WO2012008776A2

WO2012008776A2 - Method for preparing broussonetia kazinoki extracts, broussonetia kazinoki extracts prepared by the method, and cosmetic composition comprising the extracts

Info

Publication number: WO2012008776A2
Application number: PCT/KR2011/005191
Authority: WO
Inventors: 권순상; 이진영; 정한순; 김소연; 안성준
Original assignee: (주)아모레퍼시픽
Priority date: 2010-07-14
Filing date: 2011-07-14
Publication date: 2012-01-19
Also published as: WO2012008776A3; CN102985066B; CN102985066A

Abstract

The present invention relates to a method for preparing Broussonetia kazinoki extracts, to Broussonetia kazinoki extracts prepared by the method, and to a cosmetic composition comprising the extracts. More particularly, the present invention relates to a method for preparing Broussonetia kazinoki extracts, which involves extracting Broussonetia kazinoki using an ultra-high-pressure technique, a supercritical technique, or an oil-extraction technique, to thereby increase active ingredients and improve skin-whitening, anti-aging, anti-wrinkle, skin-moisturizing and skin-elasticity-enhancing effects. The present invention also relates to Broussonetia kazinoki extracts prepared by the method, and to a cosmetic composition comprising the extracts. In addition, the present invention relates to a method for fermenting Broussonetia kazinoki or extracts thereof, to fermented extracts prepared by the method, and to a cosmetic composition comprising the fermented extracts. More particularly, the present invention relates to a method for fermenting Broussonetia kazinoki or the extracts thereof, which involves naturally fermenting Broussonetia kazinoki or extracts thereof, or ferments Broussonetia kazinoki or extracts thereof by adding a fermentation strain or a predetermined amount of salt, and extracting the fermented Broussonetia kazinoki or extracts thereof using a solvent, to thereby prevent the propagation of various bacteria such as putrefactive bacteria, colon bacteria and anaerobic bacteria, which are harmful to the human body, remove offensive odors, and improve skin-whitening, anti-aging, anti-wrinkle, skin-moisturizing, and skin-elasticity-enhancing effects as compared to conventional extracts. The present invention also relates to fermented extracts prepared by the method, and to a cosmetic composition comprising the fermented extracts.

Description

【Specification】

[Name of invention]

Method for producing a mulberry extract, the mulberry extract produced by it and a cosmetic composition comprising the same

Technical Field

The present invention relates to a method for producing a mulberry extract, and to a cosmetic composition comprising the mulberry extract produced thereby. More specifically, the present invention relates to a method for producing extracts by increasing the active ingredient by extracting the mulberry or its extract by ultra high pressure, supercritical or oil extraction.

In addition, the present invention is fermented by adding natural fermentation strain or fermentation strain or a certain amount of salt, and then extract it with a solvent to prevent the growth of various bacteria such as decayed bacteria, Escherichia coli and anaerobic bacteria harmful to the human body, etc. It is about fermentation method to remove.

And thereby, whitening, anti-aging, anti-wrinkle, moisturizing and elasticity improvement effect than the existing extracts, a method of producing a mulberry extract, a mulberry extract and a cosmetic composition comprising the same, or a mulberry or fermentation method thereof It relates to a fermentation extract and a cosmetic composition comprising the same.

Background Art

The present invention relates to a method for producing a mulberry extract, a mulberry extract produced by it and a cosmetic composition comprising the same. More specifically, it relates to a method for producing extracts by increasing the active ingredient by extracting the mulberry or its extracts by ultra high pressure, supercritical or oil extraction method.

In addition, the present invention is fermented by adding natural fermentation or fermentation strains or a certain amount of salt, and then extracted with a solvent to prevent the growth of decayed bacteria, Escherichia coli and anaerobic bacteria, etc. harmful to the human body and odor, etc. It relates to a fermentation method to remove it.

And thereby, whitening, anti-aging, anti-wrinkle, moisturizing and elasticity improvement effect than the existing extracts, a method of preparing a mulberry extract, a mulberry extract and a cosmetic composition comprising the same, or a mulberry or fermentation method thereof It relates to a fermentation extract and a cosmetic composition comprising the same.

[Detailed Description of the Invention]

[Technical problem] The present inventors studied a method for increasing the content of the active ingredient in the mulberry extract, when the extract of the mulberry by ultra high pressure, supercritical or oil extraction method and the whitening, anti-aging, anti-wrinkle, moisturizing and elasticity improvement effect Discovered and completed the present invention. In addition, the fermented extracts of the mulberry or its extracts are naturally fermented or fermented with fermentation strains or a certain amount of salt, and then extracted with a solvent. Discovered and completed the present invention.

Accordingly, it is an object of the present invention to provide a method for preparing a mulberry extract for increasing the content of the active ingredient in the mulberry extract and to provide a cosmetic composition comprising a mulberry extract prepared by the above method.

It is also an object of the present invention to provide a fermentation method and its fermentation extract of the mulberry or its extract for increasing the content of the active ingredient in the mulberry extract.

Technical Solution

In order to achieve the above object, in the present invention, the preparation of the mulberry extract comprising the step of extracting at least one selected from the group consisting of root, stem, leaf, flower and fruit of the mulberry by ultra high pressure, supercritical or oil extraction method It provides a cosmetic composition comprising the method and the extract of the mulberry prepared by the above method.

In addition, the present invention provides a fermentation method for natural fermentation, homogeneous fermentation or salt fermentation of the mulberry or its extract and the fermented extract of the mulberry obtained by the above method.

