KR101371143B1 - Composition comprising chlorella for improving liver function or relieving hangover - Google Patents

Abstract

The present invention relates to a liver function improvement or hangover relief composition comprising chlorella as an active ingredient. The composition for improving liver function or releasing hangover according to the present invention has an effect of reducing the blood alcohol concentration and inhibiting acidification of blood (effect of reducing the concentration of carbon dioxide and lactic acid in the blood, acidified blood pH due to alcohol intake to normal alkali). Conversion effect) is very excellent, and hangover relief effect such as protection from liver damage caused by excessive intake of alcohol, gastrointestinal function improvement effect, headache reduction effect is very excellent.

Description

Composition comprising chlorella for improving liver function or relieving hangover}

The present invention relates to a liver function improving or hangover composition comprising chlorella as an active ingredient and to a liver functional improvement or hangover relief containing chlorella as an active ingredient.

Korea's alcohol consumption is among the highest in the world, and the socioeconomic and health consequences of drinking alcohol are enormous and increasing, with deaths from liver cancer and cirrhosis becoming the leading cause of death for Koreans.

In general, the effect of alcohol absorbed through drinking begins to appear within about 10 minutes after drinking and reaches a peak in 40 to 60 minutes. Alcohol is absorbed in small amounts in the esophagus and oral mucosa, after which about 10% of the alcohol is absorbed in the stomach and the remaining 90% of the alcohol is absorbed in the small intestine. The absorption of alcohol is determined by several factors. Fatty foods slow down the absorption of alcohol because of their slow moving speed. They are absorbed three to four times faster than when drinking with food on an empty stomach, and drink alcohol quickly. The more absorbed, the higher the rate. Alcohol is a substance that produces a high calorie of 7.1 kcal / g from a nutritional point of view, but the body does not absorb the calories it contains. It can also lead to malnutrition because it can not be used as a useful energy source in vivo.

Excessive alcohol consumption can cause hangovers such as thirst, fatigue / drowsiness, headache / dizziness, nausea / vomiting, helplessness, bloating / gastrointestinal discomfort. Chronic liver disease, gastritis, pancreatitis, high blood pressure, stroke, It is reported to cause many diseases such as esophagitis, diabetes and heart disease.

If you consume alcohol under normal conditions, about 10% of your body is excreted through your breath or urine, and most of the remaining 90% is metabolized in your gastric mucosa or liver. At this time, about 90-95% of the alcohol metabolism is oxidized and decomposed in the liver.

Alcohol is oxidized to alcohol dehydrogenase (ADH) and aldehyde dehydronase (ALDH) present in the liver. First, it is oxidized to acetaldehyde by ADH, which is produced by the action of aldehyde ALDH. After oxidized by Acetate, it is released into liver cells and flows through blood to tissues such as muscles, which is used for energy production. At this time, carbon dioxide (CO ₂ ) and water (H ₂ O) are produced. Increased carbon dioxide is a major factor in acidifying the blood and causes hangovers such as vomiting, headache, thirst, nausea and lethargy. Acetaldehyde, the first metabolite of alcohol accumulated in the liver, is also a high concentration of active compound, causing sweat, rapid pulse, skin flushing, nausea, vomiting, etc. For this reason, most researchers believe that acetaldehyde, the alcohol breakdown product, is the cause of hangovers.

Symptoms of a hangover begin within a few hours after drinking and last for almost a day. The experience of a particular hangover is very personal and depends on the situation and the type and amount of alcohol consumed. Symptoms of physical hangover include headache, fatigue, light and sound sensitivity, myalgia, redness of the eyes, thirst, nausea, vomiting and stomach pain. Hangover causes fast heartbeat, excitement, elevated blood pressure and sweating. Related mental symptoms of hangovers include sleep deprivation, changes in sleeping area, decreased attention, decreased concentration, depression, dizziness, anxiety and irritability.

In order to solve the hangover after drinking, bean sprouts, curry, mung bean have been used as food for hangover after drinking. In particular, bean sprouts are rich in aspartic acid, which is known to protect the liver by promoting the action of alcohol dehydrogenase (ADH) and promoting alcohol degradation. Currently, a hangover drink containing aspartic acid is also commercially available.

Recently, various drugs for reducing and eliminating hangovers after drinking have been developed, but due to their toxicity and side effects, much attention has been drawn to the development of safer natural-derived health drinks.

Accordingly, there is a trend of increasing patent applications for hangover composition using natural plant extracts.

Accordingly, the present inventors have confirmed that the chlorella-herb complex can be used as a medicament or functional food for improving liver function or hangover by confirming that the chlorella-herb complex exhibits excellent activity in improving liver function and relieving hangover.

Therefore, an object of the present invention is to provide a composition for improving liver function or hangover, comprising chlorella as an active ingredient.

In addition, another object of the present invention is to provide a health functional food for liver function improvement or hangover relief comprising chlorella as an active ingredient.

In order to achieve the above object, the present invention is a chlorella extract; And it provides a composition for improving liver function or hangover, including any one or more herbal extracts selected from the group consisting of pine needles, mulberry leaves, barley shoots, pumpkin, thorny ginseng, mugwort, eoseongcho, fresh vinegar, motherwort, dandelion and cornus as an active ingredient .

In addition, the present invention is a dry powder of chlorella; And pine needles, mulberry leaves, barley shoots, pumpkin, thorny scabies, mugwort, eoseongcho, Sinseoncho, motherwort, dandelion and cornus oil, any one or more herbs dried powder selected from the group as an active ingredient for improving liver function or hangover do.

In one embodiment of the present invention, 40 to 70 parts by weight of chlorella, 9 to 18 parts by weight of pine needles, 4.5 to 10.5 parts by weight of mulberry leaves, 3 to 6 parts by weight of barley sprout, 3 to 6 parts by weight, based on 100 parts by weight of the composition 1, 3 parts by weight, wormwood 1.5 to 1.5 parts by weight, mugwort vinegar 1.5 to 3 parts by weight, fresh vinegar 1.5 to 3 parts by weight, motherwort 1.5 to 3 parts by weight, dandelion 1.5 to 3 parts by weight, and corn coriander 1.5 to 3 parts by weight It may include.

In one embodiment of the present invention, the extract may be obtained by extraction with water or an organic solvent.

In one embodiment of the present invention, the dry powder may be powdered by drying at a temperature of less than 100 ℃.

In one embodiment of the present invention, the composition may further comprise any one or more extracts or dry powder selected from the group consisting of phosphorus mugwort, germinated brown rice, purslane, yellow mushroom and sesame powder.

In one embodiment of the present invention, it can exhibit the activity of reducing the concentration of carbon dioxide in the blood, the activity of increasing the pH of the blood and the activity of reducing the concentration of lactic acid in the blood.

In one embodiment of the invention, the composition may exhibit activity to reduce the concentration of AST, ALT and r-GTP in the blood.

In one embodiment of the present invention, the composition may reduce the activity of alcohol dehydrogenase (ADH) or reduce the activity of aldehyde dehydrogenase (ALDH).

In addition, the present invention provides a health functional food for liver function improvement or hangover relief comprising the composition as an active ingredient.

