KR102110505B1 - Extract of ginseng for enriched chlorogenic acid and ginsenoside - Google Patents

Abstract

The present invention relates to a ginseng sprout extract having increased contents of chlorogenic acid and ginsenoside. The ginseng extract of the present invention increases contents of chlorogenic acid and ginsenosides, especially, contents of ginsenoside Re, ginsenoside Rg1, ginsenoside Rb1, or ginsenoside Rd among the ginsenosides, thereby being used as a functional material of the health functional food industry and the cosmetic industry.

Description

Sprout extract with increased chlorogenic acid and ginsenoside content {Extract of ginseng for enriched chlorogenic acid and ginsenoside}

The present invention relates to a sprout extract of chlorogenic acid (chlorogenic acid) and ginsenosides (ginsenoside) is increased.

Plants synthesize phytoalexin, a component that triggers the defense of plants by various external stimuli. Known as a plant's secondary metabolite or functional component, this substance is often promoted by temporary stress. When the plant is largely divided into the upper part and the lower part, the upper part is affected by light (light quality, light intensity, photoperiod, etc.), temperature, humidity, CO2, atmosphere (air volume, wind direction, wind speed), and other external materials. It is known that it is affected by culture, moisture (temperature, concentration), oxygen, and other foreign substances. Other biological factors include insect stress and physiological changes in the body.

Normally, plants are cultivated in the open field, but there is a problem in that functional components are enhanced by various environmental factors or it is difficult to cultivate crops of constant quality. In a closed hydroponic cultivation system such as smart farm, which is a popular cultivation technology, it is possible to cultivate crops containing a specific desired component by blocking external factors and managing desired environmental factors.

As income increases, people's well-being and demand for natural-derived materials increase, and efforts to obtain as many functional ingredients as possible from plants are required. However, it is very difficult to satisfy optimal production conditions and extraction conditions in which various specific components in a crop are high.

In Korean Patent Publication No. 10-2014-0123212 (2014.10.22), a method of increasing ginsenoside content of ginseng root using LED irradiation is described. In the Republic of Korea Patent Publication No. 10-2015-0021336 (2015.03.02), it is described with respect to the method of rapid cultivation of sprouts.

The present invention is to provide a ginseng extract having an improved content of a specific component compared to the existing no-cultivated ginseng extract, and among others, to provide optimal conditions for enhancing the content of chlorogenic acid and ginsenoside. .

The present invention is 'LED light source A combined with red light 62% or more and 87% or less, blue light 17% or more and 30% or less,' or 'LED light source B combined with red light 0% or more and 13% or less, blue light 87% or more and less than 100%' Alternatively, the ginseng cultivated is extracted by irradiating 'LED light source C combined with 100% blue light' to provide ginseng extract.

In the present invention, the ginseng extract, preferably chlorogenic acid, ginsenoside Re (Ginsenoside Re), ginsenoside Rg1 (Ginsenoside Rg1), ginsenoside Rb1 (Ginsenoside Rb1) and ginsenoside Rd (Ginsenoside Rd) It is better to improve the content than ginseng before LED irradiation.

In the present invention, the ginseng extract, preferably contains chlorogenic acid 0.24mg / g based on dry weight, ginsenoside Re (Ginsenoside Re) contains 36.98 ± 1.60mg / g based on dry weight, ginsenoside Rg1 (Ginsenoside Rg1) contains 12.92 ± 0.48mg / g on a dry weight basis, Ginsenoside Rb1 contains 11.95 ± 0.18mg / g on a dry weight basis, and Ginsenoside Rd (Ginsenoside Rd) It is good to contain 25.23 ± 0.54mg / g on a dry weight basis.

In the ginseng extract of the present invention, the extraction is preferably extracted using any one solvent selected from water, ethanol aqueous solution, methanol aqueous solution.

In the ginseng extract of the present invention, the extraction is preferably extracted at a temperature of 75 to 80 ° C., using an ethanol aqueous solution containing 50 to 80% ethanol.

In the ginseng extract of the present invention, the extraction is preferably extracted for 4-10 hours.

