KR101704645B1 - Fermented Food for Preventing Hair Loss or Promoting Hair Growth - Google Patents

Abstract

The present invention relates to a fermented enzyme food having hair loss prevention and hair growth promoting effect and a method for producing the fermented enzyme food. More specifically, the present invention relates to: 1) 2) mixing Baekhwa-sao and gangwi into the above-mentioned matured mixture, inoculating and fermenting Hwangguk and lactic acid bacteria to prepare a mixed fermented product; 3) drying the mixed fermentation product to produce a dried fermentation product; And 4) pulverizing the dried fermented product and processing the product into any one form selected from the group consisting of granules, powders, rings, capsules, and tablets, to prepare a fermented enzyme food having hair loss prevention or hair growth promoting activity ≪ / RTI > The fermentation enzyme food having the hair loss prevention and hair growth promoting effect produced by the production method of the present invention enhances the fermentation efficiency through the combined fermentation of Hwangguk gum and the lactic acid bacteria so that the physiological active material and fiber And the absorption of the body is improved. In addition, it enhances the absorption of useful substances of Bacillus thuringiensis, Angelica gigas, and black beans, which are known to have wool efficacy, as well as excellent digestive enzyme activity and antioxidative activity, , And thus can be usefully used as a health supplement for alopecia sufferers who are stressed by early hair loss.

Description

{Fermented Food for Preventing Hair Loss or Promoting Hair Growth}

The present invention relates to a method for producing a fermented enzyme food having a hair loss promoting effect and hair loss prevention fermented by applying a combined fermentation technology of Aspergillus oryzae and lactic acid bacteria to a mixture of Bacardiaceae, Angelica gigas, And a fermented enzyme food prepared by the above production method.

Hair removal from hair is called hair loss. Factors affecting hair loss include environmental factors such as exposure to climate, light or heat, disease, birth, hormone secretion and change, drug intake, nutritional status There are internal factors. A major hormone involved in the hair loss mechanism is 5-alpha reductase (see Patent Document 1). 5-Alpha Reducing Enzyme is an enzyme that increases sebaceous secretion by converting testosterone, which is a kind of male hormone (androgen) into DHT (dihydrotestosterone), and inhibits 5-alpha reductase in the hair loss prevention and hair growth promoting products A number of products are being sold. In addition to enzymatic action, hair loss can also be caused by malnutrition, scalp drying, and stress (see Non-Patent Document 1). In the case of hair loss due to such a cause, sufficient nutrition, scalp management and prevention of hair loss by ingestion or administration of antioxidants And promote hair growth.

On the other hand, Baekhasuo is a vine plant of Pusan, and it is distributed nationwide including Jeju Island. It is also called a big mock ( Cynanchum wilfordii ), silver mockery, The root is mainly used as a medicinal material, and its efficacy is to make the body healthier and more vigorous, and to treat hemophilia (see Patent Document 2).

Angelica gigas is a medicinal plant that has been used as a traditional prescription for a long time in the Far East region. In addition, Angelicae is used for anti-inflammatory, diuretic, germane and fever, rheumatism, bacterial and fungal infectious diseases, and diseases of the urinary system. Ferulic acid, a component of Angelica gigas Nakai, has angiogenic activity (see Non-Patent Document 2).

Black beans are high in protein content and contain various lectins and anthocyanins and exhibit various pharmacological actions such as antioxidation, anticancer, anti-obesity, and immunomodulating action (see Patent Document 3). In addition, black soybean has the effect of improving blood circulation through inhibition of thrombogenesis. It contains vitamin E, vitamin B ₁ , vitamin B ₂ , niacin, unsaturated fatty acid and anthocyanin, which helps vasodilation and blood circulation in peripheral blood vessels . Black beans contain antioxidants, four times that of common beans, and cysteine, an intermediate of keratin synthesis mechanisms. They are rich in genistein, a kind of isoflavone, a plant estrogen.

Rice is a name called '10-minute rice dumpling' made with 9 times of rice brown rice. It refers to the rice skin layer containing 95% of rice nutrients, ie, rice removed from rice gruel and rice gruel during the process. Brown rice is also called "one minute glass" and refers to rice husk that has been stripped from the outer rice husk. Therefore, it can be said that it is a complete food rich in carbohydrate, fat, protein, vitamins, calcium and so on, and the nutrient balance through the complete food, brown rice, and various physiological activity effects Can be expected. In particular, dietary fiber among the components of brown rice has been known to exhibit constipation, prevention of postprandial hyperglycemia, prevention of colorectal cancer, and tocotrienol exhibit antioxidative, UV-protecting and cholesterol-increasing effects (see Non-Patent Document 3). Gamma-orizanol has active oxygen elimination, sebum secretion and blood circulation promotion effect, and phytic acid has ability of fat burning action, anti-cancer, strengthening of kidney and liver function, prevention of anemia, prevention of hypertension and inhibition of melanin pigment formation, Blood circulation improvement, neutral fat suppression, obesity suppression, skin improvement, blood pressure improvement, deodorization, nerve sedation, prevention and improvement of dementia. The inositol contained in the brown rice is fatty liver, arteriosclerosis, prevention, Peru rigs acid radicals removal, melanin generation inhibition, oktakosanol cholesterol reduction, fatigue caused by glycogen increases, vitamin B ₂ promotes cell division and vitamin B ₆ are epidermal cells It is known that vitamin E promotes the metabolism of raw materials, vitamin E inhibits active oxygen, potassium prevents hypertension, calcium prevents osteoporosis, zinc prevents arteriosclerosis, magnesium prevents heart disease and iron prevents anemia.

As a result of efforts to produce a fermented product having the effect of preventing hair loss and promoting hair growth by utilizing brown rice and rice bran, the present inventors have applied a combined fermentation technology of Hwang guk gyun and lactic acid bacteria to a mixture of white pearl, angelica, black bean, brown rice and rice bran, As a result, digestion is promoted by digestive enzymes, and absorption of nutrients and functional ingredients of brown rice and rice ghast having a low absorption rate in the body is increased, so that the body utilization rate is increased, the antioxidant effect is excellent, and the fermentation And thus the present invention has been completed.

Korean Patent Publication No. 10-2008-0077762 Korean Patent Publication No. 10-2002-0029412 Korean Patent Publication No. 10-2005-0050962

 Journal of the Korean Society for Design Culture, Vol. 18, No. 2, p. 89-100, 2012  Korean Journal of Oriental Medical Care, Vol. 10, No. 2, p. 189-197, 2002  Korean Journal of Life Science, Vol. 17, No. 8, p. 1141-1146, 2007

The object of the present invention is to provide a method for preventing and treating hair loss by promoting digestive enzyme activity, antioxidant substances and antioxidant effects by applying a combination fermented Hwang gukyun and lactic acid bacterium to a mixture of Baekhasuo, Angelica, Black bean, And a fermented enzyme food having the same.

In order to achieve the above object,

1) mashing black bean, brown rice and rice bran mixture to make a mature mixture;

2) mixing Baekhwa-sao and gangwi into the above-mentioned matured mixture, inoculating and fermenting Hwangguk and lactic acid bacteria to prepare a mixed fermented product;

3) drying the mixed fermentation product to produce a dried fermentation product; And

4) A method for producing a fermented enzyme food having hair loss-preventing or hair growth promoting activity, comprising the step of grinding the dried fermented product and then processing the product into any one form selected from the group consisting of granules, powders, rings, capsules and tablets .

