A plant fermentation composition for preventing hair loss and promoting hair growth, and its preparation method and application

Abstract

The present invention discloses a plant fermentation composition for preventing hair loss and promoting hair growth, and a preparation method and application thereof. The plant fermentation composition uses only angelica, Polygonum multiflorum and Platycladus orientalis leaves as plant components. Different from the common direct use of extracts as medicine, the composition of the present invention is firstly water-extracted with angelica, and then treated with enzymes to obtain an enzymatic extract of angelica, and water-extracted with Polygonum multiflorum and Platycladus orientalis, and then treated with enzymes to obtain an enzymatic extract of Polygonum multiflorum and Platycladus orientalis leaves; after the two enzymatic extracts are combined, they are co-fermented with lactic acid bacteria to obtain a plant fermentation composition. After experimental verification, after the three plants are subjected to enzymatic extraction and fermentation composite treatment, the content of the active ingredients in the three plants, namely angelica polysaccharides, diphenylethylene glycosides, and Platycladus orientalis leaf flavonoids, is significantly increased, and there is less precipitation, the stability is significantly improved, and the stickiness is significantly reduced. Compared with the anti-hair loss and hair growth products mixed with general plant extracts, the plant fermentation composition has a better anti-hair loss effect, a higher hair density, and only three plant components, so it is safer.

Description

Technical Field

[0001] The invention relates to the technical field of preventing hair loss and promoting hair growth, and in particular to a plant fermentation composition for preventing hair loss and promoting hair growth, and a preparation method and application thereof. Background Art

[0002] With the acceleration of the pace of life and the continuous increase of work intensity, androgenic alopecia (seborrheic alopecia), alopecia areata and other alopecia diseases are increasing, and they are becoming younger. This type of alopecia hinders the appearance and affects the appearance, and is more likely to bring huge mental stress and psychological burden to patients, which seriously affects the quality of life. In the treatment of alopecia, the commonly used Western medicines in the clinic are minoxidil and finasteride, but both drugs are prone to recurrence after withdrawal, and can also cause side effects such as skin irritation, scalp itching, hirsutism on the face and hands, dizziness, anorexia, tachycardia. In addition, although hair transplantation has a certain therapeutic effect, it is also limited in its clinical application because of the large surgical trauma, high cost and the need for multiple operations. The effect of non-drug treatments such as acupuncture is also not ideal.

[0003] In recent years, there have been more and more reports on the use of plant extracts to treat hair loss. However, a single Chinese herbal medicine has limited effect on promoting hair growth in vitro, and according to the theory of complementarity in traditional Chinese medicine, the use of Chinese herbal medicine compounding is effective. Existing plant-based anti-hair loss and hair growth products often contain more than a dozen plant ingredients, which is too expensive, and the safety of multiple ingredients is unknown. Summary of the invention

[0004] In order to overcome the shortcomings of the prior art, one of the objects of the present invention is to provide a method for preparing a plant fermentation composition for preventing hair loss and promoting hair growth. The plant raw materials used in the composition are only Angelica sinensis, Polygonum multiflorum and Platycladus orientalis leaves. After the plant raw materials are subjected to water extraction and enzymatic hydrolysis, the enzymatic hydrolyzate is mixed and co-fermented with probiotics to obtain the plant fermentation composition. The steps are simple and suitable for large-scale production; the second object of the present invention is to provide a plant fermentation composition for preventing hair loss and promoting hair growth, which has significantly improved active ingredient content, improved stability, reduced stickiness, better feel, and significantly improved anti-hair loss and hair growth effect; the third object of the present invention is to provide an application of a plant fermentation composition for preventing hair loss and promoting hair growth, and the plant fermentation composition is used to prepare anti-hair loss and hair growth products.

[0005] One of the purposes of the present invention is achieved by the following technical solution:

[0006] A method for preparing a plant fermentation composition for preventing hair loss and promoting hair growth comprises the following steps:

[0007] 1) extracting angelica sinensis with water, adding enzyme for treatment, killing the enzyme, filtering the supernatant to obtain angelica sinensis enzymolysis solution; extracting Polygonum multiflorum and Platycladus orientalis leaves with water, adding enzyme for treatment, killing the enzyme, filtering the supernatant to obtain Polygonum multiflorum and Platycladus orientalis leaf composite enzymolysis solution;

[0008] 2) combining the enzymatic hydrolyzate of Angelica sinensis and the composite enzymatic hydrolyzate of Polygonum multiflorum and Platycladus orientalis leaves, inoculating microorganisms for co-fermentation, filtering and collecting the supernatant to obtain a plant fermentation composition.

