KR101176521B1 - Cosmetic compositions comprising mixed extracts of Zanthoxylum schinifolium, Pulsatilla koreana, Usnea barbata, Illicium verum Hooker fil., Echinacea and propolis - Google Patents

Abstract

The present invention relates to a cosmetic composition for skin improvement comprising, as an active ingredient, a mixed extract of Sancho, Baekduong, Moss, Octagonal Fennel, Echinacea and Propolis. The mixed extracts of Sancho, Baekduong, Moss, Octagonal Fennel, Echinacea and Propolis of the present invention not only have excellent antibacterial and anti-inflammatory effects, but also have much better effects than the respective natural extracts alone or less than 6 mixed extracts. There is. In addition, the cosmetic composition containing it is also excellent in atopic improvement.

Sancho, Baekduong, Moss, Octagonal Fennel, Echinacea, Propolis, Anti-inflammatory, Antibacterial, Atopy Improvement

Description

Cosmetic compositions comprising mixed extracts of vinegar, baekduong, moss, octagonal fennel, echinacea and propolis, as an active ingredient propolis}

The present invention relates to a cosmetic composition containing a mixed extract of Sancho, Baekduong, Moss, Octagonal Fennel, Echinacea and Propolis.

Many microorganisms exist in our living environment. Various microorganisms exist in the skin which is always exposed to the outside. The fungus found on the skin is divided into resident bacteria and transient bacteria. Intravenous bacteria are Gram-positive bacteria that, as permanent microcolonies on the surface and outermost layers of the skin, play an important beneficial role in preventing colonization of other more harmful bacteria and fungi. Transient bacteria are bacteria that are not part of the normal resident flora of the skin, but can attach when airborne contaminants touch the skin or when they are in physical contact with the contaminated material. Transient bacteria are typically divided into gram positive and gram negative. Gram-positive bacteria include pathogens such as Staphylococcus aureus , Streptococcus pyogenes , and Clostridium botulinum . Gram-negative bacteria Salmonella (Salmonella), in Sherry teeth coli (Escherichia coli), Klebsiella (Klebsiella), Haemophilus (Haemophilus), Pseudomonas ah rugi labor (Pseudomonas aeruginosa), Proteus (Proteus) and Shh Gela di Centennial Pathogens such as Shigella dysenteriae . Some of these skin flora may cause various skin diseases. In particular, Staphylococcus aureus , Streptococcus pyogenes , etc. are representative strains that can cause purulent infections, cellulitis or general inflammation on the skin.

Recently, due to environmental pollution and changes in diet, many skin diseases such as atopy are occurring. Atopy, atopia in Latin, is an odd word, meaning a constitution that is prone to hypersensitivity. Atopy causes a series of allergic symptoms in the skin and respiratory mucosa, eye mucosa and intestinal mucosa in individuals with atopic inducers. People with atopy-inducing factors often have urticaria reactions in skin tests for protein substances commonly encountered in the outside environment, such as house dust, mites, animal hair, food, pollen, and molds. Symptoms include itching, skin attacks, hives, angioedema, sneezing, runny nose, stuffy nose, conjunctival hyperemia and tearing. Allergic diseases caused by atopy-inducing factors include atopic dermatitis, allergic rhinitis, asthma, allergic conjunctivitis, allergic enteritis, and atopic urticaria. These diseases are caused by genetic predisposition, environment, and age of each patient. It may appear alone or in combination. Among them, atopic dermatitis is a representative allergic skin disease in people with atopic allergy.

Therefore, skin protection function from the external environment has emerged as an important problem for maintaining healthy skin. Cosmetics are also used to protect the skin from these external factors. Recently, there are a lot of attempts to use natural ingredients as inhibitors for substances that cause skin diseases.

On the other hand, Sancho (Zanthoxylum schinifolium) is a deciduous shrub belonging to the Unhyang family and is a fragrant plant with a characteristic scent from the leaves. It is also used as an herbal medicine. The components of the herb are reported as α-sansholl, γ-sanshool, hydroxy-γ-sanshool, hydroxy-α-sanshool, hydroxy-β-sanshool, xanthoxylin, mikanin, hyperin, isovanillic acid and quercetin. Studies on the functional properties of antioxidants and anti-thrombotic effects of root, stem and leaf extracts from wild vinegar, antimicrobial activity of wild vinegar extract against Vibrio parahaemolyticus , antimicrobial activity and chemical composition of vinegar and vinegar leaf have.

Baekduong ( Pulsatilla koreana ) is a perennial herb of the dicotyledonous plant of the genus Minaria, and is distributed in Korea, northeastern China, Ussuri river, and Heilong river. Nogocho, also called pale flowers. The chunks of white fluffy berries look like granny's white hairs, so they are named pale flowers. Root is used for antipyretic, astringent, analgesic, anti-inflammatory, hemostasis, sterilization, etc. or as a dysentery such as dysentery. Research on the antimicrobial activity has been reported on the identification and identification of antimicrobial activity and activity of Staphylococus aureus , MRSA, Proteus vulgaois, and antimicrobial and anti-inflammatory effects on periodontal pathogens .

Moss is the first plant to rise from the water to the ground. It has chloroplasts that photosynthesize and reproduce as holes. It is estimated that about 15,000 to 25,000 species of moss live on the planet, and about 5,000 of them are known to live in tropical rainforests. Moss absorbs rainwater and moisture from the leaves or stems, not from the roots, and when it dries, survives the dry environment in a dormant state and survives when wet again. Currently, research on extracting anti-aging substances from polarized moss and applying them to cosmetics is underway. Also, substances that are effective in whitening and improving wrinkles have been identified.

