KR100440493B1 - Healthy drink comprising Alismatis Rhizoma extract and process for prepatration therof - Google Patents

Abstract

The present invention relates to a taxa health beverage and a method for manufacturing the same, and adds apple concentrate, acidulant, sweetener and flavor to the extracts of herbal extracts such as taxa extract, jujube, wolfberry and Schisandra chinensis with excellent antimicrobial, antioxidant and immune activity. There is an excellent effect of providing enhanced tax health drinks.

Description

Health drink comprising Alismatis Rhizoma extract and process for prepatration therof}

The present invention relates to a taxi health drink and a manufacturing method thereof. More specifically, the present invention relates to a tactile health beverage with improved functionality by manufacturing a mixture of herbal medicine extracts and subsidiary ingredients in a medicinal extract extract having excellent antimicrobial activity, antioxidant effect and immune activity, and a method for producing the same.

Alismatis Rhizoma , an Alismataceae plant, is a remnant group differentiated during the evolution of monocotyledonous plants, with about 13 genera and 90 species distributed worldwide. Fujian, Jiangxi, Sichuan, Guitian and It is also produced in Yunnan province, and in Korea, it is produced in the Jeonnam Suncheon region, which accounts for more than 50% of its production, in Muan, Gurye, and Sangju, Gyeongsangbuk-do, and the taxa used as a herbal medicine is Alisma plantago- aquatica L. var. orientale Samuels) is mainly grown and produced. The constituents of taxa are known as 25% starch, 7% protein, furfural, resin, essential salts, alisol A, B, C, and acetates and acetates of triterpenoids. Such taxa is known as a medicine that removes moisture, helps the intestine, and strengthens energy. It is contained a lot of potassium (K), and when the taxi is taken, potassium (K) in the urine is significantly increased. Taxa has also been reported to reduce blood cholesterol, lower blood pressure and blood sugar, and have an effective interpolation effect on hepatotoxicity induced by carbon tetrachloride. In addition, recently, the tactile powder has been used as a medicine for athlete's foot, so it is thought that various physiologically active substances are present in the tactical taxi. However, the actual research results on physiological activity of taxis are very insufficient and the development of health foods using them is insufficient.

The present inventors have completed the present invention by identifying the physiological activity such as the effects on the antibacterial, antioxidant activity and immune response control action of taxis produced in the Suncheon area by focusing on the above points and preparing the taxis as a beverage.

Accordingly, it is an object of the present invention to provide a method for producing a tax health drink. Another object of the present invention is to provide a taxi health drink.

1 is a schematic diagram showing an embodiment of a preferred manufacturing method of the present invention for each process.

Figure 2 is a photograph of a plate medium showing the growth inhibition ring produced after the addition of the tax water extract to the test strain to determine the antimicrobial activity of the tax extract of the present invention.

Figure 3a is a graph showing the growth inhibitory effect of the Bacillus subtilis strain of the present invention tack extract.

Figure 3b is a graph showing the growth inhibitory effect of Pseudomonas aerignosa strain of the present taxa extract.

Figure 3c is a graph showing the growth inhibitory effect of E. coli strains of the present invention tack extract.

Figure 3d is Salmonella typhymuriu of the present invention taxa extract ( Salmonella typhymuriu-

m ) It is a graph showing the growth inhibitory effect of the strain.

Figure 3e is a graph showing the growth inhibitory effect of the Vibrio parahaemo liticus strain of the present taxa extract.

Figure 4 is Salmonella typhymuriu according to the process of extracting the taxa ( Salmonella typhymuriu

m ) electron micrograph.

Figure 5 is a graph showing the change in peroxide value according to the addition of the tack extract (100 μl) during linoleic acid automatic oxidation for 7 days at 50 ℃.

Figure 6 is a graph showing the change in peroxide value according to the concentration of the tax extract during the automatic oxidation of linoleic acid for 7 days at 50 ℃.

Figure 7 is a graph showing the change in TBA value when the present invention is treated with a rat extract.

Figure 8a is a graph showing the effect on the proliferation of splenocytes when the present invention is treated to splenocytes.

