KR20200062969A - Anti-stress compositon containing Rhodiola sacra extracts and preparation method thereof - Google Patents

Abstract

The present invention relates to a method for manufacturing a Rhodiola rosea extract having increased anti-stress activity and health functional food containing the Rhodiola rosea extract manufactured by the manufacturing method as an active component, wherein the method comprises the following steps: hot water extraction of Rhodiola rosea; and supercritical extraction of the hot water extract.

Description

Anti-stress compositon containing Rhodiola sacra extracts and preparation method thereof

The present invention relates to a method for preparing an anti-stress composition comprising Hong Kyeong-cheon extract as an active ingredient and a health functional food for anti-stress containing the same as an active ingredient.

Stress refers to a state of tension to cope with the stimulus applied to the mind and body. At this time, the factor that causes stress is called a stresser. In modern society, stressors exist throughout the home and at work. Stress gives the human body the ability to respond quickly to changes that occur in the future by maintaining a tense state, and stress is usually relieved by the homeostasis principle, so there is generally no need to worry. However, if a stress factor of a size that is difficult for the mind and body to act or is exposed to constant stress or repeated stress, there is a possibility of becoming chronic, and this may cause insomnia or depression, which may cause physical and mental dysfunction.

Severe stress can be treated through medical treatment methods, but temporary stress often relies on self-treatment because it is not easily accessible to medical treatment despite frequent occurrence in the general public. Anti-stress drugs, which are currently sold as pharmaceuticals, have excellent effects, but there is a problem of side effects, so they need to be managed by a doctor. Therefore, the development of health functional foods that can be consumed without any major restrictions on a daily basis and can help relieve stress is an urgent time to improve the quality of life of modern people, and as a sole ingredient, herbal medicines that can help relieve stress Development of health functional foods derived from ingredients is urgently required.

Meanwhile, when the human body is injured or stimulated, it expresses a reaction to protect itself through a chain of hypothalamic-pituitary-adrenal axis to maintain homeostasis. That is, stress stimulation induces the production and secretion of sympathetic nerves and the production and secretion of free factors (CRF), which are the hypothalamic corticosteroids through stimulation of the cerebral cortex, and CRF of the adrenal cortex stimulating hormone (ACTH) in the anterior pituitary gland. Stimulates production and secretion, and ACTH stimulates the differentiation and proliferation of adrenal cortical cells and the production secretion of corticosterone.

Hormones such as corticosteroids and adrenaline produced during this process are released into the blood to increase pulse and blood pressure, increase respiratory rate, muscle tension, increase blood flow to the brain, heart and muscles, skin, digestive organs, kidneys, Reactions such as decreased blood flow to the liver, increased sugar and fat and cholesterol levels, and increased platelet and blood clotting factors. It is used as an indicator of the degree of stress that is changed by this series of processes.

Hongkyungcheon ( Rhodiola sacra ) is a plant widely distributed in the high mountains of Europe and Asia. It is a traditional drug effective in stimulating the nervous system, reducing depression, improving work ability, and preventing altitude sickness. It is a plant with adaptability that can survive in a high altitude environment with large daily crossings, dry, low oxygen, and strong ultraviolet rays.

The main ingredients of Hong Kyung-cheon include starch, protein, fat, sugar, flavone compounds, amino acids, neglected elements, trace essential oils, as well as Salidroside, Tyrosol and glycosides. It has been studied that ingredients derived from Hong Kyeong-cheon have cosmetic uses such as whitening and convergence, anti-cancer, antioxidant, anti-aging, and liver fibrosis suppression.

