JPS62158490A - Treatment of panax ginseng - Google Patents

Abstract

PURPOSE:To obtain Panax ginseng having high ginsenoside Rh content and preservable over a long period, at a low cost, by treating a tissue culture product of Panax ginseng at a high temperature and drying under specific condition. CONSTITUTION:A tissue culture product of Panax ginseng is heat-treated in a closed vessel at 110-160 deg.C for 0.5-24hr. The ginsenoside Rh content of the tissue is increased by this process. The heat-treated product is dried at a temperature lower than the above heat-treatment temperature, i.e. with hot air of 40-60 deg.C to enable the long-term preservation of the tissue.

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial Application Field) The present invention is directed to the use of ginsenoside Rh, which is said to have an anticancer effect.
The present invention relates to a method for processing medicinal ginseng to increase the content of medicinal ginseng, particularly a method for processing a tissue culture of medicinal ginseng.

(Prior art) Medicinal ginseng 2 For example, Panax gins
engC18, Meyer), Chikusetsu ginseng (r'a
nax japonicus C, A, Meyer),
American ginseng (Panax quinquefol)
), Panax noLo(Hi
nseng (Burk) F.

If, Chen), Siberian ginseng (Eleut)
herococcus senticosus) root is prized and widely used as a useful herbal medicine. Its medicinal properties have long been said to include tonicity and longevity, and its sedative, stimulant, and diuretic effects have now been revealed. The active ingredients of medicinal ginseng are mainly saponins. The saponins of medicinal ginseng are collectively called ginsenosides, and to date they have been classified as ginsenoside Ro, ginsenoside Ra, ginsenoside Rb, ginsenoside Rc, and ginsenoside Rd.

Ginsenoside Re, ginsenoside Rf, ginsenoside R,g and ginsenoside Rh are known.

It has been reported that ginsenoside Rh has a sedative effect and ginsenoside Rg has a stimulatory effect (
Pharmaceutical Journal Vol. 82, No. 12, pp. 1633-1634; Protein.

Nucleic Acids, Enzymes, Vol. 12, No. 1, pp. 32-38). Ginsenosides 11c, Rd, and Re have also been reported to have physiological activities such as promotion of serum cholesterol synthesis.

Ginsenoside Rh is said to have anticancer effects, but
The ginsenoside Rh content of natural medicinal ginseng is extremely low and may not be detected in some cases.

That research is lagging behind. If ginseng containing a large amount of ginsenoside Rh can be obtained, research on ginsenoside Rh will progress and its medicinal efficacy will be clarified.

(Problems to be Solved by the Invention) The present invention solves the above-mentioned conventional drawbacks, and its purpose is to provide a method for increasing the content of ginsenoside Rh in medicinal ginseng. Another object of the present invention is to utilize a ginseng tissue culture that is easier to prepare than natural ginseng.

It is an object of the present invention to provide a treatment method for increasing the amount of ginsenoside Rh contained in this.

(Means and effects for solving the problems) The method for treating medicinal ginseng of the present invention includes the step of treating a tissue culture obtained from medicinal ginseng at high temperature to increase the content of ginsenoside Rh, thereby increasing the content of ginsenoside Rh. The purpose is achieved.

The medicinal ginseng tissue culture used in the present invention is:

Obtained by conventional plant tissue culture methods. for example.

Panax ginseng, Panax ginseng, American ginseng, Panax ginseng,
Tissues of known medicinal ginseng, such as Siberian ginseng, are cultured to generate callus. Next, the obtained callus is grown by a liquid culture method or a solid culture method. This callus can be grown indefinitely by culturing. Culture conditions do not need to be particularly special. The medium used is Murashige-5koog, which is commonly used for plant culture.
g) medium, White's medium, O, L, Gamborg (0,L).

Gamborg's B5 medium, Nitsch
) medium.

Heller's medium, Morel
It is possible to use a medium such as 1). If necessary, casein degrading enzyme, soybean flour, cornstarch liquor, vitamins, etc. can be added to this.