[Favorable effect]

The mulberry extract extracted by the method according to the present invention was excellent in the whitening, anti-aging, anti-wrinkle, moisturizing and elasticity improvement effect due to the increase in the active ingredient.

In addition, the white fermented extract of fermented mulberry according to the present invention was also effective in improving whitening, anti-aging, anti-wrinkle, moisturizing and elasticity due to the increase of the active ingredient, and in the case of salted fermented extract, Escherichia coli or anaerobic bacteria It was possible to prevent the decay and contamination by the back.

[Brief Description of Drawings]

1 is a cross-sectional view of the ultra-high pressure used in the present invention.

Figure 2 is a photograph of the ultra-high pressure used in the present invention.

Figure 3 shows the effect of salt on the growth of salt in culture of Escherichia coli Escherichia coli ATCC 8739), Pseudomonas aeruginosa ATCC 9027), and Staphylococcus aureus ATCC 6538). Smear After confirming the inhibition ring generation results are shown.

[Best form for implementation of the invention]

The present invention relates to a method for preparing a mulberry extract by ultra high pressure, supercritical or oil extraction. Hereinafter, the present invention will be described in more detail.

The present invention relates to a method for producing a mulberry extract by ultra-high pressure, supercritical or oil extraction method, more specifically, the extraction method according to the present invention is water of at least one root, stem, leaf, flower and fruit of the mulberry Or extracting the active ingredient for 4 to 24 hours at 10 to 90 ton by adding a lower alcohol of C1 to C5; And extracting all the ultra-high pressure, supercritical or oil extraction method, the ultra-high pressure, supercritical or oil extraction method may be carried out simultaneously or sequentially with the extraction step, the extraction process using a lower alcoholol May be omitted.

In one embodiment of the present invention can be prepared by extracting the mulberry using ultra-high pressure technology. For example, the roots, stems, leaves, flowers, or fruits of the mulberry can be extracted by ultra-high pressure treatment for 10-20 minutes at a pressure of 50-300 Mpa at 20-50 ^° C, preferably 70 at 25-35 ^° C. Ultra high pressure treatment for 10 to 15 minutes at a pressure of 200 to 200 Mpa. In an exemplary embodiment of the present invention, an ultrahigh pressure process may be performed using an ultrahigh pressure device having the configuration shown in FIGS. 1 and 2, but is not limited thereto. In Figure 1, the ultra-high pressure of the cylinder (cyl inder), the processing chamber containing the pressurized material of the cylinder portion (Pressurizing Piston), the water purifying jacket (Adjusting the temperature outside the cylinder) In the lab scale, the maximum pressure of HHP is 600 Mpa and the working volume is about 0.6 ί. The possibility is a pilot lot-scale UHP plant (35 L) with a maximum pressure of 2,000 Mpa and a working volume of up to 35. In addition, in one embodiment of the present invention can be prepared by extracting the mulberry by using a supercritical technology. In this case, as the supercritical fluid, carbon dioxide, nitrogen, nitrous oxide, methane, ethylene, propane, and propylene may be used. Preferably, supercritical carbon dioxide is used. Supercritical fluidized carbon dioxide has a critical pressure of 73.8 bar and a low critical temperature of 31.11, making it easy to produce supercritical conditions in general. This is because carbon itself is not toxic and inexpensive.

The supercritical extraction conditions are preferably extracted by adding a supercritical fluid under a pressure of 20 ~ 40 ^° C, 70 ~ 200 bar. Extraction efficiency is increased when the extraction temperature is more than 20 ° C, extraction efficiency is higher than 40 C, but there is no increase in extraction efficiency compared to below. In addition, the extraction efficiency is increased when the air pressure is 70 bar or more, and there is no increase in the extraction efficiency when compared to the air pressure of less than 200 bar. In one embodiment of the present invention may be prepared by extracting the mulberry tree through the oil extraction method. For example, vegetable oils such as soybean oil and sesame oil may be added to the roots, stems, leaves, flowers or fruits of the mulberry tree and extracted at 40 to 70 ^° C and 2 to 10 bar pressure for 2 to L2 hours. In addition, the present invention relates to a cosmetic composition comprising a mulberry extract prepared by the above method in an amount of 0.0001 to 90% by weight, preferably 0.001 to 50% by weight based on the total weight of the composition. In addition, the cosmetic composition according to the invention is characterized in that for whitening, anti-aging, anti-wrinkle, moisturizing or elasticity improvement. On the other hand, the present invention relates to a method of fermenting the mulberry or its extract in order to increase the content of the active ingredient in the mulberry extract.

In the present invention, it may be fermented by a conventional method well known in the art. At this time, the mulberry extract can be prepared by extracting with water or ethanol by a method well known in the art. For example, water or ethanol may be added to the roots, stems, leaves, flowers or fruits of the mulberry tree and extracted for 10 to 90 ^° C for 4 to 24 hours. Preferably, water may be used. It is not limited. Hereinafter, the present invention will be described in more detail.

The present invention relates to a method for natural fermentation of mulberry or its extract. In one embodiment of the present invention, the mulberry or its extract can be prepared by mixing with sugar components in a container such as Onggi natural fermentation without artificial manipulation. At this time, the sugar component that can be used is not particularly limited, for example, sucrose, xyl, oligosaccharides, dietary fiber, erysri, glucose or fructose may be used. The sugar component is preferably mixed in 1 to 10% by weight based on the total weight of the fermented product, Fermentation is carried out for 30 days to 6 years, preferably 6 months or more in the natural environment of the four seasons. The present invention also relates to a method of homogenizing the mulberry or its extract. In one embodiment of the present invention, the mulberry or its extract is inoculated with useful fermentation strains well known in the art, such as yeast, mushroom, lactic acid, yeast or Bacillus genus (battery) 18 to 100 hours at 10 ~ 60 ^° C. It can be fermented for about a day. In this case, the fermented strains may be directly inoculated with Dakna radish and fermented, and then extracted with water or ethanol, or the fermented strains may be inoculated with the existing Dakna extract to obtain fermented products.

