KR102319686B1 - Composition for skin whitening or anti-wrinkle comprising fermented extract of Ambrosia trifida and Annual fleabane - Google Patents
Composition for skin whitening or anti-wrinkle comprising fermented extract of Ambrosia trifida and Annual fleabane Download PDFInfo
- Publication number
- KR102319686B1 KR102319686B1 KR1020190094289A KR20190094289A KR102319686B1 KR 102319686 B1 KR102319686 B1 KR 102319686B1 KR 1020190094289 A KR1020190094289 A KR 1020190094289A KR 20190094289 A KR20190094289 A KR 20190094289A KR 102319686 B1 KR102319686 B1 KR 102319686B1
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- South Korea
- Prior art keywords
- ragweed
- fermented
- extract
- maple leaf
- present
- Prior art date
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Abstract
본 발명은 단풍잎돼지풀 및 개망초 발효 추출물에 관한 것으로서, 더욱 상세하게는 상기 발효 추출물을 포함하는 피부 미백 또는 주름 개선용 조성물에 관한 것이다. 본 발명에 따른 단풍잎돼지풀 및 개망초 발효 추출물은 티로시나아제의 활성을 억제하고, 멜라닌 생성 세포에서 멜라닌 생성을 억제하는 효과를 가지고 있을 뿐만 아니라, 엘라스타아제의 저해 활성이 우수한바, 피부 미백 또는 주름 개선을 위한 화장료, 피부 외용제 및 식품 분야 등에서 다양하게 활용될 수 있다.The present invention relates to a fermented extract of Maple Leaf ragweed and Albaniasis, and more particularly, to a skin whitening or anti-wrinkle composition comprising the fermented extract. The fermented maple leaf ragweed extract according to the present invention inhibits the activity of tyrosinase and has the effect of inhibiting the production of melanin in melanin-producing cells, and has excellent inhibitory activity of elastase, skin whitening or It can be used in various fields such as cosmetics for wrinkle improvement, external preparations for skin, and food.
Description
본 발명은 단풍잎돼지풀 및 개망초 발효 추출물에 관한 것으로서, 더욱 상세하게는 상기 발효 추출물을 포함하는 피부 미백 또는 주름 개선용 조성물에 관한 것이다. The present invention relates to a fermented extract of Maple Leaf ragweed and Albaniasis, and more particularly, to a skin whitening or anti-wrinkle composition comprising the fermented extract.
산업화로 인한 환경오염은 일반 생활의 장애를 초래하는데, 특히 인체 보호막으로서 피부질환이 증가할 뿐 아니라 삶의 질 향상이라는 인식 변화로 인해 피부 관련 미용과 질환 완화 치료 소재에 대한 관심이 증가하고 있다. 일반적으로 환경공해의 심화와 함께 자외선에 의한 피부손상 등으로 하얀 피부에 대한 선호도가 높아지고 있고, 투명 화장으로 보다 맑고 깨끗한 피부를 추구함에 따라 피부 미백 효과를 얻을 수 있는 방법에 대한 관심이 고조되고 있다. 또한, 현대 가치관의 다양화와 개성화가 두드러지면서 상기와 같은 관심의 증가는 미백, 보습, 세포 재생, 주름개선 등의 성분을 함유한 기능성 화장품의 개발로 이어지고 있으며, 제품의 안전성을 지향하는 화장품 업계는 천연물 소재에 대한 연구개발을 활발하게 진행하고 있다.Environmental pollution caused by industrialization causes obstacles in general life. In particular, interest in skin-related beauty and disease alleviation treatment materials is increasing due to changes in the perception of improving quality of life as well as increasing skin diseases as a protective film for the human body. In general, the preference for white skin is increasing due to skin damage caused by ultraviolet rays along with the deepening of environmental pollution, and as people pursue clearer and cleaner skin with transparent makeup, interest in methods to obtain skin whitening effect is increasing. . In addition, as the diversification and individualization of modern values stand out, the increase in interest as above has led to the development of functional cosmetics containing ingredients such as whitening, moisturizing, cell regeneration, and wrinkle improvement, and the cosmetic industry aiming for product safety is actively conducting R&D on natural materials.
신체의 가장 바깥 부분에 위치하는 표피는 기저층, 유극층, 각질층의 순서로 배열되어 있다. 이를 구성하는 다양한 세포 종류 중 멜라닌 세포(melanocyte)는 모발색과 피부색을 결정하는 페놀류의 생체고분자 물질로 검은 단백질의 복합체인 멜라닌(melanin)을 합성하고 분비하여, 자외선에 의한 피부손상을 막아줄 뿐 아니라 독성 약물의 흡수 등 여러 중요한 작용을 수행한다. 그러나 멜라닌의 과잉 생산은 기미, 주근깨와 같은 피부 질환을 초래하며, 심한 경우 피부암의 유발과도 관련이 있는 것으로 보고되고 있다.The epidermis, located at the outermost part of the body, is arranged in the order of the basal layer, the stratum corneum, and the stratum corneum. Among the various cell types that make up it, melanocytes are phenolic biopolymers that determine hair color and skin color. They synthesize and secrete melanin, a complex of black proteins, and only prevent skin damage caused by UV rays. It also performs several important functions, including absorption of toxic drugs. However, excessive production of melanin causes skin diseases such as melasma and freckles, and in severe cases, it is reported to be related to the induction of skin cancer.
일반적으로 피부색은 멜라닌의 함량과 분포 등에 따라 결정되며, 멜라닌 세포에서 생성된 후 세포 외부로 방출되는 멜라노좀(melanosome)의 수와 분포에 관련이 있다. 멜라닌은 멜라닌 세포의 멜라노좀이라는 피부 세포의 소기관에서 만들어져 주위의 케라티노사이트(keratinocyte)라는 표피 세포로 이동한다. 멜라닌은 티로신을 기질로 하여 티로신을 산화시키는 티로시나아제에 의해 산화 과정을 거치면서 만들어진다. 피부의 과색소 침착은 피부의 염증 반응 이후 체내 호르몬 이상, 유전 질환, 자외선 조사 등 여러 요인에 의해 발생할 수 있는데 주된 요인은 멜라닌 색소 합성 이상 및 분포 이상에 의한 것이다.In general, skin color is determined according to the content and distribution of melanin, and is related to the number and distribution of melanosomes produced in melanocytes and released to the outside of the cells. Melanin is produced in the organelles of skin cells called melanosomes of melanocytes and moves to surrounding epidermal cells called keratinocytes. Melanin is produced through an oxidation process by tyrosinase, which oxidizes tyrosine using tyrosine as a substrate. Hyperpigmentation of the skin can be caused by several factors, such as hormonal abnormalities, genetic diseases, and ultraviolet irradiation after the inflammatory reaction of the skin. The main factors are due to abnormalities in the synthesis and distribution of melanin pigments.
미적 기능을 중시하는 관점에서 티로시나아제 효소의 활성과 발현을 조절함으로서 미백을 목적으로 하는 다양한 소재에 대한 연구가 이루어지고 있으며, 지금까지 알려진 것으로는 하이드로퀴논(hydroquinone), 아스코르브산(ascorbic acid), 코지산(kojic acid), 레티놀(retinole), 알부틴(arbutin) 등이 있다. 그러나 강력한 멜라닌 생합성 저해활성을 보이는 소재는 대부분 색소세포의 변성 또는 치사를 일으키고, 세포 본래의 기능을 손상시키는 등의 부작용을 발생시킨다는 문제가 있다. Research on various materials for the purpose of whitening by regulating the activity and expression of the tyrosinase enzyme from the viewpoint of emphasizing the aesthetic function is being conducted. Hydroquinone and ascorbic acid are known so far , kojic acid, retinol, and arbutin. However, most materials exhibiting strong melanin biosynthesis inhibitory activity have a problem in that they cause side effects such as degeneration or death of pigment cells and impairing the original function of cells.
한편, 생태계 교란종은 생태계에 교란을 일으키거나, 교란을 일으킬 우려가 있는 야생생물이다. 번식력이 강한 외래종은 자연생태계에 유입되면서 토종서식지를 잠식하여 생태계의 균형을 깨고 종의 다양성을 떨어뜨리는 등 심각한 문제를 일으킬 수 있다. 이에 생태계에 악영향을 미칠 우려가 있는 종에 대하여 정부차원에서 생태계 교란 야생생물을 법으로 규정·규제하고, 관리기구를 설치하거나 외래종 생태계영향평가제도를 실시하여 수입여부를 결정하고 있다.On the other hand, an ecosystem-disrupting species is a wildlife that disturbs or is likely to cause disturbance in the ecosystem. Exotic species with strong fertility can cause serious problems such as encroaching on native habitats as they enter the natural ecosystem, disrupting the balance of the ecosystem and reducing species diversity. Accordingly, the government regulates and regulates wildlife disturbing the ecosystem by law for species that are likely to adversely affect the ecosystem, establishes a management organization, or implements an ecosystem impact assessment system for exotic species to determine whether to import them.
