KR20170057923A

KR20170057923A - Composition for antioxidatant containing extract of ginseng as an active ingredient and manufacturing method thereof

Info

Publication number: KR20170057923A
Application number: KR1020150161328A
Authority: KR
Inventors: 임병우; 서경희; 조민지; 김혜지
Original assignee: 건국대학교 산학협력단
Priority date: 2015-11-17
Filing date: 2015-11-17
Publication date: 2017-05-26
Also published as: KR101828323B1

Abstract

The present invention relates to a composition for increasing antioxidant effects by comprising hot water of ginseng or an ethanol extract as active ingredients, and also relates to a manufacturing method thereof. According to the present invention, the hot water of ginseng or the ethanol extract expresses an antioxidant effect on DNA, which caused oxidative damage in vitro, so as to inhibit the oxidative damage of the DNA, and restores damaged DNA in a concentration-dependent manner.

Description

TECHNICAL FIELD The present invention relates to an antioxidative composition containing ginseng extract as an active ingredient and a method for producing the antioxidant composition and a method for producing the same. BACKGROUND ART < RTI ID = 0.0 >

The present invention relates to a composition containing hot water or ethanol extract of ginseng as an active ingredient to increase the antioxidative effect and a method for producing the same.

Oxygen is essential for human survival, but it can also act as a toxic substance. Excessive amounts of active oxygen, which is inevitably produced in the metabolic process of oxygen, disturb proteins, fats, enzymes, DNA metabolism, destroys homeostasis, and promotes disease and aging.

Reactive oxygen species (ROS) play many important roles in normal metabolism, but excessive ROS can impair protein function, cause oxidative damage to cellular macromolecules, and cause trigger cell death. The long-term state of oxidative stress is involved in the onset of various diseases such as cardiovascular disease and cancer. Oxidative damage to DNA, proteins and other macromolecules is further assumed to constitute a major type of internal damage leading to aging. Food and pharmaceutical products that behave in response to deleterious oxidation are generally supplemented with synthetic antioxidants such as BHA, BHT, TBHQ, and PG. However, demand for natural antioxidants is increasing due to long-term stability and negative consumer perception of synthetic oxidants. We found natural antioxidants with strong antioxidant activity as an alternative to biomolecular synthetic antioxidants such as vitamin C and E, phenolic compounds, carotenoids, terpenoids and plant micronutrients. Phenolic substances are one of the widely available secondary metabolites and have various structures and molecular weights. They have antioxidative and antimicrobial properties by delaying the oxidation reaction. Therefore, studies on phenolic substances, which are most of the physiologically active substances in nature, .

Ginseng (Panax ginseng C.A. Meyer) is a perennial herbaceous plant belonging to the genus Ginseng of the Araliaceae. It is a perennial plant belonging to the genus Ginseng. It is classified into ginseng, red ginseng, white ginseng and Taegeuk ginseng according to processing methods. The components of ginseng are carbohydrates (60-70%), nitrogen (12-16%), saponins (3-6%), fat-soluble components (1-2%), ash To 6%), vitamins (0.05%) and the like.

Many researches on biochemical and pharmacological actions of various components of ginseng have been carried out at present and saponin has been attracted to many researchers as an effective ingredient. Recently, it has been reported that extracts of petroleum ether (phenol ether) and phenolic antioxidant substances known to have anticancer activity in addition to saponin have been extracted from ginseng.

Ginseng contains about 30% starch, and its starch particles are round or polygonal. Red ginseng is enriched in starch of ginseng as it undergoes heat treatment at the time of boiling, and its content is yellowish brown to reddish brown by amino-carbonyl reaction. The ginseng starch begins to swell slightly at 50 ° C, reaches a maximum at 55 to 60 ° C, breaking down the micelles structure, gradually losing the ginseng starch particles, and allowing the starch to gel at 65 to 70 ° C. The initiation temperature of ginseng starch is 61 ℃ according to the amylogram measured using amylograph, and complete digestion occurs at 88 ℃. Because starch molecules in the starch are completely dissolved, the starch is susceptible to the action of starch degrading enzymes. Therefore, it is known that the starch-rich food is better in digestibility than the raw starch food. In the case of ginseng, the starch-rich red ginseng is more effective than the white ginseng in the live starch state and the digestion is absorbed well after taking it.

Red ginseng is prepared by boiling, drying and shaping the ginseng. During the heat treatment, the pharmacological effects of ginseng and white ginseng are increased by various chemical changes such as saponin, amino acid defection, browning reaction and starch hydrolysis. Studies on the effect of puffing, fermentation, high temperature and high pressure treatment on the changes of components and antioxidant activity have been reported not only in red ginseng, but also in various ginseng components according to heat treatment.

