KR100615039B1 - Method of oriental herbal yogurt - Google Patents

Abstract

The present invention relates to a method for producing a functional herbal yogurt for hangover and liver protection, the raw materials of herbal and mushroom materials used are hawthorn, walnut fruit, alder, ganoderma lucidum mushroom, shiitake mushroom, extracted from these mixtures Sterilized by adding skim milk powder and sugar to the extract, and then cooled to room temperature, and mixed with a pre-cultivated mixed lactobacillus starter. The lactobacillus was incubated at 37 ± 1 ℃, and the milk casein protein was cured by the acid produced by the lactobacillus. It relates to a method of producing a functional herbal yogurt, characterized in that the fermentation is made to the time when it is completely formed.

Functional herbal yogurt according to the present invention is obtained by mixing the mushroom material ganoderma lucidum mushroom and shiitake mushrooms with excellent protection of liver function in addition to the herbal material material hawthorn, barberry, alder, which has a hangover effect by lowering blood alcohol concentration By using water-soluble extracts, it lowers blood alcohol and acetaldehyde levels due to alcohol intake, promotes alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activity of alcohol and acetaldehyde degrading enzymes in liver tissue, liver function It can be seen that the clinical indicators of serum AST and ALT activity, hepatic tissue neutral lipid concentration decrease, oxidative stress inhibitory effect by decreased lipid peroxidation concentration, alcoholic hepatotoxicity and fat degeneration effect by histological observation.

In addition, the functional herbal yogurt according to the present invention is more effective against alcoholic liver toxicity by adding mushrooms in addition to medicinal herbs, barberry and alder, as well as hepatotoxicity due to alcohol intake, and especially high incidence in Korean men. It will also contribute to improving or preventing the disease. In particular, the extract obtained by the appropriate mixing ratio of herbal and mushroom materials significantly increases the number of lactic acid bacteria in fermented milk during fermentation process, and by ingesting them, the greatest advantages that lactic acid bacteria can have, intestinal environment improvement, immunity enhancement, allergy reduction, prevention of tooth decay, In addition to various effects, such as cholesterol reduction, it can be expected to have an additional effect of promoting metabolism due to the intake of the vitamin B group produced during fermentation.

Functional, Chinese herbal medicine, yogurt, hut tree, hut fruit, alder tree, ganoderma lucidum mushroom, shiitake mushroom, skim milk powder, lactic acid bacteria, starter, hangover

Description

Method of manufacturing functional herbal yogurt {Method of Oriental herbal yogurt}

       1 is a graph showing the results of weight change over time by the administration of functional herbal yogurt in rats administered with alcohol as an embodiment according to the present invention.

      Figure 2 is a graph showing the results of changes in beverage intake over time by administration of functional herbal yogurt in alcohol-administered rats.

      Figure 3 is a graph showing the results of serum triglyceride concentration by functional herbal yogurt administration of alcohol-administered rats as an example according to the present invention.

      Figure 4 is a graph showing the results of the activity of alcohol degrading enzymes and acetaldehyde degrading enzymes in liver tissue by functional herbal yogurt administration of alcohol-administered rats according to an embodiment of the present invention.

      Figure 5 is a graph showing the results of serum ALT, AST, LDH activity showing the indicator of liver tissue toxicity by functional herbal yogurt administration of alcohol-administered rats according to an embodiment of the present invention.

      Figure 6 is a graph showing the results of the concentration of serum alcohol and acetaldehyde by functional herbal yogurt administration of alcohol-administered rats according to an embodiment of the present invention.

      Figure 7 is a graph showing the results of changes in lipid peroxide concentration of liver tissue fractions by functional herbal yogurt administration of alcohol-administered rats according to an embodiment of the present invention.

The present invention relates to a method for producing functional herbal yogurt, and more particularly, mushroom material having excellent protective effect of liver function along with hut tree, hut fruit, alder, which are herbal materials which have hangover effect by lowering blood alcohol concentration. After mixing phosphorus ganoderma lucidum mushroom and shiitake mushrooms at an appropriate ratio and extracting them under conditions of high temperature and high pressure, the water-soluble extracts are fermented with lactic acid bacteria and consumed by those who drink them, which helps to relieve hangovers and protects liver function and improves health. It relates to a method for producing a functional herbal yogurt characterized in that it can be further improved.

With the recent economic growth, the increase in the adult obese population due to dietary changes, the stress caused by heavy work, excessive drinking or various diseases, the number of patients with liver disease, including fatty liver continues to increase. In particular, Koreans have a higher rate of liver disease, including hepatitis, than other foreigners due to the traditional habits of dietary culture, and the mortality rate from liver cancer is the world's No. 1 at 23.4 per 100,000 people, and chronic liver disease is 28.8. Statistics have already been reported that the number of patients with liver disease is high enough to occupy the stomach.

 In particular, in modern times, frequent drinking spots to relieve the stress caused by heavy work causes chronic fatigue, and after excessive drinking, acetaldehyde produced by metabolism of alcohol in the body does not decompose into carbon dioxide and water, but moves around in the blood. Hangovers such as vomiting, thirst, headaches, or decreased appetite continue until the day after drinking, and the consumption of drinking hangover drinks before and after drinking is increasing rapidly in recent years.

In order to solve the hangover, acetaldehyde, which is an alcohol metabolite, does not remain in the blood, but alcohol absorbed in the body does not remain in the blood. The cause is eliminated, this hangover takes a certain amount of time depending on the type and amount of alcohol consumed and the physical condition of the person. After all, the hangover is to improve the efficiency of detoxification and excretion. Therefore, by constantly ingesting functional ingredients that can promote liver function to prevent the rapid liver toxicity caused by drinking in advance, health functional foods ingested to prevent hangover and protect the liver before and after drinking It is getting a lot of attention, and various products related to this are being developed and marketed, and the drink beverage for hangover is leading. The market for hangover drink beverages is in the spotlight mainly because male workers in their 30s and 50s are in line with the drinking class of liquors who prefer relatively high alcohol such as soju and whiskey. There is a growing trend.

Various hangover drinks that have been developed for the above purpose are prepared by adding a small amount of compounds extracted from fungi such as plants or ganoderma lucidum and situation mushrooms, such as lianas and alders.

Therefore, looking at the patent application as a technology that is used as a dietary supplement for health hangover and hangover removal using the compounds extracted from plants and fungi as described above, Republic of Korea Patent Publication No. 2002-0345798 (2002. 07. 24) technology for manufacturing hangover-relieving functional foods made by extracting and concentrating the earth, alder, and sorghum using a solvent mixed with water and ethyl alcohol in a certain ratio and Korean Patent Publication Technology for manufacturing natural tea for hangover elimination using the main raw material of directly powdered leaves, stems and roots of alder and rowan of 1998-0181168 (announced on Feb. 01, 1999) and Korean Patent Publication No. 2003 -0385844 (Notice of 2003. 06. 02) Production of hangover-relieving beverages, which are easy to take and fast absorbed by the human body, using the ferns (earths), leaves, and berries as main ingredients. And technology for manufacturing hangover-resolved beverages after drying the bark trees (earths), leaves, and fruits of Korea Patent Publication No. 2003-0385844 (announced on May 19, 2003) and drying water. Health supplements for hangover after drinking alcohol in 2002- 0023770 (published on March 29, 2002), on the manufacture of natural round, taburette, and wexel forms mainly of the bark, snow, red nourishment The main ingredients are the extracts of Korean native hawthorn trees (leaves, stems and fruits) and alder of Korea Patent Publication No. 2002-0048212 (published on June 22, 2002) and put herbal extracts such as brown flowers and taurine There are known techniques for the manufacture of hangover-relieving beverages, but in the case of the above patents, the components extracted or concentrated from the leaves and berries of the lianas and alders or their leaves and berries, which are known to be mainly effective for hangover, and many other parts. Although the prepared by simply adding the herbal medicines or components having a functionality that was not fully demonstrate the effect of eliminating hangover by promoting mutual action of these additional component problem.

On the other hand, fermented milk products fermented by inoculating lactobacillus with milk or skim milk powder as raw materials have an active ingredient such as lactic acid, peptone, peptide and oligosaccharide produced by lactic acid bacteria during the fermentation process. It is recognized as a representative health drink that is closely related to health. In recent years, functional fermented milk that emphasizes health orientation has been researched and developed to provide specific functionalities directly related to lifestyle-related diseases such as functional products related to the digestive system, products that meet human desires for beauty, and products for lowering blood sugar levels of diabetics. Products with palatability are on the market, and in fact, the production of these products is mainstream, which is revealed in recent statistics.

