WO2013015504A1 - Method for deglycyrrhizinating liquorice, deglycyrrhizinated liquorice extracts and method for preparing same, and anti-caries composition - Google Patents

Abstract

The present invention relates to a method for deglycyrrhizinating liquorice, to deglycyrrhizinated liquorice extracts and to a method for preparing same, and to an anti-caries composition.

Description

Method of removing glycyrrhizine from licorice, licorice extract from glycyrrhizine, and preparation method thereof

The present invention relates to a method for removing glycyrrhizine of licorice, a licorice extract from which glycyrigin has been removed, and a method for preparing the same, and more particularly, to extracting licorice from hot water and solvent extraction, and then using the porous resin adsorbent. The present invention relates to a method for removing glycyrrhizine of licorice, which is capable of effectively removing glycyrizine in licorice, and to a method for preparing licorice extract from which glycyridin has been removed, and an anti-carbohydrate composition comprising the licorice extract from which glycisgin is removed.

As dietary patterns are diversified, the consumption of sugars is increased, increasing the number of microorganisms in the oral cavity. Tooth decay is a bacterial dental disease that causes the decay of dental tissue by demineralizing the minerals in the teeth and destroying dentin by causing bacteria in the oral cavity, especially Streptococcus mutans. It is reported to have.

In general, tooth decay occurs when three conditions exist: teeth, mutans, and carbohydrates. Mutans bacteria decompose sucrose, an energy source, to be used for growth, and produce glucose and fructose using sucrose as a substrate. Glucose produced is reported to be a major cause of forming plaque, which is an attachment group of microorganisms.

Currently, research on anti-caries has been conducted in the food-related field, and the research on separation of substances having anti-cartococcus activity from natural substances and sweetening ingredients replacing sucrose has been mainstream. For example, catechins and the like contained in green tea and oolong tea have been found to have inhibitory activity against anti-detox bacteria. In addition, licorice extract has been reported to have a strong inhibitory activity against anti-detox bacteria.

For example, Korean Patent No. 10-0454226 discloses a method for preparing licorice extract that inhibits the growth and activity of cavities and a functional anti-cavity material containing licorice extract.

However, licorice extract contains a large amount of glycyrrhizin (glycyrrhizin), which exhibits side effects in the human body, along with components that inhibit the activity of caries. In general, glycyrazine is known to have side effects such as hypertension, edema, hypokalemia, hyporeninemia and the like. Accordingly, in order to use licorice extract as a human application material for anti-cavities, glycyrrhizin should be removed. In the above patent, glycyrrhigin is removed by hydrothermal extraction. However, these methods alone cannot effectively remove glycyrrhizin in licorice.

Accordingly, an object of the present invention is to provide a method for removing glycyrrhizine of licorice which can effectively remove glycyrrhizine in licorice.

In another aspect, the present invention provides an anti-carbohydrate composition comprising a licorice extract having an excellent antimicrobial activity (anti-cariogenic performance) without the side effect caused by glycyrrhizin effectively removed, and a method for preparing the same There is a purpose.

The present invention to achieve the above object,

A first extraction step of extracting hot water from licorice;

Filtering the first extracted licorice, and then extracting the organic liquor by adding an organic solvent to the licorice residue;

An adsorption step of contacting the licorice extract obtained by the second extraction with a porous resin adsorbent; And

Provided is a method for removing glycyrrhizine of licorice comprising a desorption step of separating the licorice extract adsorbed on the porous resin adsorbent from the porous resin adsorbent.

In addition, the present invention,

A first extraction step of extracting hot water from licorice;

Filtering the first extracted licorice, and then extracting the organic liquor by adding an organic solvent to the licorice residue;

An adsorption step of contacting the licorice extract obtained by the second extraction with a porous resin adsorbent; And

It provides a licorice extract comprising a desorption step of separating the licorice extract adsorbed on the porous resin adsorbent from the porous resin adsorbent.

At this time, in the adsorption step, the dilution solvent is added to the licorice extract obtained by the second extraction and diluted, and then contacted with a porous resin adsorbent.

According to a preferred embodiment, the adsorption step includes the steps of: (a) diluting the licorice extract obtained by the second extraction with water, followed by primary contact with a porous resin adsorbent; And (b) adding 30 to 60% by weight of an ethanol aqueous solution to the first contacted licorice extract, followed by dilution, followed by a second contact with a porous resin adsorbent.

In addition, in the desorption step, licorice extract is preferably separated from the porous resin adsorbent using a dilute solvent, preferably 70 to 98% by weight of an ethanol aqueous solution.

In addition, the present invention provides a licorice extract, which is prepared according to the preparation method and whose content of glycyrrhizine is 1.0 wt% or less.

