KR101927417B1 - Method for converting and producing functional saponin from ginseng extract using Subcritical Water and composition using the same - Google Patents

Abstract

The present invention relates to a method for converting ginseng saponin to a specific saponin using ginseng extract obtained from an extracting solvent (such as water or ethanol) from ginseng (ginseng, white ginseng, red ginseng, etc.) (Rg2, Rg3, Rh1, Rh2, Rg3, Rg3, and Rg3), which have high absorption capacity in human body and excellent pharmacological activity, are treated with ginseng extract for an appropriate period of time with appropriate sublimation factor having appropriate conditions Rg5, Rk1, Rk3, PPT, PPD, etc.). The specific saponin conversion method of the present invention is an epoch-making method which can maximize the content of the specific saponin as compared with the existing technology, and has the advantages of high production efficiency, reduced manufacturing time and process steps, and excellent economical efficiency.

Description

TECHNICAL FIELD The present invention relates to a method for converting and producing functional saponins from ginseng extract using sub-critical water and a composition using the same.

The present invention relates to a method for efficiently producing ginseng saponin by using sub-critical water and converting it into a specific saponin form possessed by red ginseng. More specifically, (Rg3, Rg5, Rk1, Rk3, Rh2 and the like) known to have excellent physiological activity by using ginseng saponin in the form of a glycoside present in the ginseng extract itself extracted by a conventional method (water or alcohol) , And a method of dramatically increasing the content and concentration of the specific saponin.

Ginseng has been used as an indicator of mystery in the Orient for 2,000 years due to its excellent pharmacological effect. The main functional ingredient of ginseng is ginseng saponin, which is called 'ginsenoside', only the ginseng saponin among the various saponins of the plant. Ginseng saponin is a glycoside in which saccharides such as glucose, rhamnose, xylose and arabinose are ester-bonded to alcoholic OH groups of R1, R2 and R3 of aglycon having a dammarane skeleton (glycosides: glycosides).

Ginsenoside saponins are predominantly composed of ginsenosides Rb1, Rb2, Rc and Rd belonging to protopanaxadiol saponin and ginsenosides Rg1, Re and Rf belonging to protopanaxatriol saponin .

When ginseng saponin is absorbed in the body, it is decomposed by the microorganisms in the human intestine and absorbed into the human body. However, intestinal microorganisms that degrade ginseng saponin vary depending on the presence and the degree of retention of the microorganism.

It is known that ginseng saponin is the main component showing the pharmacological effect of ginseng, but the components which act substantially on the anti-cancer action, suppression of cancer metastasis, antiallergic, blood circulation improvement and dementia prevention are Rg3, Rg5, Rk1, Rh2, Rk3, and the like.

The specific saponin was originally produced by the separation of the sugar from the ginsenosides such as ginsenosides Rb1, Rb2, Rc, Rd, Rg1, Re, and Rf in ginseng and dehydration reaction. Absorption is excellent in the intestines without.

On the other hand, red ginseng is a heat-processed processed food that can be stored for a long period of time by steam-drying the ginseng with steam. During the steaming process of ginseng, the starch particles in the ginseng are enriched and the digestibility is higher than that of white ginseng. It is advantageous for storage, and chemical conversion by hydrolysis occurs during the production process, so that a new physiologically active ingredient (specific saponin) which does not exist in ginseng and white ginseng is produced and content is increased. Studies have shown that ginsenosides Rg3, Rg5, Rk1, and Rh2 (specific saponins) found in red ginseng have excellent pharmacological activities in anticancer effects, anti-cancer effects, antiallergic effects, immune enhancement, Has been widely reported and the excellent pharmacological activity of a small amount of specific saponin has been verified and studies for the production of these specific saponins have been actively carried out. The major saponins are mainly processed by heat treatment, acid / alkali hydrolysis, enzymatic hydrolysis, microbial fermentation Hydrolysis, organic synthesis, etc. As a production method using acid hydrolysis, Korean Patent No. 10-0635025 discloses a method in which brewing vinegar having a pH of 2.27 to 2.9 is added and then heated and extracted for 0.5 to 24 hours Rg3, Rg5 and Rh1, and Korean Patent No. 10-0453542 discloses a method for producing a mixture of Rg3 and Rg5 Korean Patent Registration No. 10-0471340 discloses a method of adding ginseng to a specific herbal medicine containing a large amount of organic acids such as citric acid instead of an organic acid such as citric acid and heating the mixture at a boiling temperature to obtain Rg3, Rg5, Rk1 And a processing method for increasing the content is proposed. However, the acid hydrolysis method has a disadvantage in that the selectivity in the hydrolysis process is deteriorated, and the effect of the edible product as an edible product which does not remove the added acid is deteriorated.