Figure 8b is a graph showing the effect on the proliferation of B cells when the present invention is treated to B cell tack extract.

Figure 9a is a graph showing the effect of the present invention tack extract on T cell proliferation in response to homologous antigen.

Figure 9b is a graph showing the degree of inhibition of T cells in response to the tax extract of the present invention homologous antigen.

Figure 10 is a graph showing the effect of the present invention taxa extract on the cytotoxicity of T cells in response to homologous antigen.

Figure 11 is a photographic view of the tactile health beverage product according to the present invention.

The object of the present invention is to prepare a tack extract to check the antibacterial activity against food poisoning and pathogenic microorganisms, the antioxidant effect and immune activity against linoleic acid and hepatic lipids, and then the tack extract enhanced functionality using the tack extract, herbal extracts and other side ingredients Achieved by preparing a health beverage.

Hereinafter, the configuration of the present invention will be described.

The present invention comprises the steps of preparing a taxi extract using methanol, ethanol and water to investigate the antibacterial, antioxidant and immune activity of the taxi; Measuring antimicrobial activity against food poisoning and pathogenic microorganisms to investigate the antimicrobial activity of the taxa extract, and examining the morphological changes of the microorganisms according to the treatment of the taxa extract; Irradiating with respect to the in vitro antioxidant activity and lipid peroxidation inhibiting liver functions for the linoleic acid in order to examine the antioxidant activity of the extract Alismataceae; Effects of Taxa Extract on Immune Cell Proliferation, Effects of Macrophage Cells on Nitric Oxide Production, Proliferative Response to Homologous Antigens, Inhibition of T Cell Cytokine Secretion by Homologous Antigens, and T Cell Killer Action on Homologous Antigens Investigating; Preparing water and medicinal extracts by extracting and concentrating water for 10 times with water twice as much as 10 times in taxa and herbal medicines at 80 ° C. for 5 hours; Weighing and mixing the tactile extract, the herbal extract, and the subsidiary apple juice, acidulant, sweetener, and flavoring according to the selected optimal blending ratio; Flavoring and applying to the sample of the step and filtering with a 1 μm membrane filtration filter; Sterilizing the filtrate at 93 ° C. for 15 seconds and then filling the vessel; The filled sample is sterilized at 80 ° C. for 20 minutes and then commercialized. The step of applying the above means that the mixing amount of the respective materials is “fit to a predetermined capacity”, and the weighing and mixing of each material The amount of mixed sugar is precisely controlled according to a predetermined blending ratio. The mortgage is a sugar having intermediate functional properties of known starch and starch syrup. About 20, starch is partially hydrolyzed, purified and concentrated by enzymes. Mortgage is a well-known food material widely used in the food processing field because of its excellent formation and resistance to browning. It is used in coffee primer, food extender and so on. In addition, for example, strawberries contain a large amount of glucose, including low sugar or fructose, and malic acid, citric acid, tartaric acid, and the like provide a sweet and sour taste of strawberries.

Alismatis Rhizoma of the present invention purchased and used harvested in Suncheon, Jeonnam in 1998. The strains Bacillus subtilis , Pseudomonas aeruginosa , Escherichia coli , Salmonella typhimurium , Escherichia coli O-157 and Vibrio parahaemolyticus were used for antimicrobial testing. Nutrient broth (agar) was used for the culture of various microorganisms, and 3% NaCl was added for Vibrio parahaemoliticus .

