CN111759770B - Traditional Chinese medicine composition bidirectional fermentation mycoplasm extract and preparation method and application thereof - Google Patents
Traditional Chinese medicine composition bidirectional fermentation mycoplasm extract and preparation method and application thereof Download PDFInfo
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Abstract
The invention provides a traditional Chinese medicine composition bidirectional fermentation mycoplasm extractive, a preparation method and application thereof, wherein the preparation method comprises the following steps: (1) Extracting cortex moutan and herba Saussureae Involueratae as raw materials to obtain Chinese medicinal composition extract; (2) Adding the Chinese medicinal extract into the basic culture medium inoculated with and cultured with Ganoderma mycelium, fermenting, and filtering to obtain fermented mycoplasm; (3) Extracting the fermentation mycoplasm to obtain the traditional Chinese medicine composition bidirectional fermentation mycoplasm extract. The traditional Chinese medicine combined bidirectional fermentation mycoplasm extractive obtained by the invention has high polysaccharide and flavone contents, strong inhibition capacity on tyrosinase, strong inhibition capacity on hyaluronidase and strong scavenging capacity on DPPH and hydroxyl free radicals. It can be used as potential cosmetic whitening, anti-aging and anti-allergy raw materials, and is applied to cosmetic products.
Description
Technical Field
The invention belongs to the technical field of fine chemical engineering, particularly relates to a traditional Chinese medicine composition bidirectional fermentation mycoplasm extract, and a preparation method and application thereof, and particularly relates to a traditional Chinese medicine composition bidirectional fermentation mycoplasm extract with whitening, anti-aging and allergy-relieving effects, and a preparation method and application thereof.
Background
Ganoderma is also called Ganoderma lucidum, ganoderma sinense, ganoderma lucidum, and Rumey, and has pharmacological functions of protecting liver, resisting tumor, resisting HIV-1 and HIV-1 protease activity, resisting histamine release, inhibiting angiotensin, regulating immunity, resisting tumor, resisting oxidation, resisting aging, and regulating blood sugar. Cortex moutan is dry root bark of Paeonia suffruticosa Andr.f. of Ranunculaceae, collecting root in autumn, removing fine root and silt, stripping off root bark, sun drying or scraping off coarse bark, removing Cor, and sun drying. The former is called Lian Dan Pi, and the latter is called Shu Dan Pi. Bitter and pungent with slight cold, and entering heart, liver and kidney channels, has effects of clearing heat and cooling blood, promoting blood circulation and removing blood stasis, and can be used for treating heat entering nutrient-blood, warm toxicity and macula, hematemesis and epistaxis, night fever and morning coldness, anhidrosis and bone steaming, amenorrhea and dysmenorrhea, traumatic injury pain, carbuncle and swelling and sore. Snow lotus herb is a folk medicinal plant in alpine regions, is used for dispelling cold and removing dampness, promoting blood circulation and dredging channels, resisting inflammation and relieving pain and the like, and is folk used for treating rheumatic arthritis, gynecological diseases and the like. Pharmacological experiments prove that the snow lotus herb has the effects of resisting cancer, expanding blood vessels, reducing blood pressure, resisting fatigue and the like.
The bidirectional fermentation engineering of medicinal fungus is a composite Chinese medicine production process combining medicinal fungus and Chinese medicinal materials, and has medicinal fungus as fermenting strain, active Chinese medicinal materials as matrix component, and the matrix provides nutrients for the growth of fungus and produces new components through decomposition or synthesis of fungus, so that the fermentation has bidirectional effect. The fermentation product is called medicinal mycoplasm, the mycoplasm not only has the components and effects of medicinal fungi and Chinese medicinal herbs, but also can possibly generate new medicinal substances and effects, and different fungi and different Chinese medicinal herbs are crossed and combined to form almost unlimited fermentation combinations to generate various mycoplasm, so that the mycoplasm obtained by the medicinal fungus bidirectional fermentation engineering can be researched and developed as an innovative Chinese medicament. In recent years, the bidirectional fermentation technology of ganoderma lucidum has made a certain research progress, and researchers culture ganoderma lucidum by ginkgo biloba, figwort root, astragalus root, angelica, codonopsis pilosula powder or water extract thereof, and find that the inoxidizability of fermentation mycoplasm is obviously improved under the condition of proper addition amount.
CN102614238A discloses an application of Ganoderma Ginseng radix medicinal mycoplasm in preparing medicine for treating lung cancer, wherein the Ganoderma Ginseng radix medicinal mycoplasm is prepared by taking Ginseng radix as matrix and Ganoderma as strain, and performing bidirectional solid fermentation, contains various ginsenosides Rg1, rb1, re, rg3, rh1 and the like, and also contains components such as ginseng polysaccharide, ganoderma lucidum polysaccharide and the like, and can be used for preparing medicine for treating lung cancer.
CN103355666A discloses a preparation method of a plant enzyme beverage capable of improving cardiovascular and cerebrovascular metabolism, which comprises soaking fructus Hordei Germinatus, fructus Setariae Germinatus, radix Puerariae, radix Notoginseng, radix Salviae Miltiorrhizae, cortex Eucommiae, rhizoma Chuanxiong, rhizoma corydalis, radix Polygalae, ganoderma, radix Polygoni Multiflori Preparata, rhizoma Polygonati Odorati, radix astragali, pericarpium Citri Reticulatae, flos Sophorae, fructus crataegi, fructus Mori, semen Cassiae, cortex Eucommiae, and herba Gynostemmatis in water, decocting, forcibly circulating the extractive solution under conventional hot reflux, adding sugar and peptone, blending, sterilizing, and filtering to obtain a filtrate as nutrient solution; inoculating probiotic strain to the nutrient solution in a fermentation tank at 30 deg.C, mixing and fermenting in four stages, and quality detecting to obtain beverage.
The invention aims to obtain a bidirectional fermentation mycoplasm extract with whitening, anti-aging and allergy-relieving effects by a bidirectional fermentation technology of lucid ganoderma and a traditional Chinese medicine composition, and develops the application of the bidirectional fermentation mycoplasm extract in the field of cosmetics.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a traditional Chinese medicine composition bidirectional fermentation mycoplasm extract and a preparation method and application thereof, and particularly provides the traditional Chinese medicine composition bidirectional fermentation mycoplasm extract with the effects of whitening, resisting aging and relieving allergy and the preparation method and application thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
in one aspect, the invention provides a preparation method of a bidirectional fermentation mycoplasm extractive of a traditional Chinese medicine composition, which comprises the following steps:
(1) Extracting moutan bark and saussurea involucrate serving as raw materials to obtain a traditional Chinese medicine composition extract;
(2) Adding the Chinese medicinal extract obtained in step (1) into a basic culture medium inoculated with and cultured with ganoderma lucidum mycelia, fermenting, and filtering to obtain fermented mycoplasm;
(3) And (3) extracting the fermentation mycoplasm obtained in the step (2) to obtain the traditional Chinese medicine composition bidirectional fermentation mycoplasm extract.
