KR102472370B1 - Cosmetic composition with excellent antioxidant and whitening effects - Google Patents
Cosmetic composition with excellent antioxidant and whitening effects Download PDFInfo
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- KR102472370B1 KR102472370B1 KR1020220032981A KR20220032981A KR102472370B1 KR 102472370 B1 KR102472370 B1 KR 102472370B1 KR 1020220032981 A KR1020220032981 A KR 1020220032981A KR 20220032981 A KR20220032981 A KR 20220032981A KR 102472370 B1 KR102472370 B1 KR 102472370B1
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- GVJHHUAWPYXKBD-IEOSBIPESA-N α-tocopherol Chemical compound OC1=C(C)C(C)=C2O[C@@](CCC[C@H](C)CCC[C@H](C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-IEOSBIPESA-N 0.000 description 1
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- 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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Abstract
Description
본 발명은 항산화 및 미백효능이 우수한 화장료용 조성물에 관한 것으로, 더욱 상세하게는 동물성 원료를 사용하지 않으면서도 우수한 항산화 및 미백 효능을 나타내어 기능성 비건 화장료의 원료로 사용될 수 있는 항산화 및 미백효능이 우수한 화장료용 조성물에 관한 것이다.The present invention relates to a cosmetic composition with excellent antioxidant and whitening effects, and more particularly, a cosmetic composition with excellent antioxidant and whitening effects that can be used as a raw material for functional vegan cosmetics by showing excellent antioxidant and whitening effects without using animal raw materials. It is about a composition for
최근 사회적인 요구에 부응하듯 화장품 제조 과정 중 동물실험 및 동물성 원료를 사용하지 않는 비건 뷰티(vegan beauty)가 증가하고 있다. 비거니즘(veganism)은 인류의 건강뿐 아니라 지구상의 고통받는 동물을 줄이기 위한 동물해방, 환경오염, 온난화 현상으로 인한 기후 변화 등 생태 환경을 파괴하는 것을 멈추기 위한 행동이다1). 실제로 무분별한 동물성 원료 사용은 생태계를 파괴시키는 요인으로 문제시되었고, 이미 유럽연합(EU)은 화장품 지침 전문을 통해 화장품의 원료 또는 원료의 배합과정 및 완성된 화장품의 안전성 평가를 시행해야 할 경우 86/609/EEC (동물실험 보호를 규정한 지침)를 적용해야 한다고 규정하였다.Recently, vegan beauty, which does not use animal testing and animal-derived ingredients in the cosmetic manufacturing process, is increasing as if to meet social demands. Veganism is an action to stop destroying the ecological environment, such as animal liberation, environmental pollution, and climate change due to global warming to reduce human suffering as well as human health 1) . In fact, indiscriminate use of animal raw materials has been problematic as a factor that destroys the ecosystem, and the European Union (EU) has already issued 86/609 when it is necessary to evaluate the safety of cosmetic raw materials or the mixing process of raw materials and finished cosmetics through the full text of the cosmetics directive. /EEC (Guidelines regulating the protection of animal experiments) should be applied.
이러한 움직임은 세계 각국으로 확대되어 국내에서도 화장품법 15조 2항에 따라 동물실험을 실시한 화장품의 유통·판매를 법적으로 금지시켰으며, 현재 국내 기업들도 뷰티 시장 변화의 흐름을 반영하여 동물성 원료 대신 자연 유래 친환경 성분만을 사용하여 화장품을 연구·개발·생산하는 데 동참하고 있다. 아모레퍼시픽의 경우 2008년부터 화장품의 원료뿐 아니라 완제품에 대한 동물실험을 금지하기 시작하여 2013년부터는 협력업체에도 같은 방침을 적용하였다. 또한 LG생활건강도 2012년부터 전 제품에 대한 동물실험을 중단하였으며, 세포배양 독성평가법 또는 면역세포 배양 평가법 등으로 이를 대체하고 있다. 이처럼 점차적으로 비건뷰티 제품을 개발하는 활동이 활발해지며 소비자들 또한 친환경적이면서 좋은 성분이 함유되어 있는 비건원료 화장품에 대한 관심과 수요가 크게 증가하고 있는 것이 현실이다.This movement has expanded to countries around the world, and distribution and sales of cosmetics that have been tested on animals have been legally banned in Korea in accordance with Article 15, Paragraph 2 of the Cosmetics Act. We are participating in research, development, and production of cosmetics using only derived eco-friendly ingredients. In the case of AMOREPACIFIC, animal testing for finished products as well as raw materials for cosmetics began to be banned in 2008, and the same policy was applied to partner companies from 2013. In addition, LG Household & Health Care also stopped animal testing for all products from 2012, and is replacing it with a cell culture toxicity evaluation method or an immune cell culture evaluation method. As such, the activity of developing vegan beauty products is gradually becoming active, and consumers are also experiencing a significant increase in interest and demand for vegan raw cosmetic products that are environmentally friendly and contain good ingredients.
2019년 4월 Industry Reports(인더스트리 리포트)에 따르면 전 세계 비건 원료를 사용한 화장품 시장 규모를 2018년에서 2023년까지의 기간 동안 7.1%의 연평균 성장률을 기록할 것으로 예상하였으며, 또 다른 글로벌 시장조사기관인 Statista(스태티스타)도 역시 2020년 150억 달러(약 17조원)에서 2025년에는 약 41% 증가한 200억 달러(약 24조원)에 이를 것으로 전망한 바 있다. 또한 미국의 Data Bridge(데이터 브릿지) 시장조사에서도 2020년 4월 기준으로 2027년까지 236억 6000만 달러의 규모 가치를 예상하고, 6.25%의 성장률을 전망하여 비건 뷰티는 필수가 되고 있다.According to Industry Reports (Industry Report) in April 2019, the global cosmetics market size using vegan ingredients was expected to record an average annual growth rate of 7.1% during the period from 2018 to 2023, and another global market research institute, Statista (Statista) also predicted that it will reach 20 billion dollars (about 24 trillion won), an increase of about 41% from 15 billion dollars (about 17 trillion won) in 2020. In addition, in the US Data Bridge market research, as of April 2020, the scale value is expected to be $ 23.66 billion by 2027, and a growth rate of 6.25% is expected, making vegan beauty a must.
전 세계적으로 비건 화장품 시장이 크게 성장하고 있음에도 아직 비건 화장품 원료에 대한 연구는 미비하다. 그러나 화학성분의 인체에 대한 부작용이 대두됨에 따라 국내 원료업체들도 비건 소재에 대한 연구가 활발히 진행되고 있으며, 연구개발을 통해 일부 천연 원료들의 비건인증을 획득하고 있다. 이에 본 연구에서는 비건 인증 원료의 기능성 화장품 원료로서 사용 가능성을 검증하고자 항산화능 측정 및 멜라닌 측정을 통해 미백 효과를 확인하고자 한다.Although the vegan cosmetics market is growing significantly around the world, research on vegan cosmetics raw materials is still insufficient. However, as the side effects of chemical ingredients on the human body are soybean, domestic raw material companies are actively conducting research on vegan materials, and some natural raw materials are acquiring vegan certification through research and development. Therefore, in this study, in order to verify the possibility of using vegan-certified raw materials as functional cosmetics raw materials, the whitening effect was confirmed by measuring antioxidant capacity and melanin.
