KR101592457B1 - Method for producing ginseng extract - Google Patents

Abstract

The present invention relates to a method for producing ginseng extract, and more particularly, to a method for producing ginseng extract having high pollution-free and low-cost ginsenoside content by using high hydrostatic pressure and cell wall degrading enzyme (HHP-E) .
According to the present invention as described above, it is possible to extract a high amount of various ginsenosides by a simple method, and to reduce the occurrence of contamination and reduce the cost. The present invention relates to a pharmaceutical composition containing ginsenosides not only having a high content of major ginsenosides but also minor ginsenosides and having high content of Rg1 and Rb1 and high content of Re, Rc, Rb2, Rg2, Rf, Rd, Rh1 and compound k Extract, and a health functional food, an external preparation for skin and a pharmaceutical composition containing the ginseng extract.

Description

METHOD FOR PRODUCING GINSENG EXTRACT [0002]

The present invention relates to a method for producing ginseng extract, and more particularly, to a method for producing ginseng extract having high pollution-free and low-cost ginsenoside content by using high hydrostatic pressure and cell wall degrading enzyme (HHP-E) .

Ginseng is one of the most famous herbs consumed in the world and has been used for over a thousand years in Korea and also in China and Japan (Tyler, J. Pharm. Technol. 11, 214-220, 1995) (Tonic) or as an adaptogenic agent, which has been used to improve the function of the body against various stresses, fatigue, cancer, and diabetes.

The extracts extracted from ginseng are known to have various medical effects including immunity, anticancer, antioxidant and antidiabetic activity. This effect is mainly caused by triterpenoid glucoside, which is called ginseng saponin, do. Ginsenoside is a special kind of saponin which is found in various saponins in plants. Ginsenoside has various anti-cancer, antiallergic, anti-inflammatory, mental stability, analgesic, memory, Anti-diabetic, anti-stress, improvement of respiratory diseases, digestive system improvement, blood glucose lowering, immunity enhancement, antioxidant and anti-aging.

Ginsenosides were isolated in the early 1960s and widely used in physiological and pharmacological studies. More than 40 ginsenosides with various biological activities were isolated from ginseng roots. The ginsenosides were classified as aglycon according to the structure of the aglycon as glucosides of protopanaxadiol (PPD-type ginsenosides, Ra1, Ra2, Ra3, Rb1, Rb2, Rb3, Rc, Rd and Rg3) There are three types of glycosides: protopanaxatriol, PPT-type ginsenosides, Re, Rf, Rgl, Rg2 and oleanane-type ginsenosides.

In addition, ginsenoside is a major ginsenoside consisting of Rg1 and Re of protopanaxyl triol (PPT) and Rb1, Rc, Rb2 and Rd of protopanaxadiol (PPD) (major ginsenoside) and minor ginsenosides such as Rg3, Rk, and Rg5. Minor ginsenosides may be produced by the sugar hydrolysis process of major ginsenosides and, as with the glycoside compounds present in nature, exhibit far superior effects in terms of absorption and efficacy compared to major ginsenosides.

20-O-β-D-glucopyranosyl-20 (S) -prototopanaxadiol (compound k, compound k), known as the major metabolite in the body such as the ginsenosides Rb1, Rb2, Rc, Senoside is a substance produced by elimination of the sugar moiety of saponin with the aid of digestive juices or intestinal bacteria and is not related to the decomposition of components such as Rg1 and Rb1 and is finally decomposed Respectively. Compound K is known to inhibit the proliferation of cancer cells, increase anticancer activity of anti-cancer drugs, inhibit allergies, and protect skin, and many studies have been conducted to efficiently produce them.

A known method for producing the compound K is a method in which a diol-based saponin is prepared by enzymatic digestion with NARIN or by hydrolysis, or a compound K produced as a degradation product in the large intestine after oral administration to a mouse. However, these methods have a problem that not only the yield is extremely low but also various secondary metabolites are generated, making it difficult to purify the compound K with high purity (Kohda Hiroshi and Tanaka Osamu, The Pharm. Society of Japan 95: 246 ), Karimura et al., Chem. Pharm. Bull. 38: 2859 (1990)), the method of converting ginseng saponin into compound k is mainly performed by microorganisms.

Ginsenosides are glycosides composed of a glycone and an aglycone. When ginsenosides are absorbed in the body, they are degraded and absorbed by specific microorganisms present in the intestines of the individual. In other words, ginsenoside is metabolized by intestinal microorganisms after ingestion, and its metabolites exhibit various physiological activities. Since the specific microorganisms present in the intestines are different depending on the constitution, dietary habit, etc., the conversion rate and bioavailability of ginsenoside are different, and the effect is different for each individual after taking ginseng. For example, some people absorb only the diol form of ginsenosides, some absorb the triol form, and some people do not absorb ginsenosides at all.

More specifically, the protopanaxadiol-based ginsenosides Rb1, Rb2, and Rc were produced by the intestinal bacteria of humans using 20-O- beta -D-glucopyranosyl-20 (S) -protopanaxadiol protopanaxadiol) (Hasegawa H, et al., Planta Medica 63: 463-40, 1997; Tawab MA et al., Drug Metab. Dispos. 31: 1065-71, 2003) The senosides Re and Rg1 are metabolized by intestinal bacteria to ginsenoside Rh1 or ginsenoside F1 (Hasegawa H, et al., Planta Medica 62: 453-7, 1996; Tawab MA, et al., Drug Metab Dispos. 31: 1065-71, 2003). The compounds K, Rh1 and F1 thus converted exhibit various physiological activities.

Also, the type and amount of ginsenosides contained will vary depending on the type of ginseng, the processing method, or the method of growth. Panax ginseng contains 22 PPD type ginsenosides and 14 PPT type ginsenosides whereas American ginseng (Panax quinquefolium) contains 13 PPD type ginsenosides and 5 PPT type ginsenosides . In certain species of ginseng, the composition of ginsenosides varies depending on the area such as root, stem, leaf and fruit.