Mice (BALB / c, C57BL / 6) used to investigate the immune activity of taxa were purchased from the Korea Experimental Animal Center (negative group Chungbuk), and 8-12 weeks old females were used. It was bred while supplying tea distilled water. RPMI 1640 medium, antibiotic antimicrobial was used Gibco BRL (Grand Island, NY, USA) products, FCS (fertal calf serum) was used PAA products. LPS and 2-ME (2-mercaptoehtan

ol), NaHCO 3, N-1-naphthyl-ethylen-diamine and sulfanamide were used by Sigma. In addition, the reagents used in the cytotoxicity assay (Cyto Tox 96). Non-radioactive Cytotoxicity Assay and Reagents Used to Measure Cell Proliferation (Cell titer96) Aqueous One Solution Cell Proliferation Assay was used for Promega, and the antibodies required for cytokine measurement were used for Pharmingen. Sephadex G-10 used for B cell separation was also used by Pharmingen. As a cell line for cell culture, RAW264.7, a mouse mononuclear / macrophage cell line, was purchased from Korea Cell Line Bank (Seoul National University College of Medicine). Medium for the cell culture was used by adding NaHCO ₃ 2g / L, penicillin 100unit / ml, streptomycin 100mg / ml to RPMI 1640 medium.

Alismatis Rhizoma extract used to measure antimicrobial activity and antioxidant activity was added at a rate of 1500 ml per 100 g of taxa using solvents such as methanol, ethanol and water, and methanol and ethanol at 80 ° C using a reflux condenser. , Water was prepared by extracting at 100 ℃ for 2 hours and concentrated to 0.5ml per g of taxi.

Apple concentrate (SP-6411, Bx 50) used as an ingredient in the preparation of the taxi beverage of the present invention was prepared by Myung Shin Hwaseong, and citric acid and vitamin C used as an acidulant were purchased from Hongsung Pharmaceuticals. The liquid fructose (high fructose) used as a sweetener is Doosan's product and the mortgage (70) is manufactured by Samyang Genex and the fragrance (ruitmix FLA B-8718) was purchased from Hanbul. The formulation and ratio of the extracts of the extract, the herbal extract, and the subsidiary ingredient were selected by the sensory evaluation after the preparation of the prototype, and the optimal magnification was selected. Herbal extracts such as 0.1-1.0g / 100ml and Schisandra chinensis extract 0.1-0.5g / ml, apple juice 1.0% -2.0g / 100ml, citric acid 0.01-0.1g / 100ml, vitamin C 0.01-0.05g / 100ml, liquid fructose 10-15 g / 100ml, mortgage 1.0-5.0g / 100ml, and the fragrance | flavor 0.05-0.15g / 100ml were added and prepared.

In the present invention, the health of the taxi is in harmony with the unique taste and sourness of the taxi, and the fruit flavor is in the form of a brown beverage. The present invention is characterized by having a pH of 3-4, preferably 3.58, and brix of 12-13 ° Bx, preferably 12.2 ° Bx. In addition, the present invention is a small amount of precipitate is produced even after 5 months at 4 ℃, room temperature, 37 ℃ in the tax health beverage is another quality characteristic is stable.

Preferred method of manufacturing the present taxi health beverage is shown in FIG.

Hereinafter, the specific method of the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited only to these Examples.

Experimental Example 1: Investigation of antimicrobial effect of taxa extract

In order to investigate the antimicrobial effect of taxa extracts, the effects of antimicrobial activity, microbial growth, and microbial morphology were investigated.

The measurement of antimicrobial activity modified Farag's method. That is, the growth medium containing 1.5% of agar is thinly spread on the bottom of a petri dish, and the growth medium containing 0.6% of agar is poured on it to make a double flat medium, and then each strain is placed on the flat medium. Smeared. The strain was placed on a plate medium coated with a certain amount of the tackifier extract 0.8cm in diameter filter disk (disc) and incubated for 24 hours at 36 ℃, the resulting growth inhibition was measured to investigate the antibacterial activity.

Microbial growth was measured using the liquid medium dilution method. Inoculate 5 ml of growth medium into the test tube and inoculate 1% of the cell culture cultured up to the middle of the logarithmic season, and then add tac extract extract to 30 to 200 µl per 5 ml of medium and incubate at optimum temperature of each strain. The extent of growth was measured by absorbance at 660nm using a spectrophotometer.

