KR20080109116A - Beverage for relieving hangover and manufacturing method thereof - Google Patents

Abstract

The beverage for hangover relief is provided to inhibit the activity of alcohol dehydrogenase as primary alcohol-degrading enzyme, and suppress the production of acetaldehyde as the oxidizing substance of alcohol, thereby protecting liver and inhibiting hangover phenomenon. The beverage for hangover relief comprises the extracts of Alnus japonica, Citrus unshiu, Glycyrrhiza uralensis, Hovenia dulcis and Pueraria thunbergiana. The herb medicine extracts are prepared by extracting the lath(Alnus japonica), the citrus peel(Citrus unshiu), the glycyrrhizae radix(Glycyrrhiza uralensis), the earth bottle(Hovenia dulcis) and Puerariae Flos(Pueraria thunbergiana) in ethyl alcohol.

Description

Hangover drink and its manufacturing method {Beverage for relieving hangover and manufacturing method

Figure 1 is a photograph showing the antimicrobial activity test results of the Earth disease extract,

Figure 2 is a photograph showing the antimicrobial activity test results of gallium extract,

Figure 3 is a photograph showing the antimicrobial activity test results of wood vinegar.

The present invention relates to a hangover drink and a method for producing the same, and more particularly to a hangover drink and a method for producing the hangover extract as a raw material.

Along with the economic growth and the extension of life expectancy, the perception of modern people's quality of life due to disease and aging society is changing, and there is a great interest in natural products with physiological activities such as antioxidant, antibacterial, anticancer and immune enhancing activities. (Das et al., 2002: Lee and Lee; Park and Kim, 1992).

The palatability of beverages is also changing, and the consumption of cola and cider, which is simply refreshing, is decreasing, and the sales volume of products mainly made from natural products is increasing every year (Park et al., 1998). In recent years, as health concerns are heightened, health drinks containing functionalities while developing pharmacological components, which are inherent components of drugs for treating diseases, have been developed. As a result, natural ingredients such as vegetables and herbs are mainly used (Seo and Kim, 2001). In this context, the development of hangover drinks is also active.

Alcohol is a product that has been handed down to mankind since the beginning, and drinking is a social behavior that is relatively persistent among various social habits (Health, 1990). Alcohol acts on the central nervous system of the brain to make you feel better and to forget the pain, so in ancient times alcohol was used as the primary excipient for all drugs (Marshall and Fritz, 1989).

Alcohol is absorbed into the body at the time of ingestion and is distributed evenly throughout the body, and most of the alcohol in the liver, alcohol dehydrogenase (ADH), acetaldehyde dehydrogenase (ALLDH), endoplasmic reticulum (microsomal ethanol) It is metabolized by certain enzymes such as oxidizing system (MEOS), catalase and peroxidase, and less than 10% of the absorbed alcohol is excreted through the kidneys or lungs without being metabolized.

However, symptoms such as nausea, vomiting, dizziness, thirst, lethargy, headache, and muscle pain, such as acute alcoholic hangover symptoms caused by high intake of alcohol, cause socioeconomic damage due to decreased work efficiency (Swift and Davidson, 1998), Chronic alcohol intake causes disorders such as pancreatitis, myocardial infarction, neuropathy, and tuberculosis, and can lead to fatal damage to the structure and function of liver tissue (Lieber and Leo, 1992; Liever et al., 1986). Therefore, the effect of alcohol on the liver is very large compared to other organs in the mine (Goodman and Gilman, 1975; Lieber and DeCarli, 1968; Lieber et al., 1986).

Currently, the liver dysfunction caused by excessive drinking or frequent drinking, which is a characteristic of Korean drinking culture, is increasing day by day. Therefore, prevention and healing of various liver diseases appearing as one of the important research tasks, Much attention is paid to drugs or beverages that can eliminate hangovers.

In addition, in connection with the elimination of hangovers, various dietary supplements containing ingredients extracted from natural foods or herbal medicines have been developed (Kim Jung-han et al., 38 (6): 549-553, 1995). As part of this research, a composition containing choline acids, which have an effect of edema, hepatic detoxification, hepatic blood flow improvement, promoting fat absorption, and excretion of waste products through fine bile ducts, is commercially available.

Examples of such choline acid include ursodeoxycholic acid, which is known to be effective in improving liver function of liver disease, systemic malaise due to liver dysfunction, indigestion, anorexia, physical fatigue, and the like; Tauroursodeoxycholic acid, which is commercially available as a gallstone dissolving agent and is effective in treating fatty liver, which has an effect of improving liver function and reducing liver fat mass when orally administered; Chenodeoxycholic acid sold as a gallstone dissolving agent; And dehydrocholic acid.

