KR20200136964A - Method for producing a composition for treating hair loss - Google Patents

Abstract

The manufacturing method of the composition for treating hair loss relates to three compositions for treating hair loss, and relates to the field of daily chemical products. The composition is intended to solve a technical problem in which the hair loss phenomenon of humans becomes more serious and the age group becomes younger. The method for preparing the composition includes the step of separating the polysaccharide of wood ear mushrooms (1); (2) preparing a fermentation substrate; And it includes the step (3) of obtaining a composition for treating hair loss by inoculating the fermentation substrate with yeast fungus, hairy fungus or Bacillus bacillus. The composition for treating hair loss has a remarkable promoting effect on hair follicle growth and a remarkable improvement effect on seborrheic hair loss.

Description

Method for producing a composition for treating hair loss

The present invention relates to a method for preparing various compositions, and to the field of daily chemical products.

As the rhythm of life increases, work stress increases, and food and lifestyle changes have taken place, the incidence of hair loss among Chinese young men has increased more than 10 times compared to 20 years ago. At the same time, the incidence of hair loss among female residents is also increasing year by year. However, the recognition rate that hair loss is considered a disease and needs to be visited and diagnosed is still low. The main causes of hair loss are genetic factors, physiological malfunction, strong stress from work and learning, mental tension, drug reactions, radiation therapy, environmental pollution, and malnutrition.

However, recent studies have found that hair loss is associated with an organism's intestinal microflora imbalance.Intestinal bacterial flora disorder increases Lactobacillus murnus, resulting in a deficiency of some substances or enzymes, which must be in the intestinal microbial secretion process. It is very likely to go through. The human gut flora is a complex colony. The total number of intestinal microbes in healthy adults is about 1 to 1.5 kg, and there are over 1000 species, and the number of intestinal microbes is 10 times the number of human cells, making it a "human transparent super organ". Intestinal flora is one of the important causes of metabolic abnormalities such as obesity, diabetes, tumors, hair loss, etc. Thus, this patent is the composition of the present invention. It is intended to supplement the type of intestinal flora by oral administration and to improve the intestinal flora.

An object of the present invention is to provide a method for preparing a composition for treating hair loss in order to solve the technical problem of human hair loss by improving the intestinal flora.

The first method of preparing a composition for treating hair loss,

Wash the black ear mushrooms thoroughly, remove impurities, and screen with 40 mesh after crushing to prepare black ear mushroom powder, and add black ear mushroom powder in distilled water according to the 140:1 mass ratio of black ear mushroom powder and distilled water. , Ultrasound is performed for 15 minutes under the condition that the ultrasonic power is 500 W, and after leaching in a water bath for 3 hours, the extract is centrifuged at 4500 r/min for 10 minutes to remove impurities, and then the extract is used as black ear mushrooms. Prepared as a material solution with a polysaccharide concentration of 4 g/L and a pH value of 5.6, and then, ultrafiltration is performed under conditions of a temperature of 40 °C and a pressure of 0.17 bar, and a blocking solution of a wood ear mushroom polysaccharide with a molecular blocking amount of 300 kDa. Membrane separation step (1) of the polysaccharide of wood ear mushrooms to obtain And

Select and weigh and clean yellow beans with large granules and uniform size, mix with water at a volume ratio of 1:3, soak for 3 hours at 30°C, boil and cool cooked ingredients to 30°C to 40°C Then, inoculate the polysaccharide blocking solution of wood ear mushrooms with a molecular blocking amount of 300 kDa and the yeast fungus inoculum at a mass ratio of 1:3, maintain room temperature at 28 ℃, and perform koji making for 20 to 24 hours. Perform turning koji, imperial for 28 hours to perform secondary soup overturning, finished koji after imperial for 34 hours, wash the soup after departure, and steam for 12 hours Into, and heat-retaining fermentation in a constant temperature room at 28° C. for 30 days or more, and then taken out and dried to obtain a 300 kDa blocking liquid Douchi fermented product.

The second method of preparing a composition for treating hair loss,

Wash the black ear mushrooms thoroughly, remove impurities, and screen with 40 mesh after crushing to prepare black ear mushroom powder, and add black ear mushroom powder in distilled water according to the 140:1 mass ratio of black ear mushroom powder and distilled water. , Ultrasound is performed for 15 minutes under the condition that the ultrasonic power is 500 W, and after leaching in a water bath for 3 hours, the extract is centrifuged at 4500 r/min for 10 minutes to remove impurities, and then the extract is used as black ear mushrooms. Prepared as a material solution with a polysaccharide concentration of 4 g/L and a pH value of 5.6, and then, ultrafiltration is performed under conditions of a temperature of 40 °C and a pressure of 0.17 bar, and a blocking solution of a wood ear mushroom polysaccharide with a molecular blocking amount of 300 kDa. Membrane separation step (1) of the polysaccharide of wood ear mushrooms to obtain

Put the hairy fungus species in sterile physiological saline, absorb the bacteria solution with a pipette and inoculate it into the bran medium, incubate at 20°C for 96 hours, and add 500 mL of sterile physiological saline to the cultured mother starter culture bottle. Put in, shake sufficiently, filter with double gauze, wash the filtered residue again with 500 mL of sterile physiological saline and filter, mix the two filtrates to prepare a hairy mildew spore suspension, and prepare the hairy fungus spore suspension 4 Putting it in a refrigerator at ℃ (2);

Select a yellow bean with a large granule and a uniform size, weigh and wash thoroughly, mix with water at a volume ratio of 1:3, soak at 30°C for 3 hours, drain the water, and then use a high-temperature autoclave, at 121°C. Steaming for 30 minutes, and then cooling (3), and

According to the mass ratio of 1:3, a molecular blocking amount of 300 kDa was inoculated with a polysaccharide blocking solution and a fungal spore suspension, incubated for 48 hours at a temperature of 25 ℃, and then subjected to post-ripening treatment for 48 hours, And a mixed inoculation step (4) to obtain a mushroom polysaccharide 30000 Da duchi fermented product.

