Preparation method and application of ganoderma lucidum and kombucha fermented black tea filtrate

Abstract

The invention discloses a preparation method and application of ganoderma lucidum and kombucha fermented black tea filtrate. According to the preparation method of the ganoderma lucidum and kombucha fermented black tea filtrate, ganoderma lucidum and kombucha are mixed and fermented according to the mass ratio of 1: (0.5-2.5) to obtain fermentation suspension containing hyphae, the fermentation temperature is 25-30 DEG C, and the fermentation time is 100-150 h. And after fermentation, mixing the tea leaves subjected to fixation, moisture regaining, rolling and sterilization with the fermentation suspension, and fermenting again. The temperature of the secondary fermentation is 23-30 DEG C, the time of the secondary fermentation is 5-48 hours, and after the secondary fermentation, the ganoderma lucidum kombucha fermented black tea filtrate is obtained through refining processes of filtering, discharging, sterilizing and the like. The fermented tea provided by the invention is obviously improved in the aspects of polysaccharide content, tea polyphenol content, protein and peptide quantity and the like, has a certain oil control effect, is obviously improved in antioxidant activity, and can be used as a cosmetic raw material to be applied to skin care and washing care products.

Description

Technical Field

[0001] The invention belongs to the technical field of washing, cosmetics and skin care products, and particularly relates to a preparation method and application of black tea filtrate fermented by ganoderma lucidum black tea fungus. Background Art

[0002] As people's living standards improve, they pay more and more attention to hair and skin care. Having healthy hair and smooth and delicate skin can not only improve personal image, but also reflect people's pursuit of healthy life. However, many factors in modern life, such as environmental pollution, bad eating habits, stress, etc., often lead to problems such as split ends, brittleness, and easy breakage of hair, and the skin is also prone to dryness, sensitivity, aging, etc.

[0003] Black tea is rich in a variety of beneficial ingredients, such as tea polyphenols and theaflavins, which have antioxidant and anti-inflammatory effects. These ingredients not only protect the skin, reduce free radical damage to the skin, and delay skin aging, but also nourish the hair and improve its texture and luster. In addition, the nutrients in black tea can also enhance the toughness of hair and reduce the split ends and fragility of hair.

[0004] Microorganisms such as yeast, acetic acid bacteria and lactic acid bacteria in kombucha can produce a variety of beneficial substances during the fermentation process, and these bacteria are naturally symbiotic microorganisms with tea leaves during fermentation, which is conducive to the dissolution and transformation of active substances in kombucha. Literature indicates that kombucha fungus liquid contains more than 50 ingredients that are beneficial to the human body, mainly gluconic acid, acetic acid, lactic acid, folic acid, fructose, glucose, amino acids, proteins, trace elements, tea polyphenols, vitamins, etc. There is a mutually beneficial mixed effect between kombucha fungi. For example, in the initial stage of fermentation, acetic acid bacteria cannot directly use sucrose. The presence of specific enzymes in yeast can decompose sucrose into glucose and fructose, and acetic acid bacteria can continue to oxidize it into substances such as gluconic acid and acetic acid. These metabolites produced by acetic acid further protect the growth of yeast.

[0005] Ganoderma lucidum can produce effective substances such as Ganoderma polysaccharides and Ganoderma triterpenes, which have multiple biological activities such as enhancing immunity, anti-oxidation, and anti-inflammation. Literature studies have shown that the bacterial growth and EPS synthesis of Ganoderma lucidum belong to partially coupled metabolism, and the addition of black tea to the culture medium is conducive to the synthesis of EPS in Ganoderma lucidum. In addition, the growth temperature of Ganoderma lucidum and black tea fungus is similar, 28-30℃, so the mixed fermentation of black tea fungus and Ganoderma lucidum has a certain theoretical basis. Mixing Ganoderma lucidum with black tea and black tea fungus is expected to further enhance the nutritional value and skin care effects of the fermentation products, and provide care for hair and skin.

[0006] Patent publication number CN 115105452 A discloses a black tea fermentation filtrate and its preparation method and application, which uses black tea extract as a substrate and uses yeast, lactic acid bacteria, and acetic acid bacteria for two fermentations, and has moisturizing, anti-aging, anti-inflammatory, and antioxidant effects. This patent uses black tea extract as a substrate for fermentation, which requires special equipment for extraction and increases the process steps, and fails to give full play to the cell wall breaking ability of the strain.

[0007] Patent publication number CN 116334146 A discloses a black tea double yeast fermentation product, cosmetics, preparation method and application thereof, which have antioxidant, anti-aging and brightening effects.

[0008] However, there are relatively few studies on the mixed fermentation of Ganoderma lucidum, black tea and black tea, and related products and technologies still need to be further developed and improved. The addition of black tea is conducive to the production of Ganoderma lucidum polysaccharides, and the addition of Ganoderma lucidum and black tea is conducive to the dissolution and conversion of active substances in black tea leaves; therefore, exploring an effective mixed fermentation method to improve the efficacy of black tea fermentation filtrate is the research focus in this field. Summary of the invention

[0009] In view of the deficiencies in the prior art and the lack of research, the purpose of the present invention is to provide a method for preparing a black tea filtrate fermented by Ganoderma lucidum and kombucha, wherein the filtrate is prepared by adding black tea leaves to Ganoderma lucidum and kombucha as strains for mixed bidirectional fermentation, so that the filtrate is rich in tea polyphenols, Ganoderma lucidum polysaccharides, proteins and peptides, etc., and the skin care effects of anti-oxidation, anti-inflammatory and anti-aging are achieved with natural and safe ingredients.

[0010] Another object of the present invention is to provide an application of the above-mentioned Ganoderma lucidum kombucha fermented black tea filtrate, which can be used as a cosmetic raw material in skin care and washing products.

[0011] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0012] First, the present invention provides a method for preparing a black tea filtrate fermented by Ganoderma lucidum black tea fungus, comprising the following steps:

[0013] 1) preparing a fermentation suspension: mixing Ganoderma lucidum and Kombucha in a mass ratio of 1:(0.5-2.5), and fermenting at 25-30° C. for 100-150 hours to obtain a fermentation suspension containing mycelium;

[0014] 2) Tea pretreatment: the tea leaves are subjected to withering, moisture regaining, rolling and sterilization treatments;

[0015] 3) Fermenting again: mixing the pretreated tea leaves with the fermentation suspension, and fermenting again at 23-30° C. for 5-48 hours;

[0016] 4) After the fermentation is completed, the ganoderma lucidum kombucha fermented black tea filtrate is obtained through purification processes such as filtration and sterilization.

[0017] Preferably, the Ganoderma lucidum is mycelium rather than fruiting body, including any one of red Ganoderma lucidum, antler Ganoderma lucidum, purple Ganoderma lucidum and green Ganoderma lucidum; preferably red Ganoderma lucidum;

[0018] Preferably, the kombucha fungus comprises one or a combination of at least two of yeast, acetobacter, and lactic acid bacteria.