On the other hand, as a herbal medicine or folk remedies, various types of medicines have been developed, which are mainly composed of ingredients such as ginseng and thorny ginseng, which are known to have a beneficial effect on hangover. Among them, a number of nourishing tonic drinks are mainly marketed as ginseng, and in addition, various components such as honey and vitamins are also formulated in an appropriate amount and marketed as nourishing tonics. Ginseng, Brykhman et al., Lloydia, 32 (1): 46-51, 1969, Ocimum sanctum Linn, tinosporama labarica (Sen, P., Maiti, PC and Ray, A. Indian J. Exp. Biol., 30: 592-596, 1992), and herbal substances such as melatonin and other biological substances have been reported to exhibit hangover remedy. Among the symptoms, only some symptoms were effective, or the hangover effect was found to be insignificant.

Recently, a product containing a plant extract mainly containing bean sprouts and a product containing a germ and soybean extract have been introduced. However, these also pointed out many problems that still need to be solved in terms of the stability of the formulation.

Therefore, in order to solve this problem in the field, there is a demand for the development of a new hangover removal composition having an effect on the hangover while ensuring stability.

The present invention has been made to solve the problems of the prior art as described above, by using a herbal material, the invention inhibits the activity of alcohol dehydrogenase which is the primary enzyme metabolizing alcohol in the liver of the acetaldehyde which is an oxide of alcohol It is an object of the present invention to provide a hangover-relieving beverage and a method for producing the same, which can suppress the production and thereby suppress the liver protection effect or the hangover phenomenon.

In order to achieve the above object, the present invention is an active ingredient extracted from the alder tree (Alnus japonica), citrus (Citrus unshiu), licorice (Glycyrrhiza uralensis), Earth disease (Hovenia dulcis) and brown (Pueraria thunbergiana) as an active ingredient It provides a drink composition for hangover relief characterized in that.

Hereinafter, the present invention will be described in detail.

Licorice (甘草, Glycyrrhiza uralensis) extract contained in the hangover drink of the present invention is a useful plant that is well known and widely used as a herbal medicine, triterpene-based saponins, glycyrrhizin, Glycyrrhizin, Liquirition (Liquirition) ), Asparagin, Glutamic Acid, Glucose, etc. 2-glucuronic acid glycoside of glycyrrhizic acid is a sweetener component of licorice and is known to help detoxification in liver because it has various detoxification effects for drug poisoning, plant poisoning, poisoning of body metabolism as well as bactericidal disinfection. have.

The extract of Pueraria thunbergiana included in the hangover drink of the present invention includes flavonoids, essential oils and benzoic acid, benzoic acid, propionic acid, coumaric acid, and the like, such as rovinin, genistein, isoflavone, dydzein, and quercetin. It is. Rovinin has been known to be mainly related to detoxification due to diuretic effect and reduction of residual nitrogen content in blood, and Dyzedine is known to act on various heart failures such as migraine, hypertension and angina. .

Alnus japonica included in the hangover drink of the present invention belongs to the birch family (Betulaceae) and is also known as Yurimok (楡 理 木) in Chinese, and is called Chajo (茶 條). The young leaves and bark of alder trees are called red lambs. Spring and autumn are collected and dried in the sun. Its main ingredients include large amounts of triterpenoids such as lupenone, beta-amylin, glutenol, taraxerol, and betulinic acid. Β-sitosterol, fatty alcohols, pyrocatechol-based tannins are contained. Its effects are known to soothe burns and traumatic bleeding, and to clear and strengthen.

Tangerine blood (Citrus unshiu) included in the hangover drink of the present invention is collected by drying only the peel of tangerine as commonly seen by us and is also called dermis. The tangerine is effective in releasing clumps, strengthening the function of the spleen, removing moisture, and at the same time scavenging the walls. In addition, the tangerine contains aspirin, askulete, tannin, etc., is effective in relieving, asthma, antitussive, expectoration, anti-inflammatory pain, urine and uric acid to help the discharge of alcohol in the body quickly To be done.

Earth disease (Hovenia dulcis) included in the hangover drink of the present invention is also known as a hut tree, contains malic acid, glucose, calcium, habenic acid, furclinine, hobenin, hobenoside, and is excellent in alcohol detoxification Known. In particular, the earth fruit, the fruit of the earth disease, fructose, glucose, catalase, peroxidase, potassium, calcium, iron, triterpenoids, hobeic acid, rutin, saponin, apelopsin, etc. It is known to be effective in relieving and relieving liver toxicity.

