KR20160143003A - The extraction procedure for iceplant's active constituent - Google Patents

Abstract

The present invention relates to a method of extracting an effective component of an ice plant for a cosmetic composition, and more particularly, to a method for extracting an effective component of an ice plant for a cosmetic composition by drying leaves and stems of a washed ice plant; Pulverizing the dried ice plant to a predetermined particle size; Immersing the powdered ice plant in an aqueous ethanol solution for a predetermined period of time, and extracting an effective component of the ice plant; Concentrating the extracted active ingredients of the ice plant under reduced pressure to remove the ethanol; And lyophilizing the active ingredient of the reduced-pressure concentrated ice plant.

Description

[Technical Field] The present invention relates to an extraction method for an ice plant for cosmetic composition,

The present invention relates to a method for extracting an effective component of an ice plant for a cosmetic composition, and more particularly, to a cosmetic composition capable of easily and easily obtaining an effective component of an ice plant having antioxidant activity, whitening activity, anti- To an active ingredient extraction method for an ice plant for a composition.

In accordance with recent natural and environmentally friendly consumption trends, effective ingredients for cosmetics have been used in cosmetics in various forms based on their usefulness as well as chemical substances as well as natural products derived from plants or animals.

In particular, the development of cosmetics using various natural materials, ranging from botanical raw materials including herbal medicines to marine raw materials, in the wind of naturalism, is coming to an era of functional cosmetics using natural products.

Overall, Korea's cosmetics technology level is the highest in the field of formulation, the most weak in the field of materials, and there is a gap of three to five years with developed countries, but the industry and academy jointly focus on developing new functional cosmetics materials. It will be enough to solve the gap and have international competitiveness.

Skin aging is divided into human aging (or chronological aging) and photoaging (or photophobia aging).

Chronological aging is a biological aging process that occurs as a result of aging, and the fine dermis is atrophied and the subcutaneous fat layer is reduced and fine wrinkles are observed.

In contrast, photophobia aging is a combination of degenerative changes and chronological aging that occur in the sun exposed areas.

In this process, rough and deep wrinkles appear, abnormally elastic fibrous material accumulates and the skin thickens like leather.

Apart from this aging process, wrinkles occur due to muscle distribution and muscle movements on the face.

The face has various muscles balanced to create a contour of the face, and the muscles are connected to the skin and closely related.

If you do not tighten the tether around your tent, you will have a wrinkle of your face when the muscle tissue that catches your skin weakens or becomes tangled as if the tent is stretched. So if you always make a face mixed with irritation on your face, you have a lump in your forehead and a thick wrinkle.

Therefore, the bad wrinkles in the wrinkles are the bad wrinkles because the wrinkles that draw the inner puncture between the eyes and the wrinkles that draw the ghosts seem to irritate the impression of the person.

Therefore, if you make the right condition of the facial muscles, the wrinkles will be better and elasticity will be generated.

In photophobia aging, the skin becomes dry, rough and thick, whereas in chronological aging, the skin becomes thinner and smoother.

In addition, there are deeper wrinkles in photophobia aging, unlike chronological aging, and there are relaxation, stained hyperpigmentation and hypopigmentation, capillary dilatation, sputum and caustic scars.

In addition, chronological aging and photophobia aging have different clinical features. The main cause of photophobia aging is ultraviolet light.

Ultraviolet rays can be divided into UV-A, B and C according to the wavelength. UV-A is less harmful to the skin than UV-B because it has little stimulus to the skin. However, if it is exposed for a long time, Of pigmentation occurs and not only deepens skin aging but also can cause skin cancer even in severe cases.

On the other hand, UV-B causes strong skin irritation, causing sunburn or skin cancer, but can not penetrate glass, so it is safe in the room.

UV-C is a ray of light with a wavelength close to that of X-rays, but it is highly carcinogenic, but it is not necessary to block it because it is blocked by the ozone layer.

As living standards improve and interest in skin beauty increases, interest in skin aging is increasing.

Although it is impossible to prevent skin aging, various attempts have been made to suppress and slow down the aging of the skin and wrinkles, which is a representative symptom thereof.

Modern medicine provides a means of alleviating the physical signs of aging, but it produces many side effects, and it is not easy to find natural substances that have no side effects and are suitable for the skin.

The skin is largely divided into three layers of epidermis, dermis and subcutaneous tissue from the outside in order from the outside. It functions to dissolve the internal organs in the body from changes in temperature and humidity, ultraviolet rays and other physical and chemical external environment.

