KR20110059394A - Antibiotic cosmetic composition comprising alginate, fucoidan or silver-alginate having low molecular weight - Google Patents

Abstract

PURPOSE: An antibacterial cosmetic composition containing low molecular alginate, fucoidan, and silver-alginate is provided to safely ensure antibacterial activity and antioxidation effects. CONSTITUTION: An antibacterial cosmetic composition contains low molecular alginate having a molecular weight of 150 kDa or less; low molecular fucoidan having a molecular weight of 35 kDa or less; low molecular silver-alginate having a particle size of 0.1-3.4 um. The low molecular silver-alginate is prepared by adding 1.5-4.5 weight% of hydrogen peroxide to a solution containing 0.1-2.0 weight% of alginate; irradiating an electron beam to the mixture solution at 1.0-3.0MeV; adding a silver nitrate(AgNO_3) solution; and collecting precipitate. The cosmetic composition is manufactured in the form of skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, cleansing foam, cleansing lotion, body lotion, or body cleanser.

Description

Antibiotic cosmetic composition comprising alginate, fucoidan or silver-alginate having low molecular weight

The present invention relates to an antimicrobial cosmetic composition having excellent antibacterial activity and no cytotoxicity, and relates to an antimicrobial cosmetic composition containing a low molecular alginate, a low molecular fucoidan, or a low molecular silver-alginate as an active ingredient.

Alginate contained in seaweed such as seaweed and kelp has been widely used in various fields such as medical materials due to the discovery of antimicrobial and unique physiological functional effects, and the demand is greatly increased.

Alginate is a kind of seaweed polysaccharide and is composed of α-L-glulonate [G] and β-D-manruronate [M], and is a homopolymer block of MM block and GG block. Heteropolymer blocks such as homopolymer blocks or MG blocks are alternating polysaccharides composed of α-1,4 or β-1,4 bonds.

The alginate has a property of forming a gel when reacted with a metal ion. Such a gel has a property of adsorbing a metal ion. The reaction zone where alginate binds to metal ions is known because metal ions bond between carboxyl groups of uronic acid, a component of alginate, to form a three-dimensional network gel. The metal ions between the polyguluronic acid segment (G) of the alginate and the polymannuronic acid segment (M) form cross-links, forming a network structure, and uronic acid. It is reported that the array type of is related to the metal ion binding ability, and the lower the M / G ratio, the better the removal ability of heavy metals.

In addition, alginate can be used as a medicinal and bioactive substance such as antitumor agent, wound healing agent and wound dressing. In particular, alginate can be used as a hydrophilic gel to quickly absorb body fluids. When saline is added to the coating even when adhering to the wound, the insoluble calcium salt is converted into a soluble sodium alginate, which facilitates peeling. Third, the fibers remaining on the wound have biodegradation properties.

Therefore, in recent years, it has been applied chemically in various fields using these advantages of alginic acid, and researches are being conducted to prepare alginate as a gel to use as an enzyme fixative or to prepare a film formulation and a capsule. . In addition, the function of alginate as a dietary fiber is known, and it is also used as a dietary supplement by applying that alginate has a property of releasing harmful metals in vitro by combining with harmful metals.

Fucoidan is a major polysaccharide that forms brown algae with laminaran and alginate, and is contained in kelp and seaweed 3 to 5%. Fucoidan is a sulfated heteropolysaccharide, containing galactose, mannose, xylose, glucuronic acid, etc., mainly having L-fucose having a backbone composed of α-1,2 or α-1,3 bonds. In addition, fucoidan has been reported to have the pharmacological efficacy exhibited in polyanionic charges, for example anticoagulant, anti-cancer, anti-virus (anti- HIV (including HIV), anti-tumor, immune function improvement, antioxidant effects are known.

On the other hand, when oily foods and fast foods are settled as a general food culture in Korea, the number of patients with skin diseases including acne is increasing rapidly due to excessive sebum secretion and the activity of skin bacteria caused by the westernization of the diet. In particular, the domestic acne treatment market has grown by more than 50% from 11.3 billion won to 17.5 billion won over the past five years, and the trouble care cosmetics market, which is a treatment cosmetic for acne, is also gradually increasing. Therefore, research on the development of technology of functional cosmetics having antibacterial activity in accordance with this consumption trend is being activated.

Acne is caused by excessive sebum secretion, exfoliation of skin and inflammatory reactions caused by stimulation of free fatty acids produced by lipase of propionibacterium acne, an anaerobic dermal fungus, and various small molecule substances. The method for treating acne may include, for example, an anti-androgen agent that inhibits sebum production, a nonsteroidal anti-inflammatory agent that acts as an anti-inflammatory agent, an antibacterial agent such as lesochinol, sulfur, salicylic acid, benzoyl peroxide, and isotretinoin, and There is a method for inhibiting the activity of propionibacterium acne bacteria by treating antibiotics such as keratin release agent, tetracycline, erythromycin, and macrocycline on the skin.

In addition, skin diseases caused by dermatophytes include skin diseases such as athlete's foot and dandruff in addition to acne, and methods of treating these diseases are also used to inhibit the growth of dermatophytes by antibacterial and antifungal agents as described above. In particular, cosmetics using antibacterial and antibiotics and external skin preparations are mainly used. These methods are easy to cause skin irritation, may cause side effects such as phototoxicity, and irritant dermatitis with the possibility of causing resistant bacteria by long-term use. It is known that there are problems such as secondary bacterial infection. In addition, there is a surgical treatment such as scrub removal as a treatment method of acne, but the surgical treatment has a problem that the skin care should be continued after the procedure.

Therefore, in recent years, in order to prevent or suppress the development of skin diseases caused by acne and various bacteria, research into natural products with high antibacterial activity has been focused to develop new cosmetics or external preparations for skin without any side effects on the human body including skin. It is becoming. Looking at the related arts related to this, the Republic of Korea Patent Publication No. 2009-0105491 discloses the contents of the cosmetic composition containing a natural antibacterial complex consisting of lotus leaf, Jibuja, Jigolpi extract, and the Republic of Korea Patent Publication No. 2009-0084118 A functional cosmetic composition having an anti-inflammatory, antioxidant and whitening and antimicrobial effect, including a herbal extract, has been disclosed. The Republic of Korea Patent Publication No. 2009-0055261 discloses and improves and treats acne containing a joiner extract as an active ingredient. A description of the composition is disclosed.

   However, in the case of antibacterial cosmetics using the natural products developed so far, there is a problem in that the propagation of microorganisms and unpleasant odors occur because natural ingredients are more contaminated than synthetic ingredients, and thus a large amount of antiseptic or antibiotic According to the addition of the same antimicrobial chemicals there is a problem that the skin irritation occurs due to a number of preservatives.

Therefore, it is urgent to develop functional antibacterial cosmetics using new natural ingredients with excellent antibacterial activity without causing skin irritation.

Accordingly, an object of the present invention is to provide an antimicrobial cosmetic composition comprising a low molecular alginate, a low molecular fucoidan or a low molecular silver-alginate as an active ingredient in order to provide a cosmetic composition having no skin irritation and excellent antimicrobial activity.

In order to achieve the above object of the present invention, the present invention is one selected from the group consisting of a low molecular weight alginate having a molecular weight of 150,000 Da or less, a low molecular weight fucoidan having a molecular weight of 35,000 Da or less and a low molecular weight silver-alginate having a particle size of 0.1 to 3.4 μm It provides an antimicrobial cosmetic composition comprising the above in an amount of 0.01 to 1.0% (w / v), respectively, based on the total weight of the composition.

In one embodiment of the present invention, the low molecular weight alginate or low molecular fucoidan is added to the aqueous solution containing 0.1 to 2.0% by weight of alginate or fucoidan in the amount of 1.5% to 4.5% by weight of hydrogen peroxide, the hydrogen peroxide is added It can be prepared by the step of irradiating an electron beam of 1.0 ~ 3.0 MeV to the mixed solution.

