KR101848252B1 - A composition comprising the Leukodin isolated from the extract of Artemisia Capillaris for skin whitening - Google Patents
A composition comprising the Leukodin isolated from the extract of Artemisia Capillaris for skin whitening Download PDFInfo
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- KR101848252B1 KR101848252B1 KR1020160129178A KR20160129178A KR101848252B1 KR 101848252 B1 KR101848252 B1 KR 101848252B1 KR 1020160129178 A KR1020160129178 A KR 1020160129178A KR 20160129178 A KR20160129178 A KR 20160129178A KR 101848252 B1 KR101848252 B1 KR 101848252B1
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- South Korea
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- composition
- extract
- artemisia
- leukodin
- skin
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Abstract
Description
본 발명은 인진쑥(Artemisia Capillaris) 추출물로부터 분리된 류코딘(Leukodin)을 유효성분으로 함유하는 피부 미백용 조성물에 관한 것으로서, 더욱 상세하게는 미백 활성이 증진된 인진쑥 추출물 또는 이의 분획물의 제조방법에 관한 것이다. The present invention relates to the use of Artemisia The present invention relates to a composition for skin whitening comprising Leukodin (Leukodin) isolated from a Capillaris extract as an active ingredient. More particularly, the present invention relates to a composition for enhancing whitening activity or a method for producing a fraction thereof.
사람의 피부는 노화 과정에서 다양한 물리, 화학적 변화가 일어나는데, 다양한 광화학반응에 의한 피부장애를 최소한으로 방지하기 위한 작용을 갖고 있다. 멜라닌 색소, SOD(superoxide dismutase) 및 그 외의 항산화 성분 등은 피부 내부에 침투한 자외선을 흡수하고, 그 에너지를 감쇄시키거나 자외선이 이차적으로 발생시키는 활성산소를 소거함으로써 피부에 대한 장를 방어하는데, 그 중 멜라닌은 자외선을 차단하여 진피 이하의 피부 기관을 보호해 주는 동시에 피부의 생체 내 생겨난 자유라디칼 등에 의한 피부 내 단백질과 유전자들의 손상을 보호해 주는 유용한 역할을 한다. 그러나 멜라닌이 필요 이상으로 많이 생성되면 피부 노화를 가져오며 기미나 주근깨 등과 같은 과색소 침착증을 유발한다. 멜라닌은 활성 산소종을 제거하는 기능이 탁월하나, 때로는 멜라닌 자체가 활성산소를 발생시키기도 하며 멜라닌 구조 내 카테콜이나 퀴논에 의하여 다른 물질을 환원시키거나 산화시키고 멜라닌 자체가 자유라디칼의 성질을 나타내기도 한다. 따라서 멜라닌 생성을 저해시키면 피부 미백 효과를 가져 올 수 있다. 멜라닌 생성을 저해시킴으로써 피부 미백 효과를 갖는 것으로, 파라-메톡시페놀(p-metoxyphenol), 하이드로퀴논(hydroquinone), 코지산(kojic acid), 알부틴(albutin), 비타민 C(ascorbic acid), 글루타티온(glutathione), 시스테인(cystein) 등의 화합물과, 상백피 추출물, 감초 추출물, 작약 추출물, 계피 추출물, 고삼 추출물, 갈근 추출물, 당귀 추출물, 목단피 추출물, 반하 추출물, 알로에 추출물 등의 식물 추출물 등이 알려져 있으나, 불충분한 미백 효과, 피부에 대한 안전성 문제, 화장료에 배합시 제형 내에서의 안정성 문제 등 여전히 해결해야 하는 문제점을 안고 있다.Human skin has various physical and chemical changes in the aging process, and it has an action to minimize the skin disorder caused by various photochemical reactions. Melanin pigment, superoxide dismutase (SOD), and other antioxidant ingredients absorb the ultraviolet rays penetrating the inside of the skin and defend the skin against the skin by attenuating the energy or erasing the secondary oxygen generated by the ultraviolet rays. Heavy melanin protects the subcutaneous tissue of the skin by blocking ultraviolet light, while protecting skin from damaging free radicals and other proteins and genes in the skin. However, when melanin is produced more than necessary, it causes skin aging and hypercholesterolemia such as spots and freckles. Melanin has the ability to remove reactive oxygen species, but sometimes melanin itself generates free radicals. It also reduces or oxidizes other substances by catechol or quinone in the melanin structure, and melanin itself has the property of free radicals do. Therefore, inhibition of melanin production may result in a skin whitening effect. (P-methoxyphenol), hydroquinone, kojic acid, albutin, vitamin C (ascorbic acid), glutathione (glutathione) and the like, which have a skin whitening effect by inhibiting melanin production. glutathione and cystein and plant extracts such as extracts of cucumber herb extract, licorice extract, peony extract, cinnamon extract, gossam extract, Puerariae Radix extract, Angelica gigantosa extract, Problems such as insufficient whitening effect, safety to skin, stability in formulations when added to cosmetics, and the like are still problems to be solved.
쑥(Artemisia)은 한국을 비롯한 아시아와 유럽 등에 분포되어 자생하는 국화과(compositae)의 다년생 초본 식물로서 약 400여종이 존재하는 것으로 알려져 있고 약 300여종이 우리나라에 자생하고 있으며, 그 중에서 한국 등록특허 제10-0401472호(인진쑥 추출물 함유 기능성 음료 및 그 제조방법)에서는 혈관이완용, 뇌졸중예방용 또는 항암성 음료 용도의 인진쑥 추출물을 기재하고 있다. 쑥과 같은 천연물을 대상으로 용매 극성을 이용하여 분획물을 얻는 방법은 통상적으로 헥산, 클로로포름, 디클로로메탄, 에틸아세테이트, 부탄올을 순차적으로 처리하여 극성에 따라 분획물을 세분화하는 것이다. 하지만, 이러한 방법은 극성에 따라 물질 분리를 세분화할 수 있는 장점이 있지만, 활성성분을 분리하는 실험에 있어서는 활성성분이 분산되는 현상이 일어난다.Artemisia is a perennial herbaceous plant of Compositae which is distributed in Korea and Asia and Europe. It is known that there are about 400 species. About 300 species are native to Korea, 10-0401472 (Functional beverage containing Artemisia sp. Extract and method for producing the same) describes Artemisia sp. Extract for use in blood vessel relaxation, stroke prevention, or anticancer beverage. A method for obtaining fractions using solvent polarity for natural products such as mugwort is to sequentially treat hexane, chloroform, dichloromethane, ethyl acetate, and butanol to fractionate the fractions according to the polarity. However, this method has an advantage of being capable of subdividing the substance according to the polarity, but in the experiment for separating the active ingredient, the active ingredient is dispersed.