The third method of preparing a composition for treating hair loss,

Wash the black ear mushrooms thoroughly, remove impurities, and screen with 40 mesh after crushing to prepare black ear mushroom powder, and add black ear mushroom powder in distilled water according to the 140:1 mass ratio of black ear mushroom powder and distilled water. , Ultrasound is performed for 15 minutes under the condition that the ultrasonic power is 500 W, and after leaching in a water bath for 3 hours, the extract is centrifuged at 4500 r/min for 10 minutes to remove impurities, and then the extract is used as black ear mushrooms. Prepared as a material solution with a polysaccharide concentration of 4 g/L and a pH value of 5.6, and then, ultrafiltration is performed under conditions of a temperature of 40 °C and a pressure of 0.17 bar, and a blocking solution of a wood ear mushroom polysaccharide with a molecular blocking amount of 300 kDa. Membrane separation step (1) of the polysaccharide of wood ear mushrooms to obtain

Steps to obtain a fermentation substrate by selecting and weighing and washing yellow bean of uniform size with rich granules, mixing with water at a volume ratio of 1:3, immersing in water for 10 to 18 hours, and sterilizing at 120 ℃ (2 ); And

The fermentation substrate is inoculated into the complex solution until the concentration of the complex solution in the fermentation substrate becomes 2 ml/L, and the complex solution is mixed with the Bacillus inoculum in a mass ratio of 1:3 with a 300 kDa blocking solution of a wood ear mushroom polysaccharide. , Shaking evenly, and incubating at 4 to 8° C. for 72 to 120 hours, and obtaining a Bacillus balance-neck ear mushroom polysaccharide 300 kDa blocking liquid Douchi fermentation product (3).

Duchi (fermented yellow bean) contains abundant nutrient substances, mainly proteins, free amino acids, essential fatty acids, soluble sugars, mineral salts, vitamins, and the like. In addition, Tochigi further contains physiologically active substances such as soybean isoflavone, soybean saponin, brown pigment, porpoise fibrinolytic enzyme, β-glucosidase, and antibiotics. Some of these active substances in bean curd are derived from raw soybeans, and part of them are derived from various microorganisms participating in the fermentation process and complex biochemical reactions such as decomposition and recombination of organic substances in raw soybeans.

Polysaccharides contained in black ear mushrooms have functions such as anti-ulcer, aging delay, blood lipid reduction, anti-hepatitis, anti-mutagenic, anti-tumor, protein and nucleic acid enhancement, and promotion of organism immunity, and block polysaccharides of wood ear mushrooms by membrane separation. Separated and fermented like bean curd, the hair firming effect is remarkable.

The features of the present invention are as follows.

1. Existing hair growth is to promote microcirculation of the scalp by performing an epidermal massage with ginger, cypress, etc., but if the fermented product of the present invention is administered orally, it is possible to restore the structure of the intestinal flora and restore micro-ecological equilibrium. . Controlled through the intestinal microflora administration to achieve the purpose of firming hair, this field is unexplored.

2. The composition of the present invention showed an extremely excellent hair firming effect in animal experiments, solved the hair loss problem on a large scale for the future, and found a new route.

3. The composition for treating hair loss of the present invention has a remarkable promoting effect on hair follicle growth and a remarkable improvement effect on seborrheic hair loss.

The technical solution of the present invention is not limited to the following specific embodiments, and further includes any combination between each specific embodiment.

In specific embodiment 1, the method for producing a composition for treating hair loss,

Wash the black ear mushrooms thoroughly, remove impurities, and screen with 40 mesh after crushing to prepare black ear mushroom powder, and add black ear mushroom powder in distilled water according to the 140:1 mass ratio of black ear mushroom powder and distilled water. , Ultrasound is performed for 15 minutes under the condition that the ultrasonic power is 500 W, and after leaching in a water bath for 3 hours, the extract is centrifuged at 4500 r/min for 10 minutes to remove impurities, and then the extract is used as black ear mushrooms. Prepared as a material solution with a polysaccharide concentration of 4 g/L and a pH value of 5.6, and then, ultrafiltration is performed under conditions of a temperature of 40 °C and a pressure of 0.17 bar, and a blocking solution of a wood ear mushroom polysaccharide with a molecular blocking amount of 300 kDa. Membrane separation step (1) of the polysaccharide of wood ear mushrooms to obtain And

Select a yellow bean with a large granule and a uniform size, weigh and wash thoroughly, mix with water at a volume ratio of 1:3, soak for 3 hours at 30°C (water content 45%-55%), boil and cook. After cooling the material to 30 ℃ to 40 ℃, inoculate the polysaccharide blocking solution and yeast fungus inoculum with a molecular blocking amount of 300 kDa at a mass ratio of 1:3, and maintain the room temperature at 28 ℃, 20 ~ 24 hours During the empire (koji making) to perform the first soup turning (turning koji), the empire for 28 hours to perform the second soup reversal, after 34 hours imperial (finished koji), and after leaving the country washing the soup And, put in a container by steaming for 12 hours, heat-retaining fermentation in a constant temperature room at 28°C for more than 30 days, and then take out and dry to obtain a fermented product of 300 kDa blocking solution of koji mold-tree ear mushroom polysaccharide. Include.

In step (1) of this embodiment, the leaching temperature in the water bath is 70°C to 85°C.

In step (1) of this embodiment, distilled water is added to the centrifugation supernatant, and the black ear mushroom polysaccharide in the supernatant is diluted to 4 g/L.

In step (2) of this embodiment, the said ripe material is boiled yellow bean.

In step (2) of the present embodiment, per 200 g of boiled and cooked yellow bean, a total of 3 ml of wood ear mushroom polysaccharide blocking solution and yeast fungus inoculum were inoculated, and the concentration of yeast fungus solution spores was 1.0 Х10 ⁷ ~ 1.0 Х10 ⁸ pcs/ml to be.

In the step (2) of the present embodiment, "to raise steam" means processing in a closed incubator at a temperature of 30±2°C.

In the specific embodiment 2, the difference between this embodiment and the specific embodiment 1 is that the moisture content is 50% in step (2). The rest are the same as in the specific embodiment 1.

In the specific embodiment 3, the difference between the present embodiment and the specific embodiment 1 or specific embodiment 2 is that the material cooked in step (2) is cooled to 35°C. The rest are the same as in the specific embodiment 1 or the specific embodiment 2.

In the specific embodiment 4, the difference between this specific embodiment and the specific embodiments 1 to 3 is that in step (2), the primary station is reversed by impinging for 22 hours. The rest are the same as in the specific embodiment 1 to the specific embodiment 3.