[0009] Furthermore, the mass ratio of one of angelica, Polygonum multiflorum and Platycladus orientalis to water is 1:(5-20), preferably the mass ratio of plant to water is 1:10. The mass ratio of angelica, Polygonum multiflorum and Platycladus orientalis is (0.5-2):(2-4):(4-6).

[0010] Furthermore, the water extraction temperature is 85-90°C and the time is 30-60min.

[0011] Further, in step 1), after the angelica is extracted with water, amylase is added and the mixture is treated at 40-60°C for 1-3h, the enzyme is inactivated at 80-90°C for 20min-60min, and the supernatant is filtered and collected to obtain an angelica hydrolyzate. Since angelica contains a lot of starch, amylase is used to remove the starch; after the Polygonum multiflorum and Platycladus orientalis leaves are extracted with water, pectinase is added and the mixture is treated at 40-60°C for 1-3h, the enzyme is inactivated at 80-90°C for 20min-60min, and the supernatant is filtered and collected to obtain a composite enzymatic hydrolyzate of Polygonum multiflorum and Platycladus orientalis leaves. Polygonum multiflorum and Platycladus orientalis leaves contain a lot of colloid, so pectinase is used to remove the colloid.

[0012] Furthermore, in step 2), the fermentation conditions are: fermentation at 35-38° C. for 18-36 hours; wherein the inoculation amount of the co-fermentation microorganism is 2-4% of the mass of the enzymatic hydrolysate.

[0013] Furthermore, the microorganism is lactic acid bacteria and / or yeast, preferably lactic acid bacteria.

[0014] Furthermore, in step 2), the supernatant obtained by centrifugation after fermentation is concentrated to a crude drug concentration of (0.1-0.2) g / g, thereby obtaining a plant fermentation composition for preventing hair loss and promoting hair growth. Preferably, a preservative needs to be added to the plant fermentation composition.

[0015] The second object of the present invention is achieved by adopting the following technical solution:

[0016] A plant fermentation composition for preventing hair loss and promoting hair growth is prepared by the method for preparing the plant fermentation composition for preventing hair loss and promoting hair growth.

[0017] The third object of the present invention is achieved by adopting the following technical solution:

[0018] An application of a plant fermentation composition for preventing hair loss and promoting hair growth, wherein the plant fermentation composition for preventing hair loss and promoting hair growth is used to prepare a hair loss and promoting hair growth product; wherein the mass percentage of the plant fermentation composition for preventing hair loss and promoting hair growth in the hair loss and promoting hair growth product is 3-5%.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] (1) The plant components used in the preparation method of the plant fermentation composition of the present invention are only angelica, Polygonum multiflorum and Platycladus orientalis leaves. Different from the common direct use of extracts as medicine, the preparation method of the present invention is to first extract angelica alone with water, add amylase to obtain angelica hydrolyzate; extract Polygonum multiflorum and Platycladus orientalis leaves with water, add pectinase to obtain Polygonum multiflorum Platycladus orientalis composite hydrolyzate, mix the angelica hydrolyzate and Polygonum multiflorum Platycladus orientalis leaf hydrolyzate, and then inoculate microorganisms for fermentation to obtain the plant fermentation composition. After experimental verification, the three plant extracts are subjected to enzymatic hydrolysis and fermentation composite treatment, the content of the active ingredients in the three plants, namely angelica polysaccharides, diphenylethylene glycosides, and Platycladus orientalis leaf flavonoids, is significantly increased, and the stability is compared with the fermentation composition that has only been fermented, with less precipitation, significantly improved stability, significantly reduced stickiness, and better use feel. Compared with the anti-hair loss and hair growth products mixed with general plant extracts, the plant fermentation composition has a better anti-hair loss effect and a higher hair growth density, and has only three plant components, so it is safer.

