KR20200015423A - 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
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- KR20200015423A KR20200015423A KR1020190094289A KR20190094289A KR20200015423A KR 20200015423 A KR20200015423 A KR 20200015423A KR 1020190094289 A KR1020190094289 A KR 1020190094289A KR 20190094289 A KR20190094289 A KR 20190094289A KR 20200015423 A KR20200015423 A KR 20200015423A
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- KR
- South Korea
- Prior art keywords
- maple leaf
- extract
- skin whitening
- present
- fermented
- Prior art date
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Abstract
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 maple leaf ragweed and Gaecho fermented extract, and more particularly, to a composition for improving skin whitening or wrinkles 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 to general life. Especially, as skin protection is increasing as a human protective film, the interest in skin-related beauty and disease mitigation treatments is increasing due to a change in perception that quality of life is improved. In general, the preference for white skin is increasing due to the deepening of environmental pollution and skin damage caused by ultraviolet rays, and as the pursuit of clearer and clearer skin with transparent makeup, there is increasing interest in how to obtain skin whitening effect. . In addition, as the diversification and individualization of modern values stand out, the increase in interest 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 the safety of products. Is actively researching and developing 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 pole layer, and the stratum corneum. Among the various cell types that make up this, melanocytes are biopolymers of phenols that determine hair color and skin color. They synthesize and secrete melanin, a complex of black protein, to prevent skin damage caused by UV rays. It also performs several important functions, including the absorption of toxic drugs. However, excessive production of melanin causes skin diseases such as blemishes and freckles, and in severe cases, it has been reported to be associated with 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, the skin color is determined according to the content and distribution of melanin, and is related to the number and distribution of melanosomes generated in melanocytes and released to the outside of cells. Melanin is produced in the organelles of skin cells called melanosomes of melanocytes and migrates to the surrounding epidermal cells called keratinocytes. Melanin is produced by the oxidation process by tyrosinase, which oxidizes tyrosine with tyrosine as a substrate. Hyperpigmentation of the skin may be caused by various factors such as hormonal abnormalities, genetic disorders, ultraviolet irradiation, etc. after the inflammatory response of the skin, mainly due to melanin synthesis abnormalities and distribution abnormalities.
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 aimed at whitening has been conducted by regulating the activity and expression of tyrosinase enzymes from the point of view of aesthetic function, and to date, hydroquinone and ascorbic acid have been known. , Kojic acid, retinole, and arbutin. However, most of the materials exhibiting strong melanin biosynthesis inhibitory activity cause side effects such as denaturation or lethality of pigment cells, impairing the intrinsic 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.Ecological disturbances, on the other hand, are wildlife that can cause disturbances or cause disturbances in ecosystems. Invasive alien species can cause serious problems, such as encroaching native habitats, disrupting the balance of ecosystems and reducing species diversity as they enter native ecosystems. As a result, governments have regulated and regulated wildlife disturbances by law, established a management organization, or implemented an alien species ecosystem impact assessment system to determine whether imports are likely to adversely affect ecosystems.
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 under
단풍잎돼지풀(Ambrosia trifida)은 북아메리카가 원산지인 귀화식물로 여름철에 피는 꽃의 꽃가루가 알레르기성 비염이나 피부염을 일으킨다. 확산 속도도 빨라 농작물에 피해를 준다. Maple leaf scab ( Ambrosia trifida ) is a naturalized plant native to North America. Pollen of flowers blooming in summer causes allergic rhinitis or dermatitis. The rate of spread is also fast, damaging 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, Gaemancho ( Annual fleabane ) is the most common naturalized plant in Korea. The rate of spreading is so high that if we do not weed in time, it will cause severe damage to 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.Therefore, the present inventors have studied to utilize the maple leaf ragweed and forget-me-not, which are classified as ecosystem disturbance species, as a functional raw material, thereby completing the present invention by confirming the remarkable skin whitening and wrinkle improvement effect of the fermented extract of the maple leaf ragweed and forget-me-not. It became.