생태적·경제적·산업적·공중보건적 피해를 끼치는 일부 외래종은 생물다양성 보전 및 이용에 관한 법률 제2조 제4항에 의거하여 생태계 교란 야생생물로 규정된다. 또한 외래생물에 해당하지 아니하는 생물 중 특정 지역에서 생태계의 균형을 교란하거나 교란할 우려가 있는 생물, 유전자 변형을 통하여 생산된 유전자변형 생물체 중 생태계의 균형을 교란하거나 교란할 우려가 있는 생물이 이에 속한다. 생물다양성 보전 및 이용에 관한 법률에 의해 지정된 생태계 교란 야생생물로는 포유류 1종(뉴트리아), 양서류·파충류 2종(황소개구리, 붉은귀거북속 전종), 어류 2종(파랑볼우럭, 큰입배스), 곤충류 1종(꽃매미), 식물 15종(돼지풀, 단풍잎돼지풀, 서양등골나물, 털물참새피, 물참새피, 도깨비가지, 애기수영, 가시박, 서양금혼초, 미국쑥부쟁이, 양미역취, 가시상추, 갯줄풀, 영국갯끈풀, 개망초)으로 총 21종이다.Some invasive species that cause ecological, economic, industrial, and public health damage are defined as ecosystem disturbance wildlife according to Article 2 (4) of the Act on the Conservation and Use of Biodiversity. Also, among organisms that do not fall under the category of alien organisms, organisms that disturb or are likely to disturb the balance of the ecosystem in a specific region, or organisms that are likely to disturb or disturb the balance of the ecosystem among genetically modified organisms produced through genetic modification belong Ecosystem disturbance wildlife designated by the Biodiversity Conservation and Utilization Act includes one mammal (nutria), two amphibians and reptiles (bullfrog, all species of the genus Red-eared tortoise), and two species of fish (blue sea bass, bigmouth bass). ), 1 species of insects (flower locust), 15 species of plants (pigweed, maple leaf ragweed, sagebrush, fern sparrow, water sparrow, goblin branch, baby swimming, thorn gourd, sagebrush, American mugwort, There are a total of 21 species of yangmi yeokchwi, spiny lettuce, chrysanthemum, British chrysanthemum, algae).
단풍잎돼지풀(Ambrosia trifida)은 북아메리카가 원산지인 귀화식물로 여름철에 피는 꽃의 꽃가루가 알레르기성 비염이나 피부염을 일으킨다. 확산 속도도 빨라 농작물에 피해를 준다. Maple leaf ragweed ( Ambrosia trifida ) is a naturalized plant native to North America, and pollen of flowers blooming in summer causes allergic rhinitis or dermatitis. It spreads quickly, causing damage to crops.
한편, 개망초(Annual fleabane)는 우리나라에서 가장 흔하게 볼 수 있는 귀화식물이다. 확산하는 속도가 빨라 제때 제초작업을 하지 않으면 밭농사에 심한 타격을 입힌다. On the other hand, annual fleabane is the most common naturalized plant in Korea. It spreads rapidly, and if weeding is not done in time, it will seriously damage the field farming.
이에 본 발명자들은 생태계 교란종으로 분류되어있는 단풍잎돼지풀 및 개망초를 기능성 원료로 활용하기 위하여 연구한 결과, 단풍잎돼지풀 및 개망초 발효 추출물의 현저한 피부 미백 및 주름 개선 효과를 확인함으로써 본 발명을 완성하게 되었다.Accordingly, the present inventors completed the present invention by confirming the remarkable skin whitening and wrinkle-improving effects of maple leaf ragweed and ragweed fermented extract as a result of research to utilize them as functional raw materials, which are classified as ecosystem disturbing species. became
따라서 본 발명의 목적은, 단풍잎돼지풀 및 개망초 발효 추출물을 유효성분으로 포함하는 피부 미백 또는 주름 개선용 화장료, 피부외용제 또는 식품 조성물을 제공하는 것이다.Accordingly, it is an object of the present invention to provide a cosmetic, external preparation for skin or food composition for skin whitening or wrinkle improvement, comprising a fermented extract of maple leaf ragweed and ragweed as an active ingredient.
상기 목적을 달성하기 위하여, 본 발명은 단풍잎돼지풀 및 개망초 발효 추출물을 유효성분으로 포함하는 피부 미백 또는 주름 개선용 화장료 조성물을 제공한다.In order to achieve the above object, the present invention provides a cosmetic composition for skin whitening or wrinkle improvement comprising a fermented extract of maple leaf ragweed and ragweed as an active ingredient.
또한, 본 발명은 단풍잎돼지풀 및 개망초 발효 추출물을 유효성분으로 포함하는 피부 미백 또는 주름 개선용 피부 외용제 조성물을 제공한다. In addition, the present invention provides a composition for external application for skin whitening or wrinkle improvement comprising a fermented extract of maple leaf ragweed and ragweed as an active ingredient.
또한, 단풍잎돼지풀 및 개망초 발효 추출물을 유효성분으로 포함하는 피부 미백 또는 주름 개선용 식품 조성물을 제공한다.In addition, there is provided a food composition for skin whitening or wrinkle improvement comprising a maple leaf ragweed and fermented extract of fermented ragweed as an active ingredient.
본 발명에 따른 단풍잎돼지풀 및 개망초 발효 추출물은 티로시나아제의 활성을 억제하고, 멜라닌 생성 세포에서 멜라닌 생성을 억제하는 효과를 가지고 있을 뿐만 아니라, 엘라스타아제의 저해 활성이 우수한바, 피부 미백 또는 주름 개선을 위한 화장료, 피부 외용제 및 식품 분야 등에서 다양하게 활용될 수 있다.The fermented maple leaf ragweed extract according to the present invention inhibits the activity of tyrosinase and has the effect of inhibiting the production of melanin in melanin-producing cells, and has excellent inhibitory activity of elastase, skin whitening or It can be used in various fields such as cosmetics for wrinkle improvement, external preparations for skin, and food.
도 1은 본 발명에 따른 단풍잎돼지풀 및 개망초 발효 추출물의 전자공여능을 측정한 결과를 나타낸 도이다.
도 2는 본 발명에 따른 단풍잎돼지풀 및 개망초 발효 추출물의 티로시나아제 저해 활성을 측정한 결과를 나타낸 도이다.
도 3은 본 발명에 따른 단풍잎돼지풀 및 개망초 발효 추출물의 엘라스타아제 저해 활성을 측정한 결과를 나타낸 도이다.
도 4는 MTT 분석을 통해 본 발명에 따른 단풍잎돼지풀 및 개망초 발효 추출물의 새포 독성을 측정한 결과를 나타낸 도이다.
도 5는 웨스턴 블롯팅을 통해 본 발명에 따른 단풍잎돼지풀 및 개망초 발효 추출물이 색소침착 관련 단백질의 억제 활성에 미치는 영향을 확인한 결과를 나타낸 도이다.
도 6은 RT-PCR을 통해 본 발명에 따른 단풍잎돼지풀 및 개망초 발효 추출물이 색소침착 관련 인자의 mRNA 발현에 미치는 영향을 확인한 결과를 나타낸 도이다.
도 7은 단풍잎돼지풀 추출물, 개망초 추출물과, 단풍잎돼지풀 및 개망초 발효 추출물의 전자공여능을 비교한 결과를 나타낸 도이다.
도 8은 단풍잎돼지풀 추출물, 개망초 추출물과, 단풍잎돼지풀 및 개망초 발효 추출물의 엘라스타아제 억제 활성을 비교한 결과를 나타낸 도이다.1 is a view showing the results of measuring the electron donating ability of maple leaf ragweed and fermented fermented extracts of Amanita fermented in accordance with the present invention.
Figure 2 is a diagram showing the results of measuring the tyrosinase inhibitory activity of the fermented extracts of maple leaf ragweed and A. ragweed according to the present invention.
Figure 3 is a diagram showing the result of measuring the elastase inhibitory activity of the fermented extract of maple leaf ragweed and A. ragweed according to the present invention.
Figure 4 is a diagram showing the results of measuring the cell toxicity of the fermented extract of maple leaf ragweed according to the present invention through MTT analysis.
5 is a view showing the results of confirming the effect of the fermented extract of Maple Leaf ragweed and Alfalfa root according to the present invention on the inhibitory activity of pigmentation-related proteins through Western blotting.
6 is a view showing the results of confirming the effect of the fermented extracts of maple leaf ragweed and ragweed according to the present invention on the mRNA expression of pigmentation-related factors through RT-PCR.
7 is a diagram showing the results of comparing the electron donating ability of a maple leaf ragweed extract, a ragweed extract, and a maple leaf ragweed extract and a fermented ragweed extract.
Figure 8 is a diagram showing the results of comparing the elastase inhibitory activity of the maple leaf ragweed extract, the ragweed extract and the fermented maple leaf ragweed extract.
이하, 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명의 일 양태에 따르면, 본 발명은 단풍잎돼지풀(Ambrosia trifida) 및 개망초(Annual fleabane) 발효 추출물을 유효성분으로 포함하는 피부 미백 또는 주름 개선용 화장료 조성물을 제공한다.According to one aspect of the present invention, the present invention provides a cosmetic composition for skin whitening or wrinkle improvement comprising a fermented extract of maple leaf ragweed ( Ambrosia trifida ) and annual fleabane as active ingredients.
본 발명에 있어서, “추출물”은 적절한 용매를 이용하여 단풍잎돼지풀 및 개망초의 발효물로부터 추출한 것이며, 예를 들어 단풍잎돼지풀 및 개망초의 발효물의 조추출물, 극성용매 가용 추출물 또는 비극성용매 가용 추출물을 모두 포함한다. 또한, 상기 추출물은 추출, 분획 또는 정제의 각 단계에서 얻어지는 모든 추출액, 분획물, 그들의 희석액, 농축액 또는 건조물을 모두 포함하는 개념이다.In the present invention, the "extract" is extracted from the fermented product of Maple Leaf ragweed and Alfalfa root using an appropriate solvent, for example, a crude extract, a polar solvent-soluble extract or a non-polar solvent-soluble extract include all In addition, the extract is a concept including all extracts, fractions, dilutions, concentrates or dried products obtained in each step of extraction, fractionation or purification.