In specific embodiment 5, the manufacturing method of the composition for hair loss treatment of this specific embodiment,

Wash the black ear mushrooms thoroughly, remove impurities, and screen with 40 mesh after crushing to prepare black ear mushroom powder, and add black ear mushroom powder in distilled water according to the 140:1 mass ratio of black ear mushroom powder and distilled water. , Ultrasound is performed for 15 minutes under the condition that the ultrasonic power is 500 W, and after leaching in a water bath for 3 hours, the extract is centrifuged at 4500 r/min for 10 minutes to remove impurities, and then the extract is used as black ear mushrooms. Prepared as a material solution with a polysaccharide concentration of 4 g/L and a pH value of 5.6, and then, ultrafiltration is performed under conditions of a temperature of 40 °C and a pressure of 0.17 bar, and a blocking solution of a wood ear mushroom polysaccharide with a molecular blocking amount of 300 kDa. Membrane separation step (1) of the polysaccharide of wood ear mushrooms to obtain

Put the hairy fungus species in sterile physiological saline, absorb the bacteria solution with a pipette and inoculate it into the bran medium, incubate for 96 hours at 20°C (after the bran inside the triangular flask grows full of mycelium and gray spores, take out and prepare for use) , 500 mL of sterile physiological saline was added to the cultured mother starter culture bottle and sufficiently shaken, filtered with double gauze, and the filtered residue was washed once again with 500 mL of sterile physiological saline and filtered. Mixing the filtrate to prepare a hairy mildew spore suspension, and putting the hairy mildew spore suspension in a refrigerator at 4°C (2);

Select a yellow bean with a large granule and a uniform size, weigh and wash thoroughly, mix with water at a volume ratio of 1:3, soak at 30°C for 3 hours, drain the water, and then use a high-temperature autoclave, at 121°C. Steaming for 30 minutes and then cooling (3); And

According to the mass ratio of 1:3, a molecular blocking amount of 300 kDa was inoculated with a polysaccharide blocking solution and a fungal spore suspension, incubated for 48 hours at a temperature of 25 ℃, and then subjected to post-ripening treatment for 48 hours, It includes a mixed inoculation step (4) to obtain a mushroom polysaccharide 300 kDa blocking liquid Douchi fermentation product.

In step (1) of this embodiment, the leaching temperature in the water bath is 70°C to 85°C.

In step (1) of this embodiment, distilled water is added to the centrifugation supernatant, and the black ear mushroom polysaccharide in the supernatant is diluted to 4 g/L.

In step (2) of the present embodiment, per 200 g of boiled and cooked yellow bean, a total of 2 ml of a wood ear mushroom polysaccharide blocking solution and a hairy fungus spore suspension are inoculated.

In this embodiment, the concentration of spores in the hairy fungus spore suspension is 1.0 Х10 ⁷ ~ 1.0 Х10 ⁸ /ml.

In specific embodiment 6, the method for producing the composition for treating hair loss of this specific embodiment,

Wash the black ear mushrooms thoroughly, remove impurities, and screen with 40 mesh after crushing to prepare black ear mushroom powder, and add black ear mushroom powder in distilled water according to the 140:1 mass ratio of black ear mushroom powder and distilled water. , Ultrasound is performed for 15 minutes under the condition that the ultrasonic power is 500 W, and after leaching in a water bath for 3 hours, the extract is centrifuged at 4500 r/min for 10 minutes to remove impurities, and then the extract is used as black ear mushrooms. Prepared as a material solution with a polysaccharide concentration of 4 g/L and a pH value of 5.6, and then, ultrafiltration is performed under conditions of a temperature of 40 °C and a pressure of 0.17 bar, and a blocking solution of a wood ear mushroom polysaccharide with a molecular blocking amount of 300 kDa. Membrane separation step (1) of the polysaccharide of wood ear mushrooms to obtain

Steps to obtain a fermentation substrate by selecting and weighing and washing yellow bean of uniform size with rich granules, mixing with water at a volume ratio of 1:3, immersing in water for 10 to 18 hours, and sterilizing at 120 ℃ (2 ); And

The fermentation substrate is inoculated into the complex solution until the concentration of the complex solution in the fermentation substrate becomes 2 ml/L, and the complex solution is mixed with the Bacillus inoculum in a mass ratio of 1:3 with a 300 kDa blocking solution of a wood ear mushroom polysaccharide. , Shaking evenly, and incubating at 4 to 8° C. for 72 to 120 hours, and obtaining a Bacillus balance-neck ear mushroom polysaccharide 300 kDa blocking liquid Douchi fermentation product (3).

In step (1) of this embodiment, the leaching temperature in the water bath is 70°C to 85°C.

In step (1) of this embodiment, distilled water is added to the centrifugation supernatant, and the black ear mushroom polysaccharide in the supernatant is diluted to 4 g/L.

In this embodiment, the spore concentration of the Bacillus inoculum inoculum is 1.0 Х10 ⁷ to 1.0 Х10 ⁸ /ml.

The effect of the present invention is verified through the following experiment.

In Example 1, the method of preparing a composition for treating hair loss,

Wash the black ear mushrooms thoroughly, remove impurities, and screen with 40 mesh after crushing to prepare black ear mushroom powder, and add black ear mushroom powder in distilled water according to the 140:1 mass ratio of black ear mushroom powder and distilled water. , Ultrasound is performed for 15 minutes under the condition that the ultrasonic power is 500 W, and after leaching in a water bath for 3 hours, the extract is centrifuged at 4500 r/min for 10 minutes to remove impurities, and then the extract is used as black ear mushrooms. Prepared as a material solution with a polysaccharide concentration of 4 g/L and a pH value of 5.6, and then, ultrafiltration is performed under conditions of a temperature of 40 °C and a pressure of 0.17 bar, and a blocking solution of a wood ear mushroom polysaccharide with a molecular blocking amount of 300 kDa. Membrane separation step (1) of the polysaccharide of wood ear mushrooms to obtain And

Select a yellow bean with a large granule and a uniform size, weigh and wash thoroughly, mix with water at a volume ratio of 1:3, soak at 30°C for 3 hours, drain the water, and then use a high-temperature autoclave, at 121°C. Steamed at for 30 minutes, cooled, and put in a polysaccharide blocking solution of wood ear mushrooms having a molecular blocking amount of 300 kDa, and incubated for 48 hours at a temperature of 25° C. to obtain a fermentation product before tofu; And a step (2) to obtain a fermented product of 300 kDa blocking liquid of Hwangdu-neck ear mushroom polysaccharide by post-ripening treatment for the next 48.

