KR20100023478A - Composition comprising of plant extract having antimicrobial and antiseptic activity and the use thereof - Google Patents

Abstract

PURPOSE: A composition containing plant extract with antibacterial and preservative activities is provided to have broad antibacterial spectrum and apply to various products such as cosmetic, food, and medicinal and pharmaceutical products. CONSTITUTION: An antibacterial and preservative composition contains 1-50 weight% of extracts of Myristica fragrans, Eugenia aromatic, Eucalyptus alba, and Citrus paradise. The extracts are isolated from seed of Myristica fragrans, Eugenia aromatic, Eucalyptus alba, and Citrus paradise using water; alcohol such as ethanol, methanol, or butanol, and organic solvent such as ethylene, acetone, ether, hexane, chloroform, or ethyl acetate.

Description

COMPOSITION COMPRISING OF PLANT EXTRACT HAVING ANTIMICROBIAL AND ANTISEPTIC ACTIVITY AND THE USE THEREOF}

The present invention relates to a cosmetic composition exhibiting a broad antimicrobial spectrum against various strains, such as acne, dandruff, fungi and yeast, has an antioxidant activity and does not exhibit toxicity and can be applied to various cosmetic compositions.

Cosmetics consist of an aqueous phase containing water and water soluble ingredients and an oily fraction containing oils, waxes and fat soluble ingredients. The moisture and various organic substances in these products act as good nutrients for bacteria and fungi, so microorganisms easily reproduce. Cosmetics contaminated with microorganisms have a risk of skin troubles and microbial infections such as skin rashes and inflammation as well as deterioration due to alteration of ingredients. Therefore, most cosmetics contain a preservative for the purpose of inhibiting or killing the growth of microorganisms. The role of these preservatives in cosmetic compositions is even more important, especially because of their long service life from months to years.

Preservatives currently used in cosmetics include methyl-, butyl-, propyl-parabens (ester of para-hydroxybenzoic acid), phenoxyethanol, imidazolidinyl urea, diazolidinyl urea, sorbic acid, or the like. It has been used mainly.

These artificial preservatives are used in most cosmetics and household goods because of their ease of use and excellent antiseptic effect. However, as various problems and side effects of artificial preservatives have been recently discovered, a danger arises when used excessively or excessively.

Problems with artificial preservatives include (1) skin irritation or allergy (the American Academy of Dermatology designates preservatives as a potential source of skin allergy), (2) the likelihood of cancer (parabens in breast cancer tissues, Journal of Applied Toxicology , 2004, 24: 1, 1-4), ③ Weakened male fertility, the role of female hormones (many preservative related studies such as parabens), ④ In addition, there are concerns about the possibility of central nerve palsy, genetic mutations, etc. WHO also warns of the risk of side effects .

Due to the potential risks of such artificial preservatives, researches have recently been actively conducted to introduce different kinds of preservatives or to find substances having antibacterial or antiseptic effects among substances existing in nature.

For example, organic acids such as succinic acid, malic acid, tartaric acid, and benzoic acid contained in natural products inhibit the growth of microorganisms, and flavonoids and catechin compounds have an antimicrobial effect, and the carbon number of 12 -18 fatty acids have been reported to be effective antimicrobials. In addition, the antimicrobial effect on proteinaceous substances, such as lysozyme in eggs, lactoferrin in milk and lactobacillus, and lactic acid bacteria are known.

In Korea, the antibacterial effect of garlic and onion, which is a widely used spice, has been studied, and the antibacterial activity of various herbal or plant ingredients in Korea has been studied. It was reported to have strong antimicrobial activity against both gram positive and negative bacteria. In addition, it has been reported that the extract of S. vulgaris inhibits the proliferation of Listeria monocytogenes , and that the herbal medicines Schisandra chinensis, turmeric, vinegar, red ginseng, licorice and the like have antimicrobial activity.

Recently, with the trend of naturalism in the cosmetics industry, problems of existing preservatives have emerged, and raw material companies and cosmetic product companies are actively developing products using unprotected or natural preservative systems.

For example, a raw material product using wormwood, silver (Ag) nano ingredients, pine needles, and antibacterial peptides derived from lactic acid bacteria, in addition to products using herbal preparations such as complex preparations using seven plant ingredients such as green tea as extract ingredients and herbal ingredients such as Scutellaria baicalensis Some of these lamps are being used or are under continuous development, and each cosmetic product company is trying to use a natural preservative system.

However, the development of effective antiseptic / antibacterial agents requires more extensive search and research on the natural products available at home and abroad. In particular, there is a need for the development of natural antimicrobial / preservatives having the broadest antimicrobial / preservative spectrum, formulation stability, and the like, while reducing side effects, which are the biggest disadvantages of antimicrobial / preservatives.

The inventors of the present invention screened those that have excellent antimicrobial and antiseptic activity in plant species native to the tropics, and carried out antimicrobial and antiseptic screening on these plant species, among which the antimicrobial and antiseptic activity were excellent and antibacterial by synergistic effect in combination. And a broad spectrum spectrum of antiseptics, and confirmed plant extracts having antibacterial and antiseptic activity safe to humans. In addition, as a result of further research to study the extraction method to further improve the antibacterial and antiseptic activity of the plant extracts, through the fermentation process by lactic acid bacteria in the extraction process, the antimicrobial and antiseptic activity was found to be further improved and more effective natural Antimicrobial and preservatives have been developed.

