CN116549345A

CN116549345A - Oil-control and soothing plant fermentation product and preparation method and application thereof

Info

Publication number: CN116549345A
Application number: CN202310552766.XA
Authority: CN
Inventors: 何敬愉; 林琳; 潘丹阳; 徐崇; 张得香
Original assignee: Guangzhou Huanya Cosmetic Science and Technology Co Ltd
Current assignee: Guangzhou Huanya Cosmetic Science and Technology Co Ltd
Priority date: 2023-05-17
Filing date: 2023-05-17
Publication date: 2023-08-08

Abstract

The invention belongs to the technical field of microbial fermentation, and discloses an oil-control and soothing plant fermentation product, and a preparation method and application thereof. The preparation raw materials of the plant ferment comprise: plant material, cow milk, glycerol and fermentation strains; the plant material comprises at least one of cacumen Platycladi, folium Platycladi, and folium Selaginellae; the fermentation strain comprises at least one of lactobacillus plantarum, lactobacillus lactis and bifidobacterium longum. According to the invention, the high-pressure homogenization and directional microorganism fermentation technology are organically combined, cow milk and glycerol are assisted in the high-pressure homogenization process, so that plant tissue cells can be fully crushed, active ingredients are fully released and dissolved, the utilization efficiency of raw materials is improved, the composite stock solution obtained by the high-pressure homogenization is used as a subsequent fermentation substrate, and the prepared plant fermentation product has a remarkable oil control and relief effect through the bioconversion capability of microorganisms.

Description

Oil-control and soothing plant fermentation product and preparation method and application thereof

Technical Field

The invention belongs to the technical field of microbial fermentation, and particularly relates to an oil-control and soothing plant fermentation product, and a preparation method and application thereof.

Background

In recent years, the demand of people on skin care is increasing, the research on oil control and anti-inflammatory is gradually in progress, and various raw materials with oil control and anti-inflammatory activities are continuously developed aiming at various mechanisms of skin oil control and anti-inflammatory. Along with the continuous development of science, active ingredients in natural plants and the efficacy thereof are continuously excavated. For example, biota orientalis, sabina chinensis, juniper and selaginella tamariscina, which are collected in the "catalog of used cosmetic raw materials" 2021. According to reports, all four plants have various flavonoid components, and particularly all contain quercetin and quercitrin. Pharmacological studies have shown that their extracts have anti-inflammatory, antioxidant and bacteriostatic effects. At present, the active ingredients in the natural plants are obtained by a solvent extraction method, and the active ingredients exist in the traditional extraction methodThe fraction yield is low, the energy consumption is high, and the effect of the obtained extract is not obvious; some manufacturers use microwave extraction and supercritical CO ₂ Extraction is carried out, but equipment requirements are high, and the cost is high.

Therefore, how to increase the active ingredient in plant extracts is a technical problem to be solved in the art.

Disclosure of Invention

The present invention aims to solve at least one of the technical problems in the prior art described above. Therefore, the invention provides the plant ferment capable of controlling and relieving the oil, and the preparation method and the application thereof, and the plant ferment has high content of active ingredients of total flavone and quercetin, thereby having remarkable effect of controlling and relieving the oil.

In a first aspect, the present invention provides a plant ferment, which is prepared from the following raw materials: plant material, cow milk, glycerol and fermentation strains; the plant material comprises at least one of biota orientalis, juniper leaf and selaginella tamariscina leaf; the fermentation strain comprises at least one of lactobacillus plantarum, lactobacillus lactis and bifidobacterium longum.

According to some embodiments of the invention, the plant material, the cow's milk and the glycerol are used in a ratio of (0.01-50 g): (0.01-50 g): 1g. Further, the plant material, the cow milk and the glycerin are used in an amount ratio of (0.01-10 g): (0.01-10 g): 1g. Still further, the plant material, the cow's milk and the glycerin are used in an amount ratio of (0.1-1 g): (0.1-1 g): 1g.

