KR101238708B1 - A method of increasing vitamin c in safflower buds by means of conditioning led light source and a composition containing the safflower buds obtained from the methods - Google Patents

Abstract

PURPOSE: A method for enhancing vitamin C in safflower sprouts and a cosmetic composition containing the safflower sprouts are provided to induce stable photosynthesis and to enhance the vitamin C content. CONSTITUTION: A method for enhancing vitamin C in safflower sprouts under an LED light source condition comprises: a step of germinating safflower seeds for three days and seeding; and a step of irradiating the safflower sprouts with an LED blue light source at a wavelength of 430-460nm. A cosmetic composition for skin whitening and anti-wrinkling contains a safflower sprout extract.

Description

FIELD OF THE INVENTION The present invention relates to a method for increasing the vitamin C content of safflower sprouts by LED light source conditions and a cosmetic composition containing the safflower sprouts, wherein the safflower buds are obtained from the methods}

The present invention relates to a method of increasing the vitamin C content of safflower sprouts by conditioning an LED light source and a composition containing the safflower sprouts obtained by the above method, A method of increasing the vitamin C content of a safflower bud by irradiating a blue light LED to a safflower bud in a plant plant, and a method of increasing the vitamin C content of a safflower bud by using a method of whitening or wrinkle- To a composition for functional health food.

The cosmetics industry has a strong defensive nature, which continues to grow despite the economic downturn. Among functional cosmetics, whitening cosmetics showed the highest growth rate between 2001 and 2006, while functional cosmetics showed the highest share after basic cosmetics.

Recent consumer trends favor functional cosmetics of natural materials rather than chemically synthesized functional cosmetics. This is because the image of natural products is not only environmentally friendly and friendly to consumers, but also their effectiveness and effects have been verified by people for a long time, and they are widely used because of their wide variety of natural materials.

However, it is difficult to obtain the homogeneity of the components and quality according to the cultivation and storage of the natural material, and as a result, the content of the active ingredient may not be uniform, the active ingredient or the pharmacologically active ingredient is unclear, There are disadvantages that are not suitable.

Therefore, a plant plant has been proposed as a systematic agricultural form that can produce plants of a certain quality like industrial products. It is a form of agriculture that combines cutting edge IT technology that adopts the latest technology such as LED, environmental control system and robot automation process.

Safflower is an annual plant belonging to the Asteraceae family. It lives in Korea, China, India, etc., and is cultivated much as red dye and medicinal plants in Korea. In Oriental medicine, it is known that safflower seed has a medicinal effect on bone diseases such as fracture and osteoporosis. It is known that safflower flower has a medicinal effect on menstrual pain.

In connection with safflower, Korean Patent Application No. 10-2003-0047056, entitled "Method of germination of safflower seed and method of manufacturing healthy food using same sprout", discloses an invention using ginseng safflower seed for health food, 10-0670237 'Cosmetic composition containing safflower seed extract and method for extracting cosmetic active ingredient in safflower seed' describes an invention of extracting an effective ingredient of cosmetic ingredient from safflower seed by extraction of hot water and using it as a cosmetic composition.

However, none of the prior art discloses the use of the safflower sprout itself in the cosmetic composition, and furthermore, a method of increasing the vitamin C content of the safflower sprout by using LED blue light when cultivating safflower sprout is further described none. In addition, there is no example of prior art fermenting safflower sprout as a means to increase whitening activity.

On the other hand, existing skin care has been mostly used for skin care using cosmetics. However, recently, as the concept of eating cosmetics has formed a trend, the tendency to improve the body through oral ingestion and to bring out fundamental beauty is getting stronger. As reflected in this, in the present day, health food supplements for the treatment of spots, freckles and whitening are distributed through various drug stores including drugstores, but examples of addition of safflower sprouts with increased vitamin C content to food compositions are also disclosed in the prior art I can not.

1. Korean Patent Application No. 10-2003-0047056 (Publication No. 1020050007032, Publication Date: Jan. 17, 2005) 2. Korean Patent No. 1006702370000 (Publication No. 1020050046114, Publication Date: May 18, 2005)

In the present invention, when a natural product is used in a cosmetic composition or a functional health food composition, the content of the active ingredient contained in the natural product is not constant as the cultivation conditions are changed from year to year, so that it is not suitable for use as a raw material for a cosmetic composition or a functional health food composition To solve the conventional problems.

