KR19990081226A - Skin cosmetic using natural mushrooms - Google Patents

Skin cosmetic using natural mushrooms Download PDF

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KR19990081226A
KR19990081226A KR1019980015036A KR19980015036A KR19990081226A KR 19990081226 A KR19990081226 A KR 19990081226A KR 1019980015036 A KR1019980015036 A KR 1019980015036A KR 19980015036 A KR19980015036 A KR 19980015036A KR 19990081226 A KR19990081226 A KR 19990081226A
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skin
extract
natural
situation mushroom
effect
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KR1019980015036A
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Korean (ko)
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김상묵
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이세복
나드리화장품주식회사
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Abstract

본 발명은 자연산 상황버섯을 이용한 피부화장료에 관한 것으르 피부개선효과 특히 피부미백효과와 항산화효과가 탁월하고 안전성이 우수한 자연산 상황버섯의 열수 추출물, 20-40% 에탄올 수용액 추출물,1,3-부틸렌글리콜, 프로필렌글리콜 및 물의 혼합액으로 추출한 자연산 상황버섯 추출물로서 피부화장료의 성분으로서 적용가능한 것이다.The present invention relates to a skin cosmetic using natural situation mushroom, hot water extract of natural situation mushroom, 20-40% ethanol aqueous extract, 1,3-butyl skin improvement effect, particularly skin whitening and antioxidant effect, excellent safety Natural situation mushroom extract extracted with a mixture of len glycol, propylene glycol and water is applicable as a component of skin cosmetics.

Description

자연산 상황버섯을 이용한 피부화장료Skin cosmetic using natural mushrooms

본 발명은 자연산 상황버섯을 이용한 피부화장료에 관한 것으로 특히 피부 미백효과, 항산화효과 등 피부개선효과가 탁월하여 피부화장료의 성분으로서 자연산 상황버섯의 열수추출물, 에탄올 추출물, 1,3-부틸렌글리콜, 프로필렌글리콜과 물의 혼합액으로 추출한 추출물을 함유한 피부화장료에 관한 것이다.The present invention relates to skin cosmetics using natural situation mushrooms, in particular, the skin improvement effect such as skin whitening effect, antioxidant effect, as a component of skin cosmetics, hot water extract of natural situation mushrooms, ethanol extract, 1,3-butylene glycol, It relates to a skin cosmetic containing an extract extracted with a mixture of propylene glycol and water.

일반 피부외용제에서는 화장품, 의약품, 의약부외품 등이 있고 특히 피부손상의 방지 및 개선을 목적으로 한 피부외용제에 대해서는 종래부터 천연물로부터 추출한 원료를 시작으로 여러가지의 원재료를 배합하여 사용하고 있다. 예를들면, 인삼추출물과 감초추출물, 송이버섯 추출물 등의 추출물, 콜라겐 등의 천연고분자,아미노산, 비타민 등의 피부세포활성제 혹은 글리세린, 1,3-부틸렌글리콜 등의 보습제 등이 피부손상의 방지 및 개선을 목적으로 피부화장료에 이용되어 오고있었다. 그러나 이들 종래의 피부손상 방지 및 개선에 이용되어 온 물질을 사용함에 있어 자연상태에서 채취는 물론 인공재배한 것을 사용하고있는데 같은 물질이라도 채취시기나 채취장소에 따라 효능효과가 다른 것은 공지의 사실이다(Proc,Jpn, Assoc, Mycotoxical, No.30,1989). 특히 식물의 경우에 있어 피부의 미백효과나 면역부활효과, 항산화효과같은 특이한 효과를 얻기위해서는 그 식물의 채취부위나 채취시기, 채취장소에 따라 효과가 많이 차이나는데 문제점이 있었다.In general external skin preparations, there are cosmetics, medicines, quasi-drugs, etc. In particular, external skin preparations for the purpose of preventing and improving skin damage have conventionally used various raw materials starting from raw materials extracted from natural products. For example, ginseng extract, licorice extract, extracts such as pine mushroom extract, natural polymers such as collagen, skin cell activators such as amino acids and vitamins, or moisturizing agents such as glycerin and 1,3-butylene glycol are used to prevent skin damage. And it has been used in skin cosmetics for the purpose of improvement. However, in the use of materials that have been used for preventing and improving such conventional skin damage, it is collected in nature and artificially cultivated. It is well known that the same substance has different efficacy effects depending on the time of collection and the place of collection. (Proc, Jpn, Assoc, Mycotoxical, No. 30,1989). In particular, in the case of plants, in order to obtain specific effects such as skin whitening effect, immune resiliency effect, and antioxidant effect, there was a problem that the effects vary greatly depending on the harvesting site, collection time, and collection location of the plant.