Various methods have been developed for producing ginsenosides containing the physiological and pharmacological active ingredient ginsenosides as described above. These methods maximize the extraction efficiency of ginsenosides, minimize the decomposition and minimize the environmental pollution And to reduce production costs.

The interest and needs of ginseng extracts are high, and in order to enhance the efficacy of ginseng extract, ginsenosides which are easy to absorb should be extracted in a high content. Therefore, in order to produce excellent ginseng products, minor ginsenoside, which is known to have various pharmacological actions not contained in ginseng such as ginseng, as well as major ginsenosides, It is necessary to develop a low-cost, high-efficiency method that can be used.

Korean Patent No. 10-0228510 (method for producing ginsenosides Rg3 and / or Rg5) and Korean Patent Laid-Open No. 10-2013-0059773 (a health functional food containing the processed ginseng extract having the effect of improving skin wrinkles as an active ingredient) ), Korean Patent Publication No. 10-2011-0108820 (a composition for suppressing obesity containing supercritical carbon dioxide supercritical extract), and the like. However, the conventional heating and extraction method using a solvent takes a long time and low efficiency, There is a problem that it is destroyed by heat. In addition, the extraction method using ultrasonic waves has a problem that the cell membrane is broken due to the mechanical impact of the sound wave vibration and the dissolution of the contents is easy, but it is difficult to apply industrially. Supercritical extraction method using carbon dioxide is suitable for extracting lipophilic substances Extraction of water-soluble materials is limited.

As described above, the conventional ginseng extract manufacturing method suffers from various problems in industrial application due to low extraction efficiency, long time, high cost, and high energy consumption.

It is an object of the present invention to provide a method for producing a ginseng extract which is simple and low in cost compared to the conventional extraction method through application of kinds and addition ratios of glucosease having a high ginsenoside yield and ultrahigh pressure.

It is also an object of the present invention to provide a pharmaceutical composition which contains not only a high amount of major ginsenoside but also minor ginsenoside, and the ratio of Rg1 and Rb1 and the ratio of Re, Rc, Rb2, Rg2, Rf, Rd, And a health functional food, an external composition for skin, and a pharmaceutical composition containing the ginseng extract.

In order to achieve the above object, the present invention provides a method for producing a plant, comprising (1) at least one raw material selected from the group consisting of roots, stems, leaves and fruits of Panax species plants, water and a lower alcohol having 1 to 4 carbon atoms At least one solvent selected from the group consisting of: (2) The suspension prepared in the above step (1) is dissolved in a solvent selected from the group consisting of cellulase, amylase, pectinase, cellobiase and viscozyme. Preparing a mixed solution by mixing with the above-described saccharide-degrading enzyme; And (3) treating the mixed solution prepared in the step (2) at a temperature of 32 to 60 DEG C in an amount of 0.1 to 200 MPa to prepare an extract containing ginsenoside.

In step (1), the ginseng plant is selected from the group consisting of Panax ginseng, Panax quinquefolium, Panax notoginseng, Panax japonicum, Panax trifolium, Panax pseudoginseng, It is characterized by being Panax vietnamensis.

In the step (1), the suspension is mixed with 10 to 25 parts by weight of at least one raw material selected from the group consisting of root, stem, leaf and fruit of a plant of the genus Ginseng, relative to 100 parts by weight of the solvent.

In the step (2), the pH of the suspension is adjusted to 4.5 to 5 by adding a physiologically acceptable acid.

Wherein the physiologically acceptable acid is selected from the group consisting of inorganic acids including hydrochloric acid, sulfuric acid and phosphoric acid and organic acids including oxalic acid, acetic acid, succinic acid, tartaric acid, ascorbic acid, citric acid, glutamic acid, methanesulfonic acid and ethanesulfonic acid 1 or more.

In the step (2), the saccharide-degrading enzyme is mixed with 1 to 5 parts by volume of the skin of 100 parts of the suspension.

The ginseng extract is prepared by adding a mixed enzyme of pectinase and cellulase or a mixed enzyme of pectinase, cellulase, and cellulase to a glucosease digesting enzyme in step (2), and adding 95 to 105 MPa And then treating it with ultra high pressure to increase the content of total ginsenosides.

The ginseng extract is characterized in that the content of ginsenosides Rg1 and Rb1 is increased by adding amylase, cellulase, and pectinase as a saccharolytic enzyme in step (2).

In this case, the amylase, cellulase, and pectinase are added in a mixing ratio (unit / ml) of 5 to 20: 0.5 to 2: 2 to 8, and the ginseng extract is used as a functional beverage composition .

The ginseng extract is characterized in that the content of ginsenoside Rd is increased by adding a cellulase, a pectinase and a cellulase as a saccharide-degrading enzyme in the step (2).

In this case, the cellulase, pectinase and cellobiase are added in a mixing ratio (unit / ml) of 0.5: 3: 0.5 to 3: 0.5 to 3, and the ginseng extract is used as a composition for external application for skin .

The ginseng extract is characterized in that in step (2), ginsenoside Rh1 and compound k are added by adding a cellulase, an amylase and a cellulase as a saccharide-degrading enzyme.

In this case, the cellulase, amylase and cellobiase are added in a mixing ratio (unit / ml) of 0.5: 3: 0.5 to 3: 0.5 to 3, and the ginseng extract is used as a pharmaceutical composition for anti-cancer do.

The present invention provides ginseng extract extracted by the above-described method.

The present invention also provides a health functional food containing the ginseng extract as an active ingredient.

The present invention also provides a composition for external application for skin containing the ginseng extract as an active ingredient.