본 발명의 조성물의 유효성분인 단풍잎돼지풀 및 개망초 발효 추출물은 통상적인 추출 방법에 의해 얻을 수 있고 또는 시판되는 것을 구입하여 사용할 수도 있다.The active ingredients of the composition of the present invention, the fermented extract of Maple Leaf ragweed and Alfalfa root, may be obtained by a conventional extraction method, or may be purchased and used commercially.
상기 단풍잎돼지풀 및 개망초 발효 추출물은 원료 식물의 다양한 부위로부터 추출 또는 분획될 수 있으며, 바람직하게는 단풍잎돼지풀 및 개망초의 잎, 줄기 및 뿌리로부터 추출되는 것이다.The fermented maple leaf ragweed and fermented extract may be extracted or fractionated from various parts of the raw material plant, and is preferably extracted from the leaves, stems and roots of maple leaf ragweed and ragweed.
상기 단풍잎돼지풀 및 개망초 발효 추출물은 당업계에 공지된 추출, 분리 및 분획하는 방법을 사용하여 천연으로부터 추출, 분리 및 분획하여 수득한 것을 사용할 수 있다. The fermented maple leaf ragweed extract and fermented ragweed extract may be obtained by extraction, separation and fractionation from nature using methods known in the art for extraction, separation and fractionation.
본 발명의 일 실시예에서, 단풍잎돼지풀 및 개망초 발효 추출물은 단풍잎돼지풀 및 개망초를 혼합한 후, 온양온천수를 첨가하여 발효시킨 후 추출하여 제조하였다. In one embodiment of the present invention, the fermented extract of maple leaf ragweed and ragweed was prepared by mixing maple leaf ragweed and ragweed, and then fermenting it by adding Onyang hot spring water, followed by extraction.
상기 단풍잎돼지풀 및 개망초는 바람직하게는 동량, 즉 1:1의 중량비율로 혼합할 수 있다. The maple leaf ragweed and ragweed may be mixed in the same amount, that is, in a weight ratio of 1:1.
본 발명의 일 실시예에서, 상기 온양온천수는 pH 7 내지 9, 바람직하게는 pH 8 내지 9인 것이 바람직하나, 이에 제한되지 않는다.In one embodiment of the present invention, the hot spring water preferably has a pH of 7 to 9, preferably a pH of 8 to 9, but is not limited thereto.
본 발명의 일 실시예에서, 상기 온양온천수는 칼륨 1 내지 2mg/L, 나트륨 40 내지 50mg/L, 칼슘 5 내지 10mg/L, 실리콘 20 내지 30mg/L, 리튬 0.05 내지 0.20mg/L, 스트론튬 0.1 내지 0.5mg/L 및 마그네슘 0.1 내지 0.4mg/L을 포함하는 것이 바람직하며, 더 바람직하게는 칼륨 1.59mg/L, 나트륨 45.5mg/L, 칼슘 6.75mg/L, 실리콘 25.85mg/L, 리튬 0.12mg/L, 스트론튬 0.27mg/L 및 마그네슘 0.21mg/L을 포함하나, 이에 제한되지 않는다.In an embodiment of the present invention, the hot spring water is
본 발명에 있어서, 발효란 유기물이 미생물작용에 의해 분해 및 변화하여 유용한 물질이 생산되는 것을 의미한다. 본 발명에서 발효를 위해 사용될 수 있는 것은 발효를 통해 단풍잎돼지풀 및 개망초의 활성을 증진시킬 수 있는 미생물이라면 그 종류가 제한되지 않으나, 바람직하게는 락토바실러스 속 균주이다. 상기 락토바실러스 속 균주에는 예를 들어, 락토바실러스 람노서스(Lactobacillus rhamnosus), 락토바실러스 파라카세이(Lactobacillus paracasei ), 락토바실러스 카제이(Lactobacillus casei), 락토바실러스 플란타룸(Lactobacillus plantarum), 락토바실러스 불가리쿠스(Lactobacillus bulgaricus), 락토바실러스 락티스(Lactobacillus lactis), 락토바실러스 퍼멘텀(Lactobacillus fermentum) 등이 포함되나 이에 제한되지 않는다. 본 발명의 구체예에서는, 발효를 위한 미생물로 락토바실러스 락토바실러스 람노서스(Lactobacillus rhamnosus) 및 락토바실러스 파라카세이(Lactobacillus paracasei )를 혼합하여 사용하였으나, 이에 제한되지 않는다.In the present invention, fermentation means that organic matter is decomposed and changed by microbial action to produce useful substances. The type of microorganism that can be used for fermentation in the present invention is not limited, as long as it is a microorganism capable of enhancing the activity of maple leaf ragweed and aloe vera through fermentation, but is preferably a Lactobacillus sp. The Lactobacillus sp include, for example, Lactobacillus ramno suspension (Lactobacillus rhamnosus), Lactobacillus para Kasei (Lactobacillus paracasei), Lactobacillus casei (Lactobacillus casei), Lactobacillus Planta room (Lactobacillus plantarum), Lactobacillus Bulgaria Cus ( Lactobacillus bulgaricus ), Lactobacillus lactis ( Lactobacillus lactis ), Lactobacillus fermentum ( Lactobacillus fermentum ), and the like are included, but are not limited thereto. In exemplary embodiments of the present invention, but used as a microorganism for fermentation mixture of Lactobacillus Lactobacillus ramno suspension (Lactobacillus rhamnosus), and Lactobacillus para Kasei (Lactobacillus paracasei), but is not limited thereto.
본 발명에서 발효는 25 내지 40℃에서 1일 내지 5일간 수행하는 것일 수 있으며, 바람직하게는 32 내지 40℃에서 2일 내지 4일간 수행하는 것일 수 있으나, 이에 제한되지 않는다. 본 발명의 일 실시예에서는 락토바실러스 속 균주를 접종한 후, 37℃에서 3일간 발효시켜 본 발명에 따른 단풍잎돼지풀 및 개망초 발효물을 제조하였다.Fermentation in the present invention may be carried out at 25 to 40 ℃ for 1 to 5 days, preferably it may be carried out at 32 to 40 ℃ for 2 to 4 days, but is not limited thereto. In one embodiment of the present invention, after inoculating the Lactobacillus sp. strain, it was fermented at 37° C. for 3 days to prepare a fermented maple leaf ragweed according to the present invention.
상기 단풍잎돼지풀 및 개망초 발효 추출물을 수득하기 위한 적절한 용매로는 물 또는 유기용매를 사용할 수 있으며, 약학적으로 허용되는 유기용매라면 어느 것을 사용해도 무방하다. 예를 들어, 상기 용매로는 물, 메탄올(methanol), 에탄올(ethanol), 프로판올(propanol), 이소프로판올(isopropanol), 부탄올(butanol) 등의 탄소수 1 내지 4의 알코올 등을 단독으로 또는 2종 이상 혼합하여 사용할 수 있으며, 물을 사용하는 것이 가장 바람직하다.Water or an organic solvent may be used as a suitable solvent for obtaining the fermented extract of maple leaf ragweed and fermented ragweed, and any pharmaceutically acceptable organic solvent may be used. For example, as the solvent, an alcohol having 1 to 4 carbon atoms, such as water, methanol, ethanol, propanol, isopropanol, butanol, etc. alone or two or more It can be used by mixing, and it is most preferable to use water.
상기 추출 온도는 50 내지 100℃인 것이 바람직하다. 또한, 추출방법으로는 열수추출법, 냉침추출법, 환류냉각추출법, 용매추출법, 수증기증류법, 초음파추출법, 용출법, 압착법 등의 다양한 방법이 사용될 수 있으나 이에 제한되지는 않고, 바람직하게는 열수추출법 또는 환류냉각추출법을 사용하는 것이다.The extraction temperature is preferably 50 to 100 ℃. In addition, as the extraction method, various methods such as hot water extraction method, cold extraction method, reflux cooling extraction method, solvent extraction method, steam distillation method, ultrasonic extraction method, elution method, compression method, etc. may be used, but it is not limited thereto, and preferably hot water extraction method or Reflux cooling extraction method is used.
상기 단풍잎돼지풀 및 개망초 발효 추출물은 당업계에서 알려진 통상의 방법으로 상온에서 냉침, 가열 및 여과하여 액상물을 얻을 수 있으며, 추가로 용매를 증발, 분무건조 또는 동결건조 할 수도 있다. 또한, 상기 단풍잎돼지풀 및 개망초 발효 추출물은 감압 증류 및 동결 건조 또는 분무 건조 등과 같은 추가적인 과정에 의해 분말 상태로 제조할 수도 있다. 또한, 상기 추출물을 실리카겔 컬럼 크로마토그래피(silica gel column chromatography), 박층크로마토그래피(thin layer chromatography), 고성능 액체 크로마토그래피(high performance liquid chromatography) 등과 같은 다양한 크로마토그래피를 이용하여 추가로 정제된 분획으로도 얻을 수 있다.The fermented maple leaf ragweed extract and fermented ragweed extract can be obtained by cooling, heating and filtering at room temperature by a conventional method known in the art, and additionally evaporating the solvent, spray-drying or freeze-drying. In addition, the fermented extracts of maple leaf ragweed and ragweed may be prepared in a powder state by an additional process such as distillation under reduced pressure and freeze-drying or spray-drying. In addition, the extract is further purified by using various chromatography methods such as silica gel column chromatography, thin layer chromatography, high performance liquid chromatography, etc. can be obtained
따라서 본 발명에서 사용되는 상기 단풍잎돼지풀 및 개망초 발효 추출물은 추출, 분획 또는 정제의 각 단계에서 얻어지는 모든 추출물, 분획물 및 정제물, 그들의 희석액, 농축액 또는 건조물을 모두 포함하는 개념이다.Therefore, the fermented maple leaf ragweed extract used in the present invention is a concept including all extracts, fractions and purified products obtained in each step of extraction, fractionation or purification, and their dilutions, concentrates or dried products.