In particular, the epidermis plays an important role in preventing water evaporation inside the human body.

The epidermis is divided into stratum corneum, granular layer, superficial layer, and basal layer in order from the outside. Cells of stratum corneum act like bricks, and intercellular lipids between keratinocytes act like mortar to constitute skin barrier.

In addition, healthy keratinocytes have a high concentration of natural moisturizing factors to help maintain moisture in the skin. For example, substances such as amino acids are water-soluble and effectively combine with water to inhibit drying of moisture from the skin.

However, as it is nowadays, there is a tendency to adjust the temperature of the air / heating due to the change of environment or life pattern, the stress caused by various stresses and environmental pollution in the social life, frequent washing according to make- Due to various causes such as aging of skin and the like, moisture of the stratum corneum is reduced, and the skin becomes dry, the surface becomes rough, the skin becomes loose, the luster is lost and the skin looks dull, and the need for skin moisturizing agent is increasing.

Accordingly, in order to solve these problems, cosmetics using natural products have been developed in recent years in order to reduce skin irritation and aging caused by various chemical substances and the like.

Natural physiologically active substances have a small adverse effect on the skin, and as the response of consumers to natural substances increases, the development value as a cosmetic raw material is further increasing.

Disclosure of the Invention The present invention has been made to solve the above-mentioned problems and to meet the needs, and has as its object to provide an ice plant having an antioxidant activity, a whitening activity, an anti-inflammatory activity, an antibacterial activity and a wrinkle- And to provide a method for extracting an effective component of an ice plant for cosmetic composition.

In the meantime, the objects of the present invention are not limited to the above-mentioned objects, and other objects not mentioned can be clearly understood by those skilled in the art from the following description.

The method for extracting an effective component of an ice plant for a cosmetic composition according to the first embodiment of the present invention includes the steps of drying leaves and stems of a washed ice plant, , A step of immersing the powdered ice plant in an aqueous ethanol solution for a predetermined period of time and then extracting an active ingredient of the ice plant, a step of removing the ethanol from the active ingredient of the extracted ice plant Concentrating under reduced pressure, and lyophilizing the active ingredient of the reduced-pressure concentrated ice plant.

The method of extracting an effective component of an ice plant for a cosmetic composition according to a second embodiment of the present invention includes the steps of drying leaves and stems of a washed ice plant, , Adding a strain to the powdered ice plant, culturing the mixture to prepare an ice plant culture liquid, centrifuging and filtering the culture liquid to prepare a filtrate from which the strain has been removed And aging the filtrate for a predetermined period of time, followed by re-filtration to obtain an extract containing the ice plant active ingredient.

Preferably, the strain is Bacillus subtilis (Bacillus subtilis), saccharose in my process celebrity bicyclic Ke (Saccharomyces S. cerevisiae , and Lactobacillus acidophilus .

Meanwhile, in order to achieve the above-mentioned object, the method of extracting an effective component of an ice plant for a cosmetic composition according to a third embodiment of the present invention comprises the steps of drying leaves and stems of a washed ice plant, , Powdering the powdered ice plant, and shaving the steamed ice plant.

According to a fourth aspect of the present invention, there is provided a method for extracting an effective component of an ice plant for a cosmetic composition, comprising the steps of: drying leaves and stem of a washed ice plant; , And extracting the active ingredient of the ice plant using the supercritical extraction method of the powdered ice plant.

The present invention has the following excellent effects.

First, an effective component of an ice plant having antioxidative activity, whitening activity, anti-inflammatory activity, antimicrobial activity and wrinkle improving activity can be easily and easily extracted. According to the embodiments of the present invention, the extracted ice plant extract has an oxidative stress And inflammation-induced skin diseases, and can be usefully used as a composition for external use on the skin and a cosmetic composition.

In addition, the extracted ice plant extract according to the embodiments of the present invention promotes differentiation of keratinocyte cells, restores skin barrier function, eliminates the pulling and roughness of the skin, and increases the elasticity, There is an excellent effect that can be usefully used.

BRIEF DESCRIPTION OF DRAWINGS FIG. 1 is an overall process diagram of a method for extracting an effective component of an ice plant for a cosmetic composition according to a first embodiment of the present invention. FIG.

The term used in the present invention is a general term that is widely used at present. However, in some cases, there is a term selected arbitrarily by the applicant. In this case, the term used in the present invention It is necessary to understand the meaning.

