KR102380115B1 - Compositions containing safflower seed extract - Google Patents

Abstract

The present invention relates to a composition comprising safflower seed ultrasonic extract as an active ingredient. there is.

Description

A composition containing safflower seed ultrasonic extract as an active ingredient for antioxidant, UV protection and blue light blocking {Compositions containing safflower seed extract}

The present invention relates to a cosmetic composition, a pharmaceutical composition, and a health functional food containing the ultrasonic extract of safflower seed as an active ingredient.

The skin acts as a barrier in the outermost part of the body, protecting various organs and muscles in the body from the external environment. As the heaviest organ in the human body, it also has thermal insulation and temperature regulation functions. It also acts as a sensory organ to various stimuli such as temperature, pressure, and pain, and is also involved in the storage of lipids and water, and the synthesis of vitamin D.

Skin cancer is a generic term for all malignant tumors formed in skin tissue. Skin cancer is mainly caused by external environmental stimuli such as ultraviolet rays or genetic factors. It is known that a family history of several genes, such as the KIT gene, influences skin cancer. However, the factors known to have the greatest influence above all are external stimuli such as ultraviolet rays, the fact that outdoor activities have increased due to the introduction of Western-style lifestyles, the use of tanning and tanning equipment has increased, and the amount of transmitted ultraviolet rays due to the weakening of the ozone layer The incidence of skin cancer is on the rise.

Exposure to a large amount of UV light may cause the formation of a thymidine dimer or double-strand breakage in the DNA inside the cell. If damaged DNA cannot be repaired properly, proto-oncogenes can be turned into oncogenes, and tumor suppressor genes can become dysfunctional. In this case, the division-regulating mechanism inside the skin cells does not work normally, so proliferation is not limited, and normal cells are eventually transformed into tumor cells. Cancerous tissues are supplied with nutrients by attracting peripheral blood vessels through hormone secretion, etc., while cancerous cells themselves are metastasized to other parts of the body through lymph nodes and blood vessels.

A skin cancer that begins to develop in the skin is classified as a 'primary' skin cancer, and a case that metastasizes and settles in the skin after starting a tumor in another organ is classified as a 'metastatic' skin cancer. means skin cancer. Although it is a fairly common cancer among people of Caucasus descent, it has not been actively studied because the fatality rate is generally low compared to cancers occurring in internal organs.

Skin cancer accounts for 2.0% of all cancers in Korea based on 2012 statistics. Diagnosis can be made not only through examination devices such as CT and ultrasound, but also through visual or biopsy. Skin cancer is classified into squamous cell carcinoma, basal cell carcinoma, melanoma, and the like, depending on which cell among the cells constituting the skin tissue is derived. Squamous cell carcinoma, which is a malignant tumor derived from keratinocytes of the epidermis, and basal cell carcinoma, which is a malignant tumor of cells in the basal layer or hair follicle, which is the lowest layer of the epidermis, are quite common skin cancers in Korea, but have a relatively low fatality rate. low. On the other hand, malignant melanoma (malignant melanoma) is a cancer derived from melanocytes that determine the color of the skin, and is a dangerous cancer with the highest malignancy and low survival rate due to good invasion and metastasis.

Blue light is a blue light with high energy in the visible ray region with a wavelength of 380 ~ 500 nm. It is widely emitted from displays such as smartphones and LED lighting devices. In particular, blue light damages mitochondrial DNA in skin cells and generates reactive oxygen species, causing cell dysfunction, cellular aging and tumor development, and inducing melanin production to become the main cause of skin pigmentation or blemishes. . Until now, the development of methods and compositions for protecting the skin from blue light is still insufficient, and the cosmetic industry is developing a number of products using natural substances to replace chemical substances in order to reduce skin irritation with a focus on UV protection. am.

Accordingly, there is a demand for the development of natural substances that can block ultraviolet and blue light and further prevent, improve, or treat skin cancer.

Republic of Korea Patent No. 2141575 Republic of Korea Patent Publication No. 2020-0089040

It is an object of the present invention to provide a cosmetic composition capable of blocking antioxidants, UV rays and blue light by containing the ultrasonic extract of safflower seeds as an active ingredient.

In addition, another object of the present invention is to provide a health functional food containing safflower seed ultrasonic extract as an active ingredient, which can block antioxidants, UV rays and blue light.

In addition, another object of the present invention is to provide a pharmaceutical composition that can prevent or treat skin cancer by containing the ultrasonic extract of safflower seeds as an active ingredient.

The cosmetic composition for antioxidant, UV protection and blue light blocking of the present invention for achieving the above object may include safflower seed ultrasonic extract as an active ingredient.

The safflower seed ultrasonic extract may be extracted under water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

The mixed solvent may be an aqueous solution of 20 to 99.5% by volume of methanol, ethanol, butanol or propanol.

The safflower seed ultrasonic extract may be processed for 10 to 60 minutes with an ultrasonicator of 50 to 700 W power.

The extraction temperature during the ultrasonic treatment may be 20 to 85 ℃.

In addition, the health functional food for antioxidant, UV protection and blue light blocking of the present invention for achieving the above other object may include safflower seed ultrasonic extract as an active ingredient.

In addition, the method for preparing the ultrasonic extract of safflower seeds for antioxidant using the reaction surface analysis method of the present invention for achieving the above another object is (A) safflower seeds with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. Experimental values for total polyphenols in the supernatant obtained by centrifugation after extraction with an extraction solvent, an extraction temperature of 20 to 85 ° C, an extraction time of 10 to 60 minutes, a frequency of 20 to 50 kHz and an ultrasonicator of 50 to 700 W power obtaining; (B) deriving a quadratic regression model represented by the following [Equation 1] using the experimental value of step (A); (C) verifying reliability by performing analysis of variance (ANOVA) on the regression model derived in step (B), and obtaining a response surface curve for the extraction conditions; (D) obtaining an experimental value for the total flavonoid content of the supernatant obtained in step (A); (E) deriving a quadratic regression model represented by the following [Equation 2] using the experimental value of step (D); (F) verifying reliability by performing analysis of variance (ANOVA) on the quadratic regression model derived in step (E), and obtaining a response surface curve for the extraction conditions; (G) obtaining an experimental value for the DPPH radical scavenging ability of the supernatant obtained in step (A); (H) deriving a quadratic regression model represented by the following [Equation 3] using the experimental values of the step (G); (I) verifying reliability by performing analysis of variance (ANOVA) on the quadratic regression model derived in step (H), and obtaining a response surface curve for the extraction conditions; and (J) predicting common optimization conditions for total polyphenol content, total flavonoid content, and DPPH radical scavenging ability by superimposing the response surface curves obtained in steps (C), (F) and (I) may include;