따라서 본 발명의 목적은, 단풍잎돼지풀 및 개망초 발효 추출물을 유효성분으로 포함하는 피부 미백 또는 주름 개선용 화장료, 피부외용제 또는 식품 조성물을 제공하는 것이다.Therefore, an object of the present invention is to provide a skin whitening or wrinkle improving cosmetic, skin external preparation or food composition comprising the maple leaf ragweed and Gaecho fermented extract 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 improving skin whitening or wrinkles, including maple leaf ragweed and Gaechocho fermented extract as an active ingredient.
또한, 본 발명은 단풍잎돼지풀 및 개망초 발효 추출물을 유효성분으로 포함하는 피부 미백 또는 주름 개선용 피부 외용제 조성물을 제공한다. The present invention also provides a skin external preparation composition for improving skin whitening or wrinkles, including a maple leaf ragweed and Gaecho fermented extract as an active ingredient.
또한, 단풍잎돼지풀 및 개망초 발효 추출물을 유효성분으로 포함하는 피부 미백 또는 주름 개선용 식품 조성물을 제공한다.In addition, the present invention provides a food composition for improving skin whitening or wrinkles, including maple leaf ragweed and Gazelweed fermented extract as active ingredients.
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.Maple leaf ragweed and Ganoderma lucidum extract according to the present invention not only has the effect of inhibiting the activity of tyrosinase and inhibiting melanogenesis in melanocytes, but also has excellent inhibitory activity of elastase, skin whitening or It may be variously used in cosmetics, skin external preparations and food fields for wrinkle improvement.
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 the fermented extract of Maple Leaf Porcine and Gaechocho according to the present invention.
Figure 2 is a view showing the results of measuring the tyrosinase inhibitory activity of the maple leaf ragweed and Gaechocho fermented extract according to the present invention.
Figure 3 is a view showing the results of measuring the elastase inhibitory activity of the maple leaf ragweed and Gaechocho fermented extract according to the present invention.
Figure 4 is a diagram showing the results of measuring the cytotoxicity of the maple leaf botanical herb and Gaechocho fermentation extract according to the present invention through the MTT analysis.
Figure 5 is a view showing the results confirming the effect on the inhibitory activity of the pigments related to the pigmentation of maple leaf botanical herb and Gaechocho fermentation according to the present invention through Western blotting.
Figure 6 is a view showing the results confirming the effect on the mRNA expression of pigmentation related factors of maple leaf ragweed and Gaechocho fermentation according to the present invention through RT-PCR.
Figure 7 is a view showing the results of comparing the electron donating ability of the maple leaf ragweed extract, Gaechocho extract and maple leaf ragweed and Gaecho fermentation extract.
Figure 8 is a view showing the results of comparing the elastase inhibitory activity of the maple leaf ragweed extract, Gaechocho extract and maple leaf ragweed and Gaecho fermented 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 an aspect of the present invention, the present invention provides a cosmetic composition for improving skin whitening or wrinkles, including as an active ingredient Maple leaf ragweed ( Ambrosia trifida ) and Gazelweed ( Annual fleabane ) fermentation.
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, "extract" is extracted from the fermentation of Maple leaf ragweed and Gazelweed using an appropriate solvent, for example, crude extract, polar solvent soluble extract or nonpolar solvent soluble extract of Maple leaf ragweed and Gazelweed It includes everything. In addition, the extract is a concept that includes all the extracts, fractions, their 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.Maple leaf ragweed and Ganoderma lucidum fermented extract which are the active ingredients of the composition of the present invention 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 maple leaf ragweed and forget-me-not fermented extract may be extracted or fractionated from various parts of the raw material plant, preferably from the leaves, stems and roots of the maple leaf ragweed and forget-me-not.
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 maple leaf ragweed and Gaecho fermented extract may be obtained by extracting, separating and fractionating from nature using extraction, separation and fractionation methods known in the art.
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 maple leaf ragweed and Gaecho fermentation extract was prepared by mixing the maple leaf ragweed and Gaechocho, followed by fermentation by adding hot spring water.