Many antioxidants in plants exist by forming covalent bonds with insoluble polymers in cell walls. Therefore, the cause of the increase of polyphenol content by heat treatment is that the cell wall of the plant is destroyed and the polyphenol component is liberated from the insoluble component. Another reason for the increase of polyphenol is that new antioxidants are formed by the production of maillard reaction products, which have antioxidant power during heat treatment and processing, and protein hydrolysis. Another reason for the increase in antioxidants such as polyphenols is that it can prevent the degradation of antioxidants caused by enzymatic oxidation in plants by heat treatment. Enzymatic antioxidants such as superoxide dismutase, glutathione peroxidase, and glucose oxidase-catalase are inactivated during the heat treatment process of food, and natural non-enzymatic antioxidants such as carotenoids, astaxanthin in fish, tocopherol in maintenance oil, And other antioxidants are formed by food processing such as hydrolysis of protein, Mailler reaction, fermentation of lactic acid, and the like.

Korean Patent Publication No. 10-2015-0046777

An object of the present invention is to provide a composition capable of inhibiting oxidative damage of DNA by using an antioxidant activity of ginseng extract by containing hot water or ethanol extract of ginseng as an effective ingredient.

In order to accomplish the above object, the present invention provides an antioxidant composition comprising hot water or an ethanol extract of ginseng as an active ingredient.

The extract may contain Phenolic, Flavonoid, and Ascorbic Acid, and the phenol content is 44 to 165 mg GAE per 100 g of dried ginseng weight, and the flavonoid content is the dry ginseng weight 28 to 43 mg CE per 100 g, and the ascorbic acid content may be 18 to 23 mg per 100 g of dried ginseng weight.

The ginseng may be ginseng processed at 115-133 DEG C and 15 lb psi.

(1) washing the ginseng with distilled water and drying it in a dryer; (2) processing the dried ginseng; And (3) a step of extracting the ginseng extract by reflux extraction of the processed ginseng. The present invention further provides a method for producing an antioxidant composition comprising the ginseng extract as an active ingredient.

The step (2) may be a treatment for 5 to 30 minutes at 115 to 133 캜 and 15 lb psi.

The step (3) may be carried out by refluxing at 85 to 100 DEG C with tertiary distilled water having a weight of 10 to 20 times the weight of ginseng or 70% ethanol.

According to the present invention as described above, the hot water or ethanol extract of ginseng exhibits an antioxidative action against DNA causing oxidative damage in vitro, thereby suppressing the oxidative damage of DNA, and restoring the damaged DNA to a concentration-dependent recovery of the extract .

Further, according to the present invention, the antioxidative activity of ginseng is increased by a simple processing method, and the antioxidant activity as well as the active ingredient are increased according to the processing time of ginseng, so that it can be used as a natural antioxidant.

1 shows a process for producing an antioxidative composition containing an extract of ginseng as an active ingredient according to an embodiment of the present invention.
FIG. 2 is a graph showing the DPPH radical scavenging activity of ginseng extract according to an embodiment of the present invention.
FIG. 3 is a graph showing the ABTS radical scavenging activity of ginseng extract according to an embodiment of the present invention.
FIG. 4 is a graph showing reduction activity of ginseng extract according to an embodiment of the present invention.
FIG. 5 is a graph showing the DNA protecting ability of the ginseng extract according to an embodiment of the present invention in accordance with oxidative damage.

Hereinafter, the present invention will be described in detail.

The antioxidative composition having an antioxidative effect according to an embodiment of the present invention contains hot water or ethanol extract of ginseng as an active ingredient. The extract may contain Phenolic, Flavonoid, and Ascorbic Acid, and the phenol content is 44 to 165 mg GAE per 100 g of dried ginseng weight, and the flavonoid content is the dry ginseng weight 28 to 43 mg CE per 100 g, and the ascorbic acid content may be 18 to 23 mg per 100 g of dried ginseng weight. The ginseng may be ginseng that has been processed at 115-133 ° C and 15lb psi for 5-30 minutes, preferably at 121 ° C, 15lb psi, and the processing may be ginseng processed for 15-30 minutes. The hot water may be tertiary distilled water, and the ethanol may be 70% ethanol.

The antioxidant composition may be used as a synthetic preservative substitute for the production of functional foods according to the intended use. In addition, they may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized preparations according to conventional methods.