[Equation 1]

Y _TPC =-4.81776-0.015551X ₁ +0.28967X ₂ +0.21412X ₃ +6.72730E-005X ₁ X ₂ +6.01865E-004X ₁ X ₃ -1.00677E-003X ₂ X ₃ -1.23645E-004X ₁ ² - 2.01372 E-0.03X ₂ ² -1.42821E-003X ₃ ²

[Equation 2]

Y _TFC =-0.37372+9.55406E-003X ₁ +9.20547E-003X ₂ +2.05361E-003X ₃ -9.72712E-005X ₁ X ₂ -5.20451E-005X ₁ X ₃ +5.37565E-005X ₂ X ₃ -1.77861E -005X ₁ ² -4.02178E-005X ₂ ² -1.79762E-005X ₃ ²

[Equation 3]

Y _DPPH =-16.21374-0.19557X ₁ +0.64596X ₂ +1.96052X ₃ +5.50661E-004X ₁ X ₂ +2.93686E-003X ₁ X ₃ -3.62518E-003X ₂ X ₃ +4.25974E-004X ₁ ² -2.87540 E-003X ₂ ² -0.013343X ₃ ²

In Equations 1 to 3, Y _TPC is the predicted value of the total polyphenol content (mg GAE / g DM), Y _TFC is the predicted value of the total flavonoid content (mg QE / g DM), Y _DPPH is the predicted value of the DPPH radical scavenging ability ( %), X ₁ is the extraction time, X ₂ is the extraction temperature, and X ₃ is the concentration of the extraction solvent.

Extraction conditions according to common optimization conditions for the DPPH radical scavenging ability, total polyphenol content, and total flavonoid content may be an extraction time of 35.38 minutes, an extraction temperature of 79.38° C., and an ethanol concentration of 80%.

In addition, the pharmaceutical composition capable of preventing or treating skin cancer of the present invention for achieving the above another object may include safflower seed ultrasonic extract as an active ingredient.

The skin cancer may be melanoma.

The composition for antioxidant, UV-blocking, and blue-light blocking, comprising the ultrasonic extract of safflower seed of the present invention as an active ingredient, has excellent antioxidant effects and is effective in blocking UV and blue light as well as improving, preventing or treating skin cancer. It can be used as a health functional food or pharmaceutical composition.

1 is a univariate curve showing the conditions affecting the total polyphenol content (TPC) when using the ultrasonic extract of safflower seed prepared according to an embodiment of the present invention.
2 is a three-dimensional response surface curve of the total polyphenol content (TPC) according to the ultrasonic extraction conditions of safflower seeds prepared according to an embodiment of the present invention.
3 is a univariate curve showing the conditions affecting the total flavonoid content (TFC) when using the ultrasonic extract of safflower seed prepared according to an embodiment of the present invention.
4 is a three-dimensional response surface curve of the total flavonoid content (TFC) according to the ultrasonic extraction conditions of safflower seeds prepared according to an embodiment of the present invention.
5 is a univariate curve showing the conditions affecting the DPPH radical inhibitory activity when using the ultrasonic extract of safflower seed prepared according to an embodiment of the present invention.
6 is a three-dimensional response surface curve of DPPH radical inhibitory activity according to ultrasonic extraction conditions of safflower seeds prepared according to an embodiment of the present invention.

The present invention relates to a cosmetic composition, a pharmaceutical composition, and a health functional food containing the ultrasonic extract of safflower seed as an active ingredient.

Hereinafter, the present invention will be described in detail.

The composition of the present invention contains safflower seed ultrasonic extract as an active ingredient.

The safflower seed acts to lower the concentration of cholesterol in the blood, so it is effective in the prevention and treatment of circulatory diseases such as arteriosclerosis, hyperlipidemia, and high blood pressure, and it has a therapeutic effect on fractures, osteoporosis, osteodystrophy, etc. by strengthening bones.

The safflower seeds are extracted through sonication under an extraction solvent, and specifically, the safflower seeds and the extraction solvent are mixed in a weight ratio of 1: 5 to 25, preferably 1: 10 to 20, and 20 to 50 kHz, preferably 30 to It is treated with an ultrasonicator with a frequency of 40 kHz and a power of 50 to 700 W, preferably 150 to 400 W, at 20 to 85° C., preferably 60 to 80° C. for 5 to 30 minutes, preferably 10 to 20 minutes.

When extracting safflower seeds, in the case of hot water extraction or alcohol extraction such as ethanol, not ultrasonic extraction, not only a small amount of active ingredients are extracted, but also antioxidant, UV blocking and blue light blocking effects may be low.

When the weight ratio of the safflower seed to the extraction solvent is out of the above range, the active ingredient of the safflower seed may be extracted in a small amount in the extract.

In addition, when the frequency and power of the ultrasonicator are less than the lower limit, the active ingredient of safflower seed may be extracted in a small amount, and if it exceeds the upper limit, other substances in addition to the active ingredient are extracted in a large amount, thereby reducing the effect.

In addition, when the extraction temperature and extraction time are less than the lower limit, the active ingredient of safflower seeds can be extracted in a small amount, and when it exceeds the upper limit, toxic substances may be generated.

The extraction solvent for extracting the extract is water, a lower alcohol having 1 to 4 carbon atoms, ethylene glycol, ethyl ether, or a mixed solvent thereof. As the lower alcohol, 20 to 99% by volume of an aqueous solution of methanol, ethanol, butanol or propanol may be used, and preferably, an aqueous solution of 50 to 80% by volume of ethanol may be used for excellent antioxidant, UV protection and blue light blocking effects. there is.

The ultrasonic extract of safflower seeds of the present invention may be used in health functional foods in addition to cosmetic compositions.

In addition, the present invention provides a method for preparing an ultrasonic extract of safflower seeds using a response surface analysis method.

The method for producing an ultrasonic extract of safflower seeds using the reaction surface analysis method of the present invention is (A) water, a lower alcohol having 1 to 4 carbon atoms, or an extraction solvent that is a mixed solvent thereof, an extraction temperature of 20 to 85 ℃, 10 to 60 minutes Obtaining experimental values for total polyphenols in the supernatant obtained by centrifugation after extraction with an ultrasonicator of a frequency of 20 to 50 kHz and a power of 50 to 700 W under an extraction time of ; (B) deriving a quadratic regression model represented by the following [Equation 1] using the experimental value of step (A); (C) verifying reliability by performing analysis of variance (ANOVA) on the regression model derived in step (B), and obtaining a response surface curve for the extraction conditions; (D) obtaining an experimental value for the total flavonoid content of the supernatant obtained in step (A); (E) deriving a quadratic regression model represented by the following [Equation 2] using the experimental value of step (D); (F) verifying reliability by performing analysis of variance (ANOVA) on the quadratic regression model derived in step (E), and obtaining a response surface curve for the extraction conditions; (G) obtaining an experimental value for the DPPH radical scavenging ability of the supernatant obtained in step (A); (H) deriving a quadratic regression model represented by the following [Equation 3] using the experimental values of the step (G); (I) verifying reliability by performing analysis of variance (ANOVA) on the quadratic regression model derived in step (H), and obtaining a response surface curve for the extraction conditions; and (J) predicting common optimization conditions for total polyphenol content, total flavonoid content, and DPPH radical scavenging ability by superimposing the response surface curves obtained in steps (C), (F) and (I) including; Ultrasonic extraction of safflower seeds using these predicted conditions shows excellent effects on DPPH radical scavenging ability, total polyphenol content and total flavonoid content.