According to one embodiment of the present invention, the mixture of the bagasse, Angelica gigas, black beans, brown rice, and rice bran can be in the form of a powder obtained by pulverizing bagasse, angelica, black bean, brown rice and rice bran, have. The black beans, brown rice and rice bran can be used after harvest regardless of the period. The bagasse and Angelica gigas are preferably pulverized powder forms, but are not limited thereto. The bagasse and Angelica gigas are not limited to powdery ones or powdery ones obtained by drying the extracts. The "extract" includes without limitation any substance extracted from the raw material in any way, and is used to include without limitation all of the extracted extract, the concentrate obtained therefrom, the dried product and the powder of the concentrate.

According to another embodiment of the present invention, the bagasse may be contained in an amount of 1 to 30 parts by weight based on 100 parts by weight of the mixture of the bagasse, Angelica keiskei, black beans, brown rice, and rice bran, And 1 to 20 parts by weight of the bagasse, and 1 to 15 parts by weight of the bagasse, preferably 1 to 10 parts by weight of the bagasse. It is not. If the amount of the bagasse is outside the range of 1 to 30 parts by weight, that is, if the bagasse is included in an amount exceeding the upper limit value, since the bagasse is included more than necessary, the cost of raw material is increased and the production cost is increased.

The Angelica japonica may be contained in an amount of 1 to 20 parts by weight, and the Angelica japonica may be contained in an amount of 1 to 15 parts by weight, based on 100 parts by weight of the mixture of the white algae, Angelica japonica, black beans, brown rice, May be contained in an amount of 1 to 10 parts by weight, and the Angelica japonica may be included in an amount of 1 to 5 parts by weight, preferably 1 to 3 parts by weight, but is not limited thereto. When the Angelica gigas is contained in an amount of 1 to 20 parts by weight or more, that is, when it is contained in excess of the upper limit value, the Angelica gigas are included more than necessary, so that the raw material cost is increased and the production cost is increased.

The black beans may be contained in an amount of 1 to 50 parts by weight, and the black beans may be included in an amount of 1 to 40 parts by weight, based on 100 parts by weight of a mixture of the white bagasse, Angelica gigas, The black beans may be included in an amount of 1 to 30 parts by weight, and the black beans may be included in an amount of 1 to 20 parts by weight, preferably 1 to 15 parts by weight, but are not limited thereto. When the black beans are contained in an amount of 1 to 50 parts by weight or more, that is, the content is less than the lower limit value, the phytoestrogen content and the 5-alpha reductase inhibitory effect in the fermented product are lowered, thereby deteriorating hair loss prevention and hair growth promoting effect.

The brown rice may be contained in an amount of 1 to 70 parts by weight, and the brown rice may be included in 1 to 60 parts by weight with respect to 100 parts by weight of a mixture of the mixture of white saury, Angelica gigas, black beans, brown rice and rice bran. May be contained in an amount of 1 to 50 parts by weight, and the brown rice may be included in 1 to 45 parts by weight, preferably 1 to 35 parts by weight, but is not limited thereto. When the brown rice is contained below the lower limit, the physiological activity of brown rice can not be obtained because functional components such as tocotrienol, gamma-orizanol, and phytic acid are not sufficiently contained.

The raw rice bran may be contained in an amount of 1 to 70 parts by weight based on 100 parts by weight of a mixture of the white birch, Angelica gigas, black beans, brown rice, and rice bran, and the raw rice bran may be contained in an amount of 1 to 65 parts by weight, May be contained in an amount of 1 to 55 parts by weight, and the raw steel may be included in 1 to 45 parts by weight, preferably 1 to 36 parts by weight, but is not limited thereto. When the rice bran is contained below the lower limit value, the polysaccharide component and phenolic acid contained in rice bran are not sufficiently contained in the mixture, so that the physiological activity effects such as immunological activity, anti-cancer, antiallergic and antioxidation can not be sufficiently obtained.

According to one embodiment of the present invention, 1 to 30 parts by weight of the bagasse, 1 to 20 parts by weight of Angelica keiskei koidz., 1 to 20 parts by weight of Angelica keiskei koidz. Are contained in 100 parts by weight of a mixture of the bagasse, Preferably, the black beans are contained in an amount of 1 to 50 parts by weight, the brown rice is contained in an amount of 1 to 70 parts by weight, and the rice bran is contained in an amount of 1 to 70 parts by weight.

According to another embodiment of the present invention, in the step 1), the mixture of black soybean, brown rice and rice bran is preferably mixed at a temperature of 100 to 120 ° C for 0.5 to 3 hours, When used, the steaming step may be omitted. By mixing the black beans, brown rice and rice bran mixture, it prevents bacterial growth through germicidal effect and luxurious action, and creates an environment in which Hwang Kyun-kun and Lactobacillus can grow better.

In the fermentation of step 2), as for Hwang guk gyun and lactic acid bacteria used in fermentation, the live bacteria cultured in a natural medium may be inoculated intact, or powdered strains such as Hwang guk gyong spore powder and freeze-dried lactic acid bacteria powder may be used. The Hwang guk gyun and the lactic acid bacterium can be inoculated at 0.01 to 2.0 parts by weight, preferably 0.05 to 1.5 parts by weight, based on 100 parts by weight of the whole mixture obtained by mixing the bagasse and Angelica with the matured mixture of the step 1) And more preferably 0.1 to 1.0 part by weight, although not limited thereto. When the above Hwang guk gyun and lactic acid bacteria are contained in an amount less than 0.01 part by weight based on 100 parts by weight of the whole mixture of white sachet and Angelica gigas in the matured mixture of step 1), 28 to 35 ° C, preferably 30 to 35 ° C , More preferably at 30 to 32 ° C for 48 to 72 hours, the digestive enzyme activity and antioxidative effect of the final fermentation product can not be sufficiently obtained. If the fermentation product contains more than 2.0 parts by weight of the final fermented product, The amount of the mixture of white sachet, angelica, black bean, brown rice, and rice bran added as a substrate is relatively small, so that the microorganisms enter the stationary phase more quickly than when they are inoculated in the range of 0.01 to 2.0 parts by weight The amount of respiration that occurs at the beginning of fermentation is relatively large, so that the temperature rises due to the respiratory heat, Can be reduced.

Further comprising the step of adjusting the water content so that the moisture content of the whole mixture obtained by mixing the bagasse and Angelica with the matured mixture before fermentation after inoculation of the strain is 55 to 60 parts by weight per 100 parts by weight of the whole mixture, It is desirable to increase the efficiency. When using liquid cultures of Hwanggyu gum and lactic acid bacteria, the total moisture content should be controlled considering the moisture content of the culture. The present fermentation is a solid culture which does not include a stirring process carried out for the purpose of lowering the temperature of the fermentation product increased by the fermentation heat during the oxygen supply and fermentation. The fermentation is continued in the stagnant state until the end of fermentation, And lactic acid bacteria in anaerobic lactic acid bacteria, for example, Lactobacillus and / or Bifidobacterium are simultaneously fermented to induce fermentation efficiency.

When the lactobacillus lactic acid bacterium used in the production of the fermented enzyme food of the present invention is a microorganism belonging to the genus Lactobacillus, the type of the lactobacillus is not particularly limited, but Lactobacillus casei , Lactobacillus acidophilus Lactobacillus acidophilus , Lactobacillus helveticus , Lactobacillus salivarius , Lactobacillus gasseri , Lactobacillus fermentum , Lactobacillus reuteri , Lactobacillus spp ., Lactobacillus spp . Lactobacillus crispatus , Lactobacillus delbruechii subsp . Lactobacillus delbrueckii subsp. Bulgaricus, Lactobacillus delbruecki subsp . Lactobacillus delbrueckii subsp. Delbrueckii , Lactobacillus johnsonii , and the like. Among these bacteria belonging to the genus Lactobacillus, it is particularly preferable to use Lactobacillus casei or Lactobacillus acidophilus in order to enhance digestive enzyme activity and antioxidative effect of the fermented enzyme food of the present invention.