The morphological changes of microorganisms by Alixmatis Rhizoma extracts were treated with food extract microorganisms and their morphological changes were observed by electron microscopy. Incubate Salmonella typhimurium as a decay microorganism in a culture medium for 48 hours, add 100 microliters of taxa extract per ml of culture strain to the culture strain, leave for 3 hours, and centrifuge to isolate the cells. 0.05M phosphate buffer The cells were diluted with and fixed to 0.45 μl membrane filter. Soak overnight in 5% glutaaldehyde (glutaaldehyde) solution, wash with sterile water, and then immerse in 30 ~ 100% ethanol one by one in order to dehydrate, immerse in isoamylacetate for about 30 minutes to dry and electron microscope The photographing sample was prepared and observed.

Experimental results, as shown in Figure 2 and Table 1, the antimicrobial activity of the taxa extract, water extract showed stronger antibacterial activity than methanol and ethanol extract. Also had antimicrobial activity appeared strongly to all tested strains except for Escherichia coli O-157 strain clear zone (clear zone) of Alismataceae extract of each test strain Vibrio parahaemolyticus, Bacillus subtilis, Salmonella typhimurium, Pseudomonas aeruginosa and Escherichia coli Were 18.0, 16.0, 16.0, 13.0, and 11.5mm, respectively. In particular, the antimicrobial activity of Vibrio parahaemolyticus was the strongest.

Antimicrobial Activity of Taxa Extracts Strain Methanol extract Ethanol extract Water extract Bacillus subtilis 14.0 13.5 16.0 Pseudomonas aerignosa 14.0 14.0 13.0 E.coli 11.0 11.0 11.5 Salmonella typhymurium 14.0 13.5 16.0 E.coli O-157 8.0 8.0 8.0 Vibrio parahaemolyticua 16.0 16.5 18.0 (Note) Unit: mm

As a result of investigating the effect of Alismatis Rhizoma extract on the growth of microorganisms, as shown in Figs. 3a to 3e, the growth of the bacteria was inhibited even by adding 30 μl of the Taxa extract per 5 ml of the medium. The growth of the bacteria was inhibited by more than 50% compared to the control, and the growth of the bacteria was almost inhibited when 200μl addition.

As a result of investigating the morphological changes of the microorganisms according to the extract of the taxa extract, as shown in FIG. 3, after treating the taxa water extract with Salmonerlla typhimurium , the surface structure of the cells was severely compared with the control group. Seo et al. (Seo Kwon Il, Sang Won Lee, Ki Ho Ho: Korean Agricultural Products Storage and Distribution Association (1999), 6 (1)), treated with E. coli O-157 extracts of cornus water, were observed by electron microscopy. It was reported that the cell membrane function was disrupted in the cells treated with the extract and cell contents leaked to the outside of the cells, thereby inhibiting the growth of the cells. It was reported that the structure was severely collapsed.These phenomena are also the result of the exposure of lysates or cell components due to the destruction of the cell wall and cell membrane of microorganisms by taxi water extract. I think.

Experimental Example 2: Investigation of Antioxidant Effect of Taxa Extract

The in vitro antioxidant and hepatic lipid peroxidation inhibitory effects on linoleic acid of Alismatis Rhizoma were investigated. In order to measure the antioxidant effect of taxa extract, 1 g of linoleic acid, 10 ml of ethanol, and a predetermined taxa extract were added to a Erlenmeyer flask, and 25 ml of 0.2 M phosphate buffer was added and stored at 37 ° C. for a period of time. Then, the reaction solution was transferred to a separatory funnel, and 25 ml of chloroform was added thereto, followed by extraction two to three times. 25 ml of acetic acid and 1 ml of saturated KI solution were added to the chloroform extract, which was allowed to stand in the dark for 5 minutes, and then 50 ml of distilled water was added thereto and titrated with a 1/100 N Na ₂ S ₂ O ₃ solution. Taxi extract was added methanol, ethanol, and water extract, respectively, and BHT 0.1% was added and stored at 50 ° C. for 7 days to measure peroxide value.