[0019] Preferably, the yeast comprises at least one of Saccharomyces cerevisiae, Saccharomyces ludwigii, Candida krusei, Candida tropicalis, Brettanomyces odorifera, and Debaryomyces hansenii; preferably Saccharomyces cerevisiae.

[0020] Preferably, the acetobacter includes Acetobacter xylinum, Acetobacter xylinum, Gluconobacter, Acetobacter ketogenes, Acetobacter oxidans, and Acetobacter pasteurianus; preferably Acetobacter xylinum.

[0021] Preferably, the lactic acid bacteria include one of Lactobacillus bulgaricus, Lactobacillus rhamnosus, Lactobacillus acidophilus, Bifidobacterium, and Lactobacillus plantarum; preferably Lactobacillus plantarum.

[0022] Preferably, in the preparation method, the mass ratio of Ganoderma lucidum, yeast, and acetic acid bacteria / lactic acid bacteria is 1:(0.5-0.8):(1-1.5).

[0023] Preferably, the tea leaves are black tea, including Dianhong tea from Yunnan, Chuanhong tea from Sichuan and Ninghong tea from Jiangxi; preferably Dianhong tea leaves.

[0024] Preferably, the tea leaves are mixed with the fermentation suspension at a ratio of 1-3% (w / v) for re-fermentation; preferably 2.5% (w / v).

[0025] Preferably, in the method for preparing the filtrate of black tea fermented by Ganoderma lucidum kombucha, the refining process further comprises at least one of cell disruption, centrifugal filtration, concentration and purification.

[0026] Preferably, the concentration method includes at least one of evaporation concentration method, freeze concentration method and absorption concentration method.

[0027] Preferably, the step of obtaining the Ganoderma lucidum kombucha fermented black tea filtrate further comprises the step of adding a preservative.

[0028] Preferably, the preservative is one or a combination of two of phenoxyethanol, sodium benzoate, benzyl alcohol and phenoxyethanol.

[0029] Secondly, the present invention provides the Ganoderma lucidum kombucha fermented black tea filtrate prepared by the method for preparing the Ganoderma lucidum kombucha fermented black tea filtrate as described above.

[0030] Finally, the present invention provides the use of the black tea filtrate fermented by Ganoderma lucidum kombucha as described above, which is used as a cosmetic raw material in skin care and shampoo products.

[0031] Preferably, the shampoo is made from the following ingredients:

[0032] Deionized water: 64.5-91.3wt%;

[0033] Guar hydroxypropyltrimethylammonium chloride: 0.1-0.5wt%;

[0034] Sodium laureth sulfate: 7-15wt%;

[0035] Cocamidopropyl betaine: 1-3wt%;

[0036] Polydimethylsiloxane / polydimethylsiloxane alcohol: 0.2-2wt%;

[0037] Cocamide MEA / cocamide propyl MEA: 0.1-2wt%;

[0038] Suspension stabilizer: 0.2-3wt%;

[0039] Ganoderma lucidum kombucha fermented black tea filtrate: 0.1-10wt%;

[0040] Citric acid: Add appropriate amount to adjust the pH range to between 5-6.

[0041] Preferably, the skin care product is a skin care essence water, and the skin care essence water is made of the following ingredients:

[0042] Thickener: 0.2-1wt%;

[0043] Water: 86.5-99.59%;

[0044] Flavor: 0.01-0.5wt%;

[0045] Squalane: 0.1-2%;

[0046] Ganoderma lucidum kombucha fermented black tea filtrate: 0.1-10wt%.

[0047] Compared with the prior art, the present invention has the following beneficial effects:

[0048] 1) The present invention achieves mutually beneficial synergistic fermentation of ganoderma lucidum, black tea and black tea. On the one hand, the microorganism species naturally symbiotically selected for tea pile fermentation are beneficial to the dissolution and transformation of effective substances in black tea; on the other hand, the addition of black tea is beneficial to the production of EPS by ganoderma lucidum, which increases the efficacy of ganoderma lucidum polysaccharides, ganoderma lucidum triterpenes and the like in the black tea filtrate. The mixed two-way fermentation significantly increases the content of active ingredients such as polysaccharides, tea polyphenols, proteins and peptides in the fermented black tea filtrate.

[0049] 2) The filtrate product of the present invention has a tea polyphenol content of ≥2000ppm and a polysaccharide content of ≥10000ppm, and the ingredients are natural and safe; the inventors conducted efficacy tests on the obtained black tea fermentation filtrate, and the fermented black tea filtrate showed excellent antioxidant and anti-aging effects and a certain oil control effect.

[0050] 3) The color of the cosmetic raw material prepared by the present invention is a reddish brown liquid with slight viscosity, which is consistent with the inherent characteristics of being rich in polysaccharides and ganoderma lucidum black tea.

[0051] 4) The black tea filtrate fermented by ganoderma lucidum kombucha prepared by the present invention is applied to shampoo and skin care products and has good moisturizing and oil control effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] Figure 1 The present invention is a process flow chart for preparing the black tea filtrate fermented by Ganoderma lucidum black tea fungus;

[0053] Figure 2 The appearances of the black tea filtrate fermented with ganoderma lucidum black tea fungus obtained by the present invention, the black tea filtrate fermented with ganoderma lucidum black tea fungus without ganoderma lucidum fungus, and the commercially available double-bacteria black tea. DETAILED DESCRIPTION

[0054] The present invention is further described in detail below by specific examples. It should be understood by those skilled in the art that the examples are only used to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0055] The invention discloses a preparation method and application of a black tea filtrate fermented by ganoderma lucidum and kombucha. The preparation method of the black tea filtrate fermented by ganoderma lucidum and kombucha is to obtain a fermentation suspension containing mycelium by mixed fermentation of ganoderma lucidum and kombucha in a mass ratio of 1: (0.5-2.5), the fermentation temperature is 25-30°C, and the fermentation time is 100-150h. After fermentation, the tea leaves that have been sterilized, rehydrated, rolled, and sterilized are mixed with the fermentation suspension and fermented again. The temperature of the second fermentation is 23-30°C, and the time of the second fermentation is 5-48h. After the second fermentation, the black tea filtrate fermented by ganoderma lucidum and kombucha is obtained by refining processes such as filtration, discharging, and sterilization. The fermented tea provided by the invention has significant improvements in polysaccharide content, tea polyphenol content, protein and peptide quantity, etc., has a certain oil control effect and significantly improves antioxidant activity, and can be used as a cosmetic raw material in skin care and washing products.

[0056] The present invention provides a black tea filtrate fermented by ganoderma lucidum black tea fungus and a preparation method thereof. The preparation process is as follows: Figure 1 The method comprises the following steps: obtaining a fermentation suspension containing mycelium by mixing and fermenting Ganoderma lucidum, yeast, and acetic acid bacteria / lactic acid bacteria in a mass ratio of 1: (0.5-0.8): (1-1.5), wherein the fermentation temperature is 25-30° C. and the fermentation time is 100-150 hours;

[0057] After fermentation, the tea leaves that have been sterilized, moistened, rolled, and sterilized are mixed with the fermentation suspension and fermented again;

[0058] The temperature of the second fermentation is 23-30° C., and the time of the second fermentation is 5-48 hours. After the second fermentation, the Ganoderma lucidum kombucha fermented black tea filtrate is obtained through refining processes such as filtration, discharging, and sterilization.

