KR101980684B1 - Method For Producing Ginseng Flower Extract With Enhanced Ginsenoside F4 Content and The Cosmetic Composition Including The Sam - Google Patents

Abstract

The present invention relates to a method for producing a ginseng flower extract having an increased content of ginsenoside F4, comprising a step of treating the ginseng flower with an enzyme followed by drying, a step of steaming the dried ginseng flower, and a step of extracting the steamed ginseng. The ginseng flower extract produced in the present invention is excellent in skin antioxidant and wrinkle-reducing effects by converting ginsenoside, which is low in bioavailability contained in ginseng, to ginsenoside F4 having high bioavailability.

Description

      FIELD OF THE INVENTION [0001] The present invention relates to a method for producing ginseng flower extract having enhanced ginsenoside F4 content and a cosmetic composition containing the ginsenoside F4 content and the cosmetic composition containing the ginseng flower extract.

The present invention relates to a method for producing ginseng extract having enhanced content of ginsenoside F4.

Ginseng ( Panax ginseng CA Meyer ) is a herb that belongs to the genus Ogawa and Ginseng. It has been used in Korea, China and Japan for over 2,000 years and has been used to prevent disease and prolong its life. The effects of ginseng on the central nervous system, anticancer activity, anti-cancer activity, immune function regulation, anti-diabetic effect, hepatic function hyperactivity, cardiovascular disorder improvement, anti-arteriosclerosis effect, blood pressure control effect, , Effects on osteoporosis, anti-stress action, anti-fatigue action, antioxidant activity, and inhibition of aging.

Ginseng contains various components useful for human body such as polyacetylene, phenolic compounds, acid polysaccharides, peptides, and alkaloids, which are non-saponin-based physiologically active substances including more than 40 kinds of glycosides such as ginsenosides.

Ginsenoside, a representative physiologically active ingredient of ginseng, refers to saponin contained in ginseng, and ginseng contains various kinds of ginsenosides. Ginsenosides are distributed evenly on the ground and underground of ginseng. Especially, it is known that not only ginsenoside content but also composition is different depending on sites such as ginseng roots, ginseng leaves and ginseng fruit.

The ginsenosides of ginseng constitute about 5% of the ginseng dry matter. The Ginsenosides of the ginseng contain about 5% of the ginseng in the Triterpenoid series of Dammarane skeleton containing glucose (Glucose), Arabinose, Xylose, (Rhamnose) and the like. (3, 6, 12 positions) Protopanaxadiol (PPD) saponins, 3 individuals (positions 3, 6 and 12) in the case of two individuals (positions 3 and 12) (Protopanaxatriol; PPT) family saponins. Glucose, Arabinose, Xylose, Rhamnose and the like are bonded to -OH in the R1, R2 and R3 positions of PPD and PPT. More than 30 kinds of ginsenosides have been reported to be isolated from ginseng. However, when extracting and analyzing actual ginseng, 6 kinds of Rb1, Rb2, Rc, Rd, Re and Rg1 account for more than 90% The content of saponin components is low.

Ginsenosides are not absorbed directly in the human body but are degraded by microorganisms such as Bacteriodes, Lactobacillus, Fusobacterium, Eubacterium, and Provetella species present in the human body and the metabolites are absorbed. In the case of ginsenoside Rb1, it is sequentially converted to ginsenoside Rd, compound K (hereinafter abbreviated as CK) and protopanaxadiol by intestinal microorganisms. Rb1, Rb2, Rc, and Rd, which are PPD-derived ginsenosides, are metabolized and absorbed by 20-O- β-D-glucopyranosyl-20 (S) -protopanaxadiol (CK) in the intestine. PPT ginsenosides Re, Rf, and Rg1 are metabolized in the intestine to Rh1, F1, and Protopanaxatriol. The PPD family ginsenosides Rb1, Rb2, Rc and Rd are converted into 20 (R) - / 20 (S) -ginsenoside Rg3 by intestinal microorganisms and Rg3 is converted into Rh2 by intestinal microorganisms.

