KR101781954B1

KR101781954B1 - Method for producing mushroom mixed beverage using Lentinula edodes, Phellinus linteus and medical herb extract and mushroom mixed beverage produced by the same method

Info

Publication number: KR101781954B1
Application number: KR1020150091759A
Authority: KR
Inventors: 서경순; 김경제; 진성우; 최봉석; 박태영; 고영우; 김진경; 김아름찬; 김유선; 허창기; 최유진; 양희선; 진희연
Original assignee: 재단법인 장흥군버섯산업연구원
Priority date: 2015-06-29
Filing date: 2015-06-29
Publication date: 2017-09-26
Also published as: KR20170001972A

Abstract

The present invention relates to a method for preparing a mushroom mixture, comprising: (a) preparing a mushroom mixed extract by mixing water and a conditional extract, (b) preparing water extracts of Angelica keiskei koidz., Acanthopanax senticosus, Acanthopanax senticosus, Acanthopanax senticosus, Lycopersicum blooms, and Malt extracts, respectively, followed by extraction; And (c) mixing the mushroom mixed extract prepared in the step (a) and the Angelica keiskei bark extract prepared in the step (b), Sanswu oil extract, Hwanggi extract, bamboo leaf extract, licorice extract and malt extract, followed by heating And a mushroom mixed beverage produced by the method.

Description

TECHNICAL FIELD The present invention relates to a method for producing a mushroom mixed beverage using an elevation, a condition and herbal extract, and a mushroom mixed drink prepared by the method,

The present invention relates to a mushroom mixture comprising a mushroom mixed extract obtained by mixing a mushroom mixed extract with a mushroom mixture, a mushroom extract, a mushroom extract, a mushroom extract, a bamboo mushroom extract, a licorice extract and a malt extract, And a mushroom mixed drink produced by the method.

As the aging of the population is rapidly progressing in the world and the income level is rising, the anti-aging industry is growing fast and the national social importance is increasing. The anti-aging industry consists of anti-aging products, anti-aging medical devices, anti-aging products, anti-aging products and anti-aging services such as health care services and appearance management services. , Which has grown at an annual average rate of 11%.

According to BCC Research, the global market size of anti-aging industries is expected to grow at a CAGR of 11.1% from USD132.5bn in 2006 to USD274.5bn in 2013. As a result of this rapid growth, the aging population is aging and the purchasing power of the elderly is expanding, and there is a high priced anti-aging product and service market for healthy old age. In addition, the demand for anti-aging industry is increasing in the young age as the economic level is improved and interest in health and appearance is increased.

The emergence of anti-aging industry has the effect of reducing social medical expenses by preventing disease through proactive health care and prevention of aging from the viewpoint of economic and social aspect, and on the other hand, This is important because it can be a growth engine that can drive economic growth based on demand.

A clear and common definition of the anti-aging and anti-aging industries has not yet been established, but from a comprehensive standpoint, the anti-aging industry is the product of prevention and management of aging mechanisms and all products that prevent, manage and cure physical diseases Service. &Lt; / RTI > The detailed areas include anti-aging products, anti-aging products, anti-aging cosmetics, and anti-aging medical device industry. As for the aging industry, The market for aging drugs is expected to reach $ 899.7 billion in 2015, an annual average growth rate of 11.9%, which was the market size of $ 289.4 billion in 2005.

The anti-aging industry market in Korea is expected to show rapid growth similar to the growth of the global market. The anti-aging market, which was about 11 trillion won in 2010, will grow at an average annual rate of 9.5% It is expected. Looking at the market forecasts for anti-aging products, anti-aging drugs are expected to grow at a CAGR of 12.4% from W2.64 trillion in 2005 to 6.668 trillion won in 2015, .

In the anti-aging product market, aging prevention and health maintenance drugs account for more than 80% of the total market. The global market has increased from 115 million won in 2006 to 138 billion won in 2008, and it is expected to increase to 252 billion won in 2015 . By country, the US accounted for 34 ~ 36%, Europe 29 ~ 30%, and Japan 9 ~ 10%. Health supplements account for around 5% of the total anti-aging product market. It increased from $ 6.8 billion in 2006 to $ 7.6 billion in 2008 and is estimated to increase to $ 11.3 billion by 2015. By country and region, the US is 34 ~ 35%, Europe 31 ~ 32%, Japan 9 ~ 10%.

In Korea, the Health Functional Food Act was enacted in 2002, and a full-fledged market began to be formed in 2004. In the early days, red ginseng products and ginseng products using ginseng were mainly produced, and glucosamine, which is effective in preventing joint diseases, had a high sales. In recent years, the product has been diversified as a product for the elderly has been consumed for various age groups. Functional mainstream was antioxidant, fatigue recovery, blood circulation improvement, and cholesterol improvement, but it was classified into prostate health and eye health.

Domestic anti-aging active substance development project is the first stage, and most of the aging researches centered on the Korean Aging Society focus on inhibition of antioxidative activity as fundamental researches such as aging mechanism, and the following compounds are being researched and developed.

Matenoside compounds

- It contains diverse low-molecular physiologically active materials such as rutin, tannin, chlorogenic acid, caffeine, theobromine, and uricol, and various functions such as antioxidation, anti-cancer and Alzheimer improvement have been reported recently.

Syriacusins Compound

- Syriacus hibiscus extract inhibited HNE (human neutrophil elastase) activity.

Clitocybin Compound

- The clitosavibin compound has similar or superior antioxidative activity to catechin, a potent antioxidant. The clitosavibin compound has a specific structure as a pure natural product isolated from a native culture of mycelial mushroom mycelium and exhibits excellent antioxidative activity and is expected to be useful as a functional food material using this activity in the future.