Lipid peroxidation inhibitory effect of Taxa extract was investigated in vitro using liver homogenate in rats. Phosphate buffer solution (phospha-

After homogenization with te buffer, pH 7.4), 1M of hydrogen peroxide (H ₂ O ₂ ), 50 mM iron sulfate (FeSO ₄ ), and 0.05 ml of taxa extract were added to the homogenate, and stored at 42 ° C. for 72 hours. thiobarbituric acid reactive substances) was measured.

As a result, as shown in FIG. 5, when the methanol extract, ethanol extract, and water extract 100 μl were added, the peroxide value was lower than that of the control, and the peroxide value of the water extract was smaller than that of the methanol and ethanol extracts, but was 0.1% BHT. A little higher than when added. In addition, as a result of measuring the peroxide value after adding the water extracts of the taxi water at the concentrations of 50, 100, 150 and 200 μl, as shown in FIG. 6, all of them were very low compared to the control, and when 150 μl was added, the peroxide was stored until 5 days storage. This was hardly produced, which was about the same as adding 100 μl of 0.1% BHT. Kwon et al. (Kwon Oh-Geun, Son Jin-Chang, Sang-Chul Kim, Sang-Cheol Kim, Pyo-Sang, Park, Seung-Woo: The Korean Society for Agricultural Products and Storage (1998), 5 (281)) showed that the antioxidant effect of the ethyl acetate fraction of linoleus methanol extract on linoleic acid was 87.3%. BHT of the comparison group was reported to be 96.2%, and Seo et al. (Seo, Kwon-Il, Sang-Won Lee, and Ki-Ho Yang: Korean Agricultural Products Storage and Distribution Association (1999), 6 (1)) showed that the extracts of linusic acid were 10, 30 and After adding 50 μl and storing at 50 ° C. for 7 days, the peroxide value was measured to be 42, 30 and 25 meq / kg compared to 187 meq / kg of the control, which showed a significant antioxidant effect. Addition of only μl showed significant antioxidant effect.

The TBA value of liver lipids in rats was 3597 μmol malondialdehyde (MDA) / g liver in the control group without the addition of tax water extract as shown in FIG. 7, and the TBA value of the test groups to which the water extract was added was 1443 μmol MDA / g liver. All showed lower anti-oxidant effect than the control.

Experimental Example 3: Determination of Immune Activity of Taxa Extract

Effects of Alismatis Rhizoma Extract on the Proliferation of Immune Cells, Effects of Macrophage Cells on Nitric Oxide Production, Proliferative Response to Homologous Antigens, Inhibition of T Cell Cytokine Secretion by Homologous Antigens, and T Cells Against Homogenous Antigens The inhibitory effect of killer cells was investigated. To separate the splenocytes of the mice, the mice (BALB / c) were sacrificed with cervical distal bone, sterilized with alcohol, placed on the dissection, and the spleen was removed by dissecting the right flank. The spleen is made into single cells using tweezers, washed three times with washing medium, and diluted in an appropriate amount of 10% FCS-RPMI 1640 medium. The isolated splenocytes were diluted in 10 ml of 10% FCS-RPMI 1640, and 40 µl anti-Thy1.2 ascites were added thereto and incubated at 4 ° C. for 60 minutes. Thereafter, the mixture was centrifuged at 4 ° C and 1200rpm for 6 minutes, diluted in 5ml of FCS-RPMI 1640, and incubated for 45 minutes in a 37 ° C water bath by adding 250µl rabbit complement. Then, centrifugation was repeated twice for 6 minutes at 4 ° C. and 1200 rpm with 10 ml washing medium, diluted in 500 μl of 10% FCS-RPMI 1640 medium, and passed through a Sephadox G-10 column. Only cells were purified. Separation of T cells was diluted in 1 ml of 10% FCS-RPMI 1640 and the splenocytes were separated into nylon wool column and incubated for 60 minutes in 37 ℃, 5% CO ₂ incubator. After incubation, the nylon wool column was washed with media preheated to 37 ° C., and only non-adherent T cells were purified and used as alloreative T cells.

