KR101040508B1 - Skin anti-aging and moisturizing cosmetic composition comprising mung bean fermentation-enzyme extract - Google Patents
Skin anti-aging and moisturizing cosmetic composition comprising mung bean fermentation-enzyme extract Download PDFInfo
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- KR101040508B1 KR101040508B1 KR1020080074838A KR20080074838A KR101040508B1 KR 101040508 B1 KR101040508 B1 KR 101040508B1 KR 1020080074838 A KR1020080074838 A KR 1020080074838A KR 20080074838 A KR20080074838 A KR 20080074838A KR 101040508 B1 KR101040508 B1 KR 101040508B1
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- South Korea
- Prior art keywords
- mung bean
- fermentation
- cosmetic composition
- skin
- enzyme extract
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
- A61K8/66—Enzymes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Dermatology (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Cosmetics (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
본 발명은 녹두를 효모(Saccharomyces cerevisiae) 또는 유산균(Lactobacillus)으로 배양하여 얻은 녹두 발효액을 다시 단백질 분해 효소를 처리하여 얻은 녹두 발효-효소 추출액을 유효성분으로 포함하는 피부보호용 화장료 조성물 및 화장 방법에 관한 것이다. 본 발명에 따른 녹두 발효-효소 추출액은 이소플라본 함량이 증가되고, 피부 세포 활성을 증진시키고, 피부 자극을 완화 시켜주며, 콜라겐 등의 피부 세포 단백질의 합성을 촉진시켜 줌으로써 노화로 인해 발생하는 다양한 피부 트러블을 개선하며 피부를 보호해주는 효능을 가지고 있다.The present invention relates to a cosmetic composition and a cosmetic method for protecting skin comprising a mung bean fermentation-enzyme extract obtained by treating a mung bean fermentation broth obtained by culturing mung bean with yeast ( Saccharomyces cerevisiae ) or Lactobacillus as an active ingredient will be. Mung bean fermentation-enzyme extract according to the present invention increases the isoflavone content, promotes skin cell activity, alleviates skin irritation, and promotes the synthesis of skin cell proteins such as collagen. It has the effect of improving trouble and protecting skin.
녹두 발효, 효모, 유산균, 단백질 분해 효소, 피부 보호, 콜라겐 합성, 주름 개선, 피부 보습, 노화 방지 Mung bean fermentation, yeast, lactic acid bacteria, proteolytic enzymes, skin protection, collagen synthesis, wrinkle improvement, skin moisturizing, anti-aging
Description
본 발명은 녹두 발효-효소 추출액을 함유하는 화장료 조성물에 관한 것으로서, 보다 상세하게는 녹두를 효모 또는 유산균으로 배양하여 얻은 발효 추출액을 단백질 분해효소를 이용하여 2차적으로 처리하여 얻은 발효-효소 추출액을 유효성분으로 하는 화장료 조성물에 관한 것이다. The present invention relates to a cosmetic composition containing a mung bean fermentation-enzyme extract, and more particularly, a fermentation-enzyme extract obtained by secondarily treating a fermentation extract obtained by culturing mung bean with yeast or lactic acid bacteria using protease. It relates to a cosmetic composition as an active ingredient.
사람의 피부는 노화가 진행됨에 따라 다양한 노화 현상들을 나타낸다. 일반적인 현상으로는 피부가 건조해지고, 탄력 저하, 주름 생성이 일어나고 피부도 칙칙해진다. 또한 외부의 약한 자극에도 쉽게 민감해지며 회복에도 오랜 시간이 걸린다. 이러한 노화 현상은 시간이 지남에 따라 자연스럽게 일어나는 현상으로 100% 막을 수 없지만 다양한 방법을 통해 노화 현상이 일어나는 것을 더디게 하거나 예방할 수 있으며 대부분의 화장품이 이러한 노화 현상을 예방하는데 중점을 두고 있 다. Human skin exhibits various aging phenomena as it ages. Common phenomena include dry skin, loss of elasticity, wrinkle formation, and dull skin. It is also easily sensitive to external weak stimuli and takes a long time to recover. These aging phenomena occur naturally over time and can not be 100% prevented, but there are many ways to slow down or prevent aging from happening, and most cosmetic products are focused on preventing such aging.
화장품을 이용하여 피부 노화 현상을 억제하기 위한 가장 유용한 방법은 다양한 피부 노화 방지 성분을 화장품에 첨가하여 사용함으로써 예방할 수 있다. 이를 위해 많은 화장품 회사들이 피부 노화 방지 성분을 개발하기 위해 많은 노력을 하고 있으며, 최근에는 발효를 통해 얻은 유효성분을 제품에 적용하기도 한다. The most useful method for suppressing skin aging phenomenon using cosmetics can be prevented by adding various skin anti-aging ingredients to cosmetics. To this end, many cosmetic companies are making great efforts to develop anti-aging ingredients, and recently, active ingredients obtained through fermentation are applied to products.
예를 들면, 청국장 및 낫도(natto)라는 대표적인 발효 식품에 이용되는 고초균에 의한 발효 산물을 이용하여 생산한 발효액을 이용하여 화장료 조성물을 제공하는 경우가 있다. 이러한 발효액은 미생물에 의해 자극성분들이 분해되거나 다른 물질로 전환됨으로써 피부 자극이 저하되는 장점들이 있으며, 미생물에 의한 발효 산물들이 또한 피부에 좋은 영향을 줌으로써 많이 이용되고 있다.For example, a cosmetic composition may be provided using a fermentation broth produced using a fermentation product by Bacillus subtilis used in representative fermented foods such as Cheonggukjang and natto. Such fermentation broth has the advantage that the skin irritation is lowered by the decomposition of irritants by the microorganisms or converted to other substances, and the fermentation products by the microorganisms are also used as a good effect on the skin.
이에, 본 발명자들은 노화로 인해 발생하는 다양한 피부 현상을 해결하기 위하여 예의 노력한 결과, 녹두를 효모 또는 유산균 등을 이용하여 발효시킨 녹두 발효 추출액이 노화예방 효과가 우수하며, 특히, 녹두 발효 추출액을 단백질 분해효소로 2차 처리할 경우 피부 노화로 인해 발생하는 피부의 문제점, 특히 피부건조, 피부 주름 생성 등을 보다 효과적으로 개선할 수 있음을 확인함으로써 본 발명을 완성하였다.Therefore, the present inventors have made efforts to solve various skin phenomena due to aging, Mung bean fermentation extract fermented with mung beans using yeast or lactic acid bacteria is excellent anti-aging effect, in particular, mung bean fermentation extract protein The present invention was completed by confirming that secondary treatment with a degrading enzyme can more effectively improve skin problems caused by skin aging, particularly skin drying and skin wrinkle formation.
따라서 본 발명의 목적은 녹두 발효-효소 추출액을 포함하는 피부 노화 방지용 화장료 조성물을 제공하는 것이다.Accordingly, an object of the present invention is to provide a cosmetic composition for preventing skin aging comprising mung bean fermentation-enzyme extract.
또한, 본 발명의 목적은 녹두 발효-효소 추출액을 포함하는 피부 주름 개선용 화장료 조성물을 제공하는 것이다. It is also an object of the present invention to provide a cosmetic composition for improving skin wrinkles comprising mung bean fermentation-enzyme extract.
또한, 본 발명의 목적은 녹두 발효-효소 추출액을 포함하는 피부 보습용 화장료 조성물을 제공하는 것이다.It is also an object of the present invention to provide a skin moisturizing cosmetic composition comprising mung bean fermentation-enzyme extract.
또한, 본 발명의 다른 목적은, 녹두 발효-효소 추출액을 포함하는 화장료 조 성물을 피부에 도포하여 콜라겐의 생합성을 촉진 효과, 피부세포를 활성을 촉진 효과 및 피부 보습효과를 나타내는 것을 특징으로 하는 화장 방법을 제공하는 것이다.Another object of the present invention is to apply a cosmetic composition comprising mung bean fermentation-enzyme extract to the skin to promote collagen biosynthesis, skin cell activity promoting effect and skin moisturizing effect To provide a way.
본 발명의 다른 목적 및 이점은 하기의 실시예 및 청구범위에 의해 보다 명확하게 된다.Other objects and advantages of the present invention will become apparent from the following examples and claims.
본 발명은 녹두 발효-효소 추출액을 포함하는 피부 노화 방지용 화장료 조성물에 관한 것이다. 본 발명의 화장료 조성물에 포함된 유효성분인 녹두 발효-효소 추출액은 녹두를 효모 또는 유산균으로 배양하여 수득한 발효액을 다시 단백질 분해 효소를 이용하여 2차 처리해 줌으로써 획득될 수 있으며, 이 추출액은 세포 활성 및 콜라겐 합성을 촉진해주며, 세포 자극 완화 효과가 우수하며, 이를 함유하는 화장료의 경우, 우수한 주름 개선 효과 및 피부 보습효과를 가지고 있어, 이를 화장료 소재로 개발할 경우 탁월한 피부 노화 방지 효과를 기대할 수 있다. The present invention relates to a cosmetic composition for preventing skin aging comprising mung bean fermentation-enzyme extract. Mung bean fermentation-enzyme extract, which is an active ingredient included in the cosmetic composition of the present invention, may be obtained by subjecting the fermentation broth obtained by culturing mung beans with yeast or lactic acid bacteria to secondary treatment with proteolytic enzymes. And it promotes collagen synthesis, excellent cell stimulation effect, cosmetics containing it has excellent wrinkle improvement and skin moisturizing effect, when developing it as a cosmetic material can be expected to excellent skin anti-aging effect .
