CN114632055B

CN114632055B - Whitening and relieving plant fermentation product and preparation method and application thereof

Info

Publication number: CN114632055B
Application number: CN202210266922.1A
Authority: CN
Inventors: 何敬愉; 潘丹阳; 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: 2022-03-17
Filing date: 2022-03-17
Publication date: 2023-03-21
Anticipated expiration: 2042-03-17
Also published as: CN114632055A

Abstract

The invention belongs to the technical field of cosmetics, and discloses a whitening and relieving plant fermentation product and a preparation method and application thereof. The preparation method of the plant fermentation product comprises the following steps: homogenizing the plant raw material under high pressure to obtain plant stock solution; mixing the plant stock solution, the fermentation strain, melibiose and oligomannose, and fermenting to obtain a plant fermentation product; the plant material comprises one or more of fructus Anisi Stellati, flos Matricariae Chamomillae, and Sargassum; the fermentation strain comprises one or more of lactobacillus salivarius, lactobacillus plantarum and lactobacillus casei. According to the invention, high-pressure homogenization and directional microbial fermentation technologies are organically combined, the biotransformation capability of microbes is fully exerted, the obtained plant fermentation product can effectively inhibit the activity of tyrosinase, the content of inflammatory factors IL-1 beta and TNF-alpha generated by HaCat cells after UV irradiation is reduced, the skin whitening and relieving effects are remarkable, and the irritation of phenethyl resorcinol in cosmetics can be effectively reduced.

Description

Whitening and relieving plant fermentation product and preparation method and application thereof

Technical Field

The invention belongs to the technical field of cosmetics, and particularly relates to a whitening and soothing plant fermentation product as well as a preparation method and application thereof.

Background

With the continuous development of science, the components and the effects of natural plants are continuously excavated, and the plant extracts are more and more widely applied to the fields of medicine, food and daily chemicals. The development of the times also leads people to pay more and more attention to natural components and green beauty, and promotes the plant extract to be widely applied to beauty and skin care cosmetics such as whitening, freckle removing, sun protection, repairing and the like. Although plant extracts obtained by conventional extraction techniques (such as water extraction) have been applied to cosmetics, the conventional extraction techniques cannot effectively extract active ingredients from plants, and the extracts have limited active ingredients and cannot sufficiently exert the efficacy of plants in cosmetics.

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 plant fermentation product with the whitening and relieving effects is provided by the invention, and the preparation method and the application thereof are characterized in that the plant fermentation product with the whitening and relieving effects is prepared by organically combining high-pressure homogenization and directional microbial fermentation technologies.

The first aspect of the present invention provides a method for preparing a plant fermentation product, comprising the steps of:

homogenizing the plant raw material under high pressure to obtain plant stock solution;

mixing the plant stock solution, the fermentation strain, melibiose and oligomannose, and fermenting to obtain the plant fermentation product;

wherein the pressure of the high-pressure homogenization is 40-100MPa;

the plant material comprises one or more of anise, chamomile and seaweed;

the fermentation strain comprises one or more of lactobacillus salivarius, lactobacillus plantarum and lactobacillus casei.

Compared with the traditional extraction technology, the modern microbial fermentation technology also brings new inspiration for plant extraction. By combining with the plant active ingredient extraction technology formed by modern microbial engineering, a characteristic plant-microbial source raw material can be developed. Research shows that microorganisms can produce a plurality of bioactive components including a plurality of enzymes, small molecular compounds and the like in the growth process, and the bioactive components are excellent skin care materials. After the plants are subjected to microbial fermentation treatment, the product of the plant-based biological fermentation treatment has a more excellent and more effective skin protection effect; the products are compounded and applied by a scientific method, so that the skin protection function can be effectively strengthened, skin nutrition is supplemented, the self-regulating capacity of the skin is improved, the beauty cosmetics are more suitable for human body cells, the microcirculation of skin tissues is improved, the activity of the skin tissue cells is enhanced, the metabolism of the skin tissue cells is promoted, and the skin is in a healthy state. One or more of the sea fennel, the chamomile and the seaweed are used as plant raw materials, and the high-pressure homogenization technology and the directional microbial fermentation technology are organically combined, wherein the high-pressure homogenization can fully crush plant cells, so that the contents in the cells are quickly and fully released, the biological utilization efficiency of the plant materials is improved, and the subsequent fermentation operation is facilitated; meanwhile, the biotransformation capability of the microorganisms can be fully exerted through specific microorganism fermentation substrates (melibiose and oligomannose) and fermentation strains, and the obtained plant fermentation product has remarkable whitening and relieving effects.

