A method for producing fermented Kenponashi fruit having hangover relieving and hepatoprotective effects, and a beverage composition produced using the fruit as the main ingredient

Abstract

The present invention relates to a method for producing fermented vinegar from Kenponashi fruit, which has hangover relieving and liver-protecting effects, and a beverage composition produced using the same as a main ingredient. The method for producing fermented vinegar from Kenponashi fruit of the present invention involves preparing a Kenponashi fruit concentrate using Kenponashi fruit, adjusting the concentrate to 16°Brix, inoculating it with 10% (v / v) yeast starter, and subjecting it to alcoholic fermentation at a temperature of 28-32°C, followed by filtration, and using the resulting alcoholic fermentation filtrate as the acetic acid fermentation substrate. The acetic acid fermentation is carried out by inoculating the fermentation liquid obtained by alcoholic fermentation of Kenponashi fruit with 10% (v / v) seed vinegar, followed by fermentation at a temperature of 28-32°C and 120-180 rpm.

Description

[Technical field]

[0001] The present invention relates to a method for producing fermented Kenponashi fruit having hangover-relieving and hepatoprotective effects, and a beverage composition produced using the same as a main ingredient, and more specifically, to a method for producing a fermented vinegar having hangover-relieving and hepatoprotective effects by fermenting Kenponashi fruit or Kenponashi fruit, and using the fermented vinegar to produce a beverage composition. [Background technology]

[0002] Hovenia dulcis Thunb is a deciduous tall tree of the Rhamnaceae family, also known as Hovenia dulcis, Jiku, or Broad-leaved Hovenia, and is known in traditional Chinese medicine for its excellent effects in improving liver function and detoxifying alcohol.

[0003] In botanical illustrations and herbalism, the fruit of the Kenponashi tree is known to be excellent for diuresis, thirst relief, liver function protection and alcohol toxin removal. In recent years, the pharmacological effects of the fruit, such as antidiabetic and antioxidant effects, promotion of alcohol metabolism in the body, hangover relief and thirst relief, have become known, and various efforts are underway to use this fruit in the production of health functional foods.

[0004] Moderate alcohol intake is beneficial for the human body in promoting health and promoting social interaction, but excessive drinking can lead to hangovers, vomiting, dry throat, muscle pain, headaches and nausea, disrupting daily life, and continued alcohol intake can cause metabolic disorders and inhibit the absorption of nutrients, leading to nutritional deficiencies, alcoholic fatty liver, cirrhosis, hepatitis and other liver diseases, as well as diabetes, neuropsychiatric disorders and cancer.

[0005] Alcohol is primarily oxidized in the liver through metabolic processes, first to acetaldehyde and then to acetate. However, acetaldehyde, the substance that causes hangovers in the body, can damage liver cells, and excess alcohol affects lipid metabolism, resulting in the overproduction of fatty acids and reduced fatty acid oxidation, which can lead to fatty liver. Therefore, research is underway to relieve hangovers and prevent liver damage caused by alcohol intake.

[0006] In recent years, due to changes in the nature of illnesses, consumers have become increasingly interested in slofoods, which are health foods such as fermented foods and traditional foods, and in hangover relief drinks made with Kenponashi.

[0007] Vinegar, along with alcohol, is a representative fermented food with a long history in the human diet, containing sourness, sugars, volatile and non-volatile organic acids, esters, amino acids, etc., and the components of vinegar have been reported to have physiologically active functions such as fatigue recovery and anti-aging, as well as efficacy in treating high blood pressure, arteriosclerosis, and regulating internal metabolism, and are therefore attracting attention as a functional health food. Vinegar is classified into brewed vinegar, which is made from fruits and grains as main ingredients, and synthetic vinegar, which is made by diluting acetic acid or glacial acetic acid with water.

[0008] Recently, with the improvement of dietary culture and growing awareness of health, research has been actively conducted on natural vinegar made from various fruits and vegetables such as apples, persimmons, and grapes rather than synthetic vinegars. However, research on vinegar made from Kenponashi fruit for hangover relief and liver protection has been insufficient. Summary of the Invention [Problem to be solved by the invention]

[0009] The present invention has been made to solve the above-mentioned problems, and its object is to develop an acetic acid fermentation concentrate using the fruit of the Kenponashi tree, which has been reported to be excellent for improving liver function and decomposing alcohol, as a functional food ingredient, and to provide a functional beverage that has the effect of relieving hangovers and protecting liver function. [Means for solving the problem]

[0010] In order to solve the above technical problems, the present invention uses Kenponashi fruit to produce fermented vinegar, which is then used to produce a functional beverage having hangover relieving and hepatoprotective effects.