On the other hand, in Korean Patent Publication No. 2014-0136763, the extract of Hong Kyung-cheon as a polar solvent or a non-polar solvent was confirmed to have anti-stress effect by inhibiting the secretion of P substance, inhibiting CGRP secretion, inhibiting histamine free, and inhibiting interferon-gamma glass. It has been developed for use as an external agent, such as Korean Patent Application Publication No. 2016-0067137, extracts Hong Kyung-cheon by cold soaking, ultrasonic, reflux, cooling, hot water, room temperature, heating, supercritical extraction methods to reduce the skin temperature stress cosmetic composition. It is disclosed. As such, Hong Kyung-Chun has been studied to be effective in anti-stress, but there is still insufficient research.

The first problem to be solved by the present invention is to provide a method for producing a Honggyeongcheon extract with enhanced anti-stress activity by extracting the Honggyeongcheon by hot water extraction and supercritical carbon dioxide extraction.

Another problem to be solved by the present invention is to provide a health functional food for anti-stress or fatigue improvement comprising the Hong Kyeong-cheon extract prepared by the above-described manufacturing method as an active ingredient.

In order to achieve the above object, the present invention, Hongkyungcheon (Rhodiola sacra) as a supercritical carbon dioxide to extract the composition at 120 to 300 bar and 30 to 50 ℃ as an active ingredient for anti-stress or fatigue improvement composition to provide.

In addition, the present invention (1) honggyeongcheon ( Rhodiola sacra ) hot water extraction step; And (2) supercritical step of extracting the hot water extract; provides a method for producing a Hongkyungcheon extract containing anti-stress activity is increased.

According to one embodiment of the present invention, the honggyeongcheon can be further performed by further comprising the step of fermenting into a strain of Aspergillus genus before hot water extraction.

The Aspergillus genus strain may be any one selected from Aspergillus kawachii , Aspergillus oryzae and Aspergillus awamori , and the fermentation is 25 to 40 It may be carried out for 6 to 72 hours at a temperature of ℃.

According to one embodiment of the present invention, the supercritical extraction may be to extract carbon dioxide as an extraction solvent at 120 to 300 bar and 30 to 50°C.

According to the present invention, an effective indicator for anti-stress activity extracted from Honggyeongcheon may be rosavin.

In addition, the present invention provides a health functional food for anti-stress or to improve fatigue, comprising as an active ingredient an extract of honggyeongcheon with increased anti-stress activity prepared by the above-described manufacturing method.

The Honggyeongcheon extract produced by the manufacturing method according to the present invention has increased anti-stress activity, has excellent fatigue-improving effect, and has no toxicity, so it can be safely used in the human body.

Hereinafter, the present invention will be described in detail.

The present invention provides a composition for anti-stress or fatigue improvement comprising an extract extracted from Hong Kyeong- cheon ( Rhodiola sacra ) hot water extract as supercritical carbon dioxide at 120 to 300 bar and 30 to 50 ℃ as an active ingredient.

According to the present invention, the hot water extract of Honggyeongcheon may be hot water extracted using dried Honggyeongcheon, raw Honggyeongcheon or Honggyeongcheon fermentation material as a raw material.

The honggyeongcheon fermentation product may be fermented honggyeongcheon as a strain of Aspergillus for 6 to 72 hours at a temperature of 25 to 40°C, and the strain of Aspergillus aspergillus kawachii , Aspergillus It may be any one selected from Aspergillus oryzae and Aspergillus awamori , and preferably Aspergillus kawachi. The extract obtained by hot water extraction of the fermented product of Honggyeongcheon fermented with the Aspergillus kawachi showed a particularly excellent stress improvement effect in an anti-stress test on mice.

Next, the present invention (1) Hong Kyeongcheon ( Rhodiola sacra ) hot water extraction step; And (2) supercritical step of extracting the hot water extract; provides a method for producing a Hongkyungcheon extract containing anti-stress activity is increased.

First, hot water is extracted from Honggyeongcheon. The honggyeongcheon may be used as it is, but it is preferable to use dried or fermented honggyeongcheon because it has less toxicity. In particular, fermented Honggyeongcheon is more preferable because its toxicity is significantly reduced and the content of the anti-stress active substance is increased.