If the bacterium belonging to the genus Bifidobacterium used in the production of the fermented food of the present invention is a microorganism belonging to the genus Bifidobacterium, the species is not particularly limited, but Bifidobacterium breve , Bifidobacterium ronggum (Bifidobacterium longum), Bifidobacterium Infante Tees (Bifidobacterium infantis), Bifidobacterium Adolfo Les sentiseu (Bifidobacterium adolescentis), Bifidobacterium Bifidobacterium Doom (Bifidobacterium bifidum), Bifidobacterium Carthage press ratum (Bifidobacterium catenulatum ), Bifidobacterium pseudocatenulatum , Bifidobacterium angulatum , Bifidobacterium gallicum , Bifidobacterium lactis , Bifidobacterium pseudocatenulatum , Bifidobacterium pseudocatenulatum , Bifidobacterium animalis , and the like. Of these bacteria belonging to the genus Bifidobacterium, it is particularly preferable to use Bifidobacterium bifidum from the viewpoint of enhancing the digestive enzyme activity and antioxidative effect of the fermented enzyme food of the present invention.

The bacterium belonging to the genus Lactobacillus or Bifidobacterium can be easily sold by the Korean Institute of Biotechnology and the Korean Microorganism Conservation Center.

According to one embodiment of the present invention, it is preferable that the lactic acid bacteria are simultaneously fermented with mixed lactic acid bacteria mixed with the lactic acid bacteria of the genus Lactobacillus and Bifidobacterium to induce the fermentation efficiency to be increased. The mixed lactic acid bacteria may be Bifidobacterium bifidum , Bifidobacterium longum and Lactobacillus acidophilus, but are not limited thereto.

According to another embodiment of the present invention, a mixture of Bacillus subtilis, Angelica keiskei, black beans, brown rice and rice bran inoculated with Hwasook Kyun and lactic acid bacteria is heated at 26-40 캜, preferably 28-35 캜, more preferably 30-32 캜 For 36 to 72 hours, preferably 40 to 60 hours, more preferably 48 to 50 hours. Keeping the fermentation temperature at a temperature of 26 to 40 占 폚 is likely to inhibit the growth of lactic acid bacteria or to generate a black streak if the fermentation temperature is 40 占 폚 or higher. If the fermentation temperature is lower than 26 占 폚, the fermentation efficiency is lowered. In addition, by fermentation for 36 to 72 hours, it is possible not only to increase the growth of anaerobic lactic acid bacteria and aerobic bacteria, but also to increase the activity of enzymes (amylase, protease, etc.), polyphenol content and antioxidative activity The absorption rate of nutrients and physiologically active substances in the body can be increased. When the fermentation time is set to 36 to 72 hours, the fermentation can not proceed sufficiently if the fermentation time is less than 36 hours. When the fermentation time exceeds 72 hours, the fermentation time becomes longer and the content of enzymes, antioxidants and other physiologically active substances However, since the raw material of the fermentation, that is, the microorganism, is not endlessly supplied with the substrate used for growth, reproduction, and metabolism, the microorganism can not utilize the substrate as the fermentation time elapses. The fermentation efficiency is lowered, and at the same time, the moisture content in the fermentation chamber is lowered due to the reduction of microbial respiration or the like, and the moisture of the fermentation product is evaporated. It is because.

According to one embodiment of the present invention, the dried fermented product may be pulverized and then further mixed with water to be processed. The fermented product may be pulverized and then processed into a formulation such as granules, powders, pills, capsules or tablets to facilitate ingestion thereof. In addition, although the formulated food may be ingested as it is, it is preferable to undergo a drying process in order to maintain the product shelf life and continuous quality, and it is more preferable to dry the product at a temperature of 40 to 50 ° C. for 5 to 10 hours, It is not.

In addition, the present invention provides a fermented enzyme food prepared by the above production method.

According to one embodiment of the present invention, the fermented enzyme food of the present invention produced by the above production method contains digestion by digestive enzymes, antioxidant effect and antioxidant (see FIGS. 2 to 4). As a result of an animal test and a human body test using a mouse, it was found that the fermented enzyme food prepared according to the above-described production method exhibited hair loss prevention and hair growth promoting effects (see FIGS. 5 to 15). This is because the fermentation enzyme food prepared by the above production method is effective for the fermentation of Hwanggukjun and Lactobacillus bacteria by the combined fermentation of physiologically active substances and nutrients of brown rice and rice bran, The absorption rate of the component can be increased, and the bioavailability can be increased by fermentation.

According to another embodiment of the present invention, the dried fermented product can be crushed, and then the additive and moisture can be further mixed and processed. L-cystine, zinc oxide, vitamin B _6, and dry yeast, which are known to be beneficial to hair health, licorice, green tea, mackerel, red ginseng, pine needles, black sesame seeds, kelp, walnuts Black rice, white rice, black cherry, jujube, earwig, caterpillar fungus, omija, gakgyeong, black rice, ginseng, mung bean, And a concentrated concentrate powder may be used. However, the present invention is not limited thereto, and any material capable of improving the efficacy of the wool without hindering the efficacy of the fermented enzyme food of the present invention can be used without limitation.

Therefore, the production method of the present invention as described above not only raises the absorption rate of the brown rice and the rice ginseng nutrients into the body, but also helps digestion, helps the nutrients of high quality to be absorbed well in the body and has antioxidant activity, prevention of hair loss, Can be usefully used for the production of white seaweed, Angelica gigas, black beans, brown rice and rice bran fermented enzyme foods.

According to the manufacturing method of the present invention, the fermented product of a mixture of Bacillus subtilis, Angelica gigas, Black beans, Brown rice, and Rice bran is obtained by fermenting a mixture of Bacillus subtilis, Angelica gigas, And vitamins, and the enzymes contained in the fermented products and the enzymes produced by the microorganisms change the form of Bacillus thuringiensis, Angelica gigas, black beans, brown rice, and rice bran into digestible forms, thereby facilitating their digestion. In addition, the fermented product according to the production method of the present invention exhibits an effect of preventing hair loss and promoting hair growth, and enhancing absorption of nutrients and functional ingredients of brown rice and rice gut having a low absorption rate in the body, It can help improve health and hair health. In addition, the physiologically active substances contained in Baekhasuo, Angelica gigas, black beans, brown rice and rice bran can show the effect of maintaining health.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a simplified process diagram of a method for producing a fermented enzyme food containing Baek Hwa Sau, Angelica keiskei, black beans, brown rice and rice bran.
FIG. 2 is a view showing the enzymatic activity of the fermented enzyme foods of Baekhwa-sa, angelica, black bean, brown rice and rice bran of the present invention.
FIG. 3 is a graph showing the antioxidant content of the fermented enzyme foods of white shrimp, ginseng, black beans, brown rice and rice bran of the present invention.
Fig. 4 is a graph showing the antioxidant activity of the fermented enzyme foods containing white seaweed, Angelica keiskei, black beans, brown rice and rice bran of the present invention.
Fig. 5 is a photograph showing the visual change of hair regrowth during the test period in the wool efficacy evaluation test using mice of Bacillus thuringiensis, Angelicae radix, black beans, brown rice and rice bran fermented enzyme foods.
FIG. 6 is a graph showing ratios of animals showing the onset of hair regrowth during the test period in the wool efficacy evaluation test using mice of Bacillus thuringiensis, Angelica keiskei, black beans, brown rice and rice bran fermented enzyme foods.
FIG. 7 is a graph showing ratios of animals that reached the hair regrowth filler period during the test period in the wool efficacy evaluation test using the mice of Bacillus thuringiensis, Angelica keiskei, black beans, brown rice and rice bran fermented enzyme foods.
FIG. 8 is a graph showing changes in the hair re-growth area during the test period in the wool efficacy evaluation test using mice of Bacillus thuringiensis, Angelicae radix, black beans, brown rice and rice bran fermenting enzyme foods.
Fig. 9 is a photograph showing histological changes of follicular tissues in a wool efficacy evaluation test using mice of Bacillus thuringiensis, Angelica gigas, black beans, brown rice and rice bran fermented enzyme foods.
Fig. 10 is a microphotograph of the test start and end dates of the human body test of the white seaweed, angelica, black bean, brown rice and rice bran-containing fermented foods of the present invention.
FIG. 11 is a graph showing changes in the total number of dropped hair sensed by a subject during a questionnaire surveyed on subjects who ingested foods of fermented enzyme foods containing white seaweed, Angelica gigas, black beans, brown rice, and rice bran.
FIG. 12 is a graph showing changes in the number of lost hair during a weather condition of a subject during a questionnaire surveyed on subjects who ingested fermented enzyme foods containing white seaweed, angelica, black beans, brown rice, and rice bran of the present invention.
FIG. 13 is a graph showing a change in the number of hairs lost during shampooing of a subject during a questionnaire surveyed on subjects who ingested fermented enzyme foods containing white seaweed, Angelica gigas, black beans, brown rice, and rice bran of the present invention.
Fig. 14 is a graph showing the change in the thickness of hair sensed by a subject during a questionnaire surveyed on a subject who ingested fermented enzyme foods containing white seaweed, ginseng, black beans, brown rice, and rice bran.
FIG. 15 is a graph showing a change in hair growth rate experienced by a subject during a questionnaire surveyed on subjects who ingested foods of fermented enzyme foods containing white seaweed, Angelica keiskei, black beans, brown rice and rice bran.

