composition containing Schizophyllan and a preparation method and application thereof.

[0005] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0006] In a first aspect, the present invention provides a composition containing Schizophyllan, wherein the composition comprises Schizophyllan and a plant extract, wherein raw materials for preparing the plant extract comprise Sambucus sabdariffa, Finger lime, Moonflower, Myrica rubra, and Black truffle.

[0007] Preferably, the composition comprises 1-3 parts of Schizophyllan and 4-8 parts of plant extract by mass.

[0008] The mass proportion of schizophyllan can be selected as 1 part, 1.2 parts, 1.4 parts, 1.6 parts, 1.8 parts, 2 parts, 2.2 parts, 2.4 parts, 2.6 parts, 2.8 parts, 3 parts etc., and the mass proportion of plant extract can be selected as 4 parts, 4.2 parts, 4.5 parts, 4.8 parts, 5 parts, 5.2 parts, 5.5 parts, 5.8 parts, 6 parts, 6.2 parts, 6.5 parts, 6.8 parts, 7 parts, 7.2 parts, 7.5 parts, 7.8 parts, 8 parts etc. Other specific points within the above numerical range can be selected, so they will not be elaborated here one by one.

[0009] Preferably, the raw materials for preparing the plant extract include, by weight, 1-2 parts of sagebrush, 0.5-1 parts of finger lime, 1-2 parts of moonflower, 0.5-1 parts of bayberry, and 0.5-1 parts of black truffle.

[0010] The mass proportion of comfrey can be selected as 1 part, 1.1 parts, 1.2 parts, 1.3 parts, 1.4 parts, 1.5 parts, 1.6 parts, 1.7 parts, 1.8 parts, 1.9 parts, 2 parts, etc., the mass proportion of finger lime can be selected as 0.5 parts, 0.6 parts, 0.7 parts, 0.8 parts, 0.9 parts, 1 parts, etc., the mass proportion of moonflower can be selected as 1 part, 1.1 parts, 1.2 parts, 1.3 parts, 1.4 parts, 1.5 parts, 1.6 parts, 1.7 parts, 1.8 parts, 1.9 parts, 2 parts, etc., the mass proportion of bayberry can be selected as 0.5 parts, 0.6 parts, 0.7 parts, 0.8 parts, 0.9 parts, 1 parts, etc., the mass proportion of black truffle can be selected as 0.5 parts, 0.6 parts, 0.7 parts, 0.8 parts, 0.9 parts, 1 parts, etc., and other specific point values ​​within the above numerical range can be selected, so they will not be elaborated here one by one.

[0011] Preferably, the plant extract is prepared by a method comprising the following steps: mixing the preparation raw material with water, extracting, centrifuging, and filtering.

[0012] Preferably, the mass ratio of the preparation raw materials to water is 3.5:(10-12).

[0013] The specific point values ​​in (10-12) can be selected as 10.5, 11, 11.5, 12, etc. Other specific point values ​​within the above numerical range can also be selected, and they will not be described one by one here.

[0014] Preferably, the leaching temperature is 75-85°C and the leaching time is 0.5-1.5h.

[0015] The temperature can be selected as 75℃, 76℃, 77℃, 78℃, 79℃, 80℃, 81℃, 82℃, 83℃, 84℃, 85℃, etc. The time can be selected as 0.5h, 0.6h, 0.7h, 0.8h, 0.9h, 1h, 1.1h, 1.2h, 1.3h, 1.4h, 1.5h, etc. Other specific point values ​​within the above numerical range can be selected, so they will not be elaborated here.

[0016] Preferably, the pore size of the ceramic membrane used for filtration is 100-150nm, for example 100nm, 105nm, 110nm, 115nm, 120nm, 125nm, 130nm, 135nm, 140nm, 145nm, 150nm, etc. Other specific point values ​​within the above numerical range can be selected, which will not be described here one by one.

[0017] Preferably, the Schizophyllan is prepared by a method comprising the following steps:

[0018] (1) activating the Schizophyllum fungus and then fermenting it to obtain a fermentation liquid;

[0019] (2) concentrating the fermentation liquid to obtain a concentrated liquid;

[0020] (3) Mix the concentrated solution with ethanol, centrifuge to obtain the precipitate, dissolve the precipitate and filter to obtain the product.

