KR102167786B1 - manufacturing method of fermented extract of Centella asiatica, Althaea rosea, Blue lotus, Swertia japonica Griseb using Lactobacillus and cosmetic composition using that fermented extract - Google Patents
manufacturing method of fermented extract of Centella asiatica, Althaea rosea, Blue lotus, Swertia japonica Griseb using Lactobacillus and cosmetic composition using that fermented extract Download PDFInfo
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Abstract
Description
본 발명은 화장료 조성물, 화장료용 발효물 및 그 제조 방법에 관한 것으로, 특히 접종 균주로 락토바실러스 쿠르바투스 J2K-01을 사용하고, 발효 대상은 병풀, 접시꽃, 푸른 연꽃, 쓴풀을 사용하여 피부 진정, 상처 치유 및 재생, 피부 염증 개선 효과를 높이도록 한, 락토바실러스를 이용한 병풀, 접시꽃, 푸른연꽃, 쓴풀 발효물의 제조방법과 발효물을 함유한 상처치유 및 피부 재생, 진정, 항염 화장료 조성물에 관한 것이다.The present invention relates to a cosmetic composition, a fermented product for cosmetics, and a method for manufacturing the same, and in particular, Lactobacillus kurbatus J2K-01 is used as an inoculation strain, and the fermentation target is soothing the skin using centella, hollyhock, blue lotus, and bitter grass. Regarding the manufacturing method of fermented centella, hollyhock, blue lotus and bitter grass using Lactobacillus, which enhances wound healing and regeneration, and skin inflammation improvement effect, and wound healing and skin regeneration, soothing, anti-inflammatory cosmetic composition containing fermented products will be.
외부 환경을 접할 때 피부는 물리적, 화학적 자극 및 병원균으로부터 신체를 보호하는 매우 중요한 역할을 수행한다. When in contact with the external environment, the skin plays a very important role in protecting the body from physical and chemical irritation and pathogens.
피부 단면은 표피, 진피, 피하조직의 3개의 층으로 나누어지는데, 그 중에서도 표피의 각질층은 약 10~20%의 수분을 함유하고 인체의 최외각에 존재하여 체내로부터의 수분 증발을 억제하는 한편 외부로부터의 물질 과잉 침투를 차단한다. The cross section of the skin is divided into three layers: epidermis, dermis, and subcutaneous tissue. Among them, the stratum corneum of the epidermis contains about 10-20% of moisture and exists on the outermost part of the human body to inhibit water evaporation from the body. Blocks excessive penetration of material from
그러나 환경 변화, 생활습관, 사회 환경으로부터의 각종 스트레스와 공해물질로 인한 환경오염, 화장 습관에 따른 잦은 세안 및 노화에 따른 자연적인 피부 퇴화 등의 원인으로 피부가 건조해지고 표면이 거칠게 되며 피부가 윤기를 잃어 칙칙하게 보이는 등의 현상이 발생하게 되면서 피부건강에 관한 문제는 그 중요성이 점점 더 커지고 있다.However, due to environmental changes, lifestyle habits, environmental pollution due to various stresses and pollutants from the social environment, frequent cleansing according to makeup habits, and natural skin deterioration due to aging, the skin becomes dry, the surface becomes rough, and the skin becomes shiny. As phenomena such as loss of skin and dull appearance occur, problems related to skin health are becoming increasingly important.
피부 스트레스로부터 피부 건강을 유지하는 방법으로, 피부 수분의 건조 방지 및 각질층에서의 수분 보유능 향상을 위한 보습용 소재 개발에 많은 공을 들여 왔다. As a method of maintaining skin health from skin stress, much effort has been devoted to the development of moisturizing materials to prevent drying of skin moisture and improve moisture retention in the stratum corneum.
건조 피부가 형성되면 피부 표면의 방어 기능 (barrier function)이 소실되고 외부로부터의 자극물이나 알러젠의 피부 침입이 용이할 뿐만 아니라, 이러한 침입물에 대하여 피부가 거부 반응을 일으키기 때문이다.This is because when dry skin is formed, the barrier function of the skin surface is lost, and irritants or allergens from the outside can easily invade the skin, and the skin reacts to such intrusions.
피부의 거부반응은 표층 성분의 대다수를 구성하는 케라티노사이트 (keratinocyte)라는 각화 세포가 소수의 랑게르한스 (Langerhans), 멜라노사이트 (melanocyte) 등의 세포를 활성화시킴으로써, 이들 세포가 생산 및 방출하는 사이토카인 (cytokine)에 의한 염증 현상으로 나타난다. Rejection of the skin is a cytokine produced and released by keratinocytes, which constitute the majority of the surface layer, by activating a small number of cells such as Langerhans and melanocytes. It appears as an inflammatory phenomenon caused by (cytokine).
상기 사이토카인은 염증 세포를 불러모으는 작용을 하고, 특히 피부의 구성 세포를 자극시켜 피부 상층을 중심으로 한 염증 반응을 일으키고 습진 반응을 형성하는데, 이를 비알러지성 피부병이라고 한다.The cytokine functions to attract inflammatory cells, and in particular, by stimulating the constituent cells of the skin, it causes an inflammatory reaction centered on the upper layer of the skin and forms an eczema reaction, which is called a non-allergic skin disease.
따라서, 피부진정 및 피부보습 효과가 강력한 물질은 항염효과까지 확장될 수 있는 잠재적 가능성이 있고, 반대로 항염효과가 있는 물질이 피부진정 및 피부보습에도 효과가 있을 것으로 예측할 수 있으나 실질적으로 이와 같은 천연소재를 찾는 것은 매우 어려운 과정을 거쳐야 하는 문제가 있어 왔다.Therefore, substances with strong skin soothing and skin moisturizing effects have the potential to extend to anti-inflammatory effects, and on the contrary, substances with anti-inflammatory effects can be predicted to be effective in skin soothing and skin moisturizing. There has been a problem of having to go through a very difficult process to find.
