KR20130070683A - Composition containing extract of bower actinidia which has antioxidant activity - Google Patents
Composition containing extract of bower actinidia which has antioxidant activity Download PDFInfo
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- KR20130070683A KR20130070683A KR1020110137318A KR20110137318A KR20130070683A KR 20130070683 A KR20130070683 A KR 20130070683A KR 1020110137318 A KR1020110137318 A KR 1020110137318A KR 20110137318 A KR20110137318 A KR 20110137318A KR 20130070683 A KR20130070683 A KR 20130070683A
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- Prior art keywords
- extract
- antioxidant
- fraction
- activity
- present
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Abstract
Description
본 발명은 다래나무 과에 속하는 덩굴성 낙엽만목인 다래나무(Actinidia arguta Planchon)의 잎 또는 줄기로부터 추출한 에탄올 추출물 또는 상기 다래나무 에탄올 추출물의 항산화 활성 분획 물을 유효 성분으로 함유하는 항산화용 약제학적 조성물, 항산화용 기능성 식품 조성물 및 항산화용 화장품 조성물에 관한 것이다. 더욱 상세하게는 다래나무 분말에 70% 에탄올을 가하여 24시간동안 실온에서 진탕하여 2회 반복 추출한 후 여지로 여과한 뒤 농축기로 40℃에서 감압농축 및 동결건조 하여 70% 에탄올 추출물을 얻은 후, 물에 용해시켜 분액깔때기에 넣은 뒤 n-hexane, Chloroform, n-butanol 및 Aqueous 용매의 순서로 극성의 차이를 이용하여 분획 물을 얻었으며 분리된 각각의 용매 분획 물은 감압 농축하여 용매를 제거 한 후 -70℃에서 동결 건조하여 각 분획의 수율을 구하였다. 이후 최종적으로 분획 물을 대상으로 총 폴리페놀, 플라보노이드 함량과 항산화 활성(DPPH 라디칼 소거 능, ABTS 라디칼 소거 능, SOD 유사활성)을 측정하였고, 합성 항산화제인 BHT와 천연 항산화제인 ascorbic acid 및 α-tocopherol과 비교함으로써 다래 분획 추출물의 항산화활성을 규명하였다.The present invention is a vine of deciduous deciduous tree belonging to the genus Perilla ( Acinidia arguta The present invention relates to a pharmaceutical composition for antioxidant, a functional food composition for antioxidant, and a cosmetic composition for antioxidant, which contain an ethanol extract extracted from the leaves or stems of Planchon ) or an antioxidant active fraction of the ethanol extract of Mt. More specifically, 70% ethanol was added to the powder of the taraxacum, shaken at room temperature for 24 hours, extracted twice, filtered and filtered. After dissolving in a separatory funnel, fractions were obtained using polarity difference in the order of n-hexane, Chloroform, n-butanol, and Aqueous solvent. Each solvent fraction was concentrated under reduced pressure to remove solvent. Lyophilization at -70 ° C yielded the yield of each fraction. Finally, the total polyphenol, flavonoid content and antioxidant activity (DPPH radical scavenging ability, ABTS radical scavenging ability, SOD-like activity) were measured in fractions, and the synthetic antioxidants BHT and natural antioxidants ascorbic acid and α-tocopherol were measured. The antioxidant activity of the extract of Darae fraction was investigated by comparing with.
자유 라디칼은 쌍을 이루지 않은 전자를 갖는 원자, 이온, 또는 분자로서, 자유 라디칼, 특히 산소 분자로부터 유도된 자유 라디칼은 여러 가지 생물학적 현상에 있어서 기본적인 기능을 한다. 다양한 형태의 산소와 자유 라디칼의 농도가 과다할 경우, 생체 내에 막 지질의 과산화, 핵산 염기의 히드록실 화 및 설프히드릴기와 기타 단백질 내의 라디칼 활성 부분의 산화와 같은 심각한 역효과를 미칠 수 있으며, 이들이 억제되지 않을 경우, 변이와 세포 사멸을 초래할 수 있다.Free radicals are atoms, ions, or molecules that have unpaired electrons. Free radicals, especially free radicals derived from oxygen molecules, play a fundamental role in many biological phenomena. Excessive concentrations of various types of oxygen and free radicals can have severe adverse effects such as peroxidation of membrane lipids in vivo, hydroxylation of nucleic acid bases and oxidation of radical active moieties in sulfhydryl groups and other proteins, If not suppressed, it can lead to mutations and cell death.
많은 자유 라디칼 반응은 생체분자에 산화 손상을 일으키며, 단백질을 가교시키고, DNA를 변이시키며, 지질을 과 산화시킨다. 자유 라디칼이 일단 형성되면, 이들은 상호작용에 의해 다른 자유 라디칼 및 비 라디칼 산화제 등을 생성한다. 또한, 자유 라디칼 반응의 생성물 중 일부가 손상되면 잠재적으로 손상을 일으키는 화학종을 생성할 수 있다. 산소 자유 라디칼은 또한 단백질의 산화 변성을 일으킨다. 또한, 자유 라디칼의 산화 손상은 알츠하이머병, 류마티스 관절염, 암 및 죽상동맥경화증과 같은 노화 및 퇴행성 질환에서 주원인으로 작용한다고 보고되고 있다.Many free radical reactions cause oxidative damage to biomolecules, crosslinking proteins, mutating DNA, and oxidizing lipids. Once free radicals are formed, they interact with other free radicals and non-radical oxidizing agents. In addition, if some of the products of the free radical reaction are damaged, they can produce chemical species that can potentially cause damage. Oxygen-free radicals also cause oxidative modification of the protein. In addition, oxidative damage of free radicals has been reported to play a major role in aging and degenerative diseases such as Alzheimer's disease, rheumatoid arthritis, cancer and atherosclerosis.
이에, 항산화 작용을 하는 여러 물질에 대한 요구 및 항산화 관련 화장품, 항산화 관련 기능성 식품 및 항산화 관련 약제 조성물에 대한 수요가 증가하고 있고, 그와 관련한 연구가 활발히 진행되고 있으며, 신규 항산화 물질을 발견하기 위한 노력이 계속되고 있다.Accordingly, there is a growing demand for various antioxidant substances, antioxidant-related cosmetics, antioxidant-related functional foods and antioxidant-related pharmaceutical compositions, and researches related thereto have been actively conducted. Efforts are continuing.
다래나무(Bower actinidia)는 다래나무과에 속하는 덩굴성 낙엽만목으로 학명은 Actinidia arguta PLANCH.이다. 우리나라·중국·일본에 자생하며 깊은 산의 숲속, 토양이 깊고 부식질이 많은 곳에서 잘 자란다. 줄기의 길이는 7m에 달하고 어린 가지에는 잔털이 있으며, 잎은 어긋나고 넓은 난형 또는 타원형이다. 꽃은 2가화(二家花)로서 5월에 피는데 백색으로 3∼10개가 달린다. 열매는 난상(卵狀) 원형으로 10월에 황록색으로 익는데 맛이 좋다.Bower actinidia is a deciduous deciduous tree belonging to the family Tetaceae, and its scientific name is Actinidia. arguta PLANCH. to be. It is native to Korea, China, and Japan, and grows well in deep mountain forests and in places with deep soils and humus. Stem length is 7m, young branches have fine hairs, leaves are alternate, broad ovate or oval. Flowers are biphasic (二 家 花), bloom in May, white to 3-10. Fruits are egg-shaped round, ripened in yellow-green in October, and have good taste.