상기 목적을 달성하기 위해, 본 발명은 녹두 발효-효소 추출액을 포함하는 피부 노화 방지용 화장료 조성물을 제공한다.In order to achieve the above object, the present invention provides a cosmetic composition for preventing skin aging comprising mung bean fermentation-enzyme extract.
또한, 본 발명은 녹두 발효-효소 추출액을 포함하는 피부 주름 개선용 및 피 부 보습용 화장료 조성물을 제공한다. The present invention also provides a cosmetic composition for improving skin wrinkles and skin moisturizing, including mung bean fermentation-enzyme extract.
본 발명의 조성물에서 유효성분으로 이용되는 녹두(Phaseolus radiatus, Mung bean)는 안두 또는 길두라고도 하며, 콩과의 한해살이풀 녹두의 종자로서, 따뜻한 기후의 양토(壤土:모래와 점토가 알맞게 섞인 검은빛의 흙)에서 잘 자라며, 높이는 30∼80cm이다. 줄기는 가늘고 세로로 난 맥이 있고 10여 개의 마디가 있으며 가지를 친다. 잎은 1쌍의 떡잎과 갓 생겨난 잎이 나온 뒤, 3개의 작은 겹잎이 나온다. Mung bean ( Phaseolus radiatus, Mung bean) used as an active ingredient in the composition of the present invention, also known as andu or gildu, is a seed of perennial green mung beans, which is a warm mixture of loam (모: sand and clay of warm climate). It grows well in the soil, and its height is 30 ~ 80cm. The stem has a thin, vertical vein, about 10 nodes, and branches. The leaves come in a pair of cotyledons and freshly formed leaves, followed by three small double leaves.
이러한 녹두는 콩과의 다른 콩류와는 달리, 한방에서 피부병 치료, 해열 및 해독제로 쓰여 왔을 뿐만 아니라, 동의보감에서는 녹두분으로 만들어 얼굴에 발랐을 때 미용효과를 얻을 수 있다고 알려져 왔다. 최근까지도 녹두는 건조 분말의 형태로 또는 이를 물이나 꿀에 개어 얼굴 마사지에 사용되거나, 또는 녹두가루를 물에 섞어 만드는 녹두전과 녹두로부터 전분을 뽑아 만드는 청포묵, 빈대떡, 녹두죽, 숙주나물 등의 식품으로서 예부터 사용되어 왔을 뿐이다.Unlike other legumes such as legumes, such green beans have been used as a treatment for skin diseases, antipyretic and antidote in oriental medicine, and in synonymous gagam, it has been known that beauty effects can be obtained when it is made with green beans. Until recently, mung bean is used as a dry powder or as a food for facial massage by dipping it in water or honey, or as a food such as green bean paste, mung bean cake, mung bean porridge, mung bean sprouts, mung bean sprouts made by mixing mung bean powder with water. It has only been used since ancient times.
본 명세서에서 "녹두"는, 녹두의 다양한 기관(예: 씨앗(열매)잎, 꽃, 줄기 및 뿌리 등)을 의미하며, 바람직하게는 녹두의 씨앗(열매)을 의미한다.As used herein, “mung beans” refers to various organs of mung beans (eg, seeds (berry) leaves, flowers, stems, roots, etc.), and preferably refers to seeds (berries) of mung beans.
한편, 본 명세서에서 용어, 녹두 발효-효소 추출액은 녹두 추출물을 발효균으로 발효시시켜 1차적으로 녹두 발효 추출액을 얻은 후 이를 다시 단백질 분해효 소를 처리하여 얻어지는 추출액을 의미하는 것으로, 발효균의 종류에 따라 녹두 효모 발효-효소 추출액 또는 녹두 유산균 발효-효소 추출액으로 나뉜다. 여기에서 녹두 추출물의 발효균은 효모와 유산균이 주로 사용되며, 발효균으로 효모를 사용할 경우를 녹두 효모 발효-효소 추출액, 발효균으로 유산균을 사용할 경우를 녹두 유산균 발효-효소 추출액으로 이해할 수 있다.Meanwhile, as used herein, the term "mung bean fermentation-enzyme extract" refers to an extract obtained by fermenting mung bean extract with fermentation bacteria, firstly obtaining mung bean fermentation extract, and then treating the protein degradation enzyme again, depending on the type of fermentation bacteria. Therefore, it is divided into mung bean yeast fermentation-enzyme extract or mung bean lactic acid bacteria fermentation-enzyme extract. Here, the fermented bacteria of mung bean extract is mainly used yeast and lactic acid bacteria, and the case of using yeast as fermentation bacteria can be understood as the case of mung bean yeast fermentation-enzyme extract, the case of using lactic acid bacteria as fermentation bacteria.
또한 이의 반대되는 용어로서, 녹두 효소-발효 추출액은 상기 녹두 발효-효소 추출액의 제조과정에서 발효단계와 효소처리단계의 처리를 역순으로 실시하여 얻어지는 추출액을 의미하는 것으로, 녹두 추출물을 우선 단백질 분해효소로 처리한 후 발효균으로 발효시켜 얻은 추출액을 이른다.In addition, as the opposite term, mung bean enzyme-fermented extract means an extract obtained by performing the fermentation step and the enzyme treatment step in the reverse order in the manufacturing process of the mung bean fermentation-enzyme extract, mung bean extract is first proteolytic enzyme After the treatment to fermentation with fermented bacteria to obtain an extract.
본 발명의 녹두를 발효시키는데 사용되는 효모는, 진핵미생물(핵과 세포질이 핵막에 의하여 나누어져 있는 생물)에 속하며, 세포벽, 세포막, 핵, 사립체, 과립 등 세포 소기관을 갖추고 있다. 성의 구별이 없는 이들은 주로 budding(출아법)으로 번식하는데, 분리된 많은 효모들 중에는 발효능이 없는 것, budding 대신 fission(분열) 혹은 bud-fission 방법으로 증식하는 것, 자낭포자(ascospore)가 아닌 사출포자(ballistospore)를 형성하거나 포자(spore)형성이 관찰되지 않는 것 등 다양한 그룹의 효모도 존재한다. 크기는 보통 5-10 × 5-12㎛로 세균의 5-10배 크기이며, 적정 pH는 4.5-5.5이나 1-10에서도 생존한다. pH 1에서 12시간 생존하는 효모는 박테리아보다 산에 강하므로 배지의 pH를 3.5~3.8 정도로 맞추면 박테리아의 오염을 어느 정도 억제할 수 있다.The yeast used to ferment the green beans of the present invention belongs to eukaryotic microorganisms (organisms in which the nucleus and cytoplasm are divided by the nuclear membrane) and have cell organelles such as cell walls, cell membranes, nuclei, granules, and granules. Those without sex distinction breed mainly by budding, among which many of the isolated yeasts have no fermentation capacity, multiply by fission or bud-fission instead of budding, and not ascospore. There are also various groups of yeasts, such as forming spores or not forming spores. The size is usually 5-10 × 5-12㎛, 5-10 times the size of bacteria, and the optimal pH survives at 4.5-5.5 but 1-10. Yeasts that survive 12 hours at pH 1 are more acid resistant than bacteria, so adjusting the pH of the medium to about 3.5 to 3.8 can reduce bacterial contamination.
효모는 균종에 따라서 조금씩 차이가 있으나, 대부분 20~25℃에서 잘 자란다. 보통은 50℃이상에서는 사멸한다고 하지만 그 온도 이상에서 합성이 되지 않는 성장요소를 배지에 직접 첨가해 줌으로써 어느 정도는 극복이 가능하다. 사카로마이세스 세레비시애(Saccharomyces cerevisiae)의 경우 40℃이상에서 사멸하지만 배지에 올레산(oleic acid)와 에르고스테롤(ergosterol)을 첨가해 줌으로써 이를 방지할 수 있다. 효모 균체 자체는 단세포 식물로서 단백질과 미량광물질을 다량 함유하고 있다. 본 발명의 녹두를 발효시키는데 사용하기에 바람직한 효모는 사카로마이세스 세레비시애이다.Yeast is slightly different depending on the species, but most grow well at 20 ~ 25 ℃. Usually, it is killed above 50 ℃, but it can be overcome to some extent by adding the growth factor which is not synthesized above the temperature directly to the medium. Saccharomyces cerevisiae ( Saccharomyces cerevisiae) is killed at 40 ℃ or more, but can be prevented by adding oleic acid (oleic acid) and ergosterol to the medium. Yeast cells themselves are single-celled plants that contain large amounts of protein and trace minerals. A preferred yeast for use in fermenting the green beans of the present invention is Saccharomyces cerevisiae.