The invention specifically selects two traditional Chinese medicine components of tree peony bark and snow lotus, the two traditional Chinese medicine components are cooperated, the traditional Chinese medicine composition extract obtained by extracting the traditional Chinese medicine composition extract is used as a fermentation substrate component, lucid ganoderma is used as a fermentation strain, the bidirectional fermentation is carried out to obtain a mycoplasm component, and the mycoplasm component is further extracted to obtain the bidirectional fermentation mycoplasm extract of the traditional Chinese medicine composition. The obtained traditional Chinese medicine combination bidirectional fermentation mycoplasm extractive is superior to the lucid ganoderma fermentation mycoplasm extractive without the traditional Chinese medicine combination extractive and the bidirectional fermentation mycoplasm extractive only added with one of peony and snow lotus in the aspects of polysaccharide and flavone content, the inhibition capacity on tyrosinase, the inhibition capacity on hyaluronidase and the scavenging capacity on DPPH and hydroxyl free radicals. It can be used as potential cosmetic whitening, anti-aging and anti-allergy raw materials, and is applied to cosmetic products.
In the invention, the mass ratio of the moutan bark to the saussurea involucrate in the step (1) is (1-5) 1, for example, 1.
The mass ratio of the moutan bark to the saussurea involucrate is (1-5): 1, the mutual matching relation of the moutan bark and the saussurea involucrate is more obvious and effective, and the moutan bark and the saussurea involucrate are more excellent in the content of polysaccharide and flavone, the inhibition capability on tyrosinase, the inhibition capability on hyaluronidase and the scavenging capability on DPPH and hydroxyl free radicals.
Preferably, the solvent used in the extraction of step (1) comprises water and/or ethanol, preferably an aqueous ethanol solution.
Preferably, the mass concentration of the ethanol aqueous solution is 40-60%, such as 40%, 45%, 50%, 55%, or 60%, and other specific values within the above range can be selected, and are not described herein again.
The mass concentration of the ethanol aqueous solution is also one of key factors influencing the beneficial effects of the final mycoplasm extract, and the specific selection is 40-60%, because the effective components extracted by the ethanol aqueous solution at the concentration are more suitable for being used as matrix components of subsequent bidirectional fermentation, and the effective components are more excellent in the contents of polysaccharide and flavone, the inhibition capability on tyrosinase, the inhibition capability on hyaluronidase and the scavenging capability on DPPH and hydroxyl radicals.
Preferably, the mass ratio of the ethanol aqueous solution to the raw materials is (8-12) 1, for example, 8.
Preferably, the extraction temperature in step (1) is 70-90 ℃, for example, 70 ℃, 75 ℃, 80 ℃, 85 ℃ or 90 ℃, the extraction time is 1-3h, for example, 1h, 1.5h, 2h, 2.5h or 3h, and the extraction frequency is 1-3 times, for example, 1 time, 2 times or 3 times, and other specific points in the above range can be selected, and thus, the description is omitted.
The temperature and the time of the extraction are also one of the key factors influencing the beneficial effects of the final mycoplasm extract, the specific selection is 70-90 ℃, and the time is 1-3h, because the effective components extracted under the conditions are more suitable to be used as the substrate components of the subsequent bidirectional fermentation.
Preferably, the cortex moutan and saussurea involucrata are pretreated before the extraction in the step (1), and the pretreatment comprises the following steps: drying, pulverizing and sieving.
Preferably, after the extraction in the step (1) is finished, the extracting solution is filtered and concentrated.
In the present invention, the basic medium in step (2) comprises: 1-5% of peptone, 1-5% of glucose, 0.5-2% of sodium chloride, 0.1-1% of calcium carbonate and the balance of water.
The mass concentration of the peptone can be 1%, 2%, 3%, 4%, 5% or the like; the mass concentration of the glucose can be 1%, 2%, 3%, 4%, 5% or the like; the mass concentration of the sodium chloride can be 0.5%, 1%, 1.5%, 2% or the like; the mass concentration of the calcium carbonate can be 0.1%, 0.2%, 0.4%, 0.5%, 0.6%, 0.8%, 1% or the like; other specific point values within the above range can be selected, and are not described in detail herein.
Preferably, the inoculation amount of the ganoderma lucidum mycelia in the step (2) in the basic culture medium is 4-6%, for example, 4%, 4.5%, 5%, 5.5%, or 6%, and other specific values in the above range can be selected, and are not described herein again.
The inoculation amount of the ganoderma lucidum mycelia in the basic culture medium obviously influences the efficacy of a final product, and when the inoculation amount is 4-6%, the ganoderma lucidum mycelia is more excellent in polysaccharide and flavone content, tyrosinase inhibition capability, hyaluronidase inhibition capability and DPPH and hydroxyl radical removal capability.
Preferably, the amount of the Chinese medicinal extract in step (2) is 4-6% of the mass of the basic culture medium inoculated with and cultured with ganoderma lucidum mycelia, for example, 4%, 4.5%, 5%, 5.5%, or 6%, and other specific values within the above range can be selected, and are not repeated herein.
The addition amount of the traditional Chinese medicine composition extract obviously influences the efficacy of a final product, and when the inoculation amount is 4-6%, the traditional Chinese medicine composition extract is more excellent in polysaccharide and flavone content, tyrosinase inhibition capability, hyaluronidase inhibition capability and DPPH and hydroxyl radical scavenging capability.
Preferably, the adding time of the Chinese medicinal extract in step (2) is 2-5 days of Ganoderma lucidum mycelium culture, such as 2 days, 3 days, 4 days or 5 days.
When the addition time of the traditional Chinese medicine combination extract is 2-5 days after the ganoderma lucidum mycelia are cultured, and more preferably 3 days, the efficacy of the finally obtained fermentation mycoplasm extract is better.
Preferably, the initial pH of the fermentation in step (2) is 5.5-6.5, such as pH =5.5, pH =6, or pH =6.5, and other specific values within the above range can be selected, and are not described in detail herein.
