CN113520942B

CN113520942B - Anti-oxidation and anti-aging five-ginseng fermentation product and preparation method and application thereof

Info

Publication number: CN113520942B
Application number: CN202110814318.3A
Authority: CN
Inventors: 何敬愉; 陈慧芳
Original assignee: Guangzhou Huanya Cosmetic Science and Technology Co Ltd
Current assignee: Guangzhou Huanya Cosmetic Science and Technology Co Ltd
Priority date: 2021-07-19
Filing date: 2021-07-19
Publication date: 2022-12-06
Anticipated expiration: 2041-07-19
Also published as: CN113520942A

Abstract

The invention belongs to the technical field of cosmetics, and discloses an antioxidant and anti-aging five-ginseng fermented product, and a preparation method and application thereof. The preparation method of the five-ginseng fermented product comprises the following steps: 1) Homogenizing the five-ginseng raw material under high pressure to obtain a five-ginseng stock solution; the five ginseng materials include ginseng, salvia miltiorrhiza, radix sophorae flavescentis, adenophora stricta, radix scrophulariae and walnut kernels; 2) Carrying out enzymolysis on the five-ginseng stock solution by using hydrolase to obtain a five-ginseng hydrolysate; 3) Fermenting the five-ginseng hydrolysate by using the fermentation strain to obtain the five-ginseng fermented product. The invention effectively increases the types and the contents of active ingredients in the five-ginseng fermented product by organically combining high-pressure homogenization, enzymolysis reaction and microbial fermentation technologies, and particularly improves the contents of the active ingredients, wherein the ginsenoside Rb1 is 18.9-28.8 mu g/mL, the salvianolic acid B is 311-386 mu g/mL, the matrine is 51-64 mu g/mL, and the oxymatrine is 52-76 mu g/mL.

Description

Anti-oxidation and anti-aging five-ginseng fermentation product and preparation method and application thereof

Technical Field

The invention belongs to the technical field of cosmetics, and particularly relates to an anti-oxidation and anti-aging five-ginseng fermentation product as well as a preparation method and application thereof.

Background

The prescription of Puji emphasizes the treatment based on syndrome differentiation, and treats dermatosis based on the differentiation of qi and blood of zang-fu organs. According to the record of the general prescription, the Wushen Yuan adopts ginseng, salvia miltiorrhiza, radix sophorae flavescentis, adenophora stricta and figwort root to mix, 15 g of walnut is mixed with various medicines to be smashed into pills, and tea water is used for treating wine thorn and face sore. In ancient applications, the formulation was administered primarily as a powder. Based on the skin care effect of the raw materials in the prescription, active ingredients are mostly extracted by a water decoction method at present and used as health care products or cosmetics. The prior art discloses a Wushen Yuanyuan fermented protoplasm cosmetic and a preparation method and application thereof. The preparation method comprises the following steps: (1) preparation of Wu Canyuan dry powder: mixing Ginseng radix, saviae Miltiorrhizae radix, radix Sophorae Flavescentis, radix Adenophorae, radix scrophulariae, semen Juglandis, and green tea, drying, pulverizing, and sieving to obtain Wu Canyuan dry powder; (2) preparation of Wu Can round fermentation initial system: mixing the yeast liquid with the Wu Canyuan dry powder obtained in the step (1) and water to obtain a Wu Can circular fermentation initial system; (3) preparation of Wu Can round fermentation primary stock solution: performing fermentation culture on the Wu Can round fermentation initial system obtained in the step (2) to obtain Wu Can round fermentation initial stock solution; (4) preparation of Wu Can round fermentation raw pulp: sterilizing the Wu Can round fermentation primary stock solution obtained in the step (3), then centrifuging, and taking the centrifuged supernatant to obtain Wu Can round fermentation raw stock. The Wu Can round fermentation raw pulp obtained by the method has low content of active ingredients.

Disclosure of Invention

The present invention has been made to solve at least one of the above-mentioned problems occurring in the prior art. Therefore, the invention provides a preparation method of an anti-oxidation and anti-aging five-ginseng fermented product, which can obtain the five-ginseng fermented product with high content of active ingredients.

The invention provides a preparation method of an antioxidant and anti-aging five-ginseng fermented product, which comprises the following steps:

1) Homogenizing the five-ginseng raw material under high pressure to obtain a five-ginseng stock solution; the five ginseng raw materials comprise ginseng, salvia miltiorrhiza, radix sophorae flavescentis, adenophora stricta, radix scrophulariae and walnut kernels; the pressure of the high-pressure homogenization is 40-100MPa;

2) Carrying out enzymolysis on the five-ginseng stock solution by using hydrolase to obtain five-ginseng hydrolysate;

3) And fermenting the five-ginseng hydrolysate by using a fermentation strain to obtain the five-ginseng fermented product.

According to modern medical research, the main components of ginseng are ginsenosides, polypeptides, polysaccharides, vitamins and the like, and the ginseng can dilate capillary vessels of the skin, promote blood circulation of the skin, enhance nutrition of the skin, and prevent the skin from dehydration, hardening and wrinkling, so that the elasticity of the skin is enhanced, and cells are regenerated. The Saviae Miltiorrhizae radix is rich in tanshinic acid and tanshinone, is a natural antioxidant, and has antiaging and anti-fibrosis effects. Radix Sophorae Flavescentis can balance oil secretion, dredge and astringe follicular orifice, and remove toxin and impurities in skin. The polysaccharide in the adenophora stricta has the radiation-resistant effect and the obvious protective effect on the skin damaged by radiation, and is also an excellent natural moisturizing factor. The radix scrophulariae extract has effects of reducing the content of inflammatory factors TNF-alpha, IL-1 beta and IL-6, increasing the concentration of anti-inflammatory factor IL-10, and realizing anti-inflammatory effect by influencing MAPK (mitogen activated protein kinase) pathway. The walnut kernel polyphenol extract in the walnut kernels not only has good effects of resisting oxidation, removing free radicals and resisting bacteria, but also has a certain inhibition effect on tyrosinase activity, and is a natural freckle-removing factor with great potential.

With the development of biotechnology, a variety of active enzymes have been isolated, identified, structurally interpreted, and studied for activity. Among them, cellulase has been widely studied and industrially used. In addition, the modern microbiological research results also bring new inspiration for the extraction of the traditional Chinese medicine. The method combines a novel traditional Chinese medicine active ingredient extraction technology formed by modern microbial engineering, obtains active ingredients in plants by utilizing a microbial fermentation technology, and develops a characteristic microbial source raw material into a new trend. During the growth process of microorganisms, a plurality of bioactive components including a plurality of enzymes, small molecular compounds and the like can be produced, and researches show that the substances are excellent skin care materials. Therefore, the whole skin care effect of the final product can be improved by the biological fermentation process.

