WO2011059265A2

WO2011059265A2 - Process for preparing extract fraction enriched with ginsenoside rg3 from ginseng radix

Info

Publication number: WO2011059265A2
Application number: PCT/KR2010/008004
Authority: WO
Inventors: Jin Hee Lee; Seok Jun Park; Yong Ki Seo; Yeong Eun Kim; Min Ju Park
Original assignee: Cj Cheiljedang Corp.
Priority date: 2009-11-13
Filing date: 2010-11-12
Publication date: 2011-05-19
Also published as: KR20110052940A; WO2011059265A3

Abstract

A method of preparing a ginseng radix extract fraction enriched with ginsenoside Rg3, the method including: enriching with ginsenoside Rg3 a ginsenoside Rb1-containing Ginseng Radix ethanol extract by reacting the ethanol extract with a mixture of water and acetic acid; adsorbing the ethanol extract enriched with ginsenoside Rg3 to an adsorption resin; allowing distilled water to go through the adsorption resin to elute components that are not adsorbed; adding 30 to 50 v/v% ethanol to the adsorption resin to remove non-saponin components and triol ginsenosides except for ginsenoside Rg3; and adding 50 to 100 v/v% ethanol to the adsorption resin to obtain an eluate.

Description

PROCESS FOR PREPARING EXTRACT FRACTION ENRICHED WITH GINSENOSIDE RG3 FROM GINSENG RADIX

This application claims the benefit of Korean Patent Application No. 10-2009-0109693, filed on November 13, 2009, in the Korean Intellectual Property Office, the disclosure of which is incorporated herein in its entirety by reference.

The present invention relates to a method of preparing an extract fraction enriched with a particular component from Ginseng Radix, and more particularly, to a method of preparing an extract fraction enriched with ginsenoside Rg3 among saponins present in Ginseng Radix, ginsenoside Rg3 having vasodilatation activity and anti-cancer activity from Ginseng Radix.

Panax ginseng C.A. Mayer is a plant belonging to the Araliaceae family. Ginseng Radixes have been used in China as medicine from B.C. In Korea, Ginseng Radixes have been used in trading and as medicine from the period of the Three Kingdoms of Korea. Nowadays, they are widely used as herb medicine and health food in various fields.

Ginseng Radixes include as a species specific component about 3 to 6% of ginsenosides, which are saponin analogues, and the ginsenosides are a representative bio-active material of Ginseng Radix. According to the chemical structure, ginsenosides can be classified into panaxadiol(PD) saponin, panaxatriol(PT) saponin, and oleanane saponin. Currently, about 33 kinds of ginsenosides are known. The amount of saponins contained in ginseng radixes varies according to location, and some kinds of saponins are present in a very small amount. Thus, it is difficult to perform appropriate research regarding each kind of saponin.

PD saponins and PT saponins show different activities in the human body. PD saponins, for example, ginsenoside Rb1, inhibit a central nerve and are effective for mental stability, neurolepsis, pain release, antispasmodic decrease, blood-pressure decrease, and papaverine-like activity. PT-based saponins, for example, ginsenoside Rg1, are excitatory with respect to a central nerve and have anti-fatigue activity. In addition, ginsenoside Rg3 is present in a very small amount in white ginseng radixes. However, an amount of ginsenoside Rg3 contained in red ginseng radixes is 50 times greater than that in white ginseng radixes. Ginsenoside Rg3 has vasodilatation activity [J. Nat. Prod. 63, 1702 (2000)], platelet aggregation inhibition activity [Biol. Pharm. Bull. 21, 79(1998), Korean J. Ginseng Sci. 21, 132 (1997)], a brain cell protection activity [J. Neuroscience Res. 53, 426 (1998), Neuro Report 9, 226 (1998)], and anti-cancer activity [Jpn. J. cancer Res. 87, 357 (1996), J. cancer Res. Clin. Oncol. 120, 24 (1993), Anticancer Res. 17, 1067 (1997), Cancer Letters 150, 41 (2000), Dietary Anticarcinogenesis and Antimutagenesis 274 (2000)].

