KR20230090136A - Composition for preventing or relieving hangover, comprising buckwheat vinegar extract - Google Patents

Abstract

The present invention relates to a composition for preventing or relieving a hangover, comprising a buckwheat vinegar extract. The buckwheat vinegar extract according to one example significantly improves the activity of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH), and quickly decomposes alcohol and acetaldehyde in the body, thereby having an excellent effect of preventing or relieving hangover.

Description

Composition for preventing or relieving hangover, comprising buckwheat vinegar extract}

The present invention relates to a composition for preventing or relieving a hangover, comprising a buckwheat vinegar extract.

Consumption of alcohol (C ₂ H ₅ OH) is rapidly increasing as a means of socializing and relieving stress of modern people, but hangover symptoms such as nausea, vomiting, dizziness, lethargy, headache, muscle pain, etc. As a result, it interferes with gastrointestinal diseases and biorhythms, reduces cognitive function, and in severe cases, causes social and economic damage due to reduced work efficiency the next day (Swift and Davidson, 1998; Woo et al., 2021).

In general, about 10% of alcohol is excreted through lungs, urine, and sweat, and 90% is metabolized and oxidized in the liver (Cederbaum, 2012; Lieber, 1992). At this time, alcohol metabolism in the body is oxidized to acetaldehyde by alcohol dehydrogenase (ADH) in the cytosol, CYP2E1 (cytochrome P4502E1) in the endoplasmic reticulum, and catalase (CAT) in the peroxisome, and then to the mitochondria. It moves and is oxidized by non-specific aldehyde dehydrogenase (ALDH) to acetic acid (CH ₃ COOH), which is finally excreted as urine (H ₂ O) and carbon dioxide (CO ₂ ) (Lin and Li, 1998; Kee et al., 2003). Alcohol is known to have various effects on liver metabolism depending on the amount consumed. Excessive alcohol intake causes the accumulation of toxic acetaldehyde in the body, causing hangover symptoms and liver cell damage (Hawkins and Kalant, 1972; Lieber, 1992).

Modern people's awareness of the quality of life is changing due to the increase in income level and life span, and interest is shown in hangover relief drinks as health functional foods using natural products to recover liver fatigue caused by stress, aging, and drinking ( Na et al., 2018). Recently, studies on the antioxidant ability and hangover relieving efficacy of foods using natural products without side effects have been actively conducted. Plant herbal medicines (79.4%) were the most common main ingredients for hangover improvement, and pine, alder, cinnamon, raisin, and fermented soybean extracts have been reported (Yang et al., 2004).

Republic of Korea Patent No. 10-2310813

As a result of intensive research to identify natural substances having hangover preventing, improving, and/or relieving effects, the present inventors found that buckwheat vinegar extract (eg, buckwheat seed vinegar extract) effectively promotes alcohol and/or acetaldehyde decomposition. It was confirmed that the present invention was completed.

An object of the present invention is to provide a pharmaceutical composition for preventing or relieving hangover, containing buckwheat vinegar extract.

Another object of the present invention is to provide a health functional food composition for preventing or relieving hangover, including buckwheat vinegar extract.

Another object of the present invention is to provide a food composition for preventing or relieving hangover, containing buckwheat vinegar extract.

One aspect for achieving the above object is to provide a composition for preventing, improving and/or relieving a hangover (e.g., a pharmaceutical composition, a health functional food composition, and/or a composition for food) containing a buckwheat vinegar extract. is to provide

As used herein, "extract" refers to a preparation obtained by squeezing a crude drug into an appropriate leachate and concentrating the leachate by evaporating the leachate. These may be a crude product and/or a purified product. The buckwheat vinegar extract can be prepared using a general extraction method, separation and purification method known in the art.

According to one example, the buckwheat vinegar extract can be obtained by adding 5 to 20 times, 10 to 30 times, 15 to 25 times, or 5 times to 30 times more vinegar than buckwheat by weight, and after extraction It may be further purified by methods such as fractionation, separation, and filtration. According to one example, the buckwheat vinegar extract is immersed in vinegar for 1 to 31 days, 10 to 20 days, or 14 to 21 days at a temperature condition of 15 to 25 ° C. After that, it can be obtained by removing the precipitate using a filter.

In one example, the buckwheat vinegar extract may include the following three steps:

Step 1 (steaming): buckwheat raw material is soaked in water for 30 to 40 minutes and then steamed at about 100° C. for 30 to 40 minutes;

Step 2 (drying): drying the steamed buckwheat raw material in the above step at 40 ± 2 ° C for 3-4 days (moisture content less than 13.5%); and

Step 3 (extraction):

Prepared with fermented buckwheat vinegar by adding vinegar (e.g., brown rice vinegar) in an amount of 10 to 30 times the weight of raw grain at 20 ± 5 ° C and extracting (or maintaining) for about 2 to 3 weeks, or

10-30 times the weight of raw grain Vinegar (e.g., brown rice vinegar) After adding sugar, floral ingredients (e.g., buckwheat sprouts, rose flower, marigold flower, and/or hibiscus flower) are blended (e.g. For example, buckwheat vinegar extract (buckwheat vinegar extract containing flower dimension; for example, buckwheat Process for preparing buckwheat vinegar, buckwheat whole vinegar, buckwheat marigold vinegar, buckwheat rose vinegar, and/or buckwheat hibiscus vinegar).