According to some embodiments of the invention, the plant ferment preparation feedstock further comprises water.

According to some embodiments of the invention, the ratio of the glycerol to the water is 1g: (1-1000 g). Further, the ratio of the glycerin to the water was 1g: (1-100 g). Still further, the ratio of the amount of the glycerin to the amount of the water was 1g: (10-50 g).

According to some embodiments of the invention, the plant ferment contains an active ingredient comprising > 1mg/mL total flavonoids, and > 80 μg/mL quercetin. Further, the active ingredient comprises 1-3mg/mL of total flavonoids and 80-150 mug/mL of quercetin.

According to some embodiments of the invention, the concentration of the fermentation strain is 10 ⁵ -10 ¹⁰ CFU/mL. Further, the concentration of the fermentation strain is 10 ⁸ -10 ¹⁰ CFU/mL。

A second aspect of the present invention provides a method for producing the above plant fermented product, comprising the steps of:

mixing the plant raw material, the cow milk and the glycerol, and homogenizing under high pressure to obtain a plant composite liquid;

mixing the plant composite liquid with the fermentation strain, and fermenting to obtain the plant fermentation product;

the pressure of the high-pressure homogenization is 20-100MPa.

According to some embodiments of the invention, the high pressure homogenization is at a pressure of 50-100MPa.

According to some embodiments of the invention, the temperature of the high pressure homogenization is 25-50 ℃ and the time of the high pressure homogenization is 5-40min. Further, the temperature of the high-pressure homogenization is 30-50 ℃, and the time of the high-pressure homogenization is 15-20min.

According to some embodiments of the invention, a culture medium is also added before the fermentation is performed; the dosage ratio of the plant composite liquid, the fermentation strain and the culture medium is (1-500 g): (0.01-10 mL): 1g. Further, the plant complex, the fermentation strain and the culture medium are used in an amount ratio of (1-100 g): (0.1-5 mL): 1g. Further, the plant composite liquid, the fermentation strain and the culture medium are used in an amount ratio of (20-80 g): (0.1-1 mL): 1g.

According to some embodiments of the invention, the medium is MRS medium.

According to some embodiments of the invention, the fermentation is performed at a temperature of 20-45 ℃ for a time of 12-120 hours. Further, the temperature of the fermentation is 30-40 ℃, and the time of the fermentation is 72-120h.

According to some embodiments of the invention, after the fermentation, the fermentation broth is sterilized.

According to some embodiments of the invention, the temperature of the sterilization is 100-150 ℃ and the time of the sterilization is 10-30min.

According to some embodiments of the invention, prior to the high pressure homogenizing, further comprising mixing water with the plant material, the cow milk, the glycerin.

A third aspect of the present invention provides the use of the above plant ferment for the preparation of a cosmetic or external skin preparation.

A fourth aspect of the present invention provides a cosmetic comprising the plant ferment described above.

According to some embodiments of the invention, the cosmetic further comprises a cosmetically acceptable adjuvant.

According to some embodiments of the invention, the cosmetically acceptable adjuvant includes at least one of an emulsifier, a humectant, a thickener, a preservative.

According to some embodiments of the invention, the plant ferment is added to the cosmetic in an amount of 0.01-80wt%. Further, the plant ferment is added into the cosmetic in an amount of 0.05-20wt%. Further, the plant ferment is added into the cosmetic in an amount of 0.1-10wt%.

According to some embodiments of the invention, the cosmetic comprises a shampoo, a body wash, or a facial cleanser.

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

the invention adopts specific plant raw materials, fermentation strain, cow milk and glycerin as preparation raw materials, and the plant fermentation product prepared by fermentation has high content of active ingredients of total flavone and quercetin, wherein the content of total flavone reaches 1-3mg/mL, the content of quercetin reaches 80-150 mug/mL, and the invention has remarkable oil control and relief effects.

