Traditional Chinese medicine fermentation extracting solution as well as preparation method and application thereof

Abstract

The invention provides a traditional Chinese medicine fermentation extracting solution and a preparation method and application thereof, and relates to the technical field of traditional Chinese medicine cosmetics. A traditional Chinese medicine composition which is composed of sophora flower bud, clove, honeysuckle, citron, astragalus membranaceus, houttuynia cordata, turmeric, emblic leafflower fruit, rehmannia and roselle and has an anti-aging effect is reasonably combined, and the traditional Chinese medicine fermented extract is obtained after the traditional Chinese medicine composition is fermented for multiple times by utilizing compound probiotics. The traditional Chinese medicine fermentation extract has a strong free radical scavenging effect, has an obvious improvement effect on cell activity, can obviously inhibit matrix protease activity, can obviously improve MDA content in serum of an aging mouse model and indexes of SOD, CAT, GSH-Px activity and the like, and has obvious anti-oxidation and anti-aging effects. The traditional Chinese medicine fermentation extracting solution provided by the invention can be used for preparing anti-aging cosmetics or functional foods, is safe and non-irritant when being used for human skin, and has an obvious improvement effect on the elasticity, wrinkles and smoothness of the human skin.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine cosmetics, and specifically relates to a traditional Chinese medicine fermentation extract and a preparation method thereof, and also relates to the application of the traditional Chinese medicine fermentation extract in the field of cosmetics. Background Art

[0002] Traditional Chinese medicine cosmetics are favored by researchers and consumers because their active ingredients come from the natural essence of plants themselves. Different from other cosmetics, traditional Chinese medicine cosmetics are guided by traditional Chinese medicine theory and use natural plants as raw materials, with obvious functionality, safety and reliability; and traditional Chinese medicine has always been concerned for its advantages such as purity, mildness, and low toxicity and side effects. The multiple active ingredients in its medicinal materials have multiple effects, can be absorbed by the human body, and can effectively address various skin problems.

[0003] Although there are currently many traditional Chinese medicines with low toxicity that also have good antioxidant, anti-aging, anti-inflammatory, acne-removing and other effects, the traditional processing procedures are complex and strict, the utilization rate of medicinal effects is low, the medicinal effects largely depend on the stability of the processing techniques, and there is a lack of a large-scale and stable industrial production process, making it difficult to expand the market supply. And as the ancients said: "Every medicine has some toxicity." The earliest medical monograph in China, "Huangdi Neijing", is very particular about how to use medicine, and classifies medicine into highly toxic, moderately toxic, slightly toxic, and non-toxic. For treating diseases, it is required that for highly toxic medicine, six-tenths of its toxicity be removed; for moderately toxic medicine, seven-tenths of its toxicity be removed; for slightly toxic medicine, eight-tenths of its toxicity be removed; for non-toxic medicine, nine-tenths of its toxicity be removed. Nowadays, many people think that most traditional Chinese medicines are derived from natural animals, plants and pure traditional Chinese medicine preparations, and are milder and safer in medicinal properties than chemical drugs, and always think that there will be no drug toxicity and side effects. In fact, this is not the case. If used arbitrarily and misused, drug toxicity and side effects will also occur.

[0004] At present, many functional cosmetics are prepared from traditional Chinese medicine extracts. However, there are problems such as low content of effective active ingredients in traditional Chinese medicine, low bioavailability of active ingredients, small absorption degree of human skin, and poor improvement of skin condition. In order to improve the effect of traditional Chinese medicine cosmetics, existing research has shown that fermenting traditional Chinese medicine extracts by microbial fermentation, using microorganisms to convert raw materials into required products through specific metabolic pathways under suitable conditions, this process can improve the activity of each component, thereby improving the effect. During the treatment of traditional Chinese medicine raw materials with probiotics, many enzyme substances will be produced, such as cellulase, hemicellulase, pectinase, etc. These enzymes can accelerate the enzymatic hydrolysis of plant cell walls, promote the release of active ingredients, and further decompose and convert the beneficial ingredients that are difficult to absorb in traditional Chinese medicine into small molecules that are easy to be absorbed by the human body, so as to make the effective components of traditional Chinese medicine be utilized to a greater extent. At the same time, these enzyme substances can interact with certain special components in traditional Chinese medicine and modify or transform them to promote the formation of new effective substances and reduce the content of toxic components in traditional Chinese medicine. Moreover, traditional Chinese medicine materials and probiotics from natural sources are safer and more acceptable to the public. Therefore, applying fermented traditional Chinese medicine to cosmetics and functional foods has good prospects. At present, the traditional Chinese medicine cosmetics on the market have problems such as overly extensive functions, unclear effects, and no targeted solutions to skin problems. Based on this, the present invention aims to provide a traditional Chinese medicine fermented extract with obvious anti-aging effects and its application in the field of cosmetics technology. Summary of the Invention

[0005] (1) Technical problems to be solved

[0006] In view of the deficiencies of the prior art, the present invention provides a traditional Chinese medicine fermented extract with anti-aging effects, its preparation method, and the application of the fermented extract in the field of cosmetics technology.