In one embodiment of the present invention, the low molecular weight silver-alginate is added to the aqueous solution containing 0.1 to 2.0% by weight of alginate in an amount of 1.5% to 4.5% by weight of hydrogen peroxide, the hydrogen peroxide is added to the mixed solution It can be prepared by irradiating an electron beam of 1.0 ~ 3.0 MeV, reacted by adding a silver nitrate (AgNO ₃ ) solution, and then recovering the precipitate.

In one embodiment of the present invention, the antimicrobial cosmetic composition may have an antimicrobial activity and radical scavenging activity against a strain selected from the group consisting of Escherichia coli, Staphylococcus aureus, acne and anthrax.

In one embodiment of the present invention, the minimum growth inhibition concentration (MIC) for the strain of the low molecular weight alginate, low molecular fucoidan and low molecular weight silver alginate may be 400 ~ 10,000ppm, 5,000 ~ 10,000ppm and 300 ~ 400ppm, respectively.

In one embodiment of the present invention, the composition is a skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutrition cream, moisturizing cream, hand cream, foundation, essence, It may be one of the formulations selected from the group consisting of nutrient essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser.

Since the antimicrobial cosmetic composition according to the present invention contains low-molecular alginate, fucoidan or silver-alginate prepared by a low molecular weight technology using hydrogen peroxide and electron beam irradiation, it has excellent antibacterial activity against harmful skin strains that reside deep in the skin and pores. In addition, since it is excellent in antioxidant activity to remove radicals and does not have toxicity to cells, and has excellent stability to skin, it can be usefully used in the manufacture of functional cosmetics for improving skin troubles such as acne and atopy. .

The present invention relates to an antimicrobial cosmetic composition having excellent antimicrobial activity, and more specifically, among a group consisting of a low molecular weight alginate having a molecular weight of 150,000 Da or less, a low molecular weight fucoidan having a molecular weight of 35,000 Da or less and a low molecular weight silver-alginate having a particle size of 0.1 to 3.4 μm. It is characterized by providing an antimicrobial cosmetic composition comprising at least one selected from 0.01 to 1.0% (w / v) based on the total weight of the composition.

In the present invention, low molecular weight alginate, fucoidan or silver-alginate was used to prepare a cosmetic composition having excellent antimicrobial activity.Alginate and fucoidan are components contained in seaweeds such as seaweed and seaweed, in particular, algae extracts protect skin. It is used as a raw material of cosmetics for skin because it not only hydrates and softens, but also has no toxicity and no irritation to skin and eyes.

However, alginate and fucoidan, which are polysaccharides contained in seaweed, belong to a relatively large particle size, and when they are made into nanoparticles with low molecular weight, they may increase the absorption capacity to skin pores (size; about 20-50 μm). In view of the fact that the antimicrobial effect of these ingredients can be enhanced, the present invention used low-differentiated alginate or fucoidan in the preparation of the cosmetic composition, and further added silver (Ag) to the low molecular alginate. The use of the low molecular weight silver-alginate prepared in the preparation of the cosmetic composition provides a very good antimicrobial cosmetic composition to which the antimicrobial properties of silver are further added.

On the other hand, alginate generally represented by the following structural formula has a large molecular weight, high viscosity, poor solvent properties, there are a lot of difficulties in using for various uses, such as medical, cosmetics and food. In addition, there is a difficulty in workability because a technique for controlling the molecular weight according to the purpose and mass production with high efficiency is not established.

                             <Structure of Alginate>

In addition, alginates that can be used to prepare the low molecular weight alginates according to the invention include, but are not limited to, esterified forms of alginates, as well as monovalent salts of alginates, such as sodium and potassium alginates. For example, propylene glycol alginate can be used. Further examples of alginates that may be used in the present invention are magnesium alginate and 3-ethanol amine alginate.

Therefore, the low molecular weight alginate that may be used in the cosmetic composition according to the present invention may use a low molecular weight alginate having a molecular weight of 150,000 Da or less, which is low molecular weight by electron beam irradiation, and preferably 1.0 to an aqueous solution containing 0.1 to 2.0% by weight of alginate. A low molecular weight alginate having a molecular weight of 150,000 Da or less prepared by irradiating an electron beam of 3.0 MeV may be used.

In addition, fucoidan is a saccharide in the form of a viscous polymer having a weight average molecular weight of 30,000-2,000,000 Da, which is generally represented by the following structural formula, and is mainly included in brown algae such as seaweed and kelp, and especially fucoidan extracted from seaweed spores. It is known that the average molecular weight is about 170,000 Da. In addition, the functions of brown algae-derived fucoidan have been reported to have activities such as immunopotentiation, anti-tumor action, suppression of blood glucose elevation and blood coagulation, triglyceride and cholesterol lowering action (Scand. J. Urol. Nephrol). ., 40, 1, 2006; Journal of the Korean Chemical Society, 46, 2, 2002).

                            <Structure of Fucoidan>

On the other hand, the low molecular fucoidan according to the present invention can use a low molecular fucoidan having a molecular weight of 35,000 Da or less, preferably prepared by irradiating an electron beam of 1.0 ~ 3.0 MeV in an aqueous solution containing 0.5 to 2.0% by weight of fucoidan A low molecular weight fucoidan having a molecular weight of 35,000 Da or less can be used.

In addition, the low molecular weight silver-alginate according to the present invention may have a molecular weight of 15,000 Da or less and a low molecular weight silver-alginate having a particle size of 0.1 to 3.4 μm with a content of silver (Ag) of 35% or more. In addition, the low molecular weight silver-alginate is added to the aqueous solution containing 0.1 to 2.0% by weight of alginate in an amount of 1.5% to 4.5% by weight of hydrogen peroxide, 1.0-3.0 MeV electron beam is added to the mixed solution to the hydrogen peroxide The low molecular weight silver-alginate prepared by the step of reacting the low molecular weight alginate with a molecular weight of 5,000 Da or less by adding a silver nitrate (AgNO ₃ ) solution and then recovering the precipitate may be used, and the low molecular weight silver alginate used in the present invention may be used. The structural model of is as shown below.

                       Structural Formula of Low Molecular Silver-Alginate

As described above, the low molecular weight silver-alginate according to the present invention may be prepared according to the method described in the patent application No. 2008-0065856, which the inventors have previously applied for, and the patent application No. The entirety of 2008-0065856 is hereby incorporated by reference.

On the other hand, the low-molecular alginate, low-molecular fucoidan and low-molecular silver-alginate contained in the cosmetic composition of the present invention is characterized by having excellent antimicrobial activity against harmful strains causing skin diseases, and as harmful strains causing the skin diseases. May be, but are not limited to, E. coli, Staphylococcus, Acne cocci and Anthrax.

According to one embodiment of the invention, the antimicrobial activity and minimal growth of E. coli, staphylococcus, acne and anthrax strains for low molecular alginate, low molecular fucoidan and low molecular silver alginate used in the preparation of the cosmetic composition of the present invention Investigation of the inhibitory concentration (MIC) showed that low molecular alginate, low molecular fucoidan, and low molecular silver-alginate all inhibited the growth of these strains.Especially, the low molecular alginate of Escherichia coli and Staphylococcus at low concentrations of 400 ppm It was shown that there is an activity to inhibit (see Table 6), in the case of low-molecular fucoidan was shown to have an inhibitory activity against Staphylococcus aureus at 5000ppm and to E. coli and acne bacteria at 10000ppm (Figs. 1 to 3). Low concentration of 350 ppm for low molecular weight silver-alginates Standing in E. coli, it was found to be active to inhibit both Staphylococcus and acne bacteria (see Table 13 and Figure 4).