According to the invention, cow milk and glycerol are added in the high-pressure homogenization process, so that plant tissue cells are fully crushed, the dissolution rate of active ingredients is improved, the method is organically combined with the directional microorganism fermentation technology, a specific strain is adopted for fermentation, the plant composite stock solution obtained by high-pressure homogenization is used as a subsequent fermentation matrix, and the contents of total flavonoids and quercetin in the plant fermentation product are finally improved through the bioconversion capability of microorganisms, so that the plant fermentation product has a remarkable oil control and relief effect.

Detailed Description

In order to make the technical solutions of the present invention more apparent to those skilled in the art, the following examples will be presented. It should be noted that the following examples do not limit the scope of the invention.

The starting materials, reagents or apparatus used in the following examples are all available from conventional commercial sources or may be obtained by methods known in the art unless otherwise specified.

Example 1

The preparation raw materials of the plant ferment comprise: biota orientalis, cow milk, glycerol and lactobacillus plantarum bacterial liquid.

The preparation method of the plant fermentation product comprises the following steps:

1) Mixing 10g of cacumen biotae, 20g of cow milk, 20g of glycerol and 360g of deionized water, adding into a high-pressure homogenizer for high-pressure homogenization, wherein the homogenization temperature is 40 ℃, the homogenization pressure is 80MPa, and the homogenization time is 15min to obtain a plant composite liquid;

2) 200g of plant complex solution and 2mL of lactobacillus plantarum bacterial solution (concentration is 10) ⁸ CFU/mL) and 4g of MRS medium, fermenting at 30 ℃ for 72 hours, sterilizing the fermentation broth at 121 ℃ for 20 minutes after the fermentation is finished, centrifuging the sterilized fermentation broth at 6000rpm/min for 20 minutes, and taking the supernatant after centrifugation to obtain a plant fermentation product.

Example 2

The preparation raw materials of the plant ferment comprise: the preparation method comprises the steps of cacumen biotae, cow milk, glycerol and lactobacillus plantarum bacterial liquid.

Example 3

The preparation raw materials of the plant ferment comprise: juniper leaf, cow milk, glycerol and lactobacillus plantarum bacterial liquid.

1) Mixing 10g of juniper leaves, 20g of cow milk, 20g of glycerol and 360g of deionized water, and then adding into a high-pressure homogenizer for high-pressure homogenization, wherein the homogenization temperature is 40 ℃, the homogenization pressure is 80MPa, and the homogenization time is 15min to obtain a plant composite liquid;

Example 4

The preparation raw materials of the plant ferment comprise: herba Selaginellae, milk, glycerol and lactobacillus plantarum bacterial solution.

1) Mixing herba Selaginellae 10g, cow milk 20g, glycerol 20g, and deionized water 360g, and homogenizing under high pressure at 40deg.C under 80MPa for 15min to obtain plant composite solution;

2) 200g of plant complex solution and 2mL of lactobacillus plantarum bacterial solution (concentration is 10) ⁸ CFU/mL) and 4g MRS culture medium, fermenting at 30deg.C for 72 hr, and fermenting at 121Sterilizing at 6000rpm/min for 20min, centrifuging for 20min, and collecting supernatant to obtain plant fermented product.

Example 5

1) Mixing 5g of cacumen biotae, 20g of cow milk, 20g of glycerol and 360g of deionized water, and adding into a high-pressure homogenizer for high-pressure homogenization at 40 ℃ under 80MPa for 20min to obtain plant composite liquid;

Example 6

The preparation raw materials of the plant ferment comprise: juniper leaf, selaginella leaf, cow milk, glycerol and lactobacillus plantarum bacterial liquid.

1) Mixing 5g of juniper leaves, 5g of selaginella, 20g of cow milk, 20g of glycerol and 360g of deionized water, and then adding into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40 ℃ under a homogenization pressure of 80MPa for 20min to obtain a plant composite liquid;

Example 7

The preparation raw materials of the plant ferment comprise: biota orientalis, juniper leaf, selaginella tamariscina leaf, cow milk, glycerol and lactobacillus plantarum bacterial liquid.