[0007] (2) Technical solutions

[0008] To achieve the above objectives, the present invention is realized through the following technical solutions:

[0009] In the first aspect, the present invention provides a traditional Chinese medicine fermented extract, which is prepared by co-fermenting a traditional Chinese medicine composition and compound probiotics.

[0010] Specifically, the traditional Chinese medicine composition is composed of the following raw materials in parts by weight: 0-32 parts of sophora flower buds, 0-32 parts of cloves, 0-32 parts of honeysuckle flowers, 0-32 parts of citrons, 0-32 parts of astragalus membranaceus, 0-32 parts of houttuynia cordata, 0-32 parts of turmeric, 0-32 parts of phyllanthus emblica, 0-32 parts of rehmannia glutinosa, 0-32 parts of roselle.

[0011] Specifically, the traditional Chinese medicine composition is composed of the following raw materials in parts by weight: 32 parts of sophora flower buds, 32 parts of cloves, 32 parts of honeysuckle flowers, 8 parts of citrons, 16 parts of astragalus membranaceus, 8 parts of houttuynia cordata, 2 parts of turmeric, 8 parts of phyllanthus emblica, 4 parts of rehmannia glutinosa, and 4 parts of roselle.

[0012] Specifically, the traditional Chinese medicine composition is composed of the following raw materials in parts by weight: 32 parts of sophora flower buds, 16 parts of cloves, 32 parts of honeysuckle flowers, 8 parts of citrons, 2 parts of astragalus membranaceus, 2 parts of houttuynia cordata, 2 parts of turmeric, 8 parts of phyllanthus emblica, 8 parts of rehmannia glutinosa, and 8 parts of roselle.

[0013] Specifically, the traditional Chinese medicine composition is composed of the following raw materials in parts by weight: 32 parts of sophora flower buds, 32 parts of cloves, 32 parts of honeysuckle flowers, 8 parts of citrons, 2 parts of astragalus membranaceus, 2 parts of houttuynia cordata, 2 parts of turmeric, 32 parts of phyllanthus emblica, 32 parts of rehmannia glutinosa, and 16 parts of roselle.

[0014] Specifically, the compound probiotics are one or more of yeast, bacillus coagulans, fungi, and lactobacillus.

[0015] Specifically, the compound probiotics are composed of eurotium cristatum: bacillus coagulans: yeast with a mass ratio of 4 - 2:2 - 0.5:1 - 0.1.

[0016] Specifically, the compound probiotics are composed of eurotium cristatum: bacillus coagulans: yeast with a mass ratio of 4:2:1.

[0017] In a second aspect, the present invention also provides a preparation method of a traditional Chinese medicine fermentation extract. The traditional Chinese medicine fermentation extract is prepared according to the following steps:

[0018] (1) Weigh each traditional Chinese medicine raw material according to the above parts by weight, wash, dry and crush it, then transfer it to a flask, add 5 - 20 times the weight of pure water, add the compound probiotics with a volume fraction of 5 - 35% of the pure water volume in a laminar flow hood, and carry out a fermentation reaction at 28 - 42 °C for 2 - 10 days to obtain a traditional Chinese medicine fermentation product; this step is repeated 1 - 3 times;

[0019] (2) Place the traditional Chinese medicine fermentation product in a decocting machine, add 10 - 20 times the pure water according to the weight of the traditional Chinese medicine composition, decoct at a gentle boil for 1.2 - 2 h, then let it stand for 2 - 4 h, take the supernatant and filter it, and concentrate it to a mass fraction of about 4 - 6 times the amount of traditional Chinese medicine input under the conditions of a vacuum negative pressure of -0.07 to -0.1 MPa and a temperature of 50 - 60 °C to obtain a crude traditional Chinese medicine fermentation extract;

[0020] (3) The crude traditional Chinese medicine fermentation extract is subjected to precipitation, centrifugation, decolorization, and desalting to obtain the final traditional Chinese medicine fermentation extract.

[0021] Thirdly, the present invention also provides an application of a traditional Chinese medicine fermentation extract, and the traditional Chinese medicine fermentation extract is used for preparing anti-aging cosmetics or functional foods; the traditional Chinese medicine combination is incorporated into the anti-aging cosmetics or functional foods at a ratio of 0.1% to 50% by weight.