다래 열매는 단맛이 있으므로 생식되고 있으며, 뜨거운 물을 끼얹어 건조시킨 것을 달여 마시면 신경통에도 좋다고 한다. 특히, 열매에 충영(蟲?: 혹처럼 생긴 식물체)이 생긴 것은 목천료(木天蓼)라 하여 귀중한 한약재로 여긴다.Tadasu fruit is sweet because of its sweetness, and it is said to be good for neuralgia when it is dried and drunk with hot water. In particular, the fruit (chung ?: lump-like plant) in the fruit is called Mokcheonryu (木 天 蓼) is considered a valuable herbal medicine.
다래주도 몸을 따뜻하게 해주며, 이뇨·강심·강장 등에 효능이 있다고 한다. 다래나무의 껍질과 줄기는 노끈으로 대용되기도 한다.Dadaju warms the body and is said to be effective in diuresis, cardio, and tonic. The bark and stems of the stalks are sometimes used as strings.
이와 같이 다래는 우리 민족이 즐겨 먹던 산과(山果)로 문학작품에서는 머루와 짝이 되어 나타난다. 특히 고려가요인 「청산별곡」에서 보이는 다래·머루는 평원의 생활에서 주식으로 삼는 쌀·보리와 상대적인 개념으로 이해된다.Like this, Darae is a mountain fruit that our people enjoyed. In particular, Darae and Maru, which are seen in the Goryeo Song, “Clearing Stars,” are understood as a concept relative to rice and barley, which are used as staples in plain life.
또한 다래나무의 잎과 줄기의 경우 생체의 노화 과정과 관련성이 있는 유리기 및 만성질환의 피해를 막아주는 산화 방지 역할을 하는 페놀과 항균, 항암, 항바이러스, 항알레르기 및 항염증 활성을 나타내는 플라보노이드를 다량 함유하고 있어 상기와 같은 항산화 물질에 대한 요구 및 수요를 충족시킬 수 있으며, 천연물이기에 비교적 안전성이 높고 부작용이 적다는 장점이 있다. 이에 상기 요구 및 수요에 대한 적절한 충족 및 공급을 위해, 본 연구에서는 우리나라, 중국 및 일본 등지에 자생하는 다래나무의 잎 또는 줄기로부터 항산화 활성 분획물을 수득하여 항산화 활성을 규명하고 항산화 활성 분획물을 유효 성분으로 함유하는 항산화용 약제학적 조성물, 항산화용 기능성 식품 조성물 및 항산화용 화장품 조성물을 완성하였다.In addition, the leaves and stems of the taraxacum are phenols that act as antioxidants to prevent the damage of free radicals and chronic diseases related to the aging process of the living body, and flavonoids that exhibit antibacterial, anticancer, antiviral, antiallergic and anti-inflammatory activities. Because it contains a large amount can meet the demands and demands for the above-mentioned antioxidants, because it is a natural product has the advantage of relatively high safety and fewer side effects. In order to satisfy and supply the demands and demands, the present study obtained antioxidant activity fractions from the leaves or stems of wild rapeseed trees in Korea, China, Japan, etc. A pharmaceutical composition for antioxidant, a functional food composition for antioxidant, and a cosmetic composition for antioxidant were completed.
본 명세서 전체에 걸쳐 다수의 논문 및 특허문헌이 참조되고 그 인용이 표시되어 있다. 인용된 논문 및 특허문헌의 개시 내용은 그 전체로서 본 명세서에 참조로 삽입되어 본 발명이 속하는 기술 분야의 수준 및 본 발명의 내용이 보다 명확하게 설명된다.Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.
본 발명자들은 상기와 같은 항산화 작용물질 요구를 충족시키기 위한 용도의 천연물을 찾기 위하여 연구 노력하였다. 그 결과, 다래나무의 잎 또는 줄기로부터 수득한 항산화 활성 분획물이 페놀 및 플라보노이드를 다량 함유하여 항산화 활성이 우수하고 생체 내 부작용이 적다는 것을 실험적으로 확인하여 본 발명을 완성하였다.The present inventors have made an effort to find natural products for use to satisfy the above antioxidant demands. As a result, the present invention was completed by experimentally confirming that the antioxidant active fractions obtained from the leaves or stems of the larvae contain a large amount of phenol and flavonoids, which have excellent antioxidant activity and fewer side effects in vivo.
따라서, 본 발명의 목적은 다래나무의 잎 또는 줄기로부터 추출한 에탄올 용매 추출물 또는 상기 에탄올 추출물로부터 수득한 항산화 활성 분획물을 제공하는 것에 있다.Accordingly, it is an object of the present invention to provide an ethanol solvent extract extracted from the leaves or stems of a stalk tree or an antioxidant active fraction obtained from the ethanol extract.
본 발명의 다른 목적은 상기의 에탄올 추출물 및 항산화 활성 분획물을 유효 성분으로 함유하는 항산화용 약제학적 조성물, 항산화용 기능성 식품 조성물 및 항산화용 화장품 조성물을 제공하는 것에 있다.Another object of the present invention is to provide a pharmaceutical composition for antioxidant, functional food composition for antioxidant and cosmetic composition for antioxidant containing the ethanol extract and the antioxidant active fraction as an active ingredient.
본 발명의 목적 및 장점은 하기의 발명의 상세한 성명, 청구의 범위 및 도면에 의해 보다 명확하게 된다.The objects and advantages of the invention will become apparent from the following detailed description, claims and drawings.
본 발명의 일 양태에 따르면, 항산화 활성이 있는 다래나무(Actinidia arguta Planchon) 에탄올 추출물을 제공한다.According to one aspect of the present invention, there is provided an ethanol extract of Actinidia arguta Planchon with antioxidant activity.
본 발명의 명세서에서 용어 ‘다래나무(Bower Actinidia)’는 쌍떡잎식물 측막태좌목 다래나무과의 낙엽 덩굴나무를 의미한다.The term 'Bower Actinidia' in the context of the present invention refers to the deciduous vines of the dicotyledonous plant, the alveolar cucurbita.
본 발명의 명세서에서 용어 ‘항산화(抗酸化)’는 생체분자에 산화 손상을 일으키며, 단백질을 가교시키고, DNA를 변이시키며, 지질을 과 산화시킬 수 있는 자유 라디칼 및 활성 산소를 억제하는 것을 의미한다.The term 'antioxidant' in the context of the present invention means inhibiting free radicals and free radicals that can cause oxidative damage to biomolecules, crosslink proteins, mutate DNA, and overoxidize lipids. .