Octagonal fennel ( Ilicium verum Hooker fil.) Is the fruit of magnolia and evergreen. The name derives from the appearance of eight pods wrapped in hard shells, like stars. The fragrance is strong and the taste is spicy, so it is mainly used as a spice. Diuretic and food-boosting, abdominal bloating or nausea to reduce the action is. In China, it is used as a herbal medicine for the treatment of back pain, constipation and cystitis. It is also used to relieve colic or acute rheumatism.

Echinacea (Echinacea) is effective in the treatment and prevention of fine herbs strengthen the immune system, various kinds of inflammation is a very strong antibiotic as a hub sseudeon herbal when the North American Indians were bitten by poisonous snakes. In Europe, Ichinacea is primarily used to boost immune organs and to boost natural healing resistance against cold infections that infect the mucous membranes of the esophagus and larynx. Echinacea extract promotes the activity and development of macrophages, natural killer cells and T-cells, which are the main agents of the immune system, increasing the body's defense against infection by bacteria and viruses. It is supposed to be.

Propolis is a honeybee that collects resin-like substances extracted from various types of trees and herbs on the hind legs of honeybees for their survival and reproduction.These are mixed with their saliva and mixed with the enzymes in the saliva. General term for the composite of the various high molecular polymers produced by the reaction, its main component is a flavonoid. Flavonoids are anisozanins such as flavones, flavonones, flavonols, flavanols, flavanones and isoflavones and glycosides associated with sugars, hundreds of ansocyanins and glycosides associated with sugars, catechins as tannins And leucozancin. Propolis is B.C. Long ago 300 years ago, the ancient Egyptians used it effectively as an antiseptic to embalm mummies.These preservatives were used by moths, small rats, or snakes in the beehive, where they were killed by bees attack and then carried them out of the beehive. Failure to do so can be confirmed by natural observation that the propolis coated by the bees does not rot for years and the original remains intact. The non-preservative effects of propolis include anti-cancer, burn healing, eczema, atopic dermatitis and athlete's foot such as skin disease, hair loss inhibition, menstrual pain relief or elimination, constipation, skin care and wrinkle removal The ability to cure and prevent bacterial dental diseases such as dental caries, tooth decay, and oral infections has been reported. Based on this, the Ministry of Health and Welfare has recently defined health supplements in Korean Food Code 13-25. In addition, for flavonoids, the main ingredient of propolis, in 1969, Dr. Kabarkina's 5.6-fold increase in immunity effect was announced, and in 1973, he demonstrated stronger bactericidal activity than penicillin of Alexandria, Russia. Anti-inflammatory effect, the results of the anti-cancer effect clinical trials at the Japan Institute of Technology in 1994, activation of other systemic cells, hormonal stimulation, treatment of scurvy by the action of vitamin P, blood pressure lowering in hypertensive patients, balance of biological hormones The results of studies and clinical trials in countries around the world have been reported on their effectiveness.

To date, extracts of Sancho, Baekduong, Moss, Octagonal Fennel, Echinacea and Propolis have been known and used for various effects ranging from antibacterial, antioxidant and anti-inflammatory effects as well as whitening and wrinkle improvement. However, the antimicrobial, anti-inflammatory and atopy-improving effects of the mixed extracts extracted from Sancho, Baekduong, Moss, Octagonal Fennel, Echinacea and Propolis and the compositions containing them are increased compared to the use of each natural product alone. It has not been disclosed at all.

Therefore, the present inventors have conducted research on natural substances excellent in antimicrobial, anti-inflammatory and atopy improvement, and as a result, the mixed extracts of Sancho, Baekduong, moss, octagonal fennel, echinacea and propolis are particularly excellent for antibacterial, anti-inflammatory and atopy improvement. The present invention was completed by confirming. In particular, the six natural product mixed extracts exhibited antimicrobial and anti-inflammatory effects more than expected compared to less than six mixed extracts, and in the case of a cosmetic composition containing the mixed extract, the atopy treatment and improvement effect was also excellent.

Accordingly, it is an object of the present invention to provide a cosmetic composition excellent in antimicrobial, anti-inflammatory and atopy-improving effects containing mixed extracts of vinegar, baekduong, moss, octagonal fennel, echinacea and propolis.

Other objects and advantages of the present invention will become apparent from the following solutions, examples and claims.

The present invention relates to a cosmetic composition for skin improvement comprising, as an active ingredient, a mixed extract of Sancho, Baekduong, Moss, Octagonal Fennel, Echinacea and Propolis. The mixed extracts of Sancho, Baekduong, Moss, Octagonal Fennel, Echinacea and Propolis of the present invention not only have excellent antibacterial and anti-inflammatory effects, but also have much better effects than the respective natural extracts alone or less than 6 mixed extracts. There is. In addition, the cosmetic composition containing the excellent antimicrobial effect as well as atopy improvement effect.

In order to achieve the above object, the present invention provides a cosmetic composition for improving skin containing a mixed extract of Sancho, Baekduong, moss, octagonal fennel, Echinacea and propolis.

Hereinafter, the present invention will be described in detail.

In the present invention, the moss is a moss called Usnea barbata belonging to the genus (Urna) and grows on the bark of trees native to the northern hemisphere of Europe, Asia, and the United States. Yellow or greenish shrub lichen with long stems and disc-shaped attachments that resemble tangled threads. It can be seen from the Arctic to the tropics, where it is used as a food or feed for wildlife. In the past, it was used for the treatment of pertussis, Qatar, epilepsy, and also as an astringent, tonic, and diuretic. In addition, usnic acid is known for its antimicrobial activity.