[0022] (III) Beneficial effects

[0023] The present invention rationally formulates a traditional Chinese medicine composition with anti-aging effects composed of sophora japonica, cloves, honeysuckle, citron, astragalus membranaceus, houttuynia cordata, turmeric, phyllanthus emblica, rehmannia glutinosa, and roselle, and uses compound probiotics to ferment the above traditional Chinese medicine composition multiple times to obtain a traditional Chinese medicine fermentation extract. The traditional Chinese medicine fermentation extract has a strong free radical scavenging effect, significantly improves cell viability, can significantly inhibit the activity of matrix protease, can significantly improve the content of MDA and the activities of SOD, CAT, and GSH-Px in the serum of the aging mouse model, and has significant antioxidant and anti-aging effects. The traditional Chinese medicine fermentation extract provided by the present invention can be used for the preparation of anti-aging cosmetics or functional foods, and the traditional Chinese medicine fermentation extract is safe and non-irritating to human skin and has an obvious improvement effect on the elasticity, wrinkles, and smoothness of human skin. Description of the drawings

[0024] Figure 1 Cell viability of the samples of Examples 1-4 and Comparative Examples 1-12, *P<0.05, **P<0.01.

[0025] Figure 2 MMP-1 inhibition rate of the samples of Examples 1-4 and Comparative Examples 1-12, *P<0.05, **P<0.01.

[0026] Figure 3 Content of MDA (A) and relative activities of SOD (B), CAT (C), and GSH-Px (D) in mouse serum; *P<0.05, **P<0.01.

[0027] Figure 4 Change rates of skin elasticity (A), wrinkles (B), and smoothness (C) after using the samples of Examples 1-4; *P<0.05, **P<0.01. Detailed implementation manners

[0028] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are some, but not all, of the embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0029] Example 1

[0030] A traditional Chinese medicine fermentation extract is prepared according to the following steps:

[0031] Weigh 32 g of sophora japonica flower, 16 g of clove, 32 g of honeysuckle flower, 8 g of citron, 16 g of astragalus membranaceus, 2 g of houttuynia cordata, 16 g of turmeric, 16 g of phyllanthus emblica, 16 g of rehmannia glutinosa, and 2 g of roselle. After washing and drying, use a pulverizer to break them into powders, and add 10 times the weight of pure water according to the weight of the traditional Chinese medicine composition; add the cultured complex probiotics according to 35% of the weight of the pure water, wherein the complex probiotics are composed of eurotium cristatum, bacillus coagulans, and yeast with a mass ratio of 4:2:1. Adjust the pH to be between 5.6 and 6.6, control the dissolved oxygen at about 9 ppm, ferment at 30 °C for 7 days to obtain the primary fermentation product; after repeating the above operation again, obtain the secondary fermentation product; add 10 times the weight of pure water according to the weight of the above traditional Chinese medicine combined fermentation product, extract with gentle boiling for 2 h, let it stand for 4 h, then take the supernatant and filter it, and concentrate it to a solution with a mass fraction of about 4 times the amount of traditional Chinese medicine input under the environment of vacuum negative pressure of -0.07 to -0.1 MPa and 50 to 65 °C to obtain the crude extract of the traditional Chinese medicine combined fermentation; subject the crude extract of the traditional Chinese medicine combined fermentation to processes such as precipitation, centrifugation, decolorization, and desalting to obtain the said traditional Chinese medicine fermentation extract.

[0032] Example 2

[0033] The difference between this example and Example 1 is that:

[0034] The traditional Chinese medicine composition is: 32 g of sophora japonica flower, 32 g of clove, 32 g of honeysuckle flower, 8 g of citron, 16 g of astragalus membranaceus, 8 g of houttuynia cordata, 2 g of turmeric, 8 g of phyllanthus emblica, 4 g of rehmannia glutinosa, and 4 g of roselle.

[0035] Example 3

[0036] The difference between this example and Example 1 is that:

[0037] The traditional Chinese medicine composition is: 32 g of sophora japonica flower, 16 g of clove, 32 g of honeysuckle flower, 8 g of citron, 2 g of astragalus membranaceus, 2 g of houttuynia cordata, 2 g of turmeric, 8 g of phyllanthus emblica, 8 g of rehmannia glutinosa, and 8 g of roselle.

[0038] Example 4

[0039] The difference between this example and Example 1 is that:

[0040] The traditional Chinese medicine composition is: 32 g of sophora japonica flower, 32 g of clove, 32 g of honeysuckle flower, 8 g of citron, 2 g of astragalus membranaceus, 2 g of houttuynia cordata, 2 g of turmeric, 32 g of phyllanthus emblica, 32 g of rehmannia glutinosa, and 16 g of roselle.