An object of the present invention is that the composition exhibits a broad antimicrobial spectrum against various strains, such as acne, dandruff, fungi and yeast, has an antioxidant activity and does not exhibit toxicity and can be applied to various cosmetics, food, medicine, pesticides or household products It is to provide antimicrobial and antiseptic compositions and uses thereof.

In order to achieve the above object, the present invention provides nutmeg ( Myristica fragrans ), cloves ( Eugenia aromatica ), Eucalyptus alba ), and grapefruit ( Citrus) paradisi ) provides an antimicrobial and antiseptic composition comprising the extract.

The present invention also provides cosmetics, food, pharmaceuticals, pesticides or household goods containing the antimicrobial and antiseptic composition.

The present invention also provides a cosmetic composition having antimicrobial and antiseptic activity, including nutmeg ( Myristica fragrans ), cloves ( Eugenia aromatica ), Eucalyptus alba , and grapefruit ( Citrus paradisi ) extract.

The composition comprising the plant extract according to the present invention exhibits a broad antimicrobial spectrum against a variety of bacteria, and is excellent in antioxidant activity and does not generate any toxicity or irritation in various safety tests, and thus is safe for human use. Such a composition is applicable to cosmetic compositions such as cosmetics, in addition to food, pharmaceuticals, pesticides and household goods, etc. can be usefully used for sterilization, antibacterial and antiseptic purposes.

The composition according to the invention comprises a plant extract and exhibits antibacterial and antiseptic effect.

Specifically, the plant extracts are nutmeg ( Myristica fragrans ), cloves ( Eugenia aromatica ), Eucalyptus ( Eucalytus alba ), and grapefruit ( Citrus paradisi ) extract, and antimicrobial and antiseptic compositions comprising the same are provided herein. Preferably they are extracts of their leaves, seeds or stems, and more preferably extracts from nutmeg seeds, cloves seeds, eucalyptus leaves and grapefruit seeds.

Nutmeg is a dried fruit of the evergreen tree, Myristica fragrans , which belongs to the nutmeg family. It is native to the Moleka Islands of Indonesia and is called 'pala' in English and 'nutmeg' in English, meaning a walnut with a musk fragrance. Have It grows well in hot and humid climates. Seeds are nucleus, 4-6 cm long, look apricot when mature and contain seeds inside. The nutmeg contains myristin (lipid) essential oils such as myristicin, pinene, and camphene, and is used as a fragrance forgery or tonic, or as a spice in the West.

Olajide et al . Reported that nutmeg extracts had anti-inflammatory effects [Olajide et al , Biological effects of Myristica fragrans (nutmeg) extract, Phytotherapy Research , 13 (4): pp 344-345, 1999), and Dinesh Dhingra et al. n-hexane extracts have been reported to have antidepressant effects [Dinesh Dhingra et al , Antidepressant-Like Activity of n-Hexane Extract of Nutmeg (Myristica fragrans) Seeds in Mice, Journal of Medicinal Food , 9 (1): 84- 89, 2006].

Cloves are dried buds of clove, which is native to the Indonesia Molucca Islands, also called Eugenia caryophyllata , Syzygium aromatica , and is called "cengken" in the original language. The original language is called "cengken". Cloves are 4-7m in height, seeds are nucleus, black red, and seeds, flower stalks, and buds before flowering. In addition, since the clove is very fragrant, it is used as it is or as a powder, and it uses clove oil (Eugenol, Eugenol) extracted with water or steam, and is used for food, medicine or antiseptic effect, or as a painkiller in dentistry and dentistry. .

Sanchez-Perez J et al . Have reported that eugenol, cinnamon oyl and clove oil are effective in skin allergy [Sanchez-Perez J et al , Occupational allergic contact dermatitis from eugenol, oil of cinnamon and oil of cloves in a physiotherapist. Contact Derm , 41 (6): 346-347, 1999], Guynot ME et al . Suggest that cloves have antifungal activity [Guynot ME, Ramos AJ, Seto L, et al . Antifungal activity of volatile compounds generated by essential oils against fungi commonly causing deterioration of bakery products. J Appl Microbiol , 94 (5): 893-899, 2003].

Eucalyptus is called "Daun kayu putih" in Indonesian and is a kind of evergreen eucalyptus tree, which is strong in drought and cold. Eucalyptus is called white gum, and wood, gum and oil are used as folk medicines and burned ash is used as a mosquito scent.

Atta et al . Reported that eucalyptus extracts have anti-inflammatory activity [Atta et al , Anti-nociceptive and anti-inflammatory effects of some Jordanian medicinal plant extracts. J Ethnopharmacol , 60 (2): 117-24. Hideki et al , Effect of Eucalyptus Extract Chewing Gum on Periodontal Health: A Double-Masked, Randomized Trial, J of Periodontology, 79 (8). : 1378-1385, 2008].

