KR102468538B1 - Skin improvement composition containing wild geranium extract with excellent antioxidant, whitening and anti-inflammatory effects - Google Patents
Skin improvement composition containing wild geranium extract with excellent antioxidant, whitening and anti-inflammatory effects Download PDFInfo
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- KR102468538B1 KR102468538B1 KR1020220043375A KR20220043375A KR102468538B1 KR 102468538 B1 KR102468538 B1 KR 102468538B1 KR 1020220043375 A KR1020220043375 A KR 1020220043375A KR 20220043375 A KR20220043375 A KR 20220043375A KR 102468538 B1 KR102468538 B1 KR 102468538B1
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- whitening
- extract
- wild geranium
- geranium extract
- wild
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- 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
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- A23V2200/00—Function of food ingredients
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- A23V2200/318—Foods, ingredients or supplements having a functional effect on health having an effect on skin health and hair or coat
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
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- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
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Abstract
Description
본 발명은 와일드제라늄 추출물이 함유되어 항산화, 미백 및 항염효능이 우수한 피부개선용 조성물에 관한 것으로, 더욱 상세하게는 피부적용시 안전하고 우수한 항산화 및 미백 효능을 나타낼 뿐만 아니라, 염증개선 효과를 나타내는 와일드제라늄 추출물을 유효성분으로 포함하는 피부개선용 조성물에 관한 것에 관한 것이다.The present invention relates to a composition for improving skin containing wild geranium extract and having excellent antioxidant, whitening and anti-inflammatory effects, and more particularly, when applied to the skin, it is safe and exhibits excellent antioxidant and whitening effects, as well as showing anti-inflammatory effects. It relates to a composition for skin improvement comprising a geranium extract as an active ingredient.
최근들어 성별과 나이를 불문하고 건강한 피부에 대한 관심이 높아지고 있으며, 이러한 사회적 트렌드에 따라 국내 기능성화장품 시장도 크게 성장하고 있다.Recently, interest in healthy skin has been increasing regardless of gender and age, and the domestic functional cosmetics market is growing significantly in accordance with this social trend.
식품의약품안전처(2021)에 따르면 화장품 수출액은 8.94조원으로 국내 총 수출의 1.40%를 차지하며, 연평균 성장률(2011년~2020년)은 29.19%로 나타났다. According to the Ministry of Food and Drug Safety (2021), cosmetics exports amounted to 8.94 trillion won, accounting for 1.40% of total domestic exports, and the average annual growth rate (2011-2020) was 29.19%.
특히, 친환경적인 천연유래 성분에 대한 소비자들의 관심이 높아지면서 다양한 브랜드에서 천연 유래 원료를 사용한 기능성 화장품을 출시하고 있다.In particular, as consumers' interest in eco-friendly, natural-derived ingredients increases, various brands are launching functional cosmetics using natural-derived ingredients.
기능성화장품 원료로서 천연물의 사용 가능성을 확인하는 대표적인 실험으로는 자유라디칼 소거능을 통한 항산화능 측정이 있다. 활성산소(reactive oxygen species, ROS)는 자유라디칼의 일종으로 생체 내 항산화 제거 기전에 의해 소거되고 있지만, 체내 활성산소의 생성 속도가 제거 속도보다 커지면 생체 조직을 공격할 뿐 아니라 정상 세포의 손상을 가져온다. 사람의 피부 및 세포에 손상을 일으키는 활성산소종은 강한 산화력으로 생체 내 세포를 구성하는 지질, 단백질, DNA 손상뿐만 아니라 진피층의 matrix metalloproteinase (MMP)-1 효소의 발현을 증가시켜 melanin 생성반응을 촉진 시킨다. 이를 통해 활성산소 소거가 멜라닌 색소 형성을 막을 수 있을 뿐 아니라 항산화 효과가 있는 물질이 멜라닌 색소 생성 억제에 효과를 나타낸다고 인식하게 되었다.As a representative experiment to confirm the possibility of using a natural product as a raw material for functional cosmetics, there is a measurement of antioxidant activity through free radical scavenging activity. Reactive oxygen species (ROS) is a type of free radical that is scavenged by the antioxidant removal mechanism in vivo. . Reactive oxygen species that cause damage to human skin and cells not only damage lipids, proteins, and DNA constituting cells in vivo with their strong oxidizing power, but also promote melanin production by increasing the expression of matrix metalloproteinase (MMP)-1 enzyme in the dermal layer. let it Through this, it was recognized that active oxygen scavenging can prevent melanin pigment formation, and substances with antioxidant effects are effective in inhibiting melanin pigment production.
멜라닌은 피부의 색을 결정하는 주요한 색소로서 표피의 기저층에 존재하는데, 합성기전으로는 tyrosinase의 촉매작용에 의한 타이로신이 hydroxyl와 결합되어 DOPA (3,4-dihydroxy phenylalanine)로 합성되는 효소 작용과, tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2) 및 microphthalmia-associated transcription factor (MITF)등 과 같은 단백질들이 관여하는 세포 내 신호전달 기전을 통한 합성이 있다. Arbutin, kojic acid, hydroquinone을 포함하는 많은 tyrosinase 저해제가 피부에 광범위하게 사용되고 있으나, 피부 자극 및 접촉성 피부염과 같은 부작용 사례들이 등장하면서 피부 친화적이고 안전한 천연물에서 이와 관련된 활성을 가진 소재를 찾으려는 각종 연구가 이어지고 있다.Melanin is the main pigment that determines the color of the skin and exists in the basal layer of the epidermis. The synthesis mechanism is the enzyme action that tyrosin is combined with hydroxyl by the catalytic action of tyrosinase to synthesize DOPA (3,4-dihydroxy phenylalanine), It is synthesized through intracellular signaling mechanisms involving proteins such as tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2) and microphthalmia-associated transcription factor (MITF). Many tyrosinase inhibitors, including arbutin, kojic acid, and hydroquinone, are widely used for skin, but as side effects such as skin irritation and contact dermatitis have emerged, various studies have been conducted to find materials with related activities from skin-friendly and safe natural products. is going on
MMP-1의 합성을 촉진하는 요소로 자외선, ROS뿐 아니라 염증성 cytokine인 interleukin (IL)-1과 IL-6 등이 있다. 생체 내에서 일어나는 염증 반응의 조절은 매우 복잡하며, 염증 과정 중에는 많은 양의 NO (nitric oxide)와 염증유도 사이토카인류 등이 생성되는 것으로 알려져 있다. NO는 superoxide 음이온 (O2 -)과 쉽게 반응하여 매우 반응성이 높고 독성이 강한 산화제인 peroxynitrite (ONOO-)를 생성할뿐 아니라, 과량 존재하면 인체에 유해한 영향을 미치게 되어 세포손상 및 염증 반응을 일으킨다. 즉, 염증 반응은 외인적 노화 요소가 피부 노화를 일으킬 수 있게 하는 중요한 반응이며, 함염 효과는 ROS의 생성 조절을 통해 피부 노화를 감소 시킬 수 있다.Factors that promote the synthesis of MMP-1 include ultraviolet rays, ROS, and inflammatory cytokines such as interleukin (IL)-1 and IL-6. It is known that the regulation of the inflammatory response occurring in vivo is very complex, and a large amount of NO (nitric oxide) and inflammation-inducing cytokines are produced during the inflammatory process. NO easily reacts with superoxide anion (O 2 - ) to produce peroxynitrite (ONOO - ), a highly reactive and highly toxic oxidizing agent. Excessive amounts of NO have harmful effects on the human body, causing cell damage and inflammatory reactions. . That is, the inflammatory response is an important reaction that allows extrinsic aging factors to cause skin aging, and the anti-inflammatory effect can reduce skin aging through the regulation of ROS production.
결국 여러 노화기전은 ROS의 생성과 연관이 있으며, 식물 유래 추출물에서 항산화 능을 나타내는 성분을 찾고자 많은 연구가 이루어지고 있다.After all, several aging mechanisms are related to the generation of ROS, and many studies are being conducted to find components that exhibit antioxidant activity in plant-derived extracts.
본 발명이 해결하고자 하는 과제는 피부적용시 안전하고 우수한 항산화 및 미백 효능을 나타낼 뿐만 아니라, 염증개선 효과를 나타내는 와일드제라늄 추출물을 유효성분으로 포함하는 피부개선용 조성물을 제공하는 것이다.The problem to be solved by the present invention is to provide a composition for skin improvement containing, as an active ingredient, a wild geranium extract that is safe and exhibits excellent antioxidant and whitening effects when applied to the skin, as well as an anti-inflammatory effect.
