KR102537844B1 - Composition with antibacterial, antioxidant, and anti-inflammatory activity containing ingredients converted to kimchi lactic acid bacteria and breast lactic acid bacteria in marigold extract - Google Patents

Abstract

The present invention relates to a composition having antibacterial, antioxidant and anti-inflammatory activities containing a component obtained by bioconversion of marigold extract to breast milk lactic acid bacteria, and marigold ( Tagetes erecta ) cultured extract of breast milk lactic acid bacteria according to the present invention is antioxidant, It has remarkably excellent anti-inflammatory and antibacterial effects.

Description

Composition with antibacterial, antioxidant, and anti-inflammatory activity containing ingredients converted to kimchi lactic acid bacteria and breast lactic acid bacteria in marigold extract}

The present invention relates to a composition having antibacterial, antioxidant and anti-inflammatory activities containing a component obtained by bioconversion of marigold ( Tagetes erecta) extract into breast milk lactic acid bacteria.

Reactive oxygen species generated during the metabolism of the human body cause damage to cell membranes, proteins, and DNA in the body, which can cause aging and disease. Free radicals and other reactive oxygen species (ROS) can be produced as by-products during oxidation in living organisms (Halliwell & Gutteridge, 1999). Reactive oxygen species (ROS) play many important functions in normal metabolism, but excessive ROS can impair protein function, cause oxidative damage to cellular macromolecules, and trigger cell death (Kremer et al. 2012 ). Long-term conditions of oxidative stress are involved in the pathogenesis of several diseases including cardiovascular disease and cancer (Sun et al. 2004). Oxidative damage to DNA, proteins and other macromolecules is further hypothesized to constitute the major type of intrinsic damage leading to aging (Fraga et al. 1990). Therefore, as interest in enjoying a healthy life by ingesting antioxidants increases, interest and demand for natural antioxidants that are harmless to the human body are increasing, and various antioxidants derived from natural products are being explored (Choi et al ., 2005).

Inflammation is caused by macrophages in the human body responding to external stimuli or invading pathogens to produce cytokines such as TNF-α, interleukin1-β (IL-1β) and IL-6, which are inflammation-inducing factors, and also inducible nitric oxide. Synthase (iNOS) and cyclooxygenase-2 (COX-2) are synthesized to produce nitric oxide (NO) and prostaglandin E2 (PGE2). NO generated through this process plays a role such as removing tumors and bacteria, but when NO is excessively produced in the body, it causes tissue damage, gene mutation, and increases vascular permeability, causing inflammatory reactions such as edema. It causes damage to the human body (Moncada et al ., 1991; Choi et. al ., 1993). As a result, interest in anti-inflammatory substances is increasing.

Lactic acid bacteria are microorganisms that produce lactic acid, a type of organic acid, and are widely distributed not only in humans but also in animals and plants. In ancient dietary life, lactic acid bacteria have been widely used as a fermented species of fermented foods for the purpose of enhancing the taste and aroma of foods. Bacteriocins, which are biosynthesized in the ribosome of microorganisms, are mainly composed of proteins (peptides or proteins), and attach to the cell surface of pathogenic bacteria including Bacillus cereus, known as food poisoning bacteria, and inhibit cell wall synthesis or protein synthesis. It plays a role in killing bacterial cells (Jack, R. W. (1995) Bacteriocins of grampositive bacteria. Microbiol). Bacteriocin with these characteristics is a natural substance and has the biggest advantage of being harmless to the human body because it is made of protein. In addition, through industrialization processes such as genetic manipulation and optimization of culture conditions, more antibacterial substances can be produced through smooth culture of strains and mass production.

Marigold ( Tagetes erecta) , the material of this study, is an annual plant of the Asteraceae family, which is called marigold in Korea, and is native to Southern Europe. Marigolds are known to be effective in inflammatory diseases caused by edema caused by saponins, which are main components, and have antibacterial effects. Recently, studies have been conducted to increase the physiological activity of raw materials by fermentation technology using beneficial microorganisms such as molds, yeasts, lactic acid bacteria and mushrooms.

