KR102654331B1 - Preparation and application of a fermentation composition with anti-photoaging,calming,cooling effects - Google Patents

Abstract

The present invention discloses a fermented composition having anti-photoaging, soothing and cooling effects, as well as its manufacturing method and application. The fermentation composition consists of a mass ratio of cephalic acid: peach blossom: soothing composition = 20 to 50: 0.5 to 5: 50 to 100, and the soothing composition is peach resin: cactus: dendrobium in a ratio of 3:1:1 or 3.5:2:2. Included in the ratio of The fermentation composition provided by the present invention can prevent photoaging damage, especially photoaging damage caused by both the amount of heat contained in light and the conversion of light energy into heat energy, and at the same time, the composition removes free radicals and inflammatory factors. By suppressing the emission of , indirect damage caused by solar radiation to the skin can be reduced. The fermentation method used in the present invention avoids the loss of active ingredients due to the traditional extraction process, protects the skin in various ways through the synergistic effect of the composition, improves the anti-photoaging effect, and reduces skin dryness, roughness, and darkening. , can improve aging symptoms.

Description

Fermentation composition with anti-photoaging, soothing and cooling effects and its manufacturing method and application {Preparation and application of a fermentation composition with anti-photoaging, calming, cooling effects}

The present invention applies to the field of cosmetics, and specifically relates to a fermented composition having anti-photoaging, soothing and cooling effects, and its production method and application.

Sunscreen refers to a substance added to cosmetics to protect the skin from damage caused by specific ultraviolet rays or to protect the product itself by absorbing, reflecting or scattering light. As consumer awareness and interest in UV protection increases, UV protection products are no longer limited to specific situations such as outdoors or in summer or only in terms of UV protection, and the public is placing more importance on sun protection in all aspects.

However, even products with a high SPF value still do not provide 100% protection, and for example, the blue light damage that modern people face and the injuries caused by highly penetrating ultraviolet radiation, sunscreens in UV blocking products also have a very good prevention effect. I can't do it. In addition, although sunscreens can absorb, reflect and refract ultraviolet rays, they are helpless against the amount of heat generated by light irradiation. In the case of organic sunscreens, some sunscreens mainly absorb UVB and some mainly absorb UVA. Some are broad-spectrum sunscreens, meaning they can absorb UVB and UVA. These types of sunscreens can also convert the light energy of absorbed UV rays into heat energy.

Photoaging is a complex process that causes characteristic deformation of the skin due to excessive sunlight irradiation, mainly manifesting as skin dryness, sagging, wrinkles, increase or loss of pigment, and capillary dilatation. Damage caused by photoaging mainly includes DNA damage, mitochondrial damage, peroxidation of intracellular biological macromolecules (proteins, phospholipids, etc.) and degradation of extracellular matrix (collagen, elastic fibers), inflammatory cascade, and immune suppression. In addition to photoaging that can be caused by ultraviolet rays, an increasing number of studies are showing that visible light (VL) and infrared (IR) light can also cause photoaging. For example, large amounts of reactive oxygen species (ROS) can induce an oxidative stress response and promote the expression of matrix metalloproteinase (MMP)-1 and MMP-9, causing the infiltration of inflammatory cells and It can cause skin aging by causing the decomposition of matrix collagen, and pigmentation induced by VL irradiation causes skin darkening and aging. In addition, the increase in skin temperature caused by light irradiation and the free radicals and heat energy generated during the energy conversion process of sunscreen also further promote skin photoaging.

Advanced glycation end products (AGEs) are brown macromolecular products that are not easily decomposed, produced by non-enzymatic glycation reactions between free amino acids and reducing sugars of macromolecular substances such as proteins, lipids, and nucleic acids. Research results have shown that AGEs accumulate in large quantities in photoaged skin and play an important role in various stages of photoaging. However, AGEs can be phagocytosed by cells such as macrophages and fibroblasts and then decomposed and removed by lysosomes or ubiquitin-proteasomes.

Duchong is a famous medicinal tree seed native to China. It contains various effective active ingredients, mainly compounds such as flavonoids, chlorogenic acids, iridoids, polysaccharides, polyphenols, phenylpropanoids, and lignans. It can be used for antioxidant, free radical removal, anti-aging, promotion of organism function, anti-cancer, etc. As a result of the study, it was found that eucalyptus has the effect of blocking sunlight and preventing damage caused to the skin by light irradiation. First, eucalyptus has an anti-infrared effect and reduces the amount of heat that infrared rays apply to the skin. Next, Eucommia regulates temperature-sensitive pathways such as TRPV4 and produces a cooling effect on the skin by stimulating temperature-sensitive pathways. The transient receptor potential (TRP) channel is a non-voltage-dependent cation channel that is widely expressed in various tissues of mammals, such as the skin and oral cavity, and is involved in various important physiological functions of the organism. TRPV1, TRPV4, TRPM8, etc. are sensitive to temperature. Research has shown that some Chinese herbal medicine ingredients act to regulate the function of TRPM8, such as rhubarb and yellow lotus can lower the body temperature of mice with yeast fever, and such functions are related to TRPV1 and TRPV1. It was found that it may be involved in effective regulation of TRPM8. In addition, active ingredients such as aucubin contained in eucalyptus can achieve a temperature reduction effect by dilating blood vessels and increasing caloric dissipation.