Preferably, the fermentation temperature in step (2) is 20-35 ℃, such as 20 ℃, 22 ℃,25 ℃, 28 ℃, 30 ℃, 32 ℃ or 35 ℃, and the like, the fermentation time is 5-14 days, such as 5 days, 7 days, 8 days, 10 days, 11 days, 12 days, 13 days or 14 days, and other specific values in the above range can be selected, and are not repeated herein.
The temperature and time and the initial pH environment of the preparation method of the present invention during the fermentation treatment also affect the efficacy of the finally obtained fermentation mycoplasm extract.
In the present invention, the solvent for the extraction in step (3) comprises water.
Preferably, the extraction temperature in step (3) is 20-30 ℃, for example 20 ℃, 22 ℃, 24 ℃,25 ℃, 26 ℃, 28 ℃ or 30 ℃ and the like, and the time is 1-3h, for example 1h, 1.5h, 2h, 2.5h or 3h and the like, and other specific values in the above range can be selected, and are not repeated herein.
Preferably, after the extraction in the step (3), the extracting solution is subjected to centrifugal filtration, concentration and drying to obtain the bidirectional fermentation mycoplasm extract of the traditional Chinese medicine composition.
As a preferred technical scheme of the present invention, the preparation method of the bidirectional fermentation mycoplasm extractive of the traditional Chinese medicine composition specifically comprises:
(1) Drying cortex moutan and herba Saussureae Involueratae at a mass ratio of (1-5) to 1, mixing, pulverizing, and sieving with 10-30 mesh sieve to obtain Chinese medicinal composition powder;
(2) Extracting the traditional Chinese medicine composition powder obtained in the step (1) by using 40-60% ethanol water solution with the mass concentration of 1 to 1 in a mass ratio of (8-12) to the raw materials at 70-90 ℃ for 1-3h for 1-3 times, filtering the extracting solution, and concentrating to the concentration of 0.1-1.0 g/mL to obtain a traditional Chinese medicine composition extract;
(3) Inoculating the activated ganoderma lucidum mycelia in a basic culture medium in an inoculation amount of 4-6%, and culturing for later use, wherein the basic culture medium comprises: 1-5% of peptone, 1-5% of glucose, 0.5-2% of sodium chloride, 0.1-1% of calcium carbonate and the balance of water;
(4) Adding the Chinese medicinal composition extract obtained in the step (2) into the culture medium which is cultured with the ganoderma lucidum mycelia for 2-5 days in the step (3), wherein the adding amount is 4-6%; fermenting at initial pH of 5.5-6.5 at 20-35 deg.C for 5-14 days, and filtering to obtain fermented mycoplasm;
(5) And (5) extracting the fermentation mycoplasm obtained in the step (4) with water at the temperature of 20-30 ℃ for 1-3h, centrifuging and filtering the extracting solution, concentrating, and drying to obtain the traditional Chinese medicine composition bidirectional fermentation mycoplasm extract.
On the other hand, the invention provides the traditional Chinese medicine composition bidirectional fermentation mycoplasm extract prepared by the preparation method of the traditional Chinese medicine composition bidirectional fermentation mycoplasm extract.
The traditional Chinese medicine composition prepared by the preparation method has high contents of polysaccharide and flavone in the bidirectional fermentation mycoplasm extract, strong inhibition capability on tyrosinase and hyaluronidase, and strong scavenging capability on DPPH and hydroxyl free radicals.
In another aspect, the invention provides an application of the bidirectional fermentation mycoplasm extractive of the traditional Chinese medicine composition in preparing cosmetics.
Preferably, the cosmetic comprises a mask, cream, lotion, essence or aqua.
Compared with the prior art, the invention has the following beneficial effects:
the invention specifically selects two traditional Chinese medicine components of tree peony bark and snow lotus, the two traditional Chinese medicine components are cooperated and matched, a traditional Chinese medicine combination extract obtained by extracting the traditional Chinese medicine combination extract is used as a fermentation substrate component, lucid ganoderma is used as a fermentation strain, two-way fermentation is carried out to obtain a mycoplasm component, and the mycoplasm component is further extracted to obtain a two-way fermentation mycoplasm extract of the traditional Chinese medicine composition. The polysaccharide content of the obtained traditional Chinese medicine combination bidirectional fermentation mycoplasm extract is high and can reach 306.2mg/g at most; the flavone content is high and can reach 49.88mg/g at most; the inhibition capacity to tyrosinase is strong, and the half inhibition amount can reach 0.517mg/mL at the minimum; the inhibition capacity to hyaluronidase is strong, and the half inhibition amount can reach 10.65mg/mL at the lowest; the scavenging ability to DPPH is strong, and the half-scavenging amount can reach 0.052mg/mL at the lowest; the scavenging ability to hydroxyl free radical is strong, and the half-scavenging amount can reach 1.006mg/mL at the lowest. It can be used as potential cosmetic whitening, anti-aging and anti-allergy raw materials, and is applied to cosmetic products.
Detailed Description
In order to further illustrate the technical means and effects of the present invention, the technical solutions of the present invention are further described below with reference to the preferred embodiments of the present invention, but the present invention is not limited to the scope of the embodiments.
The apparatus used in the following examples and comparative examples is as follows:
1. ultraviolet-visible spectrophotometer: model number UV-1750, available from Shimadzu corporation, japan;
2. precision electronic balance: type AE24, available from Starorius, switzerland;
3. a water bath kettle: the model is DSY-2-8, purchased from Beijing Guohua medical instrument factory;
4. a rotary evaporator: model RE-5205, purchased from Shanghai Rongya Biochemical apparatus factory;
5. constant temperature shaking table: THZ.98AB, available from Shanghai-constant technology instruments, inc.;
6. a low-speed centrifuge: model lc.4016, available from seiko scientific instruments ltd, zhongkaiki, anhui;
7. electric heating air blast drying oven: type SYIOIS.2, available from Sanshui scientific instruments, inc., tianjin;
8. vertical pressure steam sterilizer: TYAIB type 24/10, available from Ningbo Jiuxing medical devices, inc.;
9. an ultrasonic cleaner: KB2800, available from ultrasound instruments Inc., kunshan;
10. a pulverizer: model 800A, available from kunzhou likang electrical appliances ltd;
