A preparation method and application of compound black red tincture

Abstract

The invention relates to the technical field of topical external medicines, and in particular to a preparation method and application of a compound black and red tincture. The compound black and red tincture comprises the following raw materials: psoralea corylifolia, drynaria rhizome, tribulus terrestris, red peony root, safflower, processed polygonum multiflorum, salvia miltiorrhiza, dried ginger, calcined natural copper, cassia twig, ligusticum chuanxiong and an extracting solution; the extracting solution is compounded with ethanol and an antioxidant in a specific proportion, and the extracting solution can be used in combination with the extraction method of the invention to extract active ingredients in various raw materials to the maximum extent in a short time, and the active ingredients can be stably present, thereby reducing the loss rate of the active ingredients; the compound black and red tincture prepared by the invention can be used for treating alopecia areata, treating white hair, treating vitiligo, inhibiting hair loss and promoting new hair growth, and has broad application prospects.

Description

Technical Field

[0001] The invention relates to the technical field of topical external medicines, and in particular to a preparation method of a compound black red tincture and application thereof. Background Art

[0002] Alopecia areata is a common chronic skin disease in dermatology, in which hair is not scarred and suddenly falls off in patches. The clinical manifestations are sudden patchy hair loss, thinning of the scalp at the place of hair loss, and mostly no subjective symptoms. Alopecia areata is closely related to alopecia totalis and alopecia universalis. Alopecia totalis refers to the loss of all hair. Alopecia universalis is based on alopecia totalis, and all axillary hair, eyebrows, eyelashes, and vellus hair fall off. The incidence of this disease is about 2%; it can be seen in all ages, genders, and races, and it is more common in young people. Studies have shown that more than 60% of patients are under 30 years old, among whom children are more susceptible to the disease than adults, female children develop the disease earlier, and boys develop the disease more severely. The pathogenesis of this disease is based on genetic background, coupled with the induction of the external environment, and the immune response of the local hair follicles. It is a T-cell-mediated autoimmune disease and a multi-gene complex disease. It is most related to the human leukocyte antigen class II region on human chromosome 6. The immune immunity of hair follicles is broken, the synergy of CD4+ and CD8+ T lymphocytes is unbalanced, and the participation of cytokines such as interleukin-1, vascular endothelial growth factor, hepatocyte growth factor, interferon-γ, and tumor necrosis factor-α causes hair follicles to be destroyed or enter a resting phase, causing hair loss and disease. Mental factors such as long-term stress, anxiety, depression, and insomnia can trigger this disease. After the disease occurs, the hair becomes oilier, the image is damaged, and the patient becomes more anxious. The two form a vicious cycle and aggravate the disease. It is also related to endocrine disorders. For example, patients with thyroid diseases are more likely to develop the disease.

[0003] In summary, alopecia areata currently has a high rate of hair loss and is not easy to cure, and the market demand for related drugs is large. Traditional treatments are mainly glucocorticoids, immunosuppressants, minoxidil, compound glycyrrhizic acid glycosides, and photochemotherapy, but side effects and drug resistance are difficult to solve. Among them, the side effects of glucocorticoids include: weight gain, thinning skin, susceptibility to infection and osteoporosis, etc.; the side effects of immunosuppressive drugs include: liver and kidney function damage, bone marrow suppression, infection, etc.; minoxidil has skin dandruff, burning pain, allergic rhinitis, in addition to minoxidil tincture has a "crazy shedding period", and some patients experience increased early hair loss after use; compound glycyrrhizic acid glycosides may cause high blood pressure and lead to side effects such as weight loss. The efficacy of these existing treatments for alopecia areata is still not clear enough, the effect is relatively single, and there are problems such as adverse reactions and inconvenience in use.

[0004] Therefore, according to the above-mentioned related technologies, it is urgent to develop a preparation method and application of a compound black red tincture. Summary of the invention

[0005] In view of this, the purpose of the present invention is to provide a preparation method of a compound black red tincture and its application, so as to provide a medicament for treating alopecia areata which has definite efficacy, no obvious adverse reactions and is easy to use.

[0006] Based on the above purpose, the present invention provides a preparation method of compound black red tincture and application thereof.

[0007] A compound black and red tincture comprises the following raw materials in parts by mass: 10-11 parts of psoralea corylifolia, 10-11 parts of drynaria rhizome, 20-22 parts of tribulus terrestris, 30-33 parts of red peony root, 6-6.6 parts of safflower, 12-13.2 parts of processed radix polygoni multiflori, 6-6.6 parts of salvia miltiorrhiza, 3-3.3 parts of dried ginger, 20-22 parts of calcined natural copper, 6-6.6 parts of cassia twig, 6-6.6 parts of ligusticum chuanxiong and 1300-1500 parts of extract.

[0008] Preferably, the extract is obtained by mixing ethanol and an antioxidant; The mass ratio of the ethanol to the antioxidant is 1300-1500:0.3-0.5.

[0009] Preferably, the ethanol is 55%-75% ethanol; The antioxidant is any one or two of rutin and hesperidin.