In recent years, the effective ingredient of ginseng is easily absorbed into the human body, and a method of fermenting ginseng using intestinal microorganism or lactic acid bacteria or treating an enzyme with a trace amount of ingredients in ginseng is used. Korean Patent Publication No. 10-1017378 discloses a method for producing fermented red ginseng and fermented red ginseng and a Bacillus subtilis strain derived from chungkukjang, which is suitable for the fermented ginseng and fermented red ginseng, using a microorganism belonging to the genus Bacillus isolated from chungkukjang. And fermenting red ginseng to increase the content of ginsenosides Rd. The present invention also relates to a method for producing fermented ginseng and fermented red ginseng, and a technology for producing Bacillus subtilis strain derived from chungkukjang.

Korean Patent Registration No. 10-1240192 discloses a fermented ginseng extract for improving insulin secretion using Lactobacillus acidophilus strain K-59 and strain K-59 and a method for producing the same. Rg1, Rb1, Rg1 or Re, which is a low bioavailability component of ginseng, is converted into ginsenosides Rh1, Rg3, Rh2 or CK ( Lactobacillus acidophilus ) by using Lactobacillus acidophilus K-59 (KFCC11467P) A method for converting the above-mentioned compound is disclosed.

Accordingly, the present inventors have found that when extracts are prepared by enzymatic treatment and steam treatment using ginseng flower, ginsenoside having low bioavailability contained in ginseng is used as ginsenoside F4 having high bioavailability And the skin antioxidant and wrinkle-improving effect can be imparted. Thus, the present invention has been completed.

Korean Patent Publication No. 10-1017378 Korean Patent Publication No. 10-1240192

The main object of the present invention is to provide a method for converting ginsenoside having low bioavailability contained in ginseng flower into ginsenoside having high bioavailability.

In order to accomplish the above object, the present invention provides a method for manufacturing a ginseng flower, comprising the steps of: The enzyme treatment and the steaming to increase ginsenoside F4; And extracting the steamed ginseng flower. The present invention also provides a method for producing ginseng extract having enhanced content of ginsenoside F4.

In the present invention, the ginseng flower is characterized in that the ginseng flower is mixed with a side bottle.

In the present invention, the enzyme is characterized by being a pectinase and a cellulase.

In the present invention, the steaming treatment is characterized by treating steam.

In the present invention, the extracting step is performed by extracting expanded ginseng with ethanol (EtOH) to prepare an extract, followed by fractionation using HP20 porous resin and ethanol (EtOH).

The present invention also provides a ginseng extract prepared through the above-described method.

In the present invention, the ginseng extract has an effect on improving wrinkles.

The present invention also provides a cosmetic composition comprising the ginseng flower extract.

The ginseng flower extract prepared from the production method of the present invention is excellent in antioxidant and wrinkle-reducing effects because ginsenoside contained in ginseng is converted into ginsenoside F4 having high bioavailability.

FIG. 1 is a graph showing the Minor ginsenoside content of the ginseng extract obtained from Example 1 and Comparative Example 1 of the present invention. FIG.
2 is a graph showing cell viability results of the ginseng extract obtained from Example 1 and Comparative Example 1 of the present invention.
3 is a graph showing the DPPH (2,2-Diphenyl-1-picrylhydrazyl) scavenging activity of the ginseng extract obtained from Example 1 and Comparative Example 1 of the present invention.
4 is a graph showing the MMP-1 (Matrix Metallopeptidase-1) inhibitory activity of the ginseng extract obtained from Example 1 and Comparative Example 1. Fig.

Accordingly, when the ginseng flower is used and the ginseng flower extract is prepared by enzymatic treatment and steam treatment thereof, ginsenoside having low bioavailability contained in ginseng is converted into ginsenoside F4 having high bioavailability .

In this example, the ginseng was sequentially subjected to enzyme and bulging treatment using flowers, and then ginseng flower extract was prepared to confirm the increase of F4 of the specific ginsenoside.

