KR102607410B1 - Antibacterial composition containing extracts of above-ground parts of glechoma grandis and skin external preparation containing the same - Google Patents
Antibacterial composition containing extracts of above-ground parts of glechoma grandis and skin external preparation containing the same Download PDFInfo
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- KR102607410B1 KR102607410B1 KR1020210088931A KR20210088931A KR102607410B1 KR 102607410 B1 KR102607410 B1 KR 102607410B1 KR 1020210088931 A KR1020210088931 A KR 1020210088931A KR 20210088931 A KR20210088931 A KR 20210088931A KR 102607410 B1 KR102607410 B1 KR 102607410B1
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- antibacterial
- extract
- centella asiatica
- test
- antibacterial composition
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- 235000012239 silicon dioxide Nutrition 0.000 description 1
- 230000037384 skin absorption Effects 0.000 description 1
- 231100000274 skin absorption Toxicity 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000005063 solubilization Methods 0.000 description 1
- 230000007928 solubilization Effects 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 235000010199 sorbic acid Nutrition 0.000 description 1
- 239000004334 sorbic acid Substances 0.000 description 1
- 229940075582 sorbic acid Drugs 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 239000010421 standard material Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 229940034610 toothpaste Drugs 0.000 description 1
- 239000000606 toothpaste Substances 0.000 description 1
- 238000011200 topical administration Methods 0.000 description 1
- 239000000196 tragacanth Substances 0.000 description 1
- 235000010487 tragacanth Nutrition 0.000 description 1
- 229940116362 tragacanth Drugs 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000002485 urinary effect Effects 0.000 description 1
- 229940120293 vaginal suppository Drugs 0.000 description 1
- 239000006216 vaginal suppository Substances 0.000 description 1
- 229940124549 vasodilator Drugs 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/005—Antimicrobial preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Dermatology (AREA)
- Botany (AREA)
- Epidemiology (AREA)
- Birds (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Engineering & Computer Science (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
본 발명은 긴병꽃풀 지상부 추출물을 포함하는 항균제 조성물 및 이를 포함하는 피부 외용제에 관한 것으로, 본 발명의 일 측면은, 긴병꽃풀(Glechoma grandis (A. Gray) Kuprian) 지상부 추출물을 유효성분으로 포함하는, 항균제 조성물을 제공한다.The present invention relates to an antibacterial composition containing an extract of the aerial part of the aerial part of the plant and an external skin preparation containing the same. One aspect of the present invention includes the extract of the aerial part of the aerial part of the plant , Glechoma grandis (A. Gray) Kuprian, as an active ingredient. Provides an antibacterial composition.
Description
본 발명은 긴병꽃풀 지상부 추출물을 포함하는 항균제 조성물 및 이를 포함하는 피부 외용제에 관한 것이다.The present invention relates to an antibacterial composition containing an extract of the aerial parts of Centella asiatica and an external skin preparation containing the same.
생활 속 항균 제품들은 대부분 화학물질(화학 항균제)들을 함유하고 있으며, 이로 인한 안전성 문제들이 지속적으로 발생하고 있다. 일례로, 우리나라에서도 얼마 전 '가습기 살균제'로 인해 심각한 사회적 문제가 발생한 바 있다. Most antibacterial products in everyday life contain chemical substances (chemical antibacterial agents), and safety problems continue to arise as a result. For example, in Korea, a serious social problem occurred recently due to 'humidifier disinfectant'.
그러나 현재 화학 항균제를 대체할 천연 물질 성분의 제품들은 생산되고 있지 않으며, 의약품 분야에서도 외용 항균제 대부분은 화학물질 기반의 항균 물질이 차지하고 있다. 따라서, 천연 물질 기반 항균제에 대한 개발이 신속하게 이루어질 필요가 있다.However, currently, products made of natural substances to replace chemical antibacterial agents are not being produced, and even in the pharmaceutical field, most of the external antibacterial agents are chemical-based antibacterial substances. Therefore, there is a need to rapidly develop antibacterial agents based on natural substances.
한편, 긴병꽃풀(Glechoma grandis (A. Gray) Kuprian)은 경기도와 대한민국 중남부 전역의 들판이나 논가에 자라는 여러해살이 초본으로, 한국에서는 금전초의 대용으로도 사용하였다. 한의학 문헌에서 언급되는 ‘금전초’는 앵초과의 과로황(Lysimachia christinae Hance)의 전초이며, 청열해독, 통림, 소종 등의 효능을 가지고 있어 간담질환과 비뇨기계의 결석을 치료하는데 쓰인다. Meanwhile, Glechoma grandis (A. Gray) Kuprian ) is a perennial herb that grows in fields and rice fields throughout Gyeonggi-do and south-central Korea, and is also used as a substitute for money plant in Korea. 'Geumjeoncho' mentioned in Oriental medicine literature is a precursor to Lysimachia christinae Hance of the primrose family, and has effects such as clearing heat, detoxification, pain relief, and somatocele, and is used to treat hepatobiliary diseases and stones in the urinary system.
그러나 아직까지 긴병꽃풀이 항균제 또는 항균용 피부 외용제로 사용된 사례는 보고된 바 없다.However, there have been no reported cases of Centella asiatica being used as an antibacterial agent or external antibacterial skin agent.
전술한 배경기술은 발명자가 본원의 개시 내용을 도출하는 과정에서 보유하거나 습득한 것으로서, 반드시 본 출원 전에 일반 공중에 공개된 공지기술이라고 할 수는 없다.The above-mentioned background technology is possessed or acquired by the inventor in the process of deriving the disclosure of the present application, and cannot necessarily be said to be known technology disclosed to the general public before the present application.
본 발명은 상술한 문제점을 해결하기 위한 것으로, 본 발명의 목적은, 긴병꽃풀 지상부 추출물을 함유하는 천연 물질 기반의 항균제 조성물 및 이를 포함하는 항균용 피부 외용제를 제공하는 것이다.The present invention is intended to solve the above-mentioned problems, and the purpose of the present invention is to provide an antibacterial composition based on natural substances containing an extract of the above-mentioned parts of Centella asiatica and an antibacterial external skin preparation containing the same.
본 발명의 다른 목적은, 항균제 조성물 제조를 위한 긴병꽃풀 지상부 추출물의 제조방법을 제공하는 것이다.Another object of the present invention is to provide a method for producing an extract of the above-ground parts of Centella asiatica for producing an antibacterial composition.
그러나, 본 발명이 해결하고자 하는 과제는 이상에서 언급한 것들로 제한되지 않으며, 언급되지 않은 또 다른 과제들은 아래의 기재로부터 해당 분야 통상의 기술자에게 명확하게 이해될 수 있을 것이다.However, the problems to be solved by the present invention are not limited to those mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.
본 발명의 일 측면은, 긴병꽃풀(Glechoma grandis (A. Gray) Kuprian) 지상부 추출물을 유효성분으로 포함하는, 항균제 조성물을 제공한다.One aspect of the present invention provides an antibacterial composition comprising an extract of the aerial parts of Glechoma grandis (A. Gray) Kuprian as an active ingredient.
일 실시형태에 따르면, 상기 긴병꽃풀 지상부 추출물의 농도는, 0.01 g/ml 내지 0.05 g/ml인 것일 수 있다.According to one embodiment, the concentration of the above-mentioned extract of the aerial parts of Centella asiatica may be 0.01 g/ml to 0.05 g/ml.
일 실시형태에 따르면, 상기 긴병꽃풀 지상부 추출물은, 폴리페놀 5 중량% 내지 10 중량%를 포함하는 것일 수 있다.According to one embodiment, the extract of the above-ground parts of Centella asiatica may contain 5% to 10% by weight of polyphenol.
일 실시형태에 따르면, 상기 긴병꽃풀 지상부 추출물은, 물, 탄소수 1 내지 4의 저급 알코올, 아세톤, 에틸아세테이트, 클로로포름, 1,3-부틸렌글리콜, n-헥산, 디에틸에테르, 부틸아세테이트 및 디클로로메탄으로 이루어진 군에서 선택되는 하나 이상을 추출 용매로 하여 추출된 것일 수 있다.According to one embodiment, the extract of the above-ground part of Centella asiatica is composed of water, lower alcohol having 1 to 4 carbon atoms, acetone, ethyl acetate, chloroform, 1,3-butylene glycol, n-hexane, diethyl ether, butyl acetate, and It may be extracted using at least one selected from the group consisting of dichloromethane as an extraction solvent.
일 실시형태에 따르면, 상기 저급 알코올은, 에탄올인 것일 수 있다.According to one embodiment, the lower alcohol may be ethanol.
일 실시형태에 따르면, 상기 추출은, 20 ℃ 내지 30 ℃의 온도에서 24 시간 내지 48 시간 동안 수행되는 것일 수 있다.According to one embodiment, the extraction may be performed at a temperature of 20°C to 30°C for 24 to 48 hours.
일 실시형태에 따르면, 폐렴간균(Klebsiella pneumoniae), 녹농균(Pseudomonas aeruginosa), 포도상구균(Staphylococcus aureus) 및 대장균(Escherichia coli) 중 적어도 하나 이상의 균주에 대하여 항균 활성을 갖는 것일 수 있다.According to one embodiment, it may have antibacterial activity against at least one strain of Klebsiella pneumoniae , Pseudomonas aeruginosa , Staphylococcus aureus , and Escherichia coli .
일 실시형태에 따르면, 상기 항균제 조성물은, 용액, 현탁액, 유탁액, 페이스트, 겔, 크림, 로션, 파우더, 비누, 계면활성제 함유 클렌징, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션 및 스프레이로 이루어진 군에서 선택되는 하나 이상의 제형을 포함하는 것일 수 있다.According to one embodiment, the antibacterial composition consists of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, and spray. It may include one or more formulations selected from the group.
본 발명의 다른 측면은, 상기 항균제 조성물을 포함하는, 항균용 피부 외용제를 제공한다.Another aspect of the present invention provides an antibacterial external skin preparation comprising the antibacterial composition.
본 발명에 따른 항균제 조성물은, 긴병꽃풀 지상부 추출물을 포함함으로써, 대장균, 황색포도상구균, 폐렴간균 및 녹농균에 대한 우수한 항균력을 갖는 효과가 있으며, 인체 안전성 및 피부 안전성이 확보될 수 있는 효과가 있다.The antibacterial composition according to the present invention, by containing the extract of the above-ground parts of Longhorn Calliflora, has the effect of having excellent antibacterial activity against Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, and Pseudomonas aeruginosa, and has the effect of ensuring human safety and skin safety. .
또한, 긴병꽃풀 지상부 추출물을 활용함으로써, 산림자원의 고부가가치화 및 다양한 제품 적용에 따른 신소득 창출에 기여할 수 있는 효과가 있다.In addition, by using extracts of the above-ground parts of Long-distance Lily, there is an effect of contributing to the creation of new income through high value-added forest resources and application of various products.