According to one example, the buckwheat vinegar extract may be obtained by acetic acid fermentation of buckwheat (eg, buckwheat seeds, specifically, bitter buckwheat seeds), for example, saccharification, alcohol fermentation and / or acetic acid fermentation It can be manufactured through the process.

Vinegar used to prepare the buckwheat vinegar extract according to one embodiment is not limited as long as it is commercially available, for example, synthetic vinegar made by a chemical method and / or natural It may be a fermented brewing vinegar (eg, rice vinegar, brown rice vinegar, grain vinegar, malt vinegar, apple vinegar, grape vinegar, red wine vinegar, persimmon vinegar, balsamic vinegar, and/or herbal vinegar).

In one example, the buckwheat vinegar extract may further include a flower dimension. The flower dimension material can be used without limitation as long as it is a flower dimension material commonly used for food, and may be, for example, buckwheat whole plant, rose flower, marigold flower, and / or hibiscus flower. The buckwheat vinegar extract (buckwheat vinegar extract blended with the flower dimension material) further comprising the flower dimension material may have improved taste.

Buckwheat ( Fagopyrum spp.) is a food crop belonging to the buckwheat genus and has a short cultivation period of 60-90 days from sowing to harvest and mainly uses seeds. The buckwheat is not usually limited as long as it is commercially available buckwheat, but, for example, general buckwheat (sweet buckwheat, common buckwheat, sweet buckwheat, Fagopyrum esculentum Moench) (e.g., double buckwheat), bitter buckwheat (sweet buckwheat) , bitter buckwheat, tartary buckwheat, Fagopyrum tataricum L. Gaertn) (eg Golden Miso), or a combination thereof, specifically bitter buckwheat. In addition, the buckwheat variety is not limited as long as it is available on the market, but may be, for example, double-breasted buckwheat, golden smile, or a combination thereof, and specifically golden smile.

In one example, the buckwheat may be buckwheat seed or buckwheat sheath, and the present inventors have confirmed that the buckwheat seed vinegar extract has excellent hangover prevention, improvement, and/or relief effects.

In the present specification, "prevention or elimination of hangover" refers to a phenomenon in which various symptoms that appear after drinking are prevented, alleviated, improved, or cured. Specifically, it may mean reducing the concentration of acetaldehyde in the blood and improving symptoms such as headache, nausea, vomiting, dizziness, and muscle pain.

80-90% of alcohol entering the body is first decomposed into acetaldehyde by alcohol dehydrogenase (ADH) present in liver cells, and then metabolized by acetaldehyde dehydrogenase (ALDH) enzyme to acetic acid (acetic acid). ) is formed and then hydrolyzed to carbon dioxide and water to completely decompose. Acetaldehyde produced by ADH is a major cause of liver damage and is a causative substance such as hangovers.

The composition according to one embodiment promotes alcohol and/or acetaldehyde decomposition or

It may be to increase alcohol dehydrogenase (ADH) and/or acetaldehyde dehydrogenase (ALDH) activity. The present inventors found that the composition according to one embodiment has alcohol and/or acetaldehyde decomposition promoting activity; And/or ADH and/or ALDH enzyme activity promotion, etc. was confirmed to be excellent, and it was confirmed that the composition according to one embodiment exhibits excellent hangover preventing, alleviating and/or relieving effects.

A composition according to one embodiment may be a pharmaceutical composition. When the composition according to one embodiment is in the form of a pharmaceutical composition, it may contain a pharmaceutically effective amount of buckwheat vinegar extract, and may further contain a pharmaceutically (or physiologically) acceptable carrier.

At this time, the pharmaceutically acceptable carrier is one commonly used in the formulation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose , polyvinyl pyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil, but are not limited thereto. In addition to the above components, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, and the like may be further included.

A pharmaceutical composition for preventing or relieving a hangover according to one embodiment may be formulated by a method well known in the art to control the release rate of the active ingredient after administration (eg, rapid, sustained or delayed release). there is. The dosage form may be in the form of a tablet, pill, powder, sachet, elixir, suspension, emulsion, solution, syrup, aerosol, soft or hard gelatin capsule, sterile injectable solution, sterile powder, and the like.

The pharmaceutical composition according to one embodiment may be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally or topically applied) according to the desired method, and the dosage is determined according to the patient's condition, body weight, and disease. Depending on the degree, drug form, administration route and time, it can be appropriately selected by those skilled in the art.

The pharmaceutical composition according to one embodiment may be administered in a pharmaceutically effective amount. In the present specification, in the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level is the type of disease, severity, and activity of the drug , sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, factors including concomitantly used drugs, and other factors well known in the medical field. The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered single or multiple times. Considering all of the above factors, it is important to administer an amount that can obtain the maximum effect with the minimum amount without side effects, which can be easily determined by those skilled in the art.

Specifically, the effective amount of the pharmaceutical composition according to one embodiment may vary depending on the patient's age, sex, condition, weight, absorption rate, inactivation rate and excretion rate of the active ingredient in the body, type of disease, and concomitant drugs, generally 1 to 500 mg of the composition according to one example per 1 kg of body weight may be administered daily or every other day, or divided and administered 1 to 3 times a day. However, since it may increase or decrease depending on the route of administration, gender, weight, age, etc., the dosage is not limited to the scope of the present invention in any way.