Cyathusals Compound

- It was confirmed that Cyathus stercoreus strain, a kind of domestic native mushroom, produces strong antioxidant activity.

The following is a list of materials that are being researched and developed by companies.

1) Antioxidant products

Contains vitamin A, C, E, melatonin, estrogen, EGCG, Co-Q, lipoic acid,

Ex) Jeunesse (Italian Parmalat company, coenzyme Q10), seed oil (Teutoburger Olmuhle, coenzyme Q10) and others

2) preventive foods for geriatric diseases

Prevent skin aging, improve memory, prevent Alzheimer, improve vision / hearing deterioration, improve dental disease, improve sexual dysfunction, prevent osteoporosis / arthritis

Ex) herbal cosmetics XIANGNI, PME88 melon sod et al.

3) Anti-metabolic disease preventive food

Barley, beans, dietary fiber, phenolic compounds, enzyme inhibitors, etc.

Ex) Angiotensin (vegetable protein) (1993), Acoruka Medellabbilan Ollender of Sweden, 'An enteric preparation for obesity treatment', CJ Patdown, Dietp et al.

4) Cardiovascular disease prevention food

Prevention of hypertension at home and abroad, improvement of blood circulation, improvement of blood neutrality

Ex) Lyme, omega 3 and others

Antioxidant-related products for anti-aging as described above have been marketed, but most of the products are mainly pharmaceuticals, health functional foods, and cosmetic products. These products have a strong feeling of medicine, and they have a feeling of discomfort about the part of the elderly that they consume as patients.

Food industry In the silver industry, various health foods are representative. When you become an elderly person, your immune system weakens, digestive function decreases, and you are exposed to various diseases. However, there are many cases where health foods that are eaten for health are rather harmful to health. Apart from the damage caused by the ill-mannered elderly people whose judgment is overpowered, the damage caused by exaggerated advertising of similar health foods without proven efficacy is not negligible. Even if the health functional food proved to be effective, there is a side effect caused by abuse that I follow because I like others. It is important to remember that health functional foods are not medicines and their efficacy varies depending on their health status and nutritional intake level. In the case of elderly people, ingestion of health food due to reduced digestive power, side effects such as digestive disorders, diarrhea and abdominal pain may also occur. Therefore, the development of a food-type product that is a mixture of medicinal materials and edible materials, which are registered as safe food ingredients, will help the healthy life of the elderly.

Korean Patent No. 1422611 discloses a method for manufacturing health beverages using mushroom, mulberry leaf, camellia seed, hornbug fruit, and gingko mushroom, and Korean Patent No. 1470949 discloses a method for producing beverages containing a mixture of mushroom extract and barley extract However, the method is different from the method of producing mushroom mixed beverage using the elevation, the situation and the herbal extract of the present invention.

SUMMARY OF THE INVENTION The present invention has been made in view of the above needs, and it is an object of the present invention to provide a method for preparing a natural product having rich antioxidant and immune function, amino acid rich condition, The present invention has been accomplished by preparing a mushroom mixed beverage having a wide range of functionality and containing a large amount of amino acids without increasing odor and optimizing the production conditions such as mixing ratio.

In order to solve the above problems, the present invention provides a method for preparing a mushroom mixture, comprising: (a) preparing a mushroom mixed extract by mixing water and a conditional extract, (b) preparing water extracts of Angelica keiskei koidz., Acanthopanax senticosus, Acanthopanax senticosus, Acanthopanax senticosus, Lycopersicum blooms, and Malt extracts, respectively, followed by extraction; And (c) mixing the mushroom mixed extract prepared in the step (a) and the Angelica keiskei bark extract prepared in step (b), Sanswort oil extract, Hwanggi extract, bamboo leaf extract, licorice extract and malt extract, followed by heating The present invention also provides a method for producing a mushroom mixed beverage.

The present invention also provides a mushroom mixed drink produced by the above method.

The present invention extracts and blends the herbal medicine having the condition and the elevation, antioxidant and immunity functions under optimal conditions to make it rich in amino acid while taking advantage of the simple characteristics possessed by each material, and also has antioxidant, By developing functional beverages, consumers can further improve their health. In addition, the mushroom mixed beverage of the present invention is excellent in taste and easy to drink, and does not cause any problem even if it is used in a large amount for a long period of time without adding any food additives, thereby providing a specialized health drink for the elderly.

FIG. 1 is a diagram illustrating a production process of the hot-water extract of the present invention.
2 is a diagram illustrating a process for producing an elevated hot-water extract of the present invention.
FIG. 3 is a diagram illustrating a manufacturing process of the hot water extract of Angelica gigas Nakai of the present invention.
FIG. 4 is a diagram illustrating a process for producing a hot water extract of corn oil according to the present invention.
FIG. 5 is a diagram illustrating a process for producing the water extract of the present invention.
FIG. 6 is a diagram illustrating a process for producing the licorice extract of the present invention.
FIG. 7 is a diagram illustrating a process for producing the bamboo leaf water extract of the present invention.
8 is a diagram illustrating a process for producing the malt hot-water extract of the present invention.
FIG. 9 is a diagram illustrating a manufacturing process of the mushroom mixed beverage of the present invention.
Figure 10 shows a photograph of the mushroom mixed drink of the present invention.
11 is a graph comparing the DPPH radical scavenging ability according to the concentration of mushroom extract, herbal medicine extract, and mushroom mixed drink (health mushroom tea) of the present invention.
12 is a graph comparing LDH, AST and ALT contents of acetaminophen and the mushroom extract, herb medicine extract and mushroom mixed beverage according to the present invention.
4: Acetaminophen + Angelica gigantosa extract treatment, 5: Acetaminophen + malt extract treatment, 6: Acetaminophen + saponin extract treatment, 7: Acetaminophen + 9: Acetaminophen + Lipid Extract Treatment, 10: Acetaminophen + Hwanggi Extract Treatment, 11: Acetaminophen + Mushroom Mixed Beverage Treatment
FIG. 13 is a graph comparing the amount of NO produced according to the treatment of each sample by inducing an inflammatory reaction by treating LPS with macrophages.
1: LPS treatment, 2: LPS treatment, 3: LPS + licorice extract treatment, 4: LPS + angelica extract treatment, 5: LPS + malt extract treatment, 6: LPS + , 8: LPS + bamboo extract treatment, 9: LPS + highland extract treatment, 10: LPS + Hwanggi extract treatment, 11: LPS + mushroom mixed beverage treatment