[0011] The method for producing fermented vinegar using Kenponashi fruits of the present invention involves preparing a Kenponashi fruit concentrate using Kenponashi fruits, adjusting this to 16°Brix, inoculating it with 10% (v / v) yeast starter, and subjecting it to alcoholic fermentation at a temperature of 28-32°C, followed by filtration, and using the resulting alcoholic fermentation filtrate as the acetic acid fermentation substrate. The acetic acid fermentation is carried out by inoculating the fermentation liquid obtained by alcoholic fermentation of Kenponashi fruits with 10% (v / v) seed vinegar, followed by fermentation at a temperature of 28-32°C and 120-180 rpm.

[0012] The concentrated solution of Kenponashi fruit can be prepared by adding 5 to 10 times the weight of washed Kenponashi fruit to water, heating the fruit, extracting it with hot water, and concentrating the extract, or by extracting it with alcohol.

[0013] For the alcoholic fermentation, the strain Saccharomyces cerevisiae (KCCM11306) was subcultured on a YM plate medium and then statically cultured in a YM liquid medium for 48 hours.

[0014] The YM plate medium was prepared by diluting the fermentation strain with 0.85% NaCl solution, smearing it on LB, YM, PDA, or MRS plate medium, culturing it in an incubator at 28 to 32°C, and then culturing it in liquid medium.

[0015] The YM liquid medium contains 100 parts by weight of water, 0.5 to 1.5 parts by weight of dextrose, 0.2 to 0.8 parts by weight of peptone, 0.1 to 0.5 parts by weight of yeast extract, and 0.1 to 0.5 parts by weight of malt extract.

[0016] The acetic acid fermentation is carried out by culturing Acetobacter aceti (KCCM12654) in a liquid medium prepared by mixing 100 parts by weight of water with 2 to 3 parts by weight of mannitol, 0.1 to 0.5 parts by weight of peptone, and 0.3 to 0.8 parts by weight of yeast extract at 30° C. for 48 hours.

[0017] The beverage composition using Kenponashi fruit fermented vinegar of the present invention is produced by containing 100 parts by weight of purified water, 0.5 to 7 parts by weight of Kenponashi fruit fermented vinegar, 1 to 3 parts by weight of Kenponashi concentrate, 10 to 15 parts by weight of isomerized sugar, 1 to 5 parts by weight of fructooligosaccharides, 0.5 to 1 part by weight of apple concentrate, 0.1 to 0.2 parts by weight of anhydrous citric acid, 0.1 part by weight of yeast extract powder, 0.01 to 0.03 parts by weight of complex Scutellaria baicalensis extract, 1 to 2 parts by weight of taurine, 0.01 to 0.10 parts by weight of milk thistle extract powder, and 0.01 to 0.05 parts by weight of curcumin. Effect of the Invention

[0018] The present invention has the effect of providing a fermented vinegar using Kenponashi fruit, which is excellent in improving liver function and decomposing alcohol.

[0019] In addition, the present invention can provide a functional food ingredient, such as a hangover relief beverage, by using the acetic acid fermentation liquid containing the fermented vinegar as a main ingredient, and thus has the effect of providing a functional beverage having liver function protecting properties. BEST MODE FOR CARRYING OUT THEINVENTION

[0020] The method for producing fermented vinegar using Kenponashi fruits of the present invention involves preparing a Kenponashi fruit concentrate using Kenponashi fruits, adjusting this to 16°Brix, inoculating it with 10% (v / v) yeast starter, and subjecting it to alcoholic fermentation at a temperature of 28-32°C, followed by filtration, and using the resulting alcoholic fermentation filtrate as the acetic acid fermentation substrate. The acetic acid fermentation is carried out by inoculating the fermentation liquid obtained by alcoholic fermentation of Kenponashi fruits with 10% (v / v) seed vinegar, followed by fermentation at a temperature of 28-32°C and 120-180 rpm. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0021] The present invention will be described in detail below with reference to examples to help the understanding of the present invention. However, the examples according to the present invention can be modified into various other forms, and the scope of the present invention should not be construed as being limited to the following examples. The examples of the present invention are provided to more completely explain the present invention to those having ordinary skill in the art.