Along with the development of science and technology, many studies and developments have been made on microorganisms, among which lactic acid bacteria are known to produce bacteriocin, which is known as a low-molecular antibacterial substance. Fermentation by lactic acid bacteria is an old food processing method that imparts a variety of flavors and textures to food, increases the value of food by producing nutritional and functional ingredients, and improves shelf life by biosynthesis of antimicrobial substances.

Kimchi lactic acid bacteria (Lactobacillus plantarum) is well known as a lactic acid bacterium that produces a bacteriocin called plantaricin. Lactobacillus gasseri is known to exist in abundance in the human gastrointestinal tract. According to some previous studies, fermentation products of L. gasseri are known to have effects on immune regulation, antibacterial and antihypertensive agents, but poor growth activity of L. gasseri in milk has been reported.

Based on this report of various physiological and chemical functions, each marigold extract bioconversion product was prepared using breast milk lactobacillus and kimchi lactobacillus, phenol and flavonoid contents were evaluated, and various antioxidant and antibacterial efficacies were compared and analyzed. reviewed.

Registered Patent Publication No. 10-2035261

An object of the present invention is to provide an anti-aging composition comprising a breast milk lactic acid bacteria cultured extract of marigold ( Tagetes erecta ) extract.

Another object of the present invention is marigold ( Tagetes erecta ) To provide an anti-inflammatory composition comprising a breast milk lactic acid bacteria culture extract of the extract.

Another object of the present invention is marigold ( Tagetes erecta ) To provide an antibacterial composition comprising a breast milk lactic acid bacteria culture extract of the extract.

In order to achieve the above purpose,

The present invention marigold ( Tagetes erecta ) Provides an antioxidant composition containing the breast milk lactic acid bacteria cultured extract of the extract.

The breast milk lactic acid bacteria may use Lactobacillus gasseri .

The antioxidant composition according to the present invention can be used as a health food, cosmetic or pharmaceutical raw material.

In addition, the present invention marigold ( Tagetes erecta ) Provides an anti-inflammatory composition comprising a breast milk lactic acid bacteria culture extract of the extract.

The breast milk lactic acid bacteria may use Lactobacillus gasseri .

The anti-inflammatory composition according to the present invention can be used as a health food, cosmetic or pharmaceutical raw material.

Furthermore, the present invention marigold ( Tagetes erecta ) Provides an antibacterial composition comprising a breast milk lactic acid bacteria culture extract of the extract.

The breast milk lactic acid bacteria may use Lactobacillus gasseri .

The antibacterial composition according to the present invention can be used as a health food, cosmetic or pharmaceutical raw material.

Marigold according to the present invention ( Tagetes erecta ) Breast milk lactobacillus cultured extract of the extract has a remarkably excellent antioxidant, anti-inflammatory and antibacterial effect.

1 shows the reducing power of marigold extract (Preparation Example 1), kimchi lactic acid bacteria bioconverted marigold extract (Comparative Example 1), breast milk lactic acid bacteria bioconverted marigold extract (Example 1) and ascorbic acid (AA, positive control). This is an evaluated graph.
2 is a marigold extract (Preparation Example 1), kimchi lactic acid bacteria bioconverted marigold extract (Comparative Example 1), and breast milk lactic acid bacteria bioconverted marigold extract (Example 1) by oxidative damage of superhelical DNA according to treatment This is the result of quantitative evaluation of the degree of conversion to linear DNA. (C: Control, NC: Hydrogen peroxide treatment group, 0.25 ~ 1 mg/mL: Hydrogen peroxide and sample treatment group at the same time)
Figure 3 is an image confirming the antibacterial activity against S. aureus and E. coli according to the treatment concentration of marigold extract (Preparation Example 1) and breast milk lactic acid bacteria bioconverted marigold extract (Example 1) by disk diffusion method.

Health food and health functional food composition

There is no particular restriction on the type of food. Examples of foods to which the active substance of the present invention can be added are dairy products including drinks, meat, sausages, bread, biscuits, rice cakes, chocolates, candies, snacks, confectionery, pizza, ramen, other noodles, chewing gum, ice cream, There are various soups, beverages, alcoholic beverages and vitamin complexes, dairy products and milk-processed products, etc., and includes both health foods and health functional foods in a conventional sense.