[0021] Preferably, the fermentation is divided into an initial fermentation stage, a middle fermentation stage and a late fermentation stage.

[0022] Preferably, the initial fermentation stage refers to within 36 hours from the start of fermentation (for example, 2h, 5h, 10h, 15h, 20h, 25h, 30h, 36h, etc.), the mid-fermentation stage refers to 36-84h of fermentation (for example, 40h, 45h, 50h, 55h, 60h, 65h, 70h, 75h, 80h, 84h, etc.), and the late fermentation stage refers to 84-114h of fermentation (for example, 85h, 90h, 95h, 100h, 105h, 110h, 114h, etc.).

[0023] Other specific point values ​​within the above numerical range can be selected and will not be described in detail here.

[0024] Preferably, the ventilation volume at the initial stage of fermentation is 0.2-0.8 vvm, the dissolved oxygen is 35-45%, and the rotation speed is 100-400 rpm.

[0025] The ventilation volume can be selected as 0.2vvm, 0.3vvm, 0.4vvm, 0.5vvm, 0.6vvm, 0.7vvm, 0.8vvm, etc., the dissolved oxygen can be selected as 35%, 38%, 40%, 42%, 45%, etc., the rotation speed can be selected as 100rpm, 150rpm, 200rpm, 250rpm, 300rpm, 350rpm, 400rpm, etc., and other specific point values ​​within the above numerical range can be selected, so they will not be elaborated here.

[0026] Preferably, the fermentation temperature in the initial stage of fermentation is 26-28°C, for example, 26°C, 26.5°C, 27°C, 27.5°C, 28°C, etc. Other specific points within the above numerical range can be selected and will not be described here one by one.

[0027] Preferably, the ventilation volume in the mid-fermentation stage is 0.5-1.5 vvm, the dissolved oxygen is 30-35%, and the rotation speed is 200-350 rpm.

[0028] The ventilation volume can be selected as 0.5vvm, 0.8vvm, 1vvm, 1.2vvm, 1.5vvm, etc., the dissolved oxygen can be selected as 30%, 31%, 32%, 33%, 34%, 35%, etc., the rotation speed can be selected as 200rpm, 220rpm, 250rpm, 280rpm, 300rpm, 320rpm, 350rpm, etc. Other specific point values ​​within the above numerical range can be selected, so they will not be elaborated here.

[0029] Preferably, the fermentation temperature in the middle fermentation stage is 26-28°C and the pH is 3.8-4.5.

[0030] The temperature can be selected as 26°C, 26.5°C, 27°C, 27.5°C, 28°C, etc., and the pH can be selected as 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, etc. Other specific point values ​​within the above numerical range can be selected, so they will not be described here one by one.

[0031] Preferably, the mid-fermentation stage also includes 3-5 feeding times, for example, 3 times, 4 times, 5 times, etc.

[0032] Preferably, the feeding specifically includes: when the concentration of glucose is reduced to 15-18 g / L (for example, 15 g / L, 15.5 g / L, 16 g / L, 16.5 g / L, 17 g / L, 17.5 g / L, 18 g / L, etc.), adding glucose to a concentration of 20-28 g / L (20 g / L, 21 g / L, 22 g / L, 23 g / L, 24 g / L, 25 g / L, 26 g / L, 27 g / L, 28 g / L, etc.), other specific point values ​​within the above numerical range can be selected, and they will not be described one by one here.

[0033] Preferably, the fermentation medium comprises, by mass percentage, 1-3% glucose, 0.5-2% tryptone, 0.1-1% yeast extract powder, 0.01-0.1% magnesium sulfate, 0.05-0.2% potassium dihydrogen phosphate and water.

[0034] The mass percentage of glucose can be 1%, 1.5%, 2%, 2.5%, 3%, etc. The mass percentage of tryptone can be 0.5%, 0.8%, 1%, 1.2%, 1.5%, 1.8%, 2%, etc. The mass percentage of yeast extract can be 0.1%, 0.2%, 0.5%, 0.8%, 1%, etc. The mass percentage of magnesium sulfate can be 0.01%, 0.02%, 0.05%, 0.08%, 0.1%, etc. The mass percentage of potassium dihydrogen phosphate can be 0.05%, 0.08%, 0.1%, 0.12%, 0.15%, 0.18%, 0.2%, etc. Other specific point values ​​within the above numerical range can be selected, which will not be repeated here.