In step (1) of this example, the leaching temperature in the water bath is 70°C to 85°C.

In step (1) of this example, distilled water is added to the centrifugation supernatant, and the black ear mushroom polysaccharide in the supernatant is diluted to 4 g/L.

In Example 2, the method of preparing a composition for treating hair loss,

Wash the black ear mushrooms thoroughly, remove impurities, and screen with 40 mesh after crushing to prepare black ear mushroom powder, and add black ear mushroom powder in distilled water according to the 140:1 mass ratio of black ear mushroom powder and distilled water. , Ultrasound is performed for 15 minutes under the condition that the ultrasonic power is 500 W, and after leaching in a water bath for 3 hours, the extract is centrifuged at 4500 r/min for 10 minutes to remove impurities, and then the extract is used as black ear mushrooms. Prepared with a material solution having a polysaccharide concentration of 4 g/L and a pH value of 5.6, and then performing ultrafiltration under conditions of a temperature of 40° C. and a pressure of 0.17 bar, to select a 300 kDa permeate of a tree ear mushroom polysaccharide, Membrane separation step of polysaccharides of wood ear mushrooms (1); And

Select a yellow bean with a large granule and a uniform size, weigh and wash thoroughly, mix with water at a volume ratio of 1:3, soak at 30°C for 3 hours, drain the water, and then use a high-temperature autoclave, at 121°C. Steamed for 30 minutes at, then cooled, put in the permeate of 300 kDa of wood ear mushroom polysaccharide, and incubated for 48 hours at a temperature of 25° C. to obtain a fermentation product before topping; It includes a step (2) to obtain a fermented product of 300 kDa permeate of Hwangdu-neck ear mushroom polysaccharide by post-ripening treatment for the next 48.

In step (1) of this example, the leaching temperature in the water bath is 70°C to 85°C.

In step (1) of this example, distilled water is added to the centrifugation supernatant, and the black ear mushroom polysaccharide in the supernatant is diluted to 4 g/L.

In Example 3, the method of preparing a composition for treating hair loss,

Wash the black ear mushrooms thoroughly, remove impurities, and screen with 40 mesh after crushing to prepare black ear mushroom powder, and add black ear mushroom powder in distilled water according to the 140:1 mass ratio of black ear mushroom powder and distilled water. , Ultrasound is performed for 15 minutes under the condition that the ultrasonic power is 500 W, and after leaching in a water bath for 3 hours, the extract is centrifuged at 4500 r/min for 10 minutes to remove impurities, and then the extract is used as black ear mushrooms. Prepared as a material solution with a polysaccharide concentration of 4 g/L and a pH value of 5.6, and then, ultrafiltration is performed under conditions of a temperature of 40 °C and a pressure of 0.17 bar, and a blocking solution of a wood ear mushroom polysaccharide with a molecular blocking amount of 300 kDa. Membrane separation step (1) of the polysaccharide of wood ear mushrooms to obtain

Put the hairy fungus species in sterile physiological saline, absorb the bacteria solution with a pipette and inoculate it into the bran medium, incubate for 96 hours at 20°C, and take out the bran inside the Erlenmeyer flask after full growth of mycelium and gray spores, and prepare for use. , 500 mL of sterile physiological saline was added to the cultured mother starter culture bottle and sufficiently shaken, filtered with double gauze, and the filtered residue was washed once again with 500 mL of sterile physiological saline and filtered. Mixing the filtrate to prepare a hairy mildew spore suspension, and putting the hairy mildew spore suspension in a refrigerator at 4°C (2);

Select and weigh and clean yellow bean with a large granule and uniform size, mix with water at a mass ratio of 1:3, soak for 3 hours at 30°C, drain the water, and in a high-temperature autoclave, 121°C Steaming for 30 minutes, and then cooling (3), and

According to the mass ratio of 1:3, a molecular blocking amount of 300 kDa was inoculated with a polysaccharide blocking solution and a fungal spore suspension, incubated for 48 hours at a temperature of 25 ℃, and then subjected to post-ripening treatment for 48 hours, And a mixed inoculation step (4) to obtain a mushroom polysaccharide 300 kDa blocking liquid Douchi fermentation product.

In step (1) of this example, the leaching temperature in the water bath is 70°C to 85°C.

In step (1) of this example, distilled water is added to the centrifugation supernatant, and the black ear mushroom polysaccharide in the supernatant is diluted to 4 g/L.

In step (2) of the present example, per 200 g of boiled and cooked yellow bean, a total of 2 ml of a wood ear mushroom polysaccharide blocking solution and a hairy fungus spore suspension were inoculated.

In this example, the concentration of spores in the hairy fungus spore suspension is 1.0Х10 ⁷ ~ 1.0Х10 ⁸ pcs/ml.

In Example 4, the method of preparing a composition for treating hair loss,

Wash the black ear mushrooms thoroughly, remove impurities, and screen with 40 mesh after crushing to prepare black ear mushroom powder, and add black ear mushroom powder in distilled water according to the 140:1 mass ratio of black ear mushroom powder and distilled water. , Ultrasound is performed for 15 minutes under the condition that the ultrasonic power is 500 W, and after leaching in a water bath for 3 hours, the extract is centrifuged at 4500 r/min for 10 minutes to remove impurities, and then the extract is used as black ear mushrooms. Prepared with a material solution having a polysaccharide concentration of 4 g/L and a pH value of 5.6, and then performing ultrafiltration under conditions of a temperature of 40° C. and a pressure of 0.17 bar, to select a 300 kDa permeate of a tree ear mushroom polysaccharide, Membrane separation step of polysaccharides of wood ear mushrooms (1);