상황버섯[phellinus linteus (Ikekawa, T., : Antitumor action of some basidiemycetes, especially Phellinus Linteus. GANN,59 : 155,l968)]은 옛날부터 중국에서는 뽕나무에 기생하는 것으로 소개되어 있으나 우리나라에서 조사된바에 의하면 활엽수에서도 야생되는 것을 볼수 있다(차동렬, 월간잠사 p34-37,95.12). 우리나라에서 상황버섯이라하는 것은 목질진흙버섯(phellinus linteus)을 지칭하며 소나무 비늘버섯과(Hymenochaetaceae)에 속한다. 이 상황버섯속에 속하는 진흙버섯 속에는 전 세계적으로 약 48종이 보고되고 있으나 우리나라에서는 녹슨진흙버섯(P. ferruginosus), 마른진흙버섯(P. gilvus), 말똥진흙버섯(P. igniarius), 가지진흙버섯(P.laevigatus), 벚나무진흙버섯(P. pomaceus), 찰진흙버섯(P. robustus), 전나무진흙버섯, 낙엽진흙버섯(P. pini) 등으로 8종이 채집되었다.이러한 진흙버섯에 속하는 버섯들은 옛날부터 상이(桑耳), 침층공균(針層孔菌),호손안(胡孫眼) 등의 명칭으로 자궁출혈, 월경불순 등 부인병의 치료제로 이용되어 왔으며 현재까지 여러 연구자에 의해 밝혀진 바에 의하면 동물실험에서 종양저지율이 60-97%로 우수한 효과가 있는 것으로 알려지고 있다.Phellinus linteus (Ikekawa, T.,: Antitumor action of some basidiemycetes, especially Phellinus Linteus. GANN, 59: 155, l968) has been introduced since ancient times as a parasitic to mulberry trees in China. It is also found to be wild in hardwoods (Cha Dong-ryul, Monthly subp. P34-37,95.12). In Korea, the situation mushroom refers to the woody mud mushroom (phellinus linteus) and belongs to the pine scale Hymenochaetaceae. About 48 species of mud mushrooms belonging to this situation are reported around the world, but in Korea, rust mud mushrooms (P. ferruginosus), dried mud mushrooms (P. gilvus), worm mud mushrooms (P. igniarius), and branch mud mushrooms ( Eight species were collected from P. laevigatus, P. pomaceus, P. robustus, fir mud, and deciduous mushroom (P. pini). It has been used for the treatment of gynecological diseases such as uterine bleeding and menstrual irregularities under the names of differentiation, cotyledon, and Hawthorne. In the experiment, tumor suppression rate is 60-97% and is known to have an excellent effect.

이와같이 자연계에 자생하는 상황이 항종양성을 갖는 것은 공지의 사실이지만, 자연계에서는 번식하기 어렵고 실제 상황이라 부르는 뽕나무의 고목에 자생하는 상황은 뽕나무자체의 감소와 함께 구하기가 힘들게되어 현재는 인공재배법으로 국내에서도 재배하고 있다. 오래전부터 뽕나무의 줄기, 잎, 뿌리 등은 의약과 화장품 분야에서 이용되어져 오고 있으며 최근에는 인공재배한 상황버섯을 가지고 항암효과의 규명에 대한 연구만이 국내외에서 진행되고 있는 실정이다.As such, it is well known that a situation that grows naturally in nature has anti-tumor, but it is difficult to breed in nature and grows in an old tree of mulberry, which is called a real situation, and it is difficult to obtain with the decrease of mulberry itself. I grow it. The stems, leaves, and roots of mulberry trees have been used in medicine and cosmetics fields for a long time. Recently, only studies on the identification of anticancer effects with artificially cultivated situation mushrooms have been conducted at home and abroad.

본 발명은 앞에서 말한 종래기술의 문제점을 거울삼아 각종 피부손상의 방지 및 개선효과가 우수하고 안전성면에서도 우수한 피부화장료를 제공하는 것이다. 상기한 목적을 달성하기위한 본 발명의 자연산 상황버섯 추출물은 한반도의 북한지방의 자연상태에서 성장해온 상황버섯(P. pini와 P. linteus))을 가지고 열수추출물, 20-30% 에탄올추출물, 1,3-부틸렌글리콜, 프로필렌글리콜과 물의 혼합액으로 추출한 추출물이다. 또한 상기한 자연산 상황버섯 추출물은 피부화장료의 성분으로서 적용가능하며 이때 함유량은 0.05-30%인 것이 바람직하다.The present invention is to provide a skin cosmetic excellent in safety and excellent in the prevention and improvement effect of various skin damage by taking advantage of the above-mentioned problems of the prior art. Natural situation mushroom extract of the present invention for achieving the above object has a situation mushroom (P. pini and P. linteus) that has been grown in the natural state of the North Korean peninsula, hot water extract, 20-30% ethanol extract, 1 It is an extract extracted with a mixture of, 3-butylene glycol, propylene glycol and water. In addition, the above-mentioned natural situation mushroom extract is applicable as a component of the skin cosmetic, and the content is preferably 0.05-30%.