한편, 항산화능이란 다른 분자와 반응하기 쉬운 free radical을 제거하는 성질이다. 대표적인 free radical로는 활성산소(reactive oxygen species, ROS)가 있으며 이들은 세포의 노화에 소거능을 통해 측정한다. 우리의 인체는 대사작용 및 면역작용에 의해 ROS가 생성되며, 생성된 ROS를 조절하기 위해 superoxide dismutase, glutathione peroxidase, catalase 등과 같은 항산화 효소를 통한 효소적 항산화 기전과 ascorbic acid, tocopherol 등 비효소적 항산화 기전을 통해 조절하는 시스템이 이용된다. 그러나 여러 가지 요인들에 의해 산화촉진물질(prooxidant)과 산화억제물질(antioxidant)에 불균형이 생기면 산화 스트레스(oxidative stress)가 유발되고, 항산화 방어막이 파괴되어 피부 질환 및 노화를 초래하게 된다11). 현재 산화적 스트레스에 의한 세포 구성 성분의 산화적 손상을 지연시키거나 억제하는 물질인 항산화 물질을 다양한 천연소재에서 찾고자 하는 노력이 많이 시도되고 있으며, 이들이 가지는 항산화 특성을 이용한 기능성 소재의 개발에 대한 연구가 꾸준히 진행되고 있다.On the other hand, antioxidant activity is the property of removing free radicals that are easy to react with other molecules. Representative free radicals include reactive oxygen species (ROS), which are measured through scavenging activity in cell aging. ROS are generated by metabolism and immunity in the human body, and to control the generated ROS, enzymatic antioxidant mechanisms through antioxidant enzymes such as superoxide dismutase, glutathione peroxidase, and catalase, and non-enzymatic antioxidants such as ascorbic acid and tocopherol A system that regulates through a mechanism is used. However, when there is an imbalance between prooxidants and antioxidants due to various factors, oxidative stress is induced, and the antioxidant barrier is destroyed, resulting in skin diseases and aging [11] . Currently, many efforts are being made to find antioxidants, which are substances that delay or inhibit oxidative damage of cell components caused by oxidative stress, from various natural materials, and research on the development of functional materials using their antioxidant properties. is progressing steadily.
또한, 건강한 피부와 관련하여 항산화와 함께 중요도가 높아지는 분야 중 하나가 미백이다. 1999년에 화장품법이 약사법으로부터 독립 제정되고, 2000년 7월 1일 그 시행이 확정된 이후 안정적이고 효과적인 미백 소재를 발견하고자 하는 연구가 현재까지 활발히 진행되고 있기 때문이다. 그 결과 멜라닌 생성과 제어 메커니즘 연구는 진보를 이루었고, 이러한 연구 성과를 기초로 약용식물이나 천연물에서 이와 관련된 활성을 가진 소재를 찾으려는 각종 연구가 이어지고 있다. 그 중에서도 피부에 부작용이 나타나는 합성물질보다 비교적 안전성이 뛰어난 천연 추출물을 이용한 기능성 화장품 개발이 요구되고 있다. 천연 추출물에서 미백 효과를 나타내는 대표적인 성분은 polyphenol이 있다. Polyphenol은 생물 전반에서 발견되는 물질로서 일반적으로 항산화능을 나타내며, 항산화능은 멜라닌 생성의 주요 기전 중 하나인 tyrosinase가 tyrosine의 산화를 촉진하는 과정을 방지하여 피부 미백효과를 나타낸다. 미백의 주요 기전은 크게 6개로 나누어지며, 그 중 대표적으로 사용되는 기전은 tyrosinase가 tyrosine의 산화를 촉진하는 것을 방지하는 것으로 이 과정에서 ROS가 사용되며, 항산화제는 이러한 산화과정을 방지함으로서 피부 미백을 촉진시킨다.In addition, in relation to healthy skin, whitening is one of the fields in which importance increases along with antioxidants. This is because the Cosmetics Act was enacted independently from the Pharmaceutical Affairs Act in 1999, and since its implementation was finalized on July 1, 2000, research to find stable and effective whitening materials has been actively conducted. As a result, research on melanin production and control mechanisms has made progress, and based on these research results, various studies are being conducted to find materials with related activities in medicinal plants or natural products. Among them, there is a demand for the development of functional cosmetics using natural extracts that are relatively safer than synthetic substances that have side effects on the skin. In natural extracts, a representative component showing a whitening effect is polyphenol. Polyphenol is a substance found in all organisms and generally exhibits antioxidant activity. Antioxidant activity prevents tyrosinase, one of the main mechanisms of melanin production, from accelerating the oxidation of tyrosine, resulting in skin whitening effect. The main mechanisms of whitening are largely divided into 6, and the representative mechanism among them is to prevent tyrosinase from accelerating the oxidation of tyrosine. In this process, ROS are used, and antioxidants prevent this oxidation process, resulting in skin whitening. promotes
본 발명의 목적은 동물성 원료를 사용하지 않으면서도 우수한 항산화 및 미백 효능을 나타내어 기능성 비건 화장료의 원료로 사용될 수 있는 항산화 및 미백효능이 우수한 화장료용 조성물을 제공하는 것이다.An object of the present invention is to provide a cosmetic composition with excellent antioxidant and whitening effects that can be used as a raw material for functional vegan cosmetics by exhibiting excellent antioxidant and whitening effects without using animal raw materials.
본 발명의 목적은 와일드제라늄 추출물, 달맞이꽃 추출물, 감나무잎 추출물, 서양민들레잎 추출물, 한련초 추출물, 흰버드나무껍질 추출물 및 황금 추출물이 함유되는 것을 특징으로 하는 항산화 및 미백효능이 우수한 화장료용 조성물을 제공함에 의해 달성된다.An object of the present invention is to provide a cosmetic composition with excellent antioxidant and whitening effects, characterized in that it contains wild geranium extract, evening primrose extract, persimmon leaf extract, dandelion leaf extract, nasturtium extract, white willow bark extract and golden extract. is achieved by
본 발명의 바람직한 특징에 따르면, 상기 항산화 및 미백효능이 우수한 화장료용 조성물은 와일드제라늄 추출물 100 중량부, 달맞이꽃 추출물 80 내지 120 중량부, 감나무잎 추출물 80 내지 120 중량부, 서양민들레잎 추출물 80 내지 120 중량부, 한련초 추출물 80 내지 120 중량부, 흰버드나무껍질 추출물 80 내지 120 중량부 및 황금 추출물 80 내지 120 중량부로 이루어지는 것으로 한다.According to a preferred feature of the present invention, the cosmetic composition having excellent antioxidant and whitening effects includes 100 parts by weight of wild geranium extract, 80 to 120 parts by weight of evening primrose extract, 80 to 120 parts by weight of persimmon leaf extract, and 80 to 120 parts by weight of dandelion leaf extract. parts by weight, 80 to 120 parts by weight of nasturtium extract, 80 to 120 parts by weight of white willow bark extract and 80 to 120 parts by weight of golden extract.
본 발명의 더 바람직한 특징에 따르면, 상기 와일드제라늄 추출물, 달맞이꽃 추출물, 감나무잎 추출물, 서양민들레잎 추출물, 한련초 추출물, 흰버드나무껍질 추출물 및 황금 추출물은 열수 추출을 통해 제조되며.According to a more preferred feature of the present invention, the wild geranium extract, evening primrose extract, persimmon leaf extract, dandelion leaf extract, nasturtium extract, white willow bark extract and golden extract are prepared through hot water extraction.
상기 열수 추출은 정제수 100 중량부에 와일드제라늄, 달맞이꽃, 감나무잎, 서양민들레잎, 한련초, 흰버드나무껍질 및 황금으로 이루어진 그룹에서 선택된 하나로 이루어진 원물을 0.5 내지 1.5 중량부 혼합하고 75 내지 85℃의 온도에서 30 내지 90분 동안 이루어지는 것으로 한다.The hot water extraction is performed by mixing 0.5 to 1.5 parts by weight of a raw material selected from the group consisting of wild geranium, evening primrose, persimmon leaf, western dandelion leaf, nasturtium, white willow bark and gold in 100 parts by weight of purified water and mixing 75 to 85 ° C. It is made for 30 to 90 minutes at the temperature.
또한, 본 발명의 목적은 상기 항산화 및 미백효능이 우수한 화장료용 조성물이 31.25 내지 990㎍/mL로 함유하는 것을 특징으로 하는 항산화 및 미백효능이 우수한 화장료를 제공함에 의해서도 달성될 수 있다.In addition, the object of the present invention can also be achieved by providing a cosmetic composition with excellent antioxidant and whitening effects, characterized in that the cosmetic composition with excellent antioxidant and whitening effects contains 31.25 to 990 μg / mL.
