CN112353734A - Fermentation method of centella fermented product and application of centella fermented product in repair-efficacy cosmetics - Google Patents

Abstract

The invention belongs to the field of separation of natural plant active ingredients and cosmetic additives, and particularly relates to application of a centella fermented product in preparation of a product with a repairing effect. The invention provides application of a centella fermented product in repairing efficacy, wherein the centella fermented product has good in-vivo cell repairing activity. The invention also provides a fermentation method of the asiatic pennywort herb fermentation product, so that the content of the asiatic pennywort herb polysaccharide serving as the effective component in the fermentation liquid is up to 0.54 percent.

Description

Fermentation method of centella fermented product and application of centella fermented product in repair-efficacy cosmetics

Technical Field

The invention belongs to the field of separation of natural plant active ingredients and cosmetic additives, and particularly relates to a repair-effect cosmetic containing a centella fermented product and a preparation method thereof.

Background

Centella asiatica (L.) Urb, as a traditional Chinese medicine, has bitter taste and cold nature, enters liver, spleen and kidney meridians, has the effects of clearing heat, promoting diuresis, removing toxic substances and relieving swelling, and is derived from dried whole plant of Centella asiatica of Centella in Umbelliferae, also called as Callicarpa, Eleocharis Turerosa, copper coin grass, denseflower, Breviscapine, Desmodium styracifolium and the like. At present, the fertilizer is mainly distributed in places such as Anhui, Zhejiang, Hunan, Hubei, Jiangxi, Guangdong and the like in China, the growing environment is a multi-tide wetland or a low grass cluster, the fertilizer is favorable for dampness and yin resistance, and the fertilizer can be harvested all the year round. Centella asiatica was originally recorded in Shen nong Ben Cao Jing, and has been used for oral administration and external application, and has been used for over 2000 years, and the main effective components comprise asiaticoside, madecassoside, asiatic acid, madecassic acid and the like.

The natural product extract is prepared by a microbial fermentation process, so that inherent toxicity of some natural products can be effectively reduced, the cell wall structure of medicinal materials can be subjected to enzymolysis in the fermentation process, effective components of the natural products are released more fully, and the utilization rate of the natural products is improved. In the fermentation process, microorganisms also produce secondary metabolites, wherein the secondary metabolites comprise a plurality of biological enzymes, and can effectively decompose biological macromolecules such as proteins, polysaccharides and the like into substances with smaller molecular weight, and the biological macromolecules take the proteins as an example, the proteins can be hydrolyzed by protease into biological active peptides with the same sequence as functional proteins, small specificity and different molecular weights. Research shows that the fermented extract can raise the matter content of the effective component and thus raise various pharmacological activities obviously. In addition, the microbial fermentation has the characteristics of low cost, difficult sensitization and the like, and is a hotspot of current research and development.

The skin, the largest organ of the human body, is exposed to various injuries outside the body, wherein acute injury and chronic injury caused by ultraviolet radiation are the most serious, when the skin is excessively exposed to UVA (400-315 nm) and UVB (315-280 nm) from sunlight, acute dermatitis is caused, the symptoms of the acute dermatitis comprise pruritus, pain, erythema, edema, desquamation and the like, and the skin can even become cancerous after being exposed to ultraviolet radiation for a long time. Research shows that solar dermatitis is always acute inflammation caused by exposure of normal skin, and has great damage to the skin, but the main effective components of a type of after-sun repair products sold on the market at present are cell activating factors, so that the after-sun damage cannot be reduced and repaired, and the repair function for the solar acute dermatitis and the secondary photosensitive dermatitis is lacked.

