KR100770775B1 - Functional drink for immune- enhancing and Manufacturing method thereof - Google Patents

Abstract

     The present invention relates to a functional beverage for immune boosting. More specifically, red ginseng concentrate, wild ginseng cultured root concentrate, herbal medicine concentrate and chitosan as main ingredients, red ginseng concentrate 0.8-1.5% by weight, wild ginseng cultured root concentrate 0.01-0.05% by weight, and herbal medicine 0.7-1.5 (v / v) ), 0.1 ~ 0.7% by weight, hawthorn, Angelica, goji berry and licorice concentrate, vitamins, citric acid, amino acid, amino acid, amino acid, amino acid, amino acid, amino acid, It consists of mixing an appropriate amount of oligosaccharide.

The functional beverage of the present invention specifically exhibits an immune enhancing effect through cell lines and clinical experiments of animals ( in vitro, in vivo ).

Red ginseng, wild ginseng culture root, chitosan, spleen index

Description

Functional drink for immune enhancement and manufacturing method thereof

Figure 1 shows the activity of the acidic kinase of the macrophage U937 as the treatment concentration of the immuno-enhanced beverage increases.

Figure 2 shows the anti-complement effect according to the concentration of immuno-enhanced beverages.

Figure 3 shows the results of measuring the activation of macrophages in vivo conditions after the administration of immuno-enhanced beverages.

Figure 4 shows the results of measuring the activation of macrophages in vitro conditions after the treatment of immuno-enhanced beverages.

Figure 5 shows the spleen index (spleen index) indicating whether the change in the spleen after administration of the immuno-enhanced beverage.

Figure 6 shows the lymphocyte ratio in the white blood cells of the mouse blood after administration of the immuno-enhanced beverage.

The present invention relates to a functional beverage for immune boosting. More specifically, red ginseng concentrate, wild ginseng cultured root concentrate, chitosan and chitooligosaccharide as main ingredients, it is composed by mixing the appropriate amount of jujube, mountain lion, Angelica, wolfberry, licorice concentrate and yeast extract, vitamin, citric acid, amino acid, oligosaccharide.

Chitosan is a natural polymer polysaccharide extracted from shells of shellfish such as crabs and shrimps, and insects, and many kinds are produced and distributed with different molecular weights, salt types, and deacetylation degrees. Its functions are also antimicrobial, moisturizing and cohesive, and it has various functions such as fat adsorption, cholesterol excretion, antimicrobial activity, immunity enhancement, and dietary fiber function. It is applied to various fields such as field. In particular, as interest in functional foods has soared, chitosan rice, chitosan kimchi, chitosan pickled fish, or chitosan-added seaweed is produced using chitosan with various functions, and exported to the US, Canada, Australia and other countries to earn foreign currency. I'm listening. The functions of chitosan applied to such foods are known as fat adsorption, cholesterol excretion, antimicrobial activity, resiliency of macrophages to enhance immunity and dietary fiber function.

In general, chitosan is widely used in various fields such as wastewater treatment applications, agricultural applications, food and beverage applications, cosmetic applications, health and hygiene applications, textile applications, and pharmaceutical applications.

Chitosan is the only natural product that has ammonium (NH ₄ ⁺ ) ions, which is easy to bind chemically with anions, and releases harmful anions such as heavy metals, bacteria, viruses, and dioxins in the body. In addition, chitosan is known to capture fat and cholesterol and release it to the outside because it acts as a fat trapping agent in the digestive system by combining with excess fatty substances. In addition, chitosan has antimicrobial activity and defense mechanisms, which enhances the self-protection function of plants and activates plant cells due to their ability to promote germination.

