KR100510987B1 - Beverage Composition - Google Patents

Abstract

The present invention comprises, as a main component, ⒜ citric acid and / or ⒝ zinc and L-arginine; Sweeteners; Acidulants; Mating agent; Preservatives; It relates to a beverage composition, characterized in that it contains at least two of the group consisting of a flavoring agent and a natural fruit juice.

Description

Beverage Composition

The present invention relates to a beverage composition and a preparation method thereof. More specifically, the main components of the present invention include Citric acid and / or Zinc and L-Arginine, and as an auxiliary component, a reinforcing agent; Sweeteners; Acidulants; Mating agent; Preservatives; It relates to a beverage composition characterized in that it contains two or more of the group consisting of a flavoring agent and natural fruit juice and a method for producing the same.

Citric acid is a free acid in lemons, plums and citrus fruits. It is a basic organic acid extracted from fruits and used as a soft drink or as a food additive. Recently, it is produced by fermentation using microorganisms using sugar (eg, starch or starch meal) as a substrate. Generally, citric acid is a crystalline white powder, soluble in water, and when added to soft drinks, it gives a fresh taste. Citric acid is also an important component of metabolism in higher animals as a component of the tricarboxylic acid (TCA) cycle in vivo. In addition, it is known to exhibit the effect of promoting calcium absorption in the body and bactericidal effect. It is mainly used as a food additive that has a sour taste in fruit juices, soft drinks, and diuretic beverages, and is widely used as medicines, analytical reagents, and cosmetic raw materials.

Zinc is present in all somatic cells in vivo and is one of more than 200 enzymes. In fact, zinc functions more than any other mineral in enzyme reactions. It is also necessary for the proper activity of many human hormones, including thyroid hormones, insulin growth hormones and sex hormones. In general, the adult human body contains a total of 1.4 to 2.5g of zinc, which is mainly stored in muscle, and is highly concentrated in red blood cells and white blood cells. Tissues with particularly high zinc levels include bone, skin, kidneys, liver, pancreas, retina and prostate. The standard daily intake of zinc is 1 to 15 mg, which is a well known source of oysters, but is also present in relatively high concentrations in other crustaceans, fish and red meat. Many plant foods, such as grains, beans, nuts, and seeds, also contain zinc in sufficient concentrations. However, zinc contained in these foods is poorly used because it combines with phytin (fibrous compound) acid to form an insoluble zinc phytate salt complex that is not absorbed. There are many zinc forms to choose from. Most clinical studies have used zinc sulphate, but the body absorbs and uses many other forms better. For example, picolinate and acetate, citrate, glycerin, or a combination of monomethionine and zinc are examples.

L-arginine is known to play an important role in cell regeneration, immune function, hormone normalization and protein metabolism. In particular, L-arginine promotes the anticancer activity of cytotoxic T cells and natural killer cells, thereby delaying the growth and metastasis of tumors and cancer cells. Has been reported (Novaes et al. Arch. Latinoam. Nutr., 49 (4), pp301-8, 1999). In addition, L-arginine is effective in the liver diseases such as cirrhosis and fatty liver through the detoxification role of the liver, and is also a major component to increase the activation of sperm production. Therefore, L-arginine is used as a useful ingredient in many medicines, dietary supplements.

As listed above, citric acid, zinc and L-arginine, which are used as the main components of the present invention, are commercially available in the form of powders, chelating compounds, capsules, etc. alone or in the form of mixtures with other ingredients. There is a risk that the activity of the active ingredient in the loss considerably, there is a problem in terms of absorption efficiency after taking.

Therefore, the present inventors induce the killing of cancer cells by treating the known effects of citric acid, zinc and L-arginine on the living body, and in particular, treating cancer cells with a composition containing citric acid and / or zinc and L-arginine as main ingredients. (See Korean Patent Application No. 2002-67291 and Korean Patent Application No. 2002-67293) to provide a beverage composition containing citric acid and / or zinc and L-arginine as main ingredients, which have beneficial effects on the human body. It is easily absorbed and can be taken easily.

The inventors have discovered that compositions comprising citric acid and / or zinc and L-arginine have beneficial effects on humans, in particular anticancer effects.