이에, 본 발명자들은 기존 알려진 용매 분획 방법에 비해, 활성성분의 분산을 방지하고 더 많은 양의 활성 분획물을 얻을 수 있는 방법을 고안하고 이를 인진쑥 추출물에 적용함으로써, 적은 양으로도 피부 미백 활성을 가장 잘 나타낼 수 있는 인진쑥 추출물 또는 인진쑥 추출물로부터 분리된 류코딘(Leukodin)을 유효성분으로 함유하는 피부 미백용 조성물을 제조하는 방법을 개발하고자 하였다. Accordingly, the inventors of the present invention have devised a method of preventing the dispersion of the active ingredient and obtaining a larger amount of the active fraction as compared with the known solvent fractionation method, (Leukodin) as an active ingredient, which is isolated from Artemisia sp. Extract or Artemisia sp.
본 발명의 목적은, 유기용매를 이용하여 류코딘(Leukodin)을 유효성분으로 함유하는 인진쑥 분획물 및 이의 제조 방법을 제공함으로서, 인체에 무해하면서 피부 미백에 효과가 있는 화장료 조성물, 비누 조성물, 약제학적 조성물 및 식품 조성물 등을 제공함에 있다.It is an object of the present invention to provide an artificial broad bean fraction containing Leukodin as an active ingredient using an organic solvent and a method for producing the same, thereby providing a cosmetic composition, a soap composition, a pharmaceutical composition Compositions, food compositions and the like.
상기 목적을 달성하기 위하여, 본 발명은 인진쑥 추출물로부터 분리된 류코딘(Leukodin)을 유효성분으로 함유하는 피부 미백용 조성물을 제공한다.In order to achieve the above object, the present invention provides a skin whitening composition comprising Leukodin isolated from Artemisia sp. Extract as an active ingredient.
상기 조성물은 멜라닌 생성을 억제하는 것을 특징으로 하며, 화장료 조성물, 비누 조성물, 약학적 조성물, 식품 조성물인 것을 특징으로 한다.The composition is characterized by inhibiting melanin production, and is characterized by being a cosmetic composition, a soap composition, a pharmaceutical composition, and a food composition.
상기 본 발명은 인진쑥 추출물로부터 분리된 류코딘(Leukodin)을 유효성분으로 함유하는 피부 미백용 조성물의 제조 방법으로서, (1) 인진쑥(Artemisia Capillaris)에 물, 에탄올 또는 이들의 혼합용매로 인진쑥 추출물을 제조하는 단계; (2) 상기 인진쑥 추출물을 유기용매로 분획하여 인진쑥 분획물을 제조하는 단계; 를 단계를 포함하는 것을 다른 특징으로 한다.The present invention relates to a method for producing a skin whitening composition comprising Leukodin isolated from an extract of Artemisia japonica as an active ingredient, which comprises (1) Artemisia Capillaris ) with water, ethanol or a mixed solvent thereof; (2) fractionating the mugwort water extract with an organic solvent to prepare an mugwort fraction; The method comprising the steps of:
상기 (2)단계의 유기용매는 에틸아세테이트, 메탄올, 헥산 또는 이들의 혼합 용액인 것을 특징으로 한다. The organic solvent in step (2) is ethyl acetate, methanol, hexane or a mixture thereof.
상기 제조 방법에 있어서, 제조된 인진쑥 분획물을 분리 정제하는 단계를 더 포함할 수 있다.In the above production method, it may further comprise the step of separating and purifying the produced Artemisia spp. Fraction.
상기와 같은 본 발명에 따르면, 피부 미백 활성을 갖는 인진쑥 추출물 또는 피부 미백 활성이 증진된 인진쑥 추출물의 제조방법을 제공함으로써, 피부에 안전하면서도 피부 과색소 침착 질환을 예방 또는 개선하는 효과가 있다.According to the present invention, there is provided a method for manufacturing an extract of Artemisia syrup extract having skin whitening activity or skin whitening activity with skin whitening activity, thereby preventing or ameliorating skin hyperpigmentation diseases while being safe for skin.
도 1은 본 발명의 일 실시예에 따른 인진쑥 추출물 또는 인진쑥 분획물의 제조 공정을 나타내는 모식도이다.
도 2는 본 발명의 일 실시예에 따른 MPLC (medium pressure liquid chromatography)분획 결과이다.
도 3은 본 발명의 일 실시예에 따른 인진쑥 분획물(MPLC fraction 3)로부터 prep-HPLC로 분리 정제한 결과이다.
도 4는 본 발명의 일 실시예에 따라 인진쑥 분획물에서 분리 정제된 류코딘(Leukodin)을 HPCL(High Performance liquid chromatography)로 분석한 결과이다.
도 5는 본 발명의 일 실시예에 따른 류코딘(Leukodin)의 수소핵자기공명(1H NMR, 1H Nuclear Magnetic Resonance)의 분석 결과이다.
도 6은 본 발명의 일 실시예에 따른 류코딘(Leukodin)의 탄소핵자기공명(13C NMR, 13C Nuclear Magnetic Resonance)의 분석 결과이다.
도 7은 본 발명의 일 실시예에 따른 류코딘(Leukodin)의 세포 내 멜라닌 생성 억제 활성을 측정한 사진이다.
도 8은 본 발명의 일 실시예에 따른 류코딘(Leukodin)의 세포 내 멜라닌 생성 억제 활성을 측정한 결과 그래프이다.
도 9는 본 발명의 일 실시예에 따른 류코딘(Leukodin)의 세포 독성을 측정한 결과 그래프이다.FIG. 1 is a schematic view showing a process for producing an extract of Artemisia princeps or Artemisia sp., According to an embodiment of the present invention.
FIG. 2 is a result of a medium pressure liquid chromatography (MPLC) fraction according to an embodiment of the present invention.
FIG. 3 shows the result of purification by prep-HPLC from an artificial feces fraction (MPLC fraction 3) according to an embodiment of the present invention.
FIG. 4 shows the results of HPLC analysis of leukodin (Leukodin) isolated from Artemisia sp. Fractions according to an embodiment of the present invention.
Figure 5 shows the results of analysis of hydrogen nuclear magnetic resonance ( 1 H NMR, 1 H Nuclear Magnetic Resonance) of Leukodin according to an embodiment of the present invention.
FIG. 6 shows the results of analysis of carbon nuclear magnetic resonance ( 13 C NMR, 13 C nuclear magnetic resonance) of Leukodin according to an embodiment of the present invention.
FIG. 7 is a photograph showing the activity of inhibiting intracellular melanin formation of leucodin according to an embodiment of the present invention. FIG.
FIG. 8 is a graph showing the results of measuring the inhibitory activity of leucodin in cells for producing melanin according to an embodiment of the present invention.
