KR102413920B1 - Hangover-relieving composition comprising fermented black radish - Google Patents

Abstract

The present invention relates to a composition for relieving a hangover comprising a fermented black-free lactic acid bacteria.

Description

Hangover-relieving composition comprising fermented black radish

The present invention relates to a composition for relieving a hangover comprising a fermented black-free lactic acid bacteria.

Excessive alcohol intake affects almost all parts of the body, causing many diseases such as liver disease, gastritis, pancreatitis, high blood pressure, stroke, diabetes, heart disease, thirst, general malaise, fatigue, memory loss, abdominal bloating, and digestion. It causes many side effects, including hangover symptoms such as malnutrition, vomiting, diarrhea, vitamin deficiency, and the risk of alcohol poisoning.

Among them, hangover phenomenon is known to be caused by the action of alcohol and acetaldehyde accumulated in the body. Ethanol entering the liver is converted to acetaldehyde by alcohol dehydrogenase (ADH) in the cell cytosol, and then converted to acetate by the action of aldehyde dehydrogenase (ALDH), which is converted into hepatocytes through the circulatory system. excreted out Acetaldehyde, which is produced as an initial metabolite of ethanol in the above metabolic process, is known to be highly reactive and highly toxic than ethanol. In particular, acetaldehyde is a representative chemical that accompanies a headache after drinking, and the WHO has warned of its toxicity.

In a normally functioning human body, ALDH, the main enzyme involved in the oxidation of aldehydes, is widely distributed in organs and tissues including the liver, kidneys, heart and stomach, so it can respond to the side effects of various aldehydes in the body. However, if ALDH activity is changed by a person who is congenitally deficient in ALDH or due to factors such as excessive drinking, normal metabolism is disturbed by the toxicity of unoxidized ethanol and active aldehyde, making it impossible to respond to hangovers and various side effects. do.

Therefore, there are various developments of products containing herbal or artificial preparations capable of removing a hangover caused by alcohol intake and reducing blood alcohol and acetaldehyde concentrations. A composition for relieving hangover and antioxidant action containing the extract is disclosed.

Black radish is a kind of radish, a root vegetable of the cruciferous family, also called black radish or black radish, and the scientific name is Raphanus sativus L. var. known as niger . The outer surface is black, but the inner white flesh is composed of white flesh, and it is known to exhibit various physiological activities such as anti-hyperlipidemia, anti-inflammatory, antioxidant, and toxin removal. In the present invention, it was found that the components contained in the black radish exhibit excellent effects on the control of blood alcohol concentration, ADH and ALDH levels, and thus, a composition for relieving a hangover was developed.

The present invention provides a composition for relieving a hangover having an excellent alcohol decomposition effect.

The composition for relieving a hangover of the present invention includes a black-free lactic acid bacteria fermented product.

The composition for relieving a hangover according to the present invention has excellent alcohol decomposition effect and exhibits an excellent hangover relieving effect by promoting the activity of ADH and ALDH.

1 is a graph showing the ADH activity according to the concentration of the composition of the present example.
2 is a graph showing ALDH activity according to the concentration of the composition of the present example.
3 is a graph showing the blood ethanol concentration of the animals administered the composition of the present example.
4 is a graph showing the concentration of acetaldehyde in the blood of animals administered the composition of the present example.
5 is a graph showing the ADH activity of the liver tissue of the animal administered the composition of the present example.
6 is a graph showing the ALDH activity of the liver tissue of the animal administered the composition of the present example.
7 is an image of H&E-stained slides of liver tissues of animals administered with the composition of the present embodiment.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated. However, it is not limited only by the following content, and each component may be variously modified or selectively mixed as needed. Accordingly, it should be understood to include all modifications, equivalents and substitutes included in the spirit and scope of the present invention.

The hangover relieving composition of the present invention utilizes black radish, and in particular, includes black radish fermented lactic acid bacteria. When black radish is fermented with lactic acid bacteria, the effect of lowering the blood alcohol concentration and promoting the activities of ADH (Alcohol dehydrogenase) and ALDH (Acetaldehyde dehydrogenase) becomes more excellent, and at the same time, it is easy to consume.