The composition for improving liver function or releasing hangover according to the present invention has an effect of reducing the blood alcohol concentration and inhibiting acidification of blood (effect of reducing the concentration of carbon dioxide and lactic acid in the blood, acidified blood pH due to alcohol intake to normal alkali). Conversion effect) is very excellent, and hangover relief effect such as protection from liver damage caused by excessive intake of alcohol, gastrointestinal function improvement effect, headache reduction effect is very excellent. In addition, since the present invention is excellent in bio stability using natural substances, it does not cause various side effects due to toxicity compared to the conventional hangover relievers, so it is useful for improving liver function or preventing hangover, eliminating hangovers and preparing functional foods. Can be used.

The present invention relates to a novel use of chlorella, and is characterized by providing a composition for improving liver function or hangover, comprising chlorella as an active ingredient.

The "chlorella" of the present invention is a freshwater algae belonging to the green algae Chlorellaaceae, and is a spherical or oval single-celled plant having a diameter of 10 µm or less, and is commonly found in rusted water and wetlands. Currently, about 10 species are known, which are more efficient than sexual reproduction such as proliferation by asexual reproduction and the increase of constituents of the body by photosynthesis, forming daughter cells, making organic matter by photosynthesis. This is characterized by high and high protein content. In particular, chlorella increases about 10 times per day under moderate conditions, so the annual production of organic matter is about 8 times that of rice. In addition, depending on the culture conditions can increase the content of fat 20 ~ 80%, protein 90%, and carbohydrates 37%, has been studied to solve the food problem of human by culturing in large quantities. However, the individual is too small, it is difficult to digest if eaten as it is, dry chlorella has a peculiar smell has a disadvantage of lowering the taste when added to food has not been developed as a food, but recently increased the absorption of digestion by sterile pure water culture Supplements and dairy products are being developed and used.

In addition, indirect use of chlorella as food or incorporation into other foods has been studied, for example, to promote the development of microorganisms such as lactic acid bacteria, or to purify contaminated water, or to mix it with livestock feed. Also used. Chlorella is not only a high-nutrition food resource of the next generation, but also has been steadily increasing all over the world as it has proved its functions such as constitution improvement, thrombosis improvement, anti-cancer effect by increased immunity, toxic neutralization and embryogenic effect of heavy metals.

However, studies on the effects of improving or inhibiting alcoholic liver damage and hangover caused by drinking chlorella are insignificant.

Therefore, the present inventors experimented with the pharmacological effect of inhibiting or improving hangover and liver dysfunction or clinical phenomenon (eg, acidification of blood) by using chlorella containing protein and chlorophyll abundantly without side effects in living body. As a result, it was found that the composition containing chlorella shows an excellent effect in improving liver dysfunction and relieving hangover through the excellent hangover inhibitory activity, liver damage inhibitory activity and blood acidification inhibitory activity.

Therefore, the present invention can provide a composition for improving liver function or relieving hangover, including chlorella as an active ingredient.

Liver function improvement or hangover composition according to the present invention is specifically chlorella extract; And one or more herbal extracts selected from the group consisting of pine needles, mulberry leaves, barley shoots, pumpkins, thorny ginsengs, mugwort, eoseongcho, fresh vinegar, motherwort, dandelion and cornus as an active ingredient, and also dry chlorella powder; And at least one herb dry powder selected from the group consisting of pine needles, mulberry leaves, barley shoots, pumpkins, thorny ginseng, mugwort, eoseongcho, fresh vinegar, motherwort, dandelion and cornus as an active ingredient.

The composition of the present invention preferably uses an extract or dry powder of chlorella and various herbal substances.

In the present invention, chlorella can be used as long as environmentally friendly natural culture, for example, crude protein is 27 to 28% of the dry weight, chlorophyll is 57.5 to 59% of the dry weight, iron is 67.5 to 69% of the dry weight, Total ferbibiide may be used that contains less than 40%, but is not limited thereto.

“Herb” as defined in the present invention refers to herbal plants that have been used as medicines or spices since ancient times, pine needles, mulberry leaves, barley sprouts, pumpkins, thorns, mugwort, eoseongcho, fresh vinegar, motherwort, dandelion, cornus, injin mugwort, germinated brown rice, Includes purslane, yellow mushrooms, and sesame powder.

"Extract" as defined in the present invention is obtained by extraction and separation from natural raw materials using extraction and separation methods known in the art and using a suitable solvent, for example, crude extracts of chlorella or natural herbs, It includes both polar solvent soluble extracts or nonpolar solvent soluble extracts.

As a suitable solvent for extracting the extract, any pharmaceutically acceptable organic solvent may be used, and water or an organic solvent may be used, but is not limited thereto. For example, purified water and methanol Alcohols containing 1 to 4 carbon atoms, acetone, ether, benzene, chloroform, including ethanol, propanol, isopropanol, isopropanol, butanol, etc. , Various solvents such as ethyl acetate, methylene chloride, hexane and cyclohexane may be used alone or in combination.

As the extraction method, any one of the methods such as hot water extraction method, cold extraction method, reflux cooling extraction method, solvent extraction method, steam distillation method, ultrasonic extraction method, elution method and compression method can be selected and used. In addition, the desired extract may be further subjected to a conventional fractionation process or may be purified using a conventional purification method. There is no limitation on the method of preparing the extract, and it is obvious that any known method may be used.

In an embodiment of the present invention, the raw chlorella and various herbs are dried and powdered at a temperature of 100 ° C. or lower, mixed at a constant ratio, and hot water extracted using purified water, but is not limited thereto.

The chlorella and various herbs have been used for a long time as a herbal medicine and edible extracts of the present invention extracted from them also have no problems such as toxicity and side effects.

As defined in the present invention, the "dry powder" may be one obtained by drying and then grinding chlorella and various herbal materials. In an embodiment of the present invention, the raw chlorella and various herbs are dried and powdered at a temperature of 100 ° C. or less, and then mixed and kneaded at a predetermined ratio to prepare granules, but is not limited thereto.

According to the present invention, extracts or dry powders of chlorella and various herbs as described above may be mixed within various composition ranges, but 40 to 70 parts by weight of chlorella, 9 to 18 parts by weight of pine needles, mulberry leaves 4.5 with respect to 100 parts by weight of the total composition. ~ 10.5 parts by weight, barley shoots 3 to 6 parts by weight, pumpkin 3 to 6 parts by weight, thorny skin 1.5 to 3 parts by weight, mugwort 1.5 to 1.5 parts by weight, fishery vinegar 1.5 to 3 parts by weight, fresh vinegar 1.5 to 3 parts by weight, motherwort 1.5 to 3 parts by weight, dandelion 1.5 to 3 parts by weight and cornus oil is preferably mixed in a composition of 1.5 to 3 parts by weight.

In addition, the composition of the present invention may further comprise any one or more extracts or dry powder selected from the group consisting of jinjin wormwood, germinated brown rice, purslane, yellow mushrooms and sesame powder.

On the other hand, the "hangover" in the present invention is a side effect that appears after alcohol intake, for example headache, dizziness, drowsiness, thirst, general boredom, fatigue, memory loss, bloating, indigestion, nausea, vomiting, diarrhea, light and sound All of the physiological side effects that are caused by blood acidity (such as sensitivity, myalgia, redness of the eyes, nausea, vitamin deficiency), acidification of the blood (lowering pH) by an increase in blood carbon dioxide (CO ₂ ), etc. Means.