In addition, the present invention discloses a method for economically mass-producing safflower sprouts which can be harvested only in a small amount at a certain time.

In the present invention, optimization cultivation conditions of safflower sprouts are revealed.

Further, in the present invention, the content of the whitening and / or wrinkle improving active compound is increased to improve the function of the whitening and wrinkle improving cosmetic composition.

In addition, the present invention provides a functional health food composition having an increased vitamin C content and excellent skin cosmetic properties.

According to the present invention, a whitening cosmetic composition comprising safflower sprouts and a functional health food composition for skin cosmetics are provided.

Also, according to the present invention, there is provided a whitening cosmetic composition and a functional health food composition comprising safflower sprouts stably subjected to a photosynthesis process in a plant plant.

According to the present invention, there is also provided a whitening cosmetic composition and a functional health food composition comprising a safflower bud with increased vitamin C content by irradiating LED blue light in a wavelength range of 430 nm to 460 nm in a plant plant.

According to the present invention, there is provided a whitening cosmetic composition comprising a safflower bud having an increased vitamin C content of a safflower bud by irradiating an LED blue light in a wavelength range of 430 nm to 460 nm in a plant plant for 10 hours a day for 7 days, To provide a food composition.

In addition, the present invention provides optimization conditions for the cultivation of safflower sprouts in the cultivation of safflower sprouts in plant factories.

According to the present invention, it is possible to massively cultivate safflower sprouts by optimizing irradiation dose, temperature, humidity, etc. in a plant plant, and thus it is possible to provide an environmentally friendly cosmetic raw material production technology.

In addition, safflower sprouts cultivated in plant factories contain a certain amount of effective ingredients, so that stable quality maintenance and supply of functional cosmetics and functional health food materials has become possible.

In addition, it is possible to increase the vitamin C content by optimizing irradiation and cultivation conditions of LED light sources in a specific wavelength range.

In particular, safflower buds have 50 to 70% of melanin inhibitory activity at a concentration of 50 ug / ml and exhibit a collagen aggregation performance of 20%, which is superior to other natural materials as well as superior to that of safflower seed and safflower powder. Corresponds to the effect of improving wrinkles.

In addition, according to the present invention, a model of green cosmetic without carbon dioxide emission is presented.

FIG. 1 is a graph showing polyphenol content of a safflower extract obtained by different drying methods.
FIG. 2 is a graph showing the content of flavonoids in the safflower bud extract with different drying methods. FIG.
FIG. 3 is a graph showing the SOD-like activity of the safflower bud extract with different drying methods.
FIG. 4 is a graph showing the electron donating ability of the safflower bud extract with different drying methods.
FIG. 5 is a graph showing the results of the hydroxy radical scavenging ability test of the safflower bud extract with different drying methods.
6 is a graph showing the results of melanin synthesis inhibition test according to souring culture conditions.
FIG. 7 is a graph showing the results of collagen synthesis performance test according to safflower growing conditions.
FIG. 8 is a graph showing the results of the test for the inhibition of nitrite synthesis according to the safflower growing conditions.
FIG. 9 is a graph showing the results of the test for inhibiting intracellular stress according to the conditions for cultivation of safflower.
FIG. 10 is a graph showing the results of the test for inhibiting the secretion of ß-hexosaminidase according to the conditions of cultivation of safflower.

The term " safflower sprout " as used herein means a sprout which has been germinated for 3 days after seeding of safflower seed and then sown on a plant and harvested on the 7th day. In the present invention, the country of origin of safflower seed is not particularly limited, but domestic safflower seed is preferably used.

As used herein, the term "plant plant" refers to a closed crop growing system that artificially controls the cultivation environment so as to provide a suitable environment for growing crops to obtain more yields and good quality agricultural products. There are two types of plants: plant plant and closed complete control plant growth plant [refer to Korean Journal of Agricultural Machinery Vol. 26, No. 3, June 2001].

