KR20160088050A - Method for cultivating Astragalus membranaceus with increased antioxidant and whitening activity using light emitting plasma irradiation - Google Patents

Abstract

The present invention relates to a method for cultivating Astragalus membranaceus characterized in that antioxidant and whitening activities of Astragalus membranaceus are increased by irradiating the Astragalus membranaceus with LEP (Light emitting plasma); to Astragalus membranaceus with increased antioxidant and whitening activities cultivated by the method; to processed food comprising Astragalus membranaceus; to a skin whitening cosmetic composition comprising Astragalus membranaceus; to a health food composition for antioxidation comprising Astragalus membranaceus; and to a pharmaceutical composition for antioxidation comprising Astragalus membranaceus. The purpose of the present invention is to establish a method for cultivating Astragalus membranaceus which has improved effects in physiological activities such as antioxidant activities, tyrosinase inhibitory activities, and others compared to Astragalus membranaceus cultivated by not irradiating Astragalus membranaceus with the LEP or irradiating the same with other light sources, by irradiating Astragalus membranaceus with the LEP in order to increase functionalities such as antioxidant and whitening activities of Astragalus membranaceus in the cultivation thereof.

Description

(Method for cultivating Astragalus membranaceus with increased antioxidant and whitening activity using light emitting plasma irradiation)

The present invention relates to a method for cultivating Hwanggi, characterized in that Astragalus membranaceus is irradiated with LEP (Light emitting plasma) to enhance antioxidative and whitening activity, a method of cultivating Hwanggi, A whitening cosmetic composition, a health food composition for antioxidation, and a pharmaceutical composition for antioxidation.

Due to recent developments in medicine, society, economy and culture, the demand for modern people who want to achieve high quality of life has increased, and interest in wellness is increasing. Wellness is a combination of well-being, happiness and fitness, which means a healthy condition and means a systematic effort to maximize the potential to maintain well-being. It is a physical, mental, emotional, It is a concept that covers the area. The wellness industry is an industry that generates added value by producing and distributing products, systems and services necessary for pursuing optimal health conditions and high quality of life through active health promotion and prevention activities. In the wellness industry, the core of the well-aged industry can be summarized as anti-aging and healthy aging. As a result of estimating the wellness industry using the subcategories of the Korean Standard Industrial Classification, the total market size is about 75,980.2 billion won, which is about 7% of GDP in 2009. Among the well-aged fields, the market size is 14,895.4 billion won , 124,516 businesses, and 226,360 employees. Especially, functional cosmetics such as wrinkles and pigmentation improvement are growing, and it is an important part in the beauty industry and the demand is gradually increasing.

Due to the wellness industry, the demand for pharmaceuticals, cosmetics and food related to health is rapidly increasing, and efforts are being made to obtain the source from natural native plant resources. The active ingredients, which are widely used in cosmetics and functional food additives, are mainly synthetic substances and they have many problems such as mutagenicity, toxicity, tolerance and induction of carcinogens. Therefore, the natural antioxidants Interest has been focused. The domestic cosmetics market is worth 4 trillion won and the vegetable raw material market is about 260 billion won, which accounts for 20% of the total 7,000 species (2013, based on consumer price). Based on these social trends, we are exploring and developing natural-resource-rich ingredients that have proven functionality through scientific research such as anti-aging and skin-lightning as a raw material for natural materials in a variety of fields such as beauty and medicine, Studies have been conducted focusing on various medicinal crops that have been used as medicines since the past.

Astragalus membranaceus B. is a medicinal crop belonging to Leguminosae. It is one of the 10 most important medicinal crops among domestic medicinal crops. It is one of the most important medicinal crops in Korea, China, Japan and some parts of Europe . In Korea, it is possible to cultivate anywhere in the country. Especially, it is possible to cultivate multi - year - The main cultivation areas in Korea are around 58 provinces in Korea, among which Gangwon-do Jung-sun, Samcheok, and Jecheon, Chungbuk are main areas. Currently, Hwanggi has grown perennial plant for the annual shipment and shipment of medicinal plants for each region, but prefer annual cultivation for economic efficiency.

Hwanggi is a dormant perennial plant, whose stem is green-purple and has a height of 70 to 120 cm and many branches. From July to August, a flower stalk is raised from the axilla of the stem at the top of the stem, and the butterfly-shaped pale yellow flower blooms together and fruit is formed in the pod. Pods are dark green, mature and purple when young, completely grayish brown when ripened, about 3-5 cm in size, and 8-10 seeds. The roots of the roots stretch straight, and the outer skin is yellowish brown. When cut off, the peripheries are milky white and yellowish white.