According to the invention, high-pressure homogenization, enzymolysis reaction and microbial fermentation technology are organically combined, wherein the high-pressure homogenization and enzymolysis reaction can fully crush plant cells, so that the contents in the cells are quickly and fully released, the bioavailability of plant materials is improved, the subsequent fermentation operation is facilitated, the microbial fermentation can fully exert the biotransformation capability of microorganisms, the types and the contents of active ingredients in the five-ginseng fermentation product are effectively increased, and the antioxidation and anti-aging effects of the five-ginseng fermentation product are improved.

Preferably, the hydrolytic enzymes include cellulases and hemicellulases.

Preferably, the weight ratio of the five-ginseng stock solution to the cellulase to the hemicellulase is (1-500): (0.1-10): 0.1.

preferably, the fermentation strain comprises at least one of lactobacillus pentosus, lactobacillus plantarum, monascus and bifidobacterium, preferably lactobacillus pentosus, lactobacillus plantarum, monascus and bifidobacterium.

Preferably, the concentration of the fermentation strain is 10 ⁵ -10 ⁸ CFU/mL。

Specifically, the preparation method of the anti-oxidation and anti-aging five-ginseng fermented product comprises the following steps:

1) Mixing Ginseng radix, saviae Miltiorrhizae radix, radix Sophorae Flavescentis, radix scrophulariae, radix Adenophorae, semen Juglandis and deionized water, adding into a high pressure homogenizer, homogenizing at 20-40 deg.C under 40-100MPa for 3-30min to obtain stock solution of radix Ginseng;

preferably, the homogenizing temperature is 25-35 deg.C, homogenizing pressure is 40-100MPa, and homogenizing time is 3-15min;

2) Mixing five ginseng stock solution, cellulase and hemicellulase according to a mass ratio (1-500): (0.1-10): 0.1, adjusting pH to 4-7, carrying out enzymolysis at 40-60 deg.C under 40-60 rpm/min for 1-4 h, and heating at 80-120 deg.C for 10-40 min to obtain five-ginseng hydrolysate;

preferably, the pH value of enzymolysis is 5-6, the enzymolysis temperature is 45-55, the enzymolysis time is 1.5-3h, the heating temperature is 80-120 ℃, and the heating time is 10-40 min.

3) The five-ginseng hydrolysate, the fermentation strain, the MRS broth culture medium and deionized water are mixed according to the dosage ratio of (0.1-1000 g): (0.1-50 mL): (0.1-50 g): (0.1-1000 g), mixing, fermenting at 25-45 ℃ for 12-120 h, sterilizing the fermentation liquor at 100-121 ℃ for 20-30 min after fermentation is finished, then centrifuging for 20-30 min at the rotating speed of 4000-4500 rpm/min with the centrifugal radius of 15-18 cm, and taking the centrifuged supernatant to obtain the five-ginseng fermented product;

preferably, the amount ratio is (10-500 g): (1-10 mL): (1-10 g): (10-500 g), the fermentation temperature is 30-40 ℃, and the fermentation time is 12-96h.

The second aspect of the invention provides an anti-oxidation and anti-aging five-ginseng fermented product which is prepared by adopting the preparation method.

Preferably, the anti-oxidation and anti-aging five-ginseng fermentation product comprises the following components in concentration: ginsenoside Rb1 18.9-28.8 μ g/mL, salvianolic acid B311-386 μ g/mL, matrine 51-64 μ g/mL, and oxymatrine 52-76 μ g/mL.

The third aspect of the invention provides the application of the anti-oxidation and anti-aging five-ginseng fermentation product in cosmetics.

Specifically, the cosmetic comprises water aqua, emulsion, cream, spray, essence, soap or washing care.

Preferably, the essence comprises the antioxidant and anti-aging five-ginseng fermentation product and auxiliary materials.

Preferably, the anti-oxidation and anti-aging five-ginseng fermentation product accounts for 0.1-90% of the essence by mass.

Preferably, the auxiliary material comprises at least one of a humectant, an emollient and a preservative.

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

the invention effectively increases the types and the contents of active ingredients in the five-ginseng fermented product by organically combining high-pressure homogenization, enzymolysis reaction and microbial fermentation technologies, and particularly improves the contents of the active ingredients, wherein the ginsenoside Rb1 is 18.9-28.8 mu g/mL, the salvianolic acid B is 311-386 mu g/mL, the matrine is 51-64 mu g/mL, and the oxymatrine is 52-76 mu g/mL.

The five-ginseng fermentation product prepared by the invention has the hydroxyl radical clearance rate of 43.6-58% and the DPPH radical clearance rate of 50.19-69.96%, and has good antioxidant and anti-aging effects.

Detailed Description

In order to make the technical solutions of the present invention more apparent to those skilled in the art, the following examples are given for illustration. It should be noted that the following examples are not intended to limit the scope of the claimed invention.

The starting materials, reagents or apparatuses used in the following examples are conventionally commercially available or can be obtained by conventionally known methods, unless otherwise specified.

Example 1

A preparation method of a five-ginseng fermentation product comprises the following steps:

1) Mixing Ginseng radix 10g, saviae Miltiorrhizae radix 10g, radix Sophorae Flavescentis 10g, radix scrophulariae 10g, radix Adenophorae 10g, semen Juglandis 10g, and deionized water 540 g, adding into high pressure homogenizer at homogenizing temperature of 30 deg.C, homogenizing pressure of 80 MPa, and homogenizing for 4min to obtain stock solution of radix Ginseng;

2) Taking 300g of a five-ginseng stock solution, adding 0.9g of cellulase and 0.3g of hemicellulase, adjusting the pH to 5.5, carrying out enzymolysis on 2h at 50 ℃ and 50 rpm/min, and then heating at 100 ℃ for 20 min to obtain a five-ginseng hydrolysate;

3) 300g five-ginseng hydrolysate and 5mL lactobacillus pentosus bacterial liquid (with the concentration of 10) ⁷ CFU/mL), 5g of MRS broth culture medium and 200 g deionized water are mixed, fermentation is carried out at 37 ℃ for 72 h, the fermentation liquid is sterilized at 121 ℃ for 20 min after the fermentation is finished, then the fermentation liquid is centrifuged for 30min at the centrifugal radius of 15 cm and the rotating speed of 4000 rpm/min, and the centrifuged supernatant is taken, so that the five-ginseng fermented product is obtained.

Example 2

1) Mixing Ginseng radix 10g, saviae Miltiorrhizae radix 10g, radix Sophorae Flavescentis 10g, radix scrophulariae 10g, radix Adenophorae 10g, semen Juglandis 10g, and deionized water 540 g, adding into high pressure homogenizer at 25 deg.C and 40 MPa for 15min to obtain stock solution;

2) Taking 300g of a five-ginseng stock solution, adding 0.1g of cellulase and 0.1g of hemicellulase, adjusting the pH to 5, carrying out enzymolysis on 1.5 h at 45 ℃ and 50 rpm/min, and then heating at 110 ℃ for 15min to obtain a five-ginseng hydrolysate;

3) 300g five-ginseng hydrolysate and 1mL lactobacillus plantarum bacterial liquid (with the concentration of 10) ⁶ CFU/mL), 1g of MRS broth culture medium and 100 g deionized water are mixed, fermentation is carried out for 96h at 30 ℃, the fermentation broth is sterilized for 20 min at 121 ℃ after the fermentation is finished, then the fermentation broth is centrifuged for 30min at the centrifugal radius of 15 cm and the rotating speed of 4000 rpm/min, and the centrifuged supernatant is taken, thus obtaining the five-ginseng fermented product.