In one embodiment of the present invention in the SD rat animal model by treating the chlorella-hub complex of the present invention, blood alcohol concentration, blood acidification (CO ₂ , carbonate ion, pH, lactic acid), hepatic impairment clinical indicators (AST , ALT, r-GTP), alcohol oxidase (ADH, ALDH) by examining the effects on the activity was confirmed to have an effective effect in improving liver function or hangover. As a result, the chlorella-hub composite composition of the present invention has excellent activity of reducing blood alcohol concentration (see Experimental Example 3), reduces the concentration of carbon dioxide and carbonate ions in blood, and restores blood pH from acid to normal alkali. Excellent in reducing blood lactate concentration (see Experimental Example 4), excellent in reducing blood AST, ALT and r-GTP concentrations (see Experimental Example 5), alcohol It was found that the effect of reducing the activity of dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) is excellent. Through this action it was confirmed that the chlorella-hub complex of the present invention can improve liver function and relieve hangover.

Therefore, the liver function improving or hangover composition according to the present invention is to reduce the blood alcohol concentration and the activity of inhibiting blood acidification (the effect of reducing the concentration of carbon dioxide and lactic acid in the blood, normalized blood pH due to alcohol intake) Recovery effect from alkali, very good hangover relief effect from liver damage caused by excessive intake of alcohol, gastrointestinal function improvement, headache reduction effect.

Accordingly, the liver function improving or hangover composition according to the present invention can reduce blood alcohol concentration strongly compared to the conventional hangover relievers, it can more effectively defend against liver damage that may be caused, and alcohol consumption for a long time As a result of confirming that the intake of chlorella has an effect of improving alcoholism in chronic alcoholism caused by the disease, it is proved that the possibility of development as an effective hangar relievers is sufficient.

Therefore, the composition for improving liver function or hangover, including the chlorella-herb complex according to the present invention as an active ingredient, may be used as a pharmaceutical composition.

The liver function improving or hangover composition according to the present invention may contain a pharmaceutically effective amount of chlorella-hub complex alone or may include one or more pharmaceutically acceptable carriers, excipients or diluents. The pharmaceutically effective amount herein refers to an amount sufficient to prevent, ameliorate and treat the symptoms of liver damage or hangover.

The pharmaceutically effective amount of the chlorella-hub complex according to the invention is 0.5-100 mg / day / kg body weight, preferably 1-50 mg / day / kg body weight. However, the pharmaceutically effective amount may be appropriately changed depending on the degree of liver injury or hangover symptoms, the age, weight, health condition, sex, route of administration and duration of treatment of the patient.

In addition, the pharmaceutically acceptable means a composition which is physiologically acceptable and does not cause an allergic reaction such as gastrointestinal disorders, dizziness or the like when administered to humans. Examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. Further, it may further include a filler, an anticoagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent and an antiseptic agent.

In addition, the compositions of the present invention may be formulated using methods known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to the mammal. The formulations may be in the form of powders, granules, pills, tablets, capsules, emulsions, liquids, syrups, aerosols, soft or hard gelatin capsules, sterile injectable solutions, sterile powders.

The composition for improving or relieving liver function according to the present invention may be administered through various routes including oral, transdermal, subcutaneous, intravenous or muscle, and the dosage of the active ingredient is determined by the route of administration, age, sex, weight and It may be appropriately selected depending on various factors such as the severity of the patient.

In addition, the liver function improving or hangover composition of the present invention can be administered in parallel with known compounds having the effect of preventing, improving or treating liver damage to improve liver function and relieve hangover.

Therefore, the present invention can provide a medicament capable of preventing and treating liver improvement and hangover symptoms, including a composition comprising a chlorella-herb complex as an active ingredient.

Furthermore, the liver function improvement or hangover relieving composition according to the present invention can be used as a pharmaceutical composition for improving liver function and hangover as described above, as well as the preparation of liver functional improvement and hangover rehabilitation health functional foods. It can also be used as a food composition for.

Therefore, the present invention can provide a composition for improving liver function and relieving hangover, including a chlorella-herb complex as an active ingredient.

The food composition according to the present invention can be easily utilized as a food, for example, a main ingredient, an auxiliary ingredient, a food additive, a functional food or a drink, which is effective in improving liver function and relieving hangover.

As used herein, the term “food” refers to a natural product or processed product containing one or more nutrients, and preferably means a state in which it can be directly eaten through a certain processing step, It includes all foods, food additives, functional foods and drinks.

Foods to which the composition for food for improving liver function and hangover according to the present invention can be added include, for example, various foods, beverages, gums, teas, vitamin complexes, functional foods, and the like. In addition, in the present invention, the food may include special nutritive foods (e.g., crude oil, spirits, baby food, etc.), meat products, fish meat products, tofu, mackerel, noodles (Such as soy sauce, soybean paste, kochujang, mixed potatoes), sauces, confectionery (eg, snacks), candies, chocolate, gums, ice cream, milk products (eg, fermented milk, cheese, But are not limited to, pickled foods (various kinds of kimchi, pickles, etc.), beverages (e.g., fruit drinks, vegetable beverages, beverages, fermented beverages and the like) and natural seasonings (e.g. The food, beverage or food additive may be prepared by a conventional production method.

The above-mentioned " functional food " refers to a food group which is imparted with added value to function and express the function of the food by physical, biochemical or biotechnological techniques, or to control the bio-defense rhythm of the food composition, Refers to a food prepared by processing a body so as to sufficiently express the body's control function on the body, such as recovery, and the like. Specifically, it may be a health functional food. The functional food may include a food-acceptable food-aid additive, and may further comprise suitable carriers, excipients and diluents conventionally used in the production of functional foods.

In addition, in the present invention, the "beverage" refers to a generic term for drinking to quench thirst or to enjoy a taste and includes a functional drink. The beverage contains, as an essential ingredient, the composition for improving the liver function or hangover, as an essential ingredient, and there are no special limitations on the other ingredients, and various flavors or natural carbohydrates as additional ingredients are included as in the general beverage. can do.

Furthermore, in addition to the above, foods containing the composition for improving liver function or hangover of the present invention include various nutrients, vitamins, minerals (electrolytes), synthetic flavors, and natural flavoring agents, such as colorants and fillers (cheese). , Chocolate and the like), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. The components can be used independently or in combination.

Therefore, the present invention can provide a dietary supplement comprising a chlorella-herb complex and a food supplement acceptable food supplement additive exhibiting an effect of improving liver function or hangover.

The type of dietary supplement is not limited thereto, but may be in the form of powder, granules, tablets, capsules, or beverages.

In addition, in a food containing a composition for food for improving liver function or relieving hangover of the present invention, the amount of the composition according to the present invention may comprise from 0.001% to 90% by weight of the total food weight, preferably May be included in an amount of 0.1% by weight to 40% by weight, and in the case of a beverage, it may be included in a ratio of 0.001g to 2g, preferably 0.01g to 0.1g based on 100ml, but for health and hygiene purposes In the case of prolonged ingestion for the purpose of health control, the range may be below the above range, and the active ingredient is not limited to the above range because the active ingredient may be used in an amount above the above range because there is no problem in terms of safety.

