KR102415369B1 - Functional bath cleanser composition comprising plant powder - Google Patents
Functional bath cleanser composition comprising plant powder Download PDFInfo
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- KR102415369B1 KR102415369B1 KR1020200155527A KR20200155527A KR102415369B1 KR 102415369 B1 KR102415369 B1 KR 102415369B1 KR 1020200155527 A KR1020200155527 A KR 1020200155527A KR 20200155527 A KR20200155527 A KR 20200155527A KR 102415369 B1 KR102415369 B1 KR 102415369B1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- 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
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- A—HUMAN NECESSITIES
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- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- 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
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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- A—HUMAN NECESSITIES
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Abstract
본 발명은 녹차 1~40 중량%, 병풀 1~20 중량%, 쑥 1~30 중량%, 생강 1~10 중량% 등의 식물 성분을 포함함으로써 항산화, 미백, 주름 개선, 항염증, 상처치료 및 보습 등의 기능성을 가지는 식품 조성물 및 화장료 조성물, 특히 입욕제 조성물을 개시한다.The present invention provides antioxidant, whitening, wrinkle improvement, anti-inflammatory, wound healing and Disclosed are a food composition and a cosmetic composition having a function such as moisturizing, particularly a bath composition.
Description
본 발명은 식물 분말 또는 식물 추출물을 포함하는 조성물에 관한 것이다. 구체적으로는 식물 분말 또는 식물 추출물을 주성분으로 포함하는 화장료 조성물, 식품 조성물에 관한 것으로, 특히 기능성 입욕제 조성물에 관한 것이다.The present invention relates to a composition comprising a plant powder or plant extract. Specifically, it relates to a cosmetic composition or a food composition comprising a plant powder or a plant extract as a main component, and more particularly, to a functional bath composition.
현대인들은 환경오염 물질의 노출, 스트레스, 색조화장에 따른 잦은 세안, 연령 증가에 따른 자연적인 피부 노화 등의 여러 가지 원인으로 인하여 피부 건조, 피부 각질 형성 등의 피부 손상에 쉽게 노출되고 있다. 따라서 피부의 보호 및 개선을 위한 노력이 요구되고 있다.Modern people are easily exposed to skin damage such as skin dryness and keratin formation due to various causes such as exposure to environmental pollutants, stress, frequent washing with color makeup, and natural skin aging due to increasing age. Therefore, efforts to protect and improve the skin are required.
다양한 스트레스로 세포내에서 염증이 유발하게 되면, 활성종의 하나인 NO (Nitric Oxide)가 발생하게 된다. 이는 피부 및 세포의 손상을 유발하고 노화를 유도하는 것으로 알려져 있다. 염증과 밀접한 관련이 있는 호중구는 피부에 염증이 발생되면 진피층으로 이동하여 엘라스타제를 분비하게 된다. 분비된 엘라스타제는 피부의 탄력에 관여하는 엘라스틴을 분해하여 주름을 유발시킨다. 또한 피부에서 염증이 발생하게 되면 염증매개 물질들에 의해 타이로시나아제(tyrosinase) 활성 증가를 통하여 과색소형성이 일어나게 된다. 피부 보습기능에 관여하는 히알루로니다아제(Hyaluronidase)는 히알루론산(hyaluronic acid)을 가수분해하는 효소로써 히알루로니다아제의 활성은 피부 건조의 원인이 될 수 있다.When inflammation is induced in cells by various stresses, NO (Nitric Oxide), one of the active species, is generated. It is known to cause damage to the skin and cells and induce aging. Neutrophils, which are closely related to inflammation, migrate to the dermal layer and secrete elastase when the skin becomes inflamed. The secreted elastase causes wrinkles by decomposing elastin, which is involved in the elasticity of the skin. In addition, when inflammation occurs in the skin, hyperpigmentation occurs through an increase in tyrosinase activity by inflammatory mediators. Hyaluronidase involved in the skin moisturizing function is an enzyme that hydrolyzes hyaluronic acid, and the activity of hyaluronidase may cause skin dryness.
입욕제는 유효물질을 적당한 농도로 배합해 욕탕에 넣은 뒤 입욕했을 때 단순히 피로 회복이나 피부 보호뿐만 아니라 질병치료 및 증상 완화의 효과까지 얻을 수 있는 약재를 말한다. 사람들은 고대부터 천연의 온천을 상처의 치료, 건강증진에 이용하였으며, 또한 각종 천연 식물을 활용하여 피부 건강의 목적으로 활용하여 왔다. 최근 들어 반신욕, 아로마 목욕 등의 목욕이 건강에 도움이 된다고 알려지면서 다양한 천연 식물성분을 지닌 입욕제를 이용한 목욕요법이 큰 관심을 끌고 있다. 그러나 화학성분이 주로 포함된 입욕제 제품은 다양하나, 식물성분으로 구성되어 제조되는 입욕제는 다양하지 않다. 따라서 식물성분으로 구성되어 부작용이 없으면서도 항산화, 항염증, 상처치료, 보습 등에 효과가 있는 입욕제 조성물의 개발이 필요하다.A bath agent refers to a drug that can not only recover from fatigue or protect the skin, but also treat diseases and relieve symptoms when taking a bath after mixing active substances in an appropriate concentration. People have used natural hot springs for wound healing and health promotion since ancient times, and various natural plants have been used for skin health purposes. Recently, as baths such as half-body baths and aroma baths are known to be beneficial to health, bath therapy using bath products containing various natural plant ingredients is attracting great attention. However, although there are various bath products containing mainly chemical components, bath products manufactured with plant components are not diverse. Therefore, there is a need to develop a bath composition that is effective in antioxidant, anti-inflammatory, wound healing, moisturizing, etc.