Currently, a plant plant system (hereinafter referred to as " plant plant system ") has been proposed in which a plurality of trays having the same structure as a planting space in which plants are grown are stacked on a coaxial plane to maximize hydroponic cultivation area in a limited space, 10-0512378), and a plant plant (Korean Patent Laid-Open Publication No. 10-2004-0010426) in which light of a wavelength necessary for a plant is irradiated intensively and balancedly by using an LED light source is known.

In the present invention, there is no particular limitation on the structure and the control system of a plant plant so that safflower sprouts can stably perform photosynthesis.

The light source used in the present invention is an LED light source. More specifically, an LED light source in the wavelength range of 430 nm to 460 nm is used in the present invention. The wavelength range is the wavelength range found to be suitable for photosynthesis by Engelmann's experiment. In the present invention, the LED light source is installed in a plant factory in a manner customary in the art.

Flavonoid is a plant secondary metabolite commonly referred to as vitamin P, which is known to have antioxidant activity in vitro and has recently received attention for its preventive effect on cancer and cardiovascular diseases.

Vitamin C is different from conventional sunscreen agents and has ultraviolet light blocking function. Once it enters the skin, it has the effect of being stored, so it can maintain its efficacy for a long time even if it is discontinued. In addition, vitamin C has an effect of inhibiting melanin formation, which is deeply related to pigment deposition, and thus improving pigmentation. Vitamin C is mostly biosynthesized from glucose in animals, whereas man should be supplied from glucose. As described above, when vitamin C is deficient, wrinkles are easily formed, the skin becomes rough, various skin protecting functions are deteriorated, and skin lesions occur. Suitable vitamin C containing compositions for skin protection can be administered directly or transdermally.

The mechanisms by which blue light increases the levels of phlorobonoids and vitamin C have not been elucidated yet, but it has been found that blue light is mainly used for photosynthesis in Engelman experiments, and particularly in the last stage of chlorophyll synthesis, high levels of blue light are required [Journal of Biological & Environment Control, v. 2008]. 17, no. 2, 2008, pp. 116-123].

Fermentation / bioconversion was carried out by selecting a variety of enzymatic activities such as bacillus, aspergillus, and mushroom bacteria, especially those with high water activity, such as glucosidase activity, , That is, a process of making the penetration activity of the skin high by making the polymer low-molecular-weight.

As the fermentation method in the present invention, a fermentation method commonly used in the art, for example, an alcohol fermentation method, may be applied, but is not limited thereto.

In the present invention, alcoholic fermentation of safflower sprouts significantly increased skin penetration activity compared to the control without alcohol fermentation.

The safflower bud of the present invention may be contained in the cosmetic composition or the functional health food composition in an amount of 0.01 to 5% by weight based on 100% by weight of the composition.

The cosmetic composition comprising the safflower bud of the present invention may contain conventional auxiliary agents such as antioxidants, stabilizers, solubilizing agents, vitamins, pigments and flavoring agents, and carriers, which are conventionally used in cosmetic compositions.

The cosmetic composition of the present invention can be prepared into any of the formulations conventionally produced in the art and can be used in the form of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, , Oil, powder foundation, emulsion foundation, wax foundation and spray, but is not limited thereto. More specifically, it can be manufactured in the form of a flexible lotion, a convergent lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

When the formulation of the present invention is a paste, cream or gel, an animal oil, vegetable oil, wax, paraffin, starch, tragacanth, 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 spraying, chlorofluorohydrocarbons, propane / Or a propellant such as dimethyl ether.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component.

When the formulation of the present invention is a suspension, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as polyoxyethylene sorbitol ester, microcrystalline cellulose or the like may be used as a carrier component.

When the formulation of the present invention is a surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate and the like may be used as a carrier component.

The cosmetic composition of the present invention can be applied to face or the like once or several times per day.

The functional health food composition according to the present invention contains 0.01 to 5% by weight of safflower bud based on 100% by weight of the composition. If less than 0.01% by weight of safflower bud is used, the desired effect of the present invention can not be obtained. %, The improvement of the effect is not exhibited. The functional health food composition according to the present invention may further comprise at least one auxiliary ingredient selected from vitamin E, L-cystine, glutathione and selenium-containing yeast or extract thereof, zinc oxide and zinc sulfate.