Generally, Ginseng Radix Rubra is produced as follows. First, fresh ginseng radixes are washed with water and placed in a container. Then, the fresh ginseng radixes are steamed with heated water vapors for a predetermined time period that may vary according to their size. Then, the ginseng radixes are dried with a hot air current and then sun-dried or dried using other drying methods until the water content therein is in the range of 12.5 to 13.5%. Due to the steaming and drying, a malonyl group contained in malonyl ginsenoside is removed, a glycosyl group is separated, and a hydroxyl group is isomerized, thereby producing various saponin components which are specific to Ginseng Radix Rubra. However, the production time is too long and the production costs are high. These factors prevent the widespread supply of Ginseng Radix Rubra.

Accordingly, if an extract enriched with ginsenoside Rg3 can be obtained from a ginseng radix, medicine or health food in which an medical activity of ginsenoside Rg3 is enhanced may be developed, and also, ginsenoside Rg3 may be economically and easily used without the complicated process for producing Ginseng radix Rubra.

KR Patent Publication No. 1999-0000458 discloses a method of producing ginsenoside Rg3 by reacting a diol-type saponin with lactase from Aspergillus sp.or Penicillium sp. in an aqueous solvent.

KR Patent Publication No. 2008-0063049 disclose a method of producing a Ginseng Radix extract or a Ginseng Radix Rubra extract enriched with ginsenoside Rg3, in which lactic acid bacteria are inoculated into Ginseng Radix or Ginseng Radixes Rubra and then incubated, and the incubation product is heat treated to convert an original type of ginsenoside of the Ginseng Radix or Ginseng Radix Rubra to ginsenoside Rg3.

KR Patent No. 0444394 discloses a method of producing an extract having a high amount of saponin by adsorption using an adsorption resin, but does not disclose a method of obtaining an extract fraction enriched with a specific saponin component.

Since the conventional methods for enriching ginsenoside Rg3 are performed using a microorganism or a microorganism-derived enzyme, the production process is complicated and expensive. Accordingly, there is a need to develop a simple chemical method of producing in high yield a ginseng radix extract fraction enriched with pure ginsenoside Rg3.

Accordingly, the inventors of the present invention found a condition when a great amount of ginsenoside that can be a reactant for producing ginsenoside Rg3 from ginseng radixes is obtainable, and developed a method of converting the ginsenoside into ginsenoside Rg3 to obtain pure ginsenoside Rg3 in high yield.

The present invention provides a method of preparing in high yield a ginseng radix extract fraction enriched with pure Rg3 by using a simple chemical process.

The present invention also provides a ginseng radix extract fraction enriched with Rg3 produced by using the method.

SUMMARY OF THE INVENTION

According to an aspect of the present invention, there is provided a method of preparing a ginseng radix extract fraction enriched with ginsenoside Rg3, the method including: enriching with ginsenoside Rg3 a ginsenoside Rb1-containing Ginseng Radix ethanol extract by reacting the ethanol extract with a mixture of water and acetic acid; adsorbing the ethanol extract enriched with ginsenoside Rg3 to an adsorption resin; allowing distilled water to go through the adsorption resin to elute components that are not adsorbed; adding 30 to 50 v/v% ethanol to the adsorption resin to remove non-saponin components and triol ginsenosides except for ginsenoside Rg3; and adding 50 to 100 v/v% ethanol to the adsorption resin to obtain an eluate.

According to another aspect of the present invention, there is provided a ginseng radix extract fraction enriched with ginsenoside Rg3 prepared by using the method, wherein the amount of the ginsenoside Rg3 is in a range of 200 to 500 mg/g.

DETAILED DESCRIPTION OF THE INVENTION

Hereinafter, the present invention will be described in detail.

The inventors of the present invention found that when an ethanol extract of a ginseng radix is reacted with a mixture including water and acetic acid, diol ginsenosides are converted to ginsenoside Rg3 in high yield. In addition, they also found that when the ginseng radix ethanol extract enriched with ginsenoside Rg3 is allowed to go through an adsorption resin and then 30-50 v/v% ethanol is added thereto for elution, non-saponin components and other saponin components except for ginsenoside Rg3 are removed and a ginseng radix extract enriched with pure ginsenoside Rg3 is obtained. Accordingly, an aspect of the present invention provides a method of preparing a ginseng radix extract fraction enriched with ginsenoside Rg3, the method including: enriching with ginsenoside Rg3 a ginsenoside Rb1-containing Ginseng Radix ethanol extract by reacting the ethanol extract with a mixture including water and acetic acid; adsorbing the ethanol extract enriched with ginsenoside Rg3 to an adsorption resin; allowing distilled water to go through the adsorption resin to elute components that are not adsorbed; adding 30 to 50 v/v% ethanol to the adsorption resin to remove non-saponin components and triol ginsenosides except for ginsenoside Rg3; and adding 50 to 100 v/v% ethanol to the adsorption resin to obtain an eluate.