For example, the fermented black radish lactic acid bacteria according to the present invention includes a fermented black radish of Lactobacillus spp. The Lactobacillus genus lactic acid bacteria is Lactobacillus plantarum ( Lactobacillus plantarum ), Lactobacillus casei ( Lactobacillus casei ), Lactobacillus helveticus ( Lactobacillus helveticus ) and at least one selected from the group consisting of Lactobacillus reuteri ( Lactobacillus reuteri ) can For example, the Lactobacillus genus lactic acid bacteria may be Lactobacillus plantarum ( Lactobacillus plantarum ). Lactobacillus plantarum ( Lactobacillus plantarum ) In addition to the effect related to alcohol decomposition, the antioxidant effect is also excellent, so that the synergistic effect on alcohol decomposition can be more excellently exhibited.

The dosage of the assay-free lactic acid bacteria fermented product according to the present invention may be 720 to 2,880 mg/day based on an adult (average body weight of 60 kg). When the assay-free lactic acid bacteria fermented product of the present invention is administered in the above-mentioned range, it is possible to obtain the effect of reducing the blood alcohol concentration within a short time.

As an example, the composition for relieving a hangover of the present invention may include 0.001 to 10% by weight of the black-free lactic acid bacteria fermented product based on the total weight of the composition.

The composition for relieving a hangover of the present invention may be a pharmaceutical composition or a food composition. For example, it may be provided in the form of health foods, such as beverages, pills, jellies, effervescent tablets, etc., containing the hangover relieving composition of the present invention, but is not limited thereto.

The method for producing the fermented black radish lactic acid bacteria according to the present invention is not particularly limited, and includes a grinding process of pulverizing black radish, a fermentation process of inoculating and fermenting the pulverized black radish with lactic acid bacteria, and sterilizing and then pulverizing the fermented black radish. It may be prepared by a method including a powdering process.

In the pulverization process, the washed black radish may be sterilized and then pulverized. Sterilization treatment may be performed at a temperature of 70 to 120 °C, for example 80 to 110 °C, in another example 90 to 100 °C, and the heat treatment may be performed for 30 minutes to 90 minutes. In the case of heat treatment at a temperature lower than the above temperature, bacteria may not be completely removed, and in the case of heat treatment at a temperature higher than the above temperature, the active ingredient for alcohol decomposition of the black radish may escape from the black radish.

The grinding process may further include a sterilization process in which purified water is mixed with the crushed black radish and sterilized.

In the sterilization process, purified water may be added to the pulverized black radish in a weight ratio of 1: 0.5 to 3, for example, 1: 0.7 to 2, in another example, 1: 0.85 to 1.5 by weight. If purified water is added in excess than the above ratio, the amount of raw material input per 1 LOT may decrease in the fermentation process proceeding after the sterilization process, so that the fermentation process may not be performed smoothly, and purified water is added in a small amount than the ratio In this case, the liquidity may be insufficient in the subsequent fermentation process, so that the fermentation process may not be performed smoothly.

The sterilization process may be autoclaving at a temperature of 50 to 150 ℃, for example, 70 to 140 ℃, in another example 90 to 130 ℃. In the case of heat treatment at a temperature lower than the above temperature, the sterilization effect may be reduced, and in the case of heat treatment at a temperature higher than the above temperature, the black radish tissue may be damaged, making it unsuitable for fermentation by lactic acid bacteria. The autoclaving may be performed, for example, for 5 to 60 minutes.

In the fermentation process, sterilized ground black radish Lactobacillus genus lactic acid bacteria 4.4 x 10 ² to 4.4 x 10 ⁹ CFU / 100g, for example, 4.4 x 10 ⁵ to 4.4 x 10 ⁹ CFU / 100g, another example 4.4 x 10 ⁶ to 4.4 x 10 ⁷ CFU/100g can be inoculated. When inoculated less than the above amount, it is not sufficiently fermented, so it is difficult to prepare a fermented product having a significant level of blood alcohol concentration lowering effect and ADH, ALDH activity promoting effect compared to black radish or black radish extract, In case of excessive inoculation, the effect of promoting ADH and ALDH activity may be reduced.

The fermentation may be carried out at a temperature of 20 to 50 °C, for example, 25 to 45 °C, in another example 32 to 40 °C. Fermentation may not occur at a temperature outside the above-mentioned range.

The fermentation may be performed for 12 to 120 hours, for example, 24 to 100 hours, in another example 30 to 72 hours. In the case of fermentation for a shorter time than the above time, the antioxidant active ingredient may not be sufficiently formed, and the lactic acid bacterium fermented product does not show a significant level of blood alcohol concentration lowering effect and ADH and ALDH activity promoting effect compared to black radish or black radish extract. can On the other hand, in the case of fermenting for a longer time than the above time, the active ingredient and the black radish component are decomposed by overfermentation, and the blood alcohol concentration lowering effect and the ADH and ALDH activity promoting effect may be reduced.