[0059] The fermented tea provided by the present invention has significantly improved polysaccharide content, tea polyphenol content, protein and peptide content, etc., has a certain oil control effect and significantly improved antioxidant activity, and can be used as a cosmetic raw material in skin care and washing products.

[0060] Figure 2 The appearance of the black tea filtrate fermented with Ganoderma lucidum black tea fungus obtained by the present invention, the black tea filtrate fermented with Ganoderma lucidum black tea fungus without Ganoderma lucidum, and the commercially available double-bacteria black tea. The black tea filtrate fermented with Ganoderma lucidum black tea fungus has a more red color due to the addition of red Ganoderma lucidum, and has the characteristics of both black tea and red Ganoderma lucidum.

[0061] The present invention is further described in detail below through specific preferred embodiments combined with efficacy test examples, but the present invention is not limited to the following embodiments.

[0062] The strain number of G. lucidum karst used in the present invention is GDMCC 5.250;

[0063] The yeast Saccharomyces cerevisiae used in the present invention has a strain number of CGMCC 2.1392;

[0064] The Lactobacillus plantarum (L. plantarum) used in the present invention has a strain number of GDCC 1.1797;

[0065] The embodiments are only used to help understand the present invention and should not be regarded as specific limitations of the present invention. Other commercially available strains are also applicable.

[0066] Example 1

[0067] This embodiment provides a black tea filtrate fermented by Ganoderma lucidum kombucha fungus, and the preparation method thereof is as follows:

[0068] 1) Prepare seed liquid culture medium with 20g / L glucose, 25g / L glycerol, 2.5g / L beef extract powder, 5g / L peptone, 1g / L KH2PO4, 0.5g / LMgSO4·7H2O, 0.05g / L vitamin B1, and 1g / L CaCO3. Sterilize at 121℃ for 20min and cool for later use.

[0069] 2) Prepare a mixed fermentation medium with 35 g / L glucose, 30 g / L glycerol, 2.5 g / L beef extract powder, 5 g / L peptone, 1 g / L KH2PO4, 0.5 g / LMgSO4·7H2O, 0.05 g / L vitamin B1, and 1 g / L CaCO3. Sterilize at 121°C for 20 min, and cool for later use.

[0070] 3) Inoculate the red Ganoderma lucidum on a sterilized PDA agar medium slant and activate the slant for 3-5 days. Take 5 activated Ganoderma lucidum blocks with a diameter of 5 mm, add 100 mL of seed liquid culture medium, and culture at 30°C and 150 rpm / min for 3 days to obtain Ganoderma lucidum seed liquid for use.

[0071] 4) Inoculate brewer's yeast at a 2% inoculum into sterilized PDA liquid culture medium, culture at 28°C, 150 rpm / min for 2 generations, each generation for 2-3 days. At the end of the culture, the OD600 value should be ≥0.5, and the brewer's yeast seed solution is obtained for standby use.

[0072] 5) Inoculate Lactobacillus plantarum in sterilized MRS liquid medium at a 2% inoculum amount, culture at 30° C., 150 rpm / min for 2 generations, each generation for 1-2 days. At the end of the culture, the OD600 value should be ≥0.5 to obtain Lactobacillus plantarum seed solution for standby use.

[0073] 6) Inoculate the mixed fermentation medium with the Ganoderma lucidum seed solution, Saccharomyces cerevisiae seed solution and Lactobacillus plantarum seed solution at a ratio of 1:0.6:1.2, and culture at 30°C and 120 rpm / min for 100 h.

[0074] 7) The pretreated black tea leaves were sterilized at 115° C. for 20 min, and the black tea leaves were added to the fermented mixed fermentation medium obtained in step 6) at a ratio of 2.5% (w / v), and the culture was continued at 30° C. and 150 rpm / min for 10 h.

[0075] 8) Filter to remove tea leaves and particles, collect the filtrate, and sterilize at 85°C for 30 min.

[0076] 9) Add 0.5% phenoxyethanol and 0.5% sodium benzoate as preservatives to obtain the cosmetic raw material of the fermented black tea filtrate of Ganoderma lucidum kombucha fungus.

[0077] Example 2

[0078] This embodiment provides a black tea filtrate fermented by Ganoderma lucidum kombucha fungus, and the preparation method thereof is as follows:

[0079] 1) Prepare seed liquid culture medium with 20g / L glucose, 25g / L glycerol, 2.5g / L beef extract powder, 5g / L peptone, 1g / L KH2PO4, 0.5g / LMgSO4·7H2O, 0.05g / L vitamin B1, and 1g / L CaCO3. Sterilize at 121℃ for 20min and cool for later use.

[0080] 2) Prepare a mixed fermentation medium with 35 g / L glucose, 30 g / L glycerol, 2.5 g / L beef extract powder, 5 g / L peptone, 1 g / L KH2PO4, 0.5 g / LMgSO4·7H2O, 0.05 g / L vitamin B1, and 1 g / L CaCO3. Sterilize at 121°C for 20 min, and cool for later use.

[0081] 3) Inoculate the red Ganoderma lucidum on a sterilized PDA agar medium slant and activate the slant for 3-5 days. Take 5 activated Ganoderma lucidum blocks with a diameter of 5 mm, add 100 mL of seed liquid culture medium, and culture at 30°C and 150 rpm / min for 3 days to obtain Ganoderma lucidum seed liquid for use.

[0082] 4) Inoculate brewer's yeast at a 2% inoculum into sterilized PDA liquid culture medium, culture at 28°C, 150 rpm / min for 2 generations, each generation for 2-3 days. At the end of the culture, the OD600 value should be ≥0.5, and the brewer's yeast seed solution is obtained for standby use.

[0083] 5) Inoculate Lactobacillus plantarum in sterilized MRS liquid medium at a 2% inoculum amount, culture at 30° C., 150 rpm / min for 2 generations, each generation for 1-2 days. At the end of the culture, the OD600 value should be ≥0.5 to obtain Lactobacillus plantarum seed solution for standby use.

[0084] 6) Inoculate the Ganoderma lucidum seed solution, Saccharomyces cerevisiae seed solution, and Lactobacillus plantarum seed solution into the mixed fermentation medium at a ratio of 1:0.8:1.5, and culture at 28°C and 120 rpm / min for 120 h.

[0085] 7) The pretreated black tea leaves were sterilized at 115° C. for 20 min, and the black tea leaves were added to the fermented mixed fermentation medium obtained in step 6) at a ratio of 3% (w / v), and the culture was continued at 25° C. and 150 rpm / min for 36 h.

[0086] 8) Filter to remove tea leaves and particles, collect the filtrate, and sterilize at 85°C for 30 min.

