CN114521652A - Composition with anti-aging effect and application thereof - Google Patents

Abstract

The invention provides a composition with an anti-aging effect and application thereof. The composition with anti-aging effect comprises: schizophyllum commune fermented product, plant extract and collagen peptide. The mechanism of the action of the composition on the aged skin is found to be capable of increasing the contents of hydroxyproline and hyaluronic acid in the skin, improving the activity of antioxidant enzymes of a mouse body and inhibiting the secretion of inflammatory cytokines.

Description

Composition with anti-aging effect and application thereof

Technical Field

The invention belongs to the technical field of health-care food, and particularly relates to a composition with an anti-aging effect and application thereof.

Background

Schizophyllum commune is a precious edible fungus, and the main active substance in the fruiting body, mycelium and fermentation liquor is schizophyllan. Research shows that the compound has pharmacological activities of resisting tumor, bacteria and radiation, improving immunity and the like.

The main application in oral health food is Schizophyllum commune fruiting body extract. However, the wild schizophyllum commune is precious, so that the yield of the artificially cultured sporocarp is limited, and the application range of the wild schizophyllum commune is directly influenced. Meanwhile, the Schizophyllum commune extract is dark in color and has limitations in application to oral products. Although the Schizophyllum commune fermented product (Schizophyllum commune polysaccharide) has application in cosmetics, the application of the Schizophyllum commune fermented product in oral cosmetic products is not seen.

CN113337545A discloses a Schizophyllum commune fermentation product, a preparation method thereof, a skin care product and a Schizophyllum commune culture medium. In particular to a Schizophyllum commune fermentation product, a preparation method thereof, a skin care product and a Schizophyllum commune culture medium. The preparation method of the Schizophyllum commune fermentation product comprises the following steps: culturing Schizophyllum commune in Schizophyllum commune culture medium, and collecting fermentation product; wherein the schizophyllum commune culture medium contains kudzu root. Pueraria powder is added into a culture medium to culture Schizophyllum commune, and a fermentation product is collected, wherein the obtained fermentation product has positive effects on oxidation resistance and free radical removal.

CN107320541A discloses Schizophyllum commune semen ziziphi spinosae syrup and a preparation process thereof, wherein the syrup consists of Schizophyllum commune fermentation liquor, semen ziziphi spinosae extract, a sweetening agent, an acidity regulator, deionized water and a stabilizing agent, and the components are in parts by weight and the addition ratio are as follows: 35-40 parts of schizophyllum commune fermentation liquor, 25-30 parts of spina date seed extracting solution, 5-8 parts of sweetening agent, 0.5-1 part of sour agent, 330 parts of deionized water and 350 parts of stabilizing agent and 1-2 parts of stabilizing agent. The invention adopts the schizophyllum commune fermentation liquor and the wild jujube seed extract to directly prepare the product, avoids the toxic and side effects of the residual organic solvent, is more acceptable to patients when taking the syrup than special medicine taking, can achieve the treatment effect while enjoying the delicious food, and has great economic value and development potential.

CN106173661A discloses a composite strain mixed fermentation functional beverage and a preparation method thereof, wherein each 1000mL of purified water contains: 9-15 g of poria cocos, 10-30 g of dictyophora phalloidea, 6-12 g of tremella aurantialba, 10-20 g of grifola frondosa, 10-30 g of hericium erinaceus, 30-60 g of armillaria mellea, 9-16 g of schizophyllum commune, 1-5 g of sweetening agent, 0.1-0.5 g of acidity regulator, and 0.1-0.5 g of thickening agent and stabilizing agent. The solid content is not less than 10 percent, the total acid content is less than 1.0 percent, the color is yellow brown, uniform and consistent, the precipitate is not easy to occur after long-term storage, the special fragrance and taste of fermentation are provided, the sweet and bitter taste is appropriate, the taste is fine and harmonious, the mouthfeel is full, and the style is unique and pure.

Therefore, the development of a Schizophyllum commune fermented product for use in an oral cosmetic product is a research focus in the field.

Disclosure of Invention

Aiming at the defects of the prior art, the invention aims to provide a composition with an anti-aging effect and application thereof. The mechanism of the composition on the aging skin is found to be capable of increasing the contents of hydroxyproline and hyaluronic acid in the skin, improving the activity of antioxidant enzymes of a mouse body and inhibiting the secretion of inflammatory cytokines.

In order to achieve the purpose, the invention adopts the following technical scheme:

in a first aspect, the present invention provides a composition having anti-aging effect, comprising: schizophyllum commune fermented product, plant extract and collagen peptide.

In the invention, the Schizophyllum commune fermentation product, the plant extract and the collagen peptide are mutually matched, so that the Schizophyllum commune fermentation product has a synergistic effect, has an anti-aging effect on skin, can obviously improve the contents of hydroxyproline and hyaluronic acid in the skin, improves the activity of antioxidant enzymes of a mouse body, and has an inhibition effect on inflammatory cytokine secretion.

Preferably, the composition with the anti-aging effect comprises the following components in parts by weight: 20-40 parts of schizophyllum commune fermentation product, 3-10 parts of plant extract and 7-16 parts of collagen peptide.

The content of the Schizophyllum commune fermented product in the composition is 20-40 parts, such as 20 parts, 22 parts, 24 parts, 26 parts, 28 parts, 30 parts, 32 parts, 34 parts, 36 parts, 38 parts, 40 parts and the like.

In the composition, the plant extract content is 3-10 parts, and may be, for example, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts, etc.

In the composition, the collagen peptide is contained in an amount of 7 to 16 parts, for example, 7 parts, 8 parts, 9 parts, 10 parts, 11 parts, 12 parts, 13 parts, 14 parts, 15 parts, 16 parts, etc.

Preferably, the Schizophyllum commune fermented product is obtained by culturing Schizophyllum commune in a medium containing radix Puerariae.

