KR20170051077A - Composition comprising extract from liriope platyphylla and natural extract - Google Patents

Abstract

The present invention relates to a composition comprising at least one natural extract, as an active ingredient, selected from a group consisting of: a Liriope muscari extract; Platycladus orientalis; Vitex trifolia; Cyperus rotundus; Chrysanthemum indicum; Morus alba; Ligusticum striatum; Angelica dahurica; Asiasarum heterotropoides; Thuja orientalis; Eclipta prostrata; Zingiber officinale; and Panax ginseng. The present invention provides the composition having excellent effects of hair loss and scalp improvement, and having excellent anti-dandruff, anti-inflammatory, antimicrobial, and anti-sebum effects.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition containing an extract of Lycoris chejuensis and a natural extract as an effective ingredient.

The present invention relates to a composition containing marshmallow extract and natural extract as an active ingredient.

The causes of hair loss have been discussed in male or female organs such as hair follicles, hair follicle blood flow, dysplasia, scalp irregularity due to excessive sebum secretion, genetic factors, and aging. However, the cause of hair loss has not yet been clarified yet. Recently, more and more people are suffering from alopecia, and the age group is lowering.

The human body has more than 1.3 million hairs and more than 100,000 hairs. These hairs go through three cycles of anagen, catagen, and telogen, and each hairs is operated in an independent cycle. Hair loss refers to a state in which hair or hair is less or no more than normal due to any cause, and it is distinguished by general hair loss phenomenon (telogen effluvium) and abnormal hair loss phenomenon (anagen effluvium) The latter is generally referred to as hair loss. Alopecia Areata, Androgenetic Alopecia, Telogen Effluvium, Traumatic, Trichotillomania, Pressure, and Alopecia are the most common forms of hair loss (or hair loss disease) (Alopecia syphiltiac), Alopecia seborrhecia, symptomatic, scarring, congenital alopecia, and the like.

As a conventional method for treating alopecia, there is a preparation based on female hormone in relation to hormonal theory, but its use has been discontinued due to reports of skin inflammation and adverse effects caused by administration of hormone. Minoxidil of US-based company and trichosaccharide of Crinos Co., Ltd. are used as hair growth promoters, but there is a problem of lack of clear effect and side effects. In addition, finasteride, developed by Merck, is known to inhibit the activity of 5-α-reductase, an enzyme that acts on the male hormone testosterone metabolism in hair follicles, but can cause depression This also has the problem that long-term treatment is necessary. In addition, there are Crescina and Rogaine, but only when the hair follicle is alive on the normal scalp before the hair loss disease is fully progressed. Thus, various conventional hair growth promoting methods have generally been attempted to promote hair growth with the aim of promoting blood circulation and nourishment of hair follicles, but the toxicity and side effects are also severe and the effect thereof is insufficient at present .

However, there is no known specific composition that can exhibit visible effects on hair loss, and a composition capable of exhibiting other efficacy besides improving hair loss is also known There is no known concrete yet.

Disclosed is a composition which is superior to conventional marshmallow extract composition and has excellent anti-hair loss and scalp improving effect, excellent antidandruff effect, excellent anti-inflammatory effect, excellent antimicrobial effect and excellent anti-sebum effect.

The present invention relates to an extract of marshmallow; And at least one natural extract selected from the group consisting of white porcelain, porcelain, coriander, gingko, mulberry, cinnabar, white porcelain, sezin, papaya, lavender, ginger and ginseng as an active ingredient.

The cosmetic composition according to the present invention is excellent in anti-dandruff effect, anti-dandruff effect, anti-inflammatory effect, antimicrobial effect and anti-dandruff effect by containing natural extract of peculiar constitution, And excellent anti-sebum effect.

Hereinafter, the composition according to the present invention will be described in detail.

Macmun dong is a plant of lilies and scientific name is Ophiopogon Japonicus or Liriope platyphylla . Flavonoids such as Staphyloidal saponin, Ophioponanone CF, and Ophiopogonone C, such as Ophiopogonon AE and Spicatoside AB, are known to be contained. It is reported that there is a difference in the composition depending on the method.

Chrysanthemum is a plant of Asteraceae and its scientific name is Chrysanthemum indicum . The ingredients that are included include phenolic compounds such as linarin and quercetine, and flavonoid compounds such as Cynarin.

