KR20210109074A - Food Composition Comprising Extract of Paullinia cupana, Vitamin and Natural Extract as an Active Ingredients - Google Patents

Abstract

The present invention relates to a food composition comprising a Paullinia cupana extract, vitamins, and a natural substance extract as active ingredients. When the food composition of the present invention is used, effects of fatigue recovery, anti-oxidation, and DNA damage protection can be exhibited.

Description

Food composition comprising guarana extract, vitamin and natural extract as active ingredients {Food Composition Comprising Extract of Paullinia cupana, Vitamin and Natural Extract as an Active Ingredients}

The present invention relates to a food composition comprising a guarana extract, a vitamin and a natural product extract as an active ingredient.

As the emergence of modern well-being culture represents, domestic consumers' interest in health has increased significantly compared to the past. In addition, as advances in science bring extension of human lifespan, health functional food has emerged as a breakthrough in the future food industry.

The global functional food market, which was worth 65 billion dollars in 1997, showed a high growth rate to 138 billion dollars in 2000, and the domestic health functional food market also contracted temporarily during the 1997 Asian financial crisis. there is a trend

Energy drink, one of health functional foods, refers to a functional drink that helps recover from fatigue by replenishing moisture and electrolytes lost from the body due to exercise or labor. of minerals and also contains vitamin C.

Energy drinks contain taurine, caffeine, and nutrients (vitamin C, amino acids, etc.) to strengthen concentration and increase the vitality of the body. However, since caffeine is the main ingredient that increases the vitality of the body, there is a problem that sleep can be disturbed if you drink it in the evening. Also, consumers have a general preconceived notion that energy drinks are unhealthy.

Therefore, there is a need to develop an energy drink with improved functionality to overcome the problems of the existing energy drink.

Republic of Korea Patent No. 10-2045192

The present inventors made intensive research efforts to develop an energy drink with improved functionality. As a result, in the case of a food composition comprising a guarana extract, a vitamin and a natural extract, the present invention was completed by finding that it exhibits a fatigue recovery effect, an antioxidant effect, and a DNA damage protection effect.

Accordingly, an object of the present invention is to provide a food composition comprising a guarana extract, a vitamin and a natural product extract as an active ingredient.

According to one aspect of the present invention, the present invention provides a food composition comprising a guarana extract, a vitamin and a natural extract as an active ingredient.

Guarana (scientific name: Paullinia cupana) or Guarana is a vine of the genus Polynia, native to the Amazon Basin of Brazil. Guarana is a fruit of a plant that promotes recovery from fatigue, blood circulation, and brain activity. It contains about three times as much caffeine as coffee. characteristic.

As used herein, the term "extract" has the meaning commonly used as a crude extract in the art, but in a broad sense also includes a fraction obtained by additionally fractionating the extract. That is, the guarana extract or natural product extract includes not only those obtained using an extraction solvent, but also those obtained by additionally applying a purification process thereto. For example, a fraction obtained by passing the extract through an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (prepared for separation according to size, charge, hydrophobicity or affinity), etc. The fraction obtained through the purification method is also included in the extract of the present invention.

When the extract used in the composition of the present invention is obtained by treating Guaranae with an extraction solvent, various extraction solvents may be used. Preferably, a polar solvent or a non-polar solvent may be used. Suitable as polar solvents are: (i) water, (ii) alcohols (preferably methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, n-butanol), (iii) acetic acid, (iv) DMFO (dimethyl-formamide) and (v) DMSO (dimethyl sulfoxide).

Suitable nonpolar solvents are acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, hexane, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1- Pentene, 1-chlorobutane, 1-chloropentane, o-xylene, diisopropyl ether, 2-chloropropane, toluene, 1-chloropropane, chlorobenzene, benzene, diethyl ether, diethyl sulfide, chloroform, dichloro methane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride and THF.

In the present invention, when extracting guarana, hot water extraction method, cold extraction method, reflux extraction method, room temperature extraction method, or ultrasonic extraction method may be used, but is not limited thereto.

In one embodiment of the present invention, the guarana extract is extracted with any one extraction solvent selected from the group consisting of water, an organic solvent, and a mixed solvent thereof, and the organic solvent is alcohol, petroleum ether, hexane, benzene, It is selected from the group consisting of chloroform, methylene chloride, ether, ethyl acetate, and acetone.

In one embodiment of the present invention, the alcohol is selected from the group consisting of methanol, ethanol, propanol, butanol, normal-propanol, iso-propanol, normal-butanol, and a mixed solvent thereof.

In one embodiment of the present invention, the concentration of the ethanol solution is 30 to 100% by volume.

More specifically, the concentration of the ethanol solution is 50 to 99% by volume, 60 to 99% by volume, 70 to 99% by volume, 80 to 99% by volume, 90 to 99% by volume, 50 to 98% by volume, 60 to 98% by volume , 70 to 98% by volume, 80 to 98% by volume, 90 to 98% by volume. Most specifically, the concentration of the ethanol solution is 95 to 98% by volume.

Guarana extract used in the present invention may be prepared in a powder state by an additional process such as distillation under reduced pressure and freeze-drying or spray-drying.

As used herein, the term "natural extract" is a compound or substance produced by a living organism found in nature, and the natural extract may be extracted from a culture medium of land plants, marine organisms, or microorganisms.