FIG. 9 is a graph showing cytotoxicity of leucodin according to an embodiment of the present invention. FIG.
이하, 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명의 일 실시예에 의한 인진쑥 추출물로부터 분리된 류코딘(Leukodin)을 유효성분으로 함유하는 피부 미백용 조성물을 제공한다. There is provided a skin whitening composition comprising Leukodin isolated from Artemisia sp. Extract as an active ingredient according to an embodiment of the present invention.
국화과 쑥속에 속하는 인진쑥은 국내 각지에서 자생하는 사철쑥(Artemisia capillaris; AC)으로서 그 어린 줄기와 잎을 인진호(茵陳蒿)라 하고, 이담작용, 천식억제, 구충작용, 이뇨작용 등이 알려져 있으며 그 성분으로는 스코파론(Scoparone), 클로로겐산(Chlorogenic acid), 카필린(Capillin) 등이 알려져 있다(정보섭 및 신민교, 향약대사전, pp 1016-1017, 영림사, 1998). 국화과 쑥속에 속하는 다년생초는 아래와 같이 여러 종이 있으며, 같은 속에 속한다 하더라도 그 특징과 함유된 성분, 활성물질의 차이로 인해 각기 다른 효능을 나타낸다. Artemisia spp., Which is a member of the mugwort, capillaris ; AC), and its young stem and leaves are known as in Jin Chen (阴 Chen 蒿), and it is known that dandruff, asthma suppression, anthelmintic effect, diuretic effect and the like include scoparone, chlorogenic acid, Capillin, etc. are known (Information Supervision and Shin Min-gyo, Encyclopedia of Enlightenment, pp 1016-1017, Young Lim, 1998). A perennial herb belonging to Asteraceae Mugwort is several species as follows. Even if they belong to the same genus, they exhibit different effects due to differences in their characteristics, contained components and active substances.
본 발명의 일 실시예에 따른 조성물은 인진쑥 추출물 또는 이의 분획물이 인체에 독성을 미치지 않는 범위 내에서 임의의 양으로 포함될 수 있고 바람직하게는 조성물 전체 중량에 대하여 0.001 내지 99.99 중량%로 포함될 수 있다. The composition according to an embodiment of the present invention may be contained in any amount within a range that the extract of Artemisia princeps or its fraction is not toxic to the human body, and may be contained in an amount of 0.001 to 99.99% by weight based on the total weight of the composition.
본 실시예에 따른 인진쑥 추출물로부터 분리된 류코딘(Leukodin)을 유효성분으로 함유하는 피부 미백용 조성물에 있어서, 류코딘(Leukodin)은 하기 [화학식 1]로 표시될 수 있으며, 멜라닌의 생성을 억제할 수 있고, 피부 잡티를 제거하는 미백효과가 있어 맑고 건강한 피부를 유지시켜 줄 수 있다.Leukodin can be represented by the following formula (1), and inhibits the production of melanin in skin whitening compositions containing leucodin (Leukodin) isolated from Artemisia sp. Extract according to this embodiment as an active ingredient And it has a whitening effect that removes skin roughness, and can maintain a clear healthy skin.
[화학식 1][Chemical Formula 1]
본 실시예에 따른 상기 조성물은 화장료 조성물로 이용될 있다. 에멀젼, 로션, 크림(수중유적형, 유중수적형, 다중상), 용액, 현탁액(무수 및 수계), 무수 합성물(오일 및 글리콜계), 젤, 마스크, 팩 또는 분말 등의 형태로 제조될 수 있고, 화장품 제제에 있어서 당해 기술분야에서 통상의 기술자가 적절하게 선택하여 사용 가능한 알코올, 오일, 계면 활성제, 지방산, 실리콘 오일, 습윤제, 보습제, 점성 변형제, 유제, 안정제, 자외선 차단제, 발색제, 향료 등의 담체를 포함할 수 있다.The composition according to this embodiment may be used as a cosmetic composition. Can be prepared in the form of emulsions, lotions, creams (oil-in-water type, multimix), solutions, suspensions (anhydrous and water), anhydrous compounds (oil and glycol) In the cosmetic preparations, there can be used alcohol, oil, surfactant, fatty acid, silicone oil, wetting agent, humectant, viscous modifier, emulsifier, stabilizer, sunscreen agent, coloring agent, perfume And the like.
본 실시예에 따른 상기 조성물은 비누 조성물로 이용될 수 있다. 야자유, 팜유, 대두유, 파마자유, 올리브유, 팜핵류 등의 식물유지 또는 우지, 돈지, 양지, 어유 등의 동물유지 등의 비누 베이스에 첨가제로서 피부 보습제, 유화제, 경수연화제 등을 포함하여 제조될 수 있다. 상기 피부 보습제로서는 글리세린, 에리트리톨, 폴리에틸렌글리콜, 프로필렌글리콜, 부틸렌글리콜, 펜틸렌글리콜, 헥실글리콜, 이소프로필미리스테이트, 실리콘 유도체, 알로에베라, 솔비톨 등이 사용될 수 있으며, 상기 유화제로서는 천연오일, 왁스 지방알콜, 탄화수소류, 천연식물 추출물 등이 사용될 수 있고, 상기 경수연화제로서는 테트라소듐 이디티에이 등이 사용될 수 있다. 또한, 첨가제로서 항균제, 분산제, 거품억제제, 용매, 물때 방지제, 부식 방지제, 향료, 색소, 금속 이온봉쇄제, 산화방지제, 방부제 등을 추가적으로 포함할 수 있다.The composition according to this embodiment can be used as a soap composition. Emulsifying agent, water softening agent, etc. as an additive to a soap base such as palm oil, palm oil, soybean oil, pharmacopoeial, olive oil and palm kernel oil, or animal fats such as tallow, lard, have. As the above-mentioned skin moisturizing agent, glycerin, erythritol, polyethylene glycol, propylene glycol, butylene glycol, pentylene glycol, hexyl glycol, isopropyl myristate, silicone derivatives, aloe vera, sorbitol, Wax fatty alcohols, hydrocarbons, natural plant extracts and the like can be used. As the water softening agent, tetrasodium edetate and the like can be used. The additive may further include an antimicrobial agent, a dispersant, a foam inhibitor, a solvent, a scouring inhibitor, a corrosion inhibitor, a perfume, a dye, a sequestering agent, an antioxidant, an antiseptic and the like.