Accordingly, the present inventors have found that despite the use of low concentrations of low-molecular alginate, low-molecular fucoidan, and low-molecular silver-alginate, the present inventors have found that there is an activity that inhibits the growth of harmful skin strains. Was found to be very good.

Therefore, the minimum growth inhibition concentration (MIC) of the low molecular alginate, low molecular fucoidan and low molecular silver-alginate according to the present invention is 400 to 10,000 ppm of low molecular alginate, 5000 to 10000 ppm of low molecular fucoidan, and 300 to 400 ppm of low molecular silver alginate. Can be.

  The minimum growth inhibitory concentration (MIC: Minimum Inhibitory Concentration) refers to a minimum concentration capable of inhibiting or inhibiting the growth of bacteria or bacteria. The lower the concentration, the better the inhibitory activity of the bacteria. have.

In the present invention, the term "antimicrobial" includes not only inhibiting the growth of pathogenic microorganisms such as bacteria, fungi, etc., but also inhibiting their survival and sterilizing.

In addition, according to another embodiment of the present invention, after treating the low-molecular alginate, low-molecular fucoidan and low-molecular silver-alginate at the minimum growth inhibition concentration, the antimicrobial activity was measured by measuring the cell concentration of harmful bacteria over time. As the treatment time increases, the growth rate of each strain is gradually decreased (see Table 7 to Table 12).

Furthermore, the low molecular weight alginate, low molecular weight fucoidan and low molecular weight silver-alginate contained in the cosmetic composition of the present invention are characterized by having antioxidant activity to eliminate radicals and at the same time, no toxicity to cells.

Radicals generated inside cells cause various problems such as wrinkle formation, skin cancer, cell death, rheumatoid arthritis, atopic dermatitis and acne through the action of destroying the cells or cutting or cross-linking the connective tissue of the skin dermis. It is known. Therefore, the radical removing effect can improve and treat various skin diseases resulting from harmful bacteria.

In this regard, according to one embodiment of the present invention, it was analyzed whether there is a radical scavenging activity for the low molecular alginate, low molecular fucoidan and low molecular silver alginate contained in the antimicrobial cosmetic composition of the present invention, as a result, low molecular alginate, low molecular Both fucoidan and low molecular weight silver-alginate were found to have antioxidant activity to remove radicals (see Tables 14 and 15), and also showed an increase in radical scavenging effect in proportion to the amount of treatment of each component (Figure 6 and 7, in particular low molecular weight silver-alginate, showed very good scavenging activity of radicals (see FIG. 8).

In addition, according to another embodiment of the present invention, the low molecular weight alginate, low molecular fucoidan and low molecular weight silver alginate contained in the antimicrobial cosmetic composition of the present invention was investigated as a result, it was found that there is no toxicity to cells (See Table 16).

Therefore, through the above results, the present inventors can use the low-molecular alginate, low-molecular fucoidan, or low-molecular silver-alginate in the manufacturing process of cosmetics to prepare antimicrobial cosmetics having excellent safety as well as antimicrobial activity and safety. I could tell the truth.

Therefore, the present invention can provide an antimicrobial cosmetic composition comprising at least one selected from the group consisting of a low molecular weight alginate having a molecular weight of 150,000 or less, a low molecular weight fucoidan having a molecular weight of 35,000 or less, and a low molecular weight silver-alginate having a particle size in the range of 0.1 to 3.4 μm.

The composition according to the present invention may include the low molecular weight alginate, low molecular fucoidan or low molecular weight silver alginate in the range of 0.01 to 1.0% (w / v) based on the total weight of the composition, wherein each component is 0.01 to the total weight of the composition If it is included in less than% (w / v) there is a problem that the antimicrobial activity is reduced, when included in excess of 1.0% (w / v) because only the amount of addition is large, there is no significant difference in antimicrobial activity.

In addition, the cosmetic composition according to the present invention may contain an adjuvant commonly used in the cosmetic field within the range not impairing the main effect of the present invention in addition to the above essential components. Such as hydrophilic or hydrophobic gelling agents, hydrophilic or hydrophobic active agents, preservatives, antioxidants, solvents, fragrances, fillers, blockers, pigments, odorants and dyes. The amount of these various adjuvants may be added in amounts conventionally used in the art, and in any case the adjuvants and their proportions may be selected so as not to adversely affect the desirable properties of the cosmetic composition according to the present invention.

In addition, the cosmetic composition according to the present invention may be prepared in any formulation prepared in the art. It can preferably be prepared in a formulation for skin application, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations , Wax foundations, sprays, and the like, but is not limited thereto. More preferably, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nourishing lotion, massage cream, nourishing cream, moisturizing cream, hand cream, foundation, essence, nutrition essence, pack, soap, cleansing It may be prepared in the form of a foam, cleansing lotion, cleansing cream, body lotion and body cleanser.

When the formulation of the composition according to the present invention is a paste, cream or gel, the carrier component is animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide. Lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component when the formulation of the composition according to the invention is a powder or a spray, and in the case of a spray, chloro Propellants such as fluorohydrocarbons, propane / butane or dimethylether may additionally be included. In addition, when the formulation of the composition according to the invention is a suspension, as a carrier component, liquid diluents such as water, ethanol or propylene glycol, ethoxylated isosteroyl alcohols, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters Suspending agents, microcrystalline cellulose, aluminum metahydroxide, bentonide, agar or tracant and the like can be used and when the formulation of the composition according to the invention is a surfactant-containing cleansing, an aliphatic alcohol sulfate as a carrier component. , Fatty alcohol ether sulfates, sulfosuccinic acid monoesters, isethionates, imidazolinium derivatives, fatty acid amide ether sulfates, alkylamidobetaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxyls Digested Glycerol Fatty Acid S Include Le can be used.

In one embodiment of the present invention, the low-molecular alginate, low-molecular fucoidan or low-molecular silver-alginate alone or mixed according to the present invention to prepare an essence containing the components, the antimicrobial activity of the prepared essence cosmetics of the present invention is analyzed As a result, the essence containing low molecular alginate, low molecular fucoidan or low molecular silver-alginate showed superior antimicrobial activity to the essence which did not contain the above components, and especially low molecular alginate, low molecular fucoidan and low molecular silver alginate In the case of the contained essence, the antimicrobial activity was found to be very good (see FIG. 9).

Therefore, the present invention can provide a method for producing an antibacterial cosmetic using the antimicrobial cosmetic composition according to the present invention, it can provide an antibacterial cosmetic prepared by the above method.

Hereinafter, the present invention will be described in detail by way of examples. However, these examples are intended to illustrate the present invention in more detail, and the scope of the present invention is not limited to these examples.

&Lt; Example 1 >

Preparation and Characterization of High / Low Molecular Alginate and Fucoidan

The present inventors first prepared a low molecular alginate and fucoidan to prepare an antimicrobial cosmetic composition using silver-alginate and fucoidan, and then measured the molecular weight and viscosity of the prepared low molecular alginate and fucoidan. First, the preparation of low molecular weight alginate uses electron beam irradiation (1.0 MeV or 2.5 MeV) and 30% hydrogen peroxide (H ₂ O ₂ , domestic) in an aqueous solution of alginate (sodium alginate, Junsei) at 2% (w / v). Low molecular alginate was prepared, and fucoidan (fucoidan, Haewon Biotech Co., Ltd.) was also prepared low molecular weight fucoidan in the same manner. Then, the molecular weight and viscosity of the low molecular weight alginate and fucoidan prepared by the above method were measured according to the content of hydrogen peroxide and the intensity of the electron beam used in the preparation, and Table 1 and Table 2 were prepared by electron beam irradiation of 1.0 MeV or 2.5 MeV. Molecular weight and viscosity results for low molecular alginate are described, and Tables 3 and 4 below describe molecular weight and viscosity results for low molecular fucoidan prepared by electron beam irradiation of 1.0 MeV or 2.5 MeV.