[0035] Preferably, the fermentation temperature in the late fermentation stage is 35-45°C and the pH is 4.5-5.5.

[0036] The temperature can be selected as 35℃, 36℃, 37℃, 38℃, 39℃, 40℃, 41℃, 42℃, 43℃, 44℃, 45℃, etc., and the pH can be selected as 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, etc. Other specific point values ​​within the above numerical range can be selected, so they will not be elaborated here.

[0037] Preferably, the rotation speed in the late fermentation stage is 250-400 rpm, the ventilation volume is 0.5-1 vvm, and the dissolved oxygen is 25-30%.

[0038] The rotation speed can be selected as 250rpm, 280rpm, 300rpm, 320rpm, 350rpm, 380rpm, 400rpm, etc., the ventilation volume can be selected as 0.5vvm, 0.6vvm, 0.7vvm, 0.8vvm, 0.9vvm, 1vvm, etc., and the dissolved oxygen can be selected as 25%, 26%, 27%, 28%, 29%, 30%, etc. Other specific point values ​​within the above numerical range can be selected, so they will not be elaborated here.

[0039] Preferably, the molecular weight of the ultrafiltration membrane used for filtration is 50-100 kDa, for example 50 kDa, 60 kDa, 70 kDa, 80 kDa, 90 kDa, 100 kDa, etc. Other specific points within the above numerical range can be selected and will not be described one by one here.

[0040] Preferably, the volume of the concentrate is 25-50% of the volume of the fermentation liquid, for example 25%, 30%, 35%, 40%, 45%, 50%, etc. Other specific values ​​within the above numerical range can be selected and will not be described here one by one.

[0041] Preferably, the final concentration of ethanol is 50-70%, for example 50%, 52%, 55%, 58%, 60%, 62%, 65%, 68%, 70%, etc. Other specific points within the above numerical range can be selected and will not be described here one by one.

[0042] Preferably, the dosage form of the composition comprises a powder.

[0043] In a second aspect, the present invention provides a method for preparing the composition containing Schizophyllan according to the first aspect, the preparation method comprising: mixing the plant extract with Schizophyllan, and drying.

[0044] Preferably, the drying temperature is 100-120° C. and the drying time is 60-80 h.

[0045] The temperature can be selected as 100℃, 102℃, 105℃, 108℃, 110℃, 112℃, 115℃, 118℃, 120℃, etc., for example, 60h, 62h, 65h, 68h, 70h, 72h, 75h, 78h, 80h, etc. Other specific point values ​​within the above numerical range can be selected, so they will not be described one by one here.

[0046] In a third aspect, the present invention provides a use of the composition according to the first aspect in preparing a product with moisturizing and repairing effects.

[0047] Compared with the prior art, the present invention has the following beneficial effects:

[0048] The composition prepared by the present invention is a powder, does not contain preservatives, the amount of addition is easy to control, and it is easy to store and transport. The effective ingredients in the plant composition improve the stability of the composition and can also improve the moisturizing and repairing effect of the composition. Elderberry, finger lime, moonflower, bayberry, and black truffle have a certain synergistic effect on the above effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1 It is the test result of the relative abundance of AQP3 gene. DETAILED DESCRIPTION

[0050] In order to further explain the technical means and effects adopted by the present invention, the technical solution of the present invention is further described below in conjunction with the preferred embodiments of the present invention, but the present invention is not limited to the scope of the embodiments.

[0051] The fermentation medium includes, by mass percentage, 2% glucose, 1% tryptone, 0.5% yeast extract powder, 0.05% magnesium sulfate, 0.1% potassium dihydrogen phosphate and the balance water, and is sterilized at 121° C. for 20 minutes.

[0052] The Schizophyllum involved in the following content is the GIM 5.43 strain provided by Guangdong Institute of Microbiology.

[0053] Preparation Example 1

[0054] This preparation example provides a plant extract, which includes, by weight, 1.5 parts of sagebrush, 0.8 parts of finger lime, 1.5 parts of moonflower, 0.8 parts of bayberry, 0.8 parts of black truffle and 17 parts of water.

[0055] The preparation method thereof comprises:

[0056] Mix elderberry, finger lime, moonflower, bayberry, black truffle and water, extract at 80℃ for 1h, and pass the supernatant through a 120nm ceramic membrane to obtain the product.