In particular, the present invention is characterized by containing all the extract of the above-described raw materials. The present invention will be described in detail by the following examples, but these examples are only for illustrating the present invention, and the scope of the present invention is not limited to these examples.

Production Example : Preparation of Extract

600g of dry herbs of licorice, brownish, alder, tangerine, and earth diseases are added with 6,000ml of 40% ethyl alcohol A (95% alcohol: distilled water = 2: 3), which is 10 times higher. Extraction cycle, rice bran company) was extracted for 120 minutes. The extracted material was filtered through a nonwoven fabric, and then concentrated under reduced pressure to 1,000 ml using a rotary vacuum evaporator (Eyela N-1000, Japan) and centrifuged (10,000 × g, 40 min, 4 ° C.). The supernatant obtained at this time was divided by 1/10 and lyophilized and stored at 4 ° C. In addition, the measurement of the extraction yield for the herbal medicines used in the experiment is shown in Table 1 as a percentage of the total soluble solids content of the extract for 600g herbal medicines.

Table 1. Extraction yield of each test plant with 40% ethyl alcohol

Scientific name Extraction yield (%) Alder 4.9 Tangerine 21.6 licorice 13.8 Earth disease 22.0 Gala 20.6

Example  1 and Comparative example  1 to 3: Preparation and purchase of extract

The hangover beverage composition prepared in this experiment was extracted using the same extraction solvent as the method of extracting the herbal medicine by combining the herbal medicine and wood vinegar solution shown in Table 1 as the main ingredients, and prepared by adding a small amount of honey, oligosaccharide and citric acid. (Haejuga). In addition, for the comparison between the hangover drink composition of the present invention and the hangover drink sold in the past, the hangover drink sold by A, B, C companies were purchased in the market and used in the experiment.

Test Example 1 : Determination of Alcohol Dehydrogenase Activity

1. Experimental method

1) Determination of yeast ADH activity

The activity of yeast ADH (Sigma Co., St. Louis, MO, USA) was modified by the previous method (Jung et al.,) And absorbance of NADH formed at 340 nm using a UV spectrophotometer. Was measured as follows. 40 units / ml ADH, 13 mM NAD ⁺ free acid (Sigma Co., St. Louis, Mo., USA) was used after storage at −20 ° C., tris-base (Sigma Co., St. Louis, MO, USA), 0.05 M Tris-buffer (pH 8.8) was used while stored at 4 ° C. (Kim and Zeikus, 1992)

In a UV cuvette, 0.05M Tris buffer was added to 0.008ml of ADH, 0.04ml of ethanol, and 0.08ml of NAD to make a total volume of 1ml. 50 mg herbal extracts were suspended in 1 ml distilled water and centrifuged in order to see the inhibition or activation of ADH on the extract samples. 10 μl of the supernatant was added to the enzyme reaction solution. Beverages A, B, and C were added at a concentration of 45 μl to measure the enzyme activity. The activity of ADH was calculated for the relative ADH activity (%) indicating the activity of ADH when the extract or hangover beverage was added to the activity of ADH indicated in the control test.

2) Determination of rat liver ADH activity

Sprague-Dawley males weighing about 260 g were used to measure ADH activity in rat liver. After the rats were guillotine, the abdomen was excised to remove the liver. After removal of the nuclear fraction by adding 10 mM Tris buffer (pH 7.4) equal to 10 times the organ weight (0.6 g) of the extracted liver, homogenizing and centrifuging at 700 × g for 10 minutes, the supernatant was collected at 10,000 × g. After centrifugation for 20 minutes, the supernatant was ultracentrifuged at 105,000 × g for 60 minutes. The cytosolic fraction, the supercentrifuged supernatant, was used as the ADH enzyme solution (Kim and park, 1997).

The activity of the prepared rat liver ADH was measured by adding 33 mM sodium phosphate buffer (pH 8.0) containing 1.0 mM NAD ⁺ and 5 mM ethanol to UV cuvettes and adding 0.05 ml of rat hepatocellular fraction to a total volume of 1 ml. It was a test. The activity of rat liver ADH on the extract sample was performed in the same manner as in the assay of yeast ADH activity.