TABLE 1

Molecular Weight and Viscosity of Low Molecular Alginate Prepared by Electron Beam Irradiation (2.5 MeV)

Manufacturing example Hydrogen peroxide content% Electron beam intensity (kGy) Mw Mn PDI
(Mw / Mn) η _inh pH  Native alginate - 446,467 226,167 1.974 6.44 7.63 One A-2.5-HP0-2.5 0 2.5 92,187 46,768 1.971 1.88 6.13 2 A-2.5-HP0-5.0 0 5.0 51,786 26,310 1.968 1.36 6.13 3 A-2.5-HP0-10 0 10.0 30,809 15,688 1.964 1.09 5.83 4 A-2.5-HP0-15 0 15.0 20,709 10,573 1.959 0.96 4.84 5 A-2.5-HP0-20 0 20.0 13,717 7,033 1.950 0.87 5.32 6 A-2.5-HP1.5-2.5 1.5 2.5 55,671 28,277 1.969 1.41 5.53 7 A-2.5-HP1.5-5 1.5 5.0 23,817 12,147 1.961 1.00 5.19 8 A-2.5-HP1.5-10 1.5 10.0 9,055 4,672 1.938 0.81 5.01 9 A-2.5-HP1.5-15 1.5 15.0 5,948 3,098 1.920 0.77 4.82 10 A-2.5-HP1.5-20 1.5 20.0 3,617 1,918 1.886 0.74 4.67 11 A-2.5-HP3.0-2.5 3.0 2.5 68,102 34,572 1.970 1.57 5.39 12 A-2.5-HP3.0-5 3.0 5.0 33,917 17,261 1.965 1.13 5.15 13 A-2.5-HP3.0-10 3.0 10.0 9,832 5,065 1.941 0.82 4.94 14 A-2.5-HP3.0-15 3.0 15.0 7,501 3,885 1.931 0.79 4.83 15 A-2.5-HP3.0-20 3.0 20.0 4,394 2,312 1.901 0.75 4.80 16 A-2.5-HP4.5-2.5 4.5 2.5 80,533 40,867 1.971 1.73 5.57 17 A-2.5-HP4.5-5 4.5 5.0 44,794 22,769 1.967 1.27 5.40 18 A-2.5-HP4.5-10 4.5 10.0 30,032 15,294 1.964 1.08 5.16 19 A-2.5-HP4.5-15 4.5 15.0 8,278 4,279 1.935 0.80 4.82 20 A-2.5-HP4.5-20 4.5 20.0 5,171 2,705 1.912 0.76 4.94

TABLE 2

Molecular Weight and Viscosity of Low Molecular Alginate Prepared by Electron Beam Irradiation (1.0 MeV)

Manufacturing example Hydrogen peroxide content% Electron beam intensity (kGy) Mw Mn PDI
(Mw / Mn) η _inh 21 A-1-HP0-2.5 0 2.5 148,126 75,094 1.973 2.60 22 A-1-HP0-5.0 0 5.0 93,741 47,555 1.971 1.90 23 A-1-HP0-10 0 10.0 122,487 62,111 1.972 2.27 24 A-1-HP0-15 0 15.0 89,079 45,194 1.971 1.84 25 A-1-HP0-20 0 20.0 58,002 29,457 1.969 1.44 26 A-1-HP1.5-2.5 1.5 2.5 81,310 41,260 1.971 1.74 27 A-1-HP1.5-5 1.5 5.0 84,418 42,834 1.971 1.78 28 A-1-HP1.5-10 1.5 10.0 71,987 36,539 1.970 1.62 29 A-1-HP1.5-15 1.5 15.0 40,133 20,409 1.966 1.21 30 A-1-HP1.5-20 1.5 20.0 40,909 20,802 1.967 1.22 31 A-1-HP3.0-2.5 3.0 2.5 99,956 50,702 1.971 1.98 32 A-1-HP3.0-5 3.0 5.0 52,563 26,704 1.968 1.37 33 A-1-HP3.0-10 3.0 10.0 57,225 29,064 1.969 1.43 34 A-1-HP3.0-15 3.0 15.0 65,771 33,392 1.970 1.54 35 A-1-HP3.0-20 3.0 20.0 55,671 28,277 1.969 1.41 36 A-1-HP4.5-2.5 4.5 2.5 143,464 72,734 1.972 2.54 37 A-1-HP4.5-5 4.5 5.0 48,679 24,736 1.968 1.32 38 A-1-HP4.5-10 4.5 10.0 68,102 34,572 1.970 1.57 39 A-1-HP4.5-15 4.5 15.0 64,994 32,998 1.970 1.53 40 A-1-HP4.5-20 4.5 20.0 35,471 18,048 1.965 1.15

[Table 3]

Molecular Weight and Viscosity of Low Molecular Fucoidan Prepared by Electron Beam Irradiation (2.5 MeV)

Manufacturing example Hydrogen peroxide content% Electron beam intensity (kGy) Mn Mw PDI
(Mw / Mn) η _inh Native fucoidan - 0 50,702 99,956 1.971 1.98 41 F-1-HP0-2.5 0 2.5 33,392 65,771 1.970 1.54 42 F-1-HP0-5.0 0 5.0 25,523 50,233 1.968 1.34 43 F-1-HP0-10 0 10.0 19,622 38,579 1.966 1.19 44 F-1-HP0-15 0 15.0 15,688 30,809 1.964 1.09 45 F-1-HP0-20 0 20.0 12,147 23,817 1.961 1.00 46 F-1-HP1.5-2.5 1.5 2.5 32,211 63,440 1.970 1.51 47 F-1-HP1.5-5 1.5 5.0 22,376 44,017 1.967 1.26 48 F-1-HP1.5-10 1.5 10.0 17,655 34,694 1.965 1.14 49 F-1-HP1.5-15 1.5 15.0 11,754 23,040 1.960 0.99 50 F-1-HP1.5-20 1.5 20.0 9,000 17,602 1.956 0.92 51 F-1-HP3.0-2.5 3.0 2.5 30,244 59,556 1.969 1.46 52 F-1-HP3.0-5 3.0 5.0 26,310 51,786 1.968 1.36 53 F-1-HP3.0-10 3.0 10.0 19,229 37,802 1.966 1.18 54 F-1-HP3.0-15 3.0 15.0 13,327 26,148 1.962 1.03 55 F-1-HP3.0-20 3.0 20.0 12,147 23,817 1.961 1.00 56 F-1-HP4.5-2.5 4.5 2.5 3,885 7,501 1.931 0.79 57 F-1-HP4.5-5 4.5 5.0 23,950 47,125 1.968 1.30 58 F-1-HP4.5-10 4.5 10.0 7,819 15,271 1.953 0.89 59 F-1-HP4.5-15 4.5 15.0 18,835 37,025 1.966 1.17 60 F-1-HP4.5-20 4.5 20.0 15,294 30,032 1.964 1.08

[Table 4]

Molecular Weight and Viscosity of Low Molecular Fucoidan Prepared by Electron Beam Irradiation (1.0 MeV)