본 발명에서 녹두를 발효시키는데 사용된 발효균으로 유산균은, 포도당 또는 유당과 같은 탄수화물을 분해하여 유산(젖산)이나 초산과 같은 유기산을 생성하는 인체에 매우 유익한 미생물의 일종이다. 일반적으로, 유산균이 당으로부터 유산을 만드는 것을 발효라 하며, 이러한 발효과정을 거쳐서 발효식품이 만들어 진다. 본 발명에서 사용될 수 있는 유산균은 예컨대, 락토 바실러스, 스트렙토 코커스, 비피도 박테리움, 류코노스톡, 페디오 코커스, 락토 코커스를 포함하며, 바람직하게는 락토 바실러스(Lactobacillus)이다. As the fermentation bacteria used to ferment mung beans in the present invention, lactic acid bacteria are a kind of microorganisms that are very beneficial to the human body to break down carbohydrates such as glucose or lactose to produce organic acids such as lactic acid (lactic acid) or acetic acid. Generally, lactic acid bacteria make lactic acid from sugar is called fermentation, and fermented food is made through this fermentation process. Lactic acid bacteria that can be used in the present invention include, for example, Lactobacillus, Streptococcus, Bifido bacterium, Leukonostock, Pediococcus, Lactococcus, preferably Lactobacillus.
본 발명의 녹두 발효-효소 추출액은 다음의 단계로 제조될 수 있다:Mung bean fermentation-enzyme extract of the present invention can be prepared by the following steps:
a) 녹두를 분말로 만드는 단계;a) powdering green beans;
b) 상기 단계 a)에서 제조된 녹두 분말을 정제수에 희석한 후 효모 또는 유 산균을 이용하여 발효시키는 단계;b) diluting the mung bean powder prepared in step a) in purified water and then fermenting using yeast or lactic acid bacteria;
c) 상기 단계 b)의 발효에 의해 얻은 발효액을 여과하여 단백질 분해효소로 분해하는 단계; 및c) filtering the fermentation broth obtained by the fermentation of step b) into a protease; And
d) 상기 단계 c)의 효소 분해액을 저온 숙성 및 여과하여 녹두 발효-효소 추출액을 제조하는 단계.d) low-temperature aging and filtration of the enzyme digestion solution of step c) to prepare a mung bean fermentation-enzyme extract.
상기 녹두 발효-효소 추출액의 제조 단계에서, 상기 단계 a)의 녹두의 분말화는 100 내지 500 메쉬 크기 이하로 제조될 수 있으며, 이에 한정될 필요는 없다. In the preparation step of the mung bean fermentation-enzyme extract, the powdered mung bean of step a) may be prepared in a size of 100 to 500 mesh or less, but is not limited thereto.
또한 상기 단계 b)의 발효는 녹두 분말과 정제수를 1:10으로 혼합한 후 121℃에서 멸균시킨 후 유산균(Lactobacillus sp) 또는 효모균(Sacharomyces sp)을 접종하여 배양함으로써 달성된다. In addition, the fermentation of step b) is achieved by inoculating lactic acid bacteria (Lactobacillus sp) or yeast (Sacharomyces sp) after incubation at 121 ° C. after mixing green beans powder and purified water at 1:10.
또한, 발효단계에 사용된 효모 또는 유산균은 총 배양액 1L 기준으로, 1 × 104 내지 1 × 105 세포수의 양으로 접종되는 것이 바람직하며, 각 균주의 성상에 따라 통상적인 조건 및 호기적 또는 통상 혐기(anaerobic)적인 조건에서 배양될 수 있다. 또한, 발효균의 배양을 더욱 활성화시키기 위하여 추가적인 탄소원 및/또는 PH 조절제가 첨가될 수도 있다. 이때 적합한 배양조건은 30 - 37℃, pH 5-7의 호기 또는 통상혐기적인 조건에서 약 1 내지 15일간 배양되며, 바람직하게는 1 내지 7일이며, 더욱 바람직하게는 1 내지 5일이며, 가장 바람직하게는 2 내지 4일간 배양된다. In addition, the yeast or lactic acid bacteria used in the fermentation step is preferably inoculated in the amount of 1 × 10 4 to 1 × 10 5 cells on the basis of 1L of the total culture solution, depending on the characteristics of each strain and aerobic or It can usually be cultured in anaerobic conditions. In addition, additional carbon sources and / or PH regulators may be added to further activate the culture of the fermentor. At this time, suitable culture conditions are incubated for about 1 to 15 days in an aerobic or normal anaerobic condition of 30-37 ℃, pH 5-7, preferably 1 to 7 days, more preferably 1 to 5 days, most Preferably it is incubated for 2 to 4 days.
상기 단계 c)는 단계 b)에서 얻은 발효액을 1차 여과한 후 그 여과액을 단백질 분해효소로 분해하는 단계로서, 이때 여과는 0.25 내지 0.45μm 크기의 여과막으로 여과하여, 여분의 녹두분말알갱이 및 발효균을 제거할 수 있다. 1차 여과막으로 분리된 여과액은 단백질 분해효소로 분해하여 효모 또는 유산균에 의한 발효로도 분해 또는 전환되지 않은 성분들을 2차적으로 분해 또는 전환시키기 위해 실시된다. 이때, 사용되는 단백질 분해효소로는 펩신, 트립신, 카르복시펩티다아제, 아미노펩티다아제, 디펩티다아제 등이 있으며, 바람직하게는 펩신, 트립신, 카르복시펩티다아제이며, 가장 바람직하게는 펩신이다. 상기에서 단계 d)는 단백질 분해효소에 의한 생물전환 방법에 의해 얻어진 추출액을 저온 숙성하여 안정화 시킬 수 있으며, 1차 여과막에 비해 더 세밀한 여과막으로 2차 여과한 후 농축하여 보관할 수 있으며, 이 농축물을 적절히 희석하여 사용한다. 이때 상기 분해-여과-농축의 과정은 보다 유효한 효과가 있는 녹두 발효-효소 추출액을 얻기 위한 공정단계로서, 저온 숙성과정은 5 내지 10일 동안 이루어지는 것이 바람직하며, 여과막의 크기는 0.25μm 이하인 것이 바람직하다.The step c) is a step of firstly filtering the fermentation broth obtained in step b) and then decomposing the filtrate with proteolytic enzymes, wherein the filtration is filtered through a 0.25 to 0.45 μm filtration membrane, and the excess mung bean powder and Fermentation bacteria can be removed. The filtrate separated by the primary filtration membrane is subjected to secondary decomposition or conversion of components that are not degraded or converted even by fermentation by yeast or lactic acid bacteria by proteolytic enzymes. At this time, proteases used are pepsin, trypsin, carboxypeptidase, aminopeptidase, dipeptidase, and the like, preferably pepsin, trypsin, carboxypeptidase, and most preferably pepsin. In step d), the extract obtained by the bioconversion method by proteolytic enzymes can be stabilized by aging at low temperature, and can be stored after the second filtration with a finer filtration membrane than the primary filtration membrane. Properly dilute and use. At this time, the decomposition-filtration-concentration process is a process step for obtaining a more effective mung bean fermentation-enzyme extract, it is preferable that the low temperature ripening process is performed for 5 to 10 days, the size of the filtration membrane is 0.25μm or less. Do.
보다 바람직한 구현예에 따르면, 녹두 분말을 300 메쉬 크기 이하로 분말화한 뒤 배양액에 10 g/L 첨가하고, 효모 균주 또는 유산균 등을 50,000 cfu/L로 첨가하여 각각 효모는 30℃, 유산균은 37℃에서, pH 5-7, 바람직하게는 약 pH 5.5로 호기적 또는 통성 혐기적인 조건으로 발효조에서 약 1일 내지 7일간 배양하여 발효시킨다. 배양 후 상기 발효액은 0.45μm의 여과막을 이용하여 1차 여과한 다음 이 여과액에 다시 단백질 분해효소를 처리한다. 이 처리액은 다시 10일간 저온 숙성 시킨 후 0.25μm 막으로 2차 여과하여 농축한 뒤 본 발명의 녹두 발효-효소 추출액을 제조한다. 사용하기 전에 최종 고형분의 농도를 10g/L가 되게 유지한다.According to a more preferred embodiment, the mung bean powder is powdered to 300 mesh or less and then added to the culture medium 10 g / L, yeast strain or lactic acid bacteria, etc. at 50,000 cfu / L, respectively, yeast 30 ℃, lactic acid bacteria 37 Incubate at pH 5-7, preferably about pH 5.5, incubated in a fermenter for about 1 to 7 days under aerobic or anaerobic conditions. After incubation, the fermentation broth is first filtered using a 0.45 μm filtration membrane, and then the filtrate is subjected to protease again. The treated solution was further aged for 10 days at low temperature, concentrated by secondary filtration with a 0.25 μm membrane, and then prepared mung bean fermentation-enzyme extract of the present invention. The final solids concentration is kept to 10 g / L before use.
이렇게 제조된 본 발명의 녹두 발효-효소 추출액은 이소플라본 함량의 증가(실험예 1), 콜라겐 생합성의 증진 (실험예 2), 세포 활성 촉진 (실험예 3) 및 피부 세포 자극 완화 (실험예 4) 효과가 있으며, 이를 함유하는 화장료 조성물은 주름개선 효과 (실험예 5) 및 피부 보습을 유지에 탁월한 효능(실험예 6)이 있다. 따라서, 본 발명의 녹두 발효-효소 추출액을 화장료 소재로 개발하여 피부 노화를 방지 효과를 기대할 수 있다.The mung bean fermentation-enzyme extract of the present invention prepared in this way is increased in isoflavone content (Experimental Example 1), collagen biosynthesis (Experimental Example 2), cell activity promotion (Experimental Example 3) and skin cell stimulation (Experimental Example 4) ), The cosmetic composition containing the same has an excellent effect (test example 6) to maintain the wrinkles effect (Experimental Example 5) and skin moisturizing. Therefore, the development of mung bean fermentation-enzyme extract of the present invention as a cosmetic material can be expected to prevent skin aging.