[0021] (2) The plant fermentation composition of the present invention can be added as one of the components of an anti-hair loss essence at a ratio of 3 to 5% by mass. After the essence is applied externally, experimental tests show that the number of hair loss in the subjects is reduced and the hair density and homogeneity are significantly improved, thus proving that it has a good anti-hair loss and hair growth effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is a graph showing the mean value of hair loss counts for the anti-hair loss and hair growth essence of Example 1;

[0023] Figure 2 This is a graph showing the mean hair density of the anti-hair loss and hair growth essence of Example 1. DETAILED DESCRIPTION

[0024] The present invention is further described below in conjunction with the accompanying drawings and specific implementation methods. It should be noted that, under the premise of no conflict, the various embodiments or technical features described below can be arbitrarily combined to form a new embodiment.

[0025] The plant raw materials selected for the plant fermentation composition for preventing hair loss and promoting hair growth of the present invention are angelica sinensis, Polygonum multiflorum and Platycladus orientalis leaves. From the pharmacological analysis of traditional Chinese medicine, Polygonum multiflorum and angelica sinensis as a plant pair can play the role of nourishing the liver and kidney and nourishing blood and promoting blood circulation. On the one hand, it can nourish blood, and the internal organs and skin and hair can be nourished when the blood vessels are full, and the hair is shiny and not easy to fall off; on the other hand, it can promote blood circulation, make blood flow unobstructed, not form stasis, nourish hair, strengthen hair roots, and grow new hair. And diphenylethylene glycoside is a quantitative detection index of Polygonum multiflorum, which has the effect of anti-oxidation and scavenging free radicals. Angelica sinensis polysaccharide (ASP), as one of the main components of angelica sinensis to exert pharmacological activity, has hematopoietic activity, antioxidant activity, immunomodulatory activity, antibacterial and anti-inflammatory effects. The effect of Platycladus orientalis leaves is to cool blood and stop bleeding, relieve cough and expectorant, eliminate rheumatism, and disperse swelling and poison. It is recorded in "Rihuazi": "burn the juice, apply it on the head, and the hair will be black and moisturized." It can be seen that in addition to the commonly used effect of cooling blood and stopping bleeding, Platycladus orientalis leaves can also be used to treat hair diseases. Platycladus orientalis leaf flavonoids, as the main active ingredient of Platycladus orientalis leaves, have the effects of relieving cough and reducing phlegm, relieving asthma, calming and relieving spasms, resisting pathogenic microorganisms, and anti-oxidation. All three have antioxidant and anti-inflammatory effects and can effectively improve the scalp microenvironment. It is considered that this may achieve the effect of blackening hair by jointly reducing melanocyte damage.

[0026] Example 1

[0027] A method for preparing a plant fermentation composition for preventing hair loss and promoting hair growth comprises the following steps:

[0028] 1) Wash and chop three plants of angelica sinensis, Polygonum multiflorum and Platycladus orientalis leaves in a mass ratio of 1:3:5, and then extract them respectively by water extraction, wherein the mass ratio of plant to water is 1:10; the water extraction temperature is 85°C, and the time is 30 minutes, to obtain angelica sinensis extract, Polygonum multiflorum extract and Platycladus orientalis leaf extract;

[0029] 2) adding amylase to the angelica extract and treating it at 50°C for 2h, inactivating the enzyme at 85°C for 30min, filtering and collecting the supernatant to obtain an angelica hydrolyzate; the amylase accounts for 0.1% of the weight of the angelica extract; adding pectinase to the mixture of the Polygonum multiflorum extract and the Platycladus orientalis leaf extract, treating it at 50°C for 2h, inactivating the enzyme at 85°C for 30min, filtering and collecting the supernatant to obtain a Polygonum multiflorum and Platycladus orientalis leaf composite enzymatic hydrolyzate; the pectinase accounts for 0.2% of the total weight of the Polygonum multiflorum extract and the Platycladus orientalis leaf extract;

[0030] 3) After combining the angelica enzymatic hydrolysate and the Polygonum multiflorum and Platycladus orientalis leaf composite enzymatic hydrolysate, inoculating microorganisms, fermenting at 37° C. and 200 rpm using a baffled shake flask, sealing with 8 layers of gauze to increase dissolved oxygen, fermenting for 24 hours, filtering the supernatant, concentrating it to a crude drug concentration of 0.15 g / g, adding a preservative, and obtaining a plant fermentation composition for preventing hair loss and promoting hair growth; wherein the inoculation amount of the microorganism is 4% of the mass of the enzymatic hydrolysate, and the inoculated microorganism is lactic acid bacteria.