[0087] 9) Add 0.5% phenoxyethanol and 0.5% sodium benzoate as preservatives to obtain the cosmetic raw material of the fermented black tea filtrate of Ganoderma lucidum kombucha fungus.

[0088] Example 3

[0089] This embodiment provides a black tea filtrate fermented by Ganoderma lucidum kombucha fungus, and the preparation method thereof is as follows:

[0090] 1) Prepare seed liquid culture medium with 20g / L glucose, 25g / L glycerol, 2.5g / L beef extract powder, 5g / L peptone, 1g / L KH2PO4, 0.5g / LMgSO4·7H2O, 0.05g / L vitamin B1, 1g / L CaCO3. Sterilize at 121℃ for 20min, cool and set aside

[0091] 2) Prepare a mixed fermentation medium with 35 g / L glucose, 30 g / L glycerol, 2.5 g / L beef extract powder, 5 g / L peptone, 1 g / L KH2PO4, 0.5 g / LMgSO4·7H2O, 0.05 g / L vitamin B1, and 1 g / L CaCO3. Sterilize at 121°C for 20 min, and cool for later use.

[0092] 3) Inoculate the red Ganoderma lucidum on the sterilized PDA agar medium slant and activate the slant for 3-5 days. Take 5 activated Ganoderma lucidum blocks with a diameter of 5 mm, put them into 100 mL seed liquid culture medium, and culture them at 30°C and 150 rpm / min for 3 days to obtain Ganoderma lucidum seed liquid for use.

[0093] 4) Inoculate brewer's yeast at a 2% inoculum into sterilized PDA liquid culture medium, culture at 28°C, 150 rpm / min for 2 generations, each generation for 2-3 days. At the end of the culture, the OD600 value should be ≥0.5, and the brewer's yeast seed solution is obtained for standby use.

[0094] 5) Inoculate Lactobacillus plantarum in sterilized MRS liquid medium at a 2% inoculum amount, culture at 30° C., 150 rpm / min for 2 generations, each generation for 1-2 days. At the end of the culture, the OD600 value should be ≥0.5 to obtain Lactobacillus plantarum seed solution for standby use.

[0095] 6) Inoculate the Ganoderma lucidum seed solution, Saccharomyces cerevisiae seed solution, and Lactobacillus plantarum seed solution into the mixed fermentation medium at a ratio of 1:0.5:1.3, and culture at 30°C and 120 rpm / min for 120 h.

[0096] 7) The pretreated black tea leaves were sterilized at 115° C. for 20 min, and the black tea leaves were added to the fermented mixed fermentation medium obtained in step 6) at a ratio of 1% (w / v), and the culture was continued at 28° C. and 150 rpm / min for 16 h.

[0097] 8) Filter to remove tea leaves and particles, collect the filtrate, and sterilize at 85°C for 30 min.

[0098] 9) Add 0.5% phenoxyethanol and 0.5% sodium benzoate as preservatives to obtain the cosmetic raw material of the fermented black tea filtrate of Ganoderma lucidum kombucha fungus.

[0099] Example 4

[0100] This embodiment provides a black tea filtrate fermented by Ganoderma lucidum kombucha fungus, and the preparation method thereof is as follows:

[0101] The only difference between the method and Example 1 is that the amount of black tea leaves added in step (7) is changed to 1% (w / v), and the amounts of other raw materials and the process are the same as those in Example (1).

[0102] Example 5

[0103] This embodiment provides a black tea filtrate fermented by Ganoderma lucidum kombucha fungus, and the preparation method thereof is as follows:

[0104] The only difference between the method and Example 1 is that the amount of black tea leaves added in step (7) is changed to 3% (w / v), and the amounts of other raw materials and the process are the same as those in Example (1).

[0105] Example 6

[0106] This embodiment provides a black tea filtrate fermented by Ganoderma lucidum kombucha fungus, and the preparation method thereof is as follows:

[0107] The only difference between the method and Example 1 is that the amount of black tea leaves added in step (7) is changed to 4% (w / v), and the amounts of other raw materials and the process are the same as those in Example (1).

[0108] Comparative Example 1

[0109] This comparative example provides a fermented black tea filtrate, and the preparation method thereof is as follows:

[0110] The only difference between the method and Example 1 is that the inoculation ratio of Ganoderma lucidum in step (6) is adjusted to 0. The other raw material dosages and processes are the same as those in Example (1).

[0111] Comparative Example 2

[0112] This comparative example provides a black tea filtrate fermented by Ganoderma lucidum kombucha fungus, and the preparation method thereof is as follows:

[0113] The only difference from Example 1 is that the tea leaves in step (7) are replaced by an extract obtained by soaking the tea leaves in water at a ratio of 4:80 for 2 hours, and the amounts of other raw materials and the process are the same as those in Example (1).

[0114] Comparative Example 3

[0115] This comparative example provides a black tea filtrate fermented by Ganoderma lucidum kombucha fungus, and the preparation method thereof is as follows:

[0116] The only difference from Example 1 is that the tea leaves in step (7) are replaced by an extract obtained by soaking the tea leaves in a ratio of 4:80 for 6 hours, and the amounts of other raw materials and the process are the same as those in Example (1).

[0117] Comparative Example 4

[0118] This comparative example provides a black tea filtrate fermented by Ganoderma lucidum kombucha fungus, and the preparation method thereof is as follows:

[0119] The only difference between the method and Example 1 is that tea leaves are added in step (6). The amounts of other raw materials and the process are the same as those in Example (1).

[0120] The following are the contents of the efficacy test examples.

[0121] Efficacy Test Example 1

[0122] This test example tests the tea polyphenol content of the cosmetic raw materials prepared in Examples 1-6 and Comparative Examples 1-4 and the commercially available double-bacteria black tea:

[0123] Principle: Ferrous tartrate colorimetric method - using the phenol oxygen anions of tea polyphenols to complex with metal ions to develop color and determine the content of tea polyphenols.

[0124] Materials and methods are as follows:

[0125] Reagents:

[0126] Ferric tartrate solution: 1g ferrous sulfate, 5g potassium sodium tartrate, dilute to 1L with water.

[0127] pH-7.5 phosphate buffer: 85mL (liquid a) + 15mL (liquid b) mixed;

[0128] Liquid a: 1 / 15 mol / L disodium hydrogen phosphate solution: 11.876 g Na2HPO4·2H2O, diluted to 1 L with water;

[0129] Liquid b: 1 / 15 mol / L potassium dihydrogen phosphate solution: weigh 9.078 g of KH2PO4 that has been dried in an oven at 110°C for 2 h, and dilute to 1 L with water.

[0130] Method steps: (wavelength 540nm, reagent blank as reference)

[0131] Standard solution: Weigh 0.25 g of pure tea polyphenols and dilute to 250 mL with water to obtain 1 mg / mL.