The mixed extracts of sancho, baekduong, moss, octagonal fennel, echinacea and propolis, which are the active ingredients of the composition of the present invention, are conventional solvents according to conventional methods known in the art, that is, under conditions of ordinary temperature and pressure, It can be prepared using.

For example, the mixed extracts of the acid, white, moss, octagonal, echinacea and propolis of the present invention may be water, anhydrous or hydrous lower alcohols having 1 to 4 carbon atoms, acetone, ethyl acetate, butyl acetate and 1,3-butylene. It is preferably extracted using a solvent selected from the group consisting of glycols, more preferably water and alcohols, even more preferably water and ethanol. The ethanol is preferably 70% ethanol. Suitable amounts of the extraction solvent are 1-15 times, more preferably 1-5 times, most preferably 3 times the dry weight of acid, baekduong, moss, octagonal fennel, echinacea and propolis. The mixed extract may be prepared by any of the methods of mixing the extract of each natural product or mixing the natural product and then extracting (mixed extraction), preferably by mixing extraction. In addition, as extraction conditions, it is extracted for 1 hour to 5 days at a temperature of 4-100 ℃, preferably at a temperature of 90-100 ℃ for 1 hour to 5 hours, more preferably at a temperature of 95-100 ℃ Extraction is for 2 to 3 hours.

Specifically, the mixed extracts of Sancho, Baekduong, Moss, Octagonal Fennel, Echinacea and Propolis of the present invention are prepared by a process comprising the following steps:

(a) 1-5 times of water, dry anhydrous or hydrous lower alcohol in dry sancho, baekduong, moss, octagonal fennel, echinacea and propolis (dry weight ratio 1: 1: 1: 1: 1: 1) Adding a solvent selected from the group consisting of acetone, ethyl acetate, butyl acetate and 1,3-butylene glycol;

(b) extracting the acetic acid, baekduong, moss, octagonal fennel, echinacea and propolis to which the solvent of step (a) is added for 1 hour to 5 hours at 95-100 ° C;

(c) filtering the extract of step (b) and drying the filtrate at 65 ° C. on a rotary decompression evaporator.

In this case, when 1,3-butylene glycol is used as the extraction solvent, it is difficult to dry using a rotary depressurization evaporator, and thus, after performing the extraction step (b), the drying loss is adjusted to 1% (w / v).

On the other hand, the mixed extract of Sancho, Baekduong, Moss, Octagonal Fennel, Echinacea and Propolis of the present invention includes not only the extract by the aforementioned extraction method, but also the extract by the supercritical extraction method or the ultrasonic extraction method under reduced pressure and high temperature with carbon dioxide. do.

The decompression by carbon dioxide and the supercritical extraction by high temperature mean supercritical fluid extraction. Generally, the supercritical fluid is a liquid having a gas when the gas reaches a critical point under high temperature and high pressure. It is gaseous, chemically similar in polarity to nonpolar solvents, and because of its properties, supercritical fluids are used for extraction of fat-soluble substances (J. Chromatogr. A. 1998; 479: 200-205).

Carbon dioxide becomes a supercritical fluid with both liquid and gaseous properties as the pressure and temperature reach a critical point through the operation of a supercritical fluid device, resulting in increased solubility in fat-soluble solutes. When supercritical carbon dioxide passes through an extraction vessel containing a certain amount of sample, the fat-soluble substance contained in the sample is extracted from the supercritical carbon dioxide.

When the supernatant carbon dioxide containing a small amount of cosolvent is passed through the sample remaining in the extraction vessel after extracting the lipid-soluble substance, components that were not extracted by pure supercritical carbon dioxide can be extracted.

The supercritical fluid used in the supercritical extraction method of the present invention can effectively extract the active ingredient by using a supercritical carbon dioxide or a mixed fluid in which a co-solvent is added to carbon dioxide. The cosolvent may be used one or a mixture of two or more selected from the group consisting of chloroform, ethanol, methanol, water, ethyl acetate, hexane and diethyl ether.

Most of the extracted samples contain carbon dioxide. Since carbon dioxide is volatilized into air at room temperature, the extract obtained by the above method may be used as a cosmetic composition, and the cosolvent may be removed by a reduced pressure evaporator.

In addition, the ultrasonic extraction method is an extraction method using the energy generated by the ultrasonic vibration, the ultrasonic wave can destroy the insoluble solvent contained in the sample in the water-soluble solvent, the reactants located around due to the high local temperature generated Since the kinetic energy of the particles is increased, sufficient energy necessary for the reaction is obtained, and the effect of ultrasonic energy is induced to increase the pressure by increasing the mixing effect of the substance and the solvent contained in the sample, thereby increasing the extraction efficiency.

As the extraction solvent that can be used in the ultrasonic extraction method, one or a mixture of two or more selected from the group consisting of chloroform, ethanol, methanol, water, ethyl acetate, hexane and diethyl ether can be used.

The extracted sample is vacuum filtered to recover the filtrate, and then removed by a vacuum evaporator, the extract can be obtained through a conventional extract preparation method of freeze drying.

In addition, the mixed extract of the herbaceous acid, baekduong, moss, octagonal fennel, echinacea and propolis of the present invention also includes extracts which have undergone conventional purification and / or fermentation. Obtained by various additional purification methods, such as, for example, separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity). Fractions are also contemplated to be included in the mixed extracts of the herb, baekduong, moss, octagonal fennel, echinacea and propolis of the present invention. In addition, the fermented product obtained by fermentation for 1 to 7 days while maintaining pH 5 to 7 under temperature conditions of 20 to 40 ℃ as a bacterium selected from the group consisting of yeast, lactic acid bacteria, Bacillus bacteria and mixtures thereof, It is interpreted to be included in mixed extracts of baekduong, moss, octagonal fennel, echinacea and propolis.