Details of the culture method can be found in 1, for example, JP-A-59-169486, JP-A-59-169487, JP-A-59-
It is disclosed in Japanese Patent No. 16488. If liquid culture method is adopted.

The resulting tissue culture cell mass does not become extremely large when shaken or stirred, and its maximum diameter is approximately 15 mm at most. Since such cell clusters have uniform particle sizes, they are convenient for extracting ginsenosides after high-temperature treatment as described below. The obtained cell culture is subjected to a ninth high-temperature treatment after removing excess water by filtration, centrifugation, etc., if necessary.

The high temperature treatment is carried out by heating the above tissue culture at a temperature of 110 to 160%.
C.1, preferably 120 to 150.degree.

The treatment time is usually 0.5 to 24 hours, preferably 1 to 1
It is 0 hours. For example, at 110℃, the high temperature treatment time is 5
It takes about an hour. If the processing temperature is high, shorten the processing time. If the treatment temperature is lower or the treatment time is shorter than the above range, the ginsenoside Rh content of the resulting ginseng will be low. On the other hand, if the processing temperature is too high or the processing time is too long, there is a risk that ginsenosides and other active ingredients will decompose due to heat.
Ru. The tissue itself also turns brown.

When the high temperature treatment is performed in this manner, the cell tissue of the 1Mi textile culture is destroyed and the cell fluid is concentrated. Since cell fluid inherently contains organic acids, concentrated cell fluid is placed under strongly acidic conditions. It is thought that under such acidic conditions, the ginsenosides contained become more susceptible to hydrolysis. for example.

Ginsenosides Re and Rg are compounds in the form of protopanaxytriol bound to a molecule such as glucose, and when this is hydrolyzed, the sugar moiety is eliminated and ginsenoside Rh is produced. Especially ginsenoside R [
Since the Th-ffi content in medicinal ginseng is relatively high, ginseng containing a large amount of ginsenoside Rh can be obtained by performing high temperature treatment. The high temperature treatment is preferably carried out in a sealed container. When treated in a sealed container, a tissue culture is treated at high temperature under high humidity, and under such conditions, the hydrolysis reaction of ginsenoside is promoted. The obtained ginseng contains ginsenoside Rh at a rate about five times that of natural medicinal ginseng.

The moisture content of carrots after high-temperature treatment varies somewhat depending on the conditions of the high-temperature treatment, but is usually 50 to 80%. If this is to be stored, it is then subjected to a drying process. The drying temperature is lower than the above-mentioned high temperature heat treatment temperature, and is usually 40 to 60°C. Ventilation drying (warm air drying) is preferred. The time required for drying is usually 1 hour or more, preferably 6 to 24 hours. If the drying temperature is too high, active ingredients such as ginsenosides will decompose, and if the drying temperature is too low, it will take a long time to dry, which may lead to the growth of mold and deteriorate quality.

(Example) The present invention will be described below with reference to Examples.

Tenkawa 1 to 21 (A) Cultivation of medicinal ginseng: A part of the tissue of medicinal ginseng (Panese ginseng) was cut out and treated with 0.5 each of 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin.
Tissue culture was carried out for 1 to 2 months in MS medium containing ppm. The obtained callus was treated with 2 ppm of indole acetic acid (IAA).
and kinetin 0. The cells were cultured for one month using a modified MS medium containing lppm. Approximately 500 g of tissue culture with a moisture content of 94% was obtained.

(B) High-temperature treatment of carrots: The tissue culture obtained in section (^) was placed in a sealed container with a capacity of 3000 cal and subjected to high-temperature treatment at 150° C. for 5 hours.

(C) Extraction of saponin = 200ml of 50% ethanol was added as an extraction solvent to 5g of the high-temperature-treated carrots obtained in section (B), and the mixture was immersed at 50°C for 24 hours to perform extraction.