[Equation 1]

Y _TPC =-4.81776-0.015551X ₁ +0.28967X ₂ +0.21412X ₃ +6.72730E-005X ₁ X ₂ +6.01865E-004X ₁ X ₃ -1.00677E-003X ₂ X ₃ -1.23645E-004X ₁ ² - 2.01372 E-0.03X ₂ ² -1.42821E-003X ₃ ²

[Equation 2]

Y _TFC =-0.37372+9.55406E-003X ₁ +9.20547E-003X ₂ +2.05361E-003X ₃ -9.72712E-005X ₁ X ₂ -5.20451E-005X ₁ X ₃ +5.37565E-005X ₂ X ₃ -1.77861E -005X ₁ ² -4.02178E-005X ₂ ² -1.79762E-005X ₃ ²

[Equation 3]

Y _DPPH =-16.21374-0.19557X ₁ +0.64596X ₂ +1.96052X ₃ +5.50661E-004X ₁ X ₂ +2.93686E-003X ₁ X ₃ -3.62518E-003X ₂ X ₃ +4.25974E-004X ₁ ² -2.87540 E-003X ₂ ² -0.013343X ₃ ²

In Equations 1 to 3, Y _TPC is the predicted value of the total polyphenol content (mg GAE / g DM), Y _TFC is the predicted value of the total flavonoid content (mg QE / g DM), Y _DPPH is the predicted value of the DPPH radical scavenging ability ( %), X ₁ is the extraction time, X ₂ is the extraction temperature, and X ₃ is the concentration of the extraction solvent.

The term 'extract' used in the present invention includes an extract obtained by extracting the components contained in safflower seeds using the solvent, a fraction fractionated therefrom, a concentrate obtained by additionally concentrating these extracts or fractions, and a purified or separated product. , is used in the sense of including a dried product of the extract, fraction, concentrate or purified product or a powder obtained by pulverizing the same.

For the production of the purified product, various additionally performed, such as passing through an ultrafiltration membrane having a molecular weight cut-off value, or separation by various chromatography (those prepared for separation according to size, charge, hydrophobicity or affinity) A purification method may be added.

Meanwhile, in the present specification, the term 'contained as an active ingredient' means including an amount sufficient to achieve the efficacy or activity of the ultrasonic extract of safflower seeds. For example, the ultrasonic extract of safflower seeds is used at a concentration of 10 to 1500 μg/ml, preferably 100 to 1000 μg/ml. Since the ultrasonic extract of safflower seed is a natural product and there is no side effect to the human body even when used in excess, the upper limit of the quantitative upper limit of the ultrasonic extract of safflower seed contained in the composition of the present invention can be selected and carried out by those skilled in the art within an appropriate range.

In the cosmetic composition of the present invention, in addition to the above cosmetic composition, other ingredients commonly formulated in cosmetics may be blended as needed. Examples of these blending ingredients include oil and fat ingredients, moisturizing agents, emollients, surfactants, organic and inorganic pigments, Organic powder, UV absorber, preservative, disinfectant, antioxidant, pH adjuster, alcohol, colorant, flavoring agent, blood circulation promoter, cooling agent, limiting agent, purified water, water-soluble vitamin, fat-soluble vitamin, polymer peptide, polymer polysaccharide, sphingolipid and seaweed an extract, etc. are mentioned.

The cosmetic composition of the present invention may be prepared in the form of emulsified formulations and solubilized formulations commonly used in the art.

In addition, the ingredients included in the cosmetic composition of the present invention may include ingredients commonly used in cosmetic compositions in addition to the ingredients as active ingredients, for example, conventional adjuvants such as stabilizers, pigments and natural fragrances; It may further include a carrier.

Products to which the composition of the present invention can be added include, for example, mist, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, sunscreen cream , moisture cream, hand cream, foundation, essence, nourishing essence, mask pack, press powder, loose powder, eye shadow, etc., and soap, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser.

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

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 particular, in the case of a spray, additional chlorofluorohydrocarbon, propane , butane or propellants such as dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylglycol oil, glycerol fatty ester, polyethylene glycol or fatty acid ester of sorbitan.

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

In addition, the present invention provides a health functional food composition containing safflower seed ultrasonic extract as an active ingredient.

Health functional food is a food prepared by adding ultrasonic safflower seed extract to food materials such as beverages, teas, spices, gum, and confectionery, or by encapsulating, powdering, or suspension, etc. However, unlike general drugs, it has the advantage of not having side effects that may occur when taking the drug for a long time using food as a raw material. The health functional food of the present invention obtained in this way is very useful because it can be ingested on a daily basis. The amount of added safflower seed ultrasonic extract in such health functional food cannot be uniformly defined as it varies depending on the type of health functional food, but it can be added within a range that does not impair the original taste of the food, and is usually 0.01 to 50% by weight, preferably 0.1 to 20% by weight. In addition, in the case of a health functional food in the form of pills, granules, tablets or capsules, it is usually added in an amount of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the health functional food of the present invention may be in the form of a pill, tablet, capsule or beverage.

In addition, the present invention provides a novel use of the ultrasonic extract of safflower seeds for the prevention or treatment of skin cancer, or for the manufacture of a medicament for animals.

The 'pharmaceutical composition' or 'medicine' may further include suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions, in addition to the ultrasonic extract of safflower seed as an active ingredient.

The 'carrier' is a compound that facilitates the addition of the compound into a cell or tissue. The 'diluent' is a compound that is diluted in water to not only stabilize the biologically active form of the compound of interest, but also to dissolve the compound.

The carrier, excipient and diluent are not particularly limited, but for example, lactose, glucose, sugar, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose , methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate and mineral oil.

The amount of the pharmaceutical composition, medicine, pharmaceutical composition for animals or veterinary medicine may vary depending on the age, sex, and weight of the patient or animal to be treated, and above all, the condition of the subject to be treated, a specific category of the disease to be treated, or It will depend on the type, route of administration, and the nature of the therapeutic agent used.