Preferably, the preparation method of the plant fermentation product further comprises the following steps:

after the fermentation is finished, sterilizing the fermentation liquor at 100-121 ℃ for 20-30min, then centrifuging for 20-30min at the centrifugal radius of 15-25cm and the rotating speed of 4000-8000rpm/min, and taking the centrifuged supernatant to obtain the plant fermentation product.

Preferably, the plant material comprises anise, chamomile and seaweed.

Preferably, the fermentation strain is one of lactobacillus salivarius, lactobacillus plantarum or lactobacillus casei.

Preferably, the temperature of the high-pressure homogenization is 25-50 ℃, and the time of the high-pressure homogenization is 3-30min. More preferably, the temperature of the high-pressure homogenization is 30-45 ℃, and the time of the high-pressure homogenization is 5-15min. Most preferably, the temperature of the high-pressure homogenization is 35 ℃, the pressure of the high-pressure homogenization is 60-80MPa, and the time of the high-pressure homogenization is 10-15min.

Preferably, MRS broth and water are also added during the mixing.

Preferably, the plant stock solution, the fermentation strain, the MRS broth, the melibiose, the oligomannose and the water are used in a ratio of (0.1-1000 g): (0.1-50 mL): (0.1-50 g): (0.01-50 g): 1g: (0.1-1000 g). More preferably, the plant stock solution, the fermentation strain, the MRS broth, the melibiose, the oligomannose and the water are used in an amount ratio of (10-500 g): (1-10 mL): (1-10 g): (0.1-5 g): 1g: (10-500 g). Most preferably, the plant stock solution, the fermentation strain, the MRS broth, the melibiose, the oligomannose and the water are used in a ratio of (10-100 g): (1-5 mL): (1-5 g): (0.1-3 g): 1g: (10-100 g).

Preference is given toThe concentration of the fermentation strain is 10 ⁵ -10 ⁹ CFU/mL。

Preferably, the temperature of the fermentation is 30-45 ℃, and the time of the fermentation is 12-120h. More preferably, the temperature of the fermentation is 35-40 ℃, and the time of the fermentation is 12-96h.

In a second aspect, the invention provides a plant fermentation product, which is prepared by the preparation method of the invention.

In a third aspect, the invention provides the use of the plant fermentation product of the invention in cosmetics.

Preferably, the plant fermentation product is applied to reducing the irritation of phenethyl resorcinol in cosmetics.

Based on the application, the invention also provides a cosmetic, which comprises the plant fermentation product and an acceptable auxiliary material in the cosmetic, wherein the plant fermentation product accounts for 0.01-80% of the cosmetic by mass.

Preferably, the cosmetically acceptable adjuvant comprises one or more of an emulsifier, a humectant, a thickener, and a preservative.

Preferably, the plant fermentation product accounts for 0.05-20% of the cosmetic by mass.

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

according to the invention, high-pressure homogenization and directional microorganism fermentation technologies are organically combined, and meanwhile, the biotransformation capability of microorganisms can be fully exerted through a specific microorganism fermentation substrate (melibiose and oligomannose) and a fermentation strain, the inhibition rate of the obtained plant fermentation product on the activity of in vitro tyrosinase is 46.3-68.4%, the relative activity of intracellular tyrosinase is 33.8-49.3%, the contents of inflammatory factors IL-1 beta and TNF-alpha generated by HaCat cells after UV irradiation can be reduced, the anti-inflammatory activity is shown, and therefore, the skin-whitening and soothing effects are remarkable; the plant fermentation product prepared by the invention can also effectively reduce the irritation of phenylethyl resorcinol in cosmetics.