도 1은, 긴병꽃풀 에탄올 추출물(0.01 g/mL)의 대장균 및 황색포도상구균에 대한 항균시험 결과를 나타낸 사진이다.
도 2는, 긴병꽃풀 에탄올 추출물(0.02 g/mL)의 대장균 및 황색포도상구균에 대한 항균시험 결과를 나타낸 사진이다.
도 3은, 긴병꽃풀 에탄올 추출물(0.03 g/mL)의 대장균 및 황색포도상구균에 대한 항균시험 결과를 나타낸 사진이다.
도 4는, 긴병꽃풀 에탄올 추출물(0.05 g/mL)의 대장균 및 황색포도상구균에 대한 항균시험 결과를 나타낸 사진이다.
도 5는, 긴병꽃풀 에탄올 추출물(0.05 g/mL)의 녹농균 및 폐렴간균에 대한 항균시험 결과를 나타낸 사진이다.
도 6은, 긴병꽃풀 에탄올 추출물(0.03 g/mL)의 녹농균 및 폐렴간균에 대한 항균시험 결과를 나타낸 사진이다.
도 7은, 긴병꽃풀 에탄올 추출물(0.02 g/mL)의 녹농균 및 폐렴간균에 대한 항균시험 결과를 나타낸 사진이다.
도 8은, 긴병꽃풀 에탄올 추출물(0.01 g/mL)의 녹농균 및 폐렴간균에 대한 항균시험 결과를 나타낸 사진이다.Figure 1 is a photograph showing the results of an antibacterial test against Escherichia coli and Staphylococcus aureus of the ethanol extract (0.01 g/mL) of Centella asiatica.
Figure 2 is a photograph showing the results of an antibacterial test against Escherichia coli and Staphylococcus aureus of the ethanol extract of Centella asiatica (0.02 g/mL).
Figure 3 is a photograph showing the results of an antibacterial test against Escherichia coli and Staphylococcus aureus of the ethanol extract of Centella asiatica (0.03 g/mL).
Figure 4 is a photograph showing the results of an antibacterial test against Escherichia coli and Staphylococcus aureus of the ethanol extract of Centella asiatica (0.05 g/mL).
Figure 5 is a photograph showing the results of an antibacterial test against Pseudomonas aeruginosa and Klebsiella pneumoniae of the ethanol extract (0.05 g/mL) of Centella asiatica.
Figure 6 is a photograph showing the results of an antibacterial test against Pseudomonas aeruginosa and Klebsiella pneumoniae of an ethanol extract (0.03 g/mL) of Centella asiatica.
Figure 7 is a photograph showing the results of an antibacterial test against Pseudomonas aeruginosa and Klebsiella pneumoniae of an ethanol extract (0.02 g/mL) of Centella asiatica.
Figure 8 is a photograph showing the results of an antibacterial test against Pseudomonas aeruginosa and Klebsiella pneumoniae of the ethanol extract (0.01 g/mL) of Centella asiatica.
이하에서, 첨부된 도면을 참조하여 실시예들을 상세하게 설명한다. 그러나, 실시예들에는 다양한 변경이 가해질 수 있어서 특허출원의 권리 범위가 이러한 실시예들에 의해 제한되거나 한정되는 것은 아니다. 실시예들에 대한 모든 변경, 균등물 내지 대체물이 권리 범위에 포함되는 것으로 이해되어야 한다.Hereinafter, embodiments will be described in detail with reference to the attached drawings. However, various changes can be made to the embodiments, so the scope of the patent application is not limited or limited by these embodiments. It should be understood that all changes, equivalents, or substitutes for the embodiments are included in the scope of rights.
실시예에서 사용한 용어는 단지 설명을 목적으로 사용된 것으로, 한정하려는 의도로 해석되어서는 안된다. 단수의 표현은 문맥상 명백하게 다르게 뜻하지 않는 한, 복수의 표현을 포함한다. 본 명세서에서, "포함하다" 또는 "가지다" 등의 용어는 명세서 상에 기재된 특징, 숫자, 단계, 동작, 구성요소, 부품 또는 이들을 조합한 것이 존재함을 지정하려는 것이지, 하나 또는 그 이상의 다른 특징들이나 숫자, 단계, 동작, 구성요소, 부품 또는 이들을 조합한 것들의 존재 또는 부가 가능성을 미리 배제하지 않는 것으로 이해되어야 한다.The terms used in the examples are for descriptive purposes only and should not be construed as limiting. Singular expressions include plural expressions unless the context clearly dictates otherwise. In this specification, terms such as “comprise” or “have” are intended to designate the presence of features, numbers, steps, operations, components, parts, or combinations thereof described in the specification, but are not intended to indicate the presence of one or more other features. It should be understood that this does not exclude in advance the possibility of the existence or addition of elements, numbers, steps, operations, components, parts, or combinations thereof.
다르게 정의되지 않는 한, 기술적이거나 과학적인 용어를 포함해서 여기서 사용되는 모든 용어들은 실시예가 속하는 기술 분야에서 통상의 지식을 가진 자에 의해 일반적으로 이해되는 것과 동일한 의미를 가지고 있다. 일반적으로 사용되는 사전에 정의되어 있는 것과 같은 용어들은 관련 기술의 문맥 상 가지는 의미와 일치하는 의미를 가지는 것으로 해석되어야 하며, 본 출원에서 명백하게 정의하지 않는 한, 이상적이거나 과도하게 형식적인 의미로 해석되지 않는다.Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as generally understood by a person of ordinary skill in the technical field to which the embodiments belong. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning in the context of the related technology, and unless explicitly defined in the present application, should not be interpreted in an ideal or excessively formal sense. No.
또한, 첨부 도면을 참조하여 설명함에 있어, 도면 부호에 관계없이 동일한 구성 요소는 동일한 참조부호를 부여하고 이에 대한 중복되는 설명은 생략하기로 한다. 실시예를 설명함에 있어서 관련된 공지 기술에 대한 구체적인 설명이 실시예의 요지를 불필요하게 흐릴 수 있다고 판단되는 경우 그 상세한 설명을 생략한다.In addition, when describing with reference to the accompanying drawings, identical components will be assigned the same reference numerals regardless of the reference numerals, and overlapping descriptions thereof will be omitted. In describing the embodiments, if it is determined that detailed descriptions of related known technologies may unnecessarily obscure the gist of the embodiments, the detailed descriptions are omitted.
또한, 실시 예의 구성 요소를 설명하는 데 있어서, 제 1, 제 2, A, B, (a), (b) 등의 용어를 사용할 수 있다. 이러한 용어는 그 구성 요소를 다른 구성 요소와 구별하기 위한 것일 뿐, 그 용어에 의해 해당 구성 요소의 본질이나 차례 또는 순서 등이 한정되지 않는다. 어떤 구성 요소가 다른 구성요소에 "연결", "결합" 또는 "접속"된다고 기재된 경우, 그 구성 요소는 그 다른 구성요소에 직접적으로 연결되거나 접속될 수 있지만, 각 구성 요소 사이에 또 다른 구성 요소가 "연결", "결합" 또는 "접속"될 수도 있다고 이해되어야 할 것이다.Additionally, in describing the components of the embodiment, terms such as first, second, A, B, (a), and (b) may be used. These terms are only used to distinguish the component from other components, and the nature, sequence, or order of the component is not limited by the term. When a component is described as being "connected," "coupled," or "connected" to another component, that component may be directly connected or connected to that other component, but there is no need for another component between each component. It should be understood that may be “connected,” “combined,” or “connected.”
어느 하나의 실시 예에 포함된 구성요소와, 공통적인 기능을 포함하는 구성요소는, 다른 실시 예에서 동일한 명칭을 사용하여 설명하기로 한다. 반대되는 기재가 없는 이상, 어느 하나의 실시 예에 기재한 설명은 다른 실시 예에도 적용될 수 있으며, 중복되는 범위에서 구체적인 설명은 생략하기로 한다.Components included in one embodiment and components including common functions will be described using the same names in other embodiments. Unless stated to the contrary, the description given in one embodiment may be applied to other embodiments, and detailed description will be omitted to the extent of overlap.
본 발명의 일 측면은, 긴병꽃풀(Glechoma grandis (A. Gray) Kuprian) 지상부 추출물을 유효성분으로 포함하는, 항균제 조성물을 제공한다.One aspect of the present invention provides an antibacterial composition comprising an extract of the aerial parts of Glechoma grandis (A. Gray) Kuprian as an active ingredient.
본 발명에 따른 항균제 조성물은, 긴병꽃풀 지상부 추출물을 포함하는 천연 물질 기반의 항균제 조성물로서, 우수한 항균력을 가지며 인체 안전성이 확보되는 효과가 있다. The antibacterial composition according to the present invention is a natural substance-based antibacterial composition containing an extract of the above-ground parts of Centella asiatica, and has excellent antibacterial activity and has the effect of ensuring human safety.
상기 긴병꽃풀(Glechoma grandis (A. Gray) Kuprian) 지상부 추출물은, 긴병꽃풀의 잎, 줄기를 비롯한 지상부 영역의 추출물을 의미할 수 있다.The extract of the aerial part of Glechoma grandis (A. Gray) Kuprian may refer to the extract of the above-ground area, including the leaves and stems of the plant.
상기 긴병꽃풀 지상부 추출물은, 피부자극성 및 안점막 자극성이 없어 피부 안전성을 확보할 수 있다.The above-mentioned extract of the above-mentioned part of Centella asiatica does not cause skin irritation or eye mucous membrane irritation, thereby ensuring skin safety.
또한, 단회경구투여(최고 용량 2,000 mg/kg Body weight) 및 단회경피투여(최고 용량 2,000 mg/kg Body weight) 시 독성이 없음이 입증되어, 최고 용량을 초과하지 않는 범위에서 경구투여 및 경피투여될 수 있다. 즉, 상기 긴병꽃풀 지상부 추출물은, 경구용 제제 또는 피부 외용제의 형태로 제조되어 인체에 경구투여 또는 경피투여될 수 있다.In addition, it has been proven to be non-toxic when administered via single oral administration (maximum dose 2,000 mg/kg body weight) and single transdermal administration (maximum dose 2,000 mg/kg body weight), so it can be administered orally and transdermally within the range of not exceeding the maximum dose. It can be. In other words, the extract of the above-mentioned parts of Centella asiatica can be prepared in the form of an oral preparation or topical skin preparation and administered orally or transdermally to the human body.
일 실시형태에 따르면, 상기 항균제 조성물 중, 상기 긴병꽃풀 지상부 추출물의 농도는, 0.01 g/ml 내지 0.05 g/ml인 것일 수 있다.According to one embodiment, in the antibacterial composition, the concentration of the aerial part extract of Centella asiatica may be 0.01 g/ml to 0.05 g/ml.