Put the hairy fungus species in sterile physiological saline, absorb the bacteria solution with a pipette and inoculate it into the bran medium, incubate for 96 hours at 20°C, and take out the bran inside the Erlenmeyer flask after full growth of mycelium and gray spores and prepare for use , 500 mL of sterile physiological saline was added to the cultured mother starter culture bottle and sufficiently shaken, filtered with double gauze, and the filtered residue was washed once again with 500 mL of sterile physiological saline and filtered. Mixing the filtrate to prepare a hairy mildew spore suspension, placing the hairy fungus spore suspension in a refrigerator at 4° C., and storing it as a species (2); And

Select a yellow bean with a large granule and a uniform size, weigh and wash thoroughly, mix with water at a volume ratio of 1:3, soak at 30°C for 3 hours, drain the water, and then use a high-temperature autoclave, at 121°C. Steamed for 30 minutes at, then cooled, prepared for use, inoculated with 300 kDa permeate of wood ear mushroom polysaccharide and hairy fungus spores according to a mass ratio of 1:3, and incubated for 48 hours at a temperature of 25° C. Obtaining pre-fermentation products; It includes a step (3) to obtain a fermented product of 300 kDa permeate of hairy fungus-tree mushroom polysaccharide by post-ripening treatment for the next 48 hours.

In step (1) of this example, the leaching temperature in the water bath is 70°C to 85°C.

In step (1) of this example, distilled water is added to the centrifugation supernatant, and the black ear mushroom polysaccharide in the supernatant is diluted to 4 g/L.

In step (2) of the present example, per 200 g of boiled and cooked yellow bean, a total of 2 ml of a wood ear mushroom polysaccharide blocking solution and a hairy fungus spore suspension were inoculated.

The concentration of spores in the hairy fungus spore suspension in this example is 1.0Х10 ⁷ ~ 1.0Х10 ⁸ pcs/ml.

In Example 5, the method of preparing a composition for treating hair loss,

Wash the black ear mushrooms thoroughly, remove impurities, and screen with 40 mesh after crushing to prepare black ear mushroom powder, and add black ear mushroom powder in distilled water according to the 140:1 mass ratio of black ear mushroom powder and distilled water. , Ultrasound is performed for 15 minutes under the condition that the ultrasonic power is 500 W, and after leaching in a water bath for 3 hours, the extract is centrifuged at 4500 r/min for 10 minutes to remove impurities, and then the extract is used as black ear mushrooms. Prepared as a material solution with a polysaccharide concentration of 4 g/L and a pH value of 5.6, and then, ultrafiltration is performed under conditions of a temperature of 40 °C and a pressure of 0.17 bar, and a blocking solution of a wood ear mushroom polysaccharide with a molecular blocking amount of 300 kDa. Membrane separation step (1) of the polysaccharide of wood ear mushrooms to obtain And

Select and weigh and clean yellow beans with large granules and uniform size, mix with water at a volume ratio of 1:3, soak for 3 hours at 30°C, water content 45%-55%, boil and cook, ripe ingredients Was cooled to 30° C. to 40° C., and then inoculated with wood ear mushroom polysaccharide blocking solution and yeast fungus inoculation solution having a molecular blocking amount of 300 kDa at a mass ratio of 1:3, and maintained at room temperature at 28° C., for 22 hours. Koji making) to perform the first soup turnover (turning koji), imperial for 28 hours to perform the second soup turnover, 34 hours to empire and then leave (finished koji), and after leaving the country to wash the soup,12 It includes the step (2) of steaming for a period of time and placing it in a container, heat-retaining fermentation in a constant temperature room of 28° C. for 30 days or more, and then taking out and drying to obtain a fermented product of 300 kDa blocking solution of koji mold-tree ear mushroom polysaccharide.

In step (1) of this example, the leaching temperature in the water bath is 70°C to 85°C.

In step (1) of this example, distilled water is added to the centrifugation supernatant, and the black ear mushroom polysaccharide in the supernatant is diluted to 4 g/L.

In step (2) of this example, per 200 g of boiled and cooked yellow beans, a total of 3 ml of a polysaccharide blocking solution and a yeast fungus inoculum were inoculated, and the concentration of the yeast fungus solution spores was 1.0 Х10 ⁷ ~ 1.0 Х10 ⁸ pcs/ml. .

In step (2) of the present example, "to raise steam" means processing in a closed incubator with a temperature of 30±2°C.

In Example 6, the method of preparing a composition for treating hair loss,

Wash the black ear mushrooms thoroughly, remove impurities, and screen with 40 mesh after crushing to prepare black ear mushroom powder, and add black ear mushroom powder in distilled water according to the 140:1 mass ratio of black ear mushroom powder and distilled water. , Ultrasound is performed for 15 minutes under the condition that the ultrasonic power is 500 W, and after leaching in a water bath for 3 hours, the extract is centrifuged at 4500 r/min for 10 minutes to remove impurities, and then the extract is used as black ear mushrooms. Prepared with a material solution having a polysaccharide concentration of 4 g/L and a pH value of 5.6, and then performing ultrafiltration under conditions of a temperature of 40° C. and a pressure of 0.17 bar, to select a 300 kDa permeate of a tree ear mushroom polysaccharide, Membrane separation step of polysaccharides of wood ear mushrooms (1); And

Select and weigh and clean yellow beans of uniform size with rich granules, mix with water at a volume ratio of 1:3 until the moisture content reaches 50%, soak at 30℃ for 3 hours, boil and cook, and cooked ingredients Was cooled to 35° C., inoculated with 300 kDa permeate of wood ear mushroom polysaccharide and yeast inoculum according to a mass ratio of 1:3, stirred uniformly and placed indoors, maintained at room temperature at 28° C., and imperial for 22 hours. (koji making) to perform the first soup turning (turning koji), empire for 28 hours to perform the second soup reversal, after empire for 34 hours, finished koji, wash the soup after departure, Including the step (2) of steaming for 12 hours and putting it in a container, heat-retaining fermentation in a constant temperature room at 28°C for 30 days or more, and then taking it out and drying to obtain a fermented product of 300 kDa permeate of koji mold-tree mushroom polysaccharide. .

In step (1) of this example, the leaching temperature in the water bath is 70°C to 85°C.

In step (1) of this example, distilled water is added to the centrifugation supernatant, and the black ear mushroom polysaccharide in the supernatant is diluted to 4 g/L.