The mongolian is a plant of marcheonopsis and the scientific name is Vitex trifolia or Vitex rotundifolia . Flavonoids such as Vitexcarpin, Artemetin, and Hesperidin are known to be included.

Ginseng is a plant of Araliaceae. It is Panax ginseng . Saponins such as Ginsenoside Rb1-2 are known to be included.

Hansikcho is a plant of Asteraceae and its scientific name is Eclipta prostrata . Examples of ingredients that may be included include flavonoids such as saponin such as Eclalbasaponin I-IV and Wedelolactone.

Pterygium is a plant of the genus Thuja Orientalis leaves are known to have excellent antibacterial and anti-inflammatory properties. The herbalist is Cyperus rotundus , the scientific name of the herbaceous plant, and the traditional medicinal herb has the effect of relieving pain and helping the gingiva.

White porcine leaves are the leaves of the cinnabar and plant. Its scientific name is Thuja orientalis .

Morus alba is a plant of Morus alba and its scientific name is Morus It is alba . Containing ingredients include Mrusin, Kuwanon ER and other flavonoids.

Ginger is a plant of ginger and the scientific name is Zingiber officinale . Phenolic compounds such as gingerone and shoagaol are known to be included.

The cinnabar is the plant of the persimmon, and the scientific name is Cnidium It is officinale ..

In one embodiment of the present invention, the content of the extract is from 0.01% by weight to 10% by weight based on the total weight of the composition. When the content is less than 0.01% by weight, hair loss prevention and scalp improvement are hardly observed, and if it exceeds 10% by weight, the formulation may be affected.

In one embodiment of the present invention, the content of the natural extract is 0.01 wt% to 10 wt% with respect to the total weight of the composition.

In one embodiment of the present invention, the composition is selected from the group consisting of McMundong; And water is added to at least one selected from the group consisting of white porcelain, porcelain porcelain, porcelain porcelain, porcelain, bamboo, cinnabar, white porcelain, And contains the obtained extract.

In one embodiment of the present invention, the composition is selected from the group consisting of McMundong; And water is added to at least one selected from the group consisting of white porcelain, porcelain, porcelain, Korean red gruel, white gruel, white gruel, white gruel, And contains the obtained extract.

The extracts contained in the composition according to the present invention may be those obtained by extracting and isolating from natural substances by a commonly used method for extracting an active ingredient from a plant, and the 'extract' , And may include both hot-water extract, polar solvent-soluble extract, and non-polar solvent-soluble extract.

Suitable solvents for preparing the extracts include any of those accepted in the art, and water or an organic solvent can be used. Examples of the solvent include alcohols having 1 to 4 carbon atoms, acetone, ether, and the like, including purified water, methanol, ethanol, propanol, isopropanol, butanol, Various solvents such as benzene, chloroform, ethyl acetate, methylene chloride, hexane and cyclohexane may be used alone or in combination. But is not limited to.

As the extraction method, any one or more of methods such as hot water extraction, cold extraction, reflux cooling extraction, solvent extraction, steam distillation, ultrasonic extraction, leaching, and pressing can be selected and used. In addition, the desired extract may be further subjected to a conventional fractionation process or may be purified using a conventional purification method. As a method of producing the extract of the present invention, conventionally known methods can also be used.

For example, the extract contained in the composition of the present invention can be prepared into a powdery state by an additional process such as vacuum distillation, freeze-drying, spray-drying, or the like, with the primary extract extracted by the hot water extraction or solvent extraction method described above. Further, the primary extract can be further purified by using various chromatographies such as silica gel column chromatography, thin layer chromatography, high performance liquid chromatography and the like, You can get it.

Therefore, in the present invention, the extract includes all the extracts, fractions, and purified products obtained in each step of extraction, fractionation or purification, their diluted solutions, concentrates or dried products, and the extracts may be prepared by mixing separate extracts of the respective herbals Or may be prepared by mixing together herbal medicines and then extracting them all at once. In addition, the extract of the present invention can be dissolved in 1,3-butyleneglycol and incorporated into the composition.

In one embodiment of the present invention, the composition is prepared by adding ethyl acetate to the extract obtained by the hot water extraction or the low-temperature extraction, and extracting the obtained ethyl acetate fraction into ethyl acetate fractions do.

In one embodiment of the present invention, the composition contains an extract obtained by fractionating ethyl acetate into a layer separated by separating butanol into a layer not separated by the ethyl acetate fraction.