[0031] Example 2

[0032] The method for preparing the plant fermentation composition for preventing hair loss and promoting hair growth in this embodiment is different from that in Embodiment 1 in that the co-fermentation microorganism selected in step 3) is yeast.

[0033] Example 3

[0034] A method for preparing a plant fermentation composition for preventing hair loss and promoting hair growth comprises the following steps:

[0035] 1) Wash and chop three plants of angelica sinensis, Polygonum multiflorum and Platycladus orientalis leaves in a mass ratio of 0.5:2:4, and then extract them respectively by water extraction, wherein the mass ratio of plant to water is 1:5; the water extraction temperature is 85°C, and the time is 60 minutes, to obtain angelica sinensis extract, Polygonum multiflorum extract and Platycladus orientalis leaf extract;

[0036] 2) adding amylase to the angelica extract and treating it at 40°C for 3h, inactivating the enzyme at 80°C for 20min, filtering and collecting the supernatant to obtain an angelica hydrolyzate; the amylase accounts for 0.1% of the weight of the angelica extract; after mixing the Polygonum multiflorum extract and the Platycladus orientalis leaf extract, adding pectinase to treat it at 40°C for 1h, inactivating the enzyme at 80°C for 60min, filtering and collecting the supernatant to obtain Polygonum multiflorum and Platycladus orientalis leaf composite enzymatic hydrolyzates; the pectinase accounts for 0.2% of the total weight of the Polygonum multiflorum extract and the Platycladus orientalis leaf extract respectively;

[0037] 3) After combining the angelica enzymatic hydrolysate and the Polygonum multiflorum and Platycladus orientalis leaf composite enzymatic hydrolysate, inoculating microorganisms, fermenting at 35° C. and 200 rpm using a baffled shake flask, sealing with 8 layers of gauze to increase dissolved oxygen, fermenting for 18 hours, filtering the supernatant, concentrating it to a crude drug concentration of 0.1 g / g, adding a preservative, and obtaining a plant fermentation composition for preventing hair loss and promoting hair growth; wherein the inoculation amount of the microorganism is 2% of the mass of the enzymatic hydrolysate, and the inoculated microorganism is a yeast.

[0038] Example 4

[0039] A method for preparing a plant fermentation composition for preventing hair loss and promoting hair growth comprises the following steps:

[0040] 1) Wash and chop three plants of angelica sinensis, Polygonum multiflorum and Platycladus orientalis leaves in a mass ratio of 2:4:6, and then extract them respectively by water extraction, wherein the mass ratio of plant to water is 1:20; the water extraction temperature is 90°C, and the time is 60 minutes, to obtain angelica sinensis extract, Polygonum multiflorum extract and Platycladus orientalis leaf extract;

[0041] 2) adding amylase to the angelica extract and treating it at 60°C for 3h, inactivating the enzyme at 90°C for 20min, filtering and collecting the supernatant to obtain an angelica hydrolyzate; the amylase accounts for 0.1% of the weight of the angelica extract; after mixing the Polygonum multiflorum extract and the Platycladus orientalis leaf extract, adding pectinase to treat it at 60°C for 1h, inactivating the enzyme at 90°C for 20min, filtering and collecting the supernatant to obtain Polygonum multiflorum and Platycladus orientalis leaf composite enzymatic hydrolyzates; the pectinase accounts for 0.2% of the total weight of the Polygonum multiflorum extract and the Platycladus orientalis leaf extract respectively;

[0042] 3) After combining the angelica enzymatic hydrolysate and the Polygonum multiflorum and Platycladus orientalis leaf composite enzymatic hydrolysate, inoculating microorganisms, fermenting at 38° C. and 200 rpm using a baffled shake flask, sealing with 8 layers of gauze to increase dissolved oxygen, fermenting for 18 hours, filtering the supernatant, concentrating it to a crude drug concentration of 0.2 g / g, adding a preservative, and obtaining a plant fermentation composition for preventing hair loss and promoting hair growth; wherein the inoculation amount of the microorganism is 3% of the mass of the enzymatic hydrolysate, and the inoculated microorganism is lactic acid bacteria.