Hereinafter, the technical structure of the present invention will be described in detail with reference to preferred embodiments shown in the accompanying drawings.

First, FIG. 1 is an overall process diagram of an effective component extraction method of an ice plant for a cosmetic composition according to the first embodiment of the present invention.

Referring to FIG. 1, the method for extracting an effective component of an ice plant for a cosmetic composition according to the first embodiment of the present invention includes drying leaves and stems of a washed ice plant.

The ice plant is a succulent plant having a low germination rate and a low growth rate. It is originated from the Namib desert in South Africa. The main distribution areas are southern Africa, eastern and western parts, northern part, southwestern part of Australia, and western part of the Americas.

On the other hand, the transparent crystals (bladder cells) on the surface of the ice plant contain components useful for the human body such as inositol and beta carotene and various minerals. Bladder cells of the stem and leaf are shaped like ice crystals It is called ice plant.

Especially, since the ice plant contains a lot of pinitol which lowers the blood sugar level and myoinositol which suppresses the neutral fat, it is possible to use the healing food vegetable )to be.

In the first embodiment of the present invention, the leaves and stems of the ice plant are used. However, the present invention is not limited thereto. In addition to leaves and stems, various ice plants such as flowers, roots, branches, Can be used.

Meanwhile, the method for extracting an effective component of an ice plant for a cosmetic composition according to the first embodiment of the present invention includes a step of pulverizing the dried ice plant to a predetermined particle size.

In this case, the particle size of the powdered ice plant may be pulverized into various particle sizes. However, in the first embodiment, the particle size is 300 mesh or less. In order to powder the ice plant, Branch can be used, so there is no particular limitation.

Thereafter, the step of immersing the powdered ice plant in the aqueous ethanol solution for a predetermined period of time, and then extracting the active component of the ice plant is performed.

The step of extracting the active ingredient may be performed by adding the ice plant to an ethanol aqueous solution of 1 to 10 times the dry weight of the prepared ice plant powder, heating the mixture at 50 to 100 ° C for 5 to 24 hours, At a temperature of 38 DEG C for 1 to 14 days, and then the active ingredient is extracted.

In the first embodiment of the present invention, an ethanol aqueous solution is used as an extraction solvent. However, the present invention is not limited thereto, and acetone, ethylacetate, butyl acetate, An aqueous solution containing 1,3-butylene glycol may also be used.

Meanwhile, the method for extracting an effective component of an ice plant for a cosmetic composition according to the first embodiment of the present invention includes a step of concentrating under reduced pressure to remove the ethanol from an effective component of the extracted ice plant.

At this time, various steps can be used for the step of concentration under reduced pressure, but in the first embodiment of the present invention, the reaction is concentrated under reduced pressure using a rotary evaporator.

On the other hand, when 1,3-butylene glycol is used as the extraction solvent, it is difficult to remove 1,3-butylene glycol through the rotary evaporator. Therefore, the 1,3-butylene glycol is removed by using a hot-air drier at 30 to 60 ° C. so that the loss on drying is 1% (w / w).

Finally, the method for extracting an effective component of an ice plant for a cosmetic composition according to the first embodiment of the present invention includes a step of freeze-drying an active ingredient of the reduced-pressure concentrated ice plant.

At this time, the lyophilizing step may be performed through various equipment or means, so that no particular limitation is placed on the lyophilization step.

Hereinafter, a method for extracting an effective component of an ice plant for a cosmetic composition according to a second embodiment of the present invention will be described in detail.

In this regard, the method for extracting an effective component of an ice plant for a cosmetic composition according to a second embodiment of the present invention includes the steps of drying leaves and stems of a washed ice plant, and pulverizing the dried ice plant to a predetermined particle size Since these steps are the same as those described in the first embodiment, a detailed description thereof will be omitted below.

On the other hand, a method for extracting an effective component of an ice plant for a cosmetic composition according to a second embodiment of the present invention includes a step of adding a strain to the powdered ice plant, followed by culturing to prepare an ice plant culture liquid.

Herein, the strain may be a fermentative, yeast, or lactic acid bacterium. Hereinafter, embodiments of the respective strains will be described in detail.

First, when a fermenting microorganism is used as the strain (Example 2-1), Bacillus subtilis, Bacillus subtilis, Bacillus licheniformis, Bacillus megaterium, Bacillus polyfermenticus, Bacillus (genus Bacillus ) including a mixture thereof can be used, and preferably Bacillus subtilis is used.

