KR102414382B1 - A method for preparing artemisia extract with a reduced content of benzopyrene - Google Patents

Abstract

Disclosed is a method for reducing benzopyrene in an extract of aerobium using a mixture of activated carbon and zeolite. According to the present invention, by mixing and processing zeolite and activated carbon, the content of the index component and the therapeutic effect of the aeyeop extract for the treatment of gastrointestinal diseases are maintained, and the benzopyrene that may be derived from the probiotic or generated during the manufacturing process is effectively reduced by reducing the aeyeop extract. safety can be ensured.

Description

A method for producing a leaf extract with reduced benzopyrene {A METHOD FOR PREPARING ARTEMISIA EXTRACT WITH A REDUCED CONTENT OF BENZOPYRENE}

The present invention relates to a method for producing an aeyeop extract used as an active ingredient for a treatment for gastrointestinal diseases. More specifically, it relates to a method for reducing benzopyrene, a harmful substance present in the extract, and relates to a method for producing an extract of an ae leaf in which benzopyrene, a harmful substance, is reduced by using activated carbon and zeolite.

Interest and concerns about harmful substances in herbal medicines are increasing. In particular, as heavy metals and residual pesticides in herbal medicines are detected, questions are raised about the safety of ingesting herbal medicines. In particular, as benzopyrene is detected in products such as ginseng, methods for removing harmful substances throughout herbal medicines are being studied, and regulations are expected to be gradually strengthened in drug approval institutions.

Benzopyrene is mainly produced when carbohydrates, fats, and proteins are incompletely burned during high-temperature treatment. It can also be found in unprocessed foods such as fruits and vegetables. It is known that the amount of benzopyrene generated during processing increases as cooking time close to fire or processing time increases. Accordingly, the drug approval institution has enacted a notice on benzopyrene acceptance standards and test methods for herbal medicines as a management plan for benzopyrene, a new hazardous substance, and is establishing management measures such as monitoring benzopyrene in distributed herbal or herbal medicines. Accordingly, there is a demand for reducing benzopyrene present in herbal medicines.

For example, Korean Patent Application Laid-Open No. 10-2015-0085330 discloses a method for producing an aeyeop extract for the treatment of gastrointestinal diseases with a reduced content of harmful substances. Disclosed is a method for reducing benzopyrene by using 0.1 to 20% (w/w) activated carbon relative to the weight of the leaf. In the above patent document, the technical feature is that benzopyrene is adsorbed by using activated carbon in the process of manufacturing the aeyeop extract. It is stated that it can be selectively adsorbed.

However, according to the research of the present inventor, the method described in the above patent document alone has a poor adsorption rate of benzopyrene present in the extract of a leaf extract. came to

Republic of Korea Patent Publication No. 10-2015-0085330

An object of the present invention is to provide a method capable of significantly reducing only benzopyrene without causing a change in the content of an indicator component by mixing and treating activated carbon and zeolite during the manufacturing process of the aeyeop extract.

As a technical means to solve the above problem, in the present invention, a method for reducing benzopyrene in ayeop extract is disclosed, characterized in that a mixture of zeolite and activated carbon is added and reacted by adding 0.04 to 40% (w/w) based on the weight of the aeyeop extract. . In addition, there is disclosed a method for producing ayeop extract comprising using the benzopyrene reduction method.

According to the present invention, it is possible to selectively adsorb only benzopyrene, which is a harmful substance, without causing a change in the content of eupatylin or zeosidine, which are index components, in the extract of the leaf extract. The selective adsorption rate is significantly increased.

1 and 2 are diagrams confirming the component profile of the extract of the treated and not treated with a mixture of zeolite and activated carbon.