Accordingly, the present invention comprises, as a main component, ⒜ citric acid and / or ⒝ zinc and L-arginine; Sweeteners; Acidulants; Mating agent; Preservatives; It provides a beverage composition characterized in that it contains two or more kinds from the group consisting of flavoring agent and natural juice.

In addition, the present invention comprises a) admixing 1 to 20% by weight citric acid and / or 0.001 to 5% by weight zinc with 0.003 to 60% by weight L-arginine, followed by mixing 1 to 30% by weight Preparing a syrup by adding and dissolving a sweetener and adding 0.05 to 1% by weight of an acidulant;

b) adding 0.001 to 0.3% by weight of a reinforcing agent in purified water to dissolve it;

c) adding and stirring the liquid of b) to the liquid of a);

d) dissolving 0.01 to 3% by weight of the mating agent in passionate water;

e) dissolving 0.001 to 0.1% by weight of a preservative while stirring in passion water;

f) mixing the solution of d) and e) into the solution of c) and completely dissolving the solution, and then adding flavoring agent and natural fruit juice, homogenizing it with gentle stirring, and then adding a suitable amount of purified water to obtain a beverage composition. It provides a method for producing a beverage composition.

The present invention is composed of ⒜ citric acid and / or ⒝ zinc and L-arginine as main ingredients, and as an adjuvant, various reinforcing agents, sweetening agents, acidifying agents, preservatives, flavoring agents, natural juices, etc. are added to facilitate absorption in vivo. And it provides a beverage composition in the form of a liquid, characterized in that exhibits high physiological activity.

Citric acid as a main component of the present invention is a component of the TCA circuit in vivo to increase the metabolism of higher animals to exhibit a fatigue recovery effect, and to promote calcium absorption and sterilization in vivo. As another main component of the present invention, zinc has a wide range of effects. This is because zinc is involved in many enzymes and the functioning of the human body. Among them, the typical effect of zinc is to increase the activity of immune function, wound healing, sensory function and sexual function.

L-arginine, which is contained as citric acid and / or zinc as a main component of the present invention, plays an important role in vivo as an amino acid that enhances cell regeneration, immune function, hormone normalization, protein metabolism and anticancer activity.

As indicated above, the present inventors describe epithelial ovarian cancer in order to prove that a composition prepared with citric acid and / or zinc and L-arginine exhibiting beneficial effects on the human body, in particular, has anticancer effects. I would like to. However, one of ordinary skill in the art would recognize that the effects of citric acid and / or compositions prepared with zinc and L-arginine on epithelial ovarian cancers described herein by way of example may be equally applicable to other types of cancer. will be.

Epithelial ovarian cancer is the most common type of ovarian cancer as well as the leading cause of malignancy in gynecology. The highest mortality rates in patients with epithelial ovarian cancer are due to the fact that most patients are diagnosed at an advanced stage of cancer due to the latent progression of epithelial ovarian cancer (Ozols, RF, Semin. Oncol., Vol. 22, pp 61-66, (1995)). Although ovarian cancer is treated by surgical removal of the tumor or by aggressive chemotherapy, survival of ovarian cancer patients is only 20-30% due to drug resistance.

To date, little is known about the risk factors and causes of epithelial ovarian cancer. However, many studies have shown that there is a common relationship between the frequency of ovulation and the development of epithelial ovarian cancer, which suggests that young and older women (early menopause and late menopause), Increase in unmarried and unborn women (Franceshi et al., Int. J. Cancer, vol. 49, pp57-60, (1991); Taylor et al., Cancer, vol. 12, p1207-, (1959); Fraumeni et al., J. Natl. Cancer Inst., Vol. 42, p455-, (1969); Dorn et al., Public Health Monogr. 56, PHS Publication No. 590, (1959); Weiss et al., J. Natl. Cancer Inst., vol. 58, p913-, (1977); (Nergri et al., Int. J. Cancer, vol. 49, pp 50-56, (1991)). Oral contraceptives are generally known to have a protective effect against the development of epithelial ovarian cancer (Stanford, JL, Contraception 43, pp543-556, (1991); Negri et al., Int. J. Cancer, vol. 49, pp 50-56, (1991); Franceshi et al., Int. J. Cancer, vol. 49, pp 61-65, (1991)) A potent theory explaining the development of epithelial ovarian cancer is that the ovarian epithelial surface divides and recovers repeatedly during ovulation (Fathalla, MF). , Lancet., Vol. 2, p163-, (1971)) In the process of restoring the epithelial surface, the modified epithelial cells spontaneously mutate, the tumor suppressor genes are inactivated, and the oncogenes are caused by carcinogens. It is easy to be activated.