The general composition of Hwanggi is composed of 13 g of protein per 100 g, 1 g of lipid, 35 g of crude fiber, 360 mg of calcium, 1,000 mg of phosphorus and 2,800 mg of potassium. There is a slight difference according to the number of cultivation years. Triterpenoids, isoflavonoids and polysaccharides are also known as major components, and various other components are included. The tratterpenoid component contained in Hwanggi mainly contains glycosides of saponins such as astragaloside I-IV. Isoflavonoid component is representative of formononetin and calicosin, and two components of glycoside and (6R, 11R ) 3-hydroxy-9,10-dimethoxipterocarban-3-OD-glucoside with 7 ", 2 " -dihydroxy- 3 ", 4 " -dimethoxyisoflavan- -OD-glucoside has been reported as a major component. The pharmacological effects include antioxidants, antivirals, immunity enhancing and liver function protection, and uterine contraction.

Various environmental conditions that can increase plant quality and functional ingredient content Control of the light environment during stress is known to be the most effective method for increasing functional substance content. Currently commercially available artificial light sources include Fluorescent Lamp (FL) rather than High Pressure Sodium Lamp (HPS) or Metal Halide Lamp (MH), which generate high heat due to emission and energy loss. It is mainly used. Fluorescent lamps have the advantage of being a light source for proximity lighting that does not contain much heat in the light itself, but it is advantageous to raise the lighting efficiency to a large extent. However, since the luminous efficiency is low, the brightness of light necessary for photosynthesis Photosynthetic photon flux (PPF) is lacking.

Recently, light emitting plasma (LEP) has attracted attention as an artificial light source for new plant production which can solve various problems of artificial light sources. LEP is a solid-state, high-density light source that converts power into frequency output and vaporizes the content of the device into a plasma state to produce a strong white light. In addition, it has low heat generation compared to existing artificial light sources, has high energy efficiency and high light intensity, has a small size and long life. Especially, as an artificial light source closest to the wavelength of sunlight among the artificial light sources so far, not only UV-A and UV- Of the wavelength range. However, there is a lack of research on domestic and foreign LEP as an artificial light source for plant production.

Korean Patent Laid-Open Publication No. 2012-0138949 discloses a method of cultivating cereals or plants using plasma illumination. However, there is no technology for cultivating Hwanggi, which is enhanced by antioxidation and whitening activity using LEP irradiation as in the present invention.

SUMMARY OF THE INVENTION The present invention has been made in view of the above-mentioned needs, and it is an object of the present invention to provide a method for improving the antioxidative and whitening activity of yellow moth, The present invention has been made to establish a method for cultivating Hwanggi in which the physiological activities such as antioxidative activity and tyrosinase inhibitory activity are improved as compared with the cultivated hwanggi by irradiation with different light sources.

In order to solve the above problems, the present invention provides a method for cultivating Hwanggi, which is characterized in that Astragalus membranaceus is irradiated with light emitting plasma (LEP) to enhance antioxidative and whitening activity.

The present invention also provides an antioxidant and a whitening activity-promoted erythrocyte cultivated by the above method.

Further, the present invention provides a processed food containing the hull group.

The present invention also provides a cosmetic composition for skin whitening comprising, as an active ingredient, hwanggi, the antioxidant and whitening activity of which is enhanced.

In addition, the present invention provides an antioxidant health food composition comprising, as an active ingredient, a hwanggi, the antioxidant and whitening activity being enhanced.

In addition, the present invention provides a pharmaceutical composition for antioxidant containing an antioxidant and whitening activity-enhanced hwanggi as an active ingredient.

The hwanggi grown by irradiating LEP (Light emitting plasma) of the present invention has physiological activities such as antioxidative activity and tyrosinase inhibitory activity, and total phenol and flavonoids, which are not irradiated with LEP or irradiated with other light sources, It is effective for anti-aging and whitening due to the enhancement of the same functional material content. Therefore, there is an advantageous effect that it can be advantageously used in foods, medicines, cosmetics industry,

FIG. 1 is a graph showing a measurement of light quantity per wavelength. (A) LEP wavelength, (B) photosynthetic reaction
FIG. 2 is a graph comparing the DPPH radical scavenging ability of hwanggi grown by artificial light source.
FIG. 3 is a graph comparing the total phenol contents of yellow moth grown by artificial light sources.
FIG. 4 is a graph comparing the total flavonoid contents of cultivated hwanggi by artificial light sources.
FIG. 5 is a graph comparing tyrosinase inhibitory activities of yellow moth grown by artificial light sources.
Figure 6 compares the flavonoid biosynthetic gene expression patterns of cultivated hwanggi by artificial light sources.
Con: Control: Fluorescent lamps in FIGS. 2 to 6, Red: LED red light, Blue: LED blue light, Green: LED green light, White: LED white light, LEP:

In order to accomplish the object of the present invention, the present invention provides a cultivation method of Hwanggi, which is characterized in that Astragalus membranaceus is irradiated with light emitting plasma (LEP) to enhance antioxidative and whitening activity.