이에 따른 필요에 의해 발견된 식물로 병풀이 있다.Centella asiatica is a plant that has been discovered by necessity.
병풀은 미나리과 식물로 호랑이풀이라고도 불리며, 오래전부터 항염, 해열, 항균 등에 효과가 있는 것으로 알려져 민간요법으로 널리 사용되어 왔다. Centella asiatica is a family of parsley, also called tiger grass, and has long been widely used as a folk remedy, known to have anti-inflammatory, antipyretic, and antibacterial effects.
구체적으로, 병풀의 잎과 줄기는 마데카식산(madecassic acid)이라는 성분을 갖는데, 이 성분은 지친 피부를 진정시키고 생기를 부여하며 피부 보호 및 흔적 관리에 도움을 주며, 손상 피부(염증)를 낫게 하고 상처 치료에도 효과적인 것으로 알려져 있다.Specifically, the leaves and stems of Centella asiatica have an ingredient called madecassic acid, which soothes and gives vitality to tired skin, helps protect skin and manages traces, and heals damaged skin (inflammation). It is known to be effective in treating wounds.
이에 "피부 트러블 예방 및 개선용 화장료 조성물"(한국 공개특허공보 제10-2019-0142007호, 특허문헌 1)에는 병풀 추출물 외에 어성초, 버드나무 가지, 캐모마일, 녹차 등의 추출물이 함께 포함되는 화장료 조성물 제시되어 있다.Accordingly, the "cosmetic composition for preventing and improving skin troubles" (Korean Patent Application Publication No. 10-2019-0142007, Patent Document 1) contains extracts such as Eoseongcho, willow branch, chamomile, green tea, etc. in addition to centella asiatica extract. Is presented.
촉규화는 아욱과의 접시꽃의 꽃을 말하는데, 꽃 모양이 접시를 닮아서 접시꽃이라고 부르며 백색 꽃을 백규화, 적색 꽃을 적규화라고 한다.Chok-gyu-hwa refers to the flower of a hollyhock of the mallow family. The flower shape resembles a plate, so it is called a hollyhock, white flowers are called white flowers, and red flowers are called red flowers.
촉규화는 종래에 백대하, 하복부 냉증, 대소변 불통, 토혈 등의 증상에 대해 약재로 사용하여 왔는데, "한약재 혼합 추출물을 함유하는 화장료 조성물"(한국 등록특허공보 제10-1135264호, 특허문헌 2)에는 자초(지치 뿌리), 구백(패랭이 꽃) 등과 조합하여 콜라겐 생합성 효과, 콜라게나아제 발현 억제, 엘라스타아제 발현 억제, 멜라닌 생성 억제 등에 효과를 갖는 화장료 조성물의 제조에 사용한 예가 공개되어 있다.Chok-gyu-hwa has been used as a medicinal material for symptoms such as baekdaeha, lower abdominal coldness, dyspnea, and hematopoiesis, but "cosmetic composition containing herbal medicinal mixture extract" (Korean Registered Patent Publication No. 10-1135264, Patent Document 2) , An example used in the manufacture of a cosmetic composition having effects on collagen biosynthesis, collagenase expression suppression, elastase expression suppression, melanin production suppression, etc. in combination with native grass (branch root), chinensis (dianthus) and the like is disclosed.
한편, 상기한 천연소재의 단순 추출물의 경우 추출 과정에서 물리 화학적으로 유효 성분이 변성되어 체내 흡수가 어려운 상태가 되거나 효과가 저감되는 문제점이 있다.On the other hand, in the case of the simple extract of the natural material described above, there is a problem in that the active ingredient is physically and chemically denatured during the extraction process, making it difficult to absorb in the body or reducing the effect.
이러한 문제를 해소하기 위해 최근에는 각종 균주를 이용하여 추출물을 발효하여 화장료 원료로 사용하는 기술들이 다수 공개되어 있다.In order to solve this problem, in recent years, a number of technologies for fermenting extracts using various strains and using them as raw materials for cosmetics have been disclosed.
일예로, "과채류 발효물 제조방법 및 과체료 발효물을 유효성분으로 함유하는 화장료 조성물"(한국 등록특허공보 제10-1905517호, 특허문헌 3)에는 락토바실러스 속 유산균을 과채류에 접종한 후 일정 조건에서 배양 및 발효시켜 발효물을 수득하고, 이를 화장료 원료로 사용하도록 한 바 있다.As an example, in "A method for manufacturing fermented fruit and vegetables and a cosmetic composition containing fermented fruit and fermented fruit as an active ingredient" (Korean Patent Publication No. 10-1905517, Patent Document 3), after inoculation of Lactobacillus sp. It has been cultivated and fermented under conditions to obtain a fermented product, which has been used as a raw material for cosmetics.
그러나, 특정 식물의 추출물, 이에 적용되는 특정 균주의 접종 및 발효를 통해 피부 진정 및 염증 개선에 대한 연구는 아직도 부족한 실정이며, 특히 쓴풀이나 푸른연꽃 등에 대한 연구는 더 부족한 실정이다.However, studies on skin soothing and inflammation improvement through inoculation and fermentation of specific plant extracts and specific strains applied thereto are still insufficient. In particular, studies on bitter grass or blue lotus are still insufficient.