The functional health food composition according to the present invention can be formulated into a form suitable for oral administration, for example, as soft capsules, hard capsules, granules, tablets and the like. Such a composition is preferably in unit dosage form wherein each dosage form contains a minor amount of the active ingredient along with a supplemental ingredient, wherein a suitable diluent, carrier or other excipient is selected according to the usual formulation methods for the pharmaceutical composition, . For example, the tablets may contain, in addition to the active filler, conventional granulating agents, diluents, binders, disintegrants, lubricants, stabilizers, colorants, shading agents, sweeteners and seasoning agents.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the examples are for illustrative purposes only and should not be construed as limiting the invention thereto.

Example

Materials and Preparation

The safflower used in this study was germinated for 3 days and then harvested 7 days after sowing in a plant.

The first drying method is drying for 24 hours in a 60 ° C dryer and the second drying method is for drying the raw material (RM) at -20 ° C Followed by freeze-drying and vacuum drying at -70 ° C for 12 hours using a vacuum freeze dryer (FD5508, Ilshin Lab Co., Ltd., Korea). The pretreated safflower sprouts were pulverized and used as analytical samples. Analytical solvents and the like were HPLC grade solvents (JTbaker) having a purity of 99.8% or more, and reagents of grade 1 or higher were used for the other reagents.

Experimental Example  One: Safflower  Analysis of general components and physicochemical properties

The general components of the samples were measured according to the AOAC method. That is, moisture was measured by atmospheric pressure heating drying, crude protein was measured by micro-Kjeldahl method, crude fat was measured by Soxhlet method, and ash was measured by direct filtration method.

Hereinafter, in the present specification, white light raw materials (WRM) refers to samples irradiated with white light, BRM (blue lighting raw materials) refers to samples irradiated with blue light, RRM (red lighting raw materials) refers to samples irradiated with red light, raw materials) refers to control samples. In addition, WHD (white lighting hot air drying) is a method of spraying safflower buds dried in high temperature air after being irradiated with white light, safflower buds dried in high temperature air after blue light irradiation in BHD (blue lighting hot air drying) drying refers to safflower buds dried in hot air after red light irradiation, and control hot air drying (CHD) controls safflower buds dried in hot air. In addition, white lighting freeze drying (WFD) is a lyophilized sprout which is lyophilized after being irradiated with white light. BFD (blue lighting freeze drying) is a lyophilized sprout which is lyophilized after blue light irradiation. Red lighting freeze drying Controlled freeze drying (CFD) refers to a control lyophilized safflower bud.

(1) General composition analysis

The results of analyzing moisture, crude ash, crude fat and crude protein as general components of safflower sprouts are shown in Table 1 below.

Moisture content was slightly higher in the safflower sprout irradiated with red light depending on the drying conditions. The crude ash content was slightly higher in the safflower buds of blue light than in the safflower bud samples. The crude fat content was somewhat higher in the safflower buds lyophilized after the red light irradiation. The crude protein content of the safflower buds dried at 60 ℃ did not show any significant difference according to the absorption, but that of vacuum freeze dried safflower buds was higher in the safflower buds exposed to red light.

(2) Chlorophyll content

Chlorophyll a stands for cyan, and chlorophyll b stands for yellowish green. The ratio is 3: 1. In addition to the core of chlorophyll is Mg and the center of hemoglobin is Fe, the two substances have the same phorphyrin structure, which means that when the animal is digested by ingesting chlorophyll, the Fe and Mg present in the small villus are replaced Has been reported to be involved in hematopoiesis-producing hemoglobin.

To 3 g of safflower sprouts, 100 ml of 85% acetone was added, and the mixture was pulverized. The mixture was centrifuged at 3,000 rpm for 5 minutes, and then 85% acetate was added to the residue to extract three times. The supernatant was collected and 500 ml was added. Take 25 ml of this solution, add 50 ml of ether and distilled water 25 ml, shake for 1 minute, and take up the ether layer three times. Add a small amount of sodium sulfate to remove water, then use 100 ml of ether to dissolve the absorbance at 660 nm and 642 nm The total chlorophyll, chlorophyll-a and chlorophyll-b contents were measured. The results are shown in Table 2 below.

     Compared with the control group, the content of chlorophyll a showed a higher tendency than that of chlorophyll b, and the chlorophyll content was higher in the safflower sprout with blue light or red light.