In one embodiment of the present invention, the composition contains the extract obtained by the hot water extraction or the ethanol extract obtained by the low temperature extraction.

The cosmetic composition according to the present invention is excellent in the protection of the papilla cells, and is useful for preventing hair loss or improving scalp.

In addition, the cosmetic composition according to the present invention is for antioxidation with excellent free radical inhibition and active oxygen inhibition.

In addition, the cosmetic composition according to the present invention is an anti-dandruff agent having an excellent inhibitory effect on dandruff causative bacteria.

In addition, the cosmetic composition according to the present invention is an antibacterial composition having an excellent inhibitory effect on bacteria.

In addition, the cosmetic composition according to the present invention is a sebum-suppressing agent excellent in the effect of inhibiting sebum production.

In one embodiment of the present invention, the composition is a cosmetic composition. The composition may be further added with other ingredients as cosmetics to the extent that the composition is externally soluble in the skin.

In one embodiment of the present invention, the composition is a food composition. The composition may be further added with other ingredients within the range of ingredients that the human is allowed to ingest.

Further, the cosmetic composition according to the present invention is an antioxidant cosmetic composition having excellent free radical inhibition and active oxygen inhibition effect.

Further, the cosmetic composition according to the present invention is a cosmetic composition for improving or preventing wrinkles, which is excellent in an effect of inhibiting elastase, excellent in collagen synthesis activity, and excellent in inhibiting effect on MMP-1.

In addition, the cosmetic composition according to the present invention is an anti-inflammatory cosmetic composition having excellent NO inhibition.

In addition, the cosmetic composition according to the present invention is an anti-aging cosmetic composition having excellent skin moisturizing effect, antioxidant effect, skin wrinkle improvement or preventive effect and anti-inflammatory effect.

Hereinafter, the present invention will be described in more detail by way of examples. However, these embodiments are intended to illustrate the present invention only, and the scope of the present invention is not limited by these embodiments.

< Example  1> Hot water extraction  Preparation of the extracted composition

100 g of each of purified water was added to 100 g of purified water, and then 95 .mu.l of purified water was added to each 100 g of ginseng root, white porcine, white porcelain, giant porcelain, To 100 &lt; [deg.] &Gt; C for 10 to 15 hours under reflux. After completion of the extraction, the raw material was removed, and the extract was concentrated using a rotary-type cannula concentrator and lyophilized to obtain a solid-state extract.

< Example  2> Preparation of composition extracted by low temperature extraction

100 g each of purified water is added to 1 L to 2 L of purified water, and the mixture is stirred at room temperature of 20 to 25 ° C for 1 to 3 hours. Lt; / RTI &gt; for 1 week to elute the active ingredient. The extracted extract was concentrated as a rotary type goat cancer concentrator and lyophilized to obtain a solid-state extract.

< Example  3> Preparation of the composition extracted with ethyl acetate fraction

To the composition of Example 1, 1 L to 2 L of purified water was added, and the resulting precipitate was removed by filtration. Ethyl acetate was added thereto and mixed well. The mixture was allowed to stand to separate the layers. The upper layer was separated and concentrated using a rotary vacuum concentrator And lyophilized to obtain a solid-state extract.

< Example  4> Butanol  Preparation of compositions extracted with fractions

1 L to 2 L of butanol was added to the lower layer of the composition of Example 3, and the mixture was allowed to stand. After separating the layers, the upper layer was separated, concentrated using a rotary vacuum concentrator, and lyophilized to obtain a solid extract.

< Example  5> Preparation of composition extracted with polysaccharide fraction

The composition of Example 1 was concentrated using a rotary type carcinogen concentrator, ethanol was added to the concentrate, and the resulting precipitate was separated and vacuum-dried to obtain a solid-state extract.

< Experimental Example  1> Antioxidant efficacy measurement

Free radical scavenging test and reactive oxygen scavenging test were carried out to confirm the antioxidant effect. The samples used in the tests were the samples obtained in Examples 1 to 5, and ascorbic acid (L-ascorbic acid) having excellent antioxidative effect was used as a positive control in order to compare the antioxidative effect.

The free radical scavenging test utilizes the fact that the absorbance of stable DPPH exhibits the maximum absorbance at 540nm, and as the free radical DPPH is erased by the sample, it becomes clear color from purple to absorbance at the 540nm wavelength as the free radical scavenging rate increases And the test method was applied by the test method.