Peach blossom is the flower of the peach tree, a deciduous tree in the Rosaceae family. Its beauty and health-supporting effects have been widely applied in traditional Chinese medicine. In Shennong Bencao Jing, the first pharmaceutical book, it was mentioned that peach blossoms have the effect of “improving a person’s complexion”; In <Dogyeong Herbal Medicine>, it is recorded that “it moisturizes a person’s face and improves the complexion through moisturizing,” and the beauty method of Medicine King Son Sa-mak also highly evaluates the effects of peach blossom. Modern medical and nutritional research has also revealed that peach blossoms are rich in chemical components such as polyphenols, polysaccharides, carotenoids, various vitamins, amino acids, and trace elements, and have relatively excellent medicinal and nutritional values.

In the prior art, UV blocking products only protect against UV rays and take care of the photoaging phenomenon caused by UV rays, but also indirectly damage the skin due to irradiation of light, such as the amount of heat directly caused by irradiation and the energy generated during the energy conversion process of the sunscreen. There was a failure to systematically care for the phenomenon that promotes photoaging of the skin in various aspects, such as the amount of heat generated, thermal damage caused to the skin through direct and indirect pathways, and damage due to oxidative stress caused to the skin by VL and IR. The goal is to solve the problem. The fermented composition can cool the skin, prevent thermal aging of the skin, and prevent skin photoaging caused by VL and IR. In addition, ultrasonic extraction can improve extraction efficiency, rupture lactic acid bacilli, and release ingredients beneficial to the skin. It removes substances such as cell walls that cause skin antagonism and increases affinity with the skin. As the use of sunscreen products becomes more widespread and the duration of use becomes longer, the fermented composition has the effect of soothing sensitivity and relieving irritation, while simultaneously reducing the discomfort caused by tanned skin. The fermented composition can not only fundamentally prevent photoaging of the skin, but also soothe tanned skin, reduce the accumulation of AGEs, brighten skin tone, and reduce degeneration of elastic tissue caused by sunlight. By using fresh plants, low-temperature drying and fermentation methods, more active ingredients beneficial to the skin can be retained and the utilization rate of raw materials can be increased.

The present application is implemented with the following technical solutions:

One aspect of the present invention provides a fermentation composition having anti-photoaging, soothing and cooling effects, which is obtained by grinding, fermenting and extracting eucommia, peach blossom and a soothing composition having a mass ratio of 20 to 50:0.5 to 5:50 to 100. Obtained by a method for producing the fermentation composition;

The soothing composition is a combination of peach resin, cactus and dendrobium; The mass ratio of the composition is peach resin, cactus and dendrobium 3:1:1 or 3.5:2:2; The soothing composition prepared as above is used.

Preferably, the mass ratio of the cephalic acid:peach blossom:sedative composition is 30 to 40:1 to 3:60 to 90.

More preferably, the mass ratio of the cephalic acid:peach blossom:sedative composition is 35:2:75 or 30:2:80.

Preferably, the manufacturing method is specifically as follows:

1) Sorting and washing the eucalyptus, peach blossom, and soothing composition, drying at low temperature, and then weighing them according to ratio;

2) pulverizing peach resin, cactus, and dendrobium, adding them to deionized water, immersing them, and stirring for 2 hours;

3) pulverizing the peach blossoms and peach blossoms, adding them to 2) and performing wet ultra-fine grinding to obtain a homogenate;

4) inoculating the homogenate obtained in 3) with the bacterial solution and fermenting it while stirring at a uniform speed;

5) After fermentation is complete, ultrasonic extraction, filtration and sterilization are performed, and the filtrate is centrifuged to obtain a supernatant;

6) filtering the supernatant obtained in 5) sequentially using microfiltration membranes having different pore sizes, collecting the filtrate, and obtaining the fermentation composition,

More preferably:

The low-temperature drying conditions in step 1) are drying below 50°C; The drying method includes, but is not limited to, one or a combination of oven drying, freeze drying, and vacuum drying;

The volume of deionized water in step 2) is 5 to 20 times the total mass of eucalyptus, peach blossom and soothing composition;

In step 3), the wet ultrafine grinding is performed using a colloid mill or homogenizer.

More preferably:

In step 4), the inoculating bacterial solution includes, but is not limited to, one or a combination of lactic acid bacilli, lactic acid bacteria, and yeast;

In step 4), the inoculum inoculation amount is 5% to 10% of the mass of the homogenate, the bacterial content is ≥1.0Х10 ¹¹ cfu/g, the fermentation temperature is 28 to 35°C, and the fermentation time is 12 to 48 hours.

