KR20190067442A - Cosmetic composition comprising oysters, sea cucumber, sea squirt, Aplysia kurodai and buccinum striatissimum - Google Patents

Abstract

The present invention relates to a functional cosmetic composition comprising oysters, sea squirts, sea cucumbers, sea cucumbers, Aplysia kurodai and Buccinum striatissimum as an active ingredient. The composition according to the present invention may be useful as a cosmetic composition by having an effect of significantly improving the antioxidant activity, skin whitening effect, skin moisturizing effect and skin wrinkle reduction effect without skin irritation.

Description

[0001] The present invention relates to a cosmetic composition comprising oysters, sea cucumber, sea squirt, Aplysia kurodai and buccinum striatissimum containing oysters, sea urchins, sea cucumbers,

The present invention relates to a functional cosmetic composition comprising oysters, sea urchins, sea cucumbers, small parts and nasal cavities as active ingredients.

Aging is a functional, structural, and biochemical change that occurs throughout the whole body of cells and body tissues. Human skin has decreased hormone secretion over time and decreased function and activity of immune cells, resulting in intrinsic aging resulting from decreased biosynthesis of biocompatible proteins. In addition, dry skin, pollutants and ultraviolet light The skin is thinned, the wrinkles are increased, the elasticity is reduced, and aging phenomena such as spots, freckles and black blotch occur.

In particular, photooxidative skin damage by ultraviolet light is mediated by reactive oxygen species (ROS), leading to antioxidant depletion, lipid peroxidation, protein oxidation and oxidative damage of DNA. These ROS disrupt skin antioxidant defense membranes composed of enzymatic and non-enzymatic antioxidants, oxidative deformation of biomolecules, damage of skin barrier, chain breaks such as collagen and hyaluronic acid, and wrinkle formation by abnormal cross-linking Accelerate skin aging.

 When human skin is exposed to external stimuli, especially ultraviolet rays, collagenase increases its activity to decompose the collagen of the skin and reduce the content of collagen. Collagenase is known to cleave collagen and turn it into an irregular structure, which causes the elasticity of the skin to fall and wrinkles.

The skin color of a person is determined by the amount of melanin, carotene and hemoglobin. Melanin is the most important factor. Melanin is synthesized in the melanocyte, a pigmented cell in the basal layer of the skin (epidermis), and transformed into keratinocyte to show the color of human skin. Unusually low production of melanin causes skin lesions such as Vitiligo. On the contrary, overproduction is one of the major concerns of middle-aged women, forming spots and freckles, and is also closely related to skin cancer.

The whitening effect will be described in more detail as follows. Melanin is produced by enzymes and nonenzymatic oxidation of tyrosine in cells called melanocytes, which are found in the basal layer of skin epidermis, and transformed into keratinocytes constituting the epidermis. Important enzymes involved in the biosynthesis of melanin include tyrosinase, tyrosinase-related protein 1 and dopachrome tautomerase. In addition to tyrosinase, these two enzymes attract attention, but the enzyme that plays a crucial role in the synthesis of melanin is tyrosinase. The first step in the biosynthesis of melanin is caused by tyrosinase. Tyrosine is converted to tyrosine by tyrosinase and this tyrosine is made into Dopa-quinone. The next step in these two reactions is known to be possible by nonenzymatic reactions, and tyrosinase alone is known to be able to produce melanin.

Accordingly, the present inventors have completed the present invention by confirming the antioxidative activities of oysters, sea urchins, sea cucumbers, small spots, nose pals, kelp and kotatsu, skin whitening effect, skin moisturization enhancement and skin wrinkle improvement.

Korean Patent No. 10-1640799

It is an object of the present invention to provide a cosmetic composition having antioxidative activity, which comprises at least one selected from the group consisting of oyster, astringent skin, sea tangle, and locust.

Another object of the present invention is to provide a cosmetic composition for whitening skin, which comprises at least one member selected from the group consisting of oysters, mace, kelp, and the like.

Another object of the present invention is to provide a cosmetic composition for enhancing skin moisturizing, which comprises at least one member selected from the group consisting of oysters, mace, kelp, and the like.

Another object of the present invention is to provide a cosmetic composition for improving skin wrinkles, which comprises at least one member selected from the group consisting of oysters, mace, kelp, and the like.

In order to achieve the above object,

The present invention includes at least one member selected from the group consisting of oysters, kelp, kelp, and locusts, wherein the oysters and the oysters are hydrolyzed with protein hydrolytic enzymes after being ground in water at 80-100 ° C, The present invention provides a cosmetic composition having an antioxidative activity, characterized in that kelp and kelp are dried and then ground and extracted with an organic solvent.