Health food and health functional food compositions containing active substances according to the present invention may be added to food as it is or used together with other foods or food ingredients, and may be appropriately used according to conventional methods. The mixing amount of the active substance may be appropriately determined according to its purpose of use (for prevention or improvement). In general, the amount of the composition in health food and health functional food may be added in an amount of 0.1 to 90 parts by weight based on the total weight of food. However, in the case of long-term intake for the purpose of health maintenance or health control, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount greater than the above range.

The health food and health functional food composition of the present invention is not particularly limited in other ingredients except for containing the active substance of the present invention as an essential ingredient in the indicated ratio, and various flavoring agents or natural carbohydrates are added as additional ingredients like conventional beverages. may contain Examples of the aforementioned natural carbohydrates include monosaccharides such as glucose, fructose, and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrins, cyclodextrins, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (thaumatin, stevia extract (eg rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can advantageously be used. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 of the health functional food composition of the present invention.

In addition to the above, the health food and health functional food composition containing the active substances of the present invention are various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate, etc.) ), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, and the like. In addition, the health food and health functional food composition of the present invention may contain fruit flesh for preparing natural fruit juice, fruit juice beverages, and vegetable beverages.

These components may be used independently or in combination. The ratio of these additives is not so critical, but is generally selected in the range of 0.1 to about 20 parts by weight per 100 parts by weight of the health food and health functional food composition containing the active substance of the present invention.

cosmetic composition

The present invention provides a cosmetic composition containing an active substance.

The cosmetic composition is, for example, a solution, gel, solid or kneaded anhydrous product, emulsion obtained by dispersing an oil phase in an aqueous phase, suspension, microemulsion, microcapsule, microgranule or ionic (liposome), nonionic vesicle dispersant It may be provided in the form of a cream, toner, lotion, powder, ointment, spray or conceal stick. In addition, it can be prepared in the form of a foam (foam) or the form of an aerosol composition further containing a compressed propellant.

In addition, the cosmetic composition includes a fatty substance, an organic solvent, a solubilizing agent, a thickening agent and a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, a surfactant, Water, ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, humectants, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or commonly used in cosmetics It may contain adjuvants commonly used in the cosmetic field, such as any other ingredients that are

In the cosmetic composition containing the active material of the present invention, the active material of the present invention may be added in an amount of 0.1 to 50% by weight, preferably 1 to 10% by weight, in the cosmetic composition that is usually contained.

When the active substance of the present invention is used as an external skin preparation, additionally, a fatty substance, an organic solvent, a solubilizing agent, a thickening agent and a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, and a surfactant , water, ionic or nonionic emulsifiers, fillers, sequestering agents and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles, or external preparations for skin It may contain adjuvants commonly used in the field of dermatology, such as any other ingredients used as In addition, the components may be introduced in an amount generally used in the field of skin science.

pharmaceutical composition

The active substance of the present invention can be administered in various oral and parenteral formulations during clinical administration, and when formulated, diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants are used. It is manufactured by

Solid preparations for oral administration include tablets, patients, powders, granules, capsules, troches, etc., and these solid preparations contain one or more active substances of the present invention and at least one excipient such as starch, calcium carbonate, It is prepared by mixing sucrose, lactose or gelatin. In addition to simple excipients, lubricants such as magnesium styrate and talc are also used. Liquid formulations for oral administration include suspensions, internal solutions, emulsions, or syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, aromatics, and preservatives may be included. can

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspension solutions, emulsions, freeze-dried formulations, suppositories, and the like. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspending agents. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerol, gelatin, and the like may be used.

In addition, the effective dosage of the active substance of the present invention for the human body may vary depending on the patient's age, weight, sex, dosage form, health condition and disease degree, and is generally about 0.001-100 mg/kg/day, Preferably it is 0.01-35 mg/kg/day. Based on an adult patient weighing 70 kg, it is generally 0.07-7000 mg/day, preferably 0.7-2500 mg/day, and once or twice a day at regular intervals according to the judgment of a doctor or pharmacist. It may be divided into several doses.