More preferably:

The pore size range of the microfiltration membrane in step 6) is 0.1 to 10 μm, preferably 10 μm, 0.45 μm, and 0.2 μm.

In some more specific embodiments, the fermentation composition is prepared by the following process:

(1) Sorting and washing the eucalypts, peach blossoms, and soothing composition, drying them at low temperature, and then weighing them according to proportions;

(2) pulverizing peach resin, cactus, and dendrobium, then adding 20 times the total mass of deionized water of the eucalyptus, peach blossom, and soothing composition weighed in step 1), immersing them, and stirring for 2 hours;

(3) pulverizing the peach blossoms and peach blossoms, adding them to 2) and performing ultra-fine grinding using a wet method to obtain a homogenate;

(4) A step in which 10% of the mass of the homogenate is inoculated into the homogenate obtained in 3) and fermented with stirring at 35°C for 48 hours. The bacterial solution is lactic acid bacillus, and the bacterial content of the bacterial solution is ≥1.0Х10 11 cfu. /g;

(5) After fermentation is complete, ultrasonic extraction is performed for 20 minutes, filtered, sterilized, and the filtrate is centrifuged to obtain a supernatant;

(6) Filtering the supernatant obtained in 5) sequentially using microfiltration membranes with pore sizes of 10 μm, 0.45 μm, and 0.2 μm, and collecting the filtrate to obtain the fermentation composition.

The drying method in step 1) is freeze drying;

In step 3), the wet ultrafine grinding is performed using a homogenizer.

The mass ratio of the peach resin, cactus and dendrobium is 3.5:2:2.

Another aspect of the invention provides a method for preparing a fermentation composition comprising the following steps:

1) Sorting and washing the eucalyptus, peach blossom, and soothing composition, drying at low temperature, and then weighing them according to ratio;

2) pulverizing peach resin, cactus, and dendrobium, adding them to deionized water, immersing them, and stirring for 2 hours;

3) pulverizing the peach blossoms and peach blossoms, adding them to 2) and performing wet ultra-fine grinding to obtain a homogenate;

4) inoculating the homogenate obtained in 3) with the bacterial solution and fermenting it while stirring at a uniform speed;

5) After fermentation is complete, ultrasonic extraction, filtration and sterilization are performed, and the filtrate is centrifuged to obtain a supernatant;

6) filtering the supernatant obtained in 5) using microfiltration membranes having different pore sizes sequentially and collecting the filtrate to obtain the fermentation composition;

The low-temperature drying conditions in step 1) are drying below 50°C; The drying method includes, but is not limited to, one or a combination of oven drying, freeze drying, and vacuum drying;

The volume of deionized water in step 2) is 5 to 20 times the total mass of eucalyptus, peach blossom and soothing composition;

In step 3), the wet ultrafine grinding is performed using a colloid mill or homogenizer;

In step 4), the inoculating bacterial solution includes one or a combination of two of lactic acid bacilli, lactic acid bacteria, and yeast, but is not limited thereto; The inoculant inoculum amount is 5% to 10% of the homogenate mass, the bacterial content is ≥1.0Х10 ¹¹ cfu/g, the fermentation temperature is 28 to 35°C, and the fermentation time is 12 to 48 hours;

The pore size range of the microfiltration membrane in step 6) is 0.1 to 10 μm, preferably 10 μm, 0.45 μm, and 0.2 μm.

Preferably, the mass ratio of the soothing composition composed of eucalyptus, peach blossom, and peach resin, cactus and dendrobium is 30 to 40:1 to 3:60 to 90;

The mass ratio of the peach resin, cactus and dendrobium is 3:1:1 or 3.5:2:2.

Another aspect of the present invention provides a cosmetic product containing the fermentation composition; The cosmetics include, but are not limited to, liquid, emulsion, spray, spray, cream, mask pack, and liquid foundation.

Compared with the prior art, the present invention can achieve the following technical effects:

When the total concentration is the same, the DPPH removal rate of the fermentation composition provided by the present application is higher than the removal rate of a single component, which explains that a better skin antioxidant effect can be obtained through the combination of the three raw materials. Additionally, the present invention uses water as a solvent, avoids the introduction of organic reagents, uses fresh plants as raw materials, and retains more active ingredients through a low-temperature drying method and fermentation process. In addition, by removing components such as cell walls and membranes after fermentation, affinity with the skin can be increased and irritation that may occur when used as a cosmetic raw material can be reduced. Compared to existing methods, the extraction temperature is low and the efficiency is high.

In addition, peach resin, cactus, and dendrobium contained in the soothing composition of the present application are all rich in polysaccharide components, so they have a very strong skin moisturizing effect and can prevent damage to the skin barrier. As a result of the study, it was discovered that the soothing composition has a very strong anti-allergy and anti-irritant effect and can relieve allergy or irritation symptoms caused by cosmetics or other elements on the skin.