상기 과제를 해결하기 위하여, 본 발명은 와일드제라늄 추출물을 유효성분으로 포함하는 피부미백, 염증개선 및 항산화용 조성물(예를 들어, 화장료 조성물, 식품 조성물 등)을 제공한다.In order to solve the above problems, the present invention provides a composition for skin whitening, inflammation improvement and antioxidant (eg, cosmetic composition, food composition, etc.) containing wild geranium extract as an active ingredient.
본 발명의 바람직한 특징에 따르면, 상기 와일드제라늄 추출물은 정제수 100 중량부에 와일드제라늄 0.5 내지 1.5 중량부를 혼합하고 75 내지 85℃의 온도에서 30 내지 90분 동안 가열하여 제조되는 것으로 한다.According to a preferred feature of the present invention, the wild geranium extract is prepared by mixing 0.5 to 1.5 parts by weight of wild geranium with 100 parts by weight of purified water and heating at a temperature of 75 to 85 ° C. for 30 to 90 minutes.
그러나 이에 제한되는 것은 아니며 상기 열수의 온도는 예를들어 70 내지 130℃ 바람직하게는 80 내지 120℃, 보다 바람직하게는 90 내지 110℃일 수도 있다. 또한 추출방법은 가열추출기에서 1 내지 5시간, 바람직하게는 2 내지 4시간, 보다 바람직하게는 2시간 30분 내지 3시간 30분 동안 열수 추출하고, 이를 동결건조하여 수득하는 것일 수도 있으나 이에 제한되는 것은 아니다.However, it is not limited thereto, and the temperature of the hot water may be, for example, 70 to 130°C, preferably 80 to 120°C, and more preferably 90 to 110°C. In addition, the extraction method may be obtained by hot water extraction in a heating extractor for 1 to 5 hours, preferably 2 to 4 hours, more preferably 2 hours 30 minutes to 3 hours 30 minutes, and freeze-drying, but is limited thereto It is not.
또한, 본 발명의 목적은 상기 와일드제라늄 추출물이 함유되어 항산화, 미백 및 항염효능이 우수한 화장료용 조성물이 6.25 내지 500㎍/mL로 함유하는 것을 특징으로 하는 항산화, 미백 및 항염효능이 우수한 화장료를 제공함에 의해서도 달성될 수 있다.In addition, an object of the present invention is to provide a cosmetic composition with excellent antioxidant, whitening and anti-inflammatory effects, characterized in that the cosmetic composition containing the wild geranium extract and having excellent antioxidant, whitening and anti-inflammatory effects is contained at 6.25 to 500 μg / mL. can also be achieved by
그러나 상기 조성물은 화장료의 제형에 따라 또는 그것의 구체적인 적용 부위(얼굴, 손등, 발등, 목 등)등에 따라 바람직한 적용량이 달라지는 것이기 때문에, 상기 비율은 어떠한 측면으로든 본 발명의 범위를 제한하는 것으로 이해되어서는 안된다.However, since the preferred application amount of the composition varies depending on the formulation of the cosmetic or its specific application area (face, back of the hand, back of the foot, neck, etc.), it is understood that the ratio limits the scope of the present invention in any aspect. should not be
본 발명의 바람직한 특징에 따르면, 상기 항산화, 미백 및 항염효능이 우수한 화장료는 비누, 클렌징 폼, 클렌징 크림, 클렌징 워터, 목욕제, 스킨, 로숀, 크림, 에센스, 토너 및 바디클렌져로 이루어진 그룹에서 선택된 하나의 제형으로 제형화 될 수 있으나 이에 제한되는 것은 아니다.According to a preferred feature of the present invention, the cosmetic having excellent antioxidant, whitening and anti-inflammatory effects is one selected from the group consisting of soap, cleansing foam, cleansing cream, cleansing water, bath agent, toner, lotion, cream, essence, toner and body cleanser It may be formulated as a formulation of, but is not limited thereto.
또한, 본 발명의 조성물인 와일드제라늄 추출물은 피부와 같은 표적 영역으로의 운반을 위해 캡슐화 될 수 있으며, 캡슐화의 기술은 예를들어 피부에 성분을 전달하는 운반 비히클로서 사용될 수 있는 리포좀, 소낭 및 나노입자를 사용할 수 있으나 이에 제한되지 않는다.In addition, the wild geranium extract, the composition of the present invention, can be encapsulated for delivery to a target area such as the skin, and the technology of encapsulation can be used as a delivery vehicle to deliver the ingredients to the skin, for example, liposomes, vesicles, and nanoparticles. Particles may be used, but are not limited thereto.
또한, 본 발명의 목적은 와일드제라늄 추출물을 포함하는 이너뷰티(inner beauty)용 조성물을 제공함에 의해 달성될 수 있다.In addition, the object of the present invention can be achieved by providing a composition for inner beauty containing a wild geranium extract.
상기 이너뷰티 조성물은 보조제로 비타민, 무기질, 허브 또는 기타 식물, 아미노산, 효소 및 대사산물을 포함할 수 있으나 이에 제한되는 것은 아니다.The inner beauty composition may include vitamins, minerals, herbs or other plants, amino acids, enzymes and metabolites as adjuvants, but is not limited thereto.
또한, 본 발명의 목적은 와일드제라늄 추출물을 포함하는 건강기능식품 조성물을 제공함에 의해 달성될 수 있다.In addition, the object of the present invention can be achieved by providing a health functional food composition containing a wild geranium extract.
본 발명은 또 다른 하나의 양태로서, 와일드제라늄 추출물을 유효성분으로 함유하는 항산화, 미백, 또는 항염증용 의약외품 조성물을 제공한다.In another aspect, the present invention provides a quasi-drug composition for antioxidant, whitening, or anti-inflammatory containing wild geranium extract as an active ingredient.
본 발명에서, 와일드제라늄 추출물, 항산화, 미백 및 항염증은 전술한 바와 같다.In the present invention, wild geranium extract, antioxidant, whitening and anti-inflammatory are as described above.
본 발명에서 사용되는 용어 "의약외품"은 사람이나 동물의 질병을 진단, 치료, 개선, 경감, 처치 또는 예방할 목적으로 사용되는 물품들 중 의약품보다 작용이 경미한 물품들을 의미하는 것으로, 예를 들어 약사법에 따르면 의약외품이란 의약품의 용도로 사용되는 물품을 제외한 것으로, 사람ㆍ동물의 질병 치료나 예방에 쓰이는 제품, 인체에 대한 작용이 경미하거나 직접 작용하지 않는 제품 등이 포함된다.The term "quasi-drugs" used in the present invention refers to items with a milder effect than pharmaceuticals among items used for the purpose of diagnosing, treating, improving, mitigating, treating or preventing diseases of humans or animals, for example according to the Pharmaceutical Affairs Act. According to the definition, quasi-drugs are items excluding items used for pharmaceutical purposes, and include products used for the treatment or prevention of diseases in humans and animals, and products with minor or no direct action on the human body.
본 발명의 상기 의약외품 조성물은 바디 클렌저, 소독 청결제, 세정제, 주방용 세정제, 청소용 세정제, 치약, 가글제, 물티슈, 세제, 비누, 핸드 워시, 헤어세정제, 헤어 유연제, 가습기 충진제, 마스크, 연고제 및 필터 충진제로 이루어진 군에서 선택되는 제형으로 제조할 수 있으나, 이에 제한되는 것은 아니다.The quasi-drug composition of the present invention is a body cleanser, disinfectant cleanser, detergent, kitchen detergent, cleaning detergent, toothpaste, gargling agent, wet tissue, detergent, soap, hand wash, hair cleaner, hair softener, humidifier filler, mask, ointment and filter filler It can be prepared in a formulation selected from the group consisting of, but is not limited thereto.
본 발명의 조성물을 항산화, 미백, 또는 항염증 목적으로 의약외품에 포함시킬 경우, 상기 조성물을 그대로 포함하여 사용하거나 다른 의약외품 성분과 함께 사용할 수 있고, 통상적인 방법에 따라 적절하게 사용할 수 있다. 유효 성분의 혼합량은 사용 목적에 따라 적합하게 결정할 수 있으며, 본 발명에 의한 의약외품 조성물은 상기 와일드제라늄 추출물을 조성물 총 중량에 대하여 0.01~20 중량%로 함유할 수 있다.When the composition of the present invention is included in a quasi-drug for antioxidant, whitening, or anti-inflammatory purposes, the composition may be used as it is or used together with other quasi-drug ingredients, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of use, and the quasi-drug composition according to the present invention may contain the wild geranium extract in an amount of 0.01 to 20% by weight based on the total weight of the composition.
본 발명에 따른 와일드제라늄 추출물이 함유되어 항산화, 미백 및 항염효능이 우수한 피부개선용 조성물은 와일드제라늄 추출물이 함유되어 우수한 항산화 및 미백 효능을 나타낼 뿐만 아니라, 염증개선 효과를 나타내어 기능성 원료로 사용될 수 있는 탁월한 효과를 나타낸다.The composition for improving skin containing the wild geranium extract according to the present invention and having excellent antioxidant, whitening and anti-inflammatory effects contains wild geranium extract and exhibits excellent antioxidant and whitening effects, as well as an anti-inflammatory effect, which can be used as a functional raw material shows excellent effect.