The fermented honggyeongcheon may be fermented honggyeongcheon as a strain of Aspergillus genus. The strain of Aspergillus genus is as previously presented. The conditions for the fermentation can be achieved by performing for 6 to 72 hours at a temperature of 25 to 40 ℃. In the fermentation process, it is preferable to sterilize Hong Kyeong-cheon before fermentation with the strain. The sterilization method may be any method commonly performed in the art, for example, there may be autoclaving, autoclaving, and autoclaving methods, but is not limited thereto.

The hot water extraction may be performed at 100° C. for 0.5 to 12 hours, to improve the extraction yield and to pulverize the honggyeongcheon or/and suspend the honggyeongcheon in water to obtain an extract with an enhanced anti-stress activity effect. After treatment, hot water can be extracted. In the present invention, through the ultrasonic treatment, a hard connective tissue (rigid structural matrix) such as fibrin in Honggyeongcheon was dissociated to easily release the physiologically active component, thereby obtaining an improved yield of the active ingredient.

According to the present invention, the ultrasonic treatment may be to irradiate ultrasonic waves for 2 to 20 minutes at 20 to 35° C. under conditions of 15-25 kHz, 500-800 watt and 20 amplitude. It is difficult to expect an improvement in extraction yield when the ultrasonic irradiation time is less than the above range, and it is difficult to obtain an anti-stress activity effect. As the ultrasonic irradiation time increased, the anti-stress activity effect also tended to increase, but when it exceeded 20 minutes, the extraction yield increased significantly, but it was confirmed that the anti-stress activity enhancing effect was lowered than when the ultrasonic irradiation was not performed.

The extract of the present invention includes not only the extract by the above-described extraction solvent, but also an extract obtained by additionally performing a conventional purification process. For example, fractions obtained through various purification methods such as separation using an ultrafiltration membrane having a constant molecular weight cut-off value and separation by chromatography (size, ion, hydrophobicity or solvent affinity) are also included in the scope of the extract of the present invention. Is included.

According to the present invention, step (2) may be performed by using supercritical carbon dioxide as an extraction solvent to extract the flow rate at a rate of 5 to 20 ml/min, and extracting at 40 to 150 minutes at 30 to 50°C. It may be to be, more preferably it may be to extract at a pressure of 120 to 300 bar.

Carbon dioxide has a property close to an organic solvent in a supercritical state. As the temperature and pressure increase, not only the general non-polar properties but also the polar properties are improved, thereby exhibiting amphoteric properties, thereby improving extraction performance. According to the present invention, under the supercritical extraction conditions, there was a problem of low extraction ability at an extraction pressure of 100 bar, and when it is 350 bar or more, the extraction ability may be improved, but components that are not involved in stress improvement are also extracted, and thus substances effective for improving stress in the extract It was confirmed that the content ratio of was lowered. Therefore, it was estimated that the extraction pressure of 120 bar or 300 bar, preferably 150 to 250 bar, is a range that can maximize the effect of improving stress. According to the present invention, a representative indicator material showing an effect on the improvement of stress may be rosavin. In addition, when the extraction time is less than the above range, the content of the component exhibiting anti-stress activity is low, which is undesirable. When the extraction time exceeds the above range, the extraction yield is improved, but the components that are not involved in stress improvement are extracted together. There is.

According to the most preferred embodiment, (A) pulverizing the honggyeongcheon, (B) suspending the pulverized honggyeongcheon in water and sonication, (C) hydrolyzing the sonicated honggyeongcheon with cell wall degrading enzyme , And (D) in preparing a supercritical extract of Honggyeongcheon through the supercritical extraction of the hydrolyzed Honggyeongcheon, ① The supercritical extraction of the step (D) is (D1) primary supercritical using only supercritical carbon dioxide Critical extraction, and (D2) is performed through a process comprising the step of supplying an organic solvent to the supercritical carbon dioxide and second supercritical extraction, wherein ② methanol: hexanol: dichloromethane is 1: 0.5 to 1 : Using an organic solvent mixed in a weight ratio of 0.5 to 1, ③ is performed at the rate of 0.2 to 0.5 mL/min based on 1 L of the supercritical carbon dioxide to perform step (D2).