1) Mixing 2.5g of cacumen Platycladi, 2.5g of folium Juniperi, 2.5g of herba Selaginellae, 20g of cow milk, 20g of glycerol and 360g of deionized water, and adding into a high-pressure homogenizer for high-pressure homogenization at 40deg.C under 80MPa for 20min to obtain plant composite liquid;

Example 8

The preparation raw materials of the plant ferment comprise: biota orientalis, cow milk, glycerol and lactobacillus bacteria liquid.

2) 200g of the plant complex solution and 2mL of lactobacillus strain solution (concentration 10) ⁸ CFU/mL) and 4g of MRS medium, fermenting at 33 ℃ for 72 hours, sterilizing the fermentation broth at 121 ℃ for 20 minutes after the fermentation is finished, centrifuging the sterilized fermentation broth at 6000rpm/min for 20 minutes, and taking the supernatant after centrifugation to obtain a plant fermentation product.

Example 9

The preparation raw materials of the plant ferment comprise: the preparation method comprises the steps of cacumen biotae, cow milk, glycerol and lactobacillus bacteria liquid.

Example 10

The preparation raw materials of the plant ferment comprise: juniper leaf, cow milk, glycerol and lactobacillus bacteria liquid.

Example 11

The preparation raw materials of the plant ferment comprise: herba Selaginellae, milk, glycerol and lactobacillus bacteria solution.

Example 12

Example 13

The preparation raw materials of the plant ferment comprise: juniper leaf, selaginella leaf, cow milk, glycerol and lactobacillus bacteria liquid.

Example 14

The preparation raw materials of the plant ferment comprise: biota orientalis, juniper leaf, selaginella tamariscina leaf, cow milk, glycerol and lactobacillus bacteria liquid.

Example 15

The preparation raw materials of the plant ferment comprise: biota orientalis, cow milk, glycerol and bifidobacterium longum bacterial liquid.

2) 200g of the plant complex solution and 2mL of bifidobacterium longum bacterial solution (concentration is 10) ⁸ CFU/mL) and 4g of MRS medium, fermenting at 37 ℃ for 72 hours, sterilizing the fermentation broth at 121 ℃ for 20 minutes after the fermentation is finished, centrifuging the sterilized fermentation broth at 6000rpm/min for 20 minutes, and taking the supernatant after centrifugation to obtain a plant fermentation product.

Example 16

The preparation raw materials of the plant ferment comprise: the preparation method comprises the steps of cacumen biotae, cow milk, glycerol and bifidobacterium longum bacterial liquid.

Example 17

The preparation raw materials of the plant ferment comprise: juniper leaf, cow milk, glycerin and bifidobacterium longum bacterial liquid.

Example 18

The preparation raw materials of the plant ferment comprise: herba Selaginellae, milk, glycerol and Bifidobacterium longum bacterial liquid.

Example 19

Example 20

The preparation raw materials of the plant ferment comprise: juniper leaf, selaginella leaf, cow milk, glycerol and bifidobacterium longum bacterial liquid.

2) 200g of the plant complex solution and 2mL of bifidobacterium longum bacterial solution (concentration is 10) ⁸ CFU/mL) and 4g MRS culture medium, fermenting at 37deg.C for 72 hr, sterilizing the fermentation broth at 121deg.C for 20min after fermentation, and sterilizing the fermentation broth at 6000rpm/mCentrifuging for 20min, and collecting supernatant to obtain plant fermentation product.

Example 21

The preparation raw materials of the plant ferment comprise: biota orientalis, juniper leaf, selaginella tamariscina leaf, cow milk, glycerol and bifidobacterium longum bacterial liquid.

Comparative example 1 (differing from example 7 only in the conventional water decoction)

A method for preparing a compound extract, comprising the steps of:

mixing 2.5g of biota orientalis, 2.5g of hinoki leaf, 2.5g of juniper leaf, 2.5g of selaginella tamariscina leaf, 20g of cow milk, 20g of glycerol and 360g of deionized water, heating and extracting for 1h at 100 ℃, filtering, and collecting filtrate to obtain a compound extract.