본 발명의 바람직한 특징에 따르면, 상기 항산화 및 미백효능이 우수한 화장료는 비누, 클렌징 폼, 클렌징 크림, 클렌징 워터, 목욕제, 스킨, 로숀, 크림, 에센스, 토너 및 바디클렌져로 이루어진 그룹에서 선택된 하나의 제형으로 제조되는 것으로 한다.According to a preferred feature of the present invention, the cosmetic with excellent antioxidant and whitening effects is 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 It is assumed that it is manufactured with
본 발명에 따른 항산화 및 미백효능이 우수한 화장료용 조성물은 동물성 원료를 사용하지 않으면서도 우수한 항산화 및 미백 효능을 나타내어 기능성 비건 화장료의 원료로 사용될 수 있는 탁월한 효과를 나타낸다.The cosmetic composition having excellent antioxidant and whitening effects according to the present invention exhibits excellent antioxidant and whitening effects without using animal raw materials, thereby exhibiting excellent effects that can be used as raw materials for functional vegan cosmetics.
도 1은 본 발명의 실시예 1을 통해 제조된 항산화 및 미백효능이 우수한 화장료용 조성물의 DPPH 라디컬 소거능을 측정하여 나타낸 그래프이다.
도 2는 본 발명의 실시예 1을 통해 제조된 항산화 및 미백효능이 우수한 화장료용 조성물의 ABTS 라디컬 소거능을 측정하여 나타낸 그래프이다.1 is a graph showing DPPH radical scavenging ability of a cosmetic composition having excellent antioxidant and whitening effects prepared in Example 1 of the present invention.
2 is a graph showing the ABTS radical scavenging ability of the cosmetic composition having excellent antioxidant and whitening effects prepared in 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.
또한, 본 발명에서 "추출물"이란 추출 방법, 추출 용매, 추출된 성분 및 형태를 불문하고 천연물의 유효성분을 뽑아내어 얻어진 물질을 의미한다.In addition, in the present invention, "extract" refers to a substance obtained by extracting an active ingredient of a natural product regardless of extraction method, extraction solvent, extracted component and form.
추출방법으로는 열수 수출, 초음파 추출법 및 여과법, 환류 추출법 등이 있으며, 여러 추출방법이 병용되어 수행 될 수도 있으나 단독으로 수행될 수도 있다.Extraction methods include hot water export, ultrasonic extraction and filtration, and reflux extraction, and several extraction methods may be used in combination or may be performed alone.
바람직하게는 상기 7가지 혼합추출물은 열수 추출 하였으나 이에 제한 되는 것은 아니다.Preferably, the above seven mixed extracts were hot water extracted, but are not limited thereto.
본 발명에 따른 항산화 및 미백효능이 우수한 화장료용 조성물은 와일드제라늄 추출물(Geranium maculatum Extract), 달맞이꽃 추출물(Oenothera biennis Flower Extract), 감나무잎 추출물(Diospyros kaki Leaf Extract), 서양민들레잎 추출물{Dandelion (Taraxacum officinale) Leaf Extract}, 한련초 추출물(Eclipta prostrata Extract), 흰버드나무껍질 추출물(Salix alba Bark Extract) 및 황금 추출물(Scutellaria baicalensis Root Extract)이 함유되며, 와일드제라늄 추출물 100 중량부, 달맞이꽃 추출물 80 내지 120 중량부, 감나무잎 추출물 80 내지 120 중량부, 서양민들레잎 추출물 80 내지 120 중량부, 한련초 추출물 80 내지 120 중량부, 흰버드나무껍질 추출물 80 내지 120 중량부 및 황금 추출물 80 내지 120 중량부로 이루어지는 것이 바람직하다.The cosmetic composition having excellent antioxidant and whitening effects according to the present invention includes wild geranium extract (Geranium maculatum Extract), evening primrose extract (Oenothera biennis Flower Extract), persimmon leaf extract (Diospyros kaki Leaf Extract), western dandelion leaf extract {Dandelion (Taraxacum officinale) Leaf Extract}, nasturtium extract (Eclipta prostrata Extract), white willow bark extract (Salix alba Bark Extract) and golden extract (Scutellaria baicalensis Root Extract),
상기와 같이 7가지의 성분으로 이루어진 혼합 추출물은 개별 추출물 내지 2 내지 3가지의 조합추출물 대비 피부 미백 효과가 월등하게 향상된다.As described above, the mixed extract composed of 7 components significantly improves the skin whitening effect compared to individual extracts or a combination extract of 2 to 3 components.
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.
상기 달맞이꽃 추출물(Oenothera biennis Flower Extract)은 80 내지 120 중량부가 함유되며, 우수한 항산화 활성 및 세포사멸과 DNA 결실을 보호하는 효과를 나타낼 수 있을 뿐만 아니라, 오염물질에 의한 세포 사멸, 세포 증식 저해 및 피부상태를 개선하는 효과를 나타낸다.The evening primrose extract (Oenothera biennis Flower Extract) contains 80 to 120 parts by weight, and can exhibit excellent antioxidant activity and an effect of protecting cell death and DNA deletion, as well as inhibiting cell death by pollutants, cell proliferation and skin It shows the effect of improving the condition.
상기 감나무잎 추출물(Diospyros kaki Leaf Extract)은 80 내지 120 중량부가 함유되며, 감잎에는 다량의 Vitamin C가 함유되어 항산화 효과와 더불어 피부 미백 효과를 나타내며, 피부 탄력을 유지시키는 콜라겐의 합성을 유도해 노화를 막고 기미나 주근깨의 원인이되는 멜라닌 색소를 제거해 준다. 또한, 감나무잎에 함유된 Tannins성분들은 피부에 지혈성 수렴 작용을 하므로 피부에 탄력과 함께 매끈한 피부를 제공하며, 다양한 flavonoid 배당체와 phenol류 coumarin류 화합물이 다량 함유되어 항염, 항균 효과를 나타내면서 상처를 아물게 함과 동시에 균을 제거함으로써 깨끗하고 건강한 피부를 제공하는 역할을 한다.The persimmon leaf extract (Diospyros kaki Leaf Extract) contains 80 to 120 parts by weight, and persimmon leaves contain a large amount of Vitamin C, which shows an antioxidant effect and a skin whitening effect, and induces the synthesis of collagen that maintains skin elasticity to prevent aging. and removes the melanin pigment that causes spots and freckles. In addition, tannins contained in persimmon leaves have a hemostatic and astringent action on the skin, so they provide smooth skin with elasticity to the skin, and various flavonoid glycosides and phenols and coumarin compounds are contained in large amounts to show anti-inflammatory and antibacterial effects while reducing wounds. It heals and removes germs at the same time to provide clean and healthy skin.
상기 서양민들레잎 추출물{(Dandelion (Taraxacum officinale) Leaf Extract}은 80 내지 120 중량부가 함유되며, 항산화, 보습, 콜라겐 합성 촉진 효과가 우수하고, 그에 따라 피부 노화 예방 및 피부주름 개선, 피부 유연성 증진 및 탄력개선에 도움을 주며 피부 상처의 자연적 치유에도 도움을 주어 깨끗한 피부를 제공하는 역할을 한다.The dandelion leaf extract ( Dandelion (Taraxacum officinale) Leaf Extract} contains 80 to 120 parts by weight, and has excellent antioxidant, moisturizing, and collagen synthesis promoting effects, thereby preventing skin aging, improving skin wrinkles, and enhancing skin flexibility. And it helps to improve elasticity and helps to heal skin wounds naturally, so it plays a role in providing clean skin.
상기 한련초 추출물(Eclipta prostrata Extract)은 80 내지 120 중량부가 함유되며, 피부염을 개선하고, 모발 성장을 촉진하며, 피부가 갈라지는 피부염 등을 개선하여 깨끗하고 맑은 피부를 제공하는 역할을 한다.The nasturtium extract (Eclipta prostrata Extract) contains 80 to 120 parts by weight, and serves to improve dermatitis, promote hair growth, and improve dermatitis in which the skin is cracked to provide clean and clear skin.