In the case of the Hong Kyung-cheon extract produced through such a manufacturing method, the corticosteroid content in the blood, which is one of the stress indicators, does not increase at all. It was confirmed that the coston content increased.

According to the present invention, after the hot water extraction of Honggyeongcheon according to the above-described manufacturing method, the supercritical extraction or fermentation with a strain of Aspergillus followed by the hot water extraction and supercritical extraction of the'Hongyeongcheon extract' is a rat model that is confined in a narrow cage to add restraint stress As a result of oral administration of the Honggyeongcheon extract according to the present invention, it was confirmed that the blood concentrations of cortisol, glucose, and lactic acid, which are stress indicators, decreased, and at the same time, cytotoxicity was lowered. Therefore, the Honggyeongcheon extract according to the present invention can be applied as a functional food for health promotion for anti-stress or fatigue improvement.

Health functional foods are foods made by adding natural extracts to food materials such as beverages, teas, spices, chewing gums, confectionery, etc., or by encapsulation, powdering, suspension, etc. , Unlike regular drugs, it has the advantage of not having side effects that can occur when taking drugs for a long time using food as a raw material. The health functional food of the present invention thus obtained is very useful because it can be consumed on a daily basis. The amount of Honggyeongcheon extract added to such health functional foods cannot be uniformly defined depending on the type of health functional foods to be targeted, but may be added within a range that does not impair the original taste of foods. To 50% by weight, preferably 0.1 to 20% by weight. In addition, in the case of a health functional food in the form of pills, granules, tablets or capsules, it is usually added in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight.

When the Honggyeongcheon extract according to the present invention is used as a health functional food for anti-stress or fatigue improvement, the Honggyeongcheon extract may be formulated into a powder, granule, tablet, capsule or beverage form containing the active ingredient, but is not limited thereto. However, the mixing amount of the active ingredient can be appropriately determined according to each purpose of use. For example, for formulation in the form of tablets or capsules, the active ingredient can be combined with an oral, non-toxic acceptable inert carrier such as ethanol, glycerol, water, and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrants and colorants can also be included in the mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, trachocanth or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.

As an acceptable carrier in a composition formulated as a liquid solution, as a sterile and biocompatible material, saline, sterile water, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, and one or more of these components are mixed. Other conventional additives such as antioxidants, buffers, bacteriostatic agents can be added as needed. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate pills, capsules, granules, or tablets such as aqueous solutions, suspensions, and emulsions.

In addition, the health functional food containing the Honggyeongcheon extract of the present invention as an active ingredient may be prepared using a physiologically acceptable adjuvant in addition to the active ingredient. The supplements include various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acids and salts thereof, alginic acids and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohol, carbonated beverages The carbonation agents used may be included, and these components may be used independently or in combination. The ratio of these additives is not limited, but is generally selected from 0.01 to 0.1 parts by weight based on 100 parts by weight of the total composition of the present invention.

In addition, the Honggyeongcheon extract according to the present invention can be used as a food additive that can be added to various foods. Foods that can be added include, for example, beverages, alcoholic beverages, confectionery, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, gums, ice cream, and vitamin complexes.

The food composition of the present invention may include, as an active ingredient, Honggyeongcheon extract, as well as ingredients that are commonly added during food preparation, for example, proteins, carbohydrates, fats, nutrients, seasonings, and flavoring agents. Examples of the aforementioned carbohydrates include monosaccharides, such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides, etc.; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As a flavoring agent, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizine, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is made of a drink and a beverage, in addition to the herbal medicine extract of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, and various plant extracts may be further included.