On the other hand, as a known semen function and anticancer effect, human semen is known to protect the seminal plasma components of sperm after sexual intercourse due to cytotoxicity of lymphocytes (Stities et al., Nature, vol. 253, pp727-729, (1975); James et al., Immunol., Vol. 6, pp61-70, (1985)), have been reported to inhibit the development of humoral immunity and tumor growth in vivo. (Anderson et al., Immunol., Vol. 128, pp535-539, (1985); Michaelis et al., Anticancer Drugs, vol. 11, pp369-376, (2000)), and also in cervical epithelial carcinoma cells Since it affects the production of metalloproteinases (MMP) -2 and MMP-9 mRNA, it is known that semen during sexual activity can affect cervical cancer progression (Jeremias et al., Am J. Obstet.Gynecol., Vol. 181, pp 591-595, (1999). Recently, ribonuclease (BS-RNase) among bovine semen has been shown to be capable of inducing apoptosis depending on the time and concentration of exposure to human lymphocytes and human tumor cells. BN-RNase is a substance that shows high efficacy against neuroblastoma (NB) cells resistant to chemotherapeutic drugs (Cinatl et al., Anticancer Res., Vol. 20, pp853-859, (2000); Cinatl et. al., Int. J. Oncol., vol 15, pp 1001-1009, (1999)).

In addition, the ecological study of anti-cancer effects of semen, Gjorgov conducted a variety of case-control studies to investigate the correlation between reduced exposure of human semen and the development of breast cancer. For example, we compared the risk of breast cancer in women who used a block contraceptive method (condom) versus women who used a non-blocking method (contraceptives, intra uterine device (IUD), rhythm or fallopian ligation). In contrast, women using blocking contraceptive methods (condoms) were 5.2 times more likely to develop breast cancer (Gjorgov et al., Folia Med., Vol. 40, pp 17-23, (1998)).

The main components of semen exhibiting the anticancer effects shown above include albumin, lactoferrin, transferring factors, immunoglobulins, acid phosphatase, L-carnitine, and L-arginine. , L-Histidine, citric acid, fructose, fructose, magnesium, zinc, prostaglandin and glycerophosphocholine.

As mentioned above, from the prior knowledge of the anticancer effects of epithelial ovarian cancer and semen, the present inventors have found that in epithelial ovarian cancer, reduced exposure to human semen during ovulation is a pathological risk factor for the progression of epithelial ovarian cancer. I decided it would be one of Thus, human semen components may contribute to the removal of malignant transformed epithelial cells.

In this regard, the present inventors have studied human semen components affecting cancer cells in vitro, and citric acid among human semen components acts on the mitochondria of cancer cells and breaks down the mitochondria. It has been found to protect more and induce the death of cancer cells, and to increase the anticancer activity of cytotoxic T cells and natural killer cells in citric acid and / or zinc and conventional cancer treatments to delay the growth and metastasis of tumors and cancer cells. It has been found that mixing the known L- arginine together increases the anticancer effect (see Korean Patent Application No. 2002-67291 and Korean Patent Application No. 2002-67293). Therefore, the present inventors prepared a composition containing citric acid and L-arginine and treated with SKOV-3, a human ovarian adenocarcinoma, and U87, a human brain glioblastoma, and confirmed that the effect of inducing apoptosis and killing cancer cells is high. As a result of preparing a composition containing zinc and L-arginine and treating it with SKOV-3 and U87, it was confirmed that the effect of inducing apoptosis and killing cancer cells was high (see the following figure). That is, when citric acid and L-arginine or a composition containing zinc and L-arginine were treated to cancer cell lines SKOV-3 and U87, citric acid and / or zinc and L-arginine, which are the main components of the present invention, were treated. It can be seen that the containing composition exhibits anticancer action inducing apoptosis of cancer cells.