In addition, the present invention includes at least one member selected from the group consisting of oysters, sweet potatoes, seaweeds, and spores, wherein the oysters and the oysters are hydrolyzed with protein hydrolyzing enzyme after being ground in water at 80-100 ° C, And the kelp and kelp are dried and then ground, and extracted with an organic solvent. The cosmetic composition for skin whitening is provided.

Further, the present invention includes at least one member selected from the group consisting of oysters, cutulas, kelp, and small groups, wherein the oysters and the oysters are hydrolyzed with protein hydrolytic enzymes after being ground in water at 80-100 ° C, , And the kelp and kelp are dried, ground, and extracted with an organic solvent before use.

Furthermore, the present invention includes at least one member selected from the group consisting of oysters, cutulas, kelp, and small groups, wherein the oysters and the oysters are hydrolyzed with protein hydrolytic enzymes after being ground in water at 80-100 ° C, And the kelp and kelp are dried and then ground, and extracted with an organic solvent. The cosmetic composition for improving skin wrinkles is provided.

The composition according to the present invention may be useful as a cosmetic composition because it has the effect of significantly improving the antioxidant activity, skin whitening effect, skin moisturizing effect and skin wrinkle reducing effect without skin irritation.

Hereinafter, the present invention will be described in detail.

Cosmetic composition

The present invention provides a cosmetic composition having an antioxidative activity comprising at least one member selected from the group consisting of oysters, kotatsu, kelp, and locusts.

The present invention also provides a cosmetic composition for skin whitening comprising at least one member selected from the group consisting of oysters, kotatsu, kelp, and locusts.

Further, the present invention provides a cosmetic composition for enhancing skin moisturization comprising at least one member selected from the group consisting of oysters, mace, kelp, and locusts.

Further, the present invention provides a cosmetic composition for improving skin wrinkles, comprising at least one selected from the group consisting of oysters, kotatsu, kelp, and locusts.

In one embodiment of the present invention, the oyster and the oyster are crushed and then hydrolyzed with a protein hydrolyzing enzyme in water at 80-100 ° C. The kelp and kotai are dried and then ground and extracted with an organic solvent .

In one embodiment of the present invention, the organic solvent comprises one or more compounds selected from the group consisting of methanol, ethanol, butanol, ethyl acetate, acetonitrile, acetone, 1,3-butylene glycol, And ethanol can be preferably used, but not limited thereto.

In one embodiment of the present invention, the composition may be used in an amount of 1.75-2.25 parts by weight, 0.25-0.75 parts by weight of kelp, and 0.25-0.75 parts by weight, based on 2 parts by weight of the oyster, preferably 2 parts by weight As a standard, 2 parts by weight of menthol, 0.5 part by weight of kelp, and 0.5 part by weight of the composition can be used.

Also, in one embodiment of the present invention, 1.75-2.25 parts by weight, 1.75-2.25 parts by weight of kelp, and 0.25-0.75 parts by weight may be used based on 0.5 parts by weight of the oyster, 2 parts by weight of menthol, 2 parts by weight of sea tangle, and 0.5 part by weight of a colony.

The term "whitening" of the present invention refers to inhibiting, inhibiting or alleviating hyperpigmentation of the skin. Hyperpigmentation of the skin includes freckles, stains, hyperpigmentation after exposure to ultraviolet light, hyperpigmentation after inflammation, aging black spots, brown spots or black spots.

The cosmetic composition according to the present invention may be used as a cosmetic composition for external use such as an ointment for external use, a cream, a softener, a nutritional lotion, a pack, an essence, a hair tonic, a shampoo, a rinse, a hair conditioner, a hair treatment, a gel, a skin lotion, a skin softener, It consists of lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutrition cream, moisturizing cream, hand cream, foundation, nutrition essence, sunscreen, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser ≪ RTI ID = 0.0 > and / or < / RTI > The composition of each of these formulations may contain various kinds of bases and additives necessary for formulation of the formulation, and the kinds and amounts of these ingredients can be easily selected by those skilled in the art.

When the formulation of the present invention is a paste, cream or gel, animal fiber, plant fiber, 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.

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

When 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 is selected from 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, linolenic derivatives or ethoxylated glycerol fatty acid esters.

The formulation of the present invention may further contain an excipient including a fluorescent substance, a fungicide, a hygroscopic substance, a humidifier, a fragrance, a fragrance carrier, a protein, a solubilizing agent, a sugar derivative, a sunscreen, a vitamin, .

