KR101921518B1 - A cosmetic composition comprising fermented spirulina extract for antioxydant and skin winkle improvement - Google Patents

Abstract

Provided is a cosmetic composition for antioxidation and skin winkle reduction comprising a fermented spirulina extract as an effective component. The cosmetic composition according to one aspect of the present invention contains the fermented spirulina extract as an effective component to provide antioxidant and skin wrinkle reduction effects. The cosmetic composition also comprises at least one among the group consisting of a broccoli extract, a kale extract, a mugwort extract, a spinach extract, and a green tea extract.

Description

TECHNICAL FIELD [0001] The present invention relates to a cosmetic composition for antioxidant and wrinkle improvement, which contains ferulic extract of Spirulina as an active ingredient,

The present invention relates to a cosmetic composition containing a ferulic extract of Spirulina as an active ingredient.

Skin is the most basic defensive body that protects the body against irritation in contact with the external environment, and is an important body expressing external beauty.

Since external beauty is not only a standard of judgment that symbolizes youth and health but also a competitiveness that affects business ability by expressing self-confidence, the tendency to manage skin in modern society is increasing.

However, as the age increases, the secretion of various hormones that regulate metabolism decreases, the function of immune cells and the activity of cells decrease, and biosynthesis of immune proteins and biocompatible proteins necessary for the living body is reduced, and ozone layer destruction , The free radicals and free radicals caused by environmental pollution such as physical and chemical stimuli and stress caused by the increase in the normal function of the skin is weakened, aging is promoted, the color is bad and the skin is wrinkled.

In order to prevent such phenomenon and to maintain the skin with a bright tone, physiologically active substances obtained from various known animals, plants and microorganisms were added to cosmetics to prevent skin aging caused by oxidation, thereby trying to improve the skin.

The cosmetics industry is developing a number of products using natural products to reduce skin irritation caused by various chemicals. In addition to low adverse effects on skin, natural materials have recently been increasingly developed as cosmetic raw materials, as consumers have become more receptive to cosmetics using natural materials.

However, a cosmetic product containing a natural-derived extract obtained by an ordinary method has not been sufficiently functionalized, such as skin improving effect, and the skin improving activity is not maintained constantly. Accordingly, there is a continuing need to develop cosmetic products derived from natural products, which have excellent functionalities and excellent biostability and skin improving effect.

Recently, Spirulina is attracting attention as a functional cosmetic material. Spirulina is a blue-green cyanobacterium that grows in the tropics with photosynthesis on the surface of a high-alkaline, salty lake. Spirulina has been noted as a health functional food because it contains a high concentration of proteins and various nutrients.

In particular, it has been officially recognized by the United Nations, the World Health Organization and the Food and Drug Administration (FDA) as a complete food product in terms of safety and nutrition as a health food. It is also a decided food.

Spirulina contains high concentrations of all natural nutrients, especially protein and green leafy vegetables, needed by the organism. It contains five nutrients (protein, carbohydrate, various minerals, fat, various vitamins) necessary for the human body, and even nutrients from 20,000 to 30,000 kinds.

In addition, it has a very high protein content (55 ~ 77%) and various components are composed almost same ratio with the human body, so digestion rate of body is more than 95%, and the digestive function is weakened, the elderly, It is a strong alkaline whole food that does not cause imbalance in nutrition even when used for a long time.

Recently, anticancer effect and various immune function enhancing effects have been reported, and it can be developed as a functional cosmetic material because it contains a large amount of various vitamins. However, there have been no reports on the use of Spirulina as a functional cosmetic material to identify skin physiological activity and skin efficacy.

On the other hand, it is called fermentation that a substance useful for living is produced in the process of decomposing organic matter using an enzyme possessed by a microorganism. Natural products include physiologically active ingredients such as various amino acids, organic acids, and natural antioxidants that are beneficial to skin. It is known that fermentation of natural products through useful microorganisms increases the biologically active component twice before fermentation.

Various cosmetics industry has continued to develop natural cosmetics with enhanced functionality through such fermentation process. Particularly, the fermented natural product increases the skin absorption rate because particles of various functional ingredients are reduced through the fermentation process. In addition, since a new physiologically active ingredient which has not existed can be produced, various functional effects can be exhibited by fermenting a natural product.

Accordingly, the present inventors developed a spirulina extract which can be used as a functional cosmetic material by fermenting Spirulina to develop a cosmetic composition having improved functionality by using Spirulina.

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made keeping in mind the above problems occurring in the prior art, and an object of the present invention is to provide a cosmetic composition comprising a fermented extract of Spirulina.