Short-term exposure to benzopyrene may cause teratogenic and immunosuppressive reactions during pregnancy, and long-term exposure may cause local carcinogenicity in animals by skin contact, inhalation, intratracheal and bronchial administration, subcutaneous or intramuscular injection, and other uses. is known Oral administration to male F344 rats showed a decrease in leukocytes and an increase in mean cell-hemoglobin concentration in an acute toxicity study. In addition, the liver weight to body weight ratio increased by up to 30%. The chronic toxicity test showed a decrease in body weight and an increase in the liver-to-weight ratio, and in the high-dose group, abnormalities in the kidneys and BUN among chemical factors increased. (Knuckles et al., 2001) In addition, the US EPA classified benzopyrene as B2 because it is thought to be carcinogenic to humans, and it has been proven carcinogenic in a number of animal experiments.

Mugwort (Artemisia mongolica, A. asiatica, A. princeps var. orientalis, A. argyi, A. montana, etc.) is a perennial herb belonging to the Asteraceae family and has been used as a tonic blood, gynecological disease, and diarrhea treatment since ancient times. As such, there are isocoumarin, coumarin, diterpene lactone, and the like, and flavonoid components such as eupatylline and zeosidine were also confirmed. In addition to the Korean Pharmacopoeia, the herbal medicine (herbal medicine) standard is referred to as aeyeop (艾葉, Artemisiae argyi Herb) as the leaves and young stems of the mugwort. Among them, eupatilin, a flavonoid component, is known for its anti-allergic effect (Japanese Patent Application Laid-Open No. 59-155314), and its efficacy as a treatment for gastrointestinal diseases has also been revealed in Korea (Korean Patent Registration 127777). Another flavonoid component, jaseocidin, has also been registered as a patent for use as a treatment for the same gastrointestinal disease. (Korea Patent Registration 181751) In the present invention, in the preparation of the ayeop extract, a mixture of zeolite and activated carbon is added to the aeyeop extract obtained by extracting the ayeop extract with ethanol or isopropanol, and the mixture is stirred and filtered, or the mixture is filled in a column By using it, it is characterized in that it selectively removes benzopyrene present in the leaf extract.

According to the present invention, by mixing and processing zeolite and activated carbon, the content of the index component and the therapeutic effect of the aeyeop extract for the treatment of gastrointestinal diseases are maintained, and the benzopyrene that may be derived from the probiotic or generated during the manufacturing process is effectively reduced by reducing the aeyeop extract. safety can be ensured.

At this time, the mixing ratio of activated carbon and zeolite may be 1:5 to 5:1, and it is preferable to use it by mixing 1:1.

Activated carbon is a material whose main component is carbon and is porous, so it has a large surface area, has strong adsorption, and a chemical reaction occurs quickly. In the present invention, while selectively adsorbing only benzopyrene, indicator components ( ^eupatylin , zeosidine) should not be adsorbed. do.

Zeolite, as a crystalline aluminum silicate mineral, has the property of selectively and strongly adsorbing unsaturated hydrocarbons or polar substances by the action of cations in its crystal structure, and also has a pore diameter of a certain size. It can be selectively passed through and adsorbed. The zeolite used in the present invention preferably has a specific surface area of 200 m ² /g or more, a micropore area of 120 m ² /g or more, and a Si/Al molecular ratio of 2.5 or more.

In the present invention, a mixture of activated carbon and zeolite is used. That is, according to the present inventor's study, activated carbon has an excellent effect of reducing benzopyrene, but when it is used in excess of 20%, the content change of the indicator component is large. There is this. On the other hand, it was found that zeolite had a lower benzopyrene removal effect than activated carbon, but did not significantly affect the index component content. In the present invention, by mixing and treating activated carbon and zeolite, a method for remarkably reducing benzopyrene without a large content change in the indicator component is proposed. Hereinafter, the present invention will be described by way of Examples.

Comparative Example 1) Preparation of extracts not treated with zeolite or activated carbon

100 g of chopped ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. After filtering the primary concentrate, it was concentrated under reduced pressure to obtain about 5 g of soft crude extract.

Comparative Example 2) Preparation of extract treated with zeolite 0.02%

100 g of minced ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 0.02 g of zeolite was added to the primary concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. As the zeolite, a zeolite having a specific surface area of 403 m ² /g, a micropore area of 241 m ² /g, and a Si/Al molecular ratio of 25 was used.