[0043] Comparative Example 1

[0044] The difference between Comparative Example 1 and Example 1 is that: Comparative Example 1 does not undergo enzymolysis in step 2), that is, the angelica extract, the Polygonum multiflorum extract and the Platycladus orientalis extract are directly mixed and then fermented to obtain a plant fermentation composition. The remaining components and steps are the same as those in Example 1.

[0045] Comparative Example 2

[0046] The difference between Comparative Example 2 and Example 2 is that: Comparative Example 2 does not undergo enzymolysis in step 2), that is, the angelica extract, the Polygonum multiflorum extract and the Platycladus orientalis extract are directly mixed and then fermented to obtain a plant fermentation composition. The remaining components and steps are the same as those in Example 2.

[0047] Comparative Example 3

[0048] The difference between Comparative Example 3 and Comparative Example 1 is that the microorganism used in Comparative Example 3 is Bacillus subtilis. That is, compared with Example 1, Comparative Example 3 does not undergo an enzymatic hydrolysis step, and the inoculated co-fermentation microorganism is also replaced.

[0049] Comparative Example 4

[0050] The difference between Comparative Example 4 and Example 1 is that the microorganism used in Comparative Example 4 is Bacillus subtilis.

[0051] Comparative Example 5

[0052] The difference between Comparative Example 5 and Example 1 is that: Comparative Example 5 does not undergo fermentation in step 3), that is, the Angelica sinensis enzymatic hydrolysate and the Polygonum multiflorum and Platycladus orientalis leaf composite enzymatic hydrolysate are directly mixed to obtain the plant enzymatic hydrolysate composition.

[0053] Comparative Example 6

[0054] The difference between Comparative Example 6 and Example 1 is that: Comparative Example 6 does not undergo the enzymatic hydrolysis of step 2) and the fermentation of step 3), that is, the angelica extract, the Polygonum multiflorum extract and the Platycladus orientalis leaf extract are directly mixed to obtain the plant extract composition.

[0055] Comparative Example 7

[0056] The difference between Comparative Example 7 and Example 1 is that: in Comparative Example 7, no Angelica sinensis is added, that is, the plant fermentation composition only contains Polygonum multiflorum and Platycladus orientalis leaves.

[0057] Comparative Example 8

[0058] The difference between Comparative Example 8 and Example 1 is that: Comparative Example 8 does not add Polygonum multiflorum, that is, the plant fermentation composition only contains Angelica sinensis and Platycladus orientalis leaves.

[0059] Comparative Example 9

[0060] The difference between Comparative Example 9 and Example 1 is that: Comparative Example 8 does not add Platycladus orientalis leaves, that is, the plant fermentation composition only contains Angelica sinensis and Polygonum multiflorum.

[0061] Performance Testing

[0062] In the present invention, Example 1, Comparative Example 1, Comparative Example 5 and Comparative Example 6 are set as experimental group A; Example 1, Example 2 and Comparative Example 4 are set as experimental group B; Comparative Example 1, Comparative Example 2 and Comparative Example 3 are set as experimental group C; Example 1, Comparative Example 7, Comparative Example 8 and Comparative Example 9 are set as experimental group D.

[0063] Among them, experimental group A (Example 1, Comparative Example 1, Comparative Example 5 and Comparative Example 6) is to explore the effect of lacking one or two of enzymatic hydrolysis and fermentation on performance; experimental group B (Example 1, Example 2, Comparative Example 4) is to explore the effect of different types of microbial fermentation on performance. Experimental group C (Comparative Example 1, Comparative Example 2 and Comparative Example 3) is to explore the effect of different types of microbial fermentation on performance without enzymatic hydrolysis, so as to reduce the error caused by the enzymatic hydrolysis step. Experimental group B and experimental group C are combined for analysis to obtain the optimal microorganism. Experimental group D (Example 1, Comparative Example 7, Comparative Example 8 and Comparative Example 9) is to explore the effect of lacking one of Angelica sinensis, Platycladus orientalis and Polygonum multiflorum on the anti-hair loss and hair growth effect.