Wherein the Schizophyllum commune fermented product CN113337545A is prepared by the preparation method disclosed by the patent specification. It has positive effects in resisting oxidation and scavenging free radicals. The provided Schizophyllum commune fermentation product contains Schizophyllum commune polysaccharide as an active ingredient, and shows better capabilities in the aspects of removing excessive free radicals and inhibiting related enzyme activities by combining the antioxidant capacity of the radix puerariae matrix.

The raw material sources and the preparation method of the schizophyllum commune fermentation product are as follows:

firstly, a schizophyllum commune culture medium: 2-8g/L of kudzu root powder (less than or equal to 40 meshes), 0.1-0.5 wt% of yeast extract, 0.01-0.15 wt% of magnesium sulfate, 0.05-0.2 wt% of potassium dihydrogen phosphate, 1-4 wt% of glucose and the balance of water.

In the schizophyllum commune medium, the particle size of the pueraria lobata powder is not more than 40 meshes, and may be, for example, 40 meshes, 35 meshes, 30 meshes, 25 meshes, 20 meshes, 15 meshes, 10 meshes, 5 meshes, 2 meshes, 1 mesh, or the like.

In the schizophyllum commune culture medium, the content of the kudzu root powder is 2-8g/L, for example, 2g/L, 2.5g/L, 3g/L, 3.5g/L, 4g/L, 4.5g/L, 5g/L, 5.5g/L, 6g/L, 6.5g/L, 7g/L, 7.5g/L, 8g/L, etc.

The content of the yeast extract in the Schizophyllum commune culture medium is 0.1-0.5 wt%, and may be, for example, 0.1 wt%, 0.2 wt%, 0.3 wt%, 0.4 wt%, 0.5 wt%, or the like.

The magnesium sulfate content in the Schizophyllum commune culture medium is 0.01-0.15 wt%, and may be, for example, 0.01 wt%, 0.02 wt%, 0.04 wt%, 0.06 wt%, 0.08 wt%, 0.1 wt%, 0.12 wt%, 0.14 wt%, 0.15 wt%, or the like.

In the Schizophyllum commune culture medium, the content of potassium dihydrogen phosphate is 0.05-0.2 wt%, and may be, for example, 0.05 wt%, 0.06 wt%, 0.08 wt%, 0.1 wt%, 0.12 wt%, 0.14 wt%, 0.16 wt%, 0.18 wt%, 0.2 wt%, etc.

The amount of glucose in the Schizophyllum commune medium is 1-4 wt%, and may be, for example, 1 wt%, 2 wt%, 3 wt%, 4 wt%, etc.

② fermentation culture: performing activated culture on Schizophyllum commune strain (GIM 5.43 provided by Guangdong province microbial research institute) to obtain Schizophyllum commune; then 10% of Schizophyllum commune (v/v, Schizophyllum commune seed solution/total volume of the culture medium after seed solution inoculation) is inoculated into the culture medium, and fermentation culture is carried out for 100-150h (for example, 100h, 110h, 120h, 130h, 140h, 150h and the like) under the conditions of 28-30 ℃ (for example, 28 ℃, 29 ℃, 30 ℃ and the like), 140-170r/min (for example, 140r/min, 150r/min, and the like).

Preparing fermentation liquor: and (4) stopping centrifuging the fermentation product, separating the thallus from the fermentation liquor, and obtaining supernatant as the fermentation liquor.

Preferably, the plant extract comprises any one or a combination of at least two of olive extract, roxburgh rose extract, emblic leafflower fruit extract, blueberry extract, cranberry extract or aronia melanocarpa extract.

Preferably, the plant extract is obtained by PEF extraction.

Preferably, the plant extract is prepared by the following preparation method:

(a) extracting the plant raw materials by using water as a solvent through a PEF extraction method to obtain a plant extracting solution;

(b) filtering and concentrating the plant extract obtained in the step (a) to obtain an extract;

(c) and (c) mixing the extract obtained in the step (b) with maltodextrin, and then carrying out spray drying to obtain the plant extract.

Preferably, in step (a), the mass-to-volume ratio of the plant material to water is 1 (10-30) g/mL, for example, the ratio of the mass of the plant material to water may be 1g/10mL, 1g/15mL, 1g/20mL, 1g/25mL, 1g/30mL, or the like.

Preferably, in step (a), the field strength of the extraction is 10-30kV/cm, such as 10kV/cm, 15kV/cm, 20kV/cm, 25kV/cm, 30kV/cm, etc., and the number of pulses of the extraction is 4-12, such as 4, 5, 6, 7, 8, 9, 10, 11, 12, etc.

Preferably, in step (b), 200-300 mesh filter membranes are used for filtration, and can be 200 meshes, 220 meshes, 240 meshes, 260 meshes, 280 meshes, 300 meshes and the like.

Preferably, in step (b), the concentration temperature is 70-90 deg.C, such as 70 deg.C, 75 deg.C, 80 deg.C, 85 deg.C, 90 deg.C.

Preferably, in the step (c), the mass ratio of the extract to the maltodextrin is 1 (0.05-0.4), and may be 1:0.05, 1:0.1, 1:0.15, 1:0.2, 1:0.25, 1:0.3, 1:0.35, 1:0.4, etc.

Preferably, in step (c), the air inlet temperature for spray drying is 85-120 deg.C, such as 85 deg.C, 90 deg.C, 95 deg.C, 100 deg.C, 105 deg.C, 110 deg.C, 115 deg.C, 120 deg.C, etc., and the air outlet temperature is 65-90 deg.C, such as 65 deg.C, 70 deg.C, 75 deg.C, 80 deg.C, 85 deg.C, 90 deg.C, etc.

In a second aspect, the present invention provides the use of a composition having anti-aging effects as described in the first aspect in the preparation of a health food.

Preferably, the health food comprises a solid beverage and/or a liquid beverage.

In a third aspect, the present invention provides a solid beverage comprising the composition having anti-aging effect according to the first aspect.

Preferably, the solid beverage comprises the following components in parts by weight: 20-40 parts of the composition with the anti-aging effect, 0-1 part of a flavoring agent, 0-4.5 parts of edible essence and 15-30 parts of maltodextrin.