 However, the commercially available hangover-resolved fermented milk uses high-density sweeteners such as sugar, fructose, apple pear concentrate, red grape concentrate, and bokbunja concentrate juice, and raw milk containing large amounts of fat. As well as having a problem, conventional fermented milk made for hangover elimination has been prepared by adding concentrated extracts of holly or alder trees, mainly using raw milk as the method of preparing hangover-eliminating beverages. By using a mixture of these bark and alder, rather than using only the barn or alder such as this alone, to have a synergistic effect of relieving hangover, and also to produce fat that can cause obesity The product which can make the most of the minimum degreasing oil which can be made The foot is desperately required.

In addition, in order to sufficiently extract beneficial ingredients from various herbal ingredients that are effective in relieving hangovers, the extract should be extracted twice in the same manner as the herbal medicine extract and it should be used as a lactic acid bacteria culture source by adding skim milk powder to it. However, in existing commercial products, it is inevitable to concentrate the extract of herbal ingredients due to the limited amount of extract added due to the use of raw milk, which may lead to unnecessary expense expenditure for this as well as change in the process. In addition, it is possible to diversify Korean herbal medicines with excellent liver function protection as well as to relieve hangovers. However, due to the limited amount of extracts used, only one of the components extracted from liana or alder tree is selected and used. In order to solve this problem, a huge foreign currency outflow has been pointed out as a problem by importing and adding milk thistle complex extract, lecithin (soy), vitamins, chicory oligosaccharides, etc. from abroad.

     The present invention is to solve the problems as described above is effective in protecting the liver function and herbal function mixed with ingredients extracted from the bark, barberry, alder tree which is effective in resolving hangover by the decomposition of alcohol and acetaldehyde in blood By mixing mushroom materials mixed with ingredients extracted from Ganoderma lucidum and dried shiitake mushrooms at an appropriate ratio, the mixed extract is prepared by adding fermented yogurt to the fermented yogurt, adding an additional effect on the hangover and liver protection. The present invention has been completed by producing a functional herbal yogurt characterized in that.

The effect of the hangover resolution of the components extracted from the plants and fungi according to the present invention was increased by 36 to 42% in the extract intake group of the fruit of the hawthorn fruit compared to the 28% increase in the control group 1 hour after alcohol intake in animal experiments It has been found that blood alcohol levels are reduced (Kim MH, Chung YT, Lee JH, Park YS, Shin MK, Kim HS, Kim DH, Lee HY. 2000. Hepatic detoxification activity and reduction of serum alcohol concentrations of Hovenia dulcis Thunb from Korea and China. J-Medicinal Crop Sci 8, 225-233) and acetaldehyde, the causative agent of hangover, is a metabolite of ADH action, and if the fruit extract simply activates the enzyme, the alcohol in the blood rapidly decreases, but acetaldehyde in the liver and blood. It is more likely to accumulate and cause hangover symptoms, and ADH and ALDH enzyme activity was almost similar to 28.8% and 29.8% in the alcohol group, but ALDH activity was activated by 10% more than ADH activity in the fruit extract group. Acetaldehyde was decomposed, and Ahn et al. (Ahn et al.) Showed a single alcohol toxicity test of ethanol, alder, and mixed extracts that are effective in relieving hangover. It has been found to increase up to 60% in the mixed wood extract (An SW, Kim YG, Kim MH, Lee BI, Lee SH, Kwon HI, Hwang B, Lee) HY.1999.Comparison of hepatic detoxification activity and reducing serum alcohol concentration of Hovenia dulsis Thung and Alnus japonica Steud. Korean J Medicinal Crop Sci 7, 263-268). In addition, the time taken for 20 adult men to decrease the alcohol concentration from 0.182% to less than 0.004% after 15 minutes of alcohol intake was 6 hours of alder, 5 hours of larvae, and 4 hours of mixed extracts of sheds and alders. The acetaldehyde concentrations associated with hangovers were degraded more quickly in the mixed extracts than in the fern and alder trees alone. These findings suggest that the use of a combination of hut and alder trees, which are effective for hangover, is more effective.

In addition, the extract of the hawthorn and alder tree has the effect of promoting the decomposition of alcohol faster than non-drinking by promoting the enzyme activity involved in alcohol degradation in the liver. If a person consumes alcohol on a daily basis, it is possible to think that there is a high possibility that the function of alcohol degrading enzyme activity and liver function may be reduced due to overuse. To solve this problem, it is necessary to increase the supply of high-quality protein and liver protection, which are the main raw materials for the production of alcohol degrading enzymes. In particular, mushrooms such as Ganoderma lucidum and shiitake mushrooms have been shown to be effective in promoting liver function and have been published in several papers and are used as raw materials for the treatment of liver diseases. This can be a good source of protein.

Therefore, the present invention is prepared by adding to the yogurt fermented with lactic acid bacteria using the above-mentioned extraction ingredients to lower blood alcohol concentration and acetaldehyde concentration due to alcohol intake, alcohol dehydrogenase which is alcohol and acetaldehyde degrading enzyme of liver tissue ( ADH) and acetaldehyde dehydrogenase (ALDH) activity promotion, serum AST and ALT activity, clinical markers of liver function, hepatic tissue neutral neutral lipid concentration, oxidative stress inhibitory effect by decreased lipid peroxidation concentration, histomorphologic observation It is an object of the present invention to provide a method for producing a functional herbal yoghurt, characterized by reducing alcoholic liver toxicity and fatty degeneration.

Still another object of the present invention is to distinguish male and female by preference food by adding the extracting ingredients as described above to the yogurt fermented with lactic acid bacteria skim milk that can minimize the fat that can cause obesity It is a multifunctional food that can be prevented from aging after drinking, as well as improving gastrointestinal function, preventing blood vessel disease by reducing blood lipid concentration and antioxidant activity by decreasing lipid peroxidation concentration. It is to provide a method of manufacturing herbal yogurt.

       Hereinafter, the manufacturing method of the present invention for achieving the above object will be described in detail.

Raw materials of herbal and mushroom materials used in the manufacture of functional herbal yogurt for hangover relief and liver protection according to the present invention are hawthorn, walnut fruit, alder tree, ganoderma lucidum mushroom, shiitake mushroom, and the skim milk powder in the extract extracted from these mixtures. Sterilize by adding sugar, cool to room temperature, add pre-cultured mixed lactobacillus starter, and incubate the lactic acid bacteria at 37 ± 1 ℃ to completely form curd by acid produced by lactic acid bacteria. It is prepared by fermentation.

The herbal material that can be used in the present invention is preferably used to produce herbal yogurt by mixing with other mushroom material extracts using the extracts of the barberry, barberry and alder which have hangover relief and liver function protection effects. It is not limited to various kinds of extracts, and all food and medicinal herbs that have a hangover relief and liver function protection effect can be used, and the production place is not important.

And using the extract of shiitake mushrooms and ganoderma lucidum which can be used in the present invention, it is preferable to prepare a functional herbal yogurt by mixing with other herbal extracts, but is not necessarily limited to the extract of the type described above, food, medicinal Possible, all food, medicinal mushrooms that can synergize by complementing the hangover resolution function of the herbal extracts can be used.

Looking at the manufacturing method of functional herbal yogurt according to the present invention.

Ⅰ) Thinly cut dried bark and alder into 94.0 ~ 99.8% by weight of water, add 0.1 ~ 2.0% by weight and 0.1 ~ 2.0% by weight of barberry, and boil for 30 minutes to 2 hours at high temperature of 98 ~ 121 ℃. Obtaining an extract of the components contained in the tree and the fruit (medicinal herb component extraction step);

Ii) chopped dried shiitake mushrooms and dried ganoderma lucidum in 99.0 ~ 99.5% by weight of water extracted from boiled barn tree, barberry, alder in step iii) and cut the size of horizontal and vertical 0.5 ~ 1 cm Putting 0.05 ~ 1.0% by weight shiitake mushrooms under the same extraction conditions as in step iii) to extract the ingredients contained in Ganoderma lucidum and shiitake mushrooms (mushroom ingredient extraction step);

Iii) preparing a lactic acid bacterium fermentation broth by mixing 7-15% by weight of skim milk powder (based on solids) with 85 to 97% by weight of the extract extracted from the herbal and mushroom components prepared through steps iii) and ii) step);

Iii) pre-culturing the mixed lactic acid strain in the lactic acid bacteria culture medium which has been sterilized separately at 80 ° C. to 100 ° C. (mixed lactic acid strain culture step);

Iii) 1 ~ 3% by weight of the starter (starter) pre-cultured the lactic acid strain mixed in step iii) to 97 ~ 99% by weight of the lactic acid bacteria fermentation broth prepared in step iii) to about 8 ~ at 37 ± 1 ℃ Incubation for 12 hours (lactic acid bacteria fermentation step);

Finally, functional herbal yogurt is prepared.