[0022] "Fermented Kenponashi" as used throughout the specification of this invention is defined as meaning when the inventor ferments Kenponashi or Kenponashi fruit to produce "fermented vinegar", and more precisely, fermented Kenponashi means "fermented vinegar from Kenponashi fruit".

[0023] The method for producing fermented vinegar using Kenponashi fruit of the present invention involves preparing a Kenponashi fruit concentrate using Kenponashi fruit, adjusting the concentrate to 16°Brix, inoculating 10% (v / v) yeast starter and subjecting it to alcoholic fermentation at a temperature of 28-32°C, filtering the alcoholic fermentation filtrate, and using the resultant filtrate as the acetic acid fermentation substrate. The sugar content of the Kenponashi fruit concentrate is adjusted to 14-18°Brix with a saccharification solution, preferably 16°Brix.

[0024] The starter is prepared by inoculating the alcohol-fermenting bacteria Saccharomyces cerevisiae (KCCM11306) into a concentrated liquid of Kenponashi fruit adjusted to 14-16°Brix, and culturing it at a temperature of 28-32°C for 40-56 hours.

[0025] The acetic acid fermentation is carried out by inoculating the fermented liquid obtained by alcoholic fermentation of Kenponashi fruit with 10% (v / v) seed vinegar, followed by fermentation at 120 to 180 rpm at a temperature of 28 to 32° C. The seed vinegar is prepared by filtering the alcoholic fermented liquid of Kenponashi fruit obtained by alcoholic fermentation, inoculating the filtrate with acetic acid bacteria strain Acetobacter aceti (KCCM deposited strain: 12654), and culturing for 48 hours at a temperature of 28 to 32° C. and a rotation speed of 120 to 180 rpm, preferably 150 rpm, and culturing the resulting mixture for use as the seed vinegar.

[0026] The Kenponashi fruit concentrate can be prepared by adding 5 to 10 times the weight of washed Kenponashi fruit to water, heating the fruit, extracting it with hot water, and then concentrating the extract or extracting it with alcohol.

[0027] For the alcoholic fermentation, the strain Saccharomyces cerevisiae (KCCM11306) was subcultured on a YM plate medium and then statically cultured in a YM liquid medium for 48 hours.

[0028] The YM plate medium was prepared by diluting the fermentation strain with 0.85% NaCl solution, smearing it on LB, YM, PDA, or MRS plate medium, culturing it in an incubator at 28 to 32°C, and then culturing it in liquid medium.

[0029] The YM liquid medium contains 100 parts by weight of water, 0.5 to 1.5 parts by weight of dextrose, 0.2 to 0.8 parts by weight of peptone, 0.1 to 0.5 parts by weight of yeast extract, and 0.1 to 0.5 parts by weight of malt extract.

[0030] The acetic acid fermentation is carried out by culturing Acetobacter aceti (KCCM12654) in a liquid medium prepared by mixing 100 parts by weight of water with 2 to 3 parts by weight of mannitol, 0.1 to 0.5 parts by weight of peptone, and 0.3 to 0.8 parts by weight of yeast extract at 30° C. for 48 hours.

[0031] The beverage composition using Kenponashi fruit fermented vinegar of the present invention is produced by containing 100 parts by weight of purified water, 0.5 to 7 parts by weight of Kenponashi fruit fermented vinegar, 1 to 3 parts by weight of Kenponashi concentrate, 10 to 15 parts by weight of isomerized sugar, 1 to 5 parts by weight of fructooligosaccharides, 0.5 to 1 part by weight of apple concentrate, 0.1 to 0.2 parts by weight of anhydrous citric acid, 0.1 part by weight of yeast extract powder, 0.01 to 0.03 parts by weight of complex Scutellaria baicalensis extract, 1 to 2 parts by weight of taurine, 0.01 to 0.10 parts by weight of milk thistle extract powder, and 0.01 to 0.05 parts by weight of curcumin.

[0032] The silymarin contained in milk thistle (Silybum marianum L. Gaertner) is a flavonolignan and actually contains a mixture of four isomers, namely silybin, isosilybin, silydianin and silychristin, which have the same molecular formula of C25H22O10.