Example 3

1) Mixing Ginseng radix 10g, saviae Miltiorrhizae radix 10g, radix Sophorae Flavescentis 10g, radix scrophulariae 10g, radix Adenophorae 10g, semen Juglandis 10g, and deionized water 540 g, adding into a high pressure homogenizer, homogenizing at 35 deg.C under 100MPa for 3 min to obtain stock solution of radix Ginseng;

2) Taking 300g of a five-ginseng stock solution, adding 10g of cellulase and 10g of hemicellulase, adjusting the pH to 6, carrying out enzymolysis on 3h at 55 ℃ and 50 rpm/min, and then heating at 90 ℃ for 30min to obtain a five-ginseng hydrolysate;

3) Mixing 300g hydrolyzed solution of radix Ginseng Indici, 10mL Bacillus bifidus bacterial solution (with concentration of 10% ⁸ CFU/mL), 10g of MRS broth culture medium and 200 g deionized water are mixed, 50 h is fermented at 40 ℃, the fermentation broth is sterilized at 121 ℃ for 30min after the fermentation is finished, then the fermentation broth is centrifuged for 30min at the centrifugal radius of 15 cm and the rotating speed of 4000 rpm/min, and the centrifuged supernatant is taken, so that the five-ginseng fermented product is obtained.

Example 4

1) Mixing Ginseng radix 10g, saviae Miltiorrhizae radix 10g, radix Sophorae Flavescentis 10g, radix scrophulariae 10g, radix Adenophorae 10g, semen Juglandis 10g, and deionized water 540 g, adding into high pressure homogenizer, homogenizing at 20 deg.C under 100MPa for 15min to obtain stock solution of radix Ginseng;

3) 300g five-ginseng hydrolysate and 5mL Monascus ruber bacterial liquid (with the concentration of 10) ⁷ CFU/mL), 5g of MRS broth culture medium and 200 g deionized water are mixed, fermentation is carried out at 37 ℃ for 72 h, the fermentation liquid is sterilized at 121 ℃ for 20 min after the fermentation is finished, then the fermentation liquid is centrifuged for 30min at the centrifugal radius of 15 cm and the rotating speed of 4000 rpm/min, and the centrifuged supernatant is taken, so that the five-ginseng fermented product is obtained.

Example 5

This example is different from example 4 only in that the mixed bacterial liquid of Lactobacillus pentosus and Monascus is used as the monascus bacterial liquid in step 3), and the mixed bacterial liquid of Lactobacillus pentosus (10) ⁷ CFU/mL) and Monascus ruber liquid (10) ⁷ CFU/mL) ratio of 4:1, the amount of the mixed bacterial liquid is the same as that of the monascus bacterial liquid in example 4, and the other method steps are the same as those in example 4.

Example 6

This example is different from example 4 only in that the culture of Monascus purpureus bacteria in step 3) was replaced with a mixed culture of Lactobacillus pentosus and Monascus ruber bacteria, and that the culture of Lactobacillus pentosus (10) was contained in the mixed culture ⁷ CFU/mL) and Monascus ruber liquid (10) ⁷ CFU/mL) ratio of 1:1, the amount of the mixed bacterial liquid is the same as that of the monascus bacterial liquid in example 4, and the other method steps are the same as those in example 4.

Example 7

This example is different from example 4 only in that the mixed bacterial liquid of Lactobacillus pentosus and Monascus is used as the monascus bacterial liquid in step 3), and the mixed bacterial liquid of Lactobacillus pentosus (10) ⁷ CFU/mL) and Monascus ruber liquid (10) ⁷ CFU/mL) ratio of 1:4, the amount of the mixed bacterial liquid is the same as that of the monascus bacterial liquid in example 4, and the other method steps are the same as those in example 4.

Example 8

This example is different from example 4 only in that the mixed bacterial liquid of Lactobacillus plantarum and Monascus is used as the monascus bacterial liquid in step 3), and the mixed bacterial liquid contains Lactobacillus plantarum bacterial liquid (10) ⁷ CFU/mL) and Monascus ruber liquid (10) ⁷ CFU/mL) ratio of 4:1, the amount of the mixed bacterial liquid is the same as that of the monascus bacterial liquid in example 4, and the other method steps are the same as those in example 4.

Example 9

This example is different from example 4 only in that the mixed bacterial liquid of Lactobacillus plantarum and Monascus is used as the monascus bacterial liquid in step 3), and the mixed bacterial liquid contains Lactobacillus plantarum bacterial liquid (10) ⁷ CFU/mL) and Monascus ruber liquid (10) ⁷ CFU/mL) ratio of 1:1, the amount of the mixed bacterial liquid is the same as that of the monascus bacterial liquid in example 4, and the other method steps are the same as those in example 4.

Example 10

This example is different from example 4 only in that the mixed bacterial liquid of Lactobacillus plantarum and Monascus is used as the monascus bacterial liquid in step 3), and the mixed bacterial liquid isLactobacillus plantarum solution (10) ⁷ CFU/mL) and Monascus ruber liquid (10) ⁷ CFU/mL) ratio of 1:4, the amount of the mixed bacterial liquid is the same as that of the monascus bacterial liquid in example 4, and the other steps of the method are the same as those in example 4.

Example 11

This example is different from example 4 only in that the monascus liquid obtained in step 3) was replaced with a mixed liquid of bifidobacterium and monascus, and the bifidobacterium liquid (10) was contained in the mixed liquid ⁷ CFU/mL) and Monascus ruber liquid (10) ⁷ CFU/mL) ratio of 4:1, the amount of the mixed bacterial liquid is the same as that of the monascus bacterial liquid in example 4, and the other method steps are the same as those in example 4.

Example 12

This example is different from example 4 only in that the monascus liquid obtained in step 3) was replaced with a mixed liquid of bifidobacterium and monascus, and the bifidobacterium liquid (10) in the mixed liquid was used ⁷ CFU/mL) and Monascus ruber liquid (10) ⁷ CFU/mL) ratio of 1:1, the amount of the mixed bacterial liquid is the same as that of the monascus bacterial liquid in example 4, and the other method steps are the same as those in example 4.