In addition, the present invention provides an anti-cavities composition comprising the licorice extract.

According to the present invention, after licorice is extracted with hot water and an organic solvent, a licorice extract from which glycyrizine is effectively removed by adsorption and separation through a porous resin adsorbent can be obtained. It is possible to obtain an anti-cavity composition that can be used without any worry.

1 is a graph showing the results of the antimicrobial evaluation of licorice extract prepared according to an embodiment of the present invention.

Figure 2 is a graph showing the growth inhibition evaluation results of Streptococcus mutans (S.mutans) bacteria in the presence of glucose (glucose) of the licorice extract prepared according to an embodiment of the present invention.

Figure 3 is a graph showing the results of growth inhibition evaluation of Streptococcus mutans (S.mutans) in the presence of sucrose (sucrose) of the licorice extract prepared according to an embodiment of the present invention.

Hereinafter, the present invention will be described in detail.

First, the glycyrrhizin removal method of licorice according to the present invention includes a first extraction step of extracting hot water from licorice; A second extraction step of filtering the mixture of licorice and extract obtained by the first extraction to separate licorice, adding an organic solvent to the separated licorice, and then heating and extracting it; An adsorption step of filtering the mixture of licorice and extract obtained by the second extraction to separate the extract and then adsorbing the separated extract with a hydrophobic porous resin adsorbent to adsorb pharmacologically active ingredients; And a desorption step of contacting the porous resin adsorbent, to which the pharmacologically active active ingredient is adsorbed, with a solvent so as to dissolve the pharmacologically active active ingredient in a solvent. According to the removal method of the present invention, glycyrrhizine is effectively removed from the licorice extract, which is the finally obtained pharmacologically active ingredient, in an amount of 1% by weight or less, preferably 0% by weight. The detailed description of each step is as follows.

First Extraction Step (Hot Water Extraction)

Licorice to be extracted in the present invention is preferably the root of licorice. At this time, the root of licorice is good to be dried, for example, it is crushed to the size of 0.2 ~ 5cm, it can be used to crushed to the size of 50㎛ ~ 1mm.

First, hot water is extracted from the above licorice (root). This hydrothermal extraction primarily removes glycyrazine along with foreign matter. Although it does not specifically limit, It is preferable to mix 10 to 30 times the weight of licorice with licorice, and to heat-extract hot water at 40-150 degreeC. At this time, when the hot water extraction temperature is less than 40 ° C, the elubility of glycyrrhizine may be insignificant, and when it exceeds 150 ° C, pharmacologically active active ingredients such as anti-cavities components may be extracted. In view of this point, the hot water extraction temperature is more preferably 60 to 120 ° C.

In addition, the hot water extraction may be performed, for example, for 10 minutes to 3 hours. If the hydrothermal extraction time is too short, less than 10 minutes, the solubility of glycyrizine may be insignificant, and if it is too long for more than 3 hours, the synergistic effect of the excess time may not be so great and undesirable in terms of energy cost.

In addition, hot water extraction is one or more times. Preferably, while proceeding within the temperature range and time range, the extraction is repeated three or more times under normal pressure in terms of the removal efficiency of glycyrazine. For example, it is good to extract 3 to 5 times at normal pressure. In addition, hot water extraction can be extracted by immersing licorice root in the container of a network structure, for example, immersing in the container containing water.

Through the above hydrothermal extraction (primary extraction), glycyrrhizin contained in the licorice (root) is primarily removed.

Second extraction step (organic solvent extraction)

After licorice is separated by filtration from the mixture of licorice and extract obtained by the first extraction, an organic solvent is added to the separated licorice, and then the mixture of the licorice and the organic solvent is heated to solvent extraction. That is, after hot water extraction, licorice and the extract (filtrate containing glycyrrhizine) are filtered through filter paper or the like, and the licorice is separated by filtration. The organic liquor is mixed with the separated licorice and then extracted again. At this time, glycyrrhizin contained in the extract may be recovered through a purification process.

The organic solvent is not limited. The organic solvent may include one or more selected from, for example, ethanol, methanol, butanol, propanol, ethyl acetate, acetone, chloroform, nucleic acid and the like, and preferably includes ethanol. As an organic solvent, 30-98 weight% of ethanol aqueous solution can be used specifically ,.

Although not specifically limited at this time, it is preferable to mix 5-30 times the organic solvent (for example, ethanol) of licorice weight, and to heat-extract the solvent at 30-100 degreeC. By solvent extraction (secondary extraction) using such an organic solvent, an extract containing pharmacologically active active ingredients such as anti-cavities components is extracted. At this time, when the organic solvent is used less than 5 times the weight of licorice, sufficient extraction may not be made. When the amount of the organic solvent used is more than 30 times, the synergistic effect of using the excess solvent is not very large, and the concentration time may be long. In addition, when the extraction temperature is less than 30 ° C, the dissolution properties of the active ingredient (anti-cause components, etc.) may be insignificant, and when the extraction temperature exceeds 100 ° C, the volatility of the organic solvent may increase. In view of this point, it is more preferable to add an organic solvent at 10-20 times the weight of licorice and extract it at a temperature of 50-90 ° C.

