WO2015108323A1

WO2015108323A1 - Preparation method for artemisia sp. extract with reduced harmful substance content for treating stomach and intestinal disease

Info

Publication number: WO2015108323A1
Application number: PCT/KR2015/000360
Authority: WO
Inventors: 김순회; 손미원; 전준호; 박형근; 한상덕; 최상진; 차봉진; 이전평; 최나영; 홍성민; 조성열
Original assignee: 동아에스티 주식회사
Priority date: 2014-01-15
Filing date: 2015-01-14
Publication date: 2015-07-23
Also published as: KR20150085330A

Abstract

The present invention relates to a preparation method for an extract of Artemisia sp., extracted from leaves of Artemisia sp. with ethanol or propanol, for treating stomach and intestinal disease, which reduces a harmful substance content by using activated carbon. The preparation method for an extract of Artemisia sp. according to the present invention uses activated carbon, thereby reducing benzo[a]pyrene derived from a raw herb medicine so as to produce the extract of Artemisia sp. with enhanced safety and without changes in medicinal effect. Therefore, the extract of Artemisia sp. prepared by the preparation method according to the present invention can effectively reduce benzo[a]pyrene, which is the harmful substance, while retaining a medicinal effect.

Description

Method for preparing leaf extracts for the treatment of gastrointestinal diseases with reduced content of harmful substances

The present invention relates to a method for preparing a leaf extract, which can maintain the efficacy of treating gastrointestinal diseases while reducing the content of benzopyrene, a harmful substance in leaf extract.

Benzo [a] pyrene is a yellow crystalline solid belonging to the group of polyaromatic hydrocarbons (PAHs), and since it is produced by incomplete combustion at temperatures between 300 and 600 ° C., there are a variety of pollutants.

Mainly present in coal tar, automobile exhaust gas (especially diesel engine), tobacco smoke, and also in non-cooked / processed foods such as agricultural products and fisheries due to environmental pollution, and carbohydrates and proteins, which are the main components of foods at high temperature heating cooking / processing It is also produced by the decomposition of lipids.

Benzopyrene is more problematic due to its long retention period and its high toxicity.It is an endocrine barrier and carcinogen, which is included in the priority list for risk assessment of CODEX (International Food Standards Commission) and JECFA (FAOWHO Joint Food Additives Expert Committee). It has become a target material of global interest. The International Cancer Institute (IARC) recently upgraded benzopyrene to Group 1 human carcinogen. In foods, there is a problem with benzopyrene. Recently, the detection of benzopyrene in edible oils such as olive oil has become a social problem in Korea.

Regulations on benzopyrene produced during the edible oil and fat manufacturing process are set at 2.0ppb, Spain 5.0ppb, and China 10ppb, and the recommended standard is set at 2.0ppb or less in accordance with EU standards, which are strictly regulated in Korea. have.

In order to reduce such harmful benzopyrene, various biological, physical and chemical methods have been disclosed. In particular, a method of reducing benzopyrene produced in edible oils in foods is provided by a refining exhaust facility and adjusting the roasting temperature of compressed oil. It is disclosed in Korean Unexamined Patent Publication No. 2009-38548.

As a method for reducing benzopyrene in conventional edible oils, a method of absorbing edible oils into clay and carbonizing the edible oils and activating the carbon to use acid-activated clay as an absorbent is disclosed in US Patent No. 5,218,132. Is disclosed. Carbonation and activation step of the edible oil and fat in the US patent is made by applying heat in the presence of the active factor zinc chloride (zinc chloride) and the activation temperature should be 250 ℃ or more. However, the method can be applied in general edible oil, but in the case of compressed oil, especially pepper seed oil, sesame oil, perilla oil may cause a problem that the flavor and color change by clay or temperature.

Thus, a method for producing compressed benzopyrene-reduced compressed oil by using activated carbon in edible oil and fat is known (Korean Patent Application No. 2007-103889), but the prior art is made at a high temperature of 60 to 110 ℃. .

However, when using the activated carbon reduction process used in such a conventional food is required because the pretreatment process at a high temperature is not a suitable method for use in the herbal extracts because there is a problem in the change in the active ingredient.