Grapefruit is the fruit of the grapefruit tree belonging to the citrus, also called "jabong", "Jeruk Gripfruit" or "Jeruk bali". Grapefruits have a grape-like scent, run like grapes, round in tangerine shape, 10-15 cm in diameter, and their outer skin is smooth and the surface is smooth and yellow. The pulp is pale yellow, rich in juice, sour, sweet and bitter, and has recently developed a pink pulp.

Laura Giamperi et al . Have reported that grapefruit extract has antioxidant effect [Laura Giamperi et al , Antioxidant activity of Citrus paradisi seeds glyceric extract, Fitoterapia , 75 (2): 221-224, 2004], and Adeneye et al . Adeneye et al , Haematopoetic effect of methanol seed extract of Citrus paradisi Macfad (grape fruit) in Wistar rats, Biomedical Research , 19 (1): 23-26, 2008.

These plant extracts are known to be edible and medicinal as well as antiseptic in some but are difficult to use as a broad range of potent antiseptic or antimicrobial agents commercially available as single plants or extracts. Accordingly, the present inventors have completed the present invention by confirming the antibacterial and antiseptic activity of each of the extracts of these plants and using them in combination at an appropriate ratio.

The plant extract included in the composition of the present invention may extract each plant using water, an organic solvent, or a mixed solvent thereof. Specifically, water; Alcohols such as methanol, ethanol, isopropanol or butanol; Extraction is possible using each of organic solvents such as ethylene, acetone, ether, hexane, chloroform, or ethyl acetate or a mixed solvent thereof.

The extraction follows a conventional extraction process and can be extracted at room temperature or by heating. Preferably it is carried out in a solvent 2 to 10 times the weight of the raw material, preferably in a 50 to 80% ethanol solution, preferably at a temperature of 50 ℃ to 70 ℃, preferably for 2 to 4 hours.

In addition, during extraction, all of these components may be mixed and extracted at the same time, or individual components may be extracted and mixed under optimum conditions. When heating at a high temperature, the time is reduced, but the active ingredient is destroyed and can be lost. When heating at a temperature lower than the temperature, it is important to control the temperature and time properly because the active ingredient of the mixture is not completely extracted.

The extract obtained through the extraction process is centrifuged or filtered to remove impurities by a conventional method, and the filtrate is used by distilling an organic solvent and then drying it or dissolving it in a 50% solution of 1,3-butylene glycol.

On the other hand, the present inventors confirmed that the antimicrobial and antiseptic ability are further improved through the fermentation process by lactic acid bacteria during the production of antimicrobial and antiseptic activity in these plant species.

That is, the components having antimicrobial activity in plants are mainly composed of phenolic compounds such as essential oils, alkaloids and flavonoids, and the function of the components is not only the type and amount of these components, but also the extracted state, that is, the non-glycoside / glycoside (Glycoside / It also varies greatly depending on the aglycone state. In the extraction process of the nutmeg, cloves, eucalyptus and grapefruit extract, by adding a fermentation method rather than a simple extraction, it enhances the extractability of the active ingredient in solid plant tissues and changes the state of the extract component present as a glycoside to a non-saccharide. It further enhances antibacterial and antiseptic properties.

The fermentation is a strain of Bifidobacterium sp, including Bifidobacterium longum , known as a beneficial strain in human or animal intestines, and Lactobacillus bulgaricus , including Lactobacillus bulgaricus . ( Lactobacillus sp. ) Strain is effective.

The plant extract thus obtained is used in a ratio of 1: 2: 2: 1 by weight ratio (solid content) of nutmeg: clove: eucalyptus: grapefruit, and this content can be adjusted as necessary.

In particular, the compositions according to the invention exhibit a wide range of antimicrobial activities against various microorganisms, in particular bacteria, fungi and yeast. Specifically, the composition is a bacterium such as Escherichia coli (KCTC 2571), Staphylococcus aureus KCTC 1916, Pseudomonas aeruginosa KCTC 2513, Candida albicans KCTC 7965) and antifungal activity against fungi such as Aspergillus niger KCTC 6911.

In addition, the composition not only has an antioxidant effect, but also various tests using rats and rabbits confirmed that the skin is safe and does not exhibit acute oral toxicity.

On the other hand, the present invention provides the use of a composition comprising nutmeg ( Myristica fragrans ), cloves ( Eugenia aromatica ), Eucalyptus alba , and grapefruit ( Citrus paradisi ) extract.

The composition may be applied to cosmetics, food, pharmaceuticals, pesticides or household goods, preferably cosmetics as a cosmetic composition.

As described above, the antimicrobial / preservative composition according to the present invention having a broad antimicrobial spectrum can be widely used to achieve the purpose of antibacterial, sterilization, disinfection, antiseptic, etc. in food as well as food, household goods, pesticides, medicines and the like.

Specifically, in cosmetics and household goods, it is used in products directly related to microorganisms, such as dandruff suppression, athlete's foot prevention, armpit harvest control, and anti-acne, and is used for preservatives, antibacterial or sterilization for small tablets or dishwashing detergents. Can be done. In addition, food can be used for food preservation and antibacterial purposes, agriculture for antibacterial, sterilization, and disinfection purposes, and for medicine, such as antibiotics and antifouling agents, but is not limited to these purposes.