[0041] Comparative Example 1

[0042] A traditional Chinese medicine extract is prepared according to the following steps:

[0043] Weigh 32 g of sophora flower buds, 32 g of cloves, 32 g of honeysuckle flowers, 8 g of citrons, 2 g of astragalus membranaceus, 2 g of houttuynia cordata, 2 g of turmeric, 32 g of phyllanthus emblica, 32 g of rehmannia glutinosa, and 16 g of roselle. After washing, put them in a decoction bag, place them in a decoction machine, add 100 times the weight of pure water according to the weight of the traditional Chinese medicine composition, soak for 4 h, then extract with gentle boiling for 1.5 h. After standing for 4 h, take the supernatant and centrifuge it at 10,000 rpm for 10 min to obtain the supernatant. Under the environment of vacuum negative pressure of -0.07 to -0.1 MPa and 50 to 65 °C, rotary evaporate and concentrate it to a solution with a mass fraction of about 4 times the amount of traditional Chinese medicine input to obtain the traditional Chinese medicine extract.

[0044] Comparative Example 2

[0045] A traditional Chinese medicine fermentation combination is prepared according to the following steps:

[0046] Weigh 32 g of sophora flower buds, 32 g of cloves, 32 g of honeysuckle flowers, 8 g of citrons, 2 g of astragalus membranaceus, 2 g of houttuynia cordata, 2 g of turmeric, 32 g of phyllanthus emblica, 32 g of rehmannia glutinosa, and 16 g of roselle. After washing and drying, use a pulverizer to crush them into powder. Add 10 times the weight of pure water according to the weight of the traditional Chinese medicine composition; add the cultured compound probiotics according to 35% of the weight of pure water, where the compound probiotics are composed of eurotium cristatum, bacillus coagulans, and saccharomyces cerevisiae with a mass ratio of 4:2:1. Adjust the pH to be between 5.6 and 6.6, control the dissolved oxygen at about 9 ppm, and ferment at 30 °C for 7 days to obtain the primary traditional Chinese medicine combination fermentation product.

[0047] Comparative Example 3

[0048] A traditional Chinese medicine fermentation extract is prepared according to the following steps:

[0049] On the basis of Comparative Example 2, add 10 times the weight of pure water according to the weight of the obtained primary traditional Chinese medicine combination fermentation product, extract with gentle boiling for 2 h, stand for 4 h, then filter the supernatant. Under the environment of vacuum negative pressure of -0.07 to -0.1 MPa and 50 to 65 °C, concentrate it to a solution with a mass fraction of about 4 times the amount of traditional Chinese medicine input to obtain the primary traditional Chinese medicine combination fermentation extract.

[0050] Comparative Example 4

[0051] The difference between this example and Example 4 is that:

[0052] The traditional Chinese medicine composition is: 32 g of honeysuckle flowers, 8 g of citrons, 2 g of astragalus membranaceus, 2 g of houttuynia cordata, 32 g of phyllanthus emblica, 32 g of rehmannia glutinosa, and 16 g of roselle.

[0053] Comparative Example 5

[0054] The difference between this example and Example 4 is that:

[0055] The traditional Chinese medicine composition is: 32 g of sophora flower buds, 32 g of cloves, 2 g of houttuynia, 2 g of turmeric, 32 g of phyllanthus emblica, 32 g of rehmannia root.

[0056] Comparative Example 6

[0057] The difference between this example and Example 4 is that:

[0058] The traditional Chinese medicine composition is: 32 g of sophora flower buds, 32 g of cloves, 32 g of honeysuckle, 8 g of citron, 2 g of astragalus membranaceus, 2 g of turmeric, 16 g of roselle.

[0059] Comparative Example 7

[0060] The difference between this example and Example 4 is that:

[0061] The compound probiotics are composed of Eurotium cristatum and Saccharomyces cerevisiae with a mass ratio of 4:1

[0062] Comparative Example 8

[0063] The difference between this example and Example 4 is that:

[0064] The compound probiotics are composed of Eurotium cristatum and Bacillus coagulans with a mass ratio of 2:1

[0065] Comparative Example 9

[0066] The difference between this example and Example 4 is that:

[0067] The compound probiotics are composed of Bacillus coagulans and Saccharomyces cerevisiae with a mass ratio of 2:1

[0068] Comparative Example 10

[0069] The difference between this example and Example 4 is that:

[0070] The compound probiotics are composed of Eurotium cristatum, Bacillus coagulans and Saccharomyces cerevisiae with a mass ratio of 1:2:4.

[0071] Comparative Example 11

[0072] The difference between this example and Example 4 is that:

[0073] The compound probiotics are composed of Eurotium cristatum, Bacillus coagulans and Saccharomyces cerevisiae with a mass ratio of 1:1:1.

[0074] Comparative Example 12

[0075] The difference between this example and Example 4 is that:

[0076] The compound probiotics are composed of Eurotium cristatum, Bacillus coagulans and Saccharomyces cerevisiae in a mass ratio of 5:1:5.

[0077] Test Example 1

[0078] Irritation evaluation

[0079] Test samples: The Chinese medicine fermentation products prepared in Examples 1-4 and Comparative Examples 1-3 were added with excipients to prepare different essence samples. The formulations of the essence samples are shown in Table 1.