Hereinafter, the present invention will be described in detail with reference to embodiments and drawings. However, these examples are intended to illustrate the present invention in more detail, and the scope of the present invention is not limited to these examples.

< Manufacturing example  1>

<1-1> Preparation of materials

Chlorella Co., Ltd. purchased Chlorella produced by eco-friendly culture / manufactured by Korea Chlorella Co., Ltd. (81-3, Challenge-ri, Maam-myeon, Goseong-gun, Gyeongnam).

Herbs used pine needles, mulberry leaves, barley sprouts, pumpkins, thorns, wormwood, eoseongcho, fresh vinegar, motherwort, dandelion, cornus as raw materials recognized as food in the Korean Food Code.

<1-2> Preparation of Chlorella-Herb Mixture

In order to prepare a chlorella-hub mixture, first, chlorella and various herbs were dried at 80 ° C. for 6 hours using a hot air dryer (Jeju Pharm. Korea, Korea). Each dried raw material was pulverized to about 180 mesh using a pin mill grinder (Jeong Pharm. Food Machinery, Korea). Four times purified water was added to each of the pulverized raw materials, and extracted with hot water at 80 ° C. for 7 hours using an extracting concentrator (Dongnam Industrial, Korea). Korea) and each extract was mixed in the ratio shown in Table 1 below to prepare a chlorella-hub mixture.

% Of each extract Raw material name Formulation ratio (%) Chlorella 40 to 70 pine needles 9 to 18 Mulberry leaf 4.5 to 10.5 Barley grass 3 to 6 pumpkin 3 to 6 A goose 1.5 to 3 Mugwort 1.5 to 1.5 Sophora 1.5 to 3 Chinese 1.5 to 3 motherwort 1.5 to 3 dandelion 1.5 to 3 Corn oil 1.5 to 3

< Manufacturing example  2>

Chlorella-Herb Dry Powder (Granule) Preparation

Raw chlorella and various herbs presented in Preparation Example 1, namely, chlorella, pine needles, mulberry leaves, barley sprouts, pumpkins, thorny ginseng, mugwort, eoseongcho, fresh vinegar, motherwort, dandelion and cornus oil after drying at less than 100 ℃, dried chlorella And the herbs were ground in a mill. The pulverized chlorella and herbs were mixed and kneaded in the proportions listed in Table 1 above, and the mixed dough was formulated with a granulator and dried.

< Experimental Example  1>

Evaluation of Stability of Chlorella-Hub Mixtures

The inventors of the present invention, in order to evaluate the stability of the chlorella-herb mixture prepared in Preparation Example 1 and the chlorella-herb dry powder prepared in Preparation Example 2, the number of chlorella cells, E. coli, general bacteria, chlorophyll, crude protein and vitamins, etc. Was analyzed.

<1-1> Chlorella Number of Cells  Measure

The chlorella-herb mixture was observed under a microscope (Olympus Optical Co., Ltd., Japan) on a Haemacytometer (Erma Co., Ltd., Japan).

<1-2> Escherichia coli  Measure

1 ml of sample was inoculated into 3M Petrifilm ^TM E. coli / Coliform Count Plate (3M Microbiology Products, USA) and placed in an incubator (CO ₂ Incubator J 2000, Jisico, Co., Ltd., Korea) and incubated at 37 ° C. for 24 hours. After that, red colonies were identified by E. coli / Coliform and colony counts were calculated.

<1-3> General bacteria count

3M Petrifilm ^™ Aerobic Count Plate (3M Microbiology Products, USA), 3M Petrifilm ^™ Rapid Stap. aureus Count Plate (3M Microbiology Products, USA) and 3M Petrifilm ^TM Yeast and Mold Count Plate incubator (CO ₂ Incubator J 2000, Jisico according to the manual using a 3M (3M Microbiology Products, USA), respectively, Co., Ltd., Cultivated in Korea) and the colonies were calculated and added together.

<1-4> Chlorophyll

Samples were extracted with acetone, separated at 3500 rpm with a centrifuge (MF80, Hanil Science, Korea), and then absorbed at A645 and A663 using UV-Visible Spectrophotometers (Nicolet Evolution 300, Thermo Electron Spectroscopy Ltd., UK). It was.

<1-5> Crude protein  And measurement of vitamins

Crude protein and vitamin content were measured by requesting from Dongjin Life Research Institute (Changwon, Gyeongnam).

Analysis of stability evaluation of chlorella-herb mixture Item Chlorella-Herb Mixture Chlorella-Herb Dry Powder Cell number - - Coliform group (negative) voice voice Total microbial water (/ ml) Below 100 - Crude protein (%) 0.31 25.1 Tar color - Not detected Chlorophyll (mg / 100g (ml)) 545.8 1231.9 Vitamin B2 (mg / 100g) - 1.2 Preservative (g / kg) 0.002 - Total Pheophorbide (mg / 100g) ^* - 0.0 Lead (mg / kg) ^* Not detected - Cadmium (mg / kg) ^* Not detected - Iron (mg / 100g) ^* - 90.4

* Analysis Source: Ltd. Korea Dongjin Chlorella Research Center and Institute of Life ^*

< Experimental Example  2>

Animal Administration and Blood Collection

The present inventors administered to the animals as follows to determine how the chlorella-hub mixed solution prepared in Preparation Example 1 affects the hangover phenomenon caused by alcohol, and collected blood for use in the test.

<2-1> Administration method

375 to 395 g SD rats were orally administered with chlorella-hub mixture and conventional hangover relievers (C products, Y products, M products) to 10 ml / kg-BW and after 30 minutes, 40% alcohol was 10 ml / kg-BW Orally administered to As a normal control, water was orally administered to 10 ml / kg-BW, and 30 minutes later, 40% alcohol was orally administered to 10 ml / kg-BW.

<2-2> Blood Collection

① Blood sampling for blood alcohol concentration measurement : 600 ~ 1000ml of blood was collected from tail vein before and after 1 hour, 3 hours, 5 hours after alcohol administration, and the collected blood was centrifuged at 3000 rpm for 10 minutes for upper layer. Serum was obtained. After measuring the alcohol concentration of the obtained serum, the trend was confirmed.

② Blood collection for blood CO ₂ and pH measurement : 24 hours after alcohol administration, open the blood and collect the blood from the abdominal aorta using a heparin-treated syringe, and then collect the blood immediately at 3000 rpm for 10 minutes. The upper plasma was obtained by centrifugation and used for measuring CO ₂ and pH.

③ Blood sampling for measuring lactate in blood : 3M after taking alcohol and taking blood from the abdominal vein after 24 hours, 3M immediately to prevent the increase of lactic acid as a result of glucose metabolism. Perchloric acid (perchloric acid) was treated and centrifuged at 3000rpm for 10 minutes to obtain the upper protein-retained serum was used for lactic acid measurement.

④ Blood AST , ALT, and r- GTP measurement blood collection : 24 hours after alcohol administration, open the blood and collect the blood from the abdominal vein and centrifuged at 3000rpm for 10 minutes Were obtained and used for AST, ALT and r-GTP measurements.

< Experimental Example  3>

Chlorella-Herb Mixture in Blood Alcohol  Impact on concentration

The present inventors compared and analyzed the effect of the chlorella-hub mixed solution prepared in Preparation Example 1 of the present invention and the existing hanger on the change of blood alcohol concentration, which is one of the hangover phenomena. To this end, the alcohol concentration kit (BioAssay-ECET100) was measured from the serum separated in Experimental Example <2-2> and the change in serum alcohol concentration was measured.