도 1은 본 발명의 제조예 1을 통해 제조된 와일드제라늄 추출물의 DPPH 라디컬 소거능을 측정하여 나타낸 그래프이다.
도 2는 본 발명의 제조예 1을 통해 제조된 와일드제라늄 추출물의 ABTS 라디컬 소거능을 측정하여 나타낸 그래프이다.1 is a graph showing the DPPH radical scavenging ability of wild geranium extract prepared in Preparation Example 1 of the present invention.
Figure 2 is a graph showing the ABTS radical scavenging ability of the wild geranium extract prepared through Preparation Example 1 of the present invention.
이하에는, 본 발명의 바람직한 실시예와 각 성분의 물성을 상세하게 설명하되, 이는 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자가 발명을 용이하게 실시할 수 있을 정도로 상세하게 설명하기 위한 것이지, 이로 인해 본 발명의 기술적인 사상 및 범주가 한정되는 것을 의미하지는 않는다.Hereinafter, a preferred embodiment of the present invention and the physical properties of each component will be described in detail, but this is to be explained in detail so that a person having ordinary knowledge in the art to which the present invention belongs can easily practice the invention, This is not meant to limit the technical spirit and scope of the present invention.
본 발명에 따른 와일드제라늄 추출물이 함유되어 항산화, 미백 및 항염효능이 우수한 피부개선용 조성물은 와일드제라늄 추출물이 함유되며, 우수한 항산화, 미백 및 염증개선 효과를 나타낸다.The composition for skin improvement containing the wild geranium extract according to the present invention and having excellent antioxidant, whitening and anti-inflammatory effects contains the wild geranium extract and exhibits excellent antioxidant, whitening and anti-inflammatory effects.
상기 와일드제라늄추출물(Wild Geranium Extract ; Geranium Maculatum Extract)은 야생에서 자란 제라늄으로, 주요성분은 제라니올, 씨르토로넬롤, 리날올, 씨트로넬릴 포마테, 메톤 등이 있으며, 알려진 효능으로는 우수한 항산화효과를 나타낼 뿐만 아니라, 정서적으로 기분 상승효과, 우울불안행동 장애 및 스트레스 해소, 피부에는 수렴, 피지분비 정상화로 인한 염증완화, 건성습진, 비들, 여드름 제거, 주름살 완화 등에 효과를 나타낸다.The Wild Geranium Extract (Geranium Maculatum Extract) is wild-grown geranium, and its main ingredients include geraniol, sirtoronellol, linalol, citronellyl formate, metone, etc., and its known efficacy is excellent. In addition to showing antioxidant effects, it also shows effects such as emotionally enhancing mood, relieving depression and anxiety behavior disorder and stress, convergence on the skin, relieving inflammation due to normalization of sebum secretion, removing dry eczema, bead, acne, and relieving wrinkles.
또한, 상기의 와일드제라늄 추출물은 용매 추출이나 열수 추출을 통해 제조 될 수 있다. In addition, the wild geranium extract may be prepared through solvent extraction or hot water extraction.
본 발명에서 "추출물"이란 추출 방법, 추출 용매, 추출된 성분 및 형태를 불문하고 천연물의 유효성분을 뽑아내어 얻어진 물질을 의미한다. 용매 추출시, 상기 용매는 C1 내지 C4의 저급 알코올, 부틸렌글리콜, 프로필렌글리콜, 글리세린, 함수에탄올, 함수부틸렌글리콜, 함수프로필렌글리콜, 헥산 및 에틸아세테이트로 이루어진 그룹에서 선택된 하나 이상으로 이루어질 수 있으며, 상기 C1 내지 C4의 저급 알코올은 메탄올 또는 에탄올을 사용하는 것이 바람직하다.In the present invention, "extract" means a substance obtained by extracting an active ingredient of a natural product regardless of extraction method, extraction solvent, extracted component and form. In the solvent extraction, the solvent is composed of at least one selected from the group consisting of C 1 to C 4 lower alcohol, butylene glycol, propylene glycol, glycerin, hydrous ethanol, hydrous butylene glycol, hydrous propylene glycol, hexane and ethyl acetate It is preferable to use methanol or ethanol as the C 1 to C 4 lower alcohol.
상기 와일드제라늄 추출물은 열수 추출 하였으나 이에 제한 되는 것은 아니다. 이 외 추출방법으로는 열수 수출, 초음파 추출법, 여과법, 환류 추출법 등이 있으며, 여러 추출방법이 병용되어 수행 될 수도 있으나 단독으로 수행될 수도 있다.The wild geranium extract was extracted with hot water, but is not limited thereto. Other extraction methods include hot water export, ultrasonic extraction, filtration, and reflux extraction, and several extraction methods may be used in combination or may be performed alone.
또한, 상기 열수 추출은 정제수 100 중량부에 와일드제라늄 0.5 내지 1.5 중량부를 혼합하고 가열하여 이루어지는데, 상기 열수의 온도는 예를들어 70 내지 130℃, 바람직하게는 80 내지 120℃, 보다 바람직하게는 90 내지 110℃일 수 있으나 이에 제한되는 것은 아니다. 또한 추출방법은 가열추출기에서 1 내지 5시간, 바람직하게는 2 내지 4시간, 보다 바람직하게는 2시간 30분 내지 3시간 30분 동안 열수 추출하고, 이를 동결건조하여 수득하는 것일 수도 있으나 이에 제한되는 것은 아니다.In addition, the hot water extraction is performed by mixing 0.5 to 1.5 parts by weight of wild geranium with 100 parts by weight of purified water and heating. The temperature of the hot water is, for example, 70 to 130 ° C, preferably 80 to 120 ° C, more preferably It may be 90 to 110 ℃, but is not limited thereto. In addition, the extraction method may be obtained by hot water extraction in a heating extractor for 1 to 5 hours, preferably 2 to 4 hours, more preferably 2 hours 30 minutes to 3 hours 30 minutes, and freeze-drying, but is limited thereto It is not.
또한, 상기의 성분으로 이루어지는 와일드제라늄 추출물이 함유되어 항산화, 미백 및 항염효능이 우수한 피부개선용 조성물은 피부노화 억제 또는 개선, 피부 보호 효과, 피부 미백등의 효과를 포함 할 수 있다.In addition, the composition for improving skin containing wild geranium extract consisting of the above components and having excellent antioxidant, whitening and anti-inflammatory effects may include effects such as suppression or improvement of skin aging, skin protection effect, and skin whitening.
본 발명의 화장료 조성물은 통상적으로 화장료 조성물에서 추가할 수 있는 보조제를 하나 이상 더 포함 할 수 있다. 예를들어 화장품 분야에서 통상적으로 사용되는 임의의 다른 성분인 유기용매, 증점제, 보존제, 유화제, 계면활성제, 연화제, 겔화제, 항산화제, 안정화제, 금속이온봉제제, 방부제, 펩타이드, 자외선차단제, 방향제, 아로마, 비타민, 습윤화제, 염료 및 안료, 방향제, 배양액, 식물줄기세포, 혈행촉진제, 친수성 및 친유성계면활성제 등의 보조제를 하나 이상 더 포함할 수 있다.The cosmetic composition of the present invention may further include one or more adjuvants that may be added in conventional cosmetic compositions. For example, organic solvents, thickeners, preservatives, emulsifiers, surfactants, softeners, gelling agents, antioxidants, stabilizers, metal ion sealants, preservatives, peptides, sunscreens, One or more adjuvants such as fragrances, aromas, vitamins, wetting agents, dyes and pigments, fragrances, culture solutions, plant stem cells, blood circulation promoters, hydrophilic and lipophilic surfactants may be further included.
본 발명의 화장료 조성물에는 통상적으로 화장료 조성물에 배합되는 다른 성분을 배합할 수 있다. 예를 들면, 보습제, 에몰리엔트제, 유지성분, 계면활성제, 유기 및 무기 안료, 유기 분체, 자외선 흡수제, 살균제, 산화방지제, 식물 추출물, pH조절제, 알콜, 색소, 향료, 혈행 촉진제, 정제수, 점증제, 경화제, 유화제, 가용화제, 방부제, 항산화제, 분말 및 오일, 버터 등을 들 수 있다.The cosmetic composition of the present invention may be formulated with other ingredients that are normally formulated in cosmetic compositions. For example, moisturizers, emollients, oil and fat components, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, bactericides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, fragrances, blood circulation promoters, purified water, thickeners, hardeners, emulsifiers, solubilizers, preservatives, antioxidants, powders and oils, butter, and the like.