[0080] Table 1 Formulation table of essence samples

[0081]

[0082] Preparation method: Prepared according to the following steps:

[0083] (1) Heat phase A to 70 °C and dissolve completely;

[0084] (2) Cool down to 45 °C, add phase B, and stir evenly;

[0085] (3) Add phase C, dissolve, and stir evenly to obtain the essence sample.

[0086] Test method: The erythrocyte hemolysis experiment is one of the alternative methods for the rabbit eye irritation experiment. The basic principle is to evaluate the damage of chemicals to eye tissue cells by measuring the hemoglobin elution amount and denaturation degree. In this test example, according to the erythrocyte hemolysis test method of the European Alternative Method Validation Center (ECVAM.ECVAM DB-ALM: INVITOX Protoco, Redbloodcells test system.INVITOX No.37.Ispra.Italy: ECVAM, 1994.), the grading standard is shown in Table 2, where HD50 is the sample concentration when 50% of the red blood cells are hemolyzed, DI is the protein denaturation index, and L / D is the ratio of HD50 to DI. The essence samples prepared in Examples 1-4 and Comparative Examples 1-3 were taken for the erythrocyte hemolysis experiment, and the experimental results are shown in Table 3.

[0087] Table 2 Irritation grading

[0088]

[0089]

[0090] As can be seen from Table 3, the anti-aging essence samples prepared from the samples of Examples 1-4 were all classified as non-irritating. The HD50 of the anti-aging essence prepared from the samples in Examples 1-4 was higher than 76129 mg / L, with the highest being 79583 mg / L, and the DI was lower than 0.37%, with the lowest being 0.28%, all being non-irritating. In contrast, Comparative Example 1 only went through the process of traditional Chinese medicine extraction and did not undergo traditional Chinese medicine fermentation, resulting in a lower sample concentration when 50% of red blood cells were hemolyzed, and irritation would occur at a relatively low concentration. The HD50 of the anti-aging essence prepared by the combined fermentation (extract) of traditional Chinese medicine in Comparative Examples 2-3 was lower than 69532 mg / L, and the DI was higher than 0.624%, with the highest being 0.68%, indicating that the samples prepared in Comparative Examples 2-3 were also non-irritating, but the concentration at which irritation occurred was relatively lower than that of the examples. This shows that fermenting the traditional Chinese medicine composition with compound probiotics can significantly reduce the toxic and side effects and irritation of the traditional Chinese medicine extract; and the HD50 of the anti-aging essence prepared from the combined fermented product of traditional Chinese medicine in Example 4 was higher than that of Comparative Example 3, and the DI was lower than that of Comparative Example 3, indicating that multiple fermentations can better achieve the purpose of reducing the toxic and side effects and irritation of the traditional Chinese medicine extract compared to single fermentation and direct extraction.

[0091] Irritation results of different samples

[0092] Sample Name <![CDATA[HD 50 (mg / L)]]> DI(%) L / D Evaluation Example 1 76129 0.37 97 Non-irritating Example 2 76524 0.33 >100 Non-irritating Example 3 78321 0.30 >100 Non-irritating Example 4 79583 0.28 >100 Non-irritating Comparative Example 1 52381 0.76 >100 Non-irritating Comparative Example 2 69532 0.68 >100 Non-irritating Comparative Example 3 65856 0.62 >100 Non-irritating

[0093] Test Example 2

[0094] DPPH free radical scavenging ability test

[0095] The ability to scavenge DPPH free radicals can, to a certain extent, reflect the antioxidant ability of the sample. The greater the DPPH free radical scavenging rate, the stronger the antioxidant ability and the stronger the anti-aging ability. Therefore, the anti-aging effect of the sample can be judged by detecting the DPPH free radical scavenging rate.

[0096] Test samples: The samples prepared in Examples 1-4 and Comparative Examples 1-12 were tested.

[0097] Test method: Add 2 mL of DPPH test solution with a mass concentration of 47.7 mg / L and a certain amount of the test sample to a test tube, and make up the total volume to 3 mL with pure water. Shake well, and after reacting in the dark for 30 min, place the mixture in a 1-cm cuvette to detect the absorbance at a wavelength of 517 nm, denoted as As; add 2 mL of pure water and 0.4 mL of the test sample to a test tube, make up the total volume to 3 mL with pure water, and detect its absorbance at a wavelength of 517 nm, denoted as Ar; add 2 mL of DPPH test solution and 1 mL of pure water to a test tube, and detect its absorbance at a wavelength of 517 nm, denoted as Ao.

[0098] The calculation formula for the DPPH free radical scavenging rate (Y, %) is as shown in Equation (1). The DPPH half-scavenging rate (μg / mL) is the concentration of the sample when the DPPH free radical scavenging rate reaches 50%. The lower the DPPH half-scavenging rate, the stronger the ability of the sample to scavenge DPPH free radicals and the better the antioxidant effect.