본 실시예에 따른 상기 조성물은 약제학적 조성물로 이용될 수 있다. 구체적으로 염증 반응에서 비롯되는 여드름, 아토피, 건선, 피부염 등과, 활성산소 증가에서 비롯되는 피부 노화, 주름 등과, 멜라닌 색소의 합성 증가로 피부에 국소적으로 발생하는 기미, 주근깨, 검버섯, 모반, 흑색종, 약물에 의한 색소 침착, 염증에 의한 색소 침착 등과 같은 피부 색소 침착 질환에 대해 예방 또는 치료 효과가 있는 약제학적 조성물로 이용이 가능하다. 약제학적 조성물은 임상 투여 시에 경구, 경피, 피하, 정맥 또는 근육을 포함한 여러 경로를 통해 투여될 수 있고 피부 외용제로 도포될 수 있다. 또한, 여러 가지 제형으로 제제화할 수 있는데, 제제화할 경우에는 통상적으로 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제 및 계면활성제 등의 희석제 또는 부형제를 사용하여 제제화할 수 있다. 경구 투여를 위한 고형 제제에는 정제, 환제, 산제, 과립제 및 캡슐제 등이 포함되며, 이러한 고형 제제는 콩 추출물 등에 적어도 하나 이상의 부형제, 예를 들면 전분, 탄산칼슘, 수크로스, 락토오스 및 젤라틴 등을 섞어 조제된다. 경구 투여를 위한 액상 제제로는 현탁제, 내용액제, 유제 및 시럽제 등을 들 수 있는데, 흔히 사용되는 단순 희석제인 물, 액체 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제 및 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결 건조 제제 및 좌제가 포함된다. 비수성용제와 현탁용제로는 프로필렌글리콜, 폴리에틸렌글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(Witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세롤 및 젤라틴 등이 사용될 수 있다. The composition according to this embodiment can be used as a pharmaceutical composition. Specifically, it is a skin irritant caused by acne, atopy, psoriasis, dermatitis and the like caused by inflammation reaction, skin aging caused by increase of active oxygen, wrinkles, and stain, freckles, blotch, It can be used as a pharmaceutical composition having a preventive or therapeutic effect on a skin pigmentation disease such as a pigment, a pigment, a drug-induced pigmentation, and an inflammation-induced pigmentation. The pharmaceutical composition can be administered at various times during clinical administration, including oral, transdermal, subcutaneous, intravenous, or muscular, and can be applied as an external preparation for skin. In addition, formulations can be prepared in various formulations. In the case of formulation, formulations can be prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants that are usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, and such solid preparations may contain at least one excipient such as starch, calcium carbonate, sucrose, lactose and gelatin, . Examples of liquid preparations for oral administration include suspensions, solutions, emulsions and syrups. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances and preservatives . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories. Examples of the non-aqueous solvent and the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Witepsol, macrogol, tween 61, cacao paper, laurin, glycerol and gelatin may be used as a base for suppositories.
또한, 목적하는 방법에 따라 경구 투여하거나 비경구 투여할 수 있으며, 비경구 투여시 피부 외용 또는 복강내주사, 직장내주사, 피하주사, 정맥주사, 근육내 주사 또는 흉부내 주사 주입방식을 선택하는 것이 바람직하다. 투여량은 환자의 체중, 연령, 성별, 건강상태, 식이, 투여시간, 투여방법, 배설율 및 질환의 중증도 등에 따라 그 범위가 다양하다. 투여량은 환자의 체중, 연령, 성별, 건강상태, 식이, 투여시간, 투여방법, 배설율 및 질환의 중증도에 따라 그 범위가 다양하며, 일일 투여량은 인진쑥 추출물 또는 이의 분획물의 양을 기준으로 0.0001 내지 100 mg/kg이고, 바람직하게는 0.001 내지 10 mg/kg이며, 하루 1 ~ 6 회 투여될 수 있다. 본 발명의 조성물은 단독으로, 또는 수술, 방사선 치료, 호르몬 치료, 화학 치료 및 생물학적 반응 조절제를 사용하는 방법들과 병용하여 사용할 수 있다.In addition, oral administration or parenteral administration may be carried out according to the intended method, and parenteral administration may be selected by external or intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection, . The dosage varies depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and disease severity. The dosage varies depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of disease, and the daily dosage is based on the amount of the extract or fraction thereof 0.0001 to 100 mg / kg, preferably 0.001 to 10 mg / kg, and can be administered 1 to 6 times a day. The composition of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers.
본 실시예에 의한 상기 조성물은 식품 조성물로 이용될 수 있다. 건강식품은 인진쑥 추출물 또는 이의 분획물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 건강식품의 종류에는 특별한 제한은 없으며, 예로는 육류, 소시지, 빵, 초콜릿, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알코올음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강식품을 모두 포함한다. 건강음료 조성물은 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토스, 슈크로스와 같은 디사카라이드, 및 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드, 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100 g 당 일반적으로 약 0.01 ~ 0.04 g, 바람직하게는 약 0.02 ~ 0.03 g 이다. 상기 외에 본 발명의 건강식품은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 밖에 천연 과일주스, 과일주스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 혼합하여 사용할 수 있다. 이러한 첨가제의 비율은 크게 중요하진 않지만 본 발명의 조성물 100 중량부당 0.01 ~ 0.1 중량부의 범위에서 선택되는 것이 일반적이다.The composition according to this embodiment can be used as a food composition. The health food can be used as it is or directly with other food or food ingredients, and can be suitably used according to conventional methods. There is no particular restriction on the type of health food. Examples include meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, soups, , An alcoholic beverage and a vitamin complex, and includes all the health foods in a conventional sense. The health beverage composition may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Such natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 g of the composition of the present invention. In addition to the above, the health food of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, , A carbonating agent used in carbonated drinks, and the like. It may also contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.
본 발명의 일 실시예에 따라 인진쑥 추출물로부터 분리된 류코딘(Leukodin)을 유효성분으로 함유하는 피부 미백용 조성물의 제조방법에 있어서, (1) 인진쑥(Artemisia Capillaris)에 물, 알코올 또는 이들의 혼합용매로 인진쑥 추출물을 제조하는 단계와, (2) 상기 인진쑥 추출물을 유기용매로 분획하여 인진쑥 분획물을 제조하는 단계를 포함할 수 있다. In the production method of the composition for skin whitening comprising the flow kodin (Leukodin) injinssuk separated from the extract according to one embodiment of the invention as an active ingredient, (1) injinssuk (Artemisia Capillaris ) with water, an alcohol or a mixed solvent thereof, and (2) fractionating the Artemisia sp. Extract with an organic solvent to produce Artemisia sp. Fractions.