상기 흰버드나무껍질 추출물{Salix Alba(Willow) Bark Extract}은 80 내지 120 중량부가 함유되며, 흰버드나무껍질에는 탄닌과 살리실산(바하,BHA)가 함유되어 있어 피부의 각질을 부드럽게 연화시켜 문제성 피부를 진정시키며, 피부속 수분 증발을 막아 피부의 보습력을 유지하는 역할을 한다.The white willow bark extract {Salix Alba (Willow) Bark Extract} contains 80 to 120 parts by weight, and the white willow bark contains tannin and salicylic acid (BHA, BHA), which gently softens the dead skin cells of the skin to treat problem skin. It soothes the skin and prevents the evaporation of moisture in the skin to maintain the moisturizing power of the skin.
상기 황금 추출물(Scutellaria baicalensis Root Extract)은 80 내지 120 중량부가 함유되며, 피부보습 및 항균효과를 부여하여 맑고 촉촉한 피부를 제공하는 역할을 한다.The golden extract (Scutellaria baicalensis Root Extract) contains 80 to 120 parts by weight, and serves to provide clear and moist skin by providing skin moisturizing and antibacterial effects.
상기의 와일드제라늄 추출물, 달맞이꽃 추출물, 감나무잎 추출물, 서양민들레잎 추출물, 한련초 추출물, 흰버드나무껍질 추출물 및 황금 추출물은 용매 추출이나 열수 추출을 통해 제조될 수 있는데, 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.The wild geranium extract, evening primrose extract, persimmon leaf extract, western dandelion leaf extract, nasturtium extract, white willow bark extract and golden extract may be prepared through solvent extraction or hot water extraction, wherein the solvent is C 1 to C 4 of lower alcohol, butylene glycol, propylene glycol, glycerin, hydrous ethanol, hydrous butylene glycol, hydrous propylene glycol, hexane, and ethyl acetate, and may consist of one or more selected from the group consisting of C 1 to C 4 lower alcohol It is preferred to use methanol or ethanol.
또한, 상기 열수 추출은 정제수 100 중량부에 와일드제라늄, 달맞이꽃, 감나무잎, 서양민들레잎, 한련초, 흰버드나무껍질 및 황금으로 이루어진 그룹에서 선택된 하나로 이루어진 원물 0.5 내지 1.5 중량부를 혼합하고 70 내지 130℃, 바람직하게는 80 내지 120℃, 더욱 바람직하게는 90 내지 110℃일 수 있으나 이에 제한되는 것은 아니다.In addition, the hot water extraction is performed by mixing 0.5 to 1.5 parts by weight of a raw material selected from the group consisting of wild geranium, evening primrose, persimmon leaf, dandelion leaf, nasturtium, white willow bark and gold in 100 parts by weight of purified water and mixing at 70 to 130 ° C. , Preferably from 80 to 120 ℃, more preferably from 90 to 110 ℃ may be, but is not limited thereto.
또한, 추출방법은 가열추출기에서 1 내지 5시간, 바람직하게는 2 내지 4시간, 더욱 바람직하게는 2시간 30분 내지 3시간 30분 동안 열수 추출하고, 이를 동결건조하여 수득하는 것일 수도 있으나 이에 제한되는 것은 아니다.In addition, the extraction method may be obtained by hot water extraction in a heat 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 going to be
상기의 성분으로 이루어지는 항산화 및 미백효능이 우수한 화장료용 조성물은 31.25 내지 990㎍/mL의 함량으로 화장품 베이스에 함유되어 항산화 및 미백효능이 우수한 화장료를 제공할 수 있는데, 상기 화장료에 함유된 항산화 및 미백효능이 우수한 화장료용 조성물의 함량이 31.25㎍/mL 미만이면, 화장료의 항산화 및 미백효능이 저하되며, 상기 항산화 및 미백효능이 우수한 화장료용 조성물의 함량이 990㎍/mL를 초과하게 되면 화장료의 항산화 및 미백효능은 크게 향상되지 않으면서 피부 독성이 증가하여 피부트러블을 유발할 수 있고, 제조비용을 지나치게 증가시키기 때문에 바람직하지 못하다.The cosmetic composition having excellent antioxidant and whitening effects comprising the above components can be contained in a cosmetic base in an amount of 31.25 to 990 μg/mL to provide a cosmetic having excellent antioxidant and whitening effects. If the content of the cosmetic composition with excellent efficacy is less than 31.25 μg/mL, the antioxidant and whitening efficacy of the cosmetic is reduced, and if the content of the cosmetic composition with excellent antioxidant and whitening efficacy exceeds 990 μg/mL, the antioxidant of the cosmetic is reduced. And whitening efficacy is not greatly improved, but skin toxicity is increased, which can cause skin trouble, and manufacturing cost is undesirably increased.
또한, 본 발명의 화장료 조성물은 통상적으로 화장료 조성물에서 추가할 수 있는 보조제를 하나 이상 더 포함 할 수 있다. 예를 들어 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.In addition, the cosmetic composition of the present invention may further include one or more adjuvants that can 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 발명인 화장료 조성물은 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.The cosmetic composition of the present invention can be prepared in any formulation commonly prepared in the field, for example, skin, lotion, essence, cream, cleansing, pack, powder, spray, etc., but is not limited thereto not.
예를 들어 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 클렌징 오일, 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, 립패치) 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) used for point makeup. It can be.
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.
이하에서는, 본 발명에 따른 항산화 및 미백효능이 우수한 화장료용 조성물의 제조방법 및 그 제조방법으로 제조된 화장료용 조성물의 물성을 실시예를 들어 설명하기로 한다.Hereinafter, the manufacturing method of the cosmetic composition having excellent antioxidant and whitening effects according to the present invention and the physical properties of the cosmetic composition prepared by the manufacturing method will be described with examples.
하지만, 이들은 본 발명을 보다 상세하게 설명하기 위한 것일 뿐 본 발명의 권리범위가 이에 한정되는 것은 아니다.However, these are only for explaining the present invention in more detail and the scope of the present invention is not limited thereto.
<제조예 1> 와일드제라늄 추출물의 제조<Preparation Example 1> Preparation of wild geranium extract
정제수 100 중량부에 와일드 제라늄 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.
<제조예 2> 달맞이꽃 추출물의 제조<Preparation Example 2> Preparation of evening primrose extract
정제수 100 중량부에 달맞이꽃 1 중량부를 혼합하고 80℃의 온도에서 60분 동안 가열한 후에 고형분을 걸러내어 달맞이꽃 추출물을 제조하였다.Evening primrose extract was prepared by mixing 1 part by weight of evening primrose with 100 parts by weight of purified water, heating at a temperature of 80 ° C. for 60 minutes, and filtering out the solid content.
<제조예 3> 감나무잎 추출물의 제조<Preparation Example 3> Preparation of persimmon leaf extract
정제수 100 중량부에 감나무잎 1 중량부를 혼합하고 80℃의 온도에서 60분 동안 가열한 후에 고형분을 걸러내어 감나무잎 추출물을 제조하였다.Persimmon leaf extract was prepared by mixing 1 part by weight of persimmon leaf with 100 parts by weight of purified water, heating at 80° C. for 60 minutes, and then filtering out the solid content.
<제조예 4> 서양민들레잎 추출물의 제조<Preparation Example 4> Preparation of dandelion leaf extract
정제수 100 중량부에 서양민들레잎 1 중량부를 혼합하고 80℃의 온도에서 60분 동안 가열한 후에 고형분을 걸러내어 서양민들레잎 추출물을 제조하였다.After mixing 1 part by weight of dandelion leaf with 100 parts by weight of purified water and heating at a temperature of 80 ° C. for 60 minutes, the solid content was filtered to prepare an extract of dandelion leaf.