Barnwood extracted from the step ⅰ) ( Hovenia dulcis Thunb) contains many biologically active ingredients, such as (+)-ampelopsin, laricetin, myricetin, and (+)-gallocatechin. Flavonoids (flavonoids), flavonols (hovenitin I, II, III, etc.), peptide alkaloids (frangulanine) is identified. Biological activity has been reported to inhibit liver toxicity, histamine release, antioxidant, antibacterial, alcoholic muscle relaxation.

The fruit of the barn tree is known as the earth and is used for the treatment of alcohol, anemia, gugal, vomiting, sajima rain, and fecal pain. In the main herbaceous tree (Kim Il-sik, 1992), the fruit of the barn tree is brown with a faint scent. It is said to have a sweet taste to eat and drink alcohol, and the juice is said to loosen the poison and stop nausea.

The alder tree contains several flavonoids and tannins in its bark, and the fruit contains more than 10% tannins and about 0.2% essential oils. In oriental medicine, the bark of alder is called yangyang, and it has been known to be used for fever, hemostasis, convergence, etc. In recent years, alcohol resolution and liver detoxification have been discovered along with the bark tree, and it has been added to some hangover drinks. (An SW, Kim YG, Kim MH, Lee BI, Lee SH, Kwon HI, Hwang B, Lee HY.1999.Comparison of hepatic detoxification activity and reducing serum alcohol concentration of Hovenia dulsis Thung and Alnus japonica Steud. Korean J Medicinal Crop Sci 7, 263-268)

In the above, the addition amount of the barberry and alder is 0.1 to 2.0% by weight, the addition amount of the barberry is preferably 0.1 to 2.0% by weight. If the addition amount of the bark, alder, and barberry is less than the above added amount, there is a fear that the alcohol resolution and liver detoxification action may be lowered. There is a possibility that the effect of yogurt will not be raised and the taste of yogurt will be lost.

And the amount of the hut and alder to add to the water in the above is a part of the alder as a function to assist the alcohol resolution and liver detoxification of the extract component of the hut, so the mixed amount of the hut and alder is 75 ~ 85% to 15-25% by weight is preferred.

In the above extraction, the extraction may be repeated once or twice and filtered using gauze.

The Ganoderma lucidum and shiitake mushroom extracts extracted in step ii) are added to the lactic acid bacteria culture medium, so that the mixed strains used in the production of the functional herbal yogurt of the present invention are Lactobacillus ashdophyllus ( L. acidophilus ) and Lactobacillus thermophilus. ( L. thermophilus ) and Bifidobacterium loggum have been reported to not only promote the growth of Bifidobacterium loggum , but also to increase the intestinal lactic acid bacteria by ingesting mushroom extract. (Han ML, Bae EA, Rhee YK , Kim DH. 1996. Effect of mushrooms on the growth of intestinal lactic acid bacteria.Korean J Food Sci Technol 28, 947-952). At this time, as the amount of Ganoderma lucidum extract increased, there was a clear effect on the growth of Lactobacillus thermophilus, and lysine, serine, arginine, and glutamic acid were added among the free amino acids. The growth of Lactobacillus thermophilus was also promoted (Koo HH, Chung SH. 1994. Effects of Panax ginseng and Ganodema lucidum extract on the growth of lactic acid baxteria. Korean J Food Nutr 7, 45-50).

In addition, functional herbal yogurt suppresses the growth of harmful bacteria by improving the intestinal environment of lactic acid bacteria by increasing the number of lactic acid bacteria by extracting herbal and mushroom materials, supplying abundant vitamin B group, promoting intestinal mineral absorption, reducing blood cholesterol, immune function Bioactive effects such as hyperactivity can also be expected.

In addition to the Ganoderma lucidum and shiitake mushrooms, edible and medicinal mushrooms such as situation mushrooms and agaricus are known as mushrooms that are expected to have the physiological activity as described above.

In the above, the amount of dried shiitake mushrooms and dried ganoderma lucidum added to boiled bark, barberry, alder and extracted water is preferably 0.05 to 1.0% by weight, and when the amount is less than the above range, the anticancer effect of the physiological activity of the mushrooms. , Mutagenicity inhibitory effect, serum lipid concentration lowering effect, immuno-enhancing effect, etc., may be lowered. If the amount exceeds the above-mentioned amount, the physiological activity of mushrooms does not increase with the increase of the amount of addition, and the taste of yogurt is rather increased. It may be lost. The shiitake mushrooms and dried ganoderma lucidum are additional components in addition to hangover elimination, but the amount of the mixture is not particularly limited, but 40 to 60% by weight to 40 to 60% by weight is mixed to exhibit additional functions. It is preferable to make it.

The powdered milk used in step iii) may be used by using skim milk powder alone or by mixing a small amount of skim milk with skim milk powder. In this case, the amount of the skim milk powder used is preferably 7 to 15% by weight (based on the total solid content), because the extract of the herbal and mushroom components prepared through the steps iii) and ii) and the range of the amount This is because when mixed with skim milk powder, lactic acid bacteria culture medium suitable for fermenting the most lactic acid bacteria can be prepared.

And, the mixture was added to the mixed oil 97.0 - 98.95% by weight of one hut for wood alder extract and 0.1 to 2.0% by weight and 0.05 to 1.0% by weight extract of Ganoderma mushroom and the heotgae fruit. In this case, the extracts may have a decrease in alcohol resolution and liver detoxification when the amount is less than the addition amount. When the extract amounts exceed the range, the effects of alcohol resolution and hepatic detoxification are increased by increasing the amount. Rather, there is a risk that the flavor inherent in yogurt may be lost.

The ⅳ) In step Lactobacillus lactic acid bacteria culture fluid Ash's also a filler (Lactobacillus acidophilus ) , Lactobacillus Thermo's filler (Lactobacillus thermophilus), Bifidobacterium Longgum ( Bifidobacterium By mixing and culturing the strain of loggum ) , the fermentation effect of the lactic acid bacteria is enhanced by the interaction of the strains.

And in the step iii) it is preferable to add 1-3% by weight of the starter (starter) pre-cultured the mixed strain to 97 ~ 99% by weight milk powder. When the starter is added below the amount of the added amount, the powdered milk may not be fermented sufficiently. When the amount of the starter is exceeded, the milk powder may be overfermented, resulting in the loss of flavor and aroma of the yoghurt. In addition, the lactic acid bacteria is most preferably a fermentation temperature of 37 ± 1 ℃.

Functional herbal yogurt of the present invention is necessarily wasted two trees, alders, heotgae fruit and estates of mushrooms and protective efficacy ingredient beta inter addition of shiitake mushroom extract, lecithin, Fermented Turmeric, Artichoke extract, aspartic limited by the , Vitamin C, taurine, high-purity chitosan, rice snow fermented extract (Kurume), honey, acerola, tocopherol, royal jelly, ginseng, ogapi, green tea, grapefruit seed, goji berry, cheongung, brown root, eochocho, licorice extract It may be added up to 2.0% by weight based on the total weight. In addition, it is possible to enhance palatability by mixing additives such as apple concentrate, white grape and red grape concentrate, pear concentrate, bokbunja concentrate.

Hereinafter, the technical configuration and operation of the present invention will be described in detail with reference to Examples, but the scope of the present invention is not necessarily limited to these Examples.

Example  1. Manufacture of functional herbal yogurt for hangover and liver protection

1. Preparation of Functional Herbal Yogurt

Add dried shiitake, alder and walnut fruit with dried shiitake mushrooms and ganoderma lucidum cut to a size of 0.5 ~ 1 cm, and add the same amounts as described in [Table 1] below to each water. Extracted by repeated extraction once or twice at 30 ~ 2 hours at a high temperature of ~ 121 ℃ filtered by gauze. Then, the extract is added to the skim milk (total solid content 8% by weight) mixed with skim milk powder together with 5% by weight of sugar to prepare a lactic acid bacterium fermentation broth. In addition, L. acidophilus , L. thermophilus , and Bifidobacterium loggum were pre-cultured in lactic acid bacteria culture medium that had been sterilized at 80 ° C to 100 ° C. 2 wt% of mixed starter lactic acid strains were incubated at 37 ± 1 ° C. for 10 hours to finally prepare functional herbal yogurt.

And the yoghurt-free yogurt containing no extract was prepared as a control.