본 발명은 상기와 같은 문제점을 해결하고자 연구 개발된 것으로 피부 진정, 염증 개선, 상처의 치유 재생에 효과가 있는 발효물의 제조 방법을 제공하려는 데 그 목적이 있다.The present invention has been researched and developed to solve the above problems, and an object of the present invention is to provide a method for producing a fermented product effective in soothing skin, improving inflammation, and healing wounds.
또한, 본 발명은 락토바실러스 쿠르바투스 J2K-01(기탁번호: KCTC 13494BP)로 발효한 발효물의 제조 방법을 제공함으로써 효과가 우수한 화장료 조성물을 제공하려는 것이다.In addition, the present invention is to provide a cosmetic composition excellent in effect by providing a method for producing a fermented product fermented with Lactobacillus curbatus J2K-01 (Accession No.: KCTC 13494BP).
본 발명의 락토바실러스 쿠르바투스 J2K- 01 균주와 쓴풀 등을 이용한 화장료용 발효물의 제조 방법은 상기한 과제를 해결하기 위하여, 병풀, 접시꽃, 푸른 연꽃, 쓴풀을 혼합 및 추출하여 혼합추출물을 수득하고, 상기 혼합추출물에 락토바실러스 쿠르바투스 J2K-01(기탁번호: KCTC 13494BP)을 접종하여 발효시킨 후, 침전물과 유산균체를 제거하여 발효물을 제조하는 것을 특징으로 한다.In order to solve the above problems, the manufacturing method of a fermented product for cosmetics using Lactobacillus kurbatus J2K-01 strain and bitter grass of the present invention is to obtain a mixed extract by mixing and extracting centella, hollyhock, blue lotus, and bitter grass. , After fermentation by inoculating Lactobacillus curbatus J2K-01 (accession number: KCTC 13494BP) to the mixed extract, the fermentation product is prepared by removing the precipitate and lactic acid bacteria.
상기한 구성에 있어서, 상기 혼합추출물 제조를 위한 원료는, 병풀 100 중량부를 기준으로, 접시꽃 50 ~ 150 중량부, 푸른 연꽃 50 ~ 150 중량부, 쓴풀 200 ~ 300 중량부가 혼합되어 구성된 것을 특징으로 한다.In the above configuration, the raw material for the preparation of the mixed extract is characterized in that, based on 100 parts by weight of centella asiatica, 50 to 150 parts by weight of hollyhock, 50 to 150 parts by weight of blue lotus, and 200 to 300 parts by weight of bitter grass are mixed. .
아울러, 본 발명의 발효물은 상기 제조 방법에 의해 제조된 것을 특징으로 한다.In addition, the fermented product of the present invention is characterized in that it is produced by the above manufacturing method.
본 발명의 락토바실러스 쿠르바투스 J2K- 01 균주와 쓴풀 등을 이용한 화장료 조성물은 상기 발효물이 화장료 조성물 전체 함량에 대해 0.001 ~ 10 중량% 함유되어 있는 것을 특징으로 한다.The cosmetic composition using the Lactobacillus curbatus J2K-01 strain and bitter grass of the present invention is characterized in that the fermentation product contains 0.001 to 10% by weight based on the total content of the cosmetic composition.
이때, 상기 화장료 조성물은 피부 염증 개선용인 것을 특징으로 한다.At this time, the cosmetic composition is characterized in that for improving skin inflammation.
또는, 상기 화장료 조성물은 피부 진정용인 것을 특징으로 한다.Or, the cosmetic composition is characterized in that for skin soothing.
또는, 상기 화장료 조성물은 상처 치유 및 재생용인 것을 특징으로 한다.Alternatively, the cosmetic composition is characterized in that for wound healing and regeneration.
본 발명에 의해, 피부 진정, 염증 개선, 상처 치유 및 재생에 효과가 있는 화장료 조성물이 제공된다.According to the present invention, there is provided a cosmetic composition effective in skin soothing, inflammation improvement, wound healing and regeneration.
도 1은 본 발명에서 발효물의 제조 방법을 나타낸 공정도.Figure 1 is a process diagram showing a method of manufacturing a fermented product in the present invention.
병풀은 미나리과 식물로 호랑이풀이라고도 불리며, 오래전부터 항염, 해열, 항균 등에 효과가 있는 것으로 알려져 민간요법으로 널리 사용되어 왔다. Centella asiatica is a family of parsley, also called tiger grass, and has long been widely used as a folk remedy, known to have anti-inflammatory, antipyretic, and antibacterial effects.
구체적으로, 병풀의 잎과 줄기는 마데카식산(madecassic acid)이라는 성분을 갖는데, 이 성분은 지친 피부를 진정시키고 생기를 부여하며 피부 보호 및 흔적 관리에 도움을 주며, 손상 피부(염증)를 낫게 하고 상처 치료에도 효과적인 것으로 알려져 있다.Specifically, the leaves and stems of Centella asiatica have an ingredient called madecassic acid, which soothes and gives vitality to tired skin, helps protect skin and manages traces, and heals damaged skin (inflammation). It is known to be effective in treating wounds.
접시꽃은 촉규화라고도 불리며 이는 접시꽃의 꽃을 말한다.The hollyhock is also called chokgyuhwa, and it refers to the flower of hollyhock.
접시꽃은 꽃 모양이 접시를 닮아서 접시꽃이라고 부르며 백색 꽃을 백규화, 적색 꽃을 적규화라고 한다.A hollyhock is called a hollyhock because its flower shape resembles a plate, and a white flower is called a white flower, and a red flower is called a red flower.
촉규화는 종래로부터 백대하, 하복부 냉증, 대소변 불통, 토혈 등의 증상에 대해 약재로 사용하여 왔다.[0003] Chok-gyuhwa has conventionally been used as a medicinal agent for symptoms such as baekdaeha, lower abdominal coldness, dyspepsia, and hematopoiesis.