예를들어 상기 조성물은 글리세린, 부틸렌글라이콜, 세라마이드, 토코페롤, 알란토인, 판테놀, 아데노신, 나이아신아마이드 등으로 이루어진 군에서 선택된 하나 이상의성분을 추가로 포함할 수 있으나, 이에 제한되는 것은 아니고 식품의약품안전처에 등록된 화장품 원료 리스트 및 ICID(국제화장품원료집)에 등재되어 있는 성분 중에서 1종 이상을 더 포함할 수 있다.For example, the composition may further include at least one component selected from the group consisting of glycerin, butylene glycol, ceramide, tocopherol, allantoin, panthenol, adenosine, niacinamide, etc., but is not limited thereto. It may further include one or more of the ingredients listed in the list of cosmetic ingredients registered with the Ministry of Health and Welfare and ICID (International Cosmetic Ingredients).
또한, 상기 와일드제라늄 추출물은 6.25 내지 500㎍/mL의 함량으로 화장품 베이스에 함유되어 항산화, 미백 및 항염효능이 우수한 기능성 화장료를 제공할 수 있는데, 상기 화장품 베이스의 제형으로는 해당 분야에서 통상적으로 제조되는 어떠한 제형으로도 제조할 수 있으며, 예를들어 스킨, 로션, 에센스, 크림, 클렌징, 팩, 파우더, 스프레이 등으로 제형화 될 수 있으나 이에 제한되는 것은 아니다.In addition, the wild geranium extract can be contained in a cosmetic base in an amount of 6.25 to 500 μg / mL to provide a functional cosmetic with excellent antioxidant, whitening and anti-inflammatory effects. The cosmetic base formulation is conventionally prepared in the field. It can be prepared in any formulation that is, for example, skin, lotion, essence, cream, cleansing, pack, powder, spray, etc., but may be formulated, but is not limited thereto.
예를들어 피부에 사용되는 세럼, 앰플, 토너, 스킨, 스킨토너, 에센스, 로션, 아이크림외에도 자외선차단제로 사용하는 선크림, 선블록, 선스틱, 선젤, 선스프레이, 선오일 등으로 제형화 될 수 있으며, 세안제로 사용하는 클렌징오일, 클렌징 워터, 클렌징 크림, 클렌징 밀크, 클렌징 밤, 클렌징 티슈&패드, 클렌징 폼, 클렌징 파우더, 클렌징 젤, 클렌징 로션, 립앤아이 리무버, 클렌징 비누, 팩으로 사용하는 마스크 시트팩, 워시오프팩, 필오프팩, 슬리핑팩, 수분팩, 퓨리파잉팩, 아이팩, 코팩, 패드 등으로 제형화 될 수 있다.For example, in addition to serum, ampoule, toner, skin, skin toner, essence, lotion, and eye cream used for skin, it can be formulated into sunscreen used as sunscreen, sunblock, sunstick, sungel, sunspray, and sunoil. Cleansing Oil, Cleansing Water, Cleansing Cream, Cleansing Milk, Cleansing Balm, Cleansing Tissue & Pad, Cleansing Foam, Cleansing Powder, Cleansing Gel, Cleansing Lotion, Lip & Eye Remover, Cleansing Soap It can be formulated as a mask sheet pack, wash-off pack, peel-off pack, sleeping pack, moisture pack, purifying pack, eye pack, nose pack, pad, etc.
또한, 메이크업 용으로 사용되는 프라이머, 메이크업 베이스, 파운데이션, BB크림, 톤업크림, 컨실러, CC크림, 쿠션 , 틴티드 모이스처라이저, 파립우더 (압축파우더 팩트, 투웨이 케익), 치크, 블러셔, 하이라이터 ,쉐딩 등으로 제형화 될 수 있으며, 포인트 메이크업 용으로 사용되는 아이브로우, 아이섀도, 아이라이너, 마스카라, 립스틱, 립글로스, 립틴트, 라이너, 립밤 (립에센스,립크림,립필링,립패치등) 등으로 제형화 될 수 있다. In addition, primer used for makeup, makeup base, foundation, BB cream, tone-up cream, concealer, CC cream, cushion, tinted moisturizer, par lip powder (compressed powder pact, two-way cake), cheek, blusher, highlighter, shading It can be formulated into eyebrow, eye shadow, eye liner, mascara, lipstick, lip gloss, lip tint, liner, lip balm (lip essence, lip cream, lip filling, lip patch, etc.) used for point makeup. can be angry
이 외에도 바디케어 용으로 사용되는 비누, 입욕제 ,바디워시, 샤워 젤, 바디로션, 바디크림, 바디오일, 핸드크림 (로션), 풋크림, 데오드란트 등으로 제형화 될 수 있으며, 헤어케어 용으로 사용되는 샴푸, 린스, 트리트먼트, 헤어에센스, 헤어오일 등으로 제형화 될 수 있으나 이에 제한되는 것은 아니다.In addition, it can be formulated into soap, bath salt, body wash, shower gel, body lotion, body cream, body oil, hand cream (lotion), foot cream, and deodorant used for body care, and used for hair care. It may be formulated into shampoo, rinse, treatment, hair essence, hair oil, etc., but is not limited thereto.
이때, 상기 화장료에 함유된 항산화, 미백 및 항염효능이 우수한 화장료용 조성물의 함량이 6.25㎍/mL 미만이면, 화장료의 항산화, 미백 및 염증개선 효능이 저하되며, 상기 항산화 및 미백효능이 우수한 화장료용 조성물의 함량이 500㎍/mL를 초과하게 되면 화장료의 항산화, 미백 및 염증개선 효능은 크게 향상되지 않으면서 피부 독성이 증가하여 피부트러블을 유발할 수 있고, 제조비용을 지나치게 증가시키기 때문에 바람직하지 못하다.At this time, if the content of the cosmetic composition with excellent antioxidant, whitening and anti-inflammatory effects contained in the cosmetic is less than 6.25 μg / mL, the antioxidant, whitening and anti-inflammatory efficacy of the cosmetic is lowered, and the cosmetic composition with excellent antioxidant and whitening is effective. When the content of the composition exceeds 500 μg / mL, the antioxidant, whitening, and anti-inflammatory effects of the cosmetic are not greatly improved, but skin toxicity increases, causing skin trouble, and manufacturing cost is undesirably increased.
또한, 본 발명은 와일드제라늄 추출물을 포함하는 이너뷰티(inner beauty)용 조성물을 제공한다.In addition, the present invention provides a composition for inner beauty comprising a wild geranium extract.
본 명세서에서 이너뷰티(inner beauty)란 먹는 화장품을 의미하는 것으로, 화장품을 바르는 것을 통해 피부 표면을 일시적으로 좋게 만드는 것뿐 아니라 식품 등을 통해 피부 내부에서부터 건강한 피부를 가꾸는 것을 의미한다.In this specification, inner beauty refers to cosmetics to be eaten, and means to make the skin surface temporarily better by applying cosmetics, as well as to maintain healthy skin from the inside of the skin through food.
이하에서는, 본 발명에 따른 와일드제라늄 추출물이 함유되어 항산화, 미백 및 항염효능이 우수한 피부개선용 조성물의 제조방법 및 그 제조방법으로 제조된 피부개선용 조성물의 물성을 실시예를 들어 설명하기로 한다.Hereinafter, a method for preparing a composition for improving skin containing wild geranium extract having excellent antioxidant, whitening and anti-inflammatory effects according to the present invention and the physical properties of the composition for improving skin prepared by the manufacturing method will be described with examples. .
<제조예 1> <Production Example 1>
정제수 100 중량부에 와일드제라늄 1 중량부를 혼합하고 80℃의 온도에서 60분 동안 가열한 후에 고형분을 걸러내어 와일드제라늄 추출물을 제조하였다.Wild geranium extract was prepared by mixing 1 part by weight of wild geranium with 100 parts by weight of purified water, heating at a temperature of 80 ° C. for 60 minutes, and filtering out the solid content.
상기 제조예 1을 통해 제조된 와일드제라늄 추출물의 물성을 측정하여 아래 도 1 내지 2 및 표 1 내지 5에 나타내었다.The physical properties of the wild geranium extract prepared in Preparation Example 1 were measured and are shown in Figures 1 to 2 and Tables 1 to 5 below.
{이때, 상기 와일드제라늄 추출물의 항산화능 실험 및 세포 실험을 위해 동결건조를 통해 추출물의 건조중량을 측정하였으며, 이를 증류수로 재용해시킨 뒤, syringe filter (PVDF, 0.2 μm)를 통해 균 및 불용물을 제거하였다.{At this time, the dry weight of the extract was measured through freeze-drying for the antioxidant activity test and cell test of the wild geranium extract, and after re-dissolving it in distilled water, bacteria and insoluble matter were removed through a syringe filter (PVDF, 0.2 μm). has been removed.
