KR101532005B1 - Natural preservative comprising citrus junos seed extract and manufacturing method thereof - Google Patents

Abstract

The present invention relates to a natural preservative comprising a Citrus junos seed extract and a manufacturing method thereof. More specifically, the present invention can exhibit a wide range of antibacterial spectrums with respect to gram-positive bacteria, gram-negative bacteria, yeasts and fungi, an antiseptic ability, and a strong antioxidant effect by a natural preservative comprising a fermented Citrus junos seed extract, a Citrus junos seed extract, a scutellaria root extract, and an illicium verum Hooker extract. In addition, the present invention may have a skin care effect by a cosmetic composition containing the natural preservative.

Description

TECHNICAL FIELD [0001] The present invention relates to a natural preservative comprising a citrus peel extract, a citrus peel extract,

The present invention relates to a natural preservative comprising a citrus extract and a preparation method thereof, and more particularly to a natural preservative comprising a citrus extract, a citrus extract, a golden extract and an octagonal extract, , A broad spectrum antimicrobial spectrum against yeasts and fungi, a buoyant force and a strong antioxidant effect, and a process for producing the same.

In recent years, synthetic preservatives and synthetic additives such as parabens and phenoxyethanol, artificial fragrance and artificial coloring have been causing carcinogenesis in the human body and causing various skin irritation, so that demand for natural cosmetics such as no- .

The cosmetics industry is already using only paraben-free additives or selected preservatives, and natural preservatives are a preference. Since the early 2000s, the risk of parabens has been reported in the cosmetics industry. Developed countries, especially in Europe and North America, are developing alternatives to parabens and replacing them with natural preservatives in the long term.

Demand for antimicrobial products has increased dramatically. As of 2000, the global market size is estimated at about $ 3.4 billion, and the domestic market is estimated at 350 billion won, which is growing at an average annual rate of 15% or more (Source: The Daily Cosmetics ).

However, the development of such antibacterial materials in Korea is only a few companies, and most of them import raw materials from abroad. Currently, the most widely used natural preservative in the world market is the Grapefruits seed extract, DF-100, which is dominating the natural preservative market due to its lack of competing products.

The oldest grapefruit seed extract, which is the oldest natural preservative, is a natural preservative extracted from grapefruit peel and seeds. It is added to preservatives such as beverages, tofu, rice cakes and other cosmetics such as skin lotion, .

However, the above-mentioned grapefruit seed extract is used as a supplementary preservative rather than a preservative in the case of cosmetics because the stability of the product is lowered, the range of preservation spectrum is narrow and the preservation endurance is decreased. In addition, since the grapefruit itself is not produced commercially in Korea, the grapefruit seed extract has a problem in that it is expensive and expensive.

On the other hand, Patent Document 1 discloses a cosmetic composition containing a natural extract and a method for producing the same, but has limitations in showing a wide range of antibacterial activity against various microorganisms.

Patent Document 1: Korean Patent Publication No. 2003-0039839

One aspect of the present invention is to provide a natural preservative comprising a fermented extract of Yuzu seed.

Another aspect of the present invention is to provide a cosmetic composition comprising the natural preservative.

Another aspect of the present invention is to provide a method for producing a natural preservative containing the fermented Yuza seed extract.

A natural preservative according to an embodiment of the present invention comprises: 1 to 60% by weight of a fermented extract of citrus seed; 1 to 30% by weight citrus seed extract; 1 to 80% by weight of a gold extract; And 1 to 60% by weight of octagonal fennel extract.

In the natural preservative, the citrus fruit fermentation extract may be produced by fermenting citrus seed with lactic acid bacteria.

In the natural preservative, the lactic acid bacteria may be selected from the group consisting of Lactobacillus brevis, Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus plantarum, Lactobacillus kimchi Lactobacillus < / RTI > kimchii, Lactobacillus paraplantarum, Lactobacillus carbatus subcipy. Lactobacillus curvatus subsp. Curvatus, and Lactobacillus subtilis. One or more can be selected from Lactobacillus sakei subsp. Sakei.

The cosmetic composition according to one embodiment of the present invention may contain 0.1 to 30% by weight of the natural preservative.

The cosmetic composition may be a single formulation selected from a cream, a skin lotion, an essence, a foundation, a hair shampoo, a hair treatment, a hair tonic and a hair preparation.

A method for preparing a natural preservative according to one embodiment of the present invention comprises:

(A) mixing the citron seeds in a culture medium for lactic acid bacteria and fermenting them to obtain a fermented citrus fruit extract;

(B) obtaining an extract of citrus peel from citron;

(C) obtaining a gold extract from gold;

(D) obtaining an octopus fennel extract from octopus fennel; And

(E) mixing 1 to 60% by weight of the above fermented citrus fruit extract, 1 to 30% by weight of citrus seed extract, 1 to 80% by weight of a golden extract and 1 to 60% by weight of an octagonal fennel extract.

Wherein the lactic acid bacterium is selected from the group consisting of Lactobacillus brevis, Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus plantarum, Lactobacillus spp. Lactobacillus kimchii, Lactobacillus paraplantarum, Lactobacillus carbatus subcipy. Lactobacillus curvatus subsp. Curvatus, and Lactobacillus subtilis. One or more can be selected from Lactobacillus sakei subsp. Sakei.