본 발명에서는 인공재배한 상황버섯이 아니고 북한지방의 자연상태서 성장한 자연산 상황버섯으로부터 열수추출한 추출물과, 에탄올 추출물, 부틸렌글리콜 추출물, 프로필렌글리콜과 이들의 혼합용매로 추출한 추출물에서 화장품에 이용할 수 있는 효과가 입증되고 화장품에 함유시 우수한 항산화효과, 미백효과 등이 발견되어 본 발명을 하게 된 것이다. 우선, 상황버섯 추출물의 피부화장료에 대한 이용가능성을 확인하기 위하여 미백효과, 항산화효과 등을 조사했다. 시료로 사용하기위해 자연산 상황버섯을 실온에서 중량의 변화가 없을 때까지 건조시킨 후,일반적인 기계적으로 분쇄하여 분말로 하였다. 이 분말 20g을 30% 프로필렌글리콜수용액 80g을 가하여 60℃에서 48시간 물중탕하여 추출한 후 0.45㎛ 여과지로 여과한 액과 상황바삿 20g을 화장품용으로 사용하는 30% 에탄올 수용액 80g에 실온에서 10일간 방치한 후 0.45㎛(㎛)로 제균여과한 것, 그리고 상황버섯 20g을 증류수 80g에 첨가하고 80℃에서 48시간 물중탕하여 처리한 후 400메쉬크기의 쇠그물망으로 버섯과 여과액을 분리하고 계속해서 O.45㎛ 제균여과지로 여과한것을 시료로 사용하였다. 자외선흡수스펙트럼 분석결과 상황버섯 추출물들은 강한 자외선흡수능력을 갖고 있었다. 또한, 피부화장료로 응용하기 위한 효과 테스트로 리놀산 자동산화를 이용한 항산화효과 테스트에서 상황버섯 에탄올 추출물은 항산화효과가 대조군인 인공재배한 상황버섯추출물(45%)과 비타민 이(42.3%)에 비해 우수한 항산화효과(58%)를 가진 것으로 나타났다.In the present invention, the effect that can be used in cosmetics from the extract of hot water extracted from natural mushrooms grown in the natural state of North Korea, ethanol extract, butylene glycol extract, propylene glycol and a mixed solvent thereof, not artificially grown situation mushrooms It has been proved and excellent antioxidant effect, whitening effect, etc. when contained in cosmetics is to make the present invention. First, the whitening effect and antioxidant effect were investigated to confirm the availability of the skin extracts of situational mushroom extracts. For use as a sample, natural mushrooms were dried at room temperature until there was no change in weight, and then pulverized into a general mechanical powder. 20 g of this powder was added with 80 g of 30% aqueous propylene glycol solution and extracted by boiling in water at 60 ° C for 48 hours. The solution was filtered with a 0.45 µm filter paper and 80 g of 30% ethanol aqueous solution using 20 g of ethanol for cosmetics. After sterile filtration to 0.45㎛ (㎛), 20g of the situation mushroom was added to 80g of distilled water and treated with water bath at 80 ℃ for 48 hours, and then the mushroom and filtrate were separated with a mesh mesh of 400 mesh size and then continued. The sample was filtered through a 45.5-micron sterile filter. As a result of UV absorption spectrum analysis, the situation mushroom extracts had strong UV absorption ability. In addition, in the antioxidant test using linoleic acid as an effect test for application to skin cosmetics, ethanol extract of the situation mushroom was superior to the artificial mushroom cultivated mushroom extract (45%) and vitamin I (42.3%), which were antioxidant controls. It was found to have antioxidant effect (58%).

티로시나제 활성 저해능, 미생물(균주명:Streptomyces bikiniensis)의 멜라닌 생성 저해능, 비16-에프1 멜라노마 세포의 멜라닌 생성 저해능 등의 실험으로 상황버섯 추출물에 대한 미백효과를 조사하였다. 그 결과 티로시나제 활성을 50% 저해하는 농도(IC50)는 0.3%이하였고,0,005%의 농도에서 미생물의 멜라닌 생성 영역을 25밀리미터 저해하였다. 또한 B16 -F1 멜라노마 세포의 멜라닌 생성을 저해하는 최소 농도는 0.05%로 나타나 피부외용제 특히 화장료로서 우수한 효과를 나타냈다. 본 발명의 자연산 상황버섯추출물을 함유한 피부화장료로서의 적용예는 현재 어느곳에도 없다. 이 자연산 상황버섯추출물의 피부화장료에의 배합량은 한정된 것은 아니지만 0.05중량%내지 30중량%가 적당한 것으로 나타났으며 너무 적은 함량에서는 효과를 기대하기 어렵고, 너무 많은 함량에서는 그 이상배합해도 효과가 크게 증가하지 않았다. 또한 본 발명의 추출물은 물에 쉽게 혼화되고 유지와 유기성 용매에는 혼화가 어렵지만 현탁상태에서도 피부화장료에 배합은 가능하고 효과에 대하여 어뗘한 영향도 주지 않는다. 본 발명의 피부외용제는 피부화장료로서 이용되는 것이 적당하며 일반 피부화장료에 배합되는 보통의 성분 예를들면 유분, 물, 계면활성제, 보습제, 저급알콜, 증점제, 킬레이트제, 색소, 방부제, 향료 등을 필요한 만큼 적용배합하는 것이 가능하다. 역시 본 발명에서 피부화장료는 피부에 사용하는 것을 널리 알리고 예를들면, 화장수, 유액, 크림,팩 등의 화장료와 특히 미백화장료에 응용이 대표적이다.The effect of inhibiting tyrosinase activity, inhibiting melanogenesis of microorganisms (Streptomyces bikiniensis), and inhibiting melanogenesis of non-16-F1 melanoma cells were investigated. As a result, the concentration (IC 50 ) that inhibits tyrosinase activity by 50% was less than 0.3%, and the melanin-producing region of the microorganism was inhibited by 25 mm at the concentration of 0,005%. In addition, the minimum concentration of inhibiting melanin production of B16-F1 melanoma cells was 0.05%, showing an excellent effect as a skin external preparation, especially a cosmetic. There is currently no example of application as a skin cosmetic containing the natural situation mushroom extract of the present invention. Although the amount of this natural mushroom extract to skin cosmetics is not limited, it is found that 0.05 to 30% by weight is appropriate, and the effect is not expected at too small content, and the effect is greatly increased even if it is mixed at too much content. Did not do it. In addition, the extract of the present invention is easily mixed with water and difficult to blend with fats and oils and organic solvents, but can be blended into the skin cosmetic even in a suspended state and does not have any influence on the effect. Skin external preparation of the present invention is suitable to be used as a skin cosmetic, and common ingredients such as oil, water, surfactants, moisturizers, lower alcohols, thickeners, chelating agents, pigments, preservatives, fragrances, etc. It is possible to combine them as necessary. Also in the present invention, the skin cosmetics are widely known for use on the skin, for example, the application of cosmetics such as lotion, latex, creams, packs and especially whitening cosmetics.