2. Results and Discussion

Table 2. Relative ADH activity (%) of 40% ethyl alcohol extract of medicinal plants obtained

Relative activity (%) East ADH Rat liver ADH Alder 9.8 ± 11.41 47.4 ± 3.5 Tangerine 21.0 ± 11.68 69.3 ± 3.2 licorice -5.6 ± 6.2 54.6 ± 2.3 Earth disease 31.9 ± 14.0 89.7 ± 6.0 Gala 8.1 ± 11.7 38.3 ± 2.4

Table 2 shows the inhibitory activity against yeast ADH in all five herbal extracts. Referring to Table 2, the magnitude of the activity was found to be in the order of licorice ≥ browning ≥ mallard ≥ tangerine ≥ earth disease in the order of inhibitory activity in a similar trend for ADH of rat liver cytosol (cytosol). In particular, browning, alder, and licorice showed a strong activity that inhibited more than 90% of yeast ADH. These results showed that ADH was very similar to previous studies (Lee and Lee, 1999; Moon et al., 2004). Results indicate activity.

Table 3. Relative ADH activity (%) of hangover beverage

Relative activity (%) East ADH Rat liver ADH Do it 13.6 7.7 Company A 19.5 22.6 Company B 72.0 45.9 Company C product 87.4 7.7

Table 3 shows the inhibitory activity against ADH of yeast ADH and rat liver citsol fractions using the hangover and other company's hangover prepared in this study. Referring to Table 3, Haeda showed similar ADH inhibitory activity as that of Company A, whereas Company C exhibited not very high ADH inhibitory activity in vitro.

Extracts and hangover beverages that showed strong inhibitory activity against yeast ADH, in vitro experiments, showed a slight decrease in the cytoplasmic fraction of rat liver, which is more in vivo. The results suggest that (Lee and Lee, 1999) support the idea that the activity test results for purely isolated enzymes can be used to estimate biological activity. In addition, the results of these drugs, which have been used for the treatment of liver disease and the protection of liver in oriental medicine and folk medicine, without clear active ingredients or mechanisms of action, showed inhibitory activity against ADH. It can be estimated that drugs used as therapeutic and protective drugs may also exhibit inhibitory activity of ADH.

Test Example 2 : DPPH Electron Donation ability  Measure

1. Experimental method

Antioxidant activity of each sample was measured by DPPH ((1,1-diphenyl-2-), which loses its color by reacting with an antioxidant using nitrogen accepting hydrogen atoms in an unstable state of hydrazyl. A method using the characteristics of picrylhydrazyl, Sigma Co., St. Louis, MO, USA) was measured by modifying the previous method (Blois, 1958; Lee et al., 2002: Moon et al., 2004). 990 μl of ethanol 1.5 × 10 ⁻⁴ M DPPH was vortexed with 10 μl of extract for each concentration, and the absorbance was measured at 517 nm after 3 minutes, using the solvent used to dissolve the sample instead of the sample. The electron donating ability (EDA,%) of DPPH was calculated by the following equation.

EDA (%) = (B-A) / B × 100

A: absorbance of the sample at 517 nm

B: absorbance of the control at 517 nm

The concentration of EDA (%) is the sample extract necessary for the DPPH concentration of 50% reduction in the time 3 minutes after the sample is added from the concentration and the equation obtained from the correlation between the erasure rate back obtained by the concentration and amount of each sample (RC ₅₀₎ Was calculated. In addition, the antioxidant activity was compared using a fat-soluble antioxidant (+)-α-tocopherol (Vitamin E, Sigma Co., St. Louis, MO, USA) as a comparison.

2. Evaluation Criteria

In general, there are many methods for measuring the antioxidant activity of a specific substance. Among them, the DPPH radical scavenging activity method is relatively simple and can be measured in large quantities. This material is a relatively stable compound with radicals and develops purple in ethanol solution. However, when encountering a substance having antioxidant activity, the antioxidant activity can be easily measured by using the fact that the antioxidant active substance discolors by scavenging radicals of DPPH. Therefore, it has the advantage of being highly associated with the antioxidant activity of the unknown plant extract, and also has the advantage of being highly associated with the antioxidant activity (Song et al., 2000). In other words, the electron donating action is used not only as a measure of inhibiting lipid oxidation by donating electrons to active radicals, but also as a measure of inhibiting disease and aging caused by active radicals in the human body (Choi and Oh, 1985). Harman, 1987)

3. Results and Discussion

Table 4. Electronic donating ability of medicinal plants obtained, 40% ethyl alcohol extract of wood vinegar and hangover beverage