상기 긴병꽃풀 지상부 추출물의 농도 범위는, 대장균, 황색포도상구균, 폐렴간균 및 녹농균에 대한 항균력을 모두 확보하면서, 항균제 제품에 적용시 제조 용이성 및 경제성을 확보할 수 있는 농도 범위일 수 있다.The concentration range of the above-mentioned extract of the aerial parts of Centella asiatica longifolia may be a concentration range that secures antibacterial activity against Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, and Pseudomonas aeruginosa while ensuring ease of manufacture and economic feasibility when applied to antibacterial products.
일 실시형태에 따르면, 상기 긴병꽃풀 지상부 추출물은, 폴리페놀 5 중량% 내지 10 중량%를 포함하는 것일 수 있다. 바람직하게는, 폴리페놀 7 중량% 내지 9 중량%를 포함하는 것일 수 있다.According to one embodiment, the extract of the above-ground parts of Centella asiatica may contain 5% to 10% by weight of polyphenol. Preferably, it may contain 7% to 9% by weight of polyphenol.
폴리페놀은 다수의 페놀 작용기를 가진 고분자로, 우리 몸에 있는 활성산소를 해가 없는 물질로 바꿔주는 항산화물질 중 하나로 알려져 있으며, 그 종류는 수천 가지가 넘는다. 잘 알려진 예로는 녹차에 든 카테킨, 포도주의 레스베라트롤, 사과와 양파의 쿼세틴 등이 있으며, 과일에 많이 함유된 플라보노이드와 콩에 많은 이소플라본도 폴리페놀의 일종이다. 또한, 폴리페놀은 항균 활성을 나타내는 것으로 알려져 있다.Polyphenol is a polymer with multiple phenol functional groups, and is known as one of the antioxidant substances that converts free radicals in our body into harmless substances, and there are over thousands of types. Well-known examples include catechin in green tea, resveratrol in wine, and quercetin in apples and onions. Flavonoids, which are abundant in fruits, and isoflavones, which are abundant in beans, are also types of polyphenols. Additionally, polyphenols are known to exhibit antibacterial activity.
상기 긴병꽃풀 지상부 추출물은, 유효성분으로서 폴리페놀을 추출물 전체 중 5 중량% 내지 10 중량%, 바람직하게는 7 중량% 내지 9중량% 포함함으로써, 항균 활성이 향상되는 효과가 있다.The extract of the aerial parts of Centella asiatica has the effect of improving antibacterial activity by containing polyphenol as an active ingredient in an amount of 5% to 10% by weight, preferably 7% to 9% by weight, based on the total extract.
일 실시형태에 따르면, 상기 긴병꽃풀 지상부 추출물은, 물, 탄소수 1 내지 4의 저급 알코올, 아세톤, 에틸아세테이트, 클로로포름, 1,3-부틸렌글리콜, n-헥산, 디에틸에테르, 부틸아세테이트 및 디클로로메탄으로 이루어진 군에서 선택되는 하나 이상을 추출 용매로 하여 추출된 것일 수 있다.According to one embodiment, the extract of the above-ground part of Centella asiatica is composed of water, lower alcohol having 1 to 4 carbon atoms, acetone, ethyl acetate, chloroform, 1,3-butylene glycol, n-hexane, diethyl ether, butyl acetate, and It may be extracted using at least one selected from the group consisting of dichloromethane as an extraction solvent.
일 실시형태에 따르면, 상기 저급 알코올은, 에탄올인 것일 수 있다.According to one embodiment, the lower alcohol may be ethanol.
바람직하게는, 상기 저급 알코올은, 50% (v/v) 내지 90% (v/v) 에탄올일 수 있고, 더욱 바람직하게는, 60% (v/v) 내지 80% (v/v) 에탄올인 것일 수 있다.Preferably, the lower alcohol may be 50% (v/v) to 90% (v/v) ethanol, more preferably 60% (v/v) to 80% (v/v) ethanol. It may be.
상기 에탄올의 농도 범위는, 상기 긴병꽃풀 지상부 추출물 내 유효성분(폴리페놀 포함)의 추출 효율을 최대화할 수 있는 범위일 수 있다.The concentration range of the ethanol may be a range that can maximize the extraction efficiency of the active ingredients (including polyphenols) in the extract of the aerial parts of Centella asiatica.
일 실시형태에 따르면, 상기 추출은, 20 ℃ 내지 30 ℃의 온도에서 24 내지 48 시간 동안 수행되는 것일 수 있다.According to one embodiment, the extraction may be performed at a temperature of 20°C to 30°C for 24 to 48 hours.
일 실시형태에 따르면, 상기 추출은, 0.1 MPa 내지 0.2 MPa 압력 조건에서 수행되는 것일 수 있다.According to one embodiment, the extraction may be performed under pressure conditions of 0.1 MPa to 0.2 MPa.
상기 온도, 시간 및 압력 조건 범위를 벗어날 경우, 상기 긴병꽃풀 추출물 내 유효성분이 충분히 추출되지 않거나 유효성분이 파괴될 수 있다.If the temperature, time, and pressure conditions are outside the range, the active ingredients in the Centella asiatica extract may not be sufficiently extracted or the active ingredients may be destroyed.
일 실시형태에 따르면, 폐렴간균(Klebsiella pneumoniae), 녹농균(Pseudomonas aeruginosa), 포도상구균(Staphylococcus aureus) 및 대장균(Escherichia coli) 중 적어도 하나 이상의 균주에 대하여 항균 활성을 갖는 것일 수 있다.According to one embodiment, it may have antibacterial activity against at least one strain of Klebsiella pneumoniae , Pseudomonas aeruginosa , Staphylococcus aureus , and Escherichia coli .
일 실시형태에 따르면, 상기 항균제 조성물은, 폐렴간균(Klebsiella pneumoniae), 녹농균(Pseudomonas aeruginosa), 포도상구균(Staphylococcus aureus) 및 대장균(Escherichia coli) 중 적어도 하나 이상의 균주에 대하여, 99.9% 이상의 균감소율을 나타내는 것일 수 있다.According to one embodiment, the antibacterial composition has a bacterial reduction rate of 99.9% or more against at least one strain of Klebsiella pneumoniae , Pseudomonas aeruginosa , Staphylococcus aureus , and Escherichia coli. It may represent.
본 발명에 따른 항균제 조성물은, 폐렴간균(Klebsiella pneumoniae), 녹농균(Pseudomonas aeruginosa), 포도상구균(Staphylococcus aureus) 및 대장균(Escherichia coli) 중 적어도 하나 이상의 균주에 대하여, 99.9% 이상의 균감소율을 나타냄으로써 우수한 항균 활성을 가지며, 다양한 항균제품에 적용 가능한 특징을 갖는다.The antibacterial composition according to the present invention is excellent by showing a bacterial reduction rate of more than 99.9% against at least one strain of Klebsiella pneumoniae, Pseudomonas aeruginosa , Staphylococcus aureus , and Escherichia coli. It has antibacterial activity and can be applied to a variety of antibacterial products.
일 실시형태에 따르면, 상기 항균제 조성물은, 용액, 현탁액, 유탁액, 페이스트, 겔, 크림, 로션, 파우더, 비누, 계면활성제 함유 클렌징, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션 및 스프레이로 이루어진 군에서 선택되는 하나 이상의 제형을 포함하는 것일 수 있다.According to one embodiment, the antibacterial composition consists of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, and spray. It may include one or more formulations selected from the group.
본 발명의 다른 측면은, 상기 항균제 조성물을 포함하는, 항균용 피부 외용제를 제공한다. Another aspect of the present invention provides an antibacterial external skin preparation comprising the antibacterial composition.
본 발명에 따른 항균용 피부 외용제는, 피부 자극성 및 안점막 자극성이 없어 피부 안전성이 우수한 효과를 갖는다.The antibacterial external skin preparation according to the present invention has excellent skin safety because it does not cause skin irritation or ocular mucous membrane irritation.
일 실시형태에 따르면, 상기 항균용 피부 외용제 중, 상기 항균제 조성물은 1 중량% 내지 5 중량%로 포함되는 것일 수 있다.According to one embodiment, in the antibacterial skin external preparation, the antibacterial agent composition may be included in an amount of 1% to 5% by weight.
상기 항균제 조성물의 함량이 상기 범위 미만일 경우 도포 부위에 항균 효과가 충분히 나타나지 않을 수 있고, 상기 범위를 초과할 경우 도포력 또는 피부 흡수력이 저하될 수 있다. If the content of the antibacterial composition is less than the above range, the antibacterial effect may not be sufficiently shown at the application site, and if it exceeds the above range, the application power or skin absorption power may be reduced.
상기 피부 외용제는, 일반적으로 외용에 사용하는 조성물 전반을 포괄하는 개념으로, 기초 화장료, 메이크업 화장료, 모발용 화장료, 면도용 화장료 등 각종 화장료를 포함하는 화장료 조성물이나 연고제 등의 다양한 의약품 내지 의약외품 등으로 폭넓게 적용 가능한 것을 의미할 수 있다. 예를 들어, 상기 의약외품은 소독청결제, 샤워폼, 가그린, 물티슈, 세제비누, 핸드워시, 가습기 충진제, 마스크, 연고제, 코팅제 또는 필터충진제일 수 있다.The skin external preparation is a concept that generally encompasses all compositions used for external use, and is broadly divided into cosmetic compositions containing various cosmetics such as basic cosmetics, makeup cosmetics, hair cosmetics, and shaving cosmetics, as well as various pharmaceuticals and quasi-drugs such as ointments. It can mean applicable. For example, the quasi-drug may be a disinfectant cleaner, shower foam, garnish, wet tissue, detergent soap, hand wash, humidifier filler, mask, ointment, coating agent, or filter filler.
상기 피부 외용제 조성물은 유효성분 외에 사용용도 및 외용제 조성물의 성질에 따라 통상 외용제에 배합되고 있는 성분, 구체적으로 보습제, 자외선 흡수제, 비타민류, 동식 물 추출성분, 소화제, 미백제, 혈관확장제, 수렴제, 청량제, 호르몬제 등을 추가로 배합한 것일 수 있다.In addition to the active ingredients, the composition for external use for skin includes ingredients that are usually mixed in external preparations depending on the intended use and the properties of the external preparation composition, specifically, moisturizers, ultraviolet absorbers, vitamins, animal and vegetable extracts, digestive agents, whitening agents, vasodilators, astringents, and refreshing agents. , may be additionally mixed with hormones, etc.