Splenocytes, purely isolated B cells, or purely isolated T cells were cultured in 96 well plates with allogenic splenocytes (C57BL / 6) in 1, 10, 100, 1000 ㎍ / ml concentrations. Next, the proliferation of each cell was measured on day 2 and day 3 and day 4. Measure the cell titer96 15 μl of cell titer was added to 100 μl of the culture solution on day 2, 3, and 4 using Aqueous One Solution Cell proliferation Assay (promega), followed by incubation for 4 to 8 hours, followed by 490 nm. OD value was measured at.

To investigate the effects of taxa extract on nitric oxide production in macrophages, macrophages were treated with 1, 10, 100, and 1000 µg / ml concentrations, or when LPS and IFN-γ were added. The concentration of nitric oxide produced was measured. Stable nitrogen monoxide (NO) oxide, NO ₂ ^- (nitrite) was measured using the Griess reaction. Incubate the macrophage line (RAW264.7) under appropriate conditions for 48 hours, add 100 µl of the culture supernatant to a 96well plate, and add Greiss reagent (0.1% N -naphthyl-ethylendiamine in H ₂ O + 1% sulfanilamide in After reacting for 10 minutes by adding the same amount of 5% H ₃ PO ₄ ), the absorbance was measured at 570 nm with a microplate reader (Titertek Multiscan Plus, Finland). The concentration of nitrate was calculated by comparison with a standard curve obtained by diluting sodium nitrite from 32 μM to 0.25 μM in 2-fold.

In order to investigate the effects of taxa extract on the proliferative response by homologous antigens, cell number determination experiments were performed to induce homologous antigens. Splenocytes of mouse C57BL / 6 treated with MMC, which are target cells, and T cells (T cell of mouse BABL / c), which were isolated from splenocytes with effector cells, were cultured together for 3 days. The conditions showing the proliferative response were confirmed.

In order to determine the cause of the proliferation of T cells caused by allogeneic antigens significantly inhibited by the taxa extract, the amount of cytokines produced by T cells in response to allogeneic antigens was measured. The amount of cytokines secreted when T cells reacted was recovered after 24 hours of supernatant cultured with splenocytes and taxa, and IL-2, IL-4, IFN-γ, IL- contained in the supernatant. The amount of 10 ELISA (enzyme linked immounso-

It was measured by an rbent assay kit. At this time, the limit of measurement of cytokine concentration was 10 pg / ml.

Splenocytes were isolated from C57BL / c mice using the splenocyte separation method, diluted in 2ml of 10% FCS-RPMI medium, and 200mg of mytomycin C was added to 2mg / ml. (water bath) was treated for 25 minutes. Then, wash repeatedly with centrifuged sedimentation for 5 minutes at 4 ℃ and 1200rpm for 5 minutes with washing medium.

Used as ic splenocytes or target cells.

The ratio of target cells (allogeneic spleen cells, 1 × 10 ⁷ , 5 × 10 ⁶ / well) and effector cells (Effetor cell, T cells, 1 × 10 ⁶ / well) is 10: 1 or 5 : into a final volume of 96well round bottom plate 1 so as to 100㎕ were incubated 8 hours for 4 ℃, then centrifuged 5 minutes in the precipitation 250g 37 ℃, 5% CO ₂ incubator. However, 45 minutes before the incubation time was 8 hours, the target maximum and volume correcting control (Lysis Buffer) was diluted 10-fold and added to the cell solubility to the maximum. After incubation, centrifuged for 5 minutes at 250 g at 4 ° C. Then, 50 µl of the supernatant was transferred to a 96-well plate on a flat bottom, and 50 µl substrate mix buffer was added thereto, and the aluminum foil was covered at room temperature for 30 minutes. After the reaction, 50 μl stop soultion was added and the OD was measured at 490 nm using a microplate reader. The amount of lactate dehydrogenase (LDH) in the supernatant was measured. The degree of killer cell activity was shown. The degree of killer cell activity was calculated using the following equation.