Ginseng extract of the present invention, chlorogenic acid and ginsenosides, especially ginsenosides, ginsenoside Re (Ginsenoside Re), ginsenoside Rg1 (Ginsenoside Rg1), ginsenoside Rb1 (Ginsenoside Rb1) or ginsenoside By increasing the content of Rd (Ginsenoside Rd), it can be used as a functional material in the health functional food industry and the cosmetics industry.

1 is a photograph showing a cultivation system for each LED wavelength.
Figure 2 is a result of comparing the chlorogenic acid content of the methanol extract of sprout ginseng according to the LED wavelength.
Figure 3 is a result of comparing the ginsenoside content of the methanol extract of sprout ginseng according to the LED wavelength.
4 is a result showing the response surface curve for temperature and ethanol.
Figure 5 shows the superimposed image (superimposed image) superimposed the reaction surface formula of the extraction yield, chlorogenic acid content and ginsenosides among the dependent variables.

The present invention is 'LED light source A combined with red light 62% or more and 87% or less, blue light 17% or more and 30% or less,' or 'LED light source B combined with red light 0% or more and 13% or less, blue light 87% or more and less than 100%' Alternatively, the ginseng cultivated is extracted by irradiating 'LED light source C combined with 100% blue light' to provide ginseng extract. At this time, the extraction, it is preferable to extract using any one solvent selected from water, ethanol aqueous solution, methanol aqueous solution.

Conventional ginseng cultivated ginseng contains only the ginsenoside content of the root portion, whereas ginseng of the present invention, by finding the optimal cultivation conditions with high specific ingredients in the smart farm, preferably chlorogenic acid, ginsenoside Re (Ginsenoside Re), Ginsenoside Rg1 (Ginsenoside Rg1), Ginsenoside Rb1 (Ginsenoside Rb1), and Ginsenoside Rd (Ginsenoside Rd) were able to improve more than the content in ginseng before LED irradiation.

In particular, the ginseng extract of the present invention only grown cultivated ginseng of 1 to 2 years for a short period of 4 weeks, but the content of chlorogenic acid of ginseng grown by the cultivation method of the present invention is 4 times that of 5-year-old ginseng. It showed a high level. In addition, ginsenoside showed higher content than 5-year-old ginseng extract in all 4 species, and the sum of ginsenoside Re, Rg1, Rb1, and Rd content contained more than 4 times that of 5-year-old ginseng. I could see through. The above results can also be interpreted as finding the optimum acceleration conditions that can enhance the chlorogenic acid and ginsenoide contents.

On the other hand, in the present invention, an optimal extraction condition was found to enhance the chlorogenic acid and ginsenoid content using alcohol, a safe solvent, even in the food processing process. An ethanol aqueous solution containing 50 to 80% ethanol was used. Using, extraction at a temperature of 75 to 80 ° C. for 4 to 10 hours was derived as an optimal condition. Under these conditions, chlorogenic acid contains 0.24mg / g on a dry weight basis, Ginsenoside Re (36) on a dry weight basis contains 36.98 ± 1.60mg / g, and Ginsenoside Rg1 (Ginsenoside Rg1) Contains 12.92 ± 0.48mg / g on a dry weight basis, contains Ginsenoside Rb1 (Ginsenoside Rb1) 11.95 ± 0.18mg / g on a dry weight basis, and Ginsenoside Rd (Ginsenoside Rd) on a dry weight basis is 25.23 ± A ginseng extract containing 0.54 mg / g could be obtained.

Hereinafter, the contents of the present invention will be described in more detail through the following examples or experimental examples. However, the scope of the present invention is not limited only to the following examples and experimental examples, and includes modifications to technical ideas equivalent thereto.

[ Manufacturing example  1: Functional ingredients according to cultivation conditions High content Sprout  culture]

1 ~ 2 years old ginseng seedlings in vertical direction under the condition of LED illumination 280 ~ 320 lux by wavelength (fluorescent lamp, R100, R87, R75, R62, R50, B62, B75, B87, B100, UVB; R: red light, B: blue light) It was investigated for 18 hours a day. In the case of nutrient solution, the mineral nutrient solution in which minerals were added to the groundwater or groundwater was irrigated and harvested after 4 weeks (FIG. 1). 1 is a photograph showing a cultivation system for each LED wavelength.