When the antioxidant composition is formulated, it is prepared using a diluent such as a filler, an extender, a binder, a wetting agent, a release agent, a surfactant, or an excipient usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, and such solid preparations are formulated containing at least one excipient and the like. Lubricants such as magnesium stearate other than the simple excipients may also be included. Examples of liquid formulations for oral use include suspensions, solutions, emulsions, and syrups, and may contain various diluents such as water and liquid paraffin, which are commonly used diluents. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of suppositories include withexol, macrogol, tween 61, cacao butter, laurin, glycerol, gelatin and the like.

In addition, the present invention provides a health functional food containing the active ingredient composed of the ginseng extract and further comprising at least one extract and a nutritional ingredient and developing a natural antioxidant including a food-acceptable food-aid additive .

A method for preparing an antioxidant composition comprising ginseng extract according to one embodiment of the present invention comprises the steps of: (1) washing ginseng with distilled water and then drying with hot air in a dryer; (2) processing the dried ginseng; And (3) extracting the ginseng extract by reflux extraction of the processed ginseng.

In the step (1), hot air drying may be performed in a dryer at 40 ° C to 50 ° C for 2 days to 3 days.

The step (2) may be performed using a pressurized steam sterilizer, and is performed in a high-temperature high-pressure steam. Further, the step (2) may be processed at a temperature of 115 to 133 캜 at 15 lb psi for 5 to 30 minutes, preferably at a temperature of 121 캜 and 15 lb psi for 15 to 30 minutes.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these examples are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.

Example 1. Preparation of ginseng hot-water extract and ethanol extract

Clean the ginseng with distilled water, remove the ginseng root and the root roots, and cut the ginseng at regular intervals (0.5 cm in thickness). Drying is carried out in a dryer at 40 ° C for 2 days with hot air to remove the water branch. The dried ginseng was processed for 15 minutes or 30 minutes at 121 ° C and 15 lb psi (1.06 kg / cm 2) on the basis of 25 g of the dried ginseng, and the mixture was treated with 10 times the weight of tertiary distilled water and 70% ethanol at 85 ° C for 2 hours 3 times with reflux to obtain hot water extract and ethanol extract. Each hot water and ethanol extract was concentrated under reduced pressure at 60 ° C or less using a rotary evaporator (Eyela, Japan), and the concentrate was lyophilized at -80 ° C. The manufacturing process is shown in Fig.

Experimental Example 1. Measurement of Phenolic, Flavonoid, and Ascorbic Acid Contents

The phenolic, flavonoid, and ascorbic acid contents of the processed ginseng hot-water extract and the ethanol extract of Example 1 were as shown in Table 1 below.

The yields of hot water extract and ethanol extract of dried ginseng, processed ginseng for 15 minutes and processed ginseng for 30 minutes were 44.54%, 32.67%, 53.56%, 41.97%, 56% and 42.55% The higher the yield, the higher the yield.

Phenol content was determined as 13.67 mg GAE and 54.62 mg GAE per 100 mg of the extracts of dried ginseng and 55.82 mg GAE and 43.74 mg GAE per 100 mg of the extracted ginseng hydrothermal extract and ethanol extract, One processed ginseng hydrothermal extract and ethanol extract were 164.87 mg GAE and 76.45 mg GAE per 100 mg of extract, respectively.

The content of flavonoid was 12.11 mg CE and 44.42 mg CE per 100 mg of extracts of dried ginseng hot water extract and ethanol extract, respectively, and the processed ginseng hydrothermal extract and ethanol extract processed for 15 minutes were 37.12 mg CE and 28.35 mg CE per 100 mg of extract, respectively, One processed ginseng hydrothermal extract and ethanol extract were 42.74 mg CE and 33.97 mg CE per 100 mg of extract, respectively.

The content of ascorbic acid was determined to be 88.14 mg and 62.36 mg per 100 mg of extracts of dried ginseng hot water extract and ethanol extract, respectively, and the processed hot water extract and ethanol extract of processed ginseng for 15 minutes were 23.30 mg and 19.00 mg per 100 mg of extract, respectively. The processed hot water extract and ethanol extract of processed ginseng for 30 minutes were 21.42 mg and 17.77 mg per 100 mg of the extract, respectively.