Hereinafter, the present invention will be described in more detail by the following examples. However, the following examples are merely illustrative of the present invention, and the contents of the present invention are not limited by the following examples.

<Preparation Example 1> Marigold ( Tagetes erecta ) Preparation of extract

After extracting 100 g of marigold ( Tagetes erecta ) raw material purchased from CK Co., Ltd. in 70% EtoH, the extract was filtered. Afterwards, the marigold extract was concentrated under reduced pressure at 50° C. using a vacuum concentrator to prepare a marigold concentrate.

<Preparation Example 2> Preparation of human milk lactic acid bacteria extract

KCTC 3144 ( Lactobacillus gasseri ), a breast milk lactic acid bacterium, was distributed from the Biological Resource Center KCTC, spread on MRS Agar medium, grown at 37℃ for 24 hours, picked colonies, confirmed the growth curve, and grew the bacteria on MRS medium. Sufficiently grown breastmilk lactic acid bacteria were centrifuged at 1400 rpm and 4° C. for 20 minutes to obtain only the supernatant, which was filtered with a 0.45 μL filter and lyophilized to prepare Lactobacillus gasseri strain extract.

<Example 1> Preparation of marigold extract bioconverted to breast milk lactic acid bacteria

100 g of marigold raw material purchased from CK Co., Ltd. was divided and extracted three times with 70% EtOH 12L, and the extract was filtered. Thereafter, KCTC 3144 ( Lactobacillus gasseri ) strain as breast milk lactic acid bacteria cultured in marigold extract concentrated under reduced pressure at 50 ° C using a reduced pressure concentrator, Difco Lactobacillus MRS Broth 250 mL, and Agr medium in an incubator at 37 ° C for 24 hours. Inoculated by 2% (v / v) of 500mL of marigold concentrate and cultured in an incubator for 24 hours to obtain a marigold breast milk lactic acid bacteria bioconversion product, which was freeze-dried and used as a sample.

<Comparative Example 1> Preparation of marigold extract bioconverted with kimchi lactic acid bacteria

100 g of marigold raw material purchased from CK Co., Ltd. was divided and extracted three times with 70% EtOH 12L, and the extract was filtered. Thereafter, KCCM 12116 ( Lactobacillus plantarum ) strain as kimchi lactic acid bacteria cultured in marigold extract concentrated under reduced pressure at 50 ° C using a vacuum concentrator, Difco Lactobacillus MRS Broth 250 mL, and Agr medium in an incubator at 37 ° C for 24 hours. Inoculated by 2% (v / v) of 500mL of marigold concentrate and cultured in an incubator for 24 hours to obtain a marigold kimchi lactic acid bacteria bioconversion product, which was freeze-dried and used as a sample.

<Experimental Example 1> Antioxidant evaluation

<Experimental Example 1-1> Evaluation of polyphenol and flavonoid content

Marigold extract (Preparation Example 1), kimchi lactic acid bacteria bioconversion marigold extract (Comparative Example 1), breast milk lactic acid bacteria bioconversion marigold extract (Example 1), Lactobacillus gasseri extract (Preparation Example 2) flavonoid and polyphenol content, respectively was evaluated, and the results are shown in Table 1 below.

sample flavonoid content
(μg/ CE mg) polyphenol content
(μg/GAE mg) Marigold extract (Preparation Example 1) 175.21 ± 6.14 173.46 ± 1.83 Kimchi lactic acid bacteria bioconverted marigold extract (Comparative Example 1) 186.43 ± 5.11 175.29 ± 0.80 Breast milk lactobacillus bioconverted marigold extract (Example 1) 210.54 ± 5.28 234.87 ± 3.25 Breast milk lactic acid bacteria extract (Preparation Example 2) 12.84 ± 0.06 128.21 ± 3.54

As shown in Table 1, the flavonoid and polyphenol contents of the breast milk lactic acid bacteria bioconverted marigold extract (Example 1) were found in marigold alone extract (Preparation Example 1), breast milk lactic acid bacteria alone extract (Preparation Example 2) and Kimchi Lactobacillus bioconversion It was confirmed that it appeared significantly higher than the marigold extract (Comparative Example 1).