<제조예 5> 한련초 추출물의 제조<Preparation Example 5> Preparation of nasturtium extract
정제수 100 중량부에 한련초 1 중량부를 혼합하고 80℃의 온도에서 60분 동안 가열한 후에 고형분을 걸러내어 한련초 추출물을 제조하였다.After mixing 1 part by weight of nasturtium with 100 parts by weight of purified water and heating at a temperature of 80 ° C. for 60 minutes, the solid content was filtered to prepare an extract of nasturtium.
<제조예 6> 흰버드나무껍질 추출물의 제조<Preparation Example 6> Preparation of white willow bark extract
정제수 100 중량부에 흰버드나무껍질 1 중량부를 혼합하고 80℃의 온도에서 60분 동안 가열한 후에 고형분을 걸러내어 흰버드나무껍질 추출물을 제조하였다.1 part by weight of white willow bark was mixed with 100 parts by weight of purified water, heated at 80° C. for 60 minutes, and then the solid content was filtered to prepare white willow bark extract.
<제조예 7> 황금 추출물의 제조<Preparation Example 7> Preparation of golden extract
정제수 100 중량부에 황금 1 중량부를 혼합하고 80℃의 온도에서 60분 동안 가열한 후에 고형분을 걸러내어 황금 추출물을 제조하였다.A golden extract was prepared by mixing 1 part by weight of gold with 100 parts by weight of purified water, heating at 80 ° C. for 60 minutes, and then filtering out the solid content.
<실시예 1><Example 1>
상기 제조예 1 내지 7을 통해 제조된 와일드제라늄 추출물, 달맞이꽃 추출물, 감나무잎 추출물, 서양민들레잎 추출물, 한련초 추출물, 흰버드나무껍질 추출물 및 황금 추출물을 동일한 중량으로 혼합하여 항산화 및 미백효능이 우수한 화장료용 조성물을 제조하였다.Wild geranium extract, evening primrose extract, persimmon leaf extract, western dandelion leaf extract, nasturtium extract, white willow bark extract and golden extract prepared in Preparation Examples 1 to 7 were mixed in the same weight to make cosmetics with excellent antioxidant and whitening effects A composition was prepared.
상기 실시예 1을 통해 제조된 항산화 및 미백효능이 우수한 화장료용 조성물의 물성을 측정하여 아래 표 1 내지 3 및 도 1 내지 2에 나타내었다.The physical properties of the cosmetic composition having excellent antioxidant and whitening effects prepared in Example 1 were measured and are shown in Tables 1 to 3 and FIGS. 1 to 2 below.
{이때, 항산화능 실험 및 세포실험을 위해 동결건조를 통해 추출물의 건조중량을 측정하였으며, 얻어진 추출물은 증류수로 1 mg/mL 농도로 재용해시킨 뒤 동일 비율로 혼합하였다. 이후 syringe filter(PVDF, 0.2 μm를 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 the cell test, and the obtained extract was re-dissolved in distilled water at a concentration of 1 mg/mL and then mixed in the same ratio. Afterwards, bacteria and insoluble matter were removed through a syringe filter (PVDF, 0.2 μm).
또한, 상기 화장료용 조성물의 물성은 DPPH 라디컬 소거능, ABTS 라디컬 소거능, FRAP, 폴리페놀 함량, 플라보노이드 함량, 세포 독성 및 멜라닌 생성을 측정하는 방법을 이용하였다.In addition, the physical properties of the cosmetic composition used a method for measuring DPPH radical scavenging ability, ABTS radical scavenging ability, FRAP, polyphenol content, flavonoid content, cytotoxicity and melanin production.
- DPPH 라디컬 소거능 측정- Measurement of DPPH radical scavenging ability
DPPH assay는 DPPH의 라디컬 활성이 항산화제에 의해 소거되면 원래의 520 nm 파장의 빛에 대한 흡광도가 감소하는 것을 이용하는 방법으로, DPPH 용액을 만들기 위해 70% ethanol을 용매로, DPPH를 1%(w/v)가 되도록 녹인 뒤 filter paper를 이용해 불용된 DPPH를 제거하였다. 이후 용액의 520 nm 흡광도가 1.00이 되도록 70% ethanol로 다시 희석하여 DPPH 용액으로 사용하였다. 500 μg/mL를 기준으로 2 배씩 희석하여 만들어진 화장료용 조성물과 1.0 mL와 DPPH 용액 1.0 mL를 혼합 후 30 분간 반응시켰다. 그 후 520 nm에서의 흡광도를 측정하였다. 이때, 화장료용 조성물 및 환원된 DPPH 용액의 흡광도를 측정하여 실험결과에서 제외했다. DPPH 실험결과는 ascorbic acid의 IC50를 측정 후 비교하여 항산화능을 평가하였다.The DPPH assay uses the decrease in absorbance of the original 520 nm wavelength light when the radical activity of DPPH is quenched by antioxidants. w/v), and undissolved DPPH was removed using filter paper. Then, the solution was diluted with 70% ethanol so that the absorbance at 520 nm was 1.00, and used as a DPPH solution. After mixing 1.0 mL of the cosmetic composition prepared by diluting 500 μg/mL by 2 times and 1.0 mL of DPPH solution, the mixture was reacted for 30 minutes. After that, the absorbance at 520 nm was measured. At this time, the absorbance of the cosmetic composition and the reduced DPPH solution was measured and excluded from the experimental results. The DPPH test results evaluated the antioxidant ability by measuring and comparing the IC 50 of ascorbic acid.
- ABTS 라디컬 소거능 측정- Measurement of ABTS radical scavenging ability
ABTS assay는 ABTS의 라디컬 활성이 항산화제에 의해 소거되면 원래의 740 nm 파장의 빛에 대한 흡광도가 감소하는 것을 이용하는 방법으로, ABTS 용액은 발색 되지 않은 7 mM ABTS 용액에 2.45 mM potassium persulfate를 녹여 12 시간 동안 발색반응을 진행시켰다. 이후 발색이 끝난 ABTS 용액의 740 nm에서의 흡광도를 측정하여 흡광도가 0.7이 되도록 희석하여 사용하였다. 500 μg/mL를 기준으로 2 배씩 희석하여 만들어진 다양한 농도의 화장료용 조성물 1.0 mL와 ABTS 용액 1.0 mL를 혼합 후 30 분간 반응시킨 후 740 nm에서의 흡광도를 측정하였다. 이때 추출물 및 환원된 ABTS 용액의 흡광도를 측정하여 실험결과에서 제외했다. ABTS 실험결과는 ascorbic acid의 IC50를 측정 후 비교하여 항산화능을 평가하였다.The ABTS assay uses the fact that the absorbance of the original 740 nm wavelength light decreases when the radical activity of ABTS is quenched by an antioxidant. The color development reaction proceeded for 12 hours. Then, the absorbance of the ABTS solution after color development was measured at 740 nm, and the absorbance was diluted to 0.7. After mixing 1.0 mL of the cosmetic composition of various concentrations prepared by diluting 500 μg / mL by 2 times and 1.0 mL of ABTS solution, the mixture was reacted for 30 minutes, and then 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. Antioxidant activity was evaluated by comparing the results of the ABTS test after measuring the IC 50 of ascorbic acid.
- FRAP 측정- FRAP measurement
실험에는 화장료용 조성물을 0.5 mg/mL 농도로 희석하여 사용하였다. 화장료용 조성물 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 20 분 동안 반응시켰다. 이 후 반응이 끝난 시료는 10% trichloroacetic acid 2.5 ml를 넣은 뒤 원심분리(3000 × G, 15 min)하여 침전물을 제거하였으며, 침전물이 제거된 용액 1 mL와 0.1% ferric chloride 용액 0.2 mL를 혼합시켰다. 그 후 700 nm에서 흡광도를 측정하여 환원력을 평가하였다.In the experiment, the cosmetic composition was diluted to a concentration of 0.5 mg/mL and used. After mixing 2.5 mL of the cosmetic composition 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 was completed, 2.5 ml of 10% trichloroacetic acid was added to the sample, and the precipitate was removed by centrifugation (3000 × G, 15 min), and 1 mL of the solution from which the precipitate was removed was mixed with 0.2 mL of 0.1% ferric chloride solution. . After that, the absorbance was measured at 700 nm to evaluate the reducing power.