상기와 같은 문제점을 해결하기 위해 본 발명은 녹차, 병풀, 쑥, 생강 분말 성분의 배합비를 통하여 미백, 주름 개선 및 항염 효과를 제공하는 입욕제 조성물을 제공하는 것을 목적으로 한다.In order to solve the above problems, an object of the present invention is to provide a bath composition that provides whitening, anti-wrinkle and anti-inflammatory effects through a blending ratio of green tea, centella asiatica, mugwort, and ginger powder.
상기와 같은 목적을 달성하기 위해 본 발명은 녹차 1~40 중량%, 병풀 1~20 중량%, 쑥 1~30 중량%, 생강 1~10 중량%을 유효성분으로 함유하는 조성물을 제공한다.In order to achieve the above object, the present invention provides a composition containing 1 to 40% by weight of green tea, 1 to 20% by weight of Centella asiatica, 1 to 30% by weight of mugwort, and 1 to 10% by weight of ginger as active ingredients.
또한, 본 발명은 녹차, 병풀, 쑥, 생강 추출물을 유효성분으로 포함하는 주름개선 및 항염 효과를 위한 화장료 조성물 또는 식품 조성물을 제공한다.In addition, the present invention provides a cosmetic composition or food composition for wrinkle improvement and anti-inflammatory effect comprising green tea, centella asiatica, mugwort, and ginger extract as an active ingredient.
본 발명에 따른 녹차, 병풀, 쑥, 생강 분말을 유효성분으로 포함되어 있는 입욕제 조성물은 DPPH radical 소거활성 억제; 엘라스타아제(elastase)효소 활성 저해; 타이로시나아제 활성 저해; 세포 생존율 측정, nitric oxide (NO) 생성량 측정을 통하여 우수한 주름개선, 항염증 활성, 상처의 예방, 개선 또는 치료 효과, 보습 효과 등을 확인하였는바, 주름개선 및 피부 염증성 질환의 예방, 개선 또는 치료하는데 이용될 수 있다.The bath composition comprising green tea, centella asiatica, mugwort, and ginger powder according to the present invention as an active ingredient inhibits DPPH radical scavenging activity; inhibition of elastase enzyme activity; inhibition of tyrosinase activity; Excellent wrinkle improvement, anti-inflammatory activity, wound prevention, improvement or treatment effect, moisturizing effect, etc. were confirmed through cell viability measurement and nitric oxide (NO) production measurement. Prevention, improvement or treatment of wrinkles and skin inflammatory diseases can be used to
도 1은 본 발명에 따른 조성물의 DPPH 소거능 평가 결과를 나타낸 것이다.
도 2는 본 발명에 따른 조성물의 타이로시나제 활성저해 평가 결과를 나타낸 것이다.
도 3은 본 발명에 따른 조성물의 엘라스테이즈 활성 저해 평가 결과를 나타낸 것이다.
도 4는 본 발명에 따른 조성물의 염증 억제 평가 결과를 나타낸 것이다.
도 5는 본 발명에 따른 조성물의 상처의 예방 또는 개선 효과 평가 결과를 나타낸 것이다.
도 6는 본 발명에 따른 조성물의 보습 효과 결과를 나타낸 것이다.1 shows the evaluation results of DPPH scavenging ability of a composition according to the present invention.
2 shows the evaluation results of tyrosinase activity inhibition of the composition according to the present invention.
3 shows the evaluation results of the elastase activity inhibition of the composition according to the present invention.
4 shows the results of the evaluation of inhibition of inflammation of the composition according to the present invention.
5 shows the evaluation results of the prevention or improvement effect of the composition according to the present invention.
6 shows the moisturizing effect results of the composition according to the present invention.
이하, 본 발명을 하나의 구현예로서 더욱 상세하게 설명하면 다음과 같다.Hereinafter, the present invention will be described in more detail as one embodiment as follows.
본 발명은 녹차 분말, 병풀 분말, 쑥 분말, 생강 분말을 유효성분으로 함유하는 조성물을 제공한다.The present invention provides a composition comprising green tea powder, centella asiatica powder, mugwort powder, and ginger powder as active ingredients.
본 발명에 따른 녹차 분말, 병풀 분말, 쑥 분말, 생강 분말을 포함하는 조성물 또는 녹차 추출물, 병풀 추출물, 쑥 추출물, 생강 추출물을 포함하는 조성물은 입욕제가 피부에 대한 부적절한 반응과 및 피부 질환과 관련된 문제를 본질적으로 해결하거나 도움을 주는 화장료 조성물이나 식품 조성물로서 유용하다. According to the present invention, the composition comprising green tea powder, centella asiatica powder, mugwort powder, ginger powder, or the composition comprising green tea extract, centella asiatica extract, mugwort extract, and ginger extract, has problems related to the inappropriate reaction of the bathing agent to the skin and skin diseases. It is useful as a cosmetic composition or a food composition that essentially solves or helps.