Since the content regarding the extraction method of the natural extract contained in the food composition of the present invention is the same as the content regarding the extraction method of the guarana extract, the content common between the two is to avoid excessive complexity of the present specification, the description omit

As used herein, the term "comprising as an active ingredient" means including an amount sufficient to achieve the efficacy or activity of the guarana extract, vitamins, and natural extracts.

Guarana extract and natural product extract contained in the composition of the present invention is a natural material and has no cytotoxicity. The upper limit of the quantity of extracts and vitamins contained in the composition of the present invention may be selected by those skilled in the art within an appropriate range.

The food composition of the present invention may include not only the extract and vitamins as active ingredients, but also ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents. include Examples of the above-mentioned carbohydrates include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose, oligosaccharides and the like; and polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [taumatine, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is prepared as a drink, citric acid, high fructose, sugar, glucose, acetic acid, malic acid, fruit juice, etc. may be additionally included in addition to the guarana extract of the present invention.

In one embodiment of the present invention, the guarana extract, vitamins and natural extracts are mixed in a weight ratio of 1:0.1-0.9:0.1-0.9.

More specifically, the weight ratio of the guarana extract, vitamins and natural extracts is 1:0.1-0.7:0.1-0.9, 1:0.1-0.5:0.1-0.9, 1:0.1-0.4:0.1-0.9, 1:0.2-0.9 :0.1-0.9, 1:0.2-0.7:0.1-0.9, 1:0.2-0.5:0.1-0.9, 1:0.1-0.9:0.2-0.7, 1:0.1-0.9:0.2-0.5, 1:0.1-0.9 :0.3-0.7, 1:0.1-0.9:0.3-0.5, 1:0.2-0.7:0.2-0.7, 1:0.2-0.7:0.2-0.5, 1:0.2-0.5:0.2-0.7, 1:0.2-0.5 :0.2-0.5, 1:0.2-0.5:0.3-0.7, 1:0.2-0.5:0.3-0.5, 1:0.2-0.5:0.4-0.7, 1:0.2-0.5:0.4-0.6. Most specifically, it is mixed in a weight ratio of 1:0.2-0.4:0.4-0.5.

In one embodiment of the present invention, the vitamin is selected from the group consisting of vitamin A, vitamin B group, vitamin C, vitamin D, vitamin E, vitamin K, and combinations thereof.

More specifically, the vitamin is selected from the group consisting of vitamin B group, vitamin C, vitamin E, and combinations thereof.

In one embodiment of the present invention, the vitamin B group is vitamin B ₁ , vitamin B ₂ , vitamin B ₃ , vitamin B ₄ , vitamin B ₅ , vitamin B ₆ , vitamin B ₇ , vitamin B ₉ , vitamin B ₁₂ , choline and combinations thereof.

More specifically, the vitamin B group is _{selected from the group consisting of vitamin B 1} , vitamin B ₂ , vitamin B ₅ , choline, and combinations thereof.

In one embodiment of the present invention, the natural product is Schisandra, melon, flounder, peach, gold kiwi, strawberry, cherry tomato, songbon, melon, red kiwi, green grape, jujube, black tomato, dandelion fruit, maru fruit, geobong, apple , Red glove, Aori apple, pear, tangerine, garlic, red beet, cucumber pepper, water parsley, pear, zucchini, cucumber, purple potato, onion, grape, spinach, purple sweet potato, carrot, zucchini pumpkin, apricot, green pepper, broccoli, king oyster It is selected from the group consisting of mushrooms, sweet potatoes, watermelons, Korean pumpkins, and combinations thereof.

More specifically, the natural product is selected from the group consisting of tangerines, sweet persimmons, strawberries, melons, apples, omija, tomatoes, and combinations thereof.

Tangerine is a plant of the genus citrus that corresponds to the citrus family, and tangerine, hallabong, cheonhyehyang, cheongyeon, kumquat, etc. may correspond to this. Mandarin oranges are rich in vitamin C and beta-carotene, which strengthens immunity and aids in eye health.

Sweet persimmon (Diospyros kaki) is rich in vitamins C and A, so it is not only good for preventing colds, especially during the changing seasons, but also useful for preventing infectious diseases and physiological activities of the eyes. In addition, persimmons contain protein, fat, carbohydrates, ash, phosphoric acid and iron together.

Strawberry is a perennial plant belonging to the Rosaceae family and belongs to a crop that has a long cultivation period and requires a lot of labor, but can be grown at low temperatures and can be harvested from November to May through low-temperature facility cultivation. Flavonoids in cultivated strawberries include quercetin-7-xyloside, fragarin, procyanidin B3, 1-Op-coumaroylglucose ), 2-heptanol (2-heptanol) or ellagic acid is known to include. Therefore, it is effective in diseases such as anti-inflammatory, anti-cancer, and kidney disease using strawberries.

Melon has a high sugar content and contains a lot of iron, niacin, vitamins A and C. It is an anticoagulant that lowers blood viscosity and prevents heart disease or stroke. .

Apples are the fruits of the apple tree, a plant of the deciduous tree of the Rosaceae, a dicotyledonous plant, and contain a large amount of dietary fiber, vitamins, and minerals in addition to palatable ingredients such as organic acids and sugars. have.