The composition according to one embodiment may be a health functional food composition or food composition including a hangover reliever.

In the present specification, "health functional food" means that the active ingredient (eg, buckwheat vinegar extract) of the present invention is added to food materials such as beverages, teas, spices, gum, confectionery, tablets, encapsulations, powders, suspensions, etc. It is a food manufactured with, which means that it brings a specific effect on health when ingested, but unlike general drugs, it has the advantage of using food as a raw material and not having side effects that may occur when taking the drug for a long time. The health functional food composition according to one embodiment is very useful because it can be consumed on a daily basis.

The food composition according to one embodiment includes all foods such as beverages, meat, chocolate, food, confectionery, pizza, ramen, other noodles, gum, ice cream, alcoholic beverages, and vitamin complexes.

If the composition according to one embodiment is in the form of a health functional food composition or food composition, it can be prepared as a food with high medical and medical effects processed to efficiently display a bioregulatory function in addition to specific health food and nutritional supply, and useful In order to obtain the effect, it can be prepared in various forms such as tablets, capsules, powders, granules, liquids, and pills.

A health functional food composition or food composition according to an example may include additional components that are commonly used in food compositions to improve odor, taste, and visual properties. For example, it may include vitamins A, C, D, E, B ₁ , B ₂ , B ₆ , B ₁₂ , niacin, biotin, folate, panthotenic acid, and the like. there is. In addition, minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), and copper (Cu) may be included. In addition, amino acids such as lysine, tryptophan, cysteine, and valine may be included. In addition, preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dihydroacetate, etc.), disinfectants (bleaching powder, high bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisole (BHA), butylhydroxytoluene (BHT) ), etc.), coloring agents (tar color, etc.), coloring agents (sodium nitrite, sodium nitrite, etc.), bleaching agents (sodium sulfite, etc.), seasonings (MSG sodium glutamate, etc.), sweeteners (dulcin, cyclemate, saccharin, sodium, etc.), Food additives such as flavoring (vanillin, lactones, etc.), leavening agent (alum, D-potassium hydrogentartrate, etc.), strengthening agent, emulsifier, thickener (thickener), coating agent, gum base agent, foam inhibitor, solvent, improver, etc. can be added. The additive may be selected according to the type of food and used in an appropriate amount.

When using the health functional food composition or food composition according to one embodiment as a food additive, it may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method.

In the health functional food composition or food composition according to one embodiment, the content of the buckwheat vinegar extract is not particularly limited, and may be variously changed depending on the condition of the subject to be administered, the type of specific disease, the degree of progression, and the like. If necessary, it may also be included in the total content of food.

Buckwheat vinegar extract according to one embodiment significantly improves alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activities, and rapidly decomposes alcohol and acetaldehyde in the body to prevent or relieve a hangover. The effect is excellent.

Figure 1 shows the experiment schedule according to the change in alcohol concentration according to the administration of buckwheat vinegar extract, blood acetaldehyde content measurement in an in vivo hangover-induced animal model through alcohol administration.
Figures 2a and 2b shows the results of measuring the change in alcohol concentration according to the buckwheat vinegar extract administration in a hangover-induced animal model through alcohol administration. Specifically, FIG. 2a shows the change in alcohol concentration over time, and FIG. 2b shows the area under the curve (AUC) over time.
Figure 3 shows the results of measuring the acetaldehyde content in the blood according to the buckwheat vinegar extract administration in a hangover-induced animal model through alcohol administration.
Figure 4 shows the results of measuring the ADH activity in the blood and liver tissue of a mouse whose hangover was induced by alcohol administration.
5 shows the results of measuring the ALDH activity in blood and liver tissue of mice whose hangover was induced by alcohol administration.
6 shows the results of quantification of SOD in liver tissue of mice in which hangover was induced by alcohol administration.
Figure 7 shows the quantitative results of Catalase in the liver tissue of a mouse hangover induced by alcohol administration.
In Figures 2 to 7, NOR is a normal group (Normal mice group); CON is a negative control group induced hangover by alcohol administration (Alcohol-induced hangover mice group); SCB is a seed of common buckwheat treated group; And STB represents the seed of tartary buckwheat treated group. The experimental results of FIGS. 2 to 7 are presented as mean ± standard error (n = 5).
8 and 9 show buckwheat vinegar extracts and buckwheat vinegar extracts (buckwheat golden smile vinegar, buckwheat whole plant vinegar, buckwheat marigold vinegar, buckwheat rose vinegar, buckwheat hibiscus vinegar) blended with the buckwheat vinegar extract and the flower dimension according to an example.
Figure 10 shows the buckwheat vinegar extract blended with the flower dimension used for taste evaluation.

The present invention will be described in more detail with the following examples, but the scope of rights is not intended to be limited to the following examples.