Comparative Example 3) Preparation of extract treated with zeolite 0.2%

100 g of minced ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 0.2 g of zeolite was added to the primary concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. As the zeolite, a zeolite having a specific surface area of 403 m ² /g, a micropore area of 241 m ² /g, and a Si/Al molecular ratio of 25 was used.

Comparative Example 4) Preparation of extract treated with zeolite 2%

100 g of chopped ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 2 g of zeolite was added to the primary concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. As the zeolite, a zeolite having a specific surface area of 403 m ² /g, a micropore area of 241 m ² /g, and a Si/Al molecular ratio of 25 was used.

Comparative Example 5) Preparation of extract treated with 20% zeolite

100 g of chopped ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 20 g of zeolite was added to the first concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. As the zeolite, a zeolite having a specific surface area of 403 m ² /g, a micropore area of 241 m ² /g, and a Si/Al molecular ratio of 25 was used.

Comparative Example 6) Preparation of an extract treated with 40% zeolite

100 g of minced ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 40 g of zeolite was added to the first concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. As the zeolite, a zeolite having a specific surface area of 403 m ² /g, a micropore area of 241 m ² /g, and a Si/Al molecular ratio of 25 was used.

Comparative Example 7) Preparation of extract treated with activated carbon 0.02%

100 g of minced ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 0.02 g of activated carbon was added to the primary concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. Activated carbon having a specific surface area of 1,200 m ² /g and a particle size of 74um or less (200mesh or more) was used.

Comparative Example 8) Preparation of extract treated with activated carbon 0.2%

100 g of minced ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 0.2 g of activated carbon was added to the primary concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. Activated carbon having a specific surface area of 1,200 m ² /g and a particle size of 74um or less (200mesh or more) was used.

Comparative Example 9) Preparation of extract treated with activated carbon 2%

100 g of chopped ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 2 g of activated carbon was added to the primary concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. Activated carbon having a specific surface area of 1,200 m ² /g and a particle size of 74um or less (200mesh or more) was used.

Comparative Example 10) Preparation of extract treated with 10% activated carbon

100 g of chopped ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 10 g of activated carbon is added to the primary concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft shell extract. Activated carbon having a specific surface area of 1,200 m ² /g and a particle size of 74um or less (200mesh or more) was used.

Comparative Example 11) Preparation of an extract treated with 40% activated carbon

100 g of minced ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 40 g of activated carbon was added to the first concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. Activated carbon having a specific surface area of 1,200 m ² /g and a particle size of 74um or less (200mesh or more) was used.

Comparative Example 12) Preparation of extract treated with 80% of a mixture of zeolite and activated carbon

100 g of minced ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 80 g of a mixture of zeolite and activated carbon (zeolite: activated carbon = 1:1) was added to the primary concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. As the zeolite, a zeolite having a specific surface area of 403 m ² /g, a micropore area of 241 m ² /g, and a Si/Al molecular ratio of 25 was used. Activated carbon having a specific surface area of 1,200 m ² /g and a particle size of 74um or less (200mesh or more) was used.

Comparative Example 13) Preparation of extracts washed with protozoa

100g of chopped ae leaves are soaked in lukewarm water, washed, and blow-dried at 40°C. The dried ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. After filtering the primary concentrate, it was concentrated under reduced pressure to obtain about 5 g of soft crude extract.

Comparative Example 14) Preparation of extract treated with 0.2% activated carbon after washing the protozoan

100g of chopped ae leaves are soaked in lukewarm water, washed, and blow-dried at 40°C. The dried ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 0.2 g of activated carbon was added to the primary concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. Activated carbon having a specific surface area of 1,200 m ² /g and a particle size of 74um or less (200mesh or more) was used.