Hereinafter, the present invention will be described in detail with reference to Examples, Comparative Examples and Experimental Examples.

However, the following examples, comparative examples and experimental examples are intended to illustrate the present invention more specifically, but the scope of the present invention is not limited by these examples and experimental examples.

Example 1. Preparation of fermented enzyme food having hair loss prevention effect and hair growth promoting effect

Black beans, black beans 80-100 mesh, brown rice 80-90 mesh, rice bran 50-60 mesh powder, followed by bagasse, angelica, black beans, black beans, 15 parts by weight of black soybean powder, 36 parts by weight of rice bran, and 35 parts by weight of brown rice were mixed with 100 parts by weight of a mixture of brown rice and rice bran, and the mixture was stirred at 100 to 120 ° C for 0.5 to 3 hours, Were mixed. Then, 10 parts by weight of bagasse powder and 3 parts by weight of Angelica keiskei Powder were mixed and 0.5 part by weight of A. oryzae and 0.5 part by weight of lactic acid bacteria ( B. bifidum , B. longum , L. acidophilus (Vision Biochem)), and the total moisture content was adjusted to 50% by weight. The thus prepared raw materials were fermented for 60 hours in a fermentation chamber at a temperature of 30 DEG C to produce a fermentation product having hair loss prevention effect and hair growth promoting effect. When about 24 hours after the fermentation, the fermentation product turned white due to the proliferation of the bacteria, and when the fermentation time passed 48 hours, the spores began to appear in green. When the fermentation is continued for 60 to 72 hours, the fermentation product is covered with spore spores and becomes green as a whole. Since then, the growth rate of the microorganism is remarkably lowered, and the fermentation product starts drying and the fermentation efficiency is lowered. After fermentation for 60 hours, the fermentation product was dried in a drier at 40 ° C to 50 ° C for 5 to 10 hours to complete the fermentation. Then, after the dried fermentation product was pulverized, 0.019 part by weight of vitamin B ₆ , 0.1 part by weight of zinc oxide, 3.0 parts by weight of dry yeast, 1.0 part by weight of L-cystine and 1 part by weight of a paired concentrate powder and moisture of the fermented product were mixed, Or formulated in the form of a ring. In order to facilitate storage and increase shelf life, the formulated product is finally dried again in a dryer at 40 to 50 ° C to prepare a fermented enzyme food having a moisture content of 10% or less and a hair- , &Quot; fermented product of Example 1 ") was prepared.

Comparative Example 1. Preparation and extraction of a single raw material fermentation product

In order to compare the activity of the fermented product of the single raw material with the fermented enzyme food of Baek Hashuo, Angelica gigas, and black soybean, Baekhasuo, Angelica gigas, and black soybean were mixed with rice bran and fermented. The mixture of the raw materials of Baekhwa-shuo, Angelica gigas, black beans and rice bran was mixed with 40 parts by weight of white beans, Angelica gigas, black bean powder and 60 parts by weight of rice bran, based on 100 parts by weight of the mixture. After 0.5 part by weight of A. oryzae and 0.5 part by weight of lactic acid bacteria ( B. bifidum , B. longum , L. acidophilus ; Vivo Biolchem) were inoculated into the mixture, the total moisture content Was adjusted to 50% by weight. The raw materials thus prepared were fermented in a fermentation chamber at 30 ° C for 60 hours to prepare a fermented product of cereal enzyme using a single raw material.

The resulting product was dried and pulverized. To the pulverized product (300 g) was added 3 L of 70% fermentation alcohol (Barozoro, Daejeon gold, Korea) and stirred at room temperature for 10 hours. The supernatant and precipitate were separated, And extracted. The ethanol extracts were filtered and concentrated using a rotary vacuum evaporator (N-1100, EYELA, Japan) followed by lyophilization (Dry-20R, Ilshin Biotech, Korea).

Experimental Example 1: Determination of digestive enzyme efficacy of fermented enzyme food

1-1. Measurement of amylase activity

The amylase activity of the fermented enzyme food having the hair loss prevention and hair growth promoting effect prepared in Example 1 was measured. In order to investigate the effect of enzyme activity on the enzymatic activity and the effect of Baekhwa - suo, Angelica gigantosa and black bean on the enzymatic activity, the single raw material and brown rice were mixed with each other Amylase activity was compared between the fermented product after fermentation and dried and pulverized white fermented product, Angelica fermented product, black soybean fermented product and its product fermented with brown rice, rice bran and soybean as the main raw materials (product name: Haiyang). Specifically, 1% starch solution and 1% 2,4-dinitrosalicylic acid (DNS) solution were prepared for the experiment, and the experiment was carried out according to the following method. After 3 g of the fermented enzyme food prepared in Example 1, the fermented product of Angelica gigas Fernandes, the fermented Angelica gigas, the fermented black bean product and the product thereof were weighed, 60 ml of a phosphate buffer solution (pH 6.7) was added and stirred at room temperature for 60 minutes The crude enzyme solution was extracted. The extracted crude enzyme solution was centrifuged at 3,200 rpm for 10 minutes to allow the solid powder to settle, and the supernatant was filtered with a Watman filter paper (No. 1) to obtain a test liquid to be used in the experiment. To 0.5 ml of the test solution, 0.5 ml of a 1% starch solution, which had been boiled for 5 minutes at 37 ° C, was added, followed by reaction at 37 ° C for 10 minutes. Then, 1 ml of 1% 2,4-DNS solution was added, and the enzyme activity was stopped by heating at 100 ° C for 5 minutes, followed by allowing to stand at room temperature for 15 to 20 minutes. 0.5 ml of the test solution in which the enzyme activity was inactivated by heating separately at 100 ° C for 5 minutes was used for the blank test by the same method as above. Absorbances of the blank and test groups were measured at a wavelength of 550 nm using a spectrophotometer. Glucose produced from the starch degraded by the enzymatic reaction was colored with a DNS solution and its content was measured from a glucose standard curve prepared in advance. All measurements were repeated three times to calculate mean value and standard error.