R100 means that the LED to be irradiated is combined with 100% red light. R87 means that the irradiated LED is combined with 87% red light and 13% blue light (in the present invention, R87 is also indicated as 'R87: B13'). R75 means that the irradiated LED is combined with 75% red light and 25% blue light (in the present invention, R75 is also referred to as 'R75: B25'). R62 means that the irradiated LED is combined with 62% red light and 38% blue light (in the present invention, R62 is also referred to as 'R62: B38'). R50 means that the LED to be irradiated is combined with 50% red light and 50% blue light (R50 in the present invention is also indicated as 'R50: B50').

On the other hand, B62 means that the irradiated LED is combined with 62% blue light and 38% red light (in the present invention, B62 is also referred to as 'R38: B62'). B75 means that the irradiated LED is combined with 75% blue light and 25% red light (in the present invention, B75 is also referred to as 'R25: B75'). B87 means that the irradiated LED is combined with 87% blue light and 13% red light (in the present invention, B87 is also indicated as 'R13: B87'). B100 means that the LED to be irradiated is combined with 100% blue light.

[practice Yes  1: according to the cultivation conditions Sprout  Analysis of active ingredient of methanol extract]

1. Experiment Outline

The sprouts cultivated in Preparation Example 1 were lyophilized at -50 ° C or less and vacuumed for 72 hours using a freeze dryer (Ilshin Lab, Bondiro) to prepare a sample. Extracted with 80% methanol aqueous solution for chlorogenic acid analysis, and extracted with 50% methanol aqueous solution for ginsenoside analysis 22 types of ginsenosides (Rd, Rb2, Rc, Rb1, Rb3, F2, Rg3 (S), Rg3 (R), compound K, Rh2 (S), Rh2 (R), PPD, Re, Rg1, F1, Rh1 (S), Rh1 (R), Rg2 (S), Rg2 (R), Rf, PPT ( S), PPT (R)) content was analyzed using high-speed liquid chromatography.

2. According to the LED wavelength Sprout  Of methanol extract Chlorogenic acid  Content comparison

The content of chlorogenic acid in the methanol extract of sprouts was comparatively analyzed. When cultivated with different LED wavelengths, the chlorogenic acid content increased in the group treated with the majority of LEDs compared to the control group treated with fluorescent lamps. In particular, the ratio of red light and blue light was 346% in the 13:87 treatment group. Increased (Figure 2). Figure 2 is a result of comparing the chlorogenic acid content of the methanol extract of sprout ginseng according to the LED wavelength.

3. According to the LED wavelength Sprout  Comparison of the content of 22 ginsenosides in methanol extract

The content of ginsenosides in methanol extract of sprouts was comparatively analyzed. In all groups, the content of Rd was the highest among the protopanaxadiols and the content of Re was the highest among the protopanaxadiols. When cultivated with different LED wavelengths, the ginsenoside content increased in the group treated with the majority of LEDs compared to the control group treated with fluorescent lamps. In particular, Rd, Rb2, Rb3 have a ratio of red to blue light of 75: In the 25-man treatment group, 163, 139, and 150% increase, respectively, and Rc was 117% higher in the blue light treatment group. Protopanaxadiol's final metabolite, Compound K, increased by 462% in the treatment group with a red-to-blue ratio of 13:87. Total protopanaxadiol was increased by 120% in the treatment group with a ratio of red light and blue light of 38:62 compared to a control fluorescent lamp. Total protopanic triol also increased 115% (Figure 3). Figure 3 is a result of comparing the ginsenoside content of the methanol extract of sprout ginseng according to the LED wavelength.

[ Experimental Example  One: Sprout  Comparison of active ingredients between methanol extract and 5-year-old ginseng methanol extract]

The first-year-old seedlings were cultivated by irradiating LEDs of a combination of 300 lux of LED and wavelength of 'R75: B25' in the vertical direction for 16 hours a day. The nutrient solution was irrigated with mineral nutrient solution in which minerals were added to the groundwater, and harvested after 4 weeks. After extracting the cultivated ginseng and 5-year-old ginseng with 80% methanol, chlorogenic acid, ginsenoside Re, ginsenoside Rg1, and ginsenoside Rg1, ginsenosides are the active substances of sprouts. The contents of Ginsenoside Rb1 (Ginsenoside Rb1) and Ginsenoside Rd (Ginsenoside Rd) were compared and analyzed using a high-speed liquid chromatography-mass spectrometer (HPLC-TQD-MS), and the results are shown in Table 1 below.