The natural preservative according to one embodiment of the present invention can exhibit a wide range of antimicrobial activity, antiseptic and antioxidative effects against Gram-positive bacteria, Gram-negative bacteria, yeast and fungi. In addition, the cosmetic composition according to one embodiment of the present invention can exhibit a wide range of antimicrobial activity, antiseptic and antioxidative effects against Gram-positive bacteria, Gram-negative bacteria, yeast and fungi. In addition, the natural preservative containing the citron extract according to one embodiment of the present invention and the preparation method thereof can be expected to have a skin cosmetic effect through a broad spectrum of antimicrobial spectrum, preservation and strong antioxidant of the natural extract.

FIG. 1 is a photograph showing the anti-microbial activity of Staphylococcus aureus prepared according to Example 4 of the present invention.
2 is a photograph showing the antimicrobial activity of a natural preservative prepared according to Example 1 of the present invention against E. coli;
3 is a photograph showing the antimicrobial activity of the natural preservative prepared according to Example 4 of the present invention against black germ.
FIG. 4 is a graph showing the cytotoxicity of the Yuza seed fermented extract prepared through Production Example 1 of the present invention by MTT test; And
FIG. 5 is a graph showing the cytotoxicity of the yuza seed extract prepared through Production Example 2 of the present invention by MTT test.

Before describing the invention in more detail, it is to be understood that the words or words used in the specification and claims are not to be construed in a conventional or dictionary sense, It should be interpreted as meaning and concept consistent with the technical idea of the present invention. Therefore, the constitution of the embodiments described in the present specification is merely a preferred example of the present invention, and does not represent all the technical ideas of the present invention, so that various equivalents and variations And the like.

Hereinafter, preferred embodiments of the present invention will be described in detail so that those skilled in the art can easily carry out the present invention. In the following description, well-known functions or constructions are not described in detail since they would obscure the invention in unnecessary detail.

Natural preservative

The natural preservative according to one embodiment of the present invention includes a fermented extract of Yuzu seed, a yuza seed extract, a golden extract, and an octagonal fennel extract.

In the present invention, the term "natural preservative" refers to a preservative having a broader antimicrobial spectrum due to the combined action of two or more substances extracted from a natural substance in general. However, It can have the same meaning as antimicrobial agent. Therefore, in the present invention, the natural preservative may also be used to mean preservative function and antibacterial function.

The natural preservative of the present invention is not only toxic and irritating to human body but also safe for use in the human body as it not only has excellent antimicrobial activity, but also can be used for cosmetics, medicines, foods and the like.

The above-mentioned antimicrobial spectrum can be a standard for indicating to which microorganism an antimicrobial substance such as an antimicrobial substance or a synthetic antimicrobial substance is effective. The antimicrobial activity is expressed as the minimum concentration (Minimal Inhibitory Concentration, MIC) of the antimicrobial substance that can not grow on the plate of the medium containing the agar. The antimicrobial spectrum is a list of MICs of antimicrobial substances against various pathogens and is a criterion for judging which antimicrobial substance is effective for infection of a microorganism when treating an infectious disease patient. Depending on the antimicrobial substance, it is used for the treatment of many infectious diseases because it has a broad range of bacteria (broad antimicrobial substance). Antimicrobial substances with spectrum up to fungi such as yeast and fungi are often strong against animal cells and often can not be used for treatment.

Citrus junos is a wild plant which is cultivated in the southern part of Korea. It is a botanical fruit, a citrus fruit, and a citrus fruit which belongs to the genus Citrus. It is strong in water and strong in standing, and is resistant to cold tolerance and tolerance.

Yuzu, which is the fruit of citron, is widely used for ornamental, edible, medicinal, and spice, and the fruit shell is called citron, flour,

The radionuclide contains a fatty oil, a protein, and a limonoid component as a gummy ingredient. The limonoid is one of the triterpene derivatives, and has a carcinostatic action, an insect's food-feeding inhibiting function, and the like.

Limonoids detected in citron are Diacetylnomillin, Nomillin, Limonin and Ichanginsin, and the content is the most in the seed rather than the perilla or flesh.

The limonoid is a component showing the bitter taste of citron, and it is known that it has an anticancer effect medically though it is an obstacle factor in processing (source: National Species Information System, Jeollanamdo Agricultural Technology Institute).

The fermented extract of Yuzu seeds according to one embodiment of the present invention can be obtained by fermenting and extracting waste seeds that have been discarded in the processing of yuzhou processing using raw materials.

The fermented extract of Yuzu seeds is prepared by inoculating a fermentation strain with Yuza seed to produce a new physiologically active substance in the course of decomposition of the organic matter by the enzyme of the strain and to increase the content of the nutrients, It can have the effect of reducing side effects.

The fermented extract of Yuzu seeds can increase antimicrobial activity through bio-conversion using lactic acid bacteria, and has low cytotoxicity and thus is excellent in safety.

In the natural preservative, the citrus fruit fermentation extract may be produced by fermenting citrus seed with lactic acid bacteria.