The extract obtained by distilling off the extraction solvent was dissolved in 100 ml of distilled water, 50 mj2 of diethyl ether was added, and the mixture was shaken and separated using a separatory funnel to obtain an aqueous layer, and the diethyl ether soluble portion was removed. Separately, n-butanol and 1% NaCl aqueous solution (using distilled water) were mixed at a ratio of 7:3 to prepare water-saturated butanol.

100 m/ml of this water-saturated butanol was added to the aqueous layer, shaken and separated, and saponin was extracted into the butanol layer. In the water layer, there is also 50m7 of water-saturated butanol! Add and shake.
Separate and discard the aqueous layer to obtain a butanol layer. After combining this butanol layer and the above-mentioned butanol layer, washing was performed using 100+ylN of butanol saturated water. The butanol saturated water used here was prepared by mixing a 1% NaC] aqueous solution (using distilled water) and n-butanol at a ratio of 7:3. After washing, 50 ml of butanol saturated water was further added to the butanol layer for washing. After combining the aqueous layers obtained by washing, 50 mff of water-saturated butanol was added to recover saponin from the aqueous layer. Combine the obtained butanol layer and the above washed butanol layer in an evaporator? He worked on agriculture and obtained crude saponin.

(D) Detection of ginsenosides: The crude saponin obtained in section (C) was dissolved in 10 ml of methanol. Place this at a distance of 5 μm from the longitudinal end of the thin layer plate.
2 spots, water-ethyl acetate-n-butanol (5:
1:4) 19.5c as the developing solvent for the upper layer of the mixture
m expanded. After the developed thin layer plate was air-dried for 24 hours, a 10% aqueous sulfuric acid solution was uniformly sprayed thereon. After heating this plate in a dryer at 120°C for 7 minutes, using a high-speed thin layer chromatography scanner C5-920 (manufactured by Shimadzu Corporation),
Measurements were taken at 530 nm. The obtained chart is shown in FIG. In FIG. 1, 1 represents the crude saponin obtained in this example, and 2 represents a standard sample containing ginsenoside Rh and Rg+.

1 peak O+ a+ b+ C1d+ e+
f+ g and h are ginsenoside Ro, I, respectively
la, Rh, Rc.

Rd, Re, Rf and Rg are shown. In 2, peak b1 and peak gl are respectively ginsenoside R.
b, and ginsenoside Rgl are shown. Table 1 shows the peak position and ginsenoside content of each ginsenoside. In Table 1 and Tables 2 to 4 described below, the peak position indicates the distance from the plate edge of the thin layer plate to each ginsenoside spot.

Crude saponin) 8 solution was spotted at a distance of 201 from the edge of the plate, so the value obtained by subtracting 20 from the value shown in Table 1 corresponds to the spread distance. Ginsenoside content fffi is a relative value indicating the area of each peak.

Table 1 (^) Culture of medicinal ginseng: Example 1-1 Same as section (A).

(B) Two high-temperature treatments of carrots M obtained in Example (A)
The i-woven culture was treated at high temperature in the same manner as in Example 1-1 (It), then laminated to a thickness of about 1 am and heated at 50°C for 3.
Dry with warm air for 24 hours at an air volume of 5%/n+in, 36g
A dried product was obtained.

(C) Extraction of saponin 2. 5 g of dried ginseng obtained in Example (B) was treated according to Example 1-1 (C) to obtain crude saponin.

(D) Detection of ginsenosides 2.Example 1-1
Detection was performed according to section (D), and a chart almost identical to that in FIG. 1 was obtained.

Large rattan charcoal main two (A) Cultivation of medicinal ginseng: Example 1-1 Same as section (A).

(B) High-temperature treatment of carrots (2) The tissue culture obtained in Example (A) was placed in a sealed container with a capacity of 3000 cnl.
High temperature treatment was performed at 0°C for 5 hours.

(C) Extraction of saponin 2 Using 5 g of the high-temperature-treated ginseng obtained in Example (Il), the treatment was performed according to Example 1-1 (C) to obtain crude saponin.