또한, 상기 와일드제라늄 추출물의 물성은 DPPH 라디컬 소거능, ABTS 라디컬 소거능, FRAP, 폴리페놀 함량, 플라보노이드 함량, 세포 독성, 미백 활성 및 항염을 측정하는 방법을 이용하였다.In addition, the physical properties of the wild geranium extract were measured using DPPH radical scavenging activity, ABTS radical scavenging activity, FRAP, polyphenol content, flavonoid content, cytotoxicity, whitening activity, and anti-inflammatory properties.
- DPPH 라디컬 소거능 측정- Measurement of DPPH radical scavenging ability
DPPH assay 측정은 라디컬 활성에 따라 500-550 nm 파장의 흡광도가 변화하는 인공적 라디컬 분자인 2,2-diphenyl-1-picrylhydrazyl을 이용하여 라디컬의 소거능을 측정하는 방법으로, DPPH 용액은 70% ethanol을 용매로 DPPH를 1%(w/v)가 되도록 녹인 뒤 filter paper를 이용해 불용된 DPPH를 제거하였다. 그 후 이 용액의 520 nm 흡광도가 1.00이 되도록 70% ethanol로 다시 희석하여 사용하였다. 500 μg/mL를 기준으로 2배씩 희석하여 만들어진 다양한 농도의 와일드제라늄 추출물 1.0 mL와 DPPH 용액 1.0 mL를 혼합 후 30 분간 반응시켰다. 그 후 520 nm에서의 흡광도를 측정하였다. 이때 추출물 및 환원된 DPPH 용액의 흡광도를 측정하여 실험결과에서 제외했다. 실험결과는 ascorbic acid의 EC50을 측정 후 비교하여 항산화능을 측정하였다.DPPH assay measurement is a method of measuring the radical scavenging ability by using 2,2-diphenyl-1-picrylhydrazyl, an artificial radical molecule whose absorbance of 500-550 nm wavelength changes according to radical activity. After dissolving DPPH to 1% (w/v) in % ethanol as a solvent, undissolved DPPH was removed using filter paper. After that, the solution was diluted with 70% ethanol again so that the absorbance at 520 nm was 1.00. After mixing 1.0 mL of wild geranium extract and 1.0 mL of DPPH solution at various concentrations prepared by diluting 2-fold based on 500 μg/mL, the mixture was reacted for 30 minutes. After that, the absorbance at 520 nm was measured. At this time, the absorbance of the extract and the reduced DPPH solution was measured and excluded from the experimental results. The experimental results measured the antioxidant capacity by comparing the EC 50 of ascorbic acid.
- ABTS 라디컬 소거능 측정- Measurement of ABTS radical scavenging ability
ABTS assay 측정은 ABTS의 라디컬 활성이 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)의 산화도에 따라 700-750 nm 파장의 흡광도가 변화하는 성질을 이용하는 것으로, ABTS 용액은 증류수를 용매로 7 mM ABTS를 녹인 뒤 2.45 mM potassium persulfate를 녹여 12 시간 동안 ABTS의 발색이 일어나도록 하였다. 발색이 끝난 ABTS 용액의 740 nm 흡광도가 0.7이 되도록 희석하여 사용하였다. 500 μg/mL를 기준으로 2배씩 희석하여 만들어진 다양한 농도의 와일드제라늄 추출물 1.0 mL와 ABTS 용액 1.0 mL를 혼합 후 30 분간 반응시켰다. 그 후 740 nm에서의 흡광도를 측정하였다. 이때 추출물 및 환원된 ABTS 용액의 흡광도를 측정하여 실험결과에서 제외했다. 실험결과는 ascorbic acid의 EC50을 측정 후 비교하여 항산화능을 측정하였다.ABTS assay measurement uses the property that the absorbance of 700-750 nm wavelength changes according to the degree of oxidation of 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) in the radical activity of ABTS. After dissolving 7 mM ABTS in distilled water as a solvent, 2.45 mM potassium persulfate was dissolved to allow ABTS to develop color for 12 hours. The ABTS solution after color development was diluted so that the absorbance at 740 nm was 0.7. After mixing 1.0 mL of wild geranium extract of various concentrations prepared by diluting 2-fold based on 500 μg/mL and 1.0 mL of ABTS solution, they were reacted for 30 minutes. After that, the absorbance at 740 nm was measured. At this time, the absorbance of the extract and the reduced ABTS solution was measured and excluded from the experimental results. The experimental results measured the antioxidant capacity by comparing the EC 50 of ascorbic acid.
- FRAP 측정- FRAP measurement
와일드제라늄 추출물의 환원력을 측정하기 위해 산화된 철 이온을 이용하였으며, 철 이온은 산화 정도에 따라 2가 이온과 3가 이온으로 나누어지며 이를 이용해 환원력 측정하였다. 실험에는 0.5 mg/mL 농도로 희석한 와일드제라늄 추출물을 사용하였다. 와일드제라늄 추출물 2.5 mL과 0.2 M 인산염 완충용액 (pH 6.6) 2.5 mL를 혼합하여 일정한 pH가 나타나도록 한 뒤, 10% 페리시안화 칼륨용액 2.5 ml를 주입하여 50℃에서 20분 동안 반응시켰다. 반응이 끝난 시료는 10% trichloroacetic acid 2.5 ml를 넣은 뒤 원심분리(3000 × G, 15 min)하여 침전물을 제거하였다. 침전물이 제거된 용액 1 mL와 0.1% ferric chloride 용액 0.2 mL를 혼합시켰다. 그 후 700 nm에서 흡광도를 측정하여 환원력을 평가하였다. FRAP은 ascorbic acid를 기준으로 standard curve를 작성하여 측정하였다.Oxidized iron ions were used to measure the reducing power of wild geranium extract, and iron ions were divided into divalent and trivalent ions according to the degree of oxidation, and the reducing power was measured using these. Wild geranium extract diluted to a concentration of 0.5 mg/mL was used in the experiment. After mixing 2.5 mL of wild geranium extract and 2.5 mL of 0.2 M phosphate buffer solution (pH 6.6) to obtain a constant pH, 2.5 mL of 10% potassium ferricyanide solution was injected and reacted at 50 °C for 20 minutes. After the reaction, the sample was centrifuged (3000 × G, 15 min) after adding 2.5 ml of 10% trichloroacetic acid to remove the precipitate. 1 mL of the solution from which the precipitate was removed and 0.2 mL of a 0.1% ferric chloride solution were mixed. After that, the absorbance was measured at 700 nm to evaluate the reducing power. FRAP was measured by preparing a standard curve based on ascorbic acid.
- 폴리페놀 함량 측정- Measurement of polyphenol content
폴리페놀의 경우 환원력을 가지고 있으며, 환원력 측정에 이용되는 Folin-Cioc
- 플라보노이드 함량 측정- Measurement of flavonoid content
플라보노이드의 경우 알루미늄 이온과 반응하여 510 nm의 흡광도를 내는 것을 이용하여 그 양을 측정하였는데, 플라보노이드 함량 측정은 플라보노이드와 알루미늄 이온의 결합에 따른 발색 차이를 통해 측정하였다. 와일드제라늄 추출물 1.0 mL에 NaNO2 (5%, w/v) 0.3 mL를 주입한 뒤 5 분 동안 실온에서 방치하였다. 그 후 AlCl3 (2%, w/v) 0.5 mL를 주입한 뒤 6 분 간 방치하였다. 1 M NaOH 0.5 mL를 주입하여 중화시킨 뒤 510 nm에서의 흡광도를 측정하였다. 플라보노이드 농도는 quercetin을 이용한 standard curve를 이용해 환산하였다.In the case of flavonoids, the amount was measured using those that react with aluminum ions to produce absorbance at 510 nm. After injecting 0.3 mL of NaNO 2 (5%, w/v) into 1.0 mL of wild geranium extract, the mixture was left at room temperature for 5 minutes. After that, 0.5 mL of AlCl 3 (2%, w/v) was injected and left for 6 minutes. After neutralization by injecting 0.5 mL of 1 M NaOH, absorbance at 510 nm was measured. The flavonoid concentration was converted using a standard curve using quercetin.