[0099] Y = [1 - (As - Ar) / Ao] × 100% (1)

[0100] The DPPH half-scavenging rates of the VC standard solution, the samples prepared in Examples 1 - 4, and Comparative Examples 1 - 12 were measured successively according to the above steps, and the results are shown in Table 4.

[0101] Table 4 Test of DPPH Free Radical Scavenging Ability of Different Samples

[0102]

[0103]

[0104] The lower the DPPH half-scavenging rate, the stronger the ability of the sample to scavenge DPPH free radicals and the better the antioxidant effect. As can be seen from Table 4, the DPPH half-scavenging rates of the samples in Examples 1 - 4 are all lower than 16.87 μg / mL, and the lowest is 11.63 μg / mL. In contrast, the DPPH half-scavenging rates of the traditional Chinese medicine fermented products prepared in Comparative Examples 1 - 12 are all higher, all higher than 29.46 μg / mL, and the highest is 51.37 μg / mL. The antioxidant ability of the samples prepared in Examples 1 - 4 is significantly better than that of Comparative Examples 1 - 12. Compared with Example 4, the DPPH half-scavenging rates of Comparative Examples 1 - 3 are about 3.5 - 5 times that of Example 4. This result shows that the preparation method provided by the present invention can significantly improve the antioxidant ability of the traditional Chinese medicine composition. Compared with Example 4, the DPPH half-scavenging rates of Comparative Examples 4 - 6 are about 2.5 - 3 times that of Example 4. This result shows that the various traditional Chinese medicine components in the traditional Chinese medicine fermented extract of the present invention have synergistic effects, interact with each other, and jointly achieve excellent antioxidant effects. Compared with Example 4, the DPPH half-scavenging rates of Comparative Examples 7 - 12 are about 2.5 - 4 times that of Example 4. This result shows that the composite probiotic composed of Eurotium cristatum, Bacillus coagulans, and Saccharomyces cerevisiae with a mass ratio of 4:2:1 used in the traditional Chinese medicine fermented extract of the present invention is a necessary technical means to achieve the antioxidant performance of the traditional Chinese medicine fermented extract.

[0105] Test Example 3

[0106] Effect of the Traditional Chinese Medicine Fermented Extract of the Present Invention on the Viability of Human Epidermal Fibroblasts Irradiated by UV

[0107] Test samples: Samples prepared in Examples 1 - 4 and Comparative Examples 1 - 12.

[0108] Test method:

[0109] Grouping: Each sample was mixed with the culture medium to prepare a test solution, which was the experimental group; based on the culture medium and irradiated with 40 mJ of ultraviolet light, it was set as the model group; the culture medium without light was used as the blank group.

[0110] Test method: Human epidermal fibroblasts in the logarithmic growth phase were taken and inoculated into a 96-well plate at a density of 1×10 4 cells / mL. The samples prepared in Examples 1-4 and Comparative Examples 1-12 were added to each group; the blank group and the model group were added with an equal amount of culture medium. After 24 h of administration, it was irradiated with 40 mJ of ultraviolet light; 24 h after the irradiation ended, the cell viability was detected by CCK8 and the relative content of MMP-1 was detected using a kit (MMP-1, Biotrak Amersham Pharmacia Biotech, United Kingdom; RPN 2610).

[0111] Cell viability (%) = (blank group - experimental group) / blank group × 100;

[0112] MMP-1 inhibition rate (%) = (model group - experimental group) / model group × 100.

[0113] The results of cell viability were as Figure 1 shown. The cell viability of Examples 1-4 was significantly higher than that of the model group, with a significant difference (P < 0.01). The cell viability of Examples 1-4 was all above 60%, and the highest cell viability could reach above 90%. Among them, there was no significant difference between Examples 3-4 and the blank control group. Compared with Example 4, the cell viability of Comparative Examples 1-3 was between 30% and 40%, which was significantly lower than that of Example 4. This result shows that the preparation method provided by the present invention can significantly improve the viability of human fibroblasts treated with UV radiation by the traditional Chinese medicine composition. Compared with Example 4, the cell viability of Comparative Examples 4-6 was between 40% and 60%, which was significantly lower than that of Example 4. This result shows that each traditional Chinese medicine component in the traditional Chinese medicine fermentation extract of the present invention has a synergistic effect, interacts with each other, and jointly enhances the effect of improving the viability of human fibroblasts treated with UV radiation. Compared with Example 4, the cell viability of Comparative Examples 7-12 was between 35% and 60%, which was significantly lower than that of Example 4. This result shows that the composite probiotic composed of Eurotium cristatum, Bacillus coagulans and Saccharomyces cerevisiae with a mass ratio of 4:2:1 used in the traditional Chinese medicine fermentation extract of the present invention is a necessary technical means to achieve the effect of enhancing the viability of human fibroblasts treated with UV radiation by the traditional Chinese medicine fermentation extract.