The specific saponin components (Rg3, Rg5, and Rg5) were heat-treated at 110 to 180 degrees, which is much higher than the red ginseng production temperature, for 0.5 to 20 hours in Korean Patents 10-0192678, 10-0228510 and 10-0555652, Etc.), and currently, it is sold as ginseng processed product called 'ginseng (ginseng)'.

However, since this method is heated by steam (high-temperature high-pressure steam) in a pressure vessel such as a high-pressure sterilizer, the penetration of water molecules and the hydrolysis reaction rate into the ginseng sample are slower than the subcritical water used in the present invention, Conversion to specific saponin is not only required for a long period of time but also has a disadvantage in that the specific saponin conversion efficiency is low and the ginseng sample is directly contacted to the reactor to cause carbonization.

Further, in order to increase the content and increase the conversion to specific saponin, Korean Patent Nos. 10-0949247, 10-10-0921257, 10-0877489, 10-0443411, 10 -0412218, and the like, and it is also known that Korean Patent No. 10-0813997, 10-0811295, 10-0856790, 10-0935362, etc., which make fermented ginseng using microorganisms such as intestinal bacteria, lactic acid bacteria, fungi, , 10-0050413, 10-0762154, 10-0899837, etc. However, these methods have a problem in that the yield of specific saponin conversion (production) is low, and they have disadvantages of producing only specific saponin.

SUMMARY OF THE INVENTION The present invention has been made to solve the above problems, and it is an object of the present invention to provide a method for enhancing the content of specific saponins having excellent pharmacological activity and excellent in body absorption rate.

Another object of the present invention is to provide a specific saponin produced by the above method

In order to accomplish the above object, the present invention provides a method for producing a specific saponin from ginseng extract comprising converting a ginseng extract into a specific saponin through a sub-critical water reactor.

In one embodiment of the present invention, the ginseng extract is preferably an extract of ginseng, white ginseng or red ginseng, but is not limited thereto.

In another embodiment of the present invention, the temperature of the sub-threshold water reactor is preferably 120 to 200 ° C, but is not limited thereto.

The sub-threshold water reactor is preferably operated under the condition of 0.20 MPa to 1.56 MPa considering the economical efficiency such as the device investment cost according to the high pressure, but it is preferable that the maximum pressure capable of maintaining the sub- It may be performed up to 22.1 MPa.

The amount of the sub-critical water required for the reaction is preferably 20 times or more the amount of the ginseng extract solid content, but is not limited thereto.

In another embodiment of the present invention, the specific saponin is preferably saponin selected from the group consisting of Rg3, Rk1, Rg5, Rh2, Rk3 protopanaxadiol, Rg2, Rh1, Rh4 and protopanaxatriol But not limited to this.

The present invention also provides a composition comprising the specific saponins produced by the method of the present invention.

Hereinafter, the present invention will be described.

The present invention is a method which utilizes sub-critical water of an optimal condition (compressed hot water subjected to pressure) in a different method from the prior art.