In order to achieve the object of the present invention,

(a) preparing a mushroom mixed extract by mixing water and a conditional extract and a surface extract prepared by extracting the mushroom mixed extract;

(b) preparing water extracts of Angelica keiskei koidz., Acanthopanax senticosus, Acanthopanax senticosus, Acanthopanax senticosus, Lycopersicum blooms, and Malt extracts, respectively, followed by extraction; And

(c) mixing the mushroom mixed extract prepared in step (a) and the Angelica keiskei kotsus extract prepared in step (b), corn oil extract, Hwanggi extract, bamboo leaf extract, licorice extract and malt extract, followed by heating And a method for producing a mushroom mixed beverage.

In the method for producing a mushroom mixed beverage of the present invention, the mixing of the step (a) may be performed by mixing the conditional extract and the mushroom extract in a volume ratio of 70 to 90: 110 to 130, more preferably, The topical extract may be mixed at a volume ratio of 70: 130, 80: 120 or 90: 110. Mixing the situation and elevation at the above ratio can be made into a beverage which is easy to drink because the taste and aroma of the elevation and the elevation are well mixed and the degree of preference is excellent.

In addition, in the method for producing mushroom mixed beverage of the present invention, the conditional extract of step (a) can be extracted preferably at 70 to 80 DEG C for 80 to 100 minutes, more preferably at 75 DEG C for 90 minutes The extract can be extracted preferably at 95 to 105 캜 for 40 to 60 minutes, more preferably at 100 캜 for 50 minutes. Extracting the condition and elevation under the above extraction conditions could extract the mushroom - specific odor without deteriorating the nutrients and flavor of the mushroom and extracting it with the extract suitable for high quality beverage with improved preference.

In addition, in the method for producing mushroom mixed beverage of the present invention, the Angelica giganta extract of step (b) can be extracted preferably at 85 to 95 캜 for 40 to 60 minutes, more preferably at 90 캜 for 50 minutes The extract may be preferably extracted at 95 to 105 ° C for 90 to 110 minutes, more preferably 100 ° C for 100 minutes, and the Hwanggi extract preferably has 95 to 105 Deg.] C for 110 to 130 minutes, more preferably 120 [deg.] C for 120 minutes, and the licorice extract can be extracted preferably at 95 to 105 [deg.] C for 30 to 50 minutes, , The bark extract can be extracted at 75 to 85 ° C for 50 to 70 minutes, more preferably at 80 ° C for 60 minutes, and the malt extract can be extracted from the malt The extract preferably contains 50 to 70 < RTI ID = 0.0 > It is to be extracted, and more preferably while it is possible to extract at 90 ℃ for 60 minutes. It is preferable to extract each herb medicine separately as described above because the extraction conditions for extracting the active ingredient to the maximum and improving the preference degree are different depending on the kinds of herb medicine, and it is preferable to extract the herb medicine as the above- And its pharmacological effect was excellent, and the bitter taste unique to herb medicine was reduced, and the taste was neat because it was not stiff but it could be made into an extract suitable for beverage production.

In the method for producing a mushroom mixed beverage according to the present invention, the mixing of step (c) is preferably carried out by mixing the mushroom mixture extract, Angelica keiskei koidz. Extract, corn oil extract, Hwanggi extract, bamboo leaf extract, licorice extract, The mixture can be mixed at a volume ratio of 80 to 120: 65 to 75: 75 to 85: 25 to 35: 8 to 12: 8 to 12, more preferably mixed with mushroom extract, Angelica gigantosa extract, Extract, licorice extract and malt extract can be mixed at a volume ratio of 300: 100: 70: 80: 30: 10: 10. The combination of the ingredients and the blending ratio to produce a beverage can produce a beverage having excellent taste because the taste and flavor of the mushroom and herbal medicine are well combined.

The method for producing the mushroom mixed drink of the present invention is more specifically

(a) The conditional extracts were prepared by adding water 18 to 22 times (w / v) to each of the condition and the oily lotion, and then extracted at 70 to 105 ° C for 40 to 100 minutes. To prepare a mushroom mixed extract;

(b) 18 ~ 22 times (w / v) water was added to each of Angelica japonica, Sansui oil, Hwanggi, licorice, bamboo leaf and malt and then extracted at 75 ~ 105 ℃ for 30 ~ 130 minutes. , Bamboo leaf extract, licorice extract and malt extract; And

(c) extracting the mushroom mixed extract prepared in the step (a) and the Angelica keiskei L. extract, myrtle extract, myrtle oil extract, bergamot extract, bamboo leaf extract, licorice extract and malt extract prepared in step (b) To 75: 75 to 85: 25 to 35: 8 to 12: 8 to 12, followed by heating at a temperature of 90 to 110 ° C for 15 to 25 minutes,