Disclosure of Invention

Aiming at a series of muscle bottom problems commonly existing at present, the invention provides a preparation method of a centella fermented product and application of the centella fermented product in cosmetics with repairing effects. The centella fermented product provided by the invention has a remarkable repairing effect. The invention also provides a preparation method of the asiatic centella fermented product, and the preparation method has the advantages of simple fermentation process, capability of reducing fermentation cost and easiness in expanding production. The centella asiatica fermentation liquor is rich in various plant secondary metabolites taking polysaccharide as the main component, and is converted into an effective component with smaller molecular weight under the condition of microbial fermentation, so that the effective component is easier to be absorbed and utilized by a human body, and simultaneously, the centella asiatica fermentation product obtained by the fermentation of the method is a high-safety repair-effect cosmetic raw material.

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

the invention provides application of a centella fermented product in preparation of a product with a repairing effect.

The invention also provides application of the asiatic centella fermentation product in preparation of cosmetics with repairing efficacy.

The centella asiatica fermentation product provided by the invention has the effect of relieving the cytotoxicity of common cosmetic preservatives (such as phenoxyethanol), and can greatly improve the safety of cosmetics with a repairing effect.

The invention provides a centella fermented product, wherein the content of centella polysaccharide in the centella fermented product is 0.54%.

The fermentation method provided by the invention has the advantages of simple fermentation process, low fermentation cost and easy expanded production; can improve the content of the effective component asiatic pennywort herb polysaccharide in the fermentation product and can greatly improve the utilization degree of the asiatic pennywort herbs. The specific fermentation process is as follows: accurately weighing asiatic pennywort herb leaves, crushing and sieving to obtain asiatic pennywort herb leaf powder, wherein the particle size of the powder is preferably 120 meshes, and then the powder is prepared by the following steps of: (10-12) (g/mL) adding sterile deionized water, and adjusting the pH value to 6.0-6.5; in the present invention, the feed-to-liquid ratio of the fermentation is more preferably 1: 10, the deionized water sterilization condition is further preferably at 121 deg.C and 0.1MPa for 20 min.

After the mixture of the centella asiatica night-film powder and the deionized water is obtained, inoculating Saccharomyces cerevisiae seed liquid (Saccharomyces cerevisiae) according to the feeding proportion of 1 per mill, fully stirring to uniformly mix the powder, setting the temperature of a fermentation tank to be 30 ℃, and fermenting for 60-72 hours to start fermentation; in the invention, the preparation method of the saccharomyces cerevisiae seed liquid is preferably as follows: sterilizing a wort culture medium at 121 ℃ and 0.1MPa, inoculating saccharomyces cerevisiae, setting the culture temperature at 28 ℃ and the rotating speed of a shaking table at 120 r/min, and culturing for 24 hours; the cell concentration is set to 108CFU/mL, and the fermentation time is more preferably 72 hours.

After fermentation, centrifuging at 8000-10000 rpm/min, taking supernate to break the wall under the condition of 1150-1200 Bar, cooling, filtering through a 0.45 mu m microporous filter membrane to obtain fermentation product filter residue and filtrate, and taking the fermentation product filtrate as a centella fermented product; in the invention, the centrifugation condition is further optimized to 8000rpm/min, and the wall breaking condition is further optimized to 1200 Bar.

According to the invention, through optimizing each step of the fermentation engineering, the efficiency and effect of fermentation can be effectively improved by mutually matching a plurality of optimization steps. The content of centella asiatica polysaccharide in the centella asiatica fermentation product obtained by fermentation by the fermentation method is 0.54%.

In the present invention, the source of the above-mentioned fermentation material and other products is not particularly limited, and conventional commercially available products known to those skilled in the art may be used.

The invention also provides a repair-effect cosmetic containing the asiatic pennywort herb fermentation product, and the repair-effect product comprises the asiatic pennywort herb fermentation product obtained by the fermentation method. The repair effect product provided by the invention has an excellent repair effect, can obviously reduce the loss of the zebra fish caused by ultraviolet radiation, and has a good repair effect on the damage caused by the ultraviolet radiation.