푸른 연꽃(blue lotus)는 남아프리카가 원산지인 여러해살이풀로서, 태국산 수련(Nymphaea coerulea), 인도산 수련(Nymphaea, stellata)등도 이에 속한다. Blue lotus is a perennial plant native to South Africa, and includes Thai water lily (Nymphaea coerulea) and Indian water lily (Nymphaea, stellata).
수련과 식물은 지름 30cm 정도의 잎을 가진 매우 크게 자라는 야생수초로서 남부아시아와 동아시아 인근의 섬들에서 자란다. 보통 수련이라고 불리는 이 식물은 푸른색 꽃이 피는 데서 블루로터스(blue lotus)라고 불린다The water lily family is a wild aquatic plant that grows very large with leaves of about 30cm in diameter, and grows on islands near southern and eastern Asia. This plant, usually called water lily, is called blue lotus because it has blue flowers.
쓴풀은 당약(Swertia japonica MAKINO) 또는 고초라고도 불리는 한해살이 또는 두해살이 풀로, 뿌리에는 쓴맛이 강한 성분을 함유하고 있다. Bitter grass is a herbaceous herb called Swertia japonica MAKINO or gocho. It contains a strong bitter taste in its roots.
모가 져 있는 줄기는 위로 곧게 서서 위쪽 부분에서 가지를 친다. 줄기는 자줏빛이 돌며 높이는 15~30cm 가량이다. The stalk with hair is standing upright and pruned at the upper part. The stem is purple and the height is about 15~30cm.
잎은 피침 꼴이고 잎자루를 가지지 않으며 마디마다 2장이 마주 난다. 꽃은 줄기 끝에 원뿌리 꼴로 모여 피어나는데 위에서부터 아래로 차례로 피어 나가는 습성을 가지고 있다. The leaves are lanceolate, do not have petioles, and two are facing each other. Flowers gather in the shape of a root at the end of the stem and bloom in order from top to bottom.
길쭉한 타원 꼴인 5장의 꽃잎을 가지고 있으며 꽃잎은 피어남에 따라 완전히 펼쳐진다. 꽃의 지름은 1.5~2cm이며 흰 바탕에 자주색 줄이 나 있기 때문에 연보랏빛으로 보인다. 꽃의 한가운데에는 5개의 수술이 자리하고 있는데 꽃가루주머니는 짙은 보랏빛이다. It has 5 petals of an elongated oval shape, and the petals are fully spread out as they bloom. The diameter of the flower is 1.5~2cm, and it looks light purple because there are purple lines on the white background. Five stamens are located in the middle of the flower, and the pollen pouch is dark purple.
분포는 제주도와 남부 지방에 주로 분포하며 산과 들판의 양지바른 풀밭에 난다. Distribution is mainly distributed in Jeju Island and southern regions, and it grows in sunny meadows in mountains and fields.
민간 의학 분야에서는 소화 불량과 식욕부진, 구충 약 등으로 사용되어 왔다.In the field of private medicine, it has been used as a medicine for indigestion, anorexia, and anthelmintic.
최근에 모발 화장품 분야에서 사용되는 보고가 있으나, 피부 진정 및 염증 완화에 관해 사용된 바는 알려진 바가 없다.Recently, there have been reports of being used in the field of hair cosmetics, but there is no known one used for soothing skin and alleviating inflammation.
락토바실러스 쿠르바투스 J2K-01(기탁번호: KCTC 13494BP)은 최근 "피부 탄력 및 주름개선용 화장료 조성물"(한국 등록특허공보 제10-1930342호)에 공개된 신규 락토바실러스 쿠르바투스 균주로, 염기서열 및 기탁증은 상기 특허 공보에 공개되어 있다.Lactobacillus Courbatus J2K-01 (Accession No.: KCTC 13494BP) is a new Lactobacillus Courbatus strain recently published in “a cosmetic composition for improving skin elasticity and wrinkles” (Korean Patent Publication No. 10-1930342), The nucleotide sequence and the deposit certificate are disclosed in the above patent publication.
병풀, 접시꽃, 푸른 연꽃, 쓴풀은 필요에 따라 잎, 줄기, 꽃, 뿌리 등을 전체 또는 부분적으로 추출하여 사용할 수 있다.Centella asiatica, hollyhock, blue lotus, and bitter grass can be used by extracting all or part of leaves, stems, flowers, and roots as needed.
보다 바람직하기로는 병풀과 쓴풀은 전초를 사용하고, 접시꽃과 푸른연꽃은 꽃 부분을 활용함이 바람직하다.More preferably, centella and bitter plants use outposts, and hollyhock and blue lotus flowers are preferably used.
본 발명의 락토바실러스 쿠르바투스 J2K- 01 균주와 쓴풀 등을 이용한 화장료용 발효물의 제조 방법은 도 1에 나타나 있는 바와 같이 원료를 혼합추출한 다음, 멸균시키고, 이어 미생물을 이용하여 배양 발효시킨 다음, 원심분리 및 감압 농축 단계를 거쳐 진공 건조함으로써 제조된다. The manufacturing method of a cosmetic fermentation product using the Lactobacillus kurbatus J2K-01 strain and bitter grass of the present invention, as shown in Fig. 1, mixed and extracted raw materials, sterilized, and then cultured and fermented using microorganisms, It is produced by vacuum drying through centrifugation and vacuum concentration steps.