[0033] Silymarin has been reported to inhibit the proliferation of cancer cells and to be effective against diabetes. It also has antimicrobial activity against gram-positive bacteria. Recently, it has also been reported to be effective against skin damage caused by radiation and ultraviolet rays. It has also been reported to have a hepatoprotective effect, prevent alcohol-induced lipid oxidation, and protect against alcoholic cirrhosis of the liver.

[0034] Curcumin is a type of polyphenol found primarily in Indian turmeric. Curcumin is known to give turmeric its yellow color.

[0035] Curcumin is a food additive that is used to color food with a slight yellow color, and its E code is E100. It has been revealed that curcumin and a specific component related to curcumin, PPAR-gamma (Peroxisome Proliferator Activated Receptor-gamma), act as a base to block mediators that cause inflammation, and have anti-inflammatory, antioxidant and antibacterial effects. Other ingredients, including yeast extract powder and complex Scutellaria root extract, are purchased from the market and mixed for use. 1. Acute alcohol administration animal experiment design

[0036] In the present invention, 48 male Sprague-Dawley rats were provided by Hyochang Science (Daegu, Korea) as experimental animals. The animals were kept in an animal breeding room with a constant temperature (22±2°C) and humidity (50±5%) under a 12-hour light-dark cycle. For acute alcohol decomposition by egg mass method, the rats were divided into an alcohol only control group (Control), a 1.5% concentrated Kenponashi fruit group (HT), a 4% diluted Kenponashi fruit fermented vinegar group (PHT), a Kenponashi hangover relief drink group (DHD), and positive control groups (A, B).

[0037] For the positive control group, a commercially available product from another company that is marketed as a hangover relief drink was used. Two animals from each group were separated into acrylic cages and used after adapting to solid food for one week. On the day of the experiment, after fasting for 15 hours, the rats were administered 7ml / kg of each sample, and 30 minutes later, 5ml / kg of 40% fermented alcohol was orally administered, and blood was collected 180 minutes later to measure the alcohol and acetaldehyde concentrations. The collected blood was left at room temperature for 3 hours, then centrifuged at 12,000 rpm for 10 minutes, and only the supernatant was used for the experiment. 2. Measurement of blood alcohol and acetaldehyde levels

[0038] Serum alcohol concentrations were measured using an ethanol assay kit (Megazyme, Ireland), and serum acetaldehyde concentrations were measured using an acetaldehyde assay kit (Megazyme, Ireland). 3. Experimental Results A) Measurement of alcohol and acetaldehyde concentrations in an acute alcoholic rat model

[0039] The results of the alcohol decomposition ability of the acute alcoholic rat model are as follows. Alcohol and acetaldehyde concentrations were highest in the control group, and similar concentrations were confirmed in the HT group. The PHT group showed a significant decrease in blood alcohol and acetaldehyde concentrations when compared to the control and HT groups, and the DHD group was confirmed to have a greater decrease than the positive control groups A and B, and was particularly significantly different from group A. Ingestion of the Kenponashi fermented vinegar drink is judged to be excellent for alcohol decomposition in the body, as it reduced blood alcohol and acetaldehyde concentrations.

[0040] TIFF2024545732000001.tif115166

[0041] TIFF2024545732000002.tif108166

[0042] Figure: Effect of DHD on (A) serum alcohol concentrations and (B) serum acetaldehyde concentrations. Data values ​​with different superscripts indicate significant differences (p<0.05) by Duncan's multiple range test.

[0043] 4. Confirmation of hangover relief and hepatoprotective effects in a chronic alcohol administration model of a fermented vinegar drink made from Kenponashi fruit, which is the main ingredient B. Materials and methods A) Chronic alcohol administration animal study design

[0044] The experimental animals were 48 male Sprague-Dawley rats provided by Hyochang Science (Daegu, Korea), and were divided into six groups: normal group, control group administered with alcohol alone, 1.5% concentrated solution of Kenponashi fruit group, 4% diluted solution of processed Kenponashi fruit group, Kenponashi hangover relief drink group, and positive control group administered with silymarin, known to improve liver function. Animals in each group were separated into 2 groups in acrylic cages by egg mass method, and the experiment was carried out after adapting to solid food for one week. To prevent acute toxicity from alcohol, 10 and 20% fermented alcoholic alcohol was administered for one week, and each sample was administered at the same time as 30% fermented alcoholic alcohol was administered for four weeks. The positive control group was administered 50mg / kg, the remaining groups except the positive control group were administered 7ml / kg, and the normal group and control group were administered the same amount of distilled water. On the day of the experiment, the animals were fasted for 15 hours, then sacrificed and their blood and organs were collected. The collected blood was left at room temperature for 3 hours, then centrifuged at 12000rpm for 10 minutes, and only the supernatant was used for the experiment.