In the horseradish cultivation method of the present invention, the LEP irradiation can be preferably performed for 12 to 20 hours, more preferably for 16 hours, but is not limited thereto.

The method for cultivating hwanggi in the present invention is, more specifically,

(a) seeding the seeds of Erysipelothrix seeds in a pot containing soil in a culture room at 23 to 27 ° C; And

(b) irradiating light seeded seeds of step (a) with light emitting plasma (LEP) for 12 to 20 hours,

More specifically,

(a) planting the seeds of Erigeron seeds in a pot containing soil in a culture room at 25 占 폚; And

(b) irradiating the seedlings of step (a) with light emitting plasma (LEP) for 16 hours. The cultivation of Hwanggi in the above conditions could be promoted in the hwanggi which promoted the total phenol and flavonoid contents in the hwanggi period and the functionalities such as antioxidant and whitening activity.

The present invention also provides an antioxidant and an enhancing whitening activity cultivated by the above method.

The present invention also provides a processed food containing the hull group. There is no particular limitation on the kind of the processed food. Examples of the foods to which the hwanggi can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, rice cakes, ice cream, Drinks, alcoholic beverages, and vitamin complexes, all of which are processed foods in a conventional sense.

The present invention also provides a cosmetic composition for skin whitening comprising an antioxidant and a whitening activity-enhanced hwanggi as an active ingredient. The whitening agent, which is an effective ingredient of the skin whitening cosmetic composition of the present invention, has a tyrosinase-inhibiting activity and thus can be used for skin whitening purposes and has antioxidative activity, so that it can be used for antioxidant use. The skin whitening cosmetic composition of the present invention can be used as a skin whitening cosmetic composition for skin whitening in a wide variety of applications including softening longevity, convergent lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, powder, , Body oils, and body essences, but are not limited thereto.

The present invention also provides an antioxidant health food composition comprising an antioxidant and whitening activity-promoted hwanggi as an active ingredient.

The present invention also provides a pharmaceutical composition for antioxidant containing an antioxidant and whitening activity-enhanced hwanggi as an active ingredient.

Since the antioxidative activity of the present invention has been enhanced, the composition is useful as an antioxidant for aging and various diseases caused by oxides produced by active oxygen, preferably aging, chronic alcohol poisoning, atherosclerosis, cancer, coronary heart disease, A medicament for inhibiting or treating one or more diseases selected from diabetes mellitus, Down syndrome, hepatitis, ischemia or reperfusion injury, rheumatoid arthritis, arthritis, renal failure, inflammatory reaction, various degenerative neurological diseases, stroke stroke and myocardial infarction, Health supplements, cosmetics, foods, food additives, and the like, and are not limited thereto.

When the composition of the present invention is used as a medicine, it may further comprise suitable carriers, excipients and diluents conventionally used in the preparation of pharmaceutical compositions, and may be formulated into pharmaceutically acceptable formulations according to conventional methods .

Examples of the carrier or excipient include water, dextrin, calcium carbonate, lactose, propylene glycol, liquid, paraffin, physiological saline, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gelatin, calcium phosphate, calcium Silicates, cellulose, methylcellulose, polyvinylpyrrolidone, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and minerals. These may be used alone or in combination of two or more. Both carriers and excipients can be used.

In addition, when the antioxidant composition is weakly formulated, it may further contain conventional fillers, extenders, binders, disintegrators, surfactants, anti-coagulants, lubricants, wetting agents, perfumes, emulsifiers or preservatives.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Materials and methods

1. Plant material and kind of light source

The seeds of A. membranaceus used in the present invention were cultivated at a temperature of 25 ° C in a pot containing sterilized seeds of Erysipelasthus koreana, , Blue light, green light, white light and LEP, and fluorescent light as a control. (Lutron LX-101 Lux meter) was used to measure the amount of light. As a result, a fluorescent lamp 1700lux, LED red light (650nm) 2050lux, LED blue light (450nm) 520lux, LED green light (510nm) 1780lux, LED white light ) 3555 lux, LEP (Shinkwang ITS) 2410 lux (Fig. 1).