According to one aspect of the present invention, there is provided a cosmetic composition comprising a ferulic extract of Spirulina as an active ingredient.

In one embodiment, the Spirulina fermented extract may be obtained by inoculating a fermentation strain, Bacillus amyloliquefaciens .

In one embodiment, the cosmetic composition may further include at least one selected from the group consisting of broccoli extract, kale extract, mugwort extract, spinach extract, and green tea extract.

In one embodiment, the cosmetic composition may be for antioxidant and wrinkle improvement.

In one embodiment, the extract may be extracted with one or more solvents selected from the group consisting of purified water, methanol, ethanol, butylene glycol, and propylene glycol.

In one embodiment, the cosmetic composition is selected from the group consisting of a skin, a lotion, a cream, a serum, an emulsion, an essence, a powder, a foundation, a spray, a mask pack, a sheet pack, a sleep pack, Or more.

The cosmetic composition according to one aspect of the present invention contains the spirulina fermented extract as an active ingredient, and thus has excellent antioxidative and wrinkle-reducing effects.

It should be understood that the effects of the present invention are not limited to the effects described above, but include all effects that can be deduced from the description of the invention or the composition of the invention set forth in the claims.

1 is a graph comparing cytotoxic effects of a composition according to an embodiment of the present invention with a control group.
2 is a graph comparing the antioxidant activity of a composition according to an embodiment of the present invention with a control group.
FIG. 3 is a graph comparing the wrinkle-improving effect of a composition according to an embodiment of the present invention with a control group.

As used herein, the terminology used herein is intended to encompass all commonly used generic terms that may be considered while considering the functionality of the present invention, but this may vary depending upon the intent or circumstance of the skilled artisan, the emergence of new technology, and the like. Also, in certain cases, there may be a term selected arbitrarily by the applicant, in which case the meaning thereof will be described in detail in the description of the corresponding invention. Therefore, the term used in the present invention should be defined based on the meaning of the term, not on the name of a simple term, but on the entire contents of the present invention.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Terms such as those defined in commonly used dictionaries are to be interpreted as having a meaning consistent with the contextual meaning of the related art and are to be interpreted as either ideal or overly formal in the sense of the present application Do not.

The numerical range includes numerical values defined in the above range. All numerical limitations of all the maximum numerical values given throughout this specification include all lower numerical limitations as the lower numerical limitations are explicitly stated. All the minimum numerical limitations given throughout this specification include all higher numerical limitations as the higher numerical limitations are explicitly stated. All numerical limitations given throughout this specification will include any better numerical range within a broader numerical range, as narrower numerical limitations are explicitly stated.

Hereinafter, embodiments of the present invention will be described in detail, but it should be apparent that the present invention is not limited by the following examples.

According to one aspect of the present invention, there is provided a cosmetic composition comprising a ferulic extract of Spirulina as an active ingredient.

The cosmetic composition of the present invention contains the ferulic extract of Spirulina as an active ingredient and has excellent antioxidative and anti-wrinkle effects and minimizes skin irritation.

The above-mentioned " comprising as an active ingredient " is intended to mean an effective amount of a composition capable of exhibiting a skin improving effect, such as collagen synthesis associated with a wrinkle-reducing effect, elasticity improvement or skin whitening, skin irritation alleviation, It can mean to do.

The "Spirulina" is the oldest algae on the planet and is a multicellular organism with thin walls. The spirulina is naturally grown in a highly salted alkaline environment and can be mass-produced in artificial ways. The spirulina is spiral in a cyan green color and has a length of 300 to 500 탆 and a width of 8 탆. Approximately 30 species are known and consist of 60 ~ 70% protein, 6 ~ 9% lipid and 15 ~ 20% carbohydrate.

The spirulina includes a variety of known spirulina species, including Spirulina platensis, Spirulina maxima, Spirulina subsalsa , Spirulina major, Spirulina geitleri, Spirulina siamese, Spirulina princeps , Spirulina laxissima , Spirulina curta , Spirulina spirulinoides , Spirulina platensis , Spirulina maxima, Spirulina geitleri and Spirulina siamese .

The spirulina is an antioxidant coloring matter component (chlorophyll, carotenoid, picocyanin), antioxidant enzyme (SOD) and the like including proteins, carbohydrates and water-soluble dietary fibers. Vitamins include beta carotene, vitamin B1, B2, B6, B12, vitamin E, inositol, and folic acid. Calcium, iron, potassium, magnesium, zinc, manganese, selenium and germanium. It contains all the essential amino acids, linolenic acid and gamma linolenic acid are essential fatty acids.