Yeast cells can be used as the strain (Example 2-2). Yeast cells used in this process belong to eukaryotic microorganisms and have cell organelles such as cell walls, cell membranes, nuclei, mitochondria, and granules.

On the other hand, yeasts which can be used in the embodiment 2-2 of the present invention include Aureobasidium pullurans and Saccharomyces cerevisiae cerevisiae . In the case of Saccharomyces cerevisiae , it is killed at above 40 DEG C, but oleic acid and ergosterol may be added to the medium to prevent this.

(Example 2-3) The lactic acid bacteria are a kind of microorganisms which are very useful for the human body which decomposes carbohydrates such as glucose or lactose to produce organic acids such as lactic acid and acetic acid. Lactobacillus, Streptococcus, Bifidobacterium, Leuconostock, Pediococcus, Lactococcus, and mixtures thereof may be used. In the 2-3 embodiment of the present invention, it is preferable to use Lactobacillus Do.

Meanwhile, in the step of adding the strain to the powdered ice plant according to the second embodiment of the present invention and preparing a culture solution for the ice plant, the powder of the ice plant and the purified water are mixed at a ratio of 1:10, Strain sterilized at 121 占 폚 is used and the powder of the ice plant used in this step is used in an amount of 1 to 100 g based on 1 liter of the total culture.

In addition, the strain is preferably added in an amount of 1 × 10 ⁵ to 1 × 10 ⁶ cells per liter of the total culture medium. Depending on the characteristics of each strain, the strain may be cultured under ordinary conditions and aerobic or normal anaerobic conditions .

In addition, in order to further activate the culture of the fermenting bacteria, additional carbon sources (glucose, maltose, xylose, starch, etc.), nitrogen source (peptone, yease extract, ammonium sulfate and the like), trace elements (potassium phosphate, sodium phosphate, Manganese, etc.) and a pH adjuster may be added.

In addition, suitable culture conditions of the strain are cultured under aerobic or normal anaerobic conditions at 25 to 37 ° C, pH 6 to 7, preferably 1 to 14 days, and most preferably 1 to 7 days.

Meanwhile, the method for extracting an effective component of an ice plant for a cosmetic composition according to the second embodiment of the present invention comprises the steps of preparing the filtrate from which the strain has been removed by centrifuging and filtering the culture solution, After aging, re-filtration is carried out to obtain an extract containing the ice plant active ingredient.

At this time, in the step of aging the filtrate for a predetermined period of time and then re-filtering to obtain an extract containing the ice plant active ingredient, the step of primary filtration of the culture broth and the aging of the filtrate at low temperature, Can be filtered with a filtration membrane having a size of 0.25 to 0.45 mu m to remove excess ice plant powder particles and fermenting bacteria.

At this time, the filtrate separated by the primary filtration membrane can be stabilized by low-temperature aging and can be filtered again with a finer filtration membrane as compared with the primary filtration membrane, and then an effective component of the ice plant can be obtained.

On the other hand, the low-temperature aging is preferably performed for 3 to 14 days at a temperature of 0 to 4 캜, and the size of the filtration membrane is preferably 0.25 탆 or less in order to reliably remove remaining bacteria.

Hereinafter, a method of extracting an effective component of an ice plant for a cosmetic composition according to a third embodiment of the present invention will be described in detail.

The method for extracting an effective component of an ice plant for a cosmetic composition according to the third embodiment of the present invention comprises the steps of drying leaves and stems of a washed ice plant, pulverizing the dried ice plant to a predetermined particle size, And a step of shaving the steamed ice plant.

In this case, the steps of drying the leaves and stems of the washed ice plant and pulverizing the dried ice plant to a predetermined particle size are the same as those of the first embodiment described above, so a description thereof will be omitted.

Meanwhile, the step of brewing the ice plant and the step of shaving the steamed ice plant may be performed under various conditions. However, in the third embodiment of the present invention, the step of boiling at 120 to 180 ° C for 10 minutes to 2 hours .

Meanwhile, in the third embodiment of the present invention, the step of pouring the powdered ice plant may be replaced with a step of boiling the powdered ice plant or a step of baking the powdered ice plant, Boiling the ice plant may be carried out at 60-100 ° C for 30 minutes to 10 hours, and baking the powdered ice plant at 80-100 ° C for 10 minutes to 2 hours.