보다 구체적으로 원료로 병풀, 접시꽃, 푸른 연꽃, 쓴풀을 사용하며, 이들을 개별 추출 후 추출물을 혼합하거나, 혼합한 다음 추출하여 혼합추출물을 수득할 수 있다.More specifically, centella asiatica, hollyhock, blue lotus, and bitter grass are used as raw materials, and extracts may be mixed after individual extraction, or mixed and then extracted to obtain a mixed extract.
이때, 바람직하기로는 원료의 혼합 후 추출함이 바람직하다.At this time, it is preferable to extract the raw materials after mixing.
원료의 함량비는 병풀 0.1 ~ 30 질량비, 접시꽃 0.1 ~ 30 질량비, 푸른연꽃 0.1 ~ 30 질량비, 쓴풀 1 ~ 50 질량비로 구성될 수 있다.The content ratio of the raw material may consist of 0.1 to 30 mass ratio of centella, 0.1 to 30 mass ratio of hollyhock, 0.1 to 30 mass ratio of blue lotus, and 1 to 50 mass ratio of bitter grass.
보다 구체적으로는 병풀 100 중량부를 기준으로, 접시꽃 50 ~ 150 중량부, 푸른연꽃 50 ~ 150 중량부, 쓴풀 200 ~ 300 중량부가 혼합되어 구성될 수 있다.More specifically, based on 100 parts by weight of centella asiatica, 50 to 150 parts by weight of hollyhock, 50 to 150 parts by weight of blue lotus, and 200 to 300 parts by weight of bitter grass may be mixed.
추출 방식은 열수 추출, 증류 추출, 에탄올 추출, 초음파 추출 등 다양한 방식이 적용 가능하나, 80 ~ 100℃의 정제수를 사용하여 1 ~ 6시간 추출하는 것이 적절하다.As for the extraction method, various methods such as hot water extraction, distillation extraction, ethanol extraction, ultrasonic extraction, etc. can be applied, but it is appropriate to extract for 1 to 6 hours using purified water at 80 ~ 100℃.
보다 바람직하기로는 85~95℃의 온도로 3 ~ 5시간 동안 추출하는 것이 좋다.More preferably, it is good to extract for 3 to 5 hours at a temperature of 85 to 95 °C.
더불어, 추출에 의해 혼합추출물이 제조되면, 이를 냉각 및 여과시키고, 120℃ 내외의 온도에서 10 ~ 20분간 멸균 처리한다.In addition, when the mixed extract is prepared by extraction, it is cooled and filtered, and sterilized for 10 to 20 minutes at a temperature of about 120°C.
혼합추출물의 준비와 더불어, 기탁번호 KCTC 13494BP의 락토바실러스 쿠르바투스 J2K-01을 접종하여 배양 및 발효시킨다.In addition to the preparation of the mixed extract, Lactobacillus curbatus J2K-01 of accession number KCTC 13494BP was inoculated, cultured and fermented.
배양 및 발효 조건은 온도 25 ~ 32℃가 적정하고, 배양 기간은 3일 내지 5일 정도가 적절하다.The culture and fermentation conditions are appropriate at a temperature of 25 to 32°C, and a culture period is appropriate for about 3 to 5 days.
배양 및 발효에 의해 발효액이 수득되면 이를 원심분리기에 투입하여 8,000 ~ 1,2000 rpm의 조건으로 15 ~ 25분 동안 원심분리하여 침전물과 유산균체가 제거된 발효물을 수득한다.When the fermentation broth is obtained by cultivation and fermentation, it is added to a centrifuge and centrifuged for 15 to 25 minutes under the conditions of 8,000 to 1,2000 rpm to obtain a fermented product from which precipitates and lactic acid bacteria are removed.
그런 다음 발효물을 감압 농축 및 진공 건조 처리해준다.Then, the fermented product is concentrated under reduced pressure and vacuum-dried.
한편, 상기와 같이 제조된 발효물은 화장료 조성물 원료로 포함되는데, 적절한 함량은 화장료 조성물 전체 함량에 대해 0.001 ~ 10 중량% 함유되는 것이 좋다.Meanwhile, the fermented product prepared as described above is included as a raw material for the cosmetic composition, and an appropriate content is preferably contained in 0.001 to 10% by weight based on the total content of the cosmetic composition.
이하, 실시예를 통해 본 발명을 좀 더 구체적으로 설명한다. 단, 이들 실시예는 본 발명의 예시적인 기재일 뿐이며, 본 발명의 범위가 이들 실시예에 국한되는 것은 아니다.Hereinafter, the present invention will be described in more detail through examples. However, these examples are only illustrative descriptions of the present invention, and the scope of the present invention is not limited to these examples.
실시예 1: 발효물의 제조Example 1: Preparation of fermented product
병풀 전초 20g, 접씨꽃의 꽃 20g, 푸른연꽃의 꽃 20g, 쓴풀 전초 50g을 추출 용기에 넣고 정제수 1000g을 혼합한 다음 90℃에서 3시간 동안 추출하여 혼합추출물을 제조하고 냉각하여 여과하였다. 20g Centella asiatica, 20g flowers, 20g blue lotus flowers, and 50g bitter grass were placed in an extraction container, mixed with 1000g of purified water, and extracted for 3 hours at 90°C to prepare a mixed extract, cooled, and filtered.
그런 다음 배양을 위해 혼합추출물을 121℃에서 15분간 멸균하였다. Then, the mixed extract was sterilized at 121°C for 15 minutes for culture.
멸균이 끝난 혼합추출물에 락토바실러스 쿠르바투스 J2K-01을 접종하여 온도 32℃, 정치배양 조건으로 3일간 배양 발효시켜 발효액을 제조하였다.The sterilized mixed extract was inoculated with Lactobacillus kurbatus J2K-01 and cultured and fermented for 3 days at a temperature of 32° C. and stationary culture conditions to prepare a fermentation broth.