The alcohol measurement kit follows a principle of measuring and quantifying formazan MTT chromogen by 565 nm absorbance by NADH, which is formed by alcohol dehydrogenase oxidizing alcohol. 10 μl of the prepared serum was loaded into a 96-well plate, 90 μl of the reaction reagent was added thereto, and placed at room temperature for 30 minutes. Then, 100 μl of the stop reagent was added to stop the reaction.

As a result, as shown in Table 3, the water + alcohol intake group as a control group showed that the blood alcohol concentration increased to 10.12 ± 1.23 (mM) after 3 hours after alcohol intake and then decreased slightly after 5 hours. In addition, the chlorella-herb mixture + alcohol intake group was confirmed that the blood alcohol concentration is maintained low at any time after 1 hour, 3 hours, 5 hours of alcohol intake.

These results indicate that the chlorella-hub mixture according to the present invention has an effect of reducing the alcohol concentration in the blood increased by ingestion of alcohol. In particular, the chlorella-herb mixture was found to have an effect of more strongly reducing the alcohol concentration in the blood than conventional hangover C products, Y products and M products. It was speculated that the chlorella-hub mixture would lower the alcohol concentration in the blood because it inhibited or delayed absorption of the alcohol into the blood.

Results of Stability Assessment Analysis of Chlorella-Herb Mixture Water + alcohol Chlorella-Herbs Mixture + Alcohol C product + alcohol Y product + alcohol M product + alcohol Alcohol (mM) Alcohol (mM) Alcohol (mM) Alcohol (mM) Alcohol (mM) Before alcohol 0 ± 0.01 0 ± 0.07 0 ± 0.07 0 ± 0.07 0 ± 0.07 1 hour after alcohol 9.23 ± 2.46 2.57 ± 0.04 ^* 6.40 ± 0.29 6.55 ± 0.23 6.15 ± 0.13 3 hours after alcohol 10.12 ± 1.23 6.12 ± 0.28 ^** 6.70 ± 0.15 ^* 8.39 ± 0.10 ^* 8.43 ± 0.07 5 hours after alcohol 8.29 ± 1.61 4.59 ± 0.06 ^* 5.11 ± 0.16 ^* 6.52 ± 0.13 5.24 ± 0.13 ^*

Data are expressed as mean ± SD (n = 3), where * indicates p <0.05 and ** indicates p <0.001 compared to the water + alcohol group.

< Experimental Example  4>

Effect of Chlorella-Herb Mixture on Acidification of Blood

The present inventors, in order to confirm the effect of the chlorella-hub mixture prepared in Preparation Example 1 of the present invention on the acidification of blood, the concentration of carbon dioxide (CO ₂ ) in the blood, the concentration of carbonate ion (HCO ₃ ⁻ ), blood pH, The changes in the concentration of lactic acid in the blood were examined.

<4-1> carbon dioxide in the blood ( CO ₂ Effect on concentration

Carbon dioxide (CO ₂ ) in the blood is the final product produced after cells produce the energy necessary for their own survival by using organic materials such as glucose. When CO ₂ increases in the blood, the pH of the blood becomes acidified. Produces respiratory disorders, and migraines. In addition, CO ₂ in the blood combines with water (H ₂ O) to form H ₂ CO ₃ (carbonate). CO ₂ + H ₂ O → H ₂ CO _{3 It} becomes. Examination of this chemical reaction shows that as CO ₂ increases, the consumption of H ₂ O increases.

Consumption of water causes thirst, and thirst after ingestion of alcohol results in increased consumption of H ₂ O by an increase in CO ₂ . Thus, changes in the concentration of CO ₂ in the blood are clinical indicators of changes in pH, blood consumption, thirst, respiratory distress and migraine headaches.

Therefore, the present inventors compared and analyzed the effect of the chlorella-hub mixture and the conventional hangover relieving agent on the increase in blood carbon dioxide concentration by the ingestion of alcohol. To this end, using a CO ₂ assay kit (Randox-CD129) from the blood plasma collected 24 hours after the administration of alcohol in Experimental Example <2-2> was measured in the blood concentration of CO _2.

The CO ₂ assay kit follows the principle of quantifying by measuring 340 nm absorbance the amount of decrease in NADH that is reduced when oxaloacetate is reduced in proportion to bicarbonate in the blood and reduced to malate. 1 ml of the reaction reagent was added to 5 µl of the prepared plasma and reacted at 37 ° C. for 5 minutes, and the absorbance was measured at 340 nm.

As a result, as shown in Table 4, the alcohol intake group (water + alcohol intake group) is the CO _{2 in} the blood of the normal control group that did not consume alcohol It was found that 20.43 ± 0.77mM existed by 8% increase from the concentration (18.92 ± 0.16mM), but the hunger killer (C product, Y product, M product) group and chlorella-hub mixed group group CO _{2 in the} blood The concentration is CO ₂ in the blood of the alcoholic group. It was found to be maintained at a concentration lower than the concentration (20.43 ± 0.77 mM). In particular, when the chlorella-hub mixture of the present invention is ingested, CO _{2 in the} blood is lowered to a concentration lower than the control group's CO ₂ concentration (18.92 ± 016mM) (18.03 ± 0.72mM). It was found that the concentration was lowered, and this resulted in the strongest effect of reducing the concentration of CO ₂ in the blood after alcohol consumption.

Change The results of the CO ₂ concentration in blood CO ₂ concentration (mM) CO ₂ increase(%) Normal control 18.92 ± 0.16 0 - Water + alcohol 20.43 ± 0.77 +8.0 0 Chlorella-Herb Mixture + Alcohol 18.03 ± 0.72 ^* - -11.74 C product + alcohol 19.95 ± 1.56 - -2.36 Y product + alcohol 18.97 ± 0.81 ^* - -7.14 M product + alcohol 18.62 ± 0.70 ^* - -8.86

Data are expressed as mean ± SD (n = 3), where * indicates p <0.05 and ** indicates p <0.001 compared to the water + alcohol group.

<4-2> blood Carbonate ions (HCO ₃ ^- )of  Impact on concentration

On the other hand, carbonate ions in the blood (HCO ₃ ^-) is proportional to the concentration of CO ₂ in the blood, and this _{CO 2 + H 2 O → H} 2 CO 3 → HCO 3 - can be seen from the chemical equilibrium reactions + H ^+. Carbonate ion (HCO ₃ ^-) concentration was calculated from the following equation 1.

(Equation 1)

As a result, as shown in Table 5 below, the alcohol intake group was found to be 7.8% higher than the carbonate concentration (18.61 ± 0.15 mM) of the normal control group, indicating that 20.06 ± 0.76 mM carbonate was present. It reflects the result that the concentration of CO ₂ increased serum than blood levels of CO ₂ in the control group in the group. However, chlorella-hub mixtures have been shown to strongly reduce the concentration of carbonate ions increased by alcohol intake. This result is consistent with the effect of the chlorella-hub mixture having the strongest reduction in the concentration of carbon dioxide in the blood (see Table 4).