As a result, the fermented product of Example 1 of the present invention and the white algae fermented product, the Angelica fermented product, the black soybean fermented product and the amylase activity of the product were found to be 5.15, 4.97, 4.44, 5.00 and 4.45 mg / min · g, respectively (Fig. 2 (a)). It was confirmed that the activity of amylase was higher than that of single fermentation of Bac Hashuo, Angelica gigas, and black soybeans when mixed with fermented product, compared with the existing products that had been fermented with brown rice, rice bran and soybean as main ingredients , And it was confirmed that there was almost no effect on the degradation of enzyme activity by Bacardiaceae, Angelica gigas, and Black bean from the comparison with its products.

1-2. Measurement of protease activity

The protease activity of the fermented enzyme food having the hair loss prevention and hair growth promoting effect prepared in Example 1 was measured. In order to investigate the difference in enzyme activity and the effect of Bacillus subtilis, Angelicae gigantosa, and black soybean on enzyme activity when Baekhwa - sao, Angelica gigas, and black soybean were added, the white algae fermented product, The protease activity of the black bean fermented product and its product was compared. Specifically, the fermentation enzyme food of Example 1, the white fermented product, the fermented Angelica gigas, the fermented black bean product and the product thereof were extracted by the above-described method to obtain a test liquid. For the experiment, 0.6% casein solution, 0.1 N NaOH, 0.4M TCA, 0.4M sodium carbonate solution, 1N Folin reagent and test solution were prepared. The experiment was carried out according to the following method. 3 ml of casein solution and 1 ml of test solution were mixed and reacted at 30 ° C for 10 minutes. As a control, purified water was used instead of the test solution. 5 ml of TCA solution was added thereto to coagulate the remaining protein without decomposition, and the reaction product was filtered with filter paper to remove the undegraded protein which solidified. 1 ml of the filtrate, 2.5 ml of the sodium carbonate solution and 0.5 ml of the pallin reagent were mixed, followed by color development at 30 ° C for 30 minutes, and the absorbance at 660 nm was measured to confirm the content of the degraded protein. Standard curves were prepared using tyrosine, and protease activity was compared and analyzed based on the measured tyrosine content. All measurements were repeated three times to calculate mean value and standard error.

As a result, the protease activity of the fermented product of Example 1 was measured at 188.27 ppm of tyrosine, and the protease activity of the white seaweed fermented product, the black soybean fermented product and the product thereof were 86.28, 258.70 and 203.7 ppm, In the case of water, no protease activity was observed (Fig. 2 (b)). From the above results, it was found that the fermentation efficiency of protease activity was better than that of fermentation of Bacillus subtilis, Angelica gigas and Black beans, respectively.

Experimental Example 2: Antioxidant efficacy of fermented enzyme food

2-1. Determination of polyphenol content

The total polyphenol content of the fermented enzyme food prepared in Example 1 and the total polyphenol content of the fermented product of Baekhwa Sauce, fermented Angelica gigas, black soybean fermented product, and rice fermented product made of brown rice, rice bran and soybean were measured Respectively. Polyphenols are substances that have been reported to reduce the risk of various diseases because of their high antioxidant capacity to protect DNA that is damaged by exposure to reactive oxygen species and cellular constituents and enzymes. There are reports that antioxidative activities such as electron donating ability are increased in proportion to the content of phenolic substances such as polyphenols and flavonoids of natural products. According to these reports, phenolic substances are an indicator for verifying the antioxidative effect of natural products .

The test solution was prepared in the same manner as in the preparation of the test solution for the amylase and protease measurement. To the solution was added 90 ml of purified water and 10 ml of 2N pallin reagent to prepare 0.2 N pallin reagent. 53 g of sodium carbonate was dissolved in purified water, 1 N sodium carbonate solution was prepared so as to be 1 ml. Total polyphenol content was determined by the following method. 450 μl of distilled water and 0.2 ml of 0.2N solution of folin were added to 50 μl of the test solution. The mixture was stirred at 30 ° C for 5 minutes, 2 ml of 1 N sodium carbonate solution was added, and reacted at 30 ° C for 90 minutes. Absorbance was measured at 765 nm . The total polyphenol content was expressed as mg of gallic acid (mg GAE / L) in 1 L of the measurement solution after the calibration curve was prepared using gallic acid as a standard substance. All measurement values were repeated three times and the average value and standard error Respectively.

As a result, the polyphenol content of the fermented product of Example 1 was 1027.60 mg / ml, and the polyphenol contents of the bagasse fermented product, the Angelica fermented product, the black soybean fermented product and the product thereof were 724.65, 645.46, 691.48 and 967.30 mg / , The polyphenol content of the white sachet fermented product, the Angelica fermented product, and the black soybean fermented product was considerably lower than that of the fermented product of Example 1, which was fermented at the same time as the white sake, Angelica gigas, and black soybean (FIG.

2-2. Determination of flavonoid content

Flavonoids, along with polyphenols, are known to be highly antioxidant and effectively remove reactive oxygen species, and are known to have antiviral, anti-inflammatory and anti-cancer effects like polyphenols. Thus, the flavonoid content of the fermented enzyme food prepared in Example 1 of the present invention and the flavonoid content of the fermented product of the fermented product of Baekhwa Sauo fermented product, Angelica fermented product, Black soybean fermented product, Brown rice, The following method was used. The samples to be used for the test were the fermented enzyme foods having the hair loss prevention and hair growth promoting effect prepared in Example 1, 60 g of ethanol 20 times as much as 3 g each of the fermented product of baechasao fermented product, Angelica fermented product, black soybean fermented product, , And then filtered with Wattsman filter paper No. 1. 100 μl of the sample extract was mixed well with 1 ml of diethylene glycol, 100 μl of 1N NaOH was added, and reacted at 37 ° C for 1 hour. Absorbance was measured at 420 nm using a spectrophotometer. The total flavonoid content from the calibration curve obtained in the concentration range of 0 to 0.8 mg / ml using naringin as a reference material is shown in mg of sample g, and the average value and the standard error were calculated by repeating all the measurements three times.

As a result, the total flavonoid content of the fermented product of Example 1, the fermented product of Baekhwa Sauo, the fermented product of Angelica gigantus, the fermented black bean product and the product thereof were 0.1757, 0.1670, 0.1820, 0.1670 and 0.1877 mg / g, respectively (Fig. 3 (b)).

2-3. DPPH test

DPPH analysis is a kind of antioxidant activity measurement method and is mainly used to measure the antioxidant power of phenol and aromatic amine compounds. The following methods were used to measure and compare the antioxidant capacity of the fermented food prepared in Example 1 of the present invention and the antioxidant activity of the white algae fermented product, the Angelica fermented product, the black soybean fermented product and the self fermented food product. The test solution was prepared in the same manner as in the preparation of the test solution for the measurement of amylase and protease, and 0.15 mM (1,1-diphenyl-2-picrylhydrazyl) solution (2.9574 mg of DPPH dissolved in 50 ml of ethanol) Respectively. In the test group (S), 1.9 ml of DPPH solution was added to 100 μl of the test solution, and the reaction solution was wrapped with aluminum foil or the like so as to prevent light from entering into the reaction solution, followed by reaction at 37 ° C for 1 hour. 100 μl of distilled water was added to the blank test group (C), 100 μl of ethanol was added to 100 μl of ethanol, and 100 μl of distilled water was added to 100 μl of the test solution to correct the error that might be caused by the color- After adding 1.9 ml (Co) of ethanol, the reaction was carried out in the same manner as in the test group and the blank test group, and the absorbance values thus obtained were subtracted from the test group and the blank test group, respectively. The free radical scavenging ability (%) was calculated by the following formula, and the experiment was repeated three times and the average and error values were obtained.

Free radical scavenging ability (%) = [{1- (S-So) / (C-Co)}]

As a result, the DPPH activity of the fermented product of Example 1 and the white bait fermented product, the Angelica fermented product, the black soybean fermented product and the product thereof were 93.39, 89.04, 66.86, 64.88 and 91.04%, respectively, The soybean fermented product showed significantly lower DPPH activity than the fermented product of Example 1 (Fig. 4a).