2. Artificial light source survey

Hwanggi seeds were selected for size and weight uniformly at 4 ℃ for 8 weeks. Two filter papers were laid on a 90 × 15 mm disposable petri dish, and the embryonic seeds were plucked into 3 replicates of 50 lips. Thereafter, the culture medium maintained at 25 ° C was irradiated with six lights (fluorescent lamp, 1700 lux), LED red light (650 nm), LED blue light (450 nm), LED green light (510 nm), LED white light The condition was maintained for 16 hours and water was supplied once a day with distilled water so that the water did not dry out during germination.

3. Physiological Activity Test of Emperor of each Emperor by Artificial Light Source

1) Extraction and concentration

The upper and lower parts of Hwanggi were harvested and dried in shade for 3 days and finely cut into 3 cm. However, the amount of Hwanggi grown for 8 weeks under the condition of artificial light source was not enough and the entire plant was used for analysis. The extracted solvent was extracted with 80% ethanol for 72 hours. Filter paper (250 mm φ , Hyundai micro NO. 20) and then concentrated under reduced pressure at 45 ° C using a rotary condenser (N-1110, EYELA, Japan).

2) DPPH activity assay

DPPH is a violet compound that exhibits specific light absorption at 517 nm and is itself a very stable free radical. These radicals are very stable in organic solvents such as ethanol, and in particular, they are discolored by proton-radical scavenging among various antioxidative mechanisms, and their antioxidative activity can be easily recognized visually.

DPPH free radical scavenging method was used to measure antioxidant activity. The concentrated samples were diluted with 80% ethanol at concentrations of 100, 500, 1,000, and 2,500 ppm, respectively. After that, 100 μl of sample compound per each ppm was added to a 96-well plate, and 100 μl of 0.15 mM DPPH solution was added, followed by reaction at room temperature for 30 minutes. After the reaction, the absorbance at 519 nm was measured using a UV spectrophotometer. RC ₅₀ (㎍ / ㎖), which is a concentration that reduces the value of the control without addition of the compound by 50% after the absorbance measurement, was compared with ascorbic acid, a water soluble vitamin used as an antioxidant.

3) Total phenol content test

Folin-ciocalteau reagent, which is a method using a coloring reagent that specifically reacts with a phenolic substance and changes its color. This is a method for measuring the total phenol content, which is not a method of measuring the phenol type. As a result, 50 μl of the folin-ciocalteau reagent was added to 100 μl of the sample and stabilized for 5 minutes. 300 μl of 20% Na ₂ CO ₃ (w / v) After stabilization for 15 minutes, 1 ml of distilled water was added to measure the absorbance at a wavelength of 725 nm. Calibration curves were prepared using gallic acid as a reference material and the results are shown.

4) Total flavonoid content assay

100 μl of 10% aluminum nitrate (w / v) and 100 μl of 1 M potassium acetate (w / v) were added to 500 μl of the diluted sample by adding 900 μl of 80% Respectively. 4.3 ml of 80% ethanol was added to the mixture, and the mixture was stabilized at room temperature for 40 minutes, and absorbance was measured at a wavelength of 415 nm. Calibration curves were prepared using quercetin as a reference material and the results are shown.

5) Whitening activity test

180 μl of 0.1 M phosphate buffer solution (pH 6.8), 80 μl of 1.5 mM tyrosine solution, 40 μl of sample per artificial light source, and 40 μl of 1,250 U / ml methylation tyrosine solution were added to the test tube, followed by reaction at 37 ° C. for 15 minutes , And the absorbance at 519 nm was measured. For comparison, kojic acid was used as a positive control for the comparison.