Figure 1 shows the inhibition rate (%) of MMP-1 expression after using the fermentation composition in Experimental Example 3.
Figure 2 shows the inhibition rate (%) of AGEs production after using the fermentation composition in Experimental Example 4.
Figure 3 shows the skin elasticity recovery ratio (R2) values before and after using the fermented composition in Experimental Example 6.
Figure 4 is a graph of the percentage reduction in transepidermal water loss (TEWL) values after using the fermentation composition in Example 6.
Figure 5 is a graph showing skin texture before and after using the fermented composition in Experimental Example 6.

Below, the implementation method of the present application will be described in detail through examples, so that the implementation process of how to apply the technical means in the present application to achieve the technical task is fully understood and implemented accordingly.

Unless otherwise specified, all raw materials and equipment used in this institute are those commonly used in the industry and are all commercially purchased products. Unless otherwise specified, all methods used herein are conventional methods in the art.

There are various other technical solutions that can be implemented in the present application, but they are not listed one by one in this specification, and all technical solutions for which protection is claimed in the claims of the present application can also be implemented.

Example

In the examples and comparative examples of the present invention described below, a fermentation composition having anti-photoaging, soothing and cooling effects was prepared by the following process:

(1) Sorting and washing the eucalyptus, peach blossom, and soothing composition, drying at low temperature, and then weighing them according to ratio;

(2) pulverizing peach resin, cactus, and dendrobium, then adding 20 times the total mass of deionized water of the eucalyptus, peach blossom, and soothing composition weighed in step 1), immersing them, and stirring for 2 hours;

(3) pulverizing the peach blossoms and peach blossoms, adding them to 2) and performing wet ultra-fine grinding to obtain a homogenate;

(4) A step of inoculating the homogenate obtained in 3) with 10% of the mass of the homogenate and fermenting it at 35°C with stirring for 48 hours. The bacterial solution is lactic acid bacilli, and the bacterial content of the bacterial solution is ≥1.0×10 ¹¹ cfu/g;

(5) After fermentation is complete, extraction is performed by ultrasonic waves for 20 minutes, filtered and sterilized, and the filtrate is centrifuged to obtain a supernatant;

(6) Filtering the supernatant obtained in 5) sequentially using microfiltration membranes with pore sizes of 10 μm, 0.45 μm, and 0.2 μm, and collecting the filtrate to obtain the fermentation composition.

The drying method in step 1) is freeze drying;

In step 3), the wet ultrafine grinding is performed using a homogenizer.

The mass ratio of the peach resin, cactus and dendrobium is 3.5:2:2.

Table 1 shows the composition ratio of each fermentation composition of Examples 1 to 12 and Comparative Examples 1 to 3 (composition ratio of the cephalic acid: peach blossom: sedative composition is calculated by mass). Based on Table 1, fermentation compositions of each example and comparative example were prepared by the above method, and the effect of the obtained fermentation composition was tested.

Composition of raw materials (Chocolate: Peach Blossom: Calming composition, based on mass) Example 1 20:0.5:50 Example 2 20:2:50 Example 3 20:3.5:50 Example 4 20:5:50 Example 5 35:2:50 Example 6 50:2:50 Example 7 35:2:75 Example 8 35:2:100 Example 9 30:2:100 Example 10 40:2:100 Example 11 30:2:80 Example 12 30:2:70 Comparative Example 1 1:0:0 Comparative Example 2 0:1:0 Comparative Example 3 0:0:1

Unfermented group

The unfermented group in the following experimental examples of the present invention was prepared by the following process:

Here, the ratio of each composition was consistent with each Example and Comparative Example except that deionized water was used instead of the bacterial solution.

(1) Sorting and washing the eucalypts, peach blossoms, and soothing composition, drying them at low temperature, and then weighing them according to proportions;

(2) pulverizing peach resin, cactus, and dendrobium, then adding 20 times the total mass of deionized water of the eucalyptus, peach blossom, and soothing composition weighed in step 1), immersing them, and stirring for 2 hours;

(3) pulverizing the peach blossoms and peach blossoms, adding them to 2) and performing wet ultra-fine grinding to obtain a homogenate;

(4) adding 10% of the mass of deionized water to the homogenate obtained in 3) and stirring for 48 hours;

(5) ultrasonic extraction for 20 minutes, filtration and sterilization, and centrifugation of the filtrate to obtain a supernatant;

(6) filtering the supernatant obtained in 5) sequentially using microfiltration membranes with pore sizes of 10 μm, 0.45 μm, and 0.2 μm, collecting the filtrate, and obtaining an unfermented composition;

The low-temperature drying method in step 1) is freeze drying;

In step 3), the wet ultrafine grinding is performed using a homogenizer.

The mass ratio of the peach resin, cactus and dendrobium is 3.5:2:2.