Accordingly, in one aspect, the present invention provides a method for treating ginseng flower, comprising: enzymatically treating and drying ginseng flower; Boiling the dried ginseng; And extracting the ginseng grown. The present invention also relates to a method for producing ginseng flower extract having enhanced content of ginsenoside F4.

A more detailed method of preparing the ginseng flower extract is as follows.

In the present invention, the term " ginseng flower extract " means a product obtained by sequentially performing an enzyme treatment and a steaming treatment and extracting ginseng flowers.

First, the variety of ginseng used in the production of ginseng flower extract is not particularly limited, but it is preferable to use at least one selected from blue and green ginseng.

According to a preferred embodiment of the present invention, the ginseng flower may be treated with an enzyme, wherein the use of a pectinase and a cellulase improves the production of ginsenoside F4 desirable.

Preferably, the enzymatic treatment is performed by adding 0.1-1.0% by weight of pectinase and 0.5-1.0% by weight of cellulase to the ginseng flower, and then performing the reaction at 37-50 ° C for 3-7 days. If the pectinase and the cellulase are added to the ginseng flower in an amount of less than 0.1 wt% and less than 0.5 wt%, respectively, the content of the enzyme is relatively low, which is not effective enough to realize the function of the enzyme , It is uneconomical because it can exhibit sufficient efficacy without adding more than the above amount.

In addition, when the enzyme is treated, the reaction is carried out at 37 to 50 ° C for 3 to 7 days in view of maximizing the content of ginsenoside F4. If the temperature and time are out of the above range, most of the saponin is undesirably monodispersed.

As described above, the ginseng flower is enzymatically treated and then dried. The method of drying the ginseng flower is not particularly limited. However, it is preferable to carry out drying by hot air drying at 55 to 65 ° C for 48 to 72 hours to prevent corruption, Do.

Next, the dried ginseng is matured. It is preferable that the water vapor is directly performed at a pressure of 5 to 10 kgf / cm ² in terms of expanding the surface of the tissue by making the tissue porous. If the pressure is less than 5 kgf / cm ² , the surface of the tissue can not be increased due to insufficient expansion of the tissue. If the pressure exceeds 10 kgf / cm ² , And harmful components may be produced.

The ginseng flower is extracted with ethanol (EtOH) to prepare an extract, and fractionated using HP20 porous resin and ethanol (EtOH) to obtain a final ginseng flower extract.

As described above, when an extract is prepared by treating ginseng flower with enzyme and steamed treatment, ginsenoside having low bioavailability contained in ginseng is converted to ginsenoside F4 having high bioavailability, It can be applied as a cosmetic composition when the ginseng flower extract is applied to a cosmetic base as an effective ingredient.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not to be construed as limiting the scope of the present invention. It will be self-evident.

Example 1 Preparation of Ginseng Flower Extract

Example 1

After the flowers of Panax ginseng CA Meyer were dried, 0.1% by weight and 0.5% by weight of pectinase and cellulase were added, respectively, and the mixture was reacted at 50 ° C for 72 hours. Lt; / RTI > for a period of time. And steamed for 2 hours using dry steam. Extracts were prepared by extracting the cooked ginseng flower with 70% ethanol (EtOH) at 80 ℃ for 3 hours. The extracted extract was loaded on a column packed with HP20 and purified by sequentially adding ethanol (EtOH) to each concentration (40%, 50%, 65%). Then, the solution eluted at 65% The extracts were concentrated and powdered to prepare final ginseng flower extracts.

<Comparative Examples 1 to> Preparation of ginseng flower extract

Comparative Example 1

Ginseng ( Panax ginseng CA Meyer ) flowers were extracted and concentrated by ethanol (EtOH) at 80 ℃ for 3 hours to produce ginseng flower extract.