More specifically,

80 to 120, or 90: 100, respectively, of the conditional extract and the extract prepared by extracting the extracts at 75 to 100 ° C for 50 to 90 minutes after adding 20 times (w / v) 110 volume ratio to prepare a mushroom mixed extract;

(b) 20 times (w / v) water was added to each of Angelica gigas Nakai, Sansui oil, Hwanggi, Licorice, Bamboo leaf and Malt, and then extracted for 40 ~ 120 minutes at 80 ~ 100 ℃. Preparing an extract, licorice extract and malt extract; And

(c) extracting the mushroom mixed extract prepared in the step (a), the Angelica keiskei kots. extract, the mungbean oil extract, the yanggi extract, the bamboo leaf extract, the licorice extract and the malt extract prepared in the step (b) in a ratio of 300: 100: 70: 80: : 10: 10, followed by heating at a temperature of 100 DEG C for 20 minutes.

In the method for producing a mushroom mixed beverage of the present invention, the taste and flavor of the mushroom and herbal extracts can be further improved by mixing the extract in the step (c) and heating under the above conditions, thereby improving the preference degree.

The present invention also provides a mushroom mixed drink produced by the above method. The mushroom mixed drink of the present invention has a reduced odor and bitter taste peculiar to mushroom and herbal medicine, and can be consumed without irritation because of the fragrance of herbs that are felt to be gentle aftertaste, It has functionalities such as protection of liver function and inflammation inhibition effect. By not adding any food additives, it can provide consumers with nutritional supplements, and can provide functional beverages that consumers can take for long-term with peace of mind .

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

1) Experimental material

The condition and elevation were purchased from Moon Sangyoung mushroom and used. Donggwi, Sansui, Hwanggi, licorice, bamboo leaf and malt were purchased from Dongbu Herbal Medicine Farming Association. The extracts were prepared by adding purified water of 20 times of the total weight ratio provided by Jangheung - gun Mushroom Industry Research Institute and hot - water extraction.

2) Material extraction

The hot water extraction was carried out by cutting the raw materials and adding 20 times of water and varying the extraction temperature and extraction time. That is, water was added at a constant ratio of the weight of each material dried and cut, and the mixture was extracted with shaking with time, centrifuged (4,000 rpm, 15 minutes), and the supernatant was used as each extract (FIGS.

3) Component analysis

A) Free sugar analysis

The free sugar component was prepared by adding distilled water to 5 mL of the sample, grinding it with a homogenizer, stirring, diluting it to 100 mL, centrifuging it at 3,000 rpm for 30 minutes, purifying it with Sep-pak C ₁₈ , The filtrate was filtered using a filter (Millipore Co., USA) and analyzed by HPLC. The analysis conditions are shown in Table 1 and the content is calculated by the external standard method.

HPLC conditions for free sugar analysis Item Analysis condition equipment Agilent Technologies 1200 Series column ZORBAX Carbohydrate
(4.6 x 150 mm) menstruum 75% acetonitrile Column temperature 35 ℃ flux 1.5 mL / min Injection capacity 20 μl

B) Analysis of constituent amino acids

The filtrate obtained by hydrolysis at 110 ° C. for 24 hours was added to 15 mL of 6N HCl solution, and the supernatant was concentrated at 50 ° C. to completely evaporate hydrochloric acid and water. Then, sodium citrate buffer The solution was filtered through a 0.45 ㎛ membrane filter, and analyzed by HPLC using AccQ · Tag method. The HPLC conditions for amino acid analysis are shown in Table 2.

HPLC conditions for amino acid analysis Item Analysis condition equipment Agilent Technologies 1200 Series column WATERS AccQ · Tag column
(3.9 x 150 mm) Column temperature 37 ℃ flux 1 mL / min Injection capacity 5 μl

C) free amino acid analysis

Free amino acid analysis was carried out by adding 25 mg of sulfosalicylic acid to 10 mL of the filtrate obtained by the same method as that of free sugars and allowed to stand at 4 ° C for 4 hours, followed by centrifugation (50000 rpm, 30 minutes) , The supernatant was filtered with a 0.45 ㎛ membrane filter, and the filtrate was used as an analytical sample. Analysis conditions were analyzed under the same conditions as the constituent amino acids.

D)? -Glucan analysis

The? -glucan analysis was performed to determine total glucan and subtract the? -glucan amount to quantify? -glucan. First, 0.1 g of the sample was put into a tube, and 1.5 mL of 37% HCl was added thereto, and the solution was decomposed for 45 minutes in a constant temperature water bath at 30 ° C. Then, 10 mL of distilled water was added to the vortex, followed by incubation at 100 ° C for 2 hours. Then, 10 mL of 2N KOH was added while cooling at room temperature, and the volume was adjusted to 100 mL with 200 mM sodium acetate buffer, followed by thorough mixing. Thereafter, 0.1 mL of exo-1,3- [beta] -glucanase plus [beta] -glucosidase dissolved in 200 mM sodium acetate buffer was added to 0.1 mL of the supernatant, 0.2 mL of acetate buffer was added to the reagent blank, 0.1 mL of D-glucose standard substance and 0.1 mL of acetate buffer were added to D-glucose standards, and the mixture was incubated at 40 ° C for 60 minutes. 3 mL of GOPOD (glucose oxidase / peroxidase mixture) was added and cultured at 40 ° C for 20 minutes. Then, absorbance was measured at 510 nm.

For α-glucan, 0.1 mL of the sample was placed in a tube, 2 mL of 2M KOH was added, and the mixture was mixed for 20 minutes. After adding 8 mL of 1.2 M sodium acetate buffer, add 0.2 mL of amyloglucosidase plus invertase, mix well, and incubate for 30 minutes in a constant-temperature water bath at 40 ° C. To 0.1 mL of the supernatant, 0.1 mL of 200 mM sodium acetate buffer and 3 mL of GOPOD were added and incubated at 40 ° C for 20 minutes and then measured at an absorbance at 510 nm.