2. Polyphenol Content, Soluble Solids and Brownness of Each Raw Material

The content of the polyphenolic compound was measured by slightly modifying the Folin-Denis method using a phenomenon in which the phenolic substance reacted with phosphomolybdate to show blue color. That is, 2 mL of Folin-ciocalteu reagent was added to each raw material and 2 mL of these mixed extracts, mixed well, and left to stand at room temperature for 3 minutes. After reacting for exactly 3 minutes, 0.4 mL of saturated Na ₂ CO ₃ solution was added, mixed, distilled water was added to make 4 mL, and the supernatant was measured for absorbance at 725 mm for 1 hour at room temperature. At this time, the total polyphenol compound was determined from the standard curve prepared using tannic acid. This standard curve is obtained by dissolving 1 g of tannic acid in 1 mL of 50% methanol solution and taking final concentrations to 0, 50, 100, 150, 200, 300 and 500 μg / mL solutions. It was prepared by measuring the absorbance at nm.

The total polyphenol compounds contained 0.28% by weight of Ganoderma lucidum, 0.45% by weight of shiitake mushrooms, 1.64% by weight of barberry, 1.77% by weight of barberry, and 0.72% by weight of alder. However, the total polyphenolic compound content of the mixed extracts of the Ganoderma lucidum mushroom, shiitake mushroom, barberry, walnut tree and alder was 1.58%, which was slightly lower than that of barberry and barberry, but in ppm. And 236.4 ppm, much higher than 82 ppm and 88.3 ppm.

Soluble solids were measured with a digital sugar meter (Refractometer PR-101α, Atago Co., Ltd. Japan) and expressed in ° Brix. The brownness of each extract sample was measured OD value at 420 mm absorbance. As shown in [Table 1] below, the absorbance values at 420 nm were 0.478 in hawthorn and hawthorn fruit, respectively, and the highest value was 1.525 in the mixed extract. Therefore, in this experiment, the physiological activity contained in the bark and the fruit and Ganoderma lucidum and shiitake mushrooms will be very useful for the detoxification of alcoholic liver toxicity.

PH, Acidity, Sugar, and Polyphenol Compound Content of Herbal Yogurt Added Ingredients    Kinds   Added amount (% by weight)     pH    Total acidity (%)   Soluble Solids (Brix)    Brownness (420 nm)   Polyphenolic compounds ppm  (weight%) Ganoderma lucidum mushroom 0.250 4.30 0.0032 0.4 0.103 7.0 0.28 Shiitake mushrooms 0.125 5.70 0.0064 0.4 0.042 5.6 0.45 Barn fruit 0.500 4.62 0.0096 0.7 0.478 82.0 1.64 Hut 0.500 5.06 0.0032 0.4 0.478 88.3 1.77 Alder 0.125 4.67 0.0064 0.4 0.098 9.0 0.72 Mixed group ^{1 )} 1.5 4.70 0.0192 1.0 1.525 236.4 1.58

Note 1) The above ganoderma lucidum mushroom, shiitake mushrooms, barberry, barberry, alder extract

3. Each raw material and functional herbal yogurt pH  And acidity measurements

In order to investigate the pH of each raw material and acid production capacity of lactic acid bacteria, 10 mL of the fermentation sample solution was collected by time and measured directly with a pH meter (Methrohm 691 pH Meter, Swiss). The titratable acidity was added to 10 mL of each raw material and fermentation sample solution, and 50 mL of distilled water was added thereto, shaken well, 4-5 drops of 1% phenolphthalein solution was titrated with 0.1 N NaOH, and the consumption was measured and converted into total acidity. The acidity of the extract for each raw material was calculated by the following formula.

Total acidity (%) = (consumption of 0.1 N NaOH (mL) × 0.0064 (citric acid) × factor × 100) / sample (mL)

At this time, the total acidity change of the fermented functional herbal yogurt sample solution was applied to the formula of lactic acid value 0.009.

As a result of measuring the pH and total acidity of each raw material extract, the total acidity was very low in the range of 0.0032 ~ 0.0192%, and the pH was 4.30 ~ 5.70.

The results of measuring the pH of the functional herbal yogurt prepared by the method of 1 are as shown in Table 2 below.

PH change over time during fermentation of functional herbal yogurt Kinds 2 hr 4 hr 6 hr 8 hr 10 hr Ganoderma lucidum mushroom 6.44 5.22 4.45 4.13 3.96 Shiitake mushrooms 6.37 5.28 4.54 4.16 3.93 Barn fruit 6.36 5.05 4.40 4.11 3.96 Hut 6.42 5.19 4.46 4.12 3.97 Alder 6.42 4.96 4.37 4.12 3.97 Mixed group 6.31 4.78 4.29 4.11 3.90 Control 6.34 4.75 4.32 4.05 3.92

As shown in [Table 2], the initial pH of the fermentation broth was about 6.31˜6.44, which was similar in each experimental section. However, due to the concentration of organic acid produced as the lactic acid bacteria were grown, the pH was relatively decreased in all experimental groups. Fermented milk marketed in Korea has a pH range of 3.87 ~ 4.19 and an acidity of 0.97 ~ 1.43%.

The tendency of increasing the acidity during fermented milk production by the addition of single and mixed extracts of herbal and mushroom materials increased steadily in each experimental group until 10 hours after fermentation, similar to the pH change, as shown in Table 3 below. It was. The optimum acidity (10hr) of the fermentation broth was 0.855 ~ 0.954 in each extract addition group, which was the lowest in the mixed extract addition group as 0.972 compared to 0.918 in the no additive control group. These results indicate that acid production is increased by the growth of lactic acid bacteria by various minerals and amino acids contained in the extract, and the proper acidity of normal fermented milk products is in the range of 0.7% to 1.2%.

Changes in Acidity Over Time During Functional Fermentation of Functional Oriental Yogurt Kinds 2 hr 4 hr 6 hr 8 hr 10 hr Ganoderma lucidum mushroom 0.117 0.387 0.522 0.594 0.945 Shiitake mushrooms 0.126 0.360 0.558 0.630 0.900 Barn fruit 0.126 0.378 0.594 0.639 0.927 Hut 0.09 0.378 0.531 0.666 0.855 Alder 0.108 0.450 0.603 0.630 0.954 Mixed group 0.126 0.495 0.639 0.657 0.972 Control 0.117 0.423 0.522 0.630 0.918

4. Functional herbal yogurt Lactobacillus  Measure

Each time, a certain amount of fermented milk was taken and the sample diluted to an appropriate concentration was coated on Lactobacillus ( Mactobacillus lus) MRS agar plates (Difco, Sparks, MD, USA) medium, and the colonies appeared after incubating for 72 hours at 37 ° C. Measurements were made using colony counters. The number of lactic acid bacteria was expressed as CFU (colony forming unit) per mL of fermentation broth.

The number of lactic acid bacteria continued to increase with the fermentation time. The change in the number of lactic acid bacteria did not show a big difference until 4 hours of fermentation, but after 10 hours of fermentation, it showed the lowest value of 1 × 10 ⁸ CFU / mL in the no additive control group and the Ganoderma lucidum extract addition group. Next, alder tree extract addition group 5.4 × 10 ⁸ CFU / mL, shiitake mushroom extract addition group 8 × 10 ⁸ CFU / mL, hollyhock extract addition group 1 × 10 ⁹ CFU / mL, barberry fruit extract addition group 4 It was higher than the control group without the extract in order of × 10 ⁹ CFU / mL [Table 4]. However, as in the pH and acidity change in the fermentation broth, the number of lactic acid bacteria in the mixed extract addition group also appears to be the most to promote fermentation. The number of lactobacillus of fresh liquid and staple fermented milk is more than 10 ⁷ and 10 ⁸ CFU / mL, respectively.

Changes in Lactic Acid Bacteria during Fermentation of Functional Herbal Yogurt (cfu / ml) Kinds 0 hr 2 hr 4 hr 6 hr 8 hr 10 hr Ganoderma lucidum mushroom 1 × 10 ⁷ 1 × 10 ⁷ 4 × 10 ⁷ 3 × 10 ⁷ 6.9 × 10 ⁷ 1 × 10 ⁸ Shiitake mushrooms 1 × 10 ⁷ 2.8 × 10 ⁷ 7.4 × 10 ⁸ 2.9 × 10 ⁸ 3 × 10 ⁸ 8 × 10 ⁸ Barn fruit 1 × 10 ⁷ 5 × 10 ⁶ 8 × 10 ⁷ 5 × 10 ⁸ 1.1 × 10 ⁹ 4 × 10 ⁹ Hut 1 × 10 ⁷ 1 × 10 ⁷ 1 × 10 ⁸ 2 × 10 ⁸ 7 × 10 ⁸ 1 × 10 ⁹ Alder 1 × 10 ⁷ 1.8 × 10 ⁷ 1 × 10 ⁷ 1 × 10 ⁷ 1 × 10 ⁸ 5.4 × 10 ⁸ Mixed group 1 × 10 ⁷ 5.3 × 10 ⁷ 1.1 × 10 ⁸ 1.4 × 10 ⁸ 3 × 10 ⁸ 9 × 10 ⁹ Control 1 × 10 ⁷ 1.6 × 10 ⁷ 3.6 × 10 ⁷ 4 × 10 ⁷ 5 × 10 ⁷ 1 × 10 ⁸

5. Viscosity measurement of functional herbal yogurt

The viscosity of the fermentation broth was measured by fermentation until the end of the fermentation and allowed to cool to 6 ° C. in the state where the curd was completely formed. Viscosity was measured for 1 minute at 60 rpm using a L 3 spindle with a Biscotti Bezick Plus Viscometer (Fungilab S.A., Spain).