(3) Chromaticity

The chromaticity of the safflower buds was measured using a colorimeter (Model CR-200, Minolta Co., Japan). The Hunter scale was used to measure L (Lightness) a (redness), and b (yellowness: yellowness), respectively. A white plate (Y = 94.2, x = -0.3131, y = 0.3201) was used as a standard color plate.

The results of measuring the color of the safflower buds are shown in Table 3 below.

The L value indicating the degree of brightness was measured high in red lighted sunflower safflower. This is attributed to the fact that the freeze - dried safflower sprout, which is free from the influence of temperature, has less browning than the safflower sprout dried by other drying methods.

The a value of redness was the highest in safflower bud samples and lowest in dried safflower at 60 ℃. The value of b indicating yellowness was high in vacuum freeze - dried safflower and there was no significant difference according to absorbance.

(4) Vitamin C content

Dried safflower sprouts were extracted using a sonificator (JAC 4020, Jinwoo, Korea). 2 ~ 3 g of dried safflower sprouts were extracted with 40 ~ 60 ml of 25% EtOH and the sonificator was fixed at 40 ℃. The whole process was carried out over 3 repetitions for 1 hour.

Each extract was filtered with 0.2 ㎛ membrane filter and analyzed by HPLC. The analysis conditions are shown in Table 4. The standard curve was calculated by preparing a standard curve using L (+) - ascorbic acid as a standard reagent.

The results of measuring the vitamin C content are shown in Table 5 below. Vitamin C content was higher in the control group and sunlighted safflower than in the blue light. No vitamin C was detected in the safflower with different drying methods.

(5) Phenolic compound content

The extracts of the safflower sprouts which differed in drying method were extracted with 25% ethanol, and the concentration of 1 mg / mL was measured and the total phenol content was measured. The total phenol content of the safflower sprout was significantly higher than that of the freeze - dried safflower.

(6) Total flavonoid content

The results of analysis of the total flavonoid content of safflower buds with different drying methods are shown in FIG. Blue light showed the highest fluoronoid content in the safflower bud.

2. Antioxidant effect of safflower sprouts in different drying methods

(1) superoxide dismutase (SOD) -like activity

Aerobic organisms produce superoxide radicals through the use of oxygen during respiration metabolism, which is oxidized in association with organisms in living organisms. Oxides cause oxidative damage in living organisms, leading to various diseases in vivo. Therefore, It has a complex antioxidant system by mechanism and non-enzymatic mechanism. It has been reported that polyphenol components such as catechins in photosynthetic metabolism occurring in chlorophyll of plant leaves are closely related to antioxidant ability and phenolic compounds have been reported to have SOD-like activity.

SOD-like activity was measured by adding 3 ml of tris-HCl buffer (pH 8.5) and 0.2 ml of pyrogallol to 0.2 ml of each extract, incubating at 25 ° C for 10 minutes, stopping the reaction with 1N HCl, Ray spectrophotometer.

 A: Absorbance of the sample added group

 B: Absorbance of the sample-free group

The SOD-like activity of safflower sprouts with different drying methods is shown in Fig. According to the present invention, the SOD-like activity of the safflower buds freeze-dried in vacuo was higher than that of the safflower buds dried at 60 ° C.

(2) Electron donating ability (EDA)

The antioxidative activity of each sample was measured by measuring the electron donating ability (EDA) by the DPPH free radical scavenging method.

3 mL of 0.5 mL DPPH (1,1-diphenyl-2-picryl hydazyl) reagent was added to each sample, and the mixture was allowed to stand at room temperature for 30 minutes. A UV-visible light spectrophotometer (Phanrmaca biotech Ultraspec 3000 Engalnad) Lt; / RTI > at 517 nm.

A: Absorbance of the sample added group

B: Absorbance of the sample without added sample

The results of measuring electron donating ability of safflower sprouts according to the drying method are shown in FIG. The results of the search for antioxidant plants in Korean medicinal and plant resources show similar results to that of plant resources with less than 20% of plant resources except for grape seeds.

According to FIG. 4, the red light showed significantly higher electron donating ability in the safflower-dried safflower buds.