[0114] The results of the MMP-1 inhibition rate were as Figure 2As shown, the MMP-1 inhibition rates of Examples 1-4 were significantly higher than those of the model group, with significant differences (P<0.01). The MMP-1 inhibition rates of Examples 1-4 were all higher than 60%, and the cell viability could reach up to 90%. Compared with Example 4, the MMP-1 inhibition rates of Comparative Examples 1-3 were between 30-40%, significantly lower than those of Example 4, and this result shows that the preparation method provided by the present invention can significantly improve the ability of the Chinese medicine composition to inhibit the activity of matrix metalloproteinases. Compared with Example 4, the MMP-1 inhibition rates of Comparative Examples 4-6 were between 50-60%, significantly lower than those of Example 4, and this result shows that the various Chinese medicine components in the Chinese medicine fermentation extract of the present invention synergize and interact with each other to jointly enhance the ability to inhibit the activity of matrix metalloproteinases. Compared with Example 4, the cell viability of Comparative Examples 7-12 is between 35-60%, which is significantly lower than that of Example 4. This result shows that the composite probiotic composed of Aspergillus niger, Bacillus coagulans and yeast in a mass ratio of 4:2:1 used in the traditional Chinese medicine fermentation extract of the present invention is a necessary technical means to achieve the effect of the traditional Chinese medicine fermentation extract enhancing the ability to inhibit the activity of matrix metalloproteinases.

[0115] Test Example 4

[0116] Evaluation of anti-aging effects in mice

[0117] Malondialdehyde (MDA) is a metabolite produced under oxidative stress. The degree of intracellular oxidative stress can be verified by measuring the intracellular MDA content.

[0118] Superoxide dismutase (SOD) is an important antioxidant enzyme that protects cells from oxidative stress damage. The determination of SOD content is of great significance for understanding the antioxidant capacity of cells and the degree of oxidative stress.

[0119] Catalase (CAT) is an antioxidant enzyme that is ubiquitous in almost all organisms. Its main function is to catalyze the decomposition of hydrogen peroxide into water and oxygen, remove hydrogen peroxide from the body, and thus protect cells from the toxicity of H2O2. It is one of the key enzymes in the biological defense system.

[0120] Glutathione peroxidase (GSH-Px) is an important peroxide decomposing enzyme widely present in the body. It can reduce toxic peroxides into non-toxic hydroxyl compounds, thereby protecting the structure and function of cell membranes from interference and damage by peroxides.

[0121] Test samples: the Chinese medicinal fermentation extracts prepared in Examples 1-4.

[0122] Test method: After 60 male KM mice were adaptively fed for 1 week, they were randomly divided into 6 groups (10 mice in each group): normal group, model group, and Example 1-4 groups. Except for the normal group which was subcutaneously injected with an equal amount of normal saline, the mice in each experimental group were subcutaneously injected with 5% D-galactose (1000 mg / kg), 0.2 mL per day. Each example and 2% traditional Chinese medicine fermented extract dilution was intragastrically administered 0.2 mL per day, and the normal group and the model group were intragastrically administered an equal volume of distilled water per day. After the experiment, the content of MDA in the mouse serum and the relative activities of SOD, CAT, and GSH-Px were measured.

[0123] The results are as Figure 3 shown. Compared with the model group, the content of MDA in the sera of the mice in Examples 1-4 was significantly decreased, and the relative activities of SOD, CAT, and GSH-Px were all significantly increased, with a significant difference (P<0.01). This result indicates that the traditional Chinese medicine fermented extract provided by the present invention can significantly increase the enzyme activities of SOD, CAT, and GSH-Px in the mouse serum, reduce the MDA activity, and has a significant improvement effect on the aging of mice.

[0124] Test Example 5

[0125] Study on the anti-aging effect of the traditional Chinese medicine fermented extract of the present invention on human skin

[0126] The loss of skin elasticity, the increase in wrinkles, and the roughness of the skin are the most obvious characteristics of skin aging. Therefore, the anti-aging effect of the sample can be evaluated by measuring the changes in skin elasticity, the number of wrinkles, and the smoothness.

[0127] Test samples: Essence samples containing the traditional Chinese medicine fermented extracts prepared in Examples 1-4 were prepared according to the method described in Test Example 1;

[0128] Test method: 70 female volunteers aged 25-40, healthy, with normal skin and no history of cosmetics allergy were selected and evenly divided into 7 groups. Each group of volunteers used the essence samples containing the samples of Examples 1-4 for 30 consecutive days, and a facial image analyzer (Visia), a skin elasticity tester (MPA580), and a skin moisture tester (ProbeCorneometer CM825) were used to track and detect the changes in the facial skin of the volunteers, and the anti-aging effects of the samples were evaluated using skin elasticity, wrinkles, and smoothness as indicators.