상기 피부 외용제 조성물의 제형은 사용용도 및 외용제 조성물의 성질에 따라 적절한 형태를 취할 수 있으며, 구체적으로 수용액계, 가용화계, 유화계, 유액계, 겔계, 페이스트계, 연고계, 에어졸계, 물-기름 2층계, 물-기름-분말 3층계일 수 있으며, 상기 제형에 의해 본 발명의 외용제의 제형 및 형태가 제한되는 것은 아니다.The formulation of the skin external preparation composition may take an appropriate form depending on the intended use and the nature of the external preparation composition, specifically, aqueous solution, solubilization, emulsification, emulsion, gel, paste, ointment, aerosol, water-based. It may be a two-layer oil system or a three-layer water-oil-powder system, and the formulation and form of the external preparation of the present invention are not limited by the above formulation.
또한, 상기 피부 외용제는 유효성분을 피부 조직으로 침투 또는 이행시키기 위해서 필요한 기제를 추가로 포함할 수 있다.In addition, the skin external preparation may additionally contain a mechanism necessary to penetrate or transfer the active ingredient into skin tissue.
일 실시형태에 따르면, 상기 항균용 피부 외용제는, 용액제, 현탁제, 스프레이제, 패취제, 패드제, 크림제, 연고제 또는 겔제의 제형으로 제조될 수 있다.According to one embodiment, the antibacterial external skin preparation may be prepared in the form of a solution, suspension, spray, patch, pad, cream, ointment, or gel.
용액제는 유효 화합물 이외에 통상의 부형제, 예를 들면 용매, 가용화제 및 유화제, 예를 들면 물, 에틸알코올, 이소프로필알코올, 에틸카보네이트, 에틸아세테이트, 벤질알코올, 벤질벤조에이트, 프로필렌글리콜, 1,3-부틸렌글리콜, 디메틸포름아미드, 오일, 특히 면실유, 낙화생유, 동백유, 알로에베라, 글리세린, 천연옥수수 배종유, 올리브유, 피마자유, 아몬드유 및 참깨유, 글리세롤, 글리세롤 포름알코올, 테티라히드로푸르푸릴 알코올, 폴리에틸렌 글리콜 및 소르비탄의 지방산 에스테르, 또는 이들 물질의 혼합물을 포함할 수 있으며, 보존제로서 메틸 또는 프로필-p-히드록시벤조에이트 또는 솔브산 등을 포함할 수 있다.In addition to the active compound, the solution contains common excipients, such as solvents, solubilizers and emulsifiers, such as water, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1, 3-Butylene glycol, dimethylformamide, oils, especially cottonseed oil, peanut oil, camellia oil, aloe vera, glycerin, natural corn pear seed oil, olive oil, castor oil, almond oil and sesame oil, glycerol, glycerol formal alcohol, tetrahydro. It may contain furfuryl alcohol, fatty acid esters of polyethylene glycol and sorbitan, or a mixture of these substances, and may contain methyl or propyl-p-hydroxybenzoate or sorbic acid as a preservative.
현탁제는 유효 화합물 이외에 통상의 부형제, 예를 들면 액상 희석제 (예: 물, 에틸알코올 및 프로필렌 글리콜, 폴리에틸렌 글리콜), 및 현탁제 (예: 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨, 소르비탄 에스테르, 셀룰로스 유도체 및 수소화 식용유지), 미세 결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 한천 및 트라가칸트, 에틸올레이트와 같은 주사 가능한 에스테르 또는 이들 물질의 혼합물을 함유할 수 있다.Suspending agents include, in addition to the active compound, conventional excipients, such as liquid diluents (e.g. water, ethyl alcohol and propylene glycol, polyethylene glycol), and suspending agents (e.g. ethoxylated isostearyl alcohol, polyoxyethylene sorbitol, sorbitan). esters, cellulose derivatives and hydrogenated edible oils), microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and tragacanth, injectable esters such as ethyl oleate or mixtures of these substances.
연고제, 크림제 및 겔제는 유효 화합물 이외에 통상의 부형제, 예를 들면 동물성 및 식물성 지방, 왁스 파라핀, 전분, 타르가칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 규산, 활석 및 산화 아연 또는 이들 물질의 혼합물을 함유할 수 있다.Ointments, creams and gels contain, in addition to the active compound, customary excipients, such as animal and vegetable fats, wax paraffin, starch, targacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silicic acid, talc and zinc oxide or these substances. It may contain a mixture of
본 발명의 또 다른 측면은, 상기 항균제 조성물을 포함하는, 식품 첨가물, 의약품, 수의약품 또는 화장품 조성물을 제공한다.Another aspect of the present invention provides a food additive, pharmaceutical, veterinary drug, or cosmetic composition comprising the antibacterial agent composition.
상기 식품 첨가물, 의약품, 수의약품 또는 화장품 조성물은, 분말, 과립, 캡슐, 환제, 정제, 츄잉검과 같은 고체 형태 또는 시럽, 용액, 현탁액과 같은 액체 형태를 비롯한 경구 투여에 적합한 임의의 형태로 제형화될 수 있다. 또한, 크림, 연고, 겔, 밀크, 페이스트, 유제, 형청, 마스크, 향유, 체액, 스프레이, 좌제, 질 좌약, 경피 흡수제 패치, 치약, 치주젤, 구강 청결제 등과 같은 형태로 제형화될 수 있다.The food additive, drug, veterinary drug, or cosmetic composition may be formulated in any form suitable for oral administration, including solid form such as powder, granule, capsule, pill, tablet, chewing gum, or liquid form such as syrup, solution, or suspension. It can get angry. Additionally, it can be formulated in forms such as cream, ointment, gel, milk, paste, emulsion, cream, mask, balm, body fluid, spray, suppository, vaginal suppository, transdermal absorbent patch, toothpaste, periodontal gel, mouthwash, etc.
일 실시형태에 따르면, 상기 항균제 조성물은, 경구 또는 국소 투여용으로 제형화될 수 있다. According to one embodiment, the antibacterial composition may be formulated for oral or topical administration.
상기 항균제 조성물을 포함하는 의약품 또는 수의약품의 경우, 상기 항균제 조성물이 치료학적 유효량으로 포함 또는 투여된다.In the case of a pharmaceutical or veterinary drug containing the antibacterial agent composition, the antibacterial agent composition is contained or administered in a therapeutically effective amount.
본 발명의 다른 측면은, 긴병꽃풀의 지상부를 건조 및 분쇄하여 긴병꽃풀 지상부 건조물을 제조하는 단계; 상기 긴병꽃풀 지상부 건조물을 용매 추출하여 긴병꽃풀 지상부 추출물을 얻는 단계; 상기 긴병꽃풀 지상부 추출물을 여과한 후 감압 농축하여 용매를 제거하는 단계; 및 상기 용매가 제거된 긴병꽃풀 지상부 추출물을 동결 건조하는 단계;를 포함하는, 긴병꽃풀 지상부 추출물의 제조방법을 제공한다.Another aspect of the present invention includes the steps of drying and pulverizing the above-ground parts of the long-term Callaena longifolia to produce a dried above-ground part of the long-distance Calliae longifolia; Obtaining an extract of the above-mentioned part of the above-mentioned part of the above-mentioned part by solvent extraction; Filtering the above-mentioned extract of the aerial parts of Centella asiatica and then concentrating under reduced pressure to remove the solvent; And a step of freeze-drying the extract of the above-mentioned part of the above-mentioned part of the above-mentioned part of the above-mentioned part of the above-described part of the extract from which the solvent has been removed.
상기 긴병꽃풀의 지상부는, 긴병꽃풀의 잎 및 줄기 중 적어도 하나를 포함할 수 있다. The above-ground part of the plant may include at least one of the leaves and stems of the plant.
일 실시형태에 따르면, 상기 긴병꽃풀 지상부 추출물을 얻는 단계는, 상기 긴병꽃풀 지상부 건조물을 20 ℃ 내지 30 ℃ 온도로 24 시간 내지 48 시간 동안 용매에 침지시킨 후, 상등액과 침전물을 분리하여 긴병꽃풀 지상부 추출물을 수득하는 것일 수 있다.According to one embodiment, the step of obtaining the extract of the above-mentioned part of Centella asiatica includes immersing the dried above-mentioned part of Centella asiatica in a solvent at a temperature of 20° C. to 30° C. for 24 to 48 hours, then separating the supernatant and the precipitate and It may be to obtain an extract of the aerial parts of Centella asiatica.
여기서, 상기 상등액과 침전물을 분리하는 과정은 반복 수행될 수 있다.Here, the process of separating the supernatant and precipitate may be repeated.
일 실시형태에 따르면, 상기 긴병꽃풀 지상부 추출물을 얻는 단계는, 1회 이상 수행될 수 있고, 바람직하게는, 3회 이상 반복 수행될 수 있다. 이는, 긴병꽃풀 지상부 내 포함된 유효성분을 충분히 추출하기 위함일 수 있다.According to one embodiment, the step of obtaining the extract of the above-mentioned aerial parts of Centella asiatica may be performed one or more times, and preferably, may be repeated three or more times. This may be to sufficiently extract the active ingredients contained in the above-ground parts of the long-distance plant.
이하, 실시예 및 비교예에 의하여 본 발명을 더욱 상세히 설명하고자 한다.Hereinafter, the present invention will be described in more detail through examples and comparative examples.
단, 하기 실시예는 본 발명을 예시하기 위한 것일 뿐, 본 발명의 내용이 하기 실시예에 한정되는 것은 아니다.However, the following examples are only for illustrating the present invention, and the content of the present invention is not limited to the following examples.
<실시예 1> 긴병꽃풀 지상부 추출물의 제조<Example 1> Preparation of extract of aerial parts of Centella asiatica
긴병꽃풀의 줄기와 잎 5 kg을 채취하여 열풍건조 또는 자연 음지 건조 또는 동결 건조후 분쇄하였다. 분쇄한 시료에 시료 중량의 10배 양의 에탄올 70% (v/v)을 가하여 실온에서 약 24 시간 침지한 후 상등액과 침전물을 분리하여 동일한 방법으로 3회 반복 추출하였다. 각 시료 추출물은 여과지(Whatman No.2)를 이용하여 여과한 후 EYELA 증발기(evaporator)로 감압 농축하여 용매를 제거 후 동결 건조하였으며, 이 시료를 - 20 ℃ 온도에서 보관하였다. 이때, 긴병꽃풀은 오산 일원에서 채취하였다.5 kg of stems and leaves of Centella asiatica were collected, dried in hot air, naturally shaded, or freeze-dried, and then ground. 70% (v/v) ethanol in an amount 10 times the weight of the sample was added to the ground sample and soaked at room temperature for about 24 hours. The supernatant and precipitate were separated and extracted three times in the same manner. Each sample extract was filtered using filter paper (Whatman No. 2) and then concentrated under reduced pressure with an EYELA evaporator to remove the solvent and freeze-dried. The sample was stored at -20°C. At this time, Byeongbyeol was collected from the Osan area.