Comparison of chlorogenic acid and 4 ginsenosides content of methanol extract of sprout ginseng and methanol extract of 5-year-old ginseng Active ingredient 5-year-old ginseng
(mg / g dry ginseng) Sprout
(mg / g dry ginseng) Chlorogenic acid 0.03 0.13 Ginsenoside Re 1.51 6.45 Ginsenoside Rg1 1.11 3.20 Ginsenoside Rb1 0.81 1.22 Ginsenoside Rd 0.12 4.08 Ginsenoside sum 3.55 14.95

As a result of the experiment, the content of chlorogenic acid was 4 times higher than that of 5-year-old ginseng. In addition, ginsenosides showed higher content than 5-year-old ginseng extract in all 4 species, and the sum of ginsenoside Re, Rg1, Rb1, and Rd contents contained more than 4 times that of 5-year-old ginseng. .

The above results can also be interpreted as finding the optimum acceleration conditions that can enhance the chlorogenic acid and ginsenoide contents.

[ Example  2: Sprout  Preparation of ethanol extract]

Methanol extraction can be extracted more effectively, but it is not utilized in the food industry due to toxicity problems. However, alcoholic beverages are safe solvents for use in food processing processes, and research on alcoholic beverages was required for practical application.

Accordingly, in the present invention, when extracting ethanol, chlorogenic acid (Chlorogenic acid), ginsenoside Re (Ginsenoside Re), ginsenoside Rg1 (Ginsenoside Rg1), ginsenoside Rb1 (Ginsenoside Rb1), gin To investigate the optimal extraction conditions of cenoside Rd (Ginsenoside Rd), a response surface analysis was performed.

Ethanol concentration (X1, 0 ~ 70%) as shown in Table 2 below; Two independent variables were coded -1, 0, and 1 for extraction temperature (X2, 50 ~ 80 ℃). According to the surface-centered composite design method, it was repeated 3 times in random order under the conditions of Table 3 below. After analyzing the contents of chlorogenic acid, ginsenoside Re, ginsenoside Rg1, ginsenoside Rb1, and ginsenoside Rd of the extract, they were converted into contents per dry weight of sprout and placed as dependent variables.

Coded variables Natural variables Minimum Maximum Minimum Maximum

Temperature (℃),

-One One 0 70 Ethanol concentration ( % ),

-One One 50 80

Example  number

Temperature (℃),

Ethanol concentration ( % ),

2-1 50 (-1) 0 (-1) 2-2 65 (0) 0 (-1) 2-3 80 (1) 0 (-1) 2-4 50 (-1) 35 (0) 2-5 80 (1) 35 (0) 2-6 50 (-1) 70 (1) 2-7 65 (0) 70 (1) 2-8 80 (1) 70 (1) 2-9 65 (0) 35 (0) 2-10 65 (0) 35 (0) 2-11 65 (0) 35 (0) 2-12 65 (0) 35 (0) 2-13 65 (0) 35 (0)

First, after cultivating sprouted ginseng in the manner of Experimental Example 1, dried and powdered, and 40 g of ethanol solvent was added to 2 g of sprouted ginseng (sprout: ethanol solvent = 1:20), Table 3 above. It was extracted for 6 hours by the same extraction method. Then, the extract was filtered with filter paper (Advantec No. 1). Thereafter, the filtrate was concentrated under reduced pressure to completely remove ethanol, and then lyophilized to be used in Experimental Example 2 below.

[ Experimental Example  2: Using the response surface analysis method by the central synthesis planning method Sprout  Comparison of extraction yield and active ingredient of extract]

The content of the active substance was analyzed using high performance liquid chromatography (HPLC). The chromatographic analysis conditions were as shown in Tables 4 to 5 below.

