KR20180037650A - A composition for producing metal nanoparticles comprising black ginseng extracts and the use thereof - Google Patents

Abstract

The present invention relates to a composition for manufacturing metal nanoparticles comprising an extract of Black Ginseng as an active ingredient, a method for manufacturing metal nanoparticles using the composition, metal nanoparticles manufacture by the method, and antimicrobial and antioxidant uses of the metal nanoparticles. Using the method for manufacturing metal nanoparticles of the present invention, metal nanoparticles of uniform sizes can be manufactured without adding any reducing agent or stabilizer in a short time. Moreover, since the metal nanoparticles manufactured by the method have antimicrobial activity and antioxidant activity, the composition comprising the same can be usefully used in industry.

Description

TECHNICAL FIELD The present invention relates to a composition for preparing metal nanoparticles containing black ginseng extract and a use thereof,

The present invention relates to a composition for the preparation of metal nanoparticles comprising black ginseng extract and a use thereof. More specifically, the present invention relates to a composition for the preparation of metal nanoparticles comprising black ginseng extract as an active ingredient, Methods, metal nanoparticles produced by the method, and antimicrobial and antioxidant uses of the metal nanoparticles.

Metal nanoparticles have attracted much attention in various scientific fields, especially in life sciences and engineering fields, due to their specific and specific features such as uniform particle size distribution, easy surface modification, and excellent stability (Mody VV, J. Pharm. Bioall. Sci., 2010; 2: 282 .; Moritz M, Chem. Eng J., 2013; 228: 596-613 .; Iravani S. ISRN., 2014; Especially, in medical field, it is applied to medical image, biosensor, gene target and drug delivery using nanoscale magnetism.

Among the various metals, silver has been used primarily in the treatment of bacterial infections as it exhibits excellent antibacterial properties. Silver nanoparticles in the form of nanoparticles are known to reduce cytotoxicity and increase the effect of antibacterial agents over their ionic forms, and thus they are applied to many fields for antibacterial, anti-inflammatory or infectious prevention. Gold has also been utilized in a variety of fields, including drug delivery, photo thermal therapies, immune chromatographic identification of pathogens in specimens, biosensors, and DNA markers, with its unique surface plasmon vibrations. In addition, metal nanoparticles exhibit excellent biocompatibility and are expected to be used as drug carriers because they are easily deformed and have high cell permeability (Narsireddy Amreddy et al., International Journal of Nanomedicine 2015: 10 6773-6788, Jean-Pascal Piret et al., Nanoscale, 2012, 4, 7168-7184).

Generally, chemical synthesis methods and physical production methods are used for the production of metal nanoparticles. The chemical synthesis method is relatively simple, but the cost is high and the toxicity of the reagent is problematic. In addition, physical methods have the disadvantage that it is difficult to control the size of nanoparticles, and it is difficult to produce effective metal nanoparticles because of the problems such as requiring expensive manufacturing facilities.

For this reason, researches on a method for producing environmentally friendly metal nanoparticles containing a natural extract as an active ingredient have been extensively studied. However, conventional methods for synthesizing metal nanoparticles by bacteria or strains require a long time for synthesis from days to days to several days. In order to stabilize the synthesized nanoparticles, a cap such as an additional reducing agent or a stabilizer There is a disadvantage that a capping agent is required. Therefore, development of metal nanoparticles overcoming the manufacturing time limit is still required as compared with a synthetic method which is still environmentally friendly, stable, chemical, or physical.

Under these circumstances, the inventors of the present invention have made extensive efforts to develop a method for producing metal nanoparticles more effectively. As a result, it has been confirmed that metal nanoparticles exhibiting antimicrobial and antioxidative activities can be prepared using black ginseng extract, Thus completing the present invention.

It is an object of the present invention to provide a composition for preparing metal nanoparticles, which comprises black ginseng extract as an active ingredient.

Another object of the present invention is to provide a method for producing metal nanoparticles, which comprises reacting the metal nanoparticle-forming composition with a metal precursor.

Still another object of the present invention is to provide the metal nanoparticles produced by the above-mentioned production method.

It is still another object of the present invention to provide an antimicrobial composition comprising the metal nanoparticles.

Yet another object of the present invention is to provide a pharmaceutical composition for antioxidation comprising the metal nanoparticles.

It is still another object of the present invention to provide an antioxidant health food composition comprising the metal nanoparticles.

It is still another object of the present invention to provide a cosmetic composition for antioxidation comprising the metal nanoparticles.

As one embodiment for achieving the above-mentioned object, the present invention provides a composition for the production of metal nanoparticles comprising black ginseng extract as an active ingredient.

The composition for preparing metal nanoparticles of the present invention can be used for preparing metal nanoparticles of uniform size without addition of a reducing agent or a stabilizer, which contains black ginseng extract as an active ingredient.

The term "black ginseng extract" of the present invention is not particularly limited, but may be prepared by the following method:

1) extracting black ginseng by adding an extraction solvent;

2) cooling the extract of step 1) and filtering; And

3) Concentrating the filtered extract of step 2) under reduced pressure and drying.

In the above method, the black gins of step 1) can be used without limitation, such as those prepared by processing white ginseng or commercially available ones. When the white ginseng is processed to produce black ginseng, black ginseng can be produced by repeating a process of sowing and drying white ginseng. The number of repetitions of the step of grinding and drying the white ginseng is not particularly limited. For example, 7 to 9 As another example, it is possible to repeat 9 times.