4) Manufacture of prototype

The mixing ratios of the sample extracts used in the prototype manufacturing are shown in Tables 3 and 4 below, and the beverage manufacturing process is shown in FIG. Mixing ratio of mushrooms was higher when mixed with 70: 130, 80: 120 and 90: 110 volume ratios (Table 3).

Selection of optimal mixture ratio of elevation and condition extract for mushroom beverage production Mixing Ratio (mL) flavor color incense Comprehensive preference map situation Elevation 10 190 3.0 ± 0.8 ^c 4.7 ± 0.7 ^c 2.9 ± 1.8 ^c 2.8 ± 0.1 ^c 20 180 3.2 ± 0.8 ^c 4.9 ± 0.9 ^bc 3.2 ± 1.5 ^c 2.9 ± 0.7 ^c 30 170 3.1 ± 0.7 ^c 5.1 ± 0.5 ^bc 3.1 ± 1.9 ^c 3.2 ± 0.6 ^c 40 160 3.9 ± 0.7 ^c 5.0 ± 0.6 ^bc 4.5 ± 1.1 ^c 3.7 ± 0.2 ^c 50 150 5.5 ± 0.9 ^b 6.3 ± 0.7 ^b 4.9 ± 1.4 ^b 5.4 ± 0.7 ^b 60 140 6.5 ± 0.8 ^ab 6.2 + 0.2 ^ab 5.9 ± 1.5 ^ab 5.9 + 0.5 ^ab 70 130 6.9 ± 0.7 ^a 6.5 ± 0.4 ^a 6.4 ± 1.6 ^a 6.4 ± 0.4 ^a 80 120 6.7 ± 0.6 ^a 6.7 ± 0.5 ^a 6.3 ± 1.8 ^a 6.2 ± 0.9 ^a 90 110 6.3 ± 0.4 ^ab 6.9 ± 1.3 ^ab 6.2 ± 1.4 ^ab 6.1 ± 1.1 ^a

Mixing ratios of extracts for the production of mushroom beverage prototypes are shown in Table 4 below. Mixtures of the condition and the altitude were mixed with 70: 130 volume ratio of the extracts in the condition and elevation.

Extract blending ratio for mushroom beverage manufacturing Mixture of extract (mL) Situation + elevation Angelica Corn oil Hwanggi Bamboo leaf licorice malt A 200 200 40 10 30 10 10 B 200 200 30 20 30 10 10 C 200 200 20 30 30 10 10 D 200 200 10 40 30 10 10 E 200 150 50 50 30 10 10 F 200 150 70 30 30 10 10 G 200 150 30 70 30 10 10 H 200 100 80 70 30 10 10 I 200 100 70 80 30 10 10 J 300 150 50 50 30 10 10 K 300 150 70 30 30 10 10 L 300 150 30 70 30 10 10 M 300 150 60 40 30 10 10 N 300 150 40 60 30 10 10 O 300 100 80 70 30 10 10 P 300 100 100 50 30 10 10 Q 300 100 50 100 30 10 10 R 300 100 70 80 30 10 10 S 300 100 60 90 30 10 10

5) Prototype sensory test

The color, flavor, taste, and overall preference of the 20 panelists were analyzed by 7 - point scale method to determine the mixing ratio of raw materials for the production of highly preferred prototypes. The scoring criteria are very good; 7 points, good; 6 points, good; 5 points, average; 4 points, a little bad; 3 points, bad; 2 points, very bad; 1 point. The number of samples was changed every 2 hours and the same panel was repeated 3 times. The average score was obtained except for the highest score and the lowest score at each iteration. Statistical significance was analyzed by Duncan's multiple comparison method using the SPSS program.

6) Physiological activity of raw materials and prototype

A) Sample preparation method

5 g was added to 100 mL of purified water, and the mixture was extracted at 80 ° C for 3 hours. Then, the mixture was filtered with Whatman filter paper No. 2, The frozen and dried samples were used. Mushroom beverages were used by freeze drying.

B) DPPH free radical scavenging ability

Each sample was diluted with methanol to 0, 10, 50, 250, and 500 ㎍ / mL, and 10 μl of each was added to a 96-well plate. DPPH (2,2-diphenyl-1-picrylhydrazyl) &Lt; / RTI > 100 μl of methanol was added to the blank, reacted at room temperature for 30 minutes, and the absorbance at 518 nm was measured to determine DPPH free radical scavenging activity. The DPPH free radical scavenging ability was expressed by the following equation.

DPPH radical scavenging ability (%) = (1 - absorbance of sample addition group / absorbance of sample non-addition group) x 100

C) Hepatocyte protection and function improvement experiment

(1) Collagenase perfusion

Hepatocytes from SD (Sprague-Dawley) rats were isolated by a two-step collagenase perfusion method. To obtain primary rat hepatocytes, SD rats were anesthetized with ether and sterilized with 70% ethanol. The abdomen was opened and the right atrium was opened to allow blood to be removed. The left ventricle was filled with 21 gauge gauge syringe was perfused with 100 mL of HBSS. After removal of the blood, 100 mL of HBSS containing 100 U / mL collagenase was recirculated to allow the hepatocytes to liberate. The liver tissue was removed and 60 mL of HBSS was added in a 100 mm culture dish. 11 chopped with a blade to obtain single cells. To this collagenase suspension, the cells were washed three times with Waymouth's MB 752/1 medium (5% fetal bovine serum, 2.0 mg / mL bovine serum albumin, 10 ^-6 M dexamethasone, 10 ^-7 M Insulin, 5.32 × 10 ^-2 M L-serine, 4.09 × 10 ^-2 M L-alanine, 2.67 × 10 ^-2 M NaHCO ₃ , 100 IU / mL penicillin, 100 IU / mL streptomycin, 50 μg / mL gentamycin &Lt; / RTI > sulfate). Primary rat hepatocytes were diluted to 1 × 10 ⁵ cells / mL and attached to a 96-well plate previously coated with collagen for 4 hours, and then treated with each sample.