Manufacturing example Hydrogen peroxide content% Electron beam intensity (kGy) Mn Mw PDI
(Mw / Mn) η _inh Native fucoidan - 0 50,702 99,956 1.971 1.98 61 F-2.5-HP0-2.5 0 2.5 46,768 92,187 1.971 1.88 62 F-2.5-HP0-5.0 0 5.0 26,310 51,786 1.968 1.36 63 F-2.5-HP0-10 0 10.0 15,688 30,809 1.964 1.09 64 F-2.5-HP0-15 0 15.0 10,573 20,709 1.959 0.96 65 F-2.5-HP0-20 0 20.0 7,033 13,717 1.950 0.87 66 F-2.5-HP1.5-2.5 1.5 2.5 28,277 55,671 1.969 1.41 67 F-2.5-HP1.5-5 1.5 5.0 12,147 23,817 1.961 1.00 68 F-2.5-HP1.5-10 1.5 10.0 4,672 9,055 1.938 0.81 69 F-2.5-HP1.5-15 1.5 15.0 3,098 5,948 1.920 0.77 70 F-2.5-HP1.5-20 1.5 20.0 1,918 3,617 1.886 0.74 71 F-2.5-HP3.0-2.5 3.0 2.5 34,572 68,102 1.970 1.57 72 F-2.5-HP3.0-5 3.0 5.0 17,261 33,917 1.965 1.13 73 F-2.5-HP3.0-10 3.0 10.0 5,065 9,832 1.941 0.82 74 F-2.5-HP3.0-15 3.0 15.0 3,885 7,501 1.931 0.79 75 F-2.5-HP3.0-20 3.0 20.0 2,312 4,394 1.901 0.75 76 F-2.5-HP4.5-2.5 4.5 2.5 40,867 80,533 1.971 1.73 77 F-2.5-HP4.5-5 4.5 5.0 22,769 44,794 1.967 1.27 78 F-2.5-HP4.5-10 4.5 10.0 15,294 30,032 1.964 1.08 79 F-2.5-HP4.5-15 4.5 15.0 4,279 8,278 1.935 0.80 80 F-2.5-HP4.5-20 4.5 20.0 2,705 5,171 1.912 0.76

<Example 2>

Preparation of Silver Alginate

Silver-alginate to be used in the preparation of the antimicrobial cosmetic composition according to the present invention was prepared with silver-alginate using the low atomizing alginate prepared in Example 1 together with silver, and the preparation method has been previously filed by the present inventors. It was prepared according to the method described in Patent Application No. 2008-0065856, that is, for preparing low molecular weight alginic acid, the well-dried alginate was dissolved in hydrogen peroxide and then reacted at 50 ° C. for 3 hours in an ultrasonic atmosphere, and then the prepared silver nitrate solution was prepared. (silver nitrate / distilled water) was slowly dropped using a dropping funnel, and then reacted at room temperature for 1 hour, and the resultant was dropped in ethanol, washed with distilled water and ethanol, and dried under reduced pressure. Silver-alginate was prepared, and the low molecular weight alginate used was 5000 Da. The molecular weight of the sample was used. Further, in the preparation of the cosmetic composition and the antimicrobial activity test, the silver-alginate prepared contained 35% or more of silver.

<Example 3>

Antimicrobial test

<3-1> Antimicrobial Activity of Low Molecular Weight Alginate and Fucoidan

In order to examine whether each of the low molecular weight alginate and low molecular weight fucoidan prepared in Example 1 has antimicrobial activity, E. coli , Staphylococcus aureus subsp. Propionibacterium acnes) were each tested for antimicrobial activity . Culture conditions and methods for the respective strains are as described in Table 5 below.

TABLE 5

Culture Conditions and Methods of Strains

Name Culture condition Liquid badge Incubator Escherichia coli 37 ° C, aerobic culture ¹⁾ LB medium ³⁾ Aerobic chamber Staphylococcus 37 ° C, aerobic culture ¹⁾ BHI ⁴⁾ Aerobic chamber Acne 37 ° C., Anaerobic Culture ²⁾ TSB ⁵⁾ Anaerobic chamber Anthrax 37 ° C., Anaerobic Culture ²⁾ LB / TSB Anaerobic chamber

In Table 5 above For aerobic culture, put 100 ml of liquid medium in a 200 ml flask, sterilize at 121 ° C for 15 minutes using a high temperature and high pressure sterilizer, and then leave it on a clean bench until it reaches room temperature. After diluting the cultured bacteria (Cell Optical Density 2.0 or higher 2.0 × 10 ^{3 ~ 4} CFU / mL) by 50 times, inoculating 100 μl into the liquid medium and subcultured every 4 weeks to continuously maintain the activity of the bacteria. gave. The antimicrobial test was carried out by placing 4 ml of liquid medium in a 15 ml sterile test tube and properly diluting (diluted stock solution 10 times, 20 times, 100 times or more), that is, each of the small molecules prepared in Example 1 above. Take 1ml or 0.001g to 0.200g of alginate and low molecular weight fucoidan, dissolve in the medium, sterilize by UV treatment for 1 hour on a clean bench, dilute from 10,000ppm and inoculate 50μ of each active bacterium. The cell concentration over time was also measured at the minimum concentration (MIC) with antimicrobial activity of each sample. In addition, 20 μl of Seed Culture (species culture) incubated for 24 hours was smeared in a solid medium (Agar 20g / L medium added to the liquid medium) and cultured for 24 hours, and colonies were confirmed. In addition, the anaerobic culture in Table 5 was carried out in the same manner as the aerobic culture, but can be supplied to the mixed gas (80% N ₂ , 10% CO ₂ , 10% H ₂ ) in the anaerobic chamber by injecting the mixed gas in the anaerobic chamber The antimicrobial activity was investigated by the same method as aerobic culture, except that the cells were inoculated and cultured using a gas pack anaerobic system (Difco). In addition, ³⁾ LB medium is a medium containing 5g / L yeast extract, 10g / L tryptone and 10g / L sodium chloride, ⁴⁾ BHI is 30g / L Brain Heart Infusion (BD 237500), ^{5 )} TSB represents 35 g / L Tryptic Soy medium (BD 236950). The antimicrobial test results of Preparation Examples 1 to 40 of the low molecular weight alginate prepared in Example 1 according to the above method are shown in Table 6 below, and the antimicrobial test results of Preparation Examples 71 to 75 among the preparation examples of the low molecular weight fucotidan. Are shown in FIGS.

TABLE 6

Minimum concentration (MIC) measurement result with antimicrobial activity of low molecular weight alginate

* Measured at concentrations of 10,000 ppm

As a result, as shown in Table 6, in the case of the low-molecular alginate prepared by varying the conditions in the present invention, except for Preparation Examples 1 to 7 to all have the antibacterial activity against Escherichia coli, staphylococcus and acne bacteria In particular, in the case of Preparation 19 was found to have antimicrobial activity even at a very low concentration of 400ppm. In addition, in the case of low-molecular fucoidan was carried out antimicrobial activity test for Preparation Examples 71 to 75, this also showed that it has an antimicrobial activity against both E. coli, Staphylococcus aureus and acne bacteria, in particular for E. coli and acne Growth inhibition concentration (MIC) was found to be 10,000 ppm and Staphylococcus MIC was found to be 5,000 ppm.

Therefore, the results showed that the low molecular weight alginate and low molecular fucoidan according to the present invention had antibacterial activity against harmful strains of Escherichia coli, Staphylococcus aureus, and acne, and especially at low concentrations. I could see that.

<3-2> Antimicrobial Activity of Low Molecular Weight Alginate and Fucoidan

Furthermore, Escherichia coli , Propionibacterium acnes and Escherichia coli during the treatment of the sample to measure the antimicrobial properties of the low molecular weight alginate and low molecular weight fucoidan prepared in Example 1 (sample) with time. The cell concentration of Staphylococcus aureus was measured using UV wavelength (λ = 600 nm). At this time, the concentration of the sample was carried out in the MIC, samples for comparison were carried out at a constant concentration. The results for each of the prepared low molecular weight alginates are shown in Tables 7-9, and the results for low molecular weight fucoidan are listed in Tables 10-12 below.