The pharmaceutical composition, medicament, pharmaceutical composition for animals or medicament for animals is appropriately selected according to the absorption rate of the active ingredient in the body, the rate of excretion, the age and weight of the patient or the animal to be treated, sex and condition, the severity of the disease to be treated, etc. , It is generally preferred to administer 0.1 to 1,000 mg/kg, preferably 1 to 500 mg/kg, more preferably 5 to 250 mg/kg, and most preferably 10 to 100 mg/kg per day. The unit dosage form formulated in this way can be administered several times at regular time intervals as needed.

The pharmaceutical composition, medicament, pharmaceutical composition for animals or medicament for animals may be administered individually as a prophylactic or therapeutic agent or may be administered in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents.

The pharmaceutical composition, medicine, pharmaceutical composition for animals or medicine for animals may be formulated into oral dosage forms such as powders, granules, tablets, capsules, troches, suspensions, emulsions, syrups, aerosols, etc. there is. In the case of formulation, it can be prepared using a diluent or excipient such as a filler, extender, binder, wetting agent, disintegrant, surfactant, etc. commonly used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, troches, and the like, and such solid preparations include at least one excipient to the compound, for example, starch, calcium carbonate, sugar or lactose, gelatin. It can be prepared by mixing and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid formulations for oral use include suspensions, solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. .

The method for treating skin cancer is administering the composition to a human or non-human animal, particularly a mammal, for example, administering the composition to a subject to be treated, which is skin cancer.

The dosage, administration method, and frequency of administration for the treatment may refer to the dosage, administration method, and frequency of administration of the pharmaceutical composition, medicine, pharmaceutical composition for animals or medicament for animals.

Hereinafter, preferred examples are presented to help the understanding of the present invention, but the following examples are merely illustrative of the present invention, and it will be apparent to those skilled in the art that various changes and modifications are possible within the scope and spirit of the present invention, It goes without saying that such variations and modifications fall within the scope of the appended claims.

Examples 1 to 17.

After drying the safflower seeds at 60 ℃ in a hot air dryer (FC 49, Lab house, Korea) for 16 hours, it was confirmed that the weight did not decrease any more, and after pulverizing the dried sample with a grinder (HMF-3000S, Hanil, Korea), 40-100 It was passed through a mesh filter and stored in a desiccator. The pulverized safflower seeds and an aqueous ethanol solution having different concentrations were mixed in a weight ratio of 1:20, put into an ultrasonicator (JAC Ultrasinic, Hwaseng, Korea), and then extracted using ultrasonic waves (40 kHz, 200 W). The extracted extract was subjected to solid-liquid separation (4 °C, 5000 rpm, 3 minutes) using a centrifuge, and only the supernatant was used.

At this time, each extract was obtained by varying the extraction time, extraction temperature, and ethanol concentration of the extraction process.

Comparative Example 1. Ethanol extract

Safflower seeds and 80% by volume of an aqueous ethanol solution were mixed in a weight ratio of 1:20 and extracted at 80° C. for 45 minutes to obtain an ethanol extract of safflower seeds.

Comparative Example 2. Methanol extract

A methanol extract of safflower seed was obtained in the same manner as in Comparative Example 1, except that 50% by volume of an aqueous methanol solution was used instead of a 50% by volume aqueous solution of ethanol.

<Test Example>

DPPH scavenging ability, total polyphenols, and total flavonoids, which are antioxidant indicators, were measured using the extracts prepared according to Examples 1 to 17 and Comparative Examples 1 to 2.

Test Example 1. Measurement of DPPH radical scavenging activity (DSA), total polyphenol content (TPC), and total flavonoid content (TFC)

5 steps of -1.68, -1, 0, 1 and 1.68 for extraction time (X ₁ ), extraction temperature (X ₂ ), and solvent concentration (X ₃ ) using central composite design (CCD) The experimental range was designed by coding with , and experimental values were obtained by testing the following 19 conditions.

1-1. DPPH radical scavenging activity (%): The antioxidant activity was measured by measuring 2,2-diphenyl-1- picrylhydrazyl (DPPH) radical scavenging activity. As a relatively stable free radical, DPPH can be reduced by ascorbic acid, tocopherol, etc. and its antioxidant activity can be measured simply by using the principle that dark purple is discolored. After diluting the sample and mixing 0.25 mL of the diluted sample with 1.25 mL of 0.1 M DPPH, it was stored in the dark at room temperature for 20 minutes, and then absorbance was measured at 517 nm. Ascorbic acid (CAS 50-81-7, Sigma, USA) was used as a control, and the same conditions as the samples were used. The radical scavenging ability was calculated according to the following [Equation 4].

[Equation 4]

Radical scavenging activity (%)=[1-(absorbance of sample / absorbance of control)]X100

1-2. Total polyphenol content (mg GAE/g DM) : It was determined by modifying the Folin-Denis method. To 0.14 mL of the extract obtained under each condition, 0.7 mL of 0.2 N Folin-Ciocalteu solution was added, and the reaction solution was left at room temperature for 8 minutes, then 0.56 mL of 7.5% Na₂CO₃ was added and reacted at room temperature for 1 hour. Then, the absorbance was measured at 765 nm using a spectrophotometer (Optizen 2120UV, KLab. Ltd., DaeJun, Korea), and a calibration curve was prepared using galic acid (Sigma-Aldrich, Minneapolis, USA) as a standard material, and the The TPC content was expressed as mg gallic acid equivalent (GAE)/g dry matter (DM).

1-3. Total flavonoid content (mg QE/g DM): A modified method of Zhishen et al. was used. TFC concentration was measured by measuring absorbance based on the principle that yellow color develops when alkali is applied to flavonoids. To 0.5 mL of each sample, 2.5 mL of distilled water and 1.5 mL of 99.5% (v/v) ethanol were added, then 0.1 mL of 1 M potassium acetate and 0.1 mL of 10% aluminum chloride were added, stirred, and left at room temperature for 30 minutes. Absorbance was measured at 415 nm, and a calibration curve was obtained by diluting quercetin (Sigma-Aldrich, Minneapolis, USA) to 25, 50, 100, 200 ug/mL to determine the flavonoid content in mg quercetin equivalent (QE)/g dry matter (DM) indicated as