In addition, the said solvent extraction time is 10 minutes-4 hours, for example. If the extraction time is too short, less than 10 minutes, the elutability of the active ingredient may be insignificant, and if the extraction time is too long over 4 hours, the synergistic effect of the excess time is not so large and may be undesirable in terms of energy costs. In addition, the solvent extraction is preferably performed at least once under reduced pressure. Preferably, while proceeding within the temperature range and time range, it is good to extract at least once under a pressure of 10 ~ 700 Torr.

Through solvent extraction (secondary extraction) using the organic solvent as described above, an extract obtained by extracting pharmacologically active active ingredients such as anti-cavities components contained in licorice is obtained. The extract thus obtained is subjected to the adsorption step described below. That is, after the solvent extraction (secondary extraction) as described above, licorice (licorice foil) and the extract liquid is separated through a filter paper, etc., the separated extract is subjected to the adsorption step below. At this time, the obtained extract may be subjected to the following adsorption step after being concentrated or diluted in some cases.

Adsorption step

The extract obtained through the secondary extraction is contacted with a porous resin adsorbent. In contacting the extractant with the porous resin adsorbent, a batch type or a continuous flow type may be used. For example, the extract and the porous resin adsorbent may be put together in a vessel provided with a stirrer and contacted by a batch method. At this time, the porous resin adsorbent can be contacted by adding, for example, 10 to 100 times the weight of the extract. Preferably, as a continuous flow using a column, the porous resin adsorbent is packed into the column, and then the extract is contacted (adsorption) by continuously passing the extract through the column filled with the porous resin adsorbent. It is good to let. As described above, in the case of the continuous flow type using the column, a large amount of the extract liquid can be continuously contacted within a shorter time than the batch type, so the efficiency is good.

By contacting with the porous resin adsorbent as described above, glycyrrhizin is secondarily separated and removed, thereby obtaining a licorice extract which is a pharmacologically active ingredient in which glycyrrhizin is effectively removed. That is, the extract obtained through the solvent extraction (secondary extraction) contains a small amount of glycyrrhizin together with the pharmacologically active active ingredients such as anti-cavities components, wherein the pharmacologically active active ingredients such as the anti-cavities components included in the extract are porous The glycyrrhizine adsorbed by the resin adsorbent and remaining in the extract is separated and removed. Specifically, the porous resin adsorbent may be used to selectively adsorb a hydrophobic material. The hydrophobic pharmacologically active active ingredient is adsorbed onto the porous resin adsorbent, and the hydrophilic glycyrrhizin is separated and removed. Accordingly, the licorice extract adsorbed on the porous resin adsorbent is free or contains trace amounts of glycyrrhizin.

In the present invention, the porous resin adsorbent is a resin molded article containing a plurality of pores, which may be hydrophobic. Specifically, the porous resin adsorbent may be hydrophobic with a porous structure capable of adsorbing pharmacologically active active ingredients such as anti-cavity components without adsorbing glycyrrhizin. In addition, the shape and size of the porous resin adsorbent are not limited. Porous resin adsorbents may have, for example, the shape of bulks, pellets, or particles. In addition, the porous resin adsorbent may have a size (diameter, width, length) of, for example, 50 μm to 5 cm in the form of pellets or particles.

The resin constituting the porous resin adsorbent is not limited as long as it is hydrophobic. The porous resin adsorbent may be one obtained by molding a synthetic resin such as an aromatic or aliphatic system into a porous body. The porous resin adsorbent may include, for example, one or more selected from styrene-divinylbenzene, methacrylate, and derivatives thereof. For example, the porous resin adsorbent introduces a substituent such as a sulfone group into the benzene ring of styrene-divinylbenzene as a polymer of styrene-divinylbenzene, a polymer of methacrylate, or a derivative of styrene-divinylbenzene. As molded from the polymers, it is possible to use those having a specific surface area of 400 to 1,000 m 2 / g including a plurality of pores. In addition, the porous resin adsorbent may use a commercially available product. For example, HP series (HP10, HP20, HP30, HP40, etc.) or SP series (SP800, SP825, SP850, etc.) as styrene-divinylbenzene, and HPMG series (HP1MG, HP2MG, etc.) as methacrylates. May be used, and preferably HP20 may be usefully used as the HP series.