Comparative example 2

This comparative example is different from example 7 only in that the cow's milk and glycerin in step 1) were replaced with deionized water, and the amount of addition was the same, and other raw materials, amounts of addition, and preparation methods were the same as example 7.

Comparative example 3

This comparative example is different from example 7 only in that the cow's milk in step 1) was replaced with honey and the amount added was the same, and other raw materials, amounts added and preparation methods were the same as example 7.

Comparative example 4

This comparative example is different from example 7 only in that the glycerin in step 1) was replaced with deionized water and the amount added was the same, and other raw materials, amounts added, and preparation methods were the same as in example 7.

Comparative example 5

This comparative example is different from example 7 only in that the cow's milk in step 1) was replaced with lecithin and the amount added was the same, and other raw materials, amounts added and preparation methods were the same as in example 7.

Comparative example 6

This comparative example is different from example 7 only in that glycerin is replaced with butanediol in step 1), and the amount of addition is the same, and other raw materials, the amount of addition, and the preparation method are the same as example 7.

Comparative example 7

This comparative example is different from example 7 only in that the lactobacillus plantarum bacterial solution in step 2) is replaced with deionized water, and the addition amount is the same, and other raw materials, addition amounts and preparation methods are the same as those of example 7.

Comparative example 8

This comparative example is different from example 7 only in that the lactobacillus plantarum bacterial liquid in step 2) is replaced with the monascus bacterial liquid, and the addition amount is the same, and other raw materials, addition amounts and preparation methods are the same as those of example 7.

Test example 1: determination of total flavone content

This test example tests the total flavone content of the plant ferments of examples 1 to 21 described above, and the plant ferments or complex extracts (hereinafter referred to as "samples") of comparative examples 1 to 8. The specific test method is as follows:

and (3) adjusting the total flavone concentration of the sample solution to a measurement range, accurately sucking 2.0mL, placing in a dry clean test tube, adding 0.2mL of 5% sodium nitrite solution, shaking uniformly, placing for 5min, adding 0.2mL of 10% aluminum nitrate solution, shaking uniformly, placing for 6min, adding 1.0mL of 5% sodium hydroxide solution, adding a solvent to a scale, and shaking uniformly to obtain a sample test solution. 3 replicates were made for each concentration. The sample solution was rapidly assayed for absorbance at 510 nm. And calculating the concentration of total flavonoids according to the standard curve of the rutin solution and the absorbance value of the sample solution, wherein the total flavonoids content in the sample is represented by rutin.

Test example 2: content determination of Quercetin

This test example tests the content of quercetin in the plant fermented products of the above examples 1 to 21, and the plant fermented products or composite extracts (hereinafter referred to as "samples") of comparative examples 1 to 8. The specific test method is as follows:

test sample preparation: taking 5mL of a sample, adding 5mL of chromatographic grade acetonitrile, mixing uniformly, centrifuging at 6000rpm/min for 20min, taking supernatant, and filtering with a 0.45 mu m filter membrane to obtain a test sample.

Chromatographic conditions: using a Agilent ZORBAX SB-C18 column (4.6mm.times.250 mm,5 μm); gradient elution was performed with 0.1% phosphoric acid (A) -acetonitrile (B) as mobile phase (0 min,15% B;10min,30% B;19min,30% B), flow rate 1.0mL/min; a detection wavelength of 254nm; column temperature 25 ℃.

The test results of test examples 1 and 2 are shown in Table 1.

TABLE 1

As can be seen from Table 1, the total flavone content in the samples of examples 1 to 21 was greatly improved as compared with comparative examples 1 to 8, probably because microorganisms were able to efficiently separate plant tissues, making the active ingredient more easily eluted; meanwhile, cow milk and glycerol are added in pretreatment, which is favorable for dissolving low-polarity flavonoid components, so that the content of total flavonoids in a sample is improved. In addition, the content of quercetin is also significantly increased, probably due to the conversion of other components by active enzymes present in the microorganism.