The present invention also provides a pharmaceutical composition for antioxidation comprising a pharmaceutically effective amount of ginsenoside contained in the ginseng extract and a pharmaceutically acceptable carrier.

The present invention also provides a pharmaceutical composition for enhancing immunity comprising a pharmaceutically effective amount of ginsenoside contained in the ginseng extract and a pharmaceutically acceptable carrier.

According to the present invention as described above, a variety of ginsenosides having a high content can be extracted by a simple method by applying the optimal type and addition rate of the saccharide enzyme and the ultrahigh pressure, thereby reducing the occurrence of contamination and reducing the cost have.

The present invention also relates to a pharmaceutical composition containing not only a high amount of major ginsenoside but also minor ginsenosides, wherein the ratio of Rg1 and Rb1 and the content of Re, Rc, Rb2, Rg2, Rf, Rd, Rh1 and compound K A ginseng extract excellent in high commerciality and a health functional food, an external preparation for skin and a pharmaceutical composition containing the ginseng extract are provided.

1 is a process for producing ginseng extract.
Figure 2 shows the content of ginsenosides extracted at various pressures.
FIG. 3 compares the total ginsenosides content of 4-year old Geumsan ginseng, 6-year old Geumsan ginseng and 4-year old ginseng ginseng treated at normal pressure (0.1 MPa) and ultra high pressure (100 MPa).
Figure 4 shows the total ginsenoside content according to the ginseng content (ginseng concentration) mixed in the solvent.
Figure 5 shows the total ginsenoside content extracted at atmospheric pressure (0.1 MPa) and ultra high pressure (100 MPa) according to various enzyme combinations. A is amylase, P is pectinase, C is cellulase, Cb is cellobiase, and V is viscose.
6 compares the yield of major ginsenosides Re + Rg1 + Rb1 + Rc + Rb2 (占 퐂 / g) according to the pH of the suspension mixed with ginseng and solvent extracted at ultra high pressure (100 MPa) and the type of cell wall degrading enzyme.
7 compares yields of major ginsenosides Re + Rg1 + Rb1 + Rc + Rb2 (占 퐂 / g) according to the ultrahigh pressure treatment temperature and cell wall degrading enzyme species extracted at ultra high pressure (100 MPa).
(D) treatment at normal pressure for 12 hours, (e) treatment at a high pressure for 12 hours at an ultrahigh pressure, (b) HPLC graph of Jinsei Nosai contained in the time-treated extract.
9 is a graph showing that the content of Rg1 + Rb1 is greatly increased when the enzyme amylase, cellulase, and pectinase are added and extracted at atmospheric pressure (0.1 MPa) and ultrahigh pressure (100 MPa).
10 compares the yields of Rd when cellulase, pectinase and cellobiase were added as enzymes and extracted at atmospheric pressure (0.1 MPa) and ultrahigh pressure (100 MPa) for 12, 18 and 24 hours, respectively.

Hereinafter, the present invention will be described in detail.

Ginseng is a perennial herb, about 60 cm in length, usually harvested 4 to 6 years later in autumn, and raw ginseng is called raw ginseng. Ginseng is classified as white ginseng and red ginseng according to the processing method and it is classified into cultivation ginseng, ginseng ginseng, and wild ginseng depending on the growing environment.

Ginseng has long been known as a natural ergogenic aid and has been shown to have various effects ranging from immunity enhancement, anti-stress, anti-cancer, anti-aging, antioxidant, hangover and tonic effects (Kim SH, et al. Biochem Pharmacol., 54 (1): 1-8, 1997; Attele AS, et al., Biochem Pharmacol., 45 (2): 178-82, 2005). , Shin et al., Cancer Causes Control, 11 (6): 565-76, 2000). It has been reported that ginsenosides Rg1 and Rb1 work in improving mitochondrial energy metabolism and promoting aereobic exercise performance (Wang LC, et al., Planta Med., 64 (2): 130- 133, 1998), it has been reported that oxidative stress is reduced by antioxidative effects of ginsenoside Rg3 and Re (Tian J, et al., Neurosci. Lett., 374 (2): 92-97 , 2005, Cho WC, et al., 2006, Eur. J. Pharmacol., 550 (1-3): 173-179, 2006).

Ginsenoside Rg1 has been shown to be effective in the enhancement of immune function (Okamura N et al. Biol. Pharm. Bull. 17 (2), 270 (1994)), antipyrotic action (Saito H et al. Japan J. Pharmacology, ), And a noxious stimulant defense action (Yoshimatsu H et al. Physiology and Behavior, 53 (1), 1 (1993)) and ginsenoside Rd has a stimulating action on adrenocortical hormone secretion (Hiai S et al. Bax. 31, 168 (1983)), ginsenoside Rb1 is a protein synthesis promoting action (Zhang JT et al. Acta Pharmaceutica Sinica 23 (1), 12 Lipid synthesis promoting action, and ginsenoside Rb2 have various pharmacological effects such as antidiabetic action, immunoregulatory action, and cholesterol metabolism.

The effects of ginseng extracts on the skin include anti-inflammatory effects (Korean J. Dermatol. 18 (1): 39-42 (1980), Korean J. Dermatol.14 (4): 335-339 (1976)), Korean J. Dermatol. 28 (4): 434-440 (1990)), acne prevention and treatment (Korean J. Dermatol.30 (1): 27-33 (1992) (Fitoterapia 57 (1): 15-28 (1986), Fitoterapia 57 (4)), antioxidant activity (Proceedings of the 2nd International Ginseng Symposium 13-17 217-222 (1986)), wound healing (Brit. J. Pharmacol. 125: 255-262 (1998), Arch Pharm Res. 25 (1): 71-76 Cells 9 (5): 476-483 (1999)), whitening (Journal of the Society of Cosmetic Scientists of Korea 27 (2): 45-56 (2001)).