Example 13

This example is different from example 4 only in that the monascus liquid obtained in step 3) was replaced with a mixed liquid of bifidobacterium and monascus, and the bifidobacterium liquid (10) in the mixed liquid was used ⁷ CFU/mL) and Monascus ruber liquid (10) ⁷ CFU/mL) ratio of 1:4, the amount of the mixed bacterial liquid is the same as that of the monascus bacterial liquid in example 4, and the other method steps are the same as those in example 4.

Example 14

This example differs from example 1 only in that the lactobacillus pentosus solution of step (3) is replaced with a mixed solution of lactobacillus pentosus and bifidobacterium, and that a lactobacillus pentosus solution (10) is present in the mixed solution ⁷ CFU/mL) and a bifidobacterium solution (10) ⁷ CFU/mL) ratio of 4:1, adding amount of mixed bacterial liquid and sugar milk in example 1The amount of the bacillus liquid added was the same as in example 1.

Example 15

This example differs from example 1 only in that the lactobacillus pentosus solution of step (3) is replaced with a mixed solution of lactobacillus pentosus and bifidobacterium, and that a lactobacillus pentosus solution (10) is present in the mixed solution ⁷ CFU/mL) and a bifidobacterium solution (10) ⁷ CFU/mL) ratio of 1:1, the amount of the mixed bacterial suspension was the same as that of the Lactobacillus saccharolyticus suspension of example 1, and the other examples were the same as example 1.

Example 16

This example is different from example 1 only in that the lactobacillus pentosus bacterial solution of step (3) is replaced with a mixed bacterial solution of lactobacillus pentosus and bifidobacterium, and a bacterial solution of lactobacillus pentosus (10) is present in the mixed bacterial solution ⁷ CFU/mL) and a bifidobacterium solution (10) ⁷ CFU/mL) ratio of 1:4, the amount of the mixed bacterial suspension was the same as that of the Lactobacillus saccharolyticus suspension of example 1, and the other examples were the same as example 1.

Example 17

This example is different from example 2 only in that the lactobacillus plantarum bacterial solution described in step (3) was replaced with a mixed bacterial solution of lactobacillus plantarum and bifidobacterium, and the lactobacillus plantarum bacterial solution (10) in the mixed bacterial solution ⁶ CFU/mL) and a bifidobacterium solution (10) ⁶ CFU/mL) ratio of 4:1, the amount of the mixed bacterial suspension was the same as that of the Bacillus plantarum bacterial suspension of example 2, and the other examples were the same as those of example 2.

Example 18

This example differs from example 2 only in that the lactobacillus plantarum solution described in step (3) was replaced with a mixed solution of lactobacillus plantarum and bifidobacterium, and a lactobacillus plantarum solution (10) in the mixed solution ⁶ CFU/mL) and a bifidobacterium solution (10) ⁶ CFU/mL) ratio of 1:1, the amount of the mixed bacterial suspension was the same as that of the Bacillus plantarum bacterial suspension of example 2, and the other examples were the same as those of example 2.

Example 19

This example differs from example 2 only in that the lactobacillus plantarum solution described in step (3) was replaced with a mixed solution of lactobacillus plantarum and bifidobacterium, and a lactobacillus plantarum solution (10) in the mixed solution ⁶ CFU/mL) and a bifidobacterium solution (10) ⁶ CFU/mL) ratio of 1: the amount of the mixed bacterial suspension was the same as that of the plant bacillus bacterial suspension of example 2, and the other examples were the same as example 2.

Example 20

3) 300g five ginseng hydrolysate and 1mL lactobacillus plantarum bacterial liquid (10) ⁷ CFU/mL), 3mL monascus liquid (10) ⁷ CFU/mL), 1mL bifidobacterium solution (10) ⁷ CFU/mL), 5g of MRS broth culture medium and 200 g deionized water are mixed, 84 h is fermented at 37 ℃, the fermentation broth is sterilized at 121 ℃ for 20 min after the fermentation is finished, then the fermentation broth is centrifuged for 30min at the centrifugal radius of 15 cm and the rotating speed of 4000 rpm/min, and the centrifuged supernatant is taken, so that the five-ginseng fermented product is obtained.

Example 21

1) Mixing Ginseng radix 10g, saviae Miltiorrhizae radix 10g, radix Sophorae Flavescentis 10g, radix scrophulariae 10g, radix Adenophorae 10g, semen Juglandis 10g, and deionized water 540 g, adding into high pressure homogenizer at homogenizing temperature of 30 deg.C, homogenizing pressure of 60 MPa, and homogenizing time of 6min to obtain stock solution of radix Ginseng;

3) 300g five-ginseng hydrolysate and 1mL lactobacillus pentosus bacterial liquid (10) ⁷ CFU/mL), 3mL monascus liquid (10) ⁷ CFU/mL), 1mL Bifidobacterium liquid (10 mL) ⁷ CFU/mL), 5g of MRS broth culture medium and 200 g deionized water are mixed, 84 h is fermented at 35 ℃, the fermentation broth is sterilized at 121 ℃ for 20 min after the fermentation is finished, then the fermentation broth is centrifuged for 30min at the centrifugal radius of 15 cm and the rotating speed of 4000 rpm/min, and the centrifuged supernatant is taken, so that the five-ginseng fermented product is obtained.

Example 22

2) Taking 300g of a five-ginseng stock solution, adding 0.3g of cellulase and 0.9g of hemicellulase, adjusting the pH to 5.5, carrying out enzymolysis on 2h at 50 ℃ and 50 rpm/min, and then heating at 100 ℃ for 20 min to obtain a five-ginseng hydrolysate;

3) 300g five-ginseng hydrolysate and 1mL lactobacillus pentosus bacterial liquid (10) ⁷ CFU/mL), 2mL monascus liquid (10) ⁷ CFU/mL), 1mL lactobacillus plantarum bacterial liquid (10) ⁷ CFU/mL), 5g of MRS broth culture medium and 200 g deionized water are mixed, 84 h is fermented at 35 ℃, the fermentation broth is sterilized at 121 ℃ for 20 min after the fermentation is finished, then the fermentation broth is centrifuged for 30min at the centrifugal radius of 15 cm and the rotating speed of 4000 rpm/min, and the centrifuged supernatant is taken, so that the five-ginseng fermented product is obtained.

Example 23

3) 300g five-ginseng hydrolysate and 5mL saccharomyces cerevisiae bacterial liquid (10) ⁷ CFU/mL), 5g YPD culture medium and 200 g deionized water, fermenting 72 h at 30 ℃, sterilizing the fermentation liquor at 121 ℃ for 20 min after the fermentation is finished, then centrifuging for 30min at the centrifugal radius of 15 cm and the rotating speed of 4000 rpm/min, and taking the centrifuged supernatant to obtain the five-ginseng fermented product.