Blank group

The blank group in the following experimental examples of the present invention was manufactured by the following process:

(1) taking a certain amount of deionized water;

(2) Adding 10% of the mass of the deionized water to deionized water and fermenting it with stirring at 35°C for 48 hours. The bacterial solution is lactic acid bacillus, and the bacterial content in the bacterial solution is ≥1.0×10 ¹¹ cfu/g. ;

(3) After fermentation is complete, ultrasonic extraction, filtration, and sterilization are performed for 20 minutes, and the filtrate is centrifuged to obtain a supernatant;

(4) Filtering the supernatant obtained in 3) sequentially using microfiltration membranes with pore sizes of 10 μm, 0.45 μm, and 0.2 μm, collecting the filtrate, and obtaining the blank group.

Experimental Example 1

Cytotoxicity test:

MTT assay is a widely used experimental method to detect cytotoxicity or cell proliferation by measuring the number of living cells. The detection principle is that succinate dehydrogenase in the mitochondria of living cells is activated by the water-soluble yellow salt MTT (3-(4). , 5-dimethylthiazole-2)-2, 5-diphenyltetrazolium bromide) is reduced to water-insoluble blue formazan and accumulated within cells. Dead cells do not have this function. The produced crystalline formazan is generally dissolved by adding it to DMSO (dimethyl sulfoxide) and then the absorbance is measured. The specific experimental method is to inoculate 100 μl each of DMEM medium containing 10% bovine serum and keratinocytes (HaCaT) at a density of 1Х10 ⁴ cells/well in a 96-well plate, culture for 24 hours, and then incubate in serum-free medium. is to be replaced with . Compositions of different concentrations were added to the serum-free medium, treated, and cultured for 24 hours. After removing the medium, it was treated with 20 μl of MTT solution and reacted at 37°C for 2 hours. 200 μl of isopropanol was added to the cells from which the MTT solution was removed and shaken lightly for 30 minutes to completely dissolve the crystalline formazan. Absorbance was measured at 570 nm, and cell viability was calculated according to the formula below.

The blank group was tested without adding fermentation composition. Table 2 shows the cytotoxicity results of the fermentation compositions obtained in Examples 1 to 12 and Comparative Examples 1 to 3.

Group classification Experimental concentration (μg/ml) Cell viability (%) Blank group 100 Example 1 50 96.01 100 96.27 200 95.33 Example 2 50 96.64 100 95.87 200 96.34 Example 3 50 97.16 100 97.31 200 95.69 Example 4 50 96.61 100 95.14 200 94.89 Example 5 50 97.07 100 96.78 200 96.32 Example 6 50 97.77 100 97.36 200 96.52 Example 7 50 97.18 100 96.64 200 96.15 Example 8 50 97.02 100 96.48 200 96.43 Example 9 50 97.12 100 96.87 200 96.26 Example 10 50 96.98 100 96.38 200 95.79 Example 11 50 97.58 100 96.85 200 96.88 Example 11 50 96.98 100 96.29 200 95.88 Comparative Example 1 50 96.22 100 96.04 200 95.65 Comparative Example 2 50 96.56 100 95.32 200 95.01 Comparative Example 3 50 96.32 100 95.69 200 95.34

Experimental Example 2

Antioxidant evaluation test:

DPPH, also called 1,1-diphenyl-2-trinitrophenylhydrazine, is a very stable nitrogen-centered free radical. Its anhydrous ethanol solution appears purple, and when a free radical scavenger is added, the number of free radicals decreases, causing the absorbance to decrease. The lower the value, the lighter the color of the solution, so the free radical scavenging ability can be evaluated.

Take 40 μl of the obtained composition with a pipette, and add 160 μl of DPPH ethanol solution (200 μg/ml). After mixing uniformly, the reaction was performed for 30 minutes at room temperature under light-shielding conditions, and the absorbance A was measured again at 517 nm. Three parallel experiments were set up for each group to calculate the average value. The calculation method was as follows:

Erasing rate % =[1-（A ₁ -A ₂ ）/A ₀ ]Х100

In the formula, A ₀ is the absorbance of the DPPH solution without the addition of each fermentation composition;

A ₁ is the absorbance after reaction of the DPPH solution after adding each fermentation composition;

A ₂ is the absorbance before reaction after mixing the DPPH solution and each fermentation composition.

Table 3 shows the results of the antioxidant evaluation test of each fermented composition in Examples 1 to 12 and Comparative Examples 1 to 3.