본 발명의 일 실시예에 따르면, 단풍잎돼지풀 및 개망초 발효 추출물은 티로시나아제 효소 실험에서 티로시나아제 활성을 억제시키며, 멜라닌 축적이 유도된 세포 모델에서 멜라닌 생성을 억제하는 우수한 효과를 가지고 있다. 따라서, 본 발명의 단풍잎돼지풀 및 개망초 발효 추출물은 피부 미백을 위한 화장료 조성물로 유용하게 사용될 수 있다. 또한, 본 발명의 다른 실시예에 따르면, 단풍잎돼지풀 및 개망초 발효 추출물은 엘라스타아제의 활성을 억제하는 우수한 효과를 가지고 있다. 따라서, 본 발명의 단풍잎돼지풀 및 개망초 발효 추출물은 주름 개선을 위한 화장료 조성물로 유용하게 사용될 수 있다.According to an embodiment of the present invention, the fermented extract of Maple Leaf ragweed and fermented ragweed inhibits tyrosinase activity in a tyrosinase enzyme experiment, and has an excellent effect of inhibiting melanin production in a cell model in which melanin accumulation is induced. Therefore, the fermented extracts of Maple Leaf ragweed and Alfalfa root of the present invention can be usefully used as a cosmetic composition for skin whitening. In addition, according to another embodiment of the present invention, the fermented extract of maple leaf ragweed and aloe vera has an excellent effect of inhibiting the activity of elastase. Therefore, the fermented extract of Maple leaf ragweed and Aerica larvae of the present invention can be usefully used as a cosmetic composition for wrinkle improvement.
상기 단풍잎돼지풀 및 개망초 발효 추출물은 화장료 조성물 총 중량에 대하여 0.01 내지 95중량%로 포함되는 것이 바람직하고, 더욱 바람직하게는 1 내지 80중량%로 포함될 수 있다. 그 함량이 0.01중량% 미만일 경우에는 피부 미백 또는 주름 개선 효과가 미미할 수 있으며, 95중량%를 초과할 경우에는 사용량 대비 효과 상승률이 낮아 비경제적일 수 있다.The fermented extract of maple leaf ragweed and fermented ragweed may be included in an amount of 0.01 to 95% by weight, more preferably 1 to 80% by weight, based on the total weight of the cosmetic composition. If the content is less than 0.01% by weight, the effect of skin whitening or wrinkle improvement may be insignificant, and if it exceeds 95% by weight, the effect increase rate is low compared to the amount used, which may be uneconomical.
본 발명의 화장료 조성물은 상기 유효성분 이외에 화장료 조성물에 통상적으로 사용되는 항산화제, 안정화제, 용해화제, 비타민, 안료, 향료 등과 같은 통상적인 보조제 및 담체가 더 포함될 수 있다. 예를 들어, 상기 화장료 조성물에는 글리세린, 부틸렌 글라이콜, 폴리옥시에칠렌 경화피마자유, 토코페릴 아세테이트, 시트릭산, 판테놀, 스쿠알란, 소듐 시트레이트, 알란토인 등의 보조성분이 추가로 더 포함될 수 있다.The cosmetic composition of the present invention may further include conventional adjuvants and carriers such as antioxidants, stabilizers, solubilizers, vitamins, pigments, fragrances, and the like, which are commonly used in cosmetic compositions in addition to the active ingredients. For example, the cosmetic composition may further include auxiliary components such as glycerin, butylene glycol, polyoxyethylene hydrogenated castor oil, tocopheryl acetate, citric acid, panthenol, squalane, sodium citrate, and allantoin. .
본 발명의 화장료 조성물은 기본적으로 피부에 도포되는 것이므로, 당업계의 화장료 조성물을 참조하여 통상적으로 제조되는 어떠한 제형으로도 제조될 수 있다. 예를 들어, 용액, 현탁액, 유탁액, 페이스트, 겔, 크림, 로션, 파우더, 비누, 계면활성제-함유 클린싱, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션 및 스프레이 등으로 제형화될 수 있으나, 이에 한정되는 것은 아니다. 보다 상세하게는, 유연 화장수, 영양 화장수, 영양크림, 마사지크림, 에센스, 아이크림, 클렌징크림, 클렌징폼, 클렌징워터, 마스크팩, 스프레이 또는 파우더의 제형으로 제조될 수 있다.Since the cosmetic composition of the present invention is basically applied to the skin, it may be prepared in any formulation conventionally prepared with reference to a cosmetic composition in the art. For example, it may be formulated as a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray, etc. The present invention is not limited thereto. More specifically, it may be prepared in the form of a flexible lotion, a nourishing lotion, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a mask pack, a spray or a powder.
본 발명의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로 동물성유, 식물성유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크, 산화아연 등이 포함될 수 있다.When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. may be included as carrier components. have.
본 발명의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로 락토스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트, 폴리아미드 파우더 등이 포함될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄, 디메틸 에테르 등의 추진체를 포함할 수 있다.When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, etc. may be included as carrier components, and in particular, in the case of a spray, additional chlorofluorohydrocarbon, propane / May contain propellants such as butane and dimethyl ether.
본 발명의 제형이 용액 또는 유탁액인 경우에는 담체 성분으로 용매, 용해화제, 유탁화제 등이 포함될 수 있고, 구체적으로 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜, 소르비탄의 지방산 에스테르 등이 포함될 수 있다.When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer, emulsifier, etc. may be included as carrier components, and specifically, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene Glycol, 1,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid ester of sorbitan, and the like may be included.
본 발명의 제형이 현탁액인 경우에는 담체 성분으로 물, 에탄올, 프로필렌글리콜 등의 액상 희석제; 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르, 폴리옥시에틸렌 소르비탄 에스테르 등의 현탁제; 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가, 트라칸트 등이 포함될 수 있다.When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol, propylene glycol; suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester; Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, tracanth, and the like may be included.
본 발명의 제형이 계면-활성제 함유 클린징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르 설페이트, 설포숙신산 모노에스테르, 이세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사르코시네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산 디에탄올아미드, 식물성유, 라놀린유도체, 에톡실화 글리세롤 지방산 에스테르 등이 포함될 수 있다.When the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide as carrier components ether sulfates, alkylamidobetaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, ethoxylated glycerol fatty acid esters, and the like may be included.
본 발명의 다른 양태에 따르면, 본 발명은 단풍잎돼지풀 및 개망초 발효 추출물을 유효성분으로 포함하는 피부 미백 또는 주름 개선용 피부 외용제 조성물을 제공한다. According to another aspect of the present invention, the present invention provides a composition for external application for skin for skin whitening or wrinkle improvement comprising a fermented extract of maple leaf ragweed and fermented ragweed as active ingredients.
본 발명의 조성물이 피부 외용제 조성물로 사용되는 경우, 추가로 지방 물질, 유기 용매, 용해제, 농축제 및 겔화제, 연화제, 항산화제, 현탁화제, 안정화제, 발포제(foaming agent), 방향제, 계면활성제, 물, 이온형 또는 비이온형 유화제, 충전제, 금속이온봉쇄제 및 킬레이트화제, 보존제, 비타민, 차단제, 습윤화제, 필수 오일, 염료, 안료, 친수성 또는 친유성 활성제, 지질 소낭 또는 피부용 외용제에 통상적으로 사용되는 임의의 다른 성분과 같은 피부 과학 분야에서 통상적으로 사용되는 보조제를 함유할 수 있다. 또한 상기 성분들은 피부 과학분야에서 일반적으로 사용되는 양으로 도입될 수 있다. When the composition of the present invention is used as a composition for external application to the skin, additionally, a fatty substance, an organic solvent, a solubilizer, a thickening agent and a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, a surfactant , water, ionic or non-ionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or common for external applications for the skin It may contain adjuvants commonly used in the field of dermatology, such as any other ingredients used as a dermatome. In addition, the above ingredients may be introduced in an amount generally used in the field of dermatology.
본 발명의 조성물이 피부 외용제 제형으로 제공될 경우, 이에 제한되는 것은 아니나, 연고, 패취, 겔, 크림 또는 분무제와 같은 제형을 가질 수 있다.When the composition of the present invention is provided as a formulation for external application to the skin, the present invention is not limited thereto, but may have formulations such as ointments, patches, gels, creams or sprays.
본 발명의 또 다른 양태에 따르면, 본 발명은 단풍잎돼지풀 및 개망초 발효 추출물을 유효성분으로 포함하는 피부 미백 또는 주름 개선용 식품 조성물을 제공한다.According to another aspect of the present invention, the present invention provides a food composition for skin whitening or wrinkle improvement, comprising the fermented extract of maple leaf ragweed and fermented ragweed as an active ingredient.
본 발명의 조성물이 식품 조성물로 이용되는 경우, 본 발명의 식품 조성물은 피부 미백 효과를 가지는 식품을 의미하는 것으로, 장기적으로 복용하였을 때 인체에 무해해야 한다.When the composition of the present invention is used as a food composition, the food composition of the present invention means a food having a skin whitening effect, and should be harmless to the human body when taken for a long time.
상기 식품의 종류에는 특별한 제한은 없다. 상기 물질을 첨가할 수 있는 식품의 예로는 육류, 소시지, 빵, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 수프, 음료수, 차, 드링크제, 알코올 음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강식품을 모두 포함한다.There is no particular limitation on the type of the food. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, There are alcoholic beverages, vitamin complexes, and the like, and includes all health foods in the ordinary sense.