The term " fermentation " as used herein means a process of decomposing an organic substance using an enzyme of the fermentation strain itself. It is known that raw materials after fermentation through fermentation strains are at least two times to several tens of times more effective than before fermentation. Fermented metabolites contain various amino acids, organic acids, and antioxidants that are good for skin, promoting skin metabolism and smooth skin texture. In addition, as the fermentation process proceeds, the particles become smaller and the toxicity such as heavy metals is decomposed, so that the absorption rate is good and the skin troubles and allergy side effects are alleviated.

The spirulina fermented extract may be obtained by sequentially subjecting to sterilization, fermentation, extraction, filtration and concentration.

Specifically, Spirulina is dried, pulverized, mixed with purified water and heat-sterilized at a temperature of 120 to 125 ° C for 10 to 20 minutes. By sterilizing the spirulina, it is possible to prevent intrusion of other contaminants other than the fermentation strain in the subsequent fermentation step. Since the spirulina has a high concentration of various nutrients, it may be contaminated with microorganisms producing toxic substances during fermentation.

Therefore, most spores of microorganisms are destroyed and sterilized at a temperature of 120 ~ 125 ℃ for 10 ~ 20 minutes at which the useful components contained in spirulina are not destroyed. When sterilized at temperatures higher than 125 ° C or longer than 20 minutes, the useful ingredient contained in Spirulina may be destroyed. In addition, if the sterilization time is less than 120 ℃ or the sterilization period is shorter than 10 minutes, harmful microorganisms and spores are not destroyed, and thus there is a high possibility of contamination in the subsequent fermentation step.

The sterilized spirulina is inoculated with a fermentation strain and fermented at a temperature ranging from 30 to 37 ° C for 5 to 10 days. The fermentation strain may be Bacillus amyloliquefaciens .

The above-mentioned Bacillus amyloliquefaciens selected strains having the highest activity among the strains fermenting Spirulina. Bacillus (Bacillus) paper has been selected primarily by fermentation strains fermented spirulina, was named the best zymogen mainly antioxidant and anti-wrinkle effects of those Bacillus amyl Lori kwepe sieon Suga.

Among the selected fermentation strains, Bacillus amyloliquefaciens AMI-0101 was named as an excellent active strain and deposited on September 14, 2017 (KCCM 80155) at the Korean Microorganism Conservation Center.

The fermentation temperature should be within the temperature range of 30 ~ 37 ° C, where the fermentation strain grows best, and the fermentation period should be between 5 and 10 days, which is the strongest growing period of the fermentation strain. If the fermentation temperature is lower or higher than the above temperature range, the growth rate of the fermentation strain may be slow and the fermentation may not proceed properly or may be contaminated with other harmful microorganisms.

The fermented spirulina is dried in a temperature range of 40 to 50 ° C for 12 to 24 hours to obtain a spirulina solid fermented product, and the spirulina solid fermented product is mixed with an extraction solvent and stirred for 5 to 10 days. The extract is filtered and concentrated under reduced pressure and lyophilized to obtain a ferulic extract of Spirulina.

Meanwhile, the cosmetic composition may further include at least one selected from the group consisting of broccoli extract, kale extract, mugwort extract, spinach extract, and green tea extract.

The "broccoli (Brassica oleracea "is a kind of vegetable belonging to Brassica oleracea, a kind of pear. Broccoli is rich in vitamin C and is known to contain a large amount of anticancer substances. When broccoli is cooked and consumed, more than 30mg of vitamin C is known to be released. When broccoli is ingested, the secretion of the immune substance into the digestive system can improve the effectiveness of the anticancer treatment.

In addition, broccoli contains glucosamine and plays a role in generating anticancer substances called sulfolafan. Beta-carotene, which is contained in broccoli, is the pre-production of vitamin A and has antioxidant properties. It eliminates the harmful oxygen that accumulates inside the body and prevents adult diseases such as aging, cancer and heart disease. However, if broccoli is boiled in water for more than 10 minutes, all nutrients will be destroyed.

The " kale ( Brassica There is oleracea . acephala . ) Is a two-year-old or perennial plant of the crucifix, and has a variety of curly kale, spamcale, and flower kale. The kale contains a variety of nutrients because it contains chlorophyll, calcium, phosphorus, fiber, vitamins A, B, C, E and K and is rated as the 'best vegetable' by the World Health Organization (WHO). The kale promotes the blood-forming hematopoiesis, which is good for anemia. It acts on the blood, cleansing the bowel, promoting metabolism, and generating new cells. In addition, the kale enhances the antibody against human contaminants to enhance the hypersensitivity against bacterial infection, stabilizes the cell membrane tissue to prevent aging, prevents dry scalp and prevents hair loss, activates the immune system It is effective in preventing acne.