In general, most herbal medicines are known to contain a certain amount of harmful benzopyrene due to problems such as modern environmental pollution (Monitoring on the content of benzopyrene in traditional herbal medicines, Digital Policy Research, Vol. 10, No. 7 (2012.8)). . In order to manage benzopyrenes generated during the pretreatment of herbal medicines, the Korea Food and Drug Administration sets and manages standards (5ppb) for benzopyrene, a hazardous substance, for sucrose and turmeric. -13).

The leaf extract is used as a gastritis remedy for gastric mucosa regeneration by promoting prostaglandin production, HSP, VEGF and EGF production, promoting mucus secretion and gastric mucosa, protecting gastric mucosa and promoting healing of damaged gastric cells. , Korea Patent Publication No. 10-1000951, Republic of Korea Patent Publication No. 2009-0088343 and the like is known to be extracted and manufactured using a solvent belonging to ICH Guideline Class 3, such as alcohol and isopropyl alcohol.

These leaf extracts are enriched with raw leaf herbals, benzopyrene, which is a harmful substance among leaf extracts, has affinity for organic solvents such as ethanol and isopropyl alcohol, which are used for extracting active ingredients such as eufatlin and zodiacine. High in some contaminated herbal medicines Benzopyrene may be included as the leaf extract.

Benzopyrene, which may be present in trace amounts in leaf extracts, is very low in daily intake, and the amount exposed to the human body is much smaller than when ingested with other Chinese medicines or foods. There is a potential for unsupported risk. Therefore, the removal of benzopyrene in the leaf extract is an important task to be accomplished by those skilled in the art.

Conventionally, although it is known to use activated carbon in herbal medicines, such as Korean Patent Publication No. 10-0577396, Korean Patent Publication No. 2009-0080459, etc., it is used for the purpose of removing chlorine, chloroform, and the like, which are filtration purposes in a general process. It is only one level, and there is no particular description about selectively reducing only harmful substances such as benzopyrene and minimizing the change in the content of the active ingredient.

Therefore, there is no known method for reducing benzopyrene in the leaf extract until now, and when the method used in food (Korean Patent Publication No. 2009-38548) is used, the active ingredient in the leaf extract is denatured by heat to decrease the content. And there is a problem that can show the effect on the drug.

Active carbon is a special carbon that is mainly activated by using coconut shell, wood, and coal as a raw material, and is a collection of amorphous carbon with fine pore size of micropore in the process of activation. Raw materials for activated carbon can be roughly classified into vegetable, animal, mineral and industrial waste, and minerals can be classified into coal and petroleum. Generally, vegetable raw materials are used for the production of powdered activated carbon, and charcoal, coconut shell, coal, and the like are used for producing granular activated carbon.

Adsorption theory of activated carbon refers to a phenomenon in which certain components of gas or liquid are concentrated at the solid-liquid, gas-liquid, and liquid-liquid interface. Adsorption is usually classified into physical adsorption and chemical adsorption according to the type of adsorption. The force that dominates the physical adsorption is the van der Waals force, and the force that dominates the chemical adsorption is the chemical bonding force such as ionic bond or covalent bond. In the case of physical adsorption, solutes adhere to the adsorbent surface when the attraction of molecules between the solute and the adsorbent is greater than the attraction between the solute and the solvent.

While the present inventors are conducting research to lower the content of benzopyrene derived from raw materials, etc. in the method of manufacturing the leaf extract, surprisingly, the pharmacopeia management standard of the Ministry of Food and Drug Safety of benzopyrene, which is a toxic substance while maintaining pharmacological activity (Notified by the Korea Food and Drug Administration, Bulletin 2009) -13) to complete the present invention by developing a method that can be reduced to a level suitable for 30%)

The present invention overcomes the shortcomings of the conventional manufacturing method in which the benzopyrene derived from the crude drug is converted to the extract during the preparation of the lobar extract, and selectively maintains only the content of the harmful substance benzopyrene while maintaining the contents of the active ingredients eupatillin and zodiacine. It is an object of the present invention to provide a method for producing a leaf extract, which shows a gastrointestinal disease treatment effect equivalent to that of the extract when it is not treated with activated carbon.