본 발명의 실시예에서, 본 발명의 식품 조성물은 식품 첨가물일 수 있다. 상기 식품 첨가물은 상기 단풍잎돼지풀 및 개망초 발효 추출물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효성분의 혼합양은 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다.In an embodiment of the present invention, the food composition of the present invention may be a food additive. The food additive may be added as it is, or used with other foods or food ingredients, and may be appropriately used according to a conventional method. The mixed amount of the active ingredient may be appropriately determined according to the purpose of use (prevention, health or therapeutic treatment).
본 발명의 실시예에서, 본 발명의 식품 조성물은 건강음료 조성물일 수 있다. 상기 건강음료 조성물은 단풍잎돼지풀 및 개망초 발효 추출물 외에 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 포함할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토오스, 수크로오스와 같은 디사카라이드, 및 덱스트린, 사이클로덱스트린과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100 ml 당 일반적으로 약 0.01 내지 10 g, 바람직하게는 약 0.01 내지 0.1 g 이다.In an embodiment of the present invention, the food composition of the present invention may be a health drink composition. The health beverage composition may include various flavoring agents or natural carbohydrates as additional components, such as a conventional beverage, in addition to the fermented extract of Maple Leaf ragweed and Alfalfa root. As the above-mentioned natural carbohydrates, monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharin and aspartame may be used. The proportion of the natural carbohydrate is generally about 0.01 to 10 g, preferably about 0.01 to 0.1 g per 100 ml of the composition of the present invention.
상기 외에 본 발명의 조성물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산 음료에 사용되는 탄산화제 등을 포함할 수 있다. 그 밖에 본 발명의 조성물은 천연 과일주스, 과일주스 음료 및 야채 음료의 제조를 위한 과육을 포함할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 크게 중요하진 않지만 본 발명의 조성물 100 중량부 당 0.01 내지 0.1 중량부의 범위에서 선택되는 것이 일반적이다.In addition to the above, the composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, Carbonating agents used in carbonated beverages, etc. may be included. In addition, the composition of the present invention may contain the pulp for the production of natural fruit juice, fruit juice beverage, and vegetable beverage. These components may be used independently or in combination. The proportion of these additives is not very important, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as being limited by these examples.
실시예Example 1. One. 단풍잎돼지풀Maple Leaf Pig Grass 및 개망초 발효 추출물 제조 and Manufacture of fermented extract
단풍잎돼지풀 및 개망초 발효 추출물을 제조하기 위하여, 단풍잎돼지풀과 개망초를 각각 1 내지 3cm로 잘라 동량씩 혼합한 후 1,300 메쉬로 분쇄하였다. 분쇄된 단풍잎돼지풀 및 개망초 혼합물을 127℃에서 30분 동안 멸균하였다. 상기 혼합물을 발효시키기 위해, 온양온천수를 1:1(w/v)로 혼합한 후 균질화시켜 온천수 혼합물을 제조하였다. 상기 온양온천수의 성분은 표 1에 나타내었다.In order to prepare a fermented extract of maple leaf ragweed and ragweed, the maple leaf ragweed and algae were cut into 1 to 3 cm, respectively, and mixed in equal amounts, and then pulverized to 1,300 mesh. A mixture of crushed maple leaf ragweed and ragweed was sterilized at 127° C. for 30 minutes. In order to ferment the mixture, hot spring water was mixed at a ratio of 1:1 (w/v) and then homogenized to prepare a hot spring water mixture. The components of the hot spring water are shown in Table 1.
상기 온천수 혼합물 총 중량에 대하여 수크로오스 3중량%, 효모 추출물 2중량% 및 디포타슘 포스페이트 0.2중량%를 더 첨가하였다. 다음으로, 상기 과정을 통해 준비된 온천수 혼합물에 고농도로 활성화된 유산균인 락토바실러스 속 균주를 10%의 농도로 접종하였으며, 이를 37℃, 120rpm 조건에서 72시간 동안 발효시켰다. 락토바실러스 속 균주는 시판되는 것을 이용하였으며, 구체적으로, 락토바실러스 람노서스(Lactobacillus rhamnosus) 및 락토바실러스 파라카세이(Lactobacillus paracasei)가 혼합된 것이다. 3% by weight of sucrose, 2% by weight of yeast extract and 0.2% by weight of dipotassium phosphate were further added based on the total weight of the hot spring water mixture. Next, the Lactobacillus sp. strain, which is a highly activated lactic acid bacterium, was inoculated at a concentration of 10% in the hot spring water mixture prepared through the above process, and this was fermented for 72 hours at 37°C and 120rpm conditions. Lactobacillus sp. strains were used commercially, specifically, Lactobacillus rhamnosus ( Lactobacillus rhamnosus ) and Lactobacillus paracasei ( Lactobacillus paracasei ) is a mixture.
발효 종료 후, 단풍잎돼지풀 및 개망초 발효물을 수직 환류 냉각기가 부착된 추출 플라스크에 옮기고, 시료 중량의 10배수 증류수를 용매로 첨가한 후, 100℃에서 60분 동안 추출을 3회 반복하여 열수 추출하여, 단풍잎돼지풀 및 개망초 발효 추출물을 제조하였다. After the fermentation is complete, the fermented maple leaf ragweed and fermented ragweed are transferred to an extraction flask with a vertical reflux cooler, distilled
실시예 2. 단풍잎돼지풀 및 개망초 발효 추출물의 전자공여능 측정Example 2. Measurement of electron donating ability of maple leaf ragweed and fermented extract
전자공여능(EDA: electron donating abilities) 측정을 위해, DPPH(1,1-diphenyl-2-picrylhydrazyl) 60μl와 농도별 단풍잎돼지풀 및 개망초 발효 추출물을 120μl씩 넣고 혼합한 후 15분간 방치한 다음 마이크로플레이트 리더를 이용하여 517nm에서 흡광도를 측정하였다. 전자공여능은 하기 수학식 1과 같이 계산하였으며, 그 결과를 도 1에 나타내었다.To measure electron donating abilities (EDA), 60 μl of DPPH (1,1-diphenyl-2-picrylhydrazyl) and 120 μl of fermented extracts of maple leaf ragweed and fermented ragweed per concentration were mixed, left for 15 minutes, and then microplate Absorbance was measured at 517 nm using a reader. The electron donating ability was calculated as in
[수학식 1][Equation 1]
전자공여능(%) =(1 - 시료첨가군의 흡광도/무첨가군의 흡광도)×100Electron donating ability (%) = (1 - absorbance of sample added group / absorbance of no addition group) × 100
도 1에 나타낸 바와 같이, 단풍잎돼지풀 및 개망초 발효 추출물의 농도가 증가함에 따라 전자공여능이 증가하는 효과를 확인하였다. As shown in FIG. 1 , the effect of increasing the electron donating ability was confirmed as the concentration of the fermented extracts of Maple Leaf ragweed and fermented ragweed increased.
실시예Example 3. 3. 단풍잎돼지풀Maple Leaf Pig Grass 및 개망초 발효 추출물의 티로시나아제 저해 활성 측정 and tyrosinase inhibitory activity measurement of fermented extract
미백 효과를 확인하기 위한 티로시나아제 저해 활성 측정은 다음과 같이 측정하였다. 67mM 인산염완충액(pH 6.8) 80μl에 10mM L-DOPA(Sigma, USA)를 녹인 기질액 40μl 및 시료 용액(단풍잎돼지풀 및 개망초 발효 추출물) 40μl의 혼합액을 제조한 후, 200U/ml 버섯 티로시나아제(Sigma, USA) 40μl를 첨가하여 37℃에서 10분간 반응시켰다. 이 후 반응액 중에 생성된 DOPA 크롬을 확인하기 위해 492nm에서 흡광도를 측정하였다. 티로시나아제 저해활성은 하기 수학식 2와 같이 계산하였으며, 그 결과를 도 2에 나타내었다.Tyrosinase inhibitory activity to confirm the whitening effect was measured as follows. After preparing a mixed solution of 40 μl of substrate solution and 40 μl of sample solution (fermented extract of Maple leaf ragweed and Astragalus fermented extract) in which 10 mM L-DOPA (Sigma, USA) was dissolved in 80 μl of 67 mM phosphate buffer (pH 6.8), 200 U/ml mushroom tyrosinase (Sigma, USA) 40 μl was added and reacted at 37° C. for 10 minutes. Thereafter, absorbance was measured at 492 nm to confirm the DOPA chromium generated in the reaction solution. Tyrosinase inhibitory activity was calculated as in
[수학식 2][Equation 2]
저해율(%) =(1 - 시료첨가군의 흡광도/무첨가군의 흡광도)×100Inhibition rate (%) = (1 - absorbance of sample added group / absorbance of no addition group) × 100
도 2에 나타낸 바와 같이, 단풍잎돼지풀 및 개망초 발효 추출물의 농도가 증가함에 따라 티로시나아제 저해 활성이 증가하는 등 미백 효과가 우수함을 확인하였다. As shown in FIG. 2 , it was confirmed that the whitening effect was excellent, such as the increase in the tyrosinase inhibitory activity as the concentration of the fermented extract of Maple Leaf ragweed and Algae fermented extract increased.