PPD ginsenoside and PPT ginsenoside have different functions in the body. PPD saponin, represented by ginsenoside Rb1, has an inhibitory action on the central nervous system and is known to have mental stability, nervous relaxation, analgesia, Blood pressure lowering, and papaverine positive action. PPT saponin represented by ginsenoside Rg1 is known to have an antipyrotic action by acting excitably on the central nervous system.

Res. 9: 411-7, 1998; Lee SJ, et al., ≪ RTI ID = 0.0 > 1998). ≪ / RTI > , Cancer Lett. 144: 39-43, 1999), Rh1 is a cytotoxic effect for various cancer cell growth (Odashima S, et al., Cancer Res. 45: 2781-4, 1985; Ota T, et al., Cancer Ginseng symp. Seoul 127-31, 1993) and anti-allergic, anti-inflammatory, anti-inflammatory, anti-inflammatory, anti- (Park EK, et al., Int. Arch. Allergy Immunol., 133: 113-120, 2004).

The present invention relates to (1) a plant selected from the group consisting of at least one raw material selected from the group consisting of roots, stems, leaves and fruits of a plant of Panax species, and a plant selected from the group consisting of water and a lower alcohol having 1 to 4 carbon atoms Or more of a solvent; (2) The suspension prepared in the above step (1) is dissolved in a solvent selected from the group consisting of cellulase, amylase, pectinase, cellobiase and viscozyme. Preparing a mixed solution by mixing with the above-described saccharide-degrading enzyme; And (3) treating the mixed solution prepared in the step (2) at a temperature of 32 to 60 DEG C in an amount of 0.1 to 200 MPa to prepare an extract containing ginsenoside.

The method for producing ginseng extract comprises the steps of: (4) inactivating the glucosease contained in the extract prepared in the step (3); And (5) concentrating the extract in which the saccharolytic enzyme is inactivated in the step (4).

In step (1), the ginseng plant is selected from the group consisting of Panax ginseng, Panax quinquefolium, Panax notoginseng, Panax japonicum, Panax trifolium, Panax pseudoginseng, (Panax vietnamensis), and red ginseng, ginseng, white ginseng, cultivated ginseng, camellia ginseng, and wild ginseng depending on the processing condition and growth environment are also available.

In the step (1), the suspension contains 10 to 25 parts by weight, preferably 20 to 25 parts by weight, of at least one raw material selected from the group consisting of roots, stems, leaves and fruits of the plant of the genus Ginseng Mixed shows the best effect. When the concentration of the ginseng raw material in the solvent is out of the range of 10 to 25 parts by weight, the yield of the desired ginsenoside is lowered and the efficiency is lowered. Therefore, it is very important to mix the ginseng slurry with the solvent in the concentration range.

In addition, the suspension is preferably prepared by wet pulverizing roots, stems, leaves, fruits, or a mixture thereof of plants of the genus Ginseng with water, a lower alcohol such as methanol, ethanol, butanol, or a mixed solvent thereof.

In the step (2), the pH of the suspension is adjusted to 4.5 to 5, preferably 4.5 to 4.8 by adding a physiologically acceptable acid. pH is an important factor that directly affects the enzyme activity, and the type and yield of ginsenoside extracted depend on the small change of pH. In the present invention, at least one saccharide-degrading enzyme selected from the group consisting of cellulase, amylase, pectinase, viscose and cellobiase has a pH ranging from 4.5 to 5 , The yield of ginsenoside is lowered when it is out of the above range, and when the addition amount of the enzyme is increased to increase the yield of ginsenoside, economical efficiency and efficiency are inferior.

Wherein the physiologically acceptable acid is selected from the group consisting of inorganic acids including hydrochloric acid, sulfuric acid and phosphoric acid and organic acids including oxalic acid, acetic acid, succinic acid, tartaric acid, ascorbic acid, citric acid, glutamic acid, methanesulfonic acid and ethanesulfonic acid 1 or more.

In step (2), the saccharide-degrading enzyme is most preferably mixed with 1 to 5 parts by volume, preferably 2 to 4 parts by volume, more preferably 3 to 4 parts by volume of skin to 100 parts of the suspension.

The method of producing ginseng extract according to the present invention is characterized in that a method of extracting ginsenosides by adding a conventional enzyme to extract ginsenosides by applying pressure at an optimal pH and temperature, The enzyme of the present invention exhibits excellent ginsenoside yield and is highly economical.

In the step (3), the treatment temperature of the mixed solution is suitably 37 to 55 ° C, preferably 40 to 50 ° C, more preferably 40 to 45 ° C. The treatment temperature of various saccharide-degrading enzymes according to the present invention exhibits a synergistic effect of ginsenoside yield with optimal activity at an ultra-high pressure process is 37 to 55 ° C. When the above-mentioned range is exceeded, the enzyme activity is slowed and energy consumption is increased, And the extraction of the desired ginsenoside is not smooth.

In the step (3), the treatment pressure of the mixed solution is 0.1 to 150 MPa, preferably 50 to 150 MPa, and more preferably 50 to 100 MPa. As the treatment pressure of the mixed solution increases, the ginsenoside yield is high. However, since the ginsenoside extraction is not smooth when the treatment pressure is too low, the enzyme is added and the mixed solution is treated within the above range, .

At this time, sealing the mixed solution at the temperature and the pressure for 8 to 30 hours, preferably 12 to 24 hours, more preferably 18 to 24 hours, shows the optimum effect.

In the method of deactivating the glucosease in the step (4), the heat treatment is preferably performed at 70 to 100 ° C for 10 to 50 minutes, preferably at 70 to 90 ° C for 10 to 40 minutes.

The concentration in the step (5) is preferably lyophilized for 48 to 72 hours.