The strain used in the present invention may be sold, for example, from the Korea Microorganism Conservation Center (KCCM), but is not limited to the strain received from the institution.

Hereinafter, the present invention will be described in more detail through examples. However, the following examples are provided to help the understanding of the present invention, and the scope of the present invention is not limited to the examples in any sense.

[Production Example: Production of black-free lactic acid bacteria fermented product]

1) Sales of lactic acid bacteria

Lactobacillus plantarum (KCCM 12116) was purchased from the Korea Culture Center of Microorganisms (KCCM) and used for experiments.

2) Seed culture

The received strains were subcultured in a solid medium and a liquid medium and used in the experiment. Specifically, the received strain was inoculated into a solid medium and cultured at 37 ° C. in an incubator for 24 hours. When colonies were formed, inoculated into a liquid medium and cultured at 37 ° C. for 24 hours in an incubator to prepare a seed culture medium. MRS medium (Difco, USA) was used for seed culture.

3) Preparation of black-free lactic acid bacteria fermented product

The black radish was washed with purified water, sterilized by heat treatment at 95° C. for 30 minutes, and then pulverized using a mixer. The pulverized black radish and purified water were mixed in a weight ratio of 1:2, autoclaved at 121° C. for 30 minutes, and then cooled to room temperature.

The seed culture solution cultured in 2) was inoculated with 1% weight of the sterilized ground black radish mixture, and then cultured with shaking in an incubator at 37° C. for 48 hours for liquid fermentation. After sterilization at 95 °C for 30 minutes after completion of fermentation, the fermented product was lyophilized and powdered.

[Experimental Example 1: ADH activity analysis]

After adding the fermented lactic acid bacteria assay of Preparation Example to purified water to prepare a stock having a concentration according to Table 1 below, it was diluted with purified water to prepare a final sample (final). Alcohol dehydrogenase (ADH) activity was analyzed by measuring the absorbance of NADH formed at 450 nm of each sample using Alcohol Dehydrogenase Activity Assay Kit (sigma) and a spectrophotometer. The maximum absorbance at the end of the reaction of each experimental group was expressed as a ratio to the maximum absorbance of the control group, and was calculated by the following formula. Absorbance was measured three times for each sample, and the average values of the calculated ADH activities are shown in Table 1 and FIG. 1 below.

ADH activity = (B/A) Х100

In the above formula,

A is the maximum absorbance of the control (without fermented black radish),

B is the maximum absorbance of the experimental group (each sample).

As shown in Table 1 and FIG. 1, it was confirmed that the activity of ADH was promoted in the experimental group compared to the control group. In addition, it can be confirmed that the ADH activity promoting effect increases as the concentration of the fermented black radish increases.

[Experimental Example 2: ALDH activity analysis]

After adding the fermented lactic acid bacteria assay of Preparation Example to purified water to prepare a stock having a concentration according to Table 2 below, it was diluted with purified water to prepare a final sample (final). Aldehyde Dehydrogenase Activity Using a Colorimetric Assay Kit (sigma) and a spectrophotometer, the absorbance of NADH formed at 450 nm of each sample was measured to analyze Acetaldehyde dehydrogenase (ALDH) activity. The maximum absorbance at the end of the reaction of each experimental group was expressed as a ratio to the maximum absorbance of the control group, and was calculated by the following formula. Absorbance was measured three times for each sample, and the average values of the calculated ALDH activities are shown in Table 2 and FIG. 2 below.

ALDH activity = (B/A) Х100

In the above formula,

A is the maximum absorbance of the control (without fermented black radish),

B is the maximum absorbance of the experimental group (each sample).

As shown in Table 2 and FIG. 2, it was confirmed that the activity of ALDH was promoted in the experimental group compared to the control group. In addition, it can be confirmed that the ALDH activity promoting effect increases as the concentration of the fermented black radish increases.

[Experimental Example 3: Treatment of experimental animals]

The experiment was conducted using SD rats (Sprague-Dawley rat, male, 8 weeks old) purchased from Orient Bio (Sungnam, Korea). The environment of the breeding room was maintained at an average temperature of 22±1℃ and humidity of 50±10%, and the light and dark was controlled at 12-hour intervals, and feed and water were freely supplied for one week during the acclimatization period.