실시예 4. 단풍잎돼지풀 및 개망초 발효 추출물의 엘라스타아제 저해 활성 측정Example 4. Elastase inhibitory activity measurement of maple leaf ragweed and fermented extract
주름 개선 효과를 확인하기 위하여 피부 노화를 촉진하는 대표적 효소인 엘리스타아제 저해 활성을 확인하였다. 구체적으로, 시료 용액(단풍잎돼지풀 및 개망초 발효 추출물)을 일정 농도가 되도록 조제하여 40μl씩 96-웰 플레이트에 분주하고, 50 mM tris-HCl 완충액(pH8.6)에 녹인 2.5 U/ml 돼지 췌장 엘리스타아제(Sigma, USA) 용액 40μl를 가한 후 기질로 50mM tris-HCl 완충액(pH 8.6)에 녹인 N-succinyl-(L-Ala)3-p-nitroanilide(0.5 mg/ml)을 80μl를 첨가하였다. 이를 37℃에서 30분간 반응시켰으며, 기질로부터 생성되는 p-nitroanilide의 생성량을 확인하기 위해 445nm에서 흡광도를 측정하였다. 엘리스타아제 저해활성은 하기 수학식 3과 같이 계산하였으며, 그 결과를 도 3에 나타내었다.In order to confirm the wrinkle improvement effect, the inhibitory activity of elastase, a representative enzyme that promotes skin aging, was confirmed. Specifically, a sample solution (a fermented extract of maple leaf ragweed and fermented ragweed) was prepared to a certain concentration, and 40 μl of each was dispensed into a 96-well plate, and 2.5 U/ml of porcine pancreas dissolved in 50 mM tris-HCl buffer (pH8.6). After adding 40 μl of elastase (Sigma, USA) solution, 80 μl of N-succinyl-(L-Ala)3-p-nitroanilide (0.5 mg/ml) dissolved in 50 mM tris-HCl buffer (pH 8.6) as a substrate was added. did. This was reacted at 37° C. for 30 minutes, and absorbance was measured at 445 nm to confirm the amount of p-nitroanilide produced from the substrate. Elastase inhibitory activity was calculated as in Equation 3 below, and the results are shown in FIG. 3 .
[수학식 3][Equation 3]
저해율(%) =(1 - 시료첨가군의 흡광도/무첨가군의 흡광도)×100Inhibition rate (%) = (1 - absorbance of sample added group / absorbance of no addition group) × 100
도 3에 나타낸 바와 같이, 단풍잎돼지풀 및 개망초 발효 추출물의 농도가 증가함에 따라 엘리스타아제 저해 활성이 증가하였으며, 이를 통해 상기 발효 추출물의 주름 개선 효과가 우수한 것을 확인하였다. As shown in FIG. 3 , as the concentration of the fermented extracts of Maple Leaf ragweed and A. fermented extract increased, the elastase inhibitory activity increased, and it was confirmed that the wrinkle improvement effect of the fermented extract was excellent.
실시예Example 5. 5. 단풍잎돼지풀Maple Leaf Pig Grass 및 개망초 발효 추출물의 세포 독성 측정 and cytotoxicity measurement of fermented extract
본 실험에 이용한 흑색종 세포인 B16F10은 ATCC(USA)에서 구입하였다. B16F10 세포 배양은 10% FBS와 1% 페니실린/스트렙토마이신(100 U/ml)을 첨가한 DMEM 배지를 사용하였으며, 37℃, 5% CO2 배양기에 적응시켜 계대 배양하였다. B16F10, a melanoma cell used in this experiment, was purchased from ATCC (USA). For B16F10 cell culture, DMEM medium supplemented with 10% FBS and 1% penicillin/streptomycin (100 U/ml) was used, and subcultured at 37° C. and 5% CO 2 adapted to an incubator.
세포 생존율 측정을 위해, B16F10 세포를 96-웰 플레이트에 1×105cells/well이 되도록 180μl씩 분주하였고, 시료(단풍잎돼지풀 및 개망초 발효 추출물)를 농도별로 조제하여 20μl씩 첨가한 후 37℃, 5% CO2 배양기에서 24시간 배양하였다. 여기에 2.5mg/ml 농도로 제조한 MTT 용액을 40μl씩 첨가하여 4시간 동안 배양하였다. 이 후 배양액을 제거하고 각 웰당 DMSO 100μl를 가하여 실온에서 10분간 반응시킨 뒤 ELISA 리더로 540nm에서 흡광도를 측정하였다. 세포 생존율 측정은 하기 수학식 4와 같이 계산하였으며, 그 결과를 도 4에 나타내었다. For cell viability measurement, 180 μl of B16F10 cells were aliquoted in a 96-well plate to 1×10 5 cells/well, and samples (maple leaf ragweed and fermented fermented fermented extracts) were prepared by concentration and 20 μl was added at 37°C. , 5% CO 2 Incubated for 24 hours in an incubator. Here, 40 μl of MTT solution prepared at a concentration of 2.5 mg/ml was added and incubated for 4 hours. Thereafter, the culture medium was removed, and 100 μl of DMSO was added to each well, followed by reaction at room temperature for 10 minutes, and absorbance was measured at 540 nm with an ELISA reader. Cell viability was calculated as in Equation 4 below, and the results are shown in FIG. 4 .
[수학식 4][Equation 4]
세포생존율(%) =(1 - 시료첨가군의 흡광도/무첨가군의 흡광도)×100Cell viability (%) = (1 - Absorbance of the sample added group / Absorbance of the non-added group) × 100
도 4에 나타낸 바와 같이, 단풍잎돼지풀 및 개망초 발효 추출물은 500μg/ml의 고 농도에서까지 약 97%의 세포 생존율을 나타내는 등 세포 독성이 없음을 확인하였다. As shown in FIG. 4 , it was confirmed that the fermented extracts of Maple leaf ragweed and A. fermented ragweed exhibited no cytotoxicity, such as showing a cell viability of about 97% even at a high concentration of 500 μg/ml.
실시예 6. 단풍잎돼지풀 및 개망초 발효 추출물의 색소침착 관련 인자 억제 활성 확인Example 6. Confirmation of pigmentation-related factor inhibitory activity of maple leaf ragweed and fermented extract
6-1. 색소침착 관련 인자 단백질의 억제 활성 확인6-1. Confirmation of inhibitory activity of pigmentation-related factor protein
MITF, TRP-1, TRP-2, 티로시나아제 등 색소침착 및 미백과 관련된 인자들의 활성을 단백질 수준에서 알아보기 위하여, B16F10 세포에 단풍잎돼지풀 및 개망초 발효 추출물을 25, 50, 100μg/ml로 농도별로 처리하였다. 대조군은 α-MSH를 처리하여 멜라닌을 과발현한 구간으로 설정하였고, 정상군은 α-MSH를 처리하지 않은 구간으로 설정하였다. 이때 세포의 여러 조건에서도 그 발현 정도의 차이가 거의 없는 하우스키핑 유전자인 β-actin의 발현량을 확인하여 정량화하였다. To investigate the activity of factors related to pigmentation and whitening, such as MITF, TRP-1, TRP-2, and tyrosinase, at the protein level, 25, 50, and 100 μg/ml Treated by concentration. The control group was treated with α-MSH and set to the section in which melanin was overexpressed, and the normal group was set to the section not treated with α-MSH. At this time, the expression level of β-actin, a housekeeping gene with little difference in expression level, was confirmed and quantified under various cell conditions.
구체적으로, B16F10 세포를 100mm 티슈 컬쳐 디쉬에 1×106cells/well로 분주한 후 24시간 동안 배양하여 세포를 안정화시켰다. 배지를 제거한 후 α-MSH(100nm)를 2시간동안 처리한 후 각 발효 추출물을 농도 별로 처리한 배지로 교체하고 24시간동안 배양한 후 다시 배지를 제거하고 1× PBS로 2번 세척해주었다. 각 세포를 컴플리트 미니 1탭을 가한 RIPA 버퍼 10ml에 용해하여 4℃, 13,200rpm에서 20분간 원심분리 하였다. 원심분리하여 얻은 상층액을 BCA 단백질 분석 키트로 정량하였으며, 20μl의 단백질을 10% 아크릴아마이드 겔에서 전기영동하여 분리하였다. 분리된 단백질을 트랜스퍼 기기를 이용하여 PVDF(polyvinylidene fluoride) 멤브레인에 옮긴 다음, 실온에서 1시간동안 블로킹 완충액(5% skim milk in TBST)에서 배양시켰다. 1차 항체를 희석하여 4℃에서 밤새도록 배양한 다음, 다시 10분 간격으로 TBST(tris-buffered saline and tween 20)로 3회 세척하였다. 2차 항체를 1:1,000으로 희석하여 실온에서 2시간동안 배양하였으며, TBST로 3회 세척한 후 Davinch-Chemi™ Imager CAS-400SM 시스템을 이용하여 밴드를 확인하였다. 색소침착 관련 인자 단백질의 억제 활성 결과를 도 5에 나타내었다.Specifically, B16F10 cells were aliquoted in a 100 mm tissue culture dish at 1×10 6 cells/well, and then cultured for 24 hours to stabilize the cells. After removing the medium, after treatment with α-MSH (100 nm) for 2 hours, each ferment extract was replaced with a medium treated for each concentration, and after culturing for 24 hours, the medium was removed again and washed twice with 1× PBS. Each cell was dissolved in 10 ml of RIPA buffer to which 1 complete mini tab was added, and centrifuged at 4° C. and 13,200 rpm for 20 minutes. The supernatant obtained by centrifugation was quantified with a BCA protein analysis kit, and 20 μl of the protein was separated by electrophoresis on a 10% acrylamide gel. The separated protein was transferred to a polyvinylidene fluoride (PVDF) membrane using a transfer device, and then incubated in blocking buffer (5% skim milk in TBST) at room temperature for 1 hour. The primary antibody was diluted and incubated at 4° C. overnight, and then washed three times with TBST (tris-buffered saline and tween 20) at 10-minute intervals again. The secondary antibody was diluted 1:1,000 and incubated for 2 hours at room temperature, washed 3 times with TBST, and then the band was confirmed using the Davinch-Chemi™ Imager CAS-400SM system. The results of the inhibitory activity of the pigmentation-related factor protein are shown in FIG. 5 .