Example

Hereinafter, the present invention will be described in more detail with reference to preferred embodiments. However, these examples are intended to illustrate the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited thereby.

Example 1-1

After pulverizing the dried honggyeongcheon, hot water was extracted at 100° C. for 6 hours, then filtered and the filtrate was concentrated. The concentrated filtrate was added at a rate of 10 ml/min at a flow rate of supercritical carbon dioxide at 250 bar and 40°C, and extracted with a supercritical extraction method for 2 hours to prepare a Honggyeongcheon extract.

Example 1-2

Honggyeongcheon extract was prepared by the method of Example 1-1 using fermented Honggyeongcheon. The fermentation of Honggyeongcheon is followed by crushing the dried Honggyeongcheon, adding it to 100 g of the crushed Honggyeongcheon so that the water content is 50%, sterilizing it in an autoclave in a flask, then cooling in a clean bench, and aspergillus kawachi ( Aspergillus kawachii ) 5 g was inoculated and cultured at 28° C. for 3 days to ferment.

Example 1-3

The ultrasonic extraction method used in the present invention was extracted by generating ultrasonic waves at 20 kHz, 750 watt and 20 amplitude conditions using an ultrasonic extractor (SEEC-SONIC II, UL-Tech.com.Uiwang, Korea), and the temperature was constant. In order to keep it clean, a Circulator Water Bath was used.

After suspending the dried honggyeongcheon in water, ultrasonic waves were irradiated for 5 minutes. Next, Honggyeongcheon was fermented and hot water extracted by the method of Example 1-2 to prepare a Honggyeongcheon extract.

Example 2-1

After washing Hong Kyeongcheon and crushing, it is suspended in water and treated with ultrasonic extractor (SEEC-SONIC II, UL-Tech.com.Uiwang, Korea) for 5 minutes at 20 kHz, 750 watt and 20 amplitude. Did. In order to keep the temperature constant, a circulator water bath was used.

After removing the suspension, citric acid was added to the sonicated Honggyeongcheon to adjust the pH to 5.8. 40 g of Viscozyme L, a complex enzyme containing cellulase, beta-glucanase, hemicellulase, arabinase, and xylanase, was added to 100 g of the pulverized Honggyeongcheon pulverized product at 40°C. Enzymatic decomposition reaction was performed while homogenizing by stirring at a temperature of 200 rpm. After completion of the reaction, the enzyme was deactivated by heating at 100° C. for 10 minutes, and then the solvent was removed to prepare an enzyme decomposition product of Honggyeongcheon.

Supercritical extraction was performed primarily for 2 hours by introducing the prepared honggyeongcheon enzyme decomposition product to supercritical carbon dioxide maintained at 250 bar and 40°C. Supply of supercritical carbon dioxide to the pressurized extractor was stopped, and then an organic solvent in which methanol:hexanol:dichloromethane was mixed at a weight ratio of 1:0.75:0.75 was supplied to the pressurized extractor, but 0.4 based on 1 L of supercritical carbon dioxide. Supercritical extraction was performed at a rate of mL/min and secondarily for 2 hours to obtain a final Hongkyeongcheon supercritical extract.

Example 2-2

A supercritical extract of Honggyeongcheon was obtained in the same manner as in Example 2-1, except that only supercritical carbon dioxide was used to perform supercritical extraction first.

Example 2-3

A supercritical extract of Honggyeongcheon was obtained in the same manner as in Example 2-1, except that an organic solvent in which methanol:hexanol:dichloromethane was mixed in a weight ratio of 1: 1.5: 1.5 was used.

Example 2-4

A supercritical extract of Honggyeongcheon was obtained in the same manner as in Example 2-1, except that the organic solvent was supplied at a rate of 1 mL/min based on 1 L of supercritical carbon dioxide.

Comparative Example 1

The dried honggyeongcheon was crushed and then hot water extracted at 100°C for 6 hours to obtain a honggyeongcheon extract.