The " Artemisia " princeps "is a perennial herb that belongs to Asteraceae and is distributed in Korea, Japan, and China and grows in sunny grasses. The mugwort was used for the treatment of bruises due to eczema, wounds, The mugwort is rich in vitamins A and C and is known to be a good food for preventing and treating skin dryness, respiratory diseases, various allergic symptoms, and gastrointestinal diseases. Vitamin A, which is rich in mugwort, helps differentiate epithelial cells and has antioxidant effect. Vitamin C is a nutrient essential for collagen synthesis, which is a protein that constitutes connective tissues. It keeps skin elastic.

In addition, esculin, a physiologically active substance of mugwort, has been reported to have an anticancer activity as a coumarin derivative (Aoife & Richard, 2004), and an activity study on various derivatives (Kostova, 2005) has been conducted. In general, esculin is extracted from the marroni seeds and has the activity of vitamin P, absorbs ultraviolet light and emits fluorescence when irradiated with ultraviolet light of 365 nm. The seeds of Marroni have been used to treat inflammation and vascular disease.

The " Spinacia " oleracea "is the origin of the region in the southwestern part of Asia with the year of the Baekjae or the year of the biennial. The spinach grows up to about 50 cm in height, and leaves and young leaves on the roots are edible as herbs and are used as medicines for constipation by folk remedies. The spinach is abundant in magnesium, potassium, phosphorus, calcium, etc., and contributes to fatigue recovery and improvement of immunity. In particular, potassium releases waste and heavy metals and strengthens skin moisturizing power. Magnesium regulates sebaceous secretion, .

The above-mentioned " Camellia Sinensis " is an evergreen broad-leaved shrub belonging to the genus Chrysanthemum. Especially antioxidant activity, anticancer activity, blood lipid lowering effect in cardiovascular system, and blood circulation promoting action. In particular, Jeju green tea was used in the present invention.

The "extract" is a natural extract using various natural products as an extraction raw material. If the extract is a substance in which a solvent and an extraction raw material are brought into contact with each other under specific conditions to separate the components contained in the raw material from the natural product as an active ingredient contained in the extraction raw material, Regardless of the kind of the component or the component.

For example, it may include extracts of components dissolved in a solvent from natural products by using water or an organic solvent, and specific components of natural products, such as oils obtained by extracting only specific components such as oil. The extract may be dried and pulverized, or may be obtained by various extraction methods known in the art such as a hot water extraction method and an ethanol extraction method.

Wherein the extract is selected from the group consisting of water, an anhydrous or lower alcohol having 1 to 4 carbon atoms, acetone, petroleum ether, ethyl acetate, butyl acetate, trichloromethane, dichloromethane, chloroform, hexane and 1,3-butylene glycol , And can be extracted with ethanol in particular.

The extraction ratio of the active ingredient contained in the raw material may be different depending on the polarity of the solvent. Since the ethanol is excellent in selectivity for extracting a physiologically active substance from a natural raw material, the ethanol extract can realize an optimal skin improving effect .

Since the water and ethanol have different polarities and the effective components to be extracted depend on the polarity, the concentration of ethanol can be appropriately controlled so that an optimal skin improving effect can be realized.

The natural extract is prepared by extracting the extractant with a conventional method such as circulation extraction, pressure extraction or ultrasonic extraction for about 1 to 24 hours with a solvent having a volume of 10 to 20 times the weight of the raw material after it is washed with water, And filtering. In addition, the extract can be obtained in powder form by an additional process such as vacuum distillation or freeze-drying.

The extract may also contain an extract which has been subjected to a conventional purification process. For example, the extract may be subjected to various purification methods such as separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity) Lt; RTI ID = 0.0 > fractions. ≪ / RTI >

The cosmetic composition may be formulated into at least one member selected from the group consisting of a skin, a lotion, a cream, a serum, an emulsion, an essence, a powder, a foundation, a spray, a mask pack, a sheet pack, a sleep pack, have.

The cosmetic composition may be formulated by a conventional method. The contents disclosed in Remington ' s Pharmaceutical Sciences, Mack Publishing Company, Easton PA in the formulation of the external preparation for skin can be referred to, and in the formulation of the cosmetic composition, International cosmetic ingredient dictionary, 6th ed (The cosmetic, Toiletry and Fragrance Association , Inc., Washington, 1995).