[0057] Preparation Example 2

[0058] This preparation example provides a plant extract, which includes, by weight, 1 part of sagebrush, 1 part of finger lime, 1 part of moonflower, 1 part of bayberry, 1 part of black truffle and 17 parts of water.

[0059] The preparation method thereof comprises:

[0060] Mix elderberry, finger lime, moonflower, bayberry, black truffle and water, extract at 75℃ for 1.5h, and pass the supernatant through a 100nm ceramic membrane to obtain the product.

[0061] Preparation Example 3

[0062] This preparation example provides a plant extract, which includes, by weight, 2 parts of sagebrush, 0.5 parts of finger lime, 2 parts of moonflower, 0.5 parts of bayberry, 0.5 parts of black truffle and 18 parts of water.

[0063] The preparation method thereof comprises:

[0064] Mix elderberry, finger lime, moonflower, bayberry, black truffle and water, extract at 85℃ for 0.5h, take the supernatant and pass it through a 150nm ceramic membrane to obtain the product.

[0065] Preparation Example 4

[0066] This preparation example provides a Schizophyllan polysaccharide, which is prepared by a method comprising the following steps:

[0067] (1) activating the Schizophyllum sp. by using a PDA medium and then culturing it to obtain a seed solution;

[0068] (2) 10% of the seed liquid was added to the fermentation tank, wherein 0-36h was the initial fermentation stage, and the fermentation conditions were controlled as follows: stirring speed of 200rpm, ventilation volume of 0.5vvm, temperature of 27°C, and dissolved oxygen controlled at 40%;

[0069] 37-84h is the mid-fermentation stage, and the fermentation conditions are controlled as follows: stirring speed 300rpm, adjusting the pH of the fermentation liquid to 4, ventilation volume 1vvm, dissolved oxygen controlled at 33%, temperature 27°C, when glucose drops to 15-18g / L, glucose is added to the fermenter until the glucose content in the fermenter reaches more than 20g / L;

[0070] 85-96h is the late fermentation stage, and the fermentation conditions are controlled as follows: stirring speed 350rpm, pH of the fermentation liquid adjusted to 5, temperature 40°C, ventilation volume 0.8vvm, dissolved oxygen controlled at 28%; after the end of the late stage, fermentation is continued for another 15h;

[0071] (3) After the fermentation is completed, the fermentation liquid is concentrated to 1 / 3 of the original volume, and ethanol is added for precipitation. The final concentration of ethanol is 60%. The precipitate is redissolved with distilled water, and then filtered through an ultrafiltration membrane with a molecular weight of 80 kDa to remove proteins and small molecules, and the schizophyllan is concentrated.

[0072] Preparation Example 5

[0073] This preparation example provides a Schizophyllan polysaccharide, which is prepared by a method comprising the following steps:

[0074] (1) activating the Schizophyllum sp. by using a PDA medium and then culturing it to obtain a seed solution;

[0075] (2) adding 10% of the seed liquid into the fermentation tank, wherein 0-36h is the initial fermentation stage, and the fermentation conditions are controlled as follows: stirring speed of 100rpm, ventilation volume of 0.2vvm, temperature of 26°C, and dissolved oxygen controlled at 35%;

[0076] 37-84h is the mid-fermentation stage, and the fermentation conditions are controlled as follows: stirring speed 200rpm, pH of the fermentation liquid adjusted to 3.8, ventilation volume 0.5vvm, dissolved oxygen controlled at 30%, temperature 26°C, when glucose drops to 15-18g / L, glucose is added to the fermenter until the glucose content in the fermenter reaches more than 20g / L;

[0077] 85-96h is the late fermentation stage, and the fermentation conditions are controlled as follows: stirring speed 250rpm, pH of the fermentation liquid adjusted to 4.5, temperature 35°C, ventilation volume 0.5vvm, dissolved oxygen controlled at 25%; after the end of the late stage, fermentation is continued for another 12h;

[0078] (3) After the fermentation is completed, the fermentation liquid is concentrated to 1 / 4 of the original volume, and ethanol is added for precipitation. The final concentration of ethanol is 50%. The precipitate is redissolved in distilled water, and then filtered through an ultrafiltration membrane with a molecular weight of 50 kDa to remove proteins and small molecules, and the schizophyllan is concentrated.