The present invention relates to a method for producing a leafy leaf extract for treating gastrointestinal diseases in which the leaflet leaf extract of leaflet leaves with ethanol or propanol is reduced in the amount of harmful substances using activated carbon.

In order to achieve the above object, the leaf extract according to the present invention should have the following characteristics.

First, the effect of larvae extract on gastrointestinal diseases should be maintained to an equal level.

Second, the content of benzopyrene, a harmful substance derived from the original herbal medicine, should be sufficiently reduced.

The present invention uses activated carbon to reduce the content of benzopyrene derived from the leaf of the leaf, leaf extracts treated with activated carbon with 0.1 to 20% (w / w) relative to the weight of leaf of the leaf, while significantly reducing the content of benzopyrene, eupatini, There is no significant change in the content of postural osdine, which shows the effect of treating the gastrointestinal diseases of the leaf extracts of the same level as the untreated activated carbon.

In the present invention, when using 0.1% (w / w) activated carbon relative to the weight of the leaf of the leaf, it was found that the content of benzopyrene, which is a harmful substance, is reduced by about 80% (w / w), and the content of benzopyrene remaining in the leaf extract. This trace amount was found to have no effect on the human body, and compared to the untreated carbon group, the contents of eupatillin and postiosidine were almost unchanged, and the effect of treating gastrointestinal diseases was found to be equal.

In addition, in the present invention, when using 20% (w / w) activated carbon compared to the weight of the leaf of the leaf, the content of benzopyrene as a harmful substance is reduced by about 99% (w / w) so that almost no benzopyrene remains in the leaf extract, Compared with unactivated charcoal, eufattylin and postiosidine were slightly decreased, but the gastrointestinal disease treatment effect was found to be equivalent.

By the way, leaf extract using 30 to 100% (w / w) activated carbon compared to the weight of leaf leaves can remove all of the harmful benzopyrene, but the active ingredient eupatillin, postiosidine also shows a large content change in the present invention Excluded from the category of.

Therefore, the present invention is characterized by using 0.1 to 20% (w / w) activated carbon relative to the weight of the leaf of the leaf, preferably from 0.1 to 5% (w / w) activated carbon.

Leaf leaf in the present invention can be extracted with ethanol or propanol, ethanol is not changed in the benzopyrene content reduction ability in the range of 70 ~ 100% (v / v) concentration when activated carbon filtration process, 95% (v / v) Ethanol is preferred. Propanol also uses 1-propanol or 2-propanol with a 100% (v / v) concentration.

In another aspect, the present invention provides a method for reducing the benzopyrene content by using 0.1 ~ 20% (w / w) activated carbon compared to the weight of the leaf of the leaf leaf extract ethanol or propanol.

In the present invention, the process of reducing benzopyrene in the leaf extract can be treated during the extraction of the leaf of the leaf, the treatment before the extraction of the leaf of the leaf, or the leaf of the leaf extract can be treated after re-dissolution.

In the present invention, the step of processing during extraction of the leaf of the leaf is to extract the leaf of the leaf with a solvent to remove the benzopyrene by circulating filtration using a column filled with activated carbon, the process of extracting the leaf of the leaf before the leaf is leaf After filtration and solvent extraction, the column packed with activated carbon was filtered to remove benzopyrene, and the treatment process after concentration and re-dissolution of the leaf extract was carried out by solvent extraction of the leaf of the leaf, and the extract obtained by concentration under reduced pressure was extracted. It is a method of removing benzopyrene by circulating filtration using a column filled with activated carbon after redissolution with a solvent.

In the above benzopyrene reduction method, the benzopyrene reduction rate and the amount of eupatylin and oseosidine, which are active ingredients of the leaves, can be similarly obtained in all solvents.

The present invention overcomes the shortcomings of the conventional manufacturing method in which the benzopyrene derived from the original drug is transferred to the extract during the preparation of the leaf extract, the benzopyrene content is reduced and the content of the active substance is maintained to provide a method for producing the leaf extract having medicinal effects .

1 is a graph showing the content of benzopyrene, eupatillin, and postiosidine compared to untreated activated carbon of the leaf extract.