[0010] A method for preparing a compound black red tincture comprises the following steps: Step A1: Under a protective atmosphere, vacuum freeze-drying and freezing ball-milling are performed on Psoralea corylifolia, Drynaria fortunei, Tribulus terrestris, Paeonia lactiflora, Carthamus tinctorius, Radix Polygoni multiflori, Salvia miltiorrhiza, Dry ginger, calcined natural copper, Cinnamomum cassia twig, and Rhizoma Chuanxiong, respectively, and then added to the extract solution and soaked for 3-5 days to obtain an extract solution; Step A2: subjecting the extract to an ultrasonic-high pressure synergistic treatment, and then soaking the extract for 20-24 hours, subjecting the extract to an ultrasonic-high pressure synergistic treatment for a second time, and then soaking the extract for 20-24 hours before filtering to obtain the compound black red tincture.

[0011] Preferably, the protective atmosphere in step A1 is any one of nitrogen and argon.

[0012] Preferably, the vacuum freeze-drying treatment in step A1 is to first balance at -7--5°C for 30-40 minutes, then cool to -25--20°C and freeze for 8-10 hours, and then dry at -60--55°C and 5-8Pa for 22-25 hours; The temperature of the frozen ball milling treatment in step A1 is -30-25°C, the ball milling speed is 500-600r / min, and the ball milling time is 3-5min.

[0013] Preferably, the ultrasonic-high pressure synergistic treatment in step A2 is a step-by-step ultrasonic treatment, which is first ultrasonicated at 60-70 W and 40-45 kHz for 10-15 min, then ultrasonicated at 250-350 MPa for 3-5 min, and finally ultrasonicated at 150-170 W and 40-45 kHz for 5-10 min.

[0014] Preferably, the secondary ultrasound-high pressure synergistic treatment in step A2 is a step-by-step ultrasound treatment, which is first performed at 80-90W, 40-45kHz for 10-15min, followed by a high pressure treatment at 250-350MPa for 3-5min, and finally a secondary ultrasound at 150-170W, 40-45kHz for 5-10min.

[0015] Preferably, the pore size of the filter during the filtration in step A2 is 5-7.5 μm.

[0016] The invention discloses an application of a compound black and red tincture, wherein the compound black and red tincture is used for treating alopecia areata, treating white hair, treating vitiligo, inhibiting hair loss and promoting new hair growth.

[0017] The psoralea corylifolia used in the present invention has the effects of warming the kidney and invigorating yang, relieving asthma, warming the spleen and stopping diarrhea. It plays the role of warming yang and tonifying the kidney in the compound, laying the foundation for treating diseases such as alopecia areata. Drynaria can tonify the kidney and strengthen bones, heal injuries and relieve pain, and synergize with psoralea corylifolia to enhance the power of warming yang and tonifying the kidney, and promote the repair and growth of hair follicles; Tribulus terrestris has the effects of calming the liver and relieving depression, promoting blood circulation and dispelling wind, improving eyesight and relieving itching. In the compound, the combination of Tribulus terrestris and other blood-activating drugs can enhance the effect of promoting blood circulation and unblocking collaterals, improve local blood circulation, provide sufficient nutrition for hair follicles, and promote hair growth; Red peony root can clear away heat, cool blood, dissipate blood stasis and relieve pain, while safflower can promote blood circulation, unblock menstruation, dissipate blood stasis and relieve pain. The two are complementary to each other, enhancing the effect of promoting blood circulation and removing blood stasis, effectively relieving local congestion, improving blood supply around hair follicles and promoting hair growth. Prepared Polygonum multiflorum has the functions of nourishing liver and kidney, improving essence and blood, blackening hair, and strengthening tendons and bones. In the compound, prepared Polygonum multiflorum not only helps to treat gray hair, but also nourishes liver and kidney, provides sufficient nutrition for hair growth, and fundamentally improves hair quality and hair growth. Danshen can invigorate blood circulation, dispel blood stasis, relieve pain, clear the heart and relieve restlessness, cool blood and eliminate carbuncle, while Ganjiang can warm the middle and dispel cold, restore yang and dredge meridians, warm the lungs and eliminate phlegm. The combination of the two can further enhance the effect of warming yang and invigorating blood circulation, improve local blood circulation and promote hair growth. Calcined natural copper can dissipate blood stasis, relieve pain, and repair tendons and bones. In the compound, it can promote the repair and regeneration of damaged hair follicles and contribute to the normal growth of hair. Guizhi can induce sweating and relieve muscle pain, warm and dredge the meridians, and help yang and transform qi. Chuanxiong can invigorate blood circulation and qi, dispel wind and relieve pain. The combination of the two can warm and dredge the meridians, promote the circulation of qi and blood, and provide a good environment for hair growth. The ethanol in the extract is used as a solvent to effectively extract the active ingredients in each raw material, while the antioxidants (rutin from Sophora japonica, hesperidin, etc.) can make the extracted active ingredients stably exist in the compound black red tincture solution, prevent the oxidation and degradation of the active ingredients, and ensure the quality and efficacy of the product; Through the reasonable combination of the above-mentioned raw materials, the compound black and red tincture of the present invention can play multiple roles in warming yang and tonifying the kidney, promoting blood circulation and unblocking collaterals, relieving local congestion, treating white hair, treating vitiligo, inhibiting hair loss, promoting new hair growth, etc. while treating alopecia areata. The raw materials work together to produce unexpected technical effects.