본 발명의 바람직한 구현예에 따르면, 상기 조성물은 주름 개선 및 항염을 위한 유효성분으로 함유되는 입욕제 조성물로 사용될 수 있다According to a preferred embodiment of the present invention, the composition can be used as a bath composition contained as an active ingredient for wrinkle improvement and anti-inflammatory
본 발명의 바람직한 구현예에 따르면, 상기 화장료 조성물이나 식품 조성물은 통상적으로 화장료 조성물, 식품 조성물에 사용되는 첨가제나 부형제 또는 다른 효능을 가진 성분과 혼합된 형태로 이루어질 수 있다. According to a preferred embodiment of the present invention, the cosmetic composition or the food composition may be in a mixed form with additives or excipients or other ingredients that are typically used in cosmetic compositions and food compositions.
본 발명에 따른 화장료 조성물은 녹차(또는 녹차 추출물) 1~40 중량%, 병풀(또는 병풀 추출물) 1~20 중량%, 쑥(또는 쑥 추출물) 1~30 중량%, 생강(또는 생강 추출물) 1~10 중량%을, 바람직하게는 녹차 30~40 중량%, 병풀 20~30 중량%, 쑥 20~30 중량%, 생강 10~20 중량%로 함유할 수 있다.The cosmetic composition according to the present invention contains 1 to 40% by weight of green tea (or green tea extract), 1 to 20% by weight of Centella asiatica (or Centella asiatica extract), 1 to 30% by weight of mugwort (or mugwort extract), ginger (or ginger extract) 1 ~10% by weight, preferably 30-40% by weight of green tea, 20-30% by weight of Centella asiatica, 20-30% by weight of mugwort, 10-20% by weight of ginger.
본 발명의 일 실시예에서 상기 화장료 조성물은 녹차(또는 녹차 추출물) 40 중량%, 병풀(또는 병풀 추출물) 20 중량%, 쑥(또는 쑥 추출물) 30 중량%, 생강(또는 생강 추출물) 10 중량%로 함유할 수 있다. In an embodiment of the present invention, the cosmetic composition comprises 40% by weight of green tea (or green tea extract), 20% by weight of Centella asiatica (or Centella asiatica extract), 30% by weight of mugwort (or mugwort extract), 10% by weight of ginger (or ginger extract) may contain
본 발명에 따른 식품 조성물의 경우 녹차(또는 녹차 추출물) 1~40 중량%, 병풀(또는 병풀 추출물) 1~20 중량%, 쑥(또는 쑥 추출물) 1~30 중량%, 생강(또는 생강 추출물) 1~10 중량%을, 바람직하게는 녹차 30~40 중량%, 병풀 20~30 중량%, 쑥 20~30 중량%, 생강 10~20 중량%로 함유할 수 있다.In the case of the food composition according to the present invention, 1 to 40% by weight of green tea (or green tea extract), 1 to 20% by weight of Centella asiatica (or Centella asiatica extract), 1 to 30% by weight of mugwort (or mugwort extract), ginger (or ginger extract) 1 to 10% by weight, preferably 30 to 40% by weight of green tea, 20 to 30% by weight of Centella asiatica, 20 to 30% by weight of mugwort, and 10 to 20% by weight of ginger.
본 발명의 일 실시예에서 상기 식품 조성물은 녹차(또는 녹차 추출물) 40 중량%, 병풀(또는 병풀 추출물) 20 중량%, 쑥(또는 쑥 추출물) 30 중량%, 생강(또는 생강 추출물) 10 중량%로 함유할 수 있다.In an embodiment of the present invention, the food composition comprises 40% by weight of green tea (or green tea extract), 20% by weight of Centella asiatica (or Centella asiatica extract), 30% by weight of mugwort (or mugwort extract), 10% by weight of ginger (or ginger extract) may contain
본 발명의 조성물에 포함되는 녹차(또는 녹차 추출물), 병풀(또는 병풀 추출물), 쑥(또는 쑥 추출물), 생강(또는 생강 추출물)과 같은 식물 성분이 상기와 같은 배합비를 벗어나는 경우 독성이 유발되거나 항산화 효과, 미백 효과, 주름 개선 효과, 항염증 효과, 피부 상처의 예방, 개선 또는 치료효과, 보습 효과가 저하될 수 있다. If the plant components such as green tea (or green tea extract), centella asiatica (or centella asiatica extract), mugwort (or mugwort extract), ginger (or ginger extract) contained in the composition of the present invention are out of the above-mentioned mixing ratio, toxicity is induced or The antioxidant effect, the whitening effect, the wrinkle improvement effect, the anti-inflammatory effect, the prevention, improvement or treatment effect of skin wounds, and the moisturizing effect may be reduced.
본 발명에 따른 녹차추출물, 병풀추출물, 쑥추출물, 생강 추출물을 함유하는 입욕제 조성물은 녹차, 병풀, 쑥, 생강을 극성용매로 추출하여 추출물을 제조하는 단계를 포함하여 제조될 수 있다.The bath composition containing green tea extract, centella asiatica extract, mugwort extract, and ginger extract according to the present invention can be prepared by extracting green tea, centella asiatica, mugwort, and ginger with a polar solvent to prepare an extract.
본 발명의 바람직한 구현예에 따르면, 녹차, 병풀, 쑥, 생강 분말을 물, 탄소수 1 내지 6의 알코올 또는 알코올 수용액을 이용하여 추출하여 녹차, 병풀, 쑥, 생강 복합 추출물을 제조할 수 있다.According to a preferred embodiment of the present invention, green tea, centella asiatica, mugwort, and ginger powder may be extracted using water, an alcohol having 1 to 6 carbon atoms, or an aqueous alcohol solution to prepare a green tea, centella asiatica, mugwort, and ginger complex extract.