In the above method, the extraction solvent is not particularly limited as long as it can produce black ginseng extract, but water, C1 to C4 alcohols or a mixture thereof can be used. Specifically, ethanol, methanol or water can be used And more specifically 50% ethanol or water.

In the above method, conventional methods such as filtration, hot water extraction, immersion extraction, reflux cooling extraction and ultrasonic extraction can be used as the extraction method. The extraction solvent may be added in an amount of 0.1 to 10 times, preferably 0.3 to 5 times, to black ginseng, but is not limited thereto. In addition, the extraction temperature may be specifically 20 ° C to 150 ° C, but is not limited thereto, and the extraction time may be specifically 1 minute to 3 hours, but is not limited thereto.

In the above method, the vacuum concentration of the step 3) may be carried out by using a vacuum decompression concentrator or a vacuum rotary evaporator, but the present invention is not limited thereto. Specifically, the drying may be performed under reduced pressure, vacuum, boiling, spray drying, But is not limited thereto.

The term "metal nanoparticle" in the present invention means an ultra-fine particle having a size of 1 to 100 nm, which is made of metal, and exhibits a specific and diverse nature due to its small size. In the present invention, the metal nanoparticles may have different physiological activities depending on the production method. Specifically, the metal nanoparticles of the present invention prepared using the black ginseng extract include a physiologically active substance of the extract, It can have a much better biological activity than the prepared metal nanoparticles.

Specifically, the "metal" may be gold (Au) or silver (Ag), but is not limited thereto.

In the present invention, gold nanoparticles prepared using black ginseng extract may be used in combination as' gold nanoparticles' or 'Sg-AuNP (s)', and silver nanoparticles may be used as' silver nanoparticles' or 'Sg- (s) '.

In another embodiment, the present invention provides a method for producing metal nanoparticles, comprising the step of reacting the metal nanoparticle-forming composition with a metal precursor.

Since the method for producing metal nanoparticles according to the present invention uses the extract of black ginseng, which is a natural extract, it is more environmentally friendly than the conventional chemical synthesis method and physical synthesis method, and the metal nanoparticles It can be very usefully used in various industrial fields.

The term "metal precursor" of the present invention means a compound added to prepare metal nanoparticles. The metal precursor is not particularly limited as long as it can form metal nanoparticles by a reduction reaction. For example, the metal precursor may be a metal salt compound capable of forming metal nanoparticles by a reduction reaction.

For example, in the case of producing gold nanoparticles containing gold as a main component in metal nanoparticles, as an example of the metal precursor, tetrachloro gold (III) acid capable of forming gold nanoparticles by a reduction reaction (HAuCl ₄ ), NaAuCl ₃ , AuCl _3, and the like. As another example, tetrachloro gold (III) acid (HAuCl ₄ ) can be used.

In the case of producing silver nanoparticles containing silver as a main component in the metal nanoparticles, examples of the metal precursor include AgBF ₄ , AgCF ₃ SO ₃ and AgClO ₄ capable of forming silver nanoparticles by a reduction reaction ₄ , AgNO ₃ , AgPF ₆ , and Ag (CF ₃ COO). As another example, silver nitrate (AgNO ₃ ) can be used.

In the production method of the present invention, the concentration of the metal precursor may be 0.01 to 100 mM, specifically 0.01 to 50 mM, more specifically 0.01 to 10 mM, as long as it can produce metal nanoparticles.

In addition, in the production method of the present invention, the reaction temperature is not limited as long as the metal nanoparticles can be produced, but the reaction temperature is preferably 80 ° C or higher, more specifically 80 ° C or higher, 90 < 0 >C; When the metal is silver, the reaction temperature may be 60 占 폚 or higher, more specifically 60 to 90 占 폚.

In addition, in the production method of the present invention, as the reaction time increases, the level of the prepared metal nanoparticles increases, but the difference is dependent on the metal nanoparticles. As an example, in the case of gold nanoparticles, it may be from 1 minute to 1 hour, more specifically from 10 to 25 minutes, in the case of silver nanoparticles, specifically from 30 minutes to 7 hours, more specifically from 4.5 to 5 hours .

The method of the present invention may further include the step of recovering the metal nanoparticles by centrifuging the reaction product after the reaction is completed.

In another embodiment, the present invention provides the metal nanoparticles produced by the above-mentioned production method.

The metal nanoparticles of the present invention can be used for medical, biological analysis, fuel, and electronic component materials. Specifically, gold nanoparticles can be used as an organic solar cell, a sensor probe, a biological drug delivery system, a conductive material and a catalyst, due to inherent light conduction characteristics, and silver nanoparticles have optical properties, And antibacterial properties, it can be used as an antibacterial agent, a biosensor for quantitative detection, an optical analysis method, and the like. In addition, the metal nanoparticles of the present invention can control the optical and electronic characteristics of the particles by changing the size, shape, chemical characteristics of the surface, or aggregation state.

The shape of the metal nanoparticles produced by the method of the present invention may be generally spherical, but is not limited thereto.

The size of the crystals of the metal nanoparticles produced by the production method of the present invention may be 5 to 30 nm, more specifically 3 to 7 nm, more specifically 5 nm when the metal is gold; And may be from 5 to 10 nm, more specifically 8 nm, when the metal is silver.

In addition, it was confirmed that the metal nanoparticles were biologically stable because they were stably maintained even when they were left at pH conditions under different conditions at room temperature.

In another embodiment, the present invention provides an antimicrobial composition comprising the metal nanoparticles.

Here, the "metal nanoparticles" are as described above.

Since the metal nanoparticles of the present invention have an excellent antibacterial activity against pathogenic microorganisms, the composition containing the metal nanoparticles can be very usefully utilized in various industries.