Figure 2 is a graph showing the inhibition of gastric lesions of the leaf extract according to the activated carbon filter process compared to the activated carbon untreated leaf extract.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited by the following examples.

<실시예 1> <Example 1> 원생약 대비 활성탄 20%(w/w) 처리 95%(v/v) 에탄올 추출물의 제조Preparation of 95% (v / v) Ethanol Extracts Treated with 20% (w / w) of Activated Carbon

100 g of fine leaf leaves were cooled to 800 ml of 95% (v / v) ethanol at room temperature for 20 hours, and circulated by filtration using a column packed with 20 g of activated carbon, and then the residue was again 800 ml of 95% (v / v) ethanol. After 4 hours of cooling, the column filled with 20 g of activated carbon was filtered through a column. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<실시예 2><Example 2> 원생약 대비 활성탄 10%(w/w) 처리 95%(v/v) 에탄올 추출물의 제조Preparation of 95% (v / v) Ethanol Extracts Treated with 10% (w / w) Activated Carbon

100 g of fine leaf leaves were cooled to 800 ml of 95% (v / v) ethanol at room temperature for 20 hours, and circulated by filtration using a column filled with 10 g of activated carbon, and then the residue was again 800 ml of 95% (v / v) ethanol. After 4 hours of cooling, the column packed with 10 g of activated carbon was filtered through circulation. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<실시예 3> <Example 3> 원생약 대비 활성탄 5%(w/w) 처리 95%(v/v) 에탄올 추출물의 제조Preparation of 95% (v / v) Ethanol Extracts Treated with Activated Carbon 5% (w / w)

100 g of fine leaf leaves were immersed in 800 ml of 95% (v / v) ethanol at room temperature for 20 hours, and circulated by filtration using a column filled with 5 g of activated carbon, and then the residue was again 800 ml of 95% (v / v) ethanol. After 4 hours of cooling, the column filled with 5 g of activated carbon was filtered through a column. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<실시예 4> <Example 4> 원생약 대비 활성탄 1%(w/w) 처리 95%(v/v) 에탄올 추출물의 제조Preparation of 95% (v / v) Ethanol Extract Treated with Activated Carbon 1% (w / w)

100 g of fine leaf leaves were cooled to 800 ml of 95% (v / v) ethanol at room temperature for 20 hours, and circulated by filtration using a column filled with 1 g of activated carbon, and then the residue was again 800 ml of 95% (v / v) ethanol. After 4 hours of cooling, the column filled with 1 g of activated carbon was filtered through a column. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<실시예 5> Example 5 원생약 대비 활성탄 0.1%(w/w) 처리 95%(v/v) 에탄올 추출물의 제조Preparation of 95% (v / v) Ethanol Extracts Treated with 0.1% (w / w) of Activated Carbon

100 g of fine leaf leaves were immersed in 800 ml of 95% (v / v) ethanol at room temperature for 20 hours, and circulated by filtration using a column filled with 0.1 g of activated carbon, followed by 95% (v / v) ethanol. 800 ml was added, the mixture was cooled for 4 hours, and the column filled with 0.1 g of activated carbon was filtered through a column. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<실시예 6> <Example 6> 추출 완료후 원생약 대비 활성탄 5%(w/w) 처리 95%(v/v) 에탄올 추출물의 제조Preparation of 95% (v / v) Ethanol Extract of Activated Carbon 5% (w / w)

100 g of fine leaf leaves were extracted by cooling with 800 ml of 95% (v / v) ethanol at room temperature for 20 hours, filtered, and then cooled again by adding about 800 ml of 95% (v / v) ethanol to the residue for 4 hours. . The combined filtrates were circulated through a column filled with 5 g of activated charcoal, and concentrated under reduced pressure to obtain about 5 g of soft lead.