(2) liver function test

For liver function tests, Rat primary hepatocytes were cultured in 96-well plates at a concentration of 1 × 10 ⁵ cells / mL. After 24 hours of treatment, 50 μl of the medium was quantitatively analyzed using an LDH (Rat lactate dehydrogenase), an AST (Rat aspartate transaminase) and an ALT (Rat alanine transaminase) ELISA kit purchased from BlueGene Co. . Absorbance was measured at 450 nm using a multiplate reader (BioTek, USA). The concentrations of LDH, AST and ALT in each sample were determined by standard curves at concentrations of 0, 10, 25, 50, 100 and 250 ng / mL.

(3) Inflammation inhibition experiment induced by LPS

(A) Culture of macrophages

Raw 264.7 cells were cultured in DMEM medium supplemented with 10% FBS and 1% antibiotic at 37 ° C in a 5% CO ₂ incubator. Cells were subcultured when grown to 80-90% in culture dish and cell passage number was tested only in cells not exceeding 20 cells.

(B) Measurement of nitric oxide (NO) production

The amount of nitric oxide (NO), which is an active nitrogen species generated from RAW 264.7 cells, was measured by reacting NO ² ^- form present in the cell culture with a grease reagent. RAW 264.7 cells were seeded in 96-well plates at a concentration of 1 × 10 ⁵ cells / mL, and the samples were treated at different concentrations and cultured for 24 hours. 50 μl of each of the cell culture supernatant (100 μl) and a grease reagent (A reagent: 1% sulfanilamide and B reagent: 25% naphthylethylenediamine 0.1% in phosphate) were mixed and reacted on a 96-well plate for 10 minutes. Absorbance was measured at 540 nm using a plate reader. The NO ² ^- standard curve was prepared by concentration of NaNO ₂ .

7) Analysis of prototype components

The pH of the prototype was measured with pH meter, the sugar content was measured with sugar solution, and the total acid was titrated with 0.1N NaOH solution, and then 0.009 was multiplied by lactic acid. The lightness, lightness, redness, and yellowness of the lightness, lightness, and yellowness were measured three times with a colorimeter (COLOR JS555, USA). Brightness, redness, and yellowness of the standard white plate were 98.49, -0.02, and -0.40, respectively. Viscosities were measured using a viscometer (BROOKFIELD DV-II + Pro, rtp serial-8484417, spindle 62, brookfield engineering). Free sugars and organic acids were carried out in the same manner as above.

8) Microorganism test of prototype

Bacterial counts were measured by the plate agar culture method after the samples were collected aseptically, and cultured at 37 ° C for 48 hours using a BGLB medium and a Durham fermentation tube. Respectively.

Example 1: Analysis of ingredients of raw materials

1) Free sugar content of raw material

As shown in Table 5, the total free sugar content of raw materials was 37.41%, which was the highest in the order of 16.02%, 13.7%, 12.91%, and 5.2%, respectively. Five kinds of free sugars were detected. The main free sugars were fructose, glucose and sucrose. Trehalose was detected at 1.54% at elevation only.

Free sugar analysis of raw materials (%) Raw material Fructose Glucose Sucrose Maltose Trehalose Total Elevation 1.61 2.05 - - 1.54 5.2 Angelica - - 0.30 1.26 - 1.56 Corn oil 16.84 16.18 4.39 - - 37.41 Hwanggi 0.04 0.08 5.6 10.3 - 16.02 licorice 1.20 0.91 0.82 0.67 - 3.6 Bamboo leaf 4.6 2.5 6.6 - - 13.7 situation - - - - - 0 malt 2.33 5.20 - 5.38 - 12.91

2) Analysis of constituent amino acids

Table 6 shows the results of measuring the constituent amino acid contents of the raw materials. In total, 16 kinds of amino acids were detected and total amino acid content was 15,221.83 mg%. The total amino acid content of the situation was 9,521.29 mg%, malt 9,731.92 mg%, licorice 7,923.72 mg%, bamboo leaf 11,976.91 mg%, and angelica 10,955.91 mg%. The total amino acid content of corn oil and hwanggi was lower than the other samples.

Analysis of constituent amino acids of raw materials (mg%) amino acid Elevation Angelica Corn oil Hwanggi licorice Bamboo leaf situation malt Aspartic acid 1,224.09 1092.66 64.81 28.31 1,130.53 1,606.07 1,014.62 652.29 Serine 1,047.07 412.65 34.92 6.66 2,667.55 836.76 872.56 458.79 Glutamic acid 2,746.34 1264.36 6.27 43.47 1,305.21 1,285.88 751.68 3,215.27 Glycine 814.26 519.62 127.69 6.23 88.09 463.81 592.10 410.26 Histidine 476.62 453.14 93.5 8.80 67.00 50.84 233.27 104.71 Arginine 1,232.13 2597.61 36.54 142.53 266.28 1,036.13 610.42 213.01 Threonine 882.15 378.92 44.10 9.18 608.94 684.33 709.05 201.71 Alanine 873.43 594.81 60.21 10.33 36.94 1,039.25 668.94 453.50 Proline 863.67 412.96 55.89 - 29.93 - 617.36 1,308.72 Tyrosine 48.31 113.82 12.69 7.17 22.93 1,068.15 226.81 362.81 Balin 812.02 934.21 60.93 7.05 - 503.74 667.63 403.13 Methionine 234.23 204.62 12.03 2.05 4.06 160.32 101.20 116.16 Lysine 1,487.27 601.36 79.20 14.86 89.03 928.73 590.80 272.06 Isoleucine 721.64 362.63 55.55 5.72 44.92 485.28 563.68 231.81 Leucine 1,193.86 647.62 116.01 12.77 13.41 963.19 933.16 695.38 Phenylalanine 564.74 364.92 42.37 10.29 1,548.90 864.43 368.01 632.31 total 15,221.83 10,955.91 902.71 315.42 7,923.72 11,976.91 9,521.29 9,731.92