Antioxidant Activity RC ₅₀ (µg / ml * or µl **) Alder 24.7 ± 0.9 Tangerine > 1,500 licorice 1,055.3 ± 21.2 Earth disease 1,126.7 ± 45.1 Gala 460.4 ± 10.5 (+)-α-tocopherol 14.7 ± 1.5 Wood vinegar 75.7 ± 4.0 Company A 9.4 ± 0.5 Company B 36.2 ± 1.9 Company C product 120.4 ± 12.8 Do it 4.1 ± 1.7

Table 4 shows the scavenging activity of DPPH of extract samples of different Chinese herbs, hangover quenching drinks prepared by other companies and the laboratory at various concentrations, and shows the RC ₅₀ calculated from the relationship between absorbance and scavenging rate. Referring to Table 4, the RC ₅₀ of (+)-α-tocopherol used as a control of the present study was 14.7 ± 1.5 μg / ml, which is in good agreement with the previous study (Lee et al., 2002). The results of the study showed effective activity at the concentrations of 460.4 ± 10.5 and 24.7 ± 0.9 ㎍ / mL in browning and alder, especially RC ₅₀ of Alder extract was 24.7 ± 0.9 ㎍ / mL, which was almost the same as that of (+)-α-tocopherol. Similar free radical scavenging effects were observed. These results support the findings of previous studies (Lee et al., 2003) that the alders contained large amounts of 10 diarylheptanoids with significant antioxidant activity. The licorice extract of this study showed a low antioxidant activity of RC _{50 which} is about 2 times higher than 70% methanol extract of previous studies (Lee et al., 2002). As a result, there was a difference in antioxidant activity, that is, hot water extract showed the strongest sulfated activity, and the antioxidant activity of methanol extract was higher than that of ethanol extract (Dong et al., 2004: Kim et al., 1996; Kim et al., 2004), and the trend is expected to show similar results in the global disease.

RC ₅₀ of wood vinegar was 75.7 ± 4.0 μl, which was 10 times lower in antioxidant activity than previous studies (Jeong and Shim, 2002). , The effect of moisture content and maturation period in wood (Alen et al., 1996; Guilled and Ibargoitia, 1999; Guilled and Ibargoitia, 1999; Ku et al., 2002; Maga and Chen, 1985; Toth and Potthast, 1984) It is thought that the wood vinegar used in the manufacture of hangover drink should use wood vinegar with certain quality and ingredients.

DPPH-free radical scavenging activity of hangover drink showed the highest antioxidant activity of RC ₅₀ of Haegagaga prepared in this study, with 4.1 ± 1.7 μl. Indicated. Therefore, the hangover drink prepared in this study is expected to act as a functional drink of excellent natural resources with anti-oxidant activity from the viewpoint of disease and aging control as well as hangover action of other company's hangover drink after drinking.

Test Example 3 : Antibacterial activity measurement

1. Experimental Materials

1) Use strain and medium

For the measurement of the antimicrobial activity of the extracted samples, Bacillus subtilis PM125, Micrococcus luteus KCTC 1056 and Staphyloccus aureus KCTC 1916 and antibiotics were identified as gram-positive bacteria. Methicillin resistant Staphylococcus aureus (MRSA) CCARM 3561, 3115 and 3809, resistant strains to Methicillin were used. Gram-negative bacteria include Escherichia coil D 31, Enterobacter aerogenes KCTC 2190, Klebsiella pneumoniae KCTC 2208, Pseudomonas aeruginosa KCTC 2004 and Mouse Salmonella typhimurium KCTC 1925 was used, and marine bacterium Vibrio parahaemolyticus KCTC 2471 was used.

The growth medium for pre-culture and main culture of Gram-positive and Gram-negative bacteria was tryptic soy broth (TSB, Sigma Co., Darmstadt, Germany), and TSB Yeonhancheon (TSB) was used as a solid medium. soft agar) (TSA, Sigma Co., St. Louis, Mo., USA). In addition, the growth medium for preculture and main culture of marine bacteria used TSB and TSA containing 1% sodium chloride.