단계 (1)은 인진쑥을 물, 알코올 또는 이들의 혼합물을 사용하여 인진쑥 추출물을 제조하는 하는 것으로서, 추출한 인진쑥 추출물을 여과 또는 감압 농축하고 건조 시키는 것일 수 있다. 상기 알코올로는 C1 내지 C2 저급 알코올을 이용하는 것이 바람직하며, 저급 알코올로는 에탄올 또는 메탄올을 이용하는 것이 바람직하다. 추출 방법으로는 진탕 추출, 속슬렛(Soxhlet) 추출 또는 환류 추출을 이용하는 것이 바람직하나 이에 한정되지 않는다. 상기 추출용매를 건조된 인진쑥 분량에 1 내지 5배 첨가하여 추출하는 것이 바람직하고, 1 내지 2배 첨가하여 추출하는 것이 더욱 바람직하며, 1.5배 첨가하는 것이 가장 바람직하다. 추출온도는 20 내지 100℃ 인 것이 바람직하고, 20 내지 40℃인 것이 더욱 바람직하고, 실온인 것이 가장 바람직하나, 이에 한정하지 않는다. 또한, 추출시간은 10 내지 48시간인 것이 바람직하며, 12 내지 36 시간인 것이 더욱 바람직하나, 이에 한정하지 않는다. 아울러, 추출 회수는 1 내지 5 회인 것이 바람직하며, 1 내지 2회 반복 추출하는 것이 더욱 바람직하고, 1회인 것이 가장 바람직하나, 이에 한정되는 것은 아니다. 인진쑥(Artemisia Capillaris)은 재배한 것 또는 시판되는 것 등 제한 없이 사용할 수 있다. 상기 인진쑥은 잎, 꽃, 뿌리, 줄기, 종자 등 모든 기관이 이용가능하고, 줄기 또는 잎인 것이 가장 바람직하나, 이에 한정되지 않는다. 추출물을 건조하는 것에 있어, 감압농축은 진공감압농축기 또는 진공회전증발기를 이용하는 것이 바람직하나 이에 한정하지 않으며, 이때 농축 농도는 당도 측정기로 측정하였을 때 약 2~10 브릭스(Brix)가 되도록 하는 것이 바람직하며, 3~6 브릭스가 되도록 하는 것이 더욱 바람직하나, 이에 한정되지 않는다. 또한, 건조는 감압건조, 진공건조, 비등건조, 분무건조 또는 동결건조하는 것이 바람직하나 이에 한정하지 않는다.Step (1) is to produce an extract of Artemisia japonica using water, alcohol or a mixture thereof, and the extracted Artemisia sp. Extract may be filtered or concentrated under reduced pressure and dried. As the alcohol, C 1 to C 2 lower alcohol is preferably used, and as the lower alcohol, ethanol or methanol is preferably used. As the extraction method, it is preferable to use shaking extraction, Soxhlet extraction or reflux extraction, but it is not limited thereto. The extraction solvent is preferably added 1 to 5 times, more preferably 1 to 2 times, more preferably 1.5 times, more preferably 1 to 5 times. The extraction temperature is preferably 20 to 100 ° C, more preferably 20 to 40 ° C, most preferably room temperature, but is not limited thereto. Further, the extraction time is preferably 10 to 48 hours, more preferably 12 to 36 hours, but is not limited thereto. The number of times of extraction is preferably 1 to 5 times, more preferably 1 to 2 times, more preferably 1 time, but is not limited thereto. Artemisia Capillaris ) may be used without restrictions, such as cultivated or marketed. It is most preferable that all organs such as leaves, flowers, roots, stems, and seeds are available, but it is not limited thereto. In the drying of the extract, it is preferable to use a vacuum decompression concentrator or a vacuum rotary evaporator for the concentration under reduced pressure, but the present invention is not limited thereto. The concentration of the concentrate is preferably about 2 to 10 Brix as measured by a sugar content meter More preferably 3 to 6 bricks, but it is not limited thereto. The drying is preferably performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying, but not always limited thereto.
단계(2)는 (1)단계에서 추출한 인진쑥 추출물을 유기용매로 분획 추출하여 인진쑥 분획 추출물을 제조하는 단계로서, 물, 알코올 또는 이들의 혼합 용액으로 추출된 인진쑥 추출물에서 류코딘(Leukodin)을 추출하기 위한 단계이다. 유기용매는 에틸아세테이트, 메탄올, 헥산, 또는 이의 혼합용액일 수 있고, 물과 에틸아세테이트 혼합용액, 메탄올과 헥산 혼합 용액인 것이 바람직하나, 이에 한정하지 않는다. 인진쑥 추출물을 물과 에틸아세테이트의 혼합 용액을 용매로 하여 분획하는 단계를 포함할 수 있고, 이를 통해 본 발명의 에틸아세테이트 분획물을 얻을 수 있다. 이때, 상기 물과 에틸아세테이트 혼합 용액 내 에틸아세테이트의 농도는 부피 농도로 40 내지 70%인 것이 바람직하고, 50 내지 60%인 것이 더욱 바람직하며, 50%인 것이 가장 바람직하나 이에 한정하지 않는다.Step (2) is a step of extracting the extract of Artemisia princeps, which is extracted in step (1), with an organic solvent to prepare Leucodyne extract. . The organic solvent may be ethyl acetate, methanol, hexane, or a mixed solution thereof. Preferably, the organic solvent is a mixed solution of water and ethyl acetate, or a mixed solution of methanol and hexane. And fractionating the extract of Artemisia mugwort with a mixture of water and ethyl acetate as a solvent, whereby the ethyl acetate fraction of the present invention can be obtained. At this time, the concentration of ethyl acetate in the mixed solution of water and ethyl acetate is preferably 40 to 70% by volume, more preferably 50 to 60%, most preferably 50% by volume.
에틸아세테이트 분획물을 메탄올과 헥산 혼합 용액을 용매로 하여 다시 분획하는 단계를 포함할 수 있고, 이를 통해 본 발명의 메탄올 분획물을 얻을 수 있다. 이때, 상기 메탄올과 헥산 혼합 용액 내 메탄올의 농도는 부피 농도로 70 내지 95%인 것이 바람직하고, 80 내지 90%인 것이 더욱 바람직하며, 90%인 것이 가장 바람직하나, 층 분리가 가능한 농도라면 모두 가능하다.And then fractionating the ethyl acetate fraction using a mixture of methanol and hexane as a solvent, whereby the methanol fraction of the present invention can be obtained. At this time, the concentration of methanol in the mixed solution of methanol and hexane is preferably 70 to 95% by volume, more preferably 80 to 90%, most preferably 90% It is possible.