In addition, in contacting (adsorbing) the extract obtained through the second extraction with the porous resin adsorbent as described above, the dilution solvent is added to the extract according to the viscosity of the extract obtained through the second extraction, followed by dilution. Can be contacted with a resin adsorbent. In this case, the dilution solvent may be used at least one selected from water and an organic solvent. And the organic solvent is as listed above, preferably an ethanol aqueous solution can be used.

In contacting (adsorption) of the extract obtained through the second extraction and the porous resin adsorbent, (a) dilution by adding water to the licorice extract obtained by the second extraction, followed by the porous resin adsorbent and the first Secondary contacting process; And (b) diluting the first contacted licorice extract by diluting with a low concentration of an organic solvent, followed by a second contact with a porous resin adsorbent.

As described above, in the case where the second contact of the step (a) and the step (b) is performed continuously, the resin adsorbent increases the adsorption amount of the pharmacologically active active ingredient, and the hydrophilic glycyrrhizin is separated from the water of the step (a) and ( b) The removal rate may be improved by dissolving and removing the solution in a low concentration organic solvent solution. That is, in the case of continuous contacting (adsorption) through the steps (a) and (b), the separation and removal are performed so that the content of glycyrizine is close to zero and preferably the content of glycyrrhizine is zero. can do.

At this time, the organic solvent solution of low concentration in the step (b) is an aqueous solution of 60% by weight or less. That is, the low concentration organic solvent aqueous solution is an aqueous solution of 60% by weight of ethanol based on the total aqueous solution. When the content of ethanol exceeds 60% by weight, the pharmacologically active active ingredient adsorbed on the resin adsorbent may be dissolved and separated. The low concentration organic solvent aqueous solution is specifically 30 to 60% by weight of ethanol aqueous solution. Preferably, it is 50 weight% or less, More specifically, 30-50 weight% of ethanol aqueous solution is preferable.

Desorption Step

After the adsorption step is carried out as described above, the desorption step of separating the pharmacologically active ingredient adsorbed on the porous resin adsorbent from the porous resin adsorbent is performed. Specifically, after the adsorption step, the porous resin adsorbent is separated and removed so that the content of glycyrrhizine is close to zero (zero) or zero (zero), and the licorice extract containing a large amount of pharmacologically active active ingredients such as anti-cavities (hereinafter, ' Licorice extract 'is adsorbed, which is obtained by separating it from the porous resin adsorbent.

In the present invention, the desorption can be considered various methods if the licorice extract adsorbed on the porous resin adsorbent can be considered, for example, a dissolution method using a solvent can be selected. Specifically, the porous resin adsorbent on which the licorice extract is adsorbed may be contacted with a solvent to separate the licorice extract by dissolving it in the solvent. As the solvent, preferably 70 to 98% by weight of an ethanol aqueous solution can be used. In this case, when the concentration of ethanol is less than 70% by weight, the solubility may be low, and the yield may be insignificant. When it exceeds 98% by weight, the ethanol aqueous solution may have high volatility, which may be undesirable.

In addition, in advancing desorption through a solvent as mentioned above, a batch type may be sufficient and continuous flow type may be sufficient. Specifically, the porous resin adsorbent on which the licorice extract is adsorbed and a solvent (preferably 70 to 98% by weight of an ethanol aqueous solution) are put together and stirred (dissolved) in a vessel provided with a stirrer. Preferably it is a continuous flow using a column. Specifically, the porous resin adsorbent on which licorice extract is adsorbed is filled into the column, and then contacted (dissolved) by continuously passing a solvent (preferably 70 to 98% by weight of an ethanol aqueous solution) through the column. . In the case of continuous flow using a column, the process is more monotonous than a batch and can proceed continuously with the adsorption step. As described above, the adsorption step is a continuous flow formula using the column in terms of process efficiency, in this case, if the desorption step also uses the column, it can proceed to the method of passing the solvent through the column immediately after the adsorption, adsorption and desorption Can proceed continuously.

As described above, according to the present invention, by the adsorption through the porous resin adsorbent, the remaining glycyrrhizin in the licorice extract is secondarily removed. That is, primary separation is removed by hot water extraction, and secondary removal is performed by adsorptive separation through a porous resin adsorbent. Accordingly, according to the present invention, the licorice extract finally obtained as described above, that is, the licorice extract obtained through the adsorption step and the desorption step, the content of glycyrizine is 1.0% by weight or less, specifically 0 (zero)% by weight To 1.0% by weight. Preferably, glycyrazine is effectively removed as zero weight percent. Licorice extract, in which glycyrrhizin has been effectively removed, can be used in anti-cavity compositions as described below.

Hereinafter, a method for preparing a licorice extract according to the present invention, and a licorice extract prepared according to the present invention, and an anti-cavity composition comprising the licorice extract.

