KR20140132624A - Cosmetic composition for skin whitener containing fermented extract of green plum and method thereof - Google Patents
Cosmetic composition for skin whitener containing fermented extract of green plum and method thereof Download PDFInfo
- Publication number
- KR20140132624A KR20140132624A KR1020130052093A KR20130052093A KR20140132624A KR 20140132624 A KR20140132624 A KR 20140132624A KR 1020130052093 A KR1020130052093 A KR 1020130052093A KR 20130052093 A KR20130052093 A KR 20130052093A KR 20140132624 A KR20140132624 A KR 20140132624A
- Authority
- KR
- South Korea
- Prior art keywords
- extract
- skin
- cosmetic composition
- fermented extract
- green plum
- Prior art date
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Classifications
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
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Abstract
Description
본 발명은 청매실 발효 추출물을 함유하는 피부미백용 화장료 조성물 및 그 제조방법에 관한 것이다.The present invention relates to a cosmetic composition for skin whitening containing a browning room fermented extract and a method for producing the same.
특히, 본 발명은 청매실 추출물의 각종 식물성 유기산 성분은 피부의 죽은 세포로 구성된 각질을 필링하는 효능이 AHA, BHA 보다 매우 안전하고 효과적이며 발효를 통해 저분자화된 청매실 미백 유효성분들이 빠르게 피부속으로 침투할 수 있는 피부미백용 화장료 조성물 및 그 제조방법에 관한 것이다.
In particular, the present invention is based on the fact that various phytogenic organic acid components of Cheongpilmil extract are more safe and effective than peptides of AHA and BHA in peeling keratin composed of dead cells of skin, To a cosmetic composition for skin whitening and a method for producing the same.
피부는 외부의 자극으로부터 우리의 몸을 보호하고 있는 인체를 구성하는 기관들 중 가장 큰 기관으로서, 내부의 수분 및 유용성 성분의 피부 밖으로의 유출을 막아주는 장벽기능, 온도 조절기능, 배설기능, 감각기능, 호흡기능 등 다양한 생리적 역할을 담당하고 있다.Skin is the largest organ of the human body that protects our body from external stimuli. It has barrier function to prevent the internal moisture and oil components from flowing out of the skin, temperature control function, excretion function, sensation Function, and respiration function.
이러한 피부는 표피, 진피 및 피하지방층으로 구성되어 있으며 피부의 최외곽층인 표피는 외부환경으로부터 피부를 보호하는 기능을 수행한다.
The skin is composed of the epidermis, the dermis and the subcutaneous fat layer, and the epidermis which is the outermost layer of the skin functions to protect the skin from the external environment.
한편, 표피 가장 바깥층의 각질화된 죽은 세포가 세포간 지질 등의 시멘트(Cement) 기능의 도움을 받아 피부보호막으로서 기능을 하는 한편, 비정상적으로 과다하게 축적되어 거칠고 둔탁한 피부결을 만들고, 탄력이 없이 뻣뻣하며 피부색이 윤기있지 못하고 퇴색한 듯 보이는 등 미용적인 문제 뿐 아니라, 피부 노화와 주름살의 생성, 피부 건조화 현상으로 피부 건강을 저해하는 현상이 발생하기도 한다.
On the other hand, the keratinized dead cells on the outermost layer of the epidermis function as a skin barrier with the help of the cement function such as intercellular lipids, and accumulate abnormally excessively to make rough and dull skin texture, In addition to cosmetic problems such as skin color not appearing to be glazed and fading, skin aging, wrinkle formation, and dryness of the skin may cause skin health impairment.
표피층은 크게 기저층, 유극층, 과립층, 투명층, 각질층으로 나뉘어지며 각질세포는 기저층으로부터 만들어져, 피부 최외각층인 각질층에서 소실되기까지 28일을 주기로 하여 사라지고 다시 만들어지는 과정을 거치면서 피부의 건강을 유지하게 된다.The epidermis is divided into the basal layer, the superficial layer, the granular layer, the transparent layer, and the stratum corneum. The keratinocytes are made from the basal layer and disappear from the stratum corneum of the outermost layer, disappearing every 28 days, .
또한, 표피층을 구성하는 세포로는 케라티노사이트, 랑겔한스 세포, 머켈 세포, 멜라노사이드 등의 다양한 세포들이 존재하며 이중에서 대부분을 차지하는 세포가 케라티노사이트이다. 케라티노사이트는 기저층에서 세포 증식활동을 하여 많은 피부세포를 만들어 밀어올리며, 이는 유극층을 지나면서 세포의 생명을 잃으면서 내부의 세포구성성분은 세포밖으로 내보내진체 비교적 단단한 케라틴만이 최종적으로 단단하게 남은 채 최종적으로 피부 각질층에 도달한다. 그러나 이러한 케라티노사이트의 활성에 문제가 발생하면 피부 노화와 주름살의 생성, 피부 건조화 현상이 일어나 피부 늘어짐 현상을 초래해 피부의 탄력이 떨어지게 된다. 따라서 피부를 건강하고 윤택하고 아름답게 유지하기 위해서는 피부의 표피층을 구성하는 성분을 만들어내는 케라티노사이트를 활성화 시켜주고 각질층의 기능을 향상시켜줄 필요가 있다.