Composition of raw materials (Chocolate: Peach Blossom: Calming composition, based on mass) Average extinction rate (%) Example 1 20:0.5:50 61.96 Example 2 20:2:50 63.34 Example 3 20:3.5:50 63.78 Example 4 20:5:50 63.86 Example 5 35:2:50 65.54 Example 6 50:2:50 66.11 Example 7 35:2:75 72.68 Example 8 35:2:100 68.43 Example 9 30:2:100 69.22 Example 10 40:2:100 66.86 Example 11 30:2:80 72.13 Example 12 30:2:70 69.76 Comparative Example 1 1:0:0 51.97 Comparative Example 2 0:1:0 56.12 Comparative Example 3 0:0:1 52.26

From the table above, it can be seen that:

The DPPH removal rates of the fermentation compositions obtained in Examples 1 to 12 are all higher than those in Comparative Examples 1 to 3, which explains that a better DPPH removal effect can be obtained by combining the three raw materials;

The DPPH removal rate has a constant dose-response relationship for all three raw materials, and when the mass fraction increases to a certain range, the upward trend slows down. As a result of comparing Examples 1, 2, 3, and 4, it was found that the difference in the removal rate of DPPH when the mass fraction of peach blossom was 2 and 5 was not significant; As a result of comparing Examples 2, 5, and 6, it was found that the difference in DPPH removal rate was not significant when the mass fraction of cephalic acid was 35 and 50; As a result of comparing Examples 5, 7, 8 and 9, 11, and 12, it was found that the removal rate was the best when the mass fraction of the soothing composition was 75 and 80. Therefore, considering the DPPH removal ability, economic effect and advantages, Example 7 (35:2:75) and Example 11 (30:2:80) are preferred.

Summarizing the above, when the three raw materials are combined in different ratios, the corresponding antioxidant ability is different, and the mass ratio of the cephalic acid: peach blossom: soothing composition is (30-40): (1-3): (60-). 90), and it is more preferable that the mass ratio of the cephalic acid:peach blossom:sedative composition is 35:2:75 and 30:2:80.

Experimental Example 3

MMP-1 gene expression test:

MMP-1 (Matrix metalloproteinase-1) is the main enzyme responsible for collagen degradation, and can be upregulated after sunlight or infrared irradiation. Since collagen degradation is an important indicator of photoaging, expression of MMP-1 has an anti-photoaging effect. It is used as an indicator for evaluation.

Using the ELISA method, negative group (no sample added, treated with IR irradiation), blank group (fermented product without added composition, treated with IR irradiation), Examples 7-1, 7-2 (0.1% and 0.3% sample added, treated with IR irradiation), unfermented Example 7-1, unfermented Example 7-2 (added 0.1% and 0.3% of unfermented composition, treated with IR irradiation), Examples 11-1, 11-2 (0.1% and 0.3% of sample added, treated with IR irradiation), unfermented Example 11-1, unfermented Example 11-2 (0.1% and 0.3% of fermented composition added, treated with IR irradiation), Comparative Example 1 -1, 1-2 (added 0.1% and 0.3% of sample, treated with IR irradiation), Comparative Example 2-1, 2-2 (added 0.1% and 0.3% of sample, treated with IR irradiation), Comparative Example 3 Human MMP-1 was detected from -1 and 3-2 (0.1% and 0.3% of sample added, treated with IR irradiation). 100 μl _of the sample to be tested (10 Each cell plate is set as a negative control (cells + medium), and each concentration group has three replicate wells. Cell culture supernatants were collected, and the collagen degradation effect was evaluated using an ELISA detection kit.

Calculation formula: MMP-1 inhibition rate % = (OD ₄₅₀ negative group - OD ₄₅₀ test target sample group) / OD ₄₅₀ negative group × 100.

In MMP-1 expression, as shown in Figure 1, the blank group has no significant effect on MMP-1 expression, and Example 7 (0.1%, 0.3%) and Example 11 (0.1%, 0.3%) are compared. Compared to Examples 1 to 3, the expression of MMP-1 could be significantly lowered. Additionally, the fermented composition shows a more significant effect in reducing MMP-1 compared to the unfermented composition. It was demonstrated that the effect of improving skin photoaging of this fermented composition is superior to that of a single ingredient and that the effect of improving photoaging after fermentation is more excellent.

Experimental Example 4

In vitro antiglycation activity test:

The anti-glycation ability of the fermentation composition was evaluated by creating a bovine serum albumin (BSA)-Glucose model to simulate the Maillard reaction system. The reaction mixture contained bovine serum albumin (BSA), glucose, phosphate buffer solution (PBS), and each fermented and unfermented composition sample (Assample), and was cultured for 28 days at 37°C under sterile conditions. Among these, aminoguanidine (aminoguanidine; A sample that does not contain AG) and fermentation composition is a negative control (ANC), and a sample in which AG is added instead of polysaccharide is a positive control (Assample). The fluorescence intensity of the advanced glycation product was measured using a detection method using fluorescence spectroscopy, and the excitation wavelength was set to 370 nm and the emission wavelength was set to 440 nm. The inhibition rate (R) of different samples against the production of fluorescent AGEs was calculated using the formula:

R（%）=（ANC－Asample）/ ANCХ100%

Among them, ANC is the absorbance of negative control; Asample is the absorbance of the fermentation composition or AG group.

As shown in Figure 2, the inhibition rates for the production of AGEs in Examples 7 and 11 are all higher than those of Comparative Examples 1, 2, and 3, resulting in a synergistic effect when the fermentation composition is mixed in an optimal combination. It has been proven that it can improve the anti-glycation effect.