[0132] Draw a standard curve: transfer 0.0mL; 0.5mL; 1.0mL; 1.5mL; 2.0mL respectively into a 25mL brown volumetric flask, add water to 5mL each, then add 5mL of ferric tartrate solution, dilute to the scale with pH-7.5 phosphate buffer, record the absorbance at 540nm, and draw a standard curve.

[0133] Sample determination: dilute 1 mL of sample 5 times, add 4 mL of water, then add 5 mL of ferric tartrate solution, dilute to 400 nm with pH-7.5 phosphate buffer, record the absorbance at 540 nm, and substitute it into the standard curve equation to obtain the content of tea polyphenols in the black tea fermented by Ganoderma lucidum kombucha.

[0134] The test results are shown in Table 1.

[0135] Table 1 Determination results of tea polyphenol content in Examples 1-6 and Comparative Examples 1-4 and commercially available double-bacteria black tea

[0136]

[0137]

[0138] The test results show that the Ganoderma lucidum kombucha fermented black tea filtrate prepared according to the method for preparing the Ganoderma lucidum kombucha fermented black tea filtrate provided by the present invention has the technical advantage of high tea polyphenol content. Comparison of Examples 1-6 with Comparative Examples 2 and 3 shows that Ganoderma lucidum and kombucha contribute to the dissolution of tea polyphenols. Examples 1-6 add Ganoderma lucidum, and some enzymes contained in Ganoderma lucidum break the cell walls of tea leaves, promote the dissolution of substances in tea leaves, and increase the content of tea polyphenols; while in Comparative Examples 2 and 3, the tea leaves have been removed before adding the bacterial suspension, and the extract used, under such conditions, the tea polyphenols are decomposed less by microbial enzymes.

[0139] Efficacy Test Example 2

[0140] This test example tests the polysaccharide content of the cosmetic raw materials prepared in Examples 1-6 and Comparative Examples 1-4:

[0141] Principle: Phenol-sulfuric acid method for determining polysaccharide content - sugar is hydrolyzed into monosaccharides under the action of concentrated sulfuric acid, and rapidly dehydrated into aldehyde derivatives, which are then condensed with phenol to form an orange-yellow compound with stable color. At a wavelength of 490nm and within a certain concentration range, its absorbance is linearly proportional to the polysaccharide content, so its absorbance can be measured by a spectrophotometer, and the polysaccharide content of the sample can be quantitatively determined using a standard curve.

[0142] Materials and methods are as follows:

[0143] Preparation of standard curve: Accurately weigh an appropriate amount of anhydrous glucose dried to constant weight at 105℃, add water to prepare a stock solution of 0.85mg / ml. Accurately pipette 1.0, 2.0, 3.0, 4.0, 5.0ml, respectively, into a 50ml volumetric flask, add water to the scale, and shake well to obtain the glucose dilution. Then accurately measure 2ml of each of the above dilutions into a stoppered test tube, and accurately measure 2ml of water into another stoppered test tube as a blank control. Add 1ml of 5% phenol solution (freshly prepared) to each tube and shake well, quickly add 5.0ml of concentrated sulfuric acid, shake well immediately, place in a 40℃ water bath for 30min, take out and place in an ice water bath for 5min. Use the blank prepared with the above 2.0ml water as a reference, measure the absorbance at a wavelength of 489nm, and draw a standard curve with the concentration (mg / ml) of the glucose dilution as the horizontal axis and the absorbance as the vertical axis.

[0144] Sample detection: Use a pipette to accurately draw 500μL of the sample to be tested, add water to make up the volume to 2ml, and at the same time accurately measure 2ml of water into another stoppered test tube as a blank control. Add 1ml of 5% phenol solution (freshly prepared) and shake well, quickly add 5.0ml of concentrated sulfuric acid, shake well immediately, and place in a 40℃ water bath for 30min, then take out and place in an ice water bath for 5min. Use the blank prepared with the above 2.0ml of water as a reference, measure the absorbance at a wavelength of 489nm, and compare it with the standard curve to obtain the polysaccharide content.

[0145] The test results are shown in Table 2.

[0146] Table 2 Polysaccharide content determination results of Examples 1-6 and Comparative Examples 1-4

[0147] serial number Polysaccharide content (ppm) Example 1 10980 Example 2 11252 Example 3 9756 Example 4 9713 Example 5 10570 Example 6 9520 Comparative Example 1 820 Comparative Example 2 10757 Comparative Example 3 11000 Comparative Example 4 8290

[0148] The test results show that the Ganoderma lucidum black tea fermented black tea filtrate prepared according to the preparation method of the Ganoderma lucidum black tea fermented black tea filtrate provided by the present invention has the technical advantage of high polysaccharide content. By comparing Examples 1-6, Comparative Examples 2-4 and Comparative Example 1, it can be seen that the contribution of polysaccharides mainly comes from the addition of Ganoderma lucidum.

[0149] By comparing Example 1 and Comparative Example 4, it can be seen that the long-term black tea fermentation by microorganisms is not conducive to the accumulation of tea polyphenols and polysaccharides. The reason may be that the tea polyphenols are degraded by the enzymes secreted by the microorganisms during the long-term fermentation. The reason for the reduction in polysaccharide accumulation may be that the polyphenols in black tea inhibit the growth of Ganoderma lucidum, and the premature addition of black tea makes it impossible for Ganoderma lucidum to grow well.

[0150] Efficacy Test Example 3

[0151] This test example performs LC-MSMS peptide identification test on Example 1 and Comparative Example 1:

[0152] Process: The ultrafiltered and desalted black tea filtrate was subjected to LC-MSMS mass spectrometry analysis. The raw file (RawFile) of the mass spectrometry test was retrieved from the database using the software MAXQUANT 1.5.5.1. The database was derived from the UniProt website (https: / / www.uniprot.org / ) to obtain the peptide information of the black tea filtrate.

[0153] Here’s how:

[0154] The black tea filtrate was subjected to ultrafiltration (10 KDa) and desalting by desalting column.

[0155] Liquid A used in the chromatographic separation liquid phase is a 0.1% formic acid aqueous solution, and liquid B is a 0.1% formic acid acetonitrile aqueous solution (acetonitrile is 84%).

[0156] The liquid chromatography column (0.15 mm*150 mm, RP-C18, Column Technology Inc.) was balanced with 95% A solution, and the sample was loaded into Zorbax 300SB-C18 peptide traps (Agilent Technologies) by an automatic sampler and then separated by the liquid chromatography column. The relevant liquid phase gradient was set as follows:

[0157] 0 min - 50 min, linear gradient of solution B from 4% to 50%;

[0158] 50-54 minutes, linear gradient of solution B from 50% to 100%;

[0159] 54 minutes---60 minutes, liquid B was maintained at 100%.

[0160] The enzymatic products were separated by capillary high performance liquid chromatography and analyzed by mass spectrometry using a Q Exactive HF-X mass spectrometer (ThermoFisher). The analysis time was 60 min. Detection method: positive ion. The following method was used for acquisition: 10 fragment spectra (MS2 scan) were acquired after each full scan (full scan).