Reference Example 1. Preparation of experimental materials and buckwheat vinegar extract

In this experiment, two varieties of normal buckwheat ( Fagopyrum esculentum Moench ), 'Hangjeol buckwheat' and bitter buckwheat ( Fagopyrum tataricum Gaertner ) 'Golden Miso', cultivated by the National Institute of Food Science and Technology of the Rural Development Administration, were used. Seeds sown and harvested in the test field located at 128.45°, 750 m) were used as experimental materials. 2 kg of each sample (buckwheat seed) was measured, immersed in water for 30 minutes, steamed at 100 ° C for 30 minutes, dried in a dryer at 40 ° C for 3 days, and then soaked in 20 L of brown rice vinegar to extract. That is, brown rice vinegar was added 10 times the weight of buckwheat, extracted by immersion at 20 ± 2 ° C for 14 days, and then the precipitate was removed using a filter to prepare buckwheat vinegar extract. In a later experiment, the buckwheat vinegar and distilled water were diluted at a ratio of 1:1 and administered to the rats because the concentration of brown rice vinegar was too thick to administer the undiluted extract.

Reference Example 2. Rutin, quercetin, and choline analysis

Reference Example 2-1. Rutin and quercetin assay

Buckwheat seeds grown in Daegwallyeong-myeon, Pyeongchang-gun, Gangwon-do, buckwheat flowers, leaves, and stems sampled during flowering are harvested and stored in a -70°C ultra-low temperature freezer (deep freezer, Ilsin BioBase Co. Ltd., Yangju, Korea). ) for 24 hours, and then dried for 96 hours using a freeze dryer (Iksan BioBase Co., Ltd., Dongducheon, Korea) (Fig. 1). The lyophilized sample was ground in a grinder (Grinder SFM-555 SP, Shinil Co., SeouL, Korea), sieved through a 40 mesh sieve, and then powdered to be used for later sample analysis (rutin, quercetin, and choline analysis). In addition, buckwheat seeds and vinegar extracts from outpost were used for analysis and compared.

Rutin and quercetin analysis of buckwheat raw materials was analyzed by UPLC according to the method of Kim et al. (2017b). 100 times 100% methanol (J.T. Baker Avantor Korea Co. Suwon, Korea) was added to freeze-dried seed and leaf powder samples, and 80% methanol was added using a Soxhlet heater (Soxhlet heater DH-43, Jisico Sci., SeouL, Korea). Useful components were extracted by reflux extraction for 1 hour in a constant temperature water bath. The extract was filtered through a filter paper (No. 6, Whatman, Maidstone, UK), transferred to a volumetric flask, diluted to 100 mL, and filtered again with a membrane filter (PTFE 13mm 0.20 ㎛, PALL Life Sciences, Ann Arbor, MI, USA) After that, it was analyzed by Ultra Performance Liquid Chromatograph (UPLC). An analytical column (Acquity UPLC CSH C18, 2.1 mm i,d, 100 mm length, 1.7 μm particle size, Waters Corporation, Milford, MA, USA) was created and analyzed. As reagents used in the analysis, HPLC-grade purity acetonitrile (acetonitrile, Tedia Co. Cincinnati, OH, USA) and formic acid (Sigma-Aldrich Co. St. Louis, MO, USA) were used, and distilled water was Distilled water purified in an ultra-pure distilled water maker (Milli-Q system, Millipore, Bedford, MO, USA) was used. As a mobile phase, solvent A (1% formic acid in water, v/v) and solvent B (0.1% formic acid in acetonitrile, v/v) were used and flowed at a flow rate of 0.25 mL/min. Solvent delivery was in a gradient manner, starting with solvent B at a concentration of 7%, increasing to 17% from 2 to 11 minutes, increasing again to 25% from 11 to 13 minutes, and then increasing again to 19 minutes. concentration was maintained. Then, solvent B was reduced from 25% to 7% from 19 to 21 minutes and then stabilized at a concentration of 7% for 2 minutes until 23 minutes. The column in the UPLC instrument was set to a temperature of 30 ° C and the extract was set to a temperature of 20 ° C, the detection wavelength was set to 259 nm, and the sample injection volume was set to 1 μL. The standard materials used in the analysis were rutin (rutin, Extrasynthese, Genay, France) and quercetin (quercetin, Extrasynthese, Genay, France) with a purity of ≥99%, and the calibration equation was analyzed by UPLC for each concentration and the calibration curve was written and calculated.

Reference example 2-2. choline assay

Choline analysis of buckwheat raw materials was analyzed by LC/MS/MS, and the standard material used for analysis was dissolved using 90% Acetonitrile. 400 mg of glass bead and 12 mL of 90% Acetonitrile were added to 400 mg of lyophilized and pulverized seeds of soybean buckwheat and golden miso, respectively, and mixed, followed by sonication for 30 minutes. The sample mixture was repeatedly put in an Ep-tube by 1 mL three times and centrifuged at 10 ° C. at 10,000 rpm for 10 minutes to obtain a supernatant. For final 10-fold dilution, 100 μL of the internal standard diluent (1 ppm) and 800 μL of 90% acetonitrile were added to 100 μL of the sample supernatant and diluted. The mixed sample was filtered through a 0.22 μm PVDF syringe filter (Whatman) and analyzed by putting 500 μL in a 2 mL vial.

An analytical column (Infinity Lab porochell 120 hilic column, 2.1x100mm, 2.7micron) was mounted on an LC (Agilent 1260) instrument equipped with an LC/MS (Agilent 6410 Triple quadrupole LC/MS) and analyzed (Table 1; choline analysis HPLC conditions for).