In the solid beverage, the content of the composition having an anti-aging effect is 20 to 40 parts, and may be, for example, 20 parts, 22 parts, 24 parts, 26 parts, 28 parts, 30 parts, 32 parts, 34 parts, 36 parts, 38 parts, 40 parts, and the like.

The flavoring agent may be contained in the solid beverage in an amount of 0 to 1 part, and may be, for example, 0 part, 0.1 part, 0.2 part, 0.4 part, 0.6 part, 0.8 part, 1 part, or the like.

In the solid beverage, the content of the flavoring essence is 0-4.5 parts, for example, 0 part, 0.1 part, 0.2 part, 0.4 part, 0.6 part, 0.8 part, 1 part, 1.2 parts, 1.5 parts, 2 parts, 2.5 parts, 3 parts, 3.5 parts, 4 parts, 4.5 parts and the like.

In the solid beverage, the content of the maltodextrin is 15 to 30 parts, and may be, for example, 15 parts, 16 parts, 18 parts, 20 parts, 22 parts, 24 parts, 26 parts, 28 parts, 30 parts, or the like.

In a fourth aspect, the present invention provides a method for preparing a solid beverage as described in the third aspect, specifically comprising the following steps:

(1) mixing the plant extract and maltodextrin, mixing with Schizophyllum commune fermented product, and granulating to obtain granule mixture;

(2) and (2) drying the particle mixture obtained in the step (1), and then totally mixing the particle mixture with collagen peptide, optional flavoring agent and optional flavoring essence to obtain the solid beverage.

Preferably, in step (1), the granulation is performed in a high-efficiency wet granulator, the mixing and stirring speed in the granulation is 500-.

Preferably, in the step (2), the drying temperature is 65-75 deg.C, such as 65 deg.C, 66 deg.C, 68 deg.C, 70 deg.C, 72 deg.C, 74 deg.C, 75 deg.C, etc.

Preferably, in the step (2), the total mixing is carried out in a three-dimensional mixer, and the total mixing is followed by sub-packaging by a multi-column packaging machine.

In a fifth aspect, the present invention provides a liquid beverage comprising the composition having anti-aging effect according to the first aspect.

Preferably, the liquid beverage comprises the following components in parts by weight: 10-30 parts of the composition with the anti-aging effect, 6-15 parts of fruit concentrated juice, 0.01-0.5 part of edible essence, 0.01-1 part of flavoring agent and 20-40 parts of water.

In the liquid beverage, the content of the composition having an anti-aging effect is 10 to 30 parts, and may be, for example, 10 parts, 12 parts, 14 parts, 16 parts, 18 parts, 20 parts, 22 parts, 24 parts, 26 parts, 28 parts, 30 parts, and the like.

In the liquid beverage, the content of the fruit juice concentrate is 6 to 15 parts, and may be, for example, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts, 11 parts, 12 parts, 13 parts, 14 parts, 15 parts, or the like.

In the liquid beverage, the content of the edible essence is 0.01-0.5 parts, for example, 0.01 parts, 0.05 parts, 0.1 parts, 0.2 parts, 0.3 parts, 0.4 parts, 0.5 parts and the like.

The content of the flavoring agent in the liquid beverage is 0.01 to 1 part, and may be, for example, 0.01 part, 0.05 part, 0.1 part, 0.2 part, 0.3 part, 0.4 part, 0.5 part, 0.6 part, 0.8 part, 1 part, or the like.

The content of water in the liquid beverage is 20 to 40 parts, and may be, for example, 20 parts, 22 parts, 24 parts, 26 parts, 28 parts, 30 parts, 32 parts, 34 parts, 36 parts, 38 parts, 40 parts, or the like.

In a sixth aspect, the present invention provides a method for preparing a liquid beverage according to the fifth aspect, specifically comprising the following steps:

(I) mixing the composition with the anti-aging effect, the concentrated fruit juice, the edible essence, the flavoring agent and water, and sieving to obtain a material body;

(II) sterilizing the material body obtained in the step (I), and filling to obtain the liquid beverage.

Preferably, in step (I), the mixing temperature is 65-75 deg.C, such as 65 deg.C, 66 deg.C, 68 deg.C, 70 deg.C, 72 deg.C, 75 deg.C, etc., and time is 10-20min, such as 10min, 12min, 14min, 16min, 18min, 20min, etc.

Preferably, in step (I), the mesh has a pore size of 200-300 meshes, such as 200 meshes, 220 meshes, 240 meshes, 260 meshes, 280 meshes, 300 meshes, etc.

Preferably, in step (II), the sterilization is carried out using ultra-high temperature flash sterilization, UHT, technique;

preferably, in the step (II), the packaging bottle used for filling needs to be cleaned by an anion cleaning technology; and the filling is carried out by adopting a full-automatic filling machine.

The beverage prepared by the product is sterilized by UHT, is different from the conventional heating sterilization, can ensure the main active ingredients of the raw materials, and can ensure the original taste and flavor of the product in shelf life.

Compared with the prior art, the invention has the following beneficial effects:

(1) the invention provides a new application of a Schizophyllum commune fermented product, which is prepared by a fermentation process with a proprietary technology and is applied to an oral beauty product. The composition has remarkable antiaging effect;

(2) the invention researches the mechanism of the Schizophyllum commune fermentation product composition on the action of the skin with aging for the first time, and finds that the Schizophyllum commune fermentation product composition can improve the contents of hydroxyproline and hyaluronic acid in the skin, improve the activity of antioxidant enzyme of a mouse body and inhibit the secretion of inflammatory cytokines. Has obvious synergistic effect.

Detailed Description

The technical solution of the present invention is further explained by the following embodiments. It should be understood by those skilled in the art that the examples are only for the understanding of the present invention and should not be construed as the specific limitations of the present invention.

The following examples and comparative examples have the following sources of the components:

preparation example 1

The preparation example provides a schizophyllum commune fermented product, which is obtained by the following preparation method:

(A) the schizophyllum commune culture medium is prepared according to the following mass percentages: 5g/L of kudzu root powder, 0.3% of yeast extract, 0.05% of magnesium sulfate, 0.1% of monopotassium phosphate, 3% of glucose and the balance of water.