Here, the fermentation extract can be obtained by filtering a fermentation broth obtained by fermenting citron seeds with lactic acid bacteria in a medium inoculated with lactic acid bacteria. Accordingly, the natural preservative according to one embodiment of the present invention may include a fermentation broth produced by mixing citron seeds with a culture medium for lactic acid bacteria and inoculating and fermenting the lactic acid bacteria.

The lactic acid bacteria may be selected from the group consisting of Lactobacillus brevis, Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus plantarum, Lactobacillus kimchii, Lactobacillus paraplantarum, Lactobacillus carbatus subcapsule. Lactobacillus curvatus subsp. Curvatus, and Lactobacillus subtilis. Lactobacillus sakei subsp. Sakei may be selected from one or more, but not limited thereto.

The natural preservative may have antimicrobial activity from one or more selected strains of Gram-positive bacteria, Gram-negative bacteria, yeast, and fungi. Specifically, for example, the strain may include Staphylococcus aureus, Escherichia coli, Candida albicans, Aspergillus niger, and the like. And is not particularly limited thereto.

The fermented citrus fruit extract according to one embodiment of the present invention may be contained in the natural preservative in an amount of 1 to 60% by weight. If the yuzu seed fermented extract is less than 1% by weight, the preservative effect and efficacy tend to be weak. If the yuzu seed fermented extract is more than 60% by weight, the stabilization of the cosmetic product may be affected.

The citron extract according to one embodiment of the present invention has an excellent repelling ability against E. coli and can assist the fermented extract of citron juice having a weak antibacterial effect against E. coli by biotransformation.

The citron extract may be at least one selected from the group consisting of water, methanol, ethanol, propanol and butanol, acetone, ethyl acetate, chloroform, butyl acetate and 1,3-butylene glycol.

The citron extract according to one embodiment of the present invention may contain 1 to 30% by weight of the natural preservative. If the yuzu seed extract is less than 1% by weight, there may be no flotation. If the content is more than 30% by weight, the flotation does not affect the increase of the flotation power, but the stability may be deteriorated.

Golden (Scutellariae radix) belongs to the family Lamiaceae. It grows in various parts of Korea and contains Baicalin, Baicalein and Woogonin to suppress allergic edema and inflammation, It has an inhibitory effect, and it has fever, diuretic, asthma, and anti-cancer action. The gold active material is typically known as baicalein.

The gold extract may be at least one selected from water, methanol, ethanol, propanol and butanol, acetone, ethyl acetate, chloroform, butyl acetate and 1,3-butylene glycol in the extraction process.

The gold extract may comprise from 1 to 80% by weight of the natural preservative. If the amount of the gold extract is less than 1% by weight, there may be no repellency. If the amount of the extract is more than 80% by weight, it may not affect the repulsive force.

Octagonal fennel is a fruit of star anise (Illicium verum) which warms the Yang, releases the cold and regulates the circulation of the ki, It is known as an effective medicinal herbal medicine for the treatment of abdominal pain caused by swelling and backache, cold pain caused by abdominal pain caused by the kidneys, and dryness or dryness.

The octagonal fennel has antimicrobial activity against bacteria such as Staphylococcus aureus, Escherichia coli and fungus, and can enhance the antifungal effect when it is combined with a plant extract.

The octagonal fennel extract may be one or more selected from water, methanol, ethanol, propanol and butanol, acetone, ethyl acetate, chloroform, butyl acetate and 1,3-butylene glycol in the extraction process.

The octopus fennel extract may contain 1 to 60% by weight of the natural preservative. If the octagonal fennel extract is less than 1% by weight, it may be free of flotation, and if it exceeds 60% by weight, it may not stimulate the increase of flotation power, but it may irritate the skin.

In addition, the natural preservative composition according to one embodiment of the present invention may further include a useful vegetable antibacterial extract including a grapefruit extract and a microbial culture extract, and is not particularly limited thereto.

In addition, the natural preservative composition according to the present invention can be used in the form of a spray or the like, exhibits a deodorizing effect and a cleaning effect by an antibacterial function, and can be used as a deodorant or a detergent if necessary.

Cosmetics  Composition

The cosmetic composition according to one embodiment of the present invention may contain a natural preservative as described above.

The cosmetic composition according to one embodiment of the present invention may contain 0.1 to 30% by weight of a natural preservative. If the amount of the natural preservative is less than 0.1 wt%, the repellency and antioxidant function of the cosmetic product may be weak. If the natural preservative is more than 30 wt%, the content of the other composition may be decreased.

The cosmetic composition may have one formulation selected from a cream, a skin lotion, an essence, a foundation, a hair shampoo, a hair treatment, a hair tonic, and a hair cosmetic composition.

The cosmetic composition according to one embodiment of the present invention can be used in cosmetics such as purified water, humectants, high alcohol, higher fatty acid, emollient, chelating agent, thickener, nutritional ingredient, pH regulator, . ≪ / RTI >

The cosmetic composition according to one embodiment of the present invention may contain antibacterial, antiseptic and antioxidative effects as compared with conventional cosmetic compositions by containing a natural preservative including a fermented extract of Yuzu seed.