<Experimental Example 1-2> Evaluation of reducing power

Marigold extract (Preparation Example 1), kimchi lactic acid bacteria bioconverted marigold extract (Comparative Example 1), human milk lactic acid bacteria bioconverted marigold extract (Example 1), and ascorbic acid (AA, positive control) were each evaluated for their reducing power.

Specifically, after mixing 1 ml of a sample or standard reagent with a concentration of 0.25-4 mg/mL, 2.5 ml of 1 ml of Sodium Phosphate buffer (PBS) and 1% Potassium Ferricyanide, respectively, stand at 50 ° C for 20 minutes and then react After picking up 5 ml of each of the finished mixtures and transferring them to a new tube, 2.5 ml of trichloroacetic acid (TCA) was added and centrifuged at 3,000 rpm for 10 minutes in a centrifuge. After adding 5 ml of DW and 1 ml of ferric chloride to 5 ml of the centrifuged supernatant, dispense 0.15 ml each into a 96-well plate and measure the absorbance at 700 nm using an ELISA reader. The effective concentration of each sample having an absorbance value of 0.5 (EC50) was evaluated, and the results are shown in FIG. 1 .

1 shows the reducing power of marigold extract (Preparation Example 1), kimchi lactic acid bacteria bioconverted marigold extract (Comparative Example 1), breast milk lactic acid bacteria bioconverted marigold extract (Example 1) and ascorbic acid (AA, positive control). This is an evaluated graph.

As shown in Figure 1, it was confirmed that the reducing power of the breast milk lactic acid bacteria bioconverted marigold extract (Example 1) was significantly improved compared to the marigold extract (Preparation Example 1) and the kimchi lactic acid bacteria bioconverted marigold extract (Comparative Example 1). .

<Experimental Example 1-3> Evaluation of oxidative DNA damage prevention efficacy

Prevention of oxidative damage to DNA by hydrogen peroxide (H ₂ O ₂ ) was carried out by slightly modifying the experimental method of Tian & Hua (2005). 1 μL of plasmid PBR 322 DNA (0.5 μg/μL) was mixed with 3 μL (0.08 mm) of FeSO ₄ , 4 μL (v/v) of 30% H ₂ O ₂ in 3 μL distilled water at concentrations of 0.25, 0.5 and 1 mg/mL. A mixture of 2 μL of the extract was reacted at 37° C. for 1 hour. After that, 2 μL of 6X DNA loading dye was added. It was reacted with 0.8% agarose gel electrophoresis at 70V at room temperature. DNA bands (supercoiled, linear and open circular) were stained with ethidium bromide, gels were scanned using the Gel documentation system (Nextep, Korea), and band quantification was performed using NEXTEP analysis software. The effect of inhibiting DNA oxidation was evaluated by the increase and decrease of supercoiled monomer based on the control value.

2 is a marigold extract (Preparation Example 1), kimchi lactic acid bacteria bioconverted marigold extract (Comparative Example 1), and breast milk lactic acid bacteria bioconverted marigold extract (Example 1) by oxidative damage of superhelical DNA according to treatment This is the result of quantitative evaluation of the degree of conversion to linear DNA. (C: Control, NC: Hydrogen peroxide treatment group, 0.25 ~ 1 mg/mL: Hydrogen peroxide and sample treatment group at the same time)

As shown in FIG. 2, marigold extract (Preparation Example 1), kimchi lactic acid bacteria bioconverted marigold extract (Comparative Example 1), and breast milk lactic acid bacteria bioconverted marigold extract (Example 1) all have oxidative damage to superhelical DNA. It was confirmed that there is an effect of inhibiting conversion to linear DNA by. In particular, it was confirmed that Example 1 was superior in the efficacy of protecting oxidative DNA damage compared to Preparation Example 1 and Comparative Example 1.