Omija is the fruit of the Schisandra tree, and the Schisandra tree is a deciduous broad-leaved vine that grows widely in Mt. Jiri, Mt. Songni, and Mt. Taebaek. Omija has a strong sour taste and ripens in red from August to September. Omija contains ingredients such as shizandrine, gomisin, citral, malic acid, and citric acid, and is used as a tonic by strengthening the heart, lowering blood pressure, and increasing immunity. In addition, omija strengthens lung function and has antitussive and expectorant effects, so it is effective in treating cough and thirst.

Tomato (Solanum lycopersicum) has been reported that lycopene inhibits aging by preventing cell oxidation, and has a preventive effect on cardiovascular disease and various cancers. It contains 8 times more vitamin A than strawberries, and vitamin C contains the highest amount among vegetable crops. As for the organic acid content of tomatoes, citric acid accounts for 0.5 to 21%, and free amino acids 0.07 to 0.09% to give a unique taste, prevent aging, suppress degenerative diseases, and strengthen human immune function. Due to its ingredients, it strengthens blood vessels and lowers blood pressure.

In one embodiment of the present invention, the food composition of the present invention is to include a guarana extract, a vitamin and a tangerine extract, compared to 100 parts by weight of the guarana extract, the weight part of the tangerine extract is 1 to 30 parts by weight. More specifically, compared to 100 parts by weight of the guarana extract, the parts by weight of the tangerine extract are 3 to 25, 5 to 20, 7 to 15 parts by weight. Most specifically, compared to 100 parts by weight of the guarana extract, the weight part of the tangerine extract is 10 to 15 parts by weight.

In one embodiment of the present invention, the food composition of the present invention is to include a guarana extract, a vitamin and a strawberry extract, compared to 100 parts by weight of the guarana extract, the weight part of the strawberry extract is 1 to 30 parts by weight. More specifically, relative to 100 parts by weight of the guarana extract, parts by weight of the strawberry extract are 3 to 25, 5 to 20, and 7 to 15 parts by weight. Most specifically, relative to 100 parts by weight of the guarana extract, the weight part of the strawberry extract is 10 to 15 parts by weight.

In one embodiment of the present invention, the food composition of the present invention includes a guarana extract, a vitamin, and a Schisandra extract, and 100 parts by weight of the guarana extract, the weight part of the Schisandra extract is 1 to 30 parts by weight. More specifically, compared to 100 parts by weight of the Guarana extract, the weight part of the Schisandra extract is 3 to 25, 5 to 20, 7 to 15 parts by weight. Most specifically, compared to 100 parts by weight of the Guarana extract, the weight part of the Schisandra extract is 7 to 13 parts by weight.

In one embodiment of the present invention, the food composition of the present invention comprises a guarana extract, a vitamin and a sweet persimmon extract, compared to 100 parts by weight of the guarana extract, the weight part of the sweet persimmon extract is 1 to 10 parts by weight. More specifically, compared to 100 parts by weight of the guarana extract, the weight part of the sweet persimmon extract is 3 to 9, 4 to 8 parts by weight. Most specifically, compared to 100 parts by weight of the guarana extract, the weight part of the sweet persimmon extract is 5 to 7 parts by weight.

In one embodiment of the present invention, the food composition of the present invention is to include a guarana extract, vitamins and tomato extract, compared to 100 parts by weight of the guarana extract, the weight part of the tomato extract is 0.5 to 8 parts by weight. More specifically, compared to 100 parts by weight of the guarana extract, parts by weight of the tomato extract are 1 to 6 parts by weight, 1.5 to 4.5 parts by weight. Most specifically, compared to 100 parts by weight of the guarana extract, parts by weight of the tomato extract are 2 to 4 parts by weight.

In one embodiment of the present invention, the food composition of the present invention is to include a guarana extract, vitamin C and a natural extract, compared to 100 parts by weight of the guarana extract, the weight part of the vitamin C is 0.5 to 8 parts by weight. More specifically, compared to 100 parts by weight of the guarana extract, the weight part of the vitamin C is 1 to 6, 1.5 to 4.5 parts by weight. Most specifically, compared to 100 parts by weight of the guarana extract, the weight part of the vitamin C is 2 to 4 parts by weight.

In one embodiment of the present invention, the food composition of the present invention is to include a guarana extract, vitamin E and a natural extract, compared to 100 parts by weight of the guarana extract, the weight part of the vitamin E is 0.05 to 0.5 parts by weight. More specifically, relative to 100 parts by weight of the guarana extract, the weight part of the vitamin E is 0.1 to 0.3 parts by weight.

In one embodiment of the present invention, the food composition of the present invention is to include guarana extract, vitamin B group and natural extract, compared to 100 parts by weight of the guarana extract, the weight part of the vitamin B group is 1 to 60 parts by weight . More specifically, compared to 100 parts by weight of the guarana extract, the parts by weight of the vitamin B group are 5 to 50, 10 to 45, 15 to 40 parts by weight. Most specifically, compared to 100 parts by weight of the guarana extract, the weight part of the vitamin B group is 15 to 35 parts by weight.