On the other hand, animals' immune system is under the control of the autonomic nervous system, and when stressed, autonomic nervous system regulation is impaired in the organs under stress, causing changes in the number of immune cells in the blood, that is, the number of white blood cells. Leukocytes are largely divided into granulocytes and lymphocytes, and they perform immune functions while maintaining an appropriate ratio. Granulocytes are mainly responsible for removing large bacteria such as fungi and E. coli, and lymphocytes are responsible for removing microbes, viruses and cancer cells. However, when the body is tired or stressed, the sympathetic nerve becomes nervous and increases the number of granulocytes. Granulocytes do an important job of killing bacteria that have invaded the body, but too much can harm the body. Over-produced granulocytes kill fungi, etc. in excess, and then attach to the mucous membranes in the blood and die when free radicals are made. Too much of this free radical spreads tissue breakdown and destroys the mucous membranes of the stomach and intestines, resulting in gastric cancer, colon cancer, lung cancer, and uterine cancer. The activity of lymphocytes is suppressed, which makes them susceptible to viral diseases and cancer. If the body becomes too stable, lymphocytes increase abnormally, causing allergic diseases such as atopic dermatitis, bronchial asthma, and allergic rhinitis. Overreaction causes severe fever and inflammation even with a mild cold. Therefore, by maintaining a constant ratio of lymphocytes and granulocytes in white blood cells, it is possible to enhance immunity and prevent disease.

Ginseng is botanically belonging to the genus Araliacea and Panax ginseng, and its root is mainly used for medicinal purposes. There are six to seven species of plants belonging to the world, but three kinds of ginseng are economically cultivated and distributed as commodities in the ginseng market. Panax ginseng C.A.Meyer's botanical name, which is geographically distributed and cultivated in the Far East of Asia, has traditionally been used as the most important medicinal ingredient among Chinese herbal medicines.

Ginseng has been incredibly effective in treating various diseases and promoting energy recovery since ancient times, and continues to research extensively to explore the efficacy and pharmacological effects of ginseng. It can be said to be the homeostatic function of body control function, and based on this action, it can be anti-fatigue and anti-stress action, anti-diabetic action, blood pressure control action, anti-cancer action and brain function enhancement. It is judged that it is a result by. The main active ingredients of ginseng are known saponins, sapongenins, polyacetylenes, pyrazine derivatives, maltol, etc. In particular, the ginseng line is known to have many beneficial ingredients or contents that are not included in the ginseng line.

Although ginseng derived from wild ginseng and wild ginseng has the same origin as ginseng, ginseng has a short breeding period and has been screened for the purpose of quantitative productivity rather than effect. While it is recognized directly or indirectly, wild ginseng and wild ginseng derived from wild ginseng, which are close to their origin, have been found to maintain a complex genetic structure. In this regard, the difference between wild ginseng or wild ginseng derived from wild ginseng is not only phenomenal by oral effects but also by modern analysis methods.

Therefore, if a beverage containing chitosan and wild ginseng culture roots is easily developed and supplied to consumers, it may contribute to the improvement of national health.

On the other hand, as a prior art of the present invention, Korean Patent Publication No. 10-2004-0096153 (drinks containing chitosan curing germinated brown rice extract and a preparation method thereof) relates to a drink containing chitosan curing germinated brown rice extract, -2003-0010293 (beverage containing high molecular weight water soluble chitosan) relates to a beverage containing a high molecular weight water soluble chitosan having a weight average molecular weight of about 600,000 and is different from the constitution of the present invention.

The present invention has been proposed to solve the problems of the prior art as described above, an object of the present invention is to provide a functional drink for immuno-enhancing containing red ginseng concentrate, wild ginseng cultured muscle concentrate, chitosan as a main component.

The present invention for achieving the above object comprises a ginseng concentrate, a herbal medicine concentrate and chitosan as a main component, and includes an immune enhancing functional drink comprising a beverage excipient and purified water.

In the above, the ginseng concentrate is extracted from red ginseng and wild ginseng cultured root, characterized in that it comprises 0.8 ~ 1.5% by weight red ginseng concentrate and 0.01 ~ 0.05% by weight of wild ginseng cultured root concentrate.

In the above, the herbal medicine concentrate is extracted from jujube, hawthorn, Angelica, wolfberry and licorice, jujube concentrate 0.7-1.5% by weight, mountain lion concentration 0.1-0.7% by weight, Angelica concentrate 0.1-0.7% by weight, Goji berry concentrate 0.1-0.7% by weight , Licorice concentrate is characterized in that composed of 0.1 to 0.7% by weight.

In the above, chitosan is characterized by consisting of 0.016 to 0.064% by weight, chitooligosaccharides of 0.2 to 0.7%.

In the above, vitamins, citric acid, amino acids and oligosaccharides are further included as a beverage excipient.

In addition, the present invention extracts each component from red ginseng, wild ginseng culture root, jujube, hawthorn, Angelica, goji berry and licorice with hot water or ethanol, and then concentrated to obtain each concentrate, chitosan solution, chitooligosaccharide in each concentrate And mixing the beverage excipient and sterilizing and filtering.