division extraction time
(min) extraction temperature
(℃) ethanol concentration
(volume%) TPC (mg GAE/g DM) TFC (mg QE/g DM) DPPH scavenging ability (%) Example 1 15.0 40.0 20 5.94771 0.121412 30.8370 Example 2 45.0 40.0 20 5.61350 0.133702 25.5507 Example 3 15.0 80.0 20 6.95034 0.255068 39.2070 Example 4 45.0 80.0 20 6.65327 0.278113 37.9956 Example 5 15.0 40.0 80 8.93703 0.149833 70.9251 Example 6 45.0 40.0 80 9.64258 0.195922 74.3392 Example 7 15.0 80.0 80 7.47983 0.539985 74.0088 Example 8 45.0 80.0 80 8.30970 0.341868 74.6696 Example 9 4.77 60.0 50 9.82826 0.195922 63.9868 Example 10 55.23 60.0 50 10.65000 0.271199 66.6300 Example 11 30.0 26.36 50 7.84175 0.104513 57.9295 Example 12 30.0 93.64 50 8.23754 0.294244 65.6388 Example 13 30.0 60.0 0 5.98485 0.097026 19.1630 Example 14 30.0 60.0 99.5 7.52592 0.301925 44.1630 Example 15 30.0 60.0 50 9.71000 0.228952 65.5286 Example 16 30.0 60.0 50 10.25000 0.271199 66.2996 Example 17 30.0 60.0 50 9.49405 0.320360 66.1894 Comparative Example 1 45.0 80.0 80 3.89615 0.115610 19.4568 Comparative Example 2 45.0 80.0 80 2.71529 0.084562 13.6750

Examples 15 to 17 are experiments performed under the same conditions to confirm errors in DPPH radical scavenging activity (DSA), total polyphenol content (TPC), and total flavonoid content (TFC).

As shown in Table 1 above, the experimental value for the total polyphenol content confirmed that the extract prepared according to Example 10 was superior to other groups, and the experimental value for the total flavonoid content was excellent in Example 7 compared to other groups It was confirmed that the experimental value for the DPPH radical scavenging ability was confirmed that the extract prepared according to Example 8 was superior to that of the other groups.

In particular, in Example 13 using purified water (ultrasound), Comparative Example 1 which is an ethanol extract, and Comparative Example 2 which is a methanol extract, it was confirmed that the total polyphenol content, the total flavonoid content and the DPPH radical scavenging ability were all low.

The DPPH radical scavenging ability, the total polyphenol content and the total flavonoid content were all evenly excellent in Example 8.

Based on the experimental values, a regression equation such as [Equation 1] to [Equation 3] is derived to predict the optimal condition using Design Expert software (V.8.0, State-Ease, Inc, Minneapolis, USA) did

Test Example 2. Search for optimal conditions

division Second order polynomials ^* R ² p Total polyphenol content (mg GAE/g DM) [Equation 1]
Y _TPC =-4.81776-0.015551X ₁ +0.28967X ₂ +0.21412X ₃ +6.72730E-005X ₁ X ₂ +6.01865E-004X ₁ X ₃ -1.00677E-003X ₂ X ₃ -1.23645E-004X ₁ ² - 2.01372 E-0.03X ₂ ² -1.42821E-003X ₃ ² 0.9015 0.0086 Total flavonoid content (mg QE/g DM) [Equation 2]
Y _TFC =-0.37372+9.55406E-003X ₁ +9.20547E-003X ₂ +2.05361E-003X ₃ -9.72712E-005X ₁ X ₂ -5.20451E-005X ₁ X ₃ +5.37565E-005X ₂ X ₃ -1.77861E -005X ₁ ² -4.02178E-005X ₂ ² -1.79762E-005X ₃ ² 0.8432 0.0362 DPPH scavenging ability (%) [Equation 3]
Y _DPPH =-16.21374-0.19557X ₁ +0.64596X ₂ +1.96052X ₃ +5.50661E-004X ₁ X ₂ +2.93686E-003X ₁ X ₃ -3.62518E-003X ₂ X ₃ +4.25974E-004X ₁ ² -2.87540 E-003X ₂ ² -0.013343X ₃ ² 0.9020 0.0089

X ₁ : extraction time (min); X ₂ : extraction temperature (℃); X ₃ : ethanol concentration (%); R ² : Coefficient of Determination; p : probability value

TPC (mg/g) TFC (mg/g) DPPH Sum of Squares F
value P
value Sum of Squares F
value P
value Sum of Squares F
value P
value Model 37.11 7.12 0.0085 0.16 4.18 0.0362 4929.65 7.16 0.0083 X ₁ 0.38 0.66 0.4431 7.188E-006 1.659E-006 0.9687 101.3 1.32 0.2875 x ₂ 4,894E-004 8.449E-004 0.9776 0.094 21.70 0.0023 0.3 3.916E-003 0.9519 X ₃ 10.22 17.65 0.0040 0.045 10.38 0.0146 3010.7 39.37 0.0004 X ₁ X ₂ 3.258E-003 5.626E-003 0.9423 6.812E-003 1.57 0.2502 0.22 2.855E-003 0.9589 X ₁ X ₃ 0.59 1.01 0.3477 4.388E-003 1.01 0.3478 37.85 0.49 0.5045 X ₂ X ₃ 2.92 5.04 0.0596 8.323E-003 1.92 0.2083 13.97 0.18 0.6819 X ₁ ² 8.746E-003 0.015 0.9057 1.810E-004 0.042 0.8439 14.95 0.2 0.6717 X ₂ ² 7.33 12.66 0.0092 2.924E-003 0.67 0.4384 0.1 1.357E-003 0.9716 X ₃ ² 18.11 31.27 0.0008 2.869E-003 0.66 0.4426 1581 20.67 0.0026

2-1. Exploring conditions affecting total polyphenol content (TPC)

The experimental values for the total polyphenol content (TPC) measured according to the conditions derived using the central composite design (CCD), one of the response surface analysis methods, are shown in [Table 1].

The quadratic regression equation according to the measured total polyphenol content (TPC) can be found in [Table 2], and the R ² value, the coefficient of determination of the above regression equation, is closer to 1, the more suitable the model. admit that When the significance of the result was evaluated through Analysis of variance (ANOVA) based on the value obtained as the measured value, the R ² value for the total polyphenol content was 0.9015, and the function derived from it was suitable for predicting the total polyphenol content considered to be a model. In addition, the corresponding p -value was 0.0086, and significance was statistically verified at the level of p > 0.05, and the p-value of each independent variable was the ethanol concentration ( p = 0.0040), extraction time ( p = 0.4431), and extraction temperature ( p = 0.9776) affects the result value in that order, so it can be judged that the ethanol concentration is the main factor affecting the total polyphenol content result compared to other independent variables.

This can also be confirmed through the univariate curve [Fig. 1], which is visualized as an optimization graph by changing one variable while fixing two values of the independent variables to the median value based on the derived regression equation. . [Fig. 2] is a three-dimensional response surface curve visualized by fixing one independent variable to the median value and then changing the remaining independent variables at the same time.