또한, 상기 메탄올 분획물을 에틸아세테이트와 헥산 혼합 용액, 에틸아세테이트 또는 메틸알코올을 용매로 하여 분리하는 단계를 포함할 수 있고, 이를 통해 본 발명의 세부 분획물(Fr.1~Fr.8)을 얻을 수 있다. 이때, 상기 에틸아세테이트와 헥산 혼합 용액 내 에틸아세테이트의 농도는 10 내지 70%인 것이 바람직하고, 20 내지 50% 또는 55 내지 70%인 것이 더욱 바람직하며, 20 내지 50%, 또는 55 내지 66%인 것이 가장 바람직하나, 이에 한정하지 않는다. 분리는 순상 실리카겔(silica gel) 또는 역상 ODS(octadecylsiyl silica gel)를 이용할 수 있으나, 농도별 구배가 가능한 방법이라면 이에 한정하지 않는다.Further, the methanol fraction may be separated by using a mixed solvent of ethyl acetate and hexane, ethyl acetate or methyl alcohol as a solvent, and the fraction (Fr.1 to Fr.8) of the present invention can be obtained have. At this time, the concentration of ethyl acetate in the mixed solution of ethyl acetate and hexane is preferably 10 to 70%, more preferably 20 to 50% or 55 to 70%, more preferably 20 to 50%, or 55 to 66% Most preferably, but not exclusively. Separation can be performed using normal silica gel or reversed phase ODS (octadecylsiyl silica gel), but it is not limited to such a method as long as the concentration gradient is possible.
분획물의 제조 방법에서 추출 온도는 20 내지 100℃인 것이 바람직하고, 20 내지 40℃인 것이 더욱 바람직하며, 실온인 것이 가장 바람직하나 이에 한정하지 않는다. 또한, 추출 시간은 10 내지 48시간인 것이 바람직하며, 12 내지 36시간인 것이 더욱 바람직하나 이에 한정하지 않는다. 또한 분획 과정을 1 내지 5회, 바람직하게는 3회 반복하여 수득할 수 있고 분획 후 감압 농축하는 것이 바람직하나 이에 한정하지 않는다. In the method for preparing the fraction, the extraction temperature is preferably 20 to 100 ° C, more preferably 20 to 40 ° C, most preferably room temperature, but is not limited thereto. Further, the extraction time is preferably 10 to 48 hours, more preferably 12 to 36 hours, but is not limited thereto. Further, the fractionation process can be repeated 1 to 5 times, preferably 3 times, and it is preferable to fractionate and concentrate under reduced pressure, but it is not limited thereto.
또한, 본 발명의 일 실시예에 의한 제조방법에 있어서, 인진쑥 분획물에서 정제하는 단계를 더 포함할 수 있다. 이는 (2)단계에서 제조된 인진쑥 분획물(Fr.1~Fr.8)에 있어서, 류코딘(Leukodin)이 다량 포함될 수 있도록 분리 정제를 하기 위한 단계이며, 암모늄 아세테이트와 메탄올 혼합 용액을 용출액으로 하여 분리하는 단계를 포함할 수 있고, 이를 통해 본 발명의 인진쑥 추출물로부터 분리된 류코딘(Leukodin)을 얻을 수 있다. 본 실시예를 통해 인진쑥 추출물 또는 이의 분획물을 제조할 경우 활성 성분의 분산을 방지하고 더 많은 양의 활성 분획물을 얻을 수 있어, 적은 양으로도 피부 미백 활성을 가장 잘 나타낼 수 있는 추출물 또는 분획물을 효과적으로 얻을수 있다. In addition, the method according to an embodiment of the present invention may further include a step of purifying the fraction from Artemisia sp. This is a step for separating and purifying leucodyne (Leukodin) in the artificial squid fraction (Fr. 1 to Fr. 8) prepared in the step (2) by using a mixture solution of ammonium acetate and methanol as an eluent And isolating Leucodyne from the extract of Artemisia princeps of the present invention. When the extract of Artemisia princeps or its fractions is produced through the present example, it is possible to prevent the active ingredient from being dispersed and to obtain a larger amount of active fractions, so that the extract or fraction that can best exhibit skin whitening activity even in a small amount can be effectively Can be obtained.
인진쑥 추출물을 물과 에틸아세테이트 혼합 용액으로 추출하는 경우, 물 층에는 수용성 성분들이 포함되고, 에틸아세테이트 층에는 지용성 성분이 포함될 것이며, 이때 에틸아세테이트 층에서만 활성 효과가 나타났다는 것은 활성을 나타내는 성분이 대부분 지용성 성분이라는 것을 의미한다. 또한, 에틸아세테이트 층만을 분리하여, 메탄올과 헥산 혼합 용액으로 추출하는 경우, 헥산에는 지질 성분들이 포함되고, 메탄올 층에는 플라보노이드, 리그난 등의 극성이 다소 높은 성분이 포함될 것이며, 이때 메탄올 층에서만 활성 효과가 나타났다는 것은 활성을 나타내는 성분이 대부분 지용성 중에서도 극성이 다소 높은 성분이라는 것을 의미한다. When the extract of Artemisia officinalis is extracted with a mixture of water and ethyl acetate, the aqueous layer contains water-soluble components and the ethyl acetate layer contains a lipid-soluble component. At this time, only the ethyl acetate layer shows activity Soluble component. In addition, when only the ethyl acetate layer is separated and extracted with a mixed solution of methanol and hexane, hexane will contain lipid components, and the methanol layer will contain components with slightly higher polarity such as flavonoid and lignan, Means that the component exhibiting activity is a component having a somewhat higher polarity among fat-soluble components.
이에 반해, 통상적인 용매 분획법을 이용하였을 때는 극성에 따라 성분들이 다양한 용매에 세분화되어 포함될 것이기 때문에 활성 성분들이 분산됨으로써 본 발명과 같이, 활성성분이 대부분 극성을 띄는 경우에는 많은 양의 활성 성분을 얻는 데 불리할 것이다. 따라서 본 발명의 일 실시예에 따른 제조 방법은 인진쑥을 이용한 활성 성분의 분리시, 기존 방법의 한계를 극복하여 더 많은 양의 활성 성분을 손쉽게 얻을 수 있는 효과적인 방법이다. On the contrary, when a conventional solvent fractionation method is used, the active ingredients are dispersed in various solvents depending on the polarity, so that when the active ingredients are mostly polarized as in the present invention, a large amount of active ingredients It will be disadvantageous to obtain. Therefore, the production method according to one embodiment of the present invention is an effective method for easily obtaining a larger amount of active ingredient by overcoming the limitations of the existing method when the active ingredient is separated using Artemisia japonica.
이하 본 발명의 실시예에 대하여 첨부된 도면을 참조하여 그 구성 및 작용을 설명한다. 이 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지 않는 것은 당 업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will now be described in detail with reference to the accompanying drawings. It will be apparent to those skilled in the art that this embodiment is for illustrative purposes only and that the scope of the present invention is not construed as being limited by these embodiments.