In particular, the composition according to the present invention can be used as a cosmetic composition.

According to the experimental example of the present invention, the composition shows an effective antimicrobial activity against the acne bacteria Propionibacterium acne ( propionibacterium acne) , it can be used to improve acne skin.

In addition, compositions of the invention may show an effective antimicrobial activity even in the error correcting capability of Ross bideumgyun Forum misfire (Pityrosporum ovale), by applying to the hair and scalp, effectively inhibit bideumgyun.

The composition according to the present invention also showed an excellent antioxidant effect compared to ascorbic acid, which is a conventionally known antioxidant. In addition, the composition of the rat oral test, rabbit eye mucosal irritation test, skin irritation test and phototoxicity test using a guinea pig, it was confirmed that there is no toxicity and no irritation to the eyes, skin and the like.

The cosmetic composition according to the present invention can be applied to various cosmetics, and is not particularly limited in the present invention.

For example, it may be a cosmetic having a formulation of a flexible lotion, a nourishing lotion, a massage cream, a nourishing cream, a pack or a gel, an external product having a formulation of a body lotion, an ointment, a gel, a cream, a patch or a spray, It can be used as an artificial antibacterial agent for shampoo, body shampoo, rinse, cleansing, face wash, soap, treatment, and essence.

Example 1 Preparation of a Composition Comprising a Mixed Extract

Nutmeg seeds, cloves seeds, eucalyptus leaves and grapefruit seeds 1kg each was shredded with a shredder, and 10kg of 70% ethanol was extracted by heating at 60 ° C. for 3 hours using a reflux apparatus. The obtained extract was centrifuged to remove the precipitate, distilled to remove the solvent, and mixed with 10 kg of 50% 1,3-butylene glycol, respectively, to prepare an extract solution.

Each extract solution was made to be nutmeg seeds (3% by weight), cloves seeds (6% by weight), eucalyptus leaves (6% by weight), and grapefruit seeds (3% by weight). Subsequently, these extract solutions were mixed to prepare a composition including the mixed extract.

Example 2: Preparation of a Composition Comprising a Mixed Fermented Extract

1 kg each of nutmeg seeds, cloves seeds, eucalyptus leaves and grapefruit seeds are shredded with a shredder, and added with the same amount of water and 10% weight of sugar, followed by Bifidobacterium longum , a lactic acid bacterium. Inoculated 1% by weight. Inoculated samples were sealed and fermented at 15 ° C. or lower for 30 days. After fermentation, 70% ethanol was added to 10 times the weight of the sample, and the mixture was heated and extracted at 60 ° C. for 3 hours using a device equipped with a reflux device.

The obtained extract was centrifuged to remove the precipitate, distilled to remove the solvent, and mixed with 10 kg of 50% 1,3-butylene glycol, respectively, to prepare an extract solution.

Each extract solution was made to be nutmeg seeds (3% by weight), cloves seeds (6% by weight), eucalyptus leaves (6% by weight), and grapefruit seeds (3% by weight). Subsequently, these extract solutions were mixed to prepare a composition including the mixed fermentation extract.

Experimental Example 1: Evaluation of antimicrobial efficacy by the diffusion method

Escherichia coli (KCTC 2571), Staphylococcus aureus KCTC 1916, Pseudomonas aeruginosa as a bacterium to investigate the antimicrobial activity spectrum of the composition comprising the mixed extract obtained in Example 1 and the individual extracts Pseudomonas aeruginosa KCTC 2513 was added to Nutrient agar (Difco), Candida albicans KCTC 7965 as yeast, and Aspergillus niger KCTC 6911 as fungus , respectively. (Sabouraud dextrose agar, Difco) was used in culture.

As the first antimicrobial activity, the antimicrobial activity was measured by the agar diffusion method. A number of disk filter papers were coated with a known concentration of extract solution, placed on a solid agar medium coated with the test strain, and cultured, and then the size of the growth inhibition halo around the disk was measured. In this case, the larger the zona pellucida, the stronger the antimicrobial activity, and the results obtained are shown in Table 1 below. As a control, phenonip (Clariant) widely used as a cosmetic preservative was used.

Antimicrobial Activity of Plant Extracts by Diffusion Method sample E. coli S. aureus P. aeruginosa C. albicans A. niger Nutmeg Extract ++++ +++ +++ +++ + Clove Extract ++ ++ ++++ ++++ +++ Eucalyptus extract + +++ +++ ++++ + Grapefruit extract +++ ++++ +++ +++ +++ Composition of Example 1 ++++ ++++ ++++ ++++ ++++ Control: Phenonip (0.2%) ++++ ++++ ++++ ++++ ++++ Size of the transparencies: +; 8-12 mm, ++; 13-16 mm, +++; 17-20 mm, ++++; 21-24 mm, +++++; 24 mm

As shown in Table 1, the nutmeg and grapefruit extract showed excellent antibacterial activity against Escherichia coli and Staphylococcus aureus, cloves and eucalyptus against Pseudomonas aeruginosa and Candida albican. In addition, clove and grapefruit extract showed excellent antifungal activity against the fungus Aspergillus niger.