The invention also provides a repair-effect cosmetic based on the asiatic centella fermented product or the asiatic centella fermented product obtained by the fermentation method, which comprises the following components in parts by mass: cetearyl/sorbitan olive oleate 2.00%, pentaerythritol distearate 1.00%, ethylhexyl palmitate 5.00%, caprylic/capric triglyceride 3.00%, tocopherol 0.5%, C18-C21 alkane 5.00%, xanthan gum 0.25%, allantoin 0.30%, glycerol 3.00%, centella asiatica fermentation product 0.50%, phenoxyethanol/ethylhexyl glycerol 0.50%, essence 0.30%, and the balance of deionized water.

In the present invention, the above-mentioned cosmetic preparation step preferably comprises the steps of:

1) preparing an oil phase: firstly, accurately weighing cetearyl olive oleate/sorbitan olive oleate, pentaerythritol distearate, ethylhexyl palmitate, caprylic/capric triglyceride, tocopherol and C18-C21 alkane, and heating to 80 ℃ to completely melt;

2) preparing an aqueous phase: accurately weighing xanthan gum, allantoin and glycerol, adding into deionized water required by the system, heating to 80 deg.C, stirring and dissolving to transparent;

3) adding the water phase into the oil phase, and homogenizing at a speed of 10000rpm/min for 30 s-1 min;

4) stirring the system, cooling to room temperature, sequentially adding herba Centellae fermentation product, phenoxyethanol/ethylhexyl glycerol and essence, and continuously stirring; cooling, aseptic standing for 24 hr, and packaging to obtain the final product.

The sources of the above-mentioned adjuvants in the present invention are not particularly limited, and conventional commercially available products known to those skilled in the art may be used.

In order to further illustrate the present invention, the use of the fermented products of centella asiatica in the preparation of products with restorative effect is described in detail below with reference to the accompanying drawings and examples, which should not be construed as limiting the scope of the present invention.

Drawings

FIG. 1 shows a fermentation process of centella asiatica.

Detailed Description

Example 1

Safety test of centella asiatica fermentation product

1 method of experiment

1.1 Collection of Fish for experiment and fertilized ovum thereof

And feeding the sexually mature zebra fish in the zebra fish culture unit in separate jars. Water temperature: 26 +/-2 ℃; pH 7.2; conductivity: 520 mus/cm; light/dark cycle: 14 h:10 h. The zebra fish were paired at a ratio of 1:2 in a male-female ratio the day before the exposure experiment was started, and naturally mated and spawned under light stimulation.

1.2 Exposure experiments

1) Healthy zebrafish embryos which develop to 0-3 hpf (hour post-fertilization) are selected and placed in 6-well cell culture plates, 20 fish/well, and 5mL of experimental solution is added into each well. Zebra fish culture water was used as a blank control group.

2) The number of hatchings, malformations and deaths of fish embryos was observed and counted under a stereomicroscope at 24, 48 and 72 hpf, respectively. Wherein the malformed configuration comprises: head deformity, pericardial edema, yolk sac edema, blood blockage, spinal curvature, tail curvature, developmental retardation, etc.; the death state is judged as follows: the yolk sac coagulates and the heartbeat stops.

2 results of the experiment

The effect of centella asiatica fermentation products on fish embryo mortality, hatchability and teratogenicity is shown in tables 1, 2 and 3.

TABLE 1 Effect of centella asiatica fermentation products on mortality (%) of fish embryos

(comparison with blank control group:. P < 0.05, P < 0.01, P < 0.001)

TABLE 2 Effect of centella asiatica fermentation products on the hatchability (%) of fish embryos

(comparison with blank control group:. P < 0.05, P < 0.01, P < 0.001)

TABLE 3 Effect of centella asiatica fermentation products on the distortion rate (%) of fish embryos

(comparison with blank control group:. P < 0.05, P < 0.01, P < 0.001)

Note: mortality% = number of dead embryos/N100%; hatchability% = number of hatched embryos/number of surviving embryos 100%; percent teratogenicity rate = (number of dead embryos + number of malformed embryos)/N100%

As can be seen from the analysis of tables 1, 2 and 3, the asiatic pennywort herb fermentation product has no significant difference in three aspects of lethal dose, hatchability and aberration rate when the concentration of the asiatic pennywort herb fermentation product is up to 8%, and the conclusion can be drawn that the 0.2% -1% asiatic pennywort herb fermentation product used in the formula is absolutely safe and has no toxic or side effect.