Example 24

1) Mixing Ginseng radix 10g, saviae Miltiorrhizae radix 10g, radix Sophorae Flavescentis 10g, radix scrophulariae 10g, radix Adenophorae 10g, semen Juglandis 10g, and deionized water 540 g, adding into high pressure homogenizer at homogenizing temperature of 30 deg.C, homogenizing pressure of 80 MPa, and homogenizing time of 4min to obtain radix Ginseng stock solution;

2) Taking 300g of a five-ginseng stock solution, adding 0.9g of cellulase, adjusting the pH to 5.5, carrying out enzymolysis on 2h at 50 ℃ and 50 rpm/min, and then heating at 100 ℃ for 20 min to obtain a five-ginseng hydrolysate;

3) 300g hydrolyzed solution of radix Ginseng Indici, 5mL Monascus ruber bacterial solution (10) ⁷ CFU/mL), 5g of MRS broth culture medium and 200 g deionized water are mixed, 72 h is fermented at 30 ℃, the fermentation broth is sterilized at 121 ℃ for 20 min after the fermentation is finished, then the fermentation broth is centrifuged for 30min at the centrifugal radius of 15 cm and the rotating speed of 4000 rpm/min, and the centrifuged supernatant is taken, so that the five-ginseng fermented product is obtained.

Example 25

2) Taking 300g of the five-ginseng stock solution, adding 0.9g of hemicellulase, adjusting the pH to 5.5, carrying out enzymolysis on 2h at 50 ℃ and 50 rpm/min, and then heating at 100 ℃ for 20 min to obtain a five-ginseng hydrolysate;

3) 300g pentapanax hydrolyzed liquid, 5mL monascus fungus liquid (10) ⁷ CFU/mL), 5g of MRS broth culture medium and 200 g deionized water are mixed, 72 h is fermented at 30 ℃, the fermentation broth is sterilized at 121 ℃ for 20 min after the fermentation is finished, then the fermentation broth is centrifuged for 30min at the centrifugal radius of 15 cm and the rotating speed of 4000 rpm/min, and the centrifuged supernatant is taken, so that the five-ginseng fermented product is obtained.

Comparative example 1 (different from example 1 in that conventional water decoction is used)

mixing Ginseng radix 10g, saviae Miltiorrhizae radix 10g, radix Sophorae Flavescentis 10g, radix scrophulariae 10g, radix Adenophorae 10g, semen Juglandis 10g, and deionized water 540 g, heating the extract at 100 deg.C for 2 hr, filtering, and collecting filtrate.

Comparative example 2 (difference from example 1 in that no fermentation was carried out)

2) Taking 300g of the stock solution of the five-ginseng, adding 0.9g of cellulase and 0.3g of hemicellulase, adjusting the pH to 5.5, carrying out enzymolysis on 2h at 50 ℃ and 50 rpm/min, then heating for 20 min at 100 ℃, filtering, and collecting the filtrate.

Comparative example 3 (difference from example 1 in that high-pressure homogenization and fermentation were not conducted)

taking 10g of ginseng, 10g of salvia miltiorrhiza, 10g of radix sophorae flavescentis, 10g of radix scrophulariae, 10g of adenophora stricta and 10g of walnut kernel, adding 540 g of deionized water, 0.9g of cellulase and 0.3g of hemicellulase, adjusting the pH to 5.5, carrying out enzymolysis on 2h at 50 ℃ and 50 rpm/min, subsequently heating at 100 ℃ for 20 min, filtering and collecting filtrate.

Comparative example 4 (different from example 7 in that hydrolysis was not carried out)

2) Heating radix Ginseng stock solution 300g at 100 deg.C for 20 min, cooling to 30 deg.C, adding 1mL Lactobacillus pentosus liquid (10) ⁷ CFU/mL), 4mL monascus liquid (10) ⁷ CFU/mL), 5g of MRS broth culture medium and 200 g deionized water are mixed, 72 h is fermented at 37 ℃, after the fermentation is finished, the fermentation liquid is sterilized at 121 ℃ for 20 min, then the fermentation liquid is centrifuged for 30min at the centrifugal radius of 15 cm and the rotating speed of 4000 rpm/min, and the centrifuged supernatant is taken, so that the compound microbial inoculum is obtained.

Comparative example 5

Compared with example 4, the comparative example is different from example 4 only in that the bacterial liquid in step (3) is replaced by deionized water, and the addition amount is the same, and the other steps are also the same as example 4.

Test example 1: content determination of ginsenoside Rb1 and salvianolic acid B

The following tests were carried out on the samples prepared in examples 1 to 25 and comparative examples 1 to 5.

Sample preparation: taking 50g of sample solution, evaporating to dryness at 60 ℃ under reduced pressure, dissolving the residue with methanol, transferring to a 10mL measuring flask, adding methanol to dilute to scale, shaking, filtering, and taking the subsequent filtrate to obtain the sample solution.

Chromatographic conditions are as follows: octadecylsilane chemically bonded silica is used as a filling agent; acetonitrile is taken as a mobile phase A, and 0.1% phosphoric acid solution is taken as a mobile phase B; gradient elution was performed as specified in table 1 below; the column temperature is set to be 30 ℃, and the sample volume of the sample solution is 10 mu L; the detection wavelength of ginsenoside Rb1 is 205nm, and the detection wavelength of salvianolic acid B is 286nm.

The results of the content measurement are shown in Table 3.

TABLE 1

Test example 2: content determination of matrine and oxymatrine

Sample preparation: taking 20g of sample liquid, placing the sample liquid in a separating funnel, adding 0.4mL of concentrated ammonia test solution, adding trichloromethane for extraction, 25mL each time, combining the trichloromethane, recovering the solvent until the solvent is dry, adding a proper amount of methanol into residues for dissolving, transferring the residues into a 10mL measuring flask, adding methanol to the scale, and shaking up to obtain the product.

Chromatographic conditions are as follows: octadecylsilane chemically bonded silica is used as a filling agent; acetonitrile- [0.01mol/L ammonium acetate solution (PH 8.1 adjusted by concentrated ammonia solution) ] (3:2) is used as a mobile phase A, and 0.01mol/L ammonium acetate solution (PH 8.1 adjusted by concentrated ammonia solution) is used as a mobile phase B, and gradient elution is carried out according to the specification in the following table 2; the column temperature is set to be 30 ℃, and the sample volume of the sample solution is 10 mu L; the detection wavelength was 225nm.

The results of the content measurement are shown in Table 3.