. Test Example 1 High Performance Liquid Chromatography (HPLC)

5 g of water-saturated butanol was added to 1 g of the ginseng flower extract prepared in Example 1 and Comparative Example 1, followed by mixing. The supernatant of the butanol mixture was taken and concentrated under reduced pressure. The resulting concentrate was dissolved in methanol (HPLC grade), filtered through a 0.2 μm membrane filter, and used as a sample for HPLC analysis. HPLC analysis was performed on a C ₁₈ column (3.0 x 50 mm, ID 5 μm) at UV 203 nm. The mobile phase was graded using Acetonitrile (solvent A) and water (solvent B) and separated at a flow rate of 1.6 ml / min. Table 1 shows the gradient conditions of the mobile phase.

Time (min) Moblie phase Acetonitrile (solvent A) Water (solvent B) 0.00 17 83 6.00 17 83 9.00 23 77 16.50 23.5 76.5 19.00 31 69 29.00 45 55 31.00 47 53 33.00 90 10 34.00 90 10 35.00 17 83 37.00 17 83

The results of the ginsenoside component changes of Example 1 and Comparative Example 1 are shown in Table 2 and FIG.

Example 1 (unit: mg / g) Comparative Example 1 (unit: mg / g) F4 82.5 N.D. Re N.D. 22.2

As a result, as shown in Table 2, the content of ginsenoside F4 in Example 1 and Comparative Example 1 was analyzed. As a result, it was found that the ginseng flower extract of Example 1 was superior to the ginsenoside extract of Comparative Example 1 (Ginsenoside) F4 content was remarkably high.

It can be seen that when the ginseng is enzymatically and matured, the content of ginsenoside ㄲ Re is decreased and the ginsenoside F4 is increased, which is reduced by enzymes and physical processing to give ginsenoside ) Is switched.

&Lt; Test Example 2 > Cytotoxicity test

The cytotoxicity of the ginseng flower extract obtained from Example 1 and Comparative Example 1 was measured.

First, human keratinocyte HaCat (ACTT, CLS 300493, USA) was counted in the same manner as that of a 96-well plate at a density of 2.0 × 10 ⁴ cells / well using a Hemacytometer, Of carbon dioxide for 24 hours. The cell culture obtained after 24 hours of culture was removed, and the ginseng extract obtained in Example 1 and Comparative Example 1 was prepared at 0, 10, 25, 50, 100, and 200 ug / ml. 1 μl of each concentration was mixed with the DMEM medium, and then treated for each concentration in each well. The cells were cultured at 37 ° C under 5% carbon dioxide for 24 hours. Then, a solution of MTT [3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide] per well was added at a concentration of 5 mg / ml and further cultured for 4 hours under the same conditions. After 4 hours, the MTT solution was removed and 200 μl of DMSO (dimethyl sulfoxide) solution was added to dissolve the blue formazan. Then, absorbance was measured at 570 nm using a Multi-reader (Biotec, synergy). As a control group, the absorbance was measured using distilled water instead of the sample.

The cell survival rate can be expressed by the following equation (1), and the results are shown in Table 5 and FIG.

[Equation 1]

Cell survival rate (%) = (absorbance of experimental group / absorbance of control group) X 100

Cell viability (%) Concentration (ug / ml) 0 10 25 50 100 sample Comparative Example 1 100 99.21 99.51 99.32 99.24 Example 1 100 101.52 105.43 101.78 100.20

As a result, as shown in Table 3 and FIG. 1, the concentration of ginseng flower extract was not cytotoxic even at the highest concentration of 200 ug / ml, indicating that it can be used as a safe raw material without toxicity.

<Test Example 3> Antioxidant efficacy test: DPPH radical scavenging activity

DPPH (2,2-Diphenyl-1-picrylhydrazyl) scavenging activity of the ginseng flower extract obtained from Example 1 and Comparative Example 1 was measured.

Free radicals generally act as a pathologic factor that causes the skin to dry out by destroying the skin moisturizing layer and reacting to external stimuli, which is sensitive to external stimuli. The antioxidant activity of the extract of ginseng complex according to the present invention can be confirmed by free radical scavenging ability through DPPH experiment.