이어 상기 발효액을 10,000 rpm에서 20분간 원심분리하여 침전물과 유산균체가 제거된 발효물을 얻었다. Subsequently, the fermentation broth was centrifuged at 10,000 rpm for 20 minutes to obtain a fermented product from which precipitates and lactic acid bacteria were removed.
이를 감압농축 및 진공건조하여 분말을 얻었다.This was concentrated under reduced pressure and dried under vacuum to obtain a powder.
실시예 2 : 발효물의 제조Example 2: Preparation of fermented product
실시예 1과 동일하게 진행하되, 접시꽃은 9g을 사용하였다.Proceed in the same manner as in Example 1, but 9g of hollyhock was used.
실시예 3 : 발효물의 제조Example 3: Preparation of fermented product
실시예 1과 동일하게 진행하되, 접시꽃은 31g을 사용하였다.Proceed in the same manner as in Example 1, but 31g of hollyhock was used.
실시예 4 : 발효물의 제조Example 4: Preparation of fermented product
실시예 1과 동일하게 진행하되, 푸른연꽃은 9g을 사용하였다.Proceed in the same manner as in Example 1, but 9g of blue lotus was used.
실시예 5 : 발효물의 제조Example 5: Preparation of fermented product
실시예 1과 동일하게 진행하되, 푸른연꽃은 31g을 사용하였다.Proceed in the same manner as in Example 1, but 31g of blue lotus was used.
실시예 6 : 발효물의 제조Example 6: Preparation of fermented product
실시예 1과 동일하게 진행하되, 쓴풀은 39g을 사용하였다.Proceed in the same manner as in Example 1, but 39g of bitter grass was used.
실시예 7 : 발효물의 제조Example 7: Preparation of fermented product
실시예 1과 동일하게 진행하되, 쓴풀은 61g을 사용하였다.Proceed in the same manner as in Example 1, but 61g of bitter grass was used.
비교예 1: 혼합 추출물의 제조Comparative Example 1: Preparation of mixed extract
병풀20g, 접씨꽃 20g, 푸른연꽃 20g, 쓴풀 50g을 추출 용기에 넣고 정제수 1000g을 혼합한 추출물을 90℃에서 3시간 추출하고 냉각하여 여과하였다. 상기 추출물을 감압농축 및 진공건조하여 분말을 얻었다. Centella asiatica 20g, Phalaenopsis 20g, blue lotus 20g, and bitter grass 50g were placed in an extraction container, and the extract mixed with 1000g of purified water was extracted for 3 hours at 90°C, cooled, and filtered. The extract was concentrated under reduced pressure and dried under vacuum to obtain a powder.
비교예 2 : 발효물의 제조Comparative Example 2: Preparation of fermented product
실시예 1과 동일하게 진행하되, 균주로 락토바실러스 펜토서스 J2K-185를 접종하였다.Proceed in the same manner as in Example 1, but inoculated with Lactobacillus pentosus J2K-185 as a strain.
비교예 3 : 발효물의 제조Comparative Example 3: Preparation of fermented product
실시예 1과 동일하게 진행하되, 균주로 락토바실러스 브레비스 J2K-55를 접종하였다.Proceed in the same manner as in Example 1, but inoculated with Lactobacillus brevis J2K-55 as a strain.
비교예 4 : 발효물의 제조Comparative Example 4: Preparation of fermented product
실시예 1과 동일하게 진행하되, 균주로 락토바실러스 쿠르바투스 AMI-1214를 접종하였다.Proceed in the same manner as in Example 1, but inoculated with Lactobacillus curbatus AMI-1214 as a strain.
비교예 5 : 발효물의 제조Comparative Example 5: Preparation of fermented product
실시예 1과 동일하게 진행하되 원료 중 병풀을 배제한 체 진행하였다.Proceeding in the same manner as in Example 1, except for excluding centella from the raw materials.
비교예 6 : 발효물의 제조Comparative Example 6: Preparation of fermented product
실시예 1과 동일하게 진행하되, 원료 중 접시꽃을 배제한 채 진행하였다.Proceed in the same manner as in Example 1, except for the hollyhock among the raw materials.
비교예 7 : 발효물의 제조Comparative Example 7: Preparation of fermented product
실시예 1과 동일하게 진행하되, 원료 중 푸른 연꽃을 배제한 채 진행하였다.Proceed in the same manner as in Example 1, except for the blue lotus flower among the raw materials.
비교예 8 : 발효물의 제조Comparative Example 8: Preparation of fermented product
실시예 1과 동일하게 진행하되, 원료 중 쓴풀을 배제한 채 진행하였다.Proceed in the same manner as in Example 1, except for bitter grass among the raw materials.
<실험예 1> 피부 개선 효과 평가<Experimental Example 1> Evaluation of skin improvement effect
실시예들 및 비교예들의 추출물을 이용하여 아래 표 1에 조성의 크림 형태의 화장료를 제조하였다.Using the extracts of Examples and Comparative Examples, a cosmetic composition in the form of a cream having the composition shown in Table 1 below was prepared.
표에서 상기 실시예 1 내지 7, 비교예 1 내지 8에 의해 제조된 추출물은 화장료 전체 조성의 1중량%가 되도록 조정하였다.In the table, the extracts prepared according to Examples 1 to 7, Comparative Examples 1 to 8 were adjusted to be 1% by weight of the total composition of the cosmetic.