First, the method for preparing licorice extract according to the present invention includes at least four steps described in the above-described glycidazine removal method. Specifically, the first extraction step of extracting hot water licorice;

A second extraction step of filtering the mixture of licorice and extract obtained by the first extraction to separate licorice, adding an organic solvent to the separated licorice, and then heating and extracting it;

An adsorption step of filtering the mixture of licorice and extract obtained by the second extraction to separate the extract and then adsorbing the separated extract with a hydrophobic porous resin adsorbent to adsorb pharmacologically active ingredients; And

And a desorption step of separating the pharmacologically active active ingredient by dissolving it in contact with a solvent by contacting the porous resin adsorbent to which the pharmacologically active active ingredient is adsorbed. Since each of these steps is as described above, a detailed description thereof will be omitted.

According to a preferred embodiment, the method for preparing licorice extract according to the present invention is to be carried out after the three steps (the first extraction step, the second extraction step, the adsorption step and the desorption step), the licorice extract Concentrating may be further included. Specifically, when the desorption step is carried out by a method of contacting (dissolving) a solvent, the licorice extract contains a solvent (ethanol aqueous solution), in which case it is heated to a temperature of 30 ~ 100 ℃ according to the use of licorice extract Concentrating step (concentration step) may further include. At this time, when the heating temperature at the time of concentration is less than 30 ℃ low volatility of the solvent may take a long time to concentrate, if it exceeds 100 ℃ may also be volatile pharmacologically active active ingredients such as anti-cavities components.

The concentration step is preferably carried out under reduced pressure, for example, it is preferably carried out under a pressure of 10 ~ 700 Torr within the temperature range. Concentration in this pressure range is advantageous in terms of concentration efficiency. The heating time in the concentration step is not particularly limited, but may be 10 minutes to 8 hours within the temperature range. In this case, when the heating time is less than 10 minutes, the concentration efficiency is lowered, and when more than 8 hours, the synergistic effect according to the excess time is not so large and may be undesirable in terms of energy cost.

In addition, the extract concentrated through the concentration step as described above, may be dried through natural drying, hot air drying or freeze drying, if necessary. The dried extract may be commercialized in a powdered state or mixed in various formulation compositions, such as anti-cavity compositions.

In addition, the method for preparing licorice extract according to the present invention may further comprise adding an excipient to the concentrated extract. At this time, the excipient imparts water solubility to the licorice extract, or acts as a viscosity regulator for concentration, it may be selected from water-soluble sugars or fatty acids. Excipients can be used, for example, one or more selected from carrageenan, lactose, dextrin, cyclodextrin, casein, pectin, xanthan gum and pectin and the like. Such excipients may be added, for example, from 1 to 100 parts by weight based on 100 parts by weight of the concentrated extract. At this time, when the addition amount of the excipient is less than 1 part by weight, water solubility provision or viscosity control ability (concentration) may be insignificant according to the addition thereof. And when the addition amount of the excipient exceeds 100 parts by weight, the content of the licorice extract is relatively low, the anti-cavity improvement effect according to the use of licorice extract may be insignificant.

In addition, the licorice extract to which the excipient is added as described above, may be dried through natural drying, hot air drying or freeze drying, as needed. The dried extract may be commercialized in a powdered state or mixed in various formulation compositions, such as anti-cavity compositions.

On the other hand, the licorice extract according to the present invention is a glycyrrhigin is removed, prepared according to the method for producing a licorice extract according to the present invention as described above, and contains no or very low content of glycyrrhizin. Specifically, the licorice extract according to the present invention is prepared by the same method as described above, and includes pharmacologically active ingredients such as anti-cavities as an active ingredient, but contains less than 1.0% by weight of glycidazine. Preferably, glycyrrhizin is effectively removed so that the content of glycyrrhizine is zero weight percent.

In addition, the formulation of the licorice extract according to the present invention is not limited. Licorice extract according to the present invention may be selected from, for example, liquid, paste, powder (solid) and the like. For example, after concentration as described above, it may have a powdery formulation through drying.

Licorice extract according to the present invention is effectively removed as described above glycerin, there is no side effect due to glycyrrhizin. In addition, the licorice extract according to the present invention is in the presence of sugars, for example, in the presence of sugars such as glucose (glucose) or sucrose (sucrose) of caries-inducing bacteria such as Streptococcus mutans (S.mutans) bacteria It effectively suppresses growth. Accordingly, the licorice extract according to the present invention can be usefully applied (included) as an anti-carcinoma active ingredient for inhibiting dental caries in foods such as confectionary, candy, candy, chocolate and gum containing a lot of sugar components. have.