Comparative Example 2

After pulverizing the dried honggyeongcheon, the flow rate of supercritical carbon dioxide at 250 bar and 40°C was added at a rate of 10 ml/min and extracted for 2 hours to prepare a honggyeongcheon supercritical extract.

Test Example 1. Verification of the effect of inhibiting hydrogen peroxide-induced cytotoxicity

Hydrogen peroxide generates hydroxyl radicals in the cells, causing lipid peroxidation, causing cell membrane damage and cytotoxicity. It was confirmed whether the extracts according to Examples 1-1 to 1-3 and Comparative Examples 1 and 2 can inhibit the cytotoxic action by hydrogen peroxide.

The experiment was measured by MTT assay using HaCaT cells. HaCaT cells in the logarithmic growth phase were inoculated into 96-well plates at 8000 cells per well, and cultured in 200 µl of DMEM medium for 24 hours. The extracts of Examples 1-1 to 1-3 and Comparative Examples 1 and 2 were added to the medium at a final concentration of 50 µg/ml, respectively, and then reacted for 4 hours, and the cells were washed with PBS. The washed cells were incubated for 2 hours in a medium containing hydrogen peroxide at a final concentration of 1 mM to induce oxidative stress, and the cells were washed with PBS. The washed cells were reacted by incubating for another 4 hours in a medium containing 0.5 mg/ml MTT. At this time, the yellow MTT is reduced under the action of mitochondria in living cells, and changes into a purple formazan that is not soluble in water. Since MTT is attached to the cells, the culture solution was dissolved with 200 μl of a dimethyl sulfoxide (DMSO) solution to destroy the cells and release MTT. Since the amount of formazan is directly proportional to the number of living cells, the cytotoxicity was calculated by measuring the absorbance of formazan, as measured by an ELISA reader.

In addition, HaCaT cells treated with hydrogen peroxide but not added with the extract were used as controls to perform the same method as above. After measuring the absorbance of each of the samples dissolved above at 540 nm of the ELISA reader, the cytotoxicity of each solution is expressed in Table 1 below.

[Equation 1]

division Cytotoxicity (%) Control 64.4 Comparative Example 1 37.3 Comparative Example 2 30.8 Example 1-1 26.3 Example 1-2 20.1 Example 1-3 21.6

The honggyeongcheon extracts of Examples 1-1 to 1-3 had lower cytotoxicity than the control and comparative examples 1 and 2, and in particular, the fermented honggyeongcheon extract showed low cytotoxicity. On the other hand, in the case of ultrasonic irradiation, the cytotoxicity tended to increase somewhat, but it was not significant.

Test Example 2. Anti-stress activity test

Buy 30 SPF (Specific Pathogene Free) Class 6 week old male SD (Sprague-Dawley) rats weighing 190-210g and adjust for 1 week at 22ㅁ2℃, 55ㅁ10% relative humidity and 12 hours contrast cycle Solid feed and tap water were allowed to be consumed freely. Randomly divided into 5 groups of 8 animals per group.

The control group and the drug administration group were orally administered for 4 weeks once a day for 200 mg per kg of rat body weight. After 4 weeks of drug administration, it was performed in a restraint cage (DH-SCO3, Daehan Biolink) for 5 hours. In order to evaluate the anti-stress activity, blood concentrations (µg/㎗) of cortisol, glucose and lactic acid, which are stress indicators, were measured, and are shown in Table 1 below.

Normal group: No treatment group

Control: Stress-induced group after 28 days of distilled water administration

Drug administration group: Stress-inducing group after administration of 200 mg/kg Hong Kyung-cheon extract once a day for 28 days

division Normal Control Comparative Example 1 Comparative Example 2 Example 1-1 Example 1-2 Example 1-3 Cortisol 5.42 39.56 32.61 30.59 26.15 23.34 21.81 Glucose 108.31 157.64 140.81 129.16 124.29 120.02 116.23 Lactic acid 11.26 28.12 19.15 18.56 12.34 11.83 11.76

Looking at the blood concentration (㎍ / ㎗) data of the stress indicators presented in Table 2, the concentration of stress-related indicators related to the increase in stress in the blood of the group orally administered the Hong Kyeong-cheon extract according to the present invention Was low. In addition, it was confirmed that Examples 1-2 and 1-3 using Honggyeongcheon fermentation material as a raw material exhibited more anti-stress activity.