- 세포 독성 측정- Measurement of cytotoxicity
NADH 등의 세포 내의 환원성 물질에 의해 MTT가 결정화 되는 것을 이용하여 세포의 활성 및 생존율을 측정하였다. 세포 독성 실험에는 B16F10 cell와 RAW 264.7 cell을 사용하였다. 세포 배양 배지로는 DMEM broth (Dulbecco's Modified Eagle Medium, GE healthcare, USA)를 사용하였으며 FBS (fetal bovine serum, Sigma, USA)를 각각 5%, 10%를 첨가하여 제조하였다. 항생물질로 Penicillin-Streptomycin (100X) (Sigma, USA)을 사용하였다. 추출물은 DMSO에 용해시켜 처리하였다. 세포 독성의 측정에는 MTT assay를 실시하였다. 전배양된 세포를 96 well plate에 well 당 5.0×104 cell을 주입하여 24 시간 배양하였다. 배양이 끝난 후 상층액을 제거한 뒤 와일드제라늄 추출물을 100 μg/mL를 기준으로 다양한 농도로 처리한 뒤 72 시간 동안 배양하였다. 2차 배양 후 상층액을 제거하고 MTT 용액(5 mg/mL)를 가해준 뒤 온도 37℃, CO2 농도 5% 의 환경에서 MTT를 결정화시켰다. 각 well에 생성된 결정이 제거되지 않도록 상층액을 제거한 뒤 결정을 DMSO로 녹여 540 nm 파장에서의 흡광도를 측정하였다.Cell activity and viability were measured using the crystallization of MTT by intracellular reducing substances such as NADH. B16F10 cells and RAW 264.7 cells were used for cytotoxicity experiments. DMEM broth (Dulbecco's Modified Eagle Medium, GE healthcare, USA) was used as the cell culture medium, and FBS (fetal bovine serum, Sigma, USA) was added at 5% and 10%, respectively. Penicillin-Streptomycin (100X) (Sigma, USA) was used as an antibiotic. The extract was treated by dissolving in DMSO. MTT assay was performed for the measurement of cytotoxicity. The pre-cultured cells were cultured for 24 hours by injecting 5.0×10 4 cells per well into a 96 well plate. After the culture was completed, the supernatant was removed, and the wild geranium extract was treated with various concentrations based on 100 μg/mL and cultured for 72 hours. After the secondary culture, the supernatant was removed, MTT solution (5 mg/mL) was added, and MTT was crystallized in an environment of 37°C and 5% CO 2 concentration. After removing the supernatant so as not to remove the crystals generated in each well, the crystals were dissolved in DMSO and the absorbance at a wavelength of 540 nm was measured.
- 미백 활성 측정- Measurement of whitening activity
미백 활성 측정은 멜라닌을 측정하는 방법을 이용하였으며, 피부색을 결정하는 요소인 멜라닌 생성을 유도하는 호르몬을 처리한 뒤 와일드제라늄 추출물을 처리하여 멜라닌 감소양을 측정 하였다.To measure the whitening activity, a method of measuring melanin was used, and after treatment with a hormone that induces melanin production, which is a factor determining skin color, wild geranium extract was treated to measure the amount of melanin reduction.
B16F10 cell을 사용하여 멜라닌 생성량 측정 실험을 진행하였다. 세포 배양 배지로는 DMEM broth를 사용하였으며, FBS 5%를 첨가하여 제조하였다. 항생물질로는 Penicillin-Streptomycin (100X)을 사용하였다. 추출물은 DMSO에 용해시켜 처리하였다. 멜라닌 측정을 위해 B16F10을 96 well plate에 각 well 당 5.0×104cell 씩 seeding하여 1일간 37℃, 5% CO2 조건에서 세포를 배양하였다. 배양이 끝난 후 상층액을 제거한 뒤 와일드제라늄 추출물을 100 μg/mL를 기준으로 다양한 농도로 처리하고 α-MSH의 농도가 0.1 μg/mL가 되도록 처리하여 72 시간 동안 배양하였다. 2차 배양 후 10% DMSO가 포함된 1N NaOH 0.1 μL를 처리 후 1시간동안 방치하여 멜라닌을 용해시킨 후 405nm 흡광도를 측정하였다. An experiment was conducted to measure the amount of melanin produced using B16F10 cells. DMEM broth was used as a cell culture medium, and it was prepared by adding 5% FBS. Penicillin-Streptomycin (100X) was used as an antibiotic. The extract was treated by dissolving in DMSO. To measure melanin, B16F10 was seeded in 5.0×10 4 cells per well in a 96 well plate, and the cells were cultured at 37°C and 5% CO2 for 1 day. After the culture was completed, the supernatant was removed, and the wild geranium extract was treated at various concentrations based on 100 μg/mL, and the α-MSH concentration was treated to be 0.1 μg/mL, followed by incubation for 72 hours. After the secondary culture, 0.1 μL of 1N NaOH containing 10% DMSO was treated and allowed to stand for 1 hour to dissolve melanin, and then the absorbance at 405 nm was measured.
-항염- anti-inflammatory
염증 전달 물질 중 하나인 NO에 대한 농도를 측정하여 염증 억제 효과를 확인하였다. 세포 염증 실험에는 RAW 264.7 cell을 사용하였다. 세포 배양 배지로는 DMEM broth를 사용하였으며 FBS 10%를 첨가하여 제조하였다. 항생물질로는 Penicillin-Streptomycin (100X)을 사용하였다. 추출물은 DMSO에 용해시켜 처리하였다. NO 측정을 위해 RAW 264.7을 96 well plate에 각 well 당 5.0×104cell 씩 seeding하여 1일간 37℃, 5% CO2 조건에서 세포를 배양하였다. 배양이 끝난 후 상층액을 제거한 뒤 와일드제라늄 추출물을 100 μg/mL를 기준으로 다양한 농도로 처리하고 LPS (lipopolysaccharide, Sigma, USA)의 농도가 1 μg/mL가 되도록 처리하여 48 시간 동안 배양하였다. 2차 배양 후 상층액을 회수하여 상층액 100 μL와 griess reagent 100μL를 혼합하여 와일드제라늄 추출물의 NO 농도를 측정하였다.}The anti-inflammatory effect was confirmed by measuring the concentration of NO, one of the inflammatory transmitters. RAW 264.7 cells were used for cell inflammation experiments. As a cell culture medium, DMEM broth was used and prepared by adding 10% FBS. Penicillin-Streptomycin (100X) was used as an antibiotic. The extract was treated by dissolving in DMSO. For NO measurement, RAW 264.7 was seeded in 5.0×10 4 cells per well in a 96 well plate, and the cells were cultured at 37° C. and 5% CO2 for 1 day. After the culture was finished, the supernatant was removed, and the wild geranium extract was treated at various concentrations based on 100 μg/mL, and the concentration of LPS (lipopolysaccharide, Sigma, USA) was 1 μg/mL, and cultured for 48 hours. After the secondary culture, the supernatant was recovered, and the NO concentration of wild geranium extract was measured by mixing 100 μL of the supernatant with 100 μL of griess reagent.}
상기 제조예 1을 통해 제조된 와일드제라늄 추출물의 DPPH 라디컬 소거능을 측정하여 아래 도 1에 나타내었다.The DPPH radical scavenging ability of the wild geranium extract prepared in Preparation Example 1 was measured and shown in FIG. 1 below.
실험에 앞서 비교대상으로 사용된 ascorbic acid의 EC50 수치를 측정하였다. 각 농도는 3.125-50 μg/mL 농도 범위로 반응시켜 ascorbic acid의 EC50 수치는 17.44 μg/mL임을 확인했다.Prior to the experiment, the EC 50 value of ascorbic acid used as a comparison was measured. Each concentration was reacted in the concentration range of 3.125-50 μg/mL, and it was confirmed that the EC 50 value of ascorbic acid was 17.44 μg/mL.
한편 와일드제라늄 추출물의 경우 31.25-500 μg/mL로 희석하여 실험을 진행하였다. 실험 결과 31.25 μg/mL 농도에서 27.1±2.04%, 62.5 μg/mL 농도에서 47.94±1.87%, 125 μg/mL 농도에서 77.42±1.87%, 250 μg/mL 농도에서 95.01±0.19%, 500 μg/mL 농도에서 96.91±1.73%의 DPPH 라디칼 소거능이 나타났다. 이를 바탕으로 EC50을 결과는 65.59 μg/mL였다.Meanwhile, in the case of wild geranium extract, the experiment was conducted by diluting it to 31.25-500 μg/mL. Experimental results: 27.1±2.04% at 31.25 μg/mL concentration, 47.94±1.87% at 62.5 μg/mL concentration, 77.42±1.87% at 125 μg/mL concentration, 95.01±0.19% at 250 μg/mL concentration, 500 μg/mL The DPPH radical scavenging ability of 96.91±1.73% was shown at this concentration. Based on this, the EC 50 was 65.59 μg/mL.
한편 EC50을 통한 효과는 ascorbic acid 는 17.44 μg/mL, 본 추출물은 65.59 μg/mL로 나타나 ascorbic acid와 비교 시 약 25.59%의 항산화능을 보였다.On the other hand, the effect through EC 50 was 17.44 μg/mL for ascorbic acid and 65.59 μg/mL for this extract, showing about 25.59% of antioxidant activity compared to ascorbic acid.