3) Free amino acid analysis of raw materials

The free amino acid content of the raw materials was measured as shown in Table 7, and a total of 16 kinds of amino acids were detected on the surface, and the total free amino acid content was 4,012.68 mg%. The total free amino acid content of the situation was 1,894.35 mg%, malt 1,850.19 mg%, licorice 1,855.67 mg%, bamboo leaves 2,276.99 mg% and angelica 2,627.31 mg%. Compared with the constituent amino acid contents, all of the sample contents were lowered. In essential oils, three essential amino acids (valine, isoleucine, leucine) and two non - essential amino acids (tyrosine, phenyl alanine) were not detected.

Free amino acid analysis results of raw materials amino acid Elevation Angelica Corn oil Hwanggi licorice Bamboo leaf situation malt Aspartic acid 365.03 262.03 0.84 6.63 264.76 305.34 258.45 124.01 Serine 288.07 98.96 0.28 1.56 624.72 159.08 113.77 87.22 Glutamic acid 689.25 303.20 0.32 10.18 305.67 244.46 167.75 611.27 Glycine 220.61 124.61 0.08 1.46 20.63 88.18 109.74 78.00 Histidine 138.80 108.67 1.31 2.06 15.69 9.67 83.73 19.91 Arginine 313.87 622.93 2.04 33.38 62.36 196.98 96.51 40.50 Threonine 248.65 90.87 0.62 2.15 142.61 130.10 150.54 38.35 Alanine 149.57 142.64 0.16 2.42 8.65 197.58 139.91 86.22 Proline 209.37 99.03 0.10 - 7.01 - 79.33 248.81 Tyrosine 11.49 27.29 - 1.68 5.37 203.07 51.38 68.98 Balin 228.63 224.03 - 1.65 - 95.77 138.13 76.64 Methionine 64.50 49.07 0.50 0.48 0.95 30.48 36.57 22.08 Lysine 370.03 144.21 1.32 3.48 20.85 176.56 71.34 51.72 Isoleucine 182.69 86.96 - 1.34 10.52 92.26 43.15 44.07 Leucine 374.40 155.30 - 2.99 3.14 183.12 162.88 132.20 Phenylalanine 157.72 87.51 - 2.41 362.74 164.34 191.17 120.21 Sum 4,012.68 2,627.31 7.57 73.87 1,855.67 2,276.99 1,894.35 1,850.19

4) Raw material β - glucan analysis

The content of β-glucan in the raw materials was as shown in Table 8, and an elevation of 37.15%, a condition of 19.64%, and malt of 4.62% were detected, but the remaining samples were not detected.

Β - Glucan analysis of raw materials division Elevation Angelica Corn oil Hwanggi licorice Bamboo leaf situation malt β - glucan 37.15 - - - - - 19.64 4.62

Example 2: Sensory evaluation of mushroom mixed drink

The sensory evaluation showed that the R test was the best in flavor and taste, and the color was excellent in Q test. Compared to the other test groups, the R test group showed the most significant difference in overall acceptability (Table 9). As a result of sensory evaluation, the preference of mushroom beverage had the greatest effect on the addition of the extracts and the addition of the extract of the mushroom extract. As a result, the R test group with the highest sensory test results was selected as the final blend ratio.

Sensory test Item incense flavor color Comprehensive preference map A 3.3 ± 1.1 ^d 3.3 ± 0.8 ^d 4.0 ± 0.9 ^bc 3.6 ± 0.7 ^c B 3.0 ± 0.9 ^d 2.6 ± 1.0 ^d 3.7 ± 1.0 ^d 2.9 ± 0.7 ^d C 3.6 ± 0.8 ^d 3.0 ± 0.8 ^d 3.8 ± 1.1 ^cd 3.4 ± 0.7 ^d D 2.9 ± 0.8 ^d 2.7 ± 1.2 ^d 3.6 ± 1.0 ^d 3.2 ± 0.6 ^d E 3.0 ± 0.6 ^d 3.1 ± 0.9 ^d 3.7 ± 1.0 ^d 4.5 ± 0.7 ^bc F 3.2 ± 1.1 ^d 3.4 ± 1.0 ^d 4.1 ± 1.0 ^cd 3.6 ± 0.5 ^cd G 2.9 ± 1.0 ^d 3.4 ± 1.1 ^d 3.6 ± 1.0 ^d 4.1 ± 0.9 ^c H 3.6 ± 0.9 ^d 3.5 ± 0.9 ^d 3.2 ± 1.2 ^d 4.6 ± 0.9 ^c I 4.2 ± 1.4 ^cd 3.7 ± 1.3 ^d 3.9 ± 1.1 ^d 4.9 ± 1.3 ^c J 4.6 ± 0.9 ^c 4.1 ± 1.3 ^c 3.6 ± 1.0 ^d 4.8 ± 1.4 ^c K 4.3 ± 0.3 ^cd 4.1 ± 0.1 ^cd 4.2 ± 0.4 ^cd 4.2 ± 0.3 ^cd L 4.7 ± 0.4 ^c 3.9 ± 0.4 ^d 4.8 ± 0.5 ^c 4.8 ± 0.4 ^c M 3.9 ± 0.8 ^d 5.0 ± 0.8 ^c 4.7 ± 0.2 ^c 4.7 ± 0.4 ^c N 4.8 ± 0.6 ^c 5.1 ± 0.06 ^c 4.9 ± 0.6 ^c 4.9 ± 0.5 ^c O 5.0 ± 0.06 ^c 5.0 ± 0.3 ^c 4.6 ± 0.4 ^c 5.1 ± 0.11 ^b P 5.0 ± 0.2 ^b 5.3 ± 0.1 ^b 5.7 ± 0.3 ^b 5.9 ± 0.3 ^ab Q 5.4 ± 0.7 ^ab 5.8 ± 0.5 ^ab 6.5 ± 0.6 ^ab 6.1 ± 0.3 ^ab R 6.6 ± 0.5 ^a 6.2 ± 0.4 ^a 6.3 ± 0.4 ^a 6.5 ± 0.2 ^a S 5.3 ± 0.3 ^ab 5.9 ± 0.1 ^ab 6.3 ± 0.2 ^ab 6.0 ± 0.3 ^ab