[0045] The basic diet used in this experiment was solid diet purchased from a supplier, and water or alcohol and food were provided ad libitum. Body weight, food intake, and water or alcohol intake were measured daily. Animal experiments were conducted ethically in accordance with the experimental animal guidelines of Dong-A University.

[0046] B) Measurement of blood alcohol and acetaldehyde concentrations Serum alcohol concentrations were measured using an ethanol assay kit (Megazyme, Ireland), and serum acetaldehyde concentrations were measured using an acetaldehyde assay kit (Megazyme, Ireland).

[0047] C) Measurement of liver tissue alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activities

[0048] 100 mg of liver tissue was mixed with 500 μl of 1x PBS and homogenized, then centrifuged at 10,000 rpm, 4°C, for 5 minutes, and the supernatant was used for the experiment. ADH activity was measured using an ADH activity kit (Bioassay systems, San Francisco Bay), and ALDH activity was measured using an ALDH analysis kit (BioVision, California).

[0049] D) Measurement of lipid peroxide content Lipid peroxides were measured by measuring the content of thiobarbituric acid reactive substances (TBARS). 1ml of the homogenized liver tissue solution was mixed with 2ml of TBA (2-thiobarbituric acid) reagent and then reacted in boiling water for 30 minutes. The reaction was then stopped with ice water and centrifuged at 3000 rpm for 10 minutes, and the supernatant was collected and measured for absorbance at 535 nm.

[0050] E) Analysis of lipid concentrations and biochemical indicators Serum LDL and HDL cholesterol, triglyceride content, ALT (GPT), AST (GOT), and GGT (γ-GTP) concentrations were analyzed at the Southeast Medi-Chem Institute (Korea), a specialized analytical institution, and blood total cholesterol content and liver tissue triglyceride concentration were measured using analytical kits (Elabscience, United States).

[0051] F) Glutathione content analysis Glutathione content analysis of serum and liver tissue was measured using a kit (Elabscience, United States).

[0052] G) Hematoxylin and eosin (H&E) staining analysis The liver tissue was fixed with 4% formaldehyde solution, stained with H&E solution, and observed under an optical microscope.

[0053] B. Experimental results A) Changes in body weight and beverage intake in a chronic alcohol-induced rat model The results of changes in body weight and beverage intake of chronic alcohol-administered animals are shown in the table below. The initial body weight of rats was not significantly different in any group, but the final body weight was significantly different between the Control group and the Normal group. The HT group, PHT group, and Silymarin group were not significantly different from the Control group, but the PHT group rats showed an increase in body weight. The DHD group was not significantly different from the Normal group. No significant differences were observed in beverage intake in any group.

[0054] Table: Effects of DHD on body weight, body weight gain, and water consumption in chronically ethanol-treated rats. TIFF2024545732000003.tif83166

[0055] Data values ​​are presented as mean ± SEM (n = 6). Data values ​​with different superscripts indicate significant differences (p < 0.05) by Duncan's multiple range test.

[0056] B) Changes in organ weights in a chronic alcoholic rat model The changes in organ weights in chronic alcohol-administered rats are shown in the table below. The weights of the spleen, kidney, and heart did not show significant differences in any of the groups. The liver weight was highest in the control group, while the normal group showed the lowest weight. It was confirmed that the liver weights of the HT, PHT, and Silymarin groups were significantly decreased compared to the control group, and that the liver weight of the PHT group was lower than that of the HT and Silymarin groups. The DHD group showed lower weights than the HT, PHT, and Silymarin groups, and was not significantly different from the normal group. The testis weight was highest in the control group, while the PHT group showed lower weights than the HT and Silymarin groups. The DHD group showed lower testis weights, with no significant difference from the normal group. It is considered that the intake of fermented vinegar drink of Kenponashi fruit suppresses alcoholic fatty liver. Table: Changes in DHD in relation to organ weights in chronically ethanol-administered rats TIFF2024545732000004.tif96166 Data values ​​are shown as mean ± SEM (n = 6). Data values ​​with different superscripts indicate significant differences (p < 0.05) by Duncan's multiple range test. B) Changes in liver damage indicators in a chronic alcoholic rat model