TABLE 7

Cell concentration for small molecule alginate against E. coli (λ = 600 nm, 2,000 ppm)

Production Example

time 2 7 12 17 4 9 14 19 A-2.5-HP0-5 A-2.5-HP1.5-5 A-2.5-HP3.0-5 A-2.5-HP4.5-5 A-2.5-HP0-15 A-2.5-HP1.5-15 A-2.5-HP3.0-15 A-2.5-HP4.5-15 0 0 0 0 0 0 0 0 0 12 1.245 1.312 0.124 0.098 0.098 0.121 0.045 0.078 24 1.878 1.789 0.408 0.289 1.012 0.149 0.123 0.134 36 2.645 2.435 0.577 0.319 2.112 0.432 0.213 0.204 48 2.876 2.765 0.832 0.397 2.478 0.645 0.312 0.289 60 2.967 2.845 1.478 0.509 2.735 0.897 0.402 0.315

Production Example

time 22 27 32 37 24 29 34 39 A-1-HP
0-5 A-1-HP
1.5-5 A-1-HP
3.0-5 A-1-HP
4.5-5 A-1-HP
0-15 A-1-HP
1.5-15 A-1-HP
3.0-15 A-1-HP
4.5-15 0 0 0 0 0 0 0 0 0 12 1.245 1.312 0.187 0.107 1.145 1.012 0.274 0.102 24 1.878 1.789 0.432 0.324 1.658 1.789 0.412 0.313 36 2.645 2.435 0.623 0.389 2.325 2.135 0.598 0.375 48 2.876 2.765 0.823 0.423 2.678 2.123 0.845 0.412 60 2.967 2.845 1.545 0.612 2.660 2.345 1.412 0.524

[Table 8]

Cell concentration of small molecule alginate against staphylococci (λ = 600 nm, 2,000 ppm)

Production Example

time 2 7 12 17 4 9 14 19 A-2.5-HP0-5 A-2.5-HP1.5-5 A-2.5-HP3.0-5 A-2.5-HP4.5-5 A-2.5-HP0-15 A-2.5-HP1.5-15 A-2.5-HP
3.0-15 A-2.5-HP
4.5-15 0 0 0 0 0 0 0 0 0 12 1.323 1.423 0.224 0.125 0.145 0.215 0.095 0.068 24 1.745 1.896 0.452 0.345 1.432 0.292 0.189 0.145 36 2.443 2.532 0.598 0.423 2.145 0.539 0.256 0.223 48 2.745 2.813 0.924 0.455 2.678 0.748 0.378 0.295 60 2.967 2.987 1.512 0.612 2.871 0.989 0.443 0.367

Production Example

time 22 27 32 37 24 29 34 39 A-1-HP
0-5 A-1-HP
1.5-5 A-1-HP
3.0-5 A-1-HP
4.5-5 A-1-HP0-15 A-1-HP
1.5-15 A-1-HP
3.0-15 A-1-HP
4.5-15 0 0 0 0 0 0 0 0 0 12 1.245 1.467 0.235 0.145 1.323 1.117 0.302 0.102 24 1.878 1.845 0.412 0.333 1.756 1.867 0.444 0.313 36 2.645 2.512 0.608 0.404 2.432 2.354 0.623 0.375 48 2.876 2.866 0.945 0.545 2.756 2.645 0.887 0.412 60 3.167 2.845 1.545 0.687 2.867 2.798 1.412 0.524

TABLE 9

Cell concentration (λ = 600 nm, 2,000 ppm) for P. acnes of small molecule alginate

Production Example

time 2 7 12 17 4 9 14 19 A-2.5-HP
0-5.0 A-2.5-HP
1.5-5 A-2.5-HP
3.0-5 A-2.5-HP
4.5-5 A-2.5-HP
0-15 A-2.5-HP
1.5-15 A-2.5-HP
3.0-15 A-2.5-HP
4.5-15 0 0 0 0 0 0 0 0 0 12 0.745 0.345 0 0 0.645 0.624 0 0 24 1.145 0.761 0.125 0 1.080 1.064 0 0 36 2.565 1.251 0.287 0 2.345 1.345 0.112 0 48 2.745 1.675 0.425 0.085 2.765 1.745 0.213 0 60 3.002 1.864 1.669 0.120 2.954 2.045 0.304 0.089

Production Example

time 22 27 32 37 24 29 34 39 A-1-HP
0-5 A-1-HP
1.5-5 A-1-HP
3.0-5 A-1-HP
4.5-5 A-1-HP
0-15 A-1-HP
1.5-15 A-1-HP
3.0-15 A-1-HP
4.5-15 0 0 0 0 0 0 0 0 0 12 0.423 0.346 0 0 0.214 0 0 0 24 0.605 0.523 0.231 0 0.470 0 0 0 36 1.245 0.897 0.438 0.234 1.034 0 0 0 48 2.345 1.243 0.756 0.453 1.546 0 0 0.087 60 3.081 1.622 1.318 1.051 2.816 0.097 0.076 0.103

TABLE 10

Cell concentration (λ = 600 nm, 10,000 ppm) for E. coli of fucoidan

Production Example

time 71 72 73 74 75 Native fucoidan F-1-HP3.0-2.5 F-1-HP3.0-5.0 F-1-HP3.0-10.0 F-1-HP3.0-15.0 F-1-HP3.0-20.0 0 0 0 0 0 0 0 12 1.648 0.012 0.045 0.035 0.028 0.085 24 1.950 0.856 0.695 0.100 0.098 0.098 36 2.455 1.845 0.745 0.145 0.124 0.115 48 2.645 2.125 1.475 0.478 0.198 0.175 60 2.745 2.675 2.025 1.012 0.412 0.397

TABLE 11

Cell density (λ = 600nm, 5,000ppm) for Staphylococcus aureus of fucoidan (S. Aureus)

Production Example

time 71 72 73 74 75 Native fucoidan F-1-HP3.0-2.5 F-1-HP3.0-5.0 F-1-HP3.0-10.0 F-1-HP3.0-15.0 F-1-HP3.0-20.0 0 0 0 0 0 0 0 12 1.545 0.087 0.025 0.045 0.075 0.075 24 1.798 1.565 0.160 0.105 0.087 0.081 36 2.345 2.785 0.245 0.178 0.135 0.125 48 3.085 3.145 0.375 0.295 0.187 0.172 60 2.875 2.975 0.815 0.545 0.387 0.342

TABLE 12

Cell density (λ = 600nm, 10,000ppm) for the fucoidan acne bacteria (P. acnes)

Production Example

time 71 72 73 74 75 Native fucoidan F-1-HP3.0-2.5 F-1-HP3.0-5.0 F-1-HP3.0-10.0 F-1-HP3.0-15.0 F-1-HP3.0-20.0 0 0 0 0 0 0 0 12 0.678 0.045 0.012 0.045 0.058 0.067 24 1.343 0.098 0.075 0.068 0.061 0.062 36 1.623 0.185 0.154 0.148 0.132 0.098 48 2.735 0.865 0.287 0.265 0.232 0.105 60 2.545 1.875 0.745 0.623 0.585 0.304

As a result, as shown in Tables 7 to 12, the low molecular alginate and low molecular fucoidan were found to decrease the growth rate of the proliferating cells with a longer treatment time. Therefore, the present inventors found that when the low molecular weight alginate and the low molecular fucoidan were treated at the minimum growth inhibition concentration (MIC), the antimicrobial activity also increased as the treatment time increased.

<3-3> Antimicrobial Activity of Low Molecular Silver-Alginate

The antimicrobial activity of the low molecular weight silver-alginate prepared in Example 1 was investigated. Silver (Ag) content is more than 35%, the size of the silver alginate particles was measured MIC using a sample in the range of 0.1 ~ 3.4㎛, the results are shown in Table 13 and FIG.