(3) Hydroxy radical scavenging activity

The reaction mixture was mixed with FeSO ₄ / EDTA solution, 2-deoxyribose, each extract, phosphate-buffer solution and H ₂ O ₂ for 2 hours. TCA (trichloro acetic acid) solution and TBA (thiobarbituric acid) After rapid cooling, the absorbance at 532 nm was measured to compare the antioxidant activity.

A: Absorbance of the sample added group

B: Absorbance of the sample-free group

The results of measuring the hydroxy radical scavenging ability of safflower sprouts according to the drying method are shown in FIG. As a result of this experiment, it was interpreted that there was no influence by the absorption, and there was no significant difference according to the drying method.

3. Assessment of cell-based efficacy of safflower bud

Effects of water, water absorber, and light condition on the cosmetic effect of safflower sprouts were studied in the plant. This study focused on collagen synthesis performance, anti - inflammation, whitening, antioxidant ability and anti - allergy.

Collagen aggregation performance was measured by proCollagen type 1 peptide method, anti - inflammatory effect was measured by Nitrite assay, whitening effect was measured by melanin content, antioxidant effect was measured by Intracellular ROS detection method , Antiallergic effect was measured by β-hexosaminidase release method.

The used samples were divided into primary to tertiary cultivars. The conditions for the changes are shown in Table 6 below.

Conditions for cultivation of safflower Light condition Water condition Primary 1. Control (Domestic)
2. China (Made in China) - Round three-wave bulb
3. China White - Three-wavelength light bulb + LED (daylight color)
4. China Yellow - Three-wavelength bulb + incandescent lamp 1. Yellow soil ball badge
2. Use common water Secondary 1. White - Three-wavelength light bulb
2. Red - Three-wavelength light bulb + LED (red)
3. Blue - Three-wavelength light bulb + LED (blue)
4. Red + Blue - Three-wavelength bulb + LED (red + blue) 1. Water absorption ball medium (hydrogel)
2. Using ionized water
3. Use Korean domestic safflower seeds Third 1. White - Three-wavelength light bulb
2. Red - Three-wavelength light bulb + LED (red)
3. Blue - Three-wavelength light bulb + LED (blue) 1. Water absorption ball medium (hydrogel)
2. Use common water
3. Use Korean domestic safflower seeds Fourth 1. White - Three-wavelength light bulb 1. Domestic use of safflower seed
2. Spreading baskets on water (long roots without cutting off roots)
3. Use common water

(1) melanin synthesis inhibitory ability

Melanocytes are the differentiated cells in the basal layer of the skin epithelium, which have a special organs called melanocytes that synthesize melanin. Melanin is known to play an important role not only in protecting skin from ultraviolet rays, but also as a scavenger for cytotoxic radicals, as well as pigmentation such as stains and freckles. At present, in order to evaluate the efficacy of whitening raw materials, in vitro tyrosinase activity inhibition test is most commonly used, and intracellular melanin formation inhibition test is most commonly used.

In this experiment, the efficacy of the extract as a whitening ingredient was evaluated by inhibiting melanin synthesis in B16-F1 cells.

murine melanoma (B-16 F1) cells a suitable amount of FBS (fetal bovine serum) containing a after a 6-well plate in DMEM medium inoculated with 1 × 10 ⁵ pieces per well 5% CO _2, the cells are well under 37 ℃ And cultured until it was attached to the bottom of about 80% or more. After removing the culture medium, and samples were incubated under the concentration time constant 5% CO _2, 37 ℃ was replaced with the diluted medium to a suitable. Cells from which the medium was removed were washed with PBS (phosphatized buffer saline), and treated with trypsin to recover the cells. The recovered cells were counted using a hematocytometer, centrifuged at 5,000 to 10,000 rpm for 10 minutes, and the supernatant was removed to obtain a pellet. The cell pellet was dried at 60 ° C, and 100 μl of 1 M sodium hydroxide solution containing 10% DMSO or an appropriate amount of cell lysis buffer was added to obtain intracellular melanin in a 60 ° C thermostat. The absorbance at 405 nm was measured with a microplate reader to determine the amount of melanin per cell number or the amount of melanin per constant protein.

The results are shown in FIG. 6, which shows that the inhibitory activity is 50 to 70% at a concentration of 50 ug / ml.