Equipment UltiMate 3000 / TSQ Endura (Thermo scientific) Column C18 (Phenomenex Kinetex, 2.1㎜ × 100㎜, 2.6㎛) Column temperature 40 ℃ Injection volume 3 μl Solvent A = formic acid 0.1% in DW
B = formic acid 0.1% in Acetonitrile Flow rate 0.3ml / min MRM  condition Compound RT
(min) Polarity Precursor
(m / z) Product
(m / z) Chlorogenicacid (-) 4.01 Negative 353.0 191.0 Rg1 (-) 10.18 Negative 799.4 475.3 Rg1 (-) 10.18 Negative 799.4 637.3 Re (-) 10.24 Negative 945.5 637.3 Re (-) 10.24 Negative 945.5 783.3 Re (-) 10.24 Negative 945.5 475.3 Rb1 (-) 12.9 Negative 1107.7 220.9 Rb1 (-) 12.9 Negative 1107.7 945.3 Rb1 (-) 12.9 Negative 1107.7 783.3 Rd (-) 13.91 Negative 945.3 621.3 Rd (-) 13.91 Negative 945.3 783.3

Equipment UltiMate 3000 (Thermo scientific) Column Column Inno C18 column
(Youngjin biochrome, Korea, 5㎛, 4.6 x 250㎜) Column temperature 30 ℃ Injection volume 10 μl Solvent A = TFA 0.3% in DW
B = Acetonitrile Flow rate 1ml / min UV condition 215nm

All experiments were repeated three or more times, and dependent variables were converted based on dry weight. In response surface analysis, independent variables (ethanol concentration), dependent variables for (extraction temperature) (extraction yield), (chlorogenic acid), (ginsenoside Re), (ginsenoside Rg1), (ginsenoside Rb1) , (Ginsenoside Rd), polynomial regression equation of (Ginsenoside sum) is as shown in Equation 1. The ginsenoside sum equals the sum of ginsenosides Re, Rg1, Rb1, and Rd. The unit of (extraction yield) is%, and the units of the remaining dependent variables are the same as mg / g (dry ginseng). For the response surface analysis, the rsm package of R project was used.

As a result of the experiment, the extraction yield and the content of each active ingredient are shown in Table 6 below. The reaction surface analysis method was performed using the results of Table 6, and a quadratic polynomial equation as shown in Table 7 was obtained.

Example
number*

Extraction yield (%),

Chlorogenic acid (mg / g dry extracts),

Ginsenoside Re (mg / g dry extracts),

Ginsenoside Rg1 (mg / g dry extracts),

Ginsenoside Rb1 (mg / g dry extracts),

Ginsenoside Rd (mg / g dry extracts),

Ginsenoside sum (mg / g dry extracts),

2-1 26.77 0.01 19.65 10.52 2.48 0.00 32.65 2-2 27.07 0.20 19.56 7.63 2.88 1.87 31.94 2-3 24.83 0.18 23.29 10.36 5.31 5.57 44.53 2-4 20.90 0.18 25.09 10.60 6.88 12.35 54.91 2-5 19.50 0.27 28.92 13.56 11.51 19.58 73.57 2-6 21.27 0.40 30.74 10.42 8.03 16.35 65.54 2-7 23.77 0.43 31.85 12.97 9.92 19.10 73.84 2-8 22.83 0.41 36.59 14.92 14.11 24.95 90.58 2-9 21.13 0.35 32.16 12.04 8.85 17.58 70.63 2-10 21.41 0.40 32.01 11.56 8.83 17.19 69.58 2-11 21.60 0.40 31.98 12.19 9.06 17.48 70.71 2-12 21.44 0.40 31.47 11.49 8.73 16.77 68.46 2-13 19.98 0.37 30.49 11.29 8.33 15.95 66.06

* Examples 2-1 to 2-8 were repeated the same extraction process and analysis three times.

Extraction yield Chlorogenic acid Ginsenoside Re Ginsenoside Rg1 Ginsenoside Rb1 Ginsenoside Rd Ginsenoside sum a constant 21.29 0.076 +29.52 +11.42 +8.55 +16.26 +65.75

-0.29 +0.35 *** +2.22 *** +1.22 *** +2.26 *** +3.57 *** +9.26 ***

-1.80 *** +0.04 *** +6.11 *** +1.63 *** +3.56 *** +8.83 *** +20.14 ***

0.88 * +0.14 *** +0.55 +1.17 *** +0.81 *** + 0.76 ** + 3.29 **

-1.25 * -0.04 *** -0.76 + 0.90 * +0.82 *** +0.32 +1.27

3.96 *** -0.10 -2.06 * -0.88 * -1.98 *** -5.17 *** -10.09 ***

* Significance at p <0.05 level; ** is significant at p <0.01 level; *** is significant at p <0.001 level

_: 온도(℃)_;

_: 에탄올 농도(%) ※

_: Temperature (℃) _;

_: Ethanol concentration (%)

Table 8 below shows the suitability of the response surface model for each dependent variable.