[0018] Beneficial effects of the present invention: The invention provides a preparation method of a compound black and red tincture and an application thereof. The invention can make the extracted active ingredients stably exist in the compound black and red tincture liquid by mixing ethanol and an antioxidant in a specific ratio; The present invention uses a compound mixture of psoralea corylifolia, drynaria, tribulus terrestris, red peony root, safflower, processed Polygonum multiflorum, salvia miltiorrhiza, dried ginger, calcined natural copper, cassia twig, and ligusticum chuanxiong in a specific ratio, which can treat alopecia areata while playing the roles of warming yang and tonifying the kidney, promoting blood circulation and unblocking collaterals, relieving local stasis, treating white hair, treating vitiligo, inhibiting hair loss, and promoting the growth of new hair, and no adverse reactions are produced; According to the invention, psoralea corylifolia, drynaria rhizome, tribulus terrestris, red peony root, safflower, processed radix polygoni multiflori, salvia miltiorrhiza, dried ginger, calcined natural copper, cassia twig and ligusticum chuanxiong are first subjected to vacuum freeze drying treatment and freezing ball milling treatment in sequence under a protective atmosphere, and then are sequentially added into a self-made extracting solution of the invention for soaking, and then subjected to an ultrasonic-high pressure synergistic treatment, and then soaked for 20-24 hours, subjected to a second ultrasonic-high pressure synergistic treatment, and then filtered after soaking for 20-24 hours. The method can extract active ingredients in each raw material to the maximum extent in a short time, and the active ingredients are stably present, thereby reducing the loss rate of the active ingredients. Compared with the prior art, the compound black and red tincture prepared by the invention has definite curative effect, no obvious adverse reaction, is easy to use and has wider application prospects. DETAILED DESCRIPTION

[0019] In order to make the objectives, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with specific embodiments.