1 ml of 0.1 mM DPPH (2,2-diphenyl-1-picryl-hydrazyl radical, Sigma) solution was diluted to the appropriate concentration in methanol and mixed with 1 ml of the composition obtained in the above Examples 1 to 5, , And the absorbance was measured at a wavelength of 540 nm using a microplate reader (UVT-06685, Thermo max, USA)

In the free radical scavenging test, 1 ml of DPPH and 1 ml of methanol were added to the control, and 1 ml of methanol and 1 ml of the sample were added to obtain the respective color correction values for the sample and the control. The free radical scavenging ratio was calculated using the following equation (1). IC ₅₀ is the concentration of the sample required to eliminate free radicals by 50%, and the smaller the value, the higher the antioxidant activity.

[Equation 1]

Free radical scavenging rate (%) = 100 - ((absorbance of sample / absorbance of control) x 100)

The active oxygen scavenging test was carried out by measuring the absorbance change by oxidation of nitroblue tetrazolium (NBT) by reactive oxygen species using active oxygen generation by enzyme reaction of xanthine / xanthine oxidase It is possible to know the scavenging ability of the active oxygen.

2.4 ml of sodium carbonate (Na ₂ CO ₃ ), 0.1 ml of xanthine, 0.1 ml of ethylenediamine tetraacetic acid (EDTA), 0.1 ml of bovine serum albumin (BSA, Sigma) Add 0.1 ml of the sample and vortex using a vortex mixer (Type 37600 Mixer, USA) for 10 minutes at 25 ° C. Add 0.1 ml of xanthine oxidase, incubate at 25 ° C for 20 minutes, The reaction was stopped by adding 6 mM CuCl ₂ , and the absorbance was measured at a wavelength of 540 nm using a microplate reader

The control group in the active oxygen scavenging activity test was prepared by adding the third distilled water instead of the sample solution and measuring by the same method and adding the third distilled water instead of the residual enzyme solution to obtain the respective color correction values for the extracted sample and the control group .

The active oxygen scavenging rate was calculated using the following equation (2). IC ₅₀ is the concentration of the sample required to remove 50% of the active oxygen, and the smaller the value, the higher the antioxidant activity.

&Quot; (2) &quot;

(%) = 100 - ((absorbance of sample / absorbance of control) x 100)

The free radical scavenging ratio and the active oxygen scavenging ratio of the compositions of Examples 1 to 5 were measured, and L-ascorbic acid was used as a control group, and the measurement results are shown in Table 1 below.

Name of sample Free radical scavenging rate IC ₅₀ (ug / mL) The active oxygen scavenging rate IC ₅₀ (ug / mL) L-ascorbic acid 4.85 6.02 Example 1 8.87 7.26 Example 2 5.56 6.88 Example 3 6.05 8.45 Example 4 5.09 6.50 Example 5 5.88 7.11

As shown in Table 1, the antioxidative activities of the low-temperature extracts were superior to the hot-water extracts in Examples 1 to 5 according to the present invention. The antioxidant activities of the fractions were determined by the butanol fraction, Polysaccharide fractions, and ethyl acetate fractions.

< Experimental Example  2> Dermal papilla cells  Measurement of protection effect

The dermal papilla cells are the cells located at the base of the hair follicle. They supply oxygen and nutrients to the cells constituting the hair follicle, and control hair growth and follicle cycle regulation. Therefore, when the growth of the dermal papilla cells is promoted, the hair becomes healthy, hair growth is promoted, and hair loss can be prevented. The growth of hair progresses as the epithelial cells surrounding the dermal papilla divide and form the hair shaft, so the dermal papilla cells play an important role in controlling the division of the epithelial cells. In addition, in male alopecia, the region where the male hormone acts on the hair follicle also plays a very important role in hair growth.

Hair requires vigorous cell division for sustained growth and is thus an organ with a high level of oxidative stress. Thus, a composition capable of protecting hair dermal papilla cells from oxidative stress is anticipated to be effective in preventing hair loss, and evaluated whether the composition could protect hair dermal dermal cells from oxidative stress.

The dermal papilla cells were inoculated in 96 wells for 5, 000 hours, and cultured for 24 hours. The cells were treated with DMEM medium without fetal bovine serum for 48 hours. The cells were treated with 1 mM of hydrogen peroxide for 2.5 hours, washed, and cultured in DMEM medium without fetal bovine serum for 24 hours. The survival rate was measured by WST-1 and the results are shown in Table 2 below.