Analysis result of change in blood carbonate concentration HCO ₃ ^- Concentration (mM) increase(%) HCO ₃ ^- increase or decrease in concentration (pM) Normal control 18.61 ± 0.15 0 - 0 - Water + alcohol 20.05 ± 0.76 +7.8 0 +1440 0 Chlorella-Herb Mixture + Alcohol 17.74 ± 0.71 ^** - -11.6 - -2310 C product + alcohol 19.61 ± 1.53 - -2.2 - -440 Y product + alcohol 18.65 ± 0.79 ^* - -7.0 - -1400 M product + alcohol 18.28 ± 0.68 ^* - -8.9 - -1770

Data are expressed as mean ± SD (n = 3), where * indicates p <0.05 and ** indicates p <0.001 compared to the water + alcohol group.

<4-3> blood pH Effect on

The pH of the blood is changed by the concentration of carbon dioxide (CO ₂ ) and carbonate (HCO ^{3 −} ) in the blood. As can be seen from the pH calculation formula of Equation 1, when the CO ₂ concentration in the blood is increased by the ingestion of alcohol, and the HCO ^{3 −} concentration is increased, the pH of the blood is changed to acidic. The clinical symptoms of such a change in blood pH are called respiratory acidosis, in which case respiratory distress and headaches occur.

Therefore, the present inventors compared and analyzed the effect of the chlorella-hub mixture and the existing hangover relievers on the blood pH change by the intake of alcohol. To this end, blood pH was measured using a pH meter (Schott-SRDC22N: 1010) from plasma collected 24 hours after alcohol administration in Experimental Example <2-2>.

As a result, as shown in Table 6, the blood pH of the normal control group is 7.88 ± 0.01, but in the alcohol intake group, the pH was decreased to 7.83 ± 0.01 by the ingestion of alcohol, and the pH of the blood was changed to acidic. Due to this, the blood hydrogen ion (H ⁺ ) concentration is increased by 1610pM than the blood hydrogen ion (H ⁺ ) concentration (13.18 nM) of the normal control, reflecting the results shown as 14.79 nM. However, ingestion of chlorella-herb mixture and conventional hangover relievers (C and Y products) increased the pH of the acidified blood by alcohol by decreasing the concentration of hydrogen ions increased by alcohol intake. Appeared to recover. In particular, the chlorella-hub mixture has been shown to restore acidified pH (7.83 ± 0.01) to 7.88 ± 0.02, similar to the pH of the normal control group (7.88 ± 0.01) by alcohol intake. , Y product, M product) was found to reduce the concentration of hydrogen ions more effectively.

Analysis of changes in blood pH pH H ⁺ concentration (nM) Ratio to control Change in H ⁺ concentration (pM) Normal control 7.88 ± 0.01 13.18 One 0 - Water + alcohol 7.83 ± 0.01 14.79 1.122 +1610 0 Chlorella-Herb Mixture + Alcohol 7.88 ± 0.02 ^* 13.18 One - -1610 C product + alcohol 7.87 ± 0.02 ^* 13.49 1.023 - -1300 Y product + alcohol 7.86 ± 0.02 ^* 13.80 1.047 - -990 M product + alcohol 7.83 ± 0.01 14.79 1.122 - 0

Data are expressed as mean ± SD (n = 3), where * indicates p <0.05 and ** indicates p <0.001 compared to the water + alcohol group.

<4-4> Lactic acid in the blood ( lactate Effect on concentration

When intake of alcohol and consume a large amount (glucose) sugar in the brain, the resulting concentration than in normal sugar in the blood at the same time as increasing the blood in CO ₂ concentration by being the generation of CO ₂ increases the pH of the blood acidified by Lowering hypoglycemia occurs. In order to compensate for the hypoglycemic phenomenon, a reaction to generate new sugars in vivo proceeds, and lactate is used to generate new sugars. Increasing the amount of lactic acid in the blood increases the concentration of hydrogen ions (H ⁺ ) in the blood, which causes the pH of the blood to become acidic. This pathology is called lactic acidosis, which causes vomiting, dizziness, respiratory distress, You will have a headache. This clinical symptom is also one of the hangover phenomenon, the present inventors measured the lactic acid concentration in the blood.

Therefore, the present inventors compared and analyzed the effect of the chlorella-hub mixture and the existing hangover relievers on the blood pH change by the intake of alcohol. To this end, the concentration of lactic acid was measured from the deproteinized serum collected in Experimental Example <2-2>.

The amount of NADH formed as lactic acid was oxidized by lactate dehydrogenase was measured by 340 nm absorbance. Absorbance was measured at.

As a result, as shown in Table 7 below, 40.40 ± 3.37 (μM) of lactic acid was present in the blood of the normal control group, but the alcohol intake group increased 47.7% than the control group by alcohol intake, resulting in 59.66 ± 6.07 (μM) lactic acid. Appeared to be. However, chlorella-hub mixtures and conventional hangover relievers (C, Y and M products) have been shown to reduce blood lactate concentrations increased by alcohol consumption. In particular, the chlorella-hub mixed solution of the present invention showed a lower lactic acid concentration than the normal control, it was confirmed that the effect of effectively maintaining the blood lactate concentration low.

Analysis result of change in blood lactate concentration lactate concentration (μM) increase or decrease lactate (%) lactate concentration
Increase / Decrease (μM) Normal control 40.40 ± 3.37 0 - 0 - Water + alcohol 59.66 ± 6.07 +47.7 0 19.26 0 Chlorella-Herb Mixture + Alcohol 36.77 ± 1.97 ^** - -38.9 - -22.89 C product + alcohol 44.45 ± 1.83 ^* - -25.5 - -15.21 Y product + alcohol 24.94 ± 0.44 ^** - -58.2 - -34.72 M product + alcohol 44.28 ± 4.28 ^* - -25.8 - -15.38

Data are expressed as mean ± SD (n = 3), where * indicates p <0.05 and ** indicates p <0.001 compared to the water + alcohol group.

< Experimental Example  5>

Effect of Chlorella-Herb Mixture on Clinical Indicators of Liver Disorder

The present inventors examined the changes in blood AST, ALT, r-GTP in order to determine the effect of the chlorella-hub mixed solution prepared in Preparation Example 1 of the present invention on clinical indicators of liver failure.

<5-1> blood AST Effect on the activity of

AST transfers the amino group (-NH ₂ ) in aspartic acid to the carbonyl group (C = O) of alpha-ketoglutaric acid, and aspartic acid is oxaloacetic acid. Glutaric acid turns into glutamic acid. When the cell membrane is damaged by the oxidation of hepatic phospholipids, AST in the liver cells is secreted into the blood. Therefore, blood AST is used as an indicator of liver damage. Clinical symptoms and degeneration of NADH / NADPH / glutathione result in coma due to ammonia accumulation due to cell and body deterioration and urea degeneration without toxicity.

Therefore, the present inventors compared and analyzed the effects of the chlorella-hub mixture and the conventional hangover on the change of AST, one of the indicators of liver damage. To this end, blood AST was measured from the serum collected in Experimental Example <2-2> using AceChem AST kit (Dongdong-A301). The measurement kit follows the principle of kinetic measurement at 340 nm absorbance to reduce the amount of NADH that decreases when reducing oxalacetate produced by blood AST to malate.