<Experimental Example 2> Anti-inflammatory evaluation through NO (Nitric oxide) production inhibition ability

NO (Nitric oxide) is an inorganic substance produced from L-arginine by NO synthase, which activates cells due to cytokine stimulation or microbial invasion. It is known that a large amount of NO is formed in RAW264.7 macrophages by LPS stimulation, and the resulting cytotoxicity is also involved in inflammatory reactions, cell mutations, and tumor development.

In order to evaluate the anti-inflammatory efficacy, marigold extract (Preparation Example 1), kimchi lactic acid bacteria bioconverted marigold extract (Comparative Example 1), breast milk lactic acid bacteria bioconverted marigold extract (Example 1) and breast milk lactic acid bacteria extract (Preparation Example 2 ) was evaluated for each treatment concentration.

Specifically, RAW264.7 macrophages were seeded in a 6 well plate at 1×10 ⁶ cells/well, and after 18 hours, each sample was treated with 0.25, 0.5, 1, 2, or 4 mg/mL, followed by LPS (100 mg/mL). ng/mL) and cultured for 24 hours. Then, 150 μL of cell culture medium and 150 μL of Griess reagent were mixed and reacted in an incubator at 37 ° C for 10 minutes. Then, absorbance was measured at 570 nm using an ELISA reader (BioTek, Winooski, VT, USA), and A standard curve was prepared to calculate the NO content.

NO production amount (μM) Sample treatment concentration (mg/mL) 0.25 0.5 One 2 4 Control 18.2 - - - - - LPS 100 - - - - - Example 1 - 87 80 70 58 43 Comparative Example 1 - 92 89 83 74 62 Preparation Example 1 - 96 92 88 81 71 Preparation Example 2 - 97 95 90 83 74

As shown in Table 2, NO production of the breast milk lactic acid bacteria bioconverted marigold extract (Example 1) is marigold alone extract (Preparation Example 1), breast milk lactic acid bacteria alone extract (Preparation Example 2) and Kimchi Lactobacillus bioconverted marigold extract (Comparative Example 1) appeared significantly lower than, it was confirmed that the anti-inflammatory efficacy is the most excellent.

<Experimental Example 3> Evaluation of antibacterial efficacy

In order to examine the antibacterial effect of the marigold extract (Preparation Example 1) and the bioconverted marigold extract (Example 1) of human milk lactic acid bacteria, a disk diffusion method was performed.

Specifically, S. aureus and E. coli were inoculated in advance on a plate medium, respectively, and then the circular filter paper was added to the extracts by concentration (10, 25, 50, 100, 200 mg/mL) of Preparation Example 1 and Example 1. After soaking, take them out and place them on a plate medium. After culturing, a growth-inhibited zone in which microorganisms do not grow appears in a circular shape around the disk containing the antibacterial substance.

Figure 3 is an image confirming the antibacterial activity against S. aureus and E. coli according to the treatment concentration of marigold extract (Preparation Example 1) and breast milk lactic acid bacteria bioconverted marigold extract (Example 1) by disk diffusion method.

As shown in FIG. 3, in the case of E.coli, similar antibacterial effects were observed in Preparation Example 1 and Example 1, and in the case of S. aureus, it was confirmed that the antibacterial efficacy of Example 1 was significantly superior to Preparation Example 1.

Production example of health food

The active substance according to the present invention can be manufactured into various types of health food depending on the purpose. The following exemplifies methods for producing some health foods containing the active substances according to the present invention as active ingredients, but the present invention is not limited thereto.

<Health food manufacturing example 1> Manufacturing of dairy products

0.01-1 part by weight of the active substance of the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

<Health food manufacturing example 2> Manufacturing of food

Brown rice, barley, glutinous rice, and adlay were alphanized by a known method, dried, roasted, and then prepared into a powder having a particle size of 60 mesh using a grinder. Black beans, black sesame seeds, and perilla seeds were also steamed and dried by a known method, roasted, and then prepared into powder with a particle size of 60 mesh by a grinder. The active substance of the present invention was concentrated under reduced pressure in a vacuum concentrator to obtain a dry powder. It was prepared by blending the dry powder of grains, seeds and active substances prepared above in the following ratio.