[0064] 1. Test of active ingredient content in each group (data see Table 1)

[0065] a. Microbial growth (OD 600 )

[0066] Method: The absorbance of the test sample at 600nm is used to characterize the growth of microorganisms.

[0067] Sample: the fermentation broth before concentration in Examples 1 to 2 and Comparative Examples 1 to 4.

[0068] b. Active ingredient content test

[0069] Methods: The contents of stilbene glycosides and flavonoids in the leaves of Platycladus orientalis were determined by HPLC, and the content of polysaccharides was determined by sulfuric acid-phenol method.

[0070] Samples: Examples 1 to 2, Comparative Examples 1 to 6

[0071] Table 1 Data on the content of active ingredients in each group

[0072]

[0073]

[0074] As can be seen from Table 1, in experimental group A, the active ingredients of the composition of Example 1 after enzymatic hydrolysis and fermentation are significantly higher than those of the same experimental group, while the contents of angelica polysaccharides and diphenylethylene glycosides in the composition without enzymatic hydrolysis in comparative example 1 have decreased, indicating that enzymatic hydrolysis can increase the contents of angelica polysaccharides and diphenylethylene glycosides, but has little effect on Platycladus orientalis leaf flavonoids; in comparative example 5, the content of angelica polysaccharides in the composition without fermentation is greatly reduced and the content of Platycladus orientalis leaf flavonoids is also slightly reduced, so the content of angelica polysaccharides is more affected by whether it is fermented, and fermentation has little effect on diphenylethylene glycosides. The composition without enzymatic hydrolysis and fermentation in comparative example 6 is simply a mixture of three plant compositions, and the contents of the three active ingredients are the lowest. Therefore, enzymatic hydrolysis and fermentation can increase the content of active ingredients to a greater extent, thereby improving the subsequent anti-hair loss and hair growth effect.

[0075] In experimental groups B and C, the active ingredients of the co-fermentation microorganism groups (Comparative Examples 3 and 4) inoculated with Bacillus subtilis were lower than those of the same experimental groups, especially the content of angelica polysaccharides and diphenylethylene glycosides, which were even close to the data of Comparative Example 6, indicating that Bacillus subtilis is not suitable for selection as a co-fermentation microorganism, so yeast and lactic acid bacteria are preferred as co-fermentation microorganisms.

[0076] 2. Stickiness and stability test of each group (data see Table 2)

[0077] a. Stability test method: Place the sample in a 48°C oven for 7 days, then take it out and return it to room temperature to observe the precipitation (compared with Comparative Example 6).

[0078] b. Stickiness test method: Apply the sample to the back of the hand and evaluate the stickiness of the sample by skin touch (compared with Comparative Example 6).

[0079] Samples: Examples 1 to 2, Comparative Examples 1 to 5

[0080] Table 2 Viscosity and stability of the compositions of each group

[0081] Group Sticky stability Example 1 decline No obvious precipitation Example 2 decline No obvious precipitation Comparative Example 1 decline Precipitation reduction (++) Comparative Example 2 decline Precipitation reduction (+) Comparative Example 3 constant constant Comparative Example 4 constant constant Comparative Example 5 constant constant

[0082] In Comparative Example 6, after the three plant extracts were mixed, there were a lot of precipitations after standing, indicating that the mixed stability of the extracts was poor. As shown in Table 2, in experimental group A, the viscosity of the composition of Example 1 after enzymatic hydrolysis and fermentation decreased while there was no obvious precipitation, while the viscosity of Comparative Example 1 after only fermentation remained unchanged and the precipitation decreased, and the viscosity and stability of Comparative Example 5 after only enzymatic hydrolysis remained unchanged, indicating that only enzymatic hydrolysis and only fermentation have little effect on the viscosity of the liquid, and the combination of enzymatic hydrolysis and fermentation can reduce the viscosity of the composition and significantly improve the stability.

[0083] In experimental group B and experimental group C, the viscosity and stability of the inoculated co-fermentation microorganism using Bacillus subtilis (Comparative Example 3 and Comparative Example 4) did not change compared with Comparative Example 6. Compared with Comparative Examples 1 and 2, the use of Bacillus subtilis fermentation will make the active substance agglomerate and unstable, so Bacillus subtilis is not suitable for selection as a co-fermentation microorganism. Compared with Comparative Example 2, Comparative Example 1 has less precipitation, indicating that the stability of lactic acid bacteria fermentation is higher, and lactic acid bacteria is more suitable for selection as a co-fermentation microorganism.