Tyrosinase activity inhibition rate (%) = 100- (b-b ') / (a-a') 100

a: absorbance of control group

b: absorbance of test group

a ': absorbance of blank test group without addition of extract

b ': Absorbance of the blank group containing the extract

4. Analysis of active ingredients by artificial light source

1) Analysis of flavonoids

The quantitative analysis of calycosin and formononetin, major components of the flavonoids, was carried out by high performance liquid chromatography (HPLC). Samples were diluted to 1,000 ppm and analyzed by filtration with a syringe filter (0.20 μm syringe-driven filter unit, Minisart RC 15). HPLC (Shimadzu Instrument Co. LTD, Japan) equipped with a YMC-Pack ODS-AMA-303 (5, 4.6 × 250 mm, ID) column was used. The mobile phase contained solution A (water containing 0.1% glacial acetic acid) And B (acetonitrile containing 0.1% glacial acetic acid), and solution B was analyzed by concentration gradient at 15% for 0 min, 35% for 50 min, 35% for 60 min, 15% for 80 min and 15% for 90 min Respectively. The injection volume of the sample was adjusted to 20 μl and the flow rate to 1 ml per minute. The oven temperature was 40 ° C. and the wavelength of the detector was fixed at 280 nm. The calibration curves were prepared by measuring the concentrations of 5, 10, 25, 50 and 75 ㎍ / ㎖ of formalin and 5, 1.0, 2.5, 5.0 and 10.0 ㎍ / ㎖ of calicosin, respectively. And analyzed quantitatively.

2) Analysis of polyphenolic components

UHPLC (Ultra high performance liquid chromatography) was used for quantitative analysis of phenolic compounds in Hwanggi. Samples were diluted to 10,000 ppm and analyzed by filtration through a syringe filter (0.20 μm syringe-driven filter unit, Minisart RC 15). UHPLC (Thermo Fisher Scientific Inc., USA) equipped with a Thermo Accucore C18 (2.1 x 100 mm ID, 2.6 ㎛ particle size) column was used for the analysis of the phenol components. The mobile phase consisted of Solution A (water containing 0.1% glacial acetic acid) And B (acetonitrile containing 0.1% glacial acetic acid). The solution B was stored at 0% for 2 minutes, 5% for 0.5 minutes, 10% for 2.2 minutes, 15% for 5 minutes, 15.7% for 7.5 minutes, 16.6% for 8 minutes, 17.8%, 9.5 minutes, 23.9%, 14 minutes 45%, 15 minutes 100%, 15.5 minutes 100%, 16 minutes 2% and 25 minutes 2%. The injection volume of the sample was adjusted to 4 μl, the flow rate to 0.5 ml per minute, and the wavelength of the detector was fixed at 280 nm. As the reference materials used for the analysis, 23 kinds of phenol compounds were used to prepare calibration curves, and the components of the samples were quantitatively analyzed.

5. Flavonoid Biosynthesis Gene Expression Patterns by Artificial Light Source

The plant materials used in the experiment were the ground and underground parts of Hwanggi, which were grown under the six light conditions of the control (fluorescent lamp), LED blue light, LED red light, LED green light, LED white light and LEP. The sample was pulverized using liquid nitrogen, added to 500 쨉 l of trizol, allowed to mix at room temperature for 5 minutes, centrifuged at 13,000 rpm for 5 minutes, and the supernatant was transferred to a new tube. 100 μl of chloroform was added and mixed vigorously. The mixture was incubated at room temperature for 10 minutes, centrifuged at 13,000 rpm for 15 minutes, and the supernatant was transferred. The same amount of isopropanol was added and the mixture was inverted and centrifuged at 13,000 rpm The liquid was removed. The precipitated RNA was washed with 70% ethanol, and then dissolved in sterile DIPC (diethyl pyrocarbonate) -added water and used for the expression analysis.

cDNA was synthesized using M-MLV RT (moloney murine leukemia virus reverse transcriptase). 4 μl of the RNA was mixed with 1 μl of oligo (dT) primer, 1 μl of dNTP and 6 μl of DEPC-added sterilized water, and reacted at 65 ° C for 5 minutes. Then, 4 μl of 5 × first- 2 μl of 0.1 M DTT, 1 μl of RNaseOUT ^™ recombinant ribonuclease inhibitor and 1 μl (200 units) of M-MLV RT were added and incubated at 37 ° C. for 60 minutes. A total of 20 占 퐇 of the reaction solution was diluted with 60 占 퐇 of sterilized water and 1 占 퐇 was used for PCR.

The cDNA synthesized from total RNA was quantitated using an actin primer. RT-PCR conditions were denatured at 94 ° C, followed by 30 cycles of 1 min each at 94 ° C, 55 ° C and 72 ° C. Finally, The reaction was terminated by post-elongation at 72 ° C for 10 minutes. PCR amplification was performed at 94 ° C for 5 min, denaturation at 94 ° C for 1 min, annealing at 55 ° C for 1 min and elongation at 72 ° C for 1 min. After the cycles, the cells were finally elongated at 72 ° C for 10 minutes.