이하, 실시예에 의해 본 발명을 보다 상세히 설명한다.Hereinafter, the present invention will be described in more detail with reference to Examples.

이들 실시예는 본 발명의 예시적인 기재일 뿐이며 본 발명의 범위가 이들 실시예에 국한되는 것은 아니다.These examples are merely illustrative of the present invention and the scope of the present invention is not limited to these examples.

실시예1Example 1

상황버섯은 한반도의 북한 지방에서 자생한 낙엽진흙버섯(P. pini)과 목질진흙버섯(P.linteus)을 수입사용하였으며 시료로 사용하기위해 상황버섯을 실온에서 중량의 변화가 없을 때까지 건조시킨 후, 일반적인 기계적인 방법으로 분쇄하여 위두가지 버섯을 동량씩 혼합 사용하였다. 용매추출하기위해서 용매를 여러 가지로 선택하였는데, 증류수, 에탄올, 1,3-부틸렌글리콜, 프로필렌글리콜과 이들의 조합에 의한 것이다. 물에 의한 추출은 물에 20%되게 상황버섯분말을 넣고 60℃에서 48시간 물중탕한 후 400메쉬되는 쇠그물망으로 여과하여 버섯과 여과된 액을 분리하였다. 계속해서 분리된 여과액을 0.45㎛ 여과지로 제균여과하여 시료로 사용하였다, 에탄올에 의한 추출은 물에 대한 에탄을 30%용액에 상황버섯 20%되게 15일간 실온에 방치 한 후 400메쉬되는 그물망으로 여과하여 버섯과 여과액을 분리하고 이 분리된 여과액을 0.45㎛ 여과지로 여과하여 시료로 하고 다음의 실시예와 같은 효능효과시험을 하였다. 1,3-부틸렌, 프로필렌글리콜과 물의 혼합물에 의한 추출에서는 1,3-부틸렌, 프로필렌글리콜과 물의 비율을 8대2와 4대6의중량비로 혼합하고 이 혼합액에 20중량%되게 상황버섯을 첨가하고 60℃에서 48시간 가열처리한 후 400메쉬되는 쇠그물망으로 여과하여 버섯과 여과액을 분리한다음 여과액을 0.45㎛ 여과지로 여과하여 시료로 하였다.Situation mushrooms were imported P. pini and P. linteus native to North Korea on the Korean Peninsula. Situation mushrooms were dried at room temperature until there was no change in weight for sample use. After that, it was pulverized by a general mechanical method, and the two kinds of mushrooms were used in equal amounts. A variety of solvents were selected for solvent extraction, based on distilled water, ethanol, 1,3-butylene glycol, propylene glycol and combinations thereof. Extraction by water was added to the situation mushroom powder in water to 20% in a water bath at 60 ℃ 48 hours and then filtered through a mesh mesh of 400 mesh to isolate the mushroom and the filtered liquid. Subsequently, the separated filtrate was sterilized by 0.45 μm filter paper and used as a sample. The extraction by ethanol was carried out by leaving the ethane for water in a 30% solution for 15 days at room temperature with 20% of situation mushrooms, followed by a mesh of 400 mesh. The filtrate was separated from the mushroom and the filtrate. The separated filtrate was filtered with 0.45 μm filter paper to prepare a sample, and the same effect test as in the following example was performed. In the extraction by the mixture of 1,3-butylene, propylene glycol and water, the ratio of 1,3-butylene, propylene glycol and water is mixed in a weight ratio of 8 to 2 and 4 to 6, and 20% by weight of the situation mushroom The mixture was heated at 60 ° C. for 48 hours, filtered through a mesh mesh of 400 mesh, mushrooms and filtrates were separated, and the filtrate was filtered with 0.45 μm filter paper to prepare a sample.

실시예2Example 2

본 실시예는 자동산화가 잘 되는 물질인 리놀산을 이용하여 이 자동산화를 억제하는 정도를 상호비교한 것이다.8㎖ 캡튜브에 10mM 리놀산용액 3㎖와 4㎎/㎖로 조정한 샘플 0.075㎖를 첨가한 후 4℃에 방치한다. 상기의 혼합액 0.1㎖를 24시간 후에 75% 에탄올수용액 4.7㎖가 든 시험관에 첨가하여 교반한 후, 30% 티오시안산암모니움용액 0.1㎖와 염화제1철 시약 0.l㎖를 첨가하고 3분간 방치한후 발색시켜 500nm에서 흡수파장을 측정하여 물질의 항산화효과 시료를 넣지않은 대조군과 비교하여 항산화효과를 %로 측정했다. 리놀산 자동산화를 이용한 항산화효과 시험에서 실시예1에 의해 추출한 자연산 상황버섯 추출물은 항산화효과가 대조군에 비해 뛰어났으며 비타민 E(42.3%)에 비해 우수한 항산화효과(58%)를 가진 것으로 나타났다(표 1).In this example, the degree of inhibition of this automatic oxidation was compared using linoleic acid, which is a well-oxidized material. In an 8 ml captube, 3 ml of 10 mM linoleic acid solution and 0.075 ml of sample adjusted to 4 mg / ml were prepared. After addition, it is left at 4 ° C. 0.1 ml of the mixed solution was added to a test tube containing 4.7 ml of 75% ethanol aqueous solution after 24 hours, followed by stirring. Then, 0.1 ml of 30% ammonium thiocyanate solution and 0.1 ml of ferrous chloride reagent were added thereto for 3 minutes. After leaving, color was absorbed and the absorption wavelength was measured at 500 nm. The antioxidant effect of the substance was measured in% compared to the control group without the sample. In the antioxidant effect test using linoleic acid automatic oxidation, the natural situation mushroom extract extracted in Example 1 was superior to the control group and showed superior antioxidant effect (58%) compared to vitamin E (42.3%). One).