In one embodiment of the present invention, the food composition of the present invention is to include a guarana extract, choline and a natural extract, compared to 100 parts by weight of the guarana extract, the weight part of the choline is 5 to 50 parts by weight. More specifically, compared to 100 parts by weight of the guarana extract, the weight part of the choline is 10 to 40, 15 to 35, 15 to 30 parts by weight. Most specifically, compared to 100 parts by weight of the guarana extract, the weight part of the choline is 18 to 26 parts by weight.

In one embodiment of the present invention, the food composition of the present invention is to include a guarana extract, vitamin B ₁ , vitamin B ₂ , vitamin B _{5 and a natural extract, and the vitamin B 1} , vitamins compared to 100 parts by weight of the guarana extract B ₂ and vitamin B ₅ are in an amount of 0.05 to 1 part by weight. More specifically, the amount of vitamin B ₁ , vitamin B ₂ , and vitamin B ₅ based on 100 parts by weight of the guarana extract is 0.1 to 0.9, 0.2 to 0.8, 0.3 to 0.8 parts by weight. _{Most specifically, the amount of vitamin B 1} , vitamin B ₂ , and vitamin B ₅ is 0.4 to 0.8 parts by weight relative to 100 parts by weight of the guarana extract.

In one embodiment of the present invention, the food composition of the present invention comprises a guarana extract, vitamin B ₃ and a natural extract, and the weight part of the vitamin B ₃ relative to 100 parts by weight of the guarana extract is 0.1 to 5 parts by weight. _{More specifically, the amount of vitamin B 3} relative to 100 parts by weight of the guarana extract is 0.2 to 4, 0.3 to 3, 0.4 to 2 parts by weight. Most specifically, the weight part of the vitamin B ₃ relative to 100 parts by weight of the guarana extract is 0.5 to 1.5 parts by weight.

In one embodiment of the present invention, the food composition of the present invention comprises a guarana extract, vitamin B ₆ and a natural extract, and the weight part of the vitamin B ₆ relative to 100 parts by weight of the guarana extract is 0.01 to 0.2 parts by weight. More specifically, the weight part of the vitamin B ₆ relative to 100 parts by weight of the guarana extract is 0.03 to 0.17 parts by weight. Most specifically, the weight part of the vitamin B ₆ relative to 100 parts by weight of the guarana extract is 0.05 to 0.15 parts by weight.

In one embodiment of the present invention, the food composition of the present invention comprises a guarana extract, vitamin B ₁₂ and a natural extract, and the weight part of the vitamin B ₁₂ relative to 100 parts by weight of the guarana extract is 0.0005 to 0.0015 parts by weight. More specifically, the weight part of the vitamin B ₁₂ relative to 100 parts by weight of the guarana extract is 0.0007 to 0.0014 parts by weight.

In one embodiment of the present invention, the food composition is a composition for recovery from fatigue.

In one embodiment of the present invention, the food composition is an antioxidant composition.

In one embodiment of the present invention, the food composition is a composition for protecting DNA damage.

The DNA damage of the present invention is induced by oxidative stress, but is not limited thereto.

In one embodiment of the present invention, the form of the composition is selected from the group consisting of powders, granules, tablets, capsules, and beverages.

More specifically, the composition is in the form of a powder.

According to an embodiment of the present invention, a food composition comprising a guarana extract, a vitamin, and a natural product extract _{has a recovery effect on oxidative stress induced by H 2} O ₂ , and it was confirmed that the total flavonoid content was high. In addition, it was confirmed that the composition of the present invention has an excellent antioxidant effect through DPPH radical scavenging activity analysis, ORAC activity analysis, and CAC analysis.

According to an embodiment of the present invention, it was confirmed that the composition of the present invention had an effect of protecting DNA damage due to oxidative damage through comet assay, and it was confirmed that there was a fatigue recovery effect through a survey.

In conclusion, the food composition comprising the guarana extract, vitamins, and natural extracts of the present invention has an antioxidant effect, a DNA damage protection effect, and a fatigue recovery effect.

The features and advantages of the present invention are summarized as follows:

The present invention provides a food composition comprising a guarana extract, a vitamin and a natural extract as an active ingredient.

When the food composition of the present invention is used, it exhibits a fatigue recovery effect, an antioxidant effect, and a DNA damage protection effect.

1 shows the results of comparing the oxidative stress recovery effect of an energy drink mix measured by MTT assay with other commercially available products.
Figure 2 shows the results of comparing the antioxidant effect of the energy drink mix analyzed by measuring the total polyphenol content through the Folin-Denis method with other commercially available products.
3 shows the results of comparing the antioxidant effect of the energy drink mix analyzed by measuring the total flavonoid content through the Davis method with other commercially available products.
4 shows the results of comparing the antioxidant effect of the energy drink mix analyzed by measuring the DPPH radical scavenging ability with other commercially available products.
5 shows the results of comparing the antioxidant effect of the energy drink mix analyzed using the ORAC method with other commercially available products.
6 shows the results of comparing the antioxidant effect of the energy drink mix analyzed by measuring CAC (Cellular antioxidant capacity) with other commercially available products.
7 shows the results of comparing the DNA damage protection effect of the energy drink mix analyzed by measuring the content of tail DNA through the Comet assay with other commercially available products.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

Example

Throughout this specification, "%" used to indicate the concentration of a specific substance is (weight/weight) % for solid/solid, (weight/volume) % for solid/liquid, and Liquid/liquid is (volume/volume) %.