As shown in Table 5 below, the viscosity of the fermentation broth of the addition group to which the herbal or mushroom extract was added did not differ from the control without the extract at about 850 to 1000 cps. However, the viscosity of fermented milk in the mixed group was the highest at 1,150 cps. The mixed group showed high viscosity with the same tendency as pH and acidity change in fermentation broth, which can be thought to be due to the relatively high aggregation of casein and whey protein due to increased acid production by promoting the growth of lactic acid bacteria in the mixed group. have.

Viscosity Measurement of Functional Herbal Yogurt Kinds Temperature (℃) cps Ganoderma lucidum mushroom 15 850 Shiitake mushrooms 15 1,000 Barn fruit 15 900 Hut 15 950 Alder 15 950  Mixed group 15 1,150 Control 15 950

6. Color of Functional Herbal Yogurt

For chromaticity, Hunter value L (brightness), a (redness), and b (yellowness) were measured using a colorimeter (Handy colorimeter NR-3000, Nippon enshoku, Japan).

The chromaticity was compared with the control group without the extract because the color showed the best effect in the mixed group. The brightness value (L) of the mixed group was 63.57, which was slightly lower than that of the no addition control 68.66, and the redness value (a) and yellowness value (b) were significantly higher in the fermented milk with the mixed extract, respectively [Table 6]. This can be said that the browning degree (OD 420 nm) in the mixture of raw material extract is 1.525, which is closely related to the much higher than each extract.

Chromaticity value of functional herbal yogurt division L * (brightness) a * (red) b * (yellow) Mixed group 63.57 2.94 11.15 Control 68.66 -2.03 3.86

7. Investigation of the shelf life of functional herbal yogurt

The pH, acidity, and viable cell number were measured in the same manner as when fermentation was completed until 12 days at intervals of 3 days while keeping the fermented samples in the refrigerator.

Unlike other foods, fermented milk is circulated at low temperature for a long time because lactic acid bacteria are alive. In order to determine the quality change during the storage period, the pH, acidity, and viable cell number at 0, 3, 6, 9 and 12 days were stored in the cold storage for 6 ℃ fermented milk after fermentation was completed. [Table 7] to [Table 9] ] Respectively.

As shown in Table 7 below, the pH during the storage period tended to decrease slightly with the storage period, and after 12 days, the pH gradient decreased by 0.04 in the mixed group and the alder extract group, whereas in the no additive control group. The pH was lowered 0.10, indicating the most change.

PH change over time during storage of functional herbal yogurt Kinds 0 day 3 day 6 day 9 day 12 day Ganoderma lucidum mushroom 3.96 3.99 3.92 3.94 3.90 Shiitake mushrooms 3.93 3.99 3.85 3.88 3.86 Barn fruit 3.96 3.95 3.92 3.89 3.87 Hut 3.97 3.97 3.94 3.93 3.90 Alder 3.97 4.01 3.93 3.94 3.93 Mixed group 3.94 3.97 3.92 3.92 3.90 Control 3.92 3.89 3.86 3.83 3.82

      As shown in Table 8 below, the change in total acidity was different from that in the fermentation broth. That is, the pH of the fermentation broth over time tended to be slightly lower, but the total acidity was also slightly lower.

Changes in pH over time during storage of functional herbal yogurt Kinds 0 day 3 day 6 day 9 day 12 day Ganoderma lucidum mushroom 0.945 0.828 0.792 0.765 0.792 Shiitake mushrooms 0.900 0.819 0.846 0.819 0.837 Barn fruit 0.927 0.891 0.801 0.792 0.738 Hut 0.855 0.792 0.792 0.756 0.783 Alder 0.954 0.783 0.783 0.765 0.774 Mixed group 0.972 0.864 0.819 0.846 0.810 Control 0.918 0.927 0.936 0.927 0.900

      On the other hand, the number of lactic acid bacteria changes during storage showed a slight increase in the Ganoderma lucidum and no addition control after 12 days of storage time, while the rest of the experimental group decreased.

Changes in Lactic Acid Bacteria over Time during Storage of Functional Oriental Yogurt Kinds 0 day 3 day 6 day 9 day 12 day Ganoderma lucidum mushroom 1 × 10 ⁸ 1 × 10 ⁹ 1 × 10 ⁸ 5.8 × 10 ⁷ 4 × 10 ⁸ Shiitake mushrooms 8 × 10 ⁸ 1 × 10 ⁸ 1 × 10 ⁷ 7.7 × 10 ⁷ 3 × 10 ⁷ Barn fruit 4 × 10 ⁹ 2.3 × 10 ⁸ 1 × 10 ⁸ 1.1 × 10 ⁸ 1.6 × 10 ⁸ Hut 1 × 10 ⁹ 1 × 10 ⁹ 1.5 × 10 ⁸ 1.1 × 10 ⁸ 6 × 10 ⁸ Alder 5.4 × 10 ⁸ 1.5 × 10 ⁹ 6 × 10 ⁷ 1.2 × 10 ⁸ 2 × 10 ⁷ Mixed group 9 × 10 ⁹ 2 × 10 ⁹ 1 × 10 ⁸ 9 × 10 ⁸ 3 × 10 ⁸ Control 1 × 10 ⁸ 1.9 × 10 ⁸ 6 × 10 ⁶ 6 × 10 ⁷ 2 × 10 ⁸

8. Sensory test

6% (v / v) yogurt (Oriental hebal yogurt, hereinafter referred to as 'OHY') and red grape concentrate (OHY + WGC) and apple concentrate (OHY + AC) were added to the sample. It was added and homogenized and compared with commercially available GUT HD-1 for hangover elimination. At this time, the sensory items were selected for the most preferred sample for each sample by evaluation of overall acceptability, taste, and flavor. Sensation was conducted with 45 researchers (34 males, 11 females) and our research staff consisting of 13 in their 20s, 19 in their 30s, and 11 in their 40s.

As a result of the sensory test, as shown in [Table 10], OHY had the strongest smell because of the strong smell of herbal medicines of hawthorn trees and hawthorn berries used as raw materials, and added red grapes and apple concentrate to taste sweet and fruity flavors. OHY + WGC and OHY + AC showed higher preference than OHY. However, overall taste, aroma, and palatability studies showed the highest preference for commercially available GUT HD-1 products. The reason for this is that yogurt is not yet preferred to smell of herbal medicine.

Sensory test result of fermented milk (Preference number) division OHY OHY + WGC OHY + AC GUT HD-1 Taste One 8 7 29 Flavor One 7 4 33 Overall acceptability One 7 9 28

9. Clinical Trials of Herbal Yogurt

Clinical trials of the herbal yogurt prepared by the above method were carried out over 35 adult men and women over a total of 12 times once a week for 3 months. The 35 participants who participated in the experiment consisted of eight people in their twenties, seventeen in their thirties, sixteen in their 40s, four in their 50s, and the average amount of alcohol consumed was about 1 bottle of soju at one time. Also appeared to drink occasionally. 17 people said that it was effective after drinking hangover and 12 did not feel it at all. Clinical trials were to drink 100 mL of herbal yogurt 1-2 hours before drinking alcohol and 1 bottle of soju in restaurants. As shown in [Table 11], the results of the survey showed that there were no changes in 20 people, slightly increased 14 people, or 1 much. The following is very effective by hangover cancellation effect One, there is considerable effect Six people, there are some effects Seven people who answered that there was no hangover cancellation effect in seven people who answered 19 people in 26 people 79% Indicated.

Clinical Trial Results of Oriental Yogurt Item Investigation contents Gradient gradient (number of people) No change at all Increased slightly Increased a lot  20 14 One Hangover effect (number of people) No A little good very good 7 19 6 One

Example  2. Physiological Functions of Herbal Yogurt with Hangover Relief and Liver Protection

1. Experimental Animal Diet

The experimental group was classified as follows according to the hangover and liver protection functional herbal yogurt added in Example 1 or the like.