표1Table 1

실시예3Example 3

본 실험은 실시예1에 의해 추출한 상황버섯 추출물의 미백효과를 알아보기위해 티로시나제라는 효소가 기능억제되는 정도를 보고 미백효과를 판단한 것이다. 티로시나제는 생체내에서 티로신이라는 물질이 산화되는 과정을 촉진하여 멜라닌이 생성되게 도와주는 효소이다. 본 실시예에서는 이 효소(시그마(미국)), 의 기능을 억제하여 티로신이 산화되어 멜라닌이라는 흑색의 고분자를 형성하는 것을 억제하는 정도를 보는 방법(Pomerantz S. H., The tyrosine hydroxylase activity of mammalian tyrosinase, J. Biochem.,24,16l-l68(1966)) 을 응용해 미백효과로 판정하였다. 상백피 추출물 (원액, 10, 1, 0.1, 0.01의 증류수에 대한 부피/부피 %),자연산, 인공산 상황버섯 추출물 (원액, 0.5, 0.05, 0.005, 0,0005의 증류수에 대한 부피/부피 %),고직산, 알부틴, 하이드로퀴논, 피-메톡시페놀(1, 0.1, 0.01, 0.001,0.0001의 증류수에 대한 부피/부피 %)을 시료로 사용하여 티로시나제 활성 억제효과를 조사하였다.This experiment was to determine the whitening effect by looking at the degree of functional inhibition of the enzyme tyrosinase to determine the whitening effect of the situation mushroom extract extracted in Example 1. Tyrosinase is an enzyme that helps melanin production by promoting the oxidation of a substance called tyrosine in vivo. In this embodiment, the method of inhibiting the function of this enzyme (Sigma (USA)), to inhibit the tyrosine oxidized to form a black polymer called melanin (Pomerantz SH, The tyrosine hydroxylase activity of mammalian tyrosinase, J) Biochem., 24, 16l-l68 (1966)) was applied as a whitening effect. Morus extract (volume / volume% for distilled water of stock solution, 10, 1, 0.1, 0.01), natural acid, artificial acid situation mushroom extract (volume / volume for distilled water of stock solution, 0.5, 0.05, 0.005, 00005) Inhibition of tyrosinase activity was investigated by using a sample of Gojic Acid, Arbutin, Hydroquinone, and P-methoxyphenol (volume / volume% of distilled water of 1, 0.1, 0.01, 0.001, 0.0001) as a sample.

티로시나제에 대한 저해활성은 시료 15㎕를 마이크로플레이트(96웰)에 넣고, 0.1몰 인산완충액(pH6.86) 150㎕ 1.5mM 엘-티로신용액 25㎕를 넣은 후,2,380유니트/㎖ 머쉬룸 티로시나제(0.05mol 인산완충액, pH6.86) 7㎕를 첨가하여 30℃에서 10분간 반응시킨 후 마이크로플레이트를 사용하여 490nm에서 측정하였다. 티로시나제에 대한 저해율(%)은 다음식에 의하여 계산하였으며,IC50값은 효소활성 저해율 50%에 달하는 저해물질의 농도로 결정하였다(표2).For inhibitory activity against tyrosinase, 15 μl of the sample was placed in a microplate (96 wells), 150 μl of 0.1 mol phosphate buffer (pH6.86) and 25 μl of 1.5 mM L-tyrosine solution were added, followed by 2,380 unit / ml mushroom tyrosinase (0.05). mol phosphate buffer, pH6.86) 7 μl was added and reacted at 30 ° C. for 10 minutes, and measured at 490 nm using a microplate. Inhibition rate (%) for tyrosinase was calculated by the following equation, IC 50 value was determined by the concentration of inhibitors reaching 50% enzyme inhibition rate (Table 2).

저해율(%) = {[(D-C)-(B-A)] / (D-C)} × 100% Inhibition = {[(D-C)-(B-A)] / (D-C)} × 100

A : 저해제를 넣은 것의 반응 전 흡광도A: absorbance before reaction of the inhibitor

B : 저해제를 넣은 것의 반응 후 흡광도B: absorbance after the reaction of the inhibitor

C : 저해제를 넣지 않은 것의 반응 전 흡광도C: Absorbance before reaction of without inhibitor

D : 저해제를 넣지 않은 것의 반응 후 흡광도D: absorbance after the reaction without the inhibitor

티로시나제 활성 억제작용시험결과 자연산 상황버섯 각 추출물의 IC50값은 0.2%이하로 나타났다. 이는 인공재배한 상황버섯보다 효과가 높았으며 기존에 미백효과가 우수한 것으로 알려지고 있는 알부틴보다도 우수한 효과를 가지는 것으로 나타났다.As a result of tyrosinase activity inhibition test, the IC 50 value of each extract of wild situation mushroom was less than 0.2%. This effect was higher than artificial cultivated situation mushrooms and had better effects than arbutin, which is known to have excellent whitening effect.

표2Table 2

실시예4Example 4

본 실험은 방선균에 대한 멜라닌 생성 억제작용을 측정한 것으로 사용된 균주(Streptomyces bikiniensis NRRL B-1049)는 KCTC(Korean Collection for Type Culture, 기탁번호 9172)로부터 분양받아 사용하였다.This experiment was used to measure the melanin inhibitory activity against actinomycetes (Streptomyces bikiniensis NRRL B-1049) was used from KCTC (Korean Collection for Type Culture, Accession No. 9172).