(D) Detection of ginsenosides 2.Example 1-1
Detection was performed according to section (D), and the chart shown in FIG. 2 was obtained. Table 2 shows the peak position and ginsenoside content of each ginsenoside.

Table 2 (8) Cultivation of medicinal ginseng: Same as Example 1-1 (A).

(13) High-temperature treatment of carrots (2) The tissue culture obtained in Example (8) was treated at high temperature in the same manner as in Example 2-1 (B), and then layered to a thickness of about 1 cm. 3. At 60°C.
5n? /min, hot air drying for 24 hours, 35g
A dried product was obtained.

(C) Extraction of saponin 2. 5 g of dried ginseng obtained in Example (B) was treated according to Example 1-1 (C) to obtain crude saponin.

(D) Detection of ginsenosides: Ginsenosides in the crude saponin obtained in Section (C) of this Example were detected in Example 1-1 (
Detection was performed according to Section D), and a chart almost identical to that shown in Figure 2 was obtained. The results of Example 2-2 are shown in Figure 5. In Figure 5, the ginsenoside content is expressed in mg per gram of ginseng after high temperature treatment and drying.

(A) Culture of medicinal ginseng: Same as Example 1-1 (A).

(B) High-temperature treatment of carrots: The tissue culture obtained in section (A) of this example was placed in a sealed container with a capacity of 3000 cm, and heated for 13 minutes.
High temperature treatment was performed at 0°C for 5 hours.

(C) Extraction of saponin: Using 5 g of the high-temperature-treated ginseng obtained in Section (B) of this Example, the treatment was performed in accordance with Section (C) of Example 1-1 to obtain crude saponin.

(D) Detection of ginsenosides: Ginsenosides in the crude saponin obtained in Section (C) of this Example were detected in Example 1-1 (D
), the chart shown in FIG. 3 was obtained. Table 3 shows the peak position and ginsenoside content of each ginsenoside.

Table 3 Sl side LL2I (8) Culture of medicinal ginseng: Example 1-1 Same as section (A).

(B) High-temperature treatment of carrots: The tissue culture obtained in Section (8) of this Example was treated at high temperature in the same manner as in Example 3-1 (B), and then laminated to a thickness of about 2 CI11 at 60°C. 3.5r at
It was dried with hot air for 24 hours at an air flow rate of rr/min to obtain 35 g of a dried product.

(C) Extraction of saponin 2. 5 g of dried carrots obtained in Example (B) was treated as in Example 1-1 (C) to obtain crude saponin.

(D) Detection of ginsenosides Two ginsenosides in the crude saponin obtained in Example (C) were detected in Example 1-1 (
Detection was performed according to section D), and a chart almost identical to that shown in FIG. 3 was obtained.

(A) Culture of medicinal ginseng: Example 1-1 Same as section (A).

(B) High-temperature treatment of carrots (2) The tissue culture obtained in Example (8) was treated at high temperature in the same manner as in Example 1-1 (B), and then laminated to a thickness of about 2 cffl at 60°C. 3.5m at
The product was dried with hot air for 24 hours at an air flow rate of /min to obtain 35 g of dried product.

(C) Extraction of saponin: 5 g of dried ginseng obtained in section (B) of this example was treated according to section (C) of Example 1-1 to obtain crude saponin.

(D) Detection of ginsenosides Ginsenosides in the crude saponin obtained in Example (C) were detected in Example 1-1 (
Detection was performed according to section D), and a chart almost identical to that shown in FIG. 1 was obtained.

Comparative Example 1 (8) Cultivation of medicinal ginseng: Same as Example L-1 (A).

(B) Extraction of saponin: The tissue culture obtained in section (A) of this comparative example was layered to a thickness of about 2 cm and heated at 60°C for 3.
Hot air drying was performed for 24 hours at an air flow rate of 5 m/min to obtain 38 g of dried product. Using 5 g of this dried product, Example 1-1
Crude saponin was obtained by processing according to section (C).