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 plant fermentation product comprises the following steps:

1) Mixing 30g of hypecoum vulgare and 270g of deionized water, and adding into a high-pressure homogenizer, wherein the homogenization temperature is 35 ℃, the homogenization pressure is 60MPa, and the homogenization time is 12min, so as to obtain a plant stock solution;

2) 100g of the stock solution of the plant and 3mL of the Lactobacillus plantarum bacterial solution (concentration: 10) ⁷ CFU/mL), 2g of MRS broth culture medium, 1g of melibiose, 1g of oligomannose and 100g of deionized water are mixed, fermentation is carried out for 72h at 37 ℃, the fermentation broth is sterilized at 121 ℃ for 20min after the fermentation is finished, then centrifugation is carried out for 30min at the centrifugation radius of 15cm and the rotation speed of 5000rpm/min, and the centrifuged supernatant is taken to obtain the plant fermentation product.

Example 2

1) Mixing 30g of chamomile and 270g of deionized water, and adding into a high-pressure homogenizer, wherein the homogenizing temperature is 35 ℃, the homogenizing pressure is 80MPa, and the homogenizing time is 10min to obtain a plant stock solution;

2) 100g of the stock solution of the plant and 3mL of the Lactobacillus plantarum bacterial solution (concentration: 10) ⁷ CFU/mL), 2g of MRS broth culture medium, 0.8g of melibiose, 1.2g of oligomannose and 100g of deionized water are mixed, fermentation is carried out for 72h at 37 ℃, the fermentation liquor is sterilized for 20min at 121 ℃ after the fermentation is finished, then centrifugation is carried out for 25min under the conditions that the centrifugation radius is 15cm and the rotation speed is 5000rpm/min, and the centrifuged supernatant is taken to obtain the plant fermentation product.

Example 3

1) Mixing 30g of seaweed and 270g of deionized water, and then adding the mixture into a high-pressure homogenizer, wherein the homogenization temperature is 35 ℃, the homogenization pressure is 80MPa, and the homogenization time is 15min, so as to obtain a plant stock solution;

2) 100g of the stock solution of the plant and 3mL of the Lactobacillus plantarum bacterial solution (concentration: 10) ⁷ CFU/mL), 1g of MRS broth culture medium, 1.2g of melibiose, 0.8g of oligomannose and 100g of deionized water are mixed, fermentation is carried out for 72 hours at 37 ℃, the fermentation liquor is sterilized for 20 minutes at 121 ℃ after the fermentation is finished, then centrifugation is carried out for 30 minutes at the centrifugal radius of 15cm and the rotating speed of 5000rpm/min, and the centrifuged supernatant is taken to obtain the plant fermentation product.

Example 4

1) Mixing 18g of sea fennel, 12g of chamomile and 270g of deionized water, and then adding the mixture into a high-pressure homogenizer, wherein the homogenization temperature is 35 ℃, the homogenization pressure is 70MPa, and the homogenization time is 15min, so as to obtain a plant stock solution;

2) 100g of the stock solution of the plant and 3mL of the Lactobacillus plantarum bacterial solution (concentration: 10) ⁷ CFU/mL), 2g of MRS broth culture medium, 0.8g of melibiose, 1.2g of oligomannose and 100g of deionized water are mixed, fermentation is carried out for 72h at 37 ℃, the fermentation liquor is sterilized for 20min at 121 ℃ after the fermentation is finished, then centrifugation is carried out for 30min at the centrifugal radius of 15cm and the rotating speed of 5000rpm/min, and the centrifuged supernatant is taken to obtain the plant fermentation product.

Example 5

1) Mixing 24g of sea fennel, 6g of seaweed and 270g of deionized water, and then adding the mixture into a high-pressure homogenizer, wherein the homogenization temperature is 35 ℃, the homogenization pressure is 70MPa, and the homogenization time is 12min, thus obtaining a plant stock solution;

2) 100g of the stock solution of the plant and 3mL of the Lactobacillus plantarum bacterial solution (concentration: 10) ⁷ CFU/mL), 2g of MRS broth culture medium, 0.8g of melibiose, 1.2g of oligomannose and 100g of deionized water, fermenting for 72 hours at 37 ℃, sterilizing the fermentation broth at 121 ℃ for 20 minutes after the fermentation is finished, centrifuging for 30 minutes at the centrifugal radius of 15cm and the rotating speed of 5000rpm/min, and taking outAnd (4) obtaining the plant fermentation product from the supernatant after the heart.