In addition, as the cells constituting the epidermal layer, there are various cells such as keratinocytes, Rangelans cells, Merkel cells and melanocytes, and keratinocytes are the cells that occupy most of them. The keratinocytes undergo cell proliferation in the basal layer to produce a large number of skin cells, which, in turn, lose their cell life as they pass through the anolyte layer, where the cellular constituents of the inner part are exported out of the cell. Finally, the skin reaches the stratum corneum. However, if there is a problem in the activity of keratinocytes, skin aging, wrinkles and dryness of the skin may occur, resulting in skin sagging, and the elasticity of the skin is lowered. Therefore, it is necessary to activate the keratinocyte which produces the constituent of the epidermal layer of the skin and to improve the function of the stratum corneum to maintain the skin healthy, moisturized and beautiful.
각질 세포의 각화 과정을 촉진시켜 주는 대표적인 물질로는 알파하이드록시 액시드(AHA)가 있다. AHA는 각화 촉진제로써 많이 이용되는 물질이지만 피부에 많은 량이 도포되면 피부자극을 유발할 뿐만 아니라 피부의 건조와 투명층 생성을 유발하기도 하며 이를 포함하는 화장료 조성물의 제형에서도 제형의 안정화에 좋지 않은 영향을 주는 문제점을 가지고 있다.
Alpha hydroxy acid (AHA) is a typical substance that promotes keratinocyte keratinization. AHA is a substance widely used as a keratinization promoter, but when a large amount is applied to the skin, it causes not only skin irritation, but also drying of the skin and generation of a transparent layer. Also, formulation of a cosmetic composition containing the AHA has a problem Lt; / RTI >
천연물로부터 피부 미백제에 개발을 위하여 많은 연구가 이루어지고 있으며, 그 중 일부는 제품으로 상용화 되어있다. 미백제는 멜라노사이트(melanocyte)에서 생성, 배출된 멜라닌의 흑화 과정을 막거나 티로시나제(tyrosinase)의 활성을 억제하여 멜라닌의 합성을 억제하는 효과를 갖거나 또는 생성된 흑색 멜라닌의 환원을 촉진하여 최초의 엷은 색의 멜라닌으로 되돌아가도록 한다.
Many studies have been conducted to develop skin whitening agents from natural products, and some of them have been commercialized as products. The whitening agent has the effect of inhibiting melanin synthesis by inhibiting the blackening process of melanin produced or released from melanocytes or inhibiting the activity of tyrosinase or promoting the reduction of melanin, Return to light melanin.
멜라닌은 피부, 머리카락 및 눈동자의 색에 관여하는 색소 중의 하나이다. 멜라닌은 자연의 동식물계에 널리 분포되어 있는 유황화합물로서 시스테인과 글루타치온 및 철(Fe)의 함유에 따라 갈색에서 흑색의 다양한색의 농도를 갖는 중화합체이다. 일반적으로 알카리에 난용성인 흑색 멜라닌과 가용성 적색 및 황색 멜라닌으로 구분된다.
Melanin is one of the pigments involved in the color of skin, hair and eyes. Melanin is a sulfur compound widely distributed in natural animal and plant systems. It is a neutralized substance having various colors of brown to black depending on the contents of cysteine, glutathione and iron (Fe). It is generally distinguished from alkaline-soluble black melanin and soluble red and yellow melanin.
매실나무라고도 부르는 매화나무는 장미과의 낙엽활엽 소교목으로 원산지는 중국이며, 정원수나 과수로 심는다. 높이는 4-5m이고, 잎은 어긋나며 달걀형으로 가장자리에 톱니가 있다. 향기가 좋은 흰색 또는 연분홍색의 꽃이 이른 봄에 잎보다 먼저 핀다. 열매는 매실이라고 하는데, 6-7월에 황색으로 익으며 신맛이 강하다. 매실은 주성분인 구연산의 함량이 높아 피부 각질을 안전하게 박리하는 효과이다.The plum tree, also called plum tree, is a deciduous broad-leaved arboreous tree of Rosaceae, originating in China and planted with planted trees or fruit trees. The height is 4-5m, the leaves are alternate, egg-shaped, with sawtooth on the edge. A fragrant white or pale pink flower blooms earlier than the leaves in early spring. The fruit is called plum, and it ripens in yellow color in June-July and it is strong in sour taste. The plum has high content of citric acid as its main ingredient, so it can safely exfoliate the skin.
매실에는 주성분으로서 구연산, 사과산, 호박산, 주석산 등의 유기산과 각종 미네랄이 풍부하게 함유되고 있다. 특히 매실 추출물은 60%가 구연산을 중심으로 한 유기산이다. 이러한 구연산은 -하이드록시산의 일종으로 분자내의 카르복시 그룹에 대한 -위치의 탄소원에 알콜성 하이드록시 그룹을 갖는 화합물로서 이러한 화합물들은 건성 피부, 어린선, 소포성 과각층의 제거에 유효한 성분으로 알려져 있다.Plum is rich in organic acids such as citric acid, malic acid, succinic acid, tartaric acid, and various minerals as main components. Especially, 60% of plum extract is organic acid centered on citric acid. Such citric acid is a kind of hydroxy acid, which is a compound having an alcoholic hydroxy group at a carbon atom at a position relative to the carboxy group in the molecule. Such compounds are known as effective ingredients for the removal of dry skin, lining, have.
그러나 아직까지 이러한 청매실의 발효추출물을 함유한 피부 미백용 화장품 조성물에 관한 효과 연구는 매우 미미하였다.
However, the effect on the skin whitening cosmetic composition containing the fermented extract of this browning room has been very limited.