[0129] The results are as Figure 4 shown. The main manifestations of human skin aging are the appearance of skin relaxation, wrinkles, roughness, dryness, etc. Figure 4It can be seen that after the volunteers used the samples added with the Chinese medicine fermentation extract of the present invention, the skin elasticity increased by more than 4.5% on average after 30 days, with a maximum of 10.7%, the wrinkles decreased by more than 3.6% on average, with a maximum of 8.5%, the skin smoothness increased by more than 14.5% on average, with a maximum of 20.7%, and the skin aging conditions were significantly improved, indicating that the cosmetics prepared from the Chinese medicine fermentation extract of the present invention have excellent anti-aging effects.

[0130] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A traditional Chinese medicine fermentation extract, which is prepared by co-fermenting a traditional Chinese medicine composition and compound probiotics; the traditional Chinese medicine composition is composed of the following raw materials in parts by weight: 0 - 32 parts of sophora flower buds, 0 - 32 parts of cloves, 0 - 32 parts of honeysuckle, 0 - 32 parts of citrons, 0 - 32 parts of astragalus membranaceus, 0 - 32 parts of houttuynia cordata, 0 - 32 parts of turmeric, 0 - 32 parts of phyllanthus emblica, 0 - 32 parts of rehmannia glutinosa, 0 - 32 parts of roselle; The compound probiotics are one or more of yeast, bacillus coagulans, fungi, and lactobacillus.

2. The fermented and extracted traditional Chinese medicine liquid according to claim 1, wherein, The traditional Chinese medicine composition is composed of the following raw materials in parts by weight: 32 parts of sophora flower buds, 32 parts of cloves, 32 parts of honeysuckle, 8 parts of citrons, 16 parts of astragalus membranaceus, 8 parts of houttuynia cordata, 2 parts of turmeric, 8 parts of phyllanthus emblica, 4 parts of rehmannia glutinosa, 4 parts of roselle.

3. The fermented extract of traditional Chinese medicine according to claim 1, wherein The traditional Chinese medicine composition is composed of the following raw materials in parts by weight: 32 parts of sophora flower buds, 16 parts of cloves, 32 parts of honeysuckle, 8 parts of citrons, 2 parts of astragalus membranaceus, 2 parts of houttuynia cordata, 2 parts of turmeric, 8 parts of phyllanthus emblica, 8 parts of rehmannia glutinosa, 8 parts of roselle.

4. A traditional Chinese medicine fermentation extract according to claim 1, characterized in that, The traditional Chinese medicine composition is composed of the following raw materials in parts by weight: 32 parts of sophora flower buds, 32 parts of cloves, 32 parts of honeysuckle, 8 parts of citrons, 2 parts of astragalus membranaceus, 2 parts of houttuynia cordata, 2 parts of turmeric, 32 parts of phyllanthus emblica, 32 parts of rehmannia glutinosa, 16 parts of roselle.

5. The extract of medium-fermented medicine according to claim 1, characterized in that, The compound probiotics are composed of eurotium cristatum: bacillus coagulans: yeast with a mass ratio of 4 - 2:2 - 0.5:1 - 0.

1.

6. The fermented and extracted traditional Chinese medicine liquid according to claim 1, characterized in that The compound probiotics are composed of eurotium cristatum: bacillus coagulans: yeast with a mass ratio of 4:2:

1.

7. The preparation method of a traditional Chinese medicine fermentation extract according to claim 1, wherein, The traditional Chinese medicine fermentation extract is prepared according to the following steps: (1) Weigh each traditional Chinese medicine raw material according to the above parts by weight, wash, dry and crush, then transfer it to a flask, add 5 - 20 times the weight of pure water, add 5 - 35% of the compound probiotics by volume fraction of pure water in a super clean bench, and carry out a fermentation reaction at 28 - 42 °C for 2 - 10 days to obtain a traditional Chinese medicine ferment; This step is repeated 1 - 3 times; (2) Place the traditional Chinese medicine combined ferment in a decocting machine, add 10 - 20 times pure water according to the weight of the traditional Chinese medicine composition, decoct at a gentle boil for 1.2 - 2 h, then let it stand for 2 - 4 h, take the supernatant and filter, and concentrate it to about 4 - 6 times the mass fraction of the traditional Chinese medicine feeding amount under the environment of vacuum negative pressure of -0.07 ~ -0.1 MPa and 50 - 60 °C to obtain the crude extract of traditional Chinese medicine fermentation; (3) The crude extract of traditional Chinese medicine fermentation is subjected to precipitation, centrifugation, decolorization, and desalting to obtain the final traditional Chinese medicine fermentation extract.

8. The application of a traditional Chinese medicine fermentation extract according to claim 1, wherein, The traditional Chinese medicine fermentation extract is used for preparing anti - aging cosmetics or functional foods; The traditional Chinese medicine fermentation extract is incorporated into anti - aging cosmetics or functional foods at a ratio of 0.1% - 50% by weight.

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