More specifically, the water in the subcritical state has a method of converting ginseng saponin into a specific saponin by utilizing the property of causing hydrolysis reaction due to a remarkably high dissociation constant of hydrogen ion [H ⁺ ] and hydroxide ion [OH ^- ] (Rg3, Rk1, Rg5, Rh2, Rk3, etc.) having excellent pharmacological activity and high in-body absorption rate.

The sub-threshold number will be briefly described below.

The critical point of the water is 374 ° C and the pressure is 22 MPa. The water above the critical point is called supercritical water. Subcritical water is pressurized hot water or compressed hot water. Referred to as superheated water, refers to hot water that maintains a liquid state in a state where a pressure is applied between a boiling temperature (100 ° C) and a critical temperature (374 ° C) of water (see FIG. 1).

As the temperature of water increases, the physicochemical properties of water, especially the relative dielectric constant and ion product,

Relative dielectric constant of water [ e . Or polarity of water] is significantly reduced at 25 ° C as the temperature increases from 80 to 250 ° C to 27 ° C. This means that the water in the subcritical state at 250 ° C has polar and nonpolar properties such as methanol ( e = 33) and ethanol ( e = 24) at 25 ° C, and thus has the ability to extract (dissolve). The ionic mass (Kw) of water is the product of hydrogen ion (H +) and hydroxide ion (OH-) concentration, which can be expressed as ion product of water or the dissociation constant for water.

At 1 atm at room temperature, the ionic value of water is 10 ^-14 , which increases abruptly with increasing temperature. It increases to 10 ^-11 at 25 MPa, at 250 ℃, and decreases to 10 ^-19 at 390 ℃. , And decreases to 10 ^-22 at 500 ° C.

That is, the dissociation constants of hydrogen ions and hydroxide ions in the subcritical water are increased to three times the size of the water at room temperature and pressure (1000 times), which is the acidity or basicity Means that it can act as a catalyst for an acid or a base without adding a substance, and has a high hydrolytic capacity (see FIG. 2).

The specific saponin was originally produced by the separation of sugar from the ginsenosides such as ginsenosides Rb1, Rb2, Rc, Rd, Rg1, Re, and Rf in ginseng and dehydration reaction.

More specifically, the specific saponins belonging to the protopanaxadiol system include Rg3, Rk1, Rg5, Rh2, and PPD. Rg3 is linked to the C-20 position of aglycone in Rb1, Rb2, Rc, Rg1 and Rg5 were produced by the dehydration (-H2O) at C-20/21 and C-20/22 positions of aglycone at Rg3, and Rh2 was generated by the hydrolysis of C- One of the sugars linked to the 3-position is formed by hydrolysis. When all of the sugars connected to the C-3 position of aglycone in Rh2 are hydrolyzed, PPD becomes protopanaxadiol.

The specific saponins belonging to the protopanaxatriol system include Rg2, Rh1, Rk3, Rh4 and PPT. Rg2 is generated by hydrolysis of sugars connected to the C-20 position of aglycone in Re, and Rh1 20 is produced by hydrolysis of the sugar linked to the C-20 position of aglycone at Rg1, or hydrolysis of one sugar linked to the C-6 position of aglycone at Rf, while Rk3 and Rh4 are produced by hydrolysis of C- 21 and C-20/22, and PPT (protopanaxatriol) is formed by hydrolysis of all of the sugars connected to the C-6 position in Rh1. (See the structure of the ginsenoside of Fig. 3 and the conversion of ginsenoside of Fig. 4)

In order to convert ginseng saponin into a specific saponin as described above, a hydrolysis reaction process such as heat treatment, acid treatment, enzyme treatment, and fermentation with microorganisms is indispensable.

In view of the fact that the water in the subcritical state can accelerate the hydrolysis reaction by a remarkable increase in the ionic property, the present invention can be applied to a reaction using a sub-critical water having an optimal hydrolysis capacity for rapidly converting ginsenoside to a specific saponin (Temperature, pressure, concentration of extracted solid content, reaction time, and the like), thereby completing the present invention.