본 발명의 바람직한 구현예에 따르면, 녹차 추출물, 병풀 추출물, 쑥 추출물, 생강 추출물은 가장 바람직하게는 녹차, 병풀, 쑥, 생강 분쇄물을 이용하여 극성용매로 추출하여 얻어진 추출물이 바람직하게 이용될 수 있다.According to a preferred embodiment of the present invention, green tea extract, centella asiatica extract, mugwort extract, and ginger extract are most preferably extracts obtained by extracting green tea, centella asiatica, mugwort, and ginger pulverized product with a polar solvent. have.
상기 추출은 40-100℃에서 수행되는 것이 바람직한데, 더욱 바람직하게는 30-40℃로 수행해야 한다. 온도가 너무 낮을 경우 효능이 약하게 나타날 수 있고, 추출 시간이 길어지며, 너무 높은 온도에서 추출할 경우 녹차 추출물, 병풀 추출물, 쑥 추출물, 생강 추출물 성분들의 변성이 있을 수 있다.The extraction is preferably performed at 40-100° C., more preferably at 30-40° C. If the temperature is too low, the efficacy may appear weak, the extraction time will be long, and if the extraction is at too high temperature, there may be denaturation of the components of green tea extract, centella asiatica extract, mugwort extract, and ginger extract.
본 발명의 주름 개선 및 항염증기능의 입욕제 조성물은 피부 손상 예방 또는 회복용 식품 조성물과 피부 손상 예방 및 회복용 화장료 조성물의 제조에 이용될 수 있다.The wrinkle-improving and anti-inflammatory bath composition of the present invention can be used to prepare a food composition for preventing or recovering skin damage and a cosmetic composition for preventing and recovering skin damage.
본 발명은 녹차 추출물, 병풀 추출물, 쑥 추출물, 생강 추출물은 통상의 다양한 식품용 성분과 혼합하여 식품을 제조할 수 있다.In the present invention, green tea extract, centella asiatica extract, mugwort extract, and ginger extract can be mixed with various common food ingredients to prepare food.
본 발명의 녹차 추출물, 병풀 추출물, 쑥 추출물, 생강 추출물을 함유하는 화장료 조성물은 화장수류, 에센스류, 크림류, 팩류, 파운데이션류, 메이크업 베이스류 등과 같은 기초제품, 색조제품 및 두발제품, 입욕제 등 다양한 제형으로 제조될 수 있다. 구체적으로 액상, 크림상, 페이스트상 및 고체상 등 다양한 성상으로 적용이 가능하며, 통상적인 화장료 제조법에 적용시킬 수 있다.The cosmetic composition containing green tea extract, centella asiatica extract, mugwort extract, and ginger extract of the present invention can be used in a variety of basic products such as lotions, essences, creams, packs, foundations, makeup bases, color products, hair products, bath products, etc. It can be prepared in a dosage form. Specifically, it can be applied in various properties such as liquid, cream, paste, and solid, and can be applied to a conventional cosmetic preparation method.
본 발명에 따른 녹차, 병풀, 쑥, 생강 분말의 배합비를 통하여 엘라스타아제를 억제하여 주름 개선 효능을 확인하였으며, 멜라닌 형성에 관여하는 타이로시나아제의 활성을 억제하여 미백효과를 확인하였고, 히알루로니다아제의 활성 억제를 통하여 보습효과가 있는 것을 확인하였다. 또한 마크로파아지(macrophage) 세포에서는 항염효과를 확인하고, 각질세포에서 상처치료가 우수하다는 것을 확인되었다. The anti-wrinkle effect was confirmed by inhibiting elastase through the mixing ratio of green tea, centella asiatica, mugwort, and ginger powder according to the present invention, and the whitening effect was confirmed by inhibiting the activity of tyrosinase involved in melanin formation, hyaluronic acid It was confirmed that there was a moisturizing effect by inhibiting the activity of Ronidase. In addition, the anti-inflammatory effect was confirmed in macrophage cells, and it was confirmed that wound healing was excellent in keratinocytes.
따라서 본 발명에 따른 녹차 추출물, 병풀 추출물, 쑥 추출물, 생강 추출물은 엘라스타아제, 타이로시나아제, 및 히알루로니다아제 활성을 억제하고, 마크로파아지 세포에서 항염 효과와 각질세포에서 상처치료를 확인하여 피부질환의 예방과 개선 효과를 나타낸다.Therefore, the green tea extract, centella asiatica extract, mugwort extract, and ginger extract according to the present invention inhibited elastase, tyrosinase, and hyaluronidase activity, and confirmed the anti-inflammatory effect in macrophage cells and wound healing in keratinocytes. Thus, it has the effect of preventing and improving skin diseases.
이하, 본 발명을 구체적으로 설명하기 위해 실시예, 실험예 등의 구체적인 예를 들어 설명하기로 한다. Hereinafter, specific examples such as Examples, Experimental Examples, etc. will be described to describe the present invention in detail.
그러나 본 발명에 따른 실시예 또는 실험예들은 여러 가지 다른 형태로 변형될 수 있으며, 본 발명의 범위가 아래에서 상술하는 예들에 한정되는 것으로 해석되는 것은 아니다.However, the embodiments or experimental examples according to the present invention may be modified in various other forms, and the scope of the present invention is not to be construed as being limited to the examples described below.