Yeast bacteria used in the present invention is Saccharomyces

sp.), Ski-jo！ "Horizontal τ ³ !" is-^) ― (Schi zosaccharomyces sp.), Tor u lops is sp.), Rhodotor ^^ Rhodotorula sp.) or Candida /? ( / ^ sp.), and mushroom fungi can be any edible fungi, preferably chaga mushroom (Inonnotus-obl iquus), neubacterial fungi (6 厂 sinensis, Paeci lomyces japonica) ),

For example, it can be used as a situation mushroom G¾? //// 7 / s linteus) or as a Gano / enna lucidum. In addition, lactic acid bacteria (Lactobaci 1 lus sp.), Streptococcus (5 rep ocOcci / s sp.), Leuconostoc sp.

sp.), and yeast bacteria are razopus usami (½ 7 ^' ) and florae, aspergillus duck

oryzae) or Aspergillus (? e / 7 / i / s sojae), and Bacillus genus (Bacillus subtilis) is Bacillus licheniformis

1 i chen i form is) F1017, Bacillus licheniformis lichen / form Is) F1232, Bacillus licheniformis 0¾c /// i / s 1 i chen i form is) F1267, Bacillus licheniformi ^ (Bacillus 1 i chen i form is) F1268, Bacillus licheniformis ¾ ^ 7 / ^ 1 i chen i form is) F1200, Bacillus pumilus pumilus) F1337, Bacillus sonorensis sonorensis) F1005, Bacillus subtilis CSscJ / i / s subtil is)

F1236 or Bacillus subtilis) F1279, and the like, but is not limited thereto.

When fermentation using yeast in one embodiment of the present invention, inoculation of yeast at a constant concentration (about 10 ⁵ to 10 ⁷ cell / mL) to the base medium containing the mulberry or its extract pH 5.5 ~ 6.5, temperature 25 2x while maintaining the condition of ~ 50 ^° C, aeration 0.05 0.5 VVM After acquiring, the fermentation broth can be obtained by centrifuging the fermentation broth to remove the polymer precipitate and the yeast. Incubation time is preferably 24 to 120 hours. In addition, when fermented with mushrooms in one embodiment of the present invention, after inoculating mushroom bacteria in the mulberry or its extract and incubated while maintaining the conditions of temperature 18-25 ^° C, humidity 60-80¾, water or carbon number 1-4 anhydrous or hydrous lower alcohols (e.g. methanol, ethanol, propanol and butanol), mixed solvents of the lower alcohols with water, acetone, ethyl acetate, chloroform, 1,3-butylene glycol, nucleic acid or One or more kinds of diethyl ether may be used, and then it is concentrated under reduced pressure by heating for 50 to lOO for 1 to 5 hours to obtain a fermented extract of Methanol. Incubation time is preferably 1 day to 100 days.

In addition, when fermented with lactic acid bacteria in one embodiment of the present invention, lactic acid bacteria inoculated at a constant concentration (about 10 ⁵ to 10 ⁷ cel l / mL) in the base medium containing the mulberry or its extract pH 6.5- After fermentation with 7.5, temperature 25-50 ^° C, and aeration of 1 WM, the fermentation broth can be centrifuged to obtain a fermented extract of Mt. Incubation time is preferably 24 to 120 hours.

In addition, when fermented with Nuruk in one embodiment of the present invention, inoculated with Nuruk bacteria in the mulberry or its extract and incubated while maintaining the temperature of 15 ~ 45 ΐ, centrifugation of the fermentation broth, polymer precipitate and Nuruk It can be obtained from the dried fermented extract of the mulberry. Incubation time is preferably 18 to 120 hours.

In addition, when fermented with Bacillus sp. In one embodiment of the present invention, Bacillus sp. Inoculated at a constant concentration (about 10 ⁵ to 10 ⁷ cel l / mL) to the base medium containing the mulberry or its extract After culturing while maintaining the temperature of 10 ~ 60 ° C, this fermentation broth can be obtained by centrifugation of the fermentation extract of the mulberry fermentation extract pH 5.5 ~ 8.5, polymer precipitates and Bacillus bacteria. Incubation time is preferably 24 to 96 hours. The present invention also relates to a method of fermenting salted mulberry or its extracts. In one embodiment of the present invention, the mulberry or its extract is 1) adding a certain amount of salt to the mulberry or its extract for the long-term aging fermentation; And 2) extracting the salted fermented product with water or an organic solvent to obtain a fermentation extractol.

In the step 1), as the salt used for salting, refined high purity sodium chloride, sun salt, rock salt, bamboo salt, and the like are preferable, and the salt concentration is preferably 10-30 increase ¾ with respect to the whole fermented product. When salt content is less than 10% by weight, we expect It is difficult to do, and when the amount exceeds 30% by weight, the effect is not changed.

The fermentation process of the present invention is preferably fermented 30 days to 1 year, preferably 6 months to 1 year so that sufficient fermentation can occur between 4 to 40 ^° C. If it is less than 4 ^° C, growth of useful microorganisms is poor, and if it exceeds 40 ° C, the growth of microorganisms may be excessive, affecting the performance of the mulberry or its extract.