[0161] The raw files of mass spectrometry test (Raw File) were searched into the corresponding database using the software MaxQuant 1.5.5.1, and finally the protein identification and quantitative analysis results were obtained. The relevant parameters and instructions used for searching the database are shown in Table 3.

[0162] Table 3 Parameters used by MAXQUANT to check the database

[0163]

[0164] The number of 3-7 peptides and 8-25 peptides obtained by testing is shown in Table 4

[0165] Table 4 Statistical results of mass spectrometry identification

[0166] Sample name (3-7 peptides, 8-28 peptides) Number of peptides Example 1 (3-7, 8-25) 100，203 Example 2 (3-7, 8-25) 105，166 Example 3 (3-7, 8-25) 104，135 Example 4 (3-7, 8-25) 96，223 Example 5 (3-7, 8-25) 106，147 Example 6 (3-7, 8-25) 103，126 Comparative Example 1 (3-7, 8-25) 65，100 Comparative Example 2 (3-7, 8-25) 63，79 Comparative Example 3 (3-7, 8-25) 89，95 Comparative Example 4 (3-7, 8-25) 118，16

[0167] The test results show that the fermented black tea filtrate of Ganoderma lucidum Kombucha has more polypeptides. The gene names obtained by searching the database were searched in the NCBI database, and GO functional enrichment analysis was performed. The HYTEA_000119 protein present in the fermented black tea filtrate has a zinc finger binding domain, indicating that the black tea filtrate may have an oil control effect. The molecular function of the HYC85_ series of proteins shows that it has a functional region related to antioxidants, indicating that the fermented black tea filtrate may have good antioxidant properties.

[0168] Efficacy Test Example 4

[0169] This experimental example conducts DPPH-free radical scavenging experiments on Examples 1-6, Comparative Examples 1-4, and commercially available double-bacteria black tea:

[0170] Principle: 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH for short) is a stable long-lived free radical. Its ethanol solution is dark purple and has strong absorption near 517nm. When there is a free radical scavenger, the light absorption of DPPH ethanol solution is weakened due to pairing with its single electron. The degree of fading of DPPH ethanol solution is linearly related to the number of electrons it accepts, which can be used to evaluate the ability of the test sample to scavenge free radicals, that is, the size of the antioxidant activity.

[0171] Materials and methods are as follows:

[0172] Test treatment: Dilute the sample tenfold with water to obtain a multi-level concentration sample. Refer to Table 5, use 10mL test tubes to set up sample tubes (T), sample background (T0), DPPH tubes (C) and solvent background (C0). For each sample tube (T) of each test concentration, three parallel tubes need to be set up, and three parallel tubes need to be set up for the DPPH tube (C). Add 1mL of sample solution of the same concentration to each sample tube (T) and sample background (T0). Add water to all test tubes (T, T0, C, C0) to make up to 3mL and mix well. Add 1mL of DPPH ethanol solution to the sample tube (T) and DPPH tube (C), replace the sample background (T0) and solvent background (C0) with 95% ethanol, shake gently, and let stand at room temperature for 5 minutes. Transfer each reaction solution into a 1cm cuvette and measure the absorbance at 517nm.

[0173] Table 5 Sample addition requirements

[0174] T-sample tube <![CDATA[T0 - Sample background]]> C-DPPH tube <![CDATA[C0 - Solvent Background]]> Sample solution (mL) 1 1 - - Water (mL) 2 2 3 3 DPPH ethanol (mL) 1 - 1 - Parallel times 3 1 3 1

[0175] Calculate the DPPH free radical scavenging rate:

[0176] Clearance (%) = (1-(T-T0) / (C-C0)) × 100%;

[0177] In the formula: T is the absorbance of the sample tube, i.e. the absorbance of the solution after the sample reacts with DPPH; T0 is the background absorbance of the sample; C is the average of the absorbance of the DPPH tube three times, i.e. the absorbance of the DPPH solution when no sample is added; C0 is the background absorbance of the solvent.

[0178] Table 6 DPPH free radical scavenging rate test results of Examples 1-6 and Comparative Examples 1-4 and commercially available double bacteria black tea

[0179] serial number DPPH free radical scavenging rate of 10% diluted solution (%) Example 1 90.39 Example 2 88.73 Example 3 82.85 Example 4 86.13 Example 5 90.05 Example 6 89.79 Comparative Example 1 75.87 Comparative Example 2 70.43 Comparative Example 3 78.35 Comparative Example 4 44.7 Commercially available double bacteria black tea 34.87

[0180] The results of DPPH free radical scavenging rate show that the black tea filtrate fermented by Ganoderma lucidum kombucha provided by the present invention has the technical advantage of strong antioxidant capacity. By comparing Examples 1-6 with Comparative Examples 1-4 and commercially available double-bacteria black tea, it can be concluded that the polysaccharides in the black tea filtrate fermented by Ganoderma lucidum kombucha of the present invention contribute to the antioxidant activity, and the synergistic effect of Ganoderma lucidum polysaccharides and tea polyphenols improves the antioxidant activity of the fermentation filtrate.

[0181] Efficacy Test Example 5

[0182] This test example conducts superoxide anion free radical scavenging test on Examples 1-6 and Comparative Examples 1-4:

[0183] Principle: Pyrogallol auto-oxidation method: Pyrogallol auto-oxidation-spectrophotometry is widely used in the preliminary screening of antioxidants in the food and pharmaceutical industries and the evaluation of the antioxidant function of various active substances in scavenging superoxide anion free radicals O2-·. The pyrogallol auto-oxidation process is a chain reaction that can produce O2-·. The content of its own oxidation product can be detected by a spectrophotometer. When the pH value is less than 9.0, the pyrogallol auto-oxidation rate is positively correlated with the concentration of the generated O2-·. Therefore, the scavenging effect of antioxidants on O2-· in this system can be quantitatively determined by a UV-visible spectrophotometer, and the antioxidant capacity of antioxidants can be indirectly evaluated.

[0184] Materials and methods are as follows:

[0185] (1) Take 5 mL of 0.05 mol / L Tris-HCl buffer (pH 8.2) in a test tube and keep it in a 25℃ water bath for 20 min. Add 4 ml of distilled water and continue to preheat it in a 25℃ water bath for 20 min. Then add 1 ml of 3 mmol / L pyrogallol solution (preheated in a 25℃ water bath for 20 min in advance). After mixing, react at 25℃ water bath for 5 min. Then add 2 ml of 6 mol / L HCl solution to terminate the reaction. Measure the absorbance at 320 nm. Perform three parallel experiments and take the average value, which is recorded as A1 (blank group).

[0186] (2) Place 5 mL of 0.05 mol / L Tris-HCl buffer (pH 8.2) in a test tube and keep it in a 25°C water bath for 20 min. Then add 4 mL of the sample solution to be tested and continue preheating in a 25°C water bath for 20 min. Then add 1 mL of 3 mmol / L pyrogallol solution (preheated in a 25°C water bath for 20 min in advance). Mix well and react in a 25°C water bath for 5 min. Then add 2 mL of 6 mol / L HCl solution to terminate the reaction. Measure the absorbance at 320 nm. Perform three parallel experiments and take the average value, which is recorded as A3 (experimental group).