Nutmeg, cloves, eucalyptus and grapefruit extracts presented in the present invention have a slight difference in antimicrobial and antifungal activity against each strain, but the extract of Example 1, which is a mixed extract thereof, showed a strong activity against all the strains presented. The synergistic effect was also observed for fungi.

Experimental Example  2: minimum inhibitory concentration by liquid culture method ( MIC ) decision

In order to measure the minimum inhibitory concentration (MIC) of the composition according to the invention was carried out as follows.

After incubating the strains shown in Table 2 in the respective culture medium (culture medium minus agar) and conditions, the samples were prepared by serial dilution on the medium, and the incubated culture solution was inoculated to 10 ⁷ cell / ml. . After culturing in each culture condition to check the growth of microorganisms, the minimum concentration (in terms of solid content) in which the bacteria do not grow was determined by MIC.

Minimum Inhibitory Concentration of Plant Extracts by Liquid Culture Method (MIC, Unit%) sample E. coli S. aureus P. aeruginosa C. albicans A. niger Nutmeg Extract 0.03 0.06 0.06 0.24 - Clove Extract 0.24 0.24 0.03 0.015 0.06 Eucalyptus extract 0.96 0.48 0.015 0.03 - Grapefruit extract 0.06 0.06 0.48 0.24 0.12 Mixed Extract of Example 1 0.06 0.12 0.06 0.12 0.12 Control: Phenonip (0.2%) 0.10 0.20 0.10 0.20 0.20 -: No antiseptic effect

As shown in Table 2, the nutmeg extract showed an excellent antimicrobial activity evenly in E. coli and Staphylococcus aureus and Pseudomonas aeruginosa. In addition, grapefruit extract showed relatively good antibacterial properties in Escherichia coli, Staphylococcus aureus, Clove and Eucalyptus extract in Pseudomonas aeruginosa and Candida albican.

Clove and grapefruit extracts were also effective against the fungus Aspergillus niger.

Similar to the results in the solid medium of Experimental Example 1, the composition containing the mixed extract of Example 1 showed a strong antimicrobial activity against all the strains, and also showed a synergistic effect on fungi.

Experimental Example  3: Antimicrobial Activity of Mixed Fermented Extracts

The antimicrobial activity of the fermented extracts prepared in Examples 1 and 2 was measured by the same diffusion method as in Experimental Example 1, and the results obtained are shown in FIG. 1.

Figure 1 is a photograph showing the antimicrobial activity of the composition prepared in Examples 1, 2 and the control. Referring to FIG. 1, when the compositions (D, E, F) of Examples 1 and 2 were used, the antimicrobial activity was excellent, and compared to the commercial control group (C), it can be seen that the same or more effects were obtained.

In addition, it can be seen that the composition of Example 2 obtained by performing the fermentation process shows an excellent antimicrobial activity in case of extract. These results are believed to be due to the conversion of antimicrobial components such as phenol components of plants into active molecules or low-molecular structures that are easy to penetrate into microbial cells.

Experimental Example 4: Determination of Minimum Inhibitory Concentration (MIC) by Liquid Culture

The minimum inhibitory concentration (MIC) for each strain was measured by the liquid culture method of the composition obtained through fermentation in Example 2, and the results are shown in Table 3 below.

Minimum Inhibitory Concentration of Plant Extracts by Liquid Culture Method (MIC, Unit%) sample E. coli S. aureus P. aeruginosa C. albicans A. niger Composition of Example 1 0.06 0.12 0.06 0.12 0.12 Composition of Example 2 0.015 0.6 0.06 0.06 0.12 Control group: Phenonip 0.10 0.20 0.10 0.20 0.20

Referring to Table 3, the fermented extract of Example 2 showed better antibacterial activity against Escherichia coli, Staphylococcus aureus, and Candida albican.

Experimental Example 5: Antimicrobial Activity against Acne Bacteria

Table 4 shows the results obtained by measuring the antimicrobial activity against propionibacterium acne ( Propionibacterium acne KCTC5012), the causative agent of acne among skin-derived bacteria.

P. acnes was used as a medium in which 0.1% Tween 80 was added to Reinforced Clostridial Medium (Difco, USA) and Brain Heart Infusion Medium (Difco, USA), and sealed using an anaerobic jar (with BBL Gaspak). Incubated for 3 days in a C0 ₂ incubator.

Minimum Inhibitory Concentration of Plant Extracts against Acne Bacteria (MIC, Unit%)  Group Acne Bacteria: Propionibacterium acne Composition of Example 1 0.12 Composition of Example 2 0.06

Referring to Table 4, it can be seen that the composition according to the present invention exhibits high antibacterial activity against acne bacteria. In addition, in the case of the composition of Example 2, including the fermentation extract exhibited superior activity than the composition of Example 1, inhibiting the growth of bacteria even at 1/2 concentration.

Experimental Example 6: Antibacterial activity against dandruff

In order to determine the antimicrobial activity against dandruff, Pyrosporum ovale ATCC 12087 was used to measure bacterial growth inhibition, and the results obtained are shown in Table 5 below.