[0079] Preparation Example 6

[0080] This preparation example provides a Schizophyllan polysaccharide, which is prepared by a method comprising the following steps:

[0081] (1) activating the Schizophyllum sp. by using a PDA medium and then culturing it to obtain a seed solution;

[0082] (2) 10% of the seed liquid was added to the fermentation tank, wherein 0-36h was the initial fermentation stage, and the fermentation conditions were controlled as follows: stirring speed of 400rpm, ventilation volume of 0.8vvm, temperature of 28°C, and dissolved oxygen controlled at 45%;

[0083] 37-84h is the mid-fermentation stage, and the fermentation conditions are controlled as follows: stirring speed 350rpm, pH of the fermentation liquid adjusted to 4.5, ventilation volume 1.5vvm, dissolved oxygen controlled at 35%, temperature 28°C, when glucose drops to 15-18g / L, glucose is added to the fermenter until the glucose content in the fermenter reaches more than 20g / L;

[0084] 85-96h is the late fermentation stage, and the fermentation conditions are controlled as follows: stirring speed 400rpm, pH of the fermentation liquid adjusted to 5.5, temperature 45°C, ventilation volume 1vvm, dissolved oxygen controlled at 30%; after the end of the late stage, continue fermentation for another 18h;

[0085] (3) After the fermentation is completed, the fermentation liquid is concentrated to 1 / 2 of the original volume, and ethanol is added for precipitation. The final concentration of ethanol is 70%. The precipitate is redissolved with distilled water, and then filtered through an ultrafiltration membrane with a molecular weight of 100 kDa to remove proteins and small molecules, and the schizophyllan is concentrated.

[0086] Preparation Example 7

[0087] This preparation example provides a plant extract, which differs from Preparation Example 1 only in that the preparation method is different.

[0088] The preparation method is as follows: mix elderberry, finger lime, moonflower, bayberry, black truffle and water, extract at 80°C for 1 hour, take the supernatant and pass it through a 120nm ceramic membrane, and dry it to obtain the product.

[0089] Preparation Example 8

[0090] This preparation example provides a schizophyllan, which differs from preparation example 4 only in the preparation method.

[0091] The step (3) of the preparation method is "after the fermentation is completed, the fermentation liquid is concentrated to 1 / 3 of the original volume, ethanol is added for precipitation, the final concentration of ethanol is 60%, the precipitate is redissolved with distilled water, and then an ultrafiltration membrane with a molecular weight of 80 kDa is used to filter out proteins and small molecules, and the schizophyllan is concentrated and dried to obtain", and the other operations remain unchanged.

[0092] Comparative Preparation Example 1

[0093] This comparative preparation example provides a plant extract, which differs from Preparation Example 1 only in that it does not contain elderberry, and its reduced mass is proportionally distributed to the mass of finger lime, moonflower, bayberry, and black truffle, while other ingredients and contents remain unchanged.

[0094] The preparation method is similar to that of Preparation Example 1.

[0095] Comparative Preparation Example 2

[0096] This comparative preparation example provides a plant extract, which differs from Preparation Example 1 only in that finger lime is not contained, and its reduced mass is proportionally distributed to the mass of elderberry, moonflower, bayberry, and black truffle, while other components and contents remain unchanged.

[0097] The preparation method is similar to that of Preparation Example 1.

[0098] Comparative Preparation Example 3

[0099] This comparative preparation example provides a plant extract, which differs from Preparation Example 1 only in that it does not contain moonflower, and its reduced mass is proportionally distributed to the mass of elderberry, finger lime, bayberry, and black truffle, while other ingredients and contents remain unchanged.

[0100] The preparation method is similar to that of Preparation Example 1.

[0101] Comparative Preparation Example 4

[0102] This comparative preparation example provides a plant extract, which differs from Preparation Example 1 only in that it does not contain bayberry, and its reduced mass is proportionally distributed to the mass of elderberry, finger lime, moonflower, and black truffle, while other ingredients and contents remain unchanged.

[0103] The preparation method is similar to that of Preparation Example 1.

[0104] Comparative Preparation Example 5

[0105] This comparative preparation example provides a plant extract, which differs from Preparation Example 1 only in that it does not contain black truffle, and its reduced mass is proportionally distributed to the mass of elderberry, finger lime, moonflower, and bayberry, while other ingredients and contents remain unchanged.