그런 다음 실시예 및 비교예의 추출물을 사용한 경우의 피부 재생 및 상처의 치유 개선 정도를 비롯하여 피부 보습, 미백 효과, 피부 진정 효과를 평가하였다.Then, when the extracts of Examples and Comparative Examples were used, the degree of skin regeneration and wound healing improvement, skin moisturizing, whitening effect, and skin soothing effect were evaluated.
구체적으로 레이저 시술 또는 박피 시술을 받은 대상자를 대상으로 실시예 및 비교예의 화장료를 각각 5명씩 일주일 동안(1일 2번(아침, 저녁으로)) 사용하도록 한 후, 10점 척도법(1: 대단히 나쁘다 --> 10:대단히 좋다)으로 평가한 다음 Duncan's multiple range test로 유의성을 검정하였다. Specifically, for subjects who have undergone laser treatment or dermabrasion treatment, use the cosmetics of Examples and Comparative Examples for 5 people each for a week (twice a day (in the morning and evening)), and then the 10-point scale method (1: very bad) --> 10: Very good), and then the significance was tested with Duncan's multiple range test.
그 결과 하기 표 2와 같은 성적을 얻었다.As a result, the results shown in Table 2 were obtained.
(항염 효과)Wound healing
(Anti-inflammatory effect)
상기 표 2에서 보듯이 실시예들은 비교예들에 비해 전반적으로 피부 재생, 상처 치유 효과가 뛰어난 것으로 나타났으며, 피부 보습, 미백, 피부 진정 효과를 비롯하여 전체적인 선호도가 더 높은 것으로 나타났다.As shown in Table 2 above, the Examples showed superior overall skin regeneration and wound healing effects compared to the Comparative Examples, and overall preference was found to be higher, including skin moisturizing, whitening, and skin soothing effects.
<실험예 2> 피부 자극 시험<Experimental Example 2> Skin irritation test
상기 실험예 1과 같이 크림형의 화장료를 제조한 후, 각 화장료에 대한 피부 자극 시험을 조사하기 위해 피부 첩포시험을 실시하였다.After preparing a cream-shaped cosmetic as in Experimental Example 1, a skin patch test was performed to investigate the skin irritation test for each cosmetic.
피검자는 15~30세의 시험 부위에 피부 질환이 없는 사람으로 선정하였으며 총 15명에게 실시하였다. 첩포부위인 상박부를 70% 에탄올로 닦아내고 건조시킨 후, 제품을 01g씩 가하여 인체 상박부에 24시간 첩포하고, 첩포 제거 후 1시간 이내에 피부 반응을 검사하고, 다음날(48시간 후) 다시 검사하였다. 피부 반응 판정은 하기 검사기준에 의해 판정하였다. 피부 첩포 시험 결과를 표 4에 나타내었다.Subjects were selected as those without skin disease at the test site of 15-30 years old, and a total of 15 subjects were conducted. After wiping and drying the upper arm, which is the patch area, with 70% ethanol, 01 g of the product was added to the human upper arm for 24 hours, and the skin reaction was examined within 1 hour after removal of the patch, and the next day (after 48 hours) was examined again. The skin reaction was determined according to the following test criteria. Table 4 shows the results of the skin patch test.
* 피부 반응 검사기준* Skin reaction test criteria
- 반응 없음- no response
± 미약한 양성 반응 (홍반)± weak positive reaction (erythema)
+ 양성 반응 (홍반)+ Positive reaction (erythema)
++ 강한 양성 반응 (홍반, 부종)++ strong positive reaction (erythema, edema)
+++ 심한 양성 반응 (홍반, 부종, 수포)+++ Severe positive reaction (erythema, swelling, blisters)
상기 표 3에서 보듯이 실시예의 경우 피부 자극이 거의 없는 것으로 나타났다.As shown in Table 3, in the case of the Example, it was found that there was little skin irritation.
<실험예 3> 염증유발관련 효소 (hyaluronidase) 활성 억제효과 측정<Experimental Example 3> Measurement of the inhibitory effect of hyaluronidase activity
실시예 및 비교예의 추출물의 항염증 효과를 측정하기 위하여 염증유발효소인 히아루로니디아제 (hyaluronidase) 활성 억제효과를 측정하였다. 히아루로니디아제 (hyaluronidase)는 히아루론산을 가수분해하는 효소로 염증 유발하는 효소이다.In order to measure the anti-inflammatory effect of the extracts of Examples and Comparative Examples, the inhibitory effect of hyaluronidase activity, an inflammatory enzyme, was measured. Hyaluronidase is an enzyme that hydrolyzes hyaluronic acid and causes inflammation.
본 실험예에서는 히아루로니디아제 (hyaluronidase)의 활성을 억제하여 항염증효과를 측정하는 방법 (Kakegawa Y: Japanese J. of Inflammation, 4, 437-438, 1984)을 응용해 항염증 효과를 판정하였다. In this experimental example, the anti-inflammatory effect was determined by applying a method of measuring the anti-inflammatory effect by inhibiting the activity of hyaluronidase (Kakegawa Y: Japanese J. of Inflammation, 4, 437-438, 1984). .
시료 100㎕와 히아루로니디아제 용액 (type Ⅳ-S, Sigma, 400U/㎖) 50㎕를 넣고 37℃에서 20분간 반응시킨 후, 효소활성화용액 (compound 48/80 CaCl2ㆍ2H2O, Sigma, 0.1㎎/㎖)을 100㎕를 첨가하고 다시 37℃에서 20분간 반응시켰다. 100 µl of sample and 50 µl of hyaluronidiase solution (type IV-S, Sigma, 400U/ml) were added and reacted at 37°C for 20 minutes, followed by enzyme activation solution (compound 48/80 CaCl 2 ㆍ2H 2 O, Sigma , 0.1 mg/ml) was added and reacted again at 37°C for 20 minutes.