Comparative Example 15) Preparation of extract treated with zeolite 0.2%

100 g of minced ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary and secondary leachates were combined and concentrated under reduced pressure to obtain 240 ml of a primary concentrate. To the primary concentrate, 0.2 g of zeolite was added, stirred for 4 hours, filtered, and concentrated under reduced pressure to obtain about 5 g of soft crude extract. As the zeolite, a zeolite having a specific surface area of 198 m ² /g, a micropore area of 116 m ² /g, and a Si/Al molecular ratio of 2 was used.

Comparative Example 16) Preparation of extract treated with activated carbon 0.2%

100 g of chopped ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 0.2 g of activated carbon was added to the primary concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. Activated carbon having a specific surface area of 580 m ² /g and a particle size of 177 μm or less (80 mesh or more) was used as the activated carbon.

Comparative Example 17) Preparation of extract treated with zeolite and activated carbon mixture 0.4%

100 g of minced ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 0.4 g of a mixture of zeolite and activated carbon (zeolite: activated carbon = 1:1) was added to the primary concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. As the zeolite, a zeolite having a specific surface area of 198 m ² /g, a micropore area of 116 m ² /g, and a Si/Al molecular ratio of 2 was used. Activated carbon having a specific surface area of 580 m ² /g and a particle size of 177 μm or less (80 mesh or more) was used as the activated carbon.

Example 1) Preparation of extract treated with zeolite and activated carbon mixture 0.04%

100 g of minced ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 0.04 g of a mixture of zeolite and activated carbon (zeolite: activated carbon = 1:1) was added to the primary concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. As the zeolite, a zeolite having a specific surface area of 403 m ² /g, a micropore area of 241 m ² /g, and a Si/Al molecular ratio of 25 was used. Activated carbon having a specific surface area of 1,200 m ² /g and a particle diameter of 74 μm or less (200 mesh or more) was used as the activated carbon.

Example 2) Preparation of extract treated with zeolite and activated carbon mixture 0.4%

100 g of chopped ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 0.4 g of a mixture of zeolite and activated carbon (zeolite: activated carbon = 1:1) was added to the primary concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. As the zeolite, a zeolite having a specific surface area of 403 m ² /g, a micropore area of 241 m ² /g, and a Si/Al molecular ratio of 25 was used. Activated carbon having a specific surface area of 1,200 m ² /g and a particle size of 74um or less (200mesh or more) was used.

Example 3) Preparation of extract treated with zeolite and activated carbon mixture 4%

100 g of minced ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 4 g of a mixture of zeolite and activated carbon (zeolite: activated carbon = 1:1) was added to the primary concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. As the zeolite, a zeolite having a specific surface area of 403 m ² /g, a micropore area of 241 m ² /g, and a Si/Al molecular ratio of 25 was used. Activated carbon having a specific surface area of 1,200 m ² /g and a particle size of 74um or less (200mesh or more) was used.

Example 4) Preparation of an extract treated with 40% of a mixture of zeolite and activated carbon

100 g of chopped ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 40 g of a mixture of zeolite and activated carbon (zeolite: activated carbon = 1:1) was added to the primary concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. As the zeolite, a zeolite having a specific surface area of 403 m ² /g, a micropore area of 241 m ² /g, and a Si/Al molecular ratio of 25 was used. Activated carbon with a specific surface area of 1,200 m ² /g and a particle size of 74um or less (200mesh or more) was used.

Example 5) Preparation of extract using a column filled with a mixture of zeolite and activated carbon 0.4%

100 g of minced ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. The primary concentrate was mixed with 0.2 g each of zeolite and activated carbon, filtered through continuous circulation in a column packed, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. As the zeolite, a zeolite having a specific surface area of 403 m ² /g, a micropore area of 241 m ² /g, and a Si/Al molecular ratio of 25 was used. Activated carbon having a specific surface area of 1,200 m ² /g and a particle size of 74um or less (200mesh or more) was used.

Example 6) Preparation of extract treated with 0.4% of a mixture of zeolite and activated carbon after washing the protozoan

100g of chopped ae leaves are soaked in lukewarm water, washed, and blow-dried at 40°C. The dried ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 0.4 g of a mixture of zeolite and activated carbon (zeolite: activated carbon = 1:1) was added to the primary concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. As the zeolite, a zeolite having a specific surface area of 403 m ² /g, a micropore area of 241 m ² /g, and a Si/Al molecular ratio of 25 was used. Activated carbon having a specific surface area of 1,200 m ² /g and a particle size of 74um or less (200mesh or more) was used.