실시예Example 1. 본 발명의 일 1. The present invention 실시예에In the embodiment 의한 인진쑥 에탄올 추출물의 제조 Preparation of Ethanol Extract of Artemisia capillaris
실험에 이용된 인진쑥은 전라남도 화순지역에서 채취하여 건조한 뒤 4℃에서 보관하여 사용한다. 100% 에탄올 (71.343 g)을 용매로 하여 실온에서 3일간 추출하고 여과지(Advantec No.2, Toyo, Japan)를 사용하여 감압 여과 한 후, 회전진공농축기(rotary vaccum evaporator)(EYELA, Japan)로 감압 농축하여 동결건조 한 것을 시료로 사용하였다(도 1). The mushroom collected from the Hwasun area in Jeollanam-do was dried and stored at 4 ℃. The mixture was extracted with 100% ethanol (71.343 g) as a solvent at room temperature for 3 days, filtered under reduced pressure using a filter paper (Advantec No.2, Toyo, Japan), and then transferred to a rotary vacuum evaporator (EYELA, Japan) Concentrated under reduced pressure, and lyophilized was used as a sample (Fig. 1).
실시예Example 2. 본 발명의 일 2. The present invention 실시예에In the embodiment 의한 인진쑥 Inosugi 분획물의Fraction 제조 Produce
실시예 1에 따라 제조된 에탄올 인진쑥 추출물의 극성별 용매 분획을 위해, 에탄올 인진쑥 추출물을 물과 에틸아세테이트(ethyl acetate)를 1:3로 혼합하여 용매분획 한 다음, 에틸아세테이트(ethyl acetate)층(38.56 g)을 다시 90% 메탄올(24.19 g)과 α-헥산(α-hexane)의 1:3으로 용매 분획하였다.제조된 인진쑥 90% 메탄올 분획물 5g을 에틸아세테이트20ml에 용해시킨 후, 실리카 겔 40(SIL-25㎍,YMC)과 MPLC를 실행하였다. 실험은 에틸아세테이트와 α-헥산(2:8 내지 10:0, 60분), 에틸아세테이트, 메틸알코올의 조건으로 전개시켜 8개의 분획(fraction, FR)으로 분리하였다(도 1).For the solvent fraction of the ethanolic Artemisia monocytogenes extract prepared according to Example 1, ethanol extract of Artemisia sp. Was mixed with water and ethyl acetate at a ratio of 1: 3, followed by solvent fractionation. Then, an ethyl acetate layer 38.56 g) was further subjected to solvent fractionation with 1: 3 of 90% methanol (24.19 g) and? -Hexane. 5 g of the prepared 90% methanol fraction of arsugi was dissolved in 20 ml of ethyl acetate, (SIL-25 占 퐂, YMC) and MPLC. The experiment was developed under the conditions of ethyl acetate and a-hexane (2: 8 to 10: 0, 60 minutes), ethyl acetate, methyl alcohol and separated into eight fractions (FR) (Fig. 1).
FR. 1-6
FR. 1-6
실시예Example 3. 인진쑥 추출물 내의 3. In the extract 류코딘Leucodyne (( LeukodinLeucodyne ) 분리 ) detach 정제 (refine ( preparation-preparation- HPLCHPLC , prep-, prep- HPLCHPLC ) 실시) Conducted
실시예 2에서 실시한 MPLC 분획물에서 미백활성이 류코딘(MPLC fraction 3)의 순도를 높이기 위한 분리 정제를 위해, 프렙-HPLC(prep-HPLC)용 YMC HPLC 컬럼을 이용하였다. 이때 시료 (MPLC fraction 3)는 200㎕를 투입하였다. 용출액은 2mM 암모늄 아세테이트(ammonium acetate)를 첨가한 40% 메탄올을 이용하였으며, 용출속도는 10㎖/min로 하였다. 검출기는 UV 검출기(254nm)를 사용하였다(도 3).A YMC HPLC column for prep-HPLC was used for separation purification to increase the purity of the whitening activity of the MPLC fraction 3 in the MPLC fraction performed in Example 2. [ At this time, 200 μl of sample (MPLC fraction 3) was added. The eluate was 40% methanol with 2 mM ammonium acetate, and the elution rate was 10 ml / min. The detector used was a UV detector (254 nm) (Figure 3).
실시예Example 4. 본 발명의 일 4. Work of the invention 실시예에In the embodiment 의한 정제된 인진쑥 Refined eggplant 분획물(FR.3)의The fraction (FR.3) HPLCHPLC 분석을 통한 Through analysis 류코딘Leucodyne (Leukodin) 분석.(Leukodin) analysis.
MPLC로 실시하여 분획된 인진쑥 분획물(FR.3)을 고성능액체크로마토그래피(Waters 2000 series, Japan)로 분석하였으며, 컬럼은 YMC C18 컬럼(5㎛, 250㎜×4.6㎜)을 사용하고, UV 검출기(254nm, 285nm, 366nm)를 사용하였다. 이동상은 물과 메탄올(methyl alcohol, MeOH)을 사용하였고 펌프 프로그램을 통해 기울기(gradient)를 설정하였으며, 유속은 1ml/min로 하였다. 이동상 용매의 조성과 기울기 조건은 하기 [표 3]에 보는 바와 같이 다양한 조건으로 설정하여 분석 실험을 수행하였다. 분석용으로 사용된 시료는 농도는 1㎎/㎖로 메탄올에 녹여 사용하였으며, 제조한 용액을 오토샘플러(autosampler)로 10㎕씩 주입하여 분석 실험을 실시하였다. (도 4) (FR.3) was analyzed by high performance liquid chromatography (Waters 2000 series, Japan). The column was a YMC C18 column (5 탆, 250 mm × 4.6 mm) (254 nm, 285 nm, 366 nm) was used. The mobile phase was water and methanol (MeOH), and the gradient was set through the pump program. The flow rate was 1 ml / min. The composition and slope conditions of the mobile phase solvent were set at various conditions as shown in Table 3 below, and the analysis experiments were performed. Samples used for analysis were dissolved in methanol at a concentration of 1 mg / ml, and 10 μl of the prepared solution was injected into an autosampler. (Figure 4)
(min)Time
(min)
실시예Example 5. 본 발명의 일 5. Work of the invention 실시예에In the embodiment 의한 인진쑥 추출물로부터 분리 정제된 Isolated from Artemisia sp. 류코딘(Leukodin)의Leukodin 핵자기공명(NMR) 분석 Nuclear magnetic resonance (NMR) analysis
분리된 화합물들의 구조를 구명하기 위해서 명지대학교 기기분석/정보지원센터의 400 MHz의 핵자기 분광분석기(Varian, 400-MR)를 사용하여 1H-NMR, 14C-NMR를 측정하였고, 분석용매로는 중클로로포름을 사용하였다(도 5 및 도 6). 1 H-NMR and 14 C-NMR were measured using a 400 MHz nuclear magnetic resonance spectrometer (Varian, 400-MR) from Myongji University Instrumental Analysis and Information Support Center to determine the structure of the separated compounds. Chloroform was used as the medium (Figs. 5 and 6).