Example 1: DPPH activity test of Emperor Oleum by artificial light source treatment

DPPH activity assay was performed to examine the effect of artificial light source treatment on the antioxidant activity of. As a result, the lowest activity was observed in the LED green light treatment, and the activity was slightly higher than the control in all the treatments except the LED green light treatment (FIG. 2). Based on the concentration of 1 ㎍ / ㎕ of each treatment extract, it was confirmed that LEP, LED blue light, LED red light, LED white light, control (fluorescent lamp) and LED green light were superior in antioxidant ability.

Example 2: Determination of total phenol content of hwanggi by artificial light source treatment

Polyphenols are antioxidants that turn active oxygen in our bodies into harmless substances. The most widely known antioxidants are catechins in green tea, resveratrol in wine, quercetin in onion, and many isoflavones in soybeans. The phenol hydroxyl (OH) group present in the phenolic compound has a property of binding to various compounds and is known to have a physiological activity such as antioxidant, anti-cancer and antibacterial effect.

The total phenolic content was calculated by preparing a calibration curve (R ² = 0.9991) using gallic acid as a standard material (FIG. 3). The total phenolic content of Hwanggi by artificial light source was highest at 36.05 ㎎ / ㎖ in LEP treatment and lowest at 8.76 ㎎ / ㎖ in LED white light treatment. The highest value was 12.27 ㎎ / ㎖ in blue light treatment at the LED condition, but the value was almost similar.

Example 3: Determination of total flavonoid content of hwanggi by artificial light source treatment

Flavonoids are known to have high antioxidant activity by effectively eliminating active oxygen species and are known to have antiviral, anti-inflammatory and anti-cancer effects like polyphenols.

The total flavonoid content was obtained by preparing a calibration curve (R ² = 0.9975) using quercetin as a standard substance (FIG. 4). The total flavonoid contents of artificial light source were 5.94 ㎎ / ㎖ in LEP, 2.46 ㎎ / ㎖ in LED red light, and 2.44 ㎎ / ㎖ in LED white light.

Example 4: Whitening activity test of Emperor by the treatment with artificial light source

The melanin pigment which usually causes deposition of pigment and freckles is generated by polymerization reaction with amino acids and proteins via derivatives such as dopaquinone produced by the enzymatic action of tyrosine from tyrosine, A high inhibitory effect on the activity of the enzyme indicates that the whitening effect is high.

The whitening activity test of Hwanggi by artificial light source was performed by using a spectrophotometer to measure the degree of L-dopaquinone inhibition by Mushroom tyrosinase using L-tyrosine as a substrate (FIG. 5). The positive control group, kojic acid, showed 47.44%, 94.57%, and 98.87% inhibition by 100 ㎍ / ㎖, 500 ㎍ / ㎖ and 1,000 ㎍ / ㎖, respectively. The tyrosinase inhibitory activity by the light source was 65.49% at 1,000 ㎍ / ㎖ in LEP condition, and showed about two times higher inhibitory activity than the other treatments. The inhibitory activity of tyrosinase inhibition was increased with increasing treatment concentration in the treatment of LED, but the level was insufficient and the inhibitory activity was lower than that of LEP at the lowest concentration of 100 ㎍ / ㎖.

Example 5: Analysis of active ingredients of hwanggi by artificial light source treatment

1) Analysis of major flavonoid components

Calycosin and formononetin were the main flavonoid components of Hwanggi, and calicosin was found to be contained more than formalinetin in the 1 year-old roots. In this experiment, it was confirmed that the control group treated with fluorescent lamps contained more than 10 times more formalin than that of calicosin, and this aspect was also observed in the LED treatment. On the other hand, the content of formononetin was found to be 24.55 ㎎ / g in LEP, 1.7 times higher than that of other light sources, and the concentration of calicosin was 66.62 ㎎ / g, about 27.3 times (Fig. 1). In the LEP treatment, it was found to contain more calicosin than formononetin in a similar pattern to the 1-year-old rhizome (Table 1).

Flavonoid content of Hwanggi by artificial light source Light source type Formononetin (mg / g) Calicosin (mg / g) Control (Fluorescent) 14.21 1.41 Red light 14.12 3.55 Blue light 15.11 4.10 Green light 14.66 0.74 white light 14.81 2.40 LEP 24.55 66.62

2) Analysis of polyphenolic components

23 phenolic compounds were analyzed by UHPLC for the antioxidative activities of each light source (Table 2). In protocatechuic acid, it was not detected in other treatments except for LED green light (3.73 ㎍ / g) and white light (2.69 ㎍ / g) in glacial acid at LEP condition and 19.96 ㎍ / g in protocatechuic acid. Gentisic acid was highest in LEP treatment (376.64 ㎍ / g) and blue light (20.71 ㎍ / g) and white light (20.21 ㎍ / g) were higher than other LED conditions in other LED treatment conditions . In p-hydroxybenzoic acid, LEP (43.58 ㎍ / g) showed similar contents in other light sources. Chlorogenic acid and catechin were more abundant than other LED treatments. 5 times and 30 times, respectively.