상기 단풍잎돼지풀 및 개망초는 바람직하게는 동량, 즉 1:1의 중량비율로 혼합할 수 있다. The maple leaf ragweed and forget-me-not can be preferably mixed in the same amount, that is, 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 and warm water is preferably pH 7 to 9, preferably pH 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 one 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 substances are decomposed and changed by microbial action to produce useful substances. What can be used for the fermentation in the present invention is not limited if the microorganisms that can enhance the activity of the maple leaf and the forget-me-notweed through the fermentation, preferably the strain of the genus Lactobacillus. 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 Lactobacillus bulgaricus , Lactobacillus lactis , Lactobacillus fermentum , and the like, but are not limited thereto. In an embodiment of the present invention, the microorganisms for fermentation, but mixed with Lactobacillus Lactobacillus rhamnosus ( Lactobacillus rhamnosus ) and Lactobacillus paracasei ( 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.In the present invention, the fermentation may be performed for 1 day to 5 days at 25 to 40 ℃, preferably at 2 to 4 days at 32 to 40 ℃, but is not limited thereto. In one embodiment of the present invention, after inoculating the strain of the genus Lactobacillus, by fermentation at 37 ℃ for 3 days to prepare a maple leaf ragweed and Gaechocho fermentation 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 maple leaf ragweed and Gaecho fermented extract, and any pharmaceutically acceptable organic solvent may be used. For example, as the solvent, alcohols having 1 to 4 carbon atoms such as water, methanol, ethanol, propanol, isopropanol, butanol, or the like, alone or in combination of two or more. It can be mixed and used, and water is most preferable.
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.It is preferable that the said extraction temperature is 50-100 degreeC. In addition, as the extraction method, various methods such as hot water extraction, cold extraction, reflux cooling extraction, solvent extraction, steam distillation, ultrasonic extraction, elution, and compression may be used, but are not limited thereto, and preferably, hot water extraction or The 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 maple leaf ragweed and Gaecho fermented extract may be obtained by cooling, heating and filtration at room temperature in a conventional manner known in the art, and may further evaporate, spray-dried, or lyophilized solvent. In addition, the maple leaf ragweed and Gaecho fermented extract 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 may be further purified by various chromatography such as silica gel column chromatography, thin layer chromatography, high performance liquid chromatography, and the like. You can get it.
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 maple leaf ragweed and Ganoderma lucidum fermentation extract used in the present invention is a concept including all extracts, fractions and purified products obtained at each step of extraction, fractionation or purification, 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 one embodiment of the present invention, the maple leaf ragweed and Ganoderma lucidum fermentation extracts inhibit tyrosinase activity in tyrosinase enzyme experiments, and have an excellent effect of inhibiting melanin production in a cell model induced melanin accumulation. Therefore, the maple leaf ragweed and Ganoderma lucidum fermentation extract 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, maple leaf ragweed and Gaechocho fermented extract have an excellent effect of inhibiting the activity of elastase. Therefore, the maple leaf ragweed and Gaechocho fermented extract of the present invention can be usefully used as a cosmetic composition for improving wrinkles.
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 maple leaf ragweed and Gaecho fermented extract may be included in an amount of 0.01 to 95% by weight, and more preferably 1 to 80% by weight, based on the total weight of the cosmetic composition. When the content is less than 0.01% by weight, the skin whitening or wrinkle improvement effect may be insignificant, and when the content exceeds 95% by weight, the effect increase rate compared to the amount used 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 auxiliaries and carriers such as antioxidants, stabilizers, solubilizers, vitamins, pigments, fragrances and the like that are commonly used in cosmetic compositions in addition to the active ingredient. For example, the cosmetic composition may further include auxiliary ingredients such as glycerin, butylene glycol, polyoxyethylene hardened 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 cosmetic compositions 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, spray, and the like. It is not limited to this. More specifically, it may be prepared in the form of a flexible lotion, nutrition lotion, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, mask pack, spray or 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, the carrier component may include animal oil, vegetable oil, wax, paraffin, starch, trakant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide and the like. 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, carrier components may include lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, and the like, and in particular, in the case of spray, additionally chlorofluorohydrocarbon, propane Propellants, such as butane, dimethyl ether, and the like.