The mixed extracts of the herbaceous acid, baekduong, moss, octagonal fennel, echinacea and propolis of the present invention may be prepared in powder form by additional processes such as distillation under reduced pressure and freeze drying or spray drying.

According to a more preferred embodiment of the present invention, the mixed extract of Sancho, Baekduong, Moss, Octagonal Fennel, Echinacea and Propolis of the present invention is antibacterial (Experimental Example 1, 2, 3), anti-inflammatory (Experimental Example 4), immune response Inhibitory effect (Experimental example 5) is excellent.

In a preferred embodiment of the present invention, the components included in the cosmetic composition is a variety of components to maintain the activity of the biological components in addition to the mixed extract of Sancho, Baekduong, moss, octagonal fennel, echinacea and propolis used as the active ingredient of the present invention Such as preservatives, osmotic agents, pH adjusting agents, buffers, stabilizers, aluminum hydroxide, aluminum phosphates, surfactants, liposomes, thickeners, and the like and components commonly used in cosmetic compositions such as antioxidants, stabilizers, solubilizers, Conventional adjuvants such as vitamins, pigments and flavors, and carriers and the like, may be included, but are not limited thereto.

Cosmetic compositions of the invention may be prepared in any formulation conventionally prepared in the art, including, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, It may be formulated as an oil, powder foundation, emulsion foundation, wax foundation, spray, and the like, but is not limited thereto. More specifically, it may be prepared in the form of a flexible lotion, nutrition lotion, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.

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

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular in the case of a spray, additionally chlorofluorohydrocarbon, propane Propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, and microcrystals are used as carrier components. Castle cellulose, aluminum metahydroxy, bentonite, agar and the like can be used.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, an isethionate, an imidazolinium derivative, a methyltaurate, a sarcosinate, a fatty acid amide ether. Sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

According to a preferred embodiment of the present invention, the content of the mixed extract of Sancho, Baekduong, Moss, Octagonal Fennel, Echinacea and Propolis is 0.0001-50.0% by weight, preferably 0.01-15.0% by weight, based on the total weight of the cosmetic composition. Most preferably, it is 0.1-10.0 weight%. When the content of the mixed extract of the acid, baekduong, moss, octagonal fennel, echinacea and propolis is less than 0.0001% by weight, there is a problem in showing antimicrobial, anti-inflammatory and atopy improving effects, synergistic effect when exceeding 50.0% by weight In addition to the unexpected, there is a problem in the stability of the formulation of the composition and may cause skin irritation. In addition, extracts of each natural product of Sancho, Baekduong, moss, octagonal fennel, echinacea and propolis or each natural product are mixed in a mass ratio of 1-3: 1-3: 1-3: 1-3: 1-3: 1-3. And most preferably mixed in equal mass ratios.

According to a specific embodiment of the present invention, the cosmetic composition containing a mixed extract of the acid, baekduong, moss, octagonal fennel, echinacea and propolis of the present invention is antibacterial (sterilization; Experimental Example 6) and atopy improvement (Experimental Example 7) The effect is excellent.

The present invention also provides a cosmetic method comprising applying a cosmetic composition containing the active ingredient of the present invention to human skin.

The cosmetic method of the present invention refers to all cosmetic methods for applying the cosmetic composition of the present invention to human skin. That is, all methods known in the art for applying the cosmetic composition to the skin belong to the cosmetic method of the present invention.

The cosmetic composition of the present invention may be used alone or in combination, or may be used by overlapping with other cosmetic compositions other than the present invention. In addition, the cosmetic composition with excellent skin protection effect according to the present invention can be used according to a conventional method of use, the number of times of use can be varied according to the user's skin condition or taste.

When the cosmetic composition of the present invention is a soap, surfactant-containing cleansing or surfactant-free cleansing formulation, it may be wiped off, peeled off or washed with water after application to the skin. As a specific example, the soap is liquid soap, powdered soap, solid soap or oil soap, the surfactant-containing cleansing formulation is a cleansing foam, cleansing water, cleansing towel or cleansing pack, the surfactant-free cleansing formulation is a cleansing cream , Cleansing lotion, cleansing water or cleansing gel, but is not limited thereto.

According to a preferred embodiment of the present invention, when the cosmetic method of the present invention is applied directly to the skin of a person, excellent anti-inflammatory, antibacterial and atopic improvement effect is excellent.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

Example  One

500 g of completely dried Sancho, Baekduong, moss, octagonal fennel, echinacea and propolis were collected in a ratio of 1: 1: 1: 1: 1: 1, put in 2kg of water, extracted at 95 ~ 100 ℃ for 2 hours, and filtered. It dried and 61g of hydrothermal extracts were obtained, respectively.

Example 2

500 g of completely dried Sancho, Baekduong, moss, octagonal fennel, echinacea and propolis were collected in a ratio of 1: 1: 1: 1: 1: 1, put in 2kg of ethanol, extracted for 2 hours at 95 ~ 100 ℃, and filtered. Drying yielded 59 g of ethanol extract, respectively.

Comparative Example 1

A total of 500g of completely dried Sancho, Baekduong, moss and octagonal fennel were collected in a ratio of 1: 1: 1: 1, put in 2kg of water, extracted at 95 ~ 100 ℃ for 2 hours, filtered and dried to obtain 43g of hydrothermal extract. .

Comparative Example 2

A total of 500g of completely dried Sancho, Baekduong, moss, octagonal fennel and echinacea were collected in a ratio of 1: 1: 1: 1: 1, put in 2kg of water, extracted for 2 hours at 95 ~ 100 ℃, filtered and dried and then dried for 49g each. Hot water extract was obtained.