The present inventors have solved the problems of the present invention through the following various steps

Step 1: Crushing the ginseng or white ginseng to prepare ginseng extract by water or alcohol in a conventional manner;

Step 2: The step of making ginseng concentrate (extract) by vacuum distillation or the like of the ginseng extract of step 1;

Step 3: diluting the ginseng concentrate (extract) of step 2 with an appropriate amount of purified water (water);

Step 4: The diluted ginseng concentrate of step 3 is reacted with the appropriate temperature and pressure through a sub-critical water reactor for an appropriate time to convert it to ginsenoside;

Here, the sub-critical water reactor is a batch reactor or a continuous reactor, and it controls the temperature, the pressure and the reaction time in order to make the state of the water contained in the three-step diluted ginseng concentrate into the subcritical state It is a device that can do it.

Step 5: recovering the reacted extract of step 4;

- The extract is cooled and recovered in the form of an extract.

- The extract is recovered as a concentrate by a method such as distillation under reduced pressure as in Step 2

- Collecting the concentrate in powder form by spray drying (SD) or lyophilization (FD)

- Since the extract in the four-step reactor is in a high-temperature and high-pressure state, it can be recovered directly from the extract to the concentrate immediately after completion of the reaction process by flash vaporization through a spray nozzle connected to the reactor.

In the case of extracting with water in step 1 of the above step, it is possible to proceed directly to step 4 when step 1 is completed without going through steps 2 and 3. However, before entering step 4, the content of water is adjusted so that the ratio of the extracted solid content to water in the first step extract is suitable for the formation of the specific saponin conversion (when the water content is large)

If the water content is low, refined water should be replenished.

According to the present invention, first, the ginseng extract produced by the present invention has a high specific saponin content and a high concentration, so that the absorption rate in the human body can be increased regardless of the presence or absence of intestinal microorganisms and excellent pharmacological activity can be expected.

Second, since the content and concentration of the specific saponin can be maximized according to the present invention, it is possible to easily produce a product in which the content of ginsenoside is standardized (for example, (Rg3 + Rg5 + Rk1 + Rk3) 5% or more of solid content)

Third, the reaction process for producing specific saponin conversion according to the present invention can be continuously performed within 30 minutes,

Fourth, since the ginseng extract immediately after the reaction for producing specific saponin conversion is in a high-temperature and high-pressure state, it is possible to omit the concentration process that requires a large energy cost by flash evaporation through the injection nozzle in the reaction apparatus, Process can be constructed,

Fifth, the specific saponin in the present invention can be obtained without the red ginseng processing (steam cooking, drying, etc.) of ginseng (ginseng), shortening the manufacturing process and manufacturing time, and greatly reducing energy cost.

Figure 1 shows the phase diagram of water,
Figure 2 shows the ionic, relative dielectric constant, and density variations of water with temperature,
3 shows the structure of ginsenoside,
4 is a table summarizing the conversion of ginsenosides.

The present invention will now be described in detail with reference to the following non-limiting examples. However, the following examples are intended to illustrate the present invention, and the scope of the present invention is not to be construed as being limited by the following examples.

Example  1: Production of ginseng extract concentrate

Step 1: Preparation of ginseng extract and second step: Concentration of ginseng extract

The first step for carrying out the present invention is to obtain ginseng extract using a conventional method (hot water extraction or alcohol extraction (70% EtOH)) using fresh ginseng or white ginseng. Ginseng extract (extract) is made by vacuum distillation or other methods.

1,000 g of 4-year old fresh ginseng and 6,000 ml of 70% EtOH were placed in an extractor, and 5,654 ml of ginseng extract was obtained by vacuum extraction (extraction temperature 55 ° C, extraction time 8 hours). The extract was distilled under reduced pressure To obtain 103 g of a ginseng extract concentrate having a solid content of 85%.