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 carrier component may include a solvent, a solubilizer, an emulsion, and the like, and specifically, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, and propylene. Glycols, 1,3-butylglycol oils, glycerol aliphatic esters, polyethylene glycols, fatty acid esters of sorbitan, and the like.
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, liquid carrier diluents such as water, ethanol and propylene glycol as carrier components; Suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester; Microcrystalline cellulose, aluminum metahydroxy, bentonite, agar, tracant and the like.
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, the carrier component is an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, isethionate, an imidazolinium derivative, a methyltaurate, a sarcosinate, a fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, ethoxylated glycerol fatty acid esters, and the like.
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 skin external preparation composition for improving skin whitening or wrinkles, including maple leaf ragweed and Gaecho fermented extract as an active ingredient.
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 topical skin composition, it is further added to fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants. Common to water, ionic or nonionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or external preparations for the skin It may contain adjuvants commonly used in the field of dermatology, such as any other ingredients used as. The ingredients may also be introduced in amounts 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 in a topical skin formulation, it may have a formulation such as, but not limited to, an ointment, patch, gel, cream or spray.
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 maple leaf ragweed and Gaecho fermented extract 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 lightening 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 kind of the food. Examples of the food to which the substance can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, dairy products including gum, ice cream, various soups, beverages, tea, drink, Alcoholic beverages and vitamin complexes, and the like and include all of the health foods in the conventional 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 used as it is, or may be used together with other food or food ingredients, or may be used as appropriate according to conventional methods. The mixed amount of the active ingredient may be appropriately determined depending on 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 beverage composition. The health beverage composition may include various flavoring agents or natural carbohydrates as additional components, such as ordinary drinks, in addition to maple leaf ragweed and Gaecho fermented extract. The natural carbohydrates described above may be used as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and natural sweeteners such as dextrin and cyclodextrin, and synthetic sweeteners such as saccharin and aspartame. The proportion of said natural carbohydrates 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 salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, And carbonizing agents used in the carbonated beverage. In addition, the composition of the present invention may include a pulp for the production of natural fruit juice, fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not critical but is usually 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 with reference to Examples. These examples are only for illustrating the present invention, 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 및 개망초 발효 추출물 제조 And Ganoderma lucidum fermentation extract preparation
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 the maple leaf ragweed and forget-me-not fermented extract, cut the maple leaf ragweed and forget-me-not in 1 to 3cm, respectively, were mixed in equal amounts and then ground to 1,300 mesh. The ground maple leaf and forage mixture were sterilized at 127 ° C. for 30 minutes. To ferment the mixture, hot spring water was mixed at 1: 1 (w / v) and homogenized to prepare a hot water mixture. The components of the warm 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 to the total weight of the hot water mixture. Next, inoculated with a high concentration of lactic acid bacteria Lactobacillus strain in 10% concentration prepared by the above process at a concentration of 10%, it was fermented for 72 hours at 37 ℃, 120rpm conditions. Lactobacillus genus strain was used commercially available, 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 end of fermentation, the maple leaf and the forget-me-not fermented product were transferred to an extraction flask equipped with a vertical reflux condenser, and 10 times distilled water of the sample weight was added as a solvent, followed by extraction repeated three times at 100 ° C. for 60 minutes. To prepare a maple leaf ragweed and Gaecho fork fermented extract.
실시예 2. 단풍잎돼지풀 및 개망초 발효 추출물의 전자공여능 측정Example 2 Determination of Electron Donating Ability of Fermented Extracts from Maple Leaf Porcine Grass
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), add 60 μl of DPPH (1,1-diphenyl-2-picrylhydrazyl) and 120 μl of Maple Leaf Porcine and Ganoderma Lucidum per each concentration, mix, and leave for 15 minutes before 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 addition 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 Figure 1, the effect of increasing the electron donating ability as the concentration of the maple leaf ragweed and Gaecho fermented extract increased.