(C) Detection of ginsenosides Two comparative examples Ginsenosides in the crude saponin obtained in section (B) were detected in Example 1-1 (
Detection according to section D) confirmed that almost no ginsenoside Rh was contained.

(8) Cultivation of medicinal ginseng: Same as in Example 1-1 (A).

(B) Two high-temperature treatments of carrots The tissue culture obtained in Example (^) was placed in a sealed container with a capacity of 3000 c-, and
High temperature treatment was performed at ℃ for 5 hours.

(C) Extraction of saponin 2 Using 5 g of the high-temperature-treated ginseng obtained in Example (B), the treatment was performed according to Example 1-1 (C) to obtain crude saponin.

(D) Detection of ginsenosides: Ginsenosides in the crude saponin obtained in section (C) of this comparative example were detected in Example 1-1 (
Detection was performed according to section D), and the chart shown in FIG. 4 was obtained. In FIG. 4, 3 indicates the crude saponin obtained in this comparative example, and 4 indicates a standard sample containing ginsenosides Rh and Rg.

Table 4 shows the peak position and ginsenoside content of each ginsenoside.

Table 4 (8) Cultivation of medicinal ginseng: Same as Example 1-1 (A).

(B) High-temperature treatment of carrots Two The tissue culture obtained in Example (^) was treated at high temperature in the same manner as in Comparative Example 1-1 (B), then laminated to a thickness of about 2 cm and heated to 60°C. 3゜5rrr at
The product was dried with hot air for 24 hours at an air flow rate of /min to obtain 37 g of a dried product.

(C) Extraction of saponin: 5 g of dried ginseng obtained in section (B) of this comparative example was treated according to section (C) of Example 1-1 to obtain crude saponin.

(D) Detection of ginsenosides Two comparative examples Ginsenosides in the crude saponin obtained in section (C) were detected in Example 1-1 (D
), a chart almost identical to that in FIG. 4 was obtained.

From Tables 1 to 4 and Figures 1 to 5, it is clear that ginseng containing a large amount of ginsenoside Rh can be obtained by high temperature treatment.

(Effects of the Invention) According to the present invention, ginseng containing a large amount of ginsenoside Rh can be obtained by treating a medicinal ginseng cell culture at high temperature. By drying this carrot under specific conditions, it can be made into dried carrot without any change in quality and can be stored for a long period of time. Since the medicinal ginseng cell culture used can be obtained in large quantities in a short period of time by conventional tissue culture methods, ginseng containing a large amount of ginsenoside Rh can be obtained at low cost. It is expected that research on ginsenoside Rh can be further advanced using such ginseng.

4. Figures 1 to 3 are thin-layer chromatograph charts showing each ginsenoside in saponin extracted from medicinal ginseng obtained by the method of the present invention; Figure 4 is a chart obtained by the method of the present invention A chart taken using a thin layer chromatography scanner showing the ginsenosides in saponin extracted from medicinal ginseng treated at a lower temperature; and Figure 5 compares the content of each ginsenoside in the ginseng obtained when the high temperature treatment temperature is changed. This is a graph.

that's all

Claims (1)

[Scope of Claims] 1. A method for treating medicinal ginseng, which includes the step of treating a tissue culture obtained from medicinal ginseng at high temperature to increase the content of ginsenoside Rh. 2. The treatment method according to claim 1, wherein the high temperature treatment is performed in a sealed container. 3. The treatment method according to claim 1, wherein the high temperature treatment is performed at 110 to 160°C for 0.5 to 24 hours. 4. A method for treating medicinal ginseng, which includes the steps of treating a tissue culture obtained from medicinal ginseng at a high temperature to increase the content of ginsenoside Rh, and then drying it at a lower temperature than the high temperature treatment temperature. 5. The treatment method according to claim 4, wherein the high temperature treatment is performed in a sealed container. 6. The treatment method according to claim 4, wherein the high temperature treatment is performed at 110 to 160°C for 0.5 to 24 hours. 7. The processing method according to claim 4, wherein the drying process is hot air drying at 40 to 60°C.