As mentioned above, the known effects of citric acid, zinc and L-arginine, and in particular the anticancer effect of citric acid and / or compositions containing zinc and L-arginine, indicate that the beverage compositions of the present invention exhibit beneficial effects on the human body. To show.

Therefore, the present invention contains two or more of the reinforcing agent, sweetening agent, acidulant, copulating agent, preservative, flavoring agent, natural fruit juice as auxiliary components in purified water homogeneously dissolved in citric acid and / or zinc and L- arginine as the main component It provides a liquid beverage composition and a method for producing the same.

In one embodiment, the method for producing a beverage composition of the present invention is as follows.

⒜ 1-20 wt% citric acid and / or 0.001-5 wt% zinc and 0.003-60 wt% L-arginine are mixed with stirring, followed by dissolution of 1-30 wt% sweetener After the addition, 0.05 to 1% by weight of acidifier is added to prepare syrup.

⒝ Separately, 0.001 to 0.3% by weight of a reinforcing agent is added to the purified water to dissolve it.

액 A solution of ⒝ is added to the syrup of ⒜ prepared above, followed by stirring, and then, as an additional additive, 0.01 to 1% by weight of caffeine anhydrous or 0.1 to 1% by weight of flavoring agent may be added thereto.

⒟ Separately, 0.01 to 3% by weight of a mating agent is added to the passion solution to dissolve it.

0.00 Also, 0.001 to 0.1% by weight of a preservative is dissolved separately in passion water.

에 Mix the solution of ⒟ and 에 to the solution of ⒞, prepared above, and dissolve completely, add a flavoring agent or natural fruit juice, stir it slowly, pass it through a homogenizer (NISSEI), sufficiently homogenize, and then 100 mesh After filtering by (mesh) and adding a suitable amount of purified water, sterilization with an instant sterilizer (Miura, Japan) to obtain a 100ml beverage composition to prepare a beverage composition of the present invention.

Zinc used as a main component in the present invention is not limited to these, but may be used zinc chloride, zinc sulfate, zinc acetate, and the like, preferably zinc chloride.

Also. Auxiliary components in the present invention are not limited to these, but may be used as a reinforcing agent, sweetener, acidulant, copulation agent, preservative, flavoring agent and natural juices according to each formulation.

As the reinforcing agent used in the present invention, as vitamins, vitamins A, B ₁ , B ₂ , C, D ₂ , B ₃ , E, and folic acid can be used for all vitamins contained in food or food additives. For example, use a mixture of vitamin B ₂ or vitamin C. L-lysine, DL-methionine, L-valine, biotin, L-cysteine, L-cystine, L-tryptophan, L-threonine, L-phenylalanine, L-glycine, etc. can be used as amino acids. Calcium salts include calcium pantothenate, calcium gluconate, calcium carbonate, calcium citrate, vegetable calcium, animal calcium, oyster shell powder calcium and the like.

The sweetening agent used in the present invention may be used all sweeteners contained in food or food additives, such as high fructose, white sugar, glucose, aspartame, stevioside, D-sorbitol, glycylic acid salts, preferably glucose or white sugar 2 or more types are mixed and used.

The acidulants used in the present invention may be used in all food additives such as DL- peracid, succinic acid, D-tin acid, adipic acid and the like in food or food additives, preferably DL-peracid or D-tin acid and the like use.

The mating agent used in the present invention may use all the mating agents carried in the food or food additives, preferably, a mixture of two or more kinds of sodium chloride, disodium succinate or sodium L-glutamate, the amount is It can be added or subtracted according to the amount, type of drink and the amount of each active ingredient.

Preservatives used in the present invention include benzoic acid and its derivatives (e.g. sodium benzoate), dehydroacetic acid and its salts (e.g. sodium dehydroacetic acid), sorbic acid and its salts (e.g. calcium sorbate), etc. All the preservatives entrained in the additive can be used alone or in combination of two or more thereof.

Flavoring agents used in the present invention may be used alone or in combination of two or more of the apple flavor, strawberry flavor, orange flavor, mixed fruit flavor, yogurt flavor, drink flavor as a natural or artificial flavor.