Preservatives which have been used for the purpose of inhibiting the growth of microorganisms capable of contaminating cosmetics from the outside and used for the purpose of preventing the change of products, for example, synthetic compounds such as paraoxybenzoic acid ester, methylparaben, ethylparaben, propylparaben, Preservatives have problems of skin allergies, environmental hormones and inducing resistant bacteria.

However, the cosmetic composition according to one embodiment of the present invention can exhibit a preservative effect even in a small amount by replacing the synthetic preservative, exhibits high antibacterial activity and high antibacterial activity against resistant bacteria and fungus without inducing resistance, By including the fermented extract of Yuzu seeds showing the spectrum, it is possible to exhibit an excellent safety and economical effect as compared with the conventional cosmetic composition.

Method of manufacturing natural preservative

A method for preparing a natural preservative according to one embodiment of the present invention comprises:

(A) mixing the citron seeds in a culture medium for lactic acid bacteria and fermenting them to obtain a fermented citrus fruit extract;

(B) obtaining an extract of citrus peel from citron;

(C) obtaining a gold extract from gold;

(D) obtaining an octopus fennel extract from octopus fennel; And

(E) mixing 1 to 60% by weight of the above fermented citrus fruit extract, 1 to 30% by weight of citrus seed extract, 1 to 80% by weight of a golden extract and 1 to 60% by weight of an octagonal fennel extract.

The lactic acid bacteria may be selected from the group consisting of Lactobacillus brevis, Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus plantarum, Lactobacillus kimchii, Lactobacillus paraplantarum, Lactobacillus carbatus subcapsule. Lactobacillus curvatus subsp. Curvatus, and Lactobacillus subtilis. One or more can be selected from Lactobacillus sakei subsp. Sakei.

The fermentation may be performed using a lactic acid bacterium concentrated at a temperature of 20 to 40 DEG C, 24 to 120 hours, and 10 ⁵ to 10 ⁹ CFU / mL.

According to one embodiment of the present invention, the extracts of yuzu, golden, and octagonal fennel are prepared by mixing water, methanol, ethanol, propanol, butanol, acetone, ethyl acetate, chloroform, butyl acetate, 3-butylene glycol, and the like. The extraction is carried out at a temperature between 20 and 80 ° C. for 6 to 72 hours, followed by filtration through a filter paper having a pore size of 0.45 μm and concentration under reduced pressure.

The cultivation and fermentation of the lactic acid bacteria according to one embodiment of the present invention can be performed according to a suitable culture medium and culture conditions known in the art. Such a process can be easily adjusted according to the strain selected by those skilled in the art.

Further, after the completion of the culture, centrifugation or filtration may be carried out to separate the culture from the cells, and may be appropriately carried out by a person skilled in the art if necessary. At this time, the removal of the cells indicates that the lactic acid bacterium cells have been substantially removed through the conventional method for removing the cells.

Hereinafter, the present invention will be described in more detail with reference to Examples and Comparative Examples. However, the scope of the present invention is not limited to the following examples.

Production Example 1. Preparation of Yuza seed fermented extract

The seeds of citron were dried and sterilized by heating at 121 ° C for 15 minutes. After sterilization, the mixture was fermented at 35 DEG C for 50 hours using Lactobacillus brevis. After the fermentation was completed, the fermentation was sterilized to kill the fermentation strain, and the fermented Yuzu seed extract was extracted from the fermented Yuzu seeds with ethanol. The fermented Yuzu seed extract was prepared by filtration through a filter having a thickness of 0.2 탆 so as to prevent the formation of a precipitate.

Production Example 2. Preparation of yuza seed extract

The commercially available citron was purchased and immersed in ethanol, a solvent, and extracted for about 48 hours. Then, the extract which was filtered with a filter having a thickness of 0.2 탆 was concentrated under reduced pressure at about 45 캜 to prepare dried yuza seed extract.

Production Example 3. Preparation of golden extract

Commercially available gold was purchased and immersed in ethanol, a solvent, and extracted for about 48 hours. Then, the extract which had been filtered with a filter having a thickness of 0.2 탆 was concentrated under reduced pressure at about 45 캜 to prepare a dried gold extract.

Production Example 4 Octagonal fennel  Preparation of extract

Commercial octagonal fennel was purchased and immersed in ethanol as a solvent and extracted for about 48 hours. Then, the extract which was filtered with a filter having a thickness of 0.2 탆 was concentrated under reduced pressure at about 45 캜 to prepare dried octagonal fennel extract.

Measurement of antimicrobial activity of extract

Experiments were conducted to measure the antimicrobial activity of the fermented Yuza seed extract, Yuza seed extract, golden extract and octopus fennel extract of Production Examples 1 to 4.

Staphylococcus aureus as gram-positive bacteria, Escherichia coli as Gram-negative bacteria, Candida albicans as yeast, and fungi as yeast were used as the measurement methods. Aspergillus niger was used. Approximately 100 μl of each of the cultivated strains was dispensed on a solidified plate medium, dispensed in a plate culture medium, and uniformly smoothed using a spreader.