It can be confirmed that the extraction temperature and the ethanol concentration sharply increase and then decrease through what is shown in [FIG. 1], and it can be confirmed that the total polyphenol content also increases as the extraction temperature increases through that shown in [FIG. 2A]. It was confirmed through [Fig. 2B] that the total polyphenol content also increased and then rapidly decreased as the extraction time and ethanol concentration increased. In addition, compared to the ethanol concentration, the total polyphenol content showed a maximum value at the median value in the extraction time and showed a tendency to decrease rapidly.

2-2. Exploring conditions affecting total flavonoid content (TFC)

The experimental values for the total flavonoid content (TFC) measured according to the conditions derived using the central composite design (CCD), one of the response surface analysis methods, are shown in [Table 1].

The quadratic regression equation according to the measured total flavonoid content (TFC) can be found in [Table 2], and the R ² value, the coefficient of determination of the regression equation above, is a model with better fit as it is closer to 1. admit it When the significance of the results was evaluated through Analysis of Variance (ANOVA) based on the values obtained as measured values, the R ² value for the total flavonoid content was determined to be 0.8432, which is a model suitable for TFC prediction. . In addition, the corresponding p -value was 0.0362, and the significance was statistically verified at the level of p > 0.05. The p -value of each independent variable showed the highest significance in the extraction time ( p = 0.0023), and the ethanol concentration ( The significance of p = 0.0146) and extraction temperature ( p = 0.9687) was confirmed to be low compared to the extraction time. Therefore, it can be determined that extraction time is the main factor affecting the result of total flavonoid content compared to other independent variables.

[Figure 3] is a univariate curve that can confirm the change in the total flavonoid content according to the change of one independent variable by fixing two independent variables among the three independent variables of extraction time, extraction temperature, and ethanol concentration to the median 0, [Fig. 4] is a three-dimensional response surface curve visualized by fixing one independent variable to the median value and then changing the remaining independent variables at the same time.

As shown in [Fig. 3], when the two independent variables were fixed at the median 0 and the change of one variable was confirmed, the main factor that had the greatest effect on the total flavonoid content was the extraction time.

As shown in [Fig. 4], [Fig. 4A] and [Fig. 4B] showing the effect of extraction time, it was confirmed that the total flavonoid content increased as the extraction time increased. In addition, in [Fig. 4C], which did not show the effect of extraction time, the total flavonoid content was maximally reduced at a constant extraction temperature and ethanol concentration, and then decreased.

2-3. Exploration of conditions affecting DPPH radical scavenging capacity (DSA)

Experimental values for DPPH radical scavenging ability (DSA) measured according to conditions derived using central composite design (CCD), one of the response surface analysis methods, are shown in [Table 1].

The quadratic regression equation according to the measured DPPH radical scavenging ability (DSA) can be found in [Table 2], and it is recognized that the closer the R ² value of the regression equation above is to 1, the more suitable the model is. When the significance of the result was evaluated through analysis of variance (ANOVA) based on the value obtained as the measured value, the R ² value for DPPH radical scavenging ability (DSA) was 0.9020, indicating that the derived function is a suitable model for TFC prediction. is judged In addition, the corresponding p -value was 0.0362, which was statistically verified for significance at the level of p > 0.05.

Also, when the p -value of each independent variable was checked, the extraction time ( p = 0.2875), extraction temperature ( p = 0.9519), and ethanol concentration ( p = 0.0004) had the slightest effect on the results, but the extraction time Also, there is no significant difference. Therefore, it can be seen that the effect of the ethanol concentration is the largest, and it can be confirmed that the ethanol concentration curve is sharp through the univariate curve [Fig. 5], which is the same as the interpretation according to the p value. In addition, after fixing one independent variable to the median value, the three-dimensional response surface curve visualized by changing the remaining independent variables at the same time is shown in FIG. 6 .

As shown in [Fig. 6], [Fig. 6A], which shows the effect of extraction time and extraction temperature, shows that the curve is gentle, so that the effect of the two independent variables does not have a significant difference in the DPPH radical scavenging ability (DSA) result value. could check In addition, it can be confirmed that the DPPH radical scavenging activity (DSA) increases as the ethanol concentration increases through [FIG. 6B] and [FIG. 6C], and it can be confirmed that the ethanol concentration gives a significant difference to the result value.

2-4. Optimal condition search

In the ultrasonic extract of safflower seed, the common optimization conditions of three variables were predicted by superimposing the response surface curves using Design Expert based on the results of individual optimization of antioxidants.

At this time, in order to secure the optimum point of inhibitory activity of DSA, TPC, and TFC while fixing the ethanol concentration among the independent variables, and to secure economic feasibility by minimizing the cost during the extraction process, the point with the lowest values of extraction time and extraction temperature was set as the optimum point. was selected. At this time, the optimal extraction conditions for ultrasonic extraction of safflower seeds were predicted to be extracted for 35.38 minutes at a temperature of 79.38 ° C in an ethanol concentration of 80.0%, and the DPPH radical scavenging ability as a dependent variable was 68.83%, and the total polyphenol content was 8.27 mg GAE/g DM, Total flavonoid content was found to be 0.39 mg QE/g DM.

Test Example 3. UV protection rate

UV A (320-400 nm) and UV B (290-320 nm) wavelengths using a UV/Vis spectrophotometer to measure the UV blocking rate of the ultrasonic extracts of safflower seeds prepared according to Examples 8, 13 and Comparative Examples. The blocking rate was measured in the range, and the absorbance was measured by comparing it with the UV blocking ability of distilled water and tannin 10 mg/mL, a representative polyphenol.

division Example 8 Example 13 Comparative Example 1 Comparative Example 2 UV-A blocking rate (%)
Distilled water vs. 27.0 25.7 15.0 13.1 UV-B blocking rate (%)
Distilled water vs. 42.1 32.7 23.8 31.2 UV-A blocking rate (%)
tannin contrast 15.45 19.43 8.1 11.5 UV-B blocking rate (%)
tannin contrast 9.93 2.74 2.4 0.4

As shown in Table 4 above, the ultrasonic wave extract of safflower seed prepared according to Example 8 of the present invention has a UV absorption rate compared to Example 13 using purified water (ultrasound), Comparative Example 1, which is an ethanol extract, and Comparative Example 2, which is a methanol extract. Because it is high, it is effective in blocking UV rays that come in contact with the skin.

That is, since the cosmetic composition applied to the skin absorbs ultraviolet rays, there is almost no ultraviolet rays that directly touch the skin.