When the cosmetic composition is formulated into a pharmaceutical or cosmetic composition as described above, it may contain known excipients applicable to the skin acting as a carrier for the active ingredient. When formulating into pharmaceuticals, reference is made to the disclosure of [Remington ' s Pharmaceutical Science, Mack Publishing Company, Easton PA], and in formulation into cosmetics, International cosmetic ingredient dictionary, 6th ed., The cosmetic, Toiletry and Fragrance Association, Inc., Washington, 1995). Which are incorporated herein by reference.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are illustrative of the present invention, and the present invention is not limited by the following examples.

≪ Example 1 > Production of raw materials

The oysters were incubated at 100 ° C for 10-30 minutes and then treated with Protamex at 50 ° C for 1 hour and neutrase at 60 ° C for 1 hour. The starch was hydrolyzed at 60 ℃ for 3 hours by adding alkalase to the starch. The starch was added to water at 100 ℃ for 10-30 min.

After thoroughly rinsing the nostrils, add the same amount of water and boil for 30 minutes at 105 ° C. Boil the noodles with the chopper and mix with the juice. Add 1% of the dextrozymes of the cooked meat and hydrolyze at 61 ℃ for 24 hours. After the hydrolysis, the mixture is inactivated at 100 DEG C for 30 minutes. Filter through a 200 mesh sieve and concentrate to 30 Brix. Add 3 times the amount of the concentrate to the Overnight. Thereafter, the precipitate is recovered to completely remove ethanol. The lyophilized powder was dissolved in water to prepare the raw material.

The sea urchin was extracted with hot water and then mucopolysaccharide was precipitated and used as a raw material.

The extracts of various seaweeds (kelp, mackerel, hearing, and seaweed) and seaweeds (top, hearing, mulberry, kelp, kelp extract) were dried and then ground, and the active ingredient was extracted by adding 10 times of ethanol to the weight of seaweed, , And the remaining extract was dissolved in water and butylene glycol and used as a raw material.

<Experimental Example 1> Evaluation of DPPH radical scavenging activity

DPPH assay was performed according to the method used by Yoshida et al. (1989). 100 μl of deionized water is added to the 96-well plate, and 100 μl of sample diluted with deionized water is added to the test group. Add 100 μl of ethanol into all wells and add 50 μl of a solution of DPPH (0.5 mM 1,1-diphenyl-2-picrylhydrazyl (DPPH) / ethanol). After reacting at room temperature for 30 minutes at 25 ° C, the absorbance was measured at 517 nm. As a solvent control, the radical scavenging ability of the control group was expressed as a percentage.

DPPH radical scavenging ability (%) seabird* 28.2 ± 7.8 Top 33.4 ± 0.2 ear 2.5 ± 5.0 oyster 3.3 ± 1.9 Sea cucumber 12.0 ± 6.2 Moth 31.7 ± 6.0 Kelp 38.3 ± 1.7 sea hare 0.0 ± 1.1 Koh Gong 92.1 ± 2.7 Moth 10.5 ± 1.9 sea squirt 20.7 ± 1.0

* Seaweed: Mixed extract of kelp, mackerel, hearing, potato

As shown in Table 1, DPPH radical scavenging activity of antioxidant activity was the highest at 92.1% of nodule, and DPPH radical scavenging activity of seaweed extract and kelp extract was the next highest.

<Experimental Example 2> Evaluation of ABTS radical scavenging ability

The ABTS assay method was modified by the method of Re et al. (1999). That is, 5 mL of 7 mM ABTS and 88 μl of 140 mM potassium persulfate are mixed, and then light is blocked at room temperature for 16 hours to form ABTS cations. This solution was then diluted with PBS to an absorbance value of 1.5 at 414 nm. 190 μl of the prepared diluted solution and 10 μl of the sample were mixed, reacted at room temperature for 6 minutes, and the absorbance was measured at 734 nm. As a solvent control, the radical scavenging ability of the control group was expressed as a percentage.

ABTS radical scavenging ability (%) seabird* 83.1 ± 3.0 Top 88.2 ± 13.0 ear 25.5 ± 2.9 oyster 97.6 ± 15.1 Sea cucumber 67.8 ± 6.5 Moth 50.0 ± 5.4 Kelp 52.4 ± 5.2 sea hare 51.3 ± 6.9 Koh Gong 76.9 ± 16.0 Moth 99.5 ± 8.3 sea squirt 97.9 ± 5.8

* Seaweed: Mixed extract of kelp, mackerel, hearing, potato

As shown in Table 2, ABTS radical scavenging ability was highest in the order of 99.5%, and in the order of 97.9% and 97.6%, respectively.