Variables

Adjusted

F-value Extraction yield 0.8380 0.8028 23.80 *** Chlorogenic acid 0.9472 0.9357 82.55 *** Ginsenoside Re 0.8701 0.8418 30.80 *** Ginsenoside Rg1 0.7977 0.7537 18.14 *** Ginsenoside Rb1 0.9927 0.9912 628.50 *** Ginsenoside Rd 0.9907 0.9874 440.79 *** Ginsenoside sum 0.9690 0.9623 143.77 ***

* Significance at p <0.05 level; ** is significant at p <0.01 level; *** is significant at p <0.001 level

Table 9 below shows the optimal extraction conditions for each item.

Temperature (℃),

Ethanol concentration (%),

Forecast * Extraction yield 64.11 1.62 26.68 Chlorogenic acid 71.34 41.77 0.378 Ginsenoside Re 55.06 16.55 24.11 Ginsenoside Rg1 69.09 45.94 12.35 Ginsenoside Rb1 58.36 26.00 10.71 Ginsenoside Rd 52.06 0.64 0.42 Ginsenoside sum 55.98 65.80 68.81

* Unit: The extraction yield is '%', and chlorogenic acid and ginsenoside are 'mg / g dry ginseng'.

Meanwhile, in the following, the results for Tables 6 to 9 will be summarized by item and described.

1. Extraction yield

)을 제외하고, 모두 P < 0.05 수준에서 유의성을 갖는 것으로 나타났다. 또한, 상기 표 8에 의하면, 추출수율의 반응표면분석 식의 결정계수가 0.8 이상이고, P < 0.001 수준에서 유의성이 인정되었다.The extracted yield was obtained by weighing the dried sprout sprout ethanol extract powder of Example 2. The extraction yield of Sprout ginseng extract is expressed by the response surface model formula presented in Table 7 above. As a result, the first term of temperature among polynomial variables (

), But all were found to have significance at the P <0.05 level. In addition, according to Table 8, the crystal coefficient of the reaction surface analysis formula of the extraction yield was 0.8 or more, and significance was recognized at the P <0.001 level.

2. Chlorogenic acid  ( Chlorogenic  acid)

)을 제외하고, 모두 P < 0.001 수준에서 유의성을 갖는 것으로 나타났다. 또한, 상기 표 8에 의하면, 클로로제닉산의 반응표면분석 식의 결정계수가 0.9 이상이고, P < 0.001 수준에서 유의성이 인정되었다.The chlorogenic acid content of the sprout sprout extract is expressed by the reaction surface model formula presented in Table 7 above. Square term of ethanol concentration among polynomial variables (

), But all were found to have significance at the P <0.001 level. In addition, according to Table 8, the crystal coefficient of the reaction surface analysis formula of chlorogenic acid was 0.9 or more, and significance was recognized at the P <0.001 level.

On the other hand, as a result of comparing the chlorogenic acid contents of Examples 2-1 to 2-13, it was found that the chlorogenic acid content increased as the ethanol concentration increased. In particular, in Example 2-7 (65 ° C, 70%), the chlorogenic acid content was the highest (Fig. 4).

3. Ginsenoside Re ( Ginsenoside  Re)

)과 온도의 제곱항(

)을 제외하고, 모두 P < 0.05 수준에서 유의성을 갖는 것으로 나타났다. 또한, 상기 표 8에 의하면, 진세노사이드 Re의 반응표면분석 식의 결정계수가 0.8 이상이고, P < 0.001 수준에서 유의성이 인정되었다.The ginsenoside Re content of the Sprout ginseng extract is represented by the response surface model formula presented in Table 7 above. In each term of the response surface model equation, the term multiplied by the ethanol concentration and temperature (crossing term;

) And the square term of temperature (

), But all were found to have significance at the P <0.05 level. In addition, according to Table 8, the crystal coefficient of the reaction surface analysis formula of ginsenoside Re was 0.8 or more, and significance was recognized at the P <0.001 level.