The ethanol extract of a ginseng radix may be any one of various ginsenoside Rb1-containing extracts that are obtained by treating a ginseng radix with ethanol, for example, an extract including a high amount of ginsenoside Rb1. The ginseng radix ethanol extract may include, in addition to ginsenoside Rb1, a ginsenoside that is convertable to ginsenoside Rg3. Examples of the ginsenoside that is convertable to ginsenoside Rg3 include ginsenosides Rb2, Rc, and Rd, which are diol-type saponins, and a mixture thereof. An extract including a high amount of a diol-type ginsenoside, such as ginsenoside Rb1, may be obtained by adding 60-80% ethanol to a ginseng radix and extracting an extract at a temperature of 40 to 80℃ for 24 to 36 hours while agitating the mixture. This method is developed by the inventors as a method of obtaining an extract including a high amount of diol-type ginsenoside such as ginsenoside Rb1. If the extraction temperature is higher than 80℃, thermal stability of ginsenoside Rb1 is lowered and thus the amount of ginsenoside Rb1 obtained by extraction may be reduced. In addition, if the extraction temperature is lower than 40℃, the extraction efficiency may be decreased. Meanwhile, if the extraction time is outside the range described above and too long, economic efficiency or ginsenoside stability may be decreased, and if the extraction time is too short, the extraction efficiency may be reduced. For example, the ginseng radix ethanol extract may be an ethanol extract obtained by adding 65-75 v/v% ethanol to a ginseng radix and stirring them, at a temperature of 65-75℃ for 24-36 hours, or an ethanol extract obtained by adding 70 v/v% ethanol to a ginseng radix and stirring them at a temperature of 70℃ for 24 hours. The extraction process may be repeatedly performed three times or more and the obtained extracts are mixed up and the extract mixture is used as the ginsenoside Rb1-containing ginseng radix ethanol extract.

Then, the obtained ginsenoside Rb1-containing ginseng radix ethanol extract is reacted with a mixture including water and acetic acid so as to enrich the ginseng radix ethanol extract with ginsenoside Rg3. The ginsenoside Rb1, Rb2, Rc, or Rd contained in the ginseng radix ethanol extract is converted to ginsenoside Rg3 by reacting it with a mixture including water and acetic acid at a temperature of 70-90℃ for 24-36 hours. If the conversion reaction temperature is higher than 90℃, thermal stability of ginsenoside Rb1 is lowered, and correspondingly, less ginsenoside Rg3 is formed. In addition, if the conversion reaction temperature is lower than 70℃, the conversion efficiency may be reduced. Meanwhile, if the conversion time is outside the range described above and too long, the economic efficiency or ginsenoside stability may be decreased, and if the conversion time is too short, the conversion efficiency may be reduced. For example, the reaction may be performed at a temperature of 80-90℃ for 24-36 hours. A ratio of water and acetic acid in the mixture may not be limited as long as the mixture including water and acetic acid can convert the reactant diol-type ginsenoside into ginsenoside Rg3. However, in order to produce ginsenoside Rg3 in high yield, based on 100 parts by weight of ethanol, the amount of water included in the mixture may be in a range of 30 to 70 parts by weight and the amount of the acetic acid included in the mixture may be in a range of 30 to 70 parts by weight. Compared to a mixture including ethanol, water, and acetic acid, a mixture of water and ethanol, or a mixture of acetic acid and ethanol, when the mixture of water and acetic acid is used, the diol-type ginsenoside such as ginsenoside Rb1 may be converted to ginsenoside Rg3 in high yield. For example, a mixture of water and acetic acid in a mixture ratio of 3:1 is added to a ginsenoside Rb1-containing ginseng radix ethanol extract and the reactants are reacted at a temperature of 90℃ for 24 hours so as to convert the ginsenoside Rb1 to ginsenoside Rg3, in which a volume ratio of the mixture to the ginsenoside Rb1-containing ginseng radix ethanol extract is 10:1.