Preparation Example 1: Preparation of guarana extract used for energy drink mix preparation

Brazilian Guarana fruit extract powder was purchased (MOS Mall, Seoul, Korea) and used while stored in a refrigerator at 5 °C. The extracted powder was extracted with 95% ethanol, and the concentration was concentrated at 37°C using a rotary vacuum concentrator (EYELA N-1000, Tokyo Rikakikai Co., Tokyo, Japan).

Preparation Example 2: Preparation of natural powder used for energy drink mix preparation

The natural products to be used in the energy drink mix production are tangerines, sweet persimmons, strawberries, melons, apples, omija, and tomatoes. After freeze-drying the natural product, it was pulverized by pulverizing with a grinder, and the pulverized freeze-dried powder was used while being stored in a refrigerator at 5°C. The natural product was extracted with 95% ethanol, and the concentration was concentrated at 37°C with a rotary vacuum concentrator (EYELA N-1000, Tokyo Rikakikai Co., Tokyo, Japan).

Preparation Example 3: Preparation of energy drink mix

All materials were powdered to prepare the energy drink mix, and guarana, vitamins, and natural ingredients, which are the main ingredients of the energy drink mix, were prepared in the composition shown in Table 1.

ingredient content Guarana Extract 3 g vitamin B _One 3 mg vitamin B ₂ 3 mg Niacin (Vitamin B) ₃ ) 34 mgNE Pantothenic acid (vitamin B) ₅ ) 12 mg vitamin B ₆ 4 mg vitamin B ₁₂ 34 μg vitamin C 100 mg vitamin E 6 mga-TE choline 678.2 mg tangerine extract 0.4 g strawberry extract 0.4 g Schisandra extract 0.3 g sweet persimmon extract 0.2 g tomato extract 0.1 g

Example 1: Oxidative stress recovery effect of energy drink mix

Mouse-origin myocyte (C2C12) cell line (ATCC, Virginia, USA) was added to DMEM medium (Sigma Aldrich, Missouri, USA) with 10% fetal bovine serum (FBS), 1% glucose, 1% glutamine ( glutamine), 100 U/mL penicillin, and 100 μg/mL streptomycin in a culture medium containing 95% humidity at 37 °C, 5% CO ₂ Incubated in an incubator.

C2C12 cells were seeded in a 96-well plate at a concentration of ^{1×10 5} cells/mL the day before the experiment _{, treated with the sample and 1 mM H 2} O ₂ , and incubated for 24 hours. Thereafter, MTT stored at 4°C was dissolved in PBS at a concentration of 5 mg/mL and then added using a syringe, and an amount equal to 20% of the volume of 96-well medium was taken using a pipette to each well. was carefully added to the mixture, and the plate was attached to the floor by gently shaking back and forth and left and right. After culturing for 4 hours in a CO ₂ incubator, the culture solution was removed, and formazan was eluted by adding 100 μL of DMSO, followed by _{incubation in a CO 2} incubator for 30 minutes. Cell viability was calculated by measuring at 540 nm absorbance after 30 min.

As a result of the MTT reduction analysis performed to measure the oxidative stress recovery effect, the cell viability (26.3 ± 1.3%) of the energy drink mix of Preparation Example 3 was higher than that of PC (positive control group) induced by oxidative stress with _{1 mM H 2} O ₂ significantly as high as 23.5% (Table 2, Figure 1).

division NC PC A B C cell
survival rate 99.9 ± 1.2 ^a 21.3 ±0.4 ^c 28.7 ± 1.7 ^b 26.4 ± 26.3 ^b 26.3 ± 1.3 ^b

NC: normal control, PC: _{positive control induced by oxidative stress with 1 mM H 2} O ₂ A: Commercial products A, B: Commercial products B, C: Energy drink mix of Preparation Example 3

a-c; Significance for each group was indicated (p<0.05)

Example 2: Analysis of antioxidant effect of energy drink mix

Example 2-1. Analysis of total polyphenol content in energy drink mixes

The total polyphenol content was colorimetrically quantified according to the Folin-Denis method. Add 1 mL of distilled water to 1 mL of sample to dilute, add 2 mL of 1 N Folin-Ciocalteu's phenol reagent, leave at room temperature for 3 minutes, add 2 mL of 10% Na ₂ CO ₃ solution, and leave the mixture at room temperature for 1 hour. did. After 1 hour, the mixture was centrifuged at 13,400 g for 5 minutes, 200 μL of the supernatant was taken, and absorbance was measured at 690 nm using an ELISA reader (Sunrise, Tecan Co. Ltd., Grφdig, Austria). A calibration curve was prepared using gallic acid as a standard material, and the total phenol content was expressed in mg GAE/100 g (mg garlic acid equivalents/100 g).