(2) Collagen aggregation performance

One of the causes of skin wrinkles is the lack of collagen in the skin. Collagen is the main protein that constitutes the skin dermis and plays a role in keeping skin structure and elasticity. It is known that collagen shows a decrease in production with age and is increased in degradation to induce depression of the dermal layer of the skin to produce wrinkles of the skin. Fibroblasts present in the dermis are responsible for the synthesis of collagen, a major protein in the skin. Therefore, by measuring the degree of increase of collagen production using fibroblasts, the collagen synthesis performance of the extract can be improved and the wrinkle improvement effect can be evaluated.

Dermal fibroblasts (HDFn) were inoculated into 48-well plates at a concentration of 5 × 10 ⁴ cells / well in 0.3 ml of culture medium and cultured at 37 ° C and 5% CO ₂ . After 24 hours, the extracts were cultured for 24 hours in a fresh medium containing the concentrations of the extracts. After culturing for 24 hours, the culture broth was collected and the amount of C-terminus of the collagen precursor was measured using an enzymes-linked immunoassay kit (Takara). In addition, a collagen solution (640 ng / ml) was used to obtain a standard curve between 0 ng and 640 ng of collagen concentration, and the amount of collagen in the medium was calculated.

The results are shown in FIG. 7. According to the results, the ability to synthesize collagen increased by about 20% in the samples with blue light.

(3) Nitric Oxide

Various inflammatory mediators are involved in the pathogenesis of inflammation, and the causes of the disease vary. Treatment of macrophages with LPS (lipopolysachride), known as endotoxin, increases NO biosynthesis by inducible NOS (iNOS) expression and induces an inflammatory response. NO is a pathological secondary signaling substance secreted by many cells, regulates the activity of COX and acts synergistically to the inflammatory response. By measuring NO produced by LPS induced by NOS in Raw 264.7 cells, the inhibitory effect of the extract on inflammation can be determined.

Mouse macrophage cell RAW 264.7 cells were treated with LPS, an inflammation inducing substance, to evaluate whether they inhibited inflammation. Cells were seeded at 2 × 10 ⁵ cells per well in a 96-well plate and cultured in a 5% CO ₂ incubator at 37 ° C for 24 hours. Overnight and then replaced with fresh medium. LPS (1 ug / ml) and extracts were added at a concentration ranging from 1 ug / ml to 100 ug / ml and cultured for 24 hours. After incubation, the supernatant was taken and centrifuged at 13,000 rpm for 3 minutes to collect supernatant, reacted with Griess reagent reagent 1: 1, and absorbance was measured at 570 nm.

The results are shown in Fig.

(4) Antioxidant activity test

Energy production in cells involves the production of reactive oxygen species (ROS), a byproduct of mitochondrial respiration, which causes damage to proteins, fats and nucleic acids.

In order to protect the cell itself from such damage, the defense system works in the cell. However, when the balance between the production of free radicals and the antioxidant system is broken, oxidative stress acts on the cell, which is a major cause of various diseases and cancers of the body including aging. do.

Therefore, the antioxidant ability of the extract is evaluated by measuring the amount of active oxygen formed in the cells for developing an antioxidant using a skin-friendly and stable plant as a raw material. DCFH-DA is a nonpolar molecule that can easily enter the cell and is separated by the intracellular esterase into DCFH, which is a nonpolar polar molecule. The DCFH-DA is a typical evaluation method using a fluorimetric assay using 2 ', 7`-dichlorofluorescin-diacetate. After oxidation by reactive oxygen species in the cells, they are converted to fluorescent DCF (2`, 7`-dichlorofluorescin), which is measured by a fluorometer.

The dermal fibroblasts (HDFn) were suspended in DMEM (containing 10% serum, 1% antibiotic), cultured at 37 ° C in a 5% CO ₂ incubator, and cells growing in the larval stage were mainly used for the experiment. Wells were inoculated in 96 wells at a rate of about 1 x 10 ⁵ cells per well. The sample extracts were then treated with various doses and incubated for 20 min at 37 ° C in a 5% CO ₂ incubator. Then, 1 mM of H ₂ O _{2 as an} oxidative stress inducer is added, and 20 μM of DCFH-DA (stock 20 mM) is added thereto, followed by measurement with a spectrofluorophotometer (excitation 485 nm, emission 535 nm). The reaction was observed at 37 DEG C for 90 minutes. The inhibition rate of reactive oxygen species was determined by comparing the number of reactive oxygen species to the total number of active oxygen species in the control group. .