Test example 3: cell scratch assay

This test example was conducted on the plant fermented products of the above examples 1 to 21, and the plant fermented products or the composite extracts (hereinafter referred to as "samples") of comparative examples 1 to 8, for evaluation of cell repair ability and for test of influence on cell migration movement. The specific test method is as follows:

the pen was used to draw lines evenly across the back of the 6-well plate with about 0.5-1cm spacing between lines. Spreading human immortalized epidermal cells (HaCaT) in the hole, spreading the next day, spreading the cells in the hole, drawing the cells by using a gun head according to the transverse line drawn in advance by a 6-hole plate, washing the cells with PBS for 3 times, removing the drawn cells, adding a cell culture medium containing 1% (w/w) sample, and adding 5% CO at 37deg.C ₂ Culturing for 24h in an incubator, photographing, and counting the scratch healing proportion of 24h. Deionized water was used instead of the sample as a blank. The evaluation results are shown in Table 2.

TABLE 2

As can be seen from Table 2, the samples of examples 1 to 21 are more effective in improving the migration and repair ability of cells than those of comparative examples 1 to 8, which indicates that the plant ferment obtained by the preparation method of the present invention has excellent repair effect on epidermal cells. The components affecting the cell viability may exist in the obtained product without cow milk, glycerol treatment and microbial fermentation, resulting in a significant reduction in the repair efficacy of the product on epidermal cells.

Test example 4: effect on IL-6 and TNF- α content in HaCaT cells after UV irradiation

Human epidermal keratinocytes were seeded into 6-well plates at 37℃and 5% CO ₂ Is incubated overnight. When the cell plating rate in the 6-hole plate reaches 50% -60%, adding a cell culture medium into the blank group, and performing no other treatment; adding cell culture medium into the model group, adding cell culture medium containing 1% (w/w) sample into the sample group, pretreating for 1 hr,respectively performing UVB (medium wave ultraviolet) irradiation for 40min with radiation intensity of 30mJ/cm ² . After treatment at 37℃and 5% CO ₂ The culture was continued for 24 hours. After the cultivation, the IL-6 and TNF-alpha contents were determined according to the methods of operation of the Human IL-6ELISA Kit (Biyun day) and the Human TNF-alpha ELISA Kit (Biyun day), respectively. The test results are shown in Table 3.

TABLE 3 Table 3

The results in Table 3 show that the plant ferments prepared in examples 1-21 are more effective in reducing the levels of inflammatory factors IL-6 and TNF- α produced by HaCaT cells after UV irradiation, and exhibit anti-inflammatory and soothing activities, as compared to comparative examples 1-8.

Test example 5: action against Staphylococcus aureus and Malachite's bacteria

Preparing a bacterial suspension: staphylococcus aureus (ATCC 6538) and malassezia furfur (ATCC 14521) were selected as test bacteria. Among them, malassezia furfur is a conditional pathogenic fungus, and is propagated in large quantity under the condition of vigorous secretion of sebaceous glands, and can cause folliculitis, tinea versicolor and the like, and is known to be closely related to dandruff production and scalp health; staphylococcus aureus is also a common pathogenic bacterium of the skin and can often cause papules or pustules in the skin. Preparing test bacteria into bacterial suspension with PBS with concentration of 5×10 ⁶ CFU/mL。

And (3) bacteriostasis circle test: the oxford cup method is adopted for testing the inhibition zone. Sucking 100 mu L of bacterial suspension on the surface of the solidified culture medium, uniformly smearing the bacterial suspension on the surface of the solidified culture medium by using a sterile coating rod, covering a plate, and standing at 20 ℃ for 10min; and finally, lightly inverting the sterile oxford cup on the surface of the culture medium, dripping sample solutions with different mass fractions until the liquid level is parallel to the surface of the oxford cup, and placing the mixture into an incubator for culturing for 18 hours at 37 ℃. Reference to the determination standard of the result of the inhibition zone in the enterprise standard "in vitro inhibition of bacterial and fungal test operation Instructions (drug sensitivity test) of the microbial antimicrobial evaluation value": the zone of inhibition is extremely sensitive with a diameter >20mm, expressed as++ + +;15-20mm is highly sensitive, expressed as++; 10-15mm is moderately sensitive, expressed as++; <10mm is low sensitivity, expressed as +;0mm is no antibacterial effect, expressed as-. The test results are shown in Table 4.