Each of the extracts prepared in Preparation Examples 1 to 4 was absorbed in an amount of about 25 μl into a circular filter paper having a diameter of about 8 mm, followed by ethanol drying, and the strain was brought into close contact with the surface of the medium After culturing for 24 hours or more in an incubator under a temperature condition, the size of the clear zone formed around the circular filter paper was measured to confirm the antibacterial effect.

division Bacterium Staphylococcus aureus Escherichia coli Candida yeast Black germ Production Example 1 +++ - +++ ++ Production Example 2 + +++ - + Production Example 3 ++ + +++ ++ Production Example 4 ++ ++ - +

※ Table 1 indicates the criteria for antimicrobial activity

① (-): No effect

② (+): Normal

③ (++): Good

④ (+++): Very good

As can be seen from the above Table 1, the antibacterial activity test (circular filter paper method) of the Yuza seed fermentation extract, Yuza seed extract, Golden extract and Octagonal Fennel extract prepared in Production Examples 1 to 4 was as follows. Indicating that it exhibits excellent activity against staphylococci. In the case of Escherichia coli, the antimicrobial activity of the three plant extracts except the fermented Yuzu seed extract was exhibited, and the Candida yeast showed excellent antimicrobial activity in fermented Yuza seed extract and gold extract. Black germ extract showed good activity in fermented and fermented extracts of yuza seeds and weak activity in the other two extracts.

Microbial minimum inhibitory concentration measurement

Yuza seed extract, Yuza seed extract, Golden extract, and Octagonal fennel extract were evaluated for their antibacterial activity against the strain.

Staphylococcus aureus, Escherichia coli, Candida albicans, and Aspergillus niger were used as microorganism strains.

Staphylococcus aureus and E. coli were preincubated for 24 hours at 37 ° C in Brain Heart Infusion Agar and diluted to a concentration of 10 ^5-6 CFU / ml to prepare the inoculum.

Candida yeast was preincubated in a potato dextrose agar for 48 hours at a temperature of 30 ° C to prepare 10 ^5-6 CFU / ml inoculum.

Black seed germs were inoculated on a slab medium (Potato Dextrose Slant media) made of solid medium for mold and preincubated at 30 ° C for 48 hours to prepare a broth at a concentration of ^5-6 CFU / ml.

Brain Heart Infusion Broth was used for Staphylococcus aureus and Escherichia coli, and Potato Dextrose Broth was used as a liquid medium for Candida yeast and Black germ.

100 μl of the liquid medium prepared as described above was added to each well of a 96-well plate, and the sample diluted solution and the microorganism were inoculated. The staphylococcus aureus and E. coli were inoculated in a 37 ° C. incubator for 24 hours, Was incubated for 48 hours in an incubator at a temperature of 30 ° C, and the degree of growth was measured at 600 nm using an ELISA reader.

The lowest concentration of the sample in which no growth of the microorganism was determined was determined as the Minimal Inhibitory Concentration (MIC) of the antimicrobial substance against the microorganism.

division Bacterium Staphylococcus aureus Escherichia coli Candida yeast Black germ Production Example 1 0.03% - 0.03% 0.06% Production Example 2 1.0% 0.03% - 2.4% Production Example 3 0.5% 1.5% 0.03% 1.0% Production Example 4 0.25% 0.06% - 0.5%

As shown in Table 2, the minimum growth inhibitory concentration (MIC) of each of the extracts prepared in Preparation Examples 1 to 4 is shown.

In the case of the preparation example 1, it can be seen that the antimicrobial activity is excellent at a concentration of 0.03 to 0.06% with respect to all bacteria except for Escherichia coli.

In the case of the preparation example 2, it can be seen that the antimicrobial activity is excellent in E. coli, but the antimicrobial activity is relatively weak against Staphylococcus aureus, Candida yeast and Black germ.

In the case of Preparation Example 3, it exhibited a weak antimicrobial activity against Escherichia coli and black microorganism, but it exhibited antibacterial activity effectively at a concentration of 0.03% for yeast as a fungus.

In the case of Preparation Example 4, it was found that the antibacterial activity was excellent at 0.25% for E. coli and 0.06% for E. coli, and 0.5% for black microorganism.

Manufacture of natural preservatives

Compositions mixed at various ratios were prepared in order to confirm the preservative effect of the natural preservative according to one embodiment of the present invention.

Examples 1 to 4

The natural preservative compositions of Examples 1 to 4 were prepared by mixing the respective extracts prepared in Preparation Examples 1 to 4 in the proportions shown in Table 3 below.

division Citron fermentation product Yuza seed extract Golden extract Octagonal fennel extract Example 1 40 wt% 20 wt% 20 wt% 20 wt% Example 2 10 wt% 20 wt% 20 wt% 50 wt% Example 3 40 wt% 30 wt% 20 wt% 10 wt% Example 4 40 wt% 10 wt% 30 wt% 20 wt%

Antimicrobial activity measurement of natural preservative

In order to evaluate the antimicrobial activity of the natural preservative against the strain, the experiment was carried out in the same manner as that of the antimicrobial activity of the above Preparation Examples 1 to 4.

division Bacterium Staphylococcus aureus Escherichia coli Candida yeast Black germ Example 1 0.12 0.25 0.5 0.5 Example 2 0.25 0.25 0.25 1.0 Example 3 0.06 0.12 0.25 1.0 Example 4 0.06 0.25 0.12 0.5

As can be seen from the above Table 4, the natural preservatives prepared in Examples 1 to 4 are superior to the respective extracts prepared in Preparation Examples 1 to 4 in the presence of bacteria (Staphylococcus aureus and Escherichia coli) and fungi It was found that the antibacterial activity of the natural extract containing yuza seed showed an excellent antimicrobial activity as a whole, and in particular, the microbial minimum inhibitory concentration of the natural preservative prepared in Example 4 And the antimicrobial activity is the most excellent.