2-4. FRAP test

The ferric reducing antioxidant power (FRAP) test is an antioxidant that measures the reducing power of a test substance using the principle that TPTZ (ferric 2,4,6-tripyridyl-s-triazine) is reduced to Fe ²⁺ Force test method. The following methods were used to measure and compare the antioxidant capacity of the fermented food prepared in Example 1 of the present invention and the antioxidant activity of the white algae fermented product, the Angelica fermented product, the black soybean fermented product and the self fermented food product. The FRAP reaction reagent was prepared by dissolving TPTZ in a 40 mM hydrochloric acid solution at a concentration of 10 mM and adding 300 mM acetate buffer (pH 3.6) corresponding to 10 times the volume of the 10 mM TPTZ solution and 20 mM ferric chloride solution were uniformly mixed and incubated at 37 ° C for 30 minutes.

The test solution for the FRAP test was prepared in the same manner as the test solution for the measurement of amylase and protease. 160 μl of distilled water was added to 40 μl of the test solution, and 1.8 ml of the FRAP reaction reagent was added thereto and the mixture was reacted at 37 ° C. for 10 minutes . Separately, instead of the test solution, 200 μl of distilled water was reacted by the same method. The absorbance of the test group and the test group was measured at 593 nm by a spectrophotometer, and the absorbance of the test group was subtracted from the absorbance of the test group. And eliminate any errors that may occur. The analytical results were expressed as FeSO ₄ · 6H ₂ O as a standard and the absorbance of the sample was expressed as mM FeSO ₄ eq./mgextract. All measurements were repeated three times to obtain a mean value and a standard error .

As a result, the results of the FRAP test of the fermented product of Example 1 and the white fermented product, Angelica gigas Fermented product, black soybean fermented product and its product were 3.22, 1.57, 1.22, 1.07 and 2.18 mM FeSO ₄ eq./mg, respectively, (Fig. 4 (b)). It was confirmed that the reducing power when fermenting Baekhasuo, Angelica gigas and black soybean was significantly higher than that of fermentation of a single raw material.

Experimental Example 3. Evaluation of wool efficacy using a mouse

3-1. Test animal

The wool efficacy of the fermented product was evaluated using a mouse. Six - week - old female C57BL / 6 mice (Korea BioLink; Daeyali, Samsung - si, Chungbuk, Korea) were purchased and subjected to a week - long purifying period. C57BL / 6 mice have a genetic characteristic of spontaneous alopecia with black hair, and melanin cells are restricted to hair follicles, and the melanin synthesis cycle is almost the same as the hair growth cycle, It is possible to determine the growth cycle of hair, and it is the most standardized and broadly studied model as discrimination study and hair research model with the lapse of time over time. It is excellent in reproducibility and stability in experiment application, It is widely used for hair physiology and hair growth promotion research because it has the merit of accumulating basic data of test.

3-2. Animal room environmental conditions

The animal room environment conditions for the experiment are as follows.

end. Breeding cage PSU, 223 × 267 × 145 mm

I. Temperature 21 ± 2 ℃

All. Humidity 50 ± 20%

la. Ventilation system and exhaust before withdrawal, more than 10 times / hr

hemp. Lighting time and contrast 12hr / day / night

                                  (day: 08: 00 ~ 20: 00, night: 20: 00 ~ 08: 00)

bar. Illumination 150 - 300 lux

four. Noise and ammonia concentration Less than 60 dB / 20 ppm

Ah. Feed and and Negative

The rats fed the test animal feed (Agribrands Purina Korea Inc. Analysis Service of Central Laboratory, 627 Jangdang-dong, Pyeongtaek, Kyonggi-do, Korea) were freely taken and fed filtered sterilized purified water in a polycarbonate water bottle (250 ml).

3-3. Test substance

As a test substance, the fermented product of Example 1 and the fermented product of Bacillus thuringiensis, Angelica gigas, and black soybean of Comparative Example 1 were extracted with ethanol and lyophilized, and the wool efficacy with the fermented product of Example 1 Respectively. All samples were dissolved in distilled water and homogenized using an ultrasonic sonicator.

3-4. Configuration of test group

Test groups and daily doses are shown in Table 1 below.

Test group Dose ^a) menstruum Method of administration Diet ^b) Duration of administration ^c) Control group 0 mg / kg Distilled water Oral administration Rats for experimental animals 3 weeks Fermented white sake 300 mg / kg Angelica fermented water Fermented black beans The fermentation product of Example 1 1,000 mg / kg

^{a) The} test substance was orally administered at a dose of 10 ml / kg, and the control group was orally administered only the same amount of distilled water during the test substance administration period.

^b) Rats for experimental animals were purchased from Agribrands Purina Korea Inc. Analysis Service of Central Laboratory (Jangdang-dong, Pyeongtaek, Gyeonggi-do).

^{c) At the time of} receipt, the animals were subjected to a purification process for one week, and the animals were separated and subjected to this test for 3 weeks after epilation.

3-5. Test Methods

3-5-1. Quarantine and purification

After obtaining the animals, veterinary quarantine was carried out and a 7-day purification period was provided. It was used as data to select the healthy animals suitable for the test through the general symptom observation during the purifying period.

3-5-2. waxing

(EN143, Panasonic, Japan) at the end of purification (49 to 50 days of age) the day before the start of the test. After epilating, Hair removal cream (Niclean, Ildong Co., Ltd, Korea) was applied and gauze with water was applied. The length from the back side of the neck to the base of the tail and the width of 1 cm or more Secondary hair removal was performed and only individuals showing a homogeneous pink color on the back skin were selected and used in the experiment.

3-5-3. Group separation

Group separation was carried out the day after epilation, and grouping was performed according to the Z-array method on the basis of body weight only using animals that did not develop abrasions or severe erythema during epilation.

3-5-4. Object identification and breeding management

An individual identification card was attached to the breeding box and a tail marking method was used for individual identification. The environment of the breeding room was kept constant at constant temperature (21 ± 2 ℃), humidity (50 ± 20%) and 12 hours interval (08: 00 ~ 20: 00) And were housed and maintained at the breeding density. For the litter, soft litter for laboratory animals was used. The breeding box was changed once a week. All equipment and litter used in the breeding room were sterilized by high pressure steam sterilization at 121 ° C for 20 minutes.

3-5-5. Administration of the test substance

From the day of group separation (day 0), the test substance was homogenized in distilled water and orally administered at a dose of 10 ml / kg once a day for 21 days. In the control group, only the same amount of distilled water was orally administered once a day for 21 days. The experimental animals in each group were sacrificed for 21 days after fasting for 6 hours and used for histological evaluation analysis.

3-5-6. Clinical symptom observation

At least once a day during the study period, individual symptoms were observed for clinical symptoms.

3-5-7. Visual changes of photography and hair re-growth

(Canon EOS 30D / Canon / Japan) for 9 times from 0, 11, 12, 13, 14, 16, 18, 19, In order to minimize the number of photographs, a photograph was taken under the light of 200 to 300 lux at the designated place (Copy stand CS-30, LPL, Japan). The conditions for the photographing are the same as the distance from the subject to 55 ㎝, and the canon zoom lens (EF 28-105 ㎜ 1: 3.5-4.5 Ⅱ) Center subject focus, auto white balance, no-flash) and recorded as a 'jpg' file with a digital image of 3504 × 2336. The animals used for photography were photographed with lightly anesthetized with dimethyl ether to include five animals in one photograph. Photographs were taken under certain conditions (illumination, height) as stated above, and photographs taken included the markings of individuals and scale bar (in mm).