<실시예 7> <Example 7> 재용해 후 원생약 대비 활성탄 5%(w/w) 처리 95%(v/v) 에탄올 추출물의 제조Preparation of 95% (v / v) Ethanol Extracts Treated with Activated Carbon 5% (w / w)

100 g of fine leaf leaves were extracted by cooling with 800 ml of 95% (w / w) ethanol at room temperature for 20 hours, and filtered. Then, 800 ml of 95% (w / w) ethanol was added to the residue. . The combined filtrates were concentrated under reduced pressure to obtain an extract of about 5 g. The extract was redissolved in 1 L of 95% (v / v) ethanol, and then circulated under filtration using a column filled with 5 g of activated carbon, and concentrated under reduced pressure to obtain about 5 g of a crude extract.

The following examples extract the leaflets with 70% (v / v), 80% (v / v), 85 (v / v), 90% (v / v) and 100% (v / v) ethanol. Using the active carbon 5% (w / w) compared to the crude drug as in Example 3 to prepare a leaf extract according to the present invention.

<실시예 8> <Example 8> 70%(v/v) 에탄올 추출물-활성탄 처리-의 제조Preparation of 70% (v / v) Ethanol Extract-Activated Carbon Treatment-

100 g of fine leaf leaves were immersed in 800 ml of 70% (v / v) ethanol at room temperature for 20 hours, and circulated by filtration using a column filled with 5 g of activated carbon, and then the residue was added to 800 ml of 70% (v / v) ethanol. After 4 hours of cooling, the column filled with 5 g of activated carbon was filtered through a column. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<실시예 9> Example 9 80%(v/v) 에탄올 추출물-활성탄 처리-의 제조Preparation of 80% (v / v) Ethanol Extract-Activated Carbon Treatment-

100 g of fine leaf leaves were quenched with 800 ml of 80% (v / v) ethanol at room temperature for 20 hours, and circulated by filtration using a column filled with 5 g of activated carbon, and then the residue was again 800 ml of 80% (v / v) ethanol. After 4 hours of cooling, the column filled with 5 g of activated carbon was filtered through a column. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

Example 10 Preparation of 85% (v / v) Ethanol Extract—Activated Carbon Treatment—

100 g of fine leaf leaves were immersed in 800 ml of 85% (v / v) ethanol at room temperature for 20 hours, and circulated by filtration using a column filled with 5 g of activated carbon, and the residue was again 800 ml of 85% (v / v) ethanol. After 4 hours of cooling, the column filled with 5 g of activated carbon was filtered through a column. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<실시예 11> <Example 11> 90%(v/v) 에탄올 추출물-활성탄 처리-의 제조Preparation of 90% (v / v) Ethanol Extract-Activated Carbon Treatment-

100 g of fine leaf leaves were immersed in 800 ml of 90% (v / v) ethanol at room temperature for 20 hours, and circulated by filtration using a column filled with 5 g of activated carbon, and then the residue was again 800 ml of 90% (v / v) ethanol. After 4 hours of cooling, the column filled with 5 g of activated carbon was filtered through a column. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<실시예 12> <Example 12> 100%(v/v) 에탄올 추출물-활성탄 처리-의 제조Preparation of 100% (v / v) Ethanol Extract-Activated Carbon Treatment-

100 g of fine leaf leaves were immersed in 800 ml of 100% (v / v) ethanol at room temperature for 20 hours, and circulated by filtration using a column filled with 5 g of activated carbon, and then the residue was again 800 ml of 100% (v / v) ethanol. After 4 hours of cooling, the column filled with 5 g of activated carbon was filtered through a column. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

The following example extracts leaflets with 100% (v / v) 1-propanol and 100% (v / v) 2-propanol instead of extractant ethanol and activated carbon 5% (w) / w) to prepare a leaf extract according to the present invention.

<실시예 13> Example 13 100%(v/v) 1-프로판올 추출물-활성탄 처리-의 제조Preparation of 100% (v / v) 1-propanol Extract-Activated Carbon Treatment-

100 g of fine leaf leaves were immersed in 800 ml of 100% (v / v) 1-propanol at room temperature for 20 hours, and circulated by filtration using a column filled with 5 g of activated carbon, followed by 100% (v / v) 1 of the residue. Add 800 ml of propanol, cool for 4 hours, and filter the column filled with 5 g of activated carbon. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<실시예 14> <Example 14> 100%(v/v) 2-프로판올 추출물-활성탄 처리-의 제조Preparation of 100% (v / v) 2-propanol Extract-Activated Carbon Treatment-