TABLE 2

TABLE 3

Sample (I)	Ginsenoside Rb1 (mug/mL)	Salvianolic acid B (mug/mL)	Matrine (mu g/mL)	Oxymatrine (mug/mL)
					Example 1	20.1	311	51	54
Example 2	18.9	324	52	53
					Example 3	21.4	329	56	52
Example 4	21.0	316	55	54
					Example 5	25.5	375	59	63
Example 6	23.3	351	57	60
					Example 7	23.0	369	61	58
Example 8	25.4	336	60	62
					Example 9	23.1	335	58	61
Example 10	26.3	343	62	59
					Example 11	25.9	338	63	55
Example 12	21.8	361	57	58
					Example 13	23.3	344	59	60
Example 14	24.7	331	67	69
					Example 15	21.6	359	60	59
Example 16	22.1	336	68	54
					Example 17	25.4	335	63	66
Example 18	25.1	347	66	63
					Example 19	25.3	345	58	63
Example 20	26.3	333	60	61
					Example 21	25.5	349	67	62
Example 22	28.8	386	64	76
					Example 23	15.5	300	46	47
Example 24	17.4	308	49	46
					Example 25	16.8	303	48	49
Comparative example 1	6.9	112	21	18
					Comparative example 2	8.1	248	40	40
Comparative example 3	10.8	301	37	32
					Comparative example 4	9.4	273	39	41
Comparative example 5	11.7	294	44	45

As shown in Table 3, in examples 1 to 4, the content of ginsenoside Rb1 in the fermented product is 18.9 to 21.4. Mu.g/mL, the content of salvianolic acid B is 311 to 329. Mu.g/mL, the content of matrine is 51 to 56. Mu.g/mL, and the content of oxymatrine is 52 to 54. Mu.g/mL, by using single bacterial liquid for fermentation. Examples 5-19 are the co-fermentation of two bacterial solutions, wherein the ginsenoside Rb1 content in the fermentation product is 21.6-26.3. Mu.g/mL, the salvianolic acid B content is 335-375. Mu.g/mL, the matrine content is 57-68. Mu.g/mL, and the oxymatrine content is 55-69. Mu.g/mL. Examples 20 and 21 are the co-fermentation using three bacterial solutions, the ginsenoside Rb1 content in the fermentation product is 26.3 μ g/mL and 25.5 μ g/mL, the salvianolic acid B content is 333 μ g/mL and 349 μ g/mL, the matrine content is 60 μ g/mL and 67 μ g/mL, and the oxymatrine content is 61 μ g/mL and 62 μ g/mL. Example 22 is to use four kinds of bacteria liquid to ferment together, the content of ginsenoside Rb1 in the fermented product is 28.8 μ g/mL, the content of salvianolic acid B is 386 μ g/mL, the content of matrine is 64 μ g/mL, and the content of oxymatrine is 76 μ g/mL. Example 23 is a fermentation using Saccharomyces cerevisiae bacterial liquid, the content of ginsenoside Rb1 in the fermentation product is 15.5. Mu.g/mL, the content of salvianolic acid B is 300. Mu.g/mL, the content of matrine is 46. Mu.g/mL, and the content of oxymatrine is 47. Mu.g/mL. Examples 24-25 are hydrolysis with a single enzyme, and the fermentation product contains ginsenoside Rb1 16.8-17.4. Mu.g/mL, salvianolic acid B303-308. Mu.g/mL, matrine 48-49. Mu.g/mL, and oxymatrine 46-49. Mu.g/mL. The five ginseng fermentations prepared in examples 1-25, in which the active ingredient content was much higher than the samples prepared in comparative examples 1-5, demonstrated that different classes of natural active ingredients in the medicinal materials could be better released, especially ginsenoside with poor water solubility, by high pressure homogenization, hydrolysis and microbial fermentation. Example 22 co-fermentation with four bacterial solutions gave the highest active ingredient content and examples 23-25 gave the lowest active ingredient content. Examples 23 to 25 were also obtained by high-pressure homogenization, hydrolysis and fermentation, and the contents of each active ingredient in the fermented product were higher than those of the samples prepared in comparative examples 1 to 5, whereas example 23, which had been fermented with a saccharomyces cerevisiae solution, had each of examples 24 and 25 hydrolyzed with a single enzyme, resulting in the contents of each active ingredient in the fermented product of five ginsengs being lower than those of examples 1 to 22.

Test example 3: evaluation of erythrocyte hemolytic Properties

The erythrocyte hemolysis experiment is one of the alternatives of the rabbit eye irritation experiment (Draizetest), and the basic principle is to evaluate the damage of chemicals to eye tissue cells by measuring the dissolving amount and the denaturation degree of hemoglobin. The RBC test is used internationally for evaluating eye irritation studies on chemicals such as cosmetics and raw materials.

The samples prepared in examples 1 to 25 and comparative examples 1 to 5 were subjected to an erythrocyte hemolysis test according to the RBC test method and grade standard of the European Alternatives validation center (ECVAM). Wherein HD50 is the concentration of the sample at which 50% of the erythrocytes are hemolyzed, DI is the protein denaturation index, and L/D is the ratio of HD50 to DI. The evaluation criteria are shown in Table 4; the evaluation results are shown in Table 5.

TABLE 4

TABLE 5

As is clear from Table 5, the fermented products of the five ginsengs obtained in examples 1 to 25 and comparative examples 1 to 5 were not irritating to erythrocytes.

Test example 4: evaluation of hydroxyl radical scavenging ability

1.0 m LFeSO is added into the reaction system ₄ Solution (9.0 mmol/L), 1mL salicylic acid-ethanol solution (9.0 mmol/mL), 0.2 mL sample, 0.8 mL deionized water, and 1.0 mL H ₂ O ₂ (8.8 mmol/mL), reacting in 37 ℃ water bath for 30min, measuring the absorbance at 510 nm by using absolute ethyl alcohol as a blank control, and calculating the OH clearance (Y) of each extracting solution according to the following formula (1):

Y（%）=[1-(A ₁ -A ₂ )/A ₃ ]×100% (1)

in the formula A ₁ Is the absorbance of the reaction system of the extracting solution; a. The ₂ To add no H ₂ O ₂ Absorbance of the sample (absolute ethanol instead of H) ₂ O ₂ ）；A ₃ The absorbance of the blank control absolute ethyl alcohol is shown.

The results are shown in Table 6.

Test example 5: evaluation of DPPH radical scavenging ability

The ability to scavenge DPPH free radicals reflects to some extent the antioxidant capacity of the material. The greater the free radical clearance rate, the stronger the antioxidant capacity and the stronger the anti-aging capacity. Therefore, the anti-aging effect of the DPPH can be judged by researching the capability of the sample to remove DPPH free radicals. The following tests were carried out on the samples prepared in examples 1 to 25 and comparative examples 1 to 5.

Adding DPPH test solution 2.0 mL with a mass concentration of 45.8 mg/L and sample 0.05 mL into a test tube, supplementing the total volume to 3mL with 50% ethanol (v/v), shaking up, reacting in the dark for 30min, measuring the absorbance at 517 nm wavelength by using a 1 cm cuvette, and marking as A ₁ (ii) a Adding absolute ethanol 2mL and corresponding volume of sample to be measured into the test tube, supplementing the total volume to 3mL with 50% ethanol (v/v), and recording the measured absorbance as A ₂ (ii) a The DPPH test solution 2mL and 50% ethanol (v/v) 1mL were added to the tube, and the absorbance measured was recorded as A ₃ . Calculating the DPPH free radical clearance rate (Y) of the liquid to be detected according to the following formula (2);

the DPPH radical clearance is calculated as: y (%) = [1- (a) ₁ -A ₂ )/A ₃ ]×100 (2)

In the formula A ₁ Absorbance value of the System after eliminating DPPH for blank control, A ₂ Absorbance value of the System after removal of DPPH for the sample, A ₃ The absorbance value of the reaction system before adding the medicine is shown.