Example 6

1) Mixing 18g of sea fennel, 6g of chamomile, 6g of seaweed and 270g of deionized water, and then adding the mixture into a high-pressure homogenizer, wherein the homogenization temperature is 35 ℃, the homogenization pressure is 80MPa, and the homogenization time is 15min, so as to obtain a plant stock solution;

2) 100g of the stock solution of the plant and 3mL of the Lactobacillus plantarum bacterial solution (concentration: 10) ⁷ CFU/mL), 2g of MRS broth culture medium, 0.8g of melibiose, 1.2g of oligomannose and 100g of deionized water are mixed, fermentation is carried out for 72 hours at 37 ℃, the fermentation broth is sterilized for 20 minutes at 121 ℃ after the fermentation is finished, then centrifugation is carried out for 30 minutes at the centrifugation radius of 15cm and the rotation speed of 5000rpm/min, and the centrifuged supernatant is taken to obtain the plant fermentation product.

Example 7

Compared with the example 1, the difference is only that the lactobacillus plantarum bacterial liquid obtained in the step 2) is replaced by the lactobacillus salivarius bacterial liquid, the addition amount of the bacterial liquid is the same as that of the lactobacillus plantarum bacterial liquid obtained in the example 1, and other components, addition amounts and preparation methods are the same as those of the example 1.

Example 8

Compared with the example 2, the difference is only that the lactobacillus plantarum bacterial liquid obtained in the step 2) is replaced by the lactobacillus salivarius bacterial liquid, the addition amount of the bacterial liquid is the same as that of the lactobacillus plantarum bacterial liquid obtained in the example 2, and other components, addition amounts and preparation methods are the same as those of the example 2.

Example 9

Compared with the embodiment 3, the difference is only that the lactobacillus plantarum bacterial liquid obtained in the step 2) is replaced by a lactobacillus salivarius bacterial liquid, the addition amount of the bacterial liquid is the same as that of the lactobacillus plantarum bacterial liquid obtained in the embodiment 3, and other components, addition amounts and preparation methods are the same as those of the embodiment 3.

Example 10

Compared with the example 4, the difference is only that the lactobacillus plantarum bacterial liquid obtained in the step 2) is replaced by the lactobacillus salivarius bacterial liquid, the addition amount of the bacterial liquid is the same as that of the lactobacillus plantarum bacterial liquid obtained in the example 1, and other components, addition amounts and preparation methods are the same as those of the example 4.

Example 11

Compared with the example 5, the difference is only that the lactobacillus plantarum bacterial liquid obtained in the step 2) is replaced by the lactobacillus salivarius bacterial liquid, the addition amount of the bacterial liquid is the same as that of the lactobacillus plantarum bacterial liquid obtained in the example 1, and other components, addition amounts and preparation methods are the same as those of the example 5.

Example 12

Compared with the embodiment 6, the difference is only that the lactobacillus plantarum bacterial liquid obtained in the step 2) is replaced by a lactobacillus salivarius bacterial liquid, the addition amount of the bacterial liquid is the same as that of the lactobacillus plantarum bacterial liquid obtained in the embodiment 1, and other components, addition amounts and preparation methods are the same as those of the embodiment 6.

Example 13

Compared with the example 1, the difference is that the lactobacillus plantarum bacterial liquid obtained in the step 2) is replaced by lactobacillus casei bacterial liquid, the addition amount of the bacterial liquid is the same as that of the lactobacillus plantarum bacterial liquid obtained in the example 1, and other components, addition amounts and preparation methods are the same as those of the example 1.

Example 14

Compared with the example 2, the difference is that the lactobacillus plantarum bacterial liquid obtained in the step 2) is replaced by lactobacillus casei bacterial liquid, the addition amount of the bacterial liquid is the same as that of the lactobacillus plantarum bacterial liquid obtained in the example 2, and other components, addition amounts and preparation methods are the same as those of the example 2.

Example 15

Compared with the embodiment 3, the difference is only that the lactobacillus plantarum bacterial liquid obtained in the step 2) is replaced by lactobacillus casei bacterial liquid, the addition amount of the bacterial liquid is the same as that of the lactobacillus plantarum bacterial liquid obtained in the embodiment 3, and other components, addition amounts and preparation methods are the same as those of the embodiment 3.