<Experimental Example 3> Evaluation of inhibitory activity against tyrosinase activity

The sample is dissolved in an appropriate solvent and diluted to a suitable concentration range to inhibit tyrosinase activity. Add 220 μl of 0.1 M phosphate buffer (pH 6.5), 20 μl of sample solution and 20 μl of mushroom tyrosinase (2 U / μl) in the test tube in this order. To this solution, 40 μl of a 1.5 mM tyrosine solution was added and reacted at 37 ° C. for 30 minutes. The absorbance is measured at 490 nm using an ELISA reader. Add 0.1 M phosphate buffer (pH 6.5) instead of the sample solution to the blank.

 a: Absorbance after reaction of blank sample

 b: Absorbance after reaction of the sample solution

 a ', b': Absorbance measured by replacing with tyrosinase buffer

Tyrosinase activity inhibition (%) seabird* - Top - ear - oyster - Sea cucumber - Moth - Kelp - sea hare - Koh Gong - Moth 81.1 ± 0.3 sea squirt -

* Seaweed: Mixed extract of kelp, mackerel, hearing, potato

As shown in Table 3, the result of the inhibition of the activity of thyronidase was 81.1%.

<Experimental Example 4> Evaluation of moisturizing efficacy

To measure the moisturizing effect, 1 ml of the sample was dipped in a cotton swab at 37 ° C in an incubator, and the water holding capacity was measured by measuring the weight per hour. The moisture content was measured with distilled water not containing the sample as a control, and the moisture content was measured with the lapse of time, and the weight was measured at intervals of 1 hour, 2 hours and 4 hours.

Moisturizng (%) seabird* 24.8 ± 0.7 Top 30.3 ± 4.7 ear 33.5 ± 2.8 oyster 41.7 ± 1.5 Sea cucumber 27.0 ± 5.3 Moth 40.4 ± 2.7 Kelp 56.6 ± 1.6 sea hare 45.2 ± 3.6 Koh Gong 45.7 ± 4.8 Moth 52.6 ± 1.9 sea squirt 53.4 ± 3.9

* Seaweed: Mixed extract of kelp, mackerel, hearing, potato

As shown in Table 4, seaweed was the highest at 56.6%, followed by sea urchin at 53.4%, seaweed at 52.2% and ko god at 45.7%.

The most effective oyster extract, colchic extract, gentian extract, and sea tangle extract were selected through Experimental Examples 1 to 4, and the activity of each of the oyster extracts, mushroom extracts, and sea tangle extracts was measured by varying the mixing ratios of the four extracts.

<Example 2> Formulation of oyster, kotai, kelp, and locust extract

Mixing Ratio (mL) oyster Moth Kelp sea hare Example 2-1 0.5 0.5 0.5 0.5 Example 2-2 0.5 0.5 0.5 2 Example 2-3 0.5 0.5 2 0.5 Examples 2-4 0.5 0.5 2 2 Example 2-5 0.5 2 0.5 0.5 Examples 2-6 0.5 2 0.5 2 Examples 2-7 0.5 2 2 0.5 Examples 2-8 0.5 2 2 2 Examples 2-9 2 0.5 0.5 0.5 Examples 2-10 2 0.5 0.5 2 Examples 2-11 2 0.5 2 0.5 Examples 2-12 2 0.5 2 2 Examples 2-13 2 2 0.5 0.5 Examples 2-14 2 2 0.5 2 Examples 2-15 2 2 2 0.5 Examples 2-16 2 2 2 2

<Experimental Example 5> Evaluation of ABTS radical scavenging ability according to the difference of the mixing ratio of extracts of oysters, kotatsu, kelp,

The ABTS radical scavenging ability according to the mixing ratio of Example 2 was measured. The ABTS assay method was modified by the method of Re et al. (1999).

ABTS radical scavenging activity (%) Example 2-1 84.1 ± 2.0 Example 2-2 77.1 ± 4.0 Example 2-3 72.6 ± 8.8 Examples 2-4 80.4 ± 8.8 Example 2-5 99.7 ± 14.1 Examples 2-6 99.8 ± 15.7 Examples 2-7 100.2 ± 39.8 Examples 2-8 99.9 ± 70.7 Examples 2-9 94.5 ± 24.9 Examples 2-10 94.8 ± 34.2 Examples 2-11 94.8 ± 16.6 Examples 2-12 91.8 ± 25.1 Examples 2-13 99.9 ± 25.0 Examples 2-14 99.7 ± 11.8 Examples 2-15 99.8 ± 39.0 Examples 2-16 99.7 ± 3.8

<Experimental Example 6> Evaluation of inhibitory activity of tyrosinase activity

In the same manner as in Experimental Example 3, the inhibitory activity of tyrosinase activity according to the mixing ratio of Example 2 was measured.