Streptomyces bikiniensis NRRL B-1049를 papavizas′ VDYA agar slant 배지(V-8 juice 2O0㎖, glucose 2g, yeast extract 2g, CaCO31g, agar 20g, 증류수 800㎖, pH7.2)에서 2주간 28℃로 배양시켜 포자를 생성시킨 후 멸균수로 포자 현탁액을 만들었다. 0.2%의 yeast extract를 첨가한 ISP No.7 평판배지에 포자현탁액 0.2㎖씩을 도포한 후 배지표면에 시료를 적신 페이퍼 디스크에 시료양을 30㎍/paper disc로 도포하고 28℃에서 배양하였다. 48시간 배양 후 생성된 멜라닌 생성 저해환의 크기를 멜라닌생성 저해물질로 알려진 4-하이드록시아니솔을 대조구로 하여 멜라닌생성 저해여부를 관찰하였다(표3).Incubate Streptomyces bikiniensis NRRL B-1049 at 28 ° C for 2 weeks in papavizas ′ VDYA agar slant medium (V-8 juice 2O0ml, glucose 2g, yeast extract 2g, CaCO 3 1g, agar 20g, distilled water 800ml, pH7.2) Spores were produced to produce spores with sterile water. 0.2 ml each of the spore suspension was applied to ISP No. 7 plate medium to which 0.2% yeast extract was added, and then the amount of the sample was applied to the paper disc soaked on the surface of the medium with 30 ㎍ / paper disc and incubated at 28 ° C. The inhibition of melanogenesis was observed using the 4-hydroxyanisole known as the melanogenesis inhibitor as a control for the size of the melanogenesis inhibitors produced after 48 hours of incubation (Table 3).

자연산 상황버섯 각 추출물의 Streptomyces bikiniensis NRRL B-1049에 대한 melanin 생성저해활성은 자연산 에탄올 추출물의 경우 30mm의 멜라닌 생성저해환을 나타내었다· 이러한 저해활성은 강력한 멜라닌 생성저해제인 4-히드록시아니솔보다 우수한 활성을 나타내었다.The inhibitory activity of melanin-producing activity against Streptomyces bikiniensis NRRL B-1049 of wild-state extracts showed 30 mm melanin-inhibiting activity in natural ethanol extracts. This inhibitory activity was higher than that of 4-hydroxyanisole, a potent melanin-inhibiting agent Excellent activity was shown.

표3Table 3

실시예4Example 4

본 실험은 가연산 상황버섯의 열수추출물, 에탄올 추출물,1,3-부틸렌글리콜, 프로필렌글리콜과 물의 혼합액으로 추출한 추출물의 피부미백효과를 알아보기위해 세포수준에서 관찰한 것이다.This experiment was observed at the cellular level to investigate the skin whitening effect of the extract extracted with hot water extract, ethanol extract, 1,3-butylene glycol, propylene glycol and water mixture of citric acid situation mushroom.

본 실험에 사용된 B16-Fl 멜라노마세포는 마우스에서 유래한 세포균주이며 멜라닌이라는 혹색색소를 분배하는 세포이다. 이 세포를 인공배양중에 물질을 처리하여 멜라닌흑색색소가 감소하는정도를 비교평가하였다.The B16-Fl melanoma cells used in this experiment are cell strains derived from mice and are cells that distribute a melanin called melanin. The cells were treated with material during artificial culture and the degree of melanin black pigment reduction was compared.

본 실험에 사용된 B16-F1 멜라노마세포는 ATCC(American Type Culture Collection, 기탁번호:6323)로부터 분양받아 사용하였다.B16-F1 melanoma cells used in this experiment were used from the American Type Culture Collection, Accession No .: 6323 (ATCC).

B16 멜라노마 세포의 멜라닌 생합성 저해도 판별은 다음과 같이 행하였다. B16멜라노마 세포를 5×103cel1s/㎖ 농도로 10% 소태아혈청을 포함하는 Dulbecco's Modified Eagle's Medium(Gibco, USA)에 현탁시켰다. 현탁세포(5㎖)를 조직배양 플라스크에 넣은 후 검정 시료를 0.134% 첨가한 후, 5% 이산화탄소-95% 공기조건으로 37℃에서 배양하였다. 4일간 배양후 배양액의 색으로 멜라닌생성 정도를 l차 판별하고, 세포를 인산완충액으로 씻은 후 트립신처리하여 세포를 플라스크로부터 분리하였다. 세포를 튜브에 모은 후 인산완충액으로 씻고, 원심분리하여 튜브 아래쪽에 침전된 멜라닌 생성양을 기존의 미백제로 알려지고 있는 몇개의 시료와 상황버섯추출물과의 멜라닌색소분비억제정도를 비교한 것이다. 평가방법은 세포배양중에 아무것도 처리하지 않은 것과 각 시료를 처리했을 때의 검은 멜라닌색소 분비정도에 따라 색상이 변화되는 정도를 육안관찰하여 상호비교 평가하였다. 결과적으로 자연산 상황버섯추출물이 멜라닌색소분비를 효과있게 억제한다는 결과를 얻었다(표4).The inhibition of melanin biosynthesis of B16 melanoma cells was determined as follows. B16 melanoma cells were suspended in Dulbecco's Modified Eagle's Medium (Gibco, USA) containing 10% fetal bovine serum at a concentration of 5 × 10 3 cels / ml. Suspension cells (5 ml) were placed in a tissue culture flask, and 0.134% of the assay sample was added, followed by incubation at 37 ° C under 5% carbon dioxide-95% air. After incubation for 4 days, the melanogenesis level was first determined by the color of the culture medium, and the cells were washed with phosphate buffer and trypsinized to separate the cells from the flask. The cells were collected in a tube, washed with phosphate buffer, and centrifuged to compare melanin secretion levels between several samples known as whitening agents and situation mushroom extracts. The evaluation method was evaluated by visual observation of the degree of color change according to the treatment of nothing in the cell culture and the degree of black melanin pigment secretion when each sample was treated. As a result, natural situation mushroom extract effectively inhibited melanin pigment secretion (Table 4).