본 발명의 명세서에서 용어 ‘활성(活性)’은 자유 라디칼 및 활성 산소를 억제하는 항산화 효과의 활동이 활발하여 지거나 반응 속도가 빨라지는 성질을 의미한다.In the present specification, the term 'active' refers to a property in which an activity of an antioxidant effect of inhibiting free radicals and active oxygen becomes active or a reaction rate is accelerated.
본 발명의 명세서에서 용어 ‘에탄올(Ethanol)’은 사슬모양의 탄화수소 중 탄소의 수가 2개인 에테인에서 수소 원자 하나가 하이드록시기로 치환된 알코올로, 휘발성과 가연성을 가진 무색 액체를 의미한다.In the present specification, the term 'ethanol' refers to a colorless liquid having volatile and flammable alcohols in which one hydrogen atom is substituted with a hydroxyl group in ethane having two carbon atoms in the chain-shaped hydrocarbon.
본 발명의 명세서에서 용어 ‘추출물(抽出物)’은 다래나무의 잎 또는 줄기의 분말을 용매에 녹여 분리한 물질을 의미한다.In the specification of the present invention, the term 'extract (抽出 物)' refers to a material obtained by dissolving the powder of the leaves or stems of a lychee tree in a solvent.
본 발명자들은 통상의 항산화 관련 약제 조성물, 화장품 조성물 및 기능성 식품 조성물들보다 항산화 효능이 개선된 항산화 물질을 제조하기 위하여 예의 연구 노력하였다. 그 결과, 상술한 장점을 가지는 다래나무 에탄올 추출물을 수득하였다. 다래나무의 잎 또는 줄기의 분말에 70% 에탄올을 가하여 24시간동안 실온에서 진탕하여 2 내지 3회 반복 추출한 후 여지로 여과한 뒤 농축기로 40℃에서 감압농축 및 동결건조 하여 70% 에탄올 추출물을 얻은 후, 이 에탄올 용매 추출물을 물에 용해시켜 분액깔때기에 넣은 뒤 앤-헥산(n-hexane), 클로로포름(Chloroform), 앤-부탄올(n-butanol) 및 에이퀴어스(Aqueous)를 포함하는 군으로부터 선택된 용매를 사용하여 항산화 활성 분획 물을 얻었으며 분리된 각각의 용매 분획 물은 감압 농축하여 용매를 제거 한 후 -70℃에서 동결 건조하여 각 분획의 수율을 구하였다. 이후 최종적으로 분획 물을 대상으로 총 폴리페놀, 플라보노이드 함량과 항산화 활성(DPPH 라디칼 소거 능, ABTS 라디칼 소거 능, SOD 유사활성)을 측정하였고, 합성 항산화제인 BHT와 천연 항산화제인 ascorbic acid 및 α-tocopherol과 비교함으로써 다래 분획 추출물의 항산화활성을 확인하였으며, 상기 추출물 및 분획 물을 유효성분으로 함유하는 항산화 관련 약제 조성물, 화장품 조성물 및 기능성 식품 조성물의 경우, 항산화 효능이 뛰어나며 생체 내에 어떤 부작용도 일으키지 않는다는 것을 확인하였다.The present inventors have made intensive studies to prepare antioxidant substances having improved antioxidant efficacy over conventional antioxidant related pharmaceutical compositions, cosmetic compositions, and functional food compositions. As a result, the ethanol extract of the cedar tree having the above-mentioned advantages was obtained. 70% ethanol was added to the leaves or stem powders of the trees, shaken at room temperature for 24 hours, and extracted two to three times. After filtration, the mixture was concentrated under reduced pressure and freeze-dried at 40 ° C. with a concentrator to obtain 70% ethanol extract. Then, the ethanol solvent extract was dissolved in water and placed in a separatory funnel, and then from the group containing n-hexane, chloroform, n-butanol and Aqueous. Antioxidant active fractions were obtained using the selected solvent, and each of the separated solvent fractions was concentrated under reduced pressure to remove the solvent, and then freeze-dried at -70 ° C to obtain the yield of each fraction. Finally, the total polyphenol, flavonoid content and antioxidant activity (DPPH radical scavenging ability, ABTS radical scavenging ability, SOD-like activity) were measured in fractions, and the synthetic antioxidants BHT and natural antioxidants ascorbic acid and α-tocopherol were measured. The antioxidative activity of the extract of Darae fraction was confirmed by comparison with that of the antioxidant-related pharmaceutical composition, the cosmetic composition and the functional food composition containing the extract and the fraction as an active ingredient, showing that the antioxidant activity is excellent and does not cause any side effects in vivo. Confirmed.
본 발명의 바람직한 양태에 따르면, 상기 다래나무는 다래나무의 잎 또는 줄기를 의미한다.According to a preferred embodiment of the present invention, the wood is a leaf or stem of the wood.
본 발명의 바람직한 양태에 따르면, 상기 다래나무 에탄올 추출물에 대한 분획 용매로는 바람직하게는 앤-헥산(n-hexane), 클로로포름(Chloroform), 앤-부탄올(n-butanol) 및 에이퀴어스(Aqueous)를, 더욱 바람직하게는 앤-헥산(n-hexane), 앤-부탄올(n-butanol) 및 에이퀴어스(Aqueous)를, 가장 바람직하게는 에이퀴어스(Aqueous)를 사용할 수 있다.According to a preferred embodiment of the present invention, as the fraction solvent for the ethanol extract of the Methanol tree, preferably n-hexane, chloroform, n-butanol and Aqueous ), More preferably n-hexane, n-butanol and Aqueous, and most preferably Aqueous.
본 발명의 바람직한 양태에 따르면, 상기 분획물은 페놀계 화합물 또는 폴리 페놀계 화합물을 유효 성분으로 함유한다.According to a preferred embodiment of the present invention, the fraction contains a phenolic compound or a polyphenolic compound as an active ingredient.
본 발명의 바람직한 양태에 따르면, 상기 분획물은 플라보노이드계 화합물을 유효 성분으로 함유한다.According to a preferred embodiment of the present invention, the fraction contains a flavonoid compound as an active ingredient.
본 발명의 명세서에서 용어 ‘페놀(Phenol)’은 벤젠고리(C6H6)의 수소 중 1개가 하이드록시기(-OH)로 치환된 물질을 의미한다.In the present specification, the term “phenol” refers to a substance in which one of hydrogen in the benzene ring (C6H6) is substituted with a hydroxyl group (—OH).
본 발명의 명세서에서 용어 ‘폴리페놀(Polyphenol)’은 벤젠고리(C6H6)의 수소 중 2개 이상이 하이드록시기(-OH)로 치환된 물질을 의미한다.In the present specification, the term 'polyphenol' refers to a substance in which two or more hydrogens in the benzene ring (C6H6) are substituted with a hydroxyl group (—OH).
본 발명의 명세서에서 용어 ‘플라보노이드(Flavonoid)’는 식물에 널리 함유되어 있으며 2개의 페닐기가 피란 고리 혹은 그와 유사한 구조의 탄소원자 3개를 매개로 결합하고 있는 물질 군을 의미한다.In the present specification, the term 'flavonoid' refers to a group of substances widely contained in plants and in which two phenyl groups are bonded through three carbon atoms having a pyran ring or similar structure.