On the other hand, the step of shaving the steamed ice plant may be natural shredding under various conditions, so that no particular limitation is given thereto.

Hereinafter, a method for extracting an effective component of an ice plant for a cosmetic composition according to a fourth embodiment of the present invention will be described in detail.

A method for extracting an effective component of an ice plant for a cosmetic composition according to a fourth embodiment of the present invention comprises the steps of drying leaves and stems of a washed ice plant, pulverizing the dried ice plant to a predetermined particle size, And extracting an effective component of the ice plant using the supercritical extraction method.

The step of drying the leaves and stems of the washed ice plant and the step of pulverizing the dried ice plant to a predetermined particle size are the same as those of the first embodiment described above, and a description thereof will be omitted below .

Meanwhile, the method of extracting an effective component of an ice plant for a cosmetic composition according to the fourth embodiment of the present invention includes extracting an effective component of an ice plant using the supercritical extraction method for the ice plant as described above do.

At this time, the supercritical fluid used in the supercritical extraction method may be a mixed fluid in which a co-solvent is further mixed with supercritical carbon dioxide or carbon dioxide. Preferably, the supercritical fluid is used in a supercritical state under a pressure of 300 atm And is extracted with olive oil as a hawk.

Hereinafter, the effect of the extract of the active ingredient of the extracted ice plant according to the embodiments of the present invention will be described in detail.

1. Measurement of antioxidant effect DPPH Law)

The antioxidant activity was measured by Sakata's method for scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals.

That is, 0.9 ml of the solution (0.15 mM) prepared by dissolving DPPH (Sigma, USA) in ethanol and 0.1 ml of the sample solution were put in a test tube, mixed at Voltex, reacted at room temperature for 10 minutes and then absorbance was measured at 517 nm Respectively.

At this time, the control group was compared with natural-tocopherol, which is a natural antioxidant, and the free radical scavenging activity (%) was calculated. The measurement results are shown in Table 1 below.

sample Treatment concentration (%) Free radical scavenging ability (%) First Embodiment Example 2-1 Example 2-2 Example Example 2-3 Third Embodiment Fourth Embodiment Ice Plant Extract 0.0005 10.5 15.2 12.2 13.4 11.5 8.9 0.001 33.8 32.1 34.5 33.0 29.8 22.6 0.005 77.4 70.5 71.8 72.6 67.9 65.8 0.010 90.5 88.8 87.9 91.6 89.7 82.6 0.020 94.2 91.5 93.7 92.4 90.4 89.5 Natural-tocopherol 0.050 90.4

As shown in Table 1, the antioxidant activity of ice plant extracts was increased in a concentration-dependent manner, and the ice plant extract had a higher antioxidative effect than that of natural-tocopherol there was.

Among them, 0.02% concentration was the highest in the first embodiment, followed by the second to the second embodiment.

2. Measurement of whitening effect

2-1. Inhibitory effect on melanin formation

Melanoma cells (B16 melanoma cells) were dispensed in a 6-well plate at a concentration of 2 × 10 ⁶ cells per well and cells were adhered. Then, the cells were treated with ice plant extracts according to the first to fourth embodiments at concentrations not causing toxicity And cultured for 72 hours.

After culturing for 72 hours, the cells were detached with trypsin-EDTA, the number of cells was measured, and the cells were recovered by centrifugation.

The cell pellet was washed once with PBS, and 1 ml of homogenization buffer solution (50 mM sodium phosphate, pH 6.8, 1% Triton X-100, 2 mM PMSF) was added and vortexed for 5 minutes to disrupt the cells.

After centrifugation (3,000 rpm, 10 min), 1N NaOH (10% DMSO) was added to the cell filtrate to dissolve the extracted melanin, and the absorbance of melanin was measured at 405 nm using a microplate reader. Melanin was quantitated to determine the melanin The percent inhibition of production was measured.

Melanogenesis inhibition rate (%) of melanin-forming cells was calculated by the following formula (1), and the results are shown in Table 2 below.

At this time, hydroquinone, which is known to have an inhibitory effect on melanin formation, was used as a control.