Cell survival rate (%) The negative control (H ₂ O ₂ No treatment) 100 Positive control group 23.15 Example 1 500ppm 57.47 250ppm 49.85 Example 2 500ppm 71.51 250ppm 51.65 Example 3 500ppm 39.56 250ppm 21.01 Example 4 500ppm 63.55 250ppm 50.24 Example 5 500ppm 60.15 250ppm 41.02

As shown in Table 2, Examples 1 to 5 according to the present invention were found to be excellent in the protection of papillary cells from oxidative stress, and in particular, Example 4 was found to be excellent in protecting papillary cells from oxidative stress.

< Experimental Example  3> Measurement of dandruff formation inhibitory activity

Dandruff is a keratinous mass formed by the dead cells or scraped cells of the scalp caused by the metabolism of the human body, which are dried together with the secretions of scalp sebaceous glands. Dandruff gradually progresses to dermatitis, usually with itching and inflammation, with a clear boundary with normal skin. Thus, the substance inhibiting the production of dandruff was anticipated to effectively work to prevent hair loss and improve the scalp, so that the dandruff inhibitory effect was evaluated to evaluate whether or not the composition inhibits dandruff formation.

In order to confirm the dandruff formation inhibitory effect of the composition, the inhibitory effect on dandruff causative bacteria Pityrosporum ovlae (ATCC 12087) was measured. The composition was diluted stepwise in the broth using the broth dilution method, the microorganism was inoculated in the diluted solution, and cultured in a 35 ° C incubator for 48 hours to measure the minimum inhibitory concentration (MIC) The results are shown in Table 3 below.

Minimum inhibitory concentration (MIC) Example 1 2.10 Example 2 1.95 Example 3 3.15 Example 4 1.71 Example 5 2.52

As shown in Table 3, Examples 1 to 5 according to the present invention showed excellent dandruff formation inhibitory activity.

< Experimental Example  4> Anti-inflammatory efficacy measurement

To confirm the anti-inflammatory effect of the composition, rat macrophages RAW264.7 cells were purchased from ATCC. (Dulbecco's modified Eagle's medium / Nutrient Mixture Ham's F12), FBS (fetal bovine serum), L-glutamine, penicillin-streptomycin and insulin used for cell culture were purchased from Gibco / BRL USA). The RAW 264.7 cells were cultured in DMEM / F12 medium supplemented with 10% FBS, 1% penicillin streptomycin 1% L-glutamine and 20 μg / ml insulin, and incubated at 37 ° C in a humidified CO 2 incubator (5% CO2 / 95% air). After the cells were cultured to about 80% of the culture dish, cells were washed with PBS (pH 7.4), washed, and subcultured by treatment with 0.25% trypsin and 2.56 mmol / L EDTA

The composition and LPS were added together and cultured for 24 hours. After 24 hours of culture, the culture was collected and the amount of NO secreted was measured using a Griess reagent system (Promega, USA) . The method using the Griess reagent system was performed in accordance with the protocol provided by Promega, which is the manufacturer of the system. Sulfanilamide solution was added to the collected cell culture solution, NED solution was added, and microplate reader At 540 nm, and the results are shown in Table 4 below.

Name of sample Treatment concentration NO inhibition rate (%) Example 1 500ppm 60.12 250ppm 43.53 Example 2 500ppm 35.94 250ppm 27.84 Example 3 500ppm 39.65 250ppm 17.85 Example 4 500ppm 89.15 250ppm 76.66 Example 5 500ppm 45.11 250ppm 29.56

As shown in Table 4, Examples 1 to 5 according to the present invention were found to be excellent in anti-inflammatory activity, and in particular, it was confirmed that the anti-inflammatory activity of Example 4 was excellent.

< Experimental Example  6> Measurement of antimicrobial efficacy

The antimicrobial activity test for the composition was carried out by an agar serial dilution method and a paper disc method. Experimental strains received pre-sale from the Korea Institute of Biotechnology, staphylococci (Staphylococcus aureus, KCTC 6910), Pseudomonas aeruginosa as gram-negative Gram-positive bacteria (Pseudomonas aeruginosa, KCTC 1637), Escherichia coli (E. coli, KCTC 1039), a yeast Candida yeast (Candida albicans , KCTC 7965), and aspergillus species ( Aspergillus niger , KCTC 6910).