On the other hand, as a result of comparing the content of ginsenoside Re from Examples 2-1 to 2-13, it was observed at the highest level in Example 2-8 (80 ° C., 70%) (FIG. 4).

4. Ginsenoside Rg1  ( Ginsenoside Rg1 )

The ginsenoside Rg1 content of Sprout ginseng extract is expressed by the response surface model formula presented in Table 7 above. Each of the terms of the Banung surface model formula was found to have significance at the P <0.05 level. In addition, according to Table 8, the expression of the reaction surface analysis of ginsenoside Rg1 was recognized as significant at P <0.001 level.

On the other hand, as a result of comparing the content of ginsenoside Rg1 from Examples 2-1 to 2-13, it was observed at the highest level in Example 2-8 (80 ° C., 70%) (FIG. 4).

5. Ginsenoside Rb1  ( Ginsenoside Rb1 )

The ginsenoside Rb1 content of the bud ginseng extract is represented by the response surface model formula presented in Table 7 above. Each term of the response surface model formula was found to have significance at the P <0.001 level. In addition, according to Table 8, the crystal coefficient of the reaction surface analysis formula of ginsenoside Rb1 was 0.99 or more, and significance was recognized at the P <0.001 level.

On the other hand, as a result of comparing the content of ginsenoside Rb1 from Examples 2-1 to 2-13, it was observed at the highest level in Example 2-8 (80 ° C., 70%) (FIG. 4).

6. Ginsenoside Rd ( Ginsenoside  Rd)

)을 제외하고, 모두 P < 0.01 수준에서 유의성을 갖는 것으로 나타났다. 이것으로 보아 에탄올 농도에 의한 영향이 진세노사이드 Rd의 추출에 더 결정적이었음을 알 수 있었다. 또한, 상기 표 8에 의하면, 진세노사이드 Rd의 반응표면분석 식의 결정계수가 0.99 이상이고, P < 0.001 수준에서 유의성이 인정되었다.The ginsenoside Rd content of the bud sprout extract is expressed by the response surface model formula presented in Table 7 above. Square term of temperature among polynomial variables (

), But all were found to have significance at the P <0.01 level. From this, it was found that the influence by ethanol concentration was more decisive for the extraction of ginsenoside Rd. In addition, according to Table 8, the crystal coefficient of the reaction surface analysis formula of ginsenoside Rd was 0.99 or more, and significance was recognized at the P <0.001 level.

On the other hand, as a result of comparing the content of ginsenoside Rd1 from Examples 2-1 to 2-13, it was observed at the highest level in Example 2-8 (80 ° C., 70%) (FIG. 4).

7. Ginsenoside Re, Rg1 , Rb1 , Rd content sum

)을 제외하고, 모두 P < 0.01 수준에서 유의성을 갖는 것으로 나타났다. 이것으로 보아 에탄올 농도에 의한 영향이 진세노사이의 추출에 더 결정적이었음을 알 수 있었다. 또한, 상기 표 8에 의하면, 진세노사이드 Rd의 반응표면분석 식의 결정계수가 0.9 이상이고, P < 0.001 수준에서 유의성이 인정되었다.The ginsenoside sum of the sprout sprout extract was obtained by adding the contents of ginsenoside Re, Rg1, Rb1, and Rd. According to the reaction surface model equation in Table 7, the square term of temperature among polynomial variables (

), But all were found to have significance at the P <0.01 level. From this, it was found that the effect of ethanol concentration was more decisive for the extraction of ginsenocytosis. In addition, according to Table 8, the crystal coefficient of the reaction surface analysis formula of ginsenoside Rd was 0.9 or more, and significance was recognized at the P <0.001 level.

On the other hand, as a result of comparing the content of ginsenoside Rd1 from Examples 2-1 to 2-13, it was observed at the highest level in Example 2-8 (80 ° C., 70%) (FIG. 4).