Example 3: Analysis of the components of the prototype

Table 10 shows the analysis results of the R test strip, which is a prototype of mushroom drink. The prototype had a pH of 5.32 and a chromaticity of 70.93, a (redness) value of 2.34 and b (yellowness) value of 41.24. The prototype was transparent and yellowish. The sugar content was found to be 5.4 brix. The viscosity was measured at 25 rpm at 20 rpm and the CP value was 2.9.

Analysis of the components of the prototype division pH Chromaticity Sugar content
(brix) Viscosity Total
(%) Prototype 5.32 L 70.93 5.4 rpm 21 0.63 a 2.34 ℃ 26 b 41.24 cP 2.9

Example 4: Physiological activity of prototype

1) DPPH free radical scavenging ability

The free radical scavenging activity of DPPH radical scavenging activity of vitamin C, the situation, the BHT, and the concentration of the alcoholic beverages (0, 10, 50, 250, 500 ㎍ / mL) The elimination ability was measured in the order of corn oil, mugwort, angelica, licorice, mushroom drink, malt, mulberry, and hwanggi (Fig. 11). All of the extracts were lower than the vitamin C, which is a positive control, but the extracts were higher than the edible antioxidant BHT. And mushroom beverage showed low DPPH radical scavenging ability, but it showed about 18% scavenging ability at 500 ㎍ / mL concentration.

2) hepatic function-protecting efficacy against acetaminophen

The high concentration of acetaminophen caused toxicity in hepatocytes. As a result of confirming the content of LDH (A in FIG. 12), the content of AST (FIG. 12B) and the content of ALT (FIG. 12C) Extracts, extracts of mushroom, mushroom beverages, extracts of hwanggi, bamboo shoots, and licorice extract.

3) Measurement of NO (Nitric oxide) production

Raw 264.7 macrophages were treated with 1 ㎍ / mL LPS to induce an inflammatory response. Each sample was treated with 100 ㎍ / mL (Fig. 13) And the decrease of inflammatory reaction in the order of extracts.

4) Mushroom beverage microorganism inspection

Table 11 shows the results of the general bacterial test and E. coli test according to the treatment temperature and time after mixing the extracts. General bacteria and Escherichia coli did not occur, and it was found to be microbiologically stable during storage. Therefore, ingestion of this beverage can be proved to be microbiologically safe.

Microbial test result of mushroom drink microbe Control Heating temperature (占 폚), time (minute) 60 ° C 80 ℃ 100 ℃ 10 minutes 15 minutes 20 minutes 30 minutes 40 minutes 5 minutes 10 minutes 15 minutes 20 minutes 30 minutes 5 minutes 10 minutes 20 minutes Normal
Germ - - - - - - - - - - - - - - Escherichia coli - - - - - - - - - - - - - -

Claims

(a) 20 times (w / v) of water was added to each of the conditions and elevation, and the conditions were 80 to 100 minutes at 70 to 80 ° C, and 95 to 105 ° C for 40 to 60 minutes, respectively Preparing a mushroom mixed extract by mixing the conditional extract and the altitude extract at a volume ratio of 70: 130, 80: 120 or 90: 110;
(b) After adding water 20 times (w / v) to each of Angelica gigas Nakai, Sansui oil, Hwanggi, Licorice, Liliaceae and Malt, 40 ~ 60 minutes at 85 ~ 95 ℃, 90 ~ 110 to 130 minutes at 95 to 105 ° C for Hwanggi, 30 to 50 minutes at 95 to 105 ° C for licorice, 50 to 70 minutes at 75 to 85 ° C for bracts, 50 to 70 minutes at 85 to 95 ° C Extracts of Angelica keiskei koidz, Extract of Cornus syrup, Hwanggi extract, Bamboo leaf extract, Licorice extract and malt extract; And
(c) extracting the mushroom mixed extract prepared in the step (a), the Angelica keiskei kots. extract, the mungbean oil extract, the yanggi extract, the bamboo leaf extract, the licorice extract and the malt extract prepared in the step (b) in a ratio of 300: 100: 70: 80: : &Lt; / RTI > 10: 10, and then heating the mixture at a temperature of 100 DEG C for 20 minutes.

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