In the case of yeast, the bacteria were inoculated on a potato dextrose medium and pre-cultured at 25 ° C for 2 days. The filamentous fungus was cultured on a potato dextrose agar medium The bacteria were inoculated and cultured for 7 days in a 25 ° C incubator. Spores of filamentous fungi formed on the surface of the culture medium were recovered by using a smear bar and diluted in sterilized saline.

2 ml of each extract diluted to the appropriate concentration in 5% dimethylsulfoxide (DMSO) physiological saline solution was added to the sterilized Patriedish according to the sample and the experimental species, and the control group was diluted to an appropriate concentration in 5% physiological saline solution Add 2 ml of each sample, add 2 ml of 5% DMSO physiological saline solution to the control group, sterilize each of the Petri dishes, add 18 ml of a tryptic soy agar medium and a potato dextrose agar medium cooled to 48 ° C, Let it stand and solidify.

Each of the pre-cultured bacteria was applied to each petri dish at a final concentration of about 1 × 10 ⁶ CFU / ml in the case of bacteria, 1 × 10 ⁵ CFU / ml for yeast, about 1 Spread each petri dish at a bacterial concentration of X 10 ⁴ CFU / ml. Each petri dish was incubated at 37 ° C for 24 hours. Yeast was cultured at 25 ° C for 3 days. Molds were cultured in a 25 ° C incubator for 7 days, and colonies were observed in each compartment.

The minimum inhibitory concentration (MIC) of the untreated plate is defined as the minimum inhibitory concentration (MIC).

The results of the antibacterial test are shown in Table 5 below.

sample Minimum inhibitory concentration (MIC) S. aureus P. aeruginosa E. coli C. albicans A. niger Example 1 0.5 0.5 0.3 0.3 0.6 Example 2 0.6 0.7 1.1 1.0 0.5 Example 3 0.3 0.3 0.6 0.5 0.5 Example 4 0.2 0.3 0.2 0.2 0.1 Example 5 0.3 0.4 0.2 0.2 0.2

As shown in Table 5, it was confirmed that Examples 1 to 5 according to the present invention are excellent in antibacterial activity.

< Experimental Example  6> Sebum secretion  Measurement of inhibitory effect

The sebaceous gland cells were obtained from hair follicles in the dermal layer and the isolated sebaceous glands were treated with 0.25% Trypsin, 0.05% Ethylenediamine Tetraacetic Acid (EDTA) for 30 min. , And trypsin was inactivated with MEM (Eagle's minimum essential medium) containing 12% fetal bovine serum (FBS). Then, the medium was removed by centrifugation, and then KGM (keratinocyte growth medium, and cultured in a 60 mm culture dish. 2 x 10 ⁵ cells were plated per well using a 24-well plate for this experiment during subculture. After the cells were fixed for 24 hours, the cells were dispersed in dimethylsulfoxide (DMSO) to a final concentration of 0.1% (w / v) and incubated for 72 hours. The amount of sebum produced was extracted and measured by methanol: chloroform extraction method to compare the amount of sebum produced per 10 ⁴ cells. The extraction method of sebum was performed by the method of Zoboulis et al. (J. Invest. Dermatol., 1991; 24, 69-83), and the extraction method of methanol: chloroform, ultrasonic disruption, , And nitrogen evaporation. The results of this experiment are shown in Table 6.

% Inhibition of sebum production (%) Example 1 38 Example 2 22 Example 3 31 Example 4 48 Example 5 41

As shown in Table 6, it was confirmed that Examples 1 to 5 according to the present invention are excellent in sebum production inhibiting activity.

< Experimental Example  7> Protein content analysis

A bicinchoninic acid (BCA) assay was performed to determine the protein content of the composition. The BCA assay uses a property that proteins can reduce copper ions (Cu ^{2 +} , Cu ^{1 +} ). Copper ions reduced by the protein react with BCA solution to form a violet compound (Cu ⁺ / BCA chromophore) This violet compound shows the maximum absorbance at a specific wavelength of 562 nm. Therefore, the amount of protein was measured by increasing the absorbance at 562 nm as the amount of protein was increased. The experimental method is as follows.

First, 150 μl of the standard solution or diluted sample was added to each well of 96 microplate using a micro BCA protein measurement reagent kit (Pierce, Cat. No. 23227), and then BCA (bicinchoninic acid) kit WR (Working Reagent) reagent And mixed vigorously for 30 seconds. Then, the reaction was carried out at 37 ° C for 2 hours, and a 540 nm absorbance was measured with a microplate reader (UVT-06685, Thermo max, USA) to prepare a calibration curve and quantify the protein. The results are as follows.