[Equation 1]

Melanin formation inhibition rate (%) = [(a-b) / a] x 100

a: Amount of melanin in well without addition of sample

b: amount of melanin in the well to which the sample is added

sample
Treatment concentration
(%) Melanin inhibition rate (%) First Embodiment Example 2-1 Example 2-2 Example Example 2-3 Third Embodiment Fourth Embodiment Ice Plant Extract 0.05 16.8 10.9 11.5 14.3 10.7 11.5 0.10 33.5 27.4 30.5 31.5 27.9 29.4 0.25 61.9 48.0 60.4 58.4 44.8 45.6 0.50 81.6 64.9 70.8 77.8 71.8 69.7 Hydroquinone 100ppm 72.4

As shown in Table 2, the ice plant extract prepared according to Examples 1 to 4 significantly inhibited melanin production in melanin-forming cells in a concentration-dependent manner.

Among them, the treatment concentration was the highest in the first embodiment at the treatment concentration of 0.50%, and the lowest in the second embodiment.

2-2. Tyrosinase  Measurement of inhibitory activity

The inhibitory activity of DOPA (dihydroxyphenylalanine) or dopachrome produced by tyrosinase, a key enzyme of melanin synthesis, was measured.

In the test method, tyrosine was added to the enzyme reaction solution for a predetermined period of time, and the absorbance was measured at 475 nm, which is the absorption wavelength of the reaction product, dopachrome.

Mushroom tyrosinase inhibition assay method was used to treat 0.2 ml of Mushroom tyrosinase (110 U / ml) in a mixture of 0.2 ml of substrate solution in which 10 mM Tyrosine (L-DOPA) was dissolved in 0.1 M Potasium phosphate buffer And the reaction was carried out at 25 ° C for 5 minutes. The DOPA chrome produced in the reaction solution was measured at 475 nm. The measurement results are shown in Table 3.

Tyrosinase inhibitory activity was indicated by the rate of decrease in the degree of scratches in the samples without and with no additives.

&Quot; (2) "

Tyrosinase activity inhibition rate (%) = [(a-b) / a] x 100

a: absorbance after reaction of reaction solution without inhibitor

b: Absorbance after reaction of reaction mixture containing test sample

sample Treatment concentration
(ug / ml) Inhibition rate of tyrosinase activity (%) First Embodiment Example 2-1 Example 2-2 Example Example 2-3 Third Embodiment Fourth Embodiment Ice Plant Extract 50 11.5 10.8 13.3 10.5 11.5 8.5 100 38.7 29.8 31.5 29.8 20.5 22.3 500 70.3 41.5 61.2 49.9 48.5 48.1 1,000 85.8 52.4 72.8 64.9 57.6 61.4

As shown in the above Table 3, it can be seen that the ice plant extract prepared according to the first to fourth embodiments remarkably inhibits tyrosinase activity in a concentration-dependent manner.

Among them, the treatment concentration was the highest in the first saline at the treatment concentration of 1,000 ug / ml, followed by the second to the second saline.

In general, the tyrosinase activity was lowest in Example 2-1, indicating that the fermented product of the ice plant extract fermented by Bacillus subtilis did not effectively inhibit the melanin biosynthesis compared to the other examples.

3. Wrinkle improvement effect

3-1. Ice Plant Extract Collagenase  Effect of biosynthesis promotion

Human normal epithelial cells, fibroblasts, were inoculated on a 48-well microplate to a concentration of 1 × 10 ⁷ cells per well, and cultured in DMEM medium for 24 hours.

Serum-free DMEM medium supplemented with the ice plant extract prepared in Examples 1 to 4 and a control group were further cultured in serum-free DMEM medium without ice plant extract for 60 hours.

After the incubation, the supernatant of each well was collected and the amount of procollagen type I C-peptide (PICP) was measured by using a collagen kit and converted into ng / ml, and the amount of synthesized collagen was measured.

Collagen biosynthesis increase rate (%) was calculated according to the following formula (3), and the results are shown in Table 4 below.

&Quot; (3) "

Collagenase biosynthesis promotion rate (%) = [(a / b) -1] × 100

a: The amount of collagen in the experimental group

b: Collagen amount of control group

sample Treatment concentration
(ppm) Promotion rate of collagen biosynthesis (%) First Embodiment Example 2-1 Example 2-2 Example Example 2-3 Third Embodiment Fourth Embodiment Ice Plant Extract 5 51.5 11.2 28.7 19.7 20.4 26.8 10 79.8 35.4 62.5 30.2 52.4 61.6 50 105.4 66.5 98.7 61.5 88.7 92.5 100 121.5 75.5 114.8 80.2 105.9 100.4

As shown in Table 4, the ice plant extract prepared according to Examples 1 to 4 significantly increased collagen biosynthesis in a concentration-dependent manner.