[0020] Embodiment 1: A method for preparing a compound black red tincture, comprising the following steps: S1: Mix 1300 g of 55% ethanol and 0.3 g of Rutin from Sophora japonica to obtain an extract; S2: Under nitrogen atmosphere, 10 g of Psoralea corylifolia, 10 g of Drynaria fortunei, 20 g of Tribulus terrestris, 30 g of Paeonia lactiflora, 6 g of Carthamus tinctorius, 12 g of Polygonum multiflorum processed, 6 g of Salvia miltiorrhiza, 3 g of Drynaria ginger, 20 g of calcined natural copper, 6 g of Cinnamomum cassia twig, and 6 g of Ligusticum chuanxiong were subjected to vacuum freeze-drying and freeze-milling treatment in sequence, and then added to 1300 g of extract solution in sequence and soaked for 3 days to obtain an extract solution; The vacuum freeze-drying treatment is to first balance at -7°C for 30 minutes, then cool to -25°C for 8 hours, and then dry at -60°C and 5Pa for 22 hours; The temperature during the freezing ball milling treatment is -30°C, the ball milling speed during the freezing ball milling treatment is 500r / min, and the ball milling time during the freezing ball milling treatment is 3min; S3: subjecting the extract to an ultrasonic-high pressure synergistic treatment, and then soaking for 20 hours, subjecting the extract to an ultrasonic-high pressure synergistic treatment for a second time, and then soaking for 20 hours and filtering with a filter with a pore size of 5 μm to obtain the compound black red tincture; The first ultrasound-high pressure coordinated treatment is a step-by-step ultrasound treatment, which is firstly performed at 60W and 40kHz for 10 minutes, then performed at 250MPa for 3 minutes, and finally performed at 150W and 40kHz for 5 minutes. The secondary ultrasound-high pressure synergistic treatment is a step-by-step ultrasound treatment, which is firstly performed at 80W and 40kHz for 10 minutes, then performed at 250MPa for 3 minutes, and finally performed secondary ultrasound at 150W and 40kHz for 5 minutes. Embodiment 2: A method for preparing a compound black red tincture, comprising the following steps: S1: Mix 1350 g of 60% ethanol and 0.35 g of hesperidin to obtain an extract; S2: Under argon atmosphere, 10.2 g of Psoralea corylifolia, 10.2 g of Drynaria fortunei, 20.4 g of Tribulus terrestris, 30.6 g of Paeonia lactiflora, 6.13 g of Carthamus tinctorius, 12.45 g of Radix Polygoni Multiflori, 6.12 g of Salvia miltiorrhiza, 3.06 g of Drynaria ginger, 20.4 g of calcined natural copper, 6.12 g of Cinnamomum cassia twig, and 6.12 g of Ligusticum chuanxiong were subjected to vacuum freeze-drying and freeze-milling treatment in sequence, and then added to 1350 g of extract solution and soaked for 3.5 days to obtain an extract solution; The vacuum freeze-drying process is to first balance at -6.5°C for 32 minutes, then cool to -24°C and freeze for 8.5 hours, and then dry at -58°C and 6Pa for 22.5 hours; The temperature during the freezing ball milling treatment is -29°C, the ball milling speed during the freezing ball milling treatment is 520r / min, and the ball milling time during the freezing ball milling treatment is 3.5min; S3: subjecting the extract to an ultrasonic-high pressure synergistic treatment, and then soaking for 21 hours, subjecting the extract to an ultrasonic-high pressure synergistic treatment for a second time, and then soaking for 21 hours and filtering with a filter with a pore size of 6 μm to obtain the compound black red tincture; The first ultrasound-high pressure coordinated treatment is a step-by-step ultrasound treatment, which is firstly performed at 62W and 40kHz for 11 minutes, then performed at 270MPa for 4 minutes, and finally performed at 155W and 40kHz for 6 minutes. The secondary ultrasound-high pressure synergistic treatment is a step-by-step ultrasound treatment, which is firstly performed at 82W and 40kHz for 11 minutes, then performed at 270MPa for 4 minutes, and finally performed secondary ultrasound at 155W and 40kHz for 6 minutes. Embodiment 3: A method for preparing a compound black red tincture, comprising the following steps: S1: Mix 1400 g of 65% ethanol and 0.4 g of antioxidant to obtain an extract; The antioxidant is a mixture of rutin and hesperidin; S2: Under nitrogen atmosphere, 10.5 g of Psoralea corylifolia, 10.5 g of Drynaria fortunei, 21 g of Tribulus terrestris, 31.51 g of Paeonia lactiflora, 6.3 g of Carthamus tinctorius, 12.6 g of Polygonum multiflorum processed, 6.3 g of Salvia miltiorrhiza, 3.16 g of Drynaria ginger, 21 g of calcined natural copper, 6.3 g of Cinnamomum cassia twig, and 6.31 g of Ligusticum chuanxiong were subjected to vacuum freeze-drying and freeze-milling treatment in sequence, and then added to 1400 g of extract solution and soaked for 4 days to obtain an extract solution; The vacuum freeze-drying process is to first balance at -6°C for 35 minutes, then cool to -23°C and freeze for 9 hours, and then dry at -58°C and 7Pa for 24 hours; The temperature during the freezing ball milling treatment is -28°C, the ball milling speed during the freezing ball milling treatment is 550r / min, and the ball milling time during the freezing ball milling treatment is 4min; S3: subjecting the extract to an ultrasonic-high pressure synergistic treatment, and then soaking for 22 hours, subjecting the extract to an ultrasonic-high pressure synergistic treatment for a second time, and then soaking for 22 hours and filtering with a filter with a pore size of 6.5 μm to obtain the compound black red tincture; The first ultrasound-high pressure coordinated treatment is a step-by-step ultrasound treatment, which is firstly performed at 65W and 43kHz for 13 minutes, then performed at 300MPa for 4 minutes, and finally performed at 160W and 43kHz for 8 minutes. The secondary ultrasound-high pressure synergistic treatment is a step-by-step ultrasound treatment, which is firstly performed at 85W and 43kHz for 13 minutes, then performed at 300MPa for 4 minutes, and finally performed secondary ultrasound at 160W and 43kHz for 7 minutes. Embodiment 4: A method for preparing a compound black red tincture, comprising the following steps: S1: Mix 1450 g of 70% ethanol and 0.45 g of rutin from sophora japonica to obtain an extract; S2: Under nitrogen atmosphere, 10.8 g of Psoralea corylifolia, 10.79 g of Drynaria fortunei, 21.5 g of Tribulus terrestris, 32.3 g of Red Peony Root, 6.5 g of Carthamus tinctorius, 12.9 g of Radix Polygoni Multiflori, 6.5 g of Salvia miltiorrhiza, 3.2 g of Drynaria ginger, 21.5 g of Calcined Copper Pyrite, 6.5 g of Cinnamomum cassia twig, and 6.5 g of Rhizoma Chuanxiong were subjected to vacuum freeze-drying and freeze-milling treatment in sequence, and then