본 발명의 바람직한 양태에 따르면, 상기 분획물은 DPPH 라디칼 소거 능, ABTS 라디칼 소거 능 및 SOD 유사활성을 나타낸다.
According to a preferred embodiment of the present invention, the fraction exhibits DPPH radical scavenging ability, ABTS radical scavenging ability and SOD-like activity.
본 발명의 다른 양태에 따르면, 본 발명은 상술한 방법에 의해 수득한 상기 에탄올 추출물 또는 항산화 활성 분획 물을 유효 성분으로 함유하는 항산화용 약제학적 조성물, 항산화용 기능성 식품 조성물 및 항산화용 화장품 조성물을 제공한다.According to another aspect of the present invention, the present invention provides a pharmaceutical composition for antioxidant, functional food composition for antioxidant and cosmetic composition for antioxidant containing the ethanol extract or antioxidant active fraction obtained by the above method as an active ingredient. do.
본 발명의 바람직한 양태에 따르면, 상기 다래나무 에탄올 추출물 또는 항산화 활성 분획 물은 하기와 같은 특성을 가진다:According to a preferred embodiment of the present invention, the ethanol extract or the antioxidant active fraction of the cedar tree has the following characteristics:
(a) 총 페놀 함량(Total phenolic contents) : 탄닌산(Tannic acid) 40 내지 250±1 내지 5 mg/g;(a) Total phenolic contents: Tannic acid 40 to 250 ± 1 to 5 mg / g;
(b) 총 플라보노이드 함량(Total flavonid contents) : 퀘세틴(Quercetin) 10 내지 230±1 내지 4 mg/g;(b) Total flavonid contents: Quecetin 10-230 ± 1-4 mg / g;
(c) DPPH 라디칼 소거 능(DPPH radical scavenging activity) : 3 내지 60±0.1 내지 5%;(c) DPPH radical scavenging activity: 3 to 60 ± 0.1 to 5%;
(d) ABTS 라디칼 소거 능(ABTS radical scavenging activity) : 0.5 내지 100±0.1 내지 5%; 및(d) ABTS radical scavenging activity: 0.5-100 ± 0.1-5%; And
(e) SOD 유사활성(SOD-like activity) ; 2 내지 20±0.1 내지 12%.(e) SOD-like activity; 2-20 ± 0.1-12%.
본 발명에 있어서 상기 다래나무 에탄올 추출물 또는 항산화 활성 분획 물의 총 페놀 함량 및 총 플라보노이드 함량은 매우 중요하다. 왜냐하면, 페놀 계 화합물의 경우 생체의 노화 과정과 관련성이 있는 유리기 및 만성질환의 피해를 막아주는 산화 방지 역할을 하고 플라보노이드 계 화합물의 경우 항균, 항암, 항바이러스, 항 알레르기 및 항염증 활성을 나타내어 총 페놀 함량과 총 플라보노이드 함량이 높을수록 항산화 효능이 향상되기 때문이다.In the present invention, the total phenolic content and the total flavonoid content of the ethanol extract or antioxidant active fraction of Mt. Because phenolic compounds act as an antioxidant to prevent the damage of free radicals and chronic diseases related to the aging process of the body, and flavonoid compounds exhibit antimicrobial, anticancer, antiviral, antiallergic and anti-inflammatory activities. The higher the phenol content and the total flavonoid content, the better the antioxidant effect.
상기 다래나무 에탄올 추출물 또는 항산화 활성 분획 물의 수득 방법에 관해서는 상술한 내용과 중복되므로 본 명세서의 과도한 복잡성을 고려하여 생략하도록 한다.Regarding the method of obtaining the ethanol extract or the antioxidant active fraction of the cedar tree is duplicated as described above, so it will be omitted in consideration of the excessive complexity of the present specification.
본 발명의 바람직한 양태에 따르면, 본 발명의 조성물은 항산화 효과가 탁월한 화장료 조성물일 수 있다.According to a preferred embodiment of the present invention, the composition of the present invention may be a cosmetic composition excellent in antioxidant effect.
본 발명의 화장료 조성물은 상술한 다래 추출물 이외에 다른 성분들을 포함할 수 있다. 본 발명의 실시예에 따르면, 본 발명의 화장료 조성물은 글리세린, 부틸렌 글라이콜, 폴리옥시에칠렌 경화피마자유, 토코페릴 아세테이트, 시트릭산, 판테놀, 스쿠알란, 소듐 시트레이트 및 알란토인으로 포함된 군으로부터 선택되는 최소 하나의 보조성분을 추가적으로 포함하며, 보다 바람직하게는 글리세린, 폴리옥시에칠렌 경화피마자유, 토코페릴 아세테이트, 스쿠알란 및 소듐 시트레이트을 추가적으로 포함하고, 부틸렌 글라이콜, 시트릭산, 판테놀 및 알란토인으로 구성된 군으로부터 선택되는 최소 하나의 성분을 포함하며, 가장 바람직하게는 글리세린, 부틸렌 글라이콜, 폴리옥시에칠렌 경화피마자유, 토코페릴 아세테이트, 시트릭산, 판테놀, 스쿠알란, 소듐 시트레이트 및 알란토인 모두를 추가적으로 포함한다.The cosmetic composition of the present invention may include other components in addition to the above-mentioned Darae extract. According to an embodiment of the present invention, the cosmetic composition of the present invention is obtained from the group comprising glycerin, butylene glycol, polyoxyethylene hydrogenated castor oil, tocopheryl acetate, citric acid, panthenol, squalane, sodium citrate and allantoin Further comprising at least one auxiliary component selected from the group consisting of glycerin, polyoxyethylene hydrogenated castor oil, tocopheryl acetate, squalane and sodium citrate, and at least one auxiliary ingredient selected from the group consisting of butylene glycol, citric acid, panthenol and allantoin And most preferably at least one component selected from the group consisting of glycerin, butylene glycol, polyoxyethylene hydrogenated castor oil, tocopheryl acetate, citric acid, panthenol, squalane, sodium citrate and allantoin .
본 발명의 화장료 조성물은 기본적으로 피부에 도포되는 것이므로, 당업계의 화장료 조성물을 참조하여 제공될 수 있으며, 예를 들어, 용액, 현탁액, 유탁액, 페이스트, 겔, 크림, 로션, 파우더, 비누, 계면활성제-함유 클린싱, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션 및 스프레이 등으로 제형화될 수 있으나, 이에 한정되는 것은 아니다. 보다 상세하게는, 유연 화장수, 영양 화장수, 영양 크림, 마사지 크림, 에센스, 아이 크림, 클렌징 크림, 클렌징 포옴, 클렌징 워터, 팩, 스프레이 또는 파우더의 제형으로 제조될 수 있다.Since the cosmetic composition of the present invention is basically applied to the skin, it can be provided with reference to the cosmetic composition of the related art, for example, as a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, But are not limited to, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays. More specifically, it can be manufactured in the form of a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.
본 발명의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물성유, 식물성유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the formulation of the present invention is a paste, cream or gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .
본 발명의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄 또는 디메틸 에테르와 같은 추진체를 포함할 수 있다.In the case where the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. Especially, in the case of a spray, a mixture of chlorofluorohydrocarbons, propane / Propane or dimethyl ether.
본 발명의 제형이 용액 또는 유탁액인 경우에는 담체 성분으로서 용매, 용해화제 또는 유탁화제가 이용되고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌 글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르가 있다.When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.
본 발명의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올, 또는 프로필렌 글리콜과 같은 액상의 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가, 또는 트라칸트 등이 이용될 수 있다.When the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspension such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Crystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used.
본 발명의 제형이 계면-활성제 함유 클린징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르 설페이트, 설포숙신산 모노에스테르, 이세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사르코시네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산 디에탄올아미드, 식물성 유, 라놀린 유도체 또는 에톡실화 글리세롤 지방산 에스테르 등이 이용될 수 있다.When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, or ethoxylated glycerol fatty acid esters.
본 발명의 바람직한 양태에 따르면, 본 발명의 조성물은 항산화 효과가 탁월한 약제학적 조성물일 수 있다.According to a preferred embodiment of the present invention, the composition of the present invention may be a pharmaceutical composition having an excellent antioxidative effect.
본 발명의 조성물이 약제학적 조성물로 제조되는 경우, 본 발명의 약제학적 조성물은 약제학적으로 허용되는 담체를 포함한다. 본 발명의 약제학적 조성물에 포함되는 약제학적으로 허용되는 담체는 제제 시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. 본 발명의 약제학적 조성물은 상기 성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제등을 추가로 포함할 수 있다. 적합한 약제학적으로 허용되는 담체 및 제제는 Remington's Pharmaceutical Sciences (19th ed., 1995)에 상세히 기재되어 있다.When the composition of the present invention is manufactured from a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. The pharmaceutically acceptable carriers to be contained in the pharmaceutical composition of the present invention are those conventionally used in the formulation and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, But are not limited to, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrups, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. It is not. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).
본 발명의 약제학적 조성물은 쥐, 생쥐, 가축, 인간 등의 포유동물에 경구 또는 비경구 등의 다양한 경로로 투여할 수 있으며, 예컨대 경구, 직장 또는 정맥, 근육, 피하, 자궁내 경막 또는 뇌혈관내 주사에 의해 투여할 수 있다. 바람직하게는 비경구 투여 중 경피투여, 보다 바람직하게는 도포에 의한 국부 투여(topical application) 방식으로 적용된다.The pharmaceutical composition of the present invention can be administered to mammals such as rats, mice, livestock, and humans in various routes such as oral or parenteral routes such as oral, rectal or intravenous, muscular, subcutaneous, intra-uterine, Can be administered by injection. Preferably, it is applied by transdermal administration during parenteral administration, more preferably by topical application by application.
본 발명의 약제학적 조성물의 적합한 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하게 처방될 수 있다. 본 발명의 약제학적 조성물의 투여량은, 경구형 제형인 경우 성인 기준으로 0.1-100 ㎎/kg 의 양을 1일 1회 내지 수회 투여할 수 있으며, 외용제인 경우에는 성인 기준으로 1일당 1.0 내지 3.0 ml의 양으로 1일 1 내지 5회 도포하여 1개월 이상 계속 하는 것이 좋다. 다만, 상기 투여량은 본 발명의 범위를 한정하는 것은 아니다.The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate, . The dose of the pharmaceutical composition of the present invention can be administered in an amount of 0.1-100 mg / kg on an adult basis once or several times a day in the case of an oral formulation, and in the case of an external preparation, It is preferable to apply it once to 5 times a day in an amount of 3.0 ml and continue for 1 month or longer. However, the dosage is not intended to limit the scope of the present invention.
본 발명의 약제학적 조성물은 당해 발명이 속하는 기술분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있는 방법에 따라, 약제학적으로 허용되는 담체 및/또는 부형제를 이용하여 제제화함으로써 단위 용량 형태로 제조되거나 또는 다용량 용기 내에 내입시켜 제조될 수 있다. 이때 제형은 산제, 과립제, 정제, 캅셀제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 연고, 크림 등의 외용제, 좌제 및 멸균 주사용액 등을 비롯하여 약제학적 제제에 적합한 어떠한 형태로든 사용할 수 있으며, 분산제 또는 안정화제를 추가적으로 포함할 수 있다.The pharmaceutical compositions of the present invention may be prepared in unit dose form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporation into a multi-dose container. The formulations may be in any form suitable for pharmaceutical preparations including oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, external preparations such as ointments and creams, suppositories and sterile injectable solutions, , Dispersants, or stabilizers.
본 발명의 바람직한 양태에 따르면, 본 발명의 조성물은 항산화 효과가 탁월한 식품 조성물일 수 있다.According to a preferred embodiment of the present invention, the composition of the present invention may be a food composition having an excellent antioxidative effect.
본 발명의 조성물이 식품 조성물로 제조되는 경우, 유효성분뿐만 아니라, 식품 제조 시에 통상적으로 첨가되는 성분을 포함하며, 예를 들어, 단백질, 탄수화물, 지방, 영양소, 조미제 및 향미제를 포함한다. 상술한 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스, 올리고당 등; 및 폴리사카라이드, 예를 들어 덱스트린, 사이클로덱스트린 등과 같은 통상적인 당 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 향미제로서 천연 향미제 [타우마틴, 스테비아 추출물 (예를 들어 레바우디오시드 A, 글리시르히진 등]) 및 합성 향미제(사카린, 아스파르탐 등)를 사용할 수 있다.When the composition of the present invention is prepared with a food composition, it includes not only the active ingredient, but also ingredients normally added during the manufacture of the food, including, for example, proteins, carbohydrates, fats, nutrients, flavoring agents and flavoring agents . Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (saccharine, aspartame, etc.) can be used as flavorings.
예컨대, 본 발명의 식품 조성물이 드링크제로 제조되는 경우에는 본 발명의 유효성분 이외에 구연산, 액상과당, 설탕, 포도당, 초산, 사과산, 과즙, 두충 추출액, 대추 추출액, 감초 추출액 등을 추가로 포함시킬 수 있다.
For example, when the food composition of the present invention is prepared as a drink, it may further contain citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, mulberry extract, jujube extract, licorice extract, have.
한편, 본 발명의 구체적 실시예에서 본 발명 조성물의 유효성분에 대한 세포 생존율 분석 결과 본 발명 조성물의 유효성분은 천연물질로서 인체에 무해한 물질임이 밝혀졌다. 따라서, 본 발명의 유효성분은 독성 및 부작용이 거의 없으므로 장기간 사용 시에도 안심하고 사용할 수 있으며, 특히 상기한 바와 같은 화장료, 약제학적 및 식품 조성물에 안전하게 적용할 수 있다.On the other hand, cell specificity analysis of the active ingredient of the composition of the present invention in a specific embodiment of the present invention, it was found that the active ingredient of the composition of the present invention is a natural material harmless to the human body. Therefore, the active ingredient of the present invention has little toxicity and side effects, so that it can be used safely even in long-term use, and in particular, it can be safely applied to cosmetics, pharmaceutical and food compositions as described above.