[0084] 3. 5α-reductase inhibition test of each group (data see Table 3)

[0085] Preparation of 5α-reductase: Five male SD rats were taken and humanely killed after being fasted overnight without water restriction. The prostate was removed and cut into pieces on an ice table to obtain the 5α-reductase extract, which was stored at low temperature for later use.

[0086] 5α-reductase activity determination: Take 0.5mL phosphate buffer, 0.1mL test solution, 200μL testosterone solution (300mg / L) and 200μL NADPH solution (0.8g / L), add 0.5mL 5α-reductase extract, react at 37℃ for 30min, add 2.5mL dichloromethane to stop the reaction and extract testosterone, then add 0.25mL propyl hydroxybenzoate (100mg / L) as internal standard, centrifuge at 3000r / min for 10min. Remove the upper aqueous phase, remove about 1mL organic phase and evaporate to dryness, dissolve the residue in 1.5mL methanol, take 10μL and use high performance liquid chromatography to determine the residual testosterone. Set up a 5α-reductase reaction tube and a 5α-reductase blank tube at the same time.

[0087] Test samples: Examples 1 to 2, Comparative Examples 1 to 9

[0088] 5α-reductase inhibition rate = (R 样品 -R 反应 ) / (R 空白 -R 反应 )×100%; where R is the ratio of the target peak area to the internal standard peak area. Positive control: 0.05 mg / L finasteride.

[0089] Table 3 Data of 5α-reductase inhibition rate in each group

[0090]

[0091]

[0092] The higher the 5α-reductase inhibition rate, the better the anti-hair loss effect. As can be seen from Table 4, Examples 1 to 2 have the best anti-hair loss effect. Among them, the effect of Example 1 is better, that is, the co-fermented microorganism is preferably lactic acid bacteria. The anti-hair loss effects of Comparative Examples 1 to 9 are all lower than those of Example 1, indicating that whether the fermentation composition has undergone enzymatic hydrolysis, fermentation, the selection of fermentation bacteria, and the combination of Chinese medicinal materials will affect its anti-hair loss effect. In combination, the Chinese medicinal fermentation composition scheme using lactic acid bacteria after water extraction of the three medicinal materials of Angelica sinensis, Polygonum multiflorum and Platycladus orientalis leaves, followed by enzymatic hydrolysis and fermentation, has the best anti-hair loss effect.

[0093] 4. Trial test of serum

[0094] The plant fermentation composition of Example 1 was added to the essence at a ratio of 5% to prepare the anti-hair loss essence. The formula of the scalp care essence is shown in Table 4.

[0095] Table 4 Scalp care essence formula

[0096]

[0097] Volunteer selection criteria: A total of 22 volunteers were recruited, 19 of whom completed the test, and 3 subjects dropped out midway. The final participants who completed the test were 16 females and 3 males, with an average age of 30.32±6.04 years. All completed subjects met the following inclusion criteria, exclusion criteria, and withdrawal (dropout) criteria:

[0098] Inclusion criteria: 1. 18-60 years old, healthy men or women; 2. Hair length between 5 and 40 cm; 3. Those who are troubled by excessive hair loss and mild hair thinning, and the hair loss count is greater than 10 strands according to the 60-combing method; 4. Those who have not undergone special hair treatments such as hair dyeing, perming, and styling in the past month; 5. Those who can understand the experimental process, voluntarily participate in the experiment and sign the written informed consent.

[0099] Exclusion criteria: 1. Pregnant or lactating women, or those who are planning to get pregnant in the near future; 2. Patients with severe androgenic alopecia, alopecia areata, inflammatory cicatricial alopecia or other scalp / hair diseases; 3. People with psychiatric or psychological diseases; or those with long-term sleep or emotion control disorders; 4. Those who have used cosmetics with anti-hair loss effects or other products with such effects or hair growth effects in the past 3 months; 5. Those who have taken or topically applied any drugs that affect hair growth in the past 6 months; 6. Those who have received hair transplant treatment; 7. Those with curly hair; 8. Those with highly sensitive constitutions; 9. Those who have participated in other clinical trials in the past 2 months; 10. Those who are considered unsuitable for participation in the trial by clinical evaluation.