HPLC-grade purity acetonitrile (acetonitrile, J.T. Baker®, USA) and methanol (methanol, J.T. Baker®, USA) were used as reagents used in the analysis, and distilled water was used in an ultra-pure distilled water maker (Milli-Q system, Millipore, USA). Bedford, MO, USA) purified distilled water was used. Solvent A (10 mM ammonium acetate in DI water, w/v) and solvent B (100% acetonitrile) were used as mobile phases and flowed at a flow rate of 0.3 mL/min. Solvent delivery was carried out in a gradient manner, starting with solvent A at a concentration of 90%, reducing to 50% from 0 min to 5 min, and then to 40% again by 6 min. It was maintained at that concentration from 6 minutes to 8 minutes, increased to 90% from 8 minutes, and maintained until 10 minutes. The column temperature in the LC instrument was set to 30 ° C, and the sample injection amount was set to 1 μL. Standard materials used in the analysis were choline chloride (Sigma-Aldrich Co. St. Louis, MO, USA) with a purity of ≥99%, choline chloride (trimethyl-D9), Sigma-Aldrich Co. St. Louis, MO, USA) was used, and the retention time was 4.219 min. The calibration equation was calculated after preparing a calibration curve by analyzing the standard material by LC for each concentration.

Reference Example 3. Experimental animals and breeding conditions

Experimental animals purchased 5-week-old male SD rats (Hanabio tech, Seongnam, Korea), maintained the growth conditions in the animal laboratory at an appropriate temperature (21 ± 2 ℃ and humidity (50 ± 10%)), and adjusted the light and shade every 12 hours After sufficient adaptation, the experiment schedule was started.The experiment schedule is shown in Fig. 1. Before the test, the weight of all subjects was measured, and the test was conducted after separating the groups so that the average weight of each group was uniform. AIN-93G diet was supplied to the experimental animals, and food and drinking water were freely consumed.Experimental groups were normal group (NOR), alcohol control group (CON), seed of common buckwheat treatment group (SCB), and bitter buckwheat. It was divided into 4 groups (5 animals per group) of Seed of Tartary Buckwheat (STB) (Table 2).

experimental group treatment Normal group (NOR) Plain Vinegar Administration Negative Control (CON) Plain Vinegar + Alcohol Administration Double-cut Buckwheat (SCB) Vinegar containing buckwheat seeds + alcohol administration Bitter Buckwheat (STB) Vinegar containing golden smile seeds + alcohol administration

Ethanol at a concentration of 3 g/kg (body weight) was orally administered to rats at a dose of 10 mL/kg to induce hangover in the other groups except for NOR. 30 minutes before alcohol treatment, buckwheat vinegar extract was orally administered at a dose of 10 mL/kg to the test group, and ordinary vinegar was orally administered at a dose of 10 mL/kg to the normal group and the control group. All animal experiments were conducted after obtaining approval from the SEMI Institutional Animal Care and Use committee of Dongnam Chemical Research Institute (approval number: SEMI-21-008).

Reference Example 4. Quantification of blood acetaldehyde/alcohol

For quantification of acetaldehyde in the blood, the rats were divided into 4 groups (5 animals per group) and general vinegar or buckwheat vinegar extract was orally administered at a dose of 10 mL/kg, and after 30 minutes, 3 groups were orally administered ethanol at a dose of 10 mL/kg. administered. Blood was collected from the tail before ethanol administration and 30, 60, 90, 120, and 180 minutes after administration. The collected blood was left at room temperature for 30 minutes and then centrifuged at 1,700 Х g for 15 minutes to separate serum. Some immediately measured the blood ethanol concentration by hour, and some were stored at -70 ℃ to measure the concentration of acetaldehyde in the future used in Quantification of blood ethanol was measured according to the Cell biolabs protocol using an alcohol assay kit (colorimetric) (STA-620, Cell Biolabs, INC. USA). That is, 87 μL of distilled water, 10 μL of 10X assay buffer, 1 μL of 100X enzyme mixture, and 2 μL of 50X colorimetric probe were mixed with the separated serum (10 μL), incubated at 37°C for 30 minutes in a light-shielded state, and absorbance at 540-570 nm was measured. measured. Quantification of acetaldehyde in blood was measured according to the abcam protocol using an aldehyde quantification assay kit (colorimetric) (ab112113, abcam, USA). That is, after mixing 40 μL of the separated serum (10 μL) diluent buffer and 50 μL of yellow indicator (in assay buffer) and incubating at room temperature for 30-60 minutes, the absorbance was measured at 405 or 550 nm. The concentration of blood alcohol and acetaldehyde was calculated using a calibration curve prepared using a standard for each concentration.