The extraction solvent used in step 2) may be selected from the group consisting of purified water, methanol, ethanol, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane and nucleic acid alone or mixed two or more kinds. Can be used. At this time, the extraction temperature is preferably 10 to 80 ^° C, it can be extracted for 6 hours to 24 hours. If the extraction temperature and the extraction time is out of the extraction efficiency may decrease or change of components may occur.

In the present invention, after the step 2) in a conventional method known in the art can be prepared by salting fermented extract of the mulberry by evaporation, spray drying or lyophilization of the liquid obtained by cooling, heating and filtration in phase silver or additional solvent. .

In another aspect, the present invention relates to a cosmetic composition comprising 0.0001 to 90% by weight, preferably 0.001 to 50% by weight, based on the total amount of the mulberry fermented extractol composition prepared by the above method. In addition, the cosmetic composition according to the invention is characterized in that for whitening, anti-aging, anti-wrinkle, moisturizing or elasticity improvement.

[Form for implementation of invention]

Hereinafter, the content of the present invention will be described in more detail through Examples and Test Examples. These examples are provided only for understanding the contents of the present invention, but the scope of the present invention is not limited to these examples, and modifications, substitutions, and insertions that are commonly known in the art may be performed. It is also included in the scope of the present invention.

Example 1 Preparation of Ultra High Pressure Extract

After washing 1 kg of the stem and roots of the mulberry tree with clean water and drying it, prepare the ultra-high pressure treatment by setting the ultra-high pressure machine, fill it with water in the processing chamber, and pack the mulberry prepared above in a vacuum pack. After treatment at a temperature of 501C or less for 1 minute or more, cooled to room temperature and dried in a dryer to obtain a extract of the mulberry. Example 2 Supercritical Extract Preparation

After washing 1 kg of the stems and roots with dry water, drying them, and putting them in a supercritical extraction tank, supplying carbon dioxide to the extraction tank using a high-pressure gas pump stopped supplying carbon dioxide when the pressure reached 75 bar. The temperature inside the extraction tank was heated to 40 ^° C. at a rate of 2 / min through a heating wire surrounding the outside of the extraction tank using a temperature controller. Using a pressure regulator attached to the outlet of the extraction tank, the carbon dioxide was partially discharged and maintained at 34 ^° C. and 74 bar for 30 minutes. Then, the high pressure gas pump was restarted to continuously supply carbon dioxide to the extraction tank at a flow rate of 2 L / min. At the same time, the extract was released using a pressure regulator attached to the outlet side, and the temperature of the extraction tank and the pressure were kept constant at 34 ^° C 74 bar. While maintaining the temperature of the pressure regulator at 40 ^° C, the emissions released through the pressure regulator transferred carbon dioxide to solid dry ice due to pressure and temperature drop. The released extract was decompressed to atmospheric pressure to release carbon dioxide into the atmosphere, and the mulberry extract was recovered into the collector. The above procedure was carried out for 4 hours to obtain a mulberry extract.

Example 3 Oil Extract Preparation

1 kg of gingko stems and roots were washed with clean water, dried and soaked in 10 grams of soybean oil, and extracted at 50 ^° C for 2 hours at a pressure of 2 to 4 bar in an extractor equipped with a cooling condenser and filtered through a filter paper. The extract was concentrated under reduced pressure at 70 ° C. in a distillation apparatus equipped with an angle capacitor to obtain a extract of the mulberry oil.

Comparative Example 1

After washing 1 kg of gingko stem and root with dry water, dried it, put it in water 10, boil it for 5 hours in an extractor with cooling capacitor, extract it, filter it with 300 mesh filter cloth, and leave it for 5 days at 15 ° C for 5 days to mature. Filter with Whatman twice filter paper. The extract was concentrated under reduced pressure to 8 (C under reduced pressure in a distillation apparatus equipped with a cooling capacitor to obtain 71.3 g (dry weight).

Example 4

The mulberry extract prepared in Comparative Example 1 was subjected to ultra high pressure in the same manner as in Example 1. Example 5

Supercritical extraction of the mulberry extract prepared in Comparative Example 1 in the same manner as in Example 2.

Example 6

The mulberry extract prepared in Comparative Example 1 was extracted with oil in the same manner as in Example 3.

Test Example 1 Tyrosinase Inhibitory Effect

Tyrosinase enzyme was extracted from the mushroom (Mushroom) was used as that of Sigma (SIGMA). First, the substrate tyrosine was dissolved in distilled water to make a 0.3 mg / ml solution, and 1.0 ml of the solution was added to the test tube, followed by 1.0 ml of potassium-phosphate buffer solution (0.1 mol concentration, pH 6.8) and distilled water. 0.7 ml was added.

0.2 ml of each extract sample solution prepared by mixing the extracts of Examples 1 to 6 and Comparative Example 1 in an appropriate concentration in an ethanol solution and then reacted for 10 minutes in a 37 ^° C thermostat. At this time, the control group was prepared by adding only 0.2 ml of solvent instead of each extract, and ascorbic acid was used as a positive control. 0.1 ml of 2500 units / ml tyrosinase solution was added to the reaction solution, and the reaction mixture was reacted for 10 minutes in a 37C thermostat. The test tube containing the reaction solution was quenched by immersing it in iced water, and the absorbance at wavelength 475 nm was measured using a photospectrometer. The results are shown in Table 1 below. Tyrosinase inhibition effect of each extract was calculated by the following equation (1).

[Equation 1]

EJ Rossinase IHS (% I = 10

Table 1

In the results of Table 1, it was found that the high tyrosinase inhibition rate of the mulberry extract of Examples 1 to 6 prepared by ultra high pressure, supercritical and oil extraction method is excellent in the whitening effect.