2. Experimental method

1) Measurement of growth inhibition of extract

Antimicrobial activity of the extract sample was performed by modifying the disc-agar plate diffusion method of the previous method (Gavidson and Parish 1989; Turner et al., 1998). The fungi cultured up to the mid-logarithmic phase were diluted so that the OD ₅₇₀ value was 0.1 (5 × 10 ⁷ CFU / mL), and the solution was uniformly applied by dropping 0.1 mL of the fungus onto TSB (Tryptic soy broth). Each extract sample was taken at a concentration of 1 mg / ml, dissolved in 50 μl of sterilized deionized water, and completely absorbed into a paper disc sterilized with 75% ethanol (thickness 8 mm diameter, Avantec Toyo, Japan). Put on top. Gram-positive bacteria and Gram-negative bacteria were cultured for 12 hours at 37 ° C, and the diameter of the clear zone formed around the disc was measured (mm). Enteritis Vibrio KCTC 2471, a marine bacterium, was cultured at 30 ° C for 18 hours. After that, the diameter of the growth inhibitory rings (mm) was measured and compared.

2) Minimum inhibitory concentration (MIC) measurement

The measurement of the minimum inhibitory concentration (MIC) of the extract samples showing the antimicrobial activity was determined by the liquid growth inhibition assay (Bullet et al., 1993) using a 96-well plate. same. Each of the bacteria was incubated in a logarithmic growth stage (OD ₆₃₀ = 0.4, 5 × 10 ⁷ CFC / ㎖) in TSB medium at 37 ℃. After mixing the culture medium and the serially diluted sample extract in a 96-well plate (well plate) and incubated for 18 hours at 37 ℃, measuring turbidity at 630nm using ELISA MQX200 (Bio-Teck instrument, USA) Afterwards, the minimum concentration at which bacterial growth was inhibited was defined as the minimum inhibitory concentration for the test bacteria. The minimum inhibitory concentration for marine bacteria was measured in the same manner as for the other strains except for incubation at 25 ° C using TSB medium containing 1% NaCl.

3. Results and Discussion

1) Measurement of growth inhibition of extract

Table 5. Growth inhibition indicated by 40% ethyl alcohol extract of a number of herbal medicines against B. subtilis PM 125, M. luteus KCTC 1056 and S. aureus KCTC 1916, MRSA CCARM 3561, 3115 and 3809 as Gram-positive bacteria

Growth inhibition ring B. Subtilis M. Luteus S. aureus MRSA 3561 MRSA 3115 MRSA 3809 One* 5 10 One 5 10 One 5 10 One 5 10 One 5 10 One 5 10 Alder - - 10 - 10 14 - - - 10 12 15 15 21 24 12 19 20 Tangerine - - - - - - - - - - - - - - - - - - licorice - - 10 - - - - - - - 11 14 - 13 16 - 13 16 Earth disease - - - - - - - - - - - - - 10 12 - - 10 Gala - - - - - - - - - - - - - 12 15 - 12 14

* Concentration of extract of medicinal plants, mg / ml

** not measured

Table 6. As a Gram-negative bacterium, a number of herbal medicines 40% ethyl for E. coli D31, E. aerogenes KCTC 2190, pneumococcal KCTC 2208, Pseudomonas aeruginosa KCTC 2004, and enteritis vibrio and marine bacteria enteritis vibrio KCTC 2471 Growth Inhibition Represented by Alcohol Extracts

Growth inhibition ring diameter (mm) Escherichia coli E. Aerogenes Pneumococcal bacillus Pseudomonas aeruginosa JUTIX Enteritis Vibrio One* 5 10 One 5 10 One 5 10 One 5 10 One 5 10 One 5 10 Alder -** 12 16 - - - - - 10 - - - - - - - 10 12 Tangerine - - - - - - - - - - - - - - - - - - licorice - - - - - - - - - - - - - - - - - 12 Earth disease - - - - - - - - - - - - - - - - - - Gala - - - - - - - - - - - - - - - - - 11

* Concentration of extract of medicinal plants, mg / ml

** not measured

Table 7. Growth Inhibition of Wood Vinegar for Various Microorganisms

Growth inhibition ring diameter (mm) 5 * 15 25 35 45  B. Subtilis PM 125-** -**- - - - - M. Luteus KCTC 1056 - - - - - Staphylococcus aureus KCTC 1916 - - - - - MRSA CCARM 3115 - - - - -

MRSA CCARM 3089 - - - - - MRSA CCARM 3561 - - - - - Escherichia coli D 31 - - - 13 16 E. Aerogenes bacteria KCTC 2190 - - - - - Pneumococcal bacteria KCTC 2208 - - - - - Pseudomonas aeruginosa KCTC 2004 - - - - - Juventus KCTC 1925 - - - - - Enteritis Vibrio KCTC 2471 - 9 11 14 18

Volume of wood vinegar, μl

** not measured

After inoculation into the activated whole culture TSB medium, and incubated until the logarithmic growth of bacteria, inoculate and spread the bacterial solution on the TSA plate medium and 50 ml of five herbal medicine extract samples at a concentration of 1 ~ 10㎎ / ㎖ on the surface of the medium and The diameter of the inhibition zone around the paper disc against 6 Gram-positive bacteria, 5 Gram-negative bacteria, and marine bacteria, including three methicillin-resistant strains, S. aureus, was measured at a concentration of 5-45 μl of wood vinegar. The results are shown in Table 5.