본 발명의 특징 및 이점을 요약하면 다음과 같다: The features and advantages of the present invention are summarized as follows:
(a)본 발명은 다음 단계를 포함하는, 항산화 효능이 우수한 다래나무 추출물 및 분획 물 수득 방법을 제공한다: (ⅰ) 다래나무의 잎 또는 줄기의 분말에 70% 에탄올을 가하여 24시간동안 실온에서 진탕하여 2회 반복 추출한 후 여지로 여과한 뒤 농축기로 40℃에서 감압농축 및 동결건조 하여 70% 에탄올 추출물을 얻는 단계; (ⅱ) 상기 단계 (ⅰ)의 에탄올 추출물을 물에 용해시켜 분액깔때기에 넣은 뒤 분획 용매를 사용해 극성의 차이를 이용하여 분획 물을 얻는 단계를 포함시키는 다래나무 추출물 및 분획 물 수득법.(a) The present invention provides a method for obtaining antioxidant extract and fraction water having excellent antioxidant efficacy, comprising the following steps: (i) 70% ethanol is added to the powder of leaves or stems of a perilla tree at room temperature for 24 hours. Shaking and extracting twice, and then filtration with filtrate and concentrating under reduced pressure and freeze drying at 40 ° C. with a concentrator to obtain 70% ethanol extract; (Ii) the method of obtaining a mite tree extract and fraction water, comprising the step of dissolving the ethanol extract of step (iii) in water and placing it in a separatory funnel and using fractional solvent to obtain fraction water using a polarity difference.
(b) 본 발명의 다래나무 추출물 및 분획물은 항산화 관련 약제, 화장품 및 기능성 식품 조성물로 활용할 수 있어 발명의 활용 범위가 매우 넓다.(b) Since the extract and fractions of the present invention can be used as antioxidant-related drugs, cosmetics and functional food compositions, the scope of the invention is very wide.
(c) 본 발명의 다래나무는 생육 조건이 까다롭지 않고 한국의 풍토에 잘 서식하는 식물이기에 손쉽고 저렴하게 항산화 관련 제품 조성물의 원료를 대량 확보할 수 있다. 그리고 이는 추출물 및 분획 물을 활용한 제품의 가격을 낮추는 효과로 이어진다.(c) Since the present invention is not a demanding growth condition and a plant that lives well in the climate of Korea, it is possible to easily and inexpensively secure a large amount of raw material of antioxidant related product composition. This results in lowering the price of the product using the extract and the fraction.
(d) 본 발명의 다래나무의 추출물 및 분획 물 수득 공정이 까다롭지 않아 실 산업에의 적용이 매우 용이하며, 따라서 추출물 및 분획 물을 활용한 제품의 가격 역시 저렴하게 맞출 수 있다.(d) It is very easy to apply to the real industry because the extract and fraction water obtaining process of the mite tree of the present invention is not demanding, and thus the price of the product utilizing the extract and fraction water can also be adjusted inexpensively.
(e) 본 발명의 다래나무의 추출물 및 분획 물의 경우, 페놀 계 화합물 및 플라보노이드 계 화합물을 다량 함유하고 있어 항산화 효능이 뛰어나다. (e) The extracts and fractions of the stalks of the present invention contain a large amount of phenolic compounds and flavonoid compounds, which are excellent in antioxidant efficacy.
(f) 본 발명의 다래나무의 추출물 및 분획 물을 유효 성분으로 함유하는 항산화 관련 약제, 화장품 및 기능성 식품 조성물의 경우 인체에 어떠한 부작용도 일으키지 않는다.(f) Antioxidant-related agents, cosmetics and functional food compositions containing the extract and fraction of the present invention as an active ingredient do not cause any adverse effects on the human body.
도 1은 본 실험에서 다래나무의 잎 또는 줄기의 분말로부터 에탄올 추출물 및 항산화 활성 분획 물을 얻었던 과정을 모식 적으로 나타낸 것이다. 가장 상단의 Actinidia arguta Planchon는 다래나무의 잎 또는 줄기를 40 mesh로 마쇄한 분말을 의미한다. Vacuum evaporation은 감압 하에서 행하는 진공 증발을 의미한다. Freeze drying은 용액상태의 시료를 동결하여 그대로의 상태로 감압하에 방치함으로써 시료 중의 수분을 승화시켜 제거하는 동결건조를 의미한다. Etanol extract는 다래나무의 70% 에탄올 추출물을 의미한다. Etanol extract 하단의 과정은 다양한 종류의 용매에 의한 순차적 용매 분획 과정을 의미한다. Dissolve in water and extract with n-hexane은 상기 70% 에탄올 추출물을 물에 용해시켜 분액깔때기에 넣은 뒤 저 극성 용매인 n-hexane을 넣어 2 내지 4회 추출하여 n-hexane 분획 물을 얻는 것을 의미한다. 좌측의 n-hexane fraction이 n-hexane 분획 물을 의미한다. 우측의 Aqueous layer 및 그 하단의 과정들은 n-hexane 용매를 활용해서 n-hexane 분획 물을 얻고 난 이후의 수층을 다시 분액깔때기에 넣고 클로로포름(Chloroform)을 분획 용매로 하여 Chloroform 분획 물을 얻는 과정, 이후의 수층에 앤-부탄올(n-butanol) 분획 용매를 사용하여 n-butanol 분획 물을 얻는 과정 및 마지막에 남은 수층에서 Aqueous 분획 물을 얻는 과정을 의미한다.Figure 1 schematically shows the process of obtaining the ethanol extract and antioxidant active fractions from the powder of the leaves or stems of the wood in this experiment. Actinidia arguta Planchon at the top refers to a powder ground to 40 mesh of the leaves or stems of the stalk tree. Vacuum evaporation means vacuum evaporation performed under reduced pressure. Freeze drying refers to lyophilization, in which a sample in solution is frozen and left as it is under reduced pressure to sublimate and remove moisture in the sample. Etanol extract refers to 70% ethanol extract of the taraxacum. The process at the bottom of the ethanol extract refers to the sequential solvent fractionation process using various solvents. Dissolve in water and extract with n-hexane means that the 70% ethanol extract is dissolved in water and placed in a separatory funnel, followed by extraction 2 to 4 times with n-hexane, a low polar solvent, to obtain n-hexane fraction. . The n-hexane fraction on the left means the n-hexane fraction. The process of the Aqueous layer on the right side and the process below is to obtain the n-hexane fraction using n-hexane solvent, and then put the aqueous layer into the separatory funnel again to obtain the Chloroform fraction by using chloroform as the fraction solvent. It means the process of obtaining the n-butanol fraction in the subsequent aqueous layer using the n-butanol fraction solvent and the process of obtaining the Aqueous fraction in the last remaining aqueous layer.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명 하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로서, 본 발명의 요지 따라 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.
Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not to be construed as limiting the scope of the present invention. It will be self-evident.