실시예 1 : 본 발명에 따른 조성물의 제조 및 추출물의 제조Example 1: Preparation of a composition according to the present invention and preparation of an extract
녹차, 병풀, 쑥, 및 생강을 정제수를 이용하여 이물질이 없도록 세척기로 수회 세척하고, 건조기를 이용하여 건조하였다. 건조는 식물을 쉽게 파쇄할 수 있을 정도로 건조하였다. 건조된 식물은 파쇄기를 이용하여 파쇄하고 녹차 40 중량%, 병풀 20 중량%, 쑥 30 중량%, 생강 10 중량%을 균일하게 혼합하여 조성물을 제조하였다. 상기 혼합된 식물에 정제수 10배를 넣고 40℃로 맞추고 4시간 동안 추출하고 여과하였다. 상기 추출물을 정제수로 100 mg/ml로 용해시켜 스톡 용액을 제조하여 실험에 이용하였다.Green tea, centella asiatica, mugwort, and ginger were washed several times with a washing machine so that there were no foreign substances using purified water, and then dried using a dryer. Drying was dry enough to break the plants easily. The dried plants were crushed using a crusher, and a composition was prepared by uniformly mixing 40% by weight of green tea, 20% by weight of Centella asiatica, 30% by weight of mugwort, and 10% by weight of ginger. 10 times purified water was added to the mixed plant, adjusted to 40° C., extracted for 4 hours, and filtered. The extract was dissolved in purified water at 100 mg/ml to prepare a stock solution and used in the experiment.
실시예 2 : 본 발명에 따른 추출물의 제조Example 2: Preparation of extract according to the present invention
녹차, 병풀, 쑥, 생강을 각각 또는 표 1과 같이 혼합한 식물 50 g에 정제수 500 ml을 가하여 40℃에서 4시간 동안 가열추출하고 여과하였다. 상기 추출물을 정제수로 100 mg/ml로 용해시켜 스톡 용액을 제조하여 실험에 이용하였다.500 ml of purified water was added to 50 g of green tea, centella asiatica, mugwort, and ginger each or a mixed plant as shown in Table 1, extracted by heating at 40° C. for 4 hours, and filtered. The extract was dissolved in purified water at 100 mg/ml to prepare a stock solution and used in the experiment.
실험예 1. 본 발명에 따른 조성물의 항산화 효과 평가Experimental Example 1. Evaluation of the antioxidant effect of the composition according to the present invention
본 발명에 따른 조성물의 항산화 효과를 알아보기 위하여 자유라디칼 소거 실험(Free Radical Scavenging Activity Test)을 실시하였다. 자유라디칼 소거 시험은 Kim 등(Kor. J. Pharmacogn., 24 (4), 299~303(1993))의 방법을 변형한 것으로써, 안정한 자유라디칼인 DPPH(1,1-Diphenyl-2-picrylhydrazyl radical (Sigma)을 사용하였다. In order to examine the antioxidant effect of the composition according to the present invention, a Free Radical Scavenging Activity Test was performed. The free radical scavenging test is a modification of the method of Kim et al. (Kor. J. Pharmacogn., 24 (4), 299~303 (1993)), and the stable free radical DPPH (1,1-Diphenyl-2-picrylhydrazyl radical (Sigma) was used.
먼저, 0.2 mM DPPH 용액 1 ml에 상기 실시예 2에 따라 수득된 식물 추출물 시료를 메탄올에 적당한 농도로 희석하여 2 ml 혼합하고, 실온에서 10분간 방치한 후 517 nm에서 흡광도를 측정한다. First, the plant extract sample obtained according to Example 2 was diluted to an appropriate concentration in methanol in 1 ml of 0.2 mM DPPH solution, mixed with 2 ml, and left at room temperature for 10 minutes, and then absorbance at 517 nm was measured.
한편, 상기 자유라디칼 소거 시험에서의 대조군은 시료용액 대신 메탄올을 넣어 같은 방법으로 측정하며, DPPH 용액 대신 메탄올을 넣어 식물 추출물 시료와 대조군에 대한 각각의 색 보정값을 얻는 것으로 설정하였다. On the other hand, the control in the free radical scavenging test was measured in the same way by putting methanol instead of the sample solution, and adding methanol instead of the DPPH solution to obtain each color correction value for the plant extract sample and the control.
도 1에서 확인할 수 있듯이, 본 발명에 따른 복합 추출물에서는 4:2:3:1, 3:3:3:1, 4:2:2:2 추출물에서 우수한 효과가 나타났다.As can be seen in FIG. 1 , in the complex extract according to the present invention, excellent effects were observed in the 4:2:3:1, 3:3:3:1, and 4:2:2:2 extracts.
실험예 2 Experimental Example 2 : : 본 발명에 따른 조성물의 of the composition according to the invention In vitroin vitro 타이로시나제 활성 저해시험 Tyrosinase activity inhibition test
타이로시나제는 생체 내에서 타이로신(tyrosine)이라는 물질의 산화과정을 촉진하여 멜라닌이 생성되게 도와주는 효소이다. 본 실험예에서는 이 효소의 기능을 억제하여 타이로신의 산화로부터 멜라닌이 형성되는 것을 억제하는 정도를 측정하는 방법(Pomerantz S.H., J. Biochem., 24:161-168, 1966)을 응용해 전술한 실시예 2에 따라 수득된 식물 추출물의 미백효과를 평가하였다.Tyrosinase is an enzyme that helps to produce melanin by accelerating the oxidation process of a substance called tyrosine in a living body. In this experimental example, the above-described implementation by applying the method (Pomerantz S.H., J. Biochem., 24:161-168, 1966) for measuring the degree of inhibiting the formation of melanin from the oxidation of tyrosine by inhibiting the function of this enzyme The whitening effect of the plant extract obtained according to Example 2 was evaluated.