[0057] The results of measuring liver damage indicators in rats chronically administered alcohol are shown in the figure below. The GOT, GPT and γ-GTP concentrations were shown to have increased the most in the control group, and were lowest in the normal group. The GOT, GPT and γ-GTP concentrations in the PHT group were shown to have decreased significantly compared to the control, HT and Silymarin groups. The GOT, GPT and γ-GTP concentrations in the DHD group were not significantly different from the normal group, and were shown to have been significantly lower than the control, HT, PHT and Silymarin groups. It is considered that the intake of Kenponashi fermented vinegar drink protects the liver from alcoholic liver damage.

[0058] TIFF2024545732000005.tif108166

[0059] TIFF2024545732000006.tif108166

[0060] TIFF2024545732000007.tif108166

[0061] Figure: Effects of DHD on (A) serum GOT (AST) content, (B) serum GPT (ALT) content, and (C) serum γ-GTP content in chronic ethanol-administered rats.

[0062] Data values ​​are presented as mean ± SEM (n = 6). Data values ​​with different superscripts indicate significant differences (p < 0.05) by Duncan's multiple range test.

[0063] C) Changes in alcohol decomposition enzyme activity (ADH, ALDH) in a chronic alcoholic rat model

[0064] The results of ADH and ALDH activity in chronic alcohol-administered rats are as follows. ADH activity was higher in the HT, PHT, DHD, and Silymarin groups than in the control group, and was significantly increased in the Silymarin, PHT, and DHD groups. The PHT group was significantly increased compared to the HT and Silymarin groups, and the DHD group showed significantly the highest ADH activity. ALDH activity was higher in the PHT group than in the control, HT, and Silymarin groups, and was significantly higher than the Silymarin group. The DHD group showed significantly the highest ALDH activity. It is considered that the intake of Kenponashi fermented vinegar drink increases the activity of alcohol-decomposing enzymes.

[0065] TIFF2024545732000008.tif102166

[0066] TIFF2024545732000009.tif102166

[0067] Figure: Effects of DHD on intrahepatic (A) ADH activity and (B) ALDH activity in chronic ethanol-administered rats.

[0068] Data values ​​are presented as mean ± SEM (n = 6). Data values ​​with different superscripts indicate significant differences (p < 0.05) by Duncan's multiple range test.

[0069] D) Changes in blood alcohol and acetaldehyde concentrations in a chronic alcoholic rat model The results of alcohol and acetaldehyde concentrations in chronic alcohol-administered rats are shown in the figure below. Blood alcohol concentrations were significantly higher in the Control group and lower in the Normal group. The PHT group showed significantly lower concentrations than the HT group, and the DHD group showed significantly lower concentrations than the HT, PHT, and Silymarin groups, and showed no significant difference from the Normal group. The acetaldehyde concentration was significantly higher in the Control group and lower in the Normal group. The PHT group showed significantly lower concentrations than the HT and Silymarin groups, and the DHD group showed significantly lower acetaldehyde concentrations than the HT, PHT, and Silymarin groups. The DHD group also showed no significant difference from the Normal group. It is judged that the intake of Kenponashi fermented vinegar drink reduces blood alcohol and acetaldehyde concentrations.

[0070] TIFF2024545732000010.tif102166

[0071] TIFF2024545732000011.tif102166

[0072] Figure: Effects of DHD on (A) serum alcohol concentrations and (B) serum acetaldehyde concentrations in chronically ethanol-treated rats.

[0073] Data values ​​are presented as mean ± SEM (n = 6). Data values ​​with different superscripts indicate significant differences (p < 0.05) by Duncan's multiple range test.

[0074] E) Changes in liver and blood glutathione (GSH) concentrations in a chronic alcoholic rat model The results of measuring GSH concentrations in the liver and blood of chronic alcohol-administered rats are shown in the figure below. The control group showed the lowest GSH concentrations in the liver tissue and blood, the HT group showed higher concentrations than the control group but the difference was not significant, and the PHT group showed higher concentrations than the HT group but the difference was not significant. The DHD group showed significantly higher GSH concentrations in the liver and blood. It is considered that the intake of the Kenponashi fermented vinegar drink activates GSH in the liver and blood.