The ginseng extract is prepared by adding a mixed enzyme of pectinase and cellulase or a mixed enzyme of pectinase, cellulase, and cellulase to a glucosease digesting enzyme in step (2), and adding 95 to 105 MPa When treated at ultra high pressure, it shows a high ginsenoside content of 25 mg / g or more.

In the method for preparing ginseng extract according to the present invention, when cellulase is added together with other saccharide decomposing enzyme than when added alone, high ginsenoside content and minor ginsenoside can be obtained.

When the amylase, the cellulase, and the pectinase are added as the saccharide-degrading enzyme in the step (2), the content of ginsenosides Rg1 and Rb1 is greatly increased, and thus the ginseng extract can be used as a functional beverage composition. More specifically, it can be used as a functional beverage composition for anti-fatigue, antioxidation, immunity enhancement and the like. When the content of ginsenosides Rg1, Rb1 and Rg3 suggested by the food supplier is 2.5 to 34 mg / g, Fatigue improvement, antioxidation, and other functional health functional foods.

In this case, the amylase, cellulase and pectinase have a mixing ratio of 5: 20 to 0.5: 2 to 2 to 8, preferably 8 to 15: 1: 2 to 5, more preferably 10: ml) is the most effective. The amount of enzyme required to convert 1 μmol of substrate to product at 30 ° C / min under optimal conditions of enzyme is referred to as 1 unit, and the enzyme concentration in 1 ml is expressed as the unit of enzyme.

When the cellulase, the pectinase and the cellobiase are added as the carbohydrase in the step (2), the content of the ginsenoside Rd is greatly increased and the whitening, moisturizing, acne improvement, antioxidation, anti-aging, antibacterial , Antiinflammation, atopy, etc. (Kim et al., J Ginseng Res. (2013) 37: 54-63).

At this time, the mixing ratio of the cellulase, the pectinase and the cellobiase is 0.5 to 3: 0.5 to 3: 0.5 to 3, preferably 0.5 to 1.5: 0.5 to 2: 0.5 to 1.5, more preferably 1: (unit / ml), and it is preferable to adjust the pressure treatment time to 18 to 24 hours in order to extract a high content of ginsenoside Rd.

When the cellulase, amylase and cellobiase are added as the saccharide-degrading enzyme in step (2), the ginseng extract is mixed with ginsenoside Re, Rg1, Rc and Rb2 together with ginsenoside Rh1 and compound k , Which can be used as a pharmaceutical composition for anticancer therapy. With the extraction of major ginsenosides, it is possible to produce Rh1 and compound k in a simpler manner by converting major ginsenosides into minor ginsenosides.

At this time, the mixing ratio of the cellulase, the amylase, and the cellobiase is 0.5-3: 0.5-3: 0.5-3, preferably 0.5-1.5: 0.5-2: 0.5-1.5, more preferably 1: / ml) is the most effective.

The present invention provides ginseng extract extracted by the above-described method.

In addition, the present invention provides a health functional food for anti-fatigue, antioxidation, immunity enhancement and the like containing the ginseng extract as an active ingredient.

The health functional food may be prepared in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, solutions, aerosols, extracts, tea, jellies or drinks. More specifically, dairy products such as drink, meat, sausage, bread, biscuit, rice cake, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums and ice cream, various soups, beverages, alcoholic beverages and vitamins And includes all health foods in a conventional sense.

The ginseng extract according to the present invention can be added directly to the food or can be used together with other food or food ingredients, and can be suitably used according to conventional methods. The amount of the active ingredient to be mixed can be suitably determined according to the purpose of use (prevention or improvement). Generally, the amount of the extract in the health food may be 0.01 to 15% by weight of the total food, and the health beverage composition may be added in a proportion of 0.02 to 5 g, preferably 0.3 to 1 g, based on 100 ml have. However, the amount added in the case of long-term intake intended for health and hygiene purposes or for the purpose of controlling health may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range .

The health beverage composition according to the present invention is not particularly limited as long as it contains the above-mentioned extract at the indicated ratio as an essential ingredient, and may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages . Examples of natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides, such as maltose, sucrose, etc .; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol, erythritol and the like. As the flavoring agent, natural flavoring agents (tautatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavorings (saccharin, aspartame, etc.) can be advantageously used. The ratio is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

In addition to the above, the ginseng extract of the present invention can be used as a flavoring agent such as a variety of nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and thickening agents (cheese, chocolate etc.), pectic acid and its salts, Salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the ginseng extract of the present invention may contain flesh for the production of natural fruit juice, fruit juice drink and vegetable drink. These ingredients can be used independently or in combination, and the proportion of such additives is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the ginseng extract of the present invention.

The present invention also provides a composition for external application for skin comprising whitening, moisturizing, acne improvement, antioxidation, anti-aging, antibacterial, anti-inflammatory, atopic and the like containing the ginseng extract as an active ingredient.

The composition for external application for skin may be in the form of a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a surfactant-containing cleansing oil, a foundation or a spray. More specifically, the present invention relates to a composition for external application for skin comprising the extract of Ginseng of the present invention, such as a lotion, a skin lotion, a nutrition lotion, a nutritional cream, a massage cream, an essence, a pack, a cleansing, a cleanser, a soap, a treatment, Can be used in an amount of 0.001 to 90% by weight, preferably 0.01 to 10% by weight, based on the total weight of the composition.

The present invention also provides a pharmaceutical composition for antioxidation comprising a pharmaceutically effective amount of ginsenoside contained in the ginseng extract and a pharmaceutically acceptable carrier.

The present invention also provides a pharmaceutical composition for enhancing immunity comprising a pharmaceutically effective amount of ginsenoside contained in the ginseng extract and a pharmaceutically acceptable carrier.

Examples of carriers, excipients or diluents usable in the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose , Microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil.

The pharmaceutical composition may be in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, solutions, aerosols, excipients, injections, transdermal drugs or suppositories.