Then, the ginseng radix ethanol extract enriched with ginsenoside Rg3 is introduced to an adsorption resin so as to isolate a ginsenoside Rg3 fraction. Before the adsorption process, the ginseng radix ethanol extract enriched with ginsenoside Rg3 may be concentrated to a 70 to 90 brix and then diluted with water, for example, distilled water. The dilution may be performed to such an extent that a concentrate of the ethanol extract enriched with ginsenoside Rg3 is reacted with and saponins are adsorbed to the adsorption resin. The amount of the water used for the dilution may not be limited, and may be about four to six times or about five times greater than a weight of the concentrated extract. The adsorption resin may be Diaion HP-20 (Mitsubishi, Japan), and may be used in the form of a column filled for adsorption and fraction.

Then, a component that is not adsorbed to the adsorption resin is eluted out and removed by allowing water, for example, distilled water, to go through the adsorption resin. Thus, the component that is not adsorbed to the adsorption resin is eluted out. The amount of the distilled water used to remove the component that is not adsorbed to the adsorption resin may not be limited as long as a non-saponin component is removable by elution, and for example, may be about 5 times greater than the volume of the adsorption resin.

Then, in order to remove the non-saponin component and triol-based ginsenosides, except for ginsenoside Rg3, adsorbed to the adsorption resin, 30 to 50 v/v% ethanol is added to the adsorption resin to elute the non-saponin component and triol-based ginsenosides except for ginsenoside Rg3. In an embodiment, before 30 to 50 v/v% ethanol is added to the adsorption resin to elute the non-saponin component and triol-based ginsenosides except for ginsenoside Rg3, 0 to 30 v/v% ethanol may be added to the adsorption resin to perform an elution process. Through the processes described above, most other components excluding ginsenoside Rg3 are removed from the adsorption resin. Accordingly, in the subsequent process, ginseng radix extract fraction enriched with pure ginsenoside Rg3 may be obtained.

When the non-saponin component and the triol-based ginsenosides except for ginsenoside Rg3 are removed from the adsorption resin, an elution process is performed by adding 50 to 100 v/v% ethanol to the adsorption resin, thereby obtaining a ginseng radix extract fraction enriched with ginsenoside Rg3. In the ginseng radix extract fraction enriched with ginsenoside Rg3 obtained as described above, the amount of ginsenoside Rg3 may be in the range of 100 to 200 mg/g.

The ginseng radix used to prepare the ginseng radix extract may be a ginseng radix that includes a diol-type ginsenoside which is convertible to ginsenoside Rg3, for example, ginsenoside Rb1, and examples of such a ginseng radix include a white ginseng radix, a fresh ginseng radix, a ginseng radix rubra, a Taekuk ginseng radix, black ginseng radix, a dextrinization ginseng radix (puffing ginseng radix), and an enzyme-treated ginseng radix or concentrate thereof.

According to another aspect of the present invention, a ginseng radix extract fraction enriched with 200 to 450 mg/g of ginsenoside Rg3 prepared according to the method described above is provided.

As described above, according to the above embodiments of the present invention, a ginseng radix extract fraction enriched with pure ginsenoside Rg3, which is one of saponin components included in Ginseng Radix in a small amount and is present in Ginseng Radix Rubra more than in Ginseng Radix, may be prepared using a simpler chemical method compared to a conventional method using a microorganism.

FIG. 1 shows a high performance liquid chromatography (HPLC) chromatogram of an ethanol extract of a red ginseng radix obtained by adding 70% ethanol to 1 kg of the red ginseng radix in such an amount that the weight of ethanol was 15 times greater than that of ginseng radix rubra, performing an extraction process three times, and then filtering the extraction product.

FIG. 2 shows a HPLC chromatogram of a ginseng radix ethanol extract obtained by extracting ginseng radix palva with 70 v/v% ethanol at a temperature of 70℃ for 24 hours.