1 mL of the reaction reagent was added to 100 μl of the prepared serum and reacted at 37 ° C. for 2 minutes, followed by kinetic measurement of absorbance at 340 nm for 2 minutes.

As a result, as shown in Table 8 below, the alcohol intake group was 109.32 ± 4.36 (U / L), which was 173% higher than the 40.07 ± 0.51 (U / L) of the normal control group, and the chlorella-herb mixture was 65.42 ± 1.77 (U). / L) reduced the blood AST concentration increased by 40.16% by alcohol intake. In particular, in the effect of reducing blood AST concentration, chlorella-herb mixture was found to reduce blood AST by more than 14% more strongly than conventional products C, Y and M products. Through this, it was confirmed that the chlorella-herbal mixture of the present invention is superior in the liver protection effect compared to the conventional hangover relievers (C products, Y products, M products).

Analysis of Changes in Blood AST Activity AST active (U / L) AST increase / decrease (%) Normal control 40.07 ± 0.51 0 - Water + alcohol 109.32 ± 4.36 +173 0 Chlorella-Herb Mixture + Alcohol 65.42 ± 1.77 ^** - -40.16 C product + alcohol 80.74 ± 0.51 ^** - -26.14 Y product + alcohol 84.86 ± 0.88 ^** - -22.37 M product + alcohol 120.52 ± 2.04 ^* - +10.2

Data are expressed as mean ± SD (n = 3), where * indicates p <0.05 and ** indicates p <0.001 compared to the water + alcohol group.

<5-2> blood ALT Effect on the activity of

ALT transfers the amino group (-NH ₂ ) in alanine to the carbonyl group (C = O) of alpha ketoglutaric acid, converting alanine to pyruvic acid and alpha ketoglutaric acid to glutamic acid. When the cell membrane is damaged by the oxidation of hepatocellular phospholipid, ALT in the liver cells is secreted into the blood, so blood ALT is used as an indicator of liver damage. The coma is caused by the accumulation of ammonia due to the deterioration of the cell and body due to the clinical symptoms and the reduced production of NADH / NADPH / glutathione and the production of urea which is not toxic.

Therefore, the present inventors compared and analyzed the effect of the chlorella-hub mixture and the conventional hangover on the change of ALT which is one of the indicators of liver damage. To this end, blood ALT was measured from the serum collected in Experimental Example <2-2> using AceChem ALT kit (Dongdong-A302). The measurement kit follows the principle of kinetic measurement of the amount of reduction of NADH, which is reduced when reducing pyruvate produced by blood ALT to lactic acid, at 340 nm absorbance.

1 mL of the reaction reagent was added to 100 μl of the prepared serum and reacted at 37 ° C. for 2 minutes, followed by kinetic measurement of absorbance at 340 nm for 2 minutes.

As a result, as shown in Table 9, in the alcohol intake group, 59.23 ± 3.85 (U / L) increased by 112% than the 27.99 ± 1.02 (U / L) of the normal control, Chlorella-Herb mixture 32.12 ± 0.51 (U / L) reduced the blood ALT concentration increased by 45.77% by alcohol intake. In particular, in the effect of reducing blood ALT concentration, chlorella-herb mixture was found to reduce blood ALT by more than 15% more powerfully than conventional products C, Y and M. This is consistent with the effect of chlorella-hub mixture on reducing blood AST concentrations (see Table 8). Through this, it was confirmed that the chlorella-hub mixed solution of the present invention is superior in the liver protection effect compared to the conventional hangover relievers (C products, Y products and M products).

Analysis of changes in blood GPT (ALT) activity GPT active (U / L) increase (%) Normal control 27.99 ± 1.02 0 - Water + alcohol 59.23 ± 3.85 + 111.6 0 Chlorella-Herb Mixture + Alcohol 32.12 ± 0.51 ^** - -45.77 C product + alcohol 41.84 ± 1.02 ^** - -29.36 Y product + alcohol 41.25 ± 1.84 ^** - -30.35 M product + alcohol 64.83 ± 2.70 - + 9.4

Data are expressed as mean ± SD (n = 3), where * indicates p <0.05 and ** indicates p <0.001 compared to the water + alcohol group.

<5-3> blood r- GTP Effect on the activity of

r-GTP contains ASTs such as alanine, aspartic acid and glutamic acid in the blood, substrate amino acids of ALT, constituent amino acids of antioxidant glutathione, cysteine, glycine, glutamic acid, and amino acids necessary for the synthesis of all proteins into the interior of liver cells. It is important to transport. When the cell membrane is damaged by the oxidation of hepatocellular phospholipids, r-GTP in the liver cells is secreted into the blood, so blood r-GTP is used as an indicator of liver damage. And coma due to ammonia accumulation due to clinical symptoms such as fatigue, decreased cellular and body function due to NADH / NADPH / glutathione production, and reduced urea production without toxicity.

Therefore, the present inventors compared and analyzed the effects of chlorella-hub mixture and conventional hangover on the change of r-GTP which is one of the indicators of liver damage. To this end, blood r-GTP was measured from the serum collected in Experimental Example <2-2> using AceChem r-GTP kit (Dong-A319). The measurement kit follows the principle of calculating the amount of p-nitroaniline produced by r-GTP in the blood by kinetic measurement at 405 nm absorbance and calculating it by enzyme reaction.

1 mL of the reaction reagent was added to 500 μl of the prepared serum and reacted at 37 ° C. for 2 minutes, followed by kinetic measurement at 405 nm for 2 minutes.

As a result, as shown in Table 10, in the alcohol intake group, 13.43 ± 0.61 (U / L) was increased by 22.5% than 10.96 ± 0.61 (U / L) of the normal control, the chlorella-hub mixture is 10.87 ± 0.53 (U / L) reduced the blood r-GTP concentration increased by 19.06% by alcohol intake. In particular, in the effect of reducing the concentration of r-GTP in the blood, chlorella-herb mixture was found to reduce blood r-GTP more than 8% more strongly than the existing products of hangover relievers C, Y and M products. It is similar to the r-GTP concentration of the normal control. Through this, it was confirmed that the chlorella-hub mixed solution of the present invention is superior in the liver protection effect compared to the conventional hangover relievers.

Analysis of Changes in Blood r-GTP Activity r-GTP activity (U / L) increase (%) Normal control 10.96 ± 0.61 - - Water + alcohol 13.43 ± 0.61 + 22.5 0 Chlorella-Herb Mixture + Alcohol 10.87 ± 0.53 ^** - -19.06 C product + alcohol 12.73 ± 1.37 - -5.24 Y product + alcohol 13.26 ± 0.75 - -1.29 M product + alcohol 12.37 ± 1.62 - -7.87

Data are expressed as mean ± SD (n = 3), where * indicates p <0.05 and ** indicates p <0.001 compared to the water + alcohol group.

Therefore, these results indicate that chlorella products reduce blood alcohol levels more strongly than conventional hangover cure agents, and thus more effectively defend against liver damage that may be caused by chlorella products. Zero proved the possibility of development.

< Example  6>

Effect on Liver Alcohol Oxidase Activity

When alcohol enters the liver through the blood vessels through the blood vessels, it is oxidized to acetaldehyde by alcohol dehydrogenase (hereinafter referred to as 'ADH'), and NAD ⁺ , a coenzyme of ADH, is reduced to NADH. . This acetaldehyde is oxidized to acetate by aldehyde dehydrogenase (hereinafter referred to as 'ALDH'), whereby NADH is produced from NAD ⁺ .