In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which are prepared by mixing at least one excipient, for example, starch calcium carbonate, sucrose or lactose, Prepare. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of liquid formulations for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweetening agents, have. 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, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.

The pharmaceutical composition may be administered parenterally or orally to consume ginseng extract at a daily dose of 0.0001 to 100 mg / kg, preferably 25 to 100 mg / kg, more preferably 50 to 100 mg / kg. The dosage may be administered once a day or divided into several doses. The dose may be varied depending on the progress of the disease, age, sex, physical condition, administration period, administration method, body weight of the patient, ≪ / RTI >

The method of producing ginseng extract according to the present invention as described above comprises not only a high amount of major ginsenosides but also minor ginsenosides, so that the ratio of Rg1 and Rb1 and the ratio of Re, Rc, Rb2, Rg2, Rf, Rd, k (compound K) is high.

In addition, the active material can be easily extracted through a simple manufacturing process, and can be variously applied to a health functional food, a skin external composition and a pharmaceutical composition.

Various formulation examples and formulations including the ginseng extract of the present invention are described below, but the present invention is not intended to be limited thereto but is specifically described.

Preparation Example 1. Preparation of tablets

Example Ginseng Extract 100 mg

Starch ............................................ 50mg

Lactose ............................................. 50mg

Magnesium stearate ............................. 5mg

The above components were mixed and tablets were prepared by a wet granulation method or a dry granulation method according to a conventional method for producing tablets.

Formulation Example 2. Preparation of capsules

Example Ginseng Extract 100 mg

Starch ................................................. ...... 50 mg

Lactose ................................................. ...... 50 mg

Talc ................................................. ...... 1 mg

Magnesium stearate ....................................... 5mg

The above components were mixed and capsules were prepared according to the conventional preparation method of capsules.

Formulation Example 3. Preparation of a liquid preparation

Example Ginseng Extract 500 mg

Isolation Party ................................................ ..10g

saccharose................................................. ...... 10 g

Lemon flavor ................................................ ..... Appropriate amount

Purified water was added to the whole ....................................... 50 ml

The above components were mixed according to a conventional method for preparing a liquid preparation, filled in a brown bottle, and sterilized to prepare a liquid preparation.

Formulation Example 4. Preparation of injection

Example Ginseng Extract .................................... 50 mg

Sodium Metabisulfite ................................... 2.6mg

Methylparaben .............................................. 0.5 mg

Propylparaben ............................................ 0.1㎎

Sterilized distilled water for injection ........................................ Appropriate amount

The above ingredients were mixed and adjusted to 2 ml in a conventional manner, filled in an ampoule and sterilized to prepare an injection.

Formulation Example 5. Preparation of beverage

Example Ginseng Extract ...................................... 100 mg

Vitamin B2 ................................................ .... 5 mg

Vitamin B6 ................................................ .... 5 mg

Vitamin C ................................................ .... 80 mg

Dietary Fiber................................................ .1,000 mg

Liquid fructose ................................................ 10,000 mg

Emulsifier ................................................. .... 100 mg

Lemon flavor ................................................ ...... 50 mg

Purified water................................................. ... Appropriate amount

The above components were mixed to make a volume of 50 ml, the mixture was passed through a 3-μm filter, and the resulting filtered mixture was first sterilized at 90 to 93 ° C for 20 seconds, filled in a 50 ml bottle, Followed by secondary sterilization for 20 minutes to prepare a beverage product.

Formulation Example 1. Flexible lotion (skin lotion)

Example Ginseng Extract 2.0

Glycerin ............................... 3.5

Oleic alcohol ........................... 1.5

Ethanol ................................. 5.5

Polysorbate 80 ....................... 3.2

Carboxyl Vinyl Polymers ..................... 1.0

Butylene glycol .......................... 2.0

Propylene glycol ........................ 2.0

Preservative, fragrance ............................

Purified water ................................. Remaining amount

Total ...................................... 100

Flexible lotion was prepared according to the above composition in a conventional manner (unit weight%).

Formulation Example 2. Nourishing lotion (milk lotion)

Example Ginseng Extract 2.0

Glycerin ...................................... 3.0

Butylene glycol .................................. 3.0

Propylene glycol ................................ 3.0

Carboxyvinyl polymer .............................. 0.1

Wax ........................................... 4.0

Polysorbate 60 ............................... 1.5

Caprylic / capriciculiglyceride .................. 5.0

Squalane ....................................... 5.0

Solvatescequioleate ........................... 1.5

Cetearyl Alcohol ................................. 1.0

Triethanolamine ................................. 0.2

Preservative, fragrance ....................................

Purified water ........................................... Remainder

Total ............................................... 100

Nutrition lotion was prepared according to the above composition in a conventional manner (unit weight%).

Formulation Example 3. Nourishing cream

Example Ginseng Extract 2.0

Glycerin ....................................... 3.5

Butylene glycol .......................................

Liquid paraffin ..................................... 7.0

Beta Glucan ..................................... 7.0

Carbomer .......................................... 0.1

Caprylic / capric wriglyceride .................. 3.0

Squalane ........................................ 5.0

Cetearyl Glucoside ........................... 1.5

Sorbitan stearate ............................ 0.4

Polysorbate 60 ................................. 1.2

Triethanolamine ................................... 0.1

Preservatives, fragrances .....................................

Purified water ........................................... Remainder

System ................................................ 100

Nutritive creams were prepared according to the above composition in a conventional manner (unit weight%).

Formulation Example 4. Pack

Ginseng Extract of the Example ... 2.0

Glycerin ..................................... 4.0

Polyvinyl alcohol ................................. 15.0

Hyaluronic Acid Extract 5.0

Beta Glucan ................................... 7.0

Allantoin ...................................... 0.1

Nonyl phenyl ether ............................... 0.4

Polysorbate 60 ............................... 1.2

Preservative, fragrance ..................................