-Normal group: A dietary administration normal group containing no functional herbal yogurt and alcohol according to the present invention,

Ethanol group: An experimental group in which only alcohol was administered to a normal group diet,

Ethanol + Gut HD-1 group: Gut group treated with Gut-HD-1 from M Company, which is marketed as a positive control with alcohol administration in a normal diet,

 = OHY group (Ethanol + OHY group): The OHY group treated with the functional herbal yogurt prepared in Example 1 together with the alcohol administration to the normal group diet was carried out by dividing as described in Table 12 below. At this time, the more specific dietary composition of each experimental group is as shown in [Table 13]. At this time, the normal group was drinking the first distilled water, the alcohol group was free to drink all the beverage containing 30% by weight alcohol.

Experimental group and diet Experimental group White rats Dietary Processing Normal 6 General diet Alcohol group 6 Alcohol + General Diet Gut 6 Alcohol + General Diet + GUT HD-1 OHY group 6 Alcohol + General Diet + OHY

Composition percentage of animal experiment diet (% by weight) ingredient Normal Alcohol group Gut OHY group ethanol - + + + Casein 20 20 20 20 α-corn starch 15 15 15 15 Corn oil 10 10 10 10 cellulose 5 5 5 5 AIN-93 Mineral Mixture 4 4 4 4 AIN-93 Vitamin Mixture One One One One L-Methionine 0.3 0.3 0.3 0.3 Sucrose 54.5 54.5 49.5 49.5 Choline Acid Tartrate (dCholine bitartrate) 0.2 0.2 0.2 0.2 Gut HD-1 0 0 5.0 0 OHY 0 0 0 5.0

2. Breeding conditions, sample collection and analysis of test animals

   The experimental animals were purchased for 4 weeks old Sprague-Dawley male rats (Hyochang Science, Korea) and used for this experiment after being adapted to the environment while feeding a commercial solid feed for a week. After one week of adaptation, they are classified into randomized complete block design and placed one by one in a stainless steel cage. Room temperature 22 ± 2 ℃, humidity 50 ± 5%, contrast cycle 12 hours (light cycle: 07: 00 ~ 19:00) was raised in the automatically set animal breeding room. The diet and water containing the dietary components shown in Table 13 above were freely fed, and dietary intake and body weight were measured at regular times every two days during the breeding period.

Animal experiments were reared for four weeks with feeding for each group, followed by fasting for more than 12 hours on the last day of the experiment, and dissected by light anesthesia with ether. After opening, blood was collected from the abdominal aorta, blood was collected, left at room temperature for about 30 minutes, and serum obtained by centrifugation at 3,000 rpm for 20 minutes was used for analysis. After blood collection, each tissue was extracted, washed with cold 0.9% saline solution, dried with filter paper, and weighed.

The results obtained from the experiments were all expressed as mean and standard error (mean ± SE) by one-way ANOVA test, and the significance test between each experimental group was performed by Duncan's multiple range test. Analyzed.

3. weight change

In order to examine the effects of yogurt prepared by adding herbal and mushroom extracts on the effects of alcohol intake on alcoholic liver toxicity, 5.0% (v / The weight change of the rats administered with the semi-synthetic diet added at the w) level for 4 weeks is shown in FIG. 1. Normal weight gained steadily over time with dietary administration. However, in the control group compared to the normal group, the body weight was decreased by the statistically significant difference from the 2nd week of diet administration. However, OHY and Gut HD-1 administration, together with alcohol administration, did not show a statistically significant difference in weight loss compared to the alcohol alone group, but it was shown to offset the weight loss inhibition by alcohol to some extent.

4. Drink Intake

Alcoholic beverages containing 30% alcohol induced alcoholic hepatotoxicity during free trial. As shown in FIG. 2, the beverage intake was higher than that of the other alcohol-containing beverage intake. However, there was no significant difference between the experimental groups who consumed alcohol and drink at the same time, but statistically significant difference from the normal group appeared from the second week.

5. Liver weight

The weight (g) and relative weight (%) of soy sauce are shown in Table 14 below. The increase in liver weight in the alcohol-only group compared to the normal group is a typical result of the alcoholic liver toxicity model, and the previous results indicate that the alcohol toxicity causes cirrhosis, resulting in increased cell volume due to the accumulation of fat, protein and water in the liver cells. Matched. However, this increase in liver tissue weight seen in the alcohol-administered group was significantly reduced by Gut HD-1 or OHY combined administration. This means that the liver weight increased by the administration of alcohol is reduced by simultaneous administration of fermented milk, thereby reducing the amount of neutral lipid concentration which greatly affects the weight of the liver, and at the same time, it can be expected that fatty liver has also been improved.

Liver tissue weight Experimental group Liver weight Gross weight (g) Relative weight (%) Normal 13.27 ± 1.16 ^ab 3.69 Alcohol group 14.56 ± 1.17 ^a 4.36 Gut 11.76 ± 0.73 ^b 3.50 OHY group 12.35 ± 0.26 ^b 3.72

6. Serum Neutral Lipid Concentration

   Trigylceride concentrations of serum isolated after 12 hours of fasting blood on the final day of the experiment were analyzed by Neodine Medical Research Institute, a medical institution. Serum triglyceride concentrations were significantly increased in the alcohol group compared to the normal group as shown in FIG. 3, and this increase by alcohol was significantly reduced to the normal group level by administration of Gut HD-1 or OHY. It was.

7. ADH  And ALDH  Active measurement

Alcohol dehydrogenase (ADH) activity was measured by measuring the rate of production of nicotinamide-adenine dinucleotide + hydrogen (hereinafter referred to as 'NADH') at absorbance of 340 nm. Compared by activity. In other words, the composition of the reaction vessel was adjusted to 1.4 mL of distilled water, 0.75 mL of 1.0 M tris-HCl buffer (pH 8.8), 20 mM NAD ⁺ 0.3 mL, ethanol 0.3 mL, and 0.25 mL of enzyme source to a total of 3 mL, at 30 ° C. After pretreatment for 5 minutes, the change in absorbance at 340 nm was measured for 5 minutes. In this case, the relative activity (%) was expressed as a control without adding an enzyme source. Determination of acetaldehyde dehydrogenase (ALDH) activity is an enzyme that produces acetic acid in acetaldehyde in the liver. Was used. The composition of the reactor was distilled water 2.1 mL, 1.0 M tris-HCl buffer (pH 8.0) 0.3 mL, 20 mM NAD ⁺ 0.1 mL, acetaldehyde 0.1 mL, 3.0 M KCl 0.1 mL, 0.33 M 2-mercaptoethanol (mercaptoethanol ) 0.1 mL of the mixed solution and 0.1 mL of the enzyme source were put in a cuvette, adjusted to a total of 3 mL, and pre-reacted at 30 ° C. for 5 minutes, and then the change in absorbance was measured at 340 nm for 5 minutes. In this case, the relative activity (%) was expressed as a control without adding an enzyme source.