상기 타이로시나제 활성저해시험에 있어 대조군은 식물 추출물 시료 대신 0.1 M 인산완충액(pH 6.8)을 넣어 같은 방법으로 측정하였다. In the tyrosinase activity inhibition test, the control group added 0.1 M phosphate buffer (pH 6.8) instead of the plant extract sample and measured in the same manner.
그 결과는 Significant results according to students test (p < 0.05), (p < 0.01)로서, 도 2에 나타내었다. The results were shown in FIG. 2 as Significant results according to students test (p < 0.05), (p < 0.01).
실험결과, 상기 실시예 2에 따른 식물 추출물의 타이로시나아제활성 저해 효과는 4:2:3:1과 4:2:2:2로 배합된 복합추출물에서 우수한 효과가 나타났다.As a result of the experiment, the tyrosinase activity inhibitory effect of the plant extract according to Example 2 was excellent in the complex extract formulated in 4:2:3:1 and 4:2:2:2.
실험예 3Experimental Example 3 : : 본 발명에 따른 조성물의 엘라스테이즈 활성저해 시험Elastase activity inhibition test of the composition according to the present invention
상기 각 실시예 2에 따라 수득된 식물 추출물을 시료로 하여 피부의 주름 억제 및 개선효과 검정시험으로서 엘라스테이즈 활성 저해 시험(Elastase Inhibition Activity Test)을 실시하였다.An Elastase Inhibition Activity Test was performed as a test for skin wrinkle suppression and improvement by using the plant extract obtained according to Example 2 as a sample.
엘라스틴(Elastin)을 분해하는 효소인 엘라스테이즈의 활성을 측정하는 방법으로 엘라스테이즈 기질인 N-Succinyl-Ala-Ala-Ala ρ-nitroaniline을 이용하여 ρ-nitroaniline이 분해되면서 생기는 색의 변화를 410 nm의 파장에서 흡광도를 측정함으로써 엘라스테이즈 활성을 측정하는 방법이다. 완충액은 pH 8.0, 0.267 M Trizma-HCl (Sigma), 기질액은 8.8 mM N-Succinyl-(Ala)3 ρ-nitroaniline (Sigma), 효소액은 돼지췌장 엘라스테이즈를 10 ㎍/ml (Sigma)의 농도로 사용하였다. 완충액 60 μl, 기질액 20 μl와 상기 각 실시예 1, 2에 따라 수득된 각 식물 추출물 각각을 농도별로 정제수에 녹여 100 μl로 한 시료액을 섞은 후, 효소액 20 μl를 넣어 25℃ 항온수조에서 15분간 반응시켜 ρ-nitroaniline의 생성량을 마이크로 플레이트 판독기를 이용하여 파장 405 nm에서 측정한다.As a method of measuring the activity of elastase, an enzyme that decomposes elastin, the color change caused by the decomposition of ρ-nitroaniline is measured using N-Succinyl-Ala-Ala-Ala ρ-nitroaniline, which is an elastase substrate. This is a method for measuring elastase activity by measuring absorbance at a wavelength of 410 nm. The buffer solution is pH 8.0, 0.267 M Trizma-HCl (Sigma), the substrate solution is 8.8 mM N-Succinyl-(Ala) 3 ρ-nitroaniline (Sigma), and the enzyme solution is 10 μg/ml (Sigma) of pig pancreatic elastase. concentration was used. 60 μl of the buffer solution, 20 μl of the substrate solution, and each of the plant extracts obtained according to Examples 1 and 2 above were dissolved in purified water at each concentration to make 100 μl of the sample solution, and then 20 μl of the enzyme solution was added in a water bath at 25° C. After reacting for 15 minutes, the amount of ρ-nitroaniline produced is measured at a wavelength of 405 nm using a microplate reader.
한편, 상기 엘라스테이즈 활성도를 측정하기 위한 대조군은 식물 추출물 대신 정제수를 넣어 같은 방법으로 측정하였으며, 효소액 대신 정제수를 넣어 각각에 대한 색 보정값을 얻은 경우를 설정하였다(참고문헌: Lim, Suk-Won외, The effect of two terpenoids, ursolic acid and oleanolic acid on epidermal permeability barrier and simultaneously on dermal functions, 대한화장품학회지, 29(2), 205-232, 2003).On the other hand, the control group for measuring the elastase activity was measured in the same manner by adding purified water instead of a plant extract, and a case where purified water was added instead of an enzyme solution to obtain color correction values for each was set (References: Lim, Suk- Won et al., The effect of two terpenoids, ursolic acid and oleanolic acid on epidermal permeability barrier and simultaneously on dermal functions, Journal of the Korean Cosmetic Society, 29(2), 205-232, 2003).
그 결과는 Significant results according to students test (p < 0.05), (p < 0.01)로서, 도 3에 나타내었다. The results were shown in FIG. 3 as Significant results according to students test (p < 0.05), (p < 0.01).