Test Example 3: Measurement of blood corticosteroid concentration

As a drug, instead of using the honggyeongcheon extracts of Examples 1-1 to 1-3 and Comparative Examples 1 and 2, except for using the honggyeongcheon extracts of Examples 2-1 to 2-4, the above Test Example 2 The test was conducted according to the criteria.

Specifically, blood is put into a test tube for serum separation, and serum is separated by centrifugation for 10 minutes at 3,000 rpm at 4°C within 2 hours after blood collection, and then stored at -70°C, and then the concentration of corticosteroid in serum is measured. And blood biochemistry. Measurement of the concentration of corticosterone was performed using a mouse corticosteroid ELISA kit (Abcam, USA) according to the manufacturer's test method, followed by a microplate reader (BioTek-EL800, BioTek Instruments, Inc.,USA) at 450 nm. The absorbance was measured at, and the concentration of corticosteroid in serum was analyzed using a standard curve prepared as the absorbance of a standard sample.

As a result, the group administered with the extract of Honggyeongcheon in Example 2-1 showed a concentration of corticosteroid in the blood of 40.98 ㅁ 31.67 ng/ml, statistically significant difference from the normal group (41.46 ㅁ 33.22 ng/ml) On the other hand, it was confirmed that the corticosteroid concentration in the blood was statistically significantly increased by about 30% to 70% in the group administered with the Honggyeongcheon extracts of Examples 2-2 to 2-4 (p<0.01). .

Claims (4)

(1) hot water extraction of Rhodiola sacra ; And
(2) supercritical extraction of the hot water extract; as a method for producing a Honggyeongcheon extract with increased anti-stress activity, comprising:
The honggyeongcheon is fermented into a strain of Aspergillus genus before hot water extraction,
The Aspergillus genus strain is any one selected from Aspergillus kawachii , Aspergillus oryzae and Aspergillus awamori ,
The fermentation is carried out at a temperature of 25 to 40 ℃ for 6 to 72 hours,
The supercritical extraction extracts carbon dioxide as an extraction solvent at 120 to 300 bar and 30 to 50 ℃,
The anti-stress activity is a method for producing a Honggyeongcheon extract with increased anti-stress activity, characterized in that Rosabin is used as an indicator. (A) grinding the honggyeongcheon,
(B) suspending the ground honggyeongcheon in water and sonication,
(C) hydrolyzing the ultrasonically treated Hong Kyeong-cheon with cell wall degrading enzyme, and
(D) a method for producing a supercritical extract of Honggyeongcheon characterized in that it comprises the step of supercritical extraction of the hydrolyzed Honggyeongcheon,
The supercritical extraction of step (D) includes (D1) primary supercritical extraction using only supercritical carbon dioxide, and (D2) secondary supercritical extraction by supplying an organic solvent to the supercritical carbon dioxide. Is done through the process of
The organic solvent is a mixed organic solvent in which methanol:ethanol:dichloromethane is mixed in a weight ratio of 1:2 to 3:0.5 to 1,
In the (D2) step, the organic solvent is prepared at a rate of 0.2 to 0.5 mL/min based on 1 L of the supercritical carbon dioxide. Claim 1 or claim 2, wherein the anti-stress honggyeongcheon extract prepared by the method according to any one of claims. A health functional food for improving stress or improving fatigue, comprising as an active ingredient an extract of Hong Kyung-cheon for anti-stress prepared by the manufacturing method according to claim 3.