Test Example 2 Evaluation of Melanin Inhibitory Activity Using B16 / F10 Melanoma Cells

Examples 1 to 6, Comparative Example 1 and a sample containing 0.001% by weight of kojic acid, respectively, were added as a test substance to the culture medium of B16 / F10 melanoma cells (Korea Cell Line Bank) at a constant concentration and then cultured for 3 days. The culture solution was removed, washed with PBS and the cells were dissolved in IN NaOH and absorbance was measured at 405 nm. As a control, the cells without addition of the test substance were compared with the melanin content in the control group, and the degree of inhibition of melanin production of each test substance was measured. According to Equation 2, the melanin production inhibition rate was calculated and the results are shown in Table 2.

[Equation 2]

Table 2

From the results of Table 2, ultra-high pressure, supercritical, and a yarn prepared by the oil extraction入^'Example 1 Melanogenesis inhibition rate of 6 mulberry extract of a high whitening effect is seen Great丁·

Example 7 Preparation of Natural Fermented Extract of Methanol

50 kg of oligosaccharides were washed with 1 kg of dried oak stems and roots, and when they were filled, they were pressed without pressure by pressing them from the top, and naturally fermented for 6 months. After the fermentation is finished, the fermented raw material is filtered through S ^ By removing the dregs, the natural fermented extract of the mulberry was obtained.

Example 8 Fermented Extract of Methanol Using Yeast

By adding soybean extract powder of Comparative Example 1 to 5% by weight in 1.5 purified water, the basic composition of carbon source, nitrogen source, ginseng salt, inorganic salts and micronutrient components, and suitable pH 5.5 ~ 6.5 using sodium hydroxide Set to. Saccharomyces (5 ^ c? Arozzyces sp.) Is inoculated with ⁶ cell / mL to the culture medium containing the mulberry extract powder, pH 5.5 ~ 6.5, temperature 25 ~ 35 ^° C, aeration amount 0.5 WM Incubated for 72 hours while maintaining the conditions of.

The fermentation broth was centrifuged at l.OOOXg for 30 minutes to obtain a final fermented extract of all isolates from which polymer precipitates and yeast cells were removed.

[Example 9] Fermented Extract of Methanol Using Mushroom Bacteria

To the mulberry extract powder of Comparative Example 1 was added 5 wt ¾ of the substratum fungus (/ ¾ec / 7c «? Yces japonica), and fermented at 18-25 ^, humidity 60-70¾ for 60 days, dried and then 100 g of the pulverized product obtained by grinding was put into 80% methanol 3, respectively, and boiled for 3 hours in a reflux extractor with a cooling condenser. Then filtered through 100 mesh filter cloth and the residue was extracted once more in the same way. Each extract was combined and filtered at room temperature with Whatman No. 2 filter paper to remove insoluble materials, concentrated under reduced pressure at 60 ^° C in a distillation apparatus equipped with a cooling condenser, and lyophilized to obtain the final mulberry fermentation extract.

[Example 10] Fermented Extract of Methanol Using Lactic Acid Bacteria

By adding soybean extract powder of Comparative Example 1 to 5% by weight in 1.5 purified water, the carbon composition, nitrogen source, ginseng salt, inorganic salts, micronutrient components, etc. as a basic composition, using a sodium hydroxide with an appropriate pH 5.5 6.5 Set to. Lactobacillus in the culture medium containing the mulberry extract powder

s / was inoculated at 10 ⁶ cell / mL, and cultured for 72 hours while maintaining the conditions of pH 6.5-7.5, temperature 35-38t, aeration rate 1 WM.

The fermentation broth was obtained by centrifugation for 30 minutes in l.OOOXg to remove the final mulberry fermentation extractol from which polymer precipitates and lactic acid bacteria cells were removed. [Example 11] Fermented Extract of Korean Dillwood Using Nuruk

Aspergillus duckwood isper ᅳ oryz e) was added to 5% by weight of the extract powder of Comparative Example 1, fermented at 25 ^° C. for 48 hours, dried, and then ground. Each was put in 80% methanol 3 and extracted by boiling in a reflux extractor with a cooling capacitor for 3 hours. Then filtered with 100 mesh filter cloth and the residue was extracted once more in the same way. Each extract was combined and filtered with Whatman No. 2 filter paper at room temperature to remove insoluble matters, and then concentrated under reduced pressure in a distillation apparatus equipped with a cooling condenser and lyophilized to obtain a final mulberry fermentation extract.

[Example 12] Fermented Extract of Methanol Using Bacillus Bacillus

By adding soybean extract powder of Comparative Example 1 to 5% by weight in 1.5 purified water, the basic composition of carbon source, nitrogen source, ginseng salt, inorganic salts and trace nutrients, etc., and using appropriate pH 5.5-6.5 Set to. Bacillus subtilis 0 c / / / s subtil is) F1279 was inoculated in the culture medium containing the mulberry extract powder, and incubated for 72 hours at 45 C.

The fermentation broth was centrifuged at LOOOXg for 30 minutes to obtain a final mulberry fermentation extract from which polymer precipitates and Bacillus cells were removed.