본 발명에서, 녹두 발효-효소 추출액의 함량은 전체 화장료 조성물에 대하여 0.0001 - 30.0 %(v/v)이고, 보다 바람직하게는 0.001-20%(v/v)이다. 이때, 녹두 발효-효소 추출액의 함량이 0.0001 %(v/v) 미만일 경우에는 뚜렷한 피부주름 개선 효과 및 피부 보호 효과를 기대할 수 없고, 녹두 발효-효소 추출액의 함량이 30.0%(v/v)를 초과하는 경우에는 함유량 증가에 따른 뚜렷한 효과의 증가가 나타나지 않으며, 피부자극을 유발하거나 제형의 안정성에 있어서 문제가 생긴다.In the present invention, the content of mung bean fermentation-enzyme extract is 0.0001-30.0% (v / v), more preferably 0.001-20% (v / v) based on the total cosmetic composition. In this case, when the content of mung bean fermentation-enzyme extract is less than 0.0001% (v / v), a clear skin wrinkle improvement and skin protection effect cannot be expected, and the content of mung bean fermentation-enzyme extract is 30.0% (v / v). If it is exceeded, there is no apparent effect increase with increasing content, causing skin irritation or problems in the stability of the formulation.
본 발명의 녹두 발효-효소 추출액은 피부에 국소적으로 바르는 거나 뿌리는 것에 의해 그 효과를 달성할 수 있다. 따라서, 본 발명의 바람직한 일 양태에 따르 면, 본 발명의 조성물은 크림, 로션 및 유연수 등의 화장료 조성물 또는 스프레이 및 연고 등의 약제학적 조성물 형태로 제조될 수 있다.Mung bean fermentation-enzyme extract of the present invention can achieve the effect by topical application or spraying on the skin. Therefore, according to one preferred aspect of the present invention, the composition of the present invention may be prepared in the form of cosmetic compositions such as creams, lotions and softening water or pharmaceutical compositions such as sprays and ointments.
본 발명의 화장품 조성물에 포함되는 성분은 유효 성분으로서의 녹두 발효-효소 추출액 이외에 화장품 조성물에 통상적으로 이용되는 성분들을 포함하며, 예컨대 항산화제, 안정화제, 용해화제, 비타민, 안료 및 향료와 같은 통상적인 보조제, 그리고 담체를 포함한다.The components included in the cosmetic composition of the present invention include components conventionally used in cosmetic compositions in addition to the mung bean fermentation-enzyme extract as an active ingredient, and include, for example, conventional agents such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavorings. Adjuvants, and carriers.
본 발명의 화장료 조성물은 당 업계에서 통상적으로 제조되는 어떠한 제형으로도 제조될 수 있으며, 예를 들어, 용액, 현탁액, 유탁액, 페이스트, 겔, 크림, 로션, 파우더, 비누, 계면활성제-함유 클렌징, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션 및 스프레이 등으로 제형 화 될 수 있으나, 이에 한정되는 것은 아니다. 보다 상세하게는, 유연 화장수(스킨), 영양 화장수(밀크로션), 영양 크림, 마사지 크림, 에센스, 아이 크림, 클렌징크림, 클렌징 폼, 클렌징 워터, 팩, 스프레이 또는 파우더의 제형으로 제조될 수 있다.The cosmetic composition of the present invention may be prepared in any formulation commonly prepared in the art, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing , Oils, powder foundations, emulsion foundations, wax foundations and sprays, etc. may be formulated, but is not limited thereto. More specifically, it may be prepared in the form of a flexible lotion (skin), nutrition lotion (milk lotion), nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder. .
본 발명의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물성유, 식물성유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the formulation of the present invention is a paste, cream or gel, animal oils, vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components. Can be.
본 발명의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄 또는 디메틸 에테르와 같은 추진체를 포함할 수 있다.In the case where the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. Especially, in the case of a spray, a mixture of chlorofluorohydrocarbons, propane / Propane or dimethyl ether.
본 발명의 제형이 용액 또는 유탁액인 경우에는 담체 성분으로서 용매, 용해화제 또는 유탁화제가 이용되고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌 글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르가 있다.When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.
본 발명의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상의 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 또는 트라칸트 등이 이용될 수 있다.When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystals Soluble cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.
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, 식물성 유, 라놀린 유도체 또는 에톡실화 글리세롤 지방산 에스테르 등이 이용될 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, an 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 or ethoxylated glycerol fatty acid esters and the like can be used.
또한, 본 발명은 녹두 발효-효소 추출액을 포함하는 화장료 조성물을 인간의 피부에 도포하여 피부 노화로 인해 손상된 다양한 피부 트러블을 개선(세포활성 촉진, 콜라겐 생합성 증진, 피부세포 자극 완화, 피부 보습 유지 등)하는 것을 특징으로 하는 화장방법을 제공한다.In addition, the present invention by applying a cosmetic composition comprising mung bean fermentation-enzyme extract to human skin to improve various skin problems damaged by skin aging (promoting cell activity, collagen biosynthesis, alleviating skin cell stimulation, maintaining skin moisturizing, etc.) It provides a makeup method characterized in that.
본 발명의 화장 방법은 본 발명의 화장료 조성물을 인간의 피부에 도포하는 모든 화장 방법을 일컫는다. 즉, 화장료 조성물을 피부에 도포하는 당업계에 공지된 모든 방법이 본 발명의 화장 방법에 속한다.The cosmetic method of the present invention refers to all cosmetic methods for applying the cosmetic composition of the present invention to human skin. That is, all the methods known in the art for applying the cosmetic composition to the skin belong to the cosmetic method of the present invention.
본 발명의 화장료 조성물은 단독 또는 중복 도포하여 사용하거나, 본 발명 이외의 다른 화장료 조성물과 중복 도포하여 사용할 수 있다. 또한 본 발명에 따른 피부 보호 효과가 우수한 화장료 조성물은 통상적인 사용방법에 따라 사용될 수 있으며, 사용자의 피부 상태 또는 취향에 따라 그 사용횟수를 달리할 수 있다.The cosmetic composition of the present invention may be used alone or in combination, or may be used by overlapping with other cosmetic compositions other than the present invention. In addition, the cosmetic composition with excellent skin protection effect according to the present invention can be used according to a conventional method of use, the number of times of use can be varied according to the user's skin condition or taste.
본 발명의 화장료 조성물이 비누, 계면활성제 함유 클렌징 또는 계면활성제 비 함유 클렌징 제형일 경우, 피부에 도포한 후 닦아내거나 떼거나 물로 씻어낼 수도 있다. 구체적인 예로서, 상기 비누는 액상비누, 가루비누, 고형비누 및 오일비누이며, 상기 계면활성제 함유 클렌징 제형은 클렌징 폼, 클렌징 워터, 클렌징 수 건 및 클렌징 팩이며, 상기 계면활성제 비 함유 클렌징 제형은 클렌징크림, 클렌징 로션, 클렌징 워터 및 클렌징 겔이며, 이에 한정되는 것은 아니다.When the cosmetic composition of the present invention is a soap, a surfactant-containing cleansing or a surfactant-free cleansing formulation, it may be wiped off, peeled off or washed with water after application to the skin. As a specific example, the soaps are liquid soaps, powdered soaps, solid soaps and oil soaps, the surfactant-containing cleansing formulations are cleansing foams, cleansing water, cleansing towels and cleansing packs, and the surfactant-free cleansing formulations are cleansing. Creams, cleansing lotions, cleansing water, and cleansing gels.
본 발명의 녹두 발효-효소 추출액을 포함하는 화장료 조성물을 인간의 피부에 도포하는 화장방법을 수행하면, 피부 노화 예방, 피부 주름개선 및 피부 보습 효과를 효과적으로 개선할 수 있다.When the cosmetic method including the mung bean fermentation-enzyme extract of the present invention is applied to human skin, it is possible to effectively improve skin aging prevention, skin wrinkle improvement and skin moisturizing effect.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 요지에 따라 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .
비교 제조예 1. 녹두 추출물 제조Comparative Preparation Example 1. Preparation of Mung Bean Extract
녹두를 정제수로 세척하고 건조시켜서, 분말화 한 후 100g을 70% 에탄올 용액 0.6L에 첨가하고, 냉각콘덴서가 장착된 환류 추출기에서 4시간 동안 가열 추출하였다. 그런 다음, 수득한 추출액을 300 메쉬 여과포로 여과하고, 와트만 5번 여과지로 여과한 후 감압농축기를 이용하여 용매를 제거하고 동결 건조하여 건조 중량 5.5g을 얻었다. 이렇게 얻은 추출물을 이하 '녹두 추출물'이라 통칭하며, 녹두 추출물은 최종 농도가 1g/100mL되게 사용하였다. Mung beans were washed with purified water, dried, powdered, and then 100 g were added to 0.6 L of a 70% ethanol solution and heated and extracted for 4 hours in a reflux extractor equipped with a cooling capacitor. Thereafter, the obtained extract was filtered through a 300 mesh filter cloth, filtered through Whatman No. 5 filter paper, and the solvent was removed using a vacuum condenser and freeze dried to obtain a dry weight of 5.5 g. The extract thus obtained is referred to hereinafter as 'mung bean extract', and the mung bean extract was used to have a final concentration of 1g / 100mL.