상기와 같은 문제점을 해결하고자 창출된 본 발명의 목적은, 매실 추출물 발효에 의해 발효 과정 생성된 유기산의 안전하면서도 저분자화된 매실 의 유효성분들이 우수한 피부 각질 박리 효능과 피부 침투가 용이해 피부 각질 박리 효능과 피부미백가 탁월한 청매실 발효 추출물을 함유하는 피부미백용 화장료 조성물 및 그 제조방법을 제공하는 것을 목적으로 한다.
It is an object of the present invention, which is created to solve the above problems, to provide a safe and low-molecular-weight active ingredient of an organic acid fermented by the fermentation of plum extract, It is an object of the present invention to provide a cosmetic composition for whitening skin containing fermented extract of Cheongmiljil which has excellent efficacy and skin whitening effect and a method for producing the same.
이러한 본 발명의 목적은, 건조 매실 분말과, 상기 건조 매실 분말 중량의 약 1 ~ 10배 중량의 물을 용매로 20 ~ 60℃의 추출 온도에서 약 12 ~ 24시간 동안 저온 추출방법으로 2회 ~ 3회 반복하여 추출물을 수득한 후, 감압여과하고 여과한 추출물을 0.2um 필터로 제균하고 제균된 추출물에 캐피어 균주(Kazachstania unispora) 배양액을 1 ~ 10% 접종하여 20 ~ 25℃에서 4 ~ 7일간 배양하여 청매실 발효 추출물을 수득한 후, 감압여과하고 동결 건조하여 수득한 청매실 발효 추출물 동결건조 분말을 유효성분으로 함유하는 청매실 발효 추출물을 함유하는 피부미백용 화장료 조성물 제조방법에 의해 달성될 수 있다.
The object of the present invention is to provide a process for producing a dried beverage comprising the steps of: (1) drying a dried meju powder and water having a weight of about 1 to 10 times the weight of the dried bean powder at a temperature of 20 to 60 ° C for about 12 to 24 hours, The extract was filtered with a reduced pressure, filtered with a 0.2-μm filter, and inoculated with 1 ~ 10% of a culture of Kazachstania unispora to the inactivated extract. The present invention can be achieved by a method for producing a skin whitening cosmetic composition comprising a browning room fermentation extract containing a lyophilized fermented extract freeze-dried powder obtained by filtration under reduced pressure and lyophilization obtained as an effective ingredient .
본 발명의 다른 목적은, 위의 방법에 의해 제조되는 청매실 발효 추출물을 함유하는 피부미백용 화장료 조성물에 의해 달성될 수 있다.
Another object of the present invention can be attained by a skin whitening cosmetic composition containing a browning room fermentation extract prepared by the above method.
본 발명의 또 다른 목적은, 위의 방법에 의해 제조된 화장료 조성물을 0.01 ~ 10중량% 포함하는 미백 화장수, 미백 크림 및 팩으로 구성된 군에서 임의로 선택된 하나의 제형인 것을 특징으로 하는 청매실 발효 추출물을 함유하는 피부미백용 화장료 조성물에 의해 달성될 수 있다.
It is still another object of the present invention to provide a chewing gum fermentation extract, which is a formulation selected from the group consisting of a whitening lotion, a whitening cream, and a pack containing 0.01 to 10% by weight of a cosmetic composition prepared by the above method The present invention provides a cosmetic composition for skin whitening comprising
이상과 같은 본 발명은 피부 각질 박리와 피부 미백효능 화장료 조성물은 천연 성분을 사용함으로써 피부에 안전하면서도, 피부 각질 박리와 우수한 피부 미백 효능을 발휘할 수 있는 효과가 있다.
The present invention has the effect of exerting skin peel and skin whitening efficacy while being safe to the skin by using a natural ingredient.
도 1은 본 발명의 청매실 추출물 및 발효 추출물의 DPPH 소거능을 나타내는 그래프이다.
도 2는 멜라닌을 합성하는 세포주에 대한 세포 독성을 나타내는 그래프이다.
도 3은 멜라닌세포에서 청매실 추출물과 청매실 발효추출물의 멜라닌 생성억제 효과를 나타내는 그래프이다.
도 4는 청매실 발효 추출물의 멜라닌 형성 단백질 발현 효과를 나타내는 도면이다.
도 5는 청매실 발효 추출물의 B16-F10 셀에서 멜라노젠 MEK, ERK1/2, 및 Akt 단백질 발현효과를 나타내는 도면이다.FIG. 1 is a graph showing DPPH scavenging activity of the Cheuremilm extract and fermented extract of the present invention.
2 is a graph showing cytotoxicity against melanin-synthesizing cell lines.
FIG. 3 is a graph showing the melanin production inhibitory effect of the browning room extract and the browning room fermentation extract in melanocytes.
FIG. 4 is a graph showing the effect of melanin-forming protein expression in the browning chamber fermentation extract. FIG.
FIG. 5 is a graph showing the effect of Melanogen MEK, ERK1 / 2, and Akt protein expression in B16-F10 cells of a browning chamber fermentation extract. FIG.
이하, 본 발명의 실시예를 도면을 참조하여 설명하기로 한다.Hereinafter, embodiments of the present invention will be described with reference to the drawings.