실험예Experimental Example 1. One. 멜라닌세포(melanocyte)를Melanocytes (melanocytes) 이용한 피부 미백 효능 확인 Confirmed skin whitening efficacy
세포 배양은 B16F10 세포를 10% FBS, 1% pen/strep를 포함한 알파 MEM(alpha modification)으로 37℃, 5% CO2인큐베이터에서 배양하였다. 멜라닌 측정을 위해, B16F10 세포를 6개의 홈이 있는 플레이트의 각 홈(well)에 3.0×105cells/well을 깔아 37℃, 5% CO2에서 24시간 배양(pre-incubation)한 후, 알파 MEM 2ml로 교체하여 후보 소재의 추출물을 각 농도 별로 48시간 처리하였다. 처리 후 배지를 제거한 다음 PBS로 세척하고 0.05% 트립신-EDTA 처리하여 세포 펠렛을 회수하였다. 회수된 펠렛을 1.5ml 튜브로 옮긴 후, 3000rpm으로 5분 원심분리한 후 상등액을 제거하였다. 상등액이 제거된 펠렛을 1N 수산화나트륨(NaOH)을 50㎕를 넣고 2시간 동안 60℃에서 세포 내의 멜라닌을 용해 시켰다. 용해된 멜라닌을 96홈의 플레이트에 분주한 뒤, 405nm에서 흡광도를 측정한다(도 7 내지 도 9).B16F10 cells were cultured in a 5% CO 2 incubator at 37 ° C with alpha modification containing 10% FBS and 1% pen / strep. For melanin measurement, B16F10 cells were plated at 3.0 × 10 5 cells / well in each well of six grooved plates and preincubated at 37 ° C. and 5% CO 2 for 24 hours. And the extract of the candidate material was treated for 48 hours at each concentration by replacing with 2 ml of MEM. After the treatment, the medium was removed, washed with PBS, and treated with 0.05% trypsin-EDTA to recover the cell pellet. The recovered pellet was transferred to a 1.5 ml tube, centrifuged at 3000 rpm for 5 minutes, and the supernatant was removed. The pellet from which the supernatant was removed was added with 50 1 of 1 N sodium hydroxide (NaOH), and the melanin in the cells was dissolved at 60 캜 for 2 hours. The melted melanin is dispensed on a 96-well plate, and the absorbance is measured at 405 nm (Figs. 7 to 9).
이상, 본 발명내용의 특정한 부분을 상세히 기술하였는바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적인 기술은 단지 바람직한 실시태양일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의해 정의된다고 할 것이다.Having described specific portions of the present invention in detail, it will be apparent to those skilled in the art that this specific description is only a preferred embodiment and that the scope of the present invention is not limited thereby. It will be obvious. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.
Claims (9)
A skin whitening composition comprising Leukodin isolated from Artemisia sp. Extract as an active ingredient.
상기 조성물은 멜라닌 생성을 억제하는 것을 특징으로 하는 피부 미백용 조성물.
The method according to claim 1,
Wherein the composition inhibits melanin production.
상기 조성물은 화장료 조성물인 것을 특징으로 하는 피부 미백용 조성물.
The method according to claim 1,
Wherein the composition is a cosmetic composition.
상기 조성물은 비누 조성물인 것을 특징으로 하는 피부 미백용 조성물.
The method according to claim 1,
Wherein the composition is a soap composition.
상기 조성물은 약학적 조성물인 것을 특징으로 하는 피부 미백용 조성물.
The method according to claim 1,
Wherein the composition is a pharmaceutical composition.
상기 조성물은 식품 조성물인 것을 특징으로 하는 피부 미백용 조성물.
The method according to claim 1,
Wherein the composition is a food composition.
(1) 인진쑥(Artemisia Capillaris)에 물, 알코올 또는 이들의 혼합용매로 인진쑥 추출물을 제조하는 단계; 및
(2) 상기 (1)단계의 인진쑥 추출물을 유기용매로 분획하여 인진쑥 분획물을 제조하는 단계;
를 포함하는 인진쑥 추출물로부터 분리된 류코딘(Leukodin)을 유효성분으로 함유하는 피부 미백용 조성물의 제조방법.
A method for producing a skin whitening composition comprising Leukodin isolated from an Artemisia sp. Extract as an active ingredient,
(1) Artemisia Capillaris ) with water, an alcohol or a mixed solvent thereof; And
(2) fractionating the mugwort-mugwort extract of step (1) with an organic solvent to prepare a mugwort fraction;
(Leukodin) as an active ingredient isolated from an extract of Artemisia sp.
상기 인진쑥 분획물을 정제하는 단계를 더 포함하는 인진쑥 추출물로부터 분리된 류코딘(Leukodin)을 유효성분으로 함유하는 피부 미백용 조성물의 제조방법.
8. The method of claim 7,
The method for producing a skin whitening composition according to any one of claims 1 to 5,
상기 (2)단계의 유기용매는 에틸아세테이트, 메탄올, 헥산 또는 이들의 혼합 용액인 것을 특징으로 하는 피부 미백용 조성물의 제조방법.8. The method of claim 7,
Wherein the organic solvent in step (2) is ethyl acetate, methanol, hexane or a mixture thereof.
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KR20220028612A (en) * | 2020-08-31 | 2022-03-08 | 주식회사에이치엔비랩스 | Cosmetic composition antimicroboal, antioxidation, whitening or anti-inflammation comprising artemisia capillaris thunberg and olive extract |
KR20230067131A (en) * | 2021-11-09 | 2023-05-16 | 주식회사 씨앤비바이오 | A cosmetic composition containing a novel compound isolated from an Artemisia annua extract |
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KR20220028612A (en) * | 2020-08-31 | 2022-03-08 | 주식회사에이치엔비랩스 | Cosmetic composition antimicroboal, antioxidation, whitening or anti-inflammation comprising artemisia capillaris thunberg and olive extract |
KR102400179B1 (en) | 2020-08-31 | 2022-05-25 | 주식회사에이치엔비랩스 | Cosmetic composition antimicroboal, antioxidation, whitening or anti-inflammation comprising artemisia capillaris thunberg and olive extract |
KR20230067131A (en) * | 2021-11-09 | 2023-05-16 | 주식회사 씨앤비바이오 | A cosmetic composition containing a novel compound isolated from an Artemisia annua extract |
KR102576088B1 (en) | 2021-11-09 | 2023-09-08 | 주식회사 씨앤비바이오 | A cosmetic composition containing a novel compound isolated from an Artemisia annua extract |
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