표4.Table 4.

실시예 5,6,7Examples 5,6,7

본 실시예에서는 실시예1에 의해 추출한 각각의 자연산 상황버섯 추출물을 함유한 화장료의 비교실험을 행하였다. 비교실험에 사용된 크림의 조성은 표5에 나타내었다. 우선 표5에 기륵되어 있는 나)상을 가열하여 70℃에 보존한다. 이것에 가)상을 가하여 예비유화 후 호모믹서로 균일하게 유화하고 다음에 서서히 냉각하여 크림(실시예 5-7, 비교예)을 조제한다. 표5의 각 크림을 실험자(20세 -35세의 여성)5명의 얼굴오른쪽에 1일 2회 연속 1개월간 도포하였다. 실험완료 후 얼굴 좌우 양편의 도포부위 피부를 영상분석기로 얼굴색을 비교하여 가장 어두운색을 5 중간색을 3, 가장 환한 색을 1로 정하고 그 중간정도를 어림잡아 평가하었다. 표6은 제5실시예로 만든 크림을 사용한 실험자의 안면피부색을 비교예로 만든 크림을 사용한 실험자의 안면사용부위와 비교한 것이다. 표6에서와 같이 상황추출물을 함유한 크림을 도포한 실험자의 안면에서 미백효과가 나타난 것을 알수 있다.In this example, a comparative experiment of the cosmetics containing the natural situation mushroom extract extracted in Example 1 was conducted. The composition of the cream used in the comparative experiment is shown in Table 5. First, the phase b) described in Table 5 is heated and stored at 70 ° C. To this, a) phase is added, and after pre-emulsification, it is emulsified uniformly with a homomixer, and it cools gradually, and prepares a cream (Example 5-7, a comparative example). Each cream of Table 5 was applied to the right side of five subjects (20-35 year old female) twice a day for 1 consecutive month. After completion of the experiment, the skins of the applied areas on the left and right sides of the face were compared by using an image analyzer, and the darkest color was set as 5 medium color and 3 as the brightest color. Table 6 compares the facial skin color of the experimenter using the cream made in Example 5 with the facial use area of the experimenter using the cream made of the comparative example. As shown in Table 6, it can be seen that the whitening effect appeared on the face of the experimenter who applied the cream containing the situation extract.

표5Table 5

i i

표6Table 6

이하에 그 외의 실시예를 나타내었다. 즉, 앞에서와 같이 피부개선에 우수한 효과를 나타내었으며 피부미용효과를 나타내었다.Other examples are shown below. That is, as shown above, it showed an excellent effect on skin improvement and a skin beauty effect.

실시예8Example 8

95% 에탄올 8g에 폴리피로리돈 0.05g, 올레일알콜 0.1g, 폴리옥시에틸렌모노올레이트 0.2g, 향료 0.2g, 파라옥시안식향산메릴에스테르 0.1g, 소량의 산화방지제, 소량의 색소를 혼합용해한다. 이것을 자연산 상황버섯 추출물0.05g, 글리세린 5g을 정제수 85.33g에 용해한 것 중에 교반첨가하여 피부개선효과가 있는 화장수를 얻는다.To 8 g of 95% ethanol, 0.05 g of polypyrrolidone, 0.1 g of oleyl alcohol, 0.2 g of polyoxyethylene monooleate, 0.2 g of fragrance, 0.1 g of paraoxybenzoic acid meryl ester, a small amount of antioxidant, and a small amount of pigment are dissolved. . This was added with stirring in a solution of 0.05 g of natural situation mushroom extract and 5 g of glycerin in 85.33 g of purified water to obtain a lotion having a skin-improving effect.

실시예9Example 9

세틸알콜 1.2g, 스쿠알란 10g, 바세린 2g, 파라옥시안식향산에틸에스테르 0.2g, 글리세린모노에스테아레이드 1g, 폴리옥시에틸렌(20mol부가)모노올레이트 1g 및 향료 0.1g을 70℃에서 가열혼합용해하여 자연산 상황버섯추출물 0.5g, 디프로필렌글리콜 5g, 폴리에틸렌글리콜1500 2g, 트리에탄올아민 0.2g, 정제수 76.2g을 75℃로 가열해서 가열 용해시킨다. 양자를 혼합하여 유화시킨 후 냉각하여 수/유중계형의 피부개선효과가 있는 유액을 얻는다.1.2 g of cetyl alcohol, 10 g of squalane, 2 g of vaseline, 0.2 g of paraoxybenzoic acid ethyl ester, 1 g of glycerin monoesteralide, 1 g of polyoxyethylene (20 mol added) monooleate and 0.1 g of fragrance at 70 ° C. 0.5 g of a situation mushroom extract, 5 g of dipropylene glycol, 2 g of polyethylene glycol 1500, 0.2 g of triethanolamine, and 76.2 g of purified water are heated to 75 ° C. to dissolve it by heating. Both are emulsified by mixing, followed by cooling to obtain an emulsion having a water / oil-based skin improvement effect.