As the natural juices used in the present invention, natural juice concentrates such as apples, pears, pineapples, tangerines, grapefruits, peaches, apricots, strawberries, lemons, plums and melons may be used alone or in combination of two or more thereof.

The beverage composition of the present invention prepared according to the above production method illustratively consists of each component as follows. It contains 1 to 20 weight percent citric acid and / or 0.001 to 5 weight percent zinc and 0.003 to 60 weight percent L-arginine as main components. In addition, as an auxiliary component, vitamins as reinforcing agents contain 0.001 to 0.1% by weight of vitamin B ₂ or 0.001 to 0.1% by weight of vitamin C, and calcium salts as reinforcing agents are 0.001 to 0.1% by weight of calcium gluconate or 0.001 to 0.1. Contains calcium pantothenate by weight. Sweeteners contain 1.0 to 20% by weight of white sugar and 1.0 to 10.0% by weight of glucose, and acidifiers contain 0.05 to 1% by weight of DL-peracid. The mating agent contains 0.01-1.0 wt% sodium L-glutamate or 0.01-1.5 wt% sodium chloride or 0.01-1.0 wt% disodium succinate. Preservatives contain 0.05 to 0.1 wt% sodium benzoate and 0.05 to 0.1 wt% sorbic acid, and 0.01 to 1.0 wt% anhydrous caffeine or 0.1 to 1 wt% flavoring agent or natural juice can be added if necessary. have.

The invention will be more specifically illustrated by the following experimental examples and examples. However, these experimental examples and examples are merely embodiments of the present invention and do not limit the scope of the present invention.

<Example 1>

Beverage composition of the present invention

⑴

In 100 ml (100 g)

10 g citric acid

30 g of L-arginine

10g per bag

2.5 g of glucose

DL-apple acid 0.3 g

0.005 g of vitamin B ₂

0.005 g of vitamin C

Calcium Gluconate 0.015g

0.055 g of sodium L-glutamate

Sodium chloride 0.035g

Sodium benzoate 0.06 g

Solvate 0.01g

100% of purified water

Method for preparing a beverage composition of the present invention 1) mixed with 10 kg citric acid (Sigma. USA), L- arginine 30 kg (Ajinomoto, Japan) while stirring in 30 liters of passion distillate, 10 kg per bag, 2.5 kg of glucose was dissolved After the addition, 0.3 kg of DL-peric acid was added to prepare a syrup for 15 minutes while maintaining at 90 ° C.

2) 1) and are separate from, and in purified water 5ℓ added vitamin B ₂ 5g, 5g of vitamin C and calcium gluconate 15g sequentially and dissolved while maintaining 50 to 60 ℃.

3) The liquid of 2) was added to the liquid of 1), and it stirred for 1 hour, maintaining 50-60 degreeC continuously.

4) 55 g of sodium L-glutamate and 35 g of sodium chloride were dissolved in 10 L of a passion solution.

5) 10 g of sorbic acid and 60 g of sodium benzoate were added and dissolved in 5 liters of previously prepared passion solution (preservative).

6) Mix the solution of 4) and 5) in the solution of 3) and completely dissolve it. After homogenizing with gentle stirring, filter it with 100 mesh and add the purified water quantity to adjust the total amount to 100ℓ. Instant sterilization at 95 ℃ for 30 seconds through a sterilizer and then filled into a 100ml bottle using an automatic charger.

⑵

In 100 ml (100 g)

1 g of zinc chloride

3 g of L-arginine

15g per bag

Glucose 3.0g

0.3 g D-Tartrate

0.01 g of vitamin B ₂

0.01 g of vitamin C

Calcium Pantothenate 0.02g

0.03 g of caffeine anhydrous

0.055 g of sodium L-glutamate

Disodium Succinate 0.04g

0.055 g sodium benzoate

Solvate 0.01g

100% of purified water

1) 1 kg of zinc chloride (Sigma. USA) and 3 kg of L-arginine (Ajinomoto, Japan) were mixed with 30 liters of passion solution, and 15 kg of white sugar and 3 kg of glucose were dissolved and 0.3 kg of D-tin acid. The syrup was prepared for 15 minutes, adding and maintaining 90 degreeC.