Example 16

Compared with the example 4, the difference is only that the lactobacillus plantarum bacterial liquid obtained in the step 2) is replaced by lactobacillus casei bacterial liquid, the addition amount of the bacterial liquid is the same as that of the lactobacillus plantarum bacterial liquid obtained in the example 1, and other components, addition amounts and preparation methods are the same as those of the example 4.

Example 17

Compared with the embodiment 5, the difference is only that the lactobacillus plantarum bacterial liquid obtained in the step 2) is replaced by lactobacillus casei bacterial liquid, the amount of the bacterial liquid added is the same as that of the lactobacillus plantarum bacterial liquid obtained in the embodiment 1, and other components, addition amounts and preparation methods are the same as those of the embodiment 5.

Example 18

Compared with the embodiment 6, the difference is only that the lactobacillus plantarum bacterial liquid obtained in the step 2) is replaced by lactobacillus casei bacterial liquid, the amount of the bacterial liquid added is the same as that of the lactobacillus plantarum bacterial liquid obtained in the embodiment 1, and other components, addition amounts and preparation methods are the same as those of the embodiment 6.

Comparative example 1 (differing from example 6 only in that conventional water decoction was used)

A method for preparing plant extract comprises the following steps:

mixing fructus Anisi Stellati 18g, flos Matricariae Chamomillae 6g, sargassum 6g, and deionized water 540g, heating and extracting at 100 deg.C for 2 hr, filtering, and collecting filtrate to obtain plant extract.

Comparative example 2 (the only difference from example 6 is that the Lactobacillus plantarum solution was replaced with deionized water)

Compared with the example 6, the comparative example only differs from the example 6 in that the lactobacillus plantarum bacterial liquid obtained in the step 2) is replaced by deionized water, the addition amount is the same, and other components, addition amounts and preparation methods are the same as those of the example 6.

Comparative example 3 (differing from example 6 only in that no melibiose and no oligomannose were added)

This comparative example is compared with example 6, except that no melibiose and oligomannose were added in step 2), and the other components, addition amounts, and preparation methods were the same as example 6.

Comparative example 4 (differing from example 6 only in that fructose was substituted for melibiose)

This comparative example is compared with example 6, except that melibiose was replaced with fructose in step 2) and the amounts added were the same, and the other components, the amounts added, and the preparation method were the same as in example 6.

Comparative example 5 (differing from example 6 only in that oligomannose was replaced with inulin)

This comparative example is compared with example 6, except that the oligomannose was replaced with inulin in step 2), and the added amount was the same, and the other components, added amount, and preparation method were the same as example 6.

Comparative example 6 (only different from example 6 in that the Lactobacillus plantarum bacterial solution was replaced with the Saccharomyces cerevisiae bacterial solution)

Compared with the example 6, the comparative example only differs from the example 6 in that the lactobacillus plantarum bacterial liquid obtained in the step 2) is replaced by the saccharomyces cerevisiae bacterial liquid, the addition amount is the same, and other components, addition amounts and preparation methods are the same as those of the example 6.

Test example 1: 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 samples prepared in examples 1 to 18 and comparative examples 1 to 6 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 a 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 1; the evaluation results are shown in Table 2.

TABLE 1

L/D	Grading
		L/D>100	Has no irritation
10<L/D≤100	Micro-stimulation property
		1<L/D≤10	Mild irritation
0.1<L/D≤1	Irritation due to poisoning
		L/D≤0.1	Severe irritation property

TABLE 2

As can be seen from Table 2, the samples prepared in examples 1 to 18 and comparative examples 1 to 6 were not irritating to erythrocytes.

Test example 2: evaluation of tyrosinase Activity inhibition in vitro

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 18 and comparative examples 1 to 6 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 3, fully mixing, incubating at 30 ℃ for 5min, respectively adding tyrosinase solutions (the activity is 100U/mL) with corresponding volumes, incubating at 30 ℃ for 10min, quickly transferring to a cuvette, and measuring the absorbance at 475 nm.