Example 7) Preparation of extract treated with zeolite and activated carbon mixture 0.4%

100 g of minced ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 0.4 g of a mixture of zeolite and activated carbon (zeolite: activated carbon = 1: 5) was added to the primary concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. As the zeolite, a zeolite having a specific surface area of 403 m ² /g, a micropore area of 241 m ² /g, and a Si/Al molecular ratio of 25 was used. Activated carbon having a specific surface area of 1,200 m ² /g and a particle size of 74um or less (200mesh or more) was used.

Example 8) Preparation of extract treated with zeolite and activated carbon mixture 0.4%

100 g of minced ae leaves were cooled with 1,700 ml of 100% isopropanol at room temperature for 24 hours and filtered (first leachate). The primary leachate and the secondary leachate were combined and concentrated under reduced pressure to obtain 240ml of the primary concentrate. 0.4 g of a mixture of zeolite and activated carbon (zeolite: activated carbon = 5: 1) was added to the primary concentrate, stirred for 4 hours, filtered, and then concentrated under reduced pressure to obtain about 5 g of soft crude extract. As the zeolite, a zeolite having a specific surface area of 403 m ² /g, a micropore area of 241 m ² /g, and a Si/Al molecular ratio of 25 was used. Activated carbon having a specific surface area of 1,200 m ² /g and a particle diameter of 74 μm or less (200 mesh or more) was used as the activated carbon.

Experimental Example 1) benzopyrene reduction and index component content evaluation according to the dose of zeolite or activated carbon

In order to confirm the effect of reducing benzopyrene according to the amount of zeolite or activated carbon and the change in the content of the indicator component, isopropanol aerobium extract treated with different doses of zeolite or activated carbon was comparatively evaluated. Comparative Example 1 not treated with activated carbon was used as a control. Each extract was evaluated using high performance liquid chromatography.

treatment group absorbent Adsorbent input amount compared to crude drug (%) compared to control Benzopyrene (%) Eupatylline (%) Zeiocidin (%) Comparative Example 1 unprocessed unprocessed 100 100 100 Comparative Example 2 zeolite 0.02 72 100 99 Comparative Example 3 0.2 59 98 100 Comparative Example 4 2 48 99 99 Comparative Example 5 20 47 98 99 Comparative Example 6 40 47 97 98 Comparative Example 7 activated carbon 0.02 37 98 98 Comparative Example 8 0.2 26 96 95 Comparative Example 9 2 14 90 91 Comparative Example 10 10 10 80 80 Comparative Example 11 40 0 52 51

When zeolite was treated with 0.02 to 40% (w/w), the benzopyrene residual rate did not decrease any more in the group treated with 2% (w/w) or more, and the contents of eupatylin and zeosidine, which are index components, showed no significant change. there was no

When the activated carbon was treated with 0.02-40% (w/w), the benzopyrene residual rate decreased as the dose increased, and in the case of eupatylin and zeosidine, which are indicator ingredients, a large content change was observed in the treatment group exceeding 20% (w/w). showed In the case of the activated carbon-treated group, the benzopyrene reduction effect was higher than that of the zeolite-treated group.