<실시예 2> 긴병꽃풀 지상부 추출물을 포함하는 피부 외용제 제조<Example 2> Preparation of topical skin preparation containing extract of aerial parts of Centella asiatica
실시예 1의 긴병꽃풀 지상부 추출물에 정제수, 에탄올 및 레몬 오일 등을 첨가하여 피부 외용제를 제조하였다.An external skin preparation was prepared by adding purified water, ethanol, and lemon oil to the extract of the aerial parts of Centella asiatica in Example 1.
<실험예 1> 긴병꽃풀 지상부 추출물의 대장균 및 황색포도상구균에 대한 항균력 평가<Experimental Example 1> Evaluation of antibacterial activity against Escherichia coli and Staphylococcus aureus of extracts from the aerial parts of Centella asiatica
실시예 1의 방법으로 제조된 긴병꽃풀 지상부 추출물의 농도 및 균주별 항균력을 각각 평가하였다. The concentration and antibacterial activity of each strain of the extract of the aerial parts of Centella asiatica produced by the method of Example 1 were evaluated.
시험 방법 및 결과를 표 1내지 표 4 및 도 1 내지 도 4에 나타내었다.Test methods and results are shown in Tables 1 to 4 and Figures 1 to 4.
1) 긴병꽃풀 에탄올 추출물(0.01 g/mL)에 대한 항균력 평가1) Evaluation of antibacterial activity of ethanol extract of Centella asiatica (0.01 g/mL)
(CFU/mL)Initial concentration
(CFU/mL)
(CFU/mL)
Concentration after 24 hours
(CFU/mL)
(%)decline rate
(%)
대장균Antibacterial test:
E. coli
(0.01g/ml)Centella asiatica ethanol extract
(0.01g/ml)
황색포도상구균Antibacterial test:
Staphylococcus aureus
(0.01g/ml)Centella asiatica ethanol extract
(0.01g/ml)
* CFU : Colony Forming Unit* 접종원 농도(CFU/mL) : 대장균 : 9.0 x 106, 황색포도상구균 : 3.7 x 106 * CFU: Colony Forming Unit* Inoculant concentration (CFU/mL): Escherichia coli: 9.0 x 10 6 , Staphylococcus aureus: 3.7 x 10 6
* 사용균주 : Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 6538* Strains used: Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 6538
* 반응시간 : 24시간* Response time: 24 hours
도 1은, 긴병꽃풀 에탄올 추출물(0.01 g/mL)의 대장균 및 황색포도상구균에 대한 항균시험 결과를 나타낸 사진이다.Figure 1 is a photograph showing the results of an antibacterial test against Escherichia coli and Staphylococcus aureus of the ethanol extract (0.01 g/mL) of Centella asiatica.
표 1 및 도 1을 참조하면, 긴병꽃풀 에탄올 추출물(0.01 g/mL)은 대장균 및 황색포도상구균에 대해 99.9%의 항균활성을 나타내는 것을 확인할 수 있다.Referring to Table 1 and Figure 1, it can be seen that the ethanol extract of Centella asiatica (0.01 g/mL) exhibits 99.9% antibacterial activity against Escherichia coli and Staphylococcus aureus.
2) 긴병꽃풀 에탄올 추출물(0.02 g/mL)에 대한 항균력 평가2) Evaluation of antibacterial activity of ethanol extract of Centella asiatica (0.02 g/mL)
(CFU/mL)Initial concentration
(CFU/mL)
(CFU/mL)
Concentration after 24 hours
(CFU/mL)
(%)decline rate
(%)
대장균Antibacterial test:
E. coli
(0.02g/ml)Centella asiatica ethanol extract
(0.02g/ml)
황색포도상구균Antibacterial test:
Staphylococcus aureus
(0.02g/ml)Centella asiatica ethanol extract
(0.02g/ml)
* CFU : Colony Forming Unit* 접종원 농도(CFU/mL) : 대장균 : 9.0 x 106, 황색포도상구균 : 3.7 x 106 * CFU: Colony Forming Unit* Inoculant concentration (CFU/mL): Escherichia coli: 9.0 x 10 6 , Staphylococcus aureus: 3.7 x 10 6
* 사용균주 : Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 6538* Strains used: Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 6538
* 반응시간 : 24시간* Response time: 24 hours
도 2는, 긴병꽃풀 에탄올 추출물(0.02 g/mL)의 대장균 및 황색포도상구균에 대한 항균시험 결과를 나타낸 사진이다.Figure 2 is a photograph showing the results of an antibacterial test against Escherichia coli and Staphylococcus aureus of the ethanol extract of Centella asiatica (0.02 g/mL).
표 2 및 도 2를 참조하면, 긴병꽃풀 에탄올 추출물(0.02 g/mL)은 대장균 및 황색포도상구균에 대해 99.9%의 항균활성을 나타내는 것을 확인할 수 있다.Referring to Table 2 and Figure 2, it can be seen that the ethanol extract of Centella asiatica (0.02 g/mL) exhibits 99.9% antibacterial activity against Escherichia coli and Staphylococcus aureus.
3) 긴병꽃풀 에탄올 추출물(0.03 g/mL)에 대한 항균력 평가3) Evaluation of antibacterial activity of ethanol extract of Centella asiatica (0.03 g/mL)
(CFU/mL)Initial concentration
(CFU/mL)
(CFU/mL)
Concentration after 24 hours
(CFU/mL)
(%)decline rate
(%)
대장균Antibacterial test:
E. coli
(0.03g/ml)Centella asiatica ethanol extract
(0.03g/ml)
황색포도상구균Antibacterial test:
Staphylococcus aureus
(0.03g/ml)Centella asiatica ethanol extract
(0.03g/ml)
* CFU : Colony Forming Unit* 접종원 농도(CFU/mL) : 대장균 : 9.0 x 106, 황색포도상구균 : 3.7 x 106 * CFU: Colony Forming Unit* Inoculant concentration (CFU/mL): Escherichia coli: 9.0 x 10 6 , Staphylococcus aureus: 3.7 x 10 6
* 사용균주 : Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 6538* Strains used: Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 6538
* 반응시간 : 24시간* Response time: 24 hours
도 3은, 긴병꽃풀 에탄올 추출물(0.03 g/mL)의 대장균 및 황색포도상구균에 대한 항균시험 결과를 나타낸 사진이다.Figure 3 is a photograph showing the results of an antibacterial test against Escherichia coli and Staphylococcus aureus of the ethanol extract of Centella asiatica (0.03 g/mL).
표 3 및 도 3을 참조하면, 긴병꽃풀 에탄올 추출물(0.03 g/mL)은 대장균 및 황색포도상구균에 대해 99.9%의 항균활성을 나타내는 것을 확인할 수 있다.Referring to Table 3 and Figure 3, it can be seen that the ethanol extract of Centella asiatica (0.03 g/mL) exhibits 99.9% antibacterial activity against Escherichia coli and Staphylococcus aureus.
4) 긴병꽃풀 에탄올 추출물(0.05 g/mL)에 대한 항균력 평가4) Evaluation of antibacterial activity of ethanol extract of Centella asiatica (0.05 g/mL)
(CFU/mL)Initial concentration
(CFU/mL)
(CFU/mL)
Concentration after 24 hours
(CFU/mL)
(%)decline rate
(%)
대장균Antibacterial test:
E. coli
(0.05g/ml)Centella asiatica ethanol extract
(0.05g/ml)
황색포도상구균Antibacterial test:
Staphylococcus aureus
(0.05g/ml)Centella asiatica ethanol extract
(0.05g/ml)
* CFU : Colony Forming Unit* 접종원 농도(CFU/mL) : 대장균 : 9.0 x 106, 황색포도상구균 : 3.7 x 106 * CFU: Colony Forming Unit* Inoculant concentration (CFU/mL): Escherichia coli: 9.0 x 10 6 , Staphylococcus aureus: 3.7 x 10 6
* 사용균주 : Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 6538* Strains used: Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 6538
* 반응시간 : 24시간* Response time: 24 hours
도 4는, 긴병꽃풀 에탄올 추출물(0.05 g/mL)의 대장균 및 황색포도상구균에 대한 항균시험 결과를 나타낸 사진이다.Figure 4 is a photograph showing the results of an antibacterial test against Escherichia coli and Staphylococcus aureus of the ethanol extract of Centella asiatica (0.05 g/mL).
표 4 및 도 4를 참조하면, 긴병꽃풀 에탄올 추출물(0.05 g/mL)은 대장균 및 황색포도상구균에 대해 99.9%의 항균활성을 나타내는 것을 확인할 수 있다.Referring to Table 4 and Figure 4, it can be seen that the ethanol extract of Centella asiatica (0.05 g/mL) exhibits 99.9% antibacterial activity against Escherichia coli and Staphylococcus aureus.
<실험예 2> 긴병꽃풀 지상부 추출물의 녹농균 및 폐렴간균에 대한 항균력 평가<Experimental Example 2> Evaluation of antibacterial activity against Pseudomonas aeruginosa and Klebsiella pneumoniae of extracts from the aerial parts of Centella asiatica
실시예 1의 방법으로 제조된 긴병꽃풀 지상부 추출물의 농도 및 균주별 항균력을 각각 평가하였다. The concentration and antibacterial activity of each strain of the extract of the aerial parts of Centella asiatica produced by the method of Example 1 were evaluated.
시험 방법 및 결과를 표 5내지 표 8 및 도 5 내지 도 8에 나타내었다.Test methods and results are shown in Tables 5 to 8 and Figures 5 to 8.
1) 긴병꽃풀 에탄올 추출물(0.05 g/mL)에 대한 항균력 평가1) Evaluation of antibacterial activity of ethanol extract of Centella asiatica (0.05 g/mL)
(CFU/mL)Initial concentration
(CFU/mL)
(CFU/mL)
Concentration after 24 hours
(CFU/mL)
(%)decline rate
(%)
녹농균Antibacterial test:
Pseudomonas aeruginosa
(0.05g/ml)Centella asiatica ethanol extract
(0.05g/ml)
폐렴간균Antibacterial test:
Klebsiella pneumoniae
(0.05g/ml)Centella asiatica ethanol extract
(0.05g/ml)
* CFU : Colony Forming Unit* 접종원 농도(CFU/mL) : 녹농균 : 4.3 x 106, 폐렴간균 : 8.3 x 106 * CFU: Colony Forming Unit* Inoculum concentration (CFU/mL): Pseudomonas aeruginosa: 4.3 x 10 6 , Klebsiella pneumoniae: 8.3 x 10 6
* 사용균주 : Pseudomonas aeruginosa ATCC 15442, Klebsiella pneumoniae ATCC 4352* Strains used: Pseudomonas aeruginosa ATCC 15442, Klebsiella pneumoniae ATCC 4352
* 반응시간 : 24시간* Response time: 24 hours
도 5는, 긴병꽃풀 에탄올 추출물(0.05 g/mL)의 녹농균 및 폐렴간균에 대한 항균시험 결과를 나타낸 사진이다.Figure 5 is a photograph showing the results of an antibacterial test against Pseudomonas aeruginosa and Klebsiella pneumoniae of the ethanol extract (0.05 g/mL) of Centella asiatica.