Hereinafter, the present invention will be described in detail.

Functional beverage for immunological enhancement of the present invention is configured as follows. Commercially available extracts of ginseng, herbal medicine, and yeast are concentrated and used or extracted by adding hot water or ethanol to these ingredients, and then concentrated and mixed with appropriate amounts of chitosan, vitamins, citric acid, amino acids and oligosaccharides. It is composed.

The ginseng concentrate is purchased 6-year-old red ginseng in the market (Gumsan, Chungnam), and 8 times purified water is added to 1 kg of red ginseng and heated to 60-90 ° C for 12 to 24 hours to fully extract the red ginseng components. In the case of extraction with alcohol, it may be extracted with ethanol at 30 to 50 ° C. for 8 to 12 hours, or commercially available red ginseng extract may be used. Since the concentration of red ginseng extract is low, red ginseng concentrate is obtained by vacuum concentration to 1/3.

Wild ginseng cultivated root concentrates can be purchased from the market (Gyeongdong Market) or purchased from wild ginseng cultivated roots and extracted and concentrated like red ginseng concentrates.

As a herbal herb concentrate, selected jujube, mountain lion, Angelica, goji berry, and licorice can be purchased at the market (Gyeongdong Market) and extracted and concentrated like red ginseng concentrate.

The chitosan solution is obtained by dissolving the polymer chitosan in about 6.5% apple cider vinegar and using it in the form of about 2% chitosan solution.

Yeast extract powder is purchased directly from the market.

Vitamin is used by purchasing a vitamin B _1, vitamin B _2, niacin, calcium pantothenate, vitamin B ₆ on the market.

As acids, organic acids are citric acid and amino acids are L-cysteine and L-methionine. Isomaltooligosaccharides are commercially available.

Chitosan solution, chitooligosaccharides and beverage excipients were mixed with each concentrated solution of the red ginseng, wild ginseng culture root, jujube, mountain lion, Angelica, goji berry and licorice extract, and sterilized at 60-80 ° C., followed by impurities with a filter. Remove and pack.

The functional beverage of the present invention specifically exhibits an immune enhancing effect through cell lines and clinical experiments of animals ( in vitro, in vivo ).

Hereinafter, the contents of the present invention will be described in more detail with reference to Examples. However, these examples are only presented to understand the content of the present invention, but the scope of the present invention is not limited to these embodiments.

Example 1 Preparation of Beverage

For the preparation of functional beverages for immune enhancement, red ginseng concentrate, wild ginseng cultured muscle concentrate, and chitosan were prepared as shown in Table 1. Each component was completely dissolved in an appropriate amount of purified water and then charged to 100% by weight.

<Table 1> Ingredients (% by weight) of immune enhancing beverage

ingredient Sample 1 (%) Sample 2 (%) Sample 3 (%) Red ginseng concentrate One 1.2 1.5 Wild ginseng root concentrate 0.02 0.03 0.05 Jujube concentrate One One One Mountain Lion Concentrate 0.5 0.5 0.5 Angelica Concentrate 0.5 0.5 0.5 Goji Concentrate 0.5 0.5 0.5 Licorice Concentrate 0.5 0.5 0.5 Yeast extract 0.1 0.1 0.1 Salmon Eel Extract 0.03 0.03 0.03 Chitosan solution 3 2 3 Vitamin B ₁ 0.005 0.005 0.005 Vitamin B ₂ 0.005 0.005 0.005 Niacin 0.007 0.007 0.007 Calcium Pantothenate 0.005 0.005 0.005 Vitamin B ₆ 0.004 0.004 0.004 Citric acid 0.2 0.2 0.2 L-cysteine 0.01 0.01 0.01 L-Methionine 0.01 0.01 0.01 Isomaltooligosaccharide 5 5 5 Enzyme Treatment Stevia 0.05 0.05 0.05 Nahan fruit extract 0.03 0.03 0.03 Red ginseng flavor 0.01 0.01 0.01 Grapefruit Seed Extract 0.02 0.02 0.02 Sodium benzoate 0.02 0.02 0.02

Six-year-old red ginseng was purchased from the market (Gumsan, Chungnam), and three times purified water was added to 1 kg of red ginseng, and heated to 70 ° C. for 20 hours to sufficiently extract the red ginseng component. Since the concentration of red ginseng extract was low, it was used by vacuum concentration to 1/3. Wild ginseng cultivated root concentrate was purchased from the market (Gyeongdong Market) and extracted and concentrated like red ginseng concentrate.