The antimicrobial composition of the present invention may have an antimicrobial activity against bacteria, fungi or yeast, in particular Vibrio (Vibrio) genus, Salmonella (Salmonella), A stacking Pyrococcus (Staphylococcus), An Escherichia (Escherichia) in Bacillus (Bacillus) genus and Candida (Candida) may have an antimicrobial activity for at least one selected from the group consisting of a microorganism of the genus, more specifically, stearyl Pyrococcus (Staphylococcus) in microorganisms or Escherichia (Escherichia) in Microorganisms, more specifically Staphylococcus aureus and Escherichia coli coli ), but is not limited thereto.

In addition, the antimicrobial composition of the present invention is selected from the group consisting of lincomycin, oleandomycin, novobiocin, vancomycin, penicillin G, and rifampicin Lt; RTI ID = 0.0 > antibiotics < / RTI >

When the metal nanoparticles and the antibiotic are treated together, they exhibit an excellent antimicrobial activity against pathogenic microorganisms, rather than treating each of the metal nanoparticles or the antibiotic, and the composition containing the metal nanoparticles and the antibiotic can be very usefully utilized in various industries.

In another embodiment, the present invention provides a pharmaceutical composition for antioxidation comprising the metal nanoparticles.

Here, the "metal nanoparticles" are as described above.

The term "antioxidant" of the present invention refers to the removal of harmful free radicals, which means the protection of cells from oxidative damage by the free radicals.

Since the metal nanoparticles produced by the production method of the present invention exhibit antioxidative activity, antioxidant pharmaceutical compositions containing them as active ingredients can be used for the purpose of preventing, improving and treating diseases caused by active oxygen .

The diseases caused by the active oxygen include, but are not limited to, degenerative neurological diseases including arteriosclerosis, Lou Gehrig's disease, Parkinson's disease, Alzheimer's disease, proximal axillary sclerosis and Huntington's disease, myocardial infarction, angina pectoris, coronary artery disease , Cardiovascular diseases including ischemic heart diseases, ischemic brain diseases including stroke, diabetes, gastrointestinal diseases including gastritis and gastric cancer, aging, cancer, leukemia, aging, rheumatoid arthritis, hepatitis and atopic dermatitis And more specifically may be aging caused by active oxygen.

In another embodiment, the present invention provides a health food composition for antioxidation comprising the metal nanoparticles.

Here, the "metal nanoparticles" are as described above.

The gold nanoparticles produced by the production method of the present invention exhibit an antioxidative activity and can be prepared and taken in the form of foods that can prevent or improve diseases caused by active oxygen.

The term "food" of the present invention includes dairy products such as meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen and other noodles, gums, ice cream, various soups, drinks, tea, , A vitamin complex, a health functional food, and a health food, all of which include foods in a conventional sense.

The term "functional food" as used herein is the same term as "food for special health use" (FoSHU). In addition to nutritional supplementation, functional foods are processed so as to efficiently show the biological control function, It means food. Here, the term "function (surname)" means that the structure and function of the human body have a beneficial effect for health use such as controlling nutrients or physiological action. The food of the present invention can be prepared by a method commonly used in the art and can be prepared by adding raw materials and ingredients which are conventionally added in the art. In addition, the formulations of the food can also be produced without restrictions as long as they are formulations recognized as food. The composition for food of the present invention can be manufactured in various formulations, and unlike general pharmaceuticals, it has an advantage that there is no side effect that may occur when a food is used as a raw material for a long period of taking a medicine, The food of the invention can be ingested as an adjuvant to promote the effect of preventing or ameliorating the disease.

The health food refers to a food having an active health promotion effect or a health promotion effect compared to a general food, and a health supplement food refers to a health supplement food. In some cases, the terms health functional foods, health foods, and health supplements are used.

Specifically, the health functional food is a food prepared by adding metal nanoparticles to food materials such as beverage, tea, spice, gum and confectionery, or by encapsulation, powdering, suspension, etc., However, unlike general medicine, there is an advantage that there is no side effect that may occur when a food is used as a raw material for a long period of taking the medicine.

Since the food composition of the present invention can be routinely ingested, an excellent antioxidative effect can be expected, which is very useful.

The composition may further include a physiologically acceptable carrier. The carrier is not particularly limited and any carrier conventionally used in the art can be used.

In addition, the composition may contain additional ingredients which are commonly used in food compositions and which can improve odor, taste, vision and the like. For example, vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, panthotenic acid and the like. In addition, it may include minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu) It may also include amino acids such as lysine, tryptophan, cysteine, valine, and the like.

In addition, the composition can be used in combination with a preservative (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate), a disinfectant (such as bleaching powder and highly bleached white powder, sodium hypochlorite), an antioxidant (butylhydroxy anisole (BHA) (Sodium hypophosphate), bleach (sodium sulfite), seasoning (sodium MSG glutamate, etc.), sweeteners (hemicellulose, cyclamate, saccharin, A food additive such as a flavor (vanillin, lactones), a swelling agent (alum, D-tartrate, potassium hydrogen), an emulsifier, a thickening agent (glue), a covering agent, a gum base agent, a foam inhibitor, and food additives. The additives may be selected and used in appropriate amounts depending on the type of food.

The metal nanoparticles may be added directly or together with other food or food ingredients, and may be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to its intended use (prevention, health or therapeutic treatment). Generally, the food composition of the present invention may be added in an amount of not more than 50 parts by weight, specifically not more than 20 parts by weight, based on the food or beverage, when the food or drink is prepared. However, in case of long-term ingestion for health and hygiene purposes, the active ingredient may be contained in an amount not exceeding the above range and there is no problem in terms of safety.