2) Apart from 1), 10 g of vitamin B _2, 10 g of vitamin C and 20 g of calcium pantothenate were sequentially added to 5 L of purified water, and dissolved at 50 to 60 ° C.

3) The liquid of 2) was added to the liquid of 1), and it stirred for 1 hour, maintaining 50-60 degreeC continuously.

4) 55 g of sodium L-glutamate and 40 g of disodium succinate were dissolved in 10 liters of diluent.

5) 30 g of anhydrous caffeine was dissolved in 5 liters of passion solution.

6) 55 g of sodium benzoate and 10 g of sorbic acid were added and dissolved in 5 liters of previously prepared zeolite, which was dissolved (preservative).

7) Mix the solution of 4), 5) and 6) in the solution of 3) and completely dissolve it. After homogenizing with gentle stirring, filter it with 100 mesh and add the purified water quantity to 100ℓ. After sterilization for 30 seconds at 95 ℃ through an instant sterilizer and then filled into a 100ml bottle using an automatic charger.

⑶

In 100 ml (100 g)

15 g citric acid

Zinc Chloride 3g

45 g of L-arginine

10g per bag

2.5 g of glucose

DL-apple acid 0.3 g

0.005 g of vitamin B ₂

0.005 g of vitamin C

Calcium Gluconate 0.015g

0.055 g of sodium L-glutamate

Sodium chloride 0.035g

Sodium benzoate 0.06 g

Solvate 0.01g

100% of purified water

1) Mix 15 kg of citric acid (Sigma. USA), 3 kg of zinc chloride (Sigma, USA) and 45 kg of L-arginine (Ajinomoto, Japan) while stirring in 30 l of passion distillate, and then 10 kg per bag and 2.5 kg of glucose. After the solution was added and dissolved, 0.3 kg of DL-peric acid was added thereto to prepare a syrup for 15 minutes while maintaining 90 ° C.

2) 1) and are separate from, and in purified water 5ℓ added vitamin B ₂ 5g, 5g of vitamin C and calcium gluconate 15g sequentially and dissolved while maintaining 50 to 60 ℃.

3) The liquid of 2) was added to the liquid of 1), and it stirred for 1 hour, maintaining 50-60 degreeC continuously.

4) 55 g of sodium L-glutamate and 35 g of sodium chloride were dissolved in 10 L of a passion solution.

5) 10 g of sorbic acid and 60 g of sodium benzoate were added and dissolved in 5 liters of previously prepared passion solution (preservative).

6) Mix the solution of 4) and 5) in the solution of 3) and completely dissolve it. After homogenizing with gentle stirring, filter it with 100 mesh and add the purified water quantity to adjust the total amount to 100ℓ. Instant sterilization at 95 ℃ for 30 seconds through a sterilizer and then filled into a 100ml bottle using an automatic charger.

<Example 2>

Sensory test of the beverage composition of the present invention

Table 1 shows the results of three repeated sensory tests on the degree of preference for 30 sensory test agents for the beverage compositions of Example 1, ⑵ and ⑶ prepared as described above.

Sensory test of the beverage composition of the present invention Inspection items 의 of Example 1 의 of Example 1 의 of Example 1 color 4.0 4.1 4.2 flavor 4.1 4.0 3.9 incense 3.8 3.9 4.0 Mouth feel 4.0 4.1 4.1 Overall preference 4.2 4.0 4.1

^* 30 people for sensory evaluation

^** Inspection method: 5-point scale method

^*** 1: I hate it. 2: No. 3: Normal 4: good. 5: very good.

Experimental results of Table 1 shows that the beverage composition of the present invention can be usefully used as a beverage that can be preferred in terms of sensuality.

Experimental Example

세포 Cell viability measurements following treatment with compositions containing citric acid and L-arginine.

In order to investigate the effects of the composition containing citric acid and L-arginine on the growth and survival of cancer cells as a main component of the present invention, the cell viability was measured by treating the cancer cell line with a composition prepared by varying the content of the main component. Cell lines include SKOV-3 (human ovarian adenocarcinoma, ATCC number: HTB-77), OVCAR-3 (human ovarian adenocarcinoma, ATCC number: HTB-161), and U87 (human brain glioblastoma, ATCC number: HTB-14 ) Was used.