Antimicrobial activity measurement of natural preservative

In order to measure the antimicrobial activity of the natural preservative prepared in Example 1, the FITI test institute commissioned an antibacterial activity test. The natural preservative composition of Example 1 was diluted to a concentration of 2%. Staphylococcus aureus was used as the strain used, and Escherichia coli was used as gram-negative bacteria.

Staphylococcus aureus and Escherichia coli with an initial number of bacteria of 1.4 x 10 ⁴ bacteria / ml were inoculated into Example 4 (a natural preservative containing yuzu seed) diluted to a concentration of 2%, followed by shaking culture at a temperature of 37 캜 for about 24 hours As a result, 99.9% of bacteria were decreased in both Staphylococcus aureus and Escherichia coli (see FIGS. 1 to 2).

After wetting Example 4 (a natural preservative containing yuzu seed) diluted to a concentration of 2% with black bacillus at an initial number of bacteria of 1.0 占 104 bacterium / ml, the mixture was cultured at a temperature of 24 占 폚 for 7 days, As a result, it was confirmed that black microbes did not grow (see Fig. 3).

Cytotoxicity test

In order to verify the primary safety, the MTF test was carried out by culturing the HDF-cells (human fibroblasts) on the fermented YUJA fermentation extract and YUJA seed extract prepared in Preparation Examples 1 and 2 to evaluate cytotoxicity The results are shown in FIGS. 3 and 4, respectively. The cell viability (%) was compared with the control group, DPBS (untreated group) after treating the fermented extract of Yuzu seed (Preparation Example 1) and yuza seed extract (Preparation Example 2) at a concentration of 0.05 to 2%.

It was confirmed that the cell proliferation efficacy was maintained when the fermented extract of Yuzu seed and the extract of Yuza seed were treated with HDF-cells at a concentration as compared with the control, and it can be safely used in a cosmetic composition.

Primary irritation test for human body ( A patch test )

In order to examine the safety of a sample containing a fermented extract of Yuzu seed and citron extract, a first stimulation test was performed on a human body through a human patch test as described below.

Ten healthy adult men and women were enrolled according to the CTFA guidelines (The Cosmetic, toiletry and Fragrance Association, Inc., Washington, D.C., 20046, 1991).

15 mg of the sample and 15 mu l of the solution were dropped into the pin chamber, and this was put on the arm of the human body to be tested and fixed with a tape. Then, the skin was reacted for 24 hours, and then the patch was removed. After 2 hours, the skin reaction of the test site was judged according to the following criteria.

reaction score Symptoms and judging criteria - 0 No response +/- 0.5 Weak erythema + One Clear erythema ++ 2 Severe erythema with edema +++ 3 Severe erythema, blisters, edema

Table 5 above is a table relating to the criteria.

sample Embroidery (person) Skin irritation index Preservative-free additive products 10 0.0 Production Example 1 (Fermented Yuzu seed extract at a concentration of 2%) [ 10 0.0 Production Example 2 (2% yuza seed extract) 10 0.0

※ Skin irritation index: [Σ (number of points × number of irritations)] / total number of test × number of total irritation evaluations

As can be seen from Table 6, the fermented Yuza seed extract and yuza seed extract, which are effective ingredients of the natural preservative according to one embodiment of the present invention, are found to be a skin-stable ingredient.

Manufacture of cream

Comparative Example 1

As shown in Table 7 below, a cream containing phenoxyethanol, a commonly used synthetic preservative, was prepared.

Examples 5 to 7

As shown in Table 7 below, a cream comprising a natural preservative according to one embodiment of the present invention was prepared.

Raw material name Comparative Example 1 Example 5 Example 6 Example 7 weight% weight% weight% weight% Cetostearyl alcohol 3.0 3.0 3.0 3.0 Self emulsifying monostearic acid 1.5 1.5 1.5 1.5 Chin type monostearate 1.5 1.5 1.5 1.5 Wax 0.5 0.5 0.5 0.5 Liquid paraffin 6.0 6.0 6.0 6.0 Squalane 8.0 8.0 8.0 8.0 Isocetyl octanoate 4.0 4.0 4.0 4.0 Refined jojoba oil 4.0 4.0 4.0 4.0 Dimethylsiloxane 0.3 0.3 0.3 0.3 Sorbitan monostearate 1.0 1.0 1.0 1.0 Polyethylene glycol monostearate 1.2 1.2 1.2 1.2 glycerin 8.0 8.0 8.0 8.0 Propylene glycol 4.0 4.0 4.0 4.0 Betaine 4.0 4.0 4.0 4.0 Xanthan gum 0.06 0.06 0.06 0.06 Triethanolamine 0.1 0.1 0.1 0.1 Spices Suitable amount Suitable amount Suitable amount Suitable amount Pigment Suitable amount Suitable amount Suitable amount Suitable amount Citron fermented product and extract - 2.0 - - Yuza seed extract - - 2.0 - Complex natural preservative - - - 2.0 Phenoxyethanol 0.7 - - - Purified water to 100 to 100 to 100 to 100