In the case of the treatment concentration of 100 ppm, it was the highest in the first embodiment, followed by the second embodiment and the third embodiment. The ethanol plant - extracted ice plant composition was significantly higher than the other compositions, and the lowest concentration was found in the ice plant extract fermented by Bacillus subtilis.

3-2. Elastase  Measurement of inhibitory activity

The amount of p-nitroanilide produced from the substrate at 37 ° C for 20 minutes was measured at 405 nm using N-succinyl- (L-Ala) 3-pnitroanilide as a substrate.

0.5 mL of porcine pancreas elastase (2.5 U / mL) dissolved in 50 mM tris-HCl buffer (pH 8.6) was added to the test tube, and 50 mM N-succinyl- (L-Ala) 3-p-nitroanilide (0.5 mg / mL) dissolved in Tris-HCl buffer (pH 8.6) was added and reacted for 20 minutes. The measurement results are shown in Table 5 same.

Elastase inhibitory activity showed the decrease of the absorbance of the sample solution added and the no-added solution, and the following equation (4) was used.

&Quot; (4) "

Elastase activity inhibition rate (%) = [1- (a / b)] 100

a: absorbance of the treated group

b: Absorbance of the control group

sample Treatment concentration
(ug / ml) Elastase activity inhibition (%) First Embodiment Example 2-1 Example 2-2 Example Example 2-3 Third Embodiment Fourth Embodiment Ice Plant Extract 50 52.6 15.2 32.5 22.4 29.8 22.6 100 75.6 32.1 51.7 34.8 41.5 46.5 500 81.4 52.4 66.8 62.7 85.5 72.5 1,000 95.5 63.6 80.6 88.6 91.6 87.9

As shown in Table 5, it was found that the ice plant extract prepared according to the first to fourth embodiments significantly inhibited the elastase activity in a concentration-dependent manner.

Among them, the treatment concentration was the highest in the first embodiment when the treatment concentration was 1,000 ug / ml, followed by the third and the second to third embodiments.

The ethanol plant - extracted ice plant composition was significantly higher than the other compositions, and the lowest concentration was found in the ice plant extract fermented by Bacillus subtilis.

4. Antibacterial effect

The antimicrobial activity was measured using a paper disc method. Each strain cultured on a plate medium was plated and incubated for 24 hours in a liquid medium (50 mL), and then 0.5 mL of the bacterial liquid was inoculated in 50 mL of a liquid medium. The cells were inoculated at a density of about 1 × 10 ⁷ cells per plate medium and homogeneously smoothed with a sterilized cotton swab.

The diameter of the clear zone (mm) around the disc was measured by placing a sterilized filter paper disc (8 mm) on a solid plate medium and incubating the sample at a concentration of 0.05 mL / disc for each concentration at 37 ° C for 24 hours.

The antimicrobial effects of the ice plant extracts are shown in Table 6 below.

Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Propinoibacterium acnes were used to investigate the antimicrobial effect.

As shown in Table 6, it was found that the ice plant extract prepared according to Examples 1 to 4 increased the antimicrobial activity in a concentration-dependent manner.

In the case of the treatment concentration of 1 mg / disc, the growth inhibition rate of the four strains was the highest in the first example, followed by the second, third and the second examples.

In particular, it is believed that the antimicrobial activity of the fermented ice plant composition is high, and that the antimicrobial substance fermented in lactic acid bacteria or yeast, organic acids, proteolytic enzymes, etc. The ice plant extract is most effective in the growth of Staphylococcus aures And inhibited the growth of Propinoibacterium acnes.

It is considered that the effect of the ice plant extract is different according to the characteristics of each strain, and therefore it is considered that the result of this experiment should be well prepared when manufacturing the cosmetic composition after boiling.

5. Moisturizing effect

The ice plant extracts of the first to fourth embodiments were formulated into packs to examine the moisturizing effect.

The formulation of the pack is as shown in Table 7 below. The clinical moisturizing effect of the packaged cosmetic composition prepared by dividing 20 healthy women into 7 groups of 5 each was measured as follows.