Raw materials Extraction solvent Extract concentrate Extraction yield
(Solids / raw materials) Quantity Solids 1kg of fresh ginseng Alcohol (70%)
6,000ml 5,654 ml 103g 85% 8.76%

Gin Senocide Content (mg / g) Remarks division Name Dioscorpion system Ginseng saponin Rb1 13.03 Rb2 3.67 Rc 4.80 Rd 4.83 sub Total 26.33 Specific saponin Rg3 0.00 Rk1 0.35 Rg5 0.42 Rh2 1.24 PPD 0.14 sub Total 2.15 system 28.48 Tricolor Ginseng saponin Re 9.82 Rf 2.27 Rg1 8.11 sub Total 20.2 Specific saponin Rg2 0.73 Rh1 0.52 Rk3 0.29 Rh4 0.07 PPT 0.18 sub Total 1.79 system 22.0 sum ( Total saponin ) (A) 50.48 Specific saponin system (B) 3.94 Specific saponin ratio (B / A) 7.8%

In Table 2, * ginsenoside content (mg / g) refers to the content of each ginsenoside contained in 1 g of extracted solid,

 * PPD = protopanaxdiol, PPT = protopanaxtriol

* The analysis of ginsenoside content was carried out three times under the same conditions.

Regarding the ginsenoside analysis conducted in the present invention,

- Instrument: Agilent 1100 series

- Column: GraceSmart P18 5U 250mm × 4.6mm

- Mobile Phase: Water - Acetonitrile = 80 - 20 (gradient)

- Detector & Wavelength: UV 203nm

- Flow rate: 1.2 ml / min

- Injection Volume: Standard (20 μl), Sample (20 μl)

- Colume oven temp. : 40 ° C

Equipment and conditions were used,

The specific analysis method

The prepared sample was ultrasonically extracted with MeOH to dissolve it, filtered using a 0.45 μm syringe filter, and used as an analytical sample. The column used for the analysis was GraceSmart P18 5 U 250 mm × 4.6 mm. The flow rate was 1.2 ml / min. The gradient conditions were as follows: Water: ACN = 80:20 for 10 minutes, ACN 30% for 52 minutes, ACN 40% for 77 minutes , The ACN was flowed to 47% for 80 minutes, the ACN was flowed to 70% for 100 minutes, the ACN was flown to the same amount for 105 minutes, the ACN was flown to 20% for 106 minutes, Rg2, Rg3, Rg5, Rh2, Compound K, and PPD were used as the ginsenoside Rb1, Rb2, Rc, Rd, Rg3, , Rk1, Re, Rf, Rg1, Rg2, Rh1, F1, and PPT are derived from ChromaDex And Rh4 + Rk3 was purchased from Ambo Laboratories.

The purity of this standard is 85.8, 97.3, 84, 98.2, 95.7, 95, 97.9, 94.5, 94.7, 100, 96.7, 95.3, 95.9, 92.3, 91.4, 100, 97.6, 31.8 + However, since the purity of ginsenoside Rk1 and F1 standard is unknown to the manufacturer, it is regarded as 100% and reflected in the result.

Step 3/4: Subcritical  -Specific saponin conversion using water

As a key process of the present invention, the inventors of the present invention have conducted research and development and have found that the mixing ratio of ginseng extract solid content and water (subcritical water), reaction temperature, pressure, reaction time and the like affect the conversion of specific saponins We found that it was a major variable.

As for the temperature condition of subcritical water with respect to the specific saponin conversion reaction, ionic (dissociation of hydrogen ion [H +] and hydroxyl ion [OH-]) becomes remarkable at 200 ~ 250 ℃, When the reaction time is prolonged, the ginsenosides are decomposed and there is a problem in controlling the conversion of specific saponins.

(Less dissociation of hydrogen ions [H ⁺ ] and hydroxide ions [OH ^- ]) than at high temperatures at temperatures lower than 100 to 120 ° C, the conversion rate to specific saponins is slow, have.