청매실 추출물 속에 함유되어 있는 구연산과 여러 가지 과일산 들이 피부의 표피에 작용하여 각질 제거를 촉진하는 피부 화장료 조성물이 알려져 있으나, 본 출원의 발명자들은 하기 실시예에 입증된 바와 같이, 매실 추출물 자체 보다 매실 추출물을 효모 또는 캐피어 균주(Kazachstania unispora)를 이용한 배양 발효를 통해 청매실의 미백효능을 유효 성분으로 하는 경우, 피부가 각질화 되어 죽은 세포로 이루어진 피부 각질의 박리효과로 청매실 발효 성분 중 미백효능 성분이 피부속으로 침투가 용이해서 현저하게 피부 미백효능이 상승될 수 있음을 확인하였다.
Citric acid and various fruit acids contained in the extract of Cheongpilmil act on the epidermis of the skin to promote exfoliation. However, the inventors of the present application have found that the plums extract When the extract is used as an active ingredient for the whitening effect of the browning room through fermentation using yeast or Kazachstania unispora, the exfoliation effect of the skin keratin, which is formed by dead skin cells, results in the whitening effect ingredient It was confirmed that the penetration into the skin was easy and the skin whitening efficacy could be remarkably increased.
본 발명의 추출물을 수득하는 방법을 상세히 설명한다.The method of obtaining the extract of the present invention will be described in detail.
본 발명의 추출물은 건조 매실 분말 그 중량의 약 1 ~ 10배, 바람직하게는 약 5 ~ 10배 중량의 물을 용매로 20 ~ 60℃의 추출 온도에서 약 12 ~ 24시간 동안 저온 추출방법 2회 ~ 3회 반복하여 추출물을 수득한 후, 감압여과하고 여과한 추출물을 0.2 um 필터로 제균하고 제균된 추출물에 캐피어 균주(Kazachstania unispora) 배양액을 1 ~ 10% 접종, 20 ~ 25℃에서 4 ~ 7일간 배양하여 청매실 발효 추출물을 수득한 후, 감압여과하고 동결 건조하여 수득한 청매실 발효 추출물 동결건조 분말을 유효성분으로 함유하는 피부 미백효능 화장료 조성물에 관한 것이다.
The extract of the present invention is prepared by dissolving water having a weight of about 1 to 10 times, preferably about 5 to 10 times the weight of the dried plum powder by a solvent at an extraction temperature of 20 to 60 ° C for about 12 to 24 hours The extract was filtered with a reduced pressure, and the filtered extract was sterilized with a 0.2-μm filter. To the inactivated extract, 1 to 10% of a culture solution of a culture of Kazachstania unispora was inoculated, The present invention relates to a skin whitening cosmetic composition containing as an active ingredient an lyophilized fermented extract lyophilized powder obtained by culturing for 7 days to obtain a fermented extract of Cheongmil chamber, followed by filtration under reduced pressure and lyophilization.
상기 유효 성분은 피부 각질 제거 효과를 충분하게 달성할 수 있으며 특별히 제한되지 않으나, 예를 들어 조성물 전체 중량을 기준으로 0.01 ~ 10 중량%로 포함될 수 있으며, 바람직하게 0.01 ~ 5 중량%로 포함될 수 있다. 상기 유효 성분이 0.01 중량% 미만으로 포함되면, 각질제거의 실질적인 효과를 얻기 어려우며, 10 중량%를 초과하여 포함되면 제품 안전성에 좋지 않은 영향을 미친다.
The active ingredient can sufficiently attain a skin exfoliating effect and is not particularly limited. For example, the active ingredient may be contained in an amount of 0.01 to 10% by weight, preferably 0.01 to 5% by weight, based on the total weight of the composition . If the active ingredient is contained in an amount of less than 0.01% by weight, it is difficult to obtain a substantial effect of exfoliation, and if it exceeds 10% by weight, it adversely affects product safety.
하나의 실시예에서, 상기 유효 성분은 청매실 발효 추출물일 수 있으며, 본 출원의 발명자들은 하기 실험예를 통해 입증된 바와 같이 청매실 발효 추출물을 안전한 저 자극 피부 각질 제거 와 피부 미백용 화장료 조성물의 유효 성분으로 사용하는 경우, 피부가 각질화 되어 죽은 세포로 이루어진 각질층을 자극없이 안전하게 각질을 박리시키고, 발효를 통해 저분자화된 청매실 미백 유효성분들이 빠르게 피부속으로 침투할 수 있어 청매실 발효 추출물의 더욱 향상된 피부미백 효과를 확인하였다.
In one embodiment, the active ingredient may be a browning room fermentation extract, and the inventors of the present application have found that the extract of the browning room fermentation extract can be safely applied to safe hypoallergenic skin exfoliation and skin care whitening cosmetic composition It is possible to safely remove the horny layer from the dead skin cells without any stimulation and to allow the low molecular weight chelating agent for whitening effective ingredients to rapidly penetrate into the skin through the fermentation, The effect was confirmed.
상기 피부 미백 화장료 조성물은 통상적인 기초 화장료, 예를 들면 미백 화장수(스킨로션), 영양로션, 미백 에센스, 팩 등에 첨가되어 제형화 될 수 있다.
The skin whitening cosmetic composition may be formulated by adding to a common basic cosmetic, for example, a whitening lotion (skin lotion), a nutrition lotion, a whitening essence, a pack and the like.
이하, 하기 실시예에 의하여 본 발명을 더욱 상세하게 설명하고자 한다. 단, 하기 실시예는 본 발명을 예시하기 위한 것일 뿐 본 발명의 범위가 이들만으로 한정되는 것은 아니다.
Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are intended to illustrate the present invention, but the scope of the present invention is not limited thereto.
실험예 1)Experimental Example 1) 청매실 발효 추출물의 각질 제거 효과The exfoliation effect of fermentation extract
1명의 건강한 여성(30 ~ 40세 사이)을 대상으로 하여 워싱오일로 세척하여 도포전 피부 밝기 정도인 L값(L-value)을 측정하고, 청매실 추출물, 청매실 발효 추출물 및 콘트롤을 정하여, 각각 1일 3회씩 도포하였다. 그 후 계속해서 도포하면서 이틀 간격으로 피부 밝기 정도인 L값(L-value)을 측정하고, 이 실험을 일주일간 실시하였다. L 값이 높아지는 것은 각질이 제거되는 정도를 보여주며, 그 결과는 하기 표 1 및 도 1에 나타내었다.One healthy woman (between 30 and 40 years old) was washed with washing oil to measure the L value (L-value) of skin brightness before application, and the browning room extract, browning room fermentation extract and control were determined to be 1 3 times a day. The L value (L-value), which is the degree of skin brightness, was measured at intervals of two days while applying continuously, and this experiment was performed for one week. The increase in the L value shows the extent to which the keratin is removed, and the results are shown in Table 1 and FIG.
(실시예2) 청매실 추출물 및 발효 추출물의 유기산 분석 (Example 2) Analysis of Organic Acid of Cheongmilil Extract and Fermented Extract
청매실 추출물, 발효 추출물의 유기산 분석은 청매실 추출물, 발효추출물을 16,000rpm 30분 원심분리하여 상등액을 0.45um 필터하여 HPLC로 분석하였으며, 그 결과는 표 2와 같다.In order to analyze the organic acids in the browning room extract and the fermentation extract, the browning room extract and the fermentation extract were centrifuged at 16,000 rpm for 30 minutes, and the supernatant was subjected to 0.45 μm filtration and analyzed by HPLC.
(실시예 3) 청매실 추출물 및 발효 추출물의 항산화 할성 (Example 3) Antioxidative activity of the extract of Cheolmeol and fermented extract
전자공여작용은 Blois의 방법을 변형하여 1,1-diphenyl-2-picrylhydrazyl에 대한 전자공여 효과를 측정하였다. 에탄올에 5 mg/100mL 농도로 제조한 DPPH 용액 2 mL에 농도를 조절한 시료 추출물 1 mL를 넣어 혼합한 다음 실온에서 20분간 반응시킨 후 525 nm에서 흡광도를 측정하였다. 대조구로 사용한 Ascorbic acid 의 농도는 0.1% 되게 조제하였으며, 건조 청매실 추출 동결 분말, 청매실 발효 추출물 동결건조 분말은100, 300, 500, 700, 1000ug/ml 농도로 황산화 효과를 측정하였으며 그 결과를 도 2에 나타내었다.
The electron donating effect was determined by modifying the Blois method and measuring the electron donating effect on 1,1-diphenyl-2-picrylhydrazyl. To 2 mL of the DPPH solution prepared at a concentration of 5 mg / 100 mL in ethanol, 1 mL of the sample extract adjusted to the concentration was added, and the mixture was reacted at room temperature for 20 minutes, and the absorbance was measured at 525 nm. The concentration of ascorbic acid was 0.1%, and the sulfation effect was measured at 100, 300, 500, 700, and 1000 ug / ml concentration of the freeze - dried powder extracted from the dried browning chamber and the freeze - dried powder of the browning chamber fermented extract. Respectively.
(실시예 4) 청매실 발효 추출물의 미백효과 (Example 4) Whitening effect of fermented extract of Cheongmil room
세포주 배양Cell culture
실험에 사용한 세포는 B16-F10 세포주로 murine melanocyte이며 멜라닌을 합성하는 세포주이다. 멜라닌 생성을 저해하는 실험을 하기 위해서 먼저 소재 2가지(청매실 열수, 청매실 열수 발효추출물)의 서로 다른 농도(0.1mg/ml, 0.5mg/ml, 1mg/ml, 1.5mg/ml)로 12시간 처리 후 CCK-8 assay solution을 50ul 넣은 후 37℃ CO2 인큐베이터(incubator)에서 3시간 반응 후 OD450nm 파장에서 값을 확인하였으며, 그 결과를 도 1에 나타내었다.
The cells used for the experiment are murine melanocytes of B16-F10 cell line and are a cell line which synthesizes melanin. To test for the inhibition of melanogenesis, first treatment was carried out for 12 hours at different concentrations (0.1 mg / ml, 0.5 mg / ml, 1 mg / ml, 1.5 mg / ml) After the addition of 50 μl of CCK-8 assay solution, the reaction was carried out at 37 ° C in a
멜라닌을 합성하는 세포주 B16-F10 세포주에 대한 세포독성Cytotoxicity to cell line B16-F10 cell line that synthesizes melanin
먼저 배양한 세포에 농도별(0.1mg/ml, 0.5mg/ml, 1mg/ml, 1.5mg/ml) 청매실 추출물, 청매실 발효추출물을 처리 후 24시간 경과 후 CCK-8 assay solution을 50ul씩 넣고 37℃ CO2 인큐베이터(incubator)에서 3시간 반응 후 OD450nm의 파장에서 OD값을 확인한다. 또한 네거티브 콘트롤(negative control)로 세포와 CCK-8 solution과 반응시킨 OD값과 소재 및 CCK-8 assay solution이 들어간(소재별 농도별로) OD값을 blank값으로 사용하여 실험하였으며 그 결과는 도 2와 같다.