The experimental animals were divided into a control group, a low-dose group and a high-dose group (24 animals in each group), and were fasted for 18 hours before ethanol administration. 30 minutes before alcohol administration, saline 2 ml/kg for the control group, 75 mg/kg of the Lactobacillus plantarum test radish fermented product of the Preparation Example in the low-dose group, and 300 mg of the Lactobacillus plantarum test radish fermented product of the Preparation Example in the high-dose group. /kg was orally administered, and after 30 minutes, 25% ethanol (Merck, Darmstadt, Germany) was orally administered to all animals at a dose of 4 g/kg.

After 1, 2, and 4 hours of ethanol administration, the animal is anesthetized using isoflurane (Isoflurane, Virbac, Carros, France), the abdominal cavity is opened to collect whole blood from the abdominal vena cava , and the remainder was fixed in 10% neutral formalin. The collected whole blood was centrifuged at 3,000 rpm and 4°C for 15 minutes, and serum was separated and stored at -70°C.

[Experimental Example 4: Measurement of ethanol and acetaldehyde content in blood]

Ethanol and acetaldehyde contents were measured in the serum collected by time. Ethanol was measured with a blood biochemical analyzer (TBA-120FR, Toshiba, Tokyo, Japan), and acetaldehyde was reacted using an Acetaldehyde assay kit (EACT-100, Biovision, Milpitas, CA, USA), followed by a spectrophotometer (Synergy). H4, Biotek, Winooski, VT, USA) was used to measure at 565 nm. Measured values were expressed as mean ± standard deviation. Student's t-test was used to test for statistical significance between the control group and the experimental group, and it was determined to be significant when the P value was less than 0.05.

As shown in FIG. 3 , as a result of analyzing the blood ethanol concentration, the control group showed the highest level of 113.27 mg/dl 1 hour after alcohol administration, 59.00 mg/dl after 2 hours, and 23.85 mg/dl after 4 hours. decreased. In the low concentration group, 1 hour after alcohol administration, it was significantly reduced to 50.07 mg/dl compared to the control group, and after 2 hours it was significantly decreased to 7.37 mg/dl compared to the control group, and after 4 hours, the concentration was 11.22 mg/dl. In the high concentration group, the concentration was significantly reduced to 61.32 mg/dl after 1 hour of alcohol administration, compared to the control group, showed a concentration of 59.30 mg/dl after 2 hours, and decreased significantly to 5.60 mg/dl after 4 hours compared to the control group.

As shown in FIG. 4 , as a result of analyzing the blood acetaldehyde concentration, the control group showed a concentration of 155.05 uM 1 hour after alcohol administration, 169.58 uM after 2 hours, and 131.31 uM after 4 hours. In the low concentration group, the concentration of alcohol was 155.58 uM after 1 hour, 135.54 uM after 2 hours, and 122.93 uM after 4 hours. In the high concentration group, 179.65 uM after 1 hour of alcohol administration, 94.36 uM after 2 hours, and 48.27 uM after 4 hours were significantly reduced compared to the control group.

[Experimental Example 5: Measurement of ADH and ALDH Activity in Liver Tissue]

After autopsy, liver tissue stored at -70°C was divided into 50 mg each to measure ADH and ALDH activity. After reacting using ADH activity colorimetric assay kit (K787-100, Biovision, Milpitas, CA, USA) and ALDH activity colorimetric assay kit (K787-100, Biovision, Milpitas, CA, USA), respectively, spectrophotometer (Synergy H4) , Biotek, Winooski, VT, USA) was used to measure the activity of each enzyme at 450 nm. Measured values were expressed as mean ± standard deviation. Student's t-test was used to test for statistical significance between the control group and the experimental group, and it was determined to be significant when the P value was less than 0.05.

As shown in FIG. 5 , as a result of measuring the ADH activity of liver tissue, it was 108.43 mU/ml in the control group, 123.09 mU/ml in the low concentration group, and 204.35 mU/ml in the high concentration group, which was significantly increased compared to the control group. As shown in FIG. 6 , as a result of measuring the ALDH activity of liver tissue, it was significantly increased compared to the control group to 8.55 mU/ml in the control group, 13.23 mU/ml in the low concentration group, and 19.42 mU/ml in the high concentration group.