실시예Example 3. 3. 단풍잎돼지풀Maple Leaf 및 개망초 발효 추출물의 티로시나아제 저해 활성 측정 Determination of Tyrosinase Inhibitory Activity of Fermented Extracts
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 measurement to confirm the whitening effect was measured as follows. After preparing a mixed solution of 40 μl of a substrate solution of 10 mM L-DOPA (Sigma, USA) in 80 μl of 67 mM phosphate buffer solution (pH 6.8) and 40 μl of a sample solution (foliar leaf and Ganoderma lucidum extract), 200 U / ml mushroom tyrosinase was prepared. 40 μl of (Sigma, USA) was added and reacted at 37 ° C. for 10 minutes. Thereafter, absorbance was measured at 492 nm to confirm DOPA chromium formed in the reaction solution. Tyrosinase inhibitory activity was calculated as in
[수학식 2][Equation 2]
저해율(%) =(1 - 시료첨가군의 흡광도/무첨가군의 흡광도)×100% Inhibition = (1-absorbance of sample addition group / absorbance of no addition group) x 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 tyrosinase inhibitory activity was increased as the concentrations of the maple leaf herb and Ganoderma lucidum fermentation extract were increased, and thus the whitening effect was excellent.
실시예 4. 단풍잎돼지풀 및 개망초 발효 추출물의 엘라스타아제 저해 활성 측정Example 4 Determination of Elastase Inhibitory Activity of Fermented Extracts from Maple Leaf Porcine and Ganoderma Lucidum
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 anti-wrinkle effect, it was confirmed that the elitase inhibitor activity, a representative enzyme promoting skin aging. Specifically, a sample solution (foliar leaf and Ganoderma lucidum fermentation extract) was prepared to a certain concentration, and 40 μl of each was dispensed in a 96-well plate, and 2.5 U / ml pork pancreas dissolved in 50 mM tris-HCl buffer (pH8.6). After adding 40 μl of an 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) was added as a substrate. It was. The reaction was carried out at 37 ° C. for 30 minutes, and the 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, the results are shown in FIG.
[수학식 3][Equation 3]
저해율(%) =(1 - 시료첨가군의 흡광도/무첨가군의 흡광도)×100% Inhibition = (1-absorbance of sample addition group / absorbance of no addition group) x 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 Figure 3, as the concentration of the maple leaf ragweed and Gaecho fermentation extract increased, the lyasease inhibitory activity was increased, and through this it was confirmed that the wrinkle improvement effect of the fermentation extract is excellent.
실시예Example 5. 5. 단풍잎돼지풀Maple Leaf 및 개망초 발효 추출물의 세포 독성 측정 Cytotoxicity of Korean Fermented Extracts
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). B16F10 cell culture was performed using DMEM medium added with 10% FBS and 1% penicillin / streptomycin (100 U / ml), and was passaged to 37 ℃, 5% CO 2 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. In order to measure cell viability, B16F10 cells were divided into 180 μl to 1 × 10 5 cells / well in a 96-well plate, and the samples (foliar leaf and Forget-me-not fermented extract) were prepared by concentration and added 20 μl, respectively, at 37 ° C. Incubated for 24 hours in a 5% CO 2 incubator. 40 μl of the MTT solution prepared at a concentration of 2.5 mg / ml was added thereto 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 the absorbance was measured at 540 nm using an ELISA reader. Cell viability was measured as in Equation 4 below, and the results are shown in FIG. 4.
[수학식 4][Equation 4]
세포생존율(%) =(1 - 시료첨가군의 흡광도/무첨가군의 흡광도)×100Cell survival rate (%) = (1-absorbance of sample addition group / absorbance of no addition 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 Figure 4, the maple leaf botanical herb and Forget-me-not fermented extract showed no cytotoxicity such as a cell viability of about 97% up to a high concentration of 500μg / ml.