Experimental Example 2. DPPH radical-scavenging activity

The hot water extract and the ethanol extract of the dried ginseng extracted from Example 1, the processed ginseng processed for 15 minutes, and the processed ginseng processed for 30 minutes were adjusted to a concentration of 10 mg / ml, and then 0.25, 0.5, 1, 2 and 4 mg / ml And 1 ml of each was diluted with hot water and ethanol. 10 mg / ml of dibutylhydroxy toluene (BHT) used as a standard was diluted with 1 ml of methanol, and 1 ml Respectively. The samples prepared by concentration and BHT were dispensed in a volume of 80 μl into a 96-well plate, followed by 80 μl aliquots in the order of DW, methanol, and DPPH, wrapped in foil so as not to react with light, allowed to stand at room temperature for 30 minutes , And absorbance at 517 nm was measured using an ELISA reader.

The calculation formula of DPPH radical scavenging ability is as follows.

Scavenging activity (%) = (1 - Absorbance of sample / Absorbance of control) × 100

The results are shown in FIG. 2. Referring to FIG. 2, hot water extract and ethanol extract of dried ginseng, processed ginseng for 15 minutes and processed ginseng for 30 minutes showed DPPH radical scavenging ability in a concentration dependent manner. The DPPH radical scavenging activity of dried ginseng was 17.73% in the hot water extract and 12.64% in the ethanol extract at the concentration of 2 mg / ml. The DPPH radical scavenging activity of processed ginseng for 15 minutes was 36.8% for hot water extract and 24.16% for ethanol extract at a concentration of 2 mg / ml. The DPPH radical scavenging activity of the processed ginseng for 30 minutes was 63.27% for the hot water extract and 38.87% for the ethanol extract at the concentration of 2 mg / ml.

Experimental Example 3: ABTS radical-scavenging activity

The extracts of hot ginseng extracted from Example 1, processed ginseng processed for 15 minutes, processed ginseng processed for 30 minutes and ethanol extract were mixed at a concentration of 10 mg / ml, and then 0.25, 0.5, 1, 2 and 4 mg / ml And 1 ml each. The standard Ascorbic Acid concentration was adjusted to 7, 15, 31, 62, 125 μg / ml and 0.25, 0.5, 1, 2 and 4 mg / ml. After mixing 7 mM ABTS and 2.45 mM Potassium persulphate dissolved in distilled water, the mixture is allowed to stand in a dark room for 12 to 16 hours for stabilization. This solution can be used at room temperature for 2 days. The mixture was diluted with PBS (0.01 M, pH 7.4) to 0.70 + 0.02 at 734 nm. 0.8 ml of ABTS solution and 0.2 ml of each sample were mixed with cuvette, and reacted at room temperature for 5 minutes and then measured at 734 nm on a spectrometer. Next, 0.8 ml of potassium persulphate and 0.2 ml of the standard were mixed in a cuvette. The mixture was immediately measured at 734 nm on a spectrometer, 0.8 ml of ABTS solution and 0.2 ml of DW were mixed in a cuvette, followed by reaction for 5 minutes. 734 nm. In addition, 0.8 ml of potassium persulphate and 0.2 ml of DW were mixed in a cuvette and the absorbance was measured directly at 734 nm on a spectrometer.

The scavenging activity of the ATBS radical was calculated as follows.

The results are shown in FIG. 3. Referring to FIG. 3, the hot water extract and the ethanol extract of dried ginseng, processed ginseng for 15 minutes, and processed ginseng for 30 minutes showed ABTS radical scavenging ability in a concentration-dependent manner. At the concentration of 2 mg / ml, 70.17% of dry ginseng hot water extract and 91.45% of ethanol extract were extinguished. The treated ginseng hot water extract and ethanol extract were processed in 58.79% and 89.76% for 15 minutes respectively. The ethanol extracts of the treated ginseng treated with 30 minutes of ethanol treated with ginseng extract for 30 minutes than the treated ginseng extract treated with ethanol for 30 minutes showed 87.74% and 99.43% Respectively. Both the hot and cold extracts of dried ginseng and processed ginseng processed for 15 minutes and processed ginseng for 30 minutes contain substances that act as electron donors and change free radicals as a more stable substance and terminate the radical chain reaction Especially, the efficacy was excellent in ethanol extract of processed ginseng processed for 30 minutes.

Experimental Example 4. Reducing Power

Reduction power measurements were performed with slight modifications according to the method described by Chung et al. (Chung et al. 2005). (1 ml) and 1 ml of sodium phosphate buffer (PBS) and 2.5 ml of 1% Potassium Ferricyanide were mixed at 0.25, 0.5, 1, 2, and 4 mg / The reaction mixture was transferred to a new tube after 5 ml each, and 2.5 ml of trichloroacetic acid (TCA) was added thereto. The mixture was centrifuged at 3,000 rpm for 10 minutes in a centrifuge. Add 5 ml of DW and 1 ml of ferric chloride to 5 ml of the centrifuged supernatant, and dispense 0.15 ml into a 96-well plate. Measure the absorbance at 700 nm using an ELISA reader. Effective concentrations of each sample with an absorbance value of 0.5 (EC50) were evaluated.