% CYTOTOXICITY = Expermental-Effector Spontaneous-Target Spontaneous × 100 Target Maximum-Target Spontaneous

As a result of measuring the proliferation of cells by the concentration of taxa extract in the spleen cells of the mouse by concentration, as shown in FIG. 8a, the concentration of the splenocytes in the experimental group treated with the extract was increased in a concentration-dependent manner as shown in FIG. Could. As a result of measuring the induction of B cell proliferation of taxa by separating only B cells contained in splenocytes, as shown in FIG. 8B, proliferation was increased in a concentration-dependent manner as in splenocytes, especially at a concentration of 1000 ㎍ / μl. It was found that there was an excellent synergistic effect.

As a result of investigating the effect of taxa extract on nitric oxide (NO) production of macrophage, as compared to the untreated control group, as shown in Table 2, the experimental group treated with taxa extract alone by concentration and LPS were treated together with LPS. None of the experimental groups induced carbon monoxide (NO) production in macrophages. However, the experimental group treated with 1 ng / ml IFN-γ showed a tendency to decrease slightly at low concentrations of taxa extract, but significantly increased NO production at high concentrations. In other words, it can be seen that the taxa extract has a synergistic effect on the production of nitrogen monoxide (NO) induced by IFN-γ at high concentrations such as 1000 µg / µl.

Effects of Taxa Extracts on Nitric Oxide Production in Macrophage Cell RAW 264.7 Taxa Extract Stimulant 0 LPS (1 μg / ml) IFN-γ (1 ng / ml) 0 4.17 ± 1.57 3.81 ± 1.25 5.47 ± 1.74 One 2.90 ± 0.13 3.12 ± 0.08 2.86 ± 0.23 10 2.87 ± 0.24 2.66 ± 0.19 3.50 ± 0.20 100 3.02 ± 0.10 3.02 ± 0.71 6.43 ± 0.01 1000 4.49 ± 1.47 5.87 ± 2.62 21.44 ± 3.04 Taxa extract unit: µg / ml

As a result of investigating the effect of taxa extract on the proliferative response by homologous antigen, as shown in FIG. 9A, the number of effect cells was 1 × 10 ⁶ cells per well when the same number of target cells (5 × 10 ⁵ cells / well). Maximum proliferative response was seen at 3 days after culture. Therefore, as a result of measuring the proliferative response on day 3 after cultivation by treating 0.01, 0.1, 1, 10 ㎍ / ml taxa extract under the same conditions, as shown in FIG. The reaction was suppressed. In other words, the maximum inhibitory effect of about 50% at 1 ㎍ / ㎖.

As a result of measuring the amount of cytokines produced by T cells, T cells responding to homologous antigens produced IL-2, IL-10 and IFN-γ, as shown in Table 3. It was found that the production of IL-2, IFN-γ and IL-10 was suppressed in the experimental group added. Therefore, it is thought that the taxa extract inhibits cleavage proliferation by inhibiting cytokine production of T cells in response to homologous antigens.

Effects of Taxa Extracts on Cytokine Secretion of T Cells Responding to Homologous Antigens Experimental conditions Cytokines IL-2 IFN-γ IL-4 IL-10 T cell <10 94.3 ± 17.2 <10 200.0 ± 21.2 T cell + spleen 372.4 ± 4.2 644.3 ± 12.1 104 ± 13.3 430.0 ± 148.5 T Cell + Spleen + Taxa Extract 316.4 ± 3.7 422.9 ± 23.2 <10 205.0 ± 70.7 (Note) Cytokine Unit: pg / ml

As a result of investigating the effect of taxa extract on the T cell apoptosis on allogeneic antigen, as shown in FIG. 10, the activity of T cells on the homologous antigen was completely inhibited at a low concentration of 1㎍ / ml And it was found. Therefore, the extract of Taeksa inhibited the proliferation by inhibiting the cytokine production of T cells against homologous antigens, and also inhibited the killer cell activity against homologous antigens of T cells.