TABLE 13

Antimicrobial Investigation of Low Molecular Silver-Alginate

Strain name Low Molecular Silver Alginate Solution (ppm) MIC (ppm) 700 350 175 Escherichia coli Do not multiply Do not multiply Multiply 350 Staphylococcus Do not multiply Do not multiply Multiply 350 Acne Do not multiply Do not multiply Multiply 350

As a result of the antimicrobial investigation, as shown in Table 13 and FIG. 4, the low molecular weight silver-alginate according to the present invention was found to have antimicrobial activity against the strains above MIC 350ppm concentration against E. coli, Staphylococcus aureus, and acne cocci. In particular, the concentration of 700ppm was not able to proliferate all three types of strains. On the other hand, at 175 ppm concentration, the strains appeared to grow somewhat. Therefore, the results showed that the low molecular weight silver-alginate had antimicrobial activity, and in particular, MIC for E. coli, staphylococcus and acne bacterium was found to be 350ppm.

<Example 4>

Analysis of Radical Scavenging and Cytotoxic Activity of Low Molecular Alginate and Fucoidan

<4-1> radical scavenging activity

Analysis of hydroxyl radicals is described by N. Smirnoff and Q.J. Method known by Cumbes

(Phytochemistry, 28 (4), 1057, 1989), i.e. , 25mM FeSO4, 2mM sodium salicylate, 6mM hydrogen peroxide in each sample was balltacked at 37 ° C for 1 hour and then absorbance at 510nm. The scavenging activity of the radical on the sample was measured. At this time, the inhibition rate of the radical was calculated by the following equation. In addition, as the sample, low molecular weight alginate, silver-alginate, polymer alginate and fucoidan prepared in Example 1 were used for each concentration, and the results are shown in Tables 14, 15, and FIGS. 5 to 8.

Inhibition Rate (%) = (A-B) / A × 100

(A is the control group, B is the experimental group)

[Table 14]

Absorbance of Silver-Alginate, Polymer Alginate and Low Molecular Alginate (510 nm)

Addition amount
(mg) BK Silver-alginate
(SA) Low Molecular Alginate
(LMWA) Polymer Alginate
(HMWA) 1.0 0.419 0.412 0.298 0.385 2.0 0.419 0.410 0.231 0.309 4.0 0.419 0.389 0.170 0.230 6.0 0.419 0.351 0.155 0.170 8.0 0.419 0.285 0.139 0.149

Addition amount
(mg) BK LMWA 1
(A-2.5-HP4.5-5) LMWA 2
(A-2.5-HP1.5-5) LMWA 3
(A-2.5-HP0-20) 1.0 0.432 0.298 0.291 0.301 2.0 0.432 0.231 0.228 0.258 4.0 0.432 0.170 0.163 0.185 6.0 0.432 0.155 0.149 0.166 8.0 0.432 0.139 0.134 0.128

TABLE 15

  Absorbance of Silver-Alginate, Low Molecular Alginate, Polymer Alginate and Fucoidan

Addition amount (mg) BK Low Molecular Alginate Polymer Alginate Silver-alginate Fucoidan 1.0 2.160 1.490 1.925 2.125 2.100 2.0 2.160 1.155 1.545 2.000 1.975 4.0 2.160 0.850 1.150 1.800 1.855 6.0 2.160 0.775 0.850 1.655 1.820 8.0 2.160 0.695 0.745 1.435 1.750

As a result, as shown in Table 14 and 15 and Figures 5 to 8, the radical scavenging activity showed a tendency to increase with the addition amount of alginate and fucoidan, in the case of silver-alginate, the scavenging activity of the radical is alginate and It was found to be relatively lower than fucoidan, but silver-alginate was also found to have a radical scavenging activity.

<4-2> Cytotoxicity Assay

<4-2-1> Cell culture method

Keratinocyte cell lines (HaCaT) were incubated in 37 ° C., 5% CO ₂ and in a humidified incubator (Sanyo). In this case, DMEM (dulbecco's modified eagle medium, Invitrogrn) medium containing 10% fetal bovine serum (FBS, GibcoBRL) and antibiotics (antibiotic-antimycotic, GibcoBRL) was heat-treated at 56 ° C for 30 minutes.

<4-2-2> Cytotoxicity Test

Cytotoxicity tests showed that an enzyme called succinic dehydrogenase in the mitochondria of living cells reduced MTT [3- (4,5-dimethyl-2-yl) -2, 5-diphenyl tetrazolium bromide]. On the basis of the formazan (formazan) can be formed by measuring the concentration of the formed formazan was used MTT method to measure the number of cells survived (Pieters et al. 1988). Cancer cells with normal metabolic processes reduce yellow water-soluble MTT tetrazolium to purple water-insoluble MTT formazan by the dehydrogenase action of mitochondria. The absorbance of MTT formazan peaks at a wavelength of 540 nm, and the absorbance measured at this wavelength reflects the concentration of living and metabolically active cells. Through this method, in this example, the resistant cell lines cultured without drugs in a 96-well microplate for 3 days were diluted to an appropriate concentration, and 90 µl of the cell suspension was added. After 18 hours, 10 μl of the drug to be measured was added. At this time, PBS (phosphate buffered saline) was used instead of the drug as a control cell, and only the culture medium was used as a blank. Shake well, incubate in CO ₂ incubator for 3 days, and add all wells to 10 μl of MTT solution (5 mg / ml PBS, Sigma) and incubate another 4-5 hours at 37 ° C and 5% CO ₂ . MTT was reduced. After 5 hours, 80 μl was removed from each well, and then 150 μl DMSO was added and the resulting formazan crystal was dissolved while stirring for 10 minutes. The absorbance was measured at 540 nm using a microplate reader (Bio-Tek, USA). The results are shown in Table 16 and FIG. 9.

TABLE 16

Cytotoxicity Measurements of Low Molecular Alginate, Fucoidan and Silver-Alginate

sample A-2.5-HP4.5-20
(4.5% hydrogen peroxide) Low Molecular Silver-Alginate Polymer Fucoidan Concentration (ug / ml) Absorbance Absorbance Absorbance 1000 0.039 0.044 0.026 0.249 0.288 0.3 0.07 0.033 0.017 200 0.257 0.166 0.095 0.277 0.423 0.53 0.174 0.19 0.199 40 0.4 0.363 0.397 0.555 0.51 0.604 0.408 0.38 0.468 8 0.596 0.63 0.519 0.587 0.617 0.482 0.593 0.668 0.696 1.6 0.633 0.507 0.572 0.705 0.638 0.545 0.556 0.523 0.65 0.32 0.525 0.667 0.598 0.652 0.624 0.597 0.608 0.681 0.625 0.064 0.607 0.445 0.498 0.515 0.563 0.601 0.532 0.556 0.536 Control 0.557 0.367 0 0.416 0.573 0.544 0.565 0.538 0.523

sample F-2.5-HP0-20
(No hydrogen peroxide) A-2.5-HP0-20
(No hydrogen peroxide) A-2.5-HP3.0-20
(Hydrogen peroxide 3.0%) Concentration (ug / ml) Absorbance Absorbance Absorbance 1000 0.325 0.392 0.318 0.249 0.288 0.3 0.07 0.033 0.017 200 0.266 0.5 0.394 0.277 0.423 0.53 0.174 0.19 0.199 40 0.522 0.434 0.423 0.555 0.51 0.604 0.408 0.38 0.468 8 0.42 0.366 0.557 0.587 0.617 0.482 0.593 0.668 0.696 1.6 0.285 0.383 0.365 0.705 0.638 0.545 0.556 0.523 0.65 0.32 0.531 0.363 0.426 0.652 0.624 0.597 0.608 0.681 0.625 0.064 0.384 0.3 0.409 0.515 0.563 0.601 0.532 0.556 0.536 Control 0.486 0.378 0.532 0.416 0.573 0.544 0.565 0.538 0.523

As a result, as shown in Table 16 and Figure 9, the polymer alginate, fucoidan and low-molecular silver alginate at a solution concentration of 1000 ppm was observed to be non-cytotoxic, low-molecular alginate and fucoidan without hydrogen peroxide was also found to be not cytotoxic. appear. For reference, in FIG. 9, the darker the color, the less toxic, and the lighter or darker the color.