실험결과, 상기 실시예 2에 따른 식물 추출물의 엘라스테이즈 활성 저해 효과는 4:2:3:1과 4:2:2:2로 배합된 복합추출물에서 우수한 효과를 보였다.As a result of the experiment, the elastase activity inhibitory effect of the plant extract according to Example 2 was excellent in the complex extracts formulated in 4:2:3:1 and 4:2:2:2.
실험예 4 : 본 발명에 따른 조성물의 항염증 시험Experimental Example 4: Anti-inflammatory test of the composition according to the present invention
상기 실시예 2에서 얻어진 식물 추출물의 항염 활성 효과를 확인해 보기 위해 염증 유발에 중요한 역할을 하는 것으로 알려진 NO (nitric oxide) 생성에 대한 효과를 알아보았다. 일반적인 NO의 형성은 박테리아를 죽이거나 종양을 제거시키는 중요한 역할을 하지만, 염증상태에서 iNOS에 의해 생성된 NO는 혈관 투과성, 부종 등의 염증반응을 촉진시킬 뿐만 아니라 염증매개체의 생합성을 촉진하여 염증을 심화시키는 것으로 알려져 있다.In order to confirm the anti-inflammatory activity of the plant extract obtained in Example 2, the effect on NO (nitric oxide) production, which is known to play an important role in inducing inflammation, was investigated. Although the formation of general NO plays an important role in killing bacteria or removing tumors, NO produced by iNOS in an inflammatory state not only promotes inflammatory reactions such as vascular permeability and edema, but also promotes biosynthesis of inflammatory mediators to reduce inflammation. known to exacerbate it.
RAW264.7 세포를 10% FBS가 첨가된 DMEM 배지를 이용하여 5×105 cells/well로 조절한 후 24 웰 플레이트에 접종하고, 시료를 처리하고 30분 배양 후 LPS (lipopolysaccharide, 1 μg/ml)를 첨가하여 24시간 배양하였다. 생성된 NO의 양은 NO detection kit (iNtRON)를 이용하여 측정한 값을 나타내었다. 상기 항염 활성증 효과 시험을 위한 대조군은 LPS를 단독으로 처리한 실험군으로 하였다.RAW264.7 cells were adjusted to 5×10 5 cells/well using DMEM medium supplemented with 10% FBS, and then inoculated in a 24-well plate, sampled and incubated for 30 minutes, LPS (lipopolysaccharide, 1 μg/ml ) was added and cultured for 24 hours. The amount of NO generated was measured using a NO detection kit (iNtRON). The control group for the anti-inflammatory activity test was the experimental group treated with LPS alone.
그 결과는 Significant results according to students test (p < 0.05), (p < 0.01) 로서, 도 4에 나타내었다. The results were shown in FIG. 4 as Significant results according to students test (p < 0.05), (p < 0.01).
도 4의 측정결과를 살펴보면, 4:3:2:1과 4:2:3:1로 배합된 복합추출물에서 NO의 감소 효과가 우수한 것으로 나타났다.Looking at the measurement results in FIG. 4 , it was found that the NO reduction effect was excellent in the complex extracts formulated in 4:3:2:1 and 4:2:3:1.
실험예 5 : 본 발명에 따른 조성물의 육안적 형태 분석을 통한 상처의 개선 또는 치료 효과 검정Experimental Example 5: Test of wound improvement or therapeutic effect through gross morphological analysis of the composition according to the present invention
세포배양용기 웰 당 2.5 x 105 농도로 준비된 HaCaT 세포를 분주한 다음 248시간 배양하여 세포 단일층을 형성시킨 후 배양배지를 제거한 다음 p200 피펫팁으로 “긁힘-손상”을 유도하였다. “긁힘-손상”이 유도된 상기 세포에 상기 실시예 2에서 얻어진 식물 추출물을 포함한 배양 배지를 세포에 첨가한 다음 48시간 동안 배양했다.After dispensing the prepared HaCaT cells at a concentration of 2.5 x 10 5 per well of the cell culture vessel and culturing for 248 hours to form a cell monolayer, the culture medium was removed and “scratch-damage” was induced with a p200 pipette tip. A culture medium containing the plant extract obtained in Example 2 was added to the cells induced in "scratches-damage", and then cultured for 48 hours.
본 발명에서는 시험관 내 상처치유 측정법(in vitro woound healing assay)을 이용하여 “긁힘-손상”세포층에 추출물을 50 μg/ml 농도로 48시간 각각 처리한 후 긁힘 상처가 회복되는 정도를 확인하였다. 대조군은 추출물을 녹인 DMSO를 동일 함량 처리하였다.In the present invention, the degree of recovery of the scratch wound was confirmed after each treatment of the extract at a concentration of 50 μg/ml in the “scratch-damaged” cell layer for 48 hours using an in vitro woound healing assay. The control group was treated with the same amount of DMSO in which the extract was dissolved.
실험결과, 복합물 4:3:2:1, 4:2:3:1, 3:3:3:1에서는 긁힌 면적이 거의 감소되지 않았으며, 복합물 4:2:2:2의 50 μg/ml에서는 “긁힘-손상”이 48시간 후 긁힌 면적이 감소되었음을 확인 할 수 있다.As a result of the experiment, the scratched area was hardly reduced in the composites 4:3:2:1, 4:2:3:1, 3:3:3:1, and 50 μg/ml of the composite 4:2:2:2 In “scratch-damage”, it can be confirmed that the scratched area is reduced after 48 hours.