added to 1450 g of extract solution and soaked for 4.5 days to obtain an extract solution; The vacuum freeze-drying treatment is to first balance at -6.5°C for 39 minutes, then cool to -21°C and freeze for 9.5 hours, and then dry at -60--55°C and 7Pa for 24 hours; The temperature during the freezing ball milling treatment is -29°C, the ball milling speed during the freezing ball milling treatment is 580r / min, and the ball milling time during the freezing ball milling treatment is 4.5min; S3: subjecting the extract to an ultrasonic-high pressure synergistic treatment, and then soaking for 23 hours, subjecting the extract to an ultrasonic-high pressure synergistic treatment for a second time, and then soaking for 23 hours and filtering with a filter with a pore size of 7 μm to obtain the compound black red tincture; The first ultrasound-high pressure coordinated treatment is a step-by-step ultrasound treatment, which is firstly performed at 68W and 44kHz for 14 minutes, then performed at 330MPa for 4.5 minutes, and finally performed at 165W and 44kHz for 9 minutes. The secondary ultrasound-high pressure synergistic treatment is a step-by-step ultrasound treatment, which is firstly performed at 88W and 44kHz for 14 minutes, then performed at 340MPa for 4.5 minutes, and finally performed secondary ultrasound at 165W and 44kHz for 9 minutes. Embodiment 5: A method for preparing a compound black red tincture, comprising the following steps: S1: Mix 1500 g of 75% ethanol and 0.5 g of hesperidin to obtain an extract; S2: Under argon atmosphere, 11g of Psoralea corylifolia, 11g of Drynaria fortunei, 22g of Tribulus terrestris, 33g of Red Peony Root, 6.6g of Carthamus tinctorius, 13.2g of Radix Polygoni Multiflori, 6.6g of Salvia miltiorrhiza, 3.3g of Drynaria ginger, 22g of calcined natural copper, 6.6g of Cinnamomum cassia twig, and 6.6g of Rhizoma Chuanxiong were subjected to vacuum freeze-drying and freezing ball-milling in sequence, and then added to 1500g of extract solution in sequence and soaked for 5 days to obtain an extract solution; The vacuum freeze-drying treatment is to first balance at -5°C for 40 minutes, then cool to -20°C for 10 hours, and then dry at -55°C and 8Pa for 25 hours; The temperature during the freezing ball milling treatment is -25°C, the ball milling speed during the freezing ball milling treatment is 600r / min, and the ball milling time during the freezing ball milling treatment is 5min; S3: subjecting the extract to an ultrasonic-high pressure synergistic treatment, and then soaking the extract for 24 hours, subjecting the extract to an ultrasonic-high pressure synergistic treatment for a second time, and then soaking the extract for 24 hours and filtering the extract with a filter having a pore size of 7.5 μm to obtain the compound black red tincture; The first ultrasound-high pressure coordinated treatment is a step-by-step ultrasound treatment, which is firstly performed at 70W and 45kHz for 15 minutes, then performed at 350MPa for 5 minutes, and finally performed at 170W and 45kHz for 10 minutes. The secondary ultrasound-high pressure synergistic treatment is a step-by-step ultrasound treatment, which is firstly performed at 90W and 45kHz for 15 minutes, then performed at 350MPa for 5 minutes, and finally performed secondary ultrasound at 170W and 45kHz for 10 minutes. Comparative Example 1: Compared with Example 1, this comparative example did not add an antioxidant during the preparation of the extract, and the remaining steps and parameters were the same, which will not be repeated in this comparative example, and finally obtained the compound black red tincture; Comparative Example 2: Compared with Example 1, this comparative example did not use a protective atmosphere in the preparation process of the extract, and the remaining steps and parameters were the same, and this comparative example will not be repeated, and finally the compound black red tincture was obtained; Comparative Example 3: Compared with Example 1, this comparative example only replaces the "vacuum freeze-drying treatment" used in the preparation process of the extract with "drying treatment at 65°C", and the remaining steps and parameters are the same, which will not be repeated in this comparative example, and finally obtains the compound black red tincture; Comparative Example 4: Compared with Example 1, this comparative example only replaces the "vacuum freeze drying treatment and frozen ball milling treatment" used in the preparation process of the extract with "drying treatment at 65°C and crushing treatment at room temperature", and the remaining steps and parameters are the same, which will not be repeated in this comparative example, and finally obtains the compound black red tincture; Comparative Example 5: Compared with Example 1, the second ultrasonic-high pressure synergistic treatment was not performed in the preparation process of the compound black and red tincture in this comparative example, and the remaining steps and parameters were the same, which will not be repeated in this comparative example, and finally the compound black and red tincture was obtained; Comparative Example 6: S1: Mix 1300 g of 55% ethanol and 0.3 g of antioxidant to obtain an extract; The antioxidant is rutin; S2: Under nitrogen atmosphere, 10g of Psoralea corylifolia, 10g of Drynaria fortunei, 20g of Tribulus terrestris, 30g of Red Peony Root, 6g of Carthamus tinctorius, 12g of Radix Polygoni Multiflori, 6g of Salvia miltiorrhiza, 3g of Drynaria ginger, 20g of calcined natural copper, 6g of Cinnamomum cassia twig, and 6g of Rhizoma Chuanxiong were subjected to vacuum freeze-drying and freeze-ball milling in sequence, and then added to 1300g of extract solution in sequence and soaked for 7 days to obtain compound Wuhong tincture; The vacuum freeze-drying treatment is to first balance at -7°C for 30 minutes, then cool to -25°C for 8 hours, and then dry at -60°C and 5Pa for 22 hours; The temperature during the freezing ball milling treatment is -30°C, the ball milling speed during the freezing ball milling treatment is 500r / min, and the ball milling time during the freezing ball milling treatment is 3min; Comparative Example 7: S1: 10g of Psoralea corylifolia, 10g of Drynaria fortunei, 20g of Tribulus terrestris, 30g of Paeonia lactiflora, 6g of Carthamus tinctorius, 12g of Radix Polygoni multiflori, 6g of Salvia miltiorrhiza, 3g of Drynaria ginger, 20g of calcined natural copper, 6g of Cinnamomum cassia twig and 6g of Rhizoma Chuanxiong were dried at 65°C and crushed at room temperature, then added into 1300g of 55% ethanol and soaked for 7 days to obtain the compound Wuhong tincture.