Therefore, the present inventors through the above results, since the low molecular alginate, low molecular fucoidan and low molecular silver-alginate according to the present invention is not toxic to cells, when used in the manufacture of cosmetics, it is possible to prepare a cosmetic having excellent antibacterial activity while being safe for skin. Could see.

< Example  5>

Preparation of cosmetics having antimicrobial activity according to the present invention

Preparation of cosmetics with excellent antimicrobial properties according to the present invention was prepared by using the low-molecular alginate, low-molecular fucoidan and low-molecular silver alginate alone or in combination of the present invention, according to the composition ratio of each component shown in Table 17 Was prepared. All materials and containers were disinfected and the manufacturing process was performed on a clean bench. First, the oil layer (A) is weighed in a vessel 1 and boiled at 70 ° C., then the water phase layer (B) is weighed and dissolved in another vessel 2, and then dissolved, and allantoin and triclosan are added to the additive C. And ascrobic acid (ascrobic acid) was added, and when it is confirmed that the melted additive (E) was dissolved. At this time, the additive was heated to a temperature and time only enough to melt, and not to be heated for as long as possible. Thereafter, the oil layer (A) was added to the water phase layer (B) of the container 2 while stirring, and the mixture was mixed with a hand blender to make an emulsified state. Then, when the temperature was lowered to 40 ° C., the remaining ingredients of the additive (C) were mixed rapidly and mixed into the sterilized container.

TABLE 17

Composition and ratio (unit%) of the essence cosmetic prepared in the present invention

ingredient Application example 1 Application example 2 Application example 3 Application example 4 Application example 5 Application Example 6 Application example 7 Application example 8 Application example 9 One Avocado oil Oh first floor
(A) 5 5 5 5 5 5 5 5 5 2 Emulsifying wax 3 3 3 3 3 3 3 3 3 4 Rose water Water layer
(B) 40 40 40 40 40 40 40 40 40 5 Tea tree floral water Up to 100 Up to 100 Up to 100 Up to 100 Up to 100 Up to 100 Up to 100 Up to 100 Up to 100 6 Aloevera gel 12 12 12 12 12 12 12 12 12 7 Vegetable glycerin additive
(C) 2 2 2 2 2 2 2 2 2 8 Vitamin e One One One One One One One One One 9 Olive liquid One One One One One One One One One 10 Polysorbate One One One One One One One One One 11 Hyaluronic acid One One One One One One One One One 12 Allantonin 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 13 Triclosan 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 14 Phenoxyethanol One One One One One One One One One 15 Ascorbic acid One One One - - One - - - 16 Ethanol - - - - 1.5 - - - - 17 Chamomile E / O (D) One One One One One One One One One 18 Rose geranium E / O 2 2 2 2 2 2 2 2 2 19 Polymer Alginate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 (E) 20 Low Molecular Alginate - 0.5 - - - 0.5 0.5 - 0.5 21 Polymer Fucoidan - - 0.5 - - 0.5 - 0.5 0.5 22 Low Molecular Fucoidan 0.5 0.5 - 0.5 0.5 - 0.5 - - 23 Low Molecular Silver-Alginate - - - 0.035 0.035 - 0.035 0.035 0.035

< Example  6>

Investigation of antimicrobial activity of manufactured cosmetics

Antibacterial test for E. coli, Staphylococcus aureus, acne bacteria and anthrax were performed on the essence cosmetics of Examples 1 to 9 prepared by varying the content and composition of each component in Example 5. The antimicrobial test was conducted in the same manner as in Example <3-1> at a concentration of 20,000 ppm with respect to the sample of the cosmetic prepared above, the results are shown in FIG.

As a result, as shown in Figure 10, in the case of cosmetics prepared containing the low molecular alginate, low molecular fucoidan and low molecular silver-alginate according to the present invention, the antibacterial activity against colon coli, staphylococci, acne and anthrax In particular, in the case of cosmetics containing low molecular weight silver-alginate at a MIC concentration of 350 ppm, the antibacterial activity of acne bacteria including anthrax was excellent.

So far I looked at the center of the preferred embodiment for the present invention. Those skilled in the art will appreciate that the present invention can be implemented in a modified form without departing from the essential features of the present invention. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is shown in the appended claims rather than the foregoing description, and all differences within the scope will be construed as being included in the present invention.

1 is a photograph showing the antimicrobial activity against Escherichia coli when the low molecular weight fucoidan according to the present invention is treated at each concentration.

Figure 2 is a photograph showing the antimicrobial activity against Staphylococcus aureus when the low-molecular fucoidan according to the present invention treated for each concentration.

Figure 3 is a photograph showing the antimicrobial activity against acne bacteria when the low-molecular fucoidan according to the present invention treated for each concentration.

Figure 4 is a photograph showing the antimicrobial activity against Escherichia coli, Staphylococcus aureus and acne bacteria at each concentration for the low molecular weight silver-alginate solution according to the present invention.

Figure 5 shows a graph of the radical scavenging activity of the low molecular weight alginate according to the present invention by measuring the absorbance of the hydroxy radicals.

FIG. 6 is a graph showing the antioxidant activity of the low molecular silver-alginate, the high molecular silver-alginate and the silver-alginate.

7 is a graph showing the antioxidant activity of the low molecular weight alginate (Preparation Example 17, 7 and 5 prepared in one embodiment of the present invention).

8 is a graph showing the antioxidant activity of the low-molecular weight silver-alginate, high molecular silver-alginate, silver-alginate and fucoidan according to the present invention.

Figure 9 is a photograph showing the results of the cytotoxicity test for the low molecular weight silver-alginate, high molecular silver-alginate, silver-alginate and fucoidan according to the present invention by MTT analysis.

10 is a comparison of the antimicrobial activity against E. coli, Staphylococcus aureus, acne and anthrax for essence cosmetics prepared by combining low molecular weight alginate, low molecular weight fucoidan and low molecular weight silver alginate alone or in accordance with one embodiment of the present invention. It is a photograph.

Claims (6)

0.01-1.0% (w / v) of at least one selected from the group consisting of a low molecular weight alginate having a molecular weight of 150,000 Da or less, a low molecular weight fucoidan having a molecular weight of 35,000 Da or less and a low molecular weight silver-alginate having a particle size of 0.1 to 3.4 µm Antimicrobial cosmetic composition comprising). The method of claim 1, The low molecular weight alginate or low molecular fucoidan is an antimicrobial cosmetic composition, characterized in that prepared by the step of irradiating an electron beam of 1.0 to 3.0 MeV in an aqueous solution containing 0.1 to 2.0% by weight of alginate or fucoidan. The method of claim 1, The low molecular weight silver-alginate is added to the aqueous solution containing 0.1 to 2.0 wt% of alginate in an amount of 1.5 to 4.5 wt% of hydrogen peroxide, and then irradiated with an electron beam of 1.0 to 3.0 MeV to the mixed solution to which the hydrogen peroxide is added. Antibacterial cosmetic composition, characterized in that prepared by the step of reacting by adding a silver nitrate (AgNO ₃ ) solution to recover the precipitate. The method of claim 1, The antimicrobial cosmetic composition is characterized in that it has an antimicrobial activity and radical scavenging activity against a strain selected from the group consisting of E. coli, staphylococcus, acne bacteria and anthrax. The method of claim 1, The low-molecular alginate, low-molecular fucoidan and low-molecular silver-alginate have a minimum growth inhibition concentration (MIC) for the strains of 400 to 10000 ppm, 5000 to 10000 ppm, and 300 to 400 ppm, respectively. The method of claim 1, The composition includes skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutrition cream, moisturizing cream, hand cream, foundation, essence, nutrition essence, pack, soap, cleansing foam Antibacterial cosmetic composition, characterized in that the formulation of any one selected from the group consisting of cleansing lotion, cleansing cream, body lotion and body cleanser.

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