As an example of the food composition of the present invention, it can be used as a health beverage composition. In this case, various flavors or natural carbohydrates can be added as an additional ingredient such as ordinary beverages. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose, sucrose; Polysaccharides such as dextrin, cyclodextrin; Xylitol, sorbitol, erythritol, and the like. Sweeteners include natural sweeteners such as tau Martin and stevia extract; Synthetic sweetening agents such as saccharin and aspartame, and the like can be used. The ratio of the natural carbohydrate may be generally about 0.01 to 0.04 g, specifically about 0.02 to 0.03 g per 100 mL of the composition of the present invention.

In addition to the above, the health beverage composition may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acid, protective colloid thickener, pH adjuster, stabilizer, Alcohols or carbonating agents, and the like. It may also contain flesh for the production of natural fruit juices, fruit juice drinks, or vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

In another embodiment, the present invention provides a functional cosmetic composition for antioxidation comprising metal nanoparticles.

Here, the "metal nanoparticles" are as described above.

The antioxidant cosmetic composition containing the metal nanoparticles can be used in the production of functional cosmetics capable of preventing or improving skin aging induced by active oxygen.

The term "cosmedical," or "cosmeceutical" refers to a cosmetic product that incorporates a special therapeutic function of a medicinal product. Unlike general cosmetics, it has a special function that emphasizes physiologically active effects and effects. Products that help to improve skin wrinkles, cosmetics prescribed by the Ordinance of the Ministry of Health and Welfare that help protect skin from burns or UV rays.

In the present invention, the functional cosmetic refers to a product that exhibits antioxidative activity in various functional cosmetics to help prevent aging of the skin. Specifically, the functional cosmetics may be a functional cosmetic for preventing skin aging comprising the gold nanoparticles as an effective ingredient , And the content of the gold nanoparticles contained in the functional skin cosmetic for preventing skin aging is also not particularly limited.

The functional cosmetic for preventing skin aging according to the present invention may further contain cosmetics conventionally used by containing the gold nanoparticles as an effective ingredient. For example, glycerol, propylene glycol, 1,3- Butylene glycol, sorbitol, polyethylene glycol, carboxyvinyl polymer, xanthan gum, carboxymethylcellulose, hydroxyethylcellulose, hydroxymethylcellulose, locust bean gum, allantoin, carrageenan and the like may be added; Paraffin wax, stearyl alcohol, carnauba wax, candelilla wax and calcium stearate, aluminum stearate, zinc stearate, and witchhazel can be used as the viscosity and hardness adjusting agent; As the ultraviolet absorber, butylmethoxydibenzoylmethane, octylmethoxycinnamate and the like can be used; Examples of the pigment include extender pigments such as titanium dioxide, fine particulate dioxides, kaolin, nylon powder, talc, sericite, mica and polymethyl methacrylate, and pigments such as yellow iron oxide, black iron oxide, red iron oxide, ultramarine, chromium oxide, Of a coloring pigment; Natural moisturizing agents such as 1,3-butylene glycol, concentrated glycerin, ethylene glycol, and chitin, chitosan, hypalonic acid, hyaluronic acid, lactic acid and glycolic acid can be used as the moisturizing agent; As the preservative, p-hydroxybenzoic acid esters, imidazolidinyl urea, and the like can be used. In addition, one or more of the above-mentioned components may be blended according to the characteristics of the product.

The functional cosmetic composition of the present invention may be prepared in any form conventionally produced in the art and may be in the form of a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, , Oil, powder foundation, emulsion foundation, wax foundation, and spray, but are not limited thereto. More specifically, it can be manufactured in the form of a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

When the formulation of the present invention is a paste, cream or gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters.

Using the method of the present invention, metal nanoparticles of uniform size can be prepared without adding any reducing agent or stabilizer in a short period of time, and the metal nanoparticles thus produced have antimicrobial activity and antioxidant activity Therefore, a composition containing the same can be usefully used in industry.