Tyrosinase (%) Example 2-1 46.8 ± 10.5 Example 2-2 31.5 ± 9.0 Example 2-3 20.6 ± 6.0 Examples 2-4 40.7 ± 9.9 Example 2-5 84.6 ± 6.1 Examples 2-6 73.0 ± 1.6 Examples 2-7 85.1 ± 11.5 Examples 2-8 82.1 ± 14.0 Examples 2-9 35.6 ± 5.0 Examples 2-10 36.0 ± 7.3 Examples 2-11 31.6 ± 0.5 Examples 2-12 26.1 ± 0.7 Examples 2-13 84.2 ± 2.8 Examples 2-14 82.6 ± 8.9 Examples 2-15 83.6 ± 3.8 Examples 2-16 80.7 ± 2.6

<Experimental Example 7> Measurement of moisturizing effect

The moisturizing effect according to the mixing ratio of Example 2 was measured in the same manner as in Experimental Example 4.

Moisturizng (%) Example 2-1 38.9 ± 3.0 Example 2-2 42.8 ± 3.3 Example 2-3 66.9 ± 18.3 Examples 2-4 54.2 ± 10.0 Example 2-5 47.9 ± 3.6 Examples 2-6 58.8 ± 10.3 Examples 2-7 67.6 ± 8.9 Examples 2-8 65.2 ± 9.9 Examples 2-9 75.8 ± 17.3 Examples 2-10 68.6 ± 7.0 Examples 2-11 62.6 ± 5.9 Examples 2-12 65.0 ± 2.9 Examples 2-13 66.4 ± 6.4 Examples 2-14 64.7 ± 3.7 Examples 2-15 75.5 ± 5.9 Examples 2-16 70.4 ± 7.6

<Experimental Example 8> Collagenase activity inhibitory activity

The inhibitory activity of collagenase activity was evaluated according to the mixing ratio of Example 2. For the inhibition of collagenase activity, EnzChek ^® Collagenase Assay Kit - 250-2000 Assays reagent from Invitrogen was used. First, add 1.0 mL of DDW to 1 mg DQ collagen vial to make DQ collagen stock solution (1 mg / mL). Dilute the reaction buffer by adding 18 mL of DDW to 2 mL of 10 x reaction buffer. Next, a collagenase enzyme reagent is prepared. Dilute to working solution with reaction buffer to a final concentration of 0.2 Unit / mL. Prepare the samples to be used at concentrations of 0.01, 0.1, 1, and 10 mg / mL, and prepare the samples in triplicate in a volume of 50 μL on a 96-well plate. DQ collagen 20uL and 30uL of reaction buffer were added to the sample blank and 30μL of working solution was added to the sample. The sample was allowed to stand for 1-2 hours at room temperature in the state of blocking light, and then ELAZA plate reader (SpectraMax M2 & M2e Multi-Mode Microplate Reader, Molecular Devices Corporation, USA).

Collagenase activity inhibition (%) = (est sample-sample blank) / (negative control-blank control) x 100

Collagenase (%) Example 2-1 57.1 ± 4.0 Example 2-2 47.8 ± 4.4 Example 2-3 49.2 ± 2.8 Examples 2-4 37.3 ± 4.4 Example 2-5 103.4 ± 1.6 Examples 2-6 102.8 ± 18.0 Examples 2-7 102.5 ± 13.0 Examples 2-8 102.0 ± 3.4 Examples 2-9 52.9 ± 2.4 Examples 2-10 55.7 ± 4.0 Examples 2-11 58.6 ± 9.0 Examples 2-12 46.3 ± 4.0 Examples 2-13 98.9 ± 4.3 Examples 2-14 95.4 ± 16.2 Examples 2-15 100.2 ± 20.0 Examples 2-16 99.4 ± 60.4

Example 3 Measurement of Activity of Blue Energy Maritime Complex

Through the above Experimental Examples 5 to 8, the mixing method of Example 2-13 confirmed the highest activity, and Example 2-13 was named "Blue Energy Maritime Complex".