Test Example 4. Blue light blocking rate measurement

In order to evaluate the blocking rate against blue light for the ultrasonic extracts of safflower seeds prepared according to Examples 8, 13 and Comparative Examples, the UV spectrum was measured and the blocking rate was measured using a spectrophotometer (Optizen 2120UV, Mecasys Co., Korea). did. The absorbance of the ultrasonic extract of safflower seeds prepared according to Examples 8, 13 and Comparative Example was measured in a wavelength range of 380 to 500 nm using a UV spectrophotometer, and the absorbance was measured with distilled water and a representative polyphenol, tannin 10 mg/mL. Compared with the blocking ability, the increase/decrease value of the blue light blocking rate compared to tannin was measured.

division Example 8 Example 13 Comparative Example 1 Comparative Example 2 Blue light blocking rate (%) compared to distilled water 31.6 25.7 19.65 10.77 Blue light blocking rate (%)
tannin contrast 35.05 29.71 22.32 13.36

As shown in Table 5 above, the safflower seed ultrasonic extract prepared according to Example 8 of the present invention has a blue light blocking rate compared to Example 13 using purified water (ultrasound), Comparative Example 1 which is an ethanol extract, and Comparative Example 2 which is a methanol extract. Because it is high, it is effective in blocking blue light that touches the skin.

That is, since the cosmetic composition applied to the skin blocks blue light, there is almost no blue light that directly touches the skin.

Test Example 5. Toxicity to skin cancer cells

Human melanoma cell viability was performed by the MTT staining method (J Immunol Methods, 141, 15-22, 1991). Human melanoma cells were seeded in a 96-well plate at a density of 2X10 ⁴ cells/well, and the experiment was performed after stabilization for 24 hours. The medium volume of each well was made equal to 0.1 mL. To the experimental group, each extract was dissolved in dimethyl sulfoxide (DMSO) and added at a concentration of 1, 2, 4, and 8 mg/mL. In the control group, only DMSO was added without adding the extract and cultured for 24 hours. At this time, the final DMSO content was set to 0.5%. After 24 hours, 10 μl of MTT (5 mg MTT/1 mL in PBS) was added to the medium, and human melanoma cells were further cultured for 2 hours. After removing the medium from each well, 100 μl DMSO was added and mixed by pipetting to dissolve the reduced MTT crystals. Relative human melanoma cell viability was calculated by measuring the emitted fluorescence by scanning each well with a microplate reader (Powerwave XS, Bio-tek) with a 540 nm filter.

Category (Unit: %) mg/ml 0 One 2 4 8 Example 8 100 64.645 48.283 31.769 17.639 Example 13 100 70.001 68.690 51.430 53.051 Comparative Example 1 100 75.725 73.882 72.057 69.896 Comparative Example 2 100 81.569 79.512 78.611 77.498

As shown in Table 6 above, the safflower seed ultrasonic extract prepared according to Example 8 of the present invention was melanoma cells compared to Example 13 using purified water (ultrasound), Comparative Example 1 which was an ethanol extract, and Comparative Example 2 which was a methanol extract. It was confirmed that the mortality rate of Accordingly, the ultrasonic extract of safflower seed prepared according to Example 8 of the present invention can prevent, treat or improve skin cancer.

Hereinafter, a formulation example of a composition containing the extract of the present invention will be described, but the present invention is not intended to limit it, but to describe it in detail.

Formulation Example 1: Preparation of powder

20 mg of extract powder of Example 8

Lactose 100 mg

talc 10 mg

The above ingredients are mixed and filled in an airtight bag to prepare a powder.

Formulation Example 2: Preparation of tablets

10 mg of extract powder of Example 8

100 mg cornstarch

Lactose 100 mg

2 mg magnesium stearate

After mixing the above ingredients, tablets are prepared by tableting according to a conventional manufacturing method of tablets.

Formulation Example 3: Preparation of capsules

10 mg of extract powder of Example 8

3 mg of crystalline cellulose

Lactose 14.8 mg

0.2 mg magnesium stearate

According to a conventional capsule preparation method, the above ingredients are mixed and filled in a gelatin capsule to prepare a capsule.

Formulation Example 4: Preparation of granules

1,000 mg of extract powder of Example 8

appropriate amount of vitamin mixture

70 μg vitamin A acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

0.5 mg of vitamin B6

0.2 μg of vitamin B12

Vitamin C 10 mg

Biotin 10 μg

Nicotinamide 1.7 mg

50 μg of folic acid

Calcium pantothenate 0.5 mg

Mineral mixture appropriate amount

ferrous sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

potassium phosphate monobasic 15 mg

Dibasic calcium phosphate 55 mg

Potassium citrate 90 mg

100 mg of calcium carbonate

Magnesium chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture is mixed with ingredients suitable for health functional food in a preferred embodiment, but the mixing ratio may be arbitrarily modified, and the ingredients are mixed according to a conventional health functional food manufacturing method. Next, the granules can be prepared and used in the preparation of a health functional food composition according to a conventional method.

Formulation Example 5: Preparation of beverage formulations

1,000 mg of extract powder of Example 8

1,000 mg citric acid

100 g of oligosaccharides

2 g of plum concentrate

1 g taurine

Add purified water to total 900 mL

After mixing the above ingredients according to a conventional beverage manufacturing method, after stirring and heating at 85 ° C. for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized, and then refrigerated. used in the manufacture of functional beverage compositions.

A preparation example of a cosmetic composition suitable for applying the present invention will be presented.

Preparation 6: lotion

Preparation examples of the lotion among the cosmetics containing the ultrasonic extract of Example 8 are shown in Table 7 below.

ingredient content (wt%) Extract of Example 8 5.0 glycerin 6.0 1,3-butylene glycol 3.0 PG 1500 1.0 allantoin 0.1 DL-Panthenol 0.3 EDTA-2Na 0.02 Benzophenone-9 0.04 Sodium Hyaluronate 5.0 ethanol 10.0 Polysorbate 20 0.2 preservatives, fragrances, colors a very small amount Distilled water remaining amount Sum 100

Preparation Example 7: Lotion

Preparation examples of the lotion in the cosmetic containing the ultrasonic extract of Example 8 are shown in Table 8 below.

ingredient content (wt%) Extract of Example 8 3.0 propylene glycol 6.0 glycerin 4.0 triethanolamine 1.2 tocopheryl acetate 3.0 liquid paraffin 5.0 squalane 3.0 Macadamin Nut Oil 2.0 Polysorbate 60 1.5 Sorbitan sesquioleate 1.0 Carboxyvinyl Polymer 1.0 preservatives, fragrances, colors a very small amount Distilled water remaining amount Sum 100

Preparation 8: Nourishing Cream

Preparation examples of the nutritional cream in the cosmetic containing the ultrasonic extract of Example 8 are shown in Table 9 below.