Name of sample Total protein content (%) Example 1 6.25 Example 2 5.10 Example 3 3.15 Example 4 7.51 Example 5 7.36

< Experimental Example  8> Analysis of constituent amino acids

0.2 g of the composition was hydrolyzed in 20 ml of 6N HCl at 130 DEG C for 24 hours to break the peptide bond of the protein and make it a free amino acid. Then, free amino acids were separated by using OPA (o-phthalaldehyde, Agilent) and FMOC (9-fluorenylmethylchloroformate) derivatives to form fluorescence isoindole, followed by high performance liquid chromatography (Agilent Technologies 1200 Series HPLC, Agilent, USA (FLD, Agilent Technologies, USA). The constituent amino acids of the 250 pM composition were identified as standards. The contents are shown in Table 8 below.

Amino
acid
(ppm) Asp Glu Ser His Gly The Arg Ala Tyr Cys Val Pro room
city
Yes One 2.21 0.85 0.71 1.21 - - 1.27 0.68 0.32 1.64 1.10 1.92 2 - 1.15 0.23 - 0.21 0.08 0.56 0.25 - 0.98 0.05 - 3 2.73 0.29 0.89 0.44 0.10 0.02 0.24 1.13 - - 0.66 - 4 0.35 - - 1.02 0.34 - - - 0.10 1.22 0.25 0.86 5 0.71 0.16 0.15 0.89 0.18 - 1.02 0.60 0.52 0.56 1.12 1.56

&Lt; Formulation Example of Formulation & Formulation Example  1 to 3 and Comparative Formulation Example  Preparation of 1 to 3

In order to check the feeling of use according to the formulation of the composition according to the present invention, hair loss preventing hair shampoo composition (Formulation Example 1) according to the composition of the following Table 9, hair loss preventing hair conditioner composition (Formulation Example 2) And an anti-hair restoration hair tonic composition (Formulation Example 3) according to the composition shown in Table 11 below.

In order to compare with the formulation according to the present invention,

General hair loss preventing hair conditioner composition (Comparative Formulation Example 2) according to the composition of the following Table 13 and the general hair loss preventing hair tonic composition (Comparative Formulation Example 3) according to the composition of the following Table 14 .

number Raw material Content (% by weight) One Polyquaternium-10 0.2 2 Sodium lauryl sulfate 15 3 Sodium laureth sulfate 45 4 Glycollosteralate 2 5 Cetearyl alcohol 0.5 6 Lauryldiethylamide 2.5 7 The composition of Example 4 1.0 8 Phenoxyethanol 0.7 9 Purified water 33.1

number Raw material Content (% by weight) One Cetyl alcohol 3.0 2 Stearyl alcohol 3.0 3 Propylene glycol 2.0 4 Cetyltrimethylammonium chloride 1.0 5 Polydimethylsiloxane 1.0 6 The composition of Example 4 1.0 7 Mineral oil 0.5 8 Citric acid 0.2 9 Phenoxyethanol 0.7 10 Purified water 87.6

number Raw material Content (% by weight) One menthol 0.3 2 ethanol 30 3 Salicylic acid 0.1 4 Propylene glycol 0.25 5 Triethanolamine 0.2 6 Panthenol 0.1 7 Phenoxyethanol 0.7 8 Purified water 68.35

number Raw material Content (% by weight) One Polyquaternium-10 0.2 2 Sodium lauryl sulfate 15 3 Sodium laureth sulfate 45 4 Glycollosteralate 2 5 Cetearyl alcohol 0.5 6 Lauryldiethylamide 2.5 7 Purified water 34.8

number Raw material Content (% by weight) One Cetyl alcohol 3.0 2 Stearyl alcohol 3.0 3 Propylene glycol 2.0 4 Cetyltrimethylammonium chloride 1.0 5 Polydimethylsiloxane 1.0 6 Mineral oil 0.5 7 Citric acid 0.2 8 Phenoxyethanol 0.7 9 Purified water 88.6

number Raw material Content (% by weight) One menthol 0.3 2 ethanol 30 3 Salicylic acid 0.1 4 Propylene glycol 0.25 5 Triethanolamine 0.2 6 Panthenol 0.1 7 Phenoxyethanol 0.7 8 Purified water 68.35

< Experimental Example  9> Shampoo formulations Feeling  Test

30 university students were divided into two groups and tested for their use for 4 weeks. The product was used more than 3 times a week. After using for 6 weeks, the questionnaire was conducted. The survey scale was as follows.