The results are shown in Fig.

Active oxygen species inhibition rate (%) = (control OD-sample treatment group OD) / control OD x 100

(5) Anti-allergy test

Atopic dermatitis is characterized by elevated levels of serum eosinophils and serum IgE, which is a typical indicator of atopic dermatitis. The initial response of atopic dermatitis is due to antigenic stimulation of antibody IgE, When it binds to the receptor, it re-enters the antigen and binds to the antibody that binds to the receptor, which rapidly transfers the signal into the mast cell to secrete chemical mediators (cytokines, histamine, leukotrienes) that cause allergies. Histamine and other mediators increase the vascular permeability, dilate blood vessels, smooth muscle contractions, stimulate the gland function, and the associated inflammatory responses are secreted by mast cells, eosinophils, and T cells It is important that eosinophils migrate to inflammatory sites by various cytokines such as interleukins. It acts as an important mediator with histamine, which has β-hexosaminidase, and the release of β-hexosaminidase from cells is also used as an indicator of immune cell degranulation.

RAB-2H3 cells were suspended in DMEM (Dulbecco's modified Eagle's medium) containing 10% FBS, and then treated with 2 × 10 ⁵ cells per well in a 24-well plate The cells were sensitized with 100 ng / ml mouse monoclonal IgE per well and cultured in a 5% CO ₂ incubator for 24 hours. Cells were rinsed with 500 μl of Tyrode buffer, treated with mixed extracts at various concentrations, incubated at 37 ° C in a 5% CO ₂ incubator for 15 min, and treated with DNP-BSA 100 ng / ml for one hour. The supernatant was treated with 50 μl of substrate (1 mM p-nitrophenyl-N-acetyl-ß-D-glucosaminide), incubated at 37 ° C in a 5% CO ₂ incubator for one hour and absorbance was measured at 405 nm using an ELISA reader.

The results are shown in FIG. 10 and it can be seen that anti-allergic activity showed 40-50% inhibition activity at a concentration of 100 ug / ml, which is the highest concentration of the experiment.

4. Fermentation / bioconversion effect experiment

By fermenting / bioconversion the safflower buds, the glycosylated flavonoids contained in the safflower buds became aglycone-type flavonoids. As a result, the whitening activity was improved by 50% or more.

5. Comparison with other light sources

When blue light was irradiated with safflower sprouts, it was confirmed that the melanin inhibitory activity was increased by 50 to 70% as compared with the case of using white light, and the whitening activity was superior to that of red light (see FIG. 6).

6. Comparison with other natural products

The melanin inhibitory activity and the collagen synthesis performance of other natural products were measured to confirm the whitening effect and the wrinkle improving effect. As a result, it was found that the safflower sprout according to the present invention had remarkably excellent whitening efficacy and wrinkle improving effect.

Formulation example  One: Cosmetics  Preparation of composition

Cosmetic compositions were prepared according to methods customary in the art using the following ingredients:

Safflower bud 3.0 weight%

Butylene glycol 2.0 wt%

Propylene glycol 6.0 wt%

0.5% by weight of carboxyvinyl polymer

Wax 2.0 wt%

Vaseline 7.0 wt%

Polysorbate 60 2.0 wt%

2.5% by weight of sorbitan sesquioleate

Squalane 3.0 wt%

Liquid paraffin 10.0 wt%

0.5% by weight triethanolamine

0.1% by weight of tocopheryl acetate

Spices, preservatives

Purified water balance

Claims (5)

Wherein the amount of polyphenol, flavonoid, and vitamin C is increased at the same time, characterized in that the plant is germinated for 3 days and sowed for 7 days to irradiate the safflower shoot with LED blue light having a wavelength ranging from 430 to 460 nm. delete A cosmetic composition for whitening and wrinkle improvement, which comprises an extract of safflower sprouts grown by the method according to claim 1. A functional health food composition for skin care comprising an extract of safflower sprouts grown by the method according to claim 1. delete

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