Syringic acid showed higher content of white light (23.59 ㎍ / g) and LEP treatment (35.50 ㎍ / g) than vanillin. Lip treatment (128.74 ㎍ / g) 9 times higher than that of the control. (p-coumaric acid) was the most detected in LEP treatment (1435.52 ㎍ / g). Among the LED treatments, blue light (850.56 ㎍ / g) and white light (968.57 ㎍ / g) Many were detected. Ferulic acid content was higher in LEP treatment (164.00 ㎍ / g) and blue light (63.53 ㎍ / g) was higher than other LED treatments. In m-coumaric acid, LEP treatment (328.37 ㎍ / g) was found to be about 3 times higher than other light sources. Routine treatment showed LEP treatment (13,198.35 ㎍ / g) The highest content of all components was shown by the boats. o-coumaric acid was high in LEP treatment (1,055.58 ㎍ / g) and LED red light (214.82 ㎍ / g) was higher than other LED treatments.

Naringin was the highest in LEP treatment (337.80 ㎍ / g) and myricetin was the highest in LEP treatment (9,121.18 ㎍ / g). The control (2,164.80 ㎍ / g) The content was detected to be about 10 times larger than that of the sample. Resveratrol was the highest in LEP treatment (274.76 ㎍ / g) and lowest in control (4.95 ㎍ / g), while t-cinnamic acid was the lowest in LEP treatment (2,129.28 ㎍ / g) (540.22 ㎍ / g) of LED blue light treatment was the highest among the LED light source treatments. In addition, quercetin and naringenin were detected in LEP treatment, but LED white light treatment (2,141.26 ㎍ / g) was highest in camper roll. Hess ferritin was found to be slightly higher in LEP treatment (262.46 ㎍ / g) than in other light source treatments, and formalinetin, a major flavonoid component in Hwanggi, was detected in LEP treatment (296.28 ㎍ / g) Similar contents were detected. In bioanin A, LEP treatment (245.04 ㎍ / g) was the highest.

As a result, the total phenol contents of the larvae by artificial light source were found to be high in LEP treatments. Especially, the most abundant substances were Lutein (13,198.35 ㎍ / g), Quercetin (8,984.92 ㎍ / g) (14,608.65 ㎍ / g).