Comparative Example 3

Collect 500 g of completely dried Sancho, Baekduong, moss, octagonal fennel and propolis in a ratio of 1: 1: 1: 1: 1, put it in 2kg of water, extract it for 2 hours at 95 ~ 100 ℃, filter it and dry it, 53g each. Hydrothermal extract was obtained.

Comparative Example 4

A total of 500 g of completely dried Sancho, Baekduong, moss and octagonal fennel were collected in a ratio of 1: 1: 1: 1, put in 2kg of ethanol, extracted for 2 hours at 95-100 ° C, filtered and dried to obtain 39g of ethanol extract, respectively. .

Comparative Example 5

A total of 500g of completely dried Sancho, Baekduong, moss, octagonal fennel and echinacea were collected in a ratio of 1: 1: 1: 1: 1, put in 2kg of ethanol, extracted for 2 hours at 95 ~ 100 ℃, filtered and dried to obtain 46g each. Ethanol extract was obtained.

Comparative Example 6

500 g of completely dried Sancho, Baekduong, moss, octagonal fennel and propolis were collected in a ratio of 1: 1: 1: 1: 1, put in 2kg of ethanol, extracted for 2 hours at 95 ~ 100 ℃, filtered and dried and 50g each. An ethanol extract of was obtained.

Experimental Example 1. Measurement of viable cell count

The antimicrobial effect of each mixed extract obtained in Example 1-2 and Comparative Example 1-6 was measured using a viable cell count method. First, each strain was cultured in Trypticase soy broth for 1 day and the culture conditions are described in Table 1. 20 ml Trypticase soy broth was put into a 100 ml flask and sterilized, and 100 µl was added from the seed medium. Mixed extracts of Example 1-2 and Comparative Examples 1-6 were added to 0%, 0.3% and 0.5%, respectively, and incubated at 25 ° C for 2 days. At this time, the number of viable cells of each bacteria in the activated culture was measured, and is shown in Table 2.

Viable cell count result sample density(%) Cell count (cfu / ml) E. coli P. aeruginosa S. aureus B. subtilis Control group 0.0% 230000000 78000000 550000000 420000000 Example 1 0.3% 0 1300 0 0 0.5% 0 0 0 0 Example 2 0.3% 0 0 0 0 0.5% 0 0 0 0 Comparative Example 1 0.3% 6800000 6200000 6100000 5900000 0.5% 74000 150000 180000 490000 Comparative Example 2 0.3% 920000 5600000 810000 840000 0.5% 3800 210000 5400 55000 Comparative Example 3 0.3% 80000 65000 48000 57000 0.5% 0 1600 4500 0 Comparative Example 4 0.3% 3900000 5900000 1800000 150000 0.5% 160000 380000 54000 29000 Comparative Example 5 0.3% 950000 1500000 81000 31000 0.5% 46000 28000 2800 9200 Comparative Example 6 0.3% 0 59000 9100 0 0.5% 0 9100 0 0

As shown in Table 2 and as shown in Table 2, in the result of measuring the viable cell count after treatment of the mixed extracts of Examples 1-2 and Comparative Examples 1-6 for the four strains disclosed, Sancho, Baekduong, 0.5% of mixed extracts extracted from six natural products of moss, octagonal fennel, echinacea and propolis showed excellent antimicrobial effect.

Experimental Example 2. Antimicrobial activity effect on microorganisms (minimum inhibitory concentration measurement method)

The antimicrobial activity of the mixed extracts of Examples 1-2 and Comparative Examples 1-6 against microorganisms was determined by a 96-well microdilution minimal inhibitory concentration assay. First, bacteria and fungi were incubated at 37 ° C. and 30 ° C. in TSB (trypticase soy broth) and SAB (Sabroaud, Saboraud), respectively, and then inoculated in fresh medium for 2 hours. The strains were logarithmic. After diluting them to 10 ⁵ strains per 1 ml, inoculating 190 μl into 96-well plates, and sequentially diluting the mixed extracts of Example 1-2 and Comparative Example 1-6 to 10 μl in each well. It was. The 96-well plate was incubated for 12 hours, and the absorbance of each well was measured to determine the minimum concentration at which the strain did not grow at all, as shown in Table 3 below.

 Antimicrobial Activity by the Minimum Inhibitory Concentration Method Strain MIC (ppm) Example 1 Example 2 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Comparative Example 5 Comparative Example 6 Gram Negative Bacteria Esherichia coli 160 130 460 300 260 450 310 230 Pseudomonas aeruginosa 150 190 460 340 310 430 340 280 Salm . Typhimurium 160 230 510 360 340 500 350 290 Salm . Pullorm 180 130 530 310 290 490 370 260 Aerobacter aerogenes 190 180 490 420 380 530 400 340 Proteus vulgaris 210 160 570 380 320 540 410 340 Vibrio cholerae 180 210 530 400 360 500 460 370 Cloaca cloacae 210 250 540 430 380 480 380 290 Klebsiella edwardsii 290 130 590 460 370 560 430 360 Legionella pneumoniae 270 200 610 480 380 580 400 340 Moraxella duplex 230 110 570 420 360 550 420 310 Shigella flexneri 240 160 580 390 340 540 410 320 Gram
Positive Bacteria Bacillus subtilis 180 150 600 400 290 590 430 300 Bacillus cereus 170 160 560 370 280 560 390 290 Clostridium botulinum 200 110 540 460 300 570 410 340 Lactobacillus arabinosus 240 220 600 430 370 560 480 310 Staphylococcus aureas 190 170 480 390 260 500 370 250 Listeria monocytogenes 210 190 430 370 290 420 360 310 Corynebacterium acnes 180 160 460 310 250 430 380 260 Diplococcus pneumoniae 290 270 540 420 350 500 400 310 Mycobacterium smegmatis 210 180 460 380 260 510 310 250 Sarcina lutea 260 230 560 460 360 530 420 330 Streptococcus agalactiae 160 150 640 410 240 580 390 260 Clostridium tetani 190 180 490 360 290 480 420 300 Fungus Aspergillus niger 290 260 690 590 360 730 560 350 Aspergillus fumigatus 310 360 680 680 400 700 580 420 Penicilium citrinum 360 370 720 630 530 700 600 460 Fusarium monliforme 200 220 700 650 380 690 590 380 Aureobasidium pullulans 200 240 690 640 400 730 610 290 Chaetomuim globosum 310 360 720 600 430 700 590 460