이를 유사한 방법으로 추출한 민들레(Taraxacum officinal)와 비교하였을 때 500 μg/mL 농도에서 추출 부위에 따라 66.20-87.66%의 DPPH 라디칼 소거능이 나타났다. 와일드제라늄은 동일 농도에서 96.91±1.73%의 DPPH 라디칼 소거능이 나타나, 와일드제라늄은 민들레에 비해 약 1.11- 1.46배 강한 항산화능을 나타내는 것을 알 수 있다.When compared with dandelion ( Taraxacum officinal ) extracted in a similar way, DPPH radical scavenging ability of 66.20-87.66% was shown depending on the extraction site at a concentration of 500 μg / mL. Wild geranium showed DPPH radical scavenging activity of 96.91±1.73% at the same concentration, and wild geranium showed about 1.11-1.46 times stronger antioxidant activity than dandelion.
또한, 제조예 1을 통해 제조된 와일드제라늄 추출물의 ABTS 라디컬 소거능을 측정하여 아래 도 2에 나타내었다.In addition, the ABTS radical scavenging ability of the wild geranium extract prepared in Preparation Example 1 was measured and shown in FIG. 2 below.
실험에 앞서 비교대상으로 사용된 ascorbic acid의 EC50 수치를 측정하였다. 각 농도는 3.125-50 μg/mL 농도 범위로 반응시켜 ascorbic acid의 EC50 수치는 9.11 μg/mL임을 확인했다.Prior to the experiment, the EC 50 value of ascorbic acid used as a comparison was measured. Each concentration was reacted in the concentration range of 3.125-50 μg/mL, and it was confirmed that the EC 50 value of ascorbic acid was 9.11 μg/mL.
한편 와일드제라늄 추출물의 경우 31.25-500 μg/mL로 희석하여 실험을 진행하였다. 실험 결과 31.25 μg/mL 에서 29.36±1.35%, 62.5 μg/mL 에서 57.65±3.69%, 125 μg/mL 에서 82.31±1.69%, 250 μg/mL 에서 97.02±2.17%, 0.32 μg/mL 에서 98.48±1.87%의 ABTS 라디칼 소거능이 나타났다. EC50 계산 결과 54.05 μg/mL란 결과를 얻었다. Meanwhile, in the case of wild geranium extract, the experiment was conducted by diluting it to 31.25-500 μg/mL. Experimental results 29.36±1.35% at 31.25 μg/mL, 57.65±3.69% at 62.5 μg/mL, 82.31±1.69% at 125 μg/mL, 97.02±2.17% at 250 μg/mL, 98.48±1.87 at 0.32 μg/mL % ABTS radical scavenging activity was shown. As a result of EC 50 calculation, 54.05 μg/mL was obtained.
한편 EC50을 통한 효과 비교시 ascorbic acid 는 9.11 μg/mL, 와일드제라늄 추출물은 54.05 μg/mL로 나타나 ascorbic acid와 비교 시 약 16.86%의 항산화능을 보였다.Meanwhile, when comparing the effect through EC 50 , ascorbic acid was 9.11 μg/mL and wild geranium extract was 54.05 μg/mL, showing about 16.86% of antioxidant activity compared to ascorbic acid.
이를 대나무잎 열수추출물과 비교한 결과 품종에 따라 133.30-199.14 μg/mL의 EC50을 나타내었으며, 와일드제라늄의 EC50은 54.05 μg/mL로 대나무잎의 2.47-3.68배 강한 항산화능을 나타내었다.As a result of comparing this with the bamboo leaf hot water extract, the EC 50 was 133.30-199.14 μg/mL depending on the variety, and the EC 50 of wild geranium was 54.05 μg/mL, showing 2.47-3.68 times stronger antioxidant activity than that of bamboo leaf.
또한, 상기 제조예 1을 통해 제조된 와일드제라늄 추출물의 FRAP를 측정한 결과 와일드제라늄 추출물의 FRAP는 0.097±0.004로 나타났으며, ascorbic acid의 standard curve를 이용하여 와일드제라늄 추출물의 1 mg의 FRAP은 ascorbic acid 0.229±0.009 mg의 FRAP과 같음을 알 수 있었다.In addition, as a result of measuring FRAP of the wild geranium extract prepared through Preparation Example 1, the FRAP of the wild geranium extract was 0.097 ± 0.004, and 1 mg of FRAP of the wild geranium extract was obtained using the standard curve of ascorbic acid. It was found to be the same as FRAP of ascorbic acid 0.229±0.009 mg.
또한, 상기 제조예 1을 통해 제조된 와일드제라늄 추출물의 폴리페놀 및 플라보노이드 함량을 측정하여 아래 표 1에 나타내었다.In addition, the polyphenol and flavonoid contents of the wild geranium extract prepared in Preparation Example 1 were measured and shown in Table 1 below.
<표 1><Table 1>
상기 표 1에 나타낸 것처럼, 폴리페놀의 경우 32.989±1.610 mg/g으로 나타났으며, 플라보노이드의 경우 11.098±0.261 mg/g으로 나타났다.As shown in Table 1, polyphenols were found to be 32.989 ± 1.610 mg / g, and flavonoids were found to be 11.098 ± 0.261 mg / g.
또한, 상기 제조예 1을 통해 제조된 와일드제라늄 추출물의 세포독성을 측정하여 아래 표 2 내지 3에 나타내었다.In addition, the cytotoxicity of the wild geranium extract prepared in Preparation Example 1 was measured and shown in Tables 2 to 3 below.
{단, 와일드제라늄 추출물의 세포에 미치는 독성을 평가하기 위해서 MTT assay를 이용한 세포 생존률을 측정하였다. 와일드제라늄 추출물은 배지에서의 최종농도가 6.25-100 μg/mL가 되도록 배합하여 배양을 진행하였다.}{However, in order to evaluate the toxicity of wild geranium extract to cells, cell viability was measured using MTT assay. Wild geranium extract was cultured by blending so that the final concentration in the medium was 6.25-100 μg / mL.}
<표 2> Cytotoxicity of Geranium Maculatum Extract on B16F10<Table 2> Cytotoxicity of Geranium Maculatum Extract on B16F10
(* p<.05, ** p<.01, *** p<.001)(*p<.05, **p<.01, ***p<.001)
상기 표 2에 나타낸 것처럼, 6.25 μg/mL 농도에서 3.35±0.46%, 12.5 μg/mL 농도에서 5.03±0.33%, 25 μg/mL 농도에서 7.42±1.03%, 50 μg/mL 농도에서 11.23±0.93%, 100 μg/mL 농도에서 14.71±0.71%의 세포 생존률이 나타났다. , 50 μg/mL 농도에서 11.23±0.93%, 100 μg/mL 농도에서 14.71±0.71%의 세포 생존률이 나타났다.As shown in Table 2 above, 3.35 ± 0.46% at 6.25 μg / mL concentration, 5.03 ± 0.33% at 12.5 μg / mL concentration, 7.42 ± 1.03% at 25 μg / mL concentration, 11.23 ± 0.93% at 50 μg / mL concentration , a cell viability of 14.71±0.71% was observed at a concentration of 100 μg/mL. , cell viability of 11.23±0.93% at 50 μg/mL concentration and 14.71±0.71% at 100 μg/mL concentration.
<표 3> Cytotoxicity of Geranium Maculatum Extract on RAW 264.7<Table 3> Cytotoxicity of Geranium Maculatum Extract on RAW 264.7
(* p<.05, ** p<.01)(*p<.05, **p<.01)
상기 표 3에 나타낸 것처럼, 6.25 μg/mL 농도에서 1.58±1.32%, 12.5 μg/mL 농도에서 2.03±1.29%, 25 μg/mL 농도에서 3.42±0.91%, 50 μg/mL 농도에서 7.63±0.79%, 100 μg/mL 농도에서 10.25±0.47%의 세포 생존률이 나타났다. As shown in Table 3 above, 1.58 ± 1.32% at 6.25 μg / mL concentration, 2.03 ± 1.29% at 12.5 μg / mL concentration, 3.42 ± 0.91% at 25 μg / mL concentration, 7.63 ± 0.79% at 50 μg / mL concentration , a cell viability of 10.25±0.47% was observed at a concentration of 100 μg/mL.
ISO 10993-5 및 식품의약품안전처 고시 제2014-115호의 기준에서 20% 이상의 세포 독성을 나타내었을 때 독성이 있는 것으로 간주하며, 이러한 기준으로는 모든 실험 농도에서 세포독성이 나타나지 않은 것을 알 수 있다.In accordance with ISO 10993-5 and Ministry of Food and Drug Safety Notice No. 2014-115, it is considered to be toxic when cytotoxicity of 20% or more is exhibited, and it can be seen that cytotoxicity does not occur at all experimental concentrations based on these standards. .