[0100] Exit criteria: 1. The subject is lost to follow-up and actively requests to withdraw; 2. Poor compliance, failure to use the product on time and in the prescribed amount, and failure to return for follow-up as required; 3. Violation of the research protocol; 4. Serious adverse reactions occur during the entire test period.

[0101] Volunteers were selected to take the essence made from the plant fermentation composition of Example 1 once a day, apply it to the head, and massage for absorption, once a day. The volunteers' hair loss and hair density were tested after four weeks, eight weeks, and twelve weeks. The 60-times combing method was used to detect the change in the number of hair loss of the volunteers, and the dermatoscope was used to take photos and analyze the change in hair density.

[0102] Depend on Figures 1-2 It can be seen that after continuous use of the essence made from the plant fermentation composition of Example 1 for 12 weeks, hair loss was reduced by 48% and hair density was increased by 8% compared with before use, indicating that it has a good anti-hair loss and hair growth effect.

[0103] The above-mentioned embodiments are only preferred embodiments of the present invention and cannot be used to limit the scope of protection of the present invention. Any non-substantial changes and substitutions made by technicians in this field on the basis of the present invention shall fall within the scope of protection required by the present invention.

Claims

1. A method for preparing a plant fermentation composition for preventing hair loss and promoting hair growth, characterized in that: The following steps are involved: 1) After extracting angelica sinensis with water, adding amylase and treating at 40-60°C for 1-3h, inactivating the enzyme at 80-90°C for 20min-60min, filtering and collecting the supernatant to obtain angelica sinensis hydrolyzate; after extracting Polygonum multiflorum and Platycladus orientalis leaves with water, adding pectinase and treating at 40-60°C for 1-3h, inactivating the enzyme at 80-90°C for 20min-60min, filtering and collecting the supernatant to obtain Polygonum multiflorum and Platycladus orientalis leaf composite hydrolyzate; the mass ratio of one of angelica sinensis, Polygonum multiflorum and Platycladus orientalis leaves to water is 1:5-20; the mass ratio of angelica sinensis, Polygonum multiflorum and Platycladus orientalis leaves is 0.5-2:2-4:4-6; 2) combining the angelica enzymatic hydrolysate and the Polygonum multiflorum and Platycladus orientalis leaf composite enzymatic hydrolysate, inoculating microorganisms for co-fermentation, filtering and collecting the supernatant to obtain a plant fermentation composition; the inoculated co-fermentation microorganisms are lactic acid bacteria or yeast, and the fermentation conditions are: fermentation at 35-38° C. for 18-36 hours; wherein the inoculation amount of the co-fermentation microorganisms is 2-4% of the mass of the enzymatic hydrolysate.

2. The method for preparing the plant fermentation composition for preventing hair loss and promoting hair growth according to claim 1, characterized in that: In step 1), the temperature of water extraction of angelica, Polygonum multiflorum and Platycladus orientalis leaves is 85-90° C. and the time is 30-60 min.

3. The method for preparing the plant fermentation composition for preventing hair loss and promoting hair growth according to claim 1, characterized in that: In step 2), the Angelica sinensis hydrolysate and the Polygonum multiflorum and Platycladus orientalis leaf composite hydrolysate are mixed, and then inoculated with microorganisms for co-fermentation, the supernatant is filtered and concentrated to a crude drug concentration of 0.1-0.2 g / g to obtain a plant fermentation composition.

4. A plant fermentation composition for preventing hair loss and promoting hair growth, characterized in that: The plant fermented composition for preventing hair loss and promoting hair growth is prepared by the preparation method of the plant fermented composition for preventing hair loss and promoting hair growth according to any one of claims 1 to 3.

5. An application of a plant fermentation composition for preventing hair loss and promoting hair growth, characterized in that: The plant fermentation composition for preventing hair loss and promoting hair growth according to claim 4 is used to prepare an anti-hair loss and promoting hair growth product; wherein the mass percentage of the plant fermentation composition for preventing hair loss and promoting hair growth in the anti-hair loss and promoting hair growth product is 3-5%.

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