The tangerine extract did not show antimicrobial activity against 12 strains at all concentrations of 1-10 mg / ml (Tables 5 and 6). A study on the antimicrobial activity of tangerine in this experiment was conducted in the previous study (Cho et al., 2003; Choi et al., 2002). This is in good agreement with the report of no antimicrobial activity against S. gallinarum and similar to the results of studies showing that the MIC of naringin, a large amount of antimicrobial substance present in tangerines, is greater than 1 mg / ml (Han and You, 1988) Indicates.

The antimicrobial activity against the earth disease extract shown in Tables 5 to 6 and FIG. 1 did not show antimicrobial activity against all strains tested at a concentration of 1 mg / ml. At a concentration of 5 mg / ml, growth inhibition ring of 10 mm was formed in MRSA CCARM 3115, but antimicrobial activity was not observed in other strains. At a concentration of 10 mg / ml, 10 mm growth inhibition ring was formed for MRAS CCARM 3809. However, antimicrobial activity was not observed for the other nine strains.

The antimicrobial activity of the browning extracts shown in Tables 5, 6, 1 and 2 did not show antimicrobial activity against all 12 strains at a concentration of 1 mg / ml. This result is similar to the previous study (Shin et al., 1997) where no antimicrobial activity was observed for 9 test strains at a concentration of 1.3 mg / ml for 75% ethanol extract of browning. In the case of the concentration of 5 mg / ㎖ of the gallium extract showed an antimicrobial activity to form a growth inhibitory ring of 12 mm, 12 mm in MRAS CCARM 3809 and 3115, respectively, no antimicrobial activity was observed for other strains. At a concentration of 10 mg / ml, growth inhibition rings of 15 mm and 14 mm were formed for MRAS CCARM 3809 and 3115, respectively, and the antimicrobial activity increased as the concentration increased, and it was 11 for marine bacterium Vibrio KCTC 2471. It showed the antimicrobial activity of mm (Fig. 2).

Licorice extract was not observed antimicrobial activity at a concentration of 1 mg / ㎖ (Tables 5 and 6). At 5 mg / ml, MRAS CCARM 3115, 3809, and 3561 showed antimicrobial activity forming growth inhibitory rings of 13 mm, 13 mm, and 11 mm, respectively, and 12 mm against marine bacteria enteritis Vibrio KCTC 2471. It showed the antimicrobial activity of (Table 5 and 6). At the concentration of 10 mg / ml, licorice extract showed growth inhibition ring of 10 mm for gram-positive bacillus subtilis PM 125 and growth inhibition ring of 16, 16 and 14 mm for MRSA CCARM 3115, 3809 and 3561, respectively. 1 and Table 7, and showed an antimicrobial activity forming a growth inhibition ring of 12 mm against enteritis Vibrio KCTC 2471 (Fig. 2 and Table 6). As shown in this study, licorice extract showed relatively strong antimicrobial activity against Gram-positive bacteria, negative bacteria and marine bacteria. Liguiritigenin was identified from 75% ethanol extract of licorice. In addition, it is similar to the report that a wide range of antimicrobial activity was observed for various strains (Ahn et al., 1998; Lee et al., 2002; Shin et al., 1994). However, previous studies (Cai et al., 2002) reported that hot water and ethanol extract of licorice formed growth inhibitory rings at 200 mg / ml against S. aureus and Escherichia coli 0157: H7. Stronger antimicrobial activity was reported, which is thought to be due to differences in the extraction conditions, methods and species.