표 5 및 도 5를 참조하면, 긴병꽃풀 에탄올 추출물(0.05 g/mL)은 녹농균 및 폐렴간균에 대해 99.9%의 항균활성을 나타내는 것을 확인할 수 있다.Referring to Table 5 and Figure 5, it can be seen that the ethanol extract of Centella asiatica (0.05 g/mL) exhibits 99.9% antibacterial activity against Pseudomonas aeruginosa and Klebsiella pneumoniae.
2) 긴병꽃풀 에탄올 추출물(0.03 g/mL)에 대한 항균력 평가2) Evaluation of antibacterial activity of ethanol extract of Centella asiatica (0.03 g/mL)
(CFU/mL)Initial concentration
(CFU/mL)
(CFU/mL)
Concentration after 24 hours
(CFU/mL)
(%)decline rate
(%)
녹농균Antibacterial test:
Pseudomonas aeruginosa
(0.03g/ml)Centella asiatica ethanol extract
(0.03g/ml)
폐렴간균Antibacterial test:
Klebsiella pneumoniae
(0.03g/ml)Centella asiatica ethanol extract
(0.03g/ml)
* CFU : Colony Forming Unit* 접종원 농도(CFU/mL) : 녹농균 : 4.3 x 106, 폐렴간균 : 8.3 x 106 * CFU: Colony Forming Unit* Inoculum concentration (CFU/mL): Pseudomonas aeruginosa: 4.3 x 10 6 , Klebsiella pneumoniae: 8.3 x 10 6
* 사용균주 : Pseudomonas aeruginosa ATCC 15442, Klebsiella pneumoniae ATCC 4352* Strains used: Pseudomonas aeruginosa ATCC 15442, Klebsiella pneumoniae ATCC 4352
* 반응시간 : 24시간* Response time: 24 hours
도 6은, 긴병꽃풀 에탄올 추출물(0.03 g/mL)의 녹농균 및 폐렴간균에 대한 항균시험 결과를 나타낸 사진이다.Figure 6 is a photograph showing the results of an antibacterial test against Pseudomonas aeruginosa and Klebsiella pneumoniae of an ethanol extract (0.03 g/mL) of Centella asiatica.
표 6 및 도 6을 참조하면, 긴병꽃풀 에탄올 추출물(0.03 g/mL)은 녹농균 및 폐렴간균에 대해 99.9%의 항균활성을 나타내는 것을 확인할 수 있다.Referring to Table 6 and Figure 6, it can be seen that the ethanol extract of Centella asiatica (0.03 g/mL) exhibits 99.9% antibacterial activity against Pseudomonas aeruginosa and Klebsiella pneumoniae.
3) 긴병꽃풀 에탄올 추출물(0.02 g/mL)에 대한 항균력 평가3) Evaluation of antibacterial activity of ethanol extract of Centella asiatica (0.02 g/mL)
(CFU/mL)Initial concentration
(CFU/mL)
(CFU/mL)
Concentration after 24 hours
(CFU/mL)
(%)decline rate
(%)
녹농균Antibacterial test:
Pseudomonas aeruginosa
(0.02g/ml)Centella asiatica ethanol extract
(0.02g/ml)
폐렴간균Antibacterial test:
Klebsiella pneumoniae
(0.02g/ml)Centella asiatica ethanol extract
(0.02g/ml)
* CFU : Colony Forming Unit* 접종원 농도(CFU/mL) : 녹농균 : 4.3 x 106, 폐렴간균 : 8.3 x 106 * CFU: Colony Forming Unit* Inoculum concentration (CFU/mL): Pseudomonas aeruginosa: 4.3 x 10 6 , Klebsiella pneumoniae: 8.3 x 10 6
* 사용균주 : Pseudomonas aeruginosa ATCC 15442, Klebsiella pneumoniae ATCC 4352* Strains used: Pseudomonas aeruginosa ATCC 15442, Klebsiella pneumoniae ATCC 4352
* 반응시간 : 24시간* Response time: 24 hours
도 7은, 긴병꽃풀 에탄올 추출물(0.02 g/mL)의 녹농균 및 폐렴간균에 대한 항균시험 결과를 나타낸 사진이다.Figure 7 is a photograph showing the results of an antibacterial test against Pseudomonas aeruginosa and Klebsiella pneumoniae of the ethanol extract (0.02 g/mL) of Centella asiatica.
표 7 및 도 7을 참조하면, 긴병꽃풀 에탄올 추출물(0.02 g/mL)은 녹농균 및 폐렴간균에 대해 99.9%의 항균활성을 나타내는 것을 확인할 수 있다.Referring to Table 7 and Figure 7, it can be seen that the ethanol extract of Centella asiatica (0.02 g/mL) exhibits 99.9% antibacterial activity against Pseudomonas aeruginosa and Klebsiella pneumoniae.
4) 긴병꽃풀 에탄올 추출물(0.01 g/mL)에 대한 항균력 평가4) Evaluation of antibacterial activity of ethanol extract of Centella asiatica (0.01 g/mL)
(CFU/mL)Initial concentration
(CFU/mL)
(CFU/mL)
Concentration after 24 hours
(CFU/mL)
(%)decline rate
(%)
녹농균Antibacterial test:
Pseudomonas aeruginosa
(0.01g/ml)Centella asiatica ethanol extract
(0.01g/ml)
폐렴간균Antibacterial test:
Klebsiella pneumoniae
(0.01g/ml)Centella asiatica ethanol extract
(0.01g/ml)
* CFU : Colony Forming Unit* 접종원 농도(CFU/mL) : 녹농균 : 4.3 x 106, 폐렴간균 : 8.3 x 106 * CFU: Colony Forming Unit* Inoculum concentration (CFU/mL): Pseudomonas aeruginosa: 4.3 x 10 6 , Klebsiella pneumoniae: 8.3 x 10 6
* 사용균주 : Pseudomonas aeruginosa ATCC 15442, Klebsiella pneumoniae ATCC 4352* Strains used: Pseudomonas aeruginosa ATCC 15442, Klebsiella pneumoniae ATCC 4352
* 반응시간 : 24시간* Response time: 24 hours
도 8은, 긴병꽃풀 에탄올 추출물(0.01 g/mL)의 녹농균 및 폐렴간균에 대한 항균시험 결과를 나타낸 사진이다.Figure 8 is a photograph showing the results of an antibacterial test against Pseudomonas aeruginosa and Klebsiella pneumoniae of the ethanol extract (0.01 g/mL) of Centella asiatica.
표 8 및 도 8을 참조하면, 긴병꽃풀 에탄올 추출물(0.01 g/mL)은 녹농균 및 폐렴간균에 대해 99.9%의 항균활성을 나타내는 것을 확인할 수 있다.Referring to Table 8 and Figure 8, it can be seen that the ethanol extract of Centella asiatica (0.01 g/mL) exhibits 99.9% antibacterial activity against Pseudomonas aeruginosa and Klebsiella pneumoniae.
<실험예 3> 긴병꽃풀 에탄올 추출물의 피부자극 시험<Experimental Example 3> Skin irritation test of ethanol extract of Centipede chinensis
1) 실험 방법1) Experimental method
Rabbit에 대한 긴병꽃풀 70% (v/v) 에탄올 추출물의 피부자극을 평가하기 위하여, 0.5 mL의 시험 물질을 등 부위에 24 시간 첩포한 후(1일 1회), 72 시간 동안의 사망률, 일반 증상, 체중 변화 및 피부자극을 관찰 및 평가하였다.To evaluate the skin irritation of the 70% (v/v) ethanol extract of Centella asiatica to rabbits, 0.5 mL of the test substance was applied to the back area for 24 hours (once a day), and the mortality rate for 72 hours was determined. General symptoms, weight changes, and skin irritation were observed and evaluated.
a) 군 구성a) Military composition
(마리당)Administration site
(per animal)
시험부위 : 2Control area: 2
Test area: 2
b) 투여전 과정투여 약 24 시간 전에 등 부위를 제모 [약 (15 × 15) cm2]한 후, 투여 직전에 각각 [(2.5 × 2.5) cm2] 넓이의 투여부위 2 개소와 대조부위 2 개소로 구분 및 표시하였다. 실험동물 당 투여부위와 대조부위를 각각 찰과부위와 비찰과부위로 구분하였으며, 찰과부위는 표피만 손상되고 진피는 손상되지 않으며 피가 나지 않는 정도의 찰과상을 입혔다. b ) Pre-administration process Approximately 24 hours before administration, hair was removed on the back area [approximately (15 It is divided and marked by location. The administration site and control site per experimental animal were divided into an abrasion site and a non-abrasion site, respectively, and the abrasion site was an abrasion to the extent that only the epidermis was damaged, the dermis was not damaged, and there was no bleeding.
c) 시험물질 투여c) Test substance administration
0.5 mL의 시험물질을 gauze [(2.5 × 2.5) cm2]에 균일하게 도포하였다. 시험물질을 투여한 gauze를 상부 의 찰 과부위 1 개 소와 비찰과 부위 1 개소에 부 착하였으며, 하부의 대조부위에는 0.5 mL의 대조물질을 시험부위와 동일하게 부착하였다. 투여부위를 비자극성 테이프 (Tegaderm, 3M)와 탄력붕대 (Coban, 3M)를 이용하여 잘 고정시켰다. 첩포는 24 시간 후 제거하고 피부에 남아있는 시험물질은 세척물질 (멸균증류수)을 이용해 부드럽게 세정해주었다.0.5 mL of test substance was evenly applied to the gauze [(2.5 × 2.5) cm 2 ]. A gauze administered with the test substance was attached to one upper abrasion area and one non-abrasion area, and 0.5 mL of the control substance was attached to the lower control area in the same manner as the test area. The administration site was well fixed using non-irritating tape (Tegaderm, 3M) and an elastic bandage (Coban, 3M). The patch was removed after 24 hours, and the test substance remaining on the skin was gently washed using a cleaning agent (sterilized distilled water).