As a result, the gross changes in hair regrowth over a period of 9 days from 0, 11, 12, 13, 14, 16, 18, 19 and 21 days after 4 weeks of transdermal application of the test substance showed that hair removal After 11 days, after hair growth started at the hairy tail side and neck area, hair growth line was formed on the 15th day and hair regrowth pattern was observed along the spine. (Fig. 5).

3-5-8. Hair growth initiation time and Hair growth completion time Average days calculation

The minimum point at which the hair can be clearly observed at the hair removal site is recorded as individual hair growth regeneration period by visual observation once a day and the minimum point at which the hair regrowth time can be observed at 80% And 'hair fullness period'. However, in the case of the hair filler, considering the error in the area measurement and the occurrence of damaged skin tissue in the hair removal, the hair reapplication time point was defined as 80% or more of the total hair removal area. Based on the obtained results, hair regrowth initiation period was expressed as average days of opening period, and hair - filling period was expressed as the ratio of hair - full - number of animals among all animals on day 21 after hair removal.

The average number of days of hair regrowth initiation in each group and the hair regrowth fill rate of the test end date are shown in Table 2 below.

Test group Hair re-growth Hair development (days) ^a Hair fullness (%) ^b Control group 13.13 ± 2.10 37.5 Fermented white sake 12.13 + - 0.64 75.0 Angelica fermented water 11.88 0.35 75.0 Fermented black beans 11.88 ± 0.83 75.0 The fermentation product of Example 1 11.88 ± 0.64 100.0

^a mean ± standard deviation (n = 8)

^b Rate of mouse reached hair fullness (n = 8)

As a result of the analysis of the results of accumulation of different individuals in hair re-growth initiation period and hair re-growth period, hair re-growth initiation days were observed in all the groups from the 11th day after hair removal, Of the control group, 13.13 ± 2.10 days in the control group, 12.13 ± 0.64 days in the bagasse fermented product group, 11.88 ± 0.35 days in the Angelica fermented product group, 11.88 ± 0.83 days in the black soybean fermented product group, and 11.88 ± 0.64 days in the fermented product group in Example 1 The tendency of initiation was fastest in the order of 'fermented water administration group = Angelica fermented water treatment group = black soybean fermented water treatment group> white marrow fermented water treated group> control group' (Fig. 6).

In the time-dependent analysis results obtained by accumulating other individuals in the hair regrowth filler period, full-fledged individuals appeared in the black soybean fermented product, the Angelica fermented product, and the fermented product administration group of Example 1 after 15 days of hair removal, Except for the control group, over 75% of all test substance-treated animals reached hair regrowth fullness. The proportion of animals that had reached the rehabilitation period of hair after 21 days of hair removal was found to be 37.5% in the control group, 75.0% in the bagasse fermented product group, 75.0% in the Angelica fermented product group, 75.0% in the black soybean fermented product group, (Fig. 7). The results are shown in Fig. 7, in which the fermented water-treated group of Example 1, the Angelica fermented group, the black soybean fermented group,

3-5-9. Change of hair regrowth area

The percentage of hair growth in the entire hair removal area was calculated as a percentage (%) using image analysis software (ImagePartner / SaramSoft / Korea) for each image file obtained from the photo evaluation. The total hair removal area was measured at each calculation to prevent the total hair removal area from varying from each photograph due to the change of the posture of the animal during photography.

Hair regrowth area / hair removal area (%), mini = 0, max = 100%

The ratio of the hair growth area of the total hair removal area (%) of the total hair removal area by using the digital image file obtained by taking a total of 9 photographs of 0, 11, 12, 13, 14, 16, 18, 19, ) Of the re-growth area is shown in Table 3 below.

Hair
Re-growth
area(%) Test group Control group Single raw material fermentation product Example 1
Fermentation product Backseason Angelica Black beans 11th ^a 0.06 0.11 0.07 ± 0.19 0.68 ± 1.92 0.45 + 0.71 0.23 + - 0.43 12th 1.13 ± 1.33 1.70 ± 2.25 2.12 ± 3.45 4.14 ± 5.65 2.46 ± 2.93 13th 4.06 + - 4.50 11.27 ± 15.41 9.95 ± 13.79 10.56 ± 12.19 10.07 ± 11.65 14 days 15.41 ± 20.54 28.89 ± 25.81 25.41 + - 20.98 25.33 + - 22.61 32.28 ± 23.61 16th 30.76 ± 26.45 47.72 ± 25.51 50.92 + - 23.87 42.99 + - 21.80 59.30 ± 17.25 ^* 18th 36.86 ± 25.15 55.74 ± 26.86 59.22 ± 23.71 55.23 + - 21.02 74.24 ± 11.15 ^** 19th 50.34 ± 28.51 70.23 + - 19.79 72.59 ± 17.41 69.81 + - 16.31 83.01 + - 12.09 ^* 21st 57.76 ± 29.27 77.78 +/- 14.57 79.18 ± 15.56 77.90 ± 16.49 87.68 + - 8.62 ^*

^a : mean ± standard deviation (n = 8)

^* : Significant difference from control group ( p <0.05 )

^** : Significant difference from control group ( p <0.01 )

As a result, there was no significant difference in hair regrowth area from 25.41 to 32.28% in all groups except the control group (15.41 ± 20.54%) until day 14 after hair removal, but hair regrowth area of the control group from day 16 after hair removal was 30.76 ± 26.45 %, 50.72 ± 23.87%, 42.99 ± 21.80%, and 59.30 ± 17.25%, respectively, of the group treated with the fermented product of Bacillus subtilis and the fermented product of Example 1 were 47.72 ± 25.51%, 50.92 ± 23.87% The area of regrowth of hair began to increase in the treated group compared to the control group, and the area of regrowth of hair was increased in the fermented group of Example 1 as compared to the control and single raw material fermentation.

The hair regrowth area of the control group was 36.86 ± 25.15% on the 18th day after hair removal, 55.74 ± 26.86% for the white sucrose fermented product group, 59.22 ± 23.71% for the Angelica fermented product group, 55.23 ± 21.02% for the black soybean fermented product group, Of the control group and 74.24 ± 11.15% of the control group, respectively. The hair re-growth area of the test group treated with the fermented product of Example 1 was larger than that of the control and single-component fermented product.

The hair regrowth area of the control group was 50.34 ± 28.51%, 70.23 ± 19.79%, and 72.59 ± 17.41%, respectively, and 69.81 ± 16.31% of the black soybean fermented group, The hair regrowth area of Example 1 was 83.01 ± 12.09%, the hair regrowth area was increased in all the test substance-treated groups compared to the control group, and the hair regrowth area of the fermented group of Example 1 was increased compared to the control and single raw material fermentation .

The hair regrowth area of each group was 57.76 ± 29.27% in the control group, 77.78 ± 14.57% in the bagasse fermented product group, 79.18 ± 15.56% in the Angelica fermented product group, 77.90 ± 16.49% in the black soybean fermented product group, The area of regrowth of hair was 35%, 35%, and 40%, respectively, in the fermented product group of Example 1 and 87.68 ± 8.62% of the control group, the fermented product group, the black soybean fermented product group and the black soybean fermented product group, In the fermented water-administered group, the hair regrowth area was increased by 52%, and the hair regrowth area of the fermented water-treated group of Example 1 was increased as compared with the control and single raw material fermentation (FIG. 8).

3-5-10. Histological Changes of Follicular Tissue

The experimental animals were sacrificed by bleeding and then the first hair removal part was cut off. The cut tissues were fixed in 10% neutral buffered formalin for 1-2 days using filter paper (Watman No.2 185 mm, Watman, Germany). Two strips of 0.5 ㎝ and 2.5 ㎝ in length were obtained for each individual, and two strips were placed in one cassette and subjected to general tissue treatment with alcohol and xylene in stages. Paraffin blocks (including two strips) were prepared for each individual, and a continuous slice of 4 μm thick was prepared. After that, stripped strip sections were added to each slide, and deparaffinized and functionalized, followed by hematoxylin and eosin staining. Histological changes of hair follicles such as size of DP, dermal papilla, number and size of hair follicles and growth of hair shaft were measured using an optical microscope (JP / BX51, Olympus, Japan) Respectively.