From the results of FIGS. 3 to 6 , it was confirmed that the fermented black radish extract according to the present invention increased the activity of ADH and ALDH, thereby affecting the decomposition of ethanol and acetaldehyde after ethanol absorption.

[Experimental Example 6: Histopathological evaluation of liver tissue]

Liver tissue fixed in 10% neutral formalin was dehydrated, cleared, and paraffin-infiltrated using a tissue processor (Excelsior ES, ThermoFisher Scientific, Waltham, MA, USA), followed by tissue embedding (Histostar, ThermoFisher Scientific, Waltham, MA, USA) was used to prepare paraffin blocks. After the paraffin block was cut into 4 µm slices, slides were prepared, and hematoxylin eosin (H&E) staining was performed using an automatic staining machine (Coverstainer, Aglient, Santa Clara, CA, USA). Using a slide scanner (Panoramic SCAN, 3DHistech, Budapest, Hungary), images of H&E-stained slides were taken.

When the liver is damaged by foreign substances, degeneration of hepatocytes, lipidosis, fibrosis, and, in severe cases, necrosis proceeds, and in this case, histopathological observation is possible. . As shown in FIG. 7 , as a result of histopathological evaluation of liver tissue, both the low concentration group and the high concentration group did not show specific changes compared to the control group, and from the above results, the assay-free lactic acid bacteria fermented according to the present invention did not cause toxicological changes. confirmed that it is not.

[Experimental Example 7: Serum Biochemical Analysis]

16 items (Sodium, Potassium, Chloride, Total protein, Albumin, Blood urea nitrogen, Creatinine, Glucose, Total bilirubin, Calcium, Phosphorus, Total cholesterol, Triglyceride, Aspartate transaminase, Alanine aminotransferase and alkaline phosphatase) were measured with a blood biochemical analyzer (TBA-120FR, Toshiba, Tokyo, Japan). Measured values were expressed as mean ± standard deviation. Student's t-test was used to test for statistical significance between the control group and the experimental group, and it was determined to be significant when the P value was less than 0.05.

When the liver is damaged by foreign substances, degeneration of hepatocytes, lipidosis, fibrosis, and, in severe cases, necrosis proceed. Therefore, the concentration of aspartate transaminase and alanine aminotransferase in the blood is an indicator of liver damage. As shown in Table 3 above, in the serum biochemical analysis of the low concentration group and the high concentration group compared to the control group, there was no significant change in all measurement items as well as liver-related aspartate transaminase and alanine aminotransferase. From the above results, it was confirmed that the fermented extract of the present invention does not cause toxicological changes.

Claims (10)

A composition for relieving a hangover comprising a black-free lactic acid bacteria fermented product,
Based on the total weight of the composition for relieving a hangover, a composition for relieving a hangover comprising 0.001 to 10% by weight of the black-free lactic acid bacteria fermented product. The composition for relieving a hangover according to claim 1, wherein the lactic acid bacteria comprises Lactobacillus spp. According to claim 2, wherein the Lactobacillus genus lactic acid bacteria Lactobacillus plantarum ( Lactobacillus plantarum ), Lactobacillus casei ( Lactobacillus casei ), Lactobacillus helveticus ( Lactobacillus helveticus ) and Lactobacillus reuteri consisting of reuteri ( Lactobacillus reuteri ) A composition for relieving one or more hangovers selected from the group. The composition for relieving a hangover according to claim 1, which lowers blood alcohol concentration and promotes the activity of ADH and ALDH. The composition for relieving a hangover according to claim 1, wherein the dosage of the black-free lactic acid bacteria fermented product is 720 to 2,880 mg/day based on an adult (average body weight of 60 kg). delete A pharmaceutical composition comprising the composition for relieving a hangover according to any one of claims 1 to 5. A health food comprising the composition for relieving a hangover according to any one of claims 1 to 5. The health food according to claim 8, which is a beverage, pill, jelly or effervescent tablet. grinding process of grinding black radish;
A fermentation process of inoculating and fermenting the pulverized black radish with Lactobacillus genus lactic acid bacteria; and
As a method for preparing a composition for relieving a hangover comprising a fermented product of black radish lactic acid bacteria comprising a powdering process of sterilizing the fermented black radish and then powdering,
Based on the total weight of the composition for relieving a hangover, the method for preparing a composition for relieving a hangover comprising 0.001 to 10% by weight of the black-free lactic acid bacteria fermented product.

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