또한, 상기 제조예 1을 통해 제조된 와일드제라늄 추출물의 미백활성을 측정하여 아래 표 4에 나타내었다.In addition, the whitening activity of the wild geranium extract prepared in Preparation Example 1 was measured and shown in Table 4 below.
{단, 와일드제라늄 추출물이 멜라닌 생성에 미치는 영향을 평가하기 위해서 B16F10의 멜라닌 생성량을 측정하되, 추출물은 배지에서 최종농도가 6.25-100 μg/mL가 되도록 배합하여 배양을 진행하였다.}{However, in order to evaluate the effect of the wild geranium extract on melanin production, the amount of melanin production of B16F10 was measured, but the culture was carried out by mixing the extract so that the final concentration in the medium was 6.25-100 μg / mL.}
<표 4> Melanin production rate of B16F10 with Geranium Maculatum Extract<Table 4> Melanin production rate of B16F10 with Geranium Maculatum Extract
(** p<.01)(**p<.01)
상기 표 4에 나타낸 것처럼, 6.25 μg/mL 농도에서 21.71±3.74%, 12.5 μg/mL 농도에서 31.58±3.13%, 25 μg/mL 농도에서 39.31±2.13%, 50 μg/mL 농도에서 39.47±1.01%, 100 μg/mL 농도에서 40.62±2.07%의 미백활성이 나타났다.As shown in Table 4 above, 21.71 ± 3.74% at 6.25 μg / mL concentration, 31.58 ± 3.13% at 12.5 μg / mL concentration, 39.31 ± 2.13% at 25 μg / mL concentration, 39.47 ± 1.01% at 50 μg / mL concentration , 40.62±2.07% of whitening activity was shown at 100 μg/mL concentration.
상황버섯 추출물과 미백활성을 비교할 경우 열수 추출물에서 양쪽 최대 30% 이상의 미백효과를 내었으나 저농도 조건인 25 μg/mL에서의 결과를 비교했을때, 상황버섯의 경우 21.46%의 미백효과를 보인데 비해, 와일드제라늄 추출물의 경우 39.31%의 미백효과를 보여 약 1.8배 더 높은 미백효과를 나타내었다.When comparing Phellinus linteus extract and whitening activity, the hot water extract showed a whitening effect of up to 30% or more, but when comparing the results under low concentration conditions of 25 μg/mL, Phellinus linteus showed a whitening effect of 21.46%, whereas , Wild geranium extract showed a whitening effect of 39.31%, which was about 1.8 times higher.
또한, 상기 제조예 1을 통해 제조된 와일드제라늄 추출물의 항염성능을 측정하여 아래 표 5에 나타내었다.In addition, the anti-inflammatory performance of the wild geranium extract prepared in Preparation Example 1 was measured and shown in Table 5 below.
{단, 와일드제라늄 추출물이 염증 전달물질에 미치는 영향을 평가하기 위해서 RAW 264.7의 NO 생성량을 측정하였다. 추출물은 배지에서 최종농도가 6.25-100 μg/mL가 되도록 배합하여 배양을 진행하였다.}{However, in order to evaluate the effect of wild geranium extract on inflammatory mediators, the NO production amount of RAW 264.7 was measured. The extract was cultured by mixing the final concentration in the medium to be 6.25-100 μg / mL.}
<표 5><Table 5>
(* p<.05, ** p<.01)(*p<.05, **p<.01)
상기 표 5에 나타낸 것처럼, 염증과 관련된 지표인 NO의 생성률을 계산한 결과이다. 12.5 μg/mL 에서 6.79±3.81%, 25 μg/mL 에서 11.07±2.87%, 50 μg/mL 에서 16.07±2.67%, 100 μg/mL 에서 27.86±2.82%의 염증 억제율이 나타났다. As shown in Table 5 above, it is the result of calculating the production rate of NO, which is an index related to inflammation. Inflammation inhibition was 6.79±3.81% at 12.5 μg/mL, 11.07±2.87% at 25 μg/mL, 16.07±2.67% at 50 μg/mL, and 27.86±2.82% at 100 μg/mL.
민들레의 열수 추출물의 항염효과와 비교할 경우 250 μg/mL에서 7.5%의 감소를 보인데 비해 본 추출물은 100 μg/mL 에서 27.86±2.82%의 염증 억제율을 나타내 낮은 농도에서도 더 높은 항염활성을 나타내었다.Compared to the anti-inflammatory effect of the hot water extract of dandelion, a decrease of 7.5% at 250 μg/mL, whereas this extract showed a 27.86±2.82% anti-inflammatory rate at 100 μg/mL, showing higher anti-inflammatory activity even at lower concentrations. .
따라서, 본 발명에 따른 와일드제라늄 추출물이 함유되어 항산화, 미백 및 항염효능이 우수한 피부개선용 조성물은 와일드제라늄 추출물이 함유되어 우수한 항산화 및 미백 효능을 나타낼 뿐만 아니라, 염증개선 효과를 나타내어 기능성 원료로 사용될 수 있다.Therefore, the composition for improving skin containing the wild geranium extract according to the present invention and having excellent antioxidant, whitening and anti-inflammatory effects contains the wild geranium extract and exhibits excellent antioxidant and whitening effects, and also exhibits an anti-inflammatory effect, so it can be used as a functional raw material. can
Claims (6)
상기 피부개선용 조성물은 와일드제라늄 추출물로 이루어지고,
상기 와일드제라늄 추출물은 정제수 100 중량부에 와일드제라늄 1 중량부를 혼합하고 80℃의 온도에서 60분 동안 가열하여 제조되는 것을 특징으로 하는 항산화, 미백 및 항염효능을 나타내는 화장료.
The composition for skin improvement is contained at 31.25 to 100 μg/mL,
The composition for improving the skin is made of wild geranium extract,
The wild geranium extract is a cosmetic that exhibits antioxidant, whitening and anti-inflammatory effects, characterized in that prepared by mixing 1 part by weight of wild geranium with 100 parts by weight of purified water and heating at a temperature of 80 ° C. for 60 minutes.
상기 항산화, 미백 및 항염효능을 나타내는 화장료는 비누, 클렌징 폼, 클렌징 크림, 클렌징 워터, 목욕제, 스킨, 로숀, 크림, 에센스, 토너 및 바디클렌져로 이루어진 그룹에서 선택된 하나의 제형으로 제조되는 것을 특징으로 하는 항산화, 미백 및 항염효능을 나타내는 화장료.The method of claim 3,
Cosmetics exhibiting antioxidant, whitening and anti-inflammatory effects are prepared in one formulation selected from the group consisting of soap, cleansing foam, cleansing cream, cleansing water, bath agent, toner, lotion, cream, essence, toner and body cleanser Characterized in that Cosmetics that exhibit antioxidant, whitening and anti-inflammatory effects.
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| JP2012051906A (en) * | 2011-10-05 | 2012-03-15 | Ichimaru Pharcos Co Ltd | Trypsin inhibitor |
| KR101126984B1 (en) | 2011-12-14 | 2012-03-21 | 제니코스(주) | Cosmetic composition comprising extract of taxodium distichum and viburnum sargentii for sterile for whitening and antioxdiant |
| KR20160087113A (en) * | 2015-01-13 | 2016-07-21 | 정다운 | Cosmetics containing flowers |
| JP2018512409A (en) * | 2015-03-16 | 2018-05-17 | ザ ブーツ カンパニー ピーエルシー | Topical cosmetic compositions for free radicals |
| KR20190121091A (en) * | 2018-04-17 | 2019-10-25 | 제주대학교 산학협력단 | Whitening composition containing extract, fractions or compound derived from Raphanus sativus L. var niger |
| KR102343342B1 (en) | 2020-12-16 | 2021-12-24 | 유창우 | Antioxidant cosmetic composition containing vitamin c and houttuynia cordata extract |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JP2012051906A (en) * | 2011-10-05 | 2012-03-15 | Ichimaru Pharcos Co Ltd | Trypsin inhibitor |
| KR101126984B1 (en) | 2011-12-14 | 2012-03-21 | 제니코스(주) | Cosmetic composition comprising extract of taxodium distichum and viburnum sargentii for sterile for whitening and antioxdiant |
| KR20160087113A (en) * | 2015-01-13 | 2016-07-21 | 정다운 | Cosmetics containing flowers |
| JP2018512409A (en) * | 2015-03-16 | 2018-05-17 | ザ ブーツ カンパニー ピーエルシー | Topical cosmetic compositions for free radicals |
| KR20190121091A (en) * | 2018-04-17 | 2019-10-25 | 제주대학교 산학협력단 | Whitening composition containing extract, fractions or compound derived from Raphanus sativus L. var niger |
| KR102343342B1 (en) | 2020-12-16 | 2021-12-24 | 유창우 | Antioxidant cosmetic composition containing vitamin c and houttuynia cordata extract |
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