Therefore, it is more advantageous to carry out a specific saponin conversion formation reaction at 120 to 200 ° C.

On the other hand, in the subcritical water at 120 to 200 ° C, the higher the pressure, the higher the ionic mass. Therefore, instead of lowering the temperature, the pressure may be increased (eg, 150 ° C, 20Mpa)

In order to apply high pressure, relatively expensive production equipment investment is required, and energy cost, process time extension due to pressurization and depressurization are inefficient. Therefore, it is controlled only by the pressure in the condition where sub-critical water (liquid) is maintained (E.g., 150 DEG C: 0.48 Mpa, 160 DEG C: 0.62 Mpa, 170 DEG C: 0.79 Mpa, 180 DEG C: 1.0 Mpa, 190 DEG C: 1.26 Mpa, 200 DEG C: 1.56 Mpa)

The conditions for the mixing ratio of ginseng extract and water are as follows.

The reaction rate (specific saponin conversion) tends to accelerate as the solids concentration is higher under the same reaction conditions (temperature, pressure), but the solids concentration The disadvantage is that it is difficult to control the reaction because the specific saponin conversion rate and conversion efficiency are irregular.

However, it is easy to control the specific saponin conversion reaction when the concentration of extracted solid is below a certain level (5%). That is, it is preferable to carry out the amount of the subcondition, which is the subject of the conversion reaction, at least 20 times the extracted solid content.

Since the subcritical water reaction temperature and the water content affect the specific saponin content, the conversion efficiency and the ease of the conversion reaction control, it is necessary to appropriately adjust the water mixing ratio according to the reaction temperature of the subcritical water.

Regarding the reaction time of the sub-critical water, it is 20 to 30 minutes at 150 ° C, A reaction time of 15 to 25 minutes at 160 DEG C and 10 to 20 minutes at 170 DEG C is sufficient but a reaction time of 0 to 10 minutes is sufficient at 180 DEG C. As the reaction temperature increases, the hydrolysis ability of the sub- As the reaction temperature increases, the reaction time must be shortened.

Step 5: Recovery of transformed reaction solution in four steps

The extracted solution after the conversion reaction may be recovered in the form of an extract after being cooled, but the extract may be concentrated into an extract by vacuum distillation, etc. Alternatively, the extract or concentrated extract may be spray dried (SD) or lyophilized FD), etc. However, since the extract liquid in the four-step reactor is in a high-temperature and high-pressure state, when the reaction is completed, the liquid is directly injected into the concentrate through the injection nozzle connected to the reactor through the injection nozzle, Is possible.

As described above, the present invention is an epoch-making method that can convert not only the total saponin content but also the specific saponin content in the total saponin content by 95% or more when the conversion reaction is performed under the optimum subcritical water condition.

The optimum condition of the subcritical water is a condition for converting the ginseng (ginseng or white ginseng) extract itself, and the ginseng (ginseng or white ginseng or red ginseng) itself is not a ginseng extract It is not a condition.

Claims (7)

The ginseng hot water extract obtained by hot water extraction of ginseng was passed through a subcritical water reactor for 15 to 25 minutes at a temperature of 150 ° C. and a pressure of 0.48 MPa for 20 to 30 minutes, at a temperature of 160 ° C. and a pressure of 0.62 MPa, or at a temperature of 170 ° C. and a pressure of 0.79 Rpa and Rd are converted into Rg3, Rk1 and Rg5 by converting Ginsenoside Rg1 and Re in Ginsenoside to Rg2, Rk3 and Rh4, and Ginsenosides Rb1, Rb2 and Rd are converted into Rg3, Rk1 and Rg5. The method according to claim 1, wherein the ginseng extract is selected from the group consisting of fresh ginseng, white ginseng, and red ginseng extract. delete delete delete delete delete

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