TABLE 4 Table 4

From the test results shown in Table 4, it is understood that the samples of examples 1 to 21 have inhibitory effects on Staphylococcus aureus and Malassezia furfur at a mass fraction of 5% or 1%. The inhibition of Staphylococcus aureus and Malassezia furfur was stronger at the same concentration than the samples of comparative examples 1 to 8. The samples of examples 1-21 have good inhibitory effects on pathogenic bacteria, demonstrating that they can be used to maintain normal healthy skin micro-ecology.

Test example 6: erythrocyte hemolysis evaluation

This test example was conducted to evaluate erythrocyte hemolysis of the plant fermented products of examples 1 to 21 described above, and the plant fermented products or the composite extracts (hereinafter referred to as "samples") of comparative examples 1 to 8. The basic principle of the erythrocyte hemolysis experiment is to evaluate the damage of chemicals to ocular tissue cells by measuring the leaching amount and the denaturation degree of hemoglobin. The experiment is based on the test method and grading standard of European alternative method verification center. Where HD50 is the concentration of the sample at which 50% of the erythrocytes are hemolyzed, DI is the protein denaturation index, and L/D is the ratio of HD50 to DI. The evaluation criteria are shown in Table 5, and the evaluation results are shown in Table 6.

TABLE 5

L/D	Grading
		L/D>100	No irritation
10<L/D≤100	Microstimulation
		1<L/D≤10	Mild irritation
0.1<L/D≤1	Toxicity irritation
		L/D≤0.1	Severe irritation

TABLE 6

As can be seen from Table 6, the plant ferments or complex extracts of examples 1-21 and comparative examples 1-7 were relatively mild starting materials, except that comparative example 8 was slightly irritating, and the materials used before and after fermentation were non-irritating.

Application example 1

The present application example provides a shampoo comprising the plant ferment of example 7. The formulation of the composition is shown in Table 7 in percentage by mass. The shampoo of application example 1 was prepared according to the conventional preparation method in the art.

TABLE 7

Comparative example 1 was used

Application comparative example 1 differs from application example 1 in that the plant fermented product of example 7 was replaced with the composite extract of comparative example 1 by application comparative example 1, and other ingredients, amounts and preparation methods were the same as those of application example 1.

Test example 7: evaluation of anti-dandruff and soothing efficacy of shampoo

60 volunteers were selected and randomly divided into 2 groups, aged between 20-50 years, and male and female halves. Group 1 used 5g of shampoo of application example 1 on day 1, day 3, day 5, and day 7, respectively; group 2 used 5g of shampoo of application comparative example 1 on day 1, day 3, day 5, and day 7, respectively. On day 8, subjects were given a return visit for effect, and subjects scored for use sensation. The scoring standard adopts 10 scores, and the higher the score, the better the use feeling. Improvement of scales (0 is no improvement, 10 is obvious improvement); improvement of itching (no improvement in 0 and significant improvement in 10); the overall feeling of use (0 is very poor and 10 is very good). The test results are shown in Table 8.