실시예Example
실시예Example 1 : 추출물의 제조와 1: Preparation of extract 분획물에In the fraction 대한 분석 및 측정 For analysis and measurement
재료 및 방법Materials and methods
70% 에탄올 추출물 제조70% Ethanol Extract Preparation
40 mesh로 마쇄한 다래나무(Actinidia arguta Planchon) 분말 100 g에 10배량의 70% 에탄올을 가하여 24시간동안 실온에서 진탕하였다(Shaker SK-71 JEIO TECH, Korea). 2회 반복 추출한 후 여지(Advantec No. 6, Japan)로 여과한 뒤 농축기(EYELA N-3000, Japan)로 40℃에서 감압농축 및 동결건조 하여 70% 에탄올 추출물을 얻었다.
Wood crushed with 40 mesh ( Actinidia) arguta Planchon ) was added to 10 g of 70% ethanol and shaken at room temperature for 24 hours (Shaker SK-71 JEIO TECH, Korea). After repeated extraction twice, the resultant was filtered through a filter paper (Advantec No. 6, Japan) and concentrated under reduced pressure and freeze-dried at 40 ° C. with a concentrator (EYELA N-3000, Japan) to obtain 70% ethanol extract.
70% 에탄올 추출물을 이용한 분획제조Fraction Production with 70% Ethanol Extract
순차적 용매 분획은 상기 70% 에탄올 추출물 10 g을 물 500 mL에 용해시켜 분액깔때기에 넣은 뒤 저 극성 용매인 n-hexane 500 mL을 넣어 3회 추출하여 n-hexane 분획 물을 얻었다. 그 수층을 다시 분액깔때기에 넣고 위와 같은 방법으로 클로로포름(Chloroform), 앤-부탄올(n-butanol) 및 에이퀴어스(Aqueous)의 순서로 극성의 차이를 이용하여 분획 물을 얻었다(도 1). 분리된 각각의 용매 분획 물은 감압 농축하여 용매를 제거 한 후 -70℃에서 동결 건조하여 각 분획의 수율을 구하고, 일정량을 취하여 항산화 활성 분석용 시료로 사용 하였다.
The sequential solvent fraction was dissolved 10 g of the 70% ethanol extract in 500 mL of water and placed in a separatory funnel, and then extracted three times by adding 500 mL of n-hexane, a low polar solvent, to obtain n-hexane fraction. The aqueous layer was placed in a separatory funnel again to obtain fractions using the difference in polarity in the order of chloroform, n-butanol and Aqueous in the same manner as above (FIG. 1). The separated solvent fractions were concentrated under reduced pressure to remove the solvent, and then freeze-dried at -70 ° C to obtain the yield of each fraction, and a certain amount was taken as an antioxidant activity assay sample.
총 폴리페놀 함량 분석Total Polyphenol Content Analysis
1 mg/mL의 농도로 용해한 각 용매별 분획 물 10 ㎕에 폴린-시오칼토(Folin-Ciocalteau)시약 10 ㎕ 및 2% 탄산나트륨(Na2CO3) 200 ㎕를 차례로 가한 다음 실온에서 30분간 정치한 후 750 nm에서 흡광도를 측정하였다. 총 폴리페놀 함량은 탄닌산(Tannic acid)을 표준물질로 정하여 시료와 동일한 방법으로 분석하여 작성한 검량 선으로부터 계산하였다.
10 μl of Folin-Ciocalteau reagent and 200 μl of 2% sodium carbonate (Na2CO3) were added sequentially to 10 μl of each solvent dissolved at a concentration of 1 mg / mL, and then allowed to stand at room temperature for 30 minutes, followed by 750 nm. Absorbance was measured at. The total polyphenol content was calculated from the calibration curve prepared by analyzing tannic acid as a standard and analyzing it in the same manner as the sample.
총 플라보노이드 함량Total flavonoid content
총 플라보노이드 함량은 용매별 분획 물 20 ㎕를 아질산나트륨 용액(Sodium nitrite solution) 75 ㎕와 혼합하여 5분간 반응시키고 염화알루미늄 용액(Aluminum chloride solution) 150 ㎕를 첨가하여 다시 5분간 반응시킨 후 1 M 수산화나트륨(NaOH) 20 ㎕와 섞어 510 nm에서 흡광도를 측정하였다. 총 플라보노이드 함량은 쿼세틴(Quercetin)을 이용하여 작성한 표준곡선으로부터 함량을 구하였다.
The total flavonoid content was mixed with 75 μl of sodium nitrite solution for 5 minutes, and then reacted for 5 minutes by adding 150 μl of aluminum chloride solution. The absorbance was measured at 510 nm with 20 μl of sodium (NaOH). The total flavonoid content was determined from the standard curve prepared using quercetin.
DPPHDPPH 라디칼Radical 소거 능 측정 Emissivity measurement
DPPH 라디칼에 대한 소거 능은 각 용매별 분획 물 40 ㎕와 1,1-diphenyl-2-picryhydrazyl (DPPH)용액 160 ㎕를 가하여 잘 혼합한 후 암소에서 30분간 반응시킨 후 518 nm에서 흡광도를 측정하여 농도에 따른 DPPH 라디칼 소거 능을 확인하였다. 시료 무 첨가구와 비교하여 자유라디칼 소거활성을 백분율로 나타냈다.
For the scavenging ability of DPPH radicals, 40 μl of each fraction and 160 μl of 1,1-diphenyl-2-picryhydrazyl (DPPH) solution were added, mixed well, and reacted in the dark for 30 minutes, and then absorbance was measured at 518 nm. DPPH radical scavenging ability according to the concentration was confirmed. The free radical scavenging activity was expressed as a percentage compared to the sample free addition.
ABTSABTS 라디칼Radical 소거능Scatters 측정 Measure
ABTS［2,2'-azin0-bis(3-ethylbenzthiazoline-6-sulfonate)］라디칼 소거 능은 과 황산칼륨(Potassium persulfate)과의 반응에 의해 생성된 ABTS 유리 라디칼이 시료의 항산화 물질에 의해 제거되어 라디칼 특유의 색인 청록색이 탈색되는 것을 이용한 방법으로 측정하였다. 7.2 mM ABTS용액에 2.6 mM 과 황산칼륨을 혼합시킨 다음 암실에서 약 15시간 반응시켰다. 이를 734 nm에서 흡광도가 1.5가 되도록 희석한 후 300 ㎕를 취하여 용매별 분획 물 20 ㎕를 혼합하여 암소에서 30분간 반응시켜 734 nm에서 흡광도를 측정하여 시료첨가구와 무 첨가구의 흡광도 차이를 백분율로 나타내었다.
ABTS [2,2'-azin0-bis (3-ethylbenzthiazoline-6-sulfonate)] radical scavenging ability is due to the removal of ABTS free radicals produced by the reaction with potassium persulfate. It measured by the method using the thing of the radical blue-green color discoloring. 2.6 mM ABTS solution was mixed with 2.6 mM potassium sulfate and reacted in the dark for about 15 hours. After diluting the absorbance to 1.5 at 734 nm, take 300 µl, mix 20 µl of the fractions for each solvent, react for 30 minutes in the dark, and measure the absorbance at 734 nm. It was.