P. ovale was cultured using Pytyrosporum medium (Malt Extract 6%, Ox-bile 2%, Tween 40 1%, Glycerol mono-oleae 0.25%).

Minimum Inhibitory Concentration of Plant Extracts against Dandruff (MIC, Unit%) sample Dandruff: Pityrosporum ovale Composition of Example 1 0.24 Composition of Example 2 0.24

Referring to Table 5, the compositions of Examples 1 and 2 according to the present invention showed a high antibacterial activity against dandruff. The compositions of Examples 1 and 2 effectively inhibited bacterial growth at 0.24%.

Experimental Example 7: Antioxidant Effect

Antioxidant activity measurement by electron donating ability (EDA) is a method of directly evaluating the reactivity of antioxidants against radicals in the model system of LOO. to be. Antioxidants donate electrons or hydrogen to free radicals, and DPPH forms molecules irreversibly stable by electrons or hydrogen received from antioxidants, and thus antioxidant activity can be measured from the electron donating ability. Electron donation was performed by adding 4x10 ^-4 M DPPH solution (2,2'-diphenyl-picrylhydrazyl) to 0.2 ml of each sample, shaking for 10 seconds with a vortex mixer and after 10 minutes UV / VIS spectrophotometer. , Jasco. Japan) was used to measure the absorbance at 525 nm. In this case, the electron donating effect was calculated as a percentage of the absorbance difference between the sample addition and the sample addition group, and the concentration (median scavenging concentration, SC50) indicating the electron donating ability of 50% was measured and the results obtained are shown in Table 6 below.

Antioxidant activity sample EDA (SC _50, mg / ml) Composition of Example 1 6.5 Composition of Example 2 5.2 Control group: Phenonip - Control Group 2: Ascorbic Acid 0.2 - : no effect

Referring to Table 6, the electron donating ability (SC ₅₀ ) of the compositions of Examples 1 and 2 was 6.5 mg / ml and 5.2 mg / ml, respectively, showed better values than the known antioxidant ascorbic acid.

When the composition according to the present invention is used as an antibacterial and preservative, it is possible to replace the existing antibacterial and preservatives only if there is no toxicity to the human body, and the use thereof can be broadened. In the following experimental example, the oral toxicity and the safety for the skin were tested using the composition prepared in Example 1.

Experimental Example 8: Single Oral Dose Toxicity Test in Rats

This test was performed to evaluate the toxicity of a single oral administration of the composition of Example 1 to rats, and to determine the approximate lethal dose of the test material in doses of 0 (control, water for injection) and 2000 mg / 10 mL / kg. Five mice were administered orally once, and the general symptoms, body weight changes, and gross findings at necropsy were observed for 15 days. The results obtained are as shown in Table 7 below.

Oral Dose Toxicity Test Results of the Composition of Example 1 gender Group / Dose (mg / kg) Number (mari) Days of Treatment Death rate (death / total) Dose amount (mg / kg) One 5 10 15 cock G1 0 5 0 0 0 0 0% (0/5) > 2000 G2 2000 5 0 0 0 0 0% (0/5) female G1 0 5 0 0 0 0 0% (0/5) > 2000 G2 2000 5 0 0 0 0 0% (0/5)

No mortality and body weight change due to administration were observed in both male and female controls and test groups. During the observation period, diarrhea and stool were observed in the cancer and water test group on day 1, but no specific general symptoms were observed from day 2, and gross abnormalities were observed in the cancer and water control group and test group. It wasn't.

As a result of oral administration of the composition of Example 1 to a rat once, no death by the test substance was observed, and the approximate lethal dose was 2000 mg / kg or more for each male and female.

Experimental Example 9: Eye irritation test using rabbit

The ocular mucosa stimulation test of the composition of Example 1 as a test substance was carried out using nine male 16-week-old male NZW rabbits. Test substance was administered as is. In the non-eye group, 0.1 mL of the test substance was administered into 6 right eye conjunctival sac, and eye lesions of the cornea, iris and conjunctiva were observed at 1, 2, 3, 4, and 7 days after administration of the test substance. Separately, after washing for 30 seconds using the three eye wash groups, the washing effect was confirmed, and the obtained results are shown in Table 8 below.

Ocular mucosa stimulation test results of the composition of Example 1 Number (mari) Eye irritation evaluation (M.I.O.I) Acute Eye Stimulation Index Day 2 3rd day Day 4 Day 5 8th day Bsean 6 0 0 0 0 0 0 Face wash 3 0 0 0 0 0 0

According to the evaluation of Draize's eye irritation, eye irritation of the test substance was scored, and the degree of the Eye Index of Ocular Irritation (M.I.O.I) was classified with reference to Guillot's method. As a result, eye lesions of the cornea, iris and conjunctiva were not found in all cases in the non-eye wash group at 1, 2, 3, 4 and 7 days after the administration of the test substance.

The index of Acute Ocular Irritation (I.A.O.I.) of the non-eyes group was determined to be '0' and 'non-irritant'. In the eye group, eye lesions of the cornea, iris and conjunctiva were not found in all cases at 1, 2, 3, 4 and 7 days after the administration of the test substance. No significant changes in general symptoms and no deaths were observed during the observation period, and no abnormal changes in body weight were observed.