In step (2) of this example, per 200 g of boiled and cooked yellow beans, a total of 3 ml of a polysaccharide blocking solution and a yeast fungus inoculum were inoculated, and the concentration of the yeast fungus solution spores was 1.0 Х10 ⁷ ~ 1.0 Х10 ⁸ pcs/ml. .

In step (2) of the present example, "to raise steam" means processing in a closed incubator with a temperature of 30±2°C.

In Example 7, the method of preparing a composition for treating hair loss,

Wash the black ear mushrooms thoroughly, remove impurities, and screen with 40 mesh after crushing to prepare black ear mushroom powder, and add black ear mushroom powder in distilled water according to the 140:1 mass ratio of black ear mushroom powder and distilled water. , Ultrasound is performed for 15 minutes under the condition that the ultrasonic power is 500 W, and after leaching in a water bath for 3 hours, the extract is centrifuged at 4500 r/min for 10 minutes to remove impurities, and then the extract is used as black ear mushrooms. Prepared as a material solution with a polysaccharide concentration of 4 g/L and a pH value of 5.6, and then, ultrafiltration is performed under conditions of a temperature of 40 °C and a pressure of 0.17 bar, and a blocking solution of a wood ear mushroom polysaccharide with a molecular blocking amount of 300 kDa. Membrane separation step (1) of the polysaccharide of wood ear mushrooms to obtain

Steps to obtain a fermentation substrate by selecting and weighing and washing yellow bean of uniform size with rich granules, mixing with water at a mass ratio of 1:3, immersing in water for 10 to 18 hours, and sterilizing at 120 ℃ (2 ); And

The fermentation substrate is inoculated into the complex solution until the concentration of the complex solution in the fermentation substrate becomes 2 ml/L, and the complex solution is mixed with the Bacillus inoculum in a mass ratio of 1:3 with a 300 kDa blocking solution of a wood ear mushroom polysaccharide. , Shaking evenly, and incubating at 4 to 8° C. for 72 to 120 hours, and obtaining a Bacillus balance-neck ear mushroom polysaccharide 300 kDa blocking liquid Douchi fermentation product (3).

In step (1) of this example, the leaching temperature in the water bath is 70°C to 85°C.

In step (1) of this example, distilled water is added to the centrifugation supernatant, and the black ear mushroom polysaccharide in the supernatant is diluted to 4 g/L.

In this example, the concentration of spores of the Bacillus inoculum is 1.0 Х10 ⁷ ~ 1.0 Х10 ⁸ pcs/ml.

In Example 8, the method of preparing a composition for treating hair loss,

Wash the black ear mushrooms thoroughly, remove impurities, and screen with 40 mesh after crushing to prepare black ear mushroom powder, and add black ear mushroom powder in distilled water according to the 140:1 mass ratio of black ear mushroom powder and distilled water. , Ultrasound is performed for 15 minutes under the condition that the ultrasonic power is 500 W, and after leaching in a water bath for 3 hours, the extract is centrifuged at 4500 r/min for 10 minutes to remove impurities, and then the extract is used as black ear mushrooms. Prepared with a material solution having a polysaccharide concentration of 4 g/L and a pH value of 5.6, and then performing ultrafiltration under conditions of a temperature of 40° C. and a pressure of 0.17 bar, to select a 300 kDa permeate of a tree ear mushroom polysaccharide, Membrane separation step of polysaccharides of wood ear mushrooms (1);

Steps to obtain a fermentation substrate by selecting and weighing and washing yellow bean of uniform size with rich granules, mixing with water in a ratio of 1:3, immersing in water for 10 to 18 hours, and sterilizing at 120 ℃ (2 ); And

The fermentation substrate is inoculated into the complex solution until the concentration of the complex solution in the fermentation substrate is 2 ml/L, and the complex solution is mixed with the Bacillus inoculum in a mass ratio of 1:3 by the permeate solution of 300 kDa of wood ear mushroom polysaccharide. , Shaking evenly, and incubating at 4 to 8° C. for 72 to 120 hours, thereby obtaining a fermented product of 300 kDa permeate of Bacillus bacillus-tree ear mushroom polysaccharide.

In step (1) of this example, the leaching temperature in the water bath is 70°C to 85°C.

In step (1) of this example, distilled water is added to the centrifugation supernatant, and the black ear mushroom polysaccharide in the supernatant is diluted to 4 g/L.

In this example, the concentration of spores of the Bacillus inoculum is 1.0 Х10 ⁷ ~ 1.0 Х10 ⁸ pcs/ml.

Experiment 1. Inhibition experiment of the oral composition of the present invention against seborrheic mouse hair loss

1. Experimental material

25 mg/1 ml of testosterone propionate injection was purchased from Ningbo No. 2 Hormone Factory (菱Quil-Kul-Kul-Sham), and the batch number is 11025.

Salad oil of 90 ml/bottle was purchased from Arowana Co., Ltd. (Sonjil戴唐掘moo).

The weight of 18-22 g and clean grade adult male B6CBAF1/J mice is Beijing Charles River Experimental Animal Technology Co., Ltd. Soon) 2017-0011), and at 23° C. room temperature and 50 to 60% humidity, ventilate twice a day, and last for 1 hour each time, depending on the natural life mode of the mouse, B6CBAF1/J mice They were allowed to act freely and consume water and food.

2. Experiment method

2.1. Grouping experiments

From 100 male mice, 10 B6CBAF1/J mice were randomly selected and used as an experimental blank control, and the remaining 90 B6CBAF1/J mice were used as experimental mice, separated and bred. That is, it is divided into a hair loss model group and an example 1 group to an example 8 group in which the composition is administered orally.

2.2. Animal model preparation and administration method

The injection modeling method was applied to the hair loss animal model, and the injection site was subcutaneous with the back of the mouse parallel to the spinal column, and in addition to the injection of physiological saline into the blank control group, the remaining 9 groups were injected with a testosterone propio dose of 5 mg/kg·d. Nate is injected, but the injection period is 4 weeks.

At the 4th week of successfully modeling the B6CBAF1/J mouse hair loss animal model, drug treatment was started, and the composition of each example was directly administered intragastrically to all hair loss model groups other than the blank group, but was administered for 6 weeks to treat . Observing the hair growth status of the mouse at the hair loss site of the mouse to calculate the mouse hair loss weight; In connection with pathology observation, the average value was calculated as the ratio of terminal hair/down hair under the microscope when compared with the model group.