The results are shown in Table 6.

TABLE 6

Sample (I)	Hydroxyl radical scavenging rate (%)	DPPH radical scavenging ratio (%)	Sample (I)	Hydroxyl radical clearance (%)	DPPH radical scavenging ratio (%)
						Example 1	45.04	50.19	Example 16	50.18	65.12
Example 2	44.50	54.38	Example 17	53.02	59.08
						Example 3	47.34	52.09	Example 18	49.55	58.13
Example 4	43.60	55.30	Example 19	49.47	60.33
						Example 5	51.33	67.79	Example 20	51.80	56.97
Example 6	48.65	63.01	Example 21	55.22	58.43
						Example 7	48.23	56.68	Example 22	58.0	69.96
Example 8	55.12	60.03	Example 23	38.82	45.11
						Example 9	48.07	63.66	Example 24	45.45	48.32
Example 10	49.36	62.22	Example 25	43.29	49.08
						Example 11	56.19	59.54	Comparative example 1	17.69	15.06
Example 12	54.77	66.36	Comparative example 2	26.77	30.18
						Example 13	50.06	63.21	Comparative example 3	33.09	39.84
Example 14	47.76	63.44	Comparative example 4	36.18	44.79
						Example 15	48.10	57.29	Comparative example 5	28.99	32.78

As shown in Table 6, in examples 1 to 4, the hydroxyl radical scavenging rate was 43.60 to 47.34% and the DPPH radical scavenging rate was 50.19 to 55.30% by using the single bacterial liquid fermentation. In examples 5-19, two bacterial solutions were used for co-fermentation, the hydroxyl radical clearance rate was 47.76-56.19%, and the DPPH radical clearance rate was 56.68-67.79%. Examples 20 and 21 were co-fermented with three bacterial solutions, hydroxyl radical scavenging rates of 51.80% and 55.22%, and DPPH radical scavenging rates of 56.97% and 58.43%. In example 22, the hydroxyl radical clearance rate is 58.0% and the DPPH radical clearance rate is 69.96% for co-fermentation with four bacterial solutions. In example 23, the hydroxyl radical scavenging rate of the fermentation is 38.82% and the DPPH radical scavenging rate is 45.11% by adopting the saccharomyces cerevisiae bacterial liquid for fermentation. Examples 24-25, which were hydrolyzed using a single enzyme, had hydroxyl radical clearance of 43.29-45.45% and DPPH radical clearance of 48.32-49.08%. The hydroxyl radical scavenging rate and DPPH radical scavenging rate of the fermented schizandra sinensis prepared in examples 1 to 25 were much higher than those of the samples prepared in comparative examples 1 to 5, wherein the hydroxyl radical scavenging rate and DPPH radical scavenging rate were the best in example 22, and the hydroxyl radical scavenging rate and DPPH radical scavenging rate were lower in examples 23 to 25.

Test example 6: evaluation of tyrosinase inhibitory Activity

The ability to inhibit tyrosinase activity reflects the whitening ability of the material to a certain extent, and the greater the tyrosinase activity inhibition rate, the greater the whitening ability, and the samples prepared in examples 1 to 25 and comparative examples 1 to 5 were tested.

Accurately sucking a sample solution, a Phosphate Buffer Solution (PBS) with the pH value of 6.8 and a dopa solution with the mass fraction of 0.1% according to the table 7, fully mixing, incubating at 30 ℃ for 5min, respectively adding tyrosinase solutions (the activity is 100U/mL) with corresponding volumes, incubating at 30 ℃ for 10 min, quickly transferring to a cuvette, and measuring the absorbance at 475 nm.

TABLE 7

Calculating the relative inhibition rate (I) of the sample solution to tyrosinase according to the formula (3).

I＝[(A ₁ -A ₂ )-(A ₃ -A ₄ )] /(A ₁ -A ₂ ) ×100% （3）

In the formula, A ₁ 、A ₂ 、A ₃ And A ₄ The absorbance of each of the solutions of groups 1-4.

The results are shown in Table 8.

TABLE 8

As can be seen from table 8, the five ginseng fermentations prepared in examples 1-25 have a very good inhibitory effect on tyrosinase activity, and show stronger whitening efficacy than the samples prepared in comparative examples 1-5. Comparative examples 1 and 5 are those which had not been subjected to microbial fermentation and had very weak tyrosinase inhibitory activity. It can be seen that the activity is significantly improved after the fermentation treatment, and particularly, the fermented product prepared by co-fermenting the four bacterial liquids in example 22 has the highest inhibition on the tyrosinase activity.

Test example 7: haCaT cell viability after UV irradiation

HaCaT (human immortalized keratinocytes) cells were seeded in 6-well plates at 37 ℃ with 5% CO ₂ The incubator of (2) incubate overnight. When the cell plating rate in the 6-well plate reaches 50% -60%, adding a cell culture medium into a blank control group and a negative control group, adding a cell culture medium containing 5% of a test sample (5%) with a corresponding concentration into the test sample group, pretreating for 1h, and then performing UVB irradiation (30 mJ/cm) ² ) And (4) 40min. After further culturing at 37 ℃ in an incubator containing 5% CO2 for 24 hours, the medium was aspirated, washed 2 to 3 times with PBS, and the cell viability was measured by the MTT method, the results of which are shown in Table 9.

TABLE 9

As can be seen from table 9, the five ginseng fermentations prepared in examples 1-25 have better protective effects on HaCat cells after UV irradiation and can promote the growth of cells, compared to comparative examples 1-5. In this respect, the fermented product prepared by fermenting the mixed bacterial liquid is superior to the fermented product prepared by fermenting the single bacterial liquid.

Test example 8: activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in human dermal fibroblasts (HSD) after UV irradiation, MDA, MMP-1 and COL-I content

Cells were seeded into 6-well plates with a mass fraction of 5% CO at 37 ℃ ₂ Incubate overnight. When the cell plating rate in the 6-well plate reaches 60% -70%, adding cell culture medium into the blank control group and the negative control group, adding cell culture medium containing 5% of test substance with corresponding concentration into the test sample group for pretreatment of 1h, and then carrying out UVB irradiation (30 mJ/cm) ² ) And (4) 40min. At 37 ℃ and 5 mass percent of CO ₂ The incubator continues to culture 24 h. After the culture is finished, the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) and the content of Malondialdehyde (MDA) are measured according to the operation method of the kit. The contents of matrix metalloproteinase-1 (MMP-1) and human type I collagen (COL-I) were determined according to the ELISA kit protocol.