TABLE 3

Group of	Group 1	2 groups of	Group 3	4 groups of
					Sample solution (mL)	0	0	1.0	1.0
Phosphate buffer solution (mL)	2.0	2.5	1.0	1.5
					Dopa solution (mL)	1.0	1.0	1.0	1.0
Tyrosinase solution (mL)	0.5	0	0.5	0
					Total volume (mL)	3.5	3.5	3.5	3.5

Calculating the relative inhibition rate (I) of the sample solution to tyrosinase according to the formula (1).

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

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

The test results are shown in Table 4.

TABLE 4

Test example 3: intracellular tyrosinase Activity evaluation

Inoculation of B16 melanoma cells into 6-well plates, at 37 ℃ and 5% CO ₂ The incubator of (a) was incubated overnight. The cells were then incubated with the sample (4%) for 48 hours. Cells were then washed three times with ice-cold PBS, lysed with 1% polyethylene glycol octylphenyl ether (TritonX-100)/PBS, and centrifuged at 13000rpm for 20 minutes. The supernatant was then subjected to protein quantification using a bicinchoninic acid (BCA) assay kit. After a solution of potassium phosphate buffer (100mM, pH 6.8), cell lysate (50. Mu.g) and 10mM L-DOPA (DOPA) was placed at 37 ℃ for incubation for 1h, absorbance was measured at 475 nm. The test results are shown in table 5.

Test example 4: intracellular tyrosinase activity

Inoculation of B16 melanoma cells into 6-well plates, at 37 ℃ and 5% CO ₂ The incubator of (a) was incubated overnight. The cells were then incubated with the sample (4%) for 48 hours. After treatment, cells were washed three times with ice-cold PBS, lysed with 1% TritonX-100/PBS, and centrifuged at 13000rpm for 20 minutes. The small eye-associated transcription factor (MITF) in the supernatant was measured using an enzyme-linked immunosorbent assay (ELISA) kit. The test results are shown in table 5.

TABLE 5

As is clear from a combination of the test results in tables 4 and 5, the fermented plant products prepared in examples 1 to 18 had a superior inhibitory effect on tyrosinase activities in vitro and in cells, compared to the samples prepared in comparative examples 1 to 6; the plant fermentations prepared in examples 1-18 may reduce the amount of MITF, a key protein for melanogenesis. The plant fermented products of examples 1 to 18 showed stronger whitening efficacy in combination of the three indexes. Comparative example 1 and comparative example 2 are those which have not been fermented by microorganisms and have weak inhibitory activity against tyrosinase, and it can be seen that the inhibitory activity of the present invention is significantly improved after fermentation treatment. Comparative example 3 does not add melibiose and oligomannose, and the inhibitory activity to tyrosinase is lower than that of examples 1-18, which shows that the two components of melibiose and oligomannose are added for co-fermentation, so that the inhibitory activity to tyrosinase can be remarkably improved, and the content of MITF is reduced.

Test example 5: effect on HaCaT cell viability after UV irradiation

Seeding of HaCaT cells into 6-well plates, at 37 ℃ and 5% CO ₂ The incubator of (a) was incubated overnight. When the cell plating rate in the 6-well plate reaches 50% -60%, adding a cell culture medium into the blank control group and the negative control group, and adding a test sample groupThe cell culture medium containing the corresponding concentration of the test sample (4%) was added, pretreated for 1h and then subjected to UVB irradiation (30 mJ/cm) ² ) And (4) 40min. CO at 37 ℃ and 5% ₂ The culture box of (1) was further cultured for 24 hours, and then the medium was aspirated, washed with PBS 2 to 3 times, and the cell viability was tested by MTT method, the test results are shown in Table 6.

TABLE 6

The results in Table 6 show that the plant fermentations prepared in examples 1-18 have a better protective effect on HaCat cells after UV irradiation and are able to promote cell growth than comparative examples 1-6.

Test example 6: effect on IL-1 beta and TNF-alpha content in HaCaT cells after UV irradiation

Seeding of HaCaT cells into 6-well plates, at 37 ℃ and 5% CO ₂ The incubator of (a) was incubated 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 a test sample (4%) with a corresponding concentration into a test sample group, pretreating for 1h, and then performing UVB irradiation (30 mJ/cm) ² ) And (4) 40min. CO at 37 ℃ and 5% ₂ The cultivation was continued for 24h. After the culture, the contents of IL-1. Beta. And TNF-. Alpha. Were determined according to the ELISA kit protocol, and the results are shown in Table 7.