Specifically, the cosmetic composition may be prepared into a general emulsified formulation and a solubilized formulation. For example, lotion such as soft lotion or nutrition lotion; Lotion such as facial lotion, body lotion; Nourishing cream, moisture cream, cream such as eye cream; essence; Makeup ointment; spray; Gel; pack; Sunscreen; Makeup base; A foundation such as a liquid type, a solid type or a spray type; powder; Make-up removers such as cleansing creams, cleansing lotions and cleansing oils; Or a cleansing agent such as a cleansing foam, a soap, a body wash, and the like, but is not limited thereto. In addition, the external preparation for skin may be formulated into ointments, patches, gels, creams or sprays, but is not limited thereto.

The cosmetic composition of the present invention may be appropriately blended with other ingredients within the range of not compromising the object of the present invention depending on the kind of the formulation or the purpose of use, in addition to the essential ingredients in each formulation.

The cosmetic composition may contain a conventional acceptable carrier and may appropriately contain, for example, oil, water, a surfactant, a moisturizer, a lower alcohol, a thickener, a chelating agent, a colorant, no.

The acceptable carrier may vary according to the formulation. For example, when formulated into ointments, pastes, creams or gels, there may be used an animal oil, a vegetable oil, a wax, a paraffin, a starch, a tracer, a cellulose derivative, polyethylene glycol, silicon, bentonite, silica, talc, zinc oxide, Extracts may be used.

When the cosmetic composition is formulated with a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder or an extract thereof may be used as a carrier component. In the case of a spray, Propellants such as carbon, propane, butane or dimethyl ether.

When the cosmetic composition is formulated into a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent may be used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl benzoate, 1,3-butyl glycol oil may be used, and in particular, fatty acid esters of cottonseed oil, peanut oil, corn oil, olive oil, castor oil and sesame oil, glycerol aliphatic ester, polyethylene glycol or sorbitan may be used.

When formulated into a suspension, the cosmetic composition may contain, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the cosmetic composition is formulated into soap, it is preferable to use, as a carrier component, an alkali metal salt of a fatty acid, a fatty acid hemiester salt, a fatty acid protein hydrolizate, isethionate, a lanolin derivative, an aliphatic alcohol, a vegetable oil, Can be used.

The cosmetic composition may be a cosmetic or dermatological composition which is generally used in the art depending on the quality or function of the final product, such as a lipid, an organic solvent, a solubilizing agent, a thickening agent, a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, Surfactants, water, ionic or nonionic emulsifiers, fillers, metal ion sequestrants, chelating agents, preservatives, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, And may further contain adjuvants conventionally used in the field of cosmetics or dermatology, such as any other ingredient.

However, the adjuvant and the mixing ratio thereof can be appropriately selected so as not to affect the preferable properties of the cosmetic composition according to the present invention.

The present invention will be further described with reference to the following examples, but it should be apparent that the present invention is not limited by the following examples.

Manufacturing example  One : Spirulina  Fermented extract

Spirulina was collected and then completely dried and pulverized at room temperature.

The Spirulina crushed product and 3-fold volume of purified water were mixed and sterilized at 121 ° C for 15 minutes.

And 10% (w / v) of Bacillus amyloliquefaciens isolated from sterilized Spirulina emeju was inoculated. Spirulina, inoculated with the fermentation strain, was put into a solid fermenter and fermented at a temperature ranging from 30 to 37 ° C for 10 days.

The fermented spirulina was dried at a temperature of 40 ° C for 24 hours to obtain a solid fermented product. The resulting solid fermentation product was immersed in 70% (v / v) ethanol 20 times its volume and stirred for 7 days.

The extract was filtered through 250 mesh and 3 mu m filters, concentrated under reduced pressure at 60 DEG C, and lyophilized to obtain a solid content of the extract.

Manufacturing example  2 : Spirulina  extract

After collecting the spirulina, it was completely dried at room temperature and pulverized to obtain a pulverized product.

The spirulina powder was immersed in 70% (v / v) ethanol of 20 times volume and stirred for 7 days.

The extract was filtered through 250 mesh and 3 mu m filters, concentrated under reduced pressure at 60 DEG C, and lyophilized to obtain a solid content of the extract.

The cell stability, antioxidative effect and wrinkle-reducing effect of the fermentation extract and extract obtained above were verified in the following experimental examples.