Buoyancy  Measure

As shown in Table 7, the cream containing the natural preservative containing the fermented extract of Yuzu seed was prepared and the buoyancy was measured. The cream containing phenoxyethanol as the general synthetic preservative was prepared in Examples 5 to 7, and the cream containing Comparative Example 1, Yuza seed fermented extract, Yuza seed extract and complex natural antibacterial agent.

The microbial strain used in the buoyancy measurement experiment is the same as the strain used for the above-mentioned antimicrobial activity measurement. The experiment was carried out in accordance with the CTFA Microbiology Guideline of the CTFA. Bacteria (Staphylococcus aureus and Escherichia coli) were inoculated at a concentration of 10 ⁶ CFU / ml, fungi (Candida yeast and black germ cells) were inoculated at a concentration of 10 ⁵ CFU / ml The number of bacteria was measured from 1 to 28 days after inoculation.

Due to the effectiveness of the buoyant force, bacteria should be reduced by 99.9% or more within 7 days of inoculation and there should be no proliferation during the test period. Yeast and fungi (specifically, whether they are related to the strain) should be reduced by at least 90% within 7 days of inoculation and not be proliferated during the test period.

division Number of bacteria according to the elapsed days The first number of bacteria 1st day tea 7th day car 14th day car 28th day car Staphylococcus aureus

Comparative Example 1 1.0 × 10 ⁶ <100 <20 <20 <20 Example 5 1.0 × 10 ⁶ 4.0 × 10 ⁴ <20 <20 <20 Example 6 1.0 × 10 ⁶ 2.0 × 10 ⁵ 1.6 x 10 ³ <100 <100 Example 7 1.0 × 10 ⁶ 1.0 x 10 ⁴ <20 <20 <20 Large intestine

Comparative Example 1 1.0 × 10 ⁶ <100 <20 <20 <20 Example 5 1.0 × 10 ⁶ 4.0 × 10 ⁵ <20 <20 <20 Example 6 1.0 × 10 ⁶ 2.0 × 10 ⁵ <20 <20 <20 Example 7 1.0 × 10 ⁶ 6.0 x 10 ³ <20 <20 <20 Candida
leaven

Comparative Example 1 1.0 × 10 ⁵ <100 <20 <20 <20 Example 5 1.0 × 10 ⁵ 1.0 × 10 ⁵ <100 <20 <20 Example 6 1.0 × 10 ⁵ 4.0 × 10 ⁵ 4.0 × 10 ⁵ 4.0 × 10 ⁵ 4.0 × 10 ⁵ Example 7 1.0 × 10 ⁵ 1.3 x 10 ⁴ <20 <20 <20 Black fungus

Comparative Example 1 1.0 × 10 ⁵ 1.0 x 10 ⁴ <20 <20 <20 Example 5 1.0 × 10 ⁵ 5.0 × 10 ⁴ <100 <20 <20 Example 6 1.0 × 10 ⁵ 5.0 × 10 ⁴ 1.5 x 10 ³ 1.0 x 10 ³ 1.0 x 10 ³ Example 7 1.0 × 10 ⁵ 2.0 x 10 ⁴ <20 <20 <20

As can be seen from Table 8, it can be seen that the cosmetic composition containing the natural preservative of the present invention exerts the same level of repelling power as the cosmetic composition using the conventional synthetic preservative.

In addition, the cosmetic composition prepared in Example 7, which contains both the yuza seed fermented extract, the yuza seed extract, the golden extract, and the octagonal fennel extract, has a better preservative performance than the yuza seed fermentation extract and the yuza seed extract Can be improved. Accordingly, the natural preservative according to one embodiment of the present invention can be usefully used in various cosmetic formulations for preservation purposes.

Manufacture of skin lotion

Comparative Example 2

As shown in Table 9 below, skin lotions containing phenoxyethanol, a commonly used synthetic preservative, were prepared.

Example 8

As shown in Table 9 below, a skin lotion comprising a natural preservative according to one embodiment of the present invention was prepared.

A skin containing 1% by weight of a natural preservative (Example 5) containing a fermented extract of Yuzu seed and an extract of Yuzu seed was prepared.

As a result of the preservative effect test in the same manner as that of measuring the repellency of the cream, 99.9% of the bacteria died before 7 days of bacteria and fungi in Comparative Examples 2 and 8, and 28 days The growth of the microorganisms was not observed even after the elapsed time. As a result, it was found that natural preservatives can be used for the skin formulations.