Most of the alcohol absorbed into the body is absorbed by the stomach and small intestine, moved to the liver and then metabolized and broken down. Chronic alcohol intake can cause nutrient absorption and impair liver function. Alcohol metabolism in the liver is first converted to acetaldehyde by alcohol with ADH and cytochrom P-450. This increases acetaldehyde. Chronic alcohol administration in experimental animals increases ADH activity and decreases ALDH activity, suggesting the possibility of acetaldehyde accumulation in biological tissues by prolonged ingestion of alcohol. In this experiment, however, as shown in FIG. 4, the ADH activity was not significantly different between the experimental groups, but the ADH activity was increased by alcohol administration compared to the normal group, and by the administration of GUT HD-1 and OHY It was further shown that the enzyme activity was promoted. These results showed a high correlation with blood alcohol concentration. On the other hand, ALDH activity, which inhibits the accumulation of acetaldehyde, is higher in the Gut HD-1 and OHY administration groups than the alcohol administration group, which is thought to act to inhibit the accumulation of acetaldehyde concentration in the blood. Compared to the normal group, the ADH activity in the liver tissue was significantly increased in the alcohol group because of the adaptability of the liver tissue to activate alcohol metabolism by alcohol consumption. After 1 hour of alcohol intake, ADH activity increased from 36% to 42% in the barberry fruit extract group, compared with 28% in the control group not ingesting the barberry fruit extract. (Kim MH, Chung UT, Lee JH, Park YS, Shin MK, Kim HS, Kim DH, Lee HY. 2000. Hepatic detoxification activity and reduction of serum alcohol concentration of Hovenia dulcis Thunb from Korea and China.Korean H Medicinal Crop Sci 8, 225-233). Acetaldehyde, a major cause of hangovers, is a metabolite produced by alcohol degradation by the enzyme action of ADH, and if the fruit extracts of the hawthorn simply activate the enzymes, the alcohol in the blood may be rapidly reduced, but it remains in the liver or blood. Acetaldehyde may accumulate and cause severe hangover symptoms. ADH and ALDH enzyme activity was 28.8% and 29.8% in the alcohol alone group, and the activity of both enzymes was balanced in vivo to decompose alcohol and its metabolite acetaldehyde. In the same experiment, ADH and ALDH enzyme activity was higher than that of the control group by ingesting the fruit extract of the bark, and in particular, ALDH activity was activated about 10% more than ADH activity, which rapidly decomposed acetaldehyde into acetic acid, which significantly affected hangover resolution. Has been reported. In addition, Ahn et al. Conducted a single alcohol toxicity test on larvae, alders, and mixed extracts that were effective in relieving hangovers. As a result, ADH enzyme activity in liver tissues increased 28% in the control group without the extract. The enzyme activity increased up to 60% in the group which was inoculated with the tree bark extract, but was increased by 60% in the group that had the tree bark + duck (An SW, Kim YG, Kim MH, Lee BI, Lee SH, Kwon HI, Hwang B, Lee HY. 1999. Comparison of hepatic detoxification activity and reducing serum alcohol concentration of Hovenia dulsis Thung and Alnus japonica Steud.Korean J Medicinal Crop Sci 7, 263-268). These results showed that the final alcohol concentration in the blood after 4 hours of alcohol consumption was 32.2% in the larvae extract group and 13.2% in the alder tree extract group compared to the control group of the alcohol alone intake, but decreased by 42.8% in the two mixture extract groups. In the experiments with 20 adult men, the blood alcohol concentration was 0.182% after 15 minutes of alcohol intake, which was similar between groups, but the time taken to drop below 0.004% of the blood alcohol concentration in the normal group of no alcohol intake was extracted from the alder tree. 6 hours, 5 hours of barberry extract, and 4 hours of barberry + duck tree extracts, which are also used to reduce blood alcohol levels after drinking and to lower acetaldehyde levels related to hangovers. Extracts of alder and alder have been shown to promote various enzyme activities, and it is recommended that the development of functional foods using these mixtures will be effective. In the present experiment, the concentrations of alcohol and acetaldehyde in blood were significantly decreased by the increase of ADH and ALDH enzyme activity in the OHY group containing herbal and yogurt extracts compared to the alcohol alone control group. Showed agreement with

8. Improve liver function

ALT, AST, and LDH activity, which is used as an indicator of hepatic function in serum, were analyzed by Neodine Medical Research Institute, a medical institution. Long-term intake of alcohol impairs liver function and is known to cause fatty liver, cirrhosis, liver cancer, etc. The liver function was verified by measuring ALT, AST and LDH activity, clinical indicators of liver damage. Serum ALT and AST activity is a clinical indicator of liver damage, showing high activity as transaminase is released into the blood as necrosis of liver cells and liver tissue destruction due to liver damage progress. Alcohol is mainly metabolized in the liver, and people who consume alcohol frequently increase the destruction of hepatocytes, thereby increasing blood ALT and AST enzyme activity. In this experiment, the ALT activity showed a statistically significant increase in the alcohol group as compared to the normal group. Increasing ALT activity in the blood by alcohol administration showed a similar reduction in the GUT HD-1 and OHY administration groups, and this reduction was markedly reduced to normal levels. Blood AST enzyme activity was 20% higher in alcohol group than normal group, although there was no statistical significance between the experimental groups. This increase was decreased by 24.2% and 16.5% in GUT HD-1 and OHY groups, respectively. LDH activity showed a tendency to increase in all alcohol administration groups compared with normal group without statistical difference. ALT activity decreased by 55.5%, 71.3% and 28.2%, respectively, and AST activity was 65.2%, 75.1%, and 51.2 for D-galactosamine-induced hepatotoxicity. It has been reported to have a hepatotoxic protective effect on mushrooms (Lee EW, He P, Kawagishi H, Sugiyama K. 2000. Suppression of D-galactosamine-induced liver injury by mushroom in rats.Biosci Biotechnol Biochem 64, 2001- 2002). Recently, serum AST activity in BCG-induced immune liver injury mice was significantly reduced by polysaccharide administration of Ganoderma lucidum. After 4 and 8 weeks of fermented milk in healthy adults, serum ALT activity was decreased by 13.6% and 8.9%, respectively (Kawase M, Hashimoto H, Hosoda M, Morita H, Hosono A. 2000. Effect of administration of fermented milk containing whey protein concentrate to rats and healthy men on serum lipid and blood pressure.J Dairy Sci 83, 255-263). Fermented milk containing these mushroom extracts may promote liver function. Experiments to improve liver function deterioration due to diabetic complications showed that streptozotocin-induced type 1 diabetes and genetic type 2 diabetes were induced. Results were shown to reduce serum AST and ALT activity in Zucker obese mice (Jeon BS, Park JW, Shin GG, Kim BK, Kim HK, Cho YS, Cha JY. 2004. Effect of fermented mushroom milk on hyperlipidemia and hepatic injury in streptozotocin-induced diabetic and Zucker fatty rats.Food Sci Biotechnol 13, 576-580). Serum lactate dehydrogenase (LDH) activity, a marker of liver function, was also decreased by 91.1%, 93.2%, and 69.4% in D-galactosamine-induced hepatic injury rats, respectively. Therefore, in the fermented milk prepared by adding these extracts, as well as shiitake and ganoderma lucidum used in this experiment, it was shown that the decreased activity of enzymes that increased liver function due to alcohol intake was effective in activating liver function.

9. Determination of Blood Alcohol and Acetaldehyde Levels

Alcohol and acetaldehyde concentrations in serum were measured using a commercial R-Biopharm UV-Test Kit (Darmstadt, Germany). In other words, the serum alcohol concentration was measured by adding 0.1 mL of serum to a 3 mL mixture of NAD and phosphate buffer (pH 9.0), reacting for about 3 minutes, measuring absorbance at 340 nm, and adding 0.05 mL of ADH to the mixture for 10 minutes at 20 ° C. Absorbance was observed to change the absorbance according to the concentration of NADH produced at 340 nm. At this time, the alcohol concentration in blood (%) was calculated using a standard solution. Serum acetaldehyde concentration was measured by adding 0.2 mL of serum to a 3 mL mixture of NAD and phosphate buffer (pH 9.0), reacting for about 3 minutes, measuring absorbance at 340 nm, and adding 0.05 mL of ALDH to the mixture at 20 to 25 ° C. After reacting for 5 minutes, the change in absorbance according to the concentration of NADH produced at absorbance 340 nm was observed.

Alcohol and its primary metabolite, acetaldehyde, are known to be a major cause of hangovers after drinking alcohol. In this experiment, no blood alcohol concentration was detected in the normal group without alcohol. As shown in FIG. 6, blood alcohol concentrations were detected in all alcohol-administered groups, but significantly decreased to 0.05% GUT HD-1 group or 0.03% to OHY-treated group compared to 0.24% of alcohol-only group. It can be seen that the development of the hangover-resolved fermented milk has been suggested because it has a stronger alcohol decomposition promoting effect in the administration of the OHY product than the commercially available hangover-resolved fermented milk. After fasting 300 g of rats for 1 day, alcohol was diluted in distilled water at a concentration of 8 mg / ml and orally administered at a concentration of 4 g / kg. After 1 hour, the blood alcohol concentration increased to 0.171%. A significant amount of alcohol remained in the blood at 0.152% after time (An SW, Kim YG, Kim MH, Lee BI, Lee SH, Kwon HI, Hwang B, Lee HY. 1999. Comparison of hepatic detoxification activity and reducing serum alcohol concentration of Hovenia dulsis Thung and Alnus japonica Steud. Korean J Medicinal Crop Sci 7, 263-268). In this experiment, alcohol was still present in the blood because of continuous consumption of alcohol with ingestion of beverages, as well as ganoderma lucidum, barberry, alder, and ganoderma lucidum and shiitake with liver protection. Significantly lowered blood alcohol levels by the administration of functional herbal yogurt made with the addition of mushroom extracts has been shown to be superior to any hangover-relieving beverages containing only extracts of these herbal materials with alcohol-degrading effects. (Barak AJ, Beckenhauer HC, Badakhsh S, Tuma DJ. 1997.The effect of betaine in reversing alcoholic steatosis.Alcohol Clin Exp Res 21, 1100-1102, Yang DS, Hong SG, Choi SM, Kim BN, Sung HJ, Yoon YS. 2004. Effect of an oriental herbal composition, Jang-Baek-Union (JBU), on alcohol-induced hangover and CCl4-induced liver injury in rats J Korean Soc Food Sci Nutr 33, 78-82).