FIG. 3 shows a HPLC chromatogram of fractions obtained in such a manner that a mixed solvent including water and an acetic acid in a ratio of 3:1 was added to ginseng radix ethanol extract obtained by extracting ginseng radix palva with 70 v/v% ethanol at a temperature of 70℃ for 24 hours and the mixture was reacted at a temperature of 90℃ for 24 hours and then the reaction product was concentrated and diluted and adsorbed to the adsorption resin HP-20, and then extraction was performed thereon with 0-30 v/v% ethanol.

FIG. 4 shows a HPLC chromatogram of fractions obtained in such a manner that a mixed solvent including water and an acetic acid in a ratio of 3:1 was added to ginseng radix ethanol extract obtained by extracting ginseng radix palva with 70 v/v% ethanol at a temperature of 70℃ for 24 hours and the mixture was reacted at a temperature of 90℃ for 24 hours and then the reaction product was concentrated and diluted and adsorbed to the adsorption resin HP-20, and then extraction was performed thereon with 0-30 v/v% ethanol and then 30-50 v/v% ethanol.

FIG. 5 shows a HPLC chromatogram of fractions obtained in such a manner that a mixed solvent including water and an acetic acid in a ratio of 3:1 was added to ginseng radix ethanol extract obtained by extracting ginseng radix palva with 70 v/v% ethanol at a temperature of 70℃ for 24 hours and the mixture was reacted at a temperature of 90℃ for 24 hours and then the reaction product was concentrated and diluted and adsorbed to the adsorption resin HP-20, and then extraction was performed thereon with 0-30 v/v% ethanol, 30-50 v/v% ethanol, and then 50-100 v/v%.

The present invention will be described in further detail with reference to the following examples. These examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

Comparative Example 1: Preparation of Ginseng Radix Rubra Extract

70% ethanol was added to 1 kg of ginseng radix rubrain which the weight of ethanol was 15 times greater than that of ginseng radix rubra. Then, an extraction process was repeatedly performed three times and the obtained three extracts were mixed up and filtered to prepare an ethanol extract of the red ginseng radix. The ethanol extract of the red ginseng radix was analyzed by high performance liquid chromatography (HPLC). The obtained HPLC chromatogram is illustrated in FIG. 1.

Example 1: Preparation of Ginseng Radix Extract using 70 v/v% ethanol

70% ethanol was added to 1 kg of ginseng radix palva, in which the weight of 70% ethanol was 15 times greater than that of the ginseng radixes, and an extraction process was repeatedly performed thereon three times while an extraction condition was changed as shown in Table 1, and the obtained three extracts were mixed up and filtered, thereby preparing ginseng radix ethanol extracts. With regard to each of the ginseng radix ethanol extracts, the amount of the ginsenoside Rb1 was measured by HPLC. The results are shown in Table 1.

Table 1

Extraction Conditions (temperature-hour)	Rb1 (mg/g)	Extraction Conditions (temperature-hour)	Rb1 (mg/g)
40℃ - 3hr	9.928	70℃ - 3hr	11.098
40℃ - 6hr	10.182	70℃ - 6hr	13.001
40℃ - 10hr	10.799	70℃ - 10hr	16.127
40℃ - 24hr	11.572	70℃ - 24hr	20.186
50℃ - 3hr	10.698	70℃ - 30hr	20.904
50℃ - 6hr	10.881	70℃ - 36hr	21.405
50℃ - 10hr	11.404	80℃ - 3hr	14.957
50℃ - 24hr	13.607	80℃ - 6hr	15.598
60℃ - 3hr	10.852	80℃ - 10hr	17.254
60℃ - 6hr	11.718	80℃ - 24hr	17.841
60℃ - 10hr	12.744	80℃ - 30hr	16.776
60℃ - 24hr	16.588	80℃ - 36hr	15.836

From the experimental results, it can be seen that when the extraction process was performed using 70 v/v% ethanol, the amount of ginsenoside Rb1 depended very much on the extraction temperature. In particular, the amount of ginsenoside Rb1 was very high at a temperature of about 70℃. In addition, when the extraction time was 24 hours or more, the amount of ginsenoside Rb1 was high. However, at a temperature of 80℃, the amount of ginsenoside Rg3 decreased as the extraction time increased. This result may be due to temperature instability.