Mixing ratio (concentration) Activity (%) oyster Moth Kelp sea hare ABTS Tyrosinase Moisturizng Collagenase Example 2-1 0.5 0.5 0.5 0.5 84.1 46.8 38.9 57.1 Example 2-2 0.5 0.5 0.5 2 77.1 31.5 42.8 47.8 Example 2-3 0.5 0.5 2 0.5 72.6 20.6 66.9 49.2 Examples 2-4 0.5 0.5 2 2 80.4 40.7 54.2 37.3 Example 2-5 0.5 2 0.5 0.5 99.7 84.6 47.9 103.4 Examples 2-6 0.5 2 0.5 2 99.8 73.0 58.8 102.8 Examples 2-7 0.5 2 2 0.5 100.2 85.1 67.6 102.5 Examples 2-8 0.5 2 2 2 99.9 82.1 65.2 102.0 Examples 2-9 2 0.5 0.5 0.5 94.5 35.6 75.8 52.9 Examples 2-10 2 0.5 0.5 2 94.8 36.0 68.6 55.7 Examples 2-11 2 0.5 2 0.5 94.8 31.6 62.6 58.6 Examples 2-12 2 0.5 2 2 91.8 26.1 65.0 46.3 Examples 2-13 2 2 0.5 0.5 99.9 84.2 66.4 98.9 Examples 2-14 2 2 0.5 2 99.7 82.6 64.7 95.4 Examples 2-15 2 2 2 0.5 99.8 83.6 75.5 100.2 Examples 2-16 2 2 2 2 99.7 80.7 70.4 99.4

Experimental Example 9: ABTS radical scavenging ability

The ABTS assay method was modified by the method of Re et al. (1999). That is, 5 ml of 7 mM ABTS and 88 μl of 140 mM potassium persulfate are mixed, and then light is blocked at room temperature for 16 hours to form ABTS cations. This solution was then diluted with PBS to an absorbance value of 0.7 at 734 nm. 190 μl of the prepared diluted solution and 10 μl of the sample were mixed, reacted at room temperature for 6 minutes, and the absorbance was measured at 734 nm. As a solvent control, the radical scavenging ability of the control group was expressed as a percentage.

Dilution factor ABTS radical scavenging activity (%) 200 times 8.5 ± 1.1 100 times 10.8 ± 0.6 50 times 14.0 ± 2.0 20 times 29.3 ± 3.7 10 times 51.2 ± 5.7 5 times 81.8 ± 1.1 Undiluted solution 100.3 ± 3.4

The raw material was diluted to confirm the dilution ratio indicating the activity. As a result, it showed antioxidant activity of 8.5% (0.5% content of cosmetics in cosmetics manufacturing) and about 30% (20% in cosmetics) when diluted 200 times.

Experimental Example 10: DPPH radical scavenging ability

The DPPH assay method is described in Yoshida et al. (1989). 100 μl of deionized water is added to the 96-well plate, and 100 μl of sample diluted with deionized water is added to the test group. Add 100 μl of ethanol into all wells and add 50 μl of a solution of DPPH (0.5 mM 1,1-diphenyl-2-picrylhydrazyl (DPPH) / ethanol). After reacting at room temperature for 30 minutes at 25 ° C, the absorbance was measured at 517 nm. As a solvent control, the radical scavenging ability of the control group was expressed as a percentage.

Dilution factor DPPH radical scavenging activity (%) 200 times 5.2 ± 4.7 100 times 6.0 ± 3.5 50 times 5.6 ± 6.5 20 times 8.6 ± 5.3 10 times 8.5 ± 5.9 5 times 17.2 ± 4.8 Undiluted solution 19.3 ± 5.7

The raw material was diluted to confirm the dilution ratio indicating the activity. As a result, high activity was not confirmed in the DPPH radical scavenging activity.

<Experimental Example 11> Measurement of inhibitory activity of tyrosinase activity

The sample is dissolved in an appropriate solvent and diluted to a suitable concentration range to inhibit tyrosinase activity. Add 220 μl of 0.1 M phosphate buffer (pH 6.5), 20 μl of sample solution and 20 μl of mushroom tyrosinase (2 U / μl) in the test tube in this order. To this solution, 40 μl of a 1.5 mM tyrosine solution was added and reacted at 37 ° C. for 30 minutes. The absorbance is measured at 490 nm using an ELISA reader. Add 0.1 M phosphate buffer (pH 6.5) instead of the sample solution to the blank.