100 g of fine leaf leaves were quenched with 800 ml of 100% (v / v) 2-propanol at room temperature for 20 hours, and circulated by filtration using a column filled with 5 g of activated carbon, and then again added to the residue 100% (v / v) 2 Add 800 ml of propanol, cool for 4 hours, and filter the column filled with 5 g of activated carbon. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<비교예 1> Comparative Example 1 원생약 대비 활성탄 100%(w/w) 처리 95%(v/v) 에탄올 추출물의 제조Preparation of 95% (v / v) Ethanol Extracts Treated with 100% (w / w) Activated Carbon

100 g of fine leaf leaves were immersed in 800 ml of 95% (v / v) ethanol at room temperature for 20 hours, and circulated by using a column filled with 100 g of activated carbon, and then the residue was again 800 ml of 95% (v / v) ethanol. After 4 hours of cooling and circulating filtration using a column filled with 100g of activated carbon. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<비교예 2> Comparative Example 2 원생약 대비 활성탄 50%(w/w) 처리 95%(v/v) 에탄올 추출물의 제조Preparation of 95% (v / v) Ethanol Extracts Treated with 50% (w / w) of Activated Carbon

100 g of fine leaf leaves were cooled to 800 ml of 95% (v / v) ethanol at room temperature for 20 hours, and circulated by filtration using a column filled with 50 g of activated carbon, and then the residue was again 800 ml of 95% (v / v) ethanol. After 4 hours of cooling and 50g of activated carbon filled column was filtered through circulation. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<비교예 3> Comparative Example 3 원생약 대비 활성탄 30%(w/w) 처리 95%(v/v) 에탄올 추출물의 제조Preparation of 95% (v / v) Ethanol Extracts Treated with 30% (w / w) of Activated Carbon

100 g of fine leaf leaves were immersed in 800 ml of 95% (v / v) ethanol at room temperature for 20 hours, and circulated through a column filled with 30 g of activated carbon, and the residue was again filtered into 800 ml of 95% (v / v) ethanol. After 4 hours of cooling, the column was filled with 30 g of activated carbon and filtered through a column. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<비교예 4> <Comparative Example 4> 70%(v/v) 에탄올 추출물-활성탄 미처리-의 제조Preparation of 70% (v / v) Ethanol Extract-Untreated Activated Carbon

100 g of fine leaf leaves were filtered by cooling with 800 ml of 70% (v / v) ethanol at room temperature for 20 hours, and then filtered into 800 ml of 70% (v / v) ethanol. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<비교예 5> Comparative Example 5 80%(v/v) 에탄올 추출물-활성탄 미처리-의 제조Preparation of 80% (v / v) Ethanol Extract-No Activated Carbon-

100 g of fine leaf leaves were cooled by cooling for 80 hours at room temperature with 80 ml of 80% (v / v) ethanol, and then filtered into 800 ml of 80% (v / v) ethanol. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<비교예 6> Comparative Example 6 85%(v/v) 에탄올 추출물-활성탄 미처리-의 제조Preparation of 85% (v / v) Ethanol Extract-Non-Activated Carbon-

100 g of fine leaf leaves were cooled by cooling for 20 hours at room temperature with 800 ml of 85% (v / v) ethanol, and then added to the residue by about 800 ml of 85% (v / v) ethanol for 4 hours and filtered. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<비교예 7> Comparative Example 7 90%(v/v) 에탄올 추출물-활성탄 미처리-의 제조Preparation of 90% (v / v) Ethanol Extract-Untreated Activated Carbon

100 g of the fine leaf leaves were filtered by cooling with 800 ml of 90% (v / v) ethanol at room temperature for 20 hours, and then filtered into 800 ml of 90% (v / v) ethanol. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<비교예 8> <Comparative Example 8> 95%(v/v) 에탄올 추출물-활성탄 미처리-의 제조Preparation of 95% (v / v) Ethanol Extract-No Activated Carbon