FIG. 1A is a graph showing the results of comparison of changes in the synthesis level of gold nanoparticles according to the synthesis temperature. FIG.
FIG. 1B is a graph showing the results of comparison of changes in the synthesis level of silver nanoparticles according to the synthesis temperature. FIG.
FIG. 1C is a graph showing the results of comparing changes in the synthesis level of gold nanoparticles according to the reaction time. FIG.
FIG. 1D is a graph showing the results of comparison of changes in the synthesis level of silver nanoparticles according to the synthesis time. FIG.
FIG. 1E is a photograph showing change in chromaticity of a synthetic reaction product according to synthesis reaction time of gold nanoparticles. FIG.
FIG. 1F is a photograph showing the chromaticity change of the synthesized reaction product according to the synthesis reaction time of the silver nanoparticles. FIG.
FIG. 2A is an electron micrograph showing the results of FE-TEM analysis of gold nanoparticles and silver nanoparticles, in which gold nanoparticles are shown on the left and silver nanoparticles are shown on the right.
FIG. 2B is a graph showing energy dispersive X-ray analysis (EDX) results of gold nanoparticles and silver nanoparticles. The upper part shows gold nanoparticles and the lower part shows silver nanoparticles.
FIG. 2C is a fluorescence microscope photograph showing the result of elemental mapping analysis of gold nanoparticles and silver nanoparticles, wherein gold is shown at the top and silver nanoparticles at the bottom.
FIG. 2D is a graph showing the XRD analysis results of the gold nanoparticles and the silver nanoparticles, wherein the top shows gold nanoparticles and the bottom shows silver nanoparticles.
FIG. 2E is a graph showing the results of particle size analyzer of gold nanoparticles through DLS. FIG.
FIG. 2f is a graph showing the results of particle size analyzer of silver nanoparticles through DLS. FIG.
FIG. 2G is a graph showing the results of FTIR analysis of gold nanoparticles and silver nanoparticles, wherein (A) represents silver nanoparticles, (B) represents gold nanoparticles, (C) represents a gold salt compound , (D) represents a silver salt compound, and (E) represents a black ginseng extract.
FIG. 3A is a graph showing the results of comparing antioxidative activities of gold and silver nanoparticles provided in the present invention. FIG.
FIG. 3B is a graph showing the results of comparing concentrations (IC ₅₀ ) required for removing 50% DPPH free radicals using the gold and silver nanoparticles provided in the present invention.
4A is a photograph showing antibacterial activity of silver nanoparticles of the present invention against E. coli .
4B is a photograph showing antimicrobial activity of silver nanoparticles of the present invention against S. aureus .
4C is a graph showing the results of analyzing the antibacterial activity of the silver nanoparticles of the present invention.
FIG. 5A is a graph showing the results of measurement of cell viability of human keratinocyte (HaCaT) and human breast cancer (MCF-7) cell lines treated with various concentrations of gold nanoparticles.
FIG. 5B is a graph showing the results of measuring cell viability of human keratinocyte (HaCaT) and human breast cancer (MCF-7) cell lines treated with various concentrations of silver nanoparticles.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example 1: Preparation of gold nanoparticles or silver nanoparticles using black ginseng extract

Example  1-1: Preparation of black ginseng extract

White ginseng was steamed at 100 ° C for 3 hours and then dried at 60 ° C for 24 hours. This operation was repeated 9 times to prepare black ginseng. The thus prepared black gum was pulverized to obtain a black gum powder, and 5 g of black gum powder was added with 100 ml of distilled water, followed by heating for 30 minutes for extraction. After the extraction was completed, the extract was filtered with a Wattman filter paper to obtain a liquid component. The supernatant obtained by centrifuging the obtained liquid component (10,000 rpm, 10 minutes) was used as a black ginseng extract.

Example  1-2: Preparation of gold nanoparticles

25 ml of distilled water was added to 5 ml of the black ginseng extract prepared in Example 1-1 to obtain a reaction solution. To the reaction solution, a gold salt compound (HAuCl ₄ -3H ₂ O) was added until a final concentration of 1 mM was reached, Particles were synthesized. The progress of the reaction was monitored by measuring the absorbance of the solution at constant time intervals. The color change of the solution in the reaction indicated the formation of gold nanoparticles. After the color change was confirmed, the reaction product was centrifuged (2,000 rpm, 10 minutes) to remove unnecessary components. Then, the synthesized nanoparticles were continuously washed with water, purified, and centrifuged (16,000 rpm, 15 minutes) to obtain gold nanoparticles in the form of a precipitate. Finally, the purified gold nanoparticles were air-dried overnight to produce gold nanoparticles in powder form.

Example  1-3: Preparation of silver nanoparticles

Silver nanoparticles in powder form were prepared using the method of Example 1-2 above, except that the silver salt compound (AgNO ₃ ) was used instead of the gold salt compound.

EXAMPLES 1-4: Effect of temperature on the production of nanoparticles

In order to analyze the effect of temperature on the nanoparticles of Examples 1-2 and 1-3, the synthesis reaction was carried out at various temperatures (23, 40, 60, 80 and 90 ° C) (Figs. 1A and 1B). The analysis was performed using a UV-Vis spectrometer. A small aliquot (200 μl) of the reacted reaction was dispensed into a UV-Vis spectrometer (2100Pro, Amersham, Biosciences Corp.) with a 10 mm path length correction cuvette. USA) in the range of 300-800 nm.

FIG. 1A is a graph showing the results of comparison of changes in the synthesis level of gold nanoparticles according to the synthesis temperature. FIG. As shown in FIG. 1A, the surface plasmon wavelength (λmax) of gold nanoparticles was confirmed at 548 nm. As the synthesis temperature increased, the maximum absorption peak of gold nanoparticles increased. As the synthesis temperature increased, Particles were synthesized in larger quantities. Particularly, it is found that the synthesis is preferably performed at a temperature of at least 80 ° C, since there is a large difference between the synthesis level of the gold nanoparticles synthesized at 60 ° C and the synthesis level of the gold nanoparticles synthesized at 80 ° C.

FIG. 1B is a graph showing the results of comparison of changes in the synthesis level of silver nanoparticles according to the synthesis temperature. FIG. As shown in FIG. 1B, the surface plasmon wavelength (λmax) of the silver nanoparticles was confirmed at 412 nm. It was also found that the silver nanoparticles were synthesized in larger amounts as the synthesis temperature also increased. In particular, it is preferable to synthesize the silver nanoparticles at a temperature of at least 60 ° C. because the silver nanoparticles synthesized at 40 ° C. exhibit a large difference in the synthesis level and the silver nanoparticles synthesized at 60 ° C.

Example  1-5: Effect of reaction time on nanoparticle preparation

In order to analyze the effect of the reaction time in the preparation of the nanoparticles of Examples 1-2 and 1-3, the synthesis reaction of the gold nanoparticles was carried out at 90 ° C. for 3, 10, 15, 20 and 25 minutes, The synthesis reaction of the nanoparticles was carried out at 90 DEG C for 3, 4 and 4.5 hours, and the synthesis level of the metal nanoparticles obtained therefrom was analyzed (FIGS. 1C and 1D). The analysis was carried out using a UV-Vis spectrometer.