Cereals (brown rice 34 parts by weight, adlay 19 parts by weight, barley 20 parts by weight),

Seeds (7 parts by weight of perilla seeds, 8 parts by weight of black beans, 7 parts by weight of black sesame seeds),

Active substance (2 parts by weight),

reishi (1.5 parts by weight), and

Rehmannia glutinosa (1.5 parts by weight).

Production example of health functional food

The active substance according to the present invention can be manufactured into various types of health functional food depending on the purpose. The following exemplifies methods for producing some health functional foods containing the active substances according to the present invention as active ingredients, but the present invention is not limited thereto.

<Health functional food manufacturing example 1> Manufacturing of health functional food

Active substance 100 mg

Appropriate amount of vitamin mixture

Vitamin A Acetate 70 μg

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

Vitamin B12 0.2 μg

Vitamin C 10 mg

Biotin 10 μg

Nicotinamide 1.7 mg

Folic acid 50 μg

Calcium Pantothenate 0.5 mg

Appropriate amount of mineral mixture

Ferrous sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium Carbonate 25.3 mg

Potassium Phosphate Monobasic 15 mg

Dibasic Calcium Phosphate 55 mg

Potassium citrate 90 mg

Calcium Carbonate 100 mg

Magnesium Chloride 24.8 mg

Although the composition ratio of the above vitamin and mineral mixture was prepared by mixing ingredients suitable for relatively health functional foods in a preferred embodiment, the mixing ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health functional food manufacturing method. Then, granules can be prepared and used in the preparation of health functional food compositions according to conventional methods.

<Health functional food manufacturing example 2> Manufacturing of health functional beverages

Active substance 100 mg

Citric Acid 100 mg

Oligosaccharides 100 mg

Plum concentrate 2 mg

Taurine 100 mg

Add purified water to total 500 mL

After mixing the above components according to the usual health drink manufacturing method, stirring and heating at 85 ° C. for about 1 hour, the resulting solution is filtered and obtained in a sterilized container, sealed and sterilized, and then refrigerated according to the present invention. It is used in the manufacture of health beverage compositions. Although the composition ratio is a mixture of ingredients suitable for a relatively favorite beverage in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as the class of demand, the country of demand, and the purpose of use.

Production example of cosmetics

The active substance according to the present invention can be manufactured into various types of cosmetics depending on the purpose. The following exemplifies a method for preparing some cosmetics containing the active substance according to the present invention as an active ingredient, but the present invention is not limited thereto.

<Cosmetics Production Example 1> Production of softened lotion

Active substance 10.0 parts by weight

1,3-butylene glycol 1.00 parts by weight

Disodium EDTA 0.05 parts by weight

Allantoin 0.10 part by weight

Dipotassium glycyrrhizate 0.05 parts by weight

Citric acid 0.01 part by weight

Sodium citrate 0.02 parts by weight

Glycereth-26 1.00 parts by weight

Arbutin 2.00 parts by weight

Hydrogenated castor oil 1.00 parts by weight

30.0 parts by weight of ethanol

trace amounts of preservatives

trace amount of colorant

traces of flavoring

Remaining amount of purified water

<Cosmetic preparation example 2> Manufacture of nutrient cream

Active substance 10.0 parts by weight

1,3-butylene glycol 7.00 parts by weight

Glycerin 1.00 parts by weight

D-panthenol 0.10 part by weight

3.20 parts by weight of plant extract

Magnesium aluminum silicate 0.30 part by weight

1.20 parts by weight of PEG-40 stearate

2.00 parts by weight of stearic acid

Polysorbate 60 1.50 parts by weight

2.00 parts by weight of lipophilic glyceryl stearate

1.50 parts by weight of sorbitan sesquioleate

Cetearyl alcohol 3.00 parts by weight

Mineral oil 4.00 parts by weight

3.80 parts by weight of squalane

Carrylic/capric triglyceride 2.80 parts by weight

Vegetable oil 1.80 parts by weight

Dimethicone 0.40 parts by weight

Diposium glycyrrhizate trace amount

trace amounts of allantoin

trace amounts of sodium hyaluronate

Dosage of Tocopheryl Acetate

Triethanolamine dosage

appropriate amount of preservative

Appropriate amount of flavoring agent

Remaining amount of purified water

Manufacturing example of drug

The active substance according to the present invention can be formulated in various forms depending on the purpose. The following exemplifies some formulation methods containing the active substance according to the present invention as an active ingredient, but the present invention is not limited thereto.