2) 실험 결과2) Experiment results
a) 사망률 및 일반 증상a) Mortality and general symptoms
모든 동물에서 시험물질 투여로 인한 사망동물은 관찰되지 않았으며, 일반증상 관찰결과, 시험물질 투여 후 24 시간에 전례에서 시험물질의 성상과 유사한 피부의 착색화 증상이 실험종료 시까지 관찰되었다.No dead animals were observed due to administration of the test substance in any of the animals, and as a result of observing general symptoms, symptoms of skin pigmentation similar to the properties of the test substance were observed 24 hours after administration of the test substance until the end of the experiment.
b) 체중변화b) Weight change
체중측정 결과, 시험물질 투여에 의한 체중변화는 관찰되지 않았다.As a result of weight measurement, no change in body weight was observed due to administration of the test substance.
c) 투여부위의 관찰 및 평가c) Observation and evaluation of the administration site
시험물질 투여부위를 관찰한 결과, 피부자극이 관찰되지 않았다. Draize의 평가방법에 따라 1차 피부자극지수 (P.I.I.)는 "0.0" 으로 산출되었다.As a result of observing the test substance administration site, no skin irritation was observed. According to Draize's evaluation method, the primary skin irritation index (P.I.I.) was calculated as "0.0".
상기 결과로부터 rabbit에 대한 피부자극시험에서 긴병꽃풀 70% 에탄올 추출물은 "비자극성(Non-irritant)" 물질로 판단되었다.From the above results, in the skin irritation test on rabbits, the 70% ethanol extract of Centella asiatica was judged to be a “non-irritant” substance.
<실험예 4> 긴병꽃풀 에탄올 추출물의 안점막자극 시험<Experimental Example 4> Eye mucous membrane irritation test of ethanol extract of Centella asiatica
1) 실험 방법1) Experimental method
Rabbit에 대한 긴병꽃풀 70% (v/v) 에탄올 추출물의 안점막자극을 평가하기 위하여 0.1 mL의 시험물질을 안구에 1 회 투여한 후 7 일 동안의 사망률, 일반증상, 체중변화 및 안점막자극을 관찰 및 평가하였다.To evaluate the ocular mucosal irritation of the 70% (v/v) ethanol extract of Centella asiatica in rabbits, mortality, general symptoms, weight changes, and ocular mucosa for 7 days were administered once to the eye with 0.1 mL of test substance. Stimuli were observed and evaluated.
a) 군 구성a) Military composition
b) 투여전 과정투여 약 24 시간 전에 양안의 각막, 홍채 및 결막 등의 이상유무를 휴대용 슬릿램프 (HEINE OPTOTECHNIK GmbH & Co. KG, 독일)를 이용하여 확인하였다.b) Pre-administration course Approximately 24 hours before administration, abnormalities in the cornea, iris, and conjunctiva of both eyes were checked using a portable slit lamp (HEINE OPTOTECHNIK GmbH & Co. KG, Germany).
c) 시험물질 투여c) Test substance administration
Rabbit을 보정한 후 9 마리의 우안에 시험물질 0.1 mL을 직접 투여하고, 좌안은 대조부위로 설정하였다. 투여 후 시험물질의 누출 방지를 위해 상하안검을 잡아 약 1 초간 폐안시켰다. 세척군 3 마리는 시험물질 투여 20 - 30 초 후, 양쪽 눈에 세척물질 (멸균생리식염수)를 사용하여 충분히 세척하였고, 나머지 6 마리는 세척하지 않았다.After calibrating the rabbit, 0.1 mL of test substance was directly administered to the right eyes of 9 rabbits, and the left eye was set as a control area. After administration, the upper and lower eyelids were held and the eyes closed for about 1 second to prevent leakage of the test substance. 3 animals in the washing group had both eyes thoroughly washed using a washing substance (sterilized saline solution) 20 to 30 seconds after administration of the test substance, and the remaining 6 animals were not washed.
2) 실험 결과2) Experiment results
a) 사망률 및 일반증상a) Mortality rate and general symptoms
실험기간 중 시험물질 투여로 인한 사망동물은 관찰되지 않았고, 모든 동물에서 특이할 만한 이상증상은 관찰되지 않았다.No dead animals were observed due to test substance administration during the experiment period, and no unusual abnormal symptoms were observed in any animal.
b) 체중변화b) Weight change
체중측정 결과, 모든 동물에서 정상적인 체중증가가 관찰되었다.As a result of weight measurement, normal weight gain was observed in all animals.
c) 투여부위의 관찰 및 평가c) Observation and evaluation of the administration site
시험물질 투여부위를 관찰한 결과, 비세척군 및 세척군 모두에서 안점막자극이 관찰되지 않았다. Draize의 평가방법에 따른 급성안점막자극지수 (I.A.O.I.)는 비세척군 및 세척군에서 모두 "0.0" 으로 산출되었다.As a result of observing the test substance administration site, no eye mucosa irritation was observed in both the non-washed and washed groups. The acute ocular mucosal irritation index (I.A.O.I.) according to Draize's evaluation method was calculated as "0.0" in both the non-washed and washed groups.
상기 결과로부터 rabbit에 대한 안점막자극시험에서 긴병꽃풀 70% 에탄올 추출물은 "무자극물(Nonirritating)" 물질로 판단되었다.From the above results, the 70% ethanol extract of Centella asiatica was judged to be a "nonirritating" substance in the eye mucosa irritation test on rabbits.
<실험예 5> 긴병꽃풀 에탄올 추출물의 단회경구투여 독성시험<Experimental Example 5> Single oral administration toxicity test of ethanol extract of Centella asiatica
1) 실험 방법1) Experimental method
긴병꽃풀 70% (v/v) 에탄올 추출물에 대한 단회경구투여독성시험을 실시하기 위해 SD계 rat를 사용하여 식품의약품안전처 고시“의약품등의 독성시험기준”에 따라 2000 mg/kg B.W. 용량을 최고용량으로 설정하고 공비를 2 로 두어 1000 및 500 mg/kg B.W. 으로 중용량 및 저용량을 설정하여 암·수 각각에 단회경구투여 하였다.To conduct a single oral dose toxicity test on the 70% (v/v) ethanol extract of Centella asiatica, SD rats were used and 2000 mg/kg B.W. Set the dose to the highest dose and set the common ratio to 2 to obtain 1000 and 500 mg/kg B.W. Medium and low doses were set and administered as a single oral dose to males and females, respectively.
시험물질 투여 후 14 일간 사망률, 일반증상, 체중변화를 관찰하였으며, 생존동물은 부검하여 육안적으로 장기의 이상 유무를 검사하였다.Mortality, general symptoms, and weight changes were observed for 14 days after test substance administration, and surviving animals were autopsied and visually inspected for organ abnormalities.
a) 군 구성a) Military composition
(mg/kg B.W.)dosage
(mg/kg BW)
(mL/kg B.W.)Dosage amount
(mL/kg BW)
※ G1 : 부형제 대조군, G2 - G4 : 시험물질 투여군, B.W. : Body weightb) 시험물질의 투여 경로 및 방법※ G1: Excipient control group, G2 - G4: Test substance administration group, B.W. : Body weightb) Administration route and method of test substance
약 18 시간 절식시킨 실험동물에 시험물질 조제물을 경구투여용 sonde를 장착한 주사기를 이용하여 위내에 1 회 강제 투여하였다.The test substance preparation was forcibly administered once into the stomach to experimental animals that were fasted for about 18 hours using a syringe equipped with a sonde for oral administration.
c) 관찰항목c) Observation items
일반증상은 모든 동물에 대하여 1일 1회 관찰 하였으며, 시험물질 투여 후 14 일간 관찰하였다. 단, 투여 당일에는 투여 후 0.5, 1, 2, 3 및 4 시간에 관찰하였다.General symptoms were observed once a day for all animals, and were observed for 14 days after administration of the test substance. However, on the day of administration, observations were made at 0.5, 1, 2, 3, and 4 hours after administration.
체중은 도입 시, 군 분리 시, 시험물질투여 개시 직전, 투여 개시 후 7 일 및 14 일째에 측정하였다.Body weight was measured at introduction, group separation, immediately before the start of test substance administration, and on the 7th and 14th days after the start of administration.
부검은 투여 후 14 일째 모든 생존동물의 외관 검사를 실시한 후, Isoflurane (아이프란액, 하나제약) 마취하에 방혈치사 하여 육안으로 장기를 검사하였다.Autopsies were conducted on the 14th day after administration, and all surviving animals were examined externally. Then, they were killed by exsanguination under isoflurane (Ifrane, Hana Pharmaceutical) anesthesia and their organs were inspected with the naked eye.
2) 실험 결과2) Experiment results
실험기간 중 시험물질 투여에 의한 사망동물은 관찰되지 않았고, 일반증상 관찰결과, 시험물질 투여에 의한 이상소견은 관찰되지 않았다.No dead animals were observed due to test substance administration during the experiment period, and as a result of observing general symptoms, no abnormal findings were observed due to test substance administration.
체중측정 결과, 모든 시험물질 투여군에서 부형제 대조군과 비교하여 통계학적으로 유의 적인 차이는 관찰되지 않았고, 부검소견 결과, 모든 투여군에서 시험물질 투여에 의한 이상소견은 관찰되지 않았다.As a result of weight measurement, no statistically significant difference was observed in all test substance administration groups compared to the excipient control group, and as a result of autopsy findings, no abnormal findings due to test substance administration were observed in all treatment groups.
상기 결과로부터 rat에 긴병꽃풀 70% 에탄올 추출물을 단회경구투여 시 시험물질 투여와 관련된 독성학적 소견이 인정되지 않았으므로 개략의 치사량은 암·수 모두 2000 mg/kg B.W.으로 판단되었다.From the above results, since no toxicological findings related to the administration of the test substance were recognized when the 70% ethanol extract of Centella asiatica was administered once orally to rats, the approximate lethal dose was determined to be 2000 mg/kg B.W. for both males and females.
<실험예 6> 긴병꽃풀 에탄올 추출물의 단회경피투여 독성시험<Experimental Example 6> Single transdermal administration toxicity test of ethanol extract of Centella asiatica
1) 실험 방법1) Experimental method
긴병꽃풀 70% (v/v) 에탄올 추출물에 대한 단회경피투여독성시험을 실시하기 위해 SD계 rat를 사용하여 식품의약품안전처 고시“의약품등의 독성시험기준”에 따라 2000 mg/kg B.W. 용량을 최고용량으로 설정하고 공비를 2 로 두어 1000 및 500 mg/kg B.W. 으로 중용량 및 저용량을 설정하여 암·수 각각에 단회경피투여 하였다.In order to conduct a single transdermal dose toxicity test on the 70% (v/v) ethanol extract of Centella asiatica, SD rats were used and 2000 mg/kg B.W. Set the dose to the highest dose and set the common ratio to 2 to obtain 1000 and 500 mg/kg B.W. Medium and low doses were set and administered once transdermally to males and females, respectively.