8. Prediction of optimal extraction conditions for alcohol extraction

FIG. 5 is a superimposed image of superimposed reaction surface formulas of extraction yield, chlorogenic acid content, and ginsenosides among the dependent variables, and each dependent variable was utilized to predict a range of optimal extraction conditions that can be maximized (Table) 10). The predicted value and the experimental value are compared at 80 ° C. and 60%, which correspond to the range of the optimal conditions of FIG. 5, and are shown in Table 11 below. Under these conditions, the extraction yield was 21.11%, chlorogenic acid 0.36mg / g, ginsenoside Re 34.68mg / g, ginsenoside Rg1 15.08mg / g, ginsenoside Rb1 13.75mg / g, ginsenoside Rd 24.36mg / g, ginsenoside sum was predicted to be 87.87mg / g.

Desirable range Optimum conditions

Temperature (℃),

75-80 80 Ethanol concentration (%),

50 ~ 70 60

Condition Forecast
(mg / g dry extracts) Experimental value
(mg / g dry extracts) Extraction yield 80 ℃, 60% 21.11% 25.67 ± 0.28 Chlorogenic acid 0.36 0.24 ± 0.00 Ginsenoside Re 34.68 36.98 ± 1.60 Ginsenoside Rg1 15.08 12.92 ± 0.48 Ginsenoside Rb1 13.75 11.95 ± 0.18 Ginsenoside Rd 24.36 25.23 ± 0.54 Ginsenoside sum 87.87 87.09 ± 2.32

[ Example  3: Sprout  extract Scale up  production]

For production on an industrial scale, a sample of sprout was extracted by increasing the production scale under optimal extraction conditions. 400 g of sprouts cultivated in the manner of Experimental Example 1 was extracted using 8 L of ethanol solvent, and in order to maximize the production yield, sprouts were extracted and the remaining foil was extracted again three times. Primary extraction was performed at 80 ° C and 60% conditions corresponding to optimal extraction conditions, and secondary extraction was performed at 70 ° C and 30% conditions, and third extraction was performed at 60 ° C and 0% conditions. The extraction times were all equal to 6 hours, and a part of the extract was sampled to track the stepwise component change. The extracts of each step were all mixed and concentrated and dried to ensure the final product. The content of chlorogenic acid and ginsenoside components in each extraction step and final product are as follows. The final product weight was 147.97 g, which showed an extraction yield of 36.99% (w / w).

 step Ingredient content (mg / g dry extract) Chlorogenic acid Ginsenoside Re Ginsenoside Rg1 Ginsenoside Rb1 Ginsenoside Rd Ginsenoside sum 1st extraction 0.377 41.5 12.4 8.5 31.8 94.2 2nd extraction 0.207 27.2 8.8 5.5 21.9 63.4 3rd extraction 0.087 16.9 5.2 1.6 3.4 27.1 final 0.330 36.7 10.4 6.6 25.0 78.6

Claims (6)

It is obtained by extracting sprouted cultivated by irradiating 'LED light source combined with red light 75% and blue light 25%' to seedling ginseng at a temperature of 75 ~ 80 ℃ using 50 ~ 80% (v / v) ethanol aqueous solution,
Chlorogenic acid, Ginsenoside Re, Ginsenoside Rg1, Ginsenoside Rb1, and Ginsenoside Rd were improved over the contents in seedlings before LED irradiation. Characterized by sprout sprout extract.
The seedlings grown by irradiating `` red LED light source composed of 75% red light and 25% blue light '' to seedling ginseng are extracted at a temperature of 75 ~ 80 ℃ using 50 ~ 80% (v / v) ethanol aqueous solution,
Chlorogenic acid, Ginsenoside Re, Ginsenoside Rg1, Ginsenoside Rb1, and Ginsenoside Rd were improved over the contents in seedlings before LED irradiation. Characterized in that the production method of the sprout.
According to claim 1,
The sprout sprout extract,
Contains chlorogenic acid 0.24mg / g on a dry weight basis,
Ginsenoside Re (Ginsenoside Re) contains 36.98 ± 1.60mg / g on a dry weight basis,
Ginsenoside Rg1 (Ginsenoside Rg1) contains 12.92 ± 0.48mg / g on a dry weight basis,
Ginsenoside Rb1 (Ginsenoside Rb1) contains 11.95 ± 0.18mg / g on a dry weight basis,
Ginsenoside Rd (Ginsenoside Rd) 25.23 ± 0.54mg / g on a dry weight basis, characterized in that it contains a sprout sprout extract.
delete delete According to claim 1,
The extraction,
Sprout ginseng extract characterized in that it is extracted for 4-10 hours.

Images