도 5에 나타낸 바와 같이, α-MSH에 의해 증가된 MITF, TRP-1, TRP-2 및 티로시나아제의 단백질 발현양이 단풍잎돼지풀 및 개망초 발효 추출물의 처리에 의해 감소된 것을 확인하였다. As shown in FIG. 5 , it was confirmed that the protein expression levels of MITF, TRP-1, TRP-2, and tyrosinase increased by α-MSH were reduced by the treatment of the fermented extracts of Maple leaf ragweed and fermented ragweed.
6-2. 색소침착 관련 인자의 mRNA 발현 확인6-2. Confirmation of mRNA expression of pigmentation-related factors
B16F10 세포를 100mm 컬쳐 디쉬에 시딩한 후 단풍잎돼지풀 및 개망초 발효 추출물을 농도별로 처리하고 24시간 동안 배양하였다. 배지 상등액을 제거한 후 트리졸 라이시스 버퍼를 각 웰에 1mL씩 분주하여 세포를 용해시킨 후 클로로폼 200μL를 분주하여 20초간 위아래로 흔들어주었다. 그 후 13,200rpm 에서 20분간 원심 분리하여 상층액을 이소프로판올 0.5 mL이 들어있는 튜브에 옮겨 섞었다. 다시 13,200rpm에서 20분간 원심분리 하였고, 그 상층액을 제거 한 후 75% EtOH-DEPC수를 각 튜브에 1 mL씩 분주하여 13,200rpm에서 5분간 원심 분리한 뒤 상층액을 제거한 뒤 실온에서 건조시켰다. DEPC를 50μL씩 분주하여 녹인 후 96-웰 플레이트에 RNA 5μL와 멸균수 195μL를 첨가하여 260 및 280nm에서 각각 흡광도를 측정하여 전체 RNA 양을 측정하였다. Oligo(dT) 15 프라이머(500 μg/mL) 1μL, 추출한 RNA(2 μg)와 뉴클레이즈 프리 워터로 10μL를 맞추고 75℃에서 5분간 반응시킨 후 5X 반응 완충액, MgCl2, PCR 뉴클레오티드 믹스, RNasinⓡ 억제제, 역전사 효소, 뉴클레이즈 프리 워터를 첨가하여 25℃에서 5분, 42℃에서 60분, 70℃에서 15분간 반응시켜 cDNA를 합성하였다.After the B16F10 cells were seeded in a 100 mm culture dish, the fermented extracts of maple leaf ragweed and fermented ragweed were treated by concentration and cultured for 24 hours. After removing the medium supernatant, 1 mL of trizol lysis buffer was dispensed into each well to dissolve the cells, and 200 μL of chloroform was dispensed and shaken up and down for 20 seconds. After centrifugation at 13,200 rpm for 20 minutes, the supernatant was transferred to a tube containing 0.5 mL of isopropanol and mixed. Centrifuged again at 13,200 rpm for 20 minutes, and after removing the supernatant, 1 mL of 75% EtOH-DEPC water was dispensed into each tube, centrifuged at 13,200 rpm for 5 minutes, and the supernatant was removed and dried at room temperature. . After dissolving 50 μL of DEPC, 5 μL of RNA and 195 μL of sterile water were added to a 96-well plate, and absorbance was measured at 260 and 280 nm, respectively, to measure the total RNA amount. Oligo (dT) 15 primer (500 µg/mL) 1 µL, extracted RNA (2 µg) and nuclease-free water in 10 µL, react at 75 ° C for 5 minutes, 5X reaction buffer, MgCl 2 , PCR nucleotide mix, RNasin ⓡ Inhibitor, reverse transcriptase, and nuclease-free water were added and reacted at 25° C. for 5 minutes, 42° C. for 60 minutes, and 70° C. for 15 minutes to synthesize cDNA.
MITF, 티로시나아제에 대한 mRNA 발현을 확인하기 위하여, 실시간 중합효소 연쇄반응(real time-polymerase chain reaction, RT-PCR)을 실시하였다. PCR 튜브에 5× green Go Taq Flexi 완충액, MgCl2, PCR 뉴클레오티드 믹스(10mM), 프라이머, Go Taq DNA 중합효소, 뉴클레이즈 프리 워터, 상기 합성한 cDNA를 첨가하여 혼합한 후 PCR을 실행하였다. 이때 PCR은 94℃ 30초, 56℃ 60초, 72℃ 1분씩 35 사이클을 실시하였으며, PCR 종료 후 0.002% EtBr을 첨가한 1.5% 아가로스 겔에 PCR 산물을 넣고 100V에서 40분간 전기영동하였다. 그 후 UV 투과조영기를 이용하여 밴드를 확인하여 정량분석하였다. RT-PCR 수행에 사용한 프라이머는 표 2에 나타내었으며, 색소침착 관련 인자의 mRNA 발현을 확인한 결과는 도 6에 나타내었다.In order to confirm the mRNA expression for MITF and tyrosinase, real time-polymerase chain reaction (RT-PCR) was performed. 5× green Go Taq Flexi buffer, MgCl 2 , PCR nucleotide mix (10 mM), primer, Go Taq DNA polymerase, nuclease-free water, and the synthesized cDNA were added to a PCR tube and mixed, followed by PCR. At this time, PCR was carried out for 35 cycles of 94 ℃ 30 sec, 56 ℃ 60 sec, 72
도 6에 나타낸 바와 같이, 단풍잎돼지풀 및 개망초 발효 추출물의 농도가 증가함에 따라서 MITF 및 티로시나아제의 mRNA 발현이 유의하게 억제됨을 확인하였다. As shown in FIG. 6 , it was confirmed that the mRNA expression of MITF and tyrosinase was significantly inhibited as the concentration of the fermented extract of Maple Leaf ragweed and fermented ragweed was increased.
실시예 7. 단풍잎돼지풀 및 개망초 발효 추출물과 단풍잎돼지풀 또는 개망초 추출물의 생리활성 비교Example 7. Comparison of physiological activity of maple leaf ragweed and fermented ragweed extract and maple leaf ragweed or fermented ragweed extract
7-1. 단풍잎돼지풀 또는 개망초 추출물 제조7-1. Manufacture of extract of maple leaf ragweed or ragweed
단풍잎돼지풀을 1 내지 3cm로 잘라 혼합한 후 1,300 메쉬로 분쇄하였다. 분쇄된 단풍잎돼지풀을 127℃에서 30분 동안 멸균하였다. 멸균된 단풍잎돼지풀을 수직 환류 냉각기가 부착된 추출 플라스크에 옮기고, 시료 중량의 10배수 증류수를 용매로 가한 후, 100℃에서 60분 동안 열수 추출을 3회 실시하여 단풍잎돼지풀 추출물을 제조하였다. Maple leaf ragweed was cut into 1 to 3 cm and mixed and then pulverized to 1,300 mesh. The crushed maple leaf ragweed was sterilized at 127° C. for 30 minutes. The sterilized maple leaf ragweed was transferred to an extraction flask equipped with a vertical reflux cooler, distilled
개망초 추출물도 상기 단풍잎돼지풀 추출물과 동일한 방법으로 제조하였다.The extract of aloe vera was also prepared in the same way as the extract of Maple Leaf ragweed.
7-2. 7-2. 전자공여능electron donating ability 측정 measurement
실시예 2의 전자공여능 측정방법과 동일한 방법으로 단풍잎돼지풀 추출물, 개망초 추출물과 상기 실시예 1에서 제조한 단풍잎돼지풀 및 개망초 발효 추출물의 전자공여능을 비교하였으며, 그 결과를 도 7에 나타내었다.In the same manner as in Example 2 for measuring electron donating ability, the electron donating ability of the maple leaf ragweed extract, the algae extract, and the maple leaf ragweed and fermented fermented ragweed extracts prepared in Example 1 were compared, and the results are shown in FIG. 7 .
도 7에 나타낸 바와 같이, 실시예 1에서 제조한 단풍잎돼지풀 및 개망초 발효 추출물은 단풍잎돼지풀 추출물 또는 개망초 추출물 단독에 비하여 전자공여능이 우수함을 확인하였다. 즉, 단풍잎돼지풀 또는 개망초 단독에 비해 단풍잎돼지풀과 개망초를 혼합 및 발효한 후 추출하여 수득한 단풍잎돼지풀 및 개망초 발효 추출물은 두 물질의 시너지 효과에 의하여 생리활성 효과가 현저하게 우수함을 확인하였다. As shown in FIG. 7 , it was confirmed that the fermented maple leaf ragweed extract prepared in Example 1 had superior electron donating ability compared to the maple leaf ragweed extract or the maple leaf ragweed extract alone. That is, it was confirmed that the fermented maple leaf ragweed and fermented ragweed extract obtained by mixing and fermenting maple leaf ragweed and ragweed alone, compared to maple leaf ragweed or ragweed alone, had significantly superior physiological activity effects due to the synergistic effect of the two substances. .
7-3. 엘라스타아제 억제 활성7-3. Elastase inhibitory activity
실시예 4와 동일한 방법으로 단풍잎돼지풀 추출물, 개망초 추출물과 상기 실시예 1에서 제조한 단풍잎돼지풀 및 개망초 발효 추출물의 엘라스타아제 억제 활성을 비교하였으며, 그 결과를 도 8에 나타내었다.In the same manner as in Example 4, the elastase inhibitory activity of the maple leaf ragweed extract, the A. ragweed extract, and the maple leaf ragweed extract and the fermented Amanita fermented extract prepared in Example 1 were compared, and the results are shown in FIG. 8 .
도 8에 나타낸 바와 같이, 실시예 1에서 제조한 단풍잎돼지풀 및 개망초 발효 추출물은 단풍잎돼지풀 추출물 또는 개망초 추출물 단독에 비하여 엘라스타아제 억제 활성, 즉, 주름 개선 효과가 우수함을 확인하였다. As shown in FIG. 8 , it was confirmed that the fermented maple leaf ragweed extract prepared in Example 1 was superior to the elastase inhibitory activity, that is, the wrinkle improvement effect, compared to the maple leaf ragweed extract or the ragweed extract alone.