The packs were used twice a week for 4 weeks. The moisturizing effect was measured using a Corneometer CM820PC (Courage + Khazaka, Germany) before, 3 and 5 weeks after the test. Table 9 shows the results of the confirmation of the performance of the product in order to check whether dermatitis occurs.

ingredient content(%) Ice plant extract First to fourth embodiments 1.0 Sodium alginate oligosaccharide 1.0 Ethyl alcohol 0.5 Stearic acid 0.5 glycerin 5.0 Propylene glycol 4.0 Polyoxide 0.05 Beta Glucan 1.0 antiseptic a very small amount Spices a very small amount Purified water Balance system 100

sample Example Moisturizing effect Before the test After 3 weeks After 5 weeks Ice plant
extract One 33.4 38.5 42.6 2-1 32.5 33.4 35.1 2-2 33.5 36.7 41.5 2-3 31.6 36.2 37.9 3 34.5 37.5 40.8 4 33.0 36.6 40.5

As shown in the above Table 8, when the pack was prepared using the ice plant extract and the moisturizing effect before, 2 weeks, and 4 weeks after the test was compared, the moisturizing effect was the highest in the first embodiment, Example 2-1 showed the lowest moisturizing effect.

It was found that the extract of ice plant showed remarkable moisturizing effect of the skin after use compared with before use.

sample Example Number of subjects who responded after 12 hours (persons) Subjects who responded after 24 hours (persons) 0 One 2 3 4 5 0 One 2 3 4 5 Ice Plant Extract One 20 0 0 0 0 0 0 0 0 0 0 0 2-1 20 0 0 0 0 0 0 0 0 0 0 0 2-2 20 0 0 0 0 0 0 0 0 0 0 0 2-3 20 0 0 0 0 0 0 0 0 0 0 0 3 20 0 0 0 0 0 0 0 0 0 0 0 4 20 0 0 0 0 0 0 0 0 0 0 0

As shown in Table 9, the composition containing the ice plant extract was found to be a safe substance in the human body since it belongs to the non-stimulated range.

As a result, the method of extracting an effective component of an ice plant for a cosmetic composition according to embodiments of the present invention is characterized in that, through the above-described technical constructions, an effective component of an ice plant having antioxidant activity, whitening activity, anti-inflammatory activity, antibacterial activity, Can be easily and easily extracted. The extracted ice plant extract according to the embodiments of the present invention can prevent or ameliorate skin diseases caused by oxidative stress and inflammation, and is useful as a composition for external use for skin and a cosmetic composition Can be used.

In addition, the extracted ice plant extract according to the embodiments of the present invention promotes differentiation of keratinocyte cells, restores skin barrier function, eliminates the pulling and roughness of the skin, and increases the elasticity, There is an excellent effect that can be usefully used.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is clearly understood that the same is by way of illustration and example only and is not to be taken by way of limitation in the present invention. Various changes and modifications may be made by those skilled in the art.

Claims (5)

Drying the leaves and stems of the washed ice plant;
Pulverizing the dried ice plant to a predetermined particle size;
Immersing the powdered ice plant in an aqueous ethanol solution for a predetermined period of time, and extracting an effective component of the ice plant;
Concentrating the extracted active ingredients of the ice plant under reduced pressure to remove the ethanol; And
A method for extracting an effective component of an ice plant for a cosmetic composition, which comprises lyophilizing an active ingredient of the ice plant concentrated under reduced pressure.
Drying the leaves and stems of the washed ice plant;
Pulverizing the dried ice plant to a predetermined particle size;
Adding a strain to the powdered ice plant, and then culturing to prepare an ice plant culture liquid;
Centrifuging and filtering the culture solution to prepare a filtrate from which the strain has been removed; And
Aging the filtrate for a predetermined period of time, and then re-filtering the mixture to obtain an extract containing the ice plant active ingredient, thereby extracting an effective component of an ice plant for a cosmetic composition.
3. The method of claim 2,
The strain may be selected from the group consisting of Bacillus subtilis , Saccharomyces cerevisiae, wherein the extract is one selected from the group consisting of Lactobacillus acidophilus , Lactobacillus acidophilus , Lactobacillus acidophilus , and Lactobacillus acidophilus .
Drying the leaves and stems of the washed ice plant;
Pulverizing the dried ice plant to a predetermined particle size;
Pouring the powdered ice plant; And
A method for extracting an effective component of an ice plant for a cosmetic composition, comprising the step of shaving the steamed ice plant.
Drying the leaves and stems of the washed ice plant;
Pulverizing the dried ice plant to a predetermined particle size; And
A method for extracting an effective component of an ice plant for a cosmetic composition, comprising the step of extracting an effective component of an ice plant using the supercritical extraction method.

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