The cells were placed in 96-well plates to 3 × 10 ³ cells / well and incubated for 12 hours. As culture medium, DMEM (Dulbecco's modified) was added 10% (v / v) of heat-inactivated FBS (fetal bovine serum), 100 μg / ml of streptomycin, 100 U / ml of penicillin and 100 μg / ml of L-glutamine. Eagle's medium, Life Technology, Inc., USA). After treating and classifying the composition prepared by varying the content of citric acid and L-arginine in each cultured cell line, the cells were incubated for 24 hours in a state where 5% carbon dioxide and 95% oxygen were supplied at 37 ° C, and then cell viability. Was measured.

Cell viability was measured by known MTT (3- (4,5-dimethylthiazole-2-yl) 2,5-diphenyl-2H-tetrazolium bromidine) assay (Hansen, MB et al., J Immunol.Methods, 172, 203-210 (1989). 20 μl MTT solution [MTT in 10 mg / ml in phosphate buffered saline (PBS)] was added to each well and the plate was incubated at 37 ° C. for 4 hours. After removing the medium, formazan crystals were dissolved in 200 µl of DMSO (dimethyl sufoxide, dimethyl sulfoxide) and added to each well. This was shaken for 10 minutes at room temperature and mixed and measured at 540 nm using a Bio-Rad Model 3550 Microplate Reader (Microplate Reader, Richmond, CA.). At this time, wells without DMEM-FBS and MTT added and the composition was used as a control. Experimental results are shown in FIGS. 1 and 2.

As can be seen from Figures 1 and 2, the viability of cell lines SKOV-3 and U87 treated with a composition containing citric acid and L-arginine as a main component decreased with increasing the content of citric acid and L-arginine. In particular, when the content of citric acid and L-arginine is 0.3% by weight or more, the survival rate is reduced to 50% or less, and when the content of citric acid and L-arginine is 0.8% by weight or more, the survival rate is reduced to 20% or less.

측정 Measurement of cell viability following treatment with composition containing zinc and L-arginine

In order to investigate the effect of the composition containing zinc and L-arginine on the growth and survival of cancer cells as another main component of the present invention, the cell viability was measured by treating the cancer cell line with a composition prepared by changing the content of the main component. SKOV-3 and U87 were used as cell lines.

The cells were placed in 96-well plates to 3 × 10 ³ cells / well and incubated for 12 hours. As culture medium, DMEM (Dulbecco's modified) was added 10% (v / v) of heat-inactivated FBS (fetal bovine serum), 100 μg / ml of streptomycin, 100 U / ml of penicillin and 100 μg / ml of L-glutamine. Eagle's medium, Life Technology, Inc., USA). After treating and sorting the composition prepared by varying the content of zinc and L-arginine in each cell line, the cells were incubated for 24 hours at 37 ° C. with 5% carbon dioxide and 95% oxygen, and then cell viability. Was measured.

Cell viability was measured by known MTT (3- (4,5-dimethylthiazole-2-yl) 2,5-diphenyl-2H-tetrazolium bromidine) assay (Hansen, MB et al., J Immunol.Methods, 172, 203-210 (1989). 20 μl MTT solution [MTT at 10 mg / ml in (phosphate buffered saline, phosphate buffered saline)] was added to each well and the plate was incubated at 37 ° C. for 4 hours. After removing the medium, formazan crystals were dissolved in 200 µl of DMSO (dimethyl sufoxide, dimethyl sulfoxide) and added to each well. This was shaken for 10 minutes at room temperature and mixed and measured at 540 nm using a Bio-Rad Model 3550 Microplate Reader (Microplate Reader, Richmond, CA.). At this time, wells without DMEM-FBS and MTT added and the composition was used as a control. Experimental results are shown in FIGS. 3 and 4.