[0106] The preparation method is similar to that of Preparation Example 1.

[0107] Example 1

[0108] This example provides a composition containing Schizophyllan, which includes 2 parts of Schizophyllan prepared in Preparation Example 4 and 6 parts of the plant extract prepared in Preparation Example 1, and the parts by mass are all parts by mass of the solutes in the solution.

[0109] The preparation method comprises: mixing schizophyllan with plant extracts, and drying at 110°C for 70 hours to obtain the product.

[0110] Example 2

[0111] This example provides a composition containing Schizophyllan, which includes 1 part of Schizophyllan prepared in Preparation Example 5 and 4 parts of the plant extract prepared in Preparation Example 2 in terms of mass fractions, and the mass fractions are all mass fractions of the solutes in the solution.

[0112] The preparation method comprises: mixing schizophyllan with plant extracts, and drying at 100 DEG C for 80 hours to obtain the product.

[0113] Example 3

[0114] This example provides a composition containing Schizophyllan, which includes 1 part of Schizophyllan prepared in Preparation Example 6 and 8 parts of the plant extract prepared in Preparation Example 3, and the mass fractions are all mass fractions of the solutes in the solution.

[0115] The preparation method comprises: mixing schizophyllan with plant extracts, and drying at 120°C for 60 hours to obtain the product.

[0116] Example 4

[0117] This embodiment provides a composition containing Schizophyllan, which is different from Embodiment 1 only in the preparation method.

[0118] The preparation method thereof is as follows: Schizophyllan polysaccharide is mixed with plant extract to obtain the product.

[0119] Comparative Examples 1-5

[0120] This comparative example provides 5 compositions containing Schizophyllan, which differ from Example 1 only in that the plant extract prepared in Preparation Example 1 is replaced by the plant extracts prepared in Comparative Preparation Examples 1-5 in equal amounts, and other components and contents remain unchanged.

[0121] The preparation method is similar to that of Example 1.

[0122] Test Example 1

[0123] Stability test

[0124] The obtained products were placed at a temperature of 25±2°C and a relative humidity of 60±5% for different periods of time, and the appearance of the products was observed with the naked eye. The results are shown in Table 1.

[0125] Table 1

[0126]

[0127]

[0128] It can be seen from the data in Table 1 that the stability of the powder dosage form is better than that of the liquid dosage form, and the addition of plant extracts improves the stability of the product. Elderflower, finger lime, moonflower, bayberry, and black truffle have a certain synergistic effect in improving product stability.

[0129] Test Example 2

[0130] Cytotoxicity:

[0131] Samples to be tested: samples prepared in Examples 1-4, Comparative Examples 1-5, and Preparation Examples 1, 4, 7, and 8, which were placed at 25° C. and a relative humidity of 65% for 3 months; the liquid samples after being placed for 3 months were freeze-dried to obtain solid powders.

[0132] BJ cells were placed in DMEM high-glucose medium containing 10% fetal bovine serum and 1% double antibody, and cultured at 37°C and 5% carbon dioxide. When the cells were growing well, 1×10 5 The cells were inoculated in a 96-well plate at a density of 100 μL of culture medium per well and incubated at 37°C for 24 h. Five parallel replicates were set for each sample. Except for the blank group, the culture medium containing 1% of the sample to be tested was replaced and cultured for 24 h. Then, 15 μL of CCK-8 reagent was added to each well and incubated at 37°C in the dark for 0.5 h. The OD value was detected at 450 nm to calculate the cell viability.

[0133] Among them: the OD of the normal group is the absorbance of the cells when no sample to be tested is added to the culture medium, and the OD of the blank group is the absorbance of the 96-well plate itself.

[0134]

[0135] Table 2

[0136] Survival rate (%) Example 1 98.01 Example 2 95.41 Example 3 95.26 Example 4 94.53 Comparative Example 1 93.47 Comparative Example 2 92.86 Comparative Example 3 93.14 Comparative Example 4 92.17 Comparative Example 5 92.85 Preparation Example 1 91.76 Preparation Example 4 92.14 Preparation Example 7 93.26 Preparation Example 8 93.14 Blank group 100.00

[0137] It can be seen from the data in Table 2 that the composition of the present application has low cytotoxicity and high safety.