[0075] TIFF2024545732000012.tif102166

[0076] TIFF2024545732000013.tif102166

[0077] Figure: Effect of DHD on (A) hepatic GSH content and (B) serum GSH content in chronic ethanol-administered rats.

[0078] Data values ​​are presented as mean ± SEM (n = 6). Data values ​​with different superscripts indicate significant differences (p < 0.05) by Duncan's multiple range test.

[0079] F) Changes in liver and blood TBARS (MDA) concentrations in a chronic alcoholic rat model The liver and blood lipid peroxide concentrations in chronic alcoholic rats are shown in the figure below. The liver lipid peroxide concentration was highest in the control group and lowest in the normal group. The HT group had a lower lipid peroxide concentration than the control group, but the difference was not significant. The PHT group showed a significantly lower concentration than the HT group and a lower concentration than the Silymarin group. The DHD group showed a significantly lower lipid peroxide concentration than the PHT group and no significant difference from the normal group. The control group showed a significantly highest concentration of blood lipid peroxide. The HT and Silymarin groups were confirmed to have significantly lower concentrations than the control group, and the PHT group showed a significantly lower concentration than the HT and Silymarin groups. The DHD group showed a significantly lower concentration than the PHT group and no significant difference from the normal group. It is considered that the intake of Kenponashi fermented vinegar drink suppresses the production of lipid peroxide in the liver and blood.

[0080] TIFF2024545732000014.tif102166

[0081] TIFF2024545732000015.tif102166

[0082] Figure: Effect of DHD on (A) hepatic and (B) serum TBARS concentrations in chronically ethanol-administered rats.

[0083] Data values ​​are presented as mean ± SEM (n = 6). Data values ​​with different superscripts indicate significant differences (p < 0.05) by Duncan's multiple range test.

[0084] G) Changes in liver and blood triglyceride concentrations in a chronic alcoholic rat model The results of measuring the liver and blood triglyceride concentrations in chronic alcoholic rats are shown in the figure below. The control group showed significantly higher liver and blood triglyceride concentrations. The HT group showed significantly lower concentrations than the control group and similar to the Silymarin group. The PHT group showed significantly lower liver and blood triglyceride concentrations than the HT and Silymarin groups, and the DHD group showed significantly lower concentrations than the PHT group and similar to the Normal group. It is considered that the intake of Kenponashi fermented vinegar drink suppresses the production of liver and blood triglycerides.

[0085] TIFF2024545732000016.tif102166

[0086] TIFF2024545732000017.tif102166

[0087] Figure: Effects of DHD on (A) hepatic triglyceride content and (B) serum triglyceride content in chronic ethanol-administered rats.

[0088] Data values ​​are presented as mean ± SEM (n = 6). Data values ​​with different superscripts indicate significant differences (p < 0.05) by Duncan's multiple range test.

[0089] H) Changes in blood total cholesterol, LDL-cholesterol, and HDL-cholesterol concentrations in a chronic alcoholic rat model The results of blood total cholesterol, LDL-cholesterol, and HDL-cholesterol concentrations in chronic alcohol-administered rats are shown in the figure below. The total cholesterol concentration did not show significant differences in the HT and Silymarin groups when compared with the control group. The PHT group was confirmed to have a significantly decreased value compared to the HT group, and the DHD group showed a significantly lower total cholesterol concentration than the PHT group, and showed values ​​similar to those of the Normal group. The blood LDL-cholesterol concentration was shown to be significantly the highest in the Control group, and the HT and Silymarin groups showed significantly lower concentrations than the Control group. The PHT group was shown to have a significantly lower value than the HT group, and the DHD group showed a significantly lower concentration than the PHT group. In particular, it was confirmed that the DHD group was not significantly different from the Normal group. The HDL-cholesterol concentration was shown to be the lowest in the Control group, and the HT and Silymarin groups showed no significant differences from the Control group. The PHT group showed no significant difference from the HT and Silymarin groups, but the HDL-cholesterol concentration was confirmed to have increased further, showing a similar concentration to the Normal group. The DHD group showed a significantly higher concentration than the Normal group. It was confirmed that the intake of the Kenponashi fermented vinegar drink reduced total cholesterol and LDL-cholesterol concentrations, and increased HDL-cholesterol concentrations.