11. MO MODULYOD-230 Freeze dryer available from Thermo Fisher Sciebtific, USA.
Example 1
The embodiment provides a bidirectional fermentation mycoplasm extractive of a traditional Chinese medicine composition, and a preparation method of the bidirectional fermentation mycoplasm extractive comprises the following steps:
(1) Drying 40 parts of cortex moutan and 10 parts of herba Saussureae Involueratae, mixing, pulverizing, and sieving with 20 mesh sieve to obtain Chinese medicinal composition powder;
(2) Taking the Chinese medicinal composition powder obtained in the step (1) as a raw material, extracting with 50% ethanol water solution at the mass ratio of 10 to the raw material of 1 at 80 ℃ for 2h, filtering the extractive solution, and concentrating to obtain a Chinese medicinal composition extract, wherein the extraction time is 2 hours, and the concentration of the extractive solution is 0.5 g/mL;
(3) Inoculating the activated ganoderma lucidum mycelia in a basic culture medium in an inoculation amount of 5%, and culturing for later use, wherein the basic culture medium comprises: 2% of peptone, 2% of glucose, 1% of sodium chloride, 0.5% of calcium carbonate and the balance of water;
(4) Adding the Chinese medicinal composition extract obtained in the step (2) into the culture medium which is cultured with the ganoderma lucidum mycelia for 3 days in the step (3), wherein the adding amount is 5%; fermenting at initial pH of 6.5 at 28 deg.C for 10 days, and filtering to obtain fermented mycoplasm;
(5) And (4) carrying out countercurrent extraction on the fermentation mycoplasm obtained in the step (4) for 2 hours at the temperature of 25 ℃, and carrying out centrifugal filtration, concentration and freeze drying on an extracting solution to obtain the traditional Chinese medicine composition bidirectional fermentation mycoplasm extract.
Example 2
The embodiment provides a bidirectional fermentation mycoplasm extractive of a traditional Chinese medicine composition, and a preparation method of the bidirectional fermentation mycoplasm extractive comprises the following steps:
(1) Drying 20 parts of cortex moutan and 10 parts of herba Saussureae Involueratae, mixing, pulverizing, and sieving with 20 mesh sieve to obtain Chinese medicinal composition powder;
(2) Extracting the traditional Chinese medicine composition powder obtained in the step (1) by using an ethanol water solution with the mass concentration of 60% and the mass ratio of the traditional Chinese medicine composition powder to the raw materials of 8;
(3) Inoculating the activated ganoderma lucidum mycelia in a basic culture medium in an inoculation amount of 6%, and culturing for later use, wherein the basic culture medium comprises: 2% of peptone, 2% of glucose, 1% of sodium chloride, 0.5% of calcium carbonate and the balance of water;
(4) Adding the Chinese medicinal composition extract obtained in the step (2) into the culture medium which is cultured with the ganoderma lucidum mycelia for 3 days in the step (3), wherein the adding amount is 6%; fermenting at initial pH of 5.5 at 25 deg.C for 14 days, and filtering to obtain fermented mycoplasm;
(5) And (3) extracting the fermented mycoplasm obtained in the step (4) with water at the temperature of 20 ℃ for 3 hours in a water bath, centrifuging and filtering the extracting solution, concentrating, and drying to obtain the traditional Chinese medicine composition bidirectional fermented mycoplasm extract.
Example 3
The embodiment provides a bidirectional fermentation mycoplasm extractive of a traditional Chinese medicine composition, and a preparation method of the bidirectional fermentation mycoplasm extractive comprises the following steps:
(1) Drying 50 parts of moutan bark and 10 parts of saussurea involucrate, mixing, crushing, and sieving with a 20-mesh sieve to obtain traditional Chinese medicine composition powder;
(2) Taking the Chinese medicinal composition powder obtained in the step (1) as a raw material, extracting with an ethanol aqueous solution with a mass concentration of 40% and a mass ratio of 12 to the raw material of 1h at 90 ℃, for 3 times, filtering the extracting solution, and concentrating to a concentration of 0.3g/mL to obtain a Chinese medicinal composition extract;
(3) Inoculating the activated ganoderma lucidum mycelia in a basic culture medium in an inoculation amount of 4%, and culturing for later use, wherein the basic culture medium comprises: 2% of peptone, 2% of glucose, 1% of sodium chloride, 0.5% of calcium carbonate and the balance of water;
(4) Adding the traditional Chinese medicine composition extract obtained in the step (2) into the culture medium which is cultured with the ganoderma lucidum mycelia for 3 days in the step (3), wherein the adding amount is 4%; fermenting at initial pH of 6.5 at 35 deg.C for 6 days, and filtering to obtain fermented mycoplasm;
(5) And (4) carrying out countercurrent extraction on the fermentation mycoplasm obtained in the step (4) for 1h at the temperature of 30 ℃, and carrying out centrifugal filtration, concentration and drying on an extracting solution to obtain the traditional Chinese medicine composition bidirectional fermentation mycoplasm extract.
Example 4
This example provides a bidirectional fermented mycoplasm extractive of traditional Chinese medicine composition, and the preparation method is different from that of example 1 only in that in step (2), an ethanol aqueous solution with an extraction reagent mass concentration of 50% is replaced by water, and other conditions are kept unchanged.
Example 5
This example provides a bidirectional fermentation mycoplasm extract of a traditional Chinese medicine composition, which is prepared by a method different from that of example 1 only in that an ethanol aqueous solution with a mass concentration of 20% is replaced by an ethanol aqueous solution with a mass concentration of 50% in an extraction reagent in step (2), and other conditions are kept unchanged.
Example 6
This example provides a bidirectional fermented mycoplasm extractive of traditional Chinese medicine composition, and the preparation method is different from that of example 1 only in that in step (2), an ethanol aqueous solution with the extraction reagent mass concentration of 50% is replaced by an ethanol aqueous solution with the mass concentration of 70%, and other conditions are kept unchanged.
Example 7
This example provides a bidirectional fermentation fungal extract of a Chinese medicinal composition, which is prepared according to the method described in example 1, except that the inoculation amount of Ganoderma lucidum mycelia in step (3) is 3%, and other conditions are kept unchanged.
Example 8
This example provides a bidirectional fermented mycoplasm extract of a traditional Chinese medicine composition, and the preparation method of the bidirectional fermented mycoplasm extract of the traditional Chinese medicine composition is different from that of the traditional Chinese medicine composition in the example 1 only in that the inoculation amount of ganoderma lucidum mycelia in the step (3) is 4%, and other conditions are kept unchanged.