이때, FRAP은 ascorbic acid를 기준으로 standard curve를 작성하여 측정하였다.At this time, FRAP was measured by preparing a standard curve based on ascorbic acid.
- 폴리페놀 함량 분석- Analysis of polyphenol content
폴리페놀 함량 분석은 Folin-Cioc
- 플라보노이드 함량 분석- Analysis of flavonoid content
플라보노이드 함량 분석은 플라보노이드와 알루미늄 이온의 결합에 따른 발색 반응 차이를 통해 측정하였다. 상기 화장료용 조성물 1.0 mL에 NaNO2 (5%, w/v) 0.3 mL를 주입한 뒤 5 분 동안 실온에서 방치한 후 AlCl3 (2%, w/v) 0.5 mL를 주입한 후 반응이 일어날 수 있도록 6 분간 방치하였다. 마지막으로 1 M NaOH 0.5 mL를 주입하여 중화시킨 뒤 510 nm에서의 흡광도를 측정하였다. 플라보노이드 농도는 quercetin을 기준으로 standard curve를 이용해 환산하였다.Analysis of flavonoid content was measured through the difference in color reaction according to the combination of flavonoid and aluminum ion. After injecting 0.3 mL of NaNO 2 (5%, w/v) into 1.0 mL of the cosmetic composition, injecting 0.5 mL of AlCl 3 (2%, w/v) after leaving it at room temperature for 5 minutes, the reaction occurs. It was allowed to stand for 6 minutes. Finally, 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 based on quercetin.
- 세포 배양- Cell culture
세포 독성 실험에는 B16F10 melanoma cell을 사용하였다. 세포 배양 배지로는 DMEM (Dulbecco's Modified Eagle Medium, GE healthcare, USA)를 사용하였으며, FBS (fetal bovine serum, Sigma, USA) 5%와 항생물질로서 Penicillin-Streptomycin (100X) (Sigma, USA)를 첨가하여 제조하였다. Melanin 생성 유도 물질로는 α-melanocyte stimulating hormone (α-MSH, Sigma, USA)을 사용하였다. 추출물은 DMSO에 용해시켜 처리하였다.B16F10 melanoma cells were used for cytotoxicity experiments. DMEM (Dulbecco's Modified Eagle Medium, GE healthcare, USA) was used as the cell culture medium, and 5% FBS (fetal bovine serum, Sigma, USA) and Penicillin-Streptomycin (100X) (Sigma, USA) were added as an antibiotic. It was manufactured. As a melanin production inducing substance, α-melanocyte stimulating hormone (α-MSH, Sigma, USA) was used. The extract was treated by dissolving in DMSO.
- 세포 독성 측정- Measurement of cytotoxicity
세포 독성의 측정 방법으로는 MTT assay를 실시하였다. 전배양된 B16F10을 96 well plate에 well 당 5×104 cell을 주입하여 24 시간 배양하였다. 배양이 끝난 후 100 μg/mL를 기준으로 2 배 다단희석을 통해 6.25-100 μg/mL 농도로 희석한 7가지 추출물을 처리하여 48 시간 동안 배양하였다. 2차 배양 후 상층 배양액을 제거하고 MTT 용액(5 mg/mL)를 가해준 뒤 온도 37℃, CO2 농도 5% 의 환경에서 MTT를 결정이 생성되도록 하였다. 각 well에 생성된 결정은 DMSO로 녹여 540 nm 에서의 흡광도를 측정하였다.As a method for measuring cytotoxicity, MTT assay was performed. The pre-cultured B16F10 was injected into a 96 well plate at 5×10 4 cells per well and cultured for 24 hours. After the incubation, 7 extracts diluted to a concentration of 6.25-100 μg/mL through 2-fold multi-dilution based on 100 μg/mL were treated and cultured for 48 hours. After the secondary culture, the supernatant culture medium was removed, MTT solution (5 mg/mL) was added, and MTT crystals were formed in an environment of 37° C. and 5% CO2 concentration. Crystals generated in each well were dissolved in DMSO and absorbance at 540 nm was measured.
- 멜라닌 생성 측정- Measurement of melanin production
B16F10 melanoma cell을 사용하여 멜라닌 생성량 측정 실험을 진행하였다. 세포 배양 배지로는 DMEM broth를 사용하였으며, FBS 5%와 항생물질로서 Penicillin-Streptomycin (100X)를 첨가하여 제조하였다. 화장료용 조성물은 DMSO에 용해시켜 처리하였다. Melanin 생성 유도 물질로는 α-melanocyte stimulating hormone (α-MSH, Sigma, USA)을 사용하였다. 혼합물은 DMSO에 용해시켜 처리하였다.An experiment was conducted to measure the amount of melanin produced using B16F10 melanoma cells. DMEM broth was used as a cell culture medium, and it was prepared by adding 5% FBS and Penicillin-Streptomycin (100X) as an antibiotic. The cosmetic composition was treated by dissolving in DMSO. As a melanin production inducing substance, α-melanocyte stimulating hormone (α-MSH, Sigma, USA) was used. The mixture was treated by dissolving in DMSO.
먼저 배양된 B16F10을 96 well plate에 well 당 5×104 cell을 주입하여 24 시간 배양하였다. 배양이 끝난 후 100 μg/mL를 기준으로 2 배 다단희석을 통해 다양한 농도로 제조된 상기 화장료용 조성물과 α-MSH 100 nM을 처리하여 72 시간 동안 배양하였다. 배양 후 1N NaOH 0.1 μL 처리 후 1시간 동안 방치하여 멜라닌을 용해시킨 후 405nm 흡광도를 측정하였다.}First, the cultured B16F10 was cultured for 24 hours by injecting 5×10 4 cells per well into a 96 well plate. After the incubation was completed, the cosmetic composition prepared at various concentrations through 2-fold multi-dilution based on 100 μg/mL was treated with 100 nM of α-MSH and cultured for 72 hours. After incubation, 1N NaOH was treated with 0.1 μL, left for 1 hour to dissolve melanin, and then 405 nm absorbance was measured.}
상기 실시예 1을 통해 제조된 화장료용 조성물의 DPPH 라디컬 소거능을 측정하여 아래 도 1에 나타내었다.The DPPH radical scavenging ability of the cosmetic composition prepared in Example 1 was measured and shown in FIG. 1 below.
측정 전 비교를 위해 ascorbic acid의 EC50 의 항산화 수치를 측정하였으며, 각 농도를 2.5-40 μg/mL 범위로 반응시켜 얻어진 ascorbic acid의 EC50 수치는 17.44 μg/mL였다.For comparison before measurement, the antioxidant value of EC 50 of ascorbic acid was measured, and the EC 50 value of ascorbic acid obtained by reacting each concentration in the range of 2.5-40 μg/mL was 17.44 μg/mL.
화장료용 조성물을 31.25 내지 500 μg/mL로 희석하여 실험을 진행한 결과는 아래 도 1에 나타낸 바와 같이 농도 31.25 μg/mL 에서 16.88±3.42%, 62.5 μg/mL 에서 33.99±2.95%, 125 μg/mL 에서 57.13±0.62%, 250 μg/mL 에서 89.94±2.13%, 500 μg/mL 에서 96.43±1.40%의 DPPH 라디칼 소거능을 보였다. 실험결과 EC50을 통해 비교 확인한 결과 ascorbic acid 는 17.44 μg/mL, 화장료용 조성물은 105.74 μg/mL로 나타나 화장료용 조성물이 ascorbic acid과 비교 시 약 16.50%의 항산화능이 있음을 확인하였다.The results of the experiment by diluting the cosmetic composition to 31.25 to 500 μg/mL were 16.88±3.42% at a concentration of 31.25 μg/mL, 33.99±2.95% at 62.5 μg/mL, and 125 μg/mL, as shown in Figure 1 below. DPPH radical scavenging activity was 57.13±0.62% at mL, 89.94±2.13% at 250 μg/mL, and 96.43±1.40% at 500 μg/mL. As a result of comparison and confirmation through EC 50 as a result of the experiment, ascorbic acid was 17.44 μg/mL and that of the cosmetic composition was 105.74 μg/mL, confirming that the cosmetic composition had an antioxidant capacity of about 16.50% compared to that of ascorbic acid.