Experimental Example 2) benzopyrene reduction and index component content evaluation by mixing zeolite and activated carbon

A comparative evaluation was performed according to the mixing of zeolite and activated carbon in order to confirm the effect of reducing benzopyrene and the change in the content of index components according to the mixing of zeolite and activated carbon. Comparative Example 1 not treated with zeolite or activated carbon was used as a control. Each extract was evaluated using high performance liquid chromatography.

treatment group absorbent Adsorbent input amount compared to crude drug (%) compared to control Benzopyrene (%) Eupatylline (%) Zeiocidin (%) Comparative Example 1 unprocessed unprocessed 100 100 100 Comparative Example 7 activated carbon 0.02 37 98 98 Comparative Example 8 0.2 26 96 95 Comparative Example 9 2 14 90 91 Comparative Example 10 10 10 80 80 Comparative Example 11 40 0 52 51 Example 1 Zeolite and activated carbon mixture 0.04 4 99 98 Example 2 0.4 0 96 96 Example 3 4 0 91 91 Example 4 40 0 80 81 Comparative Example 12 80 0 51 50

In the mixture of zeolite and activated carbon, the effect of reducing benzopyrene was significantly increased compared to the single treatment of activated carbon, and there was no significant change in the contents of eupatylin and zeosidine, which are index components, in all treatment groups except 80% (w/w). In particular, in the 0.4% (w/w) treatment group of a mixture of zeolite and activated carbon, benzopyrene was significantly removed without significant change in the content of eupatylline and zeosidine, which are index components.

Experimental Example 3) Reduction of benzopyrene by using a column and evaluation of index component content

Circulation filtration was performed on a column filled with a mixture of zeolite and activated carbon to confirm the effect of reducing benzopyrene and change in the content of indicator components. In addition, the benzopyrene reduction effect and the change in the content of the indicator components were confirmed when the activated carbon or a mixture of zeolite and activated carbon was treated after washing and drying of the probiotic. Comparative Example 1 not treated with zeolite or activated carbon was used as a control. Each extract was evaluated using high performance liquid chromatography.

treatment group absorbent Adsorbent input amount compared to crude drug (%) compared to control Benzopyrene (%) Eupatylline (%) Zeiocidin (%) Comparative Example 1 unprocessed unprocessed 100 100 100 Comparative Example 8 activated carbon 0.2 26 96 95 Example 2 Zeolite and activated carbon mixture 0.4 0 96 96 Example 5 Zeolite, activated carbon column 0.4 0 95 96 Comparative Example 13 Probiotic cleaning unprocessed 76 100 100 Comparative Example 14 Activated carbon after washing the raw material 0.2 20 95 95 Example 6 A mixture of zeolite and activated carbon after washing the protozoan 0.4 0 96 95

When circulation filtration was performed on a column filled with a mixture of zeolite and activated carbon, it exhibited an effect of reducing benzopyrene at the same level as when treating a mixture of zeolite and activated carbon, and there was no significant change in the contents of eupatylin and zeosidine, which are index components. .

When the crude drug was washed and dried, the residual rate of benzopyrene was reduced to 76% compared to untreated, but when the mixture of zeolite and activated carbon was treated, the same benzopyrene reduction effect was shown regardless of washing or unwashing.

Experimental Example 4) benzopyrene reduction and index component content evaluation according to the physical properties of zeolite and activated carbon

The effect of reducing benzopyrene and changes in the content of index components according to the physical properties of zeolite and activated carbon were confirmed. Comparative Example 1, which was not treated with zeolite or activated carbon, was used as a control group. Each extract was evaluated using high performance liquid chromatography.

treatment group absorbent Adsorbent input amount compared to crude drug (%) compared to control Benzopyrene (%) Eupatylline (%) Zeiocidin (%) Comparative Example 1 unprocessed unprocessed 100 100 100 Comparative Example 3 zeolite 0.2 59 98 100 Comparative Example 15 98 99 100 Comparative Example 8 activated carbon 0.2 26 96 95 Comparative Example 16 88 98 99 Example 2 Zeolite and activated carbon mixture 0.4 0 96 96 Comparative Example 17 89 99 98

The comparative example confirmed that the benzopyrene reduction effect was significantly lower than that of the Example.