Reference Example 5. Measurement of ADH/ALDH activity

In vitro ADH activity was measured according to the Abnova protocol using an alcohol dehydrogenase assay kit (KA3785, Abnova, Taiwan), and ALDH activity was measured using an ALDH activity assay kit (ab155893, abcam, USA) to measure blood alcohol over time. After confirming the change in concentration, the experimental animals were treated at the time when the change in blood alcohol concentration was the highest, blood was collected from the abdominal aorta, and liver tissue was removed. The collected blood was placed in an sst tube (serum seperate tube), left at room temperature for 30 minutes, centrifuged at 1,700 Х g for 15 minutes, separated from serum, stored at -20 ° C, and used for future ADH and ALDH activity measurements. Liver tissue excised at autopsy was frozen at -20 ° C immediately after weighing, then pulverized in solutions for each analysis item, and the supernatant was separated and analyzed. The liver tissue for ADH analysis was ground in physiological saline at a ratio of tissue to physiological saline at a ratio of 1:4, and then centrifuged at 10,000 x g for 15 minutes (4° C.) to separate the supernatant and used in the experiment. After dispensing the liver enzyme supernatant (20 μL) into each of the two wells, 80 μL of the working reagent (substrate 5 μL, MTT solution 14 μL, NAD solution 9 μL, Diaphorase 1 μL, assay buffer 55 μL) was dispensed into one well. , Dispense 80 μL of blank working reagent (MTT solution 14 μL, NAD solution 9 μL, Diaphorase 1 μL, assay buffer 60 μL) into the remaining wells and mix well. After measuring the absorbance at 565 nm using a microplate reader, it was measured once more after 30 minutes. Liver tissues for ALDH analysis were pulverized using assay buffer, a component of the kit, at a ratio of tissue and assay buffer of 1:4, and then centrifuged at 12,000 rpm for 5 minutes to separate the supernatant and use in the experiment. Add 25 μL of assay buffer to the liver enzyme supernatant (25 μL), dispense 50 μL of the reaction mix, mix well, incubate at room temperature for 5 minutes, and then measure absorbance (450 nm). ALDH activity was evaluated by re-measuring absorbance after 20 to 60 minutes.

Reference Example 6. Quantification of SOD/Catalase

SOD and catalase in the body were quantified using liver tissue obtained from the above procedure. SOD in liver tissue was measured according to the Mybiosource protocol using the Rat Superoxide Dismutases (SOD) ELISA Kit (MBS2707324, Mybiosource, USA), and catalase was measured according to the Mybiosource protocol using the Rat Catalase ELISA kit (MBS726781, Mybiosource, USA). measured. Liver tissues for SOD analysis were ground in a phosphate buffer solution (0.01 mol/L PBS) at a ratio of tissue to PBS of 1:20, centrifuged at 10,000 x g for 5 minutes, and the supernatant was separated and tested. After incubating the liver enzyme supernatant (100 μL) at 37°C for 1 hour, 100 μL of Detection reagent A was added and incubated at 37°C for 1 hour. After washing, 100 μL of Detection reagent B was added, and 90 μL of substrate solution was added to each well after additional incubation and washing at 37 ° C for 30 minutes. After 10-20 minutes of incubation at 37°C in a light-shielded state, 50 μL of stop solution was added, and absorbance was measured at 450 nm after mixing. Liver tissues for catalase analysis were ground in a phosphate buffer solution (0.02 mol/L PBS) at a ratio of tissue and PBS of 1:1, and then centrifuged at 1,500 x g for 15 minutes to separate the supernatant before testing. After adding 10 μL of Balance solution to 100 μL of supernatant, 50 μL of congugate was added, mixed well, and incubated at 37 ° C for 1 hour. After washing, 50 μL each of Substrate A and B was added, and after incubation at 37° C. for 15-20 minutes, 50 μL of stop solution was added, mixed, and absorbance was measured at 450 nm.

Reference Example 7. Statistical Analysis

All test results were expressed as mean and standard deviation (mean±SD), and statistical analysis for each result was performed by t-test One-way ANOVA using SAS or Statview program, and the statistical significance of the average value was p -value was tested with

Example 1. Comparison of rutin / quercetin / choline content of normal buckwheat and bitter buckwheat

The rutin and quercetin contents of the raw materials and vinegar extract of the seeds and outposts of normal buckwheat and bitter buckwheat were measured in the same manner as in Reference Example 2-1, and the results are shown in Table 3 below. As shown in Table 3, the rutin content was 74.7 mg / 100 g in the bitter buckwheat seed extract and 14.7 mg / 100 g in the seed, respectively, higher than that of the general buckwheat seed. Quercetin component was detected in a relatively large amount (6.9-7.1 mg/100 g) in the outposts of general buckwheat and bitter buckwheat, and was high in bitter buckwheat. Bitter buckwheat has a higher rutin content than normal buckwheat, so various studies are being conducted to use it as a food and medicine material (Yoon et al., 2006; Yoon et al., 2012). It was found to be higher than that of vinegar extract, showing a tendency consistent with the results of previous studies.

Material ¹⁾ Rutin (mg/100g) Quercetin (mg/100g DW) SCB 33.1±0.4 ²⁾ 0.0±0.00 STB 1,459.1±7.3 0.0±0.00 PCB 3,171.5±6.3 15.5±1.4 PTB 4,470.5±6.1 36.8±0.7 SCBV 13.5±0.1 ²⁾ 0.8±0.03 STBV 14.7±0.1 0.0±0.00 PCBV 60.1±0.3 6.9±0.24 PTBV 74.7±0.3 7.1±0.05

¹⁾ Ingredients were harvested in Pyeongchang. SCB, common buckwheat seed; STB, bitter buckwheat seed; PCB, common buckwheat outpost; PTB, bitter buckwheat outpost; SCBV, common buckwheat seed-containing vinegar extract; STBV, vinegar extract containing bitter buckwheat seeds; PCBV, ordinary buckwheat whole grain vinegar extract; Vinegar extract containing PTBV, bitter buckwheat sprouts

²⁾ Data ± standard deviation

The choline content of vinegar extract, seeds and outposts of normal buckwheat and bitter buckwheat was measured in the same manner as in Reference Example 2-2, and the results are shown in Table 4 below. Choline plays an important role in maintaining liver function, general brain development, neurological function, and healthy metabolism, and exists in the form of phospholipid phosphatidylcholine. Choline is a water-soluble nutrient related to folic acid and vitamins. Also, like B vitamins, choline plays an important role in maintaining metabolic activity as well as energy support and brain function. Choline is involved in detoxification, nerve signaling and the function of an important neurotransmitter called acetylcholine, which acts as an anti-aging neurotransmitter and helps nerve cells communicate with each other while moving muscles.