SODSOD 유사활성Similar activity 측정 Measure
용매별 분획물의 SOD유사활성은 과산화수소(H2O2)로 전환시키는 반응을 촉매하는 피로갈롤(Pyrogallol)의 생성량을 측정하여 나타내었다. 용매별 추출물 20 ㎕에 트리스-염화수소 완충액(Tris-Hcl buffer, 50 mM tris (hydroxymethyl) amino-methane + 10 mM EDTA, pH 8.5)을 이용하여 pH 8.5로 조절된 시료 액을 만들어 시료 액 260 ㎕와 7.2 mM 피로갈롤 20 ㎕를 가하고 상온에서 10분간 방치 한 후 1N 염화수소 10 ㎕로 반응을 정지시킨 후 산화된 피로갈롤의 흡광도를 420 nm에서 측정하였다. SOD 유사활성은 시료 첨가구 및 무 첨가구의 흡광도 차이를 백분율로 나타내었다.
The SOD-like activity of the fractions by solvent was measured by measuring the amount of pyrogallol that catalyzes the conversion of hydrogen peroxide (H 2 O 2). 20 μl of the extract for each solvent was used to make a sample solution adjusted to pH 8.5 using Tris-Hcl buffer, 50 mM tris (hydroxymethyl) amino-methane + 10 mM EDTA, pH 8.5. 20 μl of 7.2 mM pyrogallol was added thereto, and the mixture was left at room temperature for 10 minutes and the reaction was stopped with 10 μl of 1N hydrogen chloride. Then, the absorbance of the oxidized pyrogallol was measured at 420 nm. SOD-like activity was expressed as a percentage difference in absorbance between the sample addition group and no addition group.
결과 및 고찰Results and Discussion
다래의 용매분획별 추출수율은 표 1에서와 같이 극성이 높은 수용액 분획 물에서 가장 높은 수율을 보였고, 극성이 낮은 클로로포름 분획 물에서 가장 낮았다. The extraction yield of each solvent fraction showed the highest yield in the aqueous solution fraction of high polarity as shown in Table 1, and the lowest in the chloroform fraction of low polarity.
총 페놀과 총 플라보노이드(Flavonoid) 함량결과는 표 2에서 보이는 바와 같이 플라보노이드 배당체의 대다수가 부탄올 분획 물 층으로 이동하였기에 총 페놀 및 총 플라보노이드 함량이 다른 분획층보다 부탄올 분획 층에서 높게 나타났다.The total phenol and flavonoid content results showed that the majority of flavonoid glycosides were transferred to the butanol fraction layer as shown in Table 2, so the total phenol and total flavonoid contents were higher in the butanol fraction layer than in the other fraction layers.
다래 분획물의 DPPH 라디칼 소거 능은 표 3에서 보이는 바와 같이 천연항산화제보다 높게 나타나지 않았지만 농도 의존적인 활성을 보였다. 특히 100 ㎍/mL 농도에서의 부탄올 분획 물 층은 합성 항산화제인 BHT와 천연항산화제인 알파-토코페롤(α-tocopherol) 보다는 높은 DPPH 라디칼 소거 능을 보였다. DPPH radical scavenging ability of the stalk fraction was not higher than that of natural antioxidants as shown in Table 3, but showed concentration-dependent activity. In particular, the butanol fraction layer at the concentration of 100 ㎍ / mL showed higher DPPH radical scavenging ability than the synthetic antioxidant BHT and the natural antioxidant alpha-tocopherol.
다래 분획물의 ABTS 라디칼 소거 능은 표 4에서 보이는 바와 같이 천연항산화제보다 높게 나타나지 않았지만 농도 의존적인 활성을 보였다. 특히 500 ㎍/mL 농도에서의 부탄롱 분획 물 층은 합성 항산화제인 BHT와 천연항산화제인 알파-토코페롤 및 엘-아스코르빈산(L-ascorbic acid)보다는 높은 ABTS 라디칼 소거 능을 보였다.The ABTS radical scavenging ability of the stalk fraction was not higher than that of the natural antioxidants as shown in Table 4, but showed concentration-dependent activity. In particular, the butanelong fractional layer at 500 ㎍ / mL showed higher ABTS radical scavenging ability than synthetic antioxidant BHT and natural antioxidants alpha-tocopherol and L-ascorbic acid.
다래 분획물의 SOD 유사활성은 다른 항산화활성지표인 DPPH, ABTS 라디칼 소거 능보다는 높은 활성을 보이지 않았으나 농도 의존적으로 활성을 보였다(표 5). SOD-like activity of the Darae fraction showed no higher activity than DPPH and ABTS radical scavenging ability, which were other antioxidant activities, but showed concentration-dependent activity (Table 5).
Claims (8)
Ethanol extract of Actinidia arguta Planchon with antioxidant activity.
The extract of claim 1, wherein the wood is a leaf or stem of a wood.
The method of claim 1, wherein the ethanol extract of the Methanol tree is selected from the group consisting of n-hexane, chloroform, n-butanol and Aqueous Extract, characterized in that the antioxidant active fraction (Fraction) of the selected solvent.
[화학식 1]
The extract as claimed in claim 3, wherein the fraction contains a flavonoid compound having a structure of Formula 1 as an active ingredient.
[Formula 1]
[화학식 2] [화학식 3]
4. The extract of claim 3, wherein the fraction contains a phenol-based compound having a structure of Formula 2 or Formula 3 as an active ingredient.
[Chemical Formula 2] < EMI ID =
The extract according to claim 3, wherein the fraction exhibits DPPH radical scavenging ability, ABTS radical scavenging ability and SOD-like activity.
The pharmaceutical composition for antioxidant, the functional food composition for antioxidant, and the cosmetic composition for antioxidant containing the said extract as described in any one of Claims 1-6 as an active ingredient.
(a) 상기 추출물의 총 페놀 함량(Total phenolic contents) : 탄닌산(Tannic acid) 40 내지 250±1 내지 5 mg/g;
(b) 상기 추출물의 총 플라보노이드 함량(Total flavonid contents) : 퀘세틴(Quercetin) 10 내지 230±1 내지 4 mg/g;
(c) 상기 추출물의 DPPH 라디칼 소거 능(DPPH radical scavenging activity) : 3 내지 60±0.1 내지 5%;
(d) 상기 추출물의 ABTS 라디칼 소거 능(ABTS radical scavenging activity) : 0.5 내지 100±0.1 내지 5%; 및
(e) 상기 추출물의 SOD 유사활성(SOD-like activity) ; 2 내지 20±0.1 내지 12%.Ethanol extract of Actinidia arguta Planchon with antioxidant activity, comprising the following properties:
(a) Total phenolic contents of the extract: Tannic acid 40 to 250 ± 1 to 5 mg / g;
(b) Total flavonid contents of the extract (Total flavonid contents): Quecetin 10 to 230 ± 1 to 4 mg / g;
(c) DPPH radical scavenging activity of the extract: 3 to 60 ± 0.1 to 5%;
(d) ABTS radical scavenging activity of the extract: 0.5-100 ± 0.1-5%; And
(e) SOD-like activity of the extract; 2-20 ± 0.1-12%.
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