Manufacturing example  3: Preparation of natural extract

Broccoli, kale, wormwood, spinach and green tea were washed with water, completely dried at room temperature, and pulverized to obtain 100 g of the pulverized product.

100 g of each pulverized product was subjected to immersion extraction with a 20-fold volume of ethanol as a solvent at a temperature of 50 캜 for 24 hours.

The extract was filtered through a 250 mesh filter and a 3 탆 filter. The remaining ingredients were repeatedly extracted three times in the same manner, concentrated under reduced pressure at 50 ° C, and lyophilized to obtain a solid fraction of the extract.

Comparative Manufacturing Example  One : Spirulina  Fermented extract

A fermented extract was prepared in the same manner as in Preparation Example 1, except that only the fermentation strain was changed to Bacillus subtilis .

Comparative Manufacturing Example  2 : Spirulina  Fermented extract

A fermented extract was prepared in the same manner as in Preparation Example 1, except that only the fermentation strain was changed to Lactobacillus plantarum .

Example  And Comparative Example

In order to verify the stability and functionality of the obtained extracts, examples and comparative examples were set by mixing samples as shown in Table 1 below.

[Content (parts by weight)] division Example Comparative Example One 2 3 4 5 6 7 One 2 3 4 5 6 7 Spirulina
Fermented extract
(Production Example 1) 60 50 50 50 50 50 50 - - - - - - - Spirulina extract
(Production Example 2) - - - - - - - 60 - - - 50 - - Spirulina
Fermented extract
(Comparative Production Example 1) - - - - - - - - 60 - - - 50 - Spirulina
Fermented extract
(Comparative Production Example 2) - - - - - - - - - 60 - - - 50 Broccoli extract
(Production Example 3) - 10 - - - - 2 - - - 12 2 2 2 Kale extract
(Production Example 3) - - 10 - - - 2 - - - 12 2 2 2 Mugwort extract
(Production Example 3) - - - 10 - - 2 - - - 12 2 2 2 Spinach extract
(Production Example 3) - - - - 10 - 2 - - - 12 2 2 2 Green tea extract
(Production Example 3) - - - - - 10 2 - - - 12 2 2 2

Experimental Example  1: Skin safety test

MTT assay experiments were performed on fibroblasts (HDF), melanocytes (B16F10), and keratinocytes (HaCaT) to verify skin safety for the samples of the Examples and Comparative Examples.

Samples of the Examples and Comparative Examples were suspended in purified water at concentrations of 0, 1, 5, 10, 20, 50 and 100 μg / mL, respectively, and cell viability was measured.

Cytotoxicity was performed by modifying the method of Mosmann to measure cell viability using MTT {3- (4,5-dimethylthiazol-2-yl) -2-5-diphenyltetrazolium bromide} reagent.

(HDF), melanocyte-forming cells (B16F10), and keratinocytes (HaCaT) were dispensed in 96-well plates at a concentration of 1 × 10 ⁴ cells / well and cultured at 37 ° C. and 5% CO ₂ for 48 hours Respectively.

After the culture medium was removed and the sample was cultured for 48 hours in a concentration-treated medium, the medium was removed and repeatedly washed with PBS (phosphate buffered saline).

MTT was dissolved in PBS at 5 mg / mL and 50 L was added. The cells were cultured at 37 ° C and 5% CO ₂ for 48 hours. 100 L of DMSO (dimethyl sulfoxide) was added per well, and the mixture was stirred for 10 minutes and absorbance was measured at 540 nm. As a control, Transforming Growth Factor (TGF) 0.01 μg / ml was used (FIG. 1).

As a result of measurement, cytotoxicity was not confirmed at all concentrations of each sample. The results suggest that the sample is not only harmless to the human body but also has excellent human safety.

Experimental Example  2 : DPPH  Evaluation of free radical scavenging activity by measuring method

The extracts of Examples and Comparative Examples were suspended in purified water by concentration to evaluate free radical scavenging activity.

The DPPH assay measures absorbance at 540 nm to the extent that the inhibitor decolorizes the stable radical DPPH (2,2-Diphenyl-1-picrylhydrazyl radical).

Samples of the Examples and Comparative Examples were diluted to a concentration of 15.6, 31.3, 62.5, 125, 250, 500 and 1000 μg / ml. 100 μL of 5 × 10 ^-4 M DPPH (2,2-Diphenyl-1-picrylhydrazyl) was added to 100 μL of each sample concentration, stirred for 1 minute, reacted for 30 minutes in a 25 ° C. incubator, nm absorbance was measured (Fig. 2).