[0138] Test Example 3

[0139] Proliferation effect detection

[0140] Samples to be tested: samples prepared in Examples 1-4, Comparative Examples 1-5, and Preparation Examples 1, 4, 7, and 8, which were placed at 25° C. and a relative humidity of 65% for 3 months; the liquid samples after being placed for 3 months were freeze-dried to obtain solid powders.

[0141] BJ cells were placed in DMEM high-glucose medium containing 10% fetal bovine serum and 1% double antibody, and cultured at 37°C and 5% carbon dioxide. When the cells were growing well, 1×10 4 The density of cells was 5 replicates per concentration. BJ cells were inoculated in a 96-well plate, 150 μL of culture medium was added to each well, and incubated at 37°C for 24 h. Except for the blank group, the culture medium containing 1% of the sample to be tested was replaced and cultured for 24 h. Then, 15 μL of CCK-8 reagent was added to each well, incubated at 37°C in the dark for 0.5 h, and the OD value was detected at 450 nm to calculate the proliferation rate.

[0142]

[0143] Table 3

[0144] Proliferation rate (%) Example 1 118.25 Example 2 115.36 Example 3 114.98 Example 4 107.37 Comparative Example 1 108.83 Comparative Example 2 110.32 Comparative Example 3 109.26 Comparative Example 4 110.54 Comparative Example 5 110.18 Preparation Example 1 103.46 Preparation Example 4 105.26 Preparation Example 7 108.47 Preparation Example 8 109.58 Blank group 100.00

[0145] Test Example 4

[0146] Cell Repair

[0147] Samples to be tested: samples prepared in Examples 1-4, Comparative Examples 1-5, and Preparation Examples 1, 4, 7, and 8, which were placed at 25° C. and a relative humidity of 65% for 3 months; the liquid samples after being placed for 3 months were freeze-dried to obtain solid powders.

[0148] BJ cells were placed in DMEM high-glucose medium containing 10% fetal bovine serum and 1% double antibody, and cultured at 37°C and 5% carbon dioxide. 5 Each well was inoculated with 2 mL of culture medium and incubated at 37°C for 24 h. After making a cross scratch, the scratch distance was measured. Except for the blank group, the culture medium containing 1% of the sample to be tested was replaced, and the scratch distance was measured again after culturing for 24 h.

[0149]

[0150] Table 4

[0151] Repair rate (%) Example 1 19.76 Example 2 17.25 Example 3 16.94 Example 4 12.78 Comparative Example 1 13.38 Comparative Example 2 14.61 Comparative Example 3 14.14 Comparative Example 4 14.35 Comparative Example 5 13.96 Preparation Example 1 9.25 Preparation Example 4 10.36 Preparation Example 7 12.89 Preparation Example 8 14.21 Blank group 6.35 10% culture medium 8.16

[0152] It can be seen from the experimental results in Table 4 that the present application has a good cell repair effect. After being placed for a period of time, the efficacy of the composition in the form of a powder is better than that of the aqueous solution, indicating that the powder is more stable than the aqueous solution. The addition of the plant composition can not only improve the stability, but also improve the repair efficacy of the composition, and there is a certain synergistic effect between the several components.

[0153] Test Example 5

[0154] Moisturizing evaluation

[0155] Samples to be tested: samples prepared in Examples 1-4, Comparative Examples 1-5, and Preparation Examples 1, 4, 7, and 8, which were placed at 25° C. and a relative humidity of 65% for 3 months; the liquid samples after being placed for 3 months were freeze-dried to obtain solid powders.

[0156] BJ cells were placed in DMEM high-glucose medium containing 10% fetal bovine serum and 1% double antibody, and cultured at 37°C and 5% carbon dioxide. 5 The density of cells was 2 mL per well in a 6-well plate and incubated at 37°C for 24 hours. The culture medium was replaced with a culture medium containing 1% of the sample to be tested and treated for 24 hours. RNA in the cells was extracted using TRIzol reagent, and the mRNA was reverse transcribed into cDNA after the RNA concentration was determined. The gene level of AQP3 synthesis expression of the test molecule was detected by fluorescent quantitative PCR as a template. Each sample was tested three times independently, and the results were presented as mean ± standard deviation.