Herbal medicine concentrates are selected jujube, hawthorn, Angelica, goji berry and licorice from the market (Gyeongdong Market), extracted and concentrated like red ginseng concentrate, and chitosan solution is obtained by dissolving it in 6.5% apple cider vinegar. It is used in the form of 2% chitosan solution.

Yeast extract was purchased directly from the market.

Vitamin B ₁ , vitamin B ₂ , niacin, calcium pantothenate, and vitamin B ₆ were purchased commercially.

As acids, citric acid and amino acid, L-cysteine and L-methionine, were used in the market.

As the sugar, isomaltooligosaccharide was purchased commercially and used.

* Mixing process and sterilization filtration process:

Chitosan solution and beverage excipients were mixed with each concentrated solution of red ginseng, wild ginseng cultured root, jujube, mountain lion, Angelica, goji berry and licorice, and sterilized at 80 ° C., and then packaged by removing impurities with a filter.

Example 2 In vitro Efficacy Verification of Immune Enhancement by Immunostimulating Beverage Using Cell Lines

The efficacy of the immunopotentiated beverage prepared in Example 1 was verified using the U937 cell line. The immune-enhanced beverage was obtained at 0, 50 µg / ml, 100 µg / ml, 150 µg / ml and 200 µg / ml concentrations for 2 hours. After treatment, the amount of lysosome according to the activity of acid kinase of U937 cell line was measured by using an acid phosphatase assay. Acid kinase activity was performed using 1M sodium acetate containing 2 mM para-nitrophenyl phosphate (PNP) and repeated three times. As the treatment concentration of the sample increases, the activity of the acidic kinase of U937 increases, and when the treatment concentration is 200 ㎍ / ㎖ increased 1.5 times (Fig. 1). In view of this, it is believed that the immune-enhanced beverage of the present invention can increase immune function through activation of macrophages.

[Embodiment 3] In the in vivo increase immune function of immune enhancing beverages using animal experiments Efficacy Verification

(1) anticomplement activity

To investigate the effects on complement activation, complement assays were performed using an antibody test called the complement fixation test, which uses the property that complement can hemolyse red blood cells in the presence of specific anti-hemolysin. The degree of activation is measured.

Normal human serum, GVB ⁺⁺ and the same amount of sample was added to the primary reaction (37 ℃, 30 minutes), diluted to 100 magnification, sensitized blood cells were added and the secondary reaction (37 ℃, 1 hour) hemolysis of sensitized blood cells The degree of inhibition is measured at absorbance 412 nm.

Anti-complement activity is an antibody test called the complement fixation test that uses the property that complement can dissolve erythrocytes in the presence of specific anti-hemolysin. The degree of activation of complement was measured.

In this experiment, Figure 2 shows the anti-complement effect according to the concentration of immuno-enhanced beverage. It was found that anti-complement activity was increased in a concentration-dependent manner, and showed maximum activity at 1000 µg / ml. Compared to the LPS (lipopolysaccharide) used as a positive control, low activity (100, 500㎍ / ㎖) showed low activity, but high concentration (1000㎍ / ㎖) showed higher activity than the positive control. As a result, it is considered that the sample has high anti-complement activity.

(2) Immune boosting effect using experimental animals

Experimental Animals:

Animals used in this experiment were male 5 week old BALB / C mice. During the experiment, water and feed were supplied in sufficient quantities, and the animal laboratory maintained a temperature of 22 ° C. and a humidity of 55% and induced a light-dark cycle at 12 hour intervals.

Preparation of Samples:

Samples were prepared using sterile physiological saline, and the dosage was 20 mg / kg mice, the control group was administered saline, when not administered it was used while stored in the refrigerator.

Activity analysis of macrophage lysosomal enzymes:

Five animals per group were injected daily intraperitoneally with a laboratory pretreated sample of 600 µl (1 mg / ml), and after 2 and 3 days, blood was obtained by migrating mice and macrophages obtained by abdominal washing. Was applied to the experiment.