After 24 hours, 50 μl of CCK-8 assay solution was added to the cultured cells, and the cells were incubated for 1 hour at 37 ° C. After incubating for 3 hours in a CO2 incubator, check the OD value at the OD450nm wavelength. The OD value and the OD value of the material and the CCK-8 assay solution (by concentration) were used as blank values in a negative control, and the results were shown in FIG. 2 .
(실시예 5) 청매실 추출물, 청매실 발효추출물의 멜라닌 세포에서 멜라닌 형성에 미치는 영향 (Example 5) cheongmaesil extract, cheongmaesil effect on melanin formation in melanocytes in the fermentation extract
티로시나제는 시그마(Sigma)사에서 구입하여 사용하였고, 기질인 L-티로신(Sigma)은 0.05M 인산나트륨 완충용액(pH 6.8)에 용해하여 0.1mg/ml 용액으로 만들어 사용하였다. 청매실 추출물, 청매실 발효 추출물은 0.05M 인산나트륨 완충용액(pH 6.8)에 농도별 (25, 50, 100, 500, 1000, 1,500 ug/ml) 조절하여 용해한 후 사용하였다. L-티로신 용액 0.5ml를 시험관에 넣고 여기에 청매실 추출물, 발효 추출물 농도별(25, 50, 100, 500, 1000, 1,500 ug/ml) 0.5ml을 가하고 37℃ 항온기에서 10분간 방치하였다.
Tyrosinase was purchased from Sigma, and the substrate, L-tyrosine (Sigma), was dissolved in 0.05 M sodium phosphate buffer (pH 6.8) and used as a 0.1 mg / ml solution. The fermented extracts of Cheuremil chamber and Cheongmil chamber were prepared by dissolving in 0.05 M sodium phosphate buffer solution (pH 6.8) by concentration (25, 50, 100, 500, 1000, 1,500 ug / ml) 0.5 ml of L-tyrosine solution was added to a test tube, 0.5 ml of each of the browning room extract, fermented extract concentration (25, 50, 100, 500, 1000, 1,500 ug / ml) was added and left for 10 minutes at 37 ° C.
그후, 시료액에 200 unit/ml 티로시나제 0.5ml을 첨가하고 37℃에서 10분간 반응시켰다. 이 반응액이 든 시험관을 얼음 위에 놓아 급냉시켜 반응을 중지시킨 후, 분광광도계를 파장 475nm에서의 흡광도를 측정하였다.
Thereafter, 0.5 ml of 200 units / ml tyrosinase was added to the sample solution, and the mixture was reacted at 37 ° C for 10 minutes. The reaction tube was placed on ice to quench the reaction, and the absorbance of the reaction tube was measured with a spectrophotometer at a wavelength of 475 nm.
음성대조군으로는 상기 시료 대신 완충용액 0.5ml를 넣은 것을 사용하였고 양성 대조군으로는 미백제로 잘 알려진 알부틴을 사용하였다.
As a negative control, 0.5 ml of a buffer solution was used instead of the above sample, and arbutin, well known as a whitening agent, was used as a positive control.
상기 각 시료의 티로시나아제 활성 저해율은 수학식 1에 따라 계산하였다.The inhibition rate of tyrosinase activity of each of the above samples was calculated according to Equation (1).
청매실 추출물과 청매실 발효 추출물의 농도별(25, 50, 100, 500, 1000, 1,500 ug/ml)로 처리한 후 멜라닌 세포에서 멜라닌 생성 억제능을 확인 한 결과 최소 농도 50 ug/ml의 농도에서도 양성대조군 알부틴(Arbutin) 보다 미백효능이 우수한 것을 확인하였다(도 3 참조).
The inhibitory effect of melanin on the melanin production was investigated at the concentration of 50 ug / ml at the concentration of 25, 50, 100, 500, 1000, 1,500 ug / ml. It was confirmed that the whitening effect was superior to that of arbutin (see Fig. 3).
(실시예 6) 청매실 발효추출물의 멜라닌 세포에서 멜라닌 형성에 관여하는 단백질 발현 확인 (Example 6) Confirmation of protein expression involved in melanin formation in melanocytes of a fermented extract of Cheongmilan
청매실 발효추출물을 B16F10 뮤린 멜라노싸이트에 농도별로 처리하여 48시간 배양 후 멜라닌 관련 단백질의 발현을 확인하였다. MITF, Tyrosinase, TRP1, TRP2 모두 청매실 발효추출물을 농도별로 처리시 감소하는 것을 확인하였으며 이것은 청매실 발효추출물이 멜라닌 생성에 관여하는 단백질의 발현을 억제함으로써 멜라닌 형성을 억제함을 알 수 있었다.The browning room fermented extract was treated with B16F10 murine melanocyte at different concentrations and cultured for 48 hours to confirm the expression of melanin related proteins. MITF, Tyrosinase, TRP1, and TRP2 decreased in the browning room fermentation extract by concentration, indicating that the browning room fermented extract inhibited melanogenesis by inhibiting the expression of proteins involved in melanogenesis.