[0157] The reference correction gene was β-actin (ACTB); the blank group consisted of cells treated with medium containing 2% FBS for 24 hours.

[0158] Depend on Figure 1 The results show that the composition prepared in the present application has a good moisturizing effect. After being placed for a period of time, the efficacy of the composition in the powder form is better than that of the aqueous solution, indicating that the powder is more stable than the aqueous solution. The addition of the plant composition can not only improve the stability, but also improve the moisturizing effect of the composition, and there is a certain synergistic effect between the several ingredients.

[0159] The applicant declares that the present invention illustrates a composition containing Schizophyllan and its preparation method and application through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0160] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0161] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A composition containing Schizophyllan, characterized in that The composition comprises schizophyllan and plant extracts, and the raw materials for preparing the plant extracts comprise schizonepeta tenuifolia, finger lime, moonflower, bayberry and black truffle.

2. The composition containing Schizophyllan according to claim 1, characterized in that The composition comprises 1-3 parts of Schizophyllan and 4-8 parts of plant extract by weight; Preferably, the raw materials for preparing the plant extract include, by weight, 1-2 parts of sagebrush, 0.5-1 parts of finger lime, 1-2 parts of moonflower, 0.5-1 parts of bayberry, and 0.5-1 parts of black truffle.

3. The composition containing Schizophyllan according to claim 1 or 2, characterized in that The plant extract is prepared by a method comprising the following steps: mixing the raw material with water for extraction, centrifuging, and filtering to obtain the plant extract.

4. The composition containing Schizophyllan according to claim 3, characterized in that The mass ratio of the preparation raw materials to water is 3.5:(10-12); Preferably, the leaching temperature is 75-85°C and the leaching time is 0.5-1.5h; Preferably, the pore size of the ceramic membrane used for filtration is 100-150 nm.

5. The composition containing Schizophyllan according to any one of claims 1 to 4, characterized in that The schizophyllan is prepared by a method comprising the following steps: (1) activating the Schizophyllum fungus and then fermenting it to obtain a fermentation liquid; (2) concentrating the fermentation liquid to obtain a concentrated liquid; (3) Mix the concentrated solution with ethanol, centrifuge to obtain the precipitate, dissolve the precipitate and filter to obtain the product.

6. The composition containing Schizophyllan according to claim 5, characterized in that The fermentation is divided into an initial fermentation stage, a mid-fermentation stage and a late fermentation stage; Preferably, the initial fermentation stage refers to within 36 hours from the start of fermentation, the middle fermentation stage refers to 36-84 hours of fermentation, and the late fermentation stage refers to 85-114 hours of fermentation; Preferably, the ventilation volume at the initial stage of fermentation is 0.2-0.8 vvm, the dissolved oxygen is 35-45%, and the rotation speed is 100-400 rpm; Preferably, the fermentation temperature in the initial stage of fermentation is 26-28°C; Preferably, the ventilation volume in the mid-fermentation stage is 0.5-1.5 vvm, the dissolved oxygen is 30-35%, and the rotation speed is 200-350 rpm; Preferably, the fermentation temperature in the mid-fermentation stage is 26-28°C and the pH is 3.8-4.5; Preferably, the fermentation temperature in the late fermentation stage is 35-45°C and the pH is 4.5-5.5; Preferably, the rotation speed in the late fermentation stage is 250-400 rpm, the ventilation volume is 0.5-1 vvm, and the dissolved oxygen is 25-30%; Preferably, the molecular weight of the ultrafiltration membrane used for filtration is 50-100 kDa; Preferably, the volume of the concentrate is 25-50% of the volume of the fermentation broth; Preferably, the final concentration of ethanol is 50-70%.

7. The composition containing Schizophyllan according to any one of claims 1 to 6, characterized in that The dosage form of the composition includes powder.

8. A method for preparing a composition containing Schizophyllan according to any one of claims 1 to 7, characterized in that: The preparation method comprises: mixing the plant extract with Schizophyllan and drying the mixture to obtain the product.

9. The method for preparing a composition containing Schizophyllan according to claim 8, characterized in that: The drying temperature is 100-120° C. and the drying time is 60-80 hours.

10. Use of the composition containing Schizophyllan according to any one of claims 1 to 7 in the preparation of a product with moisturizing and repairing effects.

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