As shown in Table 3, as a result of measuring the minimum inhibitory concentrations of the mixed extracts of Example 1-2 and Comparative Examples 1-6 for 30 strains disclosed, Sancho, Baekduong, Moss, Octagonal Fennel, Echinacea and Pro In the case of extracts obtained by mixing and extracting all six natural products of polis, the lowest inhibitory concentration value was the lowest (for example, the minimum inhibitory concentration in bacteria was 100-300 ㎍ / ml and the minimum inhibitory concentration in mold was 200-400 ㎍ / ml Etc). This indicates that 6 natural mixed extracts of Sancho, Baekduong, Moss, Octagonal Fennel, Echinacea and Propolis have about 2-4 times better antimicrobial effect than less than 6 other mixed extracts.

Experimental Example  3. Antimicrobial activity effect against microorganisms (paper Disc method )

The antimicrobial activity of the mixed extracts of Examples 1-2 and Comparative Examples 1-6 against microorganisms was measured by the paper disk method as described below according to Zaika's method. First, after incubating the bacteria in a liquid medium, the number of bacteria was measured and plated in the same amount in a plate count Agar (PCA) solid medium and inoculated and incubated for 48 hours at 37 ℃ anaerobic. After making a hole having a diameter of about 1 cm in the plate inoculated with each bacterium, 100 μl of the diluted solution diluted 0.5% of distilled water with the mixed extract of Comparative Example 1-6 and Example 1-2 was dropped therein. 24 hours (for bacteria) or 48 hours (for molds) were further incubated. The diameter of the resulting clear zone was measured and shown in Table 4.

Antimicrobial Activity by Paper Disk Method Strain Clear zone (mm) Example 1 Example 2 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Comparative Example 5 Comparative Example 6 Gram Negative Bacteria Escherichia coli 29 29 10 14 14 11 14 13 Pseudomonas aeruginosa 20 20 10 13 12 12 15 14 Salm. typhimurium 18 18 11 14 16 13 15  16 Salm. pullorum 26 26 9 14 15 10 14 13 Aerobacter aerogenes 20 19 8 11 14 10 13 14 Proteus vulgaris 19 19 9 13 13 11 15 16 Vibrio cholerae 18 19 10 14 15 11 13 14 Cloaca cloacae 20 19 8 12 11 12 14 15 Klebsiella edwardsii 19 18 10 14 16 11 15 13 Legionella pneumoniae 16 17 11 13 15 10 14 14 Moraxella duplex 20 20 10 15 14 9 13 11 Shigella flexneri 17 18 10 14 16 10 14 13 Gram
Positive Bacteria Bacillus subtilis 16 19 9 10 11 10 14 15 Bacillus cereus 20 18 9 12 14 11 13 15 Clostridium botulinum 15 16 10 13 11 9 12 11 Lactobacillus arabinosus 16 18 10 11 14 10 13 14 Staphylococcus aureas 27 28 11 14 12 10 14 13 Listeria monocytogenes 25 26 9 12 13 8 11 14 Corynebacterium acnes 16 18 9 13 10 9 13 13 Diplococcus pneumoniae 18 19 10 14 11 11 14 14 Mycobacterium smegmatis 16 18 11 13 10 8 11 13 Sarcina lutea 17 19 10 14 14 8 10 12 Streptococcus agalactiae 16 16 9 12 10 9 10 11 Clostridium tetani 18 19 8 13 11 7 11 11 Fungus Aspergillus niger 16 18 10 14 12 8 12 13 Aspergillus fumigatus 14 16 8 11 12 9 11 12 Penicilium citrinum 13 14 8 10 11 7 11 10 Fusarium monliforme 14 16 7 10 10 8 10 11 Aureobasidium pullulans 11 12 7 10 10 7 9 10 Chaetomuim globosum 12 12 6 11 10 8 9 10

In the antimicrobial effect of the 30 strains disclosed, as shown in Table 4, as a result of measuring the diameter of the transparent ring for the microorganism of the mixed extract of Example 1-2 and Comparative Example 1-6, Sancho, Baekduong, moss In the case of mixed extracts of six natural products (Examples 1 and 2) of octagonal fennel, echinacea and propolis, the antimicrobial effect is much better than expected in the case of less than 6 mixed extracts of the natural products. Indicated. In particular, Salmonella Pullorum, Escherichia coli, Staphylococcus aureus and Listeria monocytogenes form a highly transparent antimicrobial effect, such as 25 mm or more. Indicated.