The antimicrobial activity of the algae extract showing the strongest activity as a result of the inhibition of growth inhibition is shown in Tables 6 and 7 and FIGS. 1 and 2. Growth inhibitory rings of 15, 12 and 10 mm were formed for MRSA CCARM 3115, 3809 and 3561 at a concentration of 1 mg / ml. At the concentration of 5 mg / mL, the algae extract formed a 10 mm growth inhibitory ring against the Gram-positive bacteria M. luteus KCTC 1056, and 21, 19, and 12 mm growth inhibition for MRAS CCARM 3115, 3809, and 3561, respectively. A ring was formed. The growth inhibitory ring of 12 mm was formed in the Gram-negative bacterium Escherichia coli D 31 at a concentration of 5 mg / ml, and had an antibacterial activity of 10 mm against the marine bacterium enteritis Vibrio KCTC 2471. At 10 mg / ml, antimicrobial activity against Bacillus subtilis PM 125 and M. luteus KCTC 1056 was observed as antimicrobial activity against Gram-positive bacteria. MRAS CCARM 3115 , 3809 and 3561, strong antimicrobial activity was observed to form a growth inhibitory ring of 24, 20, 15 mm, respectively (Fig. 1). In addition, the antimicrobial activity against Gram-negative bacteria at a concentration of 10 mg / ml resulted in growth inhibition rings of 16 and 10 mm, respectively, against E. coli D 31 and K. pneumoniae KCTC 2208, and against enteritis Vibrio bacteria KCTC2471. It showed an antimicrobial activity of 12 mm (Fig. 2). Despite the strong antimicrobial activity of alders, there is no report on the antimicrobial activity of algae. Therefore, the research results presented in this study suggest that alderwood is a good material for the development of natural antimicrobial agents.

The antimicrobial activity of wood vinegar is shown in Table 8 and FIG. Wood grass liquor formed 13 and 16 mm growth inhibitory rings at 35 and 45 μl against E. coli D 31 and 15 μl to 9 mm growth inhibitory ring against enteritis vibriovirus KTTC 2471. As it increased, antimicrobial activity also increased. However, compared with previous studies (Lee et al., 2004), the antibacterial activity of the wood vinegar tested in this study was lower than that of the previous study. I think this is a similar result to the lower one.

Looking at the antimicrobial activity of the medicinal herbs used for the preparation of hangover, the antimicrobial activity of the medicinal herbs used in this study showed a wide range of antimicrobial activity against gram-negative and positive bacteria. Rather, it showed strong antimicrobial activity. In addition, as a result of showing strong antimicrobial activity against S. aureus resistant to methicillin, which was developed as an alternative antibiotic for penicillin, natural herbal medicines are useful for developing strong antibiotics against antibiotic resistant strains. It will be a resource that suggests availability as a resource. It is also useful for the development of beverages made of natural preparations without the addition of sodium benzoate, which is a synthetic preservative generally used in the manufacture of beverages, at a time when the retardation phenomenon of synthetic products has become prominent due to the economic improvement and advancement of living standards. I think it will be material.

Test Example 4  : Hemolytic activity ( Hemolysis ) Measure

Human and rat red blood cells were used to measure hemolytic activity (Park et al., 1996). Fresh ring erythrocytes were washed with 50 mM phosphate buffer (pH 7.4) containing 150 mM NaCl to adjust the concentration of the final erythrocytes to 5% (v / v). The erythrocyte solution and the extract sample were continuously added, followed by incubation at 37 ° C. for 1 hour, followed by centrifugation (8,000 × g, 10 min). The supernatant was measured at 450 nm using ELISA MQX200. The relative absorbance at the time of addition of the extracted sample was defined as 0% (A) and the absorbance when 1% Triton X-100 was added as 100% (B) when the red blood cells did not contain the extract sample. The hemolytic activity% was calculated from (C).

Hemolytic activity (%) = (C-A / B-A) × 100

Taken together, the hangover drink prepared in this study is not only excellent in ADH inhibitory activity in vivo, but also a hangover drink with strong antioxidant activity.

Hangover beverage composition of the present invention can implement the effect of inhibiting the liver protection effect or hangover phenomenon by inhibiting the production of acetaldehyde which is an oxide of alcohol by inhibiting the activity of alcohol dehydrogenase.

Claims (3)

A hangover relieving drink composition comprising alnus japonica, citrus unshiu, licorice (Glycyrrhiza uralensis), earth disease (Hovenia dulcis), and herbal extract extracted from Pueraria thunbergiana with ethyl alcohol as an active ingredient. The method according to claim 2, wherein the beverage composition further comprises one or more selected from honey, oligosaccharides and citric acid, a herbal composition for relieving hangover as an active ingredient. Herbal medicine comprising extracting dried herbal medicines of licorice, brown flower, alderwood, tangerine and earth disease with ethyl alcohol, filtering the extracted material with a nonwoven fabric, concentrating under reduced pressure and centrifuging, and then freezing and drying the obtained supernatant. Method for producing a hangover relief composition comprising the extract as an active ingredient.

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