Purified water ........................................ Residue

Total ............................................. 100

A pack was prepared according to the above composition in a conventional manner (unit weight%).

Formulation Example 5. Pharmaceutical preparation for local administration (patch)

Example Ginseng Extract 2.0

Beta-1,3-glucan ........................................... ... 3.0

Diethylamine ............................................... .... 0.7

Sodium Sulfite ................................................ .0.1

Polyoxyethylene lauryl ether (EO = 9) ............................ 1.0

Polyhydroxyethylene cetyl stearyl ether (Cetomacrogol 1000) ... 1.0

Viscous paraffin oil ............................................ 2.5

Caprylic acid ester / capric acid ester (Cetiol LC) 2.5

Polyethylene glycol 400 ........................................ 3.0

Polyacrylic acid (Carbopol 934P) ................................... 1.0

Purified water................................................. ....... balance

system................................................. ........... 100

According to the above composition, a medicament for topical administration was prepared by a conventional method (unit% by weight).

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 embodiments are merely illustrative of the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

Examples. Manufacture of ginseng extract

In the examples of the present invention, commercially available enzyme cellulases (from Trichoderma reesei ATCC 26921 - Sigma C2730) listed in Table 1, α-amylase (from Bacillus licheniformis - Sigma A3403), pectinase (from Aspergillus aculeatus - Sigma P2611), viscose L (Sigma V2010) and cellobiase (from Aspergillus niger - Sigma C6105) and 4 year old, 6 year old ginseng ginseng and 4 year old ginseng ginseng were used.

400 g of 4-year-old ginseng roots were finely chopped and homogenized with 2 L of sterilized distilled water using a mixing grinder. The pH of the suspension was adjusted to 4.0 to 4.8 with citric acid. Thereafter, as shown in Table 2, enzymes were combined, and 2 to 4 parts of the skin enzyme was mixed with 100 parts of the suspension in the pH adjusted suspension, and the mixture was filled in a plastic container having a flexible wall and sealed, 25, 50, 75 and 100 MPa at 45 ° C for 24 hours with high pressure or high hydrostatic pressure (HHP) treatment (HHP machine, TFS-2L, Toyo-Koatsu Innoway Co. Ltd., Hiroshima, Japan) To obtain an extract.

Controls without enzyme treatment were also compared in HHP and AP.

Thereafter, the extract was treated at 80 캜 for 30 minutes, cooled in an ice bath, compressed by a sieve of 250 탆, lyophilized for 48 to 72 hours, concentrated, -20 < [deg.] ≫ C to carry out the following experiment.

Experimental Example 1. Measurement of total saponin (ginsenoside) content

The content of total ginsenosides extracted was determined by the method of Wei, SE, 2011 (Isolation and determination of anti-nutritional compounds from root and shells of Peanut ( Arachis hypogaea ), Department of Chemical Science, Faculty of Science, Universiti Tunku Abdul Rahman, Selangor, Malaysia, p. 104.).

A methanol aqueous solution having a concentration of 80% was added to each sample (100 μl) to adjust the volume to 0.25 ml. Then, 0.25 ml of a vanillin reagent having a concentration of 8% and 2.5 ml of sulfuric acid having a concentration of 72% were added and mixed. Thereafter, the sample was treated in a water bath at 60 ° C for 10 minutes, cooled at room temperature, and absorbance was measured at 544 nm. The standard curve was obtained using various concentrations of diosgenin as a standard saponin, 80% methanol aqueous solution for standard saponin solution, and reagent blank for 80% aqueous methanol solution instead of sample containing extract 0.25 ml was used.

Experimental Example 2. HPLC analysis of ginseng saponin (ginsenoside)

The ginseng extract extracted from HHP and AP was shaken with n-butanol having the same volume at room temperature for 20 minutes and centrifuged at 13,000 g for 5 minutes for phase separation. The butanol layer was collected and vacuum evaporator ≪ / RTI > and then dissolved in methanol for HPLC analysis.

HPLC analysis was carried out on a Waters HPLC system using a 4.5 mm × 25 cm Sunfire C18 column, using HPLC acetonitrile and water as mobile phase and at a flow rate of 1 ml / min. In addition, 20 μl of each sample was filtered through a 0.25 μm PTFE filter before being injected into the C18 column.

The elution of the various ginsenosides was observed at 203 nm and the order and amount of elution of ginsenoside was calculated by comparison with the chromatogram of standard ginsenoside mixture (Cerillant co., USA). HPLC analysis for each sample was repeated three times for accuracy.

Experiment result.

(1) Results of extraction of ginsenoside with changes in pressure.

The contents of ginsenosides extracted under various pressures in the above examples are shown in Fig.

As a result, when the extraction was performed at 0.1 to 100 MPa, the content of ginsenoside was 12 to 24 mg / g and the content of ginsenoside was 18 to 24 mg / g when extracted at 50 to 100 MPa, The content of ginsenosides was increased and showed maximum at 100 MPa.

(2) Ginsenoside Extraction Result According to Different Ginseng Types.

The contents of total ginsenosides were compared by treating 4-year-old Geumsan ginseng, 6-year old Geumsan ginseng and 4-year old ginseng ginseng at normal pressure (0.1 MPa) and ultrahigh pressure (100 MPa), respectively.

As a result, as shown in FIG. 3, 4-year-old Geumsan ginseng treated with 100 MPa of super high pressure showed the highest total ginsenoside content, and HPLC analysis showed that 4-year old Geumsan ginseng had the highest major ginsenoside ginsenoside content.

(3) Extraction results of ginsenosides according to the change of ginseng concentration on solvent.