It is known that the hair follicles grow to the subcutaneous fat layer by the formation of hair follicles by the interaction of epidermal cells of epidermal layer and dermal papilla cells in dermal layer. The histologic findings of the skin at the end of the study (21 days) showed that the number of hair follicles was increased and the size of hair follicles was also increased in all test substance - treated groups compared to the control group. In addition, the hair follicles observed in the control group's skin tissue were mainly observed in the shape of the hair follicles starting from the circular shape to the elliptical shape as they were folded from the dormant period to the growing period, whereas the hair follicles observed in the skin tissue of the test substance- And the length of the moraine was also prolonged. It was observed that the hair follicles were developed to the dermis and epidermis in the order of the fermented product of Example 1, the Angelica fermented product, the black soybean fermented product, and the white algae fermented product. In the subcutaneous layer, a new hair follicle (Fig. 9).

From the above results, it can be seen that, in the model of C57BL / 6J mouse alopecia, the average number of days of hair growth and hair re-growth during the 3 weeks of oral administration of the fermented product of Example 1, The change of hair regrowth area and the histological evaluation showed that hair regrowth tended to be promoted in all the test substance administration groups from the 16th day after hair removal compared to the control group, And significantly promoted hair regrowth.

3-5-11. Statistical analysis

All experimental results were expressed as mean ± standard deviation. For all data, a Turkey test was conducted to compare variance homogeneity. When the variance was homogeneous, one-way analysis of variance (ANOVA) was performed. When significance was observed, Student's t-test was performed to examine the test group.

Experimental Example  4. Human test  Evaluation of hair growth effect

4-1. Selection of subjects

In order to confirm the efficacy of the fermented product of Example 1 to prevent hair loss and stimulate hair growth on the human body, the test subjects were collected and tested for 8 weeks. Sixty male and female alopecia areata were selected, and the subjects were dropped due to ingestion, questionnaires and inadequate photographs. The results were based on the final 24 test data. The subjects of the 24 subjects who ingested the fermented material of Example 1 for 8 weeks (from May 24, 2014 to July 19, 2014) were 20 to 5, 30 to 4, 40 to 14, and 50 1 male, 15 males and 9 females by gender.

4-2. Changes in hair thickness and hair number

Hair thickness and number of hairs were measured using a Dino-Lite Premier Digital Microscope AM4113T (Taiwan), and the thickness and number of hair after microscopic examination were calculated using DinoCapture 2.0 (Version 1.5.5). The same area was photographed with a magnification of 50 × and an area of 744.66 ㎟ (7.7 ㎜ × 5.81 ㎜) at the start and end of the test, and 11 subjects except for 13 people living in the province were examined (FIG. 10). The sex of the subjects was 6 males and 5 females.

Of the 11 subjects, eight subjects had thicker hair, two subjects with thicker hair thicknesses of 25% or more, four subjects with 50% thickening, and two subjects with more than 100% thickening. Of the 8 patients with thicker hair, 4 were males and 4 were females. Overall, 72.73% had thicker hair thickness (Table 4).

Of the 11 subjects, eight subjects had increased hair counts, two with 10% or more hair growth, four with 10% or more and less than 20% increase, one with a 25% increase and one with a 44% increase , And the sex ratios of the eight men with increased hair count were 4 men and 4 women, respectively, as well as hair thickness (Table 5).

Change in hair thickness (mm) Male subject Female subject number 0 parking 8 parking number 0 parking 8 parking M1 0.050 0.078 F1 0.077 0.091 M2 0.059 0.059 F2 0.061 0.053 M3 0.061 0.076 F3 0.075 0.109 M4 0.050 0.075 F4 0.041 0.093 M5 0.055 0.052 F5 0.066 0.092 M6 0.035 0.076

Change in number of hair Male subject Female subject number 0 parking 8 parking number 0 parking 8 parking M1 38 37 F1 47 56 M2 41 43 F2 36 36 M3 30 36 F3 34 49 M4 38 44 F4 32 40 M5 40 44 F5 42 48 M6 37 36

4-3. Poll

In the 4th and 8th weeks of the test, a questionnaire was conducted on the total number of dropped hairs, the number of hairs lost during the weather, the number of hairs lost during shampooing, the thickness of hair, and the change of hair growth rate.

The subjects' responses to changes in the total number of dropped hairs were answered by 10 (41.7%) less or less at 4th week, and 17 (70.8%) at 8th week were less or less (Fig. 11). In the morning, 12 (50.0%) responded to the number of dropped hair, and 17 (70.8%) answered in the 8th week. The response of the subject to the change of hair number 12 (50.0%) answered in the fourth week and seventeen (70.8%) answered in the 8th week (FIGS. 12 and 13).

The subjects who responded that the thickness of the hair felt by the subject felt a little thicker or thicker was answered positively by the longer duration of intake (12.0% at 4th week) and 16 persons (66.7% at 8th week) (37.5%) in the 4th week, and 8 (33.3%) in the 8th week. The respondents who answered that the growth rate of the hair grows faster (Fig. 15).

Claims (10)

1) mashing black bean, brown rice and rice bran mixture to make a mature mixture;
2) mixing Baekhwa-sao and gangwi into the above-mentioned matured mixture, inoculating and fermenting Hwangguk and lactic acid bacteria to prepare a mixed fermented product;
3) drying the mixed fermentation product to produce a dried fermentation product; And
4) A method for producing a fermented enzyme food having hair loss-preventing or hair growth promoting activity, comprising the step of grinding the dried fermented product and then processing the product into any one form selected from the group consisting of granules, powders, rings, capsules and tablets As a result,
1 to 30 parts by weight of the bagasse, 1 to 20 parts by weight of Angelica gigas Nakai, 1 to 50 parts by weight of black beans, 100 parts by weight of black beans, 1 to 70 parts by weight of the brown rice, and 1 to 70 parts by weight of the rice bran are included. delete The method according to claim 1,
Wherein said Hwangkyongguk and lactic acid bacteria are inoculated at 0.01 to 2.0 parts by weight per 100 parts by weight of the matured mixture of step 1). The method according to claim 1,
Further comprising the step of adjusting the water content so that the water content of the whole mixture obtained by mixing the bagasse and Angelica with the matured mixture of step 1) is 55 to 60 parts by weight per 100 parts by weight of the whole mixture, A method for producing a fermentation enzyme food having a promoting effect. The method according to claim 1,
Wherein the step 1) is carried out at a temperature of 100 to 120 ° C for 0.5 to 3 hours, thereby preventing hair loss and promoting hair growth. The method according to claim 1,
Wherein the fermentation of step 2) is carried out at a temperature of 28 to 35 DEG C for 48 to 72 hours. The method according to claim 1,
Wherein the drying of step 3) is carried out at 40 to 50 ° C for 5 to 10 hours to prevent hair loss and to promote hair growth. The method according to claim 1,
Wherein the lactic acid bacterium is a mixed lactic acid bacterium obtained by mixing lactobacillus of the genus Lactobacillus and Bifidobacterium, and a method of producing the fermented enzyme food having hair loss promoting effect. The method of claim 8,
Wherein the mixed lactic acid bacteria are Bifidobacterium bifidum, Bifidobacterium longum, and Lactobacillus acidophilus. A fermented enzyme food having hair loss prevention activity and hair growth promoting effect produced by the production method according to any one of claims 1 and 3 to 9.

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