Raw material name Comparative Example 2 Example 8 weight% weight% Glycerin 3.0 3.0 1,3 BG 5.0 5.0 Sodium Hyaluronate 1% 1.5 1.5 Pine leaf extract 0.5 0.5 Ethanol 8.0 8.0 Spices Suitable amount Suitable amount Pigment Suitable amount Suitable amount Natural preservative (Example 4) - 1.0 Phenoxyethanol 0.7 - Purified water to 100 to 100

Manufacture of essences

Comparative Example 3

As shown in Table 10 below, essences containing phenoxyethanol, a commonly used synthetic preservative, were prepared.

Example 9

As shown in Table 10 below, an essence containing a natural preservative according to one embodiment of the present invention was prepared.

An essence containing 1% by weight of a natural preservative containing a fermented extract of Yuzu seed and an extract of Yuzu seed was prepared. As a result of the preservative effect test in the same manner as that of measuring the repellency of the cream, 99.9% of the bacteria were killed before 7 days of bacteria and fungi in Comparative Examples 3 and 9, and 28 days The growth of the microorganisms was not observed even after the elapsing. As a result, it can be seen that natural preservatives can be used for essence formulations.

Raw material name Comparative Example 3 Example 9 weight% weight% Sodium Hyaluronate 1% 3.0 3.0 1,3 BG 5.0 5.0 Glycerin 5.0 5.0 Location of Hazel extract 0.5 0.5 Hydrogenated Lecithin 0.2 0.2 Allantoin 0.1 0.1 Ethanol 95% 5.0 5.0 Hydrogenated casto oil (PEG 60) 0.5 0.5 Spices 0.1 0.1 Natural Betain 1.5 1.5 Xanthan gum 0.1 0.1 Pigment Suitable amount Suitable amount Natural preservative (Example 4) - 2.0 Phenoxyethanol 0.7 - Purified water to 100 to 100

Preparation of antibacterial deodorant

Example 10

As shown in Table 11 below, an antibacterial deodorant containing the fermented Yuzu seed extract according to an embodiment of the present invention was prepared.

Example 11

As shown in the following Table 11, an antimicrobial deodorant containing the natural preservative prepared in Example 4 of the present invention was prepared.

Antibacterial deodorant containing 1% by weight of natural preservative containing fermented Yuza seed extract and citron extract was prepared as shown in Table 11 below.

The natural preservative containing yuza seed fermentation extract and citron seeds is effective for deodorization and antibacterial action when it contains 0.01 to 30% by weight, preferably 1 to 10% by weight, and can be used as a deodorant for household antibacterial.

Raw material name Example 10 Example 11 weight% weight% ethanol 10.0 20.0 1,3 BG 3.0 3.0 Citric acid 0.1 0.1 Pine leaf extract 10.0 - Linseed oil - 10.0 Yuza seed fermentation extract 3.0 - Natural preservative (Example 4) - 3.0 Acetic acid tocopherol 0.1 0.1 Purified water to 100 to 100

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, the same is by way of illustration and example only and is not to be construed as limiting the present invention. It is obvious that it can be modified or improved.

It is intended that the present invention cover the modifications and variations of this invention provided they come within the scope of the appended claims and their equivalents.

Claims (7)

1 to 60% by weight of fermented Yuzu seed extract; 1 to 30% by weight citrus seed extract; 1 to 80% by weight of a gold extract; And 1 to 60% by weight of an octagonal fennel extract. The method according to claim 1,
The above fermented Yuzu seed extract is a natural preservative produced by fermenting Yuza seed with lactic acid bacteria. The method of claim 2,
The lactic acid bacteria may be selected from the group consisting of Lactobacillus brevis, Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus plantarum, Lactobacillus kimchii, Lactobacillus paraplantarum, Lactobacillus carbatus subcapsule. Lactobacillus curvatus subsp. Curvatus, and Lactobacillus subtilis. One or more natural preservatives selected from Lactobacillus sakei subsp. Sakei. A cosmetic composition comprising 0.1 to 30% by weight of a natural preservative according to claim 1. The method of claim 4,
Wherein the composition is one form selected from a cream, a skin lotion, an essence, a foundation, a hair shampoo, a hair treatment, a hair tonic, and a hair care composition. (A) mixing the citron seeds in a culture medium for lactic acid bacteria and fermenting them to obtain a fermented citrus fruit extract;
(B) obtaining an extract of citrus peel from citron;
(C) obtaining a gold extract from gold;
(D) obtaining an octopus fennel extract from octopus fennel; And
(E) mixing 1 to 60% by weight of the fermented citrus fruit juice extract, 1 to 30% by weight of citrus juice extract, 1 to 80% by weight of the golden extract and 1 to 60% by weight of the octopus fennel extract Gt; The method of claim 6,
The lactic acid bacteria may be selected from the group consisting of Lactobacillus brevis, Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus plantarum, Lactobacillus kimchii, Lactobacillus paraplantarum, Lactobacillus carbatus subcapsule. Lactobacillus curvatus subsp. Curvatus, and Lactobacillus subtilis. A method for the production of a natural preservative selected from one or more of Lactobacillus sakei subsp. Sakei.

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