In general, alcoholic beverages made by adding fruit juice or spices to the alcoholic beverages, which is a favorite food, have different odors even though they consume the same amount.Also, when drinking alcoholic beverages containing various natural products May appear differently. This is because the mixing of alcohol with other substances affects the alcohol metabolism, in which case the additives may promote or delay the alcohol metabolism. However, the systematic and scientific verification of the efficacy of alcoholic beverages made by adding various functional substances to the human body is still insufficient.

Acetaldehyde is a potent toxic substance that acts as a major mediator of liver damage through condensation with active amines in vivo and affects hangover. It is metabolized by ALDH to form acetic acid and then hydrolyzed with carbon dioxide and water to completely decompose it. You will go through the process. Toxicity of acetaldehyde includes cirrhosis and impaired heart and brain function due to mitochondrial impairment. In the acute toxicity alcohol test, acetaldehyde concentration was about 1,000 mg / L in the ethanol control group, and it was reported that there was a 75 ~ 86% reduction effect in the oriental medicine group consisting of 10 herbal medicines including Goji (Yang DS, Hong SG, Choi SM, Kim BN, Sung HJ, Yoon YS. 2004. Effect of an oriental herbal composition, Jang-Baek-Union (JBU), on alcohol-induced hangover and CCl4-induced liver injury in rats.J Korean Soc Food Sci Nutr 33, 78-82). In the present experiment, the concentration of acetaldehyde in blood was significantly increased in the alcohol-administered group compared with the normal group, which was shown to increase the intermediate acetaldehyde by alcohol intake, and this increase by alcohol was observed in Gut HD-1 and OHY. The administration group showed a tendency to decrease slightly. These results have been shown to have a high correlation with the low alcohol levels in the blood resulting from the consumption of fermented milk, and the rapid alcohol degradation after drinking also lowers the acetaldehyde concentration, which is known to be the cause of hangover. Suggests.

10. Preparation of fraction of liver tissue and measurement of lipid peroxide

Homogenate fraction preparation from liver tissue was prepared by grinding a portion of the liver and adding 4 times the amount of homogenate solution containing 250 mM sucrose. The homogenate solution was centrifuged at 10,000 x g for 20 minutes in a high-speed centrifuge (VS-24SMTi, Vision Scientific Co., Ltd.) to obtain a supernatant containing microsomes to provide for lipid peroxide measurements. Homogenate And Protein concentration of the microsomal fraction was measured using bovine serum albumin as a standard according to the Lowry method. The lipid peroxide content of the fractioned homogenate and microsomal biofilm was quantified as follows. That is, 2 ml of thiobarbituric acid (TBA) reagent was added to 1 ml of the fraction solution containing 1 mg of protein, mixed well, heated for 30 minutes in a water bath, and allowed to cool at room temperature to 3,000 rpm, 10 The supernatant centrifuged for a minute was measured for absorbance at 535 nm. Lipid peroxide content was expressed as n mol / g of marondialdehyde (malondialdehyde).

Lipid peroxidation is recognized as the most important mechanism of liver damage caused by various toxic compounds and drugs. This mechanism is well known to be due to an increase in oxidative stress caused by increased intracellular free radical production and a decrease in antioxidant defenses. Liver tissue plays a major role in reducing toxicity by decomposing all the toxic compounds absorbed into the body and exogenous toxic compounds, including alcohols and drugs, so there are many opportunities for exposure to various toxic substances in the process. Known as an accumulation site, this can lead to tissue damage by lowering the antioxidant system and providing a good environment for producing lipid peroxides. The TBARS content, known as an indicator of the peroxide production of biofilm lipids in the animal body, was statistically increased in the homogenate fraction of liver tissues compared to the normal group. Gut HD-1 and OHY administration significantly reduced (FIG. 7). In the microsomal fraction of liver tissue, the TBARS content was the highest in the alcohol group but decreased to the normal group level in the Gut HD-1 group, while in the OHY group, there was a slight difference in activity between the two fermented milks. To date, such natural antioxidants include tocopherols, carotenoids, phenolic compounds (flavonoids, catechins, tannins) and sulfur-containing compounds. Flavonoids such as (+)-ampelopsin, laricetin, myricetin, and (+)-gallocatechin and hovenitin were also used in the barn tree used in this experiment. ) Isolation of flavonols such as Ⅰ, Ⅱ, and Ⅲ has been identified, and physiological activity of these components has been shown to cause D-galactosamine, lipopolysaccharide, and carbon tetrachloride-induced liver damage. Toxicity inhibitors and antioxidants have been reported. Alder trees also contain several flavonoids and tannins, which have been known to act as antioxidants. The total polyphenolic compounds of each raw material used in OHY also contain 0.28% by weight of Ganoderma lucidum mushroom, 0.45% by weight of shiitake mushroom, 1.64% by weight of hawthorn fruit, 1.77% by weight of hawthorn, and 0.72% by weight of alder. Was fed.

Functional herbal yogurt according to the present invention is obtained by mixing the mushroom material ganoderma lucidum mushroom and shiitake mushrooms with excellent protection of liver function in addition to the herbal material material hawthorn, barberry, alder, which has a hangover effect by lowering blood alcohol concentration By using water-soluble extract, it lowers blood alcohol level and acetaldehyde level due to alcohol intake, promotes alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activity of alcohol and acetaldehyde degrading enzyme in liver tissue, liver function It can be seen that the clinical indicators of serum AST and ALT activity, hepatic tissue neutral lipid concentration decrease, oxidative stress inhibitory effect by decreased lipid peroxidation concentration, alcoholic hepatotoxicity and fat degeneration effect by histological observation.

In addition, the functional herbal yogurt according to the present invention is more effective against alcoholic liver toxicity by adding mushrooms in addition to medicinal herbs, hawthorn fruit, alder, which are herbal materials, as well as hepatotoxicity due to alcohol intake, especially in Korean men It will also contribute to improving or preventing liver disease. In particular, the extract obtained by the appropriate mixing ratio of herbal and mushroom materials significantly increases the number of lactic acid bacteria in fermented milk during fermentation process, and by ingesting them, the greatest advantages that lactic acid bacteria can have, intestinal environment improvement, immunity enhancement, allergy reduction, prevention of tooth decay, In addition to various effects, such as cholesterol reduction, it can be expected to have an additional effect of promoting metabolism due to the intake of the vitamin B group produced during fermentation.

Claims (5)

Ⅰ) Thinly cut dried bark and alder into 94.0 ~ 99.8% by weight of water, add 0.1 ~ 2.0% by weight and 0.1 ~ 2.0% by weight of barberry, and boil for 30 minutes to 2 hours at high temperature of 98 ~ 121 ℃. Obtaining an extract of the components contained in the tree and the fruit (medicinal herb component extraction step); Ii) chopped dried shiitake mushrooms and dried ganoderma lucidum in 99.0 ~ 99.5% by weight of water extracted from boiled barn tree, barberry, alder in step iii) and cut the size of horizontal and vertical 0.5 ~ 1 cm Putting 0.05 ~ 1.0% by weight shiitake mushrooms under the same extraction conditions as in step iii) to extract the ingredients contained in Ganoderma lucidum and shiitake mushrooms (mushroom ingredient extraction step); Iii) preparing a lactic acid bacterium fermentation broth by mixing 7-15% by weight of skim milk powder (based on solids) with 85 to 97% by weight of the extract extracted from the herbal and mushroom components prepared through steps iii) and ii) step); Iii) pre-culturing the mixed lactic acid strain in the lactic acid bacteria culture medium which has been sterilized separately at 80 ° C. to 100 ° C. (mixed lactic acid strain culture step); Iii) 1 ~ 3% by weight of the starter (starter) pre-cultured the lactic acid strain mixed in step iii) to 97 ~ 99% by weight of the lactic acid bacteria fermentation broth prepared in step iii) to about 8 ~ at 37 ± 1 ℃ Incubation for 12 hours (lactic acid bacteria fermentation step); Method of producing a functional herbal yogurt, characterized in that is produced through. The method of claim 1, Method of producing a functional herbal yogurt, characterized in that the mixed amount of the bark and alder in the step iii) is 75 to 85% by weight versus 15 to 25% by weight. The method according to any one of claims 1 and 2, Extraction in step iii) is a method of producing a functional herbal yogurt, characterized in that the extraction once or twice repeated filtration using gauze. The method according to any one of claims 1 and 2, The mixed amount of shiitake mushrooms and dried ganoderma lucidum in step ii) is 40 to 60% by weight versus 40 to 60% by weight Method for producing a functional herbal yogurt, characterized in that preferred. The method according to any one of claims 1 and 2, The mixed lactic acid strain in step iii) is Lactobacillus acidophilus , Lactobacillus thermophilus , Bifidobacterium longgum ( Bifidobacterium) loggum ) method of producing a functional herbal yogurt, characterized in that.

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