(B) Inoculating 10% Schizophyllum commune (v/v, Schizophyllum commune seed solution/total volume of culture medium after inoculating seed solution) into the above culture medium, and fermenting at 28 deg.C and 150 r/min. After fermenting for 120h, centrifuging the fermentation product, and removing thallus and radix Puerariae powder to obtain the fermentation product.

Preparation example 2

The preparation example provides an olive extract, which is prepared by the following preparation method:

(a) extracting olive raw materials by using water as a solvent through a PEF extraction method to obtain olive extracting solution; wherein the volume ratio of the olive raw material to water is 1g:20mL, the field intensity of extraction is 20kV/cm, and the pulse frequency is 8 times;

(b) filtering and concentrating the olive extracting solution obtained in the step (a) to obtain an extract; wherein, the filtration adopts a 200-mesh filter membrane, and the concentration temperature is 80 ℃;

(c) mixing the extract obtained in the step (b) with maltodextrin, and then carrying out spray drying to obtain the olive extract; wherein the mass ratio of the extract to the maltodextrin is 1:0.2, the air inlet temperature of the spray drying is 100 ℃, and the air outlet temperature is 70 ℃.

Preparation example 3

The preparation example provides a roxburgh rose extract, which is prepared by the following preparation method:

(a) extracting the roxburgh rose raw material by using water as a solvent through a PEF extraction method to obtain roxburgh rose extracting solution; wherein the mass of the roxburgh rose raw material and the volume ratio of water are 1g:10mL, the field intensity of extraction is 15kV/cm, and the pulse frequency is 6 times;

(b) filtering and concentrating the rosa roxburghii tratt extract obtained in the step (a) to obtain an extract; wherein, the filtration adopts a 300-mesh filter membrane, and the concentration temperature is 75 ℃;

(c) mixing the extract obtained in the step (b) with maltodextrin, and then carrying out spray drying to obtain the roxburgh rose extract; wherein the mass ratio of the extract to the maltodextrin is 1:0.1, the air inlet temperature of spray drying is 90 ℃, and the air outlet temperature is 70 ℃.

Preparation example 4

The preparation example provides an emblic extract, which is prepared by the following preparation method:

(a) extracting raw materials of the emblic leafflower fruits by using water as a solvent through a PEF extraction method to obtain an emblic leafflower fruit extracting solution; wherein the volume ratio of the mass of the raw material of the emblic leafflower fruit to the water is 1g:30mL, the field intensity of the extraction is 30kV/cm, and the pulse frequency is 10 times;

(b) filtering and concentrating the emblic extract obtained in the step (a) to obtain an extract; wherein, the filtration adopts a 300-mesh filter membrane, and the concentration temperature is 90 ℃;

(c) mixing the extract obtained in the step (b) with maltodextrin, and then carrying out spray drying to obtain the emblic leafflower fruit extract; wherein the mass ratio of the extract to the maltodextrin is 1:0.3, the air inlet temperature of spray drying is 120 ℃, and the air outlet temperature is 90 ℃.

Preparation example 5

The preparation example provides a blueberry extract, and the blueberry extract is prepared by the following preparation method:

(a) extracting the blueberry raw material by using water as a solvent through a PEF extraction method to obtain a blueberry extracting solution; wherein the volume ratio of the mass of the blueberry raw material to the volume of water is 1g:20mL, the field intensity of extraction is 20kV/cm, and the pulse frequency is 12 times;

(b) filtering and concentrating the blueberry extract obtained in the step (a) to obtain an extract; wherein, the filtration adopts a 200-mesh filter membrane, and the concentration temperature is 80 ℃;

(c) mixing the extract obtained in the step (b) with maltodextrin, and then carrying out spray drying to obtain the blueberry extract; wherein the mass ratio of the extract to the maltodextrin is 1:0.2, the air inlet temperature of the spray drying is 100 ℃, and the air outlet temperature is 70 ℃.

Preparation example 6

The preparation example provides a cranberry extract, and the cranberry extract is prepared by the following preparation method:

(a) extracting a cranberry raw material by using water as a solvent through a PEF extraction method to obtain a cranberry extracting solution; wherein the volume ratio of the mass of the cranberry raw material to the volume of water is 1g:20mL, the field intensity of extraction is 18kV/cm, and the pulse frequency is 10 times;

(b) filtering and concentrating the cranberry extract obtained in the step (a) to obtain an extract; wherein, the filtration adopts a 200-mesh filter membrane, and the concentration temperature is 80 ℃;

(c) mixing the extract obtained in the step (b) with maltodextrin, and then carrying out spray drying to obtain the cranberry extract; wherein the mass ratio of the extract to the maltodextrin is 1:0.2, the air inlet temperature of spray drying is 100 ℃, and the air outlet temperature is 70 ℃.

Preparation example 7

The preparation example provides a aronia melanocarpa extract, which is prepared by the following preparation method:

(a) extracting the aronia melanocarpa raw material by using water as a solvent through a PEF extraction method to obtain an aronia melanocarpa extracting solution; wherein the volume ratio of the mass of the aronia melanocarpa raw material to the water is 1g:15mL, the field intensity of extraction is 25kV/cm, and the pulse frequency is 6 times;

(b) filtering and concentrating the aronia melanocarpa extract obtained in the step (a) to obtain an extract; wherein, the filtration adopts a 300-mesh filter membrane, and the concentration temperature is 80 ℃;

(c) mixing the extract obtained in the step (b) with maltodextrin, and performing spray drying to obtain the aronia melanocarpa extract; wherein the mass ratio of the extract to the maltodextrin is 1:0.1, the air inlet temperature of spray drying is 90 ℃, and the air outlet temperature is 70 ℃.

Example 1

The embodiment provides a composition with an anti-aging effect, which comprises the following components in parts by weight: 30 parts of schizophyllum commune fermentation product provided in preparation example 1, 5 parts of olive extract provided in preparation example 2, and 10 parts of collagen peptide.