Example 9
This example provides a bidirectional fermented mycoplasm extract of a traditional Chinese medicine composition, and the preparation method of the bidirectional fermented mycoplasm extract of the traditional Chinese medicine composition is different from that of the traditional Chinese medicine composition in the example 1 only in that the inoculation amount of ganoderma lucidum mycelia in the step (3) is 6%, and other conditions are kept unchanged.
Example 10
The present example provides a bidirectional fermented mycoplasm extract of traditional Chinese medicine composition, and the preparation method is different from that of example 1 only in that the addition amount of the traditional Chinese medicine composition extract in step (4) is 1%, and other conditions are kept unchanged.
Example 11
The present example provides a bidirectional fermentation mycoplasm extract of a traditional Chinese medicine composition, and the preparation method is different from that of example 1 only in that the addition amount of the traditional Chinese medicine composition extract in the step (4) is 10%, and other conditions are kept unchanged.
Example 12
The present example provides a bidirectional fermentation mycoplasm extract of a traditional Chinese medicine composition, which is prepared by a method different from that in example 1 only in that the traditional Chinese medicine composition extract obtained in step (2) is added into a basal medium before ganoderma lucidum mycelia are inoculated, and other conditions are kept unchanged.
Example 13
This example provides a bidirectional fermentation mycoplasm extract of a traditional Chinese medicine composition, which is prepared by a method different from that of example 1 only in that the traditional Chinese medicine composition extract obtained in step (2) is added to a culture medium obtained in step (3) after ganoderma lucidum mycelia are cultured for 2 days in step (4), and other conditions are kept unchanged.
Example 14
The present example provides a bidirectional fermented mycoplasm extract of a traditional Chinese medicine composition, which is different from the preparation method in example 1 only in that the traditional Chinese medicine composition extract obtained in step (2) is added into the culture medium after the ganoderma lucidum mycelia are cultured for 5 days in step (3) in step (4), and other conditions are kept unchanged.
Example 15
This example provides a bidirectional fermentation mycoplasm extractive of traditional Chinese medicine composition, and the preparation method is different from that of example 1 only in that fermentation is carried out at an initial pH value of 5 in step (4), and other conditions are kept unchanged.
Example 16
This example provides a bidirectional fermentation mycoplasm extract of a Chinese medicinal composition, which is prepared by the method different from that of example 1 only in that fermentation is carried out at an initial pH of 7 in step (4), and other conditions are kept unchanged.
Example 17
This example provides a bidirectional fermentation mycoplasm extract of a Chinese medicinal composition, and the preparation method is different from that of example 1 only in that the fermentation temperature in step (4) is 20 ℃, and other conditions are kept unchanged.
Example 18
This example provides a bidirectional fermentation mycoplasm extract of a Chinese medicinal composition, and the preparation method is different from that of example 1 only in that the fermentation temperature in step (4) is 35 ℃, and other conditions are kept unchanged.
Example 19
This example provides a bidirectional fermentation mycoplasm extract of a Chinese medicinal composition, which is prepared by the method different from that of example 1 only in that the fermentation time in step (4) is 5 days, and other conditions are kept unchanged.
Example 20
This example provides a bidirectional fermentation mycoplasm extract of a Chinese medicinal composition, and the preparation method is different from that of example 1 only in that the fermentation time in step (4) is 14 days, and other conditions are kept unchanged.
Comparative example 1
The comparative example provides a traditional Chinese medicine composition extract, and the preparation method comprises the following steps:
(1) Drying 40 parts of cortex moutan and 10 parts of herba Saussureae Involueratae, mixing, pulverizing, and sieving with 20 mesh sieve to obtain Chinese medicinal composition powder;
(2) Taking the traditional Chinese medicine composition powder obtained in the step (1) as a raw material, extracting with an ethanol aqueous solution with the mass ratio of 10 to the raw material being 1 and the mass concentration being 50%, wherein the extraction temperature is 80 ℃, the extraction time is 2h, the extraction times are 2 times, filtering and concentrating the extracting solution to the concentration of 0.5g/mL, and freeze-drying to obtain the traditional Chinese medicine composition extract.
Comparative example 2
The comparative example provides a common ganoderma lucidum fermentation mycoplasm extractive, and the preparation method comprises the following steps:
(1) Inoculating the activated ganoderma lucidum mycelia in a basic culture medium in an inoculation amount of 5%, and culturing for later use, wherein the basic culture medium comprises: 2% of peptone, 2% of glucose, 1% of sodium chloride, 0.5% of calcium carbonate and the balance of water;
(2) Fermenting at initial pH of 6.5 at 28 deg.C for 13 days, and filtering to obtain fermented mycoplasm;
(3) And (3) carrying out countercurrent extraction on the fermentation mycoplasm obtained in the step (2) for 2 hours at 25 ℃, and carrying out centrifugal filtration, concentration and freeze drying on the extracting solution to obtain the common ganoderma lucidum fermentation mycoplasm extract.
Comparative example 3
The comparative example provides a cortex moutan bi-directional fermentation mycoplasm extract, and the preparation method comprises the following steps:
(1) Crushing 50 parts of moutan bark, and sieving with a 20-mesh sieve to obtain moutan bark powder;
(2) Taking the moutan bark powder obtained in the step (1) as a raw material, extracting with an ethanol water solution with a mass concentration of 50% and a mass ratio of 10 to the raw material of 1 at 80 ℃, wherein the extraction time is 2 hours, the extraction frequency is 2 times, and filtering and concentrating the extracting solution to a concentration of 0.5g/mL to obtain a moutan bark extract;
(3) Inoculating the activated ganoderma lucidum mycelia in a basic culture medium in an inoculation amount of 5%, and culturing for later use, wherein the basic culture medium comprises: 2% of peptone, 2% of glucose, 1% of sodium chloride, 0.5% of calcium carbonate and the balance of water;
(4) Adding the moutan bark extract obtained in the step (2) into the culture medium which is cultured with the ganoderma lucidum mycelia for 3 days in the step (3), wherein the addition amount is 5%; fermenting at initial pH of 6.5 at 28 deg.C for 10 days, and filtering to obtain fermented mycoplasm;
(5) And (4) carrying out countercurrent extraction on the fermentation mycoplasm obtained in the step (4) for 2 hours at 25 ℃, and carrying out centrifugal filtration, concentration and freeze drying on an extracting solution to obtain the tree peony bark bidirectional fermentation mycoplasm extract.