Example 13 Preparation of Methanol Salt Fermented Extract

1 kg of gingko stems and roots, washed with clean water and dried, mixed in 10 weight «> concentration of salt solution, prepared in pots, fermented at 4 ° C for about 30 days, and then added 5 £ of 80% aqueous ethanol solution ， 3 times reflux extraction, and then 1 day at 15 ^° C. Thereafter, the residue and the filtrate were separated by filtration and centrifugation of the filter cloth. The separated filtrate was suspended in an extractol obtained by concentrating under reduced pressure, extracted five times with ether 1 to remove the pigment, and the aqueous layer was 1-butanol. Extracted three times with 500 m «. The resulting 1-butanol extract was concentrated under reduced pressure to obtain 1-butanol extract, which was dissolved in a small amount of methanol, and added to a large amount of ethyl acetate. The resulting precipitate was dried and 50.3 g of salted fermented extract of Methanol was obtained. It was.

Test Example 3 Tyrosinase Inhibitory Effect

Tyrosinase enzyme was extracted from the mushroom (Mushroom) was used by SIGMA (SIGMA). First, tyrosine, a substrate, was dissolved in distilled water to prepare a 0.3 mg / ml solution. 1.0 ml of the solution was added to the test tube, and then 1.0 ml of potassium-phosphate buffer solution (0.1 mol concentration, pH 6.8) and 0.7 ml of distilled water were added thereto.

0.2 ml of each extract sample solution prepared by mixing the extractol ethanol solution of Examples 7-13 and Comparative Example 1 in an appropriate concentration was added to the reaction solution and reacted for 10 minutes in a 37 ^° C thermostat. At this time, the control group was prepared with 0.2 ml of solvent instead of each extract, and ascorbic acid was used as a positive control. 0.1 ml of a Tyrosinase solution of 2500 units / ml in this reaction solution was added and reacted again in a 37 ^° C thermostat for 10 minutes. The reaction tube containing the reaction solution was placed in iced water and rapidly dropped to stop the reaction. The absorbance at the wavelength of 475 nm was measured using a photospectrometer. The results are shown in Table 3 below. The tyrosinase inhibitory effect of each extract was calculated by the above formula (1).

[Table 3]

In the results of Table 3, it was found that the tyrosinase inhibition rate of Examples 7 to 13, which is the Methanol fermentation extract according to the present invention, was high, and the whitening effect was excellent.

Test Example 4 Evaluation of Melanin Inhibitory Activity Using B16 / F10 Melanoma Cells

Example 7-13, Comparative Example 1 and a sample containing 0.001% by weight of kojic acid, respectively, were added as a test substance to the culture medium of B16 / F10 melanoma cells (Korea Cell Line Bank) at a constant concentration and then cultured for 3 days. The culture solution was removed, washed with PBS and the cells were dissolved in IN NaOH and absorbance was measured at 405 nm. As a control, the cells without addition of the test substance were compared with the melanin content of the control group, and the melanin production inhibition of each test substance was measured. The melanin production inhibition rate was calculated according to Equation 2 and the results are shown in Table 4.

Table 4

In the results of Table 4, the high whitening effect of melanin production of Examples 7-13, which is the Methanol fermentation extract according to the present invention was found to be excellent.

[Test Example 5] Harmful bacteria production inhibition experiment

E. coli (Escherichia coli ATCC 8739), Pseudomonas aeruginosa ATCC 9027), and yellow grapes (Staphylococcus aureus ATCC 6538) were pre-cultivated after first pouring and solidifying the medium (Letheen Agar) into the sale. The test bacteria to 40

^The medium adjusted to about ^° C (Letheen Agar) was finally sintered so that the amount of bacteria was 1 () ⁵ / g and poured on the primary medium. A sterilized paper disk (paper disk) 10%, 20% and 30% of the solar salt solution come 0.02 ml injection to be put on the bacteria plated medium solution is sufficiently diffused at room temperature for 24 hours and then at each 32 ^° C After culturing for an optimal time, it was confirmed whether the inhibitory ring formed in the saale was produced, and the results are shown in FIG. 3.

From the results of Figure 3, it was confirmed that the salt inhibits the growth of harmful microorganisms.

[Formulation Example 1 13 and Comparative Formulation Examples 1 and 2]

Nutritional creams of Formulation Examples 1 to 3 and Comparative Examples 1 and 2 were prepared in a conventional manner according to the compositions of Tables 5 to 7 below (unit: weight%).

Table 5

[Table 6]

[Table 7]

Exfoliating ingredient comparator Ί comparator ¾ example 2 comparative example 1 0.5 ¬ lead 10 10

Polysomate 60 1.5 1.5 Fiji 60 Cured Castor Oil 2 2

Sombitases du sound rate 0.5 0.5 Huedu paravi 10 10

Scuamla 5 5

Caprylic / capric triglyceride (Estasan: Uniqema (5 5

manufacturer))

Glycerol 5 5

Wealth Recall 3 3

Pro ¾ Freedom Recall 3 3

Triethanol? 1 0.2 0.2 preservative. ^ Bovine and flavor appropriate amount purified water [Test Example 6] The skin moisturizing effect of the extract of the mulberry prepared by the ultrahigh pressure, supercritical and oil extraction method according to the present invention

In order to measure the effect of the extract of the mulberry extract according to the present invention on the skin moisturizing, 160 adult men and women in their 40s to 50s showing skin dryness divided into eight sets Formulation Examples 1-6 and Comparative Formulation Examples 1-2 Was applied to the face twice daily for four weeks. Coniometer at constant temperature, constant humidity (24 ^° C, 40% relative humidity) before start of application, after 1 week, 2 weeks, 4 weeks after application, and after 2 weeks (total 6 weeks) The skin moisture content was measured, and the results are shown in Table 8 below. The conniometer is a skin moisture meter that measures the amount of moisture present on the skin by measuring the conductivity of the epidermis. The test result is a percentage of the increase of the measured value after a certain period of treatment based on the Koniometer value measured immediately before the test was started.