Example 1 Preparation of Taxa Drink

Step 1: Preparation of Taxa Extracts and Herbal Extracts

Ten times of water was added to medicinal herbs such as taxa, jujube, wolfberry, and Schisandra chinensis, respectively, and extracted with two times at 80 ° C. for 5 hours. The extract was centrifuged at 5 to 10 ° C. at 8,000 rpm for 20 minutes, filtered to obtain supernatant, and concentrated under reduced pressure to 40% of water while maintaining the temperature at 60 ° C. or less. At this time, as a result of measuring the solids yield of the concentrate, as shown in Table 4, the most number of taxis were 34.45%, followed by wolfberry, Schisandra chinensis, Jujube.

Solid Yield of Herbal Medicines Used in Taxi Beverages Yield Herb Yield (%, solids) TAX 34.45 Jujube 28.57 Wolfberry 33.33 Schisandra 31.59

Second Step: Blending Taxa Extract, Herbal Extract and Substance

First, in order to determine the drinking concentration of the taxa extract prepared in the first step, the concentration of the extract of the taxa extract to 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0%, respectively, then the herbal medicine jujube wolfberry and Sensory evaluation was conducted several times several times by adding appropriate amounts of apple juice, acidulant, sweetener, and fragrance, such as Schisandra chinensis and the ingredients, and considering the formulation suitability, manufacturing characteristics, quality characteristics, etc. Selection was made as shown in 5. Taxa extract, herbal extracts, subsidiary materials and the like were weighed and mixed according to the selected optimal blending ratio.

Ingredients and Contents of Taxa Health Drinks Content Content (g) Remarks TAX 2.25 Herbal medicine Jujube 1.50 Herbal medicine Wolfberry 0.60 Herbal medicine Schisandra 0.24 Herbal medicine Apple juice 1.31 Myeongshin Hwaseongsa Temple Citric acid 0.06 Hongsung Pharmaceutical Vit. C 0.02 Hongsung Pharmaceutical Liquid fructose 12 Doosan Mortgage 3 Samyang Genex Flavor 0.1 Hanbul Purified water Quantity system 100 ml

Third process: flavoring, dressing, and filtration

Perfume was added to 0.1 g / 100 ml of the taxi extract, the herbal extract and the adjuvant mixture prepared in the second step, and purified by adding purified water, followed by membrane filtration with a 1 μm filter.

4th process: Sterilization and filling

The filtrate prepared in the third step was sterilized at 93 ° C. for 15 seconds and then charged into a 100 ml container.

5th Process: Sterilization and Packaging

The packed sample was sterilized at 80 ° C. for 20 minutes and then packaged to produce a product as shown in FIG. 11.

As described above, the beverages are mixed with medicinal herb extracts, such as jujube, wolfberry and Schisandra chinensis, apple juice, acidulant, sweetener and flavoring agent, which have excellent antimicrobial, antioxidant and immune activities. It is a very useful invention in the food and beverage industry and national health promotion because it has the effect of producing a functional health-enhanced taxi health drink.

Claims (2)

(1) extracting and concentrating the crude herb containing taxachus, jujube, wolfberry and schizandrae at 80 ° C. for 2 hours at 80 ° C. for 5 hours to prepare a tack extract and a herbal extract; (2) herbal extracts, such as 2.0 to 3.0 g / 100ml of the tactile extract of step (1), 1.0 to 2.0g / 100ml of the extract, 0.1-1.0g / 100ml of Goji extract, and 0.1 to 0.5g / ml of Schisandra chinensis extract Weighing and mixing 1.0 to 2.0 g / 100 ml of phosphorus apple juice, 0.01 to 0.1 g / 100 ml of citric acid, 0.01 to 0.05 g / 100 ml of vitamin C, 10 to 15 g / 100 ml of liquid fructose, and 1.0 to 5.0 g / 100 ml of fructose; (3) adding 0.05 to 0.15 g / 100ml of the fragrance to the sample of step (2), adding water and allowing it to pass through, followed by membrane filtration with a 1 μm filter; (4) sterilizing the filtrate of step (3) at 93 ° C. and then filling the container; (5) Taxi health beverage manufacturing method characterized in that consisting of the step of sterilizing at 80 ℃ the sample filled in the step (4). Taxa health beverage prepared by the method of claim 1, characterized in that the pH is 3 to 4 and 12 to 13 ° Bx.