Test example 8: evaluation of shampoo oil control efficacy

20 volunteers with vigorous scalp grease secretion were recruited, aged between 20-30 years. If the volunteer avoids the physiological cycle and life work and rest law for the female, the volunteer avoids intense exercise, insolation, staying up at night and the like as much as possible. The hair was not rinsed the evening the day before the test. The volunteers were sitting still for 30min in a room at a temperature of 20-22 ℃ and a humidity of 40% -65% before testing, during which they were unable to drink water and beverages. The test part is exposed, placed in a test state and kept relaxed. The test time points are before, immediately after, 6 hours and 8 hours. The test site was taken as a forehead-top 5cm area and a 1cm x 1cm square was drawn on the test area with a skin pen. When the sample is used, half-head hair washing is adopted. The hair line is divided into middle parts, half parts are blank control, and the hair is washed by clean water. The other half is the sample area, and the test sample is used for washing hair. The tape strip is bent into a circular ring to be contacted with the scalp, the lipoid compound (grease) in the scalp changes the transparency of the test tape, and then the test tape strip is flattened and then put into a slide block of a scalp grease tester, and the slide block is pushed into the instrument. The light transmittance distribution curve on the adhesive tape can be obtained through testing by a photoelectric tube in the instrument, the peak value of the light transmittance distribution curve represents the content of lipid compounds, namely grease, in the scalp, and the annular top end of the adhesive tape is just the part which is in full contact with the scalp during testing. Calculation of scalp fat mean difference, scalp fat mean difference of blank and sample groups at different time periods = scalp fat content of blank-scalp fat content of sample group. The test results are shown in Table 9.

TABLE 8

Sample of	Anti-dandruff effect	Anti-itching effect	Feel in use as a whole
				Example 7	8.6	8.0	8.5
Comparative example 1	6.2	6.4	7.3

TABLE 9

As is clear from Table 8, the shampoo of application example 1, to which the plant fermentation product of example 7 was added, had remarkable anti-dandruff and anti-itching effects after use, and the overall feel was also greatly improved, as compared with the shampoo of application comparative example 1. The plant fermentate of example 7 was shown to inhibit not only IL-6 and TNF- α production, but also Staphylococcus aureus and Malassezia furfur, and the anti-dandruff and anti-itching effects of the shampoo correlated with the anti-inflammatory and antibacterial efficacy of the raw materials.

In addition, as is clear from table 9, shampoo added with the plant ferment of example 7 slows down the secretion of scalp grease, and achieves the oil control effect.

While the preferred embodiments of the present invention have been illustrated and described, the present invention is not limited to the embodiments, and various equivalent modifications and substitutions can be made by one skilled in the art without departing from the spirit of the present invention, and these are intended to be included in the scope of the present invention as defined in the appended claims.

Claims

1. A plant ferment, characterized in that the preparation raw materials of the plant ferment comprise: plant material, cow milk, glycerol and fermentation strains; the plant material comprises at least one of biota orientalis, juniper leaf and selaginella tamariscina leaf; the fermentation strain comprises at least one of lactobacillus plantarum, lactobacillus lactis and bifidobacterium longum.

2. The plant ferment according to claim 1, characterized in that the ratio of the amounts of the plant raw material, the cow's milk and the glycerol is (0.01-50 g): (0.01-50 g): 1g.

3. The plant ferment of claim 1, wherein the plant ferment contains an active ingredient comprising > 1mg/mL total flavonoids, and > 80 μg/mL quercetin.

4. The plant ferment of claim 1, wherein the concentration of the ferment strain is 10 ⁵ -10 ¹⁰ CFU/mL。

5. A method for producing a plant ferment according to any one of claims 1 to 4, characterized by comprising the steps of:

the pressure of the high-pressure homogenization is 20-100MPa.

6. The method according to claim 5, wherein a medium is further added before the fermentation is performed; the dosage ratio of the plant composite liquid, the fermentation strain and the culture medium is (1-500 g): (0.01-10 mL): 1g.

7. The method according to claim 5, wherein the fermentation is carried out at a temperature of 20 to 45 ℃ for a period of 12 to 120 hours.

8. Use of the plant ferment according to any one of claims 1 to 4 for the preparation of a cosmetic or external skin preparation.

9. A cosmetic comprising the plant ferment of any one of claims 1 to 4.

10. The cosmetic product according to claim 9, wherein the cosmetic product comprises a shampoo, a body wash or a facial cleanser.