As can be seen from Figures 3 and 4, the viability of cell lines SKOV-3 and U87 treated with a composition containing zinc and L-arginine decreased with increasing content of zinc and L-arginine as active ingredients. It was. In FIG. 3, the survival rate of the composition containing zinc chloride and L-arginine is reduced to 50% or less when the content of the component is about 0.008% by weight or more, and the survival rate is 20 when the content of the component is about 0.01% by weight or more. Decreased below%. In addition, in the composition containing zinc sulfate and L-arginine, the survival rate was reduced to 50% or less when the content of the component was about 0.004% or more, and the survival rate was 20% or less when the content of the component was about 0.01% by weight or more. Reduced to. In FIG. 4, the survival rate of the composition containing zinc chloride and L-arginine is reduced to 50% or less when the content of the component is about 0.005% by weight or more, and the survival rate is 20 when the content of the component is about 0.008% by weight or more. Decreased below%.

The present invention is a beverage composition comprising citric acid and / or zinc and L-arginine as a main ingredient, and as an auxiliary ingredient, a reinforcing agent, a sweetener, an acidulant, a coagulant, a preservative, a flavoring agent and a natural fruit juice, and the like. As a method, it is easily absorbed into the body and can be conveniently used to give a beneficial effect to the human body.

Figure 1 shows the effect of the composition containing citric acid and L-arginine on the human ovarian adenocarcinoma SKOV-3 as the main component of the present invention as a cell viability through the MTT assay.

Figure 2 shows the effect of the composition containing citric acid and L- arginine on the human brain glioblastoma U87 as the main component of the present invention as a cell viability through the MTT assay.

Figure 3 shows the effect of the composition containing zinc and L-arginine as the main component of the present invention on human ovarian adenocarcinoma SKOV-3 as cell viability through the MTT assay.

Figure 4 shows the effect of the composition containing zinc and L-arginine as a main component of the present invention on human brain glioblastoma U87 as a cell viability through the MTT assay.

Claims (11)

1 to 20% by weight citric acid, 0.001 to 5% by weight zinc and 0.003 to 60% by weight L-arginine as a main component; Sweeteners; Acidulants; Mating agent; Preservatives; Beverage composition characterized in that it contains 100% composition amount of two or more kinds from the group consisting of flavoring agent and natural fruit juice. delete delete The method of claim 1, wherein the reinforcing agent is an auxiliary component; A beverage composition comprising two or more selected from the group consisting of amino acids and calcium salts. The sweetener of claim 1, wherein the sweetening agent is an auxiliary component; Sucrose; glucose; Asparam; Steviosides; A beverage composition comprising two or more selected from the group consisting of D-sorbitol and glycidyl acid salts, and mixing them. The acidic agent of claim 1, wherein the acidic agent is an auxiliary component; Succinic acid; Beverage composition characterized in that it contains at least one selected from the group consisting of D-tin acid and adipic acid. The method of claim 1 wherein the mating agent is an auxiliary component sodium chloride; A beverage composition, characterized in that it contains a mixture of two or more selected from the group consisting of disodium succinate and sodium L- glutamate. The preservative of claim 1, wherein the preservative is an auxiliary component; Sorbic acid; A beverage composition characterized by containing one or two or more selected from the group consisting of calcium sorbate and sodium dehydrotroacetate, and mixing them. The flavoring agent of claim 1, wherein the flavoring ingredient is an auxiliary ingredient; Strawberry flavor; Orange flavor; Mixed fruit flavor; A beverage composition characterized by containing only one or two or more selected from the group consisting of yogurt flavor and drink flavor. The method of claim 1, wherein the natural fruit juice as an auxiliary component is apple; ship; pineapple; tangerine; Gray fruits; peach; A beverage composition characterized by containing one or two or more selected from the group consisting of apricot and lemon. a) 1 to 20% by weight citric acid, 0.001 to 5% by weight zinc and 0.003 to 60% by weight L-arginine are mixed with agitation, followed by dissolution of 1 to 30% by weight sweetener Preparing a syrup by adding 0.05 to 1% by weight of an acidulant; b) adding 0.001 to 0.3% by weight of a reinforcing agent in purified water to dissolve it; c) adding and stirring the liquid of b) to the liquid of a); d) dissolving 0.01 to 3% by weight of the mating agent in passionate water; e) dissolving 0.001 to 0.1% by weight of a preservative while stirring in passion water; f) mixing the solution of d) and e) into the solution of c) and completely dissolving the solution, and then adding flavoring agent and natural fruit juice, homogenizing it with gentle stirring, and then adding a suitable amount of purified water to obtain a beverage composition. Process for preparing a beverage composition.