실시예10Example 10

95% 에틸알콜 5g에 폴리옥시에틸렌 솔비탄모노올레이트 1.2g, 해조추출물 0.3g, 히야론산나트륨 0.2g, 비타민이 아세테이트 0.2g, 감초산나트륨 0.2g, 파라옥시안식향산에틸에스테르 0.1g, 자연산 상황버섯추출물 1g 및 적량의 색소를 혼합하여 피부개선의 효과가 있는 미용액을 얻는다.5 g of 95% ethyl alcohol, 1.2 g of polyoxyethylene sorbitan monooleate, 0.3 g of seaweed extract, 0.2 g of sodium hyaluronate, 0.2 g of vitamin acetate, 0.2 g of sodium licorice, 0.1 g of paraoxybenzoate, 0.1 g of natural products 1g of mushroom extract and a suitable amount of pigment are mixed to obtain a cosmetic liquid having an effect of skin improvement.

본 발명에 의한 자연산 상황버섯추출물은 각종 피부개선효과가 탁월하며 안전성면에서도 매우 우수하다. 또한, 상술한 자연산 상황버섯 추출물은 피부화장료에 적용이 가능하며 화장료에 함유시 미백효과와 항산화효과가 우수하다.Natural situation mushroom extract according to the present invention is excellent in various skin improvement effect and very excellent in terms of safety. In addition, the above-mentioned natural situation mushroom extract can be applied to skin cosmetics and has excellent whitening and antioxidant effects when contained in cosmetics.

Claims (6)

자연산 상황버섯(P. pini와 P. linteus))추출물을 함유한 피부화장료를 특징으로 하는 것Characterized by skin cosmetics containing natural P. pini and P. linteus extracts 건조, 분쇄한 자연산 상황버섯을 1,3-부틸렌글리콜, 프로필렌글리콜과 증류수의 혼합액으로 추출한 것을 특징으로 하는 것.The dried and pulverized natural situation mushroom is extracted with a mixed solution of 1,3-butylene glycol, propylene glycol and distilled water. 건조한 상황버섯을 30% 내지 95% 에탄올수용액으로 추출 한 것을 특징으로 하는 것.The dried situation mushroom, characterized in that extracted with 30% to 95% ethanol aqueous solution. 건조한 자연산 상황버섯을 증류수로 열수추출한것을 특징으로 하는 것.It is characterized by the extraction of hot natural dry mushrooms with distilled water. 제2항의 1,3-부틸렌글리콜, 프로필렌글리콜과 증류수의 혼합액중 1,3-부틸렌글리콜, 프로필렌글리콜과 증류수의 혼합비율이 8대2내지 4 대6인 것을 특징으로 하며 추출시 온도를 실온 내지 60℃로 가열추출한 후 제균여과한 것을 특징으로 하는 것.The mixing ratio of 1,3-butylene glycol, propylene glycol and distilled water in the mixed solution of 1,3-butylene glycol, propylene glycol and distilled water of claim 2 is characterized in that the temperature at the time of extraction After sterilizing filtration after heating to room temperature to 60 ℃. 제1항의 자연산 상황버섯 추출물을 0.05내지 30%를 함유하는 것을 특징으로 하는 피부화장료Skin cosmetics, characterized in that it contains 0.05 to 30% of the natural situation mushroom extract of claim 1
KR1019980015036A 1998-04-28 1998-04-28 Skin cosmetic using natural mushrooms KR19990081226A (en)

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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR19990046601A (en) * 1999-04-02 1999-07-05 김준, 김미란 Soap including Phellinus Linteus as the active component
KR100365242B1 (en) * 2000-10-17 2002-12-18 최찬기 composite of cosmetics contained chondroitin and pellinus linteus extract
KR20040015469A (en) * 2002-08-13 2004-02-19 주식회사 래디안 Cosmetic composition containing extract from phellinus linteus cultivated in rice
KR20040043971A (en) * 2002-11-20 2004-05-27 동방에프티엘 주식회사 Extract compositions comprising a natural phellinus linteus, green tea, caryophilli flos and rosae fructus and cosmetic products comprising their extracts
KR100538536B1 (en) * 2002-11-22 2005-12-22 주식회사 다사랑 The manufacturing method of the beauty pack including salicornia herbaceea.
KR100695996B1 (en) * 2001-02-20 2007-03-15 나드리화장품주식회사 Niosome containing natural phellinus linteus extract and cosmetic composition containing it
KR101662272B1 (en) * 2015-03-24 2016-10-05 상황미인(주) Natural cosmetic composition for skin moisturizing comprising Mixed extract of barley and germ and rice bran of black rice and preparation method thereof

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR19990046601A (en) * 1999-04-02 1999-07-05 김준, 김미란 Soap including Phellinus Linteus as the active component
KR100365242B1 (en) * 2000-10-17 2002-12-18 최찬기 composite of cosmetics contained chondroitin and pellinus linteus extract
KR100695996B1 (en) * 2001-02-20 2007-03-15 나드리화장품주식회사 Niosome containing natural phellinus linteus extract and cosmetic composition containing it
KR20040015469A (en) * 2002-08-13 2004-02-19 주식회사 래디안 Cosmetic composition containing extract from phellinus linteus cultivated in rice
KR20040043971A (en) * 2002-11-20 2004-05-27 동방에프티엘 주식회사 Extract compositions comprising a natural phellinus linteus, green tea, caryophilli flos and rosae fructus and cosmetic products comprising their extracts
KR100538536B1 (en) * 2002-11-22 2005-12-22 주식회사 다사랑 The manufacturing method of the beauty pack including salicornia herbaceea.
KR101662272B1 (en) * 2015-03-24 2016-10-05 상황미인(주) Natural cosmetic composition for skin moisturizing comprising Mixed extract of barley and germ and rice bran of black rice and preparation method thereof

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