Experimental Example 5

Safety skin patch testing:

Drop 20 μl of test liquid into the patch tester, and set the control well as a blank control (pure water); Attach the patch tester containing the test substance to the front curved side of the subject's arm, apply it evenly to the skin by lightly pressing it with the palm, and keep it for 24 hours; After removing the patch tester, observe skin irritation and allergenicity according to Table 4 after 30 minutes, 24 hours, and 48 hours, and record the observation results. The grading criteria for skin reaction in the skin occlusive patch test are shown in Table 4.

evaluation grade skin reaction 0 voice response One Suspicious reaction: Only slight erythema appears. 2 Mildly benign reaction (erythematous reaction): erythema, infiltrates, dropsy, and papules may occur. 3 Strongly positive reaction (herpetic reaction): erythema, infiltrates, dropsy, papules, herpes, possibility of reaction exceeding test area. 4 Very strong benign reaction (confluent herpetic reaction): marked erythema, severe infiltrates, hydroceles, confluent papules, reaction exceeds the test area

Experimental results: Human skin patch tests were conducted in Examples 7 and 11, and the results are shown in Table 5. As a result of the human skin patch test, none of the 30 people showed any side effects on the skin.

Test items and times water Example 7 Example 11 0.5 hours 24 hours 48 hours 0.5 hours 24 hours 48 hours 0.5 hours 24 hours 0.5 hours Number of people with various skin reactions in patch tests 0 20 20 20 20 20 20 20 20 20 One 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 3 0 0 0 0 0 0 0 0 0 4 0 0 0 0 0 0 0 0 0

Experimental Example 6

1) Cooling effect test:

30 female volunteers between the ages of 25 and 45 were selected, Example 7 was applied to 10 volunteers, Example 11 was applied to 10 volunteers, and the blank group (placebo) was applied to the remaining 10 volunteers. Apply a unit dose of 2 mg/cm ² to the entire face, use it every morning and evening, use it for 2 consecutive weeks, and use a skin surface temperature-thermal imaging camera (FLIR T450 sc, USA) on the first day and after 2 weeks. The cooling effect of the fermentation composition was evaluated.

The experimental results are as shown in Table 6, and as a result of comparing the blank group with the emulsion containing Example 7 and Example 11, it was found that the fermentation composition of the present invention had a very excellent cooling effect.

Group classification Before(℃) 2 weeks (℃) 2 weeks after IR (℃) Blank group 34.1 ± 0.48 33.9 ± 0.54 36.2 ± 0.20 Example 7 33.4 ± 0.36 32.2 ± 0.28 35.2 ± 0.22 Example 11 33.6 ± 0.38 32.1 ± 0.24 35.6 ± 0.32

2) Skin elasticity test:

After continuous use for 14 days, the elastic recovery ratio (R2) (A.U.) before use, 2 weeks, and 4 weeks after use was tested using the device. R2: elasticity recovery without negative pressure and maximum elongation with negative pressure. In terms of the ratio of the amount, the closer the ratio is to 1, the better the skin elasticity. The results are shown below respectively.

Experimental results: The results are shown in Figure 3, and in Examples 7 and 11, skin elasticity was significantly improved compared to the blank group, and was improved by 10.4% and 10.5% compared to skin elasticity before use.

3) Skin barrier recovery test:

Figure 4 is a graph of percent reduction in transepidermal water loss (Twel) values after using different fermentation compositions. From Figure 4, it can be seen that Examples 7 and 11 can improve the transepidermal moisture loss of the skin, restore the damaged skin barrier, and improve the anti-allergy and anti-irritant abilities of the skin.

4) Skin texture test:

Figure 5 is a view showing skin texture before and after using different fermentation compositions. From Figure 5, it can be seen that Examples 7 and 11 significantly improved skin texture.

In the specification of the present application, information that is not described in detail corresponds to common knowledge of those skilled in the art.

As mentioned throughout the specification and claims, “include” is an open term and should be interpreted as “including but not limited thereto.” “Approximately” refers to existing within an allowable error range, and those skilled in the art can solve the above technical problems and basically achieve the above technical effects within a certain error range.

It is also important to note that terms such as "comprise" or any other variation thereof are intended to be non-exclusive, meaning that a product or system containing a set of elements may include those elements as well as other elements not explicitly listed. This means that it includes elements or elements unique to this product or system. Unless further limitation exists, elements defined by the expression “including…” do not exclude the presence of other identical elements in a product or system containing said elements.

Although some preferred embodiments of the present application have been shown and described through the above description, as described above, the present application is not limited to the form disclosed in the text, and should not be considered to exclude other embodiments, and various other combinations and modifications. Alternatively, it can be applied to the environment, and can be improved through the above teachings or technology or knowledge in related fields within the scope of the invention concept described in the text. Improvements and changes made by those skilled in the art do not depart from the spirit and scope of the present application, and must all fall within the scope of protection of the claims of the present application.