The total polyphenol content of the energy drink mix of Preparation Example 3 was 300.5 ± 1.3 mg GAE/100 g, which was significantly lower than that of commercial products A and B (Table 3, FIG. 2 ).

division A B C Total polyphenols (mg GAE/100 g) 1496.1 ± 3.0 ^c 723.4 ± 1.7 ^b 300.5 ± 1.3 ^a

A: Commercial products A, B: Commercial products B, C: Energy drink mix of Preparation Example 3

a-c; Significance for each group was indicated (p<0.05)

Example 2-2. Analysis of total flavonoid content of energy drink mixes

Total flavonoid content was colorimetrically quantified according to the Davis method. 1 mL of diethyl glycol was added to 0.1 mL of the sample, left at room temperature for 5 minutes, 0.1 mL of 1 N NaOH solution was added, and the mixture was left at room temperature for 30 minutes. The absorbance of the reaction mixture was measured at 420 nm using a UV/VIS spectrometer-(UV 1601, shimadzu, Kyoto, Japan). A calibration curve was prepared using quercetin as a standard material, and the total flavonoid content was expressed in mg QE/100 g (mg quercetin equivalents/100 g).

The total flavonoid content of the energy drink mix of Preparation Example 3 was 1076.8 ± 0.0 mg QE/100 g, which was 127.1% of the commercial product A (474.2 ± 1.6 QE/100 g) and the commercial product B (298.8 ± 0.4 QE/100 g), respectively. , was significantly higher by 260.4% (Table 4, Fig. 3).

division A B C Total Flavonoids (mg QE/100 g) 474.2 ± 1.6 ^b 298.8 ± 0.4 ^a 1076.8 ± 0.0 ^c

A: Commercial products A, B: Commercial products B, C: Energy drink mix of Preparation Example 3

a-c; Significance for each group was indicated (p<0.05)

Example 2-3. Analysis of DPPH radical scavenging activity of energy drink mix

DPPH radical scavenging ability was measured by modifying the method of Mensor et al. Add 80 μL of 0.4 mM DPPH ethanol solution to 20 μL of the sample, mix well for 10 seconds, and then react at room temperature for 10 minutes. Absorbance was measured at 492 nm.

The DPPH radical scavenging ability of the energy drink mix of Preparation Example 3 was 65.7 ± 0.7%, indicating the same activity as that of commercial product B (64.2 ± 0.6%), and was significantly higher than that of commercial product A (25.1 ± 3.3%) by 161.8%. (Table 5, Figure 4).

division A B C DPPH radical scavenging ability (%) 25.1 ± 3.3 ^a 64.2 ± 0.6 ^b 65.7 ± 0.7 ^b

A: Commercial products A, B: Commercial products B, C: Energy drink mix of Preparation Example 3

a-b; Significance for each group was indicated (p<0.05)

Example 2-4. Oxygen radical absorbance capacity (ORAC) activity analysis of energy drink mixes

Antioxidant activity was measured through peroxyl radical scavenging ability using peroxyl radical scavenging capacity (ORAC _ROO ) analysis. 100 μL of buffer, 50 μL of 80 nM AAPH for generation of peroxyl radicals, and 50 μL of 80 nM fluorescein were added to 50 μL of sample, followed by a GENios fluorescence plate reader (Tecan Trading AG, Salzburg, Austria). It was measured for 2 hours every 2 minutes at an excitation wavelength of 485 nm and an emission wavelength of 535 nm using the The final reaction concentration of 1 μM trolox was used as a standard, and the ORAC value was calculated as the net area under the curve in which the fluorescence of each sample decreased, and 1 μM Trolox equivalent ( trolox equivalents, TE).

The ORAC activity of the energy drink mix of Preparation Example 3 was 3.86 ± 0.05 1 µM TE at a concentration of 50 µg/mL, and there was no significant difference from the commercial product A (3.51 ± 0.28 %), but was as high as 10.0 %, and the commercial product B (3.02 ± 0.29%) by 27.8% (Table 6, Fig. 5).

division A B C ORAC _ROO (1 μM TE) 3.51 ± 0.28 ^ab 3.02 ± 0.29 ^a 3.86 ± 0.05 ^b

A: Commercial products A, B: Commercial products B, C: Energy drink mix of Preparation Example 3

a-b; Significance for each group was indicated (p<0.05)

Example 2-5. Cellular antioxidant capacity (CAC) analysis of energy drink mixes

Human-origin hepatocyte (HepG2) cell line (KCLB, Seoul, Korea) was added to DMEM medium (Sigma Aldrich, Missouri, USA) with 10% fetal bovine serum (FBS), 1% glucose, 1% glutamine, 100 U/mL penicillin, 100 It was cultured in a culture medium containing μg/mL streptomycin at 37° C., maintained at 95% humidity, in an incubator of _{5% CO 2 .}

HepG2 cells were counted at 5×10 ⁵ cells/mL, and 100 μL of each was added to a 96-well plate (Greiner, 655090) and cultured in an incubator at _{37° C., 5% CO 2 .} After 24 hours, the medium was removed, 200 µL of fluorescence-stable Hank's balanced salt solution (HBSS) was dispensed into each well, and 2 µL of each sample was treated to a final concentration of 100 µg/mL and incubated for 30 minutes. Each well was washed with 50 μL of HBSS, then 200 μL of HBSS was again dispensed, and 2 μL of 8 mM AAPH was added with a peroxyl radical generator and incubated for 30 minutes. At this time, AAPH was prepared fresh each time and used. After adding 2 μL of 40 mM DCFH-DA as a fluorescent probe, blocking the light and incubating for 30 minutes, fluorescence using a FLUOstar OPTIMA microplate reader (BMG Lab Technology; 485 nm excitation wavelength and 538 nm emission wavelength) was measured. As a control group, only DCFH-DA was treated, and AAPH and DCFH-DA were treated as a positive control. The relative fluorescence values of each sample and the positive control, AAPH, were compared by setting the fluorescence value of the control to 100%.