실시예 6. 단풍잎돼지풀 및 개망초 발효 추출물의 색소침착 관련 인자 억제 활성 확인Example 6 Confirmation of Inhibitory Activity of Pigmentation Related Factors of Maple Leaf Porcine and Fermented Forage Extract
6-1. 색소침착 관련 인자 단백질의 억제 활성 확인6-1. Confirmation of Inhibitory Activity of Pigmentation Related Factor Proteins
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. In order to investigate the activity of pigmentation and whitening factors such as MITF, TRP-1, TRP-2, tyrosinase, etc. at the protein level, Maple leaf and Forsythia fermented extracts of B16F10 cells at 25, 50, 100 μg / ml Treatment was done by concentration. The control group was treated with α-MSH and the overexpression of melanin, and the normal group was set with no treatment with α-MSH. At this time, the expression level of β-actin, which is a housekeeping gene having almost no difference in expression level even under various conditions of cells, was identified and quantified.
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 dispensed into 1 × 10 6 cells / well in a 100 mm tissue culture dish and cultured for 24 hours to stabilize the cells. After removing the medium and treated with α-MSH (100nm) for 2 hours, each fermentation extract was replaced with a medium treated by concentration, incubated for 24 hours, and then the medium was removed again and washed twice with 1 × PBS. Each cell was dissolved in 10 ml of RIPA buffer to which a complete mini-tap was added and centrifuged at 4 ° C. and 13,200 rpm for 20 minutes. Supernatants obtained by centrifugation were quantified with a BCA protein analysis kit, and 20 μl of protein was separated by electrophoresis on a 10% acrylamide gel. The isolated protein was transferred to a polyvinylidene fluoride (PVDF) membrane using a transfer instrument, and then incubated in blocking buffer (5% skim milk in TBST) for 1 hour at room temperature. The primary antibody was diluted and incubated overnight at 4 ° C., and then washed three times with tris-buffered saline and tween 20 (TBST) every 10 minutes. The secondary antibody was diluted 1: 1,000 and incubated for 2 hours at room temperature. After washing three times with TBST, bands were identified using a Davinch-Chemi ™ Imager CAS-400SM system. Results of inhibitory activity of pigmentation related factor proteins 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 Figure 5, it was confirmed that the amount of protein expression of MITF, TRP-1, TRP-2 and tyrosinase increased by α-MSH was reduced by the treatment of maple leaf and forage fermented extract.
6-2. 색소침착 관련 인자의 mRNA 발현 확인6-2. 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.B16F10 cells were seeded in a 100 mm culture dish, and then treated with concentrations of maple leaf and forage fermented extract, and incubated 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 13,200rpm for 20 minutes, the supernatant was transferred to a tube containing 0.5 mL of isopropanol. After centrifugation at 13,200rpm for 20 minutes, the supernatant was removed, and 1 mL of 75% EtOH-DEPC water was dispensed into each tube and centrifuged at 13,200rpm for 5 minutes, and then the supernatant was removed and dried at room temperature. . 50 μL of DEPC was dispensed and dissolved, and then 5 μL of RNA and 195 μL of sterile water were added to a 96-well plate, and the absorbance was measured at 260 and 280 nm, respectively. 1 μL of Oligo (dT) 15 primer (500 μg / mL), 10 μL of extracted RNA (2 μg) and nuclease free water, and reacted at 75 ° C. for 5 minutes, followed by 5X reaction buffer, MgCl 2 , PCR nucleotide mix, RNasin The cDNA was synthesized by adding the inhibitor, reverse transcriptase and nucleated free water at 5 ° C. for 5 minutes, at 42 ° C. for 60 minutes, and at 70 ° C. for 15 minutes.
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 mRNA expression for MITF and tyrosinase, a real time polymerase chain reaction (RT-PCR) was performed. PCR was performed by adding 5 × green Go Taq Flexi buffer, MgCl 2 , PCR nucleotide mix (10 mM), primers, Go Taq DNA polymerase, nucleated free water, and cDNA synthesized above to the PCR tube. At this time, PCR was performed for 35 cycles of 94 ° C. 30 seconds, 56 ° C. 60 seconds, and 72 ° C. for 1 minute. After completion of PCR, PCR products were added to 1.5% agarose gel to which 0.002% EtBr was added and electrophoresed at 100V for 40 minutes. The band was then quantitatively analyzed using a UV penetrometer. Primers used to perform RT-PCR are shown in Table 2, and the results of confirming mRNA expression of pigmentation related factors are shown in FIG.