Claims (10)

A fermentation composition having anti-photoaging, soothing and cooling effects, obtained by grinding, fermenting and extracting eucalyptus, peach blossom and soothing composition having a mass ratio of 20 to 50:0.5 to 5:50 to 100. Obtained;
The soothing composition is a combination of peach resin, cactus and dendrobium; A fermentation composition, characterized in that the mass ratio of the peach resin, cactus and dendrobium is 3:1:1 or 3.5:2:2. The fermented composition according to claim 1, wherein the mass ratio of the Eucommia:peach blossom:sedative composition is 30 to 40:1 to 3:60 to 90. The fermented composition according to claim 1, wherein the mass ratio of the Eucommia:peach blossom:sedative composition is 35:2:75 or 30:2:80. The method of any one of claims 1 to 3, wherein the manufacturing method specifically
1) Sorting and washing the eucalyptus, peach blossom, and soothing composition, drying at low temperature, and then weighing them according to ratio;
2) pulverizing peach resin, cactus, and dendrobium, adding them to deionized water, immersing them, and stirring for 2 hours;
3) pulverizing the peach blossoms and peach blossoms, adding them to 2) and performing wet ultra-fine grinding to obtain a homogenate;
4) inoculating the homogenate obtained in 3) with the bacterial solution and fermenting it while stirring at a uniform speed;
5) After fermentation is complete, ultrasonic extraction, filtration and sterilization are performed, and the filtrate is centrifuged to obtain a supernatant; and
6) Filtering the supernatant obtained in 5) sequentially using microfiltration membranes having different pore sizes, collecting the filtrate, and obtaining the fermentation composition. According to clause 4,
The low-temperature drying conditions in step 1) are drying below 50°C; The drying method includes one or a combination of oven drying, freeze drying, and vacuum drying;
The volume of deionized water in step 2) is 5 to 20 times the total mass of eucalyptus, peach blossom and soothing composition;
In step 3), the wet ultra-fine grinding is performed using a colloid mill or homogenizer. According to clause 4,
In step 4), the inoculating bacterial solution contains one or a combination of two of lactic acid bacilli, lactic acid bacteria, and yeast;
In step 4), the inoculum inoculation amount is 5% to 10% of the mass of the homogenate, the bacterial content is ≥1.0Х10 ¹¹ cfu/g, the fermentation temperature is 28 to 35°C, and the fermentation time is 12 to 48 hours. Characterized by fermentation composition. According to clause 4,
Fermentation composition, characterized in that the pore size range of the microfiltration membrane in step 6) is 0.1 to 10 μm. A method for producing a fermentation composition, characterized in that it comprises the following steps:
1) Sorting and washing the eucalyptus, peach blossom, and soothing composition, drying at low temperature, and then weighing them according to ratio;
2) the soothing composition is a combination of peach resin, cactus and dendrobium; Grinding the peach resin, the cactus, and the dendrobium, adding them to deionized water, immersing them, and stirring for 2 hours;
3) pulverizing the peach blossoms and peach blossoms, adding them to 2) and performing wet ultra-fine grinding to obtain a homogenate;
4) inoculating the homogenate obtained in 3) with the bacterial solution and fermenting it while stirring at a uniform speed;
5) After fermentation is complete, ultrasonic extraction, filtration and sterilization are performed, and the filtrate is centrifuged to obtain a supernatant; and
6) filtering the supernatant obtained in 5) using microfiltration membranes having different pore sizes sequentially and collecting the filtrate to obtain the fermentation composition;
The low-temperature drying conditions in step 1) are drying below 50°C; The drying method includes one or a combination of oven drying, freeze drying, and vacuum drying;
The volume of deionized water in step 2) is 5 to 20 times the total weight of eucalyptus, peach blossom, and soothing composition;
In step 3), the wet ultrafine grinding is performed using a colloid mill or homogenizer;
In step 4), the inoculating bacterial solution contains one or a combination of two of lactic acid bacilli, lactic acid bacteria, and yeast; The inoculant inoculum amount is 5% to 10% of the homogenate mass, the bacterial content is ≥1.0Х10 ¹¹ cfu/g, the fermentation temperature is 28 to 35°C, and the fermentation time is 12 to 48 hours;
The manufacturing method, characterized in that the pore size range of the microfiltration membrane in step 6) is 0.1 to 10 μm. 9. The method of claim 8, wherein the mass ratio of the soothing composition composed of eucalyptus, peach blossom, and peach resin, cactus and dendrobium is 30 to 40:1 to 3:60 to 90;
A manufacturing method characterized in that the mass ratio of the peach resin, cactus and dendrobium is 3:1:1 or 3.5:2:2. Containing the fermentation composition of claim 1; Cosmetics, characterized in that they are any one of liquid, emulsion, spray, cream, mask pack, and liquid foundation.

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