From the above result, the composition of Example 1 which is a composition which concerns on this invention is judged to be a substance without irritation with respect to the ocular mucosa of a rabbit.

Experimental Example 10: Skin irritation test using rabbit

The skin irritation test of the composition of Example 1 as a test substance was carried out using six 16-week-old male NZW rabbits. Test substance was administered as is. In the skin of the depilated rabbit, two compartments of left and right, 2.5 × 2.5 cm ² , and a total of four compartments were set as administration sites. Of these, two compartments were rubbed and the other two were rubbed and sited, and 0.5 mL of the test stock solution was applied to the rubbed and unfriedged areas of each compartment. Only gauze was applied to the untreated control site, and the obtained results are shown in Table 9 below.

Skin irritation test results of the composition of Example 1 Mirror and site Abrasions 24 hours 48 hours 72 hours 24 hours 48 hours 72 hours Control 0 0 0 0 0 0 Test group 0 0 0 0 0 0

At 24, 48 and 72 hours after the administration of the test substance, the skin reaction was evaluated according to 'Draize Skin Response Evaluation Table 1)' to determine the skin irritation of the test substance. As a result, skin reactions such as erythema and edema at the site of administration of test substance at the 24, 48 and 72 hours after administration were not confirmed in all cases.

Skin reactions such as erythema and edema were not found in all cases. No mortality was observed during the observation period. There were no changes in general symptoms and abnormal body weights due to the administration of test substances. From the above result, the composition of Example 1 which is a test substance in this test condition is judged to be a substance without irritation to rabbit skin.

Experimental Example 11: Phototoxicity test using guinea pig

The phototoxicity test was performed using Hartley type male guinea pigs in order to examine the possibility of phototoxicity to the skin of the composition of Example 1 as a test substance. The test group was set to three groups of test group, control group 1 and control group 2 (5 mice each). As for the administration site, 1 site of left and right and 2 sites of sum were set symmetrically about the midline of the dorsal skin of the guinea pig. In the test group, 30% test substance was applied, control group 1 was injected with water for injection, and control group 2 (positive substance group) was 0.1 mL 8-methoxypsoralen (0.05 mL), respectively. UVA (320-400 nm) was irradiated so that the final energy might be about 10 J / cm ² with the light irradiation site on the left side and the non-light irradiation site on the right side. Skin irritation was determined at 24, 48 and 72 hours after light irradiation, phototoxicity was evaluated, and the results obtained are shown in Table 10 below.

Phototoxicity test results of the composition of Example 1 group time Non-light irradiation site Light irradiation site (Erythema total score + Edema total score) / N MTS (Erythema total score + Edema total score) / N MTS Test group 24 hours 48 hours 72 hours (0 + 0) / 5 (0 + 0) / 5 (0 + 0) / 5 0.0 0.0 0.0 (0 + 0) / 5 (0 + 0) / 5 (0 + 0) / 5 0.0 0.0 0.0 Control group 1 24 hours 48 hours 72 hours (0 + 0) / 5 (0 + 0) / 5 (0 + 0) / 5 0.0 0.0 0.0 (0 + 0) / 5 (0 + 0) / 5 (0 + 0) / 5 0.0 0.0 0.0 Control 2 24 hours 48 hours 72 hours (0 + 0) / 5 (0 + 0) / 5 (0 + 0) / 5 0.0 0.0 0.0 (15 + 20) / 5 (15 + 20) / 5 (15 + 20) / 5 7.0 7.0 7.0

Referring to Table 10, the test group and the control group at all observation time irrespective of the irradiation of the skin, such as erythema and edema was not observed in all cases. On the other hand, in the positive substance group, erythema of grade 3 and edema of grade 4 were observed in all the irradiated areas, but skin reactions such as erythema and edema were not observed in all the non-irradiated sites. There were no deaths during the observation period, and no effects of test substance administration were observed on changes in general symptoms and body weight. From the above result, the composition of Example 1 which is a test substance in this test condition is judged as a non-phototoxic substance.

In the above experimental results, the composition according to the present invention was found to be basically a safe component as a cosmetic composition because it is excellent in antimicrobial activity and not toxic.

Hereinafter, the composition of the cosmetic composition containing the plant extract prepared in Example 1-2 for Formulation Example 1-5 will be described in more detail. However, the composition of the present invention is not limited to these formulation examples.

Formulation Example 1 Softener (Skin Lotion)

Ingredient weight% Composition of Example 1 or 2 0.5 glycerin 3.0 Butylene glycol 2.0 Propylene glycol 2.0 Carboxy Vinyl Polymer 0.1 Fiji-12 nonylphenyl ether 0.2 Polysorbate 80 0.4 ethanol 10.0 Triethanolamine 0.1 Preservative, coloring, flavoring Quantity Purified water to 100

Formulation Example 2 Nutrients (Milk Lotion)

Ingredient weight% Composition of Example 1 or 2 0.5 Squalane 5.0 Beeswax 4.0 Polysorbate 60 1.5 Sorbitan sesquioleate 1.5 Liquid paraffin 0.5 Caprylic / Capric Triglycerides 5.0 glycerin 3.0 Butylene glycol 3.0 Propylene glycol 3.0 Carboxy Vinyl Polymer 0.1 Triethanolamine 0.2 Preservative, coloring, flavoring Quantity Purified water to 100