Example 2

The embodiment provides a composition with an anti-aging effect, which comprises the following components in parts by weight: 25 parts of schizophyllum commune fermentation product provided in preparation example 1, 8 parts of rosa roxburghii tratt extract provided in preparation example 3 and 12 parts of collagen peptide.

Example 3

The embodiment provides a composition with an anti-aging effect, which comprises the following components in parts by weight: 30 parts of schizophyllum commune fermentation product provided in preparation example 1, 5 parts of blueberry extract provided in preparation example 4 and 10 parts of collagen peptide.

Example 4

The embodiment provides a composition with an anti-aging effect, which comprises the following components in parts by weight: 32 parts of schizophyllum commune fermentation product provided by preparation example 1, 3 parts of blueberry extract provided by preparation example 5 and 7 parts of collagen peptide.

Example 5

The embodiment provides a composition with an anti-aging effect, which comprises the following components in parts by weight: 40 parts of schizophyllum commune fermentation product provided in preparation example 1, 3 parts of cranberry extract provided in preparation example 6, and 7 parts of collagen peptide.

Example 6

The embodiment provides a composition with an anti-aging effect, which comprises the following components in parts by weight: 20 parts of schizophyllum commune fermented product provided in preparation example 1, 10 parts of aronia melanocarpa extract provided in preparation example 7, and 16 parts of collagen peptide.

Example 7

This example provides a composition with anti-aging effect, which is different from example 1 only in that the schizophyllum commune fermented substance provided in preparation example 1 is replaced by the schizophyllum commune fermented liquid provided in example 1 in CN 107320541A.

Example 8

This example provides a composition having anti-aging effect, which is different from example 1 only in that the olive extract provided in preparation example 2 is replaced with a commercially available olive extract (manufacturer: Nippon Biopsis, cat # CB-2021091).

Comparative example 1

This comparative example provides a composition comprising, in parts by weight: 33 parts of Schizophyllum commune fermentation product and 12 parts of collagen peptide provided in preparation example 1.

Comparative example 2

This comparative example provides a composition comprising, in parts by weight: 35 parts of the Schizophyllum commune fermented product provided in preparation example 1 and 10 parts of the olive extract provided in preparation example 2.

Comparative example 3

This comparative example provides a composition comprising, in parts by weight: 20 parts of olive extract and 25 parts of collagen peptide provided in preparation example 2.

Test example 1

Ability to scavenge free radicals

Test samples: examples 1-8 provide compositions having anti-aging effects and comparative examples 1-3;

the test method comprises the following steps:

1. evaluation of superoxide anion radical scavenging ability

4.5mL of Tris-HCl buffer (0.05 mol/LpH ═ 8.2) was preheated in a water bath at 25 ℃ for 20 min. Then adding 1mL of sample and 0.4mL of 25mmol/L pyrogallol solution, mixing uniformly, reacting in a water bath at 25 ℃ for 5min, and adding 1.0mL of 8mol/L HCl to terminate the reaction. Absorbance values were measured at 299nm with Tris-HCl buffer as a reference. Blank a 1mL aliquot of solvent was used in place of the sample.

Superoxide anion radical scavenging rate (%) ([ 1- (A)₂/A₁)]X is 100%; in the formula A₁Absorbance values for the blank; a. the₂Is the absorbance value of the sample.

2. Evaluation of hydroxyl radical scavenging ability

Sequentially adding 2mmol/L FeSO into a 25mL colorimetric tube₄ 3mL，1mmol/L H₂O₂3mL, shaking up, then adding 3mL of salicylic acid with the concentration of 6mmol/L, shaking up, heating in a water bath at 37 ℃ for 15min, taking out, and measuring the absorbance; adding the solutions to be tested with certain concentrations, shaking, heating in water bath for 15min, and taking out to test their absorbance. The following formula is the clearance rate of hydroxyl radical (. OH) by the liquid to be tested:

hydroxyl radical clearance (%) ═ a₁-A₂-(A₁-A₃)]/A₁X is 100%; in the formula A₁The absorbance value of the reaction system before adding the medicine is obtained; a. the₂Removing OH from the sample to obtain the absorbance value of the system; a. the₃Removing OH for blank control to obtain absorbance value of the system;

3. evaluation of scavenging ability for DPPH free radical

Test A20 mmol/L DPPH solution was prepared according to the method specified in Larrauri JA; test sample solution: the schizophyllan compositions provided in examples 1-10 and the compositions provided in comparative examples 1-5 were diluted with ultrapure water to give solutions having a mass concentration of 0.1%. Putting 2.0mL of test sample solution and 2.0mL of 20mmol/L DPPH solution in a test tube, mixing uniformly, reacting for 30min, measuring absorbance value at 517nm, taking absolute ethyl alcohol as blank control, and calculating DPPH inhibition ratio according to the absorbance value.

DPPH radical inhibition (%) - [1- (A)₁-A₂)/A₃]X is 100%; wherein A is₁Absorbance of 2.0mL of DPPH solution and 2.0mL of test sample solution, A₂Absorbance of 2.0mL of the test sample solution and 2.0mL of absolute ethanol, A₃The absorbance of 2.0mL of DPPH solution and 2.0mL of absolute ethyl alcohol is obtained, and the average value is obtained by parallel testing for three times;

the specific test results are shown in table 1:

TABLE 1

As shown in the test data in Table 1, the composition with the anti-aging effect has good oxidation resistance and DPPH free radical removing capacity, the superoxide anion free radical removing rate is more than 60%, the hydroxyl free radical removing rate is more than 60%, and the DPPH free radical inhibiting rate is more than 80%.

As can be seen from the comparison between example 1 and comparative examples 1-3, the three components of the composition with anti-aging effect cooperate with each other to have a synergistic effect, so that the scavenging effect of the composition on DPPH free radicals, hydroxyl free radicals and superoxide anion free radicals can be further improved, the anti-oxidation capability of skin tissues can be enhanced, and the free radical content of the skin tissues can be reduced.