5 points: Very good / 4 points: Good / 3 points: Average / 2 points: No effect / 1 point: Worsening

The results of the questionnaire after Formulation Examples 1 to 3 according to the present invention and Comparative Formulation Examples 1 to 3 after the comparative usability test are shown in Tables 15 to 17 below.

As a result of the survey, it was confirmed that the composition according to the present invention tends to receive a higher score in the dosage form than the dosage form without the composition according to the present invention. Thus, it is confirmed that the composition according to the present invention has a clinical effect when the composition according to the present invention is added.

Survey item Formulation Example 1 Comparative Formulation Example 1 1. The itchiness of the scalp was reduced. 3.8 3.2 2. Dandruff decreased. 3.6 3.3 3. Scalp sebaceum decreased. 3.2 3.1 4. The thickness of the hair became thick. 3.0 2.9 5. I feel shine of hair after shampooing 4.0 3.5 6. Finger passes well during rinsing. 3.2 3.1 7. Hair is easy to care after drying and is soft. 3.4 3.1 8. After shampooing, the hair gets moist longer 3.7 3.5

Survey item Formulation Example 2 Comparative Formulation Example 2 1. The itchiness of the scalp was reduced. 3.3 3.2 2. Dandruff decreased. 3.4 3.1 3. Scalp sebaceum decreased. 3.2 3.2 4. The thickness of the hair became thick. 3.1 3.0 5. I feel gloss of hair after using conditioner 3.8 3.6 6. Finger passes well during rinsing. 3.6 3.1 7. Hair is easy to care after drying and is soft. 3.7 3.2 8. After the conditioner, the hair becomes moist longer 3.5 3.2

Survey item Formulation Example 3 Comparative Formulation Example 3 1. The itchiness of the scalp was reduced. 3.8 3.5 2. Dandruff decreased. 3.6 3.3 3. Scalp sebaceum decreased. 3.5 3.3 4. The thickness of the hair became thick. 3.5 3.2 5. I feel shine of hair after shampooing 3.7 3.5

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will readily appreciate that many modifications are possible in the exemplary embodiments without materially departing from the novel teachings and advantages of this invention. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (16)

Marshmallow extract; And
A composition comprising as an active ingredient one or more natural extracts selected from the group consisting of white porcine, mandarin, angular buds, red ginseng, alfalfa, cinnabar, white ginseng, sezin, papaya, lily of the valley, ginseng and ginseng. The method according to claim 1,
Wherein the content of the extract is from 0.01% by weight to 10% by weight based on the total weight of the composition. The method according to claim 1,
Wherein the content of said natural extract is from 0.01% to 10% by weight relative to the total weight of the composition. The method according to claim 1,
The composition is available from McMundong; And water is added to at least one selected from the group consisting of white porcelain, porcelain porcelain, porcelain porcelain, porcelain, bamboo, cinnabar, white porcelain, A composition containing the obtained extract. The method according to claim 1,
The composition is available from McMundong; And water is added to at least one selected from the group consisting of white porcelain, porcelain, porcelain, Korean red gruel, white gruel, white gruel, white gruel, A composition containing the obtained extract. 3. The method according to claim 1 or 2,
A composition comprising the extract obtained by extracting the hot water or the extract obtained by the low temperature extraction with ethyl acetate and separating into one layer which is separated into ethyl acetate. The method according to claim 6,
Wherein the extract is obtained by fractionating ethyl acetate by separating into one layer separated by adding butanol to the layer not separated by the ethyl acetate fraction. 3. The method according to claim 1 or 2,
A composition comprising the extract obtained by the hot water extraction or the ethanol extract obtained by the low temperature extraction. The method according to claim 1,
Wherein said composition is for preventing hair loss or improving scalp. The method according to claim 1,
Wherein said composition is for antioxidant. The method according to claim 1,
Wherein the composition is anti-dandruff. The method according to claim 1,
Wherein said composition is for antimicrobial use. The method according to claim 1,
Wherein said composition is anti-sebum. The method according to claim 1,
Wherein said composition is anti-sebum. The method according to claim 1,
Wherein the composition is a cosmetic composition. The method according to claim 1,
Wherein the composition is a food composition.