비교 제조예 2. 효모를 이용한 녹두 효모 발효액 제조Comparative Preparation Example 2 Preparation of Mung Bean Yeast Fermentation Solution Using Yeast
정제수로 세척하고 건조시킨 녹두를 분말화 한 후 300메시를 이용하여 미세하게 만들었다. 녹두와 정제수를 1:10으로 혼합하여 121℃에서 멸균시킨 후 효모를 접종하였다. 이때 사용한 효모는 사카로마이세스 새리비시애(Sacharomyces cerevisiae)를 사용하였으며, 배양액 당 50,000 cfu/L로 첨가하였다. 배양은 5L 발효조를 이용하여 3일간, 30℃도, pH 5.5로 유지하며 배양하였다. 배양 후 배양액을 원심 분리하여 배양균을 1차 제거 한 후 0.25 μm 여과막을 이용하여 최종 여과하였다. 이렇게 얻은 여과액을 농축하여 얻은 액을 적정 용매를 이용하여 희석한 후 본 발명에 사용하였다. 이렇게 제조한 발효액을 이하 '녹두 효모 발효액'이라 통칭한다. 이하 실험예에 사용된 녹두 효모 발효액은 최종농도가 1g/100mL가 되게 유지하여 사용하였다.After washing with purified water and dried mung bean powder was made fine by using 300 mesh. Mung beans and purified water were mixed at 1:10 and sterilized at 121 ° C., and then inoculated with yeast. The yeast used at this time was Saccharomyces cerevisiae (Sacharomyces cerevisiae) was used, and was added at 50,000 cfu / L per culture. Cultivation was carried out using a 5L fermenter for 3 days, maintained at 30 ℃, pH 5.5. After cultivation, the culture solution was centrifuged to remove the culture cells first, and then filtered through a 0.25 μm filter membrane. The filtrate thus obtained was concentrated and diluted with a suitable solvent, and used in the present invention. The fermentation broth prepared in this way is hereinafter referred to as ' mung bean yeast fermentation broth '. Mung bean yeast fermentation broth used in the following experimental example was used to maintain a final concentration of 1g / 100mL.
비교 제조예 3. 락토바실러스(Lactobacillus) 속 균주를 이용한 녹두 유산균 발효액 제조Comparative Preparation Example 3 Preparation of Mung Bean Lactic Acid Bacteria Fermentation Solution Using Lactobacillus Species
정제수로 세척하고 건조한 녹두를 분말화 한 후 300메시를 이용하여 미세하게 만들었다. 녹두와 정제수를 1:10으로 혼합하여 121℃에서 멸균시킨 후 유산균을 접종하였다. 이때 사용한 유산균은 락토바실러스(Lactobacillus sp)를 사용하였으며, 배양액 당 50,000 cfu/L로 첨가하였다. 배양은 5L 발효조를 이용하여 3일간, 37℃도, pH 5.5로 유지하며 배양하였다. 배양 후 배양액을 원심 분리하여 배양균을 1차 제거 한 후 0.25 μm 여과막을 이용하여 최종 여과하였다. 이렇게 얻은 여과액 을 농축하여 얻은 액을 적정 용매를 이용하여 본 발명에 사용하였다. 이렇게 제조한 발효액을 이하 '녹두 유산균 발효액'이라 통칭한다. 이하 실험예에 사용된 녹두 유산균 발효액은 최종농도가 1g/100mL가 되게 유지하여 사용하였다. After washing with purified water and dried mung bean powder was made fine by using 300 mesh. Mung beans and purified water were mixed at 1:10 and sterilized at 121 ° C., and then inoculated with lactic acid bacteria. The lactobacillus used was Lactobacillus sp, and was added at 50,000 cfu / L per culture. Cultivation was carried out using a 5L fermenter for 3 days, 37 ℃ ℃, maintained at pH 5.5. After cultivation, the culture solution was centrifuged to remove the culture cells first, and then filtered through a 0.25 μm filter membrane. The filtrate thus obtained was concentrated and used in the present invention using a suitable solvent. The fermentation broth prepared in this way is hereinafter referred to as ' mung bean lactic acid bacteria fermentation broth '. Mung bean lactic acid bacteria fermentation broth used in the following experimental example was used to maintain the final concentration to 1g / 100mL.
비교 제조예 4. 효소 처리 후 효모를 이용한 녹두 효소-효모 발효액 제조Comparative Preparation Example 4 Preparation of Mung Bean Enzyme-Yeast Fermentation Solution Using Yeast after Enzyme Treatment
정제수로 세척하고 건조시킨 녹두를 분말화 한 후 300메시를 이용하여 미세하게 만들었다. 녹두와 정제수를 1:10으로 혼합하여 121℃에서 멸균시킨 후 단백질 분해 효소(펩신, 시그마, 미합중국)를 2시간동안 처리하였다. 처리 후 이 시료에 효모를 접종하였다. 이때 효모는 사카로마이세스 세레비시애(Saccharomyces cerevisiae)를 사용하였다(50,000 cfu/L로 첨가). 배양은 5L 발효조를 이용하여 3일간, 30℃, pH 5.5로 유지하며 배양하였으며, 배양 후 배양액을 원심 분리 및 1차 여과하여 배양균 및 찌꺼기를 제거 한 후, 0.25 μm 여과막을 이용하여 2차 여과 하였다. 이렇게 얻은 여과액을 농축하여 얻은 액을 적정 용매를 이용하여 본 발명에 사용하였다. 이렇게 제조한 발효액을 이하 '녹두 효소-효모 발효액'이라 통칭한다. 이하 실험예에 사용된 녹두 효모 발효액은 최종농도가 1g/100mL가 되게 유지하여 사용하였다.After washing with purified water and dried mung bean powder was made fine by using 300 mesh. Mung beans and purified water were mixed at 1:10 and sterilized at 121 ° C., and then treated with proteolytic enzymes (pepsin, sigma, and the United States) for 2 hours. Yeast was inoculated after treatment. The yeast was used Saccharomyces cerevisiae (added at 50,000 cfu / L). Cultivation was carried out using a 5L fermenter for 3 days, maintained at 30 ° C. and pH 5.5, followed by centrifugation and primary filtration to remove cultures and debris, followed by secondary filtration using a 0.25 μm filter membrane. It was. The solution obtained by concentrating the filtrate thus obtained was used in the present invention using a suitable solvent. The fermentation broth prepared in this way is hereinafter referred to as ' mung bean enzyme-yeast fermentation broth '. Mung bean yeast fermentation broth used in the following experimental example was used to maintain a final concentration of 1g / 100mL.
비교 제조예 5. 효소 처리 후 락토바실러스(Lactobacillus) 속 균주를 이용한 녹두 효소-유산균 발효액 제조Comparative Preparation Example 5 Preparation of Mung Bean Enzyme-Lactic Acid Bacteria Fermentation Solution Using Lactobacillus Strain after Enzyme Treatment
정제수로 세척하고 건조한 녹두를 분말화 한 후 300메시를 이용하여 미세하 게 만들었다. 녹두와 정제수를 1:10으로 혼합하여 121℃에서 멸균시킨 후 단백질 분해 효소(펩신, 시그마, 미합중국)를 2시간동안 처리하였다. 이 시료에 유산균을 접종하여 발효시킨 후 녹두 발효액으로 사용한다. 이때 유산균은 락토바실러스를 사용하였다(50,000 cfu/L로 첨가). 배양은 5L 발효조를 이용하여 3일간, 37℃도, pH 5.5로 유지하며 배양하였다. 배양 후 배양액을 원심 분리 및 1차 여과하여 배양균 및 찌꺼기를 제거 한 후, 0.25 μm 여과막을 이용하여 2차 여과 하였다. 이렇게 얻은 여과액을 농축하여 얻은 액을 적정 용매를 이용하여 본 발명에 사용하였다. 이렇게 제조한 발효액을 이하 '녹두 효소-유산균 발효액'이라 통칭한다. 이하 실험예에 사용된 녹두 유산균 발효액은 최종농도가 1g/100mL가 되게 유지하여 사용하였다. After washing with purified water and powdered dry mung bean was made fine by using 300 mesh. Mung beans and purified water were mixed at 1:10 and sterilized at 121 ° C., and then treated with proteolytic enzymes (pepsin, sigma, and the United States) for 2 hours. This sample is inoculated with lactic acid bacteria and fermented before use as mung bean fermentation broth. At this time, Lactobacillus was used (added at 50,000 cfu / L). Cultivation was carried out using a 5L fermenter for 3 days, 37 ℃ ℃, maintained at pH 5.5. After incubation, the culture solution was centrifuged and primary filtered to remove the culture and debris, and then filtered secondly using a 0.25 μm filtration membrane. The solution obtained by concentrating the filtrate thus obtained was used in the present invention using a suitable solvent. The fermentation broth prepared in this way is hereinafter referred to as ' mung bean enzyme-lactic acid bacterium fermentation broth '. Mung bean lactic acid bacteria fermentation broth used in the following experimental example was used to maintain the final concentration to 1g / 100mL.