It can be seen that it is desired to prepare the ginseng radix ethanol extract at a temperature of 70℃ for 24 hours. The ethanol extract obtained at a temperature of 70℃ for 24 hours was analyzed by HPLC and the obtained HPLC results are shown in FIG. 2.

Example 2: Ginsenoside Rg3 Enrichment Process

A concentrate (75-80 brix) of the ginseng radix ethanol extract prepared at a temperature of 70℃ for 24 hours in Example 1 was mixed with a mixed solvent shown in Table 2 below (10 times greater than an amount of the concentrate), and the resultant solution was heated at a temperature of 90℃ for 24 hours. The amount of ginsenoside Rg3 contained in the reaction product was measured by HPLC. The results are shown in Table 2.

In addition, the same concentrate of the ginseng radix ethanol extract was reacted in the same manner as described above and under the conditions shown in Table 3 below, and then the ginsenoside Rg3 was measured by HPLC. The results are shown in Table 3.

Table 2

Extraction results according to used solvent (heating at 90 ℃ for 24 hours)

Ginseng Radix Concentrate Extraction Conditions	Rg3 content(mg/g)
100 % water	11.216
70 % ethanol	12.615
Water:acetic acid=3:1	23.520
Water:ethanol:acetic acid=2:1:1	23.272

Table 3

Extraction results with time Unit: mg/g

Ginseng Radix Concentrate Extraction Conditions	24 hr. extraction		48 hr.extraction
Ginseng Radix Concentrate Extraction Conditions	55 ℃	90 ℃	55 ℃	90 ℃
100 % water	1.528	11.216	2.375	14.816
70 % ethanol	1.753	12.615	4.534	13.467
Water:acetic acid=3:1	6.331	23.520	14.448	22.536
Water:ethanol:acetic acid=2:1:1	4.768	23.272	9.297	21.200

From the experimental results, it can be seen that when the mixture of water and acetic acid was added to the ginseng radix ethanol extract, ginsenoside Rg3 was obtained by conversion in a substantially high yield compared to when ethanol, water, and acetic acid were used separately or a combination of ethanol and acetic acid was used. In addition, it can also be seen that when the mixture of water and acetic acid in a mixture ratio of 3:1 was used and the heating was performed at a temperature of 90℃ for 24 hours, ginsenoside Rg3 was obtained by conversion in a substantially high yield, compared to other temperature and time conditions.

Example 3: Adsorption Process using Adsorption Resin Diaion HP-20

The reaction product including ginsenoside Rg3 prepared by using the mixture of water and acetic acid in a mixture ratio of 3:1 and heating at a temperature of 90℃ for 24 hours according to Example 2 was concentrated to produce 65 brix of a concentrate. Then, the concentrate was sufficiently diluted with water in a weight of 5 times greater than that of the concentrate, and then, the diluted concentrate was allowed to go through an adsorption resin Diaion HP-20, so that saponin components were adsorbed to the adsorption resin. Then, distilled water in an amount of 5 times greater than the volume of the adsorption resin was continuously allowed to go through the adsorption resin to remove components that were not adsorbed to the adsorption resin.

In order to remove a non-saponin component and triol saponin components except for ginsenoside Rg3 from the adsorption resin, 0 to 30 v/v% ethanol was allowed to go through the adsorption resin and an ethanol elution fraction was obtained, and then 30 to 50 v/v% ethanol was allowed to go through the adsorption resin and an ethanol elution fraction was obtained.

Then, 50 to 100 v/v% ethanol was allowed to go through the adsorption resin and an ethanol elution fraction was obtained. In this regard, the amount of ethanol used for the elution processes described above was 5 times greater than the volume of the adsorption resin at each step. Each ethanol elution fraction was analyzed by HPLC and the results are respectively shown in FIG. 3 (0 to 30 v/v% ethanol elution fraction), FIG. 4 (30 to 50 v/v% ethanol elution fraction), and FIG. 5 (50 to 100 v/v% ethanol elution fraction).

Referring to FIGS. 3 to 5, the 0 to 30 v/v% ethanol elution fraction and the 30 to 50 v/v% ethanol elution fraction did not include ginsenoside Rg3, and the 50 to 100 v/v% ethanol elution fraction that was obtained after the 0 to 30 v/v% ethanol elution and the 30 to 50 v/v% ethanol elution were performed, included ginsenoside Rg3, which was identified by presence of a corresponding peak in FIG. 3.