또한 a-MSH는 Tyrosinase의 production을 증가시켜 멜라닌 생합성을 증가시킨다고 알려져 있는데 여기서 우리는 매실 초음파추출물과 a-MSH 함께 처리 시 MITF와 Tyrosinase의 발현이 감소시키는 것으로 보아 매실 초음파추출물이 직접적으로 tyrosinase activity를 억제하지 않고 MITF 와 Tyrosinase, TRP1, TRP2 등의 멜라닌 형성 촉진 인자들의 발현을 억제하여 hypopigmentary effect에 영향을 주는 것을 암시한다.
In addition, a-MSH is known to increase melanin biosynthesis by increasing the production of tyrosinase. Here, we found that the expression of MITF and tyrosinase decreased in the treatment of platelet supernatant and a-MSH, suggesting that platelet supernatant extract directly inhibited tyrosinase activity And inhibit the expression of MITF, tyrosinase, TRP1, TRP2, and other melanin promoting factors, thereby affecting the hypopigmentary effect.
청매실 발효추출물은 멜라노젠을 억제하거나 MEK/Erk signaling을 activation(활성화) 시켜 멜라닌 생성인자들을 (MITF, TRP1, TRP2, Tyrosinase) 억제시킨다. 또한 Akt signal을 활성화 시키고 Wnt / b-catenin signal(시그널) 을 억제시켜 b-catenin의 발현양을 감소시켜서 멜라닌 형성 관련인자들의 발현을 억제하게 된다.Fermented extracts from Cheongmil chamber inhibit melanogenesis or activate melanin factors (MITF, TRP1, TRP2, Tyrosinase) by activating MEK / Erk signaling. In addition, it activates the Akt signal and suppresses the Wnt / b-catenin signal (signal), thereby decreasing the expression of b-catenin and inhibiting the expression of melanin-related factors.
결과적으로, 청매실 발효 추출물은 멜라닌형성에 관여하는 인자들을 억제 시킴으로써 멜라닌 합성을 억제시키고 미백효능을 증가시키게 된다는 증거이다(도 4 및 도 5 참조).
As a result, it is evident that the browning room fermentation extract inhibits melanin synthesis by inhibiting factors involved in melanin formation and increases whitening efficacy (see FIGS. 4 and 5).
(실시예 7) 제형예 (화장수 제형) (Example 7) Formulation Example (lotion formulation)
하기 표 3의 조성에서 가용화 부분을 미리 혼합하여 투명하게 용해시킨 다음, 미리 혼합 점증된 수상에 서서히 투입하면서 20rpm 패들 믹서(Paddle Mixer)로 교반하여, 화장수를 제조하였다.In the composition shown in the following Table 3, the solubilized portions were premixed and dissolved in a transparent state, and then the resulting mixture was stirred with a 20 rpm paddle mixer while gradually adding the mixture to the water-increasing aqueous phase to prepare a lotion.
보습제로는 위의 글리세린, 부틸렌글라이콜, 소듐하이알로로네이트 외에, 피이지 계통의 다가알코올류, 각종 당류(솔비톨, 자일리톨, 글루코오스, 만난 등), 우레아 등을 사용할 수 있다.Examples of the humectant include glycerin, butylene glycol, and sodium hyaluronate, as well as phage-based polyhydric alcohols and various saccharides (sorbitol, xylitol, glucose, mannan, etc.) and urea.
점증제로는, 잔탄검 외에, 카보머, 아크릴릭폴리머계, 셀룰로오스유도체, 글루코단백질류(뮤신, 헤파린, 콘드로이틴 설페이트 등), 점토성분계 분산안정점증제(벤토나이트, 마그네슘알루미늄실리케이트 등)을 사용할 수 있다. As the incrementing agent, carbomers, acrylic polymers, cellulose derivatives, glucoproteins (mucin, heparin, chondroitin sulfate and the like) and clay dispersion stabilizers (bentonite, magnesium aluminum silicate, etc.) can be used in addition to xanthan gum.
가용화제로는, 위의 PEG 하이드로제네이트디 캐스터오일 외에 다양한 친수성 계면활성제(HLB 16이상)를 쓸 수 있다.
As the solubilizing agent, various hydrophilic surfactants (HLB 16 or more) can be used in addition to the above PEG hydrogene decanter oil.
(실시예 8) 제형예 (영양크림 또는 에멀젼 제형) (Example 8) Formulation example (nutritional cream or emulsion formulation)
하기 표 4의 조성에서 각각 유상과 수상은 75℃에서 완전 용해시키고, 4,000 rpm에서 5분간 유화시키고 냉각하여 30℃에서 크림 제형을 제조하였다.In the compositions shown in Table 4 below, the oil phase and water phase were completely dissolved at 75 ° C, emulsified at 4,000 rpm for 5 minutes, and cooled to prepare a cream formulation at 30 ° C.
단, 청매실 추출물 또는 청매실 발효 추출물은 45℃에서 투입하였다. However, the Cheol-myeong-chamber extract or the browning chamber fermented extract was added at 45 ° C.
식물성 오일로는, 참기름, 마카다미아넛오일, 올리브유, 동백유, 밀배아유, 유채종자유, 스쿠알란, 호호바유 등을 단독 또는 수종 혼합하여 사용할 수 있으며 오일류의 산패방지를 위해 산화방지성분(BHT 또는 토코페릴아세테이트)를 소량 사용할 수 있다.As the vegetable oil, sesame oil, macadamia nut oil, olive oil, camellia oil, wheat germ oil, rape seed oil, squalane and jojoba oil can be used singly or as a mixture. Acetate) can be used in small amounts.