Content of phenolic compounds in hwanggi by artificial light source (㎍ / g) Light source type Control Red light Blue light Green light white light LEP GA 2.88
± 0.33 5.59
± 3.24 3.20
± 1.32 2.75
± 0.65 1.30
± 0.22 19.96
± 6.12 PA - - - 3.73
± 0.28 2.69
± 0.43 - GT 6.75
± 0.49 6.30
± 5.26 20.71
± 6.51 1.10
± 15.50 20.21
± 9.13 376.64
± 17.19 pH 14.05
± 0.20 12.96
± 1.41 13.39
± 0.73 10.68
± 0.16 12.06
± 1.36 43.58
± 15.08 CA 195.13
± 4.83 197.84
± 9.69 224.34
± 0.74 165.56
+ - 8.01 267.84
± 0.57 1,104.49
± 137.55 CN 48.80
± 1.70 71.43
± 21.41 74.36
± 8.54 59.78
± 0.03 74.66
± 13.69 2527.32
± 25.01 SA 2.66
± 0.78 11.93
± 0.61 15.64
± 0.21 5.16
± 0.74 23.59
+ - 3.49 35.50
± 21.36 VN 21.50
± 1.45 17.22
± 1.72 16.30
± 0.41 17.68
± 1.18 14.92
± 0.30 128.74
± 24.76 pC 393.61
± 9.95 561.90
± 34.56 850.56
± 17.64 395.14
± 11.47 968.57
± 109.09 1,435.52
± 26.44 FA 24.66
± 0.70 30.00
± 0.62 63.53
± 7.06 18.68
± 0.78 28.35
± 0.31 164.00
± 55.48 mC 125.65
± 2.51 112.26
± 0.45 105.39
± 2.65 104.09
± 0.75 138.71
± 0.46 328.37
± 39.07 RN 59.19
± 3.99 46.16
± 2.65 53.29
± 0.19 32.53
± 3.73 64.82
± 0.13 13,198.35
± 377.05 oC 99.92
± 1.48 214.82
± 1.10 84.97
± 3.86 53.10
± 5.43 80.90
± 2.59 1,055.58
± 33.10 NA 20.41
± 4.80 32.20
± 0.07 21.51
± 1.23 28.58
± 0.32 40.49
± 1.40 337.80
± 9.94 MY 2,164.80
± 1934.80 256.74
± 205.31 300.89
± 260.54 446.91
± 78.04 254.20
± 18.01 9,121.18
± 4011.28 RE 4.95
± 0.82 9.71
± 2.18 4.31
± 1.79 11.09
± 2.19 11.75
± 2.64 274.76
± 20.04 tC 260.28
± 1.04 449.57
± 8.87 540.22
± 6.95 284.04
± 2.30 437.37
± 2.35 2,129.28
± 36.34 QN 284.41
± 30.59 408.07
± 5.67 749.43
± 14.93 247.83
± 27.99 316.94
± 25.39 8,984.92
± 244.14 NG 195.07
± 4.77 361.36
± 8.82 314.59
± 5.11 382.50
± 3.68 456.19
± 18.62 14,608.65
+ - 6.91 KA 1319.25
± 1.93 1,860.25
± 95.40 1,852.29
± 14.45 1,806.27
± 19.19 2,141.26
± 80.73 547.82
± 37.17 HN 94.52
± 1.27 144.17
± 23.49 162.67
± 16.58 104.75
± 10.91 131.71
± 19.45 262.46
± 0.60 FN 13.49
± 1.82 18.20
± 1.12 15.09
± 1.30 13.41
± 1.38 11.69
± 3.44 296.28
± 17.50 BA 41.72
± 1.51 29.54
± 7.20 23.25
± 3.24 32.14
± 2.77 20.65
± 3.85 245.04
± 5.43

GA: Gallic acid, PA: Protocatechuic acid, GT: Gentisic acid, pH: p-hydroxybenzoic acid, CA: Cholrogenic acid, CN : Catechin, SA: Syringic acid, VN: Vanillin, pC: p-coumaric acid, FA: Ferulic acid, mC: m-coumarinic acid m-coumaric acid, RN: Rutin, oC: o-coumaric acid, NA: Naringin, MY: Myricetin, RE: Resveratrol, tC : t-cinnamic acid, QN: Quercetin, NG: Naringenin, KA: Kaempferol, HN: Hesperetin, FN: Formononetin ), BA: Biochanin A (Biochanin A)

Example 6: Expression pattern of gene expression of flavonoid biosynthesis of hwanggi by artificial light source

As a result of testing the expression pattern of flavonoid biosynthesis related to artificial light source of Hwanggi, it was confirmed that I3H gene, which is involved in the final product, calicosin synthesis, is expressed in LED red light, blue light and LEP treatment. Expression of PAL gene upstream of the flavonoid biosynthetic pathway was increased in LEP treatment compared to other light sources. CHS and CHI genes were expressed in all light except for green LED light, and the expression of IFS gene was increased in LED red light treatment (Fig. 6).

Claims (9)

A method of cultivating Hwanggi, characterized in that antioxidative and whitening activity is enhanced by irradiating LEP (Light emitting plasma) to Astragalus membranaceus . The method according to claim 1, wherein the irradiation is carried out for 12 to 20 hours. The method according to claim 1, wherein the antioxidant activity enhancement is an increase in total phenolic compound content or total flavonoid compound content. The method according to claim 1,
(a) seeding the seeds of Erysipelothrix seeds in a pot containing soil in a culture room at 23 to 27 ° C; And
(b) a step of irradiating light seeded seeds of step (a) with light emitting plasma (LEP) for 12 to 20 hours and growing, thereby promoting antioxidative and whitening activity. 5. An antioxidant and whitening activity promoted by the method according to any one of claims 1 to 4. A processed food containing an erythrocyte of claim 5 enhanced with antioxidant and whitening activity. A cosmetic composition for skin whitening comprising an anti-oxidant and a whitening activity-promoting hwanggi as an effective ingredient. An antioxidant health food composition containing an antioxidant and whitening activity enhancer of claim 5 as an active ingredient. A pharmaceutical composition for antioxidant comprising an anti-oxidant and a whitening activity promoted by the anti-oxidant of claim 5 as an active ingredient.

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