In addition, the anti-cavities composition according to the present invention, the licorice extract of the present invention. The formulation of the anti-cavities composition in the present invention is not limited. The anti-cavities composition according to the present invention, including the licorice extract of the present invention, is not limited as long as it has anti-cavities performance (antibacterial), for example, it can be selected from pharmacological compositions, food compositions and oral compositions and the like. have. For example, the food composition may include sweets, candy, candy, chocolate, gum composition, and the like. The oral composition may include, for example, a toothpaste composition and an oral cleaning composition. For example, when the anti-cavities composition according to the present invention is a toothpaste composition, the anti-cavities composition according to the present invention comprises a toothpaste base component and the licorice extract of the present invention. In this case, the toothpaste base component constitutes a conventional toothpaste composition, which may include, for example, an abrasive and a surfactant.

In addition, the anti-cavity composition according to the present invention, the total licorice extract may comprise 1.0 ppm to 10.0% by weight based on the total weight of the composition. At this time, if the content of licorice extract is less than 1.0ppm, the effect such as anti-cavities according to the content of licorice extract may be insignificant, and if the content exceeds 10.0% by weight, the anti-cariogenic function difference according to the increase of the content of licorice extract is not large May affect the formulation of the drug. In consideration of this point, the licorice extract is preferably included in 30 ~ 100ppm based on the total weight of the anti-cavities composition.

According to the present invention as described above, after hydrothermal extraction and organic solvent extraction, glycyrrhizin in licorice is effectively removed by adsorptive separation through a porous resin adsorbent. Accordingly, it is possible to provide a licorice extract having no side effects.

Hereinafter, the Example of this invention is illustrated. The following examples are merely provided to aid the understanding of the present invention, whereby the technical scope of the present invention is not limited.

EXAMPLE

Hot Water and Solvent Extraction

1 g of water was mixed with 50 g of dried licorice root, and extracted with hot water three times at a temperature of 90 ° C. for 2 hours. Then, the licorice extracted by hot water was filtered, the extract was separated and removed, and an aqueous solution of 75 wt% ethanol having 15 times the weight of licorice was added to the remaining licorice after filtration and extracted for 2 hours at a reduced pressure and a temperature of 85 ° C.

* <Adsorption and desorption>

Next, 6 L of water was added to the extract obtained by extraction with the ethanol aqueous solution, followed by mixing and diluting. Then, the diluted extract was passed through a column filled with a porous resin adsorbent for primary contact. At this time, the column was used as a porous resin adsorbent filled with HP20 (Japan, Mitsubishi Chemical can use).

As above, after obtaining the first contacted extract, 2,600 ml of a 50% by weight aqueous solution of ethanol was added thereto, mixed, and passed through the column again to make a second contact.

Subsequently, 2,500 ml of a 95 wt% ethanol aqueous solution was passed through the column to dissolve licorice extract, a pharmacologically active ingredient adsorbed on the porous resin adsorbent.

<Concentration and drying>

Then, a licorice extract obtained through the column was obtained, and it was concentrated under reduced pressure for 8 hours at a temperature of 85 ° C. Thereafter, carrageenan (10 wt% total weight of the concentrated extract) was added to the concentrated licorice extract as an excipient and concentrated to obtain a paste extract. Next, lyophilized for 3 days at -50 ℃, 15 Torr to prepare a licorice extract in powder form.

Comparative Example 1

Compared with the above examples, the same procedure was followed except that hot water extraction, adsorption and desorption were not performed. Specifically, licorice was extracted without hot water extraction. Thereafter, the mixture was immediately concentrated without contacting the resin adsorbent (HP20), and then an excipient (carrageenan) was added, followed by freeze drying, which was used as a licorice extract specimen according to Comparative Example 1.

Comparative Example 2

Compared with the above examples, the same procedure was followed except that adsorption and desorption were not performed. Specifically, licorice was concentrated immediately after performing hot water extraction and ethanol extraction, without contacting the resin adsorbent (HP20). After concentration, an excipient (carrageenan) was added, and then lyophilized was used as a licorice extract specimen according to Comparative Example 2.

For licorice extracts prepared according to the above Examples and Comparative Examples, the content of glycyrrhizin was measured using HPLC (High-Pressure Liquid Chromatography), and the results are shown in the following [Table 1]. The content (% by weight) of the following [Table 1] is based on the total weight of licorice extract.