As shown in Table 4, common buckwheat vinegar extract and bitter buckwheat vinegar extract had high choline content .

Material ¹⁾ Choline (ug/g) Choline-d9 (Recovery, %) SCBV 1296.21 103.8 - 129.3 STBV 1226.52 94.9 - 103.2

¹⁾ Ingredients were harvested in Pyeongchang. SCBV, common buckwheat seed-containing vinegar extract; STBV, vinegar extract containing bitter buckwheat seeds

Example 2. Quantification of Acetaldehyde/Alcohol in Blood

As in the method of Reference Example 4, changes in blood alcohol and acetaldehyde concentrations over time were measured in each experimental group, and the results are shown in FIGS. 2A to 3 . The change in blood alcohol concentration over time was graphed to calculate the area under the curve (AUC) over time (FIG. 2b).

As shown in FIG. 2a, the time for the blood alcohol level to reach its peak was 60 minutes in all groups, and the blood alcohol level 60 minutes after alcohol administration was the highest in the CON group at 2006.0 μmole. In the SCB group, it was 1754.4 μmole, which was 12.5% lower than that of the CON group, and in the STB group, it was 1674.7 μmole, which was 16.5% lower than that of the CON group. In addition, as shown in FIG. 2B, when comparing AUC values of changes in blood alcohol concentration, there was a tendency to decrease by 11.5% in the SCB group (186,984.8) and by 36% in the STB group (135,183.0) than in the CON group (211,360.0). No level difference was seen.

As shown in FIG. 3, the concentration of acetaldehyde in blood over time continuously increased over time in the CON group, but in the SCB group, after reaching the highest concentration at 120 minutes (13.57 μmole), it decreased at 180 minutes (12.16 μmole), and in the STB group In , after reaching the highest concentration at 90 minutes (14.14 µmole), it gradually decreased at 120 minutes (13.57 µmole) and 180 minutes (10.63 µmole).

Therefore, both the general buckwheat seed vinegar extract and the bitter buckwheat seed vinegar extract were effective in reducing the change in blood alcohol concentration and the change in acetaldehyde concentration, and in particular, it was confirmed that the bitter buckwheat seed vinegar extract exhibited an excellent effect.

Example 3. Measurement of ADH/ALDH activity in blood

As in the method of Reference Example 5, in vivo ADH and ALDH activities were measured by treating the experimental animals 60 minutes after alcohol administration, obtaining liver tissue and blood, and then measuring them by ELISA assay, and the results are shown in FIGS. 4 and 5, respectively. 5.

As shown in FIG. 4, the concentration of ADH in the blood decreased to 1.708 U/L in the CON group (alcohol control group) compared to the SCB group (2.384 U/L) and the STB group (2.443 U/L), confirming that the alcohol decomposition ability was lowered. could ADH concentrations in the SCB and STB groups increased by 40% and 43%, respectively, compared to the CON group. The ADH enzyme concentration in liver tissue was 0.722 U/L in the CON group, which was lower than that of the SCB group (1.891 U/L) and the STB group (2.099 U/L), confirming that the alcohol decomposition ability was lowered. In the SCB and STB groups, the ADH concentration increased by 162% and 191%, respectively, compared to the CON group. The NOR group showed a significantly higher ADH concentration than the CON group, but there was no significant difference in the sample group.

As shown in Figure 5, the concentration of ALDH enzyme also showed a similar trend to the ADH concentration. Blood ALDH concentration increased by 19% in the SCB group (3.144 mU/mL) and by 21% in the STB group (3.194 mU/mL), compared to the CON group (2.641 mU/mL). In liver tissue, it increased by 41% in the SCB group (1.693 mU/mL) and by 47% in the STB group (1.768 mU/mL) compared to the CON group (1.200 mU/mL). ALDH concentration did not show a significant difference in the whole group.

Based on these results, it is thought that common buckwheat seeds and bitter buckwheat seeds increase the amount of alcohol decomposing enzyme in liver tissue and blood, promoting the decomposition of acetaldehyde, the main cause of hangover.

Example 4. Quantification of Superoxide Dismutases/Catalase

As in the method of Reference Example 6, after treating the control group and each experimental group, the liver tissues extracted were pretreated to quantify SOD and Catalase in the body, and the results are shown in FIGS. 6 and 7, respectively.

As shown in FIG. 6, the SOD concentration in liver tissue was significantly increased to 2.972 ng/mL (#p<0.005) in the STB group compared to the CON group (1.617 ng/mL), and to 2.285 ng/mL in the SCB group ( *p<0.05) significantly increased. Among them, the STB group administered with bitter buckwheat seed vinegar extract showed a 23% higher SOD concentration than the SCB group administered common buckwheat seed vinegar extract.