Experimental Example 5) benzopyrene reduction and index component content evaluation according to the mixing ratio of zeolite and activated carbon

A comparative evaluation was performed according to the mixing ratio of zeolite and activated carbon to confirm the effect of reducing benzopyrene and the change in the content of index components according to the mixing ratio of zeolite and activated carbon. Comparative Example 1 not treated with zeolite or activated carbon was used as a control. Each extract was evaluated using high performance liquid chromatography.

treatment group absorbent Adsorbent input amount compared to raw drug (%) mixing ratio compared to control Benzopyrene (%) Eupatylline (%) Zeiocidin (%) Comparative Example 1 unprocessed unprocessed - 100 100 100 Comparative Example 8 activated carbon 0.2 - 26 96 95 Example 7 Zeolite and activated carbon mixture 0.4 1: 5 0 96 95 Example 8 5 : 1 0 97 97 Example 2 1:1 0 96 96

At a mixing ratio of zeolite and activated carbon of 1: 5 to 5: 1, the effect of reducing benzopyrene was significantly increased compared to the single treatment of activated carbon.

Experimental Example 6) Gastric lesion inhibitory effect evaluation

Gastrointestinal disease treatment effect was evaluated for the aeyeop extract according to each example. Comparative Example 1 in which activated carbon and zeolite were not treated was used as a control. As an animal model for evaluating the treatment effect of gastrointestinal diseases, a hydrochloric acid-ethanol-induced gastric injury rat model was used, and the treatment effect was evaluated by measuring the rate of gastric lesions.

treatment group absorbent Adsorbent input amount compared to raw drug (%) number of animals compared to the disease-inducing group
Stomach lesion area (%) disease induction group - - 10 100 Comparative Example 1 unprocessed unprocessed 10 14** Example 1 Zeolite and activated carbon mixture 0.04 10 15** Example 2 0.4 10 14** Example 3 4 10 13** Example 4 40 10 13** Comparative Example 12 80 10 48 Example 5 Zeolite, Activated Carbon Column 0.4 10 14**

** Statistical significance compared to disease induction group (P<0.01)

As can be seen from Table 6, Comparative Example 12 can remove all of the amount of benzopyrene, but the active ingredients eupatylline and zeosidine also showed a large content change, so the effect of inhibiting gastric bleeding lesions was insignificant. Examples 1 to 4 using a mixture of zeolite and activated carbon at 40% (w/w) or less and Example 5, in which a column filled with a mixture of zeolite and activated carbon was filtered through circulation, showed no difference in the effect of inhibiting gastric bleeding lesion compared to the activated carbon untreated group Could know. Therefore, the present invention uses 0.04% (w/w) to 40% (w/w) of a zeolite and activated carbon mixture compared to the protozoan, or in the case of circulation filtration in a column filled with a zeolite and activated carbon mixture, compared to the zeolite-activated carbon mixture untreated group It can be seen that while reducing benzopyrene, which is a harmful substance, the aeyeop extract exhibits an effect of inhibiting gastric bleeding lesions to the same extent.

Experimental Example 7) Evaluation of component profile equivalence of extracts

When the mixture of zeolite and activated carbon was treated, comparison between untreated extracts was evaluated to determine whether the component profile of the extract was changed. The results are shown in Table 7 below.

division RT Area correlation coefficient 0.9999 0.9820

The equicorrelation coefficient was 0.9 or more, and the component profiles of the extracts were the same when the mixture of zeolite and activated carbon was not treated and when treated.

According to the present invention, it is possible to prepare an ayeop extract with reduced benzopyrene.

Claims (7)

A method for reducing benzopyrene in Ayeop extract using a mixture of activated carbon and zeolite. The method according to claim 1, wherein a mixing ratio of activated carbon to zeolite in the mixture is 1:5-5:1. [3] The method according to claim 1 or 2, wherein the mixture is added to the aerobium extract and stirred. The method according to claim 1 or 2, wherein the mixture is used by filling a column. The method according to claim 1 or 2, wherein the mixture is used in an amount of 0.04 to 40% (w/w) based on the weight of the leaf. The method according to claim 5, wherein the mixture is used in an amount of 0.4 to 4% (w/w) based on the weight of the leaf. A method for preparing an ayeop extract by extracting the aeyeop with ethanol or isopropanol, comprising the method defined in claim 1 or 2, for producing an aeyeop extract.

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