FIG. 1C is a graph showing the results of comparing changes in the synthesis level of gold nanoparticles according to the reaction time. FIG. As shown in FIG. 1C, the surface plasmon wavelength (λmax) of the gold nanoparticles was confirmed at 548 nm. As the synthesis time increased, the maximum absorption peak of gold nanoparticles increased. As the synthesis time increased, Particles were synthesized in larger quantities. In particular, it is preferable to synthesize the gold nanoparticles synthesized for 3 minutes and the gold nanoparticles synthesized for 10 minutes at least for 10 minutes or longer.

FIG. 1D is a graph showing the results of comparison of changes in the synthesis level of silver nanoparticles according to the synthesis time. FIG. As shown in FIG. 1d, the surface plasmon wavelength (λmax) of the silver nanoparticles was confirmed at 412 nm. As the synthesis time of the silver nanoparticles increased, the silver nanoparticles were synthesized in larger amounts. In particular, it is preferable to synthesize silver nanoparticles for at least 4.5 hours, since the silver nanoparticles synthesized for 4 hours and the silver nanoparticles synthesized for 4.5 hours are significantly different from each other.

On the other hand, it was confirmed that the color of the synthetic reaction product changed with the lapse of time during the synthesis of the metal nanoparticles (FIGS. 1E and 1F).

FIG. 1E is a photograph showing the chromaticity change of the synthetic reaction product according to the synthesis reaction time of the gold nanoparticles. FIG. 1F is a photograph showing the chromaticity change of the synthesized reaction product according to the synthesis reaction time of the silver nanoparticles. As shown in FIGS. 1E and 1F, it was found that the appropriate reaction time of the metal nanoparticles can be confirmed by a method of confirming the chromaticity of the reactant without analyzing the surface plasmon wavelength of the metal nanoparticles.

Example 2: Structural characterization of gold nanoparticles or silver nanoparticles

Example  2-1: FE- TEM  analysis

The FE-TEM is used to determine the metal core size of the metal nanoparticles, while DLS is used to evaluate the hydrodynamic size (Z-average), including the capping layer of the nanoparticles. The sizes of the gold nanoparticles and silver nanoparticles were analyzed through the FE-TEM measurement (FIG. 2A).

FIG. 2A is an electron micrograph showing the results of FE-TEM analysis of gold nanoparticles and silver nanoparticles, in which gold nanoparticles are shown on the left and silver nanoparticles are shown on the right. As shown in FIG. 2A, it was confirmed that gold nanoparticles and silver nanoparticles were generally spherical with a varying size of 5 to 30 nm.

Example  2-2: Energy dispersive X-ray analysis EDX ) And element Mapping  analysis

EDX was prepared by placing one drop of the purified nanoparticles at 200 kV on a carbon coated copper grid and drying in an oven at 60 DEG C, using a JEM-2100F electron microscope (FIG. 2B).

FIG. 2B is a graph showing energy dispersive X-ray analysis (EDX) results of gold nanoparticles and silver nanoparticles. The upper part shows gold nanoparticles and the lower part shows silver nanoparticles. As shown in FIG. 2B, the existence of characteristic peaks of metallic gold and silver was confirmed at 2.2 keV and 3.3 keV, respectively

In addition, the purity of each metal nano-particle was confirmed through elemental mapping analysis of gold nanoparticles and silver nanoparticles, and the distribution of these nanoparticles was confirmed by combining the EDX results and the element mapping analysis results (FIG. 2C).

FIG. 2C is a fluorescence microscope photograph showing the result of elemental mapping analysis of gold nanoparticles and silver nanoparticles, wherein gold is shown at the top and silver nanoparticles at the bottom.

Example  2-3: XRD  analysis

XRD analysis was performed using an X-ray diffractometer, which operated at 40 kV, 40 mA, 1.54 A CuK alpha irradiation, a scan rate of 6 DEG / min and a step size of 0.02 DEG . The sample was scanned over 2? Over a range of 20-80 °. The average size of the metal nanoparticle crystals was calculated by the Debye-Scherrer equation:

D = 0.9? /? Cos?, D = 0.9? /? Cos?

Where D is the size of the microcrystalline per nm, λ is the wavelength of α irradiation per CuK nm, β is the full width at half maximum (FWHM) per radian, and θ is the median of the Bragg angle per radian. The XRD spectrum was recorded for identification of the crystallinity exhibited by the synthetic material (Fig. 2D).

FIG. 2D is a graph showing the XRD analysis results of the gold nanoparticles and the silver nanoparticles, wherein the top shows gold nanoparticles and the bottom shows silver nanoparticles. The characteristic peaks of gold nanoparticles (measured at 38.33 °, 44.31 °, 64.90 ° and 77.69 °) and silver nanoparticles (measured at 38.22 °, 44.24 °, 64.26 ° and 77.69 °) It was confirmed that the diffraction peaks of the lattice planes (111), (200), (220) and (311) interlocked with each other.

Example  2-4: Particle size analyzer

The particle size distribution of the nanoparticles suspended in distilled water was obtained by dynamic light scattering (DLS). The hydrodynamic (Z-average) diameter and PDI (polydispersity index) were evaluated at 25 占 폚. A dispersion medium of pure water having a refractive index of 1.3328, a viscosity of 0.8878 and a dielectric constant of 78.3 was used as a control (Figs. 2e and 2f).