[0090] TIFF2024545732000018.tif102166

[0091] TIFF2024545732000019.tif102166

[0092] TIFF2024545732000020.tif102166

[0093] Figure: Effects of DHD on (A) total cholesterol content, (B) serum LDL cholesterol content, and (C) serum HDL cholesterol content in chronically ethanol-treated rats.

[0094] Data values ​​are presented as mean ± SEM (n = 6). Data values ​​with different superscripts indicate significant differences (p < 0.05) by Duncan's multiple range test.

[0095] I) Morphological and H&E staining changes in liver tissue in a chronic alcoholic rat model The morphological and H&E staining results of liver tissue in chronic alcohol-administered rats are as follows. When fatty liver is formed by alcohol, the color of the liver changes from reddish brown to yellowish brown, but in the case of the control group, it was shown that the color changed to yellowish brown due to the formation of fatty liver. The PHT group was shown to be more reddish brown than the control and HT groups, and the DHD group was a dark reddish brown similar to the normal group. H&E staining results showed many lipid droplets in the control group. It was confirmed that the HT group had fewer lipid droplets than the control group, and the PHT group was found to have less lipid droplet formation than the HT group. The DHD group was shown to have almost no lipid droplets and showed a morphology similar to that of the normal group. It is considered that the intake of fermented vinegar drink of Kenponashi fruit suppresses alcoholic fatty liver.

[0096] TIFF2024545732000021.tif64166

[0097] Figure: Effects of DHD on morphological changes in liver tissue in chronic ethanol-administered rats.

[0098] TIFF2024545732000022.tif51166

[0099] Figure: Effect of DHD on liver tissue morphology H&E (magnification 400x) in chronic ethanol-administered rats

Claims

1. In a method for producing fermented vinegar from Kenponashi fruit, which has hangover relieving and liver protecting effects, The method for producing the fermented vinegar of Kenponashi fruit is to prepare a Kenponashi fruit concentrate using Kenponashi fruit, adjust the temperature to 16° Brix, inoculate the concentrate with 10% (v / v) yeast starter, and ferment the concentrate at a temperature of 28-32° C., and then filter the alcohol fermentation liquid filtrate, which is used as an acetic acid fermentation substrate. The acetic acid fermentation is carried out by adding an alcohol fermentation liquid of Kenponashi fruit, inoculating it with 10% (v / v) of seed vinegar, and fermenting it at a temperature of 28 to 32° C. and at 120 to 180 rpm. A method for producing fermented vinegar from Kenponashi fruit, which has hangover relieving and liver protecting effects.

2. The concentrated solution of the Kenponashi fruit is prepared by adding 5 to 10 times the weight of the washed Kenponashi fruit to water, heating the fruit, and extracting the fruit with hot water to concentrate it. A method for producing the fermented Kenponashi fruit vinegar described in claim 1, which has hangover relieving and liver protecting effects.

3. The alcoholic fermentation was carried out by subculturing the strain Saccharomyces cerevisiae (KCCM11306) on a YM plate medium, and then statically culturing the strain in a YM liquid medium for 48 hours. A method for producing the fermented Kenponashi fruit vinegar described in claim 1 or 2, which has hangover relieving and liver protecting effects.

4. The acetic acid fermentation was carried out by culturing Acetobacter aceti (KCCM12654) in a liquid medium containing 100 parts by weight of water, 2 to 3 parts by weight of mannitol, 0.1 to 0.5 parts by weight of peptone, and 0.3 to 0.8 parts by weight of yeast extract at 30° C. for 48 hours. A method for producing the fermented Kenponashi fruit vinegar described in claim 1 or 2, which has hangover relieving and liver protecting effects.

5. The beverage composition using fermented vinegar of Kenponashi fruit contains 100 parts by weight of purified water, 0.5 to 7 parts by weight of fermented vinegar of Kenponashi fruit, 1 to 3 parts by weight of concentrated Kenponashi fruit, 10 to 15 parts by weight of isomerized sugar, 1 to 5 parts by weight of fructooligosaccharide, 0.5 to 1 part by weight of concentrated apple juice, 0.1 to 0.2 parts by weight of anhydrous citric acid, 0.1 part by weight of yeast extract powder, 0.01 to 0.03 parts by weight of composite Scutellaria root extract, 1 to 2 parts by weight of taurine, 0.01 to 0.10 parts by weight of milk thistle extract powder, and 0.01 to 0.05 parts by weight of curcumin. A beverage composition using fermented vinegar of Kenponashi fruit.

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