The results are shown in FIG. 4. Referring to FIG. 4, the reducing power of all the extracts was not more excellent than the standard, but the concentration was dependent on the concentration. In particular, the 70% ethanol extract of the processed ginseng for 30 minutes had the best reducing power, 70% ethanol extract of processed ginseng processed for 15 minutes, hot water extract of processed ginseng processed for 15 minutes, hot ginseng hot water extract and dry ginseng 70% ethanol extract were superior in reducing power.

Experimental Example 5: Prevention of oxidation DNA damege

Prevention of oxidative damage of DNA by H ₂ O ₂ was carried out with slightly modified experimental method of Tian & Hua (2005). 1 μL of plasmid PBR 322 DNA (0.5 μg / μL) was mixed with 3 μL (0.08 mM) of FeSO ₄ , 3 μL of 4 μL (v / v) 30% H ₂ O ₂ and 2 μL of different concentrations of extract Was reacted at 37 ° C for 1 hour. Then, 2 μL of 6X DNA loading dye was added and reacted to 0.8% agarose gel electrophoresis at 70 V at room temperature. DNA bands (supercoiled, linear and open circular) were stained with ethidium bromide, gels scanned through Gel documentation system (Nextep, Korea), and band quantification was performed using NEXTEP analysis software. The effect of inhibiting DNA oxidation was evaluated by the increase and decrease of supercoiled monomer based on the control value.

The results are shown in FIG. 5. Referring to FIG. 5, as a result of the protective effect of the hot water and the ethanol extract of the dried ginseng, the treated ginseng for 15 minutes, and the treated ginseng for 30 minutes against the DNA damage, plasmid DNA control Supercoiled form DNA was completely transformed into linear form due to DNA damage compared to Lane 1 (Lane 2). DNA did not have a significant protective effect when treated with hot-water extracts, whereas the DNA of Supercoiled form was restored in a dose-dependent manner upon treatment with ethanol extracts (Lanes 3 to 5). As a result, ethanol extracts showed more protective effect against DNA damage than the hot - water extracts of dry ginseng, processed ginseng treated for 15 minutes and processed ginseng treated for 30 minutes, and processed ginseng treated with dry ginseng for 15 minutes and processed No significant difference was observed between ginseng and control at concentrations of 0.5 and 1 mg / ml.

Experimental Example 6. Statistical Processing

All data are presented as mean ± standard deviation (SDM). All experiments were repeated three times. The differences between the groups were analyzed by one-way analysis of variance (ANOVA). Graphpad Prism 5 and Microsoft Excel 2007 were used for the evaluation and statistics of the graphs.

Having described specific portions of the present invention in detail, those skilled in the art will appreciate that these specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims

An antioxidant composition comprising hot water or ethanol extract of ginseng as an active ingredient.

The method according to claim 1,
Wherein the extract comprises Phenolic, Flavonoid, and Ascorbic Acid.

3. The method of claim 2,
The phenol content is 44 to 165 mg GAE per 100 g of dried ginseng weight, the flavonoid content is 28 to 43 mg CE per 100 g of dry ginseng weight, and the ascorbic acid content is 18 to 23 mg per 100 g of dried ginseng weight &Lt; / RTI >

The method according to claim 1,
Wherein the ginseng is ginseng processed at a temperature of 115 to 133 DEG C and 15 lb psi.

(1) washing the ginseng with distilled water and drying it in a dryer;
(2) processing the dried ginseng; And
(3) A method for producing an antioxidant composition comprising ginseng extract as an active ingredient, which comprises extracting ginseng extract by reflux extraction of the processed ginseng.

6. The method of claim 5,
Wherein the step (2) is carried out at a temperature of 115 to 133 캜 for 5 to 30 minutes at a rate of 15 lb psi, and the ginseng extract is used as an active ingredient.

6. The method of claim 5,
Wherein the step (3) is carried out by reflux-extraction at 85 to 100 ° C with tertiary distilled water or 70% ethanol in an amount of 10 to 20 times by weight of ginseng weight to prepare an antioxidant composition containing the ginseng extract as an active ingredient Way.