ingredient content (wt%) Extract of Example 8 1.0 lipophilic monostearate glycerin 1.5 cetearyl alcohol 1.5 stearic acid 1.0 Polysorbate 60 1.5 Sorbitan Stearate 0.6 Isostearylisostearate 5.0 squalane 5.0 mineral oil 35.0 dimethicone 0.5 Hydroxyethyl Cellulose 0.12 glycerin 6.0 triethanolamine 0.7 preservatives, fragrances, colors a very small amount Distilled water remaining amount Sum 100

Preparation Example 9: Essence

Preparation examples of the essence in the cosmetics containing the ultrasonic extract of Example 8 are shown in Table 10 below.

ingredient content (wt%) Extract of Example 8 1.5 glycerin 10.0 betaine 5.0 PEG 1500 2.0 allantoin 0.1 DL-Panthenol 0.3 EDTA-2Na 0.02 Benzophenone-9 0.04 Hydroxyethyl Cellulose 0.1 Sodium Hyaluronate 8.0 Carboxyvinyl Polymer 0.2 triethanolamine 0.18 Octyldodecanol 0.3 Octyldodeces-16 0.4 ethanol 6.0 preservatives, fragrances, colors a very small amount Distilled water remaining amount Sum 100

Preparation 10: Emulsion for mask pack

Preparation examples of the emulsion for a mask pack among cosmetics containing the ultrasonic extract of Example 8 are shown in Table 11 below.

ingredient content (wt%) Extract of Example 8 1.0 polyvinyl alcohol 15.0 Cellulose Gum 0.15 glycerin 3.0 PEG 1500 2.0 cyclodextrin 0.15 DL-Panthenol 0.4 allantoin 0.1 Monoammonium glycyrrhizinate 0.3 nicotinamide 0.5 ethanol 6.0 PEG 40 hydrogenated castor oil 0.3 preservatives, fragrances, colors a very small amount Distilled water remaining amount Sum 100

Claims (10)

Contains safflower seed ultrasonic extract as an active ingredient,
The safflower seed ultrasonic extract is an antioxidant, characterized in that it is obtained by processing safflower seeds and 80% by volume of an ethanol aqueous solution at an extraction temperature of 60 to 80 ° C with a frequency of 20 to 50 kHz and an ultrasonicator of 50 to 700 W power for 45 minutes. A cosmetic composition for blocking UV rays and blocking blue light. delete delete delete delete Contains safflower seed ultrasonic extract as an active ingredient,
The safflower seed ultrasonic extract is an antioxidant, characterized in that it is obtained by processing safflower seeds and 80% by volume of an ethanol aqueous solution at an extraction temperature of 60 to 80 ° C with a frequency of 20 to 50 kHz and an ultrasonicator of 50 to 700 W power for 45 minutes. Health functional food for UV protection and blue light blocking. (A) Safflower seeds in water, an extraction solvent that is a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, an extraction temperature of 20 to 85 ° C., a frequency of 20 to 50 kHz and a frequency of 50 to 700 W under an extraction time of 10 to 60 minutes obtaining experimental values for total polyphenols in the supernatant obtained by centrifugation after extraction with a power sonicator;
(B) deriving a quadratic regression model represented by the following [Equation 1] using the experimental value of step (A);
(C) verifying reliability by performing analysis of variance (ANOVA) on the regression model derived in step (B), and obtaining a response surface curve for the extraction conditions;
(D) obtaining an experimental value for the total flavonoid content of the supernatant obtained in step (A);
(E) deriving a quadratic regression model represented by the following [Equation 2] using the experimental value of step (D);
(F) verifying reliability by performing analysis of variance (ANOVA) on the quadratic regression model derived in step (E), and obtaining a response surface curve for the extraction conditions;
(G) obtaining an experimental value for the DPPH radical scavenging ability of the supernatant obtained in step (A);
(H) deriving a quadratic regression model represented by the following [Equation 3] using the experimental values of the step (G);
(I) verifying reliability by performing analysis of variance (ANOVA) on the quadratic regression model derived in step (H), and obtaining a response surface curve for the extraction conditions; and
(J) Predicting common optimization conditions for total polyphenol content, total flavonoid content, and DPPH radical scavenging ability by superimposing the response surface curves obtained in steps (C), (F) and (I) above step; including,
The extraction conditions according to the common optimization conditions for the DPPH radical scavenging ability, the total polyphenol content and the total flavonoid content are an extraction time of 35.38 minutes, an extraction temperature of 79.38 ° C. and an ethanol concentration of 80% by volume Antioxidation using a reaction surface analysis method A method for preparing an ultrasonic extract of safflower seeds;
[Equation 1]
Y _TPC =-4.81776-0.015551X ₁ +0.28967X ₂ +0.21412X ₃ +6.72730E-005X ₁ X ₂ +6.01865E-004X ₁ X ₃ -1.00677E-003X ₂ X ₃ -1.23645E-004X ₁ ² - 2.01372 E-0.03X ₂ ² -1.42821E-003X ₃ ²
[Equation 2]
Y _TFC =-0.37372+9.55406E-003X ₁ +9.20547E-003X ₂ +2.05361E-003X ₃ -9.72712E-005X ₁ X ₂ -5.20451E-005X ₁ X ₃ +5.37565E-005X ₂ X ₃ -1.77861E -005X ₁ ² -4.02178E-005X ₂ ² -1.79762E-005X ₃ ²
[Equation 3]
Y _DPPH =-16.21374-0.19557X ₁ +0.64596X ₂ +1.96052X ₃ +5.50661E-004X ₁ X ₂ +2.93686E-003X ₁ X ₃ -3.62518E-003X ₂ X ₃ +4.25974E-004X ₁ ² -2.87540 E-003X ₂ ² -0.013343X ₃ ²
In Equations 1 to 3, Y _TPC is the predicted value of the total polyphenol content (mg GAE / g DM), Y _TFC is the predicted value of the total flavonoid content (mg QE / g DM), Y _DPPH is the predicted value of the DPPH radical scavenging ability ( %), X ₁ is the extraction time, X ₂ is the extraction temperature, and X ₃ is the concentration of the extraction solvent. delete Contains safflower seed ultrasonic extract as an active ingredient,
The ultrasonic extract of safflower seeds is obtained by treating safflower seeds and 80% by volume of an aqueous ethanol solution in an extraction temperature of 60 to 80 ° C. with an ultrasonicator of a frequency of 20 to 50 kHz and a power of 50 to 700 W for 45 minutes. A pharmaceutical composition for prophylaxis or treatment. The pharmaceutical composition according to claim 9, wherein the skin cancer is melanoma.

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