Referring to FIG. 2, at the concentration of 1000 μg / ml, the DPPH radical scavenging activity was improved to 41.9% in the example, compared with Comparative Example 1 in which the DPPH radical scavenging ability was 32.3%.

The higher the DPPH free radical scavenging ability, the better the antioxidative effect. The result suggests that the fermentation significantly increased the antioxidant activity of spirulina.

In particular, the examples using the fermentation strains of Comparative Examples 2 and 3 as Bacillus amyloliquefaciens showed higher free radical scavenging activity at a concentration of 1000 μg / ml (Table 2).

These results suggest that there is a marked difference in the antioxidant activity of fermented Spirulina due to differences in fermentation strains.

In addition, the free radical scavenging ability of Example 7 was higher than that of Comparative Example 4 containing Broccoli extract, Kale extract, Mugwort extract, Spinach extract and Green tea extract (Table 2), and the antioxidant activity of the sample was remarkably improved due to the ferulic extract of Spirulina .

Experimental Example  3: ABTS  Evaluation of free radical scavenging activity by measuring method

The extracts of Examples and Comparative Examples were suspended in purified water to evaluate free radical scavenging activity.

The ABTS method measures the degree of conversion of the antioxidant to a colorless ABTS at 732 nm by dissolving the blue-green radical, ABTS (2,2'-azinodi [3-ethylbenzthiazolne-6-sulfonic acid]).

The samples of the Examples and Comparative Examples were diluted to concentrations of 15.6, 31.3, 62.5, 125, 250, 500 and 1000 μg / ml, and the radical ABTS scavenging activity was measured (FIG.

SC ₅₀ is the concentration at which 50% of the radical ABTS is eliminated. The lower the value of SC _50, the higher the scavenging activity of free radicals. The higher the free radical scavenging activity, the better the antioxidative effect.

Referring to FIG. 2, the SC ₅₀ was reduced to 131.2 μg / ml in the example, compared with the comparative example 1 having the SC ₅₀ of 148 μg / ml, suggesting that the fermentation significantly increased the antioxidant activity of Spirulina.

In particular, the SC ₅₀ of the examples using the fermentation strains of Comparative Examples 2 and 3 as Bacillus amyloliquefaciens was low (Table 2), suggesting that there is a marked difference in the antioxidant activity of fermented Spirulina due to the difference of fermentation strains do.

In addition, referring to Table 2, the free radical scavenging ability of Example 7 was higher than that of Comparative Example 4 containing Broccoli extract, Kale extract, Mugwort extract, Spinach extract and Green tea extract, Suggesting a significant improvement.

division DPPH Scatters
(1000 μg / mL) ABTS Scatters
(SC ₅₀ ) Example 1 41.9% 131.2 μg / ml Example 2 42.9% 128.6 μg / ml Example 3 46.6% 129.4 μg / ml Example 4 44.3% 127.6 μg / ml Example 5 42.5% 127.1 μg / ml Example 6 41.4% 127.4 μg / ml Example 7 51.4% 119.8 μg / ml Comparative Example 1 32.3% 148.0 μg / ml Comparative Example 2 39.4% 140.4 μg / ml Comparative Example 3 37.5% 145.8 μg / ml Comparative Example 4 36.4% 146.6 μg / ml Comparative Example 5 37.6% 138.4 μg / ml Comparative Example 6 39.3% 136.4 μg / ml Comparative Example 7 39.1% 144.2 μg / ml

Experimental Example  4 : Elastase  Evaluation of inhibitory activity

The extracts of Examples and Comparative Examples were suspended in purified water by concentration to evaluate elastase inhibition activity.

Elastin is a protein that maintains skin elasticity in skin connective tissue with collagen. Elastase is an alkaline soluble enzyme that degrades elastin. When it suppresses alestase, elastin degradation is inhibited and skin elasticity can be maintained.

Substrate solution was added to 140 μL of Tris buffer (0.2 M Tris buffer, pH 8.0), 20 μL of each concentration sample, and 20 μL of 5 mM N-Succinyl-Ala-Ala-p-nitroanilide (Sigma, S4760). After mixing with 20 μL of 10 μL / mL of elastase (Sigma, E7885) and reacting at room temperature for 15 minutes, the amount of p-nitroaniline produced at 410 nm was measured (FIG.

Referring to FIG. 3, the inhibitory activity of elastase at the concentration of 1000 μg / ml was improved to 33.9% in the example of the present invention, which was 29.5%.

The higher the inhibitory activity of elastase, the better the wrinkle improving effect. The above results suggest that the effect of improving the wrinkle of Spirulina by fermentation is remarkably improved.