The CAC activity of all products was significantly reduced compared to PC (positive control), and the CAC activity of the energy drink mix was 127.2 ± 1.4%, which was 70.1% lower than that of PC, showing a high effect of protecting cells from AAPH radicals. In addition, the CAC activity of the energy drink mix was 25.8% and 16.6% higher than those of commercial product A and commercial product B, respectively (Table 7, FIG. 6).

division NC PC A B C DCF fluorescence intensity 100.0 ± 4.5 ^a 197.3 ± 28.8 ^d 160.0 ± 5.0 ^c 148.3 ± 0.7 ^bc 127.2 ± 1.4 ^b

NC: normal control group, PC: positive control group induced by oxidative stress with AAPH, A: commercial product A, B: commercial product B, C: energy drink mix of Preparation Example 3

a-d; Significance for each group was indicated (p<0.05)

Example 3: Analysis of DNA damage protection effect of energy drink mix

Human-derived hepatocyte (HepG2) cell line is cultured in DMEM medium containing 10% fetal bovine serum (FBS), 1% glucose, 1% glutamine, 100 U/mL penicillin, and 100 µg/mL streptomycin, maintaining 95% humidity. 37 ° C., 5% CO ₂ Incubated in an incubator.

In order to induce artificial oxidative stress after treating the separated hepatocytes with samples by concentration (1, 5, 10, 50 μg/mL) and reacting at 37 °C for 30 minutes, after the reaction is completed, hepatocytes are washed with PBS. 200 μM of H ₂ O ₂ was treated at 4° C. for 5 minutes, and then washed with PBS. After centrifugation at 500 g for 5 minutes, the obtained cells are mixed evenly in 0.7% low melting agarose (LMA), and then placed on a slide precoated with 1% normal melting agarose (NMA) and placed on a cover glass. Covered and evenly dispersed, then left at 4 °C for 20 minutes. When the gel hardened, the cover glass was removed, layered again with 75 µL of 0.7% LMA solution, and left at 4 °C for 20 minutes. When the gel hardens, immerse the slides in cold alkaline lysis buffer (2.5M NaCl, 100 mM Na ₂ EDTA, 10 mM Tris, 1% sodium lauryl sarcosine, 1% Triton X-100, 10% DMSO) in the dark at 4 °C. DNA double strands were released by immersion for 1 hour. After dissolution, the slides were left in an electrophoresis buffer (300 mM NaOH, 10 mM Na ₂ EDTA, pH>13) at 4 ° C for 20 minutes to release alkali labile sites of DNA. Electrophoresis was performed for 20 minutes at a voltage of 25 V/300±3 mA. After electrophoresis, the slides were washed three times for 5 minutes in cold neutral solution (0.4 M Tris buffer, pH 7.4) and once in ethanol for 5 minutes, and then dried. The slides were stained with ethidium bromide (20 μg/mL), and the DNA damage level was measured using a CCD camera (Nikon, Tokyo, Japan) of a fluorescence microscope (LEICA DMLB, Wetzlar, Germany). (Komet version 5.0, Kinetic Imaging, Liverpool, UK) was used for analysis.

As a result of the comet assay performed to measure the DNA damage protection effect, the energy drink mix of Preparation Example 3 significantly reduced tail DNA (%) by 75.2% compared to PC (positive control). In addition, the energy drink mix of Preparation Example 3 showed the effect of reducing the tail DNA (%) by 46.0% than the commercial product A and by 29.4% than the commercial product B.

division NC PC A B C Head DNA (%) 96.0 ± 0.1 ^c 17.9 ± 6.9 ^a 62.2 ± 15.7 ^b 71.1 ± 4.7 ^b 79.6 ± 4.7 ^b Tail DNA (%) 4.0 ± 0.1 ^a 82.1 ± 6.9 ^c 37.8 ± 15.7 ^b 28.9 ± 4.7 ^b 20.4 ± 4.7 ^b Tail extent moment 5.0 ± 0.7 ^a 385.5 ± 65.8 ^b 128.2 ± 65.7 ^a 104.1 ± 23.9 ^a 68.1 ± 39.0 ^a Tail Length 107.1 ± 15.8 ^a 450.9 ± 56.4 ^b 178.2 ± 83.6 ^a 159.5 ± 34.9 ^a 88.8 ± 39.9 ^a

NC: normal control, PC: _{positive control induced by oxidative stress with 1 mM H 2} O ₂ A: Commercial products A, B: Commercial products B, C: Energy drink mix of Preparation Example 3

a-c; Significance for each group was indicated (p<0.05)

Example 4: Analysis of anti-fatigue effect of energy drink mix through survey

All beverages and foods containing caffeine were prohibited from 6 hours before the study (14:00), and the same dinner was provided to the subjects at 18:00. The intake time of the energy drink mix and the placebo drink mix was 22:00.