100 g of fine leaf leaves were filtered by cooling with 800 ml of 95% (v / v) ethanol at room temperature for 20 hours, and then filtered into 800 ml of 95% (v / v) ethanol. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<비교예 9> Comparative Example 9 100%(v/v) 에탄올 추출물-활성탄 미처리-의 제조Preparation of 100% (v / v) Ethanol Extract-Untreated Activated Carbon

100 g of fine leaf leaves were filtered by cooling with 800 ml of 100% (v / v) ethanol at room temperature for 20 hours, and then filtered again by adding 800 ml of 100% (v / v) ethanol to the residue for 4 hours. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<비교예 10> Comparative Example 10 100%(v/v) 1-프로판올 추출물-활성탄 미처리-의 제조Preparation of 100% (v / v) 1-propanol extract-no activated carbon

100 g of fine leaf leaves were cooled to 100 ml of 100% (v / v) 1-propanol at room temperature for 20 hours, and filtered. The residue was further cooled to 100 ml (v / v) 1-propanol for 4 hours. Filtered. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<비교예 11> Comparative Example 11 100%(v/v) 2-프로판올 추출물-활성탄 미처리-의 제조Preparation of 100% (v / v) 2-propanol extract-no activated carbon

100 g of fine leaf leaves were cooled with 100 ml of 100% (v / v) 2-propanol at room temperature for 20 hours and filtered. Then, the residue was added to 800 ml of 100% (v / v) 2-propanol and cooled for 4 hours. Filtered. The combined filtrates were concentrated under reduced pressure to obtain about 5 g of soft extract.

<실험예 1> Experimental Example 1 활성탄 처리 양에 따른 애엽 추출물 중 유해물질의 제거 효과와 유효성분 함량 변화 평가Evaluation of Removal Effect of Hazardous Substances and Changes in Active Ingredients of Leafy Leaf Extracts According to Activated Carbon

Comparative evaluation of 95% (v / v) ethanol leaf extracts (Examples 1 to 5 and Comparative Examples 1 to 3) treated with activated carbon to determine the changes in the content of benzopyrene and the active ingredient in the leaf extract according to the amount of activated carbon It was. 95% (v / v) ethanol leaf extract (Comparative Example 8) not treated with activated carbon was used as a control. Each soft extract was evaluated using high performance liquid chromatography (HPLC) and the results are shown in Table 1 and FIG. 1.

Table 1

As a result of the experiment, as shown in Table 1, the larvae extract (Comparative Examples 1 to 3) using activated carbon 30 ~ 100% (w / w) compared to the original drug can remove all benzopyrene, but it is an active ingredient Tilin and postiosidine also showed large content changes.

By the way, Examples 1 to 5 according to the present invention can be seen that there is no significant change in the content of eupatillin, and postiosidine while showing a benzopyrene reduction effect. In particular, as can be seen in Example 5, it can be seen that even if the amount of activated carbon compared to the original drug using 0.1% (w / w) benzopyrene is reduced by about 80% (w / w).

<실험예 2> Experimental Example 2 활성탄 처리 양에 따른 애엽 추출물의 위병변 억제 평가Evaluation of Gastric Lesion Inhibition of Leafy Leaf Extracts According to the Amount of Activated Carbon

95% (v / v) ethanol leaf extract in Experimental Example 1 showed a change in the content of harmful benzopyrene, the active ingredient eupatin, and postiosidine depending on the amount of activated carbon. Here, the protective effect of gastric mucosa was evaluated in a conventional hydrochloric acid-ethanol induced gastric injury rat model using the leaf extracts prepared in Examples 1 to 5, Comparative Examples 2, 3 and Comparative Example 8 (Comparative Example 1). Was removed from the experiment because all of the active ingredients were removed). The results are shown in Table 2 and FIG.

TABLE 2

As can be seen in Table 2, Comparative Examples 2 and 3 can remove all of the amount of benzopyrene, but the active ingredients, eupatilin, and postiosidine also showed a large content change, so that the effect of inhibiting gastric bleeding lesions was negligible

However, in Examples 1 to 5 using activated carbon at 20% (w / w) or less, it was found that the effect of inhibiting gastric bleeding lesions was not different from that of untreated activated carbon.