FIG. 2E is a graph showing the results of particle size analyzer of gold nanoparticles through DLS. FIG. As shown in FIG. 2E, the Z-average size of the gold nanoparticles was 273.40 nm and the PDI value was 0.13, suggesting that the biosized gold nanoparticles are moderately polydisperse in nature.

FIG. 2f is a graph showing the results of particle size analyzer of silver nanoparticles through DLS. FIG. As shown in FIG. 2F, the Z-average size of the silver nanoparticles was 213.20 nm and the PDI value was 0.10, suggesting that the biosynthesized silver nanoparticles are monodisperse with a narrow distribution.

Example  2-5: FTIR (Fourier Transform- Infrared spectrometer ) analysis

FTIR measurements of gold nanoparticles and silver nanoparticles were performed using a PerkinElmer Spectrum One FTIR spectrometer at a resolution of 4000-450 cm ^-1 and 4 cm ^-1 (Figure 2g).

FIG. 2G is a graph showing the results of FTIR analysis of gold nanoparticles and silver nanoparticles, wherein (A) represents silver nanoparticles, (B) represents gold nanoparticles, (C) represents a gold salt compound , (D) represents a silver salt compound, and (E) represents a black ginseng extract.

As shown in FIG. 2g, the FTIR spectrum of the black ginseng root extract showed strong peaks at 3428 cm ^-1 , 1630 cm ^-1 and 1027 cm ^-1 , respectively, indicating the OH stretching vibration of the OH group in the polyphenylene flavonoid, the amino acid or protein residue , C = O stretching vibration associated with tertiary amines, and CO expansion of flavonoid esters. In addition, the shared median peak at 2927 cm ^-1 represents the CH stretching vibration of the methyl group.

On the other hand, the FTIR spectra of the silver nanoparticles and the gold nanoparticles showed peaks similar to those of the black ginseng root extract although their intensities were different. Specifically, the nano-particles exhibited a gold nanoparticle (3421, 2918, 1637 and 1021cm ^-1) weaker than the band around the biomolecule (3422, 2917, 1635 and 1025cm ^-1).

Through the FTIR spectral analysis, polyphenylene flavonoids, proteins and flavonoid esters in black ginseng root extracts were analyzed to be responsible for the reduction and capping of biosubstituted gold and silver nanoparticles.

Example  2-6: Stability Analysis

The stability of gold nanoparticles and silver nanoparticles was evaluated at the laboratory level.

Specifically, the stability of gold nanoparticles and silver nanoparticles was measured by using distilled water, 20 mM glycine-HCl buffer (pH 2.0), citric acid-sodium citrate buffer (pH 5.0), sodium phosphate buffer (pH 7.0), Tris- pH 8.0), 5% NaCl solution, 10% NaCl solution and 5% BSA solution. 900 쨉 l of each of the above solutions was added to 100 쨉 l of the metal nanoparticle solution, followed by reaction at 37 째 C. The gold and silver nanoparticles were found to be stable if there was no significant difference in surface plasmons using a UV-Vis spectrophotometer.

As a result, the surface plasmon wavelengths of DW, pH 2, pH 7, pH 8, 10% NaCl and 5% BSA solution of gold nanoparticles and silver nanoparticles showed the minimum wavelength shift (0-5 nm) It looked. In the case of gold nanoparticles, there was no shift of the surface plasmon wavelength in all the predetermined conditions even after one month. In the case of silver nanoparticles, long term stability was observed in DW and 5% BSA after one month. These results suggest that gold nanoparticles and silver nanoparticles are stable at physiological pH conditions.

Example  3: Functional characterization of gold nanoparticles or silver nanoparticles

Example  3-1: Antioxidant activity analysis of gold nanoparticles and silver nanoparticles

The antioxidant activity of gold nanoparticles (BGAuNPs), silver nanoparticles (BGgNPs), gold salt compounds and silver salt compounds was confirmed using the free radical scavenging activity of DPPH (1,1-diphenyl-2-picrylhydrazyl). To 160 μl of a 0.1 mM methanol solution of DPPH, 40 μl of a sample solution of various concentrations (2-20 μg / ml) was added. Galactic acid (GA) was used as a positive control. After 30 minutes, the absorbance at 517 nm was measured, and the free radical scavenging activity was calculated by applying the following equation, and the concentration was compared according to the concentration (FIG. 3A). At this time, gallic acid (GA) was used as a negative control group and black ginseng extract (BG) was used as a positive control group.

Free radical scavenging activity = [(control absorbance - sample absorbance) / control absorbance] × 100

At this time, sample absorbance was measured in DPPH solution added with nanoparticles, and control absorbance was measured in DPPH solution not added nanoparticles.

FIG. 3A is a graph showing the results of comparing antioxidative activities of gold and silver nanoparticles provided in the present invention. FIG. As shown in FIG. 3A, the silver nanoparticles provided by the present invention showed antioxidative activity similar to that of the black ginseng extract, and the gold nanoparticles provided by the present invention showed a somewhat lower antioxidative activity than the silver nanoparticles , A silver salt compound or a gold salt compound.

On the other hand, IC ₅₀ values indicating the concentration of the sample required to remove the 50% DPPH free radical of each sample showing the antioxidative activity were compared (Fig. 3B).