Formulation Example 3 Nutrition Cream

 Ingredient weight% Composition of Example 1 or 2 0.5 Beeswax 10.0 Polysorbate 60 1.5 Sebum 60 Cured Castor Oil 2.0 Sorbitan sesquioleate 0.5 Liquid paraffin 10.0 Squalane 5.0 Caprylic / Capric Triglycerides 5.0 glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Preservative, coloring, flavoring Quantity Purified water to 100

Formulation Example 4 Pack

Ingredient weight% Composition of Example 1 or 2 0.5 Polyvinyl alcohol 13.0 Sodium Carboxymethyl Cellulose 0.2 glycerin 5.0 Allantoin 0.1 ethanol 6.0 Fiji-12 nonylphenyl ether 0.3 Polysorbate 60 0.3 Preservative, coloring, flavoring Quantity Purified water to 100

Formulation Example 5 Gel

Ingredient weight% Composition of Example 1 or 2 0.5 Ethylenediamine Sodium Acetate 0.05 glycerin 5.0 Carboxy Vinyl Polymer 0.3 ethanol 5.0 Fiji-60 Cured Castor Oil 0.5 Triethanolamine 0.3 Preservative, coloring, flavoring Quantity Purified water to 100

1 is a photograph showing the antimicrobial activity of the extract prepared in Examples 1 and 2 and the control.

Claims (16)

Antimicrobial and antiseptic composition comprising nutmeg ( Myristica fragrans ), cloves ( Eugenia aromatica ), Eucalyptus alba , and Citrus paradisi extract. The method of claim 1, The extract is an antimicrobial and antiseptic composition that is extracted from the seeds of nutmeg, cloves seeds, eucalyptus leaves, grapefruit seeds. The method of claim 1, The extract is water; Alcohols of methanol, ethanol, isopropanol, or butanol; The antimicrobial and antiseptic composition obtained by using each of organic solvents of ethylene, acetone, ether, hexane, chloroform, or ethyl acetate or a mixed solvent thereof. The method of claim 1, The nutmeg, cloves, eucalyptus and grapefruit extract are 1 to 50% by weight of the antimicrobial and antiseptic composition, respectively, in the total composition. The method of claim 1, The extract is an antimicrobial and antiseptic composition comprising nutmeg, cloves, eucalyptus and grapefruit extract in a weight ratio (solid content) of 1: 2: 2: 1, respectively. The method of claim 1, In addition, the extract is antibacterial and antiseptic composition obtained by further performing a fermentation step using Bifidobacterium sp. , Lactobacillus sp. , Or a mixed strain thereof. The method of claim 1, The composition is Escherichia coli (KCTC 2571), Staphylococcus aureus KCTC 1916, Pseudomonas aeruginosa KCTC 2513, Candida albicans ( Cedda albicans KCTC 7965), and Aspergillus Aspergillus niger KCTC 6911, Propionibacterium acne, or An antimicrobial and antiseptic composition having antimicrobial activity against Ptyrosporum ovale . Cosmetics, food, pharmaceuticals, pesticides or household goods comprising the antimicrobial and antiseptic composition according to claim 1. Cosmetic composition having antimicrobial and antiseptic activity, including nutmeg ( Myristica fragrans ), cloves ( Eugenia aromatica ), Eucalyptus alba , and Citrus paradisi extract. The method of claim 9, The extract is a cosmetic composition that is extracted from the seeds of nutmeg, cloves seeds, eucalyptus leaves, grapefruit seeds. The method of claim 9, The extract is water; Alcohols of methanol, ethanol, isopropanol, or butanol; Organic solvents of ethylene, acetone, ether, hexane, chloroform, or ethyl acetate; Cosmetic composition obtained using each or these mixed solvents. The method of claim 9, The nutmeg, cloves, eucalyptus and grapefruit extract will each comprise 1 to 50% by weight in the total composition. The method of claim 9, The extract is nutmeg, cloves, eucalyptus and grapefruit extract each of the cosmetic composition comprising a weight ratio (solid content) of 1: 2: 2: 1. The method of claim 9, In addition, the extract is a cosmetic composition that is obtained by further performing a fermentation step using Bifidobacterium sp. , Lactobacillus sp. Strain or a mixed strain thereof. The method of claim 9, The composition is Escherichia coli (KCTC 2571), Staphylococcus aureus KCTC 1916, Pseudomonas aeruginosa KCTC 2513, Candida albicans ( Cedda albicans KCTC 7965), and Aspergillus Aspergillus niger KCTC 6911, Propionibacterium acne, or Cosmetic composition having an antimicrobial activity against Ptyrosporum ovale . The method of claim 9, The composition may be a flexible lotion, nourishing lotion, massage cream, nourishing cream, pack, gel, body lotion, ointment, gel, cream, patch, spray, shampoo, body shampoo, rinse, cleansing, face wash, soap, treatment, or essence Cosmetic composition is prepared in the formulation of.

Images