종합적으로 이상의 실험 결과를 통하여 본 발명자들은 단풍잎돼지풀 및 개망초 발효 추출물이 티로시나아제의 활성을 억제하고, 색소 침착 관련 인자의 발현을 억제하는 효과가 우수하며, 엘라스타아제의 활성을 저해하는 등 미백 및 주름 개선 효과가 우수함을 확인하였다. 특히, 본 발명에 따른 단풍잎돼지풀 및 개망초 발효 추출물은 단풍잎돼지풀 또는 개망초 추출물 단독에 비하여 현저하게 효과가 우수한바, 피부 미백 또는 주름 개선을 위한 화장료, 피부 외용제 및 식품 분야 등에서 다양하게 활용될 수 있다.Overall, through the above experimental results, the present inventors found that the fermented extracts of maple leaf ragweed and ragweed inhibit the activity of tyrosinase, have an excellent effect of suppressing the expression of pigmentation related factors, inhibit the activity of elastase, etc. It was confirmed that the whitening and wrinkle improvement effects were excellent. In particular, the fermented maple leaf ragweed and ragweed extract according to the present invention is significantly more effective than the maple leaf ragweed or ragweed extract alone, so it can be used variously in cosmetics, external preparations for skin and food fields for skin whitening or wrinkle improvement. have.
이하, 제제예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 제제예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 제제예에 의해 제한되는 것으로 해석되지 않는다.Hereinafter, the present invention will be described in more detail through formulation examples. The formulation examples are only for illustrating the present invention, and the scope of the present invention is not to be construed as being limited by the formulation examples.
제제예 1. 화장료 제제의 제조Formulation Example 1. Preparation of a cosmetic formulation
1-1. 유연화장수 제조1-1. Manufacture of soft lotion
단풍잎돼지풀 및 개망초 발효 추출물 0.1중량%, 1,3-부틸렌글리콜 5.2중량%, 올레일알코올 1.5중량%, 에탄올 3.2중량%, 폴리솔베이트 20 3.2중량%, 벤조페논-9 2.0중량%, 카르복실비닐폴리머 1.0중량%, 글리세린 3.5중량%, 미량의 향, 미량의 방부제 및 잔량의 정제수를 혼합하여 통상의 방법으로 유연화장수를 제조하였다.0.1% by weight of fermented extract of maple leaf ragweed and algae fermented extract, 5.2% by weight of 1,3-butylene glycol, 1.5% by weight of oleyl alcohol, 3.2% by weight of ethanol, 3.2% by weight of
1-2. 밀크로션 제조1-2. Milk lotion manufacturing
단풍잎돼지풀 및 개망초 발효 추출물 0.1중량%, 글리세린 5.1중량%, 프로필렌글리콜 4.2중량%, 토코페릴아세테이트 3.0중량%, 유동파라핀 4.6중량%, 트리에탄올아민 1.0중량%, 스쿠알란 3.1중량%, 마카다미아너트오일 2.5중량%, 폴리솔베이트 60 1.6중량%, 솔비탄세스퀴롤레이트 1.6중량%, 프로필파라벤 0.6중량%, 카르복실비닐폴리머 1.5중량%, 미량의 향, 미량의 방부제, 잔량의 정제수를 혼합하여 통상의 방법으로 밀크로션을 제조하였다. Maple leaf ragweed and fermented fermented extract 0.1% by weight, glycerin 5.1% by weight, propylene glycol 4.2% by weight, tocopheryl acetate 3.0% by weight, liquid paraffin 4.6% by weight, triethanolamine 1.0% by weight, squalane 3.1% by weight, macadamia nut oil 2.5 Weight %,
1-3. 영양크림 제조1-3. Nutrient cream manufacturing
단풍잎돼지풀 및 개망초 발효 추출물 0.5중량%, 글리세린 4.0중량%, 바셀린 3.5중량%, 트리에탄올아민 2.1중량%, 유동파라핀 5.3중량%, 스쿠알란 3.0중량%, 밀납 2.6중량%, 토코페릴아세테이트 5.4중량%, 폴리솔베이트 60 3.2중량%, 카르복실비닐폴리머 1.0중량%, 솔비탄세스퀴올레이트 3.1중량%, 미량의 향, 미량의 방부제 및 잔량의 정제수를 혼합하여 통상의 방법으로 영양크림을 제조하였다.0.5% by weight of fermented extract of maple leaf ragweed and algae fermented extract, 4.0% by weight of glycerin, 3.5% by weight of petrolatum, 2.1% by weight of triethanolamine, 5.3% by weight of liquid paraffin, 3.0% by weight of squalane, 2.6% by weight of beeswax, 5.4% by weight of tocopheryl acetate, 3.2 wt% of
1-4. 마사지크림 제조1-4. massage cream manufacturing
단풍잎돼지풀 및 개망초 발효 추출물 0.5중량%, 글리세린 4.0중량%, 바셀린 3.5중량%, 트리에탄올아민 0.5중량%, 유동파라핀 24.0중량%, 스쿠알란 3.0중량%, 밀납 2.1중량%, 토코페릴아세테이트 0.1중량%, 폴리솔베이트 60 2.4중량%, 카르복실비닐폴리머 1.0중량%, 솔비탄세스퀴올레이트 2.3중량%, 미량의 향, 미량의 방부제 및 잔량의 정제수를 혼합하여 통상의 방법으로 마사지크림을 제조하였다.0.5% by weight of fermented extract of maple leaf ragweed and sagebrush, 4.0% by weight of glycerin, 3.5% by weight of petrolatum, 0.5% by weight of triethanolamine, 24.0% by weight of liquid paraffin, 3.0% by weight of squalane, 2.1% by weight of beeswax, 0.1% by weight of tocopheryl acetate, A massage cream was prepared in a conventional manner by mixing 2.4 wt% of
1-5. 세정용 바디클렌저 제조1-5. Manufacture of body cleanser for cleaning
단풍잎돼지풀 및 개망초 발효 추출물 1g, 음이온계면활성제 18g, 비이온계면활성제 5g, 글리세린 7g, 소듐클로라이드 3g, 천연올리브액상비누 1.5g, 향료 1g 및 물 100g을 혼합하여 통상의 방법으로 세정용 바디클렌저를 제조하였다.A body cleanser for cleansing by mixing 1 g of fermented maple leaf ragweed extract, 18 g of anionic surfactant, 5 g of nonionic surfactant, 7 g of glycerin, 3 g of sodium chloride, 1.5 g of natural olive liquid soap, 1 g of fragrance and 100 g of water. was prepared.
이상, 본 발명내용의 특정한 부분을 상세히 기술하였는바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적인 기술은 단지 바람직한 실시양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의해 정의된다고 할 것이다. Above, specific parts of the present invention have been described in detail, for those of ordinary skill in the art, it is clear that these specific descriptions are only preferred embodiments, and the scope of the present invention is not limited thereby. something to do. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.
Claims (8)
A cosmetic composition for skin whitening or wrinkle improvement comprising a fermented extract of hot spring water of a maple leaf ragweed ( Ambrosia trifida ) and annual fleabane mixture as an active ingredient.
The cosmetic composition for skin whitening or wrinkle improvement according to claim 1, wherein the fermentation is made by a Lactobacillus sp.
상기 온천수는 pH 7 내지 9인, 피부 미백 또는 주름 개선용 화장료 조성물.
According to claim 1,
The hot spring water has a pH of 7 to 9, a cosmetic composition for skin whitening or wrinkle improvement.
상기 온천수는 칼륨 1 내지 2mg/L, 나트륨 40 내지 50mg/L, 칼슘 5 내지 10mg/L, 실리콘 20 내지 30mg/L, 리튬 0.05 내지 0.2mg/L, 스트론튬 0.1 내지 0.5mg/L 및 마그네슘 0.1 내지 0.4mg/L을 포함하는 것인, 피부 미백 또는 주름 개선용 화장료 조성물.
5. The method of claim 4,
The hot spring water is potassium 1 to 2 mg / L, sodium 40 to 50 mg / L, calcium 5 to 10 mg / L, silicon 20 to 30 mg / L, lithium 0.05 to 0.2 mg / L, strontium 0.1 to 0.5 mg / L, and magnesium 0.1 to A cosmetic composition for skin whitening or wrinkle improvement, comprising 0.4 mg/L.
상기 추출물은 물, 탄소수 1 내지 4의 알코올 및 이들의 혼합용매로부터 선택된 1종 이상의 용매로 추출된 것인, 피부 미백 또는 주름 개선용 화장료 조성물.
According to claim 1,
The extract is a cosmetic composition for skin whitening or wrinkle improvement, which is extracted with one or more solvents selected from water, alcohols having 1 to 4 carbon atoms, and mixed solvents thereof.
A composition for external application for skin whitening or wrinkle improvement, comprising, as an active ingredient, a fermented extract of hot spring water of a mixture of maple leaf ragweed and ragweed.
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| KR102572172B1 (en) * | 2023-02-07 | 2023-08-31 | 주식회사 제이엘인터내셔널 | Skin care cosmetics with excellent viscosity and a manufacturing method thereof. |
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| KR100962587B1 (en) * | 2007-10-02 | 2010-06-11 | 동국대학교 산학협력단 | A method for fermentation of natural plants and herbal medicines, a fermented product prepared therefrom and a paharmaceutical composition, a cosmetic compositon and a food composition comprising the product |
| KR20120021766A (en) * | 2010-08-16 | 2012-03-09 | 전용진 | Liquid crystalline cream |
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