[0021] Performance Testing: 1. Animal Testing Determination of new hair length: Kunming mice, half male and half female, a total of 130 mice, were randomly divided into 13 groups, 10 mice in each group, and the hair on the back of the mice was removed using a depilatory dose (the depilatory area was 3 cm × 4 cm). The compound black red tincture prepared in Examples 1-5 and Comparative Examples 1-7 was applied to the back of each group of mice, and the control group was applied with an equal amount of normal saline, once a day, for three consecutive weeks. The 10 longest hairs were taken at the same position of the depilatory area, the length was measured, the average length was calculated, and the results were statistically recorded in Table 1.

[0022] Determination of tyrosinase activity activation rate: The compound black red tincture prepared in Examples 1-5 and Comparative Examples 1-10 was mixed with a PBS solution to prepare a sample solution with a concentration of 0.5 mg / mL; then A, B, C, and D solutions were prepared according to the following method: A was 100 μL PBS + 50 μL tyrosinase solution (500 μ / mL); B was 100 μL PBS + 50 μL PBS; C was 100 μL sample solution + 50 μL tyrosinase solution (500 μ / mL); D was 100 μL sample solution + 50 μL PBS. Solutions A, B, C, and D were placed in a constant temperature water bath at 37°C for 5 minutes, 50 μL levodopa was added, and the temperature was continued for 2 minutes, the OD value (475 nm) was detected, and the activation rate of tyrosinase activity was calculated, wherein the activation rate = [(CD)-(AB)] / (AB) × 100%, and the results were statistically recorded in Table 1.

[0023] Table 1

[0024] Data Analysis: As can be seen from Table 1, the compound black-red tincture prepared by the present invention has better ability to promote new hair growth and tyrosinase activation (i.e., ability to treat gray hair); 2. Clinical Trials Clinical effects: White hair turning black rate: A group of 100 people (healthy men or women) participated in the white hair treatment test, aged 20-45 years old, with a total amount of white hair ≤ 1000 strands, with 28 days as a cycle, and every 3 cycles as a course of treatment. According to this method, the average white hair blackening rate of each group after one course of treatment using the compound black-red tincture prepared by Examples 1 to 5 of the present invention and Comparative Examples 1 to 7 was determined. The white hair blackening rate of each group is shown in Table 2; Hair loss cure rate: A group of 100 people (healthy men or women) aged 20-55 years old participated in the test to inhibit hair loss. The treatment was for seborrheic alopecia, alopecia areata, alopecia universalis, neurotic alopecia, postpartum alopecia and other alopecia. The effective rate was calculated by growing new hair. 28 days was a cycle, and every 3 cycles was a course of treatment. The cure rate of the compound black and red tincture prepared by Examples 1 to 5 of the present invention and Comparative Examples 1 to 7 after one course of treatment was determined according to this method. The cure rates of each group are shown in Table 2. Vitiligo cure rate: A group of 100 people (healthy men or women) aged 20-55 years old participated in the test of vitiligo treatment. The effective rate was calculated based on the absence of vitiligo symptoms. 28 days was a cycle and 3 cycles were a course of treatment. The cure rate of the compound black red tincture prepared by Examples 1 to 5 of the present invention and Comparative Examples 1 to 7 after one course of treatment was determined according to this method. The cure rates of each group are shown in Table 2. Directions: 1. When treating hair loss, the dosage is about 3-9 mL each time, depending on the degree of hair loss. Use twice a day (once in the morning and once in the evening, with an interval of about 8 hours), 28 days as a cycle, and 3 cycles as a course of treatment.

[0025] 2. When treating gray hair, randomly pull out the gray hair and then spray the tincture produced in Example 1 on the hair pulling site, using about 1 mL each time. Twice a day (preferably once in the morning and once in the evening, with an interval of about 8 hours), every 28 days is a cycle, and 3 cycles are a course of treatment. Note: Plucking out gray hair is to empty the hair follicles and allow the hair follicles to absorb the applied drug nutrients to the maximum extent. The targeted effect of the drug can directly open up the hair follicle nutrition channel. With the help of external force to supplement and help synthesize melanin particles, the empty hair follicles that have been pulled out of the gray hair can restore new black hair. From a physiological point of view, the melanocytes that secrete melanin are located at the bottom of the hair follicles. Plucking out gray hair is more conducive to the absorption and efficacy of active ingredients.