Dilution factor Tyrosinase (%) 20 times 8.7 ± 1.5 10 times 15.1 ± 2.7 5 times 24.9 ± 1.2 Undiluted solution 34.0 ± 4.9

<Experimental Example 11> Measurement of moisturizing effect

To measure the moisturizing effect, 1 ml of the sample was dipped in a cotton swab at 37 ° C in an incubator, and the water holding capacity was measured by measuring the weight per hour. The moisture retention was measured by using distilled water containing no sample as a control. After one hour, the water retention capacity was confirmed little by little, and as the time passed, the water retention capacity was higher than that of the distilled water.

Moisture retention activity (%) 1 hours 2 hours 4 hours DW 69.4 ± 7.0 43.3 ± 7.5 14.3 ± 2.5 Blue energy 75.1 ± 4.1 50.8 ± 4.0 22.7 ± 2.2

<Experimental Example 12> Measurement of collagenase activity inhibitory activity

The inhibitory activity of collagenase activity was measured in the same manner as in Experimental Example 8.

Dilution factor Collagenase (%) 50 times 0.4 ± 1.9 20 times 19.4 ± 5.4 10 times 21.9 ± 5.0 5 times 60.0 ± 7.2 Undiluted solution 96.3 ± 58.3

<Experimental Example 13> Measurement of cytotoxicity against human epidermal cells (HaCaT)

To determine the cytotoxicity against the skin of the sample, toxicity was measured with HaCaT cell, a keratinocyte cell line. The HaCaT cell line cultured in the cell culture flask was cultured in Dulbecco's modified Eagle's medium (LOW GLUCOSE) containing 5% FBS. The HaCaT cell line was plated at a density of 1 × 10 ⁴ / well on a 96-well plate and cultured in a CO ₂ incubator maintained at 37 ° C., 95% humidity and 5% CO ₂ for 24 hours. Sixteen cosmetic raw materials were dissolved in a 1/100 dilution of the cell culture medium and treated in a cell line. Cell culture After 1 day of incubation, cell toxicity was measured. The toxicity of the cells was measured using a MTS reagent in a microplate reader (Molecular Devices, VersaMax ELISA Microplate Reader, USA) at 490 nm. Cell viability was expressed as the absorbance of the sample treated group versus the untreated control.

Dilution factor Cytotoxicity (%) 10 times 104.9 ± 3.8 5 times 106.3 ± 6.0 Twice 111.0 ± 3.4 * Undiluted solution 113.3 ± 6.0 *

* p < 0.05, compared with control group

As shown in Table 16, the 'blue energy maritime complex' of the present invention did not show cytotoxicity.

<Experimental Example 14> Measurement of cell proliferative ability against human epidermal cell (HaCaT)

The ability to proliferate to the skin was confirmed by HaCaT cell, a keratinocyte cell line. That is, the HaCaT cell line cultured in the cell culture flask was cultured in Dulbecco's modified Eagle's medium (LOW GLUCOSE) containing 5% FBS. The HaCaT cell line was plated at a density of 2 × 10 ³ / well on a 96-well plate and cultured in a CO ₂ incubator maintained at 37 ° C., 95% humidity and 5% CO ₂ for 24 hours. The blue energy source was prepared by diluting the blue energy material 10 times from the stock solution to the new medium. The diluted raw material was dissolved in a 1/100 dilution of the cell culture medium and treated with the cell line. The cells were cultured for 1 day, 2 days, and 3 days, and cell proliferation activity was measured. The cell viability was measured at 490 nm using a microplate reader (Molecular Devices, VersaMax ELISA Microplate Reader, USA) using MTS reagent. Cell viability was expressed as the absorbance of the sample treated group versus the untreated control.

Dilution factor Day 1 Day 2 Day 3 10 times 105.6 ± 4.2 * 106.8 ± 3.9 118.8 ± 10.9 * 5 times 103.4 ± 2.3 * 105.5 ± 5.4 123.1 ± 10.3 * Twice 104.3 ± 2.9 * 103.1 ± 4.1 * 125.1 + - 8.7 * Undiluted solution 102.0 + - 4.9 108.7 ± 7.6 * 138.0 + - 10.7 *

* p < 0.05, compared with control group

As shown in Table 17, the 'blue energy maritime complex' of the present invention increased the cell survival rate according to the 1st day, 2nd day and 3rd day.

Production Example of Cosmetic

The blue energy maritime complex according to the present invention can be manufactured into various types of cosmetics according to the purpose. Hereinafter, a method for producing some cosmetics containing the blue energy Maritime complex according to the present invention as an active ingredient is illustrated, but the present invention is not limited thereto.