3. Data statistical analysis

The data were processed with SPSS 20.0 software, but the difference between groups was analyzed by One Way ANOVA, and p<0.05 represents a significant difference, and it is understood to have statistical significance.

4. Experiment result

(1) hair loss weight measurement

As can be seen from Table 1, compared to the blank control group, a different degree of hair loss occurred in all of the other experimental mice, except that there were significant differences in Examples 3, 5, and 7 (p<0.05). , There was a very significant difference in each of the remaining groups (p<0.01); Compared to the model group, all of the hair loss phenomena of the mice of Example 3, Example 5, and Example 7 were improved (p<0.01), and there was no significant difference in all of the other Example groups, and the improvement trend for hair loss Was not seen.

(2) Measurement of the mouse terminal hair/down hair ratio

As can be seen from Table 1, compared to the blank control group, the terminal hair/fluffy ratio of the model group was significantly reduced (p<0.01); Compared to the model group, Example 3, Example 5, and Example 7 can significantly increase the terminal hair/down hair ratio (p<0.01), and the terminal hair/down hair ratio of other experimental examples was not improved.

(3) Compositions for oral administration of the present invention Examples 3, 5, and 7 have a remarkable improvement effect on seborrheic hair loss, so it will be explained that soybeans can exert a remarkable hair firming effect after fungal fermentation. .

Table 1. Mouse hair loss weight measurement results

Grouping (n=10) Hair loss weight (mg) Blank control 5.03±0.57 Model control 13.78±1.03 ^** Example 1 12.42±1.45 ^** Example 2 13.98±0.74 ^** Example 3 7.56±0.92 ^△△ Example 4 13.07±0.89 ^** Example 5 6.85±0.41 ^△△ Example 6 12.74±1.02 ^** Example 7 5.86±0.48 ^△△ Example 8 12.65±0.97 ^**

** indicates p<0.01 compared to the blank control; ΔΔ indicates that p<0.01 compared to the model group. Table 2. Results of measurement of mouse terminal hair/down hair ratio

Grouping (n=10) ratio Blank control 8.36±0.63 Model control 3.08±0.45 ^** Example 1 3.42±0.71 ^** Example 2 3.28±0.48 ^** Example 3 7.46±0.39 ^△△ Example 4 4.05±0.73 Example 5 7.95±0.81 ^△△ Example 6 3.25±0.30 Example 7 8.23±0.53 ^△△ Example 8 3.31±0.28

** indicates p<0.01 compared to the blank control; △△ indicates that p<0.01 compared to the model group. Experiment 2. Influence of oral administration composition on the amount of hair follicles in mice during hair growth

1. Experimental material

Rosin was purchased from Shenzhen Chitian Chemical Industry Co., Ltd. (深柏市吉田化工有限公司), and pyrapin was purchased from Auwei Adhesive Industrial Wax Products (Zianshi); Mice with a body weight of 18-22 g and clean grade adult magnetic C57BL/6 mice were selected with a pink skin color and resting hair. It was purchased from Beijing Charles River Experimental Animal Technology Co., Ltd. (License No.: SCXK 2017-0149). , Ventilated twice daily, lasted for 1 hour each time, and allowed to freely act and consume water and food according to the natural life mode of the mouse.

2. Experiment method

2.1. Grouping experiments

90 magnetic C57BL/6 mice were randomly divided into 9 groups, but the blank control group and the composition were orally administered in Example 1 to Example 8, separated and reared.

2.2. Animal model preparation and administration method

After mixing rosin and pyrapin 1:1, heating and dissolving, apply evenly on the back of the mouse, but the area of application is about 2 cm Х 3 cm, and after cooling to solidify, remove the back hair (hair It should be confirmed that the growing season is a resting period). The day after hair loss, in addition to direct intragastric administration of distilled water to the blank group, it is administered directly intragastrically to each other group, but is administered once daily at a fixed time, and the volume of direct administration into the stomach is 2 ml/100 g, for 20 consecutive days. Administer. On the 20th day, a piece of skin tissue at the same location on the back of each group of mice was taken and fixed with 10% formalin, and the section was embedded with pyraffin, the thickness of the section was about 4-5 μm, and stained with HE, Statistical processing was performed by taking the average value of the number of hair follicles in the high magnification visual field (Х400).

3. Data statistical analysis

The data were processed with SPSS 20.0 software, but the difference between groups was analyzed by one-way analysis of variance, and p<0.05 indicates a significant difference and is understood to have statistical significance.

4. Experiment result

(1) Effect of hair follicle density

On the 20th day of the experiment, the quantity of hair follicles of mice in each group is shown in Table 3. Compared to the blank control group, Examples 3, 5, and 7 can increase the amount of hair follicles in the mice of the blank group (p<0.01), and the examples of each other group are based on the amount of hair follicles in C57BL/6 mice. Bar not affected (p>0.05); This explains that the soybean fermentation product induces the hair follicles of C57BL/6 mice to enter the growth phase, prolongs the growth phase, delays the entry of the degenerative phase, and promotes the hair follicle growth phase.

Table 3. Mouse hair loss weight measurement results

Grouping (n=10) Hair follicle quantity (pcs/high magnification field of view) Blank control 12.53±1.07 Example 1 12.42±1.25 Example 2 13.05±1.19 Example 3 15.56±0.94 ^** Example 4 11.98±0.74 Example 5 16.23±1.02 ^** Example 6 13.14±1.10 Example 7 16.06±0.85 ^** Example 8 12.84±0.94

* Indicates p<0.05 compared to the blank control group, ** indicates P<0.01 compared to the blank control group.