[Table 8]

In the results of Table 8, it was confirmed that the skin moisture increase rate of the formulations 1 to 6 containing the mulberry extract of Examples 1 to 6 prepared through the ultra high pressure, supercritical and oil extraction method according to the present invention. . In addition, even after stopping the application of the nutrition creams of Formulation Examples 1-6, the measured value of skin moisture was similar to that of 1 week after application, and it was found that the skin moisture was maintained for a certain period without applying the Example. .

Test Example 7 Skin Wrinkle Improvement Effect of Methanol Extract Prepared by Ultra High Pressure, Supercritical and Oil Extraction Method of the Present Invention

In order to measure the effect of the mulberry extract according to the present invention on the skin wrinkle improvement, the effect of improving skin wrinkles was compared to 160 subjects with facial wrinkles aged 30 to 50 years. Measure the skin condition on both sides of the face before using the cream, and then instruct each subject to use the creams of Formulation Examples 1-6 and Comparative Formulations 1-2 for 3 months, and then re-measure the same area for skin wrinkles. The amount of change was measured, and the results are shown in Table 9 below. The skin wrinkles were measured with a Visiometer system (C + K) in a constant temperature and humidity room of 24 ^° C. and a relative humidity of 40%. The amount of change in skin wrinkles was calculated according to the following equation.

[Equation 3]

In Equation 3, "Tdi" is a measurement site value at D90, and "Tdo '' is a measurement site value at DO.

Table 9

In the results of Table 9, it was confirmed that the skin wrinkle improvement effect of Formulation Examples 1 to 6 including the mulberry extract of Examples 1 to 6 prepared through the ultrahigh pressure, supercritical and oil extraction method according to the present invention. .

Test Example 8 Confirmation of Skin Elasticity Improvement Effect of Methanol Extract Prepared by Ultra High Pressure, Supercritical and Oil Extraction Method According to the Present Invention

In order to confirm the effect of improving skin elasticity, the creams of Formulation Examples 1 to 6 and Comparative Formulation Examples 1 to 2 were measured by dividing the cream into 8 groups of 20 people in 160 women in their 30s to 40s. The experimental method is as follows.

Disperse each cream of Formulation Examples 1 to 6 and Comparative Formulation Examples 1 to 2 and apply it to the face once a day for 12 weeks, and then use a skin elasticity measuring instrument (Cutometer SEM 575, C + K Electronic Co., Germany). Skin elasticity was measured using. The results are shown in Table 10 below. The results in Table 10 show the AR8 (R8 (left) -R8 (right) of the Cutometer SEM 575. Value), the R8 value indicates the nature of the skin viscoelasticity.

Table 10

Looking at the results of Table 10, it was confirmed that the skin elasticity improving effect of the formulations 1 to 6 containing the mulberry extract of Examples 1 to 6 prepared through the extraction process according to the present invention.

Therefore, it can be confirmed that the cosmetic composition containing the mulberry extract of the present invention is very effective in improving the skin elasticity.

[Test Example 9] The skin moisturizing effect of the extract of fermented young mulberry according to the present invention

In order to measure the effect of the fermented extract of Dakberry according to the present invention on the increase of skin moisturizing, 180 groups of adult men and women in their 40s to 50s showing skin dryness were divided into 9 groups of Formulation Examples 7 to: L3 and Comparative Formulation Examples 1 to 2 Was applied twice a day for 4 weeks on the face. Coniometer at constant temperature, constant humidity (24 ^° C, 40% relative humidity) before start of application, after 1 week, 2 weeks, 4 weeks after application and after 2 weeks (total 6 weeks) after application was stopped. The skin moisture content was measured, and the results are shown in Table 11 below. The Koniometer is a skin moisture meter that measures the amount of electrical conductivity of the epidermis and the amount of moisture present in the skin. The test result is a percentage of the increase in the measured value after a period of treatment based on the Koniometer value measured immediately before the test commencement.

Table 11

Test Material Moisture Growth Rate (%)

1 week 2 weeks 4 weeks 6 weeks Formulation Example 7 33 41 42 31

Formulation Example 8 32 40 42 30

Formulation Example 9 33 39 40 29

Formulation Example 10 30 36 38 30

Formulation Example 11 31 37 37 32

Example 12 31 37 40 31

Formulation Example 13 32 38 38 29

Comparative Formulation Example 1 30 32 32 15

Comparative Formulation Example 2 32 35 38 26

In the results of Table 11, the embodiment prepared through the fermentation process according to the present invention

It was confirmed that the skin moisture increase rate of the formulation examples 7 to 13 containing the 7-13 of the mulberry fermentation extract. In addition, even after the application of the nutrition cream of Formulation Examples 7-13, the measured value of skin moisture was similar to that of 1 week after application, and it was found that the skin moisture was maintained for a certain period without applying the Example. .

Test Example 10 Skin Wrinkle Improvement Effect of Methanol Fermented Extract According to the Present Invention

In order to measure the effect of the fermented extract of Dakberry according to the present invention on skin wrinkle improvement, the skin wrinkle improvement effect was compared to 90 subjects with facial wrinkles aged 30 to 50 years.

Measure the skin condition on both sides of the face before using the cream, and then instruct each subject to use the creamols of Formulation Examples 7-13 and Comparative Formulations 1-2 for 3 months, and then re-measure the same area for skin wrinkles. The amount of change was measured and the results are shown in Table 12 below. In the constant temperature and humidity room of 24 ^° C and 40% relative humidity, the wrinkles of the tail of the eyes were floated with replicas and the skin wrinkles were measured by Visiometer system (C + K). The amount of change in skin wrinkles was calculated according to Equation 3 above.