Subjects were asked to 'fatigue items' 6 times before (21:30~21:50, once), immediately after ingestion (22:00), and at 4:00 a.m. the next day after ingestion of the energy drink mix and placebo drink mix at 1 hour intervals. A questionnaire (a list of symptoms of fatigue perception suggested by the Industrial Fatigue Committee of the Japan Industrial Hygiene Association in 1967 and confirmed by the association in 1970 (published by the Japan Labor Science Research Institute, 1970)) was prepared. After 4:00 am, each took a break and completed the 'Fatigue Questionnaire' and 'Effect of Energy Drink Mix Intake Questionnaire' at 7:00 am.

If drowsiness of 5 minutes or less occurred before 4:00 am, the study time, the study was continued. 'Item Questionnaire' and 'Energy Drink Mix Intake Effect Item Questionnaire' were completed. After a week-long drug-free period, the same experiment as above was performed by cross-ingesting an energy drink mix or a placebo drink mix.

The subjective symptoms of fatigue were divided into physical symptoms, mental symptoms, and neurosensory symptoms, and each item consisted of 10 items according to the symptoms. 0 points were given if symptoms were present and 1 point was given if there were no symptoms.

As a result of conducting a survey on the symptoms of fatigue after consuming an energy drink mix, the physical symptoms showed that 1 hour (9.00 ± 1.41), 2 hours (8.33 ± 2.55), 5 hours (8.33 ± 2.55), and 5 hours (8.33 ± 2.55), After 9 hours (8.22 ± 1.79), the anti-fatigue effect was significantly higher than that of the placebo group. In neurosensory symptoms, the anti-fatigue effect was significantly higher than that of the placebo group at 1 hour (10.00 ± 0.00), 2 hours (10.00 ± 0.00), and 3 hours (9.78 ± 0.67) after ingestion of the energy drink mix (Table 9). ).

When the fatigue analysis result of the energy drink mix group 1 hour after ingestion and 2 hours after ingestion, which had the highest effect, was expressed on a 7-point scale, in the physical symptom category, 1.5 points after 1 hour of ingestion and 2.3 points after 2 hours after ingestion were obtained, and In the symptom item, 1 point after 1 hour of ingestion and 1 point after 2 hours of ingestion were indicated.

after ingestion physical symptoms mental symptoms neurosensory symptoms placebo group energy drink mix placebo group energy drink mix placebo group energy drink mix 0 H 8.11 ± 1.90 8.11 ± 1.76 9.44 ± 1.01 9.11 ± 1.54 9.44 ± 0.88 9.67 ± 0.71 1 H 7.22 ± 1.79 9.00 ± 1.41* 9.00 ± 1.66 9.78 ± 0.44 8.78 ± 1.20 10.00 ± 0.00* 2H 5.78 ± 2.05 8.33 ± 2.55* 8.33 ± 2.29 9.67 ± 0.71 8.78 ± 1.56 10.00 ± 0.00* 3H 5.33 ± 2.24 6.56 ± 2.70 8.33 ± 2.18 8.44 ± 1.42 8.44 ± 1.51 9.78 ± 0.67* 4H 5.13 ± 2.10 6.88 ± 2.53 7.00 ± 3.16 8.38 ± 1.51 8.63 ± 1.51 9.13 ± 1.13 5H 3.00 ± 2.27 5.50 ± 3.12* 6.75 ± 3.06 7.63 ± 2.62 7.75 ± 1.98 8.50 ± 2.33 6H 3.13 ± 2.03 4.75 ± 3.06 6.25 ± 3.01 7.25 ± 3.45 6.75 ± 2.25 8.38 ± 2.26 9H 5.22 ± 3.27 8.22 ± 1.79* 7.22 ± 3.73 9.44 ± 1.01 7.89 ± 2.57 9.78 ± 0.44

*; Indicate the significance of each group by time period (p<0.05)

Claims (11)

A food composition comprising guarana extract, vitamins and natural extracts as active ingredients.
The food composition according to claim 1, wherein the guarana extract, vitamins and natural extract are mixed in a weight ratio of 1:0.1-0.9:0.1-0.9.
The method according to claim 1, wherein the guarana extract and the natural extract are extracted with any one extraction solvent selected from the group consisting of water, an organic solvent, and a mixed solvent thereof, and the organic solvent is alcohol, petroleum ether, hexane, benzene. , chloroform, methylene chloride, ether, ethyl acetate, and will be selected from the group consisting of acetone, the food composition.
The food composition according to claim 1, wherein the alcohol is selected from the group consisting of methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, n-butanol, and mixed solvents thereof.
The food composition of claim 4, wherein the ethanol is an ethanol solution of 50 to 99% by volume.
The food composition according to claim 1, wherein the vitamin is selected from the group consisting of vitamin B group, vitamin C, vitamin E, and combinations thereof.
The food composition according to claim 1, wherein the natural product is selected from the group consisting of tangerines, sweet persimmons, strawberries, melons, apples, omija, tomatoes, and combinations thereof.
The food composition according to claim 1, wherein the food composition is a composition for recovery from fatigue.
The food composition according to claim 1, wherein the food composition is an antioxidant composition.
The food composition according to claim 1, wherein the food composition is a composition for protecting DNA damage.
The food composition according to claim 1, wherein the form of the composition is selected from the group consisting of powders, granules, tablets, capsules and beverages.

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