실험예 6 : 본 발명에 따른 조성물의 보습 효과 검정Experimental Example 6: Moisturizing effect test of the composition according to the present invention
상기 각 실시예 2에 따라 수득된 식물 추출물을 시료로 하여 피부의 보습 효과 검정시험으로서 히알루로니다아제 활성 저해 시험(Hyaluronidase Inhibition Activity Test)을 실시하였다.A Hyaluronidase Inhibition Activity Test was performed as a test for moisturizing the skin using the plant extract obtained according to Example 2 as a sample.
히알루로니다아제 효소의 활성억제 실험은 다음과 같이 진행하였다. 0.1M acetate buffer (pH 3.5)에 녹인 히알루로니다아제(7,900 unit/ml) 0.05 ml와 시료용액 0.1 ml을 혼합하여 37℃에서 20분간 반응 시킨 다음 12.5 mM 염화칼슘(CaCl2) 0.1 ml을 가하고 혼합 후 다시 20분간 반응시켰다. 대조군은 히알루로니다아제(7,900 unit/ml) 0.05 ml과 추출물 대신 증류수 0.1 ml을 혼합하고, 기질로서 0.1M acetate buffer (pH 3.5)에 녹인 히아루론산(hyaluronic, HA; 12 mg/ml)을 첨가하여 다시 40분간 반응시켜 0.4 N potassium tetraborate 0.1 ml 및 0.4 N NaOH 용액을 0.1 ml 반응 혼합물에 첨가하여 3분 동안 수욕상에서 가열한 후 완전히 냉각시켰다. 냉각시킨 반응물에 발색제인 p-dimethylamino benzaldehyde (DMAB) 시약 3 ml를 가하여 37℃에서 20분간 반응시킨 다음 585 nm에서 흡광도를 측정하였다.The activity inhibition experiment of the hyaluronidase enzyme was performed as follows. 0.05 ml of hyaluronidase (7,900 unit/ml) dissolved in 0.1M acetate buffer (pH 3.5) and 0.1 ml of the sample solution were mixed and reacted at 37°C for 20 minutes, then 0.1 ml of 12.5 mM calcium chloride (CaCl 2 ) was added and mixed. Then, it was reacted again for 20 minutes. As a control group, 0.05 ml of hyaluronidase (7,900 unit/ml) and 0.1 ml of distilled water were mixed instead of the extract, and hyaluronic acid (hyaluronic, HA; 12 mg/ml) dissolved in 0.1M acetate buffer (pH 3.5) as a substrate was added. After reacting for another 40 minutes, 0.1 ml of 0.4 N potassium tetraborate and 0.4 N NaOH solution were added to the 0.1 ml reaction mixture, heated in a water bath for 3 minutes, and then completely cooled. 3 ml of p-dimethylamino benzaldehyde (DMAB) reagent, a color developer, was added to the cooled reaction mixture, reacted at 37° C. for 20 minutes, and absorbance was measured at 585 nm.
실험결과, 복합물 4:3:2:1, 4:2:3:1, 3:3:3:1, 4:2:2:2 에서는 대조군 대비 히알루로니다아제 활성이 억제되는 것을 확인하였으며 특히, 복합물 4:2:2:2에서 가장 우수한 효과를 확인하였다.As a result of the experiment, it was confirmed that the hyaluronidase activity was inhibited in the complexes 4:3:2:1, 4:2:3:1, 3:3:3:1, 4:2:2:2 compared to the control group. , the most excellent effect was confirmed in the complex 4:2:2:2.
40℃의 낮은 온도에서 추출한 추출물의 효능 결과인 도 1, 도 2, 도 3, 도 4, 도 5 및 도6에서의 결과를 보면 복합추출물에서는 4:2:3:1, 4:3:2:1, 4:2:2:2 추출물에서는 항산화, 미백, 주름개선, 항염 및 보습 효과가 나타났으며 특히 4:2:3:1에서는 항산화, 미백, 주름개선, 항염에서 더 우수한 효과를 보였다. 그러나 상처 개선 및 보습 효과는 4:2:2:2에서 가장 좋은 효과를 보였다.Looking at the results in FIGS. 1, 2, 3, 4, 5 and 6, which are the efficacy results of the extract extracted at a low temperature of 40°C, in the complex extract, 4:2:3:1, 4:3:2 :1, 4:2:2:2 extract showed antioxidant, whitening, anti-wrinkle, anti-inflammatory and moisturizing effects, and 4:2:3:1 showed better antioxidant, whitening, anti-wrinkle and anti-inflammatory effects. . However, the wound improvement and moisturizing effect showed the best effect at 4:2:2:2.
본 발명에 따르면, 녹차, 병풀, 쑥, 생강 혼합 분말을 유효성분으로 함유한 입욕제 조성물은 항산화, 미백, 주름개선, 상처치유 및 보습 효과가 우수하며, 독성도 없는 것이므로 천연 바이오 소재로 화장료나 식품용 소재로 널리 활용이 가능하다.According to the present invention, the bath composition containing green tea, centella asiatica, mugwort, and ginger mixed powder as an active ingredient is excellent in antioxidant, whitening, wrinkle improvement, wound healing and moisturizing effects, and is non-toxic, so it is a natural bio material for cosmetics or food. It can be widely used as a material for
또한, 뛰어난 천연 바이오 소재 조성물을 제공하기 때문에 생명바이오 식품·의약 산업상 매우 유용한 것이다.In addition, since it provides an excellent natural biomaterial composition, it is very useful in the bio-bio food/pharmaceutical industry.
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