[0026] 3. When treating vitiligo, apply it evenly to the affected area according to the size of the affected area, twice a day (preferably once in the morning and once in the evening, with an interval of about 8 hours). Each 28 days is a cycle, and 3 cycles are a course of treatment.

[0027] Table 2 below shows the effects after one course of treatment, with 28 days as one cycle and 3 cycles as one course of treatment.

[0028] Table 2

[0029] Analysis of the clinical trial process and results shows that the compound black red tincture prepared by the present invention is not only easy to use, but also can have an excellent therapeutic effect on symptoms such as gray hair, hair loss, vitiligo, etc. in a short period of time.

[0030] 3. Clinical trial application Case 1, medical record number 103507117, female, 34 years old, alopecia areata, hair loss area on the top of the head, hair loss area ≥ 25% evenly distributed, hair loss time about 2 years, using the hair growth and hair blackening liquid embodiment 4 of the present invention. When treating hair loss, routinely disinfect the hair loss area, use a disinfected seven-star plum blossom needle to pierce, pierce evenly from the center of the lesion to the surrounding area, with local slight bleeding as the degree, once every other day, after piercing, apply the tincture produced by embodiment 4 externally, with the skin of the hair loss area completely moistened as the standard, and the dosage is 3-5ml according to the degree of hair loss. 2 times a day (preferably once in the morning and evening, with an interval of about 8 hours), 28 days as a cycle, and three cycles as a course of treatment. After three courses of treatment, the effect is significant, and the new hair covers more than 98% of the head.

[0031] Case 2, medical record number 576837, male, 36 years old, androgenic alopecia, hair loss area head and forehead, hair loss area ≥ 32% evenly distributed, hair loss time about 3 years, use the hair growth and hair blackening liquid embodiment 4 of the present invention. When treating hair loss, routinely disinfect the hair loss area, use the disinfected seven-star plum blossom needle to pierce, pierce evenly from the center of the lesion to the surrounding area, with local slight bleeding as the degree, once every other day, after piercing, apply the tincture produced by embodiment 4 externally, with the skin of the hair loss area completely moistened as the standard, and the dosage is 4-5ml according to the degree of hair loss. 2 times a day (preferably once in the morning and evening, with an interval of about 8 hours), 28 days as a cycle, and three cycles as a course of treatment. After three courses of treatment, the effect is significant, and the new hair covers more than 95% of the head.

[0032] Case 3, medical record number 103507098, male, 42 years old, androgenic alopecia, hair loss area top of the head, hair loss area ≥ 38% evenly distributed, hair loss time about 3 years, using the hair growth and darkening liquid embodiment 4 of the present invention. When treating hair loss, routinely disinfect the hair loss area, use a sterilized seven-star plum blossom needle to pierce, pierce evenly from the center of the lesion to the surrounding area, with local slight bleeding as the degree, once every other day, after piercing, apply the tincture produced by embodiment 4 externally, with the skin of the hair loss area completely moistened as the standard, and the dosage is 8-9ml according to the degree of hair loss. 2 times a day (preferably once in the morning and evening, with an interval of about 8 hours), 28 days as a cycle, and three cycles as a course of treatment. After three courses of treatment, the effect is significant, and the new hair covers more than 97% of the head.

[0033] Case 4, medical record number 103566443, female, 28 years old, alopecia areata, hair loss area on the top of the head, hair loss area ≥ 20% evenly distributed, hair loss time about 1 year, using the hair growth and hair blackening liquid embodiment 4 of the present invention. When treating hair loss, routinely disinfect the hair loss area, use a disinfected seven-star plum blossom needle to pierce, pierce evenly from the center of the lesion to the surrounding area, with local slight bleeding as the degree, once every other day, after piercing, apply the tincture produced by embodiment 4 externally, with the skin of the hair loss area completely moistened as the standard, and the dosage is 3-4ml according to the degree of hair loss. 2 times a day (preferably once in the morning and evening, with an interval of about 8 hours), 28 days as a cycle, and three cycles as a course of treatment. After three courses of treatment, the effect is significant, and the new hair covers more than 98% of the head.

[0034] Case 5, medical record number 103566817, male, 37 years old, androgenic alopecia, hair loss area top of the head, hair loss area ≥ 28% evenly distributed, hair loss time about 1 year, use the hair growth and hair blackening liquid embodiment 4 of the present invention. When treating hair loss, routinely disinfect the hair loss area, use the disinfected seven-star plum blossom needle to pierce, pierce evenly from the center of the lesion to the surrounding area, with local slight bleeding as the degree, once every other day, after piercing, apply the tincture produced by embodiment 4 externally, with the skin of the hair loss area completely moistened as the standard, and the dosage is 3-5ml according to the degree of hair loss. 2 times a day (preferably once in the morning and evening, with an interval of about 8 hours), 28 days as a cycle, and three cycles as a course of treatment. After three courses of treatment, the effect is significant, and the new hair covers more than 97% of the head.