Example 2

Experiment for relieving negative effects of traditional preservative by centella asiatica fermentation product

1 method of experiment

In general, a preservative is added to a cosmetic formulation for preservation. Preservatives present a certain toxicity. The common preservative phenoxyethanol is selected for testing the preservative effect of the asiatic pennywort herb fermentation product in the experiment. Referring to the experimental method of example 4, a blank control group, a group of 1% phenoxyethanol and a group of centella asiatica fermentation products (hereinafter referred to as samples) added with 1% phenoxyethanol were set, and the influence of the blank control group, the group of 1% phenoxyethanol and the group of centella asiatica fermentation products on the death rate and the teratogenicity rate of fish embryos was investigated.

2 results of the experiment

2.1 Effect of samples on safety of Fish embryos

(1) The effect of 1% phenoxyethanol on mortality and teratogenicity of fish embryos is shown in tables 4 and 5.

TABLE 41 Effect of phenoxyethanol on mortality (%) of fish embryos

(comparison with blank control group:. P < 0.05, P < 0.01, P < 0.001)

TABLE 51% Effect of phenoxyethanol on Fish embryo distortion (%)

(comparison with blank control group:. P < 0.05, P < 0.01, P < 0.001)

2.2 the effect of the samples on mortality and teratogenicity of fish embryos is shown in tables 6 and 7.

TABLE 6 Effect of samples on mortality (%) of Fish embryos

(comparison with blank control group:. P < 0.05, P < 0.01, P < 0.001)

TABLE 7 influence of samples on the distortion rate (%) of fish embryos

(comparison with blank control group:. P < 0.05, P < 0.01, P < 0.001)

Mortality% = number of dead embryos/N100%; percent teratogenicity rate = (number of dead embryos + number of malformed embryos)/N100%

Analysis of tables 4 and 6 shows that the synergistic effect of the asiatic centella fermentation product and the phenoxyethanol has no influence on the lethality rate of the phenoxyethanol, but comparison of tables 5 and 7 shows that the asiatic centella fermentation product can weaken the teratogenic effect of the phenoxyethanol, which is a traditional preservative, to a certain extent.

Example 3

Test of repairing efficacy of centella asiatica fermentation product

1 post-basking repair Activity study experiment

Healthy AB line zebrafish embryos of 72 hpf after fertilized egg selection are used for experimental study, and are placed on 6-well cell culture plates with 1 cell per well, and 5ml of experimental solution is added into each well. A blank control group (zebra fish culture water) model control group is additionally arranged in the experiment, and each group is provided with 3 multiple holes;

(2) irradiating with ultraviolet analyzer (305 nm wavelength) for 30s every half an hour within 2.5 hr, absorbing the sample liquid, changing into culture water, and incubating in incubator;

(3) observing the damage conditions of the ventral fin, the tail fin and the dorsal fin of the zebra fish under an inverted microscope on days 1 and 2 after ultraviolet radiation, and photographing and recording;

(4) and analyzing the damage degree and the repair degree of the tail fin of the zebra fish juvenile fish by using software and calculating.

2 post-basking repair Activity test results

The damage of centella asiatica fermentation product to zebra fish after sun exposure and the repairing effect thereof are shown in table 8. As can be seen from Table 8, the zebra fish embryos of the blank control group have large-area damage after receiving ultraviolet radiation, and have poor self-repairing effect after 48 hours; the damage to the zebra fish embryos of the asiatic pennywort herb fermentation product treatment groups with different concentrations is obviously reduced, the repair rate is also obviously improved after 48 hours, wherein the effect is enhanced along with the increase of the concentration of the asiatic pennywort herb fermentation product, namely when the concentration of the asiatic pennywort herb fermentation product is more than or equal to 0.2%, the zebra fish embryos have the ultraviolet radiation resistance and after-sun repair effects and are positively correlated with the concentration.