제조예 1. 녹두 효모 발효-효소 추출액의 제조Preparation Example 1 Preparation of Mung Bean Yeast Fermentation-Enzyme Extract
정제수로 세척하고 건조시킨 녹두를 분말화 한 후 300메시를 이용하여 미세하게 만들었다. 녹두와 정제수를 1:10으로 혼합하여 121℃에서 멸균시킨 후 효모(Sacharomyces cerevisiae)를 접종하였다. 이때, 효모는 사카로마이세스 세레비시애(Saccharomyces cerevisiae)를 사용하였으며, 배양액 당 50,000 cfu/L로 첨가하였다. 배양은 5L 발효조를 이용하여 3일간, 30℃, pH 5.5로 유지하며 배양하였다. 배양 후 배양액을 원심 분리하여 배양균을 제거 한 후 0.45μm 여과막을 1차 여과 하였다. 이렇게 얻은 여과액에 다시 단백질 분해효소(펩신, 시그마, 미합중국)를 2시간동안 처리한 후 이 액을 0.25μm 여과막을 이용하여 2차 여과하였다. 이 여과액을 다시 농축한 후 적정 용매를 이용하여 희석해서 사용하였다. 이렇게 제조한 발효-효소 추출액을 이하 '녹두 효모 발효-효소 추출액'이라 통칭한다. 이하 실험예에 사용된 녹두 효모 발효 효소액은 최종농도가 1g/100mL가 되게 유지하여 사용하였다.After washing with purified water and dried mung bean powder was made fine by using 300 mesh. Mung beans and purified water were mixed at 1:10 and sterilized at 121 ° C., and then inoculated with yeast (Sacharomyces cerevisiae). At this time, the yeast was used Saccharomyces cerevisiae ( Saccharomyces cerevisiae ), was added at 50,000 cfu / L per culture. Cultivation was carried out using a 5L fermenter for 3 days, maintained at 30 ℃, pH 5.5. After incubation, the culture medium was centrifuged to remove the culture, and then 0.45 μm filtration membrane was first filtered. The filtrate thus obtained was further treated with proteolytic enzymes (pepsin, sigma, and the United States) for 2 hours, and then the solution was filtered secondly using a 0.25 μm filtration membrane. The filtrate was concentrated again and diluted with a suitable solvent. The fermentation-enzyme extract thus prepared is hereinafter referred to as ' mung bean yeast fermentation-enzyme extract '. Mung bean yeast fermentation enzyme solution used in the following experiment was used to maintain a final concentration of 1g / 100mL.
제조예 2. 녹두 유산균 발효-효소 추출액의 제조Preparation Example 2 Preparation of Mung Bean Lactobacillus Fermentation-Enzyme Extract
정제수로 세척하고 건조시킨 녹두를 분말화 한 후 300메시를 이용하여 미세하게 만들었다. 녹두와 정제수를 1:10으로 혼합하여 121℃에서 멸균시킨 후 유산균을 접종하였다. 이때, 유산균으로는 락토바실러스(Lacobacillus sp)를 사용하였으며, 배양액 당 50,000 cfu/L로 첨가하였다. 배양은 5L 발효조를 이용하여 3일간, 37℃, pH 5.5로 유지하며 배양하였다. 배양 후 배양액을 원심 분리하여 배양균을 제거 한 후 0.45μm 여과막을 1차 여과 하였다. 이렇게 얻은 여과액에 다시 단백질 분해효소(펩신, 시그마, 미합중국)를 2시간동안 처리한 후 이 액을 0.25μm 여과막을 이용하여 2차 여과하였다. 이 여과액을 다시 농축한 후 적정 용매를 이용하여 희석해서 사용하였다. 이렇게 제조한 발효-효소 추출액을 이하 '녹두 유산균 발효-효소 추출액'이라 통칭한다. 이하 실험예에 사용된 녹두 효모 발효 효소액은 최종농도가 1g/100㎖가 되게 유지하여 사용하였다After washing with purified water and dried mung bean powder was made fine by using 300 mesh. Mung beans and purified water were mixed at 1:10 and sterilized at 121 ° C., and then inoculated with lactic acid bacteria. At this time, Lactobacillus (Lacobacillus sp) was used, and was added at 50,000 cfu / L per culture. The culture was carried out using a 5L fermenter for 3 days, maintained at 37 ℃, pH 5.5. After incubation, the culture medium was centrifuged to remove the culture, and then 0.45 μm filtration membrane was first filtered. The filtrate thus obtained was further treated with proteolytic enzymes (pepsin, sigma, and the United States) for 2 hours, and then the solution was filtered secondly using a 0.25 μm filtration membrane. The filtrate was concentrated again and diluted with a suitable solvent. The fermentation-enzyme extract thus prepared is hereinafter referred to as ' mung bean lactic acid bacteria fermentation-enzyme extract '. Mung bean yeast fermentation enzyme solution used in the following experimental example was used to maintain a final concentration of 1g / 100ml
실험예 1: 녹두 발효-효소 추출액의 이소플라본 함량 분석Experimental Example 1 Analysis of Isoflavone Contents of Mung Bean Fermentation-Enzyme Extract
이소 플라본 함량을 분석하기 위하여 위 제조예에서 만든 시료들을 고속 액 체 크로마토 그래피(HPLC)를 이용하여 분석하였다. 그 결과는 하기 표 1에 나타내었다. In order to analyze the isoflavone content, the samples prepared in the above preparation were analyzed by high performance liquid chromatography (HPLC). The results are shown in Table 1 below.
제조예1compare
Preparation Example 1
제조예2compare
Preparation Example 2
제조예3compare
Preparation Example 3
제조예4compare
Preparation Example 4
제조예5compare
Preparation Example 5
상기 결과를 볼 때, 이소 플라본류의 함량이 발효 후 효소 처리하여 얻은 본 발명의 제조예 1 및 2의 시료에서 타 시료에 비해 현저하게 증가하는 것을 보여 주었다. In view of the above results, it was shown that the content of isoflavones increased significantly compared to other samples in the samples of Preparation Examples 1 and 2 of the present invention obtained by enzymatic treatment after fermentation.
실험예 2: 녹두 발효-효소 추출액의 콜라겐 생합성 증진 효과 Experimental Example 2: Collagen Biosynthesis Enhancement Effect of Mung Bean Fermentation-Enzyme Extract
인체 정상 섬유아세포를 48-웰 마이크로 플레이트의 각 웰에 1 x 106 세포가 되도록 접종하고, DMEM(Dulbecco's Modified Eagle Medium) 배지에서 37℃에서 24시간 동안 배양하였다. 이어, 비교 제조예 1의 녹두 추출물, 비교 제조예 2의 녹두 효모 발효액, 비교 제조예 3의 녹두 유산균 발효액, 비교 제조예 4의 녹두 효소-효모 발효액, 비교 제조예 5의 녹두 효소-유산균 발효액, 제조예 1의 녹두 효모 발효-효소 추출액 및 제조예 2의 녹두 유산균 발효-효소 추출액을 각각 최종 농도 1.0%로 하여 혈청이 없는 DMEM 배지로 교체한 실험군과 녹두 배양액이 포함되지 않은 혈청이 없는 DMEM 배지로 교체한 대조군을 24시간 동안 추가로 배양하였다. 배양 후, 각 웰의 상층액을 모아 프로콜라겐 (procollagen) 타입 I C-펩타이드 (PICP) 양을 키트 (Takara, 일본)를 이용하여 새로 합성된 콜라겐 양을 측정하였다. PICP 양은 ng/㎖ 환산하였으며, 양성대조군으로는 250uM의 아스코르빈산(ascrobic acid)을 사용하였다. 콜라겐 생합성 증가율은 하기 수학식 1에 따라 계산한 후 그 결과를 표 2에 나타내었다.Human normal fibroblasts were seeded to 1 x 10 6 cells in each well of a 48-well microplate and incubated for 24 hours at 37 ° C. in Dulbecco's Modified Eagle Medium (DMEM) medium. Next, Mung bean extract of Comparative Preparation Example 1, Mung bean yeast fermentation broth of Comparative Preparation Example 2, Mung bean lactic acid bacteria fermentation broth of Comparative Preparation Example 3, Mung bean enzyme-yeast fermentation broth of Comparative Preparation Example 4, Mung bean enzyme-lactic acid bacteria fermentation broth of Comparative Preparation Example 5, Experimental group in which mung bean yeast fermentation-enzyme extract of Preparation Example 1 and mung bean lactic acid bacteria fermentation-enzyme extract of Preparation Example 2 were respectively replaced with DMEM medium without serum and serum-free DMEM medium without mung bean culture solution Controls replaced with were further incubated for 24 hours. After incubation, the supernatant of each well was collected and the amount of newly synthesized collagen was measured using a procollagen type I C-peptide (PICP) kit (Takara, Japan). The amount of PICP was converted to ng / ml, and 250 μM of ascorbic acid was used as a positive control. Collagen biosynthesis growth rate is calculated according to the following equation 1 and the results are shown in Table 2.
이상의 실험예 2를 통해, 본 발명의 녹두 발효-효소 추출액인 제조예 1-2는 녹두 추출물(비교 제조예 1)에 비해 그 효과가 훨씬 탁월했음은 물론, 녹두의 단순 발효액인 녹두 효모 발효액과 녹두 유산균 발효액(비교 제조예 2 및 3) 및 녹두 추출물을 효소 처리 후 발효시킨 녹두 효소-발효액(비교 제조예 4 및 5)에 비해서도 콜라겐 생합성 증진 효과가 우수하였다. 게다가 양성 대조군인 아스코르빈산과 비교시에도 그 효과가 월등하였다.Through the above Experimental Example 2, Preparation Example 1-2 of the Mung bean fermentation-enzyme extract of the present invention was much more effective than the Mung bean extract (Comparative Example 1), as well as mung bean yeast fermentation broth that is a simple fermentation solution of mung beans. Compared with the mung bean lactic acid bacteria fermentation broths (Comparative Preparation Examples 2 and 3) and the mung bean extract fermented after the enzyme treatment, the mung bean enzyme-fermentation solutions (Comparative Preparation Examples 4 and 5) were also superior in collagen biosynthesis. In addition, the effect was superior to that of ascorbic acid, a positive control.