FIG. 3B is a graph showing the results of comparing concentrations (IC ₅₀ ) required for removing 50% DPPH free radicals using the gold and silver nanoparticles provided in the present invention. As shown in FIG. 3B, the IC ₅₀ value showed the highest level of the silver nanoparticles provided by the present invention and the gold nanoparticles were somewhat higher than those of the gallic acid and black ginseng extracts, as compared with the gallic acid and black ginseng extracts used as the control group. Respectively.

Example  3-2: Antimicrobial activity analysis of silver nanoparticles

The antimicrobial activity of silver nanoparticles against E. coli and S. aureus on an MHA (Mueller-Hinton agar) plate was analyzed using a known disk diffusion method. In this assay, 100 μl of the test strain cultured in LB medium at a density of 0.1 in MHA plate was plated and incubated overnight. As a control, a standard antimicrobial disk of neomycin was used. Next, fresh silver nanoparticle solutions were absorbed into sterile paper disks at various concentrations (15, 30 and 45 / / disc). The plate was incubated at 37 DEG C for 24 hours. After incubation was terminated, the inhibition regions around each disk were compared (Figs. 4A-4C).

FIG. 4A is a photograph showing the antibacterial activity of silver nanoparticles of the present invention against E. coli , FIG. 4B is a photograph showing antibacterial activity of silver nanoparticles of the present invention against S. aureus And FIG. 4C is a graph showing the results of analyzing the antibacterial activity of silver nanoparticles of the present invention.

As shown in FIGS. 4A to 4C, the silver nanoparticles provided in the present invention exhibit antimicrobial activity against Staphylococcus aureus belonging to Gram-positive bacteria as well as Escherichia coli belonging to Gram-negative bacteria, and the antibacterial activity is increased in a concentration- Respectively.

Example  3-3: Cytotoxicity analysis of gold nanoparticles and silver nanoparticles

Human keratinocyte (HaCaT) and human breast cancer (MCF-7) cell lines were obtained. The cell line was inoculated into a DMEM medium containing 10% FBS and 1% penicillin / streptomycin according to a known method and cultured at 37 ° C and 5% CO ₂ . The gold nanoparticles or silver nanoparticles of the present invention were added to each of the cultured cells at a concentration of 1, 10 and 100 μg / ml, cultured for another 72 hours, and then the viability of these cells was measured by MTT assay (Figs. 5A and 5B). At this time, as a control group, a cell line not treated with gold nanoparticles or silver nanoparticles was used.

FIG. 5A is a graph showing the results of measurement of cell viability of human keratinocyte (HaCaT) and human breast cancer (MCF-7) cell lines treated with various concentrations of gold nanoparticles. As shown in FIG. 5A, it was confirmed that the gold nanoparticles provided by the present invention had no effect on the cells even when treated in a large amount.

Therefore, it was found that the gold nanoparticles of the present invention did not show cytotoxicity at all.

FIG. 5B is a graph showing the results of measuring cell viability of human keratinocyte (HaCaT) and human breast cancer (MCF-7) cell lines treated with various concentrations of silver nanoparticles. As shown in FIG. 5B, it was confirmed that the silver nanoparticles provided by the present invention had an effect of reducing the survival rate of cells when treated in a large amount, unlike the gold nanoparticles.

Claims (19)

A composition for the preparation of metal nanoparticles, comprising a black ginseng extract as an active ingredient.
The method according to claim 1,
Wherein the metal nanoparticles are gold nanoparticles or silver nanoparticles.
The method according to claim 1,
Wherein the extract is obtained by extracting black ginseng with a solvent selected from the group consisting of water, C1 to C4 alcohols and mixed solvents thereof.
A method for producing metal nanoparticles, comprising: reacting the composition of claim 1 with a metal precursor to obtain a reaction product comprising metal nanoparticles.
5. The method of claim 4,
Wherein the metal precursor is a metal salt compound capable of forming metal nanoparticles by a reduction reaction.
6. The method of claim 5,
Wherein the metal salt compound is a gold salt compound or a silver salt compound.
The method according to claim 6,
Wherein the gold salt compound is a compound selected from the group consisting of tetrachloro gold (III) acid (HAuCl ₄ ), NaAuCl ₃ , AuCl _3, and combinations thereof.
The method according to claim 6,
Wherein the silver salt compound is a compound selected from the group consisting of AgBF ₄ , AgCF ₃ SO ₃ , AgClO ₄ , AgNO ₃ , AgPF ₆ , Ag (CF ₃ COO), and combinations thereof.
5. The method of claim 4,
Wherein the concentration of the metal precursor used in the reaction is 0.01 to 100 mM.
5. The method of claim 4,
Wherein the reaction is carried out at 10 to < RTI ID = 0.0 > 100 C. < / RTI >
5. The method of claim 4,
Wherein said step of reacting said composition with a gold salt compound for 1 minute to 1 hour to obtain a reaction product comprising gold nanoparticles.
5. The method of claim 4,
Wherein said step of reacting said composition with a silver salt compound for 30 minutes to 7 hours to obtain a reaction product comprising silver nanoparticles.
5. The method of claim 4,
Wherein the method further comprises centrifuging the reaction product to recover the metal nanoparticles.
A metal nanoparticle produced by the method of any one of claims 4 to 13.
A composition for antimicrobial use, comprising the metal nanoparticles of claim 14.
16. The method of claim 15,
Wherein the composition has an antimicrobial activity against microorganisms belonging to the genus Staphylococcus or Escherichia genus.
A pharmaceutical composition for antioxidation comprising the metal nanoparticles of claim 14.
A health food composition for antioxidation comprising the metal nanoparticles of claim 14.
A functional cosmetic composition for antioxidation comprising the metal nanoparticles of claim 14.

Images