As shown in Figure 7, the concentration of catalase also increased significantly (+p<0.0001) to 17.010 ng/mL in the STB group compared to the CON group (7.953 ng/mL), and to 10.672 ng/mL in the SCB group (*p< 0.05) increased significantly. Among them, catalase concentration was 37% higher in the STB group than in the SCB group.

Antioxidant defense mechanisms and antioxidant enzymes such as catalase, superoxide dismutase, and glutathione peroxidase of oxidation mechanisms are important enzymes that detoxify free radicals and have been reported to be closely related to life extension. In the antioxidant enzyme activity test that measures liver protection function, SOD (superoxide dismutase) and catalase enzyme activities were higher in buckwheat vinegar extract than in the existing negative control group, respectively, and in the liver damage recovery test, AST and ALT, which are values indicating liver damage recovered more than the negative control group.

Example 5. Blending of Buckwheat Vinegar and Flower Dimensions

Same as or similar to the method of Reference Example 1, brown rice vinegar was added 10-30 times the weight of the raw grain at 20 ± 5 ° C and extracted (or maintained) for about 2-3 weeks to ferment buckwheat vinegar (buckwheat vinegar extract) ) was prepared,

After adding 10-30 times brown rice vinegar sugar compared to the weight of raw grains, buckwheat starch, rose flower, marigold flower or hibiscus flower were blended at 2-3% each and extracted (or maintained) for about 2-3 weeks, Buckwheat vinegar extracts (buckwheat golden misocho, buckwheat whole herb vinegar, buckwheat marigold vinegar, buckwheat rose vinegar, buckwheat hibiscus vinegar) blended with floral materials were prepared.

The buckwheat vinegar extract prepared above and the buckwheat vinegar extract (buckwheat golden misocho, buckwheat whole herb vinegar, buckwheat marigold vinegar, buckwheat rose vinegar, buckwheat hibiscus vinegar) blended with the floral materials are shown in FIGS. 8 and 9, respectively. .

Appetite evaluation was performed on the buckwheat vinegar extract prepared above and the buckwheat vinegar extract blended with the floral material, and the results are shown in Table 5 and FIG. 10. As shown in Table 5 and FIG. 10, various tastes could be felt.

division Appetite evaluation ¹⁾ Sour taste sweetness Scent clean taste Umami rain forest comprehensive evaluation Buckwheat Golden Misocho 4 4 3 4 4 3 clean and comfortable taste Buckwheat Whole Vinegar One One One One One 4 It has a strong smell and a beautiful color buckwheat marigold vinegar 4 4 2 3 3 2 Strong buckwheat gold flavor buckwheat rose vinegar 5 5 5 5 5 4 Pleasant refreshing taste buckwheat hibiscus 2 4 4 2 3 5 Slightly bitter taste and nice color

¹⁾ Appetite evaluation score: 1, very poor; 2, insufficient; 3, normal; 4, excellent; 5, very good

Claims (17)

A pharmaceutical composition for preventing or relieving a hangover, comprising a buckwheat vinegar extract.
The pharmaceutical composition according to claim 1, wherein the buckwheat vinegar extract is obtained by adding 15 to 25 times more vinegar than buckwheat by weight.
The pharmaceutical composition according to claim 1, wherein the buckwheat is a buckwheat seed or buckwheat sheath.
According to claim 1, wherein the buckwheat is common buckwheat, bitter buckwheat, or a combination thereof, the pharmaceutical composition.
The pharmaceutical composition according to claim 1, wherein the buckwheat is double-breasted buckwheat, golden smile, or a combination thereof.
The pharmaceutical composition according to claim 1, wherein the composition promotes decomposition of alcohol or acetaldehyde.
The pharmaceutical composition for preventing or relieving hangover according to claim 1, wherein the composition increases the activity of alcohol dehydrogenase (ADH) or acetaldehyde dehydrogenase (ALDH).
The pharmaceutical composition according to claim 1, wherein the buckwheat vinegar extract further comprises a flower tea raw material.
Health functional food composition for preventing or relieving hangover, containing buckwheat vinegar extract.
10. The method of claim 9, wherein the buckwheat vinegar extract is obtained by adding 15 to 25 times the vinegar compared to buckwheat by weight, health functional food composition.
10. The method of claim 9, wherein the buckwheat is a buckwheat seed or buckwheat sheath, health functional food composition.
10. The method of claim 9, wherein the buckwheat is normal buckwheat, bitter buckwheat, or a combination thereof, health functional food composition.
10. The method of claim 9, wherein the buckwheat is yangjeol buckwheat, golden smile, or a combination thereof, health functional food composition.
10. The method of claim 9, wherein the composition promotes the decomposition of alcohol or acetaldehyde, health functional food composition.
10. The method of claim 9, wherein the composition is alcohol dehydrogenase (alcohol dehydrogenase; ADH) or acetaldehyde dehydrogenase (aldehyde dehydrogenase; ALDH) to increase the activity, health functional food composition.
10. The method of claim 9, wherein the buckwheat vinegar extract is a health functional food composition further comprising a flower tea raw material.
A food composition for preventing or relieving a hangover, comprising a buckwheat vinegar extract.

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