TABLE 7

The results in Table 7 show that the plant fermentations prepared in examples 1-18 can reduce the levels of the inflammatory factors IL-1. Beta. And TNF-. Alpha. Produced by HaCat cells after UV irradiation, and exhibit anti-inflammatory activity, as compared to comparative examples 1-6.

Application example 1

An essence comprising the plant fermentation product prepared in example 6 of the present invention. The specific formulation is shown in table 8. The preparation method comprises the steps of preparing the essence according to a conventional preparation method.

TABLE 8

Composition (A)	Mass fraction (%)
		Deionized water	Balance of
Glycerol	12.00
		1,3 butanediol	6.00
Pentanediol	3.00
		EDTA disodium salt	0.02
Carbomer	0.15
		Beta-glucan	4.00
Example 6 preparation of plant fermentation	2.00
		1,2 hexanediol	0.80
Phenylethylated Resorcinol	0.50
		P-hydroxyacetophenone	0.40

Comparative application example 1

The essence of this comparative application example 1 is different from the essence of application example 1 only in that the plant fermented product prepared in example 6 is replaced with the plant fermented product prepared in comparative example 3, and the other components, the addition amounts and the preparation method are the same as those of application example 1.

Comparative application example 2

The essence of this comparative application example 2 is different from the essence of application example 1 only in that the plant fermentation product prepared in example 6 was replaced with the plant fermentation product prepared in comparative example 4, and the other components, the addition amounts and the preparation method were the same as those of application example 1.

Comparative application example 3

The essence of this comparative application example 3 is different from the essence of application example 1 only in that the plant fermentation product prepared in example 6 was replaced with the plant fermentation product prepared in comparative example 5, and the other components, the addition amounts and the preparation method were the same as those of application example 1.

Comparative application example 4

The essence of this comparative application example 4 is different from the essence of application example 1 only in that the plant fermentation product prepared in example 6 was replaced with the plant fermentation product prepared in comparative example 6, and the other components, the addition amounts and the preparation method were the same as those of application example 1.

Blank control group

The essence of the blank control group is different from that of application example 1 only in that the plant fermentation product prepared in example 6 is replaced by deionized water, and other components, addition amounts and preparation methods are the same as those of application example 1.

The whitening and soothing efficacy evaluation was performed on the essences of the application example 1, the comparative application examples 1 to 4, and the blank group.

The subject selected 60 volunteers, aged 20-45 years, male and female halves. The essence liquids of application example 1, comparative application examples 1-4 and blank group were used respectively for 6 groups, and were applied once a day, each time 1g of the 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 evaluated the whiteness and soothing effects of the samples during use, with a maximum score of 5 indicating significant effect and a minimum score of 0 indicating no effect. The results are shown in Table 9.

TABLE 9

As can be seen from table 9, the volunteers considered that the skin whiteness and soothing were significantly improved using the essence prepared in application example 1, which is superior to using the essences of comparative application examples 1 to 4 and the blank group. The whitening effect is related to the tyrosinase activity inhibition ability of the plant fermentation product; the soothing effect is associated with the inhibition of inflammatory factor production by the plant fermentation. In addition, the phenethyl resorcinol has irritation at a certain concentration, and compared with a blank control group, the application example 1 has a relieving score obviously higher than that of the blank control group, so that the plant fermentation product disclosed by the invention can well reduce the irritation of the phenethyl resorcinol.

Application example 2

An emulsion comprising the plant ferment prepared in example 6 of the present invention. The specific formulation is shown in table 10. The preparation method is prepared according to the conventional preparation method of the emulsion.

Watch 10

Composition (I)	Mass fraction (%)
		Deionized water	Balance of
Octyl dodecanol	20.00
		Undecylenoyl phenylalanine	10.00
Bisabolol	3.00
		PEG-40 sodium stearate	2.80
Ascorbyl tetraisopalmitate	1.00
		Phenethylresorcinol	0.50
Example 6 preparation of plant fermentation	2.00
		1,2 hexanediol	0.80
P-hydroxyacetophenone	0.40

Comparative application example 5

The emulsion of this comparative application example 5 differs from the emulsion of application example 2 only in that the plant fermentation product prepared in example 6 was replaced with the plant fermentation product prepared in comparative example 3, and the other components, the amounts to be added, and the preparation method were the same as those of application example 2.