Claims (10)

As a method for producing a composition for treating hair loss,
Wash the black ear mushrooms thoroughly, remove impurities, and screen with 40 mesh after crushing to prepare black ear mushroom powder, and add black ear mushroom powder in distilled water according to the 140:1 mass ratio of black ear mushroom powder and distilled water. , Ultrasound is performed for 15 minutes under the condition that the ultrasonic power is 500 W, and after leaching in a water bath for 3 hours, the extract is centrifuged at 4500 r/min for 10 minutes to remove impurities, and then the extract is used as black ear mushrooms. Prepared as a material solution with a polysaccharide concentration of 4 g/L and a pH value of 5.6, and then, ultrafiltration is performed under conditions of a temperature of 40 °C and a pressure of 0.17 bar, and a blocking solution of a wood ear mushroom polysaccharide with a molecular blocking amount of 300 kDa. Membrane separation step (1) of the polysaccharide of wood ear mushrooms to obtain And
Select and weigh and clean yellow beans with large granules and uniform size, mix with water at a volume ratio of 1:3, soak for 3 hours at 30°C, boil and cool cooked ingredients to 30°C to 40°C Then, inoculate the polysaccharide blocking solution of wood ear mushrooms with a molecular blocking amount of 300 kDa and the yeast fungus inoculum at a mass ratio of 1:3, maintain room temperature at 28 ℃, and perform koji making for 20 to 24 hours. Perform turning koji, imperial for 28 hours to perform secondary soup overturning, finished koji after imperial for 34 hours, wash the soup after departure, and steam for 12 hours Into, heat-retaining fermentation in a constant temperature room at 28° C. for at least 30 days, and then taken out and dried to obtain a 300 kDa blocking liquid Douchi fermented product, characterized in that it comprises (2) A method of manufacturing a composition for treating hair loss.
The method of claim 1,
In step (2), the method for producing a composition for treating hair loss, characterized in that the moisture content is 50%.
The method of claim 1,
In step (2), a method for producing a composition for treating hair loss, characterized in that the ripe material is cooled to 35°C.
The method of claim 1,
In step (2), the method for producing a composition for treating hair loss, characterized in that performing the first soup overturn by imperial for 22 hours.
The method of claim 1,
In step (2), per 200 g of boiled and cooked yellow beans, a total of 3 ml of a polysaccharide blocking solution and a yeast fungus inoculum are inoculated, and the concentration of the yeast fungus solution spores is 1.0 Х10 ⁷ ~ 1.0 Х10 ⁸ pcs/ml. A method of manufacturing a composition for treating hair loss.
As a method for producing a composition for treating hair loss,
Wash the black ear mushrooms thoroughly, remove impurities, and screen with 40 mesh after crushing to prepare black ear mushroom powder, and add black ear mushroom powder in distilled water according to the 140:1 mass ratio of black ear mushroom powder and distilled water. , Ultrasound is performed for 15 minutes under the condition that the ultrasonic power is 500 W, and after leaching in a water bath for 3 hours, the extract is centrifuged at 4500 r/min for 10 minutes to remove impurities, and then the extract is used as black ear mushrooms. Prepared as a material solution with a polysaccharide concentration of 4 g/L and a pH value of 5.6, and then, ultrafiltration is performed under conditions of a temperature of 40 °C and a pressure of 0.17 bar, and a blocking solution of a wood ear mushroom polysaccharide with a molecular blocking amount of 300 kDa. Membrane separation step (1) of the polysaccharide of wood ear mushrooms to obtain
Put the hairy fungus species in sterile physiological saline, absorb the bacteria solution with a pipette and inoculate it into the bran medium, incubate at 20°C for 96 hours, and add 500 mL of sterile physiological saline to the cultured mother starter culture bottle. Put in, shake sufficiently, filter with double gauze, wash the filtered residue again with 500 mL of sterile physiological saline and filter, mix the two filtrates to prepare a hairy mildew spore suspension, and prepare the hairy fungus spore suspension 4 Putting it in the refrigerator at ℃ (2);
Select a yellow bean with a large granule and a uniform size, weigh and wash thoroughly, mix with water at a volume ratio of 1:3, soak at 30°C for 3 hours, drain the water, and then use a high-temperature autoclave, at 121°C. Steaming for 30 minutes and then cooling (3); And
According to the mass ratio of 1:3, a molecular blocking amount of 300 kDa was inoculated with a polysaccharide blocking solution and a fungal spore suspension, incubated for 48 hours at a temperature of 25 ℃, and then subjected to post-ripening treatment for 48 hours, A method for producing a composition for treating hair loss, comprising a mixed inoculation step (4) to obtain a mushroom polysaccharide 30000 Da duchi fermented product.
The method of claim 6,
In step (2), a method for producing a composition for treating hair loss, characterized in that inoculating a total of 2 ml of a polysaccharide blocking solution and a fungus spore suspension per 200 g of boiled and cooked yellow beans.
As a method for producing a composition for treating hair loss,
Wash the black ear mushrooms thoroughly, remove impurities, and screen with 40 mesh after crushing to prepare black ear mushroom powder, and add black ear mushroom powder in distilled water according to the 140:1 mass ratio of black ear mushroom powder and distilled water. , Ultrasound is performed for 15 minutes under the condition that the ultrasonic power is 500 W, and after leaching in a water bath for 3 hours, the extract is centrifuged at 4500 r/min for 10 minutes to remove impurities, and then the extract is used as black ear mushrooms. Prepared with a material solution with a polysaccharide concentration of 4 g/L and a pH value of 5.6, and then, ultrafiltration is performed under conditions of a temperature of 40°C and a pressure of 0.17 bar, and a blocking solution of throat mushroom polysaccharide with a molecular blocking amount of 30000 Da Membrane separation step (1) of the polysaccharide of wood ear mushrooms to obtain
Steps to obtain a fermentation substrate by selecting and weighing and washing yellow bean of uniform size with rich granules, mixing with water at a volume ratio of 1:3, immersing in water for 10 to 18 hours, and sterilizing at 120 ℃ (2 ); And
The fermentation substrate is inoculated into the complex solution until the concentration of the complex solution in the fermentation substrate becomes 2 ml/L, and the complex solution is mixed with the Bacillus inoculum in a mass ratio of 1:3 with a 300 kDa blocking solution of a wood ear mushroom polysaccharide. , Shaking uniformly, and incubating at 4 to 8° C. for 72 to 120 hours, obtaining a fermented product of Bacillus bacillus-tree ear mushroom polysaccharide 300 kDa blocking solution Douchi fermentation (3). .
The method of claim 8,
A method for producing a composition for treating hair loss, characterized in that the spore concentration of the Bacillus inoculum is 1.0Х10 ⁷ ~ 1.0Х10 ⁸ pcs/ml.
The method of claim 1, 6 or 8,
In step (1), the leaching temperature in the water bath is 70 ℃ ~ 85 ℃ method for producing a composition for treating hair loss, characterized in that.