Comparative example 4
The comparative example provides a saussurea involucrate bidirectional fermentation mycoplasm extractive, and the preparation method comprises the following steps:
(1) Crushing 50 parts of saussurea involucrate, and sieving the crushed saussurea involucrate by a 20-mesh sieve to obtain saussurea involucrate powder;
(2) Extracting the saussurea involucrate powder obtained in the step (1) by using an ethanol water solution with the mass concentration of 50% and the mass ratio of the powder to the raw material of 10;
(3) Inoculating the activated ganoderma lucidum mycelia in a basic culture medium in an inoculation amount of 5%, and culturing for later use, wherein the basic culture medium comprises: 2% of peptone, 2% of glucose, 1% of sodium chloride, 0.5% of calcium carbonate and the balance of water;
(4) Adding the saussurea involucrate extract obtained in the step (2) into the culture medium which is cultured with the ganoderma lucidum mycelia for 3 days in the step (3), wherein the adding amount is 5%; fermenting at initial pH of 6.5 at 28 deg.C for 10 days, and filtering to obtain fermented mycoplasm;
(5) And (4) carrying out countercurrent extraction on the fermentation mycoplasm obtained in the step (4) for 2 hours at the temperature of 25 ℃, and carrying out centrifugal filtration, concentration and freeze drying on an extracting solution to obtain the saussurea involucrate bidirectional fermentation mycoplasm extract.
Evaluation test:
the products obtained in examples 1 to 20 and comparative examples 1 to 4 were dissolved in 99 times by weight of purified water, and the products were subjected to the measurement of the contents of polysaccharides and total flavonoids, the measurement of DPPH free radical scavenging ability, the measurement of hydroxyl free radical scavenging ability, the measurement of tyrosinase inhibitory ability, and the measurement of hyaluronidase inhibitory ability, respectively, by ultraviolet spectrophotometry.
(1) The DPPH free radical scavenging ability is detected as follows:
(1.1) accurately weighing 47.7g of DPPH reagent, metering to a 100mL brown volumetric flask, taking 5mL of the brown volumetric flask, metering to a 50mL brown volumetric flask by using absolute ethyl alcohol, and obtaining DPPH test solution (47.7 mg/L). 2mL of the DPPH test solution was removed, and 0.1mL of the product obtained in examples 1 to 20 and comparative examples 1 to 4 was added as shown in the following table, and an appropriate amount of anhydrous ethanol was added to equalize the total mass. Standing in dark for 30min after mixing, measuring absorbance at 517nm with anhydrous ethanol as blank control, and calculating DPPH clearance of each extractive solution according to the following formula:
DPPH clearance (%) = [1- (As-Ar)/A0 ]. Times.100%
In the formula: as is the absorbance of the reaction system of the extract and DPPH; ar-absorbance of extract-absolute ethanol; a0-absorbance of blank control in absolute ethanol.
(1.2) measurement of half-scavenging amount of DPPH free radical in extract
And respectively measuring the clearance of the extracts with different concentrations, taking the concentration as an abscissa and the clearance as an ordinate to make a straight line, and calculating the concentration of the extract when the clearance is 50 percent to obtain the half clearance of DPPH free radicals after solving a straight line equation.
(2) The method for detecting the scavenging capacity of the hydroxyl free radical comprises the following steps:
(2.1) adding 1.0mL of FeSO into the reaction system 4 Solution (9.0 mmol/L), 1mL salicylic acid-ethanol solution (9.0 mmol/mL), 1mL samples, then 1mL H 2 O 2 (8.8 mmol/mL), reacting in 37 ℃ water bath for 30min, taking absolute ethyl alcohol as a blank control, measuring the absorbance at 510nm, and calculating the removal rate of OH of each extracting solution according to the following formula:
OH clearance (%) = [ (A0-Ax 0)/A0 ]. Times 100%
In the formula: ax is the absorbance of an extracting solution reaction system; ax 0-without addition of H 2 O 2 Absorbance of the sample (absolute ethanol instead of H) 2 O 2 ) (ii) a A0-absorbance of blank control in absolute ethanol.
(2.2) measurement of half-scavenging amount of hydroxyl radical in extract
And respectively measuring the clearance of the extracts with different concentrations, taking the concentration as an abscissa and the clearance as an ordinate to make a straight line, and calculating the concentration of the extract when the clearance is 50 percent to be the half clearance of the hydroxyl free radicals after solving a straight line equation.
(3) The tyrosinase inhibition ability was measured as follows:
(3.1) preparation of L-tyrosinase solution and tyrosinase solution
Weighing 0.90595g of L-tyrosine, using PBS buffer solution to fix the L-tyrosine in a 100mL volumetric flask, and then diluting by 10 times to obtain 5mM L-tyrosine solution (the L-tyrosine can be dissolved by a small amount of hydrochloric acid firstly and then fixed by the PBS buffer solution). Tyrosinase, diluted to 100U/mL with deionized water.
(3.2) determination of tyrosinase inhibition Rate of extract
X mL of an extract solution (1.4-X) mL of 0.05mol/L phosphate buffer (pH = 6.8) was transferred to a cuvette by a micropipette, 0.8mL of 0.5mmol/L tyrosine solution was added thereto, and finally 0.40mL of tyrosinase solution was added thereto, followed by rapid mixing, measurement of a time-dependent increase curve of an optical density value at a wavelength of 475nm at a constant temperature of 30 ℃ and determination of a slope inhibition ratio from the curve.
(3.3) extract tyrosinase IC50 assay
Respectively measuring the inhibition rates of the extracts with different concentrations, taking the concentration as an abscissa and the inhibition rate as an ordinate to form a straight line, and calculating the concentration of the extract when the inhibition rate is 50 percent as the IC50 after solving a straight line equation.
(4) The hyaluronidase inhibition was measured as follows:
(4.1) 0.1mL of CaCl was taken 2 Reacting the solution (0.25 mmol/L) and 0.5mL of hyaluronidase solution (100U/mL) in water bath at 37 ℃ for 20min; adding 0.5mL of the extracting solution, and keeping the temperature for 20min; then adding 0.5mL of sodium hyaluronate solution (0.5 g/L), reacting for 30min, taking out and cooling for 5min; adding 0.1mL of sodium hydroxide solution (0.4 mol/L) and 0.5mL of acetylacetone solution (3.5 mL of acetylacetone dissolved in 50mL of 1.0mol/L sodium carbonate solution), boiling in water bath for 15min, and immediately transferring to ice water bath for 5min; 1mL of Ellisib reagent (0.8 g p-dimethylaminobenzaldehyde dissolved in 15mL concentrated hydrochloric acid and 15mL absolute ethanol) was added dropwise, diluted with 3mL absolute ethanol, left at 25 ℃ for 20min for color development, and the absorbance was measured at 540nm with dipotassium glycyrrhizinate as a control. The hyaluronidase inhibition rate was calculated according to the following formula:
hyaluronidase inhibition = { [ (A-B) - (C-D) ]/(A-B) } × 100%
In the formula: a-absorbance of control solution (the sample solution was replaced with acetic acid buffer solution); b-the absorbance of the control blank solution (the sample solution and the enzyme solution are replaced by acetic acid buffer solution); c-absorbance of the sample solution; d-absorbance of sample blank solution (acetic acid buffer solution was used instead of enzyme solution).