In order to examine the antioxidative effect of the ginseng extract according to the present invention, specifically, the ginseng flower extracts obtained in Comparative Example 1 and Example 1 were diluted to various concentrations to prepare samples, and 100 μM DPPH (1 , 1-diphenyl-2-picrylhydrazyl) were mixed and reacted at 4 ° C for 30 minutes. Then, the amount of DPPH was measured at 520 nm in a 96-well plate. As a control, the free radical scavenging activity (%) of the sample was calculated by the following equation (2). The degree of antioxidation by DPPH free radical scavenging assay was expressed by Equation 2 and the results of antioxidative efficacy are shown in Table 4 and FIG.

&Quot; (2) &quot;

Free radical scavenging ability (%) = 100- (absorbance of the group treated with the sample / absorbance of the group not treated with the sample X 100)]

Free radical sacvening (%) Concentration (ug / ml) 0 10 25 50 100 sample Comparative Example 1 0.54 12.08 28.33 40.92 50.38 Example 1 0.48 29.30 54.44 88.71 93.67

As a result, as shown in Table 4 and FIG. 2, when the ginseng extract was treated at different concentrations, the DPPH scavenging activity was significantly higher in Example 1 than in Comparative Example 1, indicating a high antioxidative activity.

<Test Example 4> Wrinkle-improving efficacy test: MMP-1 inhibitory activity

The inhibitory activity of MMP-1 (Matrix Metallo peptidase-1) of the ginseng flower extract obtained from Example 1 and Comparative Example 1 was measured. At this time, the amount of mRNA expression was examined by RT-PCR.

Human fibroblasts were inoculated into each well of a 6-well plate to be 2 × 10 ⁵ cells and cultured in a 5% CO ² environment at 37 ° C. for 24 hours. When confluency of about 80% was reached, the cells were washed 3 times with phosphate buffered saline (PBS) and irradiated with ultraviolet light to induce skin aging. At this time, ultraviolet rays were irradiated with a total of 6.5 mJ / cm 2 using an ultraviolet A lamp (Sankyo Denki FL8BL lamp, Sankyo Denki Co., Japan). Immediately after UV irradiation, the cells were washed with phosphate buffer, and the ginseng extract obtained in Example 1 and Comparative Example 1 was added to DMEM medium containing no fetal bovine serum at concentrations of 0, 10, 25, 50, 100, and 200 ug / ml And the cells were further cultured for 24 hours. RNA was isolated using the sample-treated cells, and the RNA was isolated using an RNA isolation kit (intron). CDNA was prepared by adding MuLV reverse transcriptase and 0.5 μg of 1 mM dNTP to the obtained RNA, and the amount of expression was analyzed by real-time PCR amplification.

 The results of MMP-1 inhibitory activity are shown in Table 5 and FIG.

MMP-1 inhibition (%) Concentration (ug / ml) 0 10 25 50 100 sample Comparative Example 1 0.50 18.24 24.06 29.96 36.54 Example 1 0.50 39.6 52.84 61.54 89.82

As a result, when Example 1 and Comparative Example 1 were treated by concentration as shown in Table 5 and FIG. 3, the expression of MMP-1 increased by ultraviolet irradiation was remarkably decreased in Example 1 compared to Comparative Example 1 It was confirmed that there was an effect of improving skin wrinkles.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereto will be. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (7)

Treating the ginseng flower with 0.1 to 1.0% by weight of pectinase and 0.5 to 1.0% by weight of cellulase, reacting at 37 to 50 ° C for 3 to 7 days, and then drying;
Boiling the dried ginseng flower; And
(G) extracting ginseng flower with ethanol (EtOH) to prepare an extract, and fractionating the extract with HP20 porous resin and ethanol (EtOH) to extract ginsenoside F4 A method for producing a flower extract.
delete delete delete A ginseng flower extract prepared by the method of claim 1.
6. The ginseng flower extract according to claim 5, wherein the ginseng flower extract has an effect on skin antioxidation and wrinkle improvement.
A cosmetic composition comprising the ginseng flower extract of claim 5.

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