Test example 2

Detection of hydroxyproline

Test samples: examples 1-8 provide compositions having anti-aging effects and comparative examples 1-3;

the test method comprises the following steps:

1. preparing human skin cells

Taking dermal fibroblast of normal human as primary cell at 75cm²In a culture flask (2 mL of growth factor-containing medium), CO at 37 ℃₂Cells were cultured at a concentration of 5% to a concentration of 80%. Passage is carried out, and the passage cell is 175cm²The cells were continued to culture to 80% concentration in the flask.

2. Preparation of samples to be tested

Culturing human dermal fibroblasts obtained by the previous step in a DMEM culture solution containing 10% fetal calf serum at 37 ℃ for 24 hours in 5% CO2, digesting the cells with a digestive juice containing 0.25% trypsin and 0.02% EDTA after the cells are nearly fused, and then carrying out passage 1:2 to select fourth generation cells;

cells were diluted to density of 10 with DMEM medium containing 10% fetal bovine serum⁵One cell/mL, and then inoculated into 12-well plates at an inoculum size of 2X 10⁵Culturing each cell/well until the cells are nearly 80% fused, washing the plate with physiological saline for 3 times, adding the sample to be tested (0.1 mL of the above sample + 0.2mL of physiological saline) and the control sample (0.3mL of physiological saline) into each well, and culturing at 37 deg.C with 5% CO₂Under the condition of culturingAfter day, culture supernatants of the test sample and the control sample were collected for use.

3. Determination of hydroxyproline content

According to a human hydroxyproline enzyme-linked immunoassay kit (Shanghai enzyme-linked biosciences, Ltd.), hydroxyproline content determination of culture supernatants of a sample to be tested and a control group sample is carried out:

(1) drawing a hydroxyproline standard curve, and preparing 6 hydroxyproline standard solutions, wherein the concentrations are as follows in sequence: 0 mu g/mL, 3 mu g/mL, 6 mu g/mL, 12 mu g/mL, 24 mu g/mL, 48 mu g/mL, and testing the OD value of the sample at the wavelength of 450nm by using an enzyme labeling instrument;

(2) according to the result of the absorbance test of the hydroxyproline standard, drawing a hydroxyproline standard curve by taking the absorbance as a vertical axis and the concentration of hydroxyproline as a horizontal axis;

(3) diluting the culture supernatants of the sample to be detected and the control group sample by 5 times by using a solvent in the kit, determining the absorbance of the culture supernatants of the sample to be detected and the control group sample diluted by 5 times at the wavelength of 450nm according to the specification of a human hydroxyproline enzyme-linked immunoassay kit, and calculating the content of hydroxyproline generated by fibroblasts in the culture supernatants of the sample to be detected and the control group sample according to a drawn hydroxyproline concentration standard curve;

the specific test results are shown in table 2:

TABLE 2

As can be seen from the test data in Table 2, the content of hydroxyproline is obviously increased after the composition with the anti-aging effect is added, the content of hydroxyproline is more than 34 mu g/mL, and the increase rate is more than 25% compared with that of a control group. The composition with the anti-aging effect can obviously improve the amount of collagen secreted by fibroblasts of the skin.

Test example 3

Detection of hyaluronic acid

Test samples: examples 1-8 provide compositions having anti-aging effects and comparative examples 1-3 provide compositions;

the test method comprises the following steps:

measurement of hydroxyproline content in the culture supernatants of each test sample and control sample in test example 2 according to a human hyaluronic acid enzyme-linked immunoassay kit (shanghai enzyme-linked biotechnology limited):

1. drawing a hyaluronic acid standard curve; preparing 6 hyaluronic acid standard substance solutions, wherein the concentrations are as follows in sequence: 0. mu.g/mL, 10. mu.g/mL, 20. mu.g/mL, 40. mu.g/mL, 80. mu.g/mL, 160. mu.g/mL, and the OD value thereof was measured at 450nm using a microplate reader,

2. according to the test result of the hyaluronic acid standard product, drawing a standard curve of hyaluronic acid by taking the absorbance as a vertical axis and taking the concentration of hyaluronic acid as a horizontal axis;

(3) diluting the culture supernatants of the sample to be detected and the control group sample by 5 times by using a solvent in the kit, and calculating the content of hyaluronic acid according to the OD value of the culture supernatants of the human hyaluronic acid enzyme-linked immunoassay kit;

the specific test results are shown in table 3:

TABLE 3

As shown in the test data in Table 2, the content of hyaluronic acid is obviously increased after the composition with the anti-aging effect is added, the content of hyaluronic acid is more than 185 mu g/mL, and the increase rate is more than 40% compared with a control group. The composition with the anti-aging effect can obviously improve the amount of hyaluronic acid secreted by skin fibroblasts, improve the water locking capacity of the skin and keep the skin moist.

Test example 4

Influence on the Activity of antioxidant enzymes in mouse serum

Test samples: examples 1-8 provide compositions having anti-aging effects and comparative examples 1-3;

the test method comprises the following steps:

1. laboratory animal

12-month old SPF-grade healthy mice, female and male half.

2. Experimental methods

Randomly dividing 12-month-old SPF-grade healthy mice into 12 groups, wherein each group comprises 10 mice; wherein 11 groups respectively fed 2.0g/kg per day of the corresponding group of examples 1 to 8 and comparative examples 1 to 3, respectively, in addition to the normal feeding of the conventional feeds, provided the compositions having anti-aging effects; wherein, 1 group is used as control group and is fed with conventional feed normally; after feeding each group continuously for 30 days, the mice were sacrificed and then serum Malondialdehyde (MDA), reduced glutathione content (GSH), and antioxidant enzyme activity were measured in vivo.

3. Statistical treatment

Statistical processing was performed using SPSS11.0 statistical software, data were measured to represent one-way analysis of variance using multiple sample mean comparisons, and count data were tested using x 2.

4. Results of the experiment

Three positive indexes of the four indexes of lipid oxidation products, protein oxidation products, antioxidase and antioxidant substances can determine that the animal experiment result of the antioxidant function of the tested sample is positive.