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 mRNA expression of MITF and tyrosinase was significantly inhibited as the concentrations of the maple leaf and Ga. Fermented extracts increased.
실시예 7. 단풍잎돼지풀 및 개망초 발효 추출물과 단풍잎돼지풀 또는 개망초 추출물의 생리활성 비교Example 7 Comparison of Physiological Activities of Fermented Extracts from Maple Leaf and Forget-weed
7-1. 단풍잎돼지풀 또는 개망초 추출물 제조7-1. Manufacture of Maple Leaf Pigweed or Ganoderma Lucidum Extract
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. Cut the maple leaf pork to 1-3cm, mixed and pulverized to 1,300 mesh. The pulverized maple leaf was sterilized for 30 minutes at 127 ℃. The sterilized maple leaf ragweed was transferred to an extraction flask equipped with a vertical reflux condenser, and 10 times distilled water of the sample weight was added as a solvent, followed by performing hot water extraction three times at 100 ° C. for 60 minutes to prepare a maple leaf extract.
개망초 추출물도 상기 단풍잎돼지풀 추출물과 동일한 방법으로 제조하였다.Cauliflower extract was prepared in the same manner as the maple leaf ragweed extract.
7-2. 7-2. 전자공여능Electron donating ability 측정 Measure
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 the method of measuring the electron donating ability of Example 2, the electron donating ability of the maple leaf ragweed extract, Gaechocho extract and the maple leaf ragweed and Gazelweed fermented extract prepared in Example 1 were compared, and the results are shown in FIG.
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, the maple leaf ragweed and Ganoderma lucidum fermented extract prepared in Example 1 were confirmed to have excellent electron donating ability as compared to the maple leaf ragweed extract or Gazelweed extract alone. In other words, compared to maple leaf ragweed or gazelweed alone, maple leaf ragweed and fermented green leaf extract obtained by mixing and fermenting maple leaf ragweed and Gazelweed were confirmed that the physiological activity effect was remarkably excellent by 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 was compared with the elastase inhibitory activity of the maple leaf ragweed extract, Gaechocho extract and the maple leaf ragweed and Gaecho fermented extract prepared in Example 1, the results are shown in FIG.
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, the maple leaf ragweed and Ganoderma lucidum fermentation extract prepared in Example 1 were found to have an excellent elastase inhibitory activity, that is, wrinkle improvement, as compared to the maple leaf ragweed extract or Gazelweed 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.Based on the above experimental results, the present inventors have shown that the fermented maple leaf and Forget-me-not fermented extracts inhibit the activity of tyrosinase, inhibit the expression of pigmentation-related factors, inhibit the activity of elastase, and the like. It was confirmed that the whitening and wrinkle improvement effect is excellent. In particular, the maple leaf ragweed and Gaechofer fermented extract according to the present invention is significantly superior to the maple leaf ragweed or Gazelweed extract alone, it can be used in a variety of cosmetics, skin external preparations 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 with reference to the 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 Cosmetic Formulation
1-1. 유연화장수 제조1-1. Flexible Cosmetics Manufacturing
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 Maple Leaf and Fermented Forage 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 polysorbate, 2.0% by weight of benzophenone-9, 1.0% by weight of carboxyvinyl polymer, 3.5% by weight of glycerin, trace amount of fragrance, trace amount of preservative, and residual amount of purified water were mixed to prepare flexible cosmetic water in a conventional manner.