히아루론산 (hyaluronic acid) 용액(0.4㎎/㎖)을 250㎕ 넣고 37℃에서 40분간 반응시키고, 0.4N NaOH 100㎕를 넣어 반응을 종결시켰다. 포타슘보레이트 용액을 100㎕ 첨가하여 95℃에서 3분간 반응시키고 냉각시킨 다음 ρ-디메틸아미노벤즈알데히드 용액을 3㎖ 넣고 다시 20분간 37℃에서 반응시킨 후 발색시켰다. 585㎚에서 흡광도를 측정하여 히아루로니디아제 활성 저해율을 측정하였다.250 µl of hyaluronic acid solution (0.4 mg/ml) was added and reacted at 37° C. for 40 minutes, and 100 µl of 0.4N NaOH was added to terminate the reaction. 100 µl of potassium borate solution was added, reacted at 95° C. for 3 minutes, cooled, 3 ml of ρ-dimethylaminobenzaldehyde solution was added, and reacted at 37° C. for 20 minutes, followed by color development. The absorbance was measured at 585 nm to measure the hyaluronidase activity inhibition rate.
히아루로니디아제의 활성 저해율(%)은 수학식 1에 의하여 계산하였으며, IC50 값은 히아루로니디아제 효소 활성을 50% 저해하는 물질의 농도이다.The hyaluronidase activity inhibition rate (%) was calculated by Equation 1, and the IC 50 value is the concentration of a substance that inhibits the hyaluronidiase enzyme activity by 50%.
<수학식 1><Equation 1>
히아로로니다아제의 활성 저해율(%) = [(A-B)/A] ×100Hyaluronidase activity inhibition rate (%) = [(A-B)/A] × 100
A: 시료(실시예 및 비교예의 추출물)를 첨가하지 않은 웰의 효소활성A: Enzymatic activity of wells without addition of samples (extracts of Examples and Comparative Examples)
B: 시료(실시예 및 비교예의 추출물)를 첨가한 웰의 효소활성B: Enzymatic activity of wells to which samples (extracts of Examples and Comparative Examples) were added
히아루로니디아제 (hyaluronidase)의 활성 저해효과를 측정한 결과(표 4), 실시예 1의 혼합추출물의 IC50값이 0.36%로 나타나 비교예들에 비해 우수한 항염증 효과를 확인할 수 있었다.As a result of measuring the activity inhibitory effect of hyaluronidase (Table 4), the IC 50 value of the mixed extract of Example 1 was 0.36%, indicating superior anti-inflammatory effect compared to the comparative examples.
Claims (7)
병풀, 접시꽃, 푸른 연꽃, 쓴풀을 혼합 및 추출하여 혼합추출물을 수득하고,
상기 혼합추출물에 락토바실러스 쿠르바투스 J2K-01(기탁번호: KCTC 13494BP)을 접종하여 발효시킨 후, 침전물과 유산균체를 제거하여 발효물을 제조하는 것을 특징으로 하는,
락토바실러스를 이용한 병풀, 접시꽃, 푸른연꽃, 쓴풀 화장료용 발효물의 제조방법.
In the manufacturing method of fermented product for cosmetic,
Centella asiatica, hollyhock, blue lotus, and bitter grass were mixed and extracted to obtain a mixed extract,
After fermentation by inoculating Lactobacillus curbatus J2K-01 (accession number: KCTC 13494BP) to the mixed extract, the fermentation product is prepared by removing the precipitate and lactic acid bacteria,
A method for producing fermented products for cosmetic products of centella, hollyhock, blue lotus and bitter grass using lactobacillus.
상기 혼합추출물 제조를 위한 원료는,
병풀 100 중량부를 기준으로, 접시꽃 50 ~ 150 중량부, 푸른 연꽃 50 ~ 150 중량부, 쓴풀 200 ~ 300 중량부가 혼합되어 구성된 것을 특징으로 하는,
락토바실러스를 이용한 병풀, 접시꽃, 푸른연꽃, 쓴풀 화장료용 발효물의 제조방법.
The method of claim 1,
The raw material for preparing the mixed extract,
Based on 100 parts by weight of Centella asiatica, 50 to 150 parts by weight of hollyhock, 50 to 150 parts by weight of blue lotus, and 200 to 300 parts by weight of bitter grass are mixed.
A method for producing fermented products for cosmetic products of centella, hollyhock, blue lotus and bitter grass using lactobacillus.
제 1항 내지 제 2항 중 어느 한 항의 제조 방법에 의해 제조된 것을 특징으로 하는,
락토바실러스를 이용한 병풀, 접시꽃, 푸른연꽃, 쓴풀 화장료용 발효물.
In the cosmetic fermented product,
It characterized in that it is produced by the manufacturing method of any one of claims 1 to 2,
Fermented product for cosmetic products of centella, hollyhock, blue lotus, and bitter grass using lactobacillus.
제 3항의 화장료용 발효물이 화장료 조성물 전체 함량에 대해 0.001 ~ 10 중량% 함유되어 있는 것을 특징으로 하는,
락토바실러스를 이용한 병풀, 접시꽃, 푸른연꽃, 쓴풀 화장료용 발효물을 함유한 상처치유, 피부 재생, 진정, 및 항염 효능을 갖는 화장료 조성물.
In the cosmetic composition,
The cosmetic fermented product of claim 3 is characterized in that it contains 0.001 to 10% by weight based on the total content of the cosmetic composition,
A cosmetic composition having a wound healing, skin regeneration, soothing, and anti-inflammatory effect containing fermented products for centella, hollyhock, blue lotus, bitter herb cosmetics using Lactobacillus.
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