본 발명에서 사용된 원료 중 하나인 서양민들레 추출물의 DPPH 라디컬 소거능과 비교할 경우 서양민들레 열수 추출물의 EC50은 176.99 μg/mL로 화장료용 조성물의 항산화능이 약 1.67배 강한 것으로 나타났다. 한편 그라비올라 잎 추출물에 대한 연구의 결과를 기반으로 EC50을 계산한 결과 화장료용 조성물의 EC50은 875.98 μg/mL로 나타나 화장료용 조성물의 항산화능이 약 8.28배 강한 것을 알 수 있다(아래 도 1에서 * p<.05, ** p<.01, *** p<.001).Compared to the DPPH radical scavenging activity of the dandelion extract, which is one of the raw materials used in the present invention, the EC 50 of the dandelion hot water extract was 176.99 μg/mL, indicating that the antioxidant activity of the cosmetic composition was about 1.67 times stronger. On the other hand, as a result of calculating the EC 50 based on the results of the study on the Graviola leaf extract, the EC 50 of the cosmetic composition was 875.98 μg / mL, indicating that the antioxidant capacity of the cosmetic composition was about 8.28 times stronger (Fig. 1 below). in *p<.05, **p<.01, ***p<.001).
또한, 상기 실시예 1을 통해 제조된 화장료용 조성물의 ABTS 라디컬 소거능을 측정하여 아래 도 2에 나타내었다.In addition, the ABTS radical scavenging ability of the cosmetic composition prepared in Example 1 was measured and shown in FIG. 2 below.
아래 도 2에 나타낸 것처럼, g/mL 범위로 반응시켜 얻어진 ascorbic acid의 EC50 수치는 9.11 μg/mL였다. 화장료용 조성물을 31.25 내지 500 μg/mL로 희석하여 실험을 진행한 결과 31.25 μg/mL 에서 20.21±1.54%, 62.5 μg/mL 에서 41.22±1.58%, 125 μg/mL 에서 65.28±0.50%, 250 μg/mL 에서 92.78±2203%, 500 μg/mL 에서 99.21±1.14%의 ABTS 라디칼 소거능이 보였다. 실험결과를 IC50을 통해 비교 확인한 결과 ascorbic acid 는 9.11 μg/mL, 본 추출물은 85.31 μg/mL로 나타나 화장료용 조성물이 ascorbic acid과 비교 시 약 10.68%의 항산화능이 있음을 확인하였다.As shown in Figure 2 below, the EC 50 value of ascorbic acid obtained by reacting in the g / mL range was 9.11 μg / mL. As a result of diluting the cosmetic composition from 31.25 to 500 μg/mL and conducting experiments, 20.21±1.54% at 31.25 μg/mL, 41.22±1.58% at 62.5 μg/mL, 65.28±0.50% at 125 μg/mL, and 250 μg ABTS radical scavenging activity was 92.78±2203% at /mL and 99.21±1.14% at 500 μg/mL. As a result of comparing and confirming the experimental results through IC50, ascorbic acid was 9.11 μg/mL and this extract was 85.31 μg/mL, confirming that the cosmetic composition had about 10.68% antioxidant capacity compared to ascorbic acid.
Spiraea prunifolia 뿌리 열수추출물과 해당 결과를 비교할 경우 Spiraea prunifolia 뿌리 열수추출물의 EC50은 124.0 μg/mL로 화장료용 조성물의 항산화능이 약 1.45배 강한 것을 알 수 있다.(아래 도 2에서 * p<.05, ** p<.01, *** p<.001)When comparing the results with the hot water extract of Spiraea prunifolia root, the EC 50 of the hot water extract of Spiraea prunifolia root was 124.0 μg/mL, indicating that the antioxidant activity of the cosmetic composition was about 1.45 times stronger. (* p<.05 in FIG. 2 below) , **p<.01, ***p<.001)
또한, 상기 실시예 1을 통해 제조된 화장료용 조성물의 FRAP, 폴리페놀 농도(TPC) 및 플라보노이드 농도(TFC)를 측정하여 아래 표 1에 나타내었다.In addition, FRAP, polyphenol concentration (TPC) and flavonoid concentration (TFC) of the cosmetic composition prepared in Example 1 were measured and shown in Table 1 below.
<표 1><Table 1>
상기 표 1에 나타낸 것처럼, 상기 실시예 1을 통해 제조된 화장료용 조성물과 ascorbic acid의 철 이온에 대한 환원력인 FRAP을 측정하여 비교한 결과 화장료용 조성물의 1 mg의 FRAP은 ascorbic acid 198.01±5.50 μg의 FRAP과 같음을 알 수 있었다. 또한, 폴리페놀의 경우 30.19±0.75 μg/mg으로 나타났으며, 플라보노이드의 경우 9.12±0.36 μg/mg으로 나타났다.As shown in Table 1, as a result of measuring and comparing FRAP, which is the reducing power of iron ions of the cosmetic composition prepared in Example 1 and ascorbic acid, FRAP of 1 mg of the cosmetic composition is ascorbic acid 198.01 ± 5.50 μg was found to be identical to that of FRAP. In addition, in the case of polyphenols, it was 30.19 ± 0.75 μg / mg, and in the case of flavonoids, it was 9.12 ± 0.36 μg / mg.
상황버섯 추출물과 폴리페놀과 플라보노이드 농도를 비교할 경우 상황버섯 열수 추출물의 폴리페놀은 16.33 μg/mg으로 7가지 혼합추출물의 폴리페놀의 농도가 약 1.8배 높은 것으로 나타났다.Comparing the polyphenol and flavonoid concentrations of Phellinus Phellinus extract, the polyphenol content of the hot-water extract of Phellinus Phellinus was 16.33 μg/mg, which was about 1.8 times higher than the concentration of polyphenols in the 7 mixed extracts.
한편 플라보노이드 농도를 비교할 경우 상황버섯 열수 추출물의 플라보노이드는 4.50 μg/mg으로 화장료용 조성물의 플라보노이드의 농도가 약 2.0배 높은 것을 알 수 있다.On the other hand, when comparing the flavonoid concentrations, it can be seen that the flavonoids of the hot water extract of Phellinus Phellinus are 4.50 μg/mg, which is about 2.0 times higher than the concentration of flavonoids in the cosmetic composition.
또한, 상기 실시예 1을 통해 제조된 화장료용 조성물의 세포 독성을 측정하여 아래 표 2에 나타내었다. In addition, the cytotoxicity of the cosmetic composition prepared in Example 1 was measured and shown in Table 2 below.
{단, 화장료용 조성물이 세포에 미치는 독성을 평가하기 위해서 MTT assay를 이용한 세포 생존률을 측정하였는데, 화장료용 조성물은 배지에서의 최종농도가 6.25 내지 100 μg/mL가 되도록 배합하여 배양을 진행하였다.}{However, in order to evaluate the toxicity of the cosmetic composition to cells, the cell viability was measured using the MTT assay. The cosmetic composition was formulated and cultured so that the final concentration in the medium was 6.25 to 100 μg / mL. }
<표 2><Table 2>
상기 표 2에 나타낸 것처럼, 100 μg/mL 농도에서 90.06±2.83%, 50 μg/mL 농도에서 92.32±0.88%, 25 μg/mL 농도에서 94.00±1.08%, 12.5 μg/mL 농도에서 97.16±0.63%, 6.25 μg/mL 농도에서 99.23±1.64%의 세포 생존률이 나타났다(* p<.05, ** p<.01).As shown in Table 2 above, 90.06 ± 2.83% at 100 μg / mL concentration, 92.32 ± 0.88% at 50 μg / mL concentration, 94.00 ± 1.08% at 25 μg / mL concentration, 97.16 ± 0.63% at 12.5 μg / mL concentration , a cell viability of 99.23±1.64% was observed at the concentration of 6.25 μg/mL (* p<.05, ** p<.01).