Table 12

丄

In the results of Table 12, it was confirmed that the skin wrinkle improvement effect of Formulation Examples 7-13 including the mulberry fermented extract of Examples 7-13 manufactured through the fermentation process according to the present invention.

Test Example 11 Confirmation of Skin Elasticity Improvement Effect

In order to confirm the effect of improving skin elasticity, the creams of Formulation Examples 7 to 13 and Comparative Formulation Examples 1 and 2 were measured by dividing the cream into 9 groups of 20 people in a group of 180 women in their 30s to 40s. The experimental method is as follows.

Disperse each cream of Formulation Examples 7-13 and Comparative Formulation Examples 1 and 2 into each bath and apply it to the face once a day for 12 weeks, using a skin elasticity measuring instrument (Cutometer SEM 575, C + Electronic Co., Germany). Skin elasticity was measured. The results are shown in Table 13 below. The results in Table 13 are described as AR8 (R8 (left) -R8 (right)) values of Cutometer SEM 575, where R8 values indicate the nature of skin viscoelasticity.

Table 13

Looking at the results of Table 13, it was confirmed that the skin elasticity improving effect of the formulation examples 7 to 13 containing the mulberry fermented extract of Examples 7 to 13 prepared through the fermentation process according to the present invention.

Therefore, it can be confirmed that the cosmetic composition containing the fermented extract of the mulberry of the present invention is very effective for improving skin elasticity.

Claims

[Range of request]

[Claim 1]

A method for producing a mulberry extract, comprising the step of extracting at least one selected from the group consisting of root, stem, leaf, flower and fruit of a mulberry by ultra high pressure, supercritical or oil extraction.

[Claim 2]

The method of claim 1,

Method of producing a mulberry extract further comprising the step of extracting the active ingredient using water or C1 to C5 lower alcohols selected from the group consisting of the root, stem, leaf, flower and fruit of the mulberry ■

[Claim 3]

The method of claim 2,

The extraction process using the lower alcohol is a high-pressure, supercritical or oil extraction method that proceeds simultaneously or sequentially with the extract of the mulberry.

[Claim 4]

The method of claim 1,

The ultra-high pressure extraction method is a method of producing a mulberry extract that is treated for 1 to 15 minutes at a pressure of 70 to 200 Mpa.

[Claim 5]

The method of claim 1,

The supercritical extraction method is a method of producing a mulberry extract that is treated under a pressure of 20 ~ 40 ^° C, 70 ~ 200 bar.

[Claim 6]

The method of claim 5,

The supercritical fluid is carbon dioxide, nitrogen, nitrous oxide, methane, ethylene, propane or propylene manufacturing method of the mulberry extract.

[Claim 7]

According to that clause 1

The method of extracting oil is a method of producing a mulberry extract that is treated for 2 to 12 hours at 40 ~ 70 ^° C, 2 ~ 10 pressure by adding vegetable oil.

[Claim 8]

The method of claim 7, The vegetable oil is a soybean oil or sesame oil is a manufacturing method of the mulberry extract.

[Claim 9]

A cosmetic composition comprising a mulberry extract prepared according to any one of claims 1 to 8.

[Claim 10]

The method of claim 9,

The composition is a cosmetic composition for whitening, anti-aging, anti-wrinkle, moisturizing or improving elasticity

[Claim 11]

Fermentation method for natural fermentation, homogeneous fermentation or salt fermentation of mulberry or its extract.

[Claim 12]

According to claim 11,

The natural fermentation is a fermentation method of fermenting the mulberry or its extract with sugar components.

[Claim 13]

The method of claim 11,

The sugar component is at least one selected from the group consisting of sucrose, xyl, oligosaccharide, dietary fiber, erythritol, glucose and fructose.

[Claim 14]

The method of claim 11,

The fermentation method is a fermentation method of inoculating fermentation strains on the mulberry or its extracts and fermentation at 10 ~ 60 ^° C for 18 hours to 100 days.

[Claim 15]

The method of claim 14,

The fermentation strain is a fermentation method selected from the group consisting of yeasts, mushrooms, lactic acid bacteria, yeast bacteria and Bacillus genus bacteria.

[Claim 16]

The method of claim 11, wherein the salt fermentation

1) adding fermented salt to the mulberry or its extract for long-term fermentation;

2) extracting the salted fermented product with water or an organic solvent to obtain a fermented extract; Fermentation method which will include

[Claim 17]

The method of claim 16,

The salt used in step 1) is a fermentation method selected from the group consisting of high purity sodium chloride, sun salt, rock salt and bamboo salt.

[Claim 18]

The method of claim 16,

In the step 1), the amount of salt used is a fermentation method of 10-30% by weight relative to the total amount of fermentation.

[Claim 19]

The method of claim 16,

The organic solvent used in step 2) is at least one selected from the group consisting of methanol, ethanol, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane and nucleic acid.

[Claim 20]

The method of claim 16,

The fermentation process of step 1) is a fermentation method that is fermented from 4 to 40: 30 days to 1 year.

[Claim 21]

Fermented extract of Methanol prepared by the fermentation method according to any one of claims 11 to 20.

[Claim 22]

Cosmetic composition comprising the extract of fermented turmeric according to claim 21.

[Claim 23]

The method of claim 22,

The composition is a cosmetic composition for whitening, anti-aging, anti-wrinkle, moisturizing or elasticity improvement