1-2. 밀크로션 제조1-2. Wheat Crop Maker
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. 0.1% by weight of Maple Leaf and Fermented Fermented Leaf, Glycerin 5.1%, Propylene Glycol 4.2%, Tocopheryl Acetate 3.0%, Liquid Paraffin 4.6%, Triethanolamine 1.0%, Squalane 3.1%, Macadamia Nut Oil 2.5 Weight%,
1-3. 영양크림 제조1-3. Nutrition Cream
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 Maple Leaf and Fermented Fermented Leaf, 4.0% by weight of Glycerin, 3.5% by weight of Vaseline, 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, A nutrition cream was prepared in a conventional manner by mixing 3.2% by weight of
1-4. 마사지크림 제조1-4. Massage cream manufacturer
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 Maple Leaf and Fermented Forage Extract, 4.0% by weight of glycerin, 3.5% by weight of petroleum jelly, 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, Massage cream was prepared in a conventional manner by mixing 2.4% by weight 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.Fermented extract of Maple Leaf and Ganoderma lucidum 1g, Anionic surfactant 18g, Nonionic surfactant 5g, Glycerin 7g, Sodium chloride 3g, Natural olive liquid soap 1.5g, Flavor 1g and Water 100g 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. As mentioned above, specific parts of the present invention have been described in detail, and it is apparent to those skilled in the art that such specific descriptions are merely preferred embodiments, and thus the scope of the present invention is not limited thereto. something to do. Therefore, the substantial scope of the present invention will be defined by the appended claims and their equivalents.
Claims (8)
A cosmetic composition for improving skin whitening or wrinkles comprising maple leaf ( Ambrosia trifida ) and Gaechocho ( Annual fleabane ) fermented extract as active ingredients.
According to claim 1, The fermentation is made of a strain of the genus Lactobacillus, cosmetic composition for skin whitening or wrinkle improvement.
상기 발효는 단풍잎돼지풀 및 개망초 혼합물에 온양온천수를 더 첨가한 후 이루어진 것인, 피부 미백 또는 주름 개선용 화장료 조성물.
The method of claim 2,
The fermentation will be made after adding hot spring water to the maple leaf ragweed and Gaechocho mixture, skin cosmetic or wrinkle improvement cosmetic composition.
상기 온양온천수는 pH 7 내지 9인, 피부 미백 또는 주름 개선용 화장료 조성물.
The method of claim 3,
The hot spring water is pH 7 to 9, cosmetic composition for improving skin whitening or wrinkles.
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 0.2mg/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.
The method of claim 4, wherein
The hot spring water is potassium 1-2 mg / L, sodium 40-50 mg / L, calcium 5-10 mg / L, silicon 20-30 mg / L, lithium 0.05-0.2 mg / L, strontium 0.1-0.5 mg / L and magnesium 0.1 Containing to 0.4 mg / L, cosmetic composition for improving skin whitening or wrinkles.
상기 추출물은 물, 탄소수 1 내지 4의 알코올 및 이들의 혼합용매로부터 선택된 1종 이상의 용매로 추출된 것인, 피부 미백 또는 주름 개선용 화장료 조성물.
The method of claim 1,
The extract is extracted with water, one or more solvents selected from alcohols having 1 to 4 carbon atoms and mixed solvents thereof, cosmetic composition for skin whitening or wrinkle improvement.
Skin whitening or anti-wrinkle composition for improving skin whitening or wrinkles including fermented maple leaf and fermented forage.
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KR20220160325A (en) * | 2021-05-27 | 2022-12-06 | (주)아로마뉴텍 | Ambrosia trifida supercritical extract and its use |
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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KR20080023827A (en) * | 2006-09-12 | 2008-03-17 | 바이오스펙트럼 주식회사 | A skin whitening compositions comprising natural plant extract |
KR20090034241A (en) * | 2007-10-02 | 2009-04-07 | 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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KR20080023827A (en) * | 2006-09-12 | 2008-03-17 | 바이오스펙트럼 주식회사 | A skin whitening compositions comprising natural plant extract |
KR20090034241A (en) * | 2007-10-02 | 2009-04-07 | 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 |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
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KR20220160325A (en) * | 2021-05-27 | 2022-12-06 | (주)아로마뉴텍 | Ambrosia trifida supercritical extract and its use |
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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