<Pharmaceutical Preparation Example 1> Preparation of powder

2 g of active substance

1 g lactose

After mixing the above ingredients, the powder was prepared by filling in an airtight bag.

<Pharmaceutical Preparation Example 2> Preparation of tablets

Active substance 100 mg

Corn Starch 100 mg

100mg milk sugar

Magnesium stearate 2 mg

After mixing the above components, tablets were prepared by tableting according to a conventional tablet manufacturing method.

<Pharmaceutical Preparation Example 3> Preparation of capsules

Active substance 100 mg

Corn Starch 100 mg

100mg milk sugar

Magnesium stearate 2 mg

After mixing the above ingredients, capsules were prepared by filling gelatin capsules according to a conventional capsule preparation method.

<Pharmaceutical Preparation Example 4> Preparation of Injections

Active substance 10 μg/ml

dilute hydrochloric acid BP until pH 3.5

Sodium Chloride BP for Injection up to 1 ml

The active substance according to the present invention was dissolved in an appropriate volume of sodium chloride BP for injection, the pH of the resulting solution was adjusted to pH 3.5 with dilute hydrochloric acid BP, the volume was adjusted with sodium chloride BP for injection, and mixed thoroughly. . The solution was filled into a 5 ml type I ampoule made of transparent glass, sealed under an upper grid of air by dissolving the glass, and sterilized by autoclaving at 120° C. for 15 minutes or longer to prepare an injection solution.

<Pharmaceutical Preparation Example 5> Preparation of Nasal Spray

Active substance 1.0 g

Sodium Acetate 0.3 g

Methylparaben 0.1 g

Propylparaben 0.02 g

proper amount of sodium chloride

HCl or NaOH pH adjustment suitable amount

Appropriate amount of purified water

According to the conventional method for preparing nasal absorbents, saline (0.9% NaCl, w/v, solvent is purified water) is prepared to contain 3 mg of active substance per 1 mL, filled in an opaque spray container and sterilized to prepare nasal absorbents. manufactured.

<Pharmaceutical Preparation Example 6> Preparation of liquid formulation

Active substance 100 mg

Isomerized sugar 10 g

5 g mannitol

Appropriate amount of purified water

According to the conventional method for preparing liquids, each component is dissolved in purified water, lemon flavor is added, the above components are mixed, and purified water is added to adjust the total volume to 100 mL, and then filled in a brown bottle and sterilized to prepare a liquid. did

So far, the present invention has been looked at with respect to its preferred embodiments. Those skilled in the art to which the present invention pertains will be able to understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered from an illustrative rather than a limiting point of view. The scope of the present invention is shown in particular in the claims rather than the foregoing description, and all differences within the equivalent range will be construed as being included in the present invention.

Claims (9)

Marigold ( Tagetes erecta ) Extract of Lactobacillus gasseri ( Lactobacillus gasseri ) An antioxidant composition comprising a culture extract.
delete According to claim 1,
The antioxidant composition is characterized in that used as a health food, cosmetic or pharmaceutical raw material, antioxidant composition.
Marigold ( Tagetes erecta ) Lactobacillus gasseri of the extract ( Lactobacillus gasseri ) Anti-inflammatory composition comprising a culture extract.
delete According to claim 4,
The anti-inflammatory composition is characterized in that used as a health food, cosmetic or pharmaceutical raw material, anti-inflammatory composition.
Marigold ( Tagetes erecta ) Extract of Lactobacillus gasseri ( Lactobacillus gasseri ) An antibacterial composition comprising a culture extract.
delete According to claim 7,
The antibacterial composition is characterized in that used as a health food, cosmetic or pharmaceutical raw material, antibacterial composition.

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