[0187] (3) Measure the absorbance at 320 nm of the sample solution of the same concentration and record it as A2 (eliminating the influence of sample color).

[0188] Let A = A3-A2, then the removal rate of O2-· by the sample solution can be expressed as:

[0189] SA / %=(A1-A) / A1×100%

[0190] Table 7 Superoxide anion free radical scavenging rate measurement results of Examples 1-6 and Comparative Examples 1-4

[0191]

[0192]

[0193] The results of the superoxide anion free radical scavenging experiment show that the black tea filtrate fermented by Ganoderma lucidum kombucha fungus provided by the present invention has a certain ability to scavenge superoxide anion free radicals.

[0194] Efficacy Test Example 6

[0195] This test example conducts a 5a reductase inhibition test on Example 1 and Comparative Example 1:

[0196] Principle: The anatomy and histology of human skin indicate that the sebaceous gland is a vesicular gland that opens mostly at the top of the hair follicle. Its secretion also increases after the development of the second characteristic, and is significantly affected by male hormones. The sebaceous gland is an important gland attached to the skin and an important part of sebum production. It can secrete sebum, which enters the follicle and is discharged to the surface of the skin through the pores. The sebaceous gland is the target organ of androgen. Testosterone increases the volume of the sebaceous gland and increases its secretion. In particular, the overactivity of 5α-reductase, a catalyst for sebum synthesis in the human body, causes an increase in the secretion of dihydrotestosterone (DHT, a male hormone), which causes the sebaceous gland to increase in dysfunction. 5α-reductase is an enzyme that converts free testosterone into DHT in the human body. Without 5α-reductase, DHT cannot be formed. Therefore, reducing the level of DHT by inhibiting the activity of 5α-reductase can effectively alleviate the excessive secretion of oil by the sebaceous gland, thereby achieving the effect of oil control.

[0197] 5a-reductase is a membrane protease that depends on reduced coenzyme II (NADPH). It is an important male hormone metabolic enzyme in the skin. It can irreversibly convert testosterone (T) into dihydrotestosterone (DHT). DHT is the most active male hormone and can induce excessive oil secretion from sebaceous glands. Reducing DHT levels by inhibiting 5a-reductase activity can effectively relieve excessive oil secretion from sebaceous glands.

[0198] Through the experimental design, the test sample and the blank control group were compared for the 5α-reductase inhibition rate test results. If the inhibition rate of the test sample is better (higher) than that of the blank control group and the statistical difference P value is <0.05, it can be considered that the test sample has a certain oil control effect.

[0199] Test method: Entrusted Yinger Testing Technology Service (Guangdong) Co., Ltd. to conduct the test to observe the oil control effect of the present invention. The test concentration was 200 mg / mL, and the positive control was 2.5 mmol / L of Dutagon solution. The results are summarized as follows.

[0200] Table 8 5a-Reductase inhibition rate measurement results of Example 1 and Comparative Example 1

[0201] Experimental Project Blank control group Example 1 Comparative Example 1 Positive Control 5a-reductase inhibition rate % 0.76 36.14 39.05 78.32 p-value / p<0.05 p<0.05

[0202] The test results show that: the inhibition rate of the positive control group is higher than that of the blank control group and the difference is significant (p<0.05), so the test results are valid; the inhibition rates of Example 1 and Comparative Example 1 are higher than those of the blank control group and the difference is significant (p<0.5), which indicates that the black tea fermentation filtrate itself has a certain oil control effect, and the synergistic fermentation of Ganoderma lucidum has little effect on the oil control effect.

[0203] The following are application examples of shampoo containing black tea filtrate fermented by Ganoderma lucidum kombucha fungus.

[0204] Application Example 1

[0205] A shampoo containing a filtrate of black tea fermented by ganoderma lucidum kombucha, the raw material components of the shampoo comprising:

[0206] Deionized water: 80.2wt%;

[0207] Guar hydroxypropyltrimonium chloride: 0.2wt%;

[0208] Sodium laureth sulfate: 9.1wt%;

[0209] Cocamidopropyl betaine: 3wt%;

[0210] Dimethicone / dimethiconol: 1wt%;

[0211] Cocamide MEA / cocamide propyl MEA: 1.3wt%;

[0212] Suspension stabilizer: 0.2wt%;

[0213] Ganoderma lucidum kombucha fermented black tea filtrate: 5wt%;

[0214] Citric acid: Add appropriate amount to adjust the pH range to between 5-6.

[0215] The preparation method of the shampoo described in this application example is as follows:

[0216] Add water and guar hydroxypropyltrimonium chloride to the reaction vessel and mix. Add sodium laureth sulfate, cocamidopropyl betaine, cocamide MEA and heat to 70-85°C and stir for 30 minutes; cool to 40°C and add suspension stabilizer containing hydrogenated castor oil, polydimethylsiloxane / polydimethylsiloxane alcohol, ganoderma lucidum kombucha fermented black tea filtrate, preservatives, and appropriate amount of citric acid and mix well.

[0217] Application Comparative Example 1

[0218] The shampoo in the comparative example 1 of this application does not contain the fermentation filtrate of Ganoderma lucidum kombucha, and other operations are the same as those in the application example 1.

[0219] The following is part of the efficacy test example.

[0220] Efficacy Test Example 7

[0221] This shampoo was tested on 30 volunteers, including 15 boys and 15 girls, for a two-week use test. After two weeks, sensory evaluation was conducted on the smoothness of dry hair, moisturizing effect, oil control effect, and fluffy effect. Satisfied means a full score of 5 points, and unsatisfied means 0 points.

[0222] Table 8 Sensory evaluation results of shampoo made from black tea filtrate fermented with Ganoderma lucidum kombucha

[0223] Application Example 1 Application Comparative Example 1 P-value Dry hair smoothness 4.29 4.10 P>0.05 Moisturizing effect 4.51 3.76 P<0.05 Oil control effect 4.74 2.36 P<0.05 Overall satisfaction 4.8 3.42 P<0.05

[0224] Application Example 2

[0225] This application example provides an essence water containing a filtrate of black tea fermented by Ganoderma lucidum kombucha, and the raw material components of the essence water include:

[0226] Ammonium acryloyldimethyltaurate / VP copolymer (thickener): 0.6wt%;

[0227] Water: 97.27wt%;

[0228] Fragrance: 0.03wt%;

[0229] Squalane: 0.1wt%;

[0230] Ganoderma lucidum kombucha fermented black tea filtrate: 2wt%.

[0231] The preparation method of the essence water described in this application example is as follows:

[0232] Weigh the thickener and water, mix and stir, and homogenize for 5 minutes. Add the essence, squalane, and the filtrate of the fermented black tea of ​​Ganoderma lucidum kombucha fungus in turn and stir evenly to obtain the essence water.