Redispersed in RPMI-1640 medium so that the number of macrophage (macrophage) recovered from the mouse abdominal cavity was 1 × 10 ⁶ cells / ml, and the dispersion was dispensed in 200 μl into each well of a 96-well plate, followed by 2 at 37 ° C. Incubate for ₂ hours to fix macrophages in the wells, and then add RPMI-1640 complete medium (containing 10% FBS) to each well for 24 hours, and then add 0.1% Triton X-100 to activated macrophages. Cell membranes were lysed. At this time, the amount of phosphatase of lysosomal secreted was measured by absorbance at 405 nm with ELISA reader to indicate the degree of activity of macrophages.

The results of measuring the activation of macrophages in vivo (in vivo) are shown in FIG. 3. When the control group of physiological saline was seen as 100%, the treated group of the beverage showed an enhanced activity of macrophages of 137% (Fig. 3). As a result, the administration of the beverage seems to show a macrophage proliferation ability in vivo conditions, it is believed that this sample has an effect of enhancing the immune capacity in vivo. Saline (Saline: 0.9% NaCl) was used as a negative control.

In addition, the activation of macrophages was measured in vitro ( in vitro ) conditions. Saline (0.9% NaCl) was used as a negative control and lipopolysaccharide (LPS) was used as a positive control. As a result, it was found to be higher than the activity of lipopolysaccharide (from E. coli 026: 86, LPS), which is a cell wall component of Gram-negative bacteria used as a positive control at a concentration of 100 ㎍ / ㎖ (Fig. 4). As a result, the beverage of the present invention is considered to have a relatively strong macrophage activity.

Since macrophages have different receptors that can identify these substances, the beverage may be directly involved in the activity of macrophages, but may be due to secondary action through complement or other lymphocytes. Therefore, combining the results of the two immune activities (anti-complement and macrophage activity) of the beverage, this sample is considered to have the effect of boosting the immune system. However, it is not clear at this time which direction their immune-boosting effects will progress in the two interrelated immune systems.

Spleen index was used as an indicator of the change in the spleen after treatment. This indicator is expressed by dividing the spleen weight by the total weight of the animal and multiplying by 100 to express the change in the weight of the spleen. As a result of measuring the spleen indices of the mice treated with the beverage, it showed an increase of 8% compared to the saline group (FIG. 5). It is believed that the beverage has a function of increasing splenocyte proliferation.

Table 2: Number of Spleen-Derived Cells and Macrophages

Group Splenocyte number (10 ⁷ ) Macrophage number (10 ⁷ ) 2 D 3 D 2 D 3 D Saline 101 131 120 205 Beverage 120 110 201 380

20 mg / kg were administered to the mouse abdominal cavity.

In addition, splenic-derived cells and macrophages were measured after 3 days of intraperitoneal administration of immune-enhanced beverages. However, splenocytes showed no significant change, but macrophages showed a sharp increase in cell numbers.

In addition, lymphocyte counts were measured in the blood after 3 days intraperitoneal administration of immunopotentiated beverages. As a result, as shown in Figure 6, the immune-enhanced beverage-administered group in the ratio of lymphocytes in the white blood cells is higher than the physiological saline-administered group, since the number of lymphocytes that regulate immunity, it can be judged that the immune regulation is as high (Fig. 6 □; Physiological saline, immunity-enhanced beverages (20 mg / kg body weight).

As described above, the present invention is a functional beverage exhibiting a specific immunity-enhancing effect through clinical experiments of cell lines and experimental animals, and is a good drink for stressful office workers, middle-aged men and women, elderly people with weak physical condition, and examinees. Can provide.

Claims (6)

Extracts from red ginseng and wild ginseng cultured roots; Extracted from jujube, hawthorn, Angelica, wolfberry and licorice. Jujube concentrate 0.7-1.5% by weight, mountain lion concentration 0.1-0.7% by weight, Angelica concentrate 0.1-0.7% by weight, wolfberry concentrate 0.1-0.7% by weight, licorice concentrate 0.1-0.7 weight%; 0.016 to 0.064 weight% chitosan and 0.2 to 0.7 weight% chitooligosaccharides; Beverage excipients include vitamins, citric acid, amino acids and oligosaccharides; And purified water, characterized in that the functional beverage for immuno-enhancement. delete delete delete delete delete

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