In particular, the example using the fermentation strain of Comparative Example 2 and Comparative Example 3 as Bacillus amyloliquefaciens showed a high inhibitory activity against elastase at a concentration of 1000 μg / ml (Table 3).

These results indicate that the effect of improving the wrinkles of fermented Spirulina due to the difference of the fermentation strains is significantly different, and that the effect of improving the wrinkles is remarkably improved by fermentation of Bacillus amyloliquefaciens.

As shown in Table 3, the inhibitory activity against elastase was higher in Example 7 than in Comparative Example 4 containing Broccoli extract, Kale extract, Mugwort extract, Spinach extract and Green tea extract, Suggesting that the effect is significantly improved.

Experimental Example  5: Procollagen  Synthetic activity evaluation

The extracts of Examples and Comparative Examples were suspended in purified water in concentrations to evaluate the procollagen synthesis activity.

Human fibroblast cells were seeded in 96-well plates at a concentration of 1 × 10 ⁴ cells / well and cultured at 37 ° C and 5% CO ₂ for 24 hours.

The medium was removed and the cells were maintained in the starvation state for 24 hours, treated with the sample, and cultured for 48 hours.

Cell culture medium was collected and the amount of procollagen was measured using Procollagen Type-1 C-Peptide (PIP) Elisa kit (Takara Bio Inc., JAPAN). The control group was not treated with the sample, and the amount of procollagen increased when the sample was added to the control group was measured in%. Transforming growth factor (TGF) 0.01 μg / ml was used as a positive control (FIG. 3).

Referring to FIG. 3, at 100 μg / ml concentration, pro-collagen synthesis was improved to 245.1% in the example, compared to the comparative example of 152.1%. These results indicate that the wrinkle improving effect is increased as the synthesis of procollagen is improved, and the wrinkle improving effect of Spirulina is remarkably improved by fermentation.

As shown in Table 3, the amount of procollagen produced at the concentration of 100 μg / ml was significantly improved in the examples using the fermentation strains of Comparative Examples 2 and 3 as Bacillus amyloliquefaciens.

These results indicate that the effect of improving the wrinkles of fermented Spirulina due to the difference of the fermentation strains is significantly different, and that the effect of improving the wrinkles is remarkably improved by fermentation of Bacillus amyloliquefaciens.

In addition, the activity of synthesizing procollagen of Example 7 was higher than that of Comparative Example 4 containing Broccoli extract, Kale extract, Mugwort extract, Spinach extract and Green tea extract (Table 3) Suggesting a significant improvement.

division Elastase Inhibition
(1000 μg / mL) Procollagen  synthesis
(100 μg / ml) Example 1 33.9% 245.1% Example 2 34.3% 266.6% Example 3 36.5% 241.2% Example 4 34.7% 245.4% Example 5 36.1% 253.5% Example 6 35.5% 265.1% Example 7 38.6% 271.2% Comparative Example 1 29.5% 152.1% Comparative Example 2 30.5% 187.2% Comparative Example 3 29.4% 175.3% Comparative Example 4 29.6% 163.6% Comparative Example 5 24.8% 195.8% Comparative Example 6 30.1% 184.1% Comparative Example 7 30.9% 179.3%

It will be understood by those skilled in the art that the foregoing description of the present invention is for illustrative purposes only and that those of ordinary skill in the art can readily understand that various changes and modifications may be made without departing from the spirit or essential characteristics of the present invention. will be. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive.

For example, each component described in a single form may be distributed and implemented, and components described as being similarly distributed may also be implemented in a combined form. The scope of the present invention is defined in the appended claims And all changes or modifications derived from the meaning and scope of the claims and their equivalents are to be interpreted as being included within the scope of the present invention.

Claims (3)

Spirulina fermented extract; And
Wherein the active ingredient is at least one selected from the group consisting of broccoli extract, kale extract, mugwort extract, spinach extract, green tea extract,
The cosmetic composition for antioxidant and wrinkle improvement obtained by inoculating Bacillus amyloliquefaciens , a fermentation strain, of the fermented extract. The method according to claim 1,
Wherein the extract is extracted with at least one solvent selected from the group consisting of purified water, methanol, ethanol, butylene glycol and propylene glycol. The method according to claim 1,
Wherein the cosmetic composition is at least one formulated cosmetic composition selected from the group consisting of a skin, a lotion, a cream, a serum, an emulsion, an essence, a powder, a foundation, a spray, a mask pack, a sheet pack, a sleep pack, .

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