Experimental Example 4. Anti-inflammatory effect (ear edema experiment)

The anti-inflammatory effect was measured as follows using the mixed extract obtained in Comparative Example 1-6 and Example 1-2. With the left ear of the mouse as the control site and the right ear as the test site, the ears were washed thoroughly with ethanol before each mixed extract sample was applied. 20 μl of the extract sample was continuously applied to both ears once a day for 4 days, and 1 mg after the last application, ethanol was applied to the left ear and arachidonic acid (arachidonic acid) was applied to the right ear 2 mg / ear. After 1 hour, the ear edema was measured by repeating the measurement of both ears three times with a micrometer. The anti-inflammatory effect was determined as the degree of edema inhibition based on the arachidonic acid treatment group, which is a edema causing substance.

In Equation 1 below, A is the difference in the average thickness of the control ear (the thickness of the arachidonic acid-treated ears-the thickness of the ethanol-treated ears), B is the difference of the thickness of the extract coating group ears obtained in Comparative Examples 1-6 and Examples 1-2 (Ear thickness of arachidonic acid treatment group-thickness of ear of ethanol treatment group).

Anti-inflammatory Effects of Extracts sample Final concentration (% by weight) contained in the reaction solution Glume (㎛) % Inhibition Ethanol treatment
(Normal ear) Arachidonic acid treatment
Edema Control (purified water) - 301 573 - Example 1 One 297 461 40 Example 2 One 288 473 32 Comparative Example 1 One 286 513 17 Comparative Example 2 One 290 521 15 Comparative Example 3 One 300 506 24 Comparative Example 4 One 299 503 25 Comparative Example 5 One 280 506 17 Comparative Example 6 One 302 498 28

As shown in Table 5, as a result of measuring the anti-inflammatory effect of the mixed extracts of Example 1-2 and Comparative Examples 1-6, six kinds of the acidic, baekduong, moss, octagonal fennel, echinacea and propolis of the present invention In the case of extracts obtained by mixing and extracting natural products (Examples 1 and 2), it showed an excellent anti-inflammatory effect compared to the mixed extracts of Comparative Examples 1-6.

Experimental Example  5. Immune inhibitory effect

The immunosuppressive ability of the mixed extract obtained in Example 1-2 and Comparative Example 1-6 was measured as follows. Immune response inhibitory activity was confirmed by a mixed lymphocytes response assay. Briefly, splenocytes were isolated from spleens of BALB / c mice and C57BL / 6 mice. Separated cells were resuspended in RPMI-1640 medium, and splenocytes isolated from one mouse were treated with mitomycin C to stop cell division and washed several times with RPMI-1640 medium. Each adjustment was made. Each cell was mixed and added to a 96-well plate with a cell number of 1 × 10 ⁶ cells / well, followed by adding a sample and conA (concanavalin A), and incubating at 37 ° C. and 5% CO _{2 for} 4 days. XTT reagent was added 4 hours before the end of the incubation of splenocytes and the absorbance was measured at 490 nm after 4 hours of incubation. The immunosuppressive ability was determined by the rate of inhibiting cell differentiation compared to the control. In this case, cyclosporin A was used as a positive control, and the results of measuring the immunosuppressive ability are shown in Table 6 below.

In Equation 2, ODmax is the proliferation of cells when ConA and medium are added, ODmin is the proliferation of cells when only medium is added without adding ConA, and ODsample is a case where ConA and a sample are added simultaneously. Is the proliferation of cells.

Immunosuppressive effect sample Immunosuppressive effect (%) 1 mu g / ml 5 µg / ml 10 µg / ml 50 µg / ml 100 µg / ml Cyclosporin A - - 40 62 94 Example 1 10 31 54 87 98 Example 2 13 27 47 82 98 Comparative Example 1 5 19 40 67 74 Comparative Example 2 7 26 42 66 76 Comparative Example 3 6 25 43 69 74 Comparative Example 4 2 21 40 65 75 Comparative Example 5 2 23 43 67 74 Comparative Example 6 2 20 46 68 75

As shown in Table 6, the mixed extracts of Examples 1-2 and Comparative Examples 1-6 were generally higher in immunosuppressive activity than cyclosporin A, a substance known to have an immunosuppressive effect, and increased with increasing concentration. Also gradually increased. In particular, at the same concentration, the extract (Example 1 and Example 2) obtained by mixing and extracting all six species of Sancho, Baekduong, Moss, Octagonal Fennel, Echinacea and Propolis showed a much better effect than other mixed extracts. .

Preparation Example 1 Preparation of Hand Wash Cosmetic

According to the composition of Table 7, a hand wash cosmetics containing a mixed extract of Sancho, Baekduong, moss, octagonal fennel, echinacea and propolis was prepared. The mixed extract used in the formulation was used extracts (Examples 1 and 2) using purified water and ethanol as solvents, respectively, with the use of Sancho, Baekduong, Moss, Octagonal Fennel, Echinacea, and Propolis. The content unit is g.

sample Formulation Example 1 Formulation Example 2 Mixed extract of Example 1 0.5 - Mixed extract of Example 2 - 0.5 glycerin 5 5 Camidopropylbetaine 3 3 Decylglucoside 10 10 Potassium Coyl Glycinate 5 5 Disodium iodide 0.02 0.02 Polyquaternium-7 0.5 0.5 ethanol 2 2 Purified water to 100 to 100

Experimental Example 6. Bactericidal effect of cosmetic formulation

The bactericidal effect of the hand-washing formulation prepared in Preparation Example 1 was carried out with respect to an unspecified general person as follows. The measuring method was performed by pressing an unspecified hand of both hands in a state without washing at a constant pressure on the hand medium, using the composition, and then pressing the same hand again on the other hand medium in the same manner. At this time, for comparison, a general soap was used as a control instead of the cosmetic of the hand-washing formulation of the present invention. The bactericidal effect was measured in percentage by measuring the number of bacteria in the hands before and after the use of the cosmetic formulation and general soap of Preparation Example 1, the results are shown in Table 8. At this time, the bacteria used for the measurement are Staphylococcus aureus , Pseudomonas aeruginosa and Escherichia coli .