In order to examine the concentration of the ginseng slurry mixed in the solvent for extracting a high content of ginsenoside, ginseng roots 2, 5, 10, 15, 20 and 25 parts by weight were mixed with 100 parts by weight of distilled water by the method of the above- And 100 MPa, respectively, to prepare ginseng extract.

As a result, as shown in FIG. 4, when 20 parts by weight of ginseng was added to 100 parts by weight of distilled water, the total ginsenoside content extracted was the highest.

(4) Results of extraction of ginsenosides with various combinations of enzymes.

The total ginsenoside content extracted at atmospheric pressure (0.1 MPa) and ultrahigh pressure (100 MPa) according to the combination of various enzymes by the method of the above Example is shown in FIG.

As a result, it was shown that when pectinase and cellulase or pectinase, cellulase and cellobase were added, they had a high ginsenoside content of 25 mg / g or more.

(5) Results of ginsenoside extraction according to the pH of the suspension.

To determine the optimum pH for the added cell wall degrading enzyme reaction, the pH of the suspension homogenized by mixing the ginseng slurry (ginseng roots) with the solvent in the above example was changed (pH 4, 4.5, 4.8 , Adjusted to 5, adjusted to pH 5.5, 6, 6.5, 7 with 2 M trisodium citrate), and the yield of ginsenosides extracted at 45 ° C (100 MPa) at 12 h was compared. The yield of ginsenoside was the highest at 4.8 (see FIG. 6).

(6) Results of extraction of ginsenoside with pressure treatment temperature.

The yield of ginsenosides extracted at an ultra-high pressure (100 MPa) was examined by varying the ultrahigh-pressure treatment temperature at 25 to 60 ° C by the method of the above-mentioned Example. As a result, the treatment was carried out at 45 ° C at 100 MPa The yield of ginsenoside was the best (see Fig. 7).

(7) Results of ginsenoside extraction with pressure treatment time.

8 shows the ginsenosides contained in the extract prepared by treating the ginseng extract by the method of the above example at 12 atm and 100 MPa at normal pressure (100 MPa) for 12 hours and 24 hours, respectively.

(8) Results of addition of amylase, cellulase and pectinase as digestive enzymes.

(5 units / ml), cellulase (0.5 Unit / ml) and pectinase (2 units / ml) were mixed with the enzyme to prepare an extract of ginseng at an atmospheric pressure (0.1 MPa) MPa), it was found that Rg1 and Rb1 were extracted at the same ratio as shown in FIG. 9, and the yield was greatly increased.

(9) The result of addition of cellulase, pectinase and cellobiase as a digestive enzyme.

(2 units / ml), pectinase (2 units / ml) and cellobiase (2 units / ml) were added to the extracts for 12, 18 and 24 hours (0.1 MPa) and ultrahigh pressure (100 MPa), respectively.

As a result, Rg1, Re, Rc, Rb2 and Rb1 were extracted together with Rd. As shown in FIG. 10, the Rd content was greatly increased upon extraction for 18 and 24 hours, and the HdP- (Rd) content was 1.73 times higher than that obtained at atmospheric pressure (AP-CPC), which was 2.57 ± 0.128 mg / g.

(10) Results of addition of cellulase, amylase, and cellulase with the digestive enzymes.

Cellulase (2 Unit / ml), amylase (2 Unit / ml) and cellobiase (2 Unit / ml) were added as an enzyme to the ginseng extract by the method of the above example, (0.1 MPa) and ultrahigh pressure (100 MPa).

As a result, Rg1, Re, Rc, Rb2 and Rb1 were extracted together with Rh1 and compound k, and the highest minor dienes Rh1 and compound k contents were obtained when they were extracted at high pressure (HHP-CAC) for 24 hours. According to the method for preparing ginseng extract according to the present invention, when the cellulase, amylase and cellobiase are added as an enzyme and the treatment with ultrahigh pressure is performed, the major ginsenosides are extracted and major ginsenosides are converted into minor ginsenosides, And compound k (see Table 3). Table 3 shows the results of extraction at normal pressure (0.1 MPa) and ultrahigh pressure (100 MPa) for 24 hours.

(11) Results of ginsenoside extraction by inactivation of the digestive enzymes.

In the above example, the temperature and time for inactivating the enzyme after 100 MPa of ultra-high pressure treatment were treated at 80 캜 for 30 minutes, 100 캜 for 30 minutes, and 121 캜 for 15 minutes to prepare respective ginseng extracts.

As a result, when the extract was treated at 80 ° C for 30 minutes, ginsenosides could be extracted without changing the concentration of ginsenosides occurring during inactivation of the enzyme.

Having described specific portions of the present invention in detail, it will be apparent to those skilled in the art that this specific description is only a preferred embodiment and that the scope of the present invention is not limited thereby. It will be obvious. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (21)

delete delete delete delete delete delete delete delete delete delete delete delete delete A method for producing an extract containing ginsenoside useful components ginsenoside Rh1 and compound k (compound k)
(1) Panax species: at least one solvent selected from the group consisting of roots, stems, leaves and fruits of plants, and at least one solvent selected from the group consisting of water and lower alcohols having 1 to 4 carbon atoms; To obtain a suspension;
(2) mixing the suspension prepared in the step (1) with a mixed enzyme consisting of cellulase, amylase and cellobiase in a ratio (unit / ml) of 0.5-3: 0.5-3: 0.5-3 ; And
(3) treating the mixed solution prepared in the step (2) at a temperature of from 32 to 60 DEG C with 0.1 to 200 MPa to prepare an extract containing ginsenosides, wherein the ginsenosides useful components ginsenosides Rh1 and k (compound k).
delete 15. The method of claim 14,
A method for producing an extract containing a useful component ginsenoside Rh-1 and a compound (k), which is a useful component of ginseng, is characterized in that the ginsenoside-containing component ginsenoside Rh1 and the compound k-containing extract are used as an anticancer pharmaceutical composition.
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