&Lt; Cosmetic Production Example 1 >

Blue Energy Maritime Complex 10.0 parts by weight

1,3-butylene glycol 1.00 parts by weight

Disodium iodide 0.05 part by weight

Allantoin 0.10 parts by weight

Dipotassium glycyrrhizate 0.05 part by weight

Citric acid 0.01 part by weight

Sodium citrate 0.02 parts by weight

Glycereth-26 1.00 parts by weight

Arbutin 2.00 parts by weight

Hydrogenated castor oil 1.00 parts by weight

ethanol 30.0 parts by weight

Preservative a very small amount

coloring agent a very small amount

Flavoring agent a very small amount

Purified water Balance

<Cosmetic Preparation Example 2> Preparation of nutritional cream

Blue Energy Maritime Complex 10.0 parts by weight

1,3-butylene glycol 7.00 parts by weight

glycerin 1.00 parts by weight

D-Panthenol 0.10 parts by weight

Plant extract 3.20 parts by weight

Magnesium aluminum silicate 0.30 parts by weight

PEG-40 stearate 1.20 parts by weight

Stearic acid 2.00 parts by weight

Polysorbate 60 1.50 parts by weight

Chin type glyceryl stearate 2.00 parts by weight

Sorbitan sesquioleate 1.50 parts by weight

Cetearyl alcohol 3.00 parts by weight

Mineral oil 4.00 parts by weight

Squalane 3.80 parts by weight

Carlyric / capric triglyceride 2.80 parts by weight

vegetable oil 1.80 parts by weight

Dimethicone 0.40 parts by weight

Deposium glycyrrhizae a very small amount

Allantoin a very small amount

Sodium hyaruronate a very small amount

Tocopheryl acetate Suitable amount

Triethanolamine Suitable amount

Preservative Suitable amount

Flavoring agent Suitable amount

Purified water Balance

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is indicated by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (10)

Oyster, kelp, kelp, and a locus.
The oysters and the oysters are ground in water at 80-100 ° C, and then subjected to hydrolysis using protein hydrolysis enzymes.
The cosmetic composition having antioxidative activity, characterized in that the kelp and kelp are dried and then ground and extracted with an organic solvent.
Oyster, kelp, kelp, and a locus.
The oysters and the oysters are ground in water at 80-100 ° C, and then subjected to hydrolysis using protein hydrolysis enzymes.
Wherein the kelp and kelp are dried and then ground and extracted with an organic solvent.
Oyster, kelp, kelp, and a locus.
The oysters and the oysters are ground in water at 80-100 ° C, and then subjected to hydrolysis using protein hydrolysis enzymes.
Wherein the kelp and kelp are dried and then ground and extracted with an organic solvent.
Oyster, kelp, kelp, and a locus.
The oysters and the oysters are ground in water at 80-100 ° C, and then subjected to hydrolysis using protein hydrolysis enzymes.
The cosmetic composition for improving skin wrinkles is characterized in that the kelp and kelp are dried and then ground and extracted with an organic solvent.
5. The method according to any one of claims 1 to 4,
Wherein the organic solvent is selected from one or more selected from the group consisting of methanol, ethanol, butanol, ethyl acetate, acetonitrile, acetone, 1,3-butylene glycol, water and mixtures thereof Composition.
5. The method according to any one of claims 1 to 4,
1.75-2.25 parts by weight, 0.25-0.75 parts by weight seaweed, and 0.25-0.75 parts by weight, based on 2 parts by weight of the oyster.
The method according to claim 6,
Based on 2 parts by weight of the oyster, 2 parts by weight of menthol, 0.5 part by weight of seaweed, and 0.5 part by weight of a part.
5. The method according to any one of claims 1 to 4,
1.75-2.25 parts by weight, 1.75-2.25 parts by weight of sea tangle, and 0.25-0.75 parts by weight, based on 0.5 parts by weight of the oysters.
9. The method of claim 8,
And 2 parts by weight of seaweed, 2 parts by weight of sea tangle, and 0.5 part by weight of the oyster, based on 0.5 part by weight of the oysters.
5. The method according to any one of claims 1 to 4,
The cosmetic composition may be at least one selected from the group consisting of an ointment for external use, a cream, a softening agent, a nutritional lotion, a pack, an essence, a hair tonic, a shampoo, a rinse, a hair conditioner, a hair treatment, a gel, a skin lotion, a skin softener, a skin toner, A lotion, a moisturizing lotion, a nutrition lotion, a massage cream, a nutritional cream, a moisturizing cream, a hand cream, a foundation, a nutrition essence, a sunscreen, a soap, a cleansing foam, a cleansing lotion, a cleansing cream, a body lotion and a body cleanser The cosmetic composition according to any one of claims 1 to 3,