The specific test results are shown in table 4:

TABLE 4

As can be seen from the test data in Table 4, the MDA level in the serum of the mice taking the composition with the anti-aging effect is remarkably reduced, and the GSH content, the SOD activity and the GSH-Px activity of the antioxidant substances are remarkably increased, wherein the MDA content is less than 4.5mmol/L, the GSH content is more than 5.6mmol/L, the SOD content is more than 280 mu/mL, and the GSH-Px content is more than 200 mu/mL. The composition with the anti-aging effect has good anti-oxidation and health-care effects.

Test example 5

Inhibition of inflammatory cytokine secretion

Test samples: examples 1-8 provide compositions having anti-aging effects and comparative examples 1-3;

the test principle is as follows: "proinflammatory cytokine (proinflammatory cytokine)" refers to a series of cytokines that promote inflammation, and plays an important role in acute inflammatory response, increasing vascular permeability, and causing red, swelling, heat, pain and other inflammatory responses. The more common proinflammatory cytokines include tumor necrosis factor alpha (TNF-alpha), interleukin 6(interleukin-6, IL-6) and interleukin 12(interleukin-12, IL-12), etc., so that the inhibition of these cytokines can also contribute to the reduction of the inflammatory symptoms of certain diseases;

the test method comprises the following steps:

1. test materials: LPS was purchased from Sigma; FBS was purchased from PAA, USA; DMEM high glucose medium; ELISA kits were purchased from biolignd, usa.

2. Animals: c57BL/6 mice, purchased from Beijing Huafukang Biotech GmbH.

3. Experimentally RAW264.7 cells were plated at 3X 10⁵cells/well concentration seeded in 96-well microtiter plates at 37 ℃ 5% CO₂Culturing in an incubator. After 24 hours of incubation, 1. mu.g/mL LPS and sample were added, respectively, followed by 5% CO at 37 ℃₂The culture was carried out in an incubator for 24 hours, in which samples were supplied to the composition having antiaging action according to examples 1 to 8 and the composition according to comparative examples 1 to 3, respectively (sample concentration of 50. mu.g/mL).

After the culture, the culture solution is sucked out, the expression quantity of the proinflammatory cytokines such as TNF-alpha, IL-6, IL-12 and the like is analyzed by using an ELISA kit, and the inhibition rate of the proinflammatory cytokine production of the sample is calculated according to the following formula:

inhibition rate (%) of proinflammatory cytokine production [1-C₁/C₀]×100％；

Wherein, C₀Represents the concentration of proinflammatory cytokine production with addition of the inducer LPS alone, C₁Represents the concentration of proinflammatory cytokines produced when the inducer LPS is added simultaneously with the sample;

the specific test results are shown in table 5:

TABLE 5

As can be seen from the data tested in Table 5, LPS stimulated the activation of BMDM cells, which was shown to secrete a number of inflammatory cytokines TNF- α, IL-6, IL-12. The composition with the anti-aging effect can obviously inhibit the secretion of TNF-alpha, IL-6 and IL-12, and has an inhibition rate of more than 20 percent for TNF-alpha, more than 50 percent for IL-6 and more than 30 percent for IL-12.

Application example 1

The application example provides a solid beverage, which comprises the following components in parts by weight:

name (R)	Proportioning parts
		Example 1 provides compositions with anti-aging action	30
Flavoring agent	0.5
		Edible essence	1
Maltodextrin	20

The preparation method of the solid beverage comprises the following steps:

(1) placing the olive extract and maltodextrin in a high-efficiency wet granulator to carry out wet mixing for 6min, and controlling the mixing and stirring speed to be 150 r/min; then adding Schizophyllum commune fermented product, mixing for 5min, granulating, and controlling the rotation speed of granulating slurry to 800 r/min;

(2) drying the prepared granular material by a fluidized bed to obtain a granular mixture, and controlling the drying temperature to be 70 ℃; then putting the particle mixture, the collagen powder, the sweetener and the edible essence into a three-dimensional mixer for total mixing to obtain a mixture; and finally, subpackaging the total mixed powder by using a multi-row subpackaging machine to control the net content of the product.

Application examples 2 to 8

The present application example provides different solid beverages, differing from application example 1 only in that the composition having an antiaging effect provided in example 1 was replaced with the corresponding composition having an antiaging effect provided in examples 2 to 8.

Comparative application examples 1 to 3

Comparative application example different solid beverages were provided, differing from application example 1 only in that the composition with anti-ageing effect provided in example 1 was replaced by the corresponding compositions provided in comparative examples 1 to 3.

Application example 9

The application example provides a liquid beverage, which comprises the following components in parts by weight:

name (R)	Proportioning parts
		Example 1 provides compositions with anti-aging action	20
Fruit concentrated juice	10
		Edible essence	0.2
Flavoring agent	0.1
		Purified water	30

The preparation method of the liquid beverage comprises the following steps:

(I) preparation: putting the composition with the anti-aging effect, the fruit concentrated juice, the edible essence and the flavoring agent, which are provided by the embodiment 1, into a preparation tank, adding purified water with the formula amount, controlling the temperature at 70 ℃, stirring for 15min until the mixture is uniformly mixed, and then sieving the mixture into a storage tank;

(II) sterilizing the prepared material body by adopting an ultrahigh temperature instantaneous sterilization UHT technology; the inner packaging bottle is cleaned by adopting an anion cleaning technology; and (4) subpackaging the liquid by adopting a full-automatic filling machine.

Application examples 10 to 16

This application example provides different liquid beverages, differing from application example 9 only in that the composition with anti-aging effect provided in example 1 was replaced with the corresponding composition with anti-aging effect provided in examples 2 to 8.

Comparative application examples 4 to 6

Comparative application example different liquid beverages were provided, differing from application example 1 only in that the composition with anti-ageing effect provided in example 1 was replaced by the corresponding compositions provided in comparative examples 1 to 3.