(4.2) extract hyaluronidase IC50 assay
Respectively measuring the inhibition rate of the extracts with different concentrations, taking the concentration as an abscissa and the inhibition rate as an ordinate to make a straight line, and calculating the concentration of the extract when the inhibition rate is 50 percent to obtain the IC50 after solving a straight line equation.
(5) The detection method of the polysaccharide content comprises the following steps:
(5.1) preparing an anthrone reagent: accurately weighing 0.1g anthrone, adding 80% concentrated H 2 SO 4 Dissolve 100mL and shake well. Preparing a glucose standard solution: mixing the anhydrous grapePutting the sugar in a phosphorus pentoxide dryer, precisely weighing 100mg after 12h, adding the sugar into a beaker, adding a small amount of distilled water for dilution, transferring the diluted solution into a 100mL volumetric flask, washing the beaker with distilled water for 1-2 times, transferring the washing solution into the volumetric flask, adding distilled water to the scale, and shaking up.
(5.2) preparation of glucose Standard Curve: taking 7 supports to plug the test tube, and precisely preparing a series of glucose solutions with different concentrations according to the data in the following table:
adding 6.0mL of anthrone reagent into each test tube, shaking, mixing, and heating in boiling water bath for 15min. Taking out, and rapidly soaking in ice water bath for cooling for 15min. The absorbance of each of the remaining tubes was rapidly measured in a blank of tube No. 0 at a wavelength of 625 nm. And drawing a standard curve by taking the standard glucose content (g) as an abscissa and the light absorption value as an ordinate.
(5.3) adjusting the sugar concentration of the sample solution to a measurement range, accurately sucking 2.0mL of the sample solution, placing the sample solution in a dry clean test tube, immediately adding 6.0mL of anthrone reagent, uniformly mixing by oscillation, placing the test tubes in a boiling water bath, and heating for 15min. Taking out, rapidly soaking in ice water bath for cooling for 15min, and repeating each concentration for 3 times. The absorbance of each tube was rapidly measured at a wavelength of 625 nm. And calculating the concentration of total sugar in each sample solution according to the standard curve of the glucose content and the light absorption value of the sample solution, and calculating the total sugar content.
(6) The detection method of the total flavone content comprises the following steps:
(6.1) preparing a 5% sodium nitrite solution: 5.0g of sodium nitrite (NaNO) are weighed out 2 ) Adding the solid into a beaker, adding a small amount of distilled water for dilution, transferring the diluted solution into a 100mL volumetric flask, washing the beaker with distilled water for 2 times, transferring the washing solution into the volumetric flask, adding distilled water to the scale, and shaking up. Preparing a 10% aluminum nitrate solution: 10.0g of aluminum nitrate solid was weighed, added to a beaker, diluted with a small amount of distilled water, and the diluted solution was transferred to a 100mL containerAnd (4) washing the beaker 1-2 times in a measuring flask by using distilled water, transferring the washing liquid into a volumetric flask, adding the distilled water to the scale mark, and shaking up. Preparing a 5% sodium hydroxide solution: weighing 5.0g of sodium hydroxide solid, adding the sodium hydroxide solid into a beaker, adding a small amount of distilled water for dilution, transferring the diluted solution into a 100mL volumetric flask, washing the beaker with distilled water for 2 times, transferring the washing solution into the volumetric flask, adding distilled water to the scale, and shaking up. Preparing a rutin standard solution: accurately weighing 10.0mg of rutin standard substance, adding the same solvent as the sample to be measured into a 50mL measuring flask, and adding to the scale to obtain a rutin standard substance solution (0.20 mg/mL).
(6.2) drawing of standard curve: taking rutin standard substance solution 0, 0.05, 0.10, 0.20, 0.50, 1.0, 1.2 and 1.5mL to 5mL volumetric flasks, adding a solvent to 2.0mL and 5% sodium nitrite solution 0.2mL, shaking uniformly, standing for 6min, adding 10% aluminum nitrate solution 0.2mL, shaking uniformly, standing for 6min, adding 5% sodium hydroxide solution 1.0mL, adding a solvent to the scale, and shaking uniformly to obtain standard substance test solution. The corresponding solvent was used as a blank. And (3) measuring the absorbance of the solution at the wavelength of 510nm, drawing a standard curve by taking the absorbance as a vertical coordinate and the concentration of rutin as a horizontal coordinate, and solving a standard linear equation.
(6.3) measurement of sample solution: adjusting the total flavone concentration of the sample solution to a measuring range, accurately sucking 2.0mL of the sample solution, placing the sample solution in a dry clean test tube, adding 0.2mL of 5% sodium nitrite solution, shaking uniformly, placing for 5min, adding 0.2mL of 10% aluminum nitrate solution, shaking uniformly, placing for 6min, adding 1.0mL of 5% sodium hydroxide solution, adding a solvent to the scale, and shaking uniformly to obtain a sample test solution. 3 replicates were made for each concentration. The sample solution was used to rapidly determine the absorbance of each tube at a wavelength of 510 nm. Calculating the concentration of total flavonoids according to the standard curve of rutin solution and the light absorption value of the sample solution, wherein the total flavonoids content in the sample is rutin (C) 27 H 30 O 16 ) And (4) showing.
The results are shown in table 1:
TABLE 1
As can be seen from Table 1: the polysaccharide content of the bidirectional fermentation mycoplasm extractive of the traditional Chinese medicine composition is high and can reach 306.2mg/g at most; the flavone content is high and can reach 49.88mg/g at most; the inhibition capacity to tyrosinase is strong, and the half inhibition amount can reach 0.517mg/mL at the lowest; the inhibition capacity to hyaluronidase is strong, and the half inhibition amount can reach 10.65mg/mL at the lowest; the scavenging ability to DPPH is strong, and the half-scavenging amount can reach 0.052mg/mL at the lowest; has strong scavenging ability to hydroxyl free radical, and the half-scavenging amount can reach 1.006mg/mL at the lowest. The effects are all better than those of the ganoderma lucidum fermentation mycoplasm extractive without the traditional Chinese medicine combined extractive or the bidirectional fermentation mycoplasm extractive with only one of peony and snow lotus.