[0233] Application Comparative Example 2

[0234] The essence water in the comparative application example 2 does not contain the fermentation filtrate of Ganoderma lucidum kombucha, and other operations are the same as those in the application example 2.

[0235] The following is part of the efficacy test example.

[0236] Efficacy Test Example 8

[0237] The test of essence water mainly tests two dimensions: using C+K skin tester to test transepidermal water loss and oil control effects.

[0238] 30 volunteers aged 20 to 40, 15 men and 15 women, were recruited to conduct transepidermal water loss test on the arm and oil control effect test on the forehead. Before using the essence water, the transepidermal water loss of the volunteers was tested using the Tewameter TM Hex probe. Three points were taken for each sample and the average value was A1. The sebum secretion of the volunteers was tested using the Cartridge for the Sebumeter SM815. Three points were taken and the average value was B1. Eight hours after using the essence water, the transepidermal water loss was measured again. Three points were taken for each sample and the average value was A2. The sebum secretion was tested. Three points were taken and the average value was B2. The improvement of transepidermal water loss and oil secretion is summarized as follows.

[0239] Improvement of transepidermal water loss = (A1-A2) / A1;

[0240] Improvement in oil secretion (oil control) = (B1-B2) / B1.

[0241] Table 9 Improvement of transepidermal water loss and oil secretion

[0242] Application Example 2 Application Comparative Example 2 P-value Improvement of transepidermal water loss (%) 7.8％ 0.45％ <0.05 Improvement in oil secretion (%) 16.51％ 2.56％ <0.05

[0243] The test results of shampoo application example 1 and comparative example 1 and essence water application example 2 and comparative example 2 show that the black tea filtrate fermented by Ganoderma lucidum kombucha fungus has good moisturizing effect and oil control effect.

[0244] In summary, the black tea filtrate fermented by Ganoderma lucidum kombucha of the present invention has good moisturizing and oil-controlling effects when applied to shampoo and skin care products.

[0245] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form. Therefore, any simple modification, equivalent change and modification made to the above embodiment according to the technical essence of the present invention without departing from the technical solution of the present invention still fall within the scope of the technical solution of the present invention.

Claims

1. A method for preparing a filtrate of black tea fermented by Ganoderma lucidum kombucha, It is characterized in that The method comprises the steps of: 1) preparing a fermentation suspension: mixing Ganoderma lucidum and Kombucha in a mass ratio of 1:(0.5-2.5), and fermenting at 25-30° C. for 100-150 hours to obtain a fermentation suspension containing mycelium; 2) Tea pretreatment: the tea leaves are subjected to withering, moisture regaining, rolling and sterilization treatments; 3) Fermenting again: mixing the pretreated tea leaves with the fermentation suspension, and fermenting again at 23-30° C. for 5-48 hours; 4) After the fermentation is completed, the ganoderma lucidum kombucha fermented black tea filtrate is obtained through purification processes such as filtration and sterilization.

2. The method for preparing the black tea filtrate fermented by Ganoderma lucidum kombucha according to claim 1, Features: The ganoderma lucidum is the mycelium of ganoderma lucidum instead of the fruiting body, and includes any one of red ganoderma lucidum, deer ganoderma lucidum, purple ganoderma lucidum, and green ganoderma lucidum; preferably red ganoderma lucidum; The kombucha fungus includes one or a combination of at least two of yeast, acetobacter, and lactic acid bacteria; The tea is a kind of black tea, including Dianhong tea from Yunnan, Chuanhong tea from Sichuan and Ninghong tea from Jiangxi; Dianhong tea from places with higher altitudes is preferred; The tea leaves are mixed with the fermentation suspension at a ratio of 1-3% (w / v) and fermented again.

3. The method for preparing the black tea filtrate fermented by Ganoderma lucidum kombucha according to claim 2, Features: The yeast comprises at least one of Saccharomyces cerevisiae, Saccharomyces ludwigii, Candida krusei, Candida tropicalis, Brettanomyces odorifera, and Debaryomyces hansenii; preferably Saccharomyces cerevisiae; The acetic bacteria include Acetobacter xylinum, Acetobacter xylinum, Gluconobacter, Acetobacter ketogenes, Acetobacter oxidans, and Acetobacter pasteurianus; Acetobacter xylinum is preferred; The lactic acid bacteria include one of Lactobacillus bulgaricus, Lactobacillus rhamnosus, Lactobacillus acidophilus, Bifidobacterium, and Lactobacillus plantarum; preferably Lactobacillus plantarum.

4. The method for preparing the black tea filtrate fermented by Ganoderma lucidum kombucha according to claim 2, Features: The mass ratio of the ganoderma lucidum, yeast, and acetic acid bacteria / lactic acid bacteria is 1:(0.5-0.8):(1-1.5).

5. The method for preparing the black tea filtrate fermented by Ganoderma lucidum kombucha according to claim 1, Features: The refining process also includes at least one of cell disruption, centrifugal filtration, concentration, and purification; The concentration method includes at least one of evaporation concentration method, freeze concentration method and absorption concentration method.

6. The method for preparing the black tea filtrate fermented by Ganoderma lucidum kombucha according to claim 1, Features: The method of obtaining the black tea filtrate fermented by Ganoderma lucidum kombucha also includes the step of adding a preservative; the preservative is one or a combination of two of phenoxyethanol, sodium benzoate, benzyl alcohol and phenanthroline.

7. The Ganoderma lucidum kombucha fermented black tea filtrate obtained by the method for preparing the Ganoderma lucidum kombucha fermented black tea filtrate according to any one of claims 1 to 6.

8. The use of the black tea filtrate fermented by Ganoderma lucidum kombucha according to claim 7, Features: As a cosmetic raw material, it is used in skin care, shampoo and other products.

9. The use of the black tea filtrate fermented by Ganoderma lucidum kombucha as claimed in claim 8, Features: The ganoderma lucidum kombucha fermented black tea filtrate is applied to shampoo, and the shampoo is made of the following ingredients: Deionized water: 64.5-91.3wt%; Guar hydroxypropyltrimethylammonium chloride: 0.1-0.5wt%; Sodium laureth sulfate: 7-15wt%; Cocamidopropyl betaine: 1-3wt%; Polydimethylsiloxane / polydimethylsiloxane alcohol: 0.2-2wt%; Cocamide MEA / cocamide propyl MEA: 0.1-2wt%; Suspension stabilizer: 0.2-3wt%; Ganoderma lucidum kombucha fermented black tea filtrate: 0.1-10wt%; Citric acid: Add appropriate amount to adjust the pH range to between 5-6.

10. The use of the black tea filtrate fermented by Ganoderma lucidum kombucha as claimed in claim 8, Features: The ganoderma lucidum kombucha fermented black tea filtrate is used in skin care products, and the skin care products are skin care essence water, and the skin care essence water is made of the following ingredients: Thickener: 0.2-1wt%; Water: 86.5-99.59%; Flavor: 0.01-0.5wt%; Squalane: 0.1-2%; Ganoderma lucidum kombucha fermented black tea filtrate: 0.1-10wt%.

Images