Test example 6

Test for anti-aging Properties

Test samples: examples 1-8 provide compositions having anti-aging effects and comparative examples 1-3 provide compositions;

test methods (zebrafish SA- β -gal staining experiment):

after the zebra fish in the test group or the blank group is dyed, an image is captured and collected by a microscope, all the images uniformly adopt the same shooting parameters including body position, magnification factor, light intensity, absorbance, exposure setting (each sample is respectively exposed for 5ms, 15ms and 30ms, three pictures are shot) and the like, and the acquired image of the zebra fish SA-beta-gal dyeing effect shows that the SA-beta-gal dyeing positive rate of the zebra fish is visually reduced compared with that of the blank control group after the test group is respectively treated by different samples through microscopic imaging.

To ensure comparability, then adopting Image analysis soft Image Pro Plus to respectively obtain a lateral fish body area value (S) and a staining integral optical density value (IOD), and calculating the ratio IOD/S of the lateral fish body area value (S) and the staining integral optical density value (IOD);

the specific test results are shown in table 6:

TABLE 6

As shown in the test data in Table 6, the SA-beta-gal staining positive rate is low in the test group compared with the blank group, and the difference is statistically significant (P < 0.05). The composition with the anti-aging effect can reduce the dyeing positive rate of SA-beta-gal staining of the zebra fish under a proper proportion concentration, and can effectively interfere the aging of embryonic developmental cells of the zebra fish and delay the aging process of the zebra fish, thereby proving that the composition with the anti-aging effect has the anti-aging effect.

The applicant states that the composition with anti-aging effect and the application thereof of the present invention are illustrated by the above examples, but the present invention is not limited to the above examples, that is, the present invention is not meant to be necessarily dependent on the above examples for implementation. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.

Claims (10)

1. A composition with anti-aging effect, which is characterized by comprising the following components: schizophyllum commune fermented product, plant extract and collagen peptide.

2. The composition with anti-aging effect according to claim 1, characterized by comprising the following components in parts by weight: 20-40 parts of schizophyllum commune fermentation product, 3-10 parts of plant extract and 7-16 parts of collagen peptide.

3. The composition with anti-aging effect according to claim 1 or 2, wherein the Schizophyllum commune fermented product is a Schizophyllum commune fermented product obtained by culturing Schizophyllum commune in a medium containing radix Puerariae.

4. The composition with an anti-aging effect according to any one of claims 1 to 3, wherein the plant extract comprises any one of or a combination of at least two of olive extract, rosa roxburghii extract, emblica officinalis extract, blueberry extract, cranberry extract and aronia melanocarpa extract.

5. The composition with anti-aging effect according to any one of claims 1 to 4, wherein the plant extract is obtained by PEF extraction;

preferably, the plant extract is prepared by the following preparation method:

(a) extracting the plant raw materials by using water as a solvent through a PEF extraction method to obtain a plant extracting solution;

(b) filtering and concentrating the plant extract obtained in the step (a) to obtain an extract;

(c) mixing the extract obtained in the step (b) with maltodextrin, and performing spray drying to obtain the plant extract;

preferably, in the step (a), the mass volume ratio of the plant raw material to the water is 1 (10-30) g/mL;

preferably, in the step (a), the field intensity of the extraction is 10-30kV/cm, and the pulse number of the extraction is 4-12 times;

preferably, in the step (b), a 200-mesh and 300-mesh filter membrane is adopted for filtration;

preferably, in step (b), the temperature of the concentration is 70-90 ℃;

preferably, in the step (c), the mass ratio of the extract to the maltodextrin is 1 (0.05-0.4);

preferably, in the step (c), the inlet air temperature of the spray drying is 85-120 ℃, and the outlet air temperature is 65-90 ℃.

6. Use of the composition having an antiaging effect as claimed in any one of claims 1 to 5 for producing a health food;

preferably, the health food comprises a solid beverage and/or a liquid beverage.

7. A solid beverage comprising the composition having an antiaging effect according to any one of claims 1 to 5;

preferably, the solid beverage comprises the following components in parts by weight: 20-40 parts of the composition with the anti-aging effect, 0-1 part of a flavoring agent, 0-4.5 parts of edible essence and 15-30 parts of maltodextrin.

8. A method for preparing a solid beverage according to claim 7, comprising the following steps:

(1) mixing the plant extract and maltodextrin, mixing with Schizophyllum commune fermented product, and granulating to obtain granule mixture;

(2) drying the particle mixture obtained in the step (1), and then totally mixing the particle mixture with collagen peptide, optional flavoring agent and optional edible essence to obtain the solid beverage;

preferably, in the step (1), the granulation is performed in a high-efficiency wet granulator, the mixing and stirring speed in the granulation is 500-1000r/min, and the mixing time is 3-5 min;

preferably, in the step (2), the drying temperature is 65-75 ℃;

preferably, in the step (2), the total mixing is performed in a three-dimensional mixer, and the total mixing further comprises sub-packaging by a multi-row racking machine.

9. A liquid beverage comprising the composition having an anti-aging effect according to any one of claims 1 to 5;

preferably, the liquid beverage comprises, in parts by weight: 10-30 parts of the composition with the anti-aging effect, 6-15 parts of fruit concentrated juice, 0.01-0.5 part of edible essence, 0.01-1 part of flavoring agent and 20-40 parts of water.

10. A method for preparing a liquid beverage according to claim 9, characterized in that it comprises in particular the steps of:

(I) mixing the composition with the anti-aging effect, the concentrated fruit juice, the edible essence, the flavoring agent and water, and sieving to obtain a material body;

(II) sterilizing the material body obtained in the step (I), and filling to obtain the liquid beverage;

preferably, in the step (I), the temperature of the mixing is 65-75 ℃ and the time is 10-20 min;

preferably, in step (I), the aperture of the screen is 200-300 meshes;

preferably, in step (II), the sterilization is carried out using ultra-high temperature flash sterilization, UHT, technique;

preferably, in the step (II), the packaging bottle used for filling needs to be cleaned by an anion cleaning technology; and the filling is carried out by adopting a full-automatic filling machine.