Watch 10

Sample (I)	SOD (U/mg protein)	GSH-Px (U/mg protein)	MDA content (μ g/mg protein)	MMP-1 content (ng/mL)	COL-I content (ng/mL)
						Example 1	54.68	366	15.45	6.54	109
Example 2	56.75	351	15.39	6.32	112
						Example 3	52.09	363	15.72	6.71	120
Example 4	53.14	371	15.11	6.83	116
						Example 5	70.10	430	15.47	5.39	125
Example 6	64.31	403	14.88	6.02	127
						Example 7	63.48	411	15.30	5.88	120
Example 8	66.77	425	15.76	5.63	141
						Example 9	60.05	379	14.96	6.20	126
Example 10	62.38	396	15.43	5.21	124
						Example 11	58.73	448	14.31	6.04	128
Example 12	62.19	415	14.20	6.02	124
						Example 13	65.87	392	15.16	5.77	122
Example 14	67.08	394	15.64	6.37	133
						Example 15	58.79	378	15.39	6.16	126
Example 16	64.43	403	15.25	5.09	123
						Example 17	58.68	465	13.99	6.18	140
Example 18	59.59	413	14.17	5.69	126
						Example 19	69.12	437	14.86	5.46	121
Example 20	68.28	404	14.16	5.87	137
						Example 21	63.33	417	14.37	6.06	126
Example 22	70.91	450	13.04	5.11	147
						Example 23	46.63	344	16.62	6.93	105
Example 24	50.03	323	16.23	6.99	102
						Example 25	48.83	337	16.85	7.01	106
Comparative example 1	19.88	142	22.17	9.11	52.4
						Comparative example 2	37.24	360	19.71	7.55	93.1
Comparative example 3	21.70	206	16.88	7.08	103
						Comparative example 4	45.79	311	17.54	6.92	96.5
Comparative example 5	34.97	243	20.76	9.73	72.4

The results in Table 10 show that the five-ginseng fermentations in experimental examples 1-25 can improve the activities of SOD and GSH-Px in HSD, reduce the content of MDA and show stronger antioxidant activity than the samples prepared in comparative examples 1-5. In addition, the five ginseng fermentates prepared in experimental examples 1-25 can reduce MMP-1 content and increase COL-I content more than the samples prepared in comparative examples 1-5. The increase of MMP-1 and the decrease of COL-I can cause skin aging and wrinkle, and the results show that the five-ginseng leavening in the experimental examples 1-25 has stronger anti-aging effect. In the aspects of antioxidation and anti-aging effects, the fermented product prepared by fermenting the mixed bacterial liquid is superior to the fermented product prepared by fermenting the single bacterial liquid.

Application example 1: and evaluating the whitening effect of the essence.

The present application example provides essence solutions prepared from the fermented five-ginseng products obtained in example 4 or comparative example 1, respectively, and the specific formulations thereof are shown in table 11.

TABLE 11

The evaluation target of the effect selects 30 volunteers, the age of which is 20-45 years old, and the male and female halves. The skin care essence prepared in example 4 was applied to the left face, and the skin care essence prepared in comparative example 1 was applied to the right face, and the face was applied once a day, each time 1g product was applied to the face, and massaged for 2 minutes. Filling out a trial evaluation table according to the sensory evaluation. The composition is used for 7 days, 14 days and 28 days, and counted once. The volunteers evaluate the whiteness (0-5 min) and compactness (0-5 min) effects of the samples in the using process, wherein the highest score is 5, which shows that the effect is obvious, and the lowest score is 0, which shows that the effect is not good. The results are shown in Table 12.

TABLE 12

Application example 2: and evaluating the wrinkle-reducing efficacy of the essence.

The subject for evaluation of the effects was 30 volunteers, and women aged 30-45 years were selected. Before the test, pictures were taken using a VISIA facial image analyzer to analyze the fine lines of the volunteers' eyes. The volunteers were asked to use the corresponding group of products after cleaning the face, and after 1 minute, the images were again taken and analyzed as the instant light streaking effect of the product. Then, the volunteers were asked to continuously try out a sample to be tested (essence prepared from the five-ginseng fermented product obtained in example 4 or comparative example 1) 2 times a day, 0.5 g each time, continuously try out for 28d, make a return visit on days 7 and 28, take a photograph of the face and analyze it as the long-lasting light streak effect of the product. The detection of the same volunteer is completed by the same detection person. And calculating the change rate of the fine lines of the eyes by using an analysis system arranged in the detection instrument, and counting the result. The results are shown in Table 13.

Watch 13

As can be seen from tables 12 and 13, the volunteers considered that the skin whiteness, firmness and soothing were significantly improved after using the essence prepared in example 4, which is superior to that of the essence prepared in comparative example 1, and the effect is related to the ability of the five ginseng fermentates to scavenge free radicals and increase intracellular active enzymes; the essence prepared in example 4 can improve fine lines of eyes, and the effect is related to that the five-ginseng fermented product can improve the content of COL-I, and is also superior to that of the essence prepared in comparative example 1. The application of the five-ginseng fermentation product in cosmetics has obvious antioxidant and anti-aging effects.

While the preferred embodiments of the present invention have been illustrated and described, it will be understood by those skilled in the art that the present invention is not limited to the details of the embodiments shown and described, but is capable of numerous equivalents and substitutions without departing from the spirit of the invention as set forth in the claims appended hereto.

Claims

1. A preparation method of a five-ginseng fermentation product is characterized by comprising the following steps:

1) Homogenizing the five-ginseng raw material to obtain a five-ginseng stock solution; the five ginseng raw materials comprise ginseng, salvia miltiorrhiza, radix sophorae flavescentis, adenophora stricta, radix scrophulariae and walnut kernels; the homogenizing pressure is 40-100MPa;

3) Fermenting the five-ginseng hydrolysate by utilizing a fermentation strain to obtain a five-ginseng fermented product;

the hydrolytic enzyme consists of cellulase and hemicellulase;

the fermentation strain consists of lactobacillus pentosus, lactobacillus plantarum, monascus and bifidobacteria.

2. The preparation method according to claim 1, wherein the weight ratio of the five-ginseng stock solution to the cellulase to the hemicellulase is (1-500): (0.1-10): 0.1.

3. the method according to claim 1, wherein the concentration of the fermentation strain is 10 ⁵ -10 ⁸ CFU/mL。

4. A five-ginseng fermented product, which is prepared by the preparation method of any one of claims 1 to 3, and comprises the following components in concentration: ginsenoside Rb1 18.9-28.8 μ g/mL, salvianolic acid B311-386 μ g/mL, matrine 51-64 μ g/mL, and oxymatrine 52-76 μ g/mL.

5. Use of the five-ginseng fermented product according to claim 4 in the preparation of cosmetics.

6. An essence, characterized by comprising the five-ginseng fermented product of claim 4 and an auxiliary material.

7. The essence of claim 6, wherein the five-ginseng fermentation product accounts for 0.1-90% of the essence by mass.