대조군과 student‘s t test를 통해 비교한 결과 12.5 μg/mL부터 유의미한 세포독성을 보였다. 하지만 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 것으로 나타났다.As a result of comparison through the control group and student's t test, significant cytotoxicity was shown from 12.5 μg/mL. However, in accordance with ISO 10993-5 and Ministry of Food and Drug Safety Notice No. 2014-115, it is considered to be toxic when it exhibits cytotoxicity of 20% or more, and it was found that cytotoxicity did not appear at all experimental concentrations under these standards.
또한, 상기 실시예 1을 통해 제조된 화장료용 조성물의 미백 활성을 측정하여 아래 표 3에 나타내었다.In addition, the whitening activity of the cosmetic composition prepared in Example 1 was measured and shown in Table 3 below.
{단, 화장료용 조성물이 멜라닌 생성에 미치는 영향을 평가하기 위해서 B16F10의 멜라닌 생성량을 측정하였으며, 화장료용 조성물은 배지에서의 최종농도가 6.25 내지 100 μg/mL가 되도록 배합하여 배양을 진행하였다.}{However, in order to evaluate the effect of the cosmetic composition on melanin production, the amount of melanin production of B16F10 was measured, and the cosmetic composition was formulated and cultured so that the final concentration in the medium was 6.25 to 100 μg / mL.}
<표 3><Table 3>
상기 표 3에 나타낸 것처럼, 100 μg/mL 농도에서 34.70±2.97%, 50 μg/mL 농도에서 33.88±4.95%, 25 μg/mL 농도에서 32.73±6.89%, 12.5 μg/mL 농도에서 27.30±6.25%, 6.25 μg/mL 농도에서 23.36±4.19%의 멜라닌 생성 억제 효과가 나타났다(** p<.01).As shown in Table 3 above, 34.70 ± 2.97% at 100 μg / mL concentration, 33.88 ± 4.95% at 50 μg / mL concentration, 32.73 ± 6.89% at 25 μg / mL concentration, 27.30 ± 6.25% at 12.5 μg / mL concentration , 23.36±4.19% of melanin production inhibitory effect was shown at the concentration of 6.25 μg/mL (** p<.01).
상황버섯 추출물과 미백활성을 비교할 경우 열수 추출물에서 양쪽 최대 30% 이상의 미백효과를 내었으나 저농도 조건인 25 μg/mL에서의 결과를 비교했을 때, 상황버섯의 경우 21.46%의 미백효과를 보인 데 비해, 화장료용 조성물의 경우 32.73%의 미백효과를 보여 약 1.5배 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 , in the case of a cosmetic composition, it showed a whitening effect of 32.73%, which was about 1.5 times higher.
한편, 개별 추출물에 대한 예비 실험 결과 중 가장 높은 황금 열수 추출물의 경우 100 μg/mL 농도에서 28.31±1.57%의 미백효과를 보였으며, 이는 황금 열수 추출물의 항산화와 멜라닌생성 저해 효과에 대한 선행논문에서 농도에 따라 최대 29%의 멜라닌 생성 억제효과를 보인 것과 유사하다. 또한 본 발명에 따른 혼합 추출물은 황금 열수 추출물에 비해 약 6.39% 높은 효과를 보였으며, 혼합 추출물의 경우 황금 추출물의 비율이 낮음에도 황금추출물보다 높은 효과를 보여 상승효과가 나타난 것으로 판단된다.On the other hand, in the case of the gold hot water extract, which is the highest among preliminary experiments on individual extracts, at a concentration of 100 μg/mL, it showed a whitening effect of 28.31 ± 1.57%. It is similar to the one that showed up to 29% of melanin production inhibitory effect depending on the concentration. In addition, the mixed extract according to the present invention showed about 6.39% higher effect than the golden hot water extract, and in the case of the mixed extract, it is judged that the synergistic effect was shown by showing a higher effect than the golden extract even though the ratio of the golden extract was low.
따라서, 본 발명에 따른 항산화 및 미백효능이 우수한 화장료용 조성물은 동물성 원료를 사용하지 않으면서도 우수한 항산화 및 미백 효능을 나타내어 기능성 비건 화장료의 원료로 사용될 수 있다.Therefore, the cosmetic composition having excellent antioxidant and whitening effects according to the present invention can be used as a raw material for functional vegan cosmetics by showing excellent antioxidant and whitening effects without using animal raw materials.
Claims (5)
A cosmetic composition with excellent antioxidant and whitening effects, comprising wild geranium extract, evening primrose extract, persimmon leaf extract, dandelion leaf extract, nasturtium extract, white willow bark extract and golden extract.
상기 항산화 및 미백효능이 우수한 화장료용 조성물은 와일드제라늄 추출물 100 중량부, 달맞이꽃 추출물 80 내지 120 중량부, 감나무잎 추출물 80 내지 120 중량부, 서양민들레잎 추출물 80 내지 120 중량부, 한련초 추출물 80 내지 120 중량부, 흰버드나무껍질 추출물 80 내지 120 중량부 및 황금 추출물 80 내지 120 중량부로 이루어지는 것을 특징으로 하는 항산화 및 미백효능이 우수한 화장료용 조성물.
The method of claim 1,
The cosmetic composition having excellent antioxidant and whitening effects includes 100 parts by weight of wild geranium extract, 80 to 120 parts by weight of evening primrose extract, 80 to 120 parts by weight of persimmon leaf extract, 80 to 120 parts by weight of dandelion leaf extract, 80 to 120 parts by weight of nasturtium extract. A cosmetic composition with excellent antioxidant and whitening effects, characterized in that it consists of 80 to 120 parts by weight of white willow bark extract and 80 to 120 parts by weight of golden extract.
상기 와일드제라늄 추출물, 달맞이꽃 추출물, 감나무잎 추출물, 서양민들레잎 추출물, 한련초 추출물, 흰버드나무껍질 추출물 및 황금 추출물은 열수 추출을 통해 제조되며.
상기 열수 추출은 정제수 100 중량부에 와일드제라늄, 달맞이꽃, 감나무잎, 서양민들레잎, 한련초, 흰버드나무껍질 및 황금으로 이루어진 그룹에서 선택된 하나로 이루어진 원물 0.5 내지 1.5 중량부를 혼합하고 75 내지 85℃의 온도에서 30 내지 90분 동안 이루어지는 것을 특징으로 하는 항산화 및 미백효능이 우수한 화장료용 조성물.
The method of claim 1,
The wild geranium extract, evening primrose extract, persimmon leaf extract, western dandelion leaf extract, nasturtium extract, white willow bark extract and golden extract are prepared through hot water extraction.
The hot water extraction is performed by mixing 0.5 to 1.5 parts by weight of raw material selected from the group consisting of wild geranium, evening primrose, persimmon leaf, dandelion leaf, nasturtium, white willow bark and gold with 100 parts by weight of purified water and mixing at a temperature of 75 to 85 ° C. A cosmetic composition having excellent antioxidant and whitening effects, characterized in that it is made for 30 to 90 minutes in
A cosmetic composition having excellent antioxidant and whitening effects according to any one of claims 1 to 3, which is characterized in that it contains 31.25 to 990 μg/mL of the cosmetic composition having excellent antioxidant and whitening effects.
상기 항산화 및 미백효능이 우수한 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 및 미백효능이 우수한 화장료.The method of claim 4,
Antioxidant, characterized in that the cosmetics with excellent antioxidant and whitening effect 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 and cosmetics with excellent whitening effect.
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