유화제로는 위의 폴리소르베이트와 소르비탄세스퀴올리에이트 외에, 다양한 비이온성 친수계면활성제+비이온성 친유계면활성제를 조합하여 HLP 10 - 12조 조정하여 그 합을 2 - 4 정도 범위로 사용할 수 있다. As the emulsifier, various nonionic hydrophilic surfactants + nonionic hydrophilic surfactants may be combined with HLP 10 - 12 groups in addition to the above polysorbate and sorbitan sesquioleate, and the sum thereof may be used in the range of about 2 to 4 have.
알칼리제로 트리에탄올아민 외에 포타슘하이드록사이드나 소듐하이드록사이드 등을 사용할 수 있으며, 점증제로 카보머 등을 사용하는 경우, 알지닌을 사용할 수도 있다.
In addition to triethanolamine as an alkali agent, potassium hydroxide, sodium hydroxide, and the like can be used, and when a carbomer or the like is used as an increasing agent, arginine can also be used.
(실시예 9) 제형예 (팩 제형) (Example 9) Formulation Example (Pack Formulation)
하기 표 5의 조성에서 각각 유상과 수상은 75℃에서 완전 용해시키고, 4,000 rpm에서 8분간 유화시키고 냉각하여 30℃에서 팩 제형을 제조하였다.In the compositions shown in the following Table 5, the oil phase and water phase were completely dissolved at 75 ° C, emulsified at 4,000 rpm for 8 minutes, and cooled to prepare a packed formulation at 30 ° C.
단, 청매실 추출물 또는 청매실 발효 추출물은 45℃에서 투입하였다. However, the Cheol-myeong-chamber extract or the browning chamber fermented extract was added at 45 ° C.
식물성 오일로는, 참기름, 마카다미아넛오일, 올리브유, 동백유, 밀배아유, 유채종자유, 스쿠알란, 호호바유 등을 단독 또는 수종 혼합하여 사용할 수 있으며 오일류의 산패방지를 위해 산화방지성분(BHT 또는 토코페릴아세테이트)를 소량 사용할 수 있다.
As the vegetable oil, sesame oil, macadamia nut oil, olive oil, camellia oil, wheat germ oil, rape seed oil, squalane and jojoba oil can be used singly or as a mixture. Acetate) can be used in small amounts.
이상과 같은 본 발명은 일 실시예에 한정되어 설명되었으나, 이에 한정되지 않고 본 발명의 기술적 사상을 토대로 변형되는 실시예들은 모두 본 발명의 권리범위에 속함이 분명하다.While the present invention has been described with reference to exemplary embodiments, it is to be understood that the invention is not limited to the disclosed exemplary embodiments.
Claims (3)
The dried plum powder and water having a weight of about 1 to 10 times the weight of the dry plum powder are extracted with a solvent at an extraction temperature of 20 to 60 캜 for about 12 to 24 hours at a low temperature for 2 to 3 times to obtain an extract After the filtration, the extract was filtered with a 0.2-μm filter. The extract was inoculated with 1 ~ 10% of a culture of Kazachstania unispora and cultured at 20 ~ 25 ℃ for 4 ~ 7 days. And a freeze-dried powder obtained by filtration under reduced pressure and lyophilized and obtained as a freeze-dried powder of a browning chamber fermentation extract, as an active ingredient.
A cosmetic composition for skin whitening comprising a browning room fermented extract prepared by the method of claim 1.
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KR20190036269A (en) * | 2017-09-27 | 2019-04-04 | 대구한의대학교산학협력단 | Japanese apricot essential oil and cosmetic composition for skin whitening containing the same |
CN109806204A (en) * | 2019-04-04 | 2019-05-28 | 烟台新时代健康产业日化有限公司 | A kind of fermenting extraction process of green plum-blossom |
KR102287416B1 (en) * | 2020-03-31 | 2021-08-09 | 주식회사 더가든오브내추럴솔루션 | Fruit extract complex of vitis vinifera, pyrus malus, prunus mume and carica papaya and cosmetic composition containing of the same |
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KR20020043339A (en) * | 2000-12-02 | 2002-06-10 | 이재민 | Cosmetics compositions having a plum extract |
JP2006124290A (en) | 2004-10-27 | 2006-05-18 | Yonezawa Biru System Service:Kk | Microorganism cultured material obtained by culture medium containing mugwort extract as main ingredient and product using the same |
KR20080094459A (en) * | 2007-04-20 | 2008-10-23 | 주식회사 엘지생활건강 | Cosmetic composition having antioxidation activity |
KR101820854B1 (en) * | 2011-01-28 | 2018-01-23 | 에스케이바이오랜드 주식회사 | Cosmetic compositions for skin whitening comprising fermented extracts of roots, leaves and fruits of Prunus mume Siebold & Zucc. |
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KR20190036269A (en) * | 2017-09-27 | 2019-04-04 | 대구한의대학교산학협력단 | Japanese apricot essential oil and cosmetic composition for skin whitening containing the same |
CN109806204A (en) * | 2019-04-04 | 2019-05-28 | 烟台新时代健康产业日化有限公司 | A kind of fermenting extraction process of green plum-blossom |
KR102287416B1 (en) * | 2020-03-31 | 2021-08-09 | 주식회사 더가든오브내추럴솔루션 | Fruit extract complex of vitis vinifera, pyrus malus, prunus mume and carica papaya and cosmetic composition containing of the same |
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