Table 1 <Glycirizine Contents of Licorice Extracts>

division	Glycyrrhizin content (% by weight)
Example	0 (not detected)
Comparative Example 1	4.8645
Comparative Example 2	2.2056

As shown in Table 1 above, when only ethanol extraction was carried out as in Comparative Example 1, it can be seen that it contains a high content of glycyrrhizin as 4% by weight or more. And in the case of Comparative Example 2 subjected to hot water extraction and ethanol extraction continuously, it can be seen that the content of glycyrizine is less than 2% by weight, but is not effectively removed. However, after contact with the porous resin adsorbent (HP20) after hot water and ethanol extraction according to an embodiment of the present invention, it can be seen that the content of glycyrrhizin is effectively removed as 0 (not detected).

In addition, Figure 1 is a graph showing the results of the antimicrobial evaluation according to the concentration of licorice extract prepared according to the embodiment, the graph showing the MIC (Minimum Inhibitory Concentration) results as O.D (Optical Density) value. At this time, the antimicrobial evaluation results were performed using the Streptococcus mutans (S.mutans), a decay-inducing bacterium, as the MIC (Minimum Inhibitory Concentration) test method recommended by the National Committee of Clinical Laboratory Standard (NCCLS) as follows. .

1. Dissolve licorice extract in ethanol to a concentration of 10mg / ml

2. Dilute 1 / 10-fold with autoclaved distilled water to a concentration of 1mg / ml

3. Dilute to desired concentration with Triptic Soy Broth (TSB) medium (30ppm, 40ppm, 50ppm, 60ppm, 70ppm, 80ppm, 90ppm and 100ppm)

4. Dispense the diluted licorice extract into the 96-well plates in 150 µl steps.

5. Streptococcus mutans cultured in anaerobic conditions at 37 ℃ in TSB medium

6. Dispense 150 μl of medium containing Streptococcus mutans at a concentration of 10 ⁶ colony-forming units / ml into a 96-well plate (diluted licorice extract).

7. Immediately measure O.D values and measure O.D one more time before incubation

8. Incubate in anaerobic condition at 37 ℃ and measure O.D value every 16 hours, 24 hours, 40 hours to obtain graph

As shown in FIG. 1, the licorice extract according to the present invention was confirmed to have antimicrobial activity against the Streptococcus mutans (S.mutans). In addition, the antimicrobial (O.D value) was found to increase as the concentration of licorice extract increases. In addition, as shown in the graph of Figure 1, the licorice extract according to the embodiment of the present invention was found to have excellent antimicrobial activity by inhibiting the activity of Streptococcus mutans (S.mutans) even at low concentrations of 30ppm and 40ppm .

On the other hand, with respect to the licorice extract prepared according to the above embodiment, in order to determine the growth inhibitory effect of Streptococcus mutans (S.mutans) bacteria in the presence of glucose (glucose) and sucrose (sucrose), biofilm ( Biofilm) was used to measure the OD value. The results are shown in FIGS. 2 and 3. Figure 2 is a graph showing the measurement results performed in the presence of glucose (glucose), Figure 3 is a graph showing the measurement results performed in the presence of sucrose (sucrose).

Biofilm used a 24-well polystyrene (flat-bottom) cell culture cluster (Costar 3596; manufactured by Corning). The biofilms were grown to Streptococcus mutans (S.mutans) to OD ₆₀₀ = 0.5.

First, to determine the growth inhibitory effect of Streptococcus mutans (S.mutans) in the presence of glucose (glucose), 2mL saliva (Saliva) of licorice extract prepared according to the above example at each concentration (10 ~ 100ppm) ) And 20 mM glucose was added thereto. In the graph of FIG. 2, 'Saliva coating' is a result of culturing bacteria (S.mutans) after coating saliva (Saliva) containing licorice extract on a biofilm. In the graph of FIG. 2, 'Saliva adding' is a result of incubating saliva (Saliva) and bacteria (S.mutans) together with licorice extract. At this time, the culture of bacteria (S.mutans) was carried out for 48 hours in 37 ℃, 5wt% CO ₂ environment. In addition, the licorice extract was compared to the untreated specimens ('No saliva treatment' in the graph of Figure 2).

As shown in Figure 2, the licorice extract prepared according to the embodiment was found to have a growth inhibitory effect on the Streptococcus mutans (S.mutans) bacteria in the presence of glucose (glucose). In particular, it can be seen that it has a growth inhibitory effect even at a low concentration of 20ppm.

In addition, in order to determine the growth inhibitory effect of S. mutans bacteria in the presence of sucrose, the OD value was evaluated in the same manner as above and the results are shown graphically in FIG. It was. In the graph of FIG. 3, 'Saliva coating' is the result of culturing bacteria (S.mutans) after coating, 'Saliva adding' is the result of culturing with bacteria (S.mutans). 'No saliva treatment' is the result of untreated specimens without licorice extract.

As shown in FIG. 3, the licorice extract prepared according to the embodiment was found to have a growth inhibitory effect of Streptococcus mutans bacteria in the presence of sucrose. In case of coating, good results were obtained even at low concentrations, and in the case of adding, good results were obtained at concentrations of more than 50 ppm of licorice extract.