실험예 3: 녹두 발효-효소 추출액의 섬유아세포 증식 효과 Experimental Example 3: Fibroblast Proliferation Effect of Mung Bean Fermentation-Enzyme Extract
인체 정상 섬유아세포를 96-웰 마이크로 플레이트의 각 웰에 1 × 104 세포가 되도록 접종하고, DMEM 배지에서 37℃에서 24시간 동안 배양하였다. 이어, 비교 제조예 1의 녹두 추출물, 비교 제조예 2의 녹두 효모 발효액, 비교 제조예 3의 녹두 유산균 발효액, 비교 제조예 4의 녹두 효소-효모 발효액, 비교 제조예 5의 녹두 효소-유산균 발효액, 제조예 1의 녹두 효모 발효-효소 추출액 및 제조예 2의 녹두 유산균 발효-효소 추출액을 각각 1.0%로 하여 혈청이 없는 DMEM 배지로 교체한 실험군과 녹두 발효-효소 추출액이 포함되지 않은 혈청이 없는 DMEM 배지로 교체한 대조군을 24시간 동안 추가로 배양하였다. 배양 후, 세포의 생존율을 비교하기 위하여 MTT(시그마, 미합중국) 솔루션(3mg/㎖)을 첨가하여 세포 생존율을 ELISA READER(Molecular Devices, 미합중국)를 이용하여 570nm에서 흡광도를 측정하고 하기 수학식 2에 따라 섬유아세포 증식률(%)을 계산하였으며, 실험 결과는 하기 표 3에 기재하였다. 이때 양성 대조군으로 TGF-β를 사용하였다.Human normal fibroblasts were seeded into 1 × 10 4 cells in each well of a 96-well microplate and incubated for 24 hours at 37 ° C. in DMEM medium. Next, Mung bean extract of Comparative Preparation Example 1, Mung bean yeast fermentation broth of Comparative Preparation Example 2, Mung bean lactic acid bacteria fermentation broth of Comparative Preparation Example 3, Mung bean enzyme-yeast fermentation broth of Comparative Preparation Example 4, Mung bean enzyme-lactic acid bacteria fermentation broth of Comparative Preparation Example 5, Experimental group in which Mung bean yeast fermentation-enzyme extract of Preparation Example 1 and mung bean lactic acid bacteria fermentation-enzyme extract of Preparation Example 2 were respectively replaced with DMEM medium without serum and serum-free DMEM without mung bean fermentation-enzyme extract Controls replaced with medium were further incubated for 24 hours. After incubation, MTT (Sigma, US) solution (3 mg / mL) was added to compare cell viability, and cell viability was measured at 570 nm using ELISA READER (Molecular Devices, USA). According to the fibroblast proliferation rate (%) was calculated, the experimental results are shown in Table 3 below. TGF-β was used as a positive control.
상기 표 3의 결과에서도 알 수 있듯이, 본 발명의 녹두 발효-효소 추출액인 제조예 1-2는 녹두 추출물(비교 제조예 1)에 비해 그 효과가 훨씬 탁월했음은 물론, 녹두의 단순 발효액인 녹두 효모 발효액 및 녹두 유산균 발효액(비교 제조예 2 및 3) 및 녹두 추출물을 효소 처리 후 발효시킨 녹두 효소-발효액(비교 제조예 4 및 5)에 비해서도 세포 생성 촉진 효과가 우수하였다. 게다가 양성 대조군인 TGF-β 만큼 우수한 세포 생성 증가율을 보임을 알 수 있었다.As can be seen from the results of Table 3, Preparation Example 1-2 of the mung bean fermentation-enzyme extract of the present invention was much better than the mung bean extract (Comparative Preparation Example 1), as well as mung bean that is a simple fermentation solution of mung beans Yeast fermentation broth, mung bean lactic acid bacteria fermentation broth (Comparative Preparation Examples 2 and 3) and mung bean extract were superior to mung bean enzyme-fermenting solutions (Comparative Preparation Examples 4 and 5) fermented after enzyme treatment. In addition, it can be seen that the cell growth rate is as good as the positive control TGF-β.
실험예 4: 녹두 발효-효소 추출액의 락트산에 의한 세포자극 완화 효과 시험Experimental Example 4: lactic acid mitigation effect test of mung bean fermentation-enzyme extract
인간 피부 세포인 섬유아세포(한국 세포주 은행, 대한민국)를 T-75 플라스크(Falcon, 미합중국)에서 80% 정도 성장할 때까지 배양하였다. 이것을 다시 96-웰 플레이트(Falcon, 미합중국)에 3×104 cells/well이 되게 옮겨 24시간 동안 배양하였다. 배양 후 현미경을 통해 세포가 완전히 부착되어 잘 자라는지 여부를 확인하고 피부세포 자극원으로 락트산을 이용하여 실험을 진행하였다. 이때 락트산은 0.2%를 사용하였다. 즉, 96웰의 각각에 0.2% 락트산이 함유된 DMEM(시그마, 미합중국) 배지 및 락트산 무첨가 DMEM 배지를 200㎕씩 첨가하고, 비교 제조예 1-5의 녹두 추출물, 녹두 효모 발효액 및 녹두 유산균 발효액, 녹두 효소 효모 발효액, 녹두 효소 유산균 발효액, 제조예 1-2의 녹두 발효 효소 추출액을 각각 1.0 %(v/v)로 조절하여 첨가하였다. 이때 락트산, 녹두 발효액 및 녹두 발효-효소 추출액을 모두 첨가하지 않은 것을 대조군으로 하였고, 락트산만 첨가한 것을 비교군으로 하였다. 시험물질을 첨가하고 12시간 경과 후 세포의 생존율을 비교하기 위하여 MTT(시그마, 미합중국) 솔루션(3mg/㎖)을 첨가하여 세포 생존율을 ELISA READER(Molecular Devices, 미합중국)를 이용하여 570nm에서 흡광도를 측정하고 하기 수학식 3에 따라 세포 생존율(%)을 계산하였으며, 실험 결과는 하기 표 4에 기재하였다.Fibroblasts (Korea Cell Line Bank, South Korea), which are human skin cells, were cultured in T-75 flasks (Falcon, United States) until they grew about 80%. This was again transferred to 3 × 10 4 cells / well in 96-well plates (Falcon, USA) and incubated for 24 hours. After culturing, the microscope confirmed whether the cells were completely attached and grew well. The experiment was performed using lactic acid as a source of skin cell stimulation. At this time, lactic acid used 0.2%. That is, 200 μl of DMEM (Sigma, U.S.) medium and lactic acid-free DMEM medium containing 0.2% lactic acid were added to each of the 96 wells, and the mung bean extract, the mung bean yeast fermentation solution, and the mung bean lactic acid fermentation broth of Comparative Preparation Example 1-5, Mung bean enzyme yeast fermentation broth, mung bean enzyme lactobacillus fermentation broth, and mung bean fermentation enzyme extract of Preparation Example 1-2 were adjusted to 1.0% (v / v), respectively. At this time, lactic acid, mung bean fermentation broth and mung bean fermentation-enzyme extract were not added as a control, and only lactic acid was added as a comparison group. In order to compare the viability of the cells 12 hours after the addition of the test substance, the cell viability was measured by adding MTT (Sigma, US) solution (3 mg / mL) at 570 nm using ELISA READER (Molecular Devices, USA). And cell viability (%) was calculated according to the following equation 3, the experimental results are shown in Table 4 below.
상기 표 4의 결과를 통해 알 수 있듯이, 락트산을 처리 및 무처리한 후 본 발명의 녹두 발효-효소 추출액을 처리한 제조예 1-2의 실험결과는, 녹두 추출물을 처리한 비교 제조예 1의 경우에 비해 락트산 처리에 의한 세포 독성을 완화해 주어 세포 생존율을 크게 증가시키는 효과가 탁월했음은 물론, 락트산 처리 및 무처리 후 녹두의 단순 발효액인 녹두 효모 발효액 및 녹두 유산균 발효액을 처리한 비교 제조예 2 과 3 및 녹두 추출물을 효소 처리 후 발효시킨 비교 제조예 4 와 5의 경우에 비해서도 세포 생존율이 훨씬 뛰어나, 세포자극 완화 효과가 뛰어남을 알 수 있었다.As can be seen from the results of Table 4, the experimental results of Preparation Example 1-2, which treated the mung bean fermentation-enzyme extract of the present invention after treating and non-lactic acid treatment, of Comparative Preparation Example 1 treated with mung bean extract Compared to the cases, the cytotoxicity by lactic acid treatment was alleviated and the cell survival rate was greatly increased, and the comparative preparations in which the mung bean yeast fermentation broth and the mung bean lactic acid fermentation broth which were simple fermentation solutions of mung beans after lactic acid treatment and no treatment were treated. Compared to Comparative Preparation Examples 4 and 5 in which 2 and 3 and mung bean extracts were fermented after enzyme treatment, the cell viability was much higher, and the cell stimulating effect was excellent.