Comparative application example 6

The emulsion of this comparative application example 6 is different from the emulsion of application example 2 only in that the plant fermentation product prepared in example 6 is replaced with the plant fermentation product prepared in comparative example 4, and the other components, the addition amounts and the preparation method are the same as those of application example 2.

Comparative application example 7

The emulsion of this comparative application example 7 is different from the emulsion of application example 2 only in that the plant fermentation product prepared in example 6 was replaced with the plant fermentation product prepared in comparative example 5, and the other components, the addition amounts and the preparation method were the same as those of application example 2.

Comparative application example 8

The emulsion of this comparative application example 8 is different from the emulsion of application example 2 only in that the plant fermentation product prepared in example 6 was replaced with the plant fermentation product prepared in comparative example 6, and the other components, the addition amounts and the preparation method were the same as those of application example 2.

Blank control group

The emulsion of the blank control group is different from that of application example 2 only in that the plant fermentation product prepared in example 6 is replaced by deionized water, and other components, addition amounts and preparation methods are the same as those of application example 2.

The whitening and soothing efficacy evaluation was performed on the emulsions of application example 2, comparative application examples 5-8, and the blank group.

The subject selected 60 volunteers, aged 20-45 years, male and female halves. The emulsions of application example 2, comparative application examples 5 to 8 and blank were used for the respective groups of 6. The tested part is the face, and after the face of the testee is cleaned in the morning and evening every day, a proper amount of the product is uniformly applied to the face by pressing and is massaged until the product is absorbed. It is administered twice daily, in the morning and evening. Subjects were followed before (day 0) and 14 and 28 days after product use. The evaluation indices included skin erythema and desquamation at the test site, and each index was evaluated in terms of its degree from 0 point (very good) to 9 points (very poor). In addition, the repair and soothing efficacy of the product was evaluated by comparing the skin moisture content, skin TEWL and skin redness area before and after use of the product. The results are shown in Table 11.

TABLE 11

As can be seen from Table 11, the subjects considered that the skin erythema and desquamation were improved by using the emulsion of application example 2, as compared with the emulsions of comparative application examples 5 to 8 and the blank group; the water content of the horny layer, the TEWL value and the red area change before and after the use of the test subject are combined, and the plant leavening has remarkable relieving and repairing functions.

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 method for preparing a plant fermentation product is characterized by comprising the following steps:

wherein the pressure of the high-pressure homogenization is 40-100MPa;

the plant raw material is composed of one or more of anise, chamomile and seaweed;

the fermentation strain consists of one or more of lactobacillus salivarius, lactobacillus plantarum and lactobacillus casei.

2. The method according to claim 1, wherein the temperature of the high-pressure homogenization is 25-50 ℃, and the time of the high-pressure homogenization is 3-30min.

3. The method of claim 1, wherein the mixing further comprises adding MRS broth and water.

4. The method according to claim 3, wherein the plant stock solution, the fermentation strain, the MRS broth, the melibiose, the oligomannose and the water are used in a ratio of (0.1-1000 g): (0.1-50 mL): (0.1-50 g): (0.01-50 g): 1g: (0.1-1000 g).

5. The method according to claim 1, wherein the concentration of the fermentation strain is 10 ⁵ -10 ⁹ CFU/mL。

6. The method according to claim 1, wherein the fermentation temperature is 30-45 ℃ and the fermentation time is 12-120h.

7. A plant fermentation product, which is produced by the production method according to any one of claims 1 to 6.

8. Use of the plant fermentation product of claim 7 for the preparation of cosmetics.

9. Use of the plant ferment of claim 7 for reducing the irritativeness of phenethyl resorcinol in cosmetics.

10. The cosmetic is characterized by comprising the plant fermentation product of claim 7 and cosmetically acceptable auxiliary materials, wherein the plant fermentation product accounts for 0.1-80% of the mass of the cosmetic.