As confirmed through the above experimental example, licorice extract prepared according to the present invention is effective in the growth of Streptococcus mutans (S.mutans) even in the presence of sugars such as glucose (glucose) or sucrose (sucrose) It can be seen that. Accordingly, it can be seen that it can be usefully applied to foods with a high content of sugar such as sweets, candy, chocolate, and the like.

Claims (16)

1. A first extraction step of extracting hot water from licorice;

   A second extraction step of filtering the mixture of licorice and extract obtained by the first extraction to separate licorice, adding an organic solvent to the separated licorice, and then heating and extracting it;

   An adsorption step of filtering the mixture of licorice and extract obtained by the second extraction to separate the extract and then adsorbing the separated extract with a hydrophobic porous resin adsorbent to adsorb pharmacologically active ingredients; And

   A desorption step of separating the pharmacologically active active ingredient is dissolved in a solvent by contacting the porous resin adsorbent adsorbed to the pharmacologically active active ingredient so as to dissolve in the solvent.
2. The method of claim 1,

   Said adsorption step, diluting by adding a dilution solvent to the extract separated in the second extraction, and then contacting with a porous resin adsorbent, licorice glycidazine removal method.
3. The method of claim 1,

   The adsorption step is

   (a) adding water to and diluting the extract separated in the second extraction, followed by first contact with a porous resin adsorbent; And

   (b) adding a dilute solution of 30 to 60% by weight of ethanol to the first contacted extract, followed by a second contact with a porous resin adsorbent for licorice glycidazine removal.
4. The method according to any one of claims 1 to 3,

   The adsorption step, Glycyrrhizine removal method of licorice, characterized in that the extract is passed through a column filled with a porous resin adsorbent.
5. The method according to any one of claims 1 to 3,

   The desorption step, Glycirizine removal method of licorice, characterized in that to separate the pharmacologically active ingredient from the porous resin adsorbent using 70 ~ 98% by weight of ethanol aqueous solution.
6. The method according to any one of claims 1 to 3,

   The porous resin adsorbent is glycerine-divinylbenzene, methacrylate and derivatives thereof, characterized in that at least one selected from the group consisting of glycyrrhizin of licorice.
7. A first extraction step of extracting hot water from licorice;

   A second extraction step of filtering the mixture of licorice and extract obtained by the first extraction to separate licorice, adding an organic solvent to the separated licorice, and then heating and extracting it;

   An adsorption step of filtering the mixture of licorice and extract obtained by the second extraction to separate the extract and then adsorbing the separated extract with a hydrophobic porous resin adsorbent to adsorb pharmacologically active ingredients; And

   And a desorption step of separating the pharmacologically active active ingredient by dissolving the pharmacologically active active ingredient in contact with a solvent by contacting the porous resin adsorbent to which the pharmacologically active active ingredient is adsorbed.
8. The method of claim 7, wherein

   The adsorption step, dilution by adding a dilution solvent to the extract separated in the second extraction, and then contacting with a porous resin adsorbent manufacturing method of licorice extract.
9. The method of claim 7, wherein

   The adsorption step is

   (a) adding water to and diluting the extract separated in the second extraction, followed by first contact with a porous resin adsorbent; And

   (b) adding a dilute solution of 30 to 60% by weight of ethanol to the first contacted extract, followed by a second contact with a porous resin adsorbent for producing a licorice extract.
10. The method of claim 7, wherein

    The adsorption step, the licorice extract production method characterized in that the extract is passed through a column filled with a porous resin adsorbent.
11. The method of claim 7, wherein

    The desorption step, the licorice extract manufacturing method characterized in that to separate the pharmacologically active ingredient from the porous resin adsorbent using 70 ~ 98% by weight of ethanol aqueous solution.
12. The method of claim 11,

    Method of producing a licorice extract, characterized in that further comprising the step of condensing the extract after the desorption step to a temperature of 30 ~ 100 ℃ under reduced pressure.
13. The method of claim 11,

    Concentrating the licorice extract after the desorption step by heating to a temperature of 30 ~ 100 ℃ under reduced pressure; And

    Method for producing a licorice extract, characterized in that it further comprises the step of adding an excipient to the concentrated extract.
14. The method of claim 7, wherein

    The porous resin adsorbent is a method for producing licorice extract, characterized in that it comprises one or more selected from styrene-divinylbenzene, methacrylate and derivatives thereof.
15. The licorice extract is prepared by the method according to any one of claims 7 to 14, characterized in that the content of glycyrizine is 1.0% by weight or less.
16. An anti-cavities composition comprising licorice extract according to claim 15.

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