Bactericidal Effect of Cosmetic Formulations Material used Sterilization power before and after use (%) Staphylococcus aureus Pseudomonas aeruginosa Escherichia coli Control (normal soap) 98.01 98.00 98.13 Formulation Example 1 99.76 99.79 99.61 Formulation Example 2 99.89 99.86 99.69

As shown in Table 8, Formulation Example 1 and Formulation Example 2 of the cosmetic formulation of the present invention showed a bactericidal effect of 95% or more, as in the case of the general soap, whereby the acid, baekduong, moss, octagonal fennel, echinacea of the present invention And when the propolis mixed extract is applied to the cosmetics it can be seen that there is an excellent antibacterial effect.

Preparation Example 2 Preparation of Cream Formulation

According to the composition of Table 9, a cosmetic formulation of a cream formulation containing a mixed extract of Sancho, Baekduong, Moss, Octagonal Fennel, Echinacea and Propolis was prepared. As a mixed extract used in the formulation, extracts (Example 1 and Example 2) using purified water and ethanol as solvents, respectively, were prepared using Sancho, Baekduong, Moss, Octagonal Fennel, Echinacea, and Propolis as solvents. At this time, the content unit of each component of Table 9 is g.

sample Formulation Example 3 Formulation Example 4 Comparative Formulation Example 1 Comparative Formulation Example 2 Comparative Formulation Example 3 Comparative Formulation Example 4 Comparative Formulation Example 5 Comparative Formulation Example 6 Mixed extract of Example 1 One - - - - - - - Mixed extract of Example 2 - One - - - - - - Extract of Comparative Example 1 - - One - - - - - Extract of Comparative Example 2 - - - One - - - - Extract of Comparative Example 3 - - - - One - - - Extract of Comparative Example 4 - - - - - One - - Extract of Comparative Example 5 - - - - - - One - Extract of Comparative Example 6 - - - - - - - One glycerin 3 3 3 3 3 3 3 3 Acrylate / C10-30 Alkylacrylate Crosspolymer 0.06 0.06 0.06 0.06 0.06 0.06 0.06 0.06 Polyglyceryl-3methylglucose distearate 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Fiji-4 Olive Bay 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Cetearyl Alcohol 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 Squalene 4.5 4.5 4.5 4.5 4.5 4.5 4.5 4.5 Caprylic / capric triglyceride 4 4 4 4 4 4 4 4 Dimethicone 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Potassium Hydroxide Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Purified water to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100

Experimental Example 7. Atopic Treatment and Improvement of Cosmetic Formulation

Skin disease treatment effect on the cosmetics of the cream formulation prepared in Preparation Example 2 was measured as follows. 5 to 30 years old atopic patients suffering from atopic dermatitis for 1 year or more were applied to the skin once every 30 days after cleaning every evening to measure the degree of atopic dermatitis improvement. Fifteen subjects were assigned to the first group, and scored according to the four-point scale (4: very good, 3: moderately better, 2: slightly better, 1: no change, 0: worse). The average value is shown in Table 10 below. 30 days after starting the application of the present invention to atopic patients, the overall improvement of the symptoms of atopic dermatitis was divided into four stages to evaluate the patients or their parents in the form of a questionnaire. At this time, for comparison, a cosmetic composition containing no mixed extract was used as a control, and the results are shown in Table 10.

Atopic dermatitis improvement effect division Atopic dermatitis improvement effect Control group (mixed extract no addition) 1.4 Formulation Example 3 (Extract of Example 1) 3.7 Formulation Example 4 (Extract of Example 2) 3.8 Comparative Formulation Example 1 (Extract of Comparative Example 1) 2.8 Comparative Formulation Example 2 (Extract of Comparative Example 2) 2.5 Comparative Formulation Example 3 (Extract of Comparative Example 3) 2.5 Comparative Formulation Example 4 (Extract of Comparative Example 4) 2.5 Comparative Formulation Example 5 (Extract of Comparative Example 5) 2.5 Comparative Formulation Example 6 (Extract of Comparative Example 6) 2.4

As shown in Table 10, in the atopy treatment and improvement effect, the cosmetic composition containing the mixed extract of Example 1-2 and Comparative Examples 1-6 is atopic dermatitis treatment and improvement effect compared to the control group does not contain the mixed extract Was found to be excellent, and in particular, the cosmetics of Formulation Examples 3 to 4 containing the mixed extract of Example 1 and Example 2, which extracted all six natural products of Sancho, Baekduong, Moss, Octagonal Fennel, Echinacea and Propolis The composition was particularly effective in treating and improving atopy.

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that such a specific technology is only a preferred embodiment, and the scope of the present invention is not limited thereto. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (6)

A cosmetic composition for skin improvement comprising a mixed extract of Sancho, Baekduong, Lichen, Octagonal Fennel, Echinacea and Propolis as an active ingredient. According to claim 1, wherein the skin improvement is a cosmetic composition for skin improvement, characterized in that antibacterial, anti-inflammatory and atopy improvement. The cosmetic composition for improving skin of claim 1, wherein the mixed extract is contained in an amount of 0.1 to 10 wt% based on the total weight of the composition. According to claim 1, wherein the mixed extract is a cosmetic composition for skin improvement, characterized in that extracted with water or ethanol for 2 to 3 hours at 95-100 ℃. The cosmetic composition for improving skin of claim 1, wherein the cosmetic composition has a formulation selected from the group consisting of softening cream, nutrient cream, nourishing cream, massage cream, essence, pack, or emulsified foundation. delete