 Increased activity of ADH and ALDH is a feedback phenomenon to prevent the accumulation of acetaldehyde. Long-term alcohol consumption increases the activity of ADH and ALDH to decompose alcohol in the liver. Therefore, the activity of ADH and ALDH is used as an important indicator of chronic alcoholism.

Therefore, the present inventors used the chlorella-hub granules prepared in Preparation Example 2 to confirm the dietary effect of chlorella granules on the changes of alcohol oxidase (dehydrogenase) in the liver due to long-term (4 weeks) alcohol consumption. The following dietary experiments were conducted.

<6-1> Diet of Chlorella Granules and Administration of Alcohol

8-week-old SD rats were fed a diet diet and water (control), a feed containing 3% chlorella granules, and a diet and drinking water (3% chlorella intake), a diet and 25% diet. There were four groups of alcohol-drinks (25% alcohol) and 3% chlorella granules and 25% alcohol (3% Chlorella + 25% alcohol). The diet was conducted for 4 weeks.

<6-2> alcohol dehydrogenase ( ADH ) Effect on activity

We measured changes in ADH activity in the liver to ascertain the effect of the chlorella diet on the inhibition of chronic alcoholism that occurs upon prolonged alcohol intake. The measurement of ADH follows the principle of kinetic measurement at 440 nm absorbance that p-nitrosodiumethylaniline (NDMA) is reduced by oxidizing n-butanol by ADH and reduced color development.

1.9 ml of the reaction reagent was added to 100 µl of the prepared sample, and the absorbance at 440 nm was measured by kinetic for 5 minutes.

As a result, as shown in Table 11 below, in the 25% alcohol intake group, 6.97 ± 0.27 (nmol / minmg protein) increased by 40.2% from 4.97 ± 0.97 (nmol / minmg protein) in the control group, and 3% chlorella + 25% alcohol. The intake group was 4.03 ± 0.09 (nmol / minmg protein), indicating that ADH activity in the liver was completely reduced to normal due to chlorella's diet.

Analysis of changes in blood ADH activity ADH activity (nmol / minmg protein) Control group 4.97 ± 0.97 3% Chlorella 3.64 ± 0.70 25% alcohol 6.97 ± 0.27 3% Chlorella + 25% Alcohol 4.03 ± 0.09 ^**

Data presented as mean ± SD (n = 3), ** indicates p <0.001 compared to 25% alcohol intake group.

<6-3> between Aldehyde  Effect on oxidase (dehydrogenase) activity

We measured changes in ALDH activity in the liver to ascertain the effect of the chlorella diet on the inhibition of chronic alcoholism that occurs upon prolonged alcohol intake. ALDH was measured by kinetic measurement of fluorescence resulting from the reduction of 6-methoxy-2-naphthaldehyde to 6-methoxy-2-naphthoic acid, a fluorine when acetaldehyde was oxidized by ALDH. Follow the principle of calculation.

2.94 ml of reaction reagent was added to 60 µl of the prepared sample, and fluorescence was measured by kinetic for 3 minutes at excitation 310 nm / emission 360 nm.

As a result, as shown in Table 12 below, ALDH activity in the 25% alcohol intake group was 0.288 ± 0.016 (nmol / minmg protein), 28.6% higher than 0.224 ± 0.011 (nmol / minmg protein) in the control group, and 3% ALDH activity in the Chlorella + 25% alcohol intake group was 0.238 ± 0.010 (nmol / minmg protein). This means that the activity of ALDH, which was 28.6% higher than that of the control group by 25% alcohol intake, was completely inhibited due to the diet of chlorella.

Analysis result of change of blood ALDH activity ALDH activity (nmol / minmg protein) Control group 0.224 ± 0.011 3% Chlorella 0.194 ± 0.039 25% alcohol 0.288 ± 0.016 3% Chlorella + 25% Alcohol 0.238 ± 0.010 ^**

Data presented as mean ± SD (n = 3), ** indicates p <0.001 compared to 25% alcohol intake group.

Therefore, it was confirmed from the above results that the intake of chlorella in chronic alcoholism caused by long-term alcohol intake has an effect of improving alcoholism.

So far I looked at the center of the preferred embodiment for the present invention. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (11)

Chlorella extract; And
Pharmaceutical composition for improving liver function or hangover, comprising a herb extract consisting of pine needles, mulberry leaves, barley shoots, pumpkin, thorny ginseng, mugwort, eoseongcho, fresh vinegar, motherwort, dandelion and cornus as an active ingredient. Dry powder of chlorella; And
Pharmaceutical composition for improving liver function or hangover, comprising a herb dry powder consisting of pine needles, mulberry leaves, barley shoots, pumpkin, thorny ginseng, mugwort, eoseongcho, fresh vinegar, motherwort, dandelion and cornus as an active ingredient. 3. The method according to claim 1 or 2,
Chlorella 40 to 70 parts by weight, pine needles 9 to 18 parts by weight, mulberry leaves 4.5 to 10.5 parts by weight, barley shoots 3 to 6 parts by weight, pumpkin 3 to 6 parts by weight, thorny skin 1.5 to 3 parts, Mugwort 1.5 ~ 1.5 parts by weight, Eoseongcho 1.5 ~ 3 parts by weight, fresh vinegar 1.5 ~ 3 parts by weight, motherwort 1.5 ~ 3 parts by weight, dandelion 1.5 ~ 3 parts by weight and cornus milk 1.5 ~ 3 parts by weight liver improvement or hangover Relieving pharmaceutical composition. The method of claim 1,
The extract is characterized in that obtained by extracting with water or an organic solvent, liver function improvement or hangover pharmaceutical composition. 3. The method of claim 2,
The dried powder is dried and dried at a temperature of less than 100 ℃ liver function improvement or hangover pharmaceutical composition characterized in that powdered. delete 3. The method according to claim 1 or 2,
The composition of the present invention has an activity for reducing the concentration of carbon dioxide in the blood, an activity for increasing the pH in the blood, and a blood composition for improving liver function or hangover, characterized in that it has an activity for decreasing the concentration of lactic acid in the blood. 3. The method according to claim 1 or 2,
The composition is characterized in that it has an activity to reduce the concentration of AST, ALT and r-GTP in the blood liver improvement or hangover pharmaceutical composition. 3. The method according to claim 1 or 2,
The composition is characterized in that to reduce the activity of alcohol dehydrogenase (ADH) or reduce the activity of aldehyde dehydrogenase (ALDH), liver function improvement or hangover pharmaceutical composition. Chlorella extract; And
Health functional food for improving liver function or hangover, comprising herbal extracts consisting of pine needles, mulberry leaves, barley sprouts, pumpkins, thorny cucumbers, mugwort, eoseongcho, fresh vinegar, motherwort, dandelion and cornus as active ingredients. Dry powder of chlorella; And
Health functional food for improving liver function or hangover, comprising herbal dry powder consisting of pine needles, mulberry leaves, barley shoots, pumpkin, thorny greens, mugwort, eoseongcho, fresh vinegar, motherwort, dandelion and cornus as active ingredients.