시험물질 투여 후 14 일간 사망률, 일반증상, 체중변화를 관찰하였으며, 생존동물은 부검하여 육안적으로 장기의 이상 유무를 검사하였다.Mortality, general symptoms, and weight changes were observed for 14 days after test substance administration, and surviving animals were autopsied and visually inspected for organ abnormalities.
a) 군 구성a) Military composition
(mg/kg B.W.)dosage
(mg/kg BW)
(mL/kg B.W.)Dosage amount
(mL/kg BW)
※ G1 : 부형제 대조군, G2 - G4 : 시험물질 투여군, B.W. : Body weightb) 시험물질의 투여 경로 및 방법※ G1: Excipient control group, G2 - G4: Test substance administration group, B.W. : Body weightb) Administration route and method of test substance
투여 전날 실험동물의 등 부위를 가급적 피부가 손상 되지 않도록 넓게 제모하였다. 시험물질의 도포면적은 체표면적의 10 % 정도 [rat : (4 × 4) cm2]로 하였다. 시험물질 조제물을 gauze에 적용 후 제모한 부위에 부착하고, 시험물질의 유실을 막기 위해 비자극성 테이프 (Tegaderm, 3M)와 탄력붕대 (Coban, 3M)로 24 시간 고정하였다. 노출 종료 후 도포물을 제거하고 피부에 남아있는 시험물질 조제물을 멸균증류수 (주사용수)로 잘 닦아주었다.The day before administration, the back area of the experimental animal was removed as widely as possible to avoid damaging the skin. The application area of the test substance was approximately 10% of the body surface area [rat: (4 × 4) cm2]. The test substance preparation was applied to the gauze and attached to the hair-removed area, and fixed for 24 hours with non-irritating tape (Tegaderm, 3M) and an elastic bandage (Coban, 3M) to prevent loss of the test substance. After exposure was completed, the application was removed, and the test substance preparation remaining on the skin was thoroughly wiped off with sterile distilled water (water for injection).
c) 관찰항목c) Observation items
일반증상은, 모든 동물에 대하여 1일 1회 관찰하였으며, 시험물질 투여 후 14 일간 관찰하였다. 단, 투여 당일에는 투여 후 0.5, 1, 2, 3 및 4 시간에 관찰하였다.General symptoms were observed for all animals once a day for 14 days after administration of the test substance. However, on the day of administration, observations were made at 0.5, 1, 2, 3, and 4 hours after administration.
체중은 도입 시, 군 분리 시, 시험물질투여 개시 직전, 투여 개시 후 7 일 및 14 일째에 측정하였다.Body weight was measured at introduction, group separation, immediately before the start of test substance administration, and on the 7th and 14th days after the start of administration.
부검은 투여 후 14 일째 모든 생존동물의 외관 검사를 실시한 후, Isoflurane (아이프란액, 하나제약) 마취하에 방혈치사 하여 육안으로 장기를 검사하였다.Autopsies were conducted on the 14th day after administration, and all surviving animals were examined externally. Then, they were killed by exsanguination under isoflurane (Ifrane, Hana Pharmaceutical) anesthesia and their organs were inspected with the naked eye.
2) 실험 결과2) Experiment results
실험기간 중 시험물질 투여에 의한 사망동물은 관찰되지 않았고, 일반증상 관찰결과, 암컷 1000 m g/kg B.W . 시험물 질 투여군 및 암 ·수컷 2000 m g/kg B.W . 시험물질 투여군에서 관찰된 피부의 착색화가 관찰되었다.During the experiment period, no dead animals were observed due to test substance administration, and as a result of general symptom observation, the female was 1000 m g/kg B.W. Test substance administration group and male and female 2000 m g/kg B.W. Pigmentation of the skin was observed in the test substance administered group.
체중측정 결과, 모든 시험물질 투여군에서 부형제 대조군과 비교하여 통계학적으로 유의적인 차이는 관찰되지 않았고, 부검소견 결과, 모든 투여군에서 시험물질 투여에 의한 이상소견은 관찰되지 않았다.As a result of body weight measurement, no statistically significant difference was observed in all test substance administration groups compared to the excipient control group, and as a result of autopsy findings, no abnormal findings due to test substance administration were observed in all administration groups.
상기 결과로부터 rat에 긴병꽃풀 70% 에탄올 추출물을 단회경피투여 시 시험물질 투여와 관련된 독성학적 소견이 인정되지 않았으므로 개략의 치사량은 암·수 모두 2000 mg/kg B.W. 으로 판단된다.From the above results, no toxicological findings related to test substance administration were recognized when a single transdermal administration of 70% ethanol extract of Centella asiatica to rats, so the approximate lethal dose was 2000 mg/kg B.W. for both males and females. It is judged as
<실험예 7> 긴병꽃풀 지상부 추출물의 폴리페놀 함량 측정<Experimental Example 7> Measurement of polyphenol content of aerial part extract of Longasia chinensis
상기 제조된 긴병꽃풀 지상부 추출물의 총 폴리페놀 함량은 Folin-Denis 방법을 변형하여 측정하였다. 농축하지 않은 추출물 10 μL를 취하여 10% sodium carbonate (Na2CO3) 200 μL와 1N Folin-ciocalteu reagent 10 μL와 혼합하여 상온에서 30분 반응 후 750 nm에서의 흡광도를 측정하였다. The total polyphenol content of the above-prepared extract of the above-mentioned aerial parts of Centella asiatica was measured by modifying the Folin-Denis method. 10 μL of the unconcentrated extract was taken and mixed with 200 μL of 10% sodium carbonate (Na 2 CO 3 ) and 10 μL of 1N Folin-ciocalteu reagent. After reaction for 30 minutes at room temperature, the absorbance was measured at 750 nm.
폴리페놀 함량은 galic acid를 표준물질로 한 표준곡선을 통해 계산하였다.Polyphenol content was calculated through a standard curve using galic acid as a standard material.
본 발명의 일 실시형태에 따른 긴병꽃풀 지상부 추출물(긴병꽃풀 70% (v/v) 에탄올 추출물)의 총 폴리페놀 함량을 측정한 결과, 추출물의 총 폴리페놀 함량은 88.44 mg/g GAE로 나타났다.As a result of measuring the total polyphenol content of the aerial part of the extract of Centella asiatica (70% (v/v) ethanol extract of Centella asiatica) according to one embodiment of the present invention, the total polyphenol content of the extract was 88.44 mg/g GAE. appear.
이상과 같이 실시예들이 비록 한정된 도면에 의해 설명되었으나, 해당 기술분야에서 통상의 지식을 가진 자라면 상기를 기초로 다양한 기술적 수정 및 변형을 적용할 수 있다. 예를 들어, 설명된 기술들이 설명된 방법과 다른 순서로 수행되거나, 및/또는 설명된 시스템, 구조, 장치, 회로 등의 구성요소들이 설명된 방법과 다른 형태로 결합 또는 조합되거나, 다른 구성요소 또는 균등물에 의하여 대치되거나 치환되더라도 적절한 결과가 달성될 수 있다.Although the embodiments have been described with limited drawings as described above, those skilled in the art can apply various technical modifications and variations based on the above. For example, the described techniques are performed in a different order than the described method, and/or components of the described system, structure, device, circuit, etc. are combined or combined in a different form than the described method, or other components are used. Alternatively, appropriate results may be achieved even if substituted or substituted by an equivalent.
그러므로, 다른 구현들, 다른 실시예들 및 특허청구범위와 균등한 것들도 후술하는 청구범위의 범위에 속한다.Therefore, other implementations, other embodiments, and equivalents of the claims also fall within the scope of the following claims.
Claims (9)
상기 긴병꽃풀 지상부 추출물은,
폴리페놀 5 중량% 내지 10 중량%를 포함하고,
폐렴간균(Klebsiella pneumoniae), 녹농균(Pseudomonas aeruginosa) 및 대장균(Escherichia coli) 중 적어도 하나 이상의 균주에 대하여 항균 활성을 갖고,
상기 긴병꽃풀 지상부 추출물의 농도는,
0.01 g/ml 내지 0.05 g/ml이고,
상기 긴병꽃풀 지상부 추출물의 추출은,
20 ℃ 내지 30 ℃의 온도 및 0.1 MPa 내지 0.2 Mpa 압력 조건에서 24 시간 내지 48 시간 동안 수행되는 것인,
항균제 조성물.
Contains the above-ground extract of Glechoma grandis (A. Gray) Kuprian as an active ingredient,
The above-mentioned extract of the above-mentioned part of Longasia chinensis,
Contains 5% to 10% by weight of polyphenol,
Has antibacterial activity against at least one strain of Klebsiella pneumoniae , Pseudomonas aeruginosa , and Escherichia coli ,
The concentration of the above-mentioned extract of the above-mentioned long-distance plant is,
0.01 g/ml to 0.05 g/ml,
Extraction of the above-mentioned extract of the above-mentioned part of Longhorn Flower,
It is carried out for 24 hours to 48 hours at a temperature of 20 ℃ to 30 ℃ and a pressure of 0.1 MPa to 0.2 Mpa,
Antibacterial composition.
상기 긴병꽃풀 지상부 추출물은,
물, 탄소수 1 내지 4의 저급 알코올, 아세톤, 에틸아세테이트, 클로로포름, 1,3-부틸렌글리콜, n-헥산, 디에틸에테르, 부틸아세테이트 및 디클로로메탄으로 이루어진 군에서 선택되는 하나 이상을 추출 용매로 하여 추출된 것인,
항균제 조성물.
According to paragraph 1,
The above-mentioned extract of the above-mentioned part of Longasia chinensis,
At least one selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms, acetone, ethyl acetate, chloroform, 1,3-butylene glycol, n-hexane, diethyl ether, butyl acetate, and dichloromethane as the extraction solvent. Extracted from this,
Antibacterial composition.
상기 저급 알코올은, 에탄올인 것인,
항균제 조성물.
According to paragraph 4,
The lower alcohol is ethanol,
Antibacterial composition.
상기 항균제 조성물은,
용액, 현탁액, 유탁액, 페이스트, 겔, 크림, 로션, 파우더, 비누, 계면활성제 함유 클렌징, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션 및 스프레이로 이루어진 군에서 선택되는 하나 이상의 제형을 포함하는 것인,
항균제 조성물.
According to paragraph 1,
The antibacterial composition is,
Containing one or more formulations selected from the group consisting of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, cleansers containing surfactants, oils, powder foundations, emulsion foundations, wax foundations and sprays. person,
Antibacterial composition.
항균용 피부 외용제.Containing the antibacterial composition of claim 1,
Antibacterial external skin preparation.
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| Title |
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| https://14128625.tistory.com/272, 일상을프레임에가두다, 긴병꽃풀, 2017.01.27. |
| 임수빈 외 6명. 긴병꽃풀의 생리활성과 이를 첨가한 양갱의 품질특성. 한국식품저장유통학회, 2017 |
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