Application example 1
The present application provides a skin care composition having the formula as shown in the following table:
| component name | Mass percentage content (%) |
| Deionized water | Allowance of |
| Bidirectional fermentation mycoplasm extract prepared in example 1 | 10 |
| Methyl glucitol polyether-10 | 3 |
| Glycerol | 2 |
| Arbutin | 1.5 |
| Polysorbate-60 | 1.5 |
| Urea | 0.5 |
| PCA sodium salt | 0.5 |
| Essence | 0.2 |
| Iodoproparganol butyl carbamate | 0.2 |
| Hydroxy phenyl methyl ester | 0.1 |
| Propyl hydroxybenzoate | 0.05 |
| Menthol | 0.05 |
| EDTA disodium salt | 0.05 |
Application comparative example 1
The application comparative example provides a skin care composition, and the formula composition of the skin care composition is shown in the following table:
evaluation test:
50 volunteers, 20-45 years old, half of men and half of women, were selected. The skin care composition prepared in application example 1 was applied to the left side, the skin care composition prepared in application example 1 was applied to the right side, the skin care composition was applied once a day in the morning and evening, 2g of the product was applied to the face each time, the face was massaged for 2 minutes, and the skin whitening and tightening effects of the left and right faces were scored on the volunteers for 30 days, respectively, with a maximum score of 5 and a minimum score of 1, and the evaluation results are shown in table 2 (average score):
TABLE 2
| Evaluation item | Application example 1 | Application comparative example 1 |
| Whitening ability | 4.3 | 3.3 |
| Capability of tightening | 4.1 | 3.5 |
As can be seen from table 2: the traditional Chinese medicine composition bidirectional fermentation mycoplasm extract disclosed by the invention is added into a cosmetic product, so that the sensory scores of skin whiteness and delicacy are obviously improved.
The applicant states that the present invention is described by the above embodiments, but the present invention is not limited to the above embodiments, i.e. the present invention is not limited to the above embodiments. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.
The preferred embodiments of the present invention have been described in detail, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are all within the protection scope of the present invention.
It should be noted that, in the above embodiments, the various features described in the above embodiments may be combined in any suitable manner, and in order to avoid unnecessary repetition, the present invention does not separately describe various possible combinations.
Claims (11)
1. A preparation method of a traditional Chinese medicine composition bidirectional fermentation mycoplasm extract with whitening, anti-aging and anti-allergy effects is characterized by comprising the following steps:
(1) Extracting cortex moutan and herba Saussureae Involueratae at a mass ratio of (1-5): 1 with 40-60% ethanol water solution as solvent to obtain Chinese medicinal extract; the extraction temperature is 70-90 ℃, the extraction time is 1-3h, and the extraction times are 1-3;
(2) Adding the Chinese medicinal extract obtained in step (1) into a basic culture medium inoculated with and cultured with Ganoderma mycelia for 2-5 days of Ganoderma mycelia culture, fermenting at 20-35 deg.C with initial pH of 5.5-6.5 for 6-14 days, and filtering to obtain fermented mycoplasm; the inoculation amount of the ganoderma lucidum mycelia in the basic culture medium is 4-6%, and the addition amount of the traditional Chinese medicine composition extract is 4-6% of the mass of the basic culture medium inoculated with and cultured with the ganoderma lucidum mycelia;
(3) And (3) extracting the fermentation mycoplasm obtained in the step (2) to obtain the traditional Chinese medicine composition bidirectional fermentation mycoplasm extract.
2. The preparation method of the bidirectional fermentation mycoplasm extractive with the effects of whitening, anti-aging and anti-allergy as claimed in claim 1, wherein the mass ratio of the ethanol water solution to the raw materials is (8-12) to 1.
3. The method for preparing the bidirectional fermentation mycoplasm extractive of the traditional Chinese medicine composition with the effects of whitening, anti-aging and anti-allergy as claimed in claim 1, wherein the cortex moutan radicis and the saussurea involucrata are pretreated before the extraction in the step (1), and the pretreatment comprises the following steps: drying, pulverizing and sieving.
4. The method for preparing the bidirectional fermentation mycoplasm extractive of the traditional Chinese medicine composition with the effects of whitening, resisting aging and resisting allergy as claimed in claim 1, wherein the extract is filtered and concentrated after the extraction in the step (1) is completed.
5. The method for preparing the bidirectional fermentation mycoplasm extractive of the traditional Chinese medicine composition with the effects of whitening, anti-aging and anti-allergy as claimed in claim 1, wherein the basic culture medium in the step (2) comprises: 1-5% of peptone, 1-5% of glucose, 0.5-2% of sodium chloride, 0.1-1% of calcium carbonate and the balance of water.
6. The method for preparing the bidirectional fermentation mycoplasm extractive of the traditional Chinese medicine composition with the effects of whitening, anti-aging and anti-allergy as claimed in claim 1, wherein the solvent extracted in the step (3) comprises water.
7. The method for preparing the bidirectional fermented mycoplasm extractive with the effects of whitening, resisting aging and resisting allergy as claimed in claim 1, wherein the extraction temperature in the step (3) is 20-30 ℃ and the extraction time is 1-3 h.
8. The method for preparing the traditional Chinese medicine composition bidirectional fermentation mycoplasm extract with the effects of whitening, anti-aging and anti-allergy as claimed in claim 1, wherein the traditional Chinese medicine composition bidirectional fermentation mycoplasm extract is obtained by performing centrifugal filtration, concentration and drying on an extracting solution after the extraction in the step (3).
9. The traditional Chinese medicine composition bidirectional fermentation mycoplasm extract with whitening, anti-aging and anti-allergy effects prepared by the preparation method of the traditional Chinese medicine composition bidirectional fermentation mycoplasm extract with whitening, anti-aging and anti-allergy effects of any one of claims 1 to 8.
10. The use of the bi-directional fermentation fungal extract of claim 9 for preparing cosmetics with whitening, anti-aging, and anti-allergy effects.
11. The use of claim 10, wherein the cosmetic comprises a mask, cream, lotion, serum, or aqua.
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