이하 상기한 실험예의 결과를 근거로 하여 본 발명의 녹두 발효-효소 추출액을 포함하는 화장료를 조성하여 제시한다. 그러나 본 발명의 조성물을 하기의 처방 예들로 한정하고자 하는 것은 아니다. 하기 처방 예에 사용한 녹두 발효-효소 추출액은 녹두 효모 발효 추출액이나 녹두 유산균 발효 추출액 중 어느 것을 사용하여도 무방하다.Hereinafter, based on the results of the above experimental example, to present a cosmetic composition comprising a mung bean fermentation-enzyme extract of the present invention. However, the composition of the present invention is not intended to be limited to the following formulation examples. The mung bean fermentation-enzyme extract used in the following prescription example may use either a mung bean yeast fermentation extract or a mung bean lactic acid bacteria fermentation extract.
처방예 1: 유연화장수Prescription 1: Softener
녹두 발효-효소 추출액을 함유한 화장료 중 유연화장수의 처방예는 다음과 같다.A prescription example of softening water in cosmetics containing mung bean fermentation-enzyme extract is as follows.
처방예 2: 영양화장수Prescription Example 2: Nutritional Cosmetics
녹두 발효-효소 추출액을 함유한 화장료중 영양화장수의 처방예는 다음과 같다. A prescription example of nutrient cosmetics in cosmetics containing mung bean fermentation-enzyme extract is as follows.
처방예 3: 영양크림Formulation Example 3: Nutritional Cream
녹두 발효-효소 추출액을 함유한 화장료 중 영양크림의 처방예는 다음과 같다. A prescription example of nutrient cream in cosmetics containing mung bean fermentation-enzyme extract is as follows.
실험예 5: 녹두 발효-효소 추출액을 함유한 화장료의 주름 개선 효과 평가Experimental Example 5: Evaluation of wrinkle improvement effect of cosmetics containing mung bean fermentation-enzyme extract
본 발명의 화장료의 주름 개선 효과를 실제 사용 테스트를 통하여 평가하였다. 제조예 2의 녹두 유산균 발효-효소 추출액을 3%(v/v)를 함유하고 있는 처방예 3의 영양 크림과 처방예 3에서 녹두 유산균 발효-효소 추출액을 비교 제조예 5의 녹두 효소-유산균 발효액으로 대체한 크림을 비교 처방예 및 정제수로 대체한 크림을 대조예로 사용하였다. 90명의 여성을 대상으로 얼굴 양쪽면을 사용하여 6주 후의 주름 개선 효과를 육안으로 평가하여 주름 개선 정도를 확인하였다. 이 평가를 토대로 한 주름 개선 효과 결과는 하기의 표 8에 나타내었다.The wrinkle improvement effect of the cosmetics of this invention was evaluated through the actual use test. The nutritional cream of Formula 3, which contains 3% (v / v) of Mung bean lactic acid bacteria fermentation-enzyme extract of Preparation Example 2, was compared to the nutrition cream of Formula 3, and the composition of Mung bean lactic acid bacteria fermentation-enzyme extract, which was prepared in Preparation Example 3, The cream replaced with a comparative prescription and purified water was used as a control. Ninety women were visually evaluated for the improvement of wrinkles after six weeks using both sides of the face to confirm the degree of wrinkle improvement. The wrinkle improvement effect results based on this evaluation are shown in Table 8 below.
상기 표 8의 실험 결과에 의하면, 녹두 발효-효소 추출액을 함유한 본 발명의 처방예 3의 크림 화장료는 녹두 발효-효소 추출액 대신 비교 제조예 5의 녹두 효소-유산균 발효액을 함유하는 비교 처방예 및 녹두 발효-효소 추출액 대신 정세수를 함유하는 대조예에 비하여 아주 높은 주름개선 효과를 보여주었으며, 본 화장료를 피부에 도포한 대부분의 피검자들에게서 피부 자극을 관찰할 수 없었다.According to the results of the experiment of Table 8, the cream cosmetics of Formulation Example 3 of the present invention containing mung bean fermentation-enzyme extract, the comparative formulation example containing mung bean enzyme-lactic acid bacteria fermentation broth of Comparative Preparation Example 5 instead of mung bean fermentation-enzyme extract and Compared with the control example containing the cleanse water instead of the mung bean fermentation-enzyme extract, it showed a very high wrinkle improvement effect, and skin irritation was not observed in most of the subjects who applied the cosmetic to the skin.
실험예 6: 녹두 발효-효소 추출액을 함유한 화장료의 보습 평가Experimental Example 6: Moisturizing Evaluation of Cosmetics Containing Mung Bean Fermentation-Enzyme Extract
본 발명의 화장료의 보습 효과를 실제 사용 테스트를 통하여 평가 하였다. 제조예 2의 녹두 발효-효소 추출액을 3%(v/v)를 함유하고 있는 처방예 3의 영양 크림과 처방예 3에서 녹두 발효-효소 추출액을 비교 제조예 5의 녹두 효소-유산균 발효액으로 대체한 크림을 비교 처방예 및 정제수로 대체한 크림을 대조예로 사용하였다. 30명의 여성을 대상으로 상박에 각각의 제품을 적용할 수 있도록 정 사각형의 제품 적용 부위를 만든 후 2주간 도포해주면서 피부 보습량 변화를 스킨 코네오메타(skin corneometer; C+K courage, 독일)를 이용하여 측정한 후 10%이상 개선된 피검자의 숫자를 하기 표 8에 나타내었으며, 피부 보습 개선율을 계산하였다. 그 결과는 하기의 표 9에 나타내었다.Moisturizing effect of the cosmetic of the present invention was evaluated through a practical use test. Replace the nutrition cream of Formulation Example 3 containing 3% (v / v) of Mung Bean Fermentation-Enzyme Extract of Preparation Example 2 and the Mung Bean Fermentation-Enzyme Extract of Preparation Example 3 with the Mung Bean Enzyme-Lactic Acid Bacteria Fermentation Solution of Preparation Example 5. A cream in which one cream was replaced with a comparative prescription and purified water was used as a control. For 30 women, make a square product area to apply each product to the upper arm and apply it for 2 weeks to apply skin corneometer (C + K courage, Germany) The number of the subjects improved by 10% or more after the measurement was shown in Table 8 below, and the skin moisturization improvement rate was calculated. The results are shown in Table 9 below.
상기 표 9의 실험 결과에서도 알 수 있듯이, 녹두 발효-효소 추출액을 함유한 본 발명의 처방예 3의 크림 화장료는 녹두 발효-효소 추출액 대신 비교 제조예 5의 녹두 효소-유산균 발효액을 함유하는 비교 처방예 및 녹두 발효-효소 추출액 대신 정제수를 함유하는 대조예에 비하여 아주 높은 피부 보습 효과를 보여주었다.As can be seen from the experimental results of Table 9, the cream cosmetics of Formulation Example 3 of the present invention containing mung bean fermentation-enzyme extract is a comparative prescription containing mung bean enzyme-lactic acid bacteria fermentation broth of Comparative Preparation Example 5 instead of mung bean fermentation-enzyme extract Example and showed a very high skin moisturizing effect compared to the control containing purified water instead of mung bean fermentation-enzyme extract.
이상으로 본 발명의 특정한 부분을 상세히 기술하였는바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적인 기술은 단지 바람직한 구현예일 뿐이며, 이에 본 발명의 범위가 제한되는 것이 아닌 점은 명백하다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항과 그의 등가물에 의하여 정의된다고 할 것이다.Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that such a specific technology is only a preferred embodiment, and the scope of the present invention is not limited thereto. Therefore, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.
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| KR102257667B1 (en) * | 2020-04-20 | 2021-05-28 | 그린코스 주식회사 | Cosmetic composition that suppresses the generation of senile body odor and contains a skin moisturizing effect containing coffee bean, mung bean and bamboo shoot complex extract |
| CN115006310B (en) * | 2022-07-05 | 2023-10-24 | 菏泽市亿鑫生物科技有限公司 | Mung bean sprout fermentation product, external skin preparation containing mung bean sprout fermentation product, and preparation method and application of external skin preparation |
| CN115089529B (en) * | 2022-08-03 | 2023-07-25 | 肽源（广州）生物科技有限公司 | Mung bean hull fermentation product with acne removing effect, and preparation method and application thereof |
| CN117281748A (en) * | 2023-07-14 | 2023-12-26 | 广东真丽斯化妆品有限公司 | A cosmetic composition containing mushroom extract for promoting skin repair |
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| US9913799B2 (en) | 2014-07-11 | 2018-03-13 | Mary Kay Inc. | Cosmetic compositions and methods of their use |
| US10123968B2 (en) | 2014-07-11 | 2018-11-13 | Mary Kay Inc. | Cosmetic compositions and methods of their use |
| US10231922B2 (en) | 2014-07-11 | 2019-03-19 | Mary Kay Inc. | Cosmetic compositions and methods of their use |
| US10617633B2 (en) | 2014-07-11 | 2020-04-14 | Mary Kay Inc. | Cosmetic compositions and methods of their use |
| US11969498B2 (en) | 2014-07-11 | 2024-04-30 | Mary Kay Inc. | Cosmetic compositions and methods of their use |
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