Therefore, the present invention, when using 0.1% (w / w) to 20% (w / w) of activated carbon compared to the original drug, while reducing the benzopyrene, a harmful substance compared to the activated carbon untreated group, the effect of inhibiting gastric bleeding lesions It can be seen that.

<실험예 3> Experimental Example 3 활성탄 적용 방법에 따른 애엽 추출물 중 유해물질의 제거 평가Evaluation of Removal of Hazardous Substances in Leafy Leaf Extracts According to Application Method of Activated Carbon

In the case of using 0.1% (w / w) to 20% (w / w) of activated carbon relative to the weight of the leaf of the crude leaf, which is a crude drug from Experimental Examples 1 and 2, the amount of benzopyrene, which is a toxic substance compared to the activated carbon untreated group, was reduced. It was found that the leaf extract showed the inhibitory effect on gastric bleeding lesions.

Here, the changes of the benzopyrene content and the active ingredient content according to the application method during the extraction process of the activated carbon of the activated carbon were investigated. 5% (w / w) of activated charcoal was used to compare circulating filtration during extraction (Example 3), post extraction filtration (Example 6), and post extraction redissolution filtration (Example 8). Each soft extract was evaluated using high performance liquid chromatography (HPLC) and the results are shown in Table 3.

TABLE 3

Experimental results As can be seen from Table 3, 5% (w / w) of activated carbon was used for circulating filtration during extraction (Example 3), filtration after extraction (Example 6), and after re-dissolution filtration after Example 8) In all the applied methods, the benzopyrene reduction rate and the eutectiline and postiosidine content reductions of the young leaf were similar in all solvents.

Therefore, it was found that the activated carbon filtration process in the present invention does not have a difference between application time points.

<실험예 4> Experimental Example 4 활성탄 필터를 사용한 용매별 애엽추출물 중 유해물질의 제거 평가Evaluation of Removal of Hazardous Substances in Leaf Extracts by Solvent Using Activated Carbon Filter

Comparative evaluation was performed for each solvent. Examples 3 and 8 to 14 were compared among the examples extracted using the activated carbon filtration process. Comparative Example 8, which was not treated with activated carbon, was used as a control. Each soft extract was evaluated using high performance liquid chromatography (HPLC) and the results are shown in Table 4.

Table 4

From the table 4, 70 ~ 100% (v / v) ethanol leaf extract, 100% (v / v) isopropanol leaf extract both benzopyrene reduction and eutectilin as a active ingredient of the leaf lobe, declining osmodine content of the present invention It can be seen that it is included in the category, there is no difference in the ability to reduce by extraction solvent when the activated carbon filtration process is applied, it is expected that it can exhibit the effect of inhibiting gastric hemorrhagic lesions of the same degree compared to the activated carbon untreated group.

Claims

A method for producing a leafy leaf extract for treating gastrointestinal diseases, in which leaflets are extracted with ethanol or isopropanol and reduced harmful substances by using 0.1-20% (w / w) activated carbon relative to the weight of leafy leaves.
[Claim 2] The method of claim 1, wherein the harmful substance is benzopyrene.
The method of claim 1, wherein the ethanol is 70 ~ 100% (v / v) method of producing a leaf extract for treating gastrointestinal disease, characterized in that ethanol.
The method of claim 1, wherein the isopropanol is 100% (v / v) 1-propanol or 100% (v / v) 2-propanol.
The method according to any one of claims 1 to 4, wherein the activated carbon is 0.1-5% (w / w) activated carbon based on the weight of the leaf of the leaf.
A method of reducing the benzopyrene content by using 0.1-20% (w / w) activated carbon relative to the weight of the leaf of the leaf, which is extracted from the leaf of the leaf with ethanol or propanol.
7. The method according to claim 6, wherein the step of reducing benzopyrene in the leaf extract is treated during extraction of the leaf of the leaf.
The method for reducing benzopyrene content according to claim 6, wherein the step of reducing benzopyrene in the leaf extract is treated before the concentration after extracting the leaf leaves.
The method for reducing benzopyrene content according to claim 6, wherein the step of reducing benzopyrene in the leaf extract is concentrated and re-dissolved.