[0035] Case 6, medical record number 103565147, female, 30 years old, androgenic alopecia, hair loss area top of the head, hair loss area ≥ 36% evenly distributed, hair loss time about 2 years, using the hair growth and dark hair liquid embodiment 4 of the present invention. When treating hair loss, routinely disinfect the hair loss area, use a disinfected seven-star plum blossom needle to pierce, pierce evenly from the center of the lesion to the surrounding area, with local slight bleeding as the degree, once every other day, after piercing, apply the tincture produced by embodiment 4 externally, with the skin of the hair loss area completely moistened as the standard, and the dosage is 5-6ml according to the degree of hair loss. Twice a day (preferably once in the morning and evening, with an interval of about 8 hours), 28 days as a cycle, and three cycles as a course of treatment. After three courses of treatment, the effect is significant, and the new hair covers more than 98% of the head.

[0036] Those skilled in the art should understand that the discussion of any of the above embodiments is merely illustrative and is not intended to imply that the scope of the present invention is limited to these examples. Under the concept of the present invention, the technical features in the above embodiments or different embodiments may be combined, the steps may be implemented in any order, and there are many other variations of the different aspects of the present invention as described above, which are not provided in detail for the sake of simplicity.

[0037] The present invention is intended to cover all such substitutions, modifications and variations that fall within the broad scope of the appended claims. Therefore, any omissions, modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A compound black red tincture, characterized in that: The invention comprises the following raw materials in parts by weight: 10-11 parts of psoralea corylifolia, 10-11 parts of drynaria rhizome, 20-22 parts of tribulus terrestris, 30-33 parts of red peony root, 6-6.6 parts of safflower, 12-13.2 parts of processed Polygonum multiflorum, 6-6.6 parts of salvia miltiorrhiza, 3-3.3 parts of dried ginger, 20-22 parts of calcined natural copper, 6-6.6 parts of cassia twig, 6-6.6 parts of ligusticum chuanxiong and 1300-1500 parts of extract.

2. The compound black red tincture according to claim 1, characterized in that The extract is obtained by mixing ethanol and an antioxidant; The mass ratio of the ethanol to the antioxidant is 1300-1500:0.3-0.

5.

3. The compound black red tincture according to claim 2, characterized in that The ethanol is 55%-75% ethanol; The antioxidant is any one or two of rutin and hesperidin.

4. A method for preparing a compound black red tincture, characterized in that: The following steps are involved: Step A1: Under a protective atmosphere, vacuum freeze-drying and freezing ball-milling are performed on Psoralea corylifolia, Drynaria fortunei, Tribulus terrestris, Paeonia lactiflora, Carthamus tinctorius, Radix Polygoni multiflori, Salvia miltiorrhiza, Dry ginger, calcined natural copper, Cinnamomum cassia twig, and Rhizoma Chuanxiong, respectively, and then added to the extract solution and soaked for 3-5 days to obtain an extract solution; Step A2: subjecting the extract to an ultrasonic-high pressure synergistic treatment, and then soaking the extract for 20-24 hours, subjecting the extract to an ultrasonic-high pressure synergistic treatment for a second time, and then soaking the extract for 20-24 hours before filtering to obtain the compound black red tincture.

5. The method for preparing the compound black red tincture according to claim 4, characterized in that: The protective atmosphere in step A1 is any one of nitrogen and argon.

6. The method for preparing the compound black red tincture according to claim 4, characterized in that: The vacuum freeze-drying treatment in step A1 is to first balance at -7--5°C for 30-40 minutes, then cool to -25--20°C and freeze for 8-10 hours, and then dry at -60--55°C and 5-8Pa for 22-25 hours; The temperature of the frozen ball milling treatment in step A1 is -30-25°C, the ball milling speed is 500-600r / min, and the ball milling time is 3-5min.

7. The method for preparing the compound black red tincture according to claim 4, characterized in that: The ultrasonic-high pressure synergistic treatment in step A2 is a step-by-step ultrasonic treatment, which is firstly ultrasonicated at 60-70W and 40-45kHz for 10-15min, then subjected to high pressure treatment at 250-350MPa for 3-5min, and finally subjected to ultrasonication at 150-170W and 40-45kHz for 5-10min.

8. The method for preparing the compound black red tincture according to claim 4, characterized in that: The secondary ultrasound-high pressure synergistic treatment in step A2 is a step-by-step ultrasound treatment, which is first performed at 80-90W, 40-45kHz for 10-15min, then performed at 250-350MPa for 3-5min, and finally performed at 150-170W, 40-45kHz for 5-10min.

9. The method for preparing the compound black red tincture according to claim 4, characterized in that: The pore size of the filter during the filtration in step A2 is 5-7.5 μm.

10. An application of compound black red tincture, characterized in that: The compound black and red tincture is used in preparing products for treating alopecia areata, treating white hair, treating vitiligo, inhibiting hair loss and promoting new hair growth.

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