Experimental example 1: Evaluation of Amount of Ginsenoside Rg3 included

The amounts of ginsenoside Rg3 in the extracts prepared according to Examples 1 through 3 were evaluated by HPLC. In regard to Example 1, the amount of ginsenoside Rg3 in the ethanol extract prepared at a temperature of 70℃ for 24 hours was evaluated; in regard to Example 2, the amount of ginsenoside Rg3 in the product obtained by adding a mixture of water and acetic acid in a mixture ratio of 3:1 to the ethanol extract and heating the solution at a temperature of 90℃ for 24 hours to convert to ginsenoside Rg3 was evaluated; and in regard to Example 3, the amount of ginsenoside Rg3 in the 50 to 100 v/v% ethanol elution fraction was evaluated.

The evaluation results are shown in Table4.

Table 4

	amount of Rg3 (%)	amount of Rg3 (mg/g)
Ethanol extract of ginseng radix (Example 1)	-	-
Reaction product obtained by converting to ginsenoside Rg3 in ginseng radix ethanol extract (Example 2)	2.2-2.4 %	22-24 mg/g
50 to 100 v/v% ethanol elution fraction from Adsorption resin(Example 3)	26-44 %	260-440 mg/g

Referring to the results shown in Table 4, although ginsenoside Rg3 was not originally present in the ginseng radix ethanol extract, the amount of ginsenoside Rg3 in the 50 to 100 v/v% ethanol fraction eluted from the adsorption resin after the conversion reaction to ginsenoside Rg3, was high. Thus, it can be seen that the final product was a ginseng radix extract fraction enriched with ginsenoside Rg3.

In addition, compared to the Ginseng Radix Rubra ethanol extract prepared in Comparative Example 1, a ginseng radix extract fraction prepared using the method according to the present invention includes a high amount of ginsenoside Rg3.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the present invention as defined by the following claims.

Claims

A method of preparing a ginseng radix extract fraction enriched with ginsenoside Rg3, the method comprising:

enriching with ginsenoside Rg3 a ginsenoside Rb1-containing Ginseng Radix ethanol extract by reacting the ethanol extract with a mixture of water and acetic acid;

adsorbing the ethanol extract enriched with ginsenoside Rg3 to an adsorption resin;

allowing distilled water to go through the adsorption resin to elute components that are not adsorbed;

adding 30 to 50 v/v% ethanol to the adsorption resin to remove non-saponin components and triol ginsenosides except for ginsenoside Rg3; and

adding 50 to 100 v/v% ethanol to the adsorption resin to obtain an eluate.
The method of claim 1, wherein the ginsenoside Rb1-containing Ginseng Radix ethanol extract is prepared by adding 60-80 v/v% ethanol to the ginseng radix and stirring them at a temperature of 65 to 75℃ for 24 to 36 hours.
The method of claim 1, wherein the mixture including water and acetic acid contains water in a range of 60 to 80 parts by weight and acetic acid in a range of 20 to 40 parts by weight.
The method of claim 1, wherein, in enriching with ginsenoside Rg3 the ginsenoside Rb1-containing Ginseng Radix ethanol extract, the ethanol extract is reacted with the mixture of water and acetic acid at a temperature of 80 to 90℃ for 24 to 36 hours.
The method of claim 1, wherein the Ginseng Radix ethanol extract enriched with ginsenoside Rg3 is concentrated to 70-90 brix and then diluted with distilled water and added to the adsorption resin.
The method of claim 1, wherein the adsorption resin is a Diaion HP-20.
The method of claim 1, wherein the ginseng radix is a white ginseng radix, a fresh ginseng radix, a ginseng radix rubra, a Taekuk ginseng radix, black ginseng radix, a dextrinization ginseng radix (puffing ginseng radix), or an enzyme-treated ginseng radix or concentrate thereof.
A ginseng radix extract fraction enriched with ginsenoside Rg3 prepared by using any one of the methods of claims 1-7, wherein the amount of the ginsenoside Rg3 is in a range of 200 to 500 mg/g.