TABLE 8 influence of fermentation products of centella asiatica on post-sun damage and repair

(Note: with statistical differences, P < 0.01).

Claims (9)

1. Application of herba Centellae fermentation product in preparing product with repairing effect is provided.

2. Application of herba Centellae fermentation product in preparing cosmetic with repairing effect is provided.

3. A centella asiatica fermented product, wherein the content of centella asiatica polysaccharide in the centella asiatica fermented product is 0.54%.

4. A process for the fermentation of a product of centella asiatica fermentation in the use according to claim 1 or 2 or of a product of centella asiatica fermentation according to claim 3, comprising the following steps: the centella asiatica fermentation product is prepared by the following microbial fermentation method, and the specific steps are as follows: (1) accurately weighing herba Centellae leaves, pulverizing, and sieving with 120 mesh sieve; (2) according to the following steps: (10-12) (g/mL) adding sterilized deionized water, and adjusting the pH to 6.0-6.5;

(3) inoculating Saccharomyces cerevisiae seed liquid (Saccharomyces cerevisiae) according to the feeding proportion of 1 per mill, uniformly stirring, setting the temperature of a fermentation tank to be 30 ℃, and fermenting for 60-72 hours to start fermentation; (4) centrifuging at 8000-10000 rpm/min after fermentation is finished, taking supernate to finish wall breaking under the condition of 1150-1200 Bar, cooling, and filtering and removing slag through a 0.45 mu m microporous filter membrane to obtain fermentation product filtrate.

5. The sterilized deionized water according to claim 2, wherein the sterilization is performed at 121 ℃ and 0.1MPa for 20 min.

6. The preparation method of the saccharomyces cerevisiae seed liquid culture medium according to the claim (3), characterized in that after the wort culture medium is sterilized at 121 ℃ and 0.1MPa for 20min, saccharomyces cerevisiae is inoculated, the culture temperature is set at 28 ℃, the rotation speed of a shaker is set at 120 r/min, and the culture is carried out for 24 h; the cell concentration was adjusted to 108CFU/mL, respectively.

7. A restoration effect product containing a fermentation product of centella asiatica, wherein the restoration effect product comprises the fermentation product of centella asiatica fermented by the method of claim 3 and auxiliary materials.

8. A repair-efficacy cosmetic containing a centella asiatica fermentation product based on the centella asiatica fermentation product of claim 3, comprising the following components in parts by mass: cetearyl/sorbitan olive oleate 2.00%, pentaerythritol distearate 1.00%, ethylhexyl palmitate 5.00%, caprylic/capric triglyceride 3.00%, tocopherol 0.5%, C18-C21 alkane 5.00%, xanthan gum 0.25%, allantoin 0.30%, glycerol 3.00%, centella asiatica fermentation product 0.50%, phenoxyethanol/ethylhexyl glycerol 0.50%, essence 0.30%, and the balance of deionized water.

9. A method for preparing a cosmetic with restorative effect based on the fermentation product of centella asiatica of claim 3, comprising the steps of:

1) preparing an oil phase: firstly, accurately weighing cetearyl olive oleate/sorbitan olive oleate, pentaerythritol distearate, ethylhexyl palmitate, caprylic/capric triglyceride, tocopherol and C18-C21 alkane, and heating to 80 ℃ to completely melt;

2) preparing an aqueous phase: accurately weighing xanthan gum, allantoin and glycerol, adding into deionized water required by the system, heating to 80 deg.C, stirring and dissolving to transparent;

3) adding the water phase into the oil phase, and homogenizing at a speed of 10000rpm/min for 30 s-1 min;

4) stirring the system, cooling to room temperature, sequentially adding herba Centellae fermentation product, phenoxyethanol/ethylhexyl glycerol and essence, and continuously stirring; cooling, aseptic standing for 24 hr, and packaging to obtain the final product.

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