KR102271861B1 - Food composition for skin improvement with natural material extracts - Google Patents
Food composition for skin improvement with natural material extracts Download PDFInfo
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- KR102271861B1 KR102271861B1 KR1020200159140A KR20200159140A KR102271861B1 KR 102271861 B1 KR102271861 B1 KR 102271861B1 KR 1020200159140 A KR1020200159140 A KR 1020200159140A KR 20200159140 A KR20200159140 A KR 20200159140A KR 102271861 B1 KR102271861 B1 KR 102271861B1
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- KR
- South Korea
- Prior art keywords
- extract
- bellflower
- pomegranate
- pear
- skin
- Prior art date
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Images
Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- 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
- A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
-
- 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
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- 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
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/318—Foods, ingredients or supplements having a functional effect on health having an effect on skin health and hair or coat
-
- 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
- A23V2250/00—Food ingredients
- A23V2250/20—Natural extracts
- A23V2250/21—Plant extracts
-
- 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
- A23V2300/00—Processes
- A23V2300/14—Extraction
Abstract
Description
본 발명은 먹는 화장품으로서 천연소재추출물을 포함하며 피부 주름 개선 또는 완화, 피부 미백, 피부 유수분 균형 유지 및 피부 보습 효과를 발휘하는 식품 조성물에 관한 것이다.The present invention relates to a food composition that contains a natural material extract as an edible cosmetic and exhibits skin wrinkle improvement or alleviation, skin whitening, skin oil-water balance maintenance and skin moisturizing effect.
현대사회는 고령화라는 시대적 흐름과 함께 웰빙 라이프, 셀프 메디케이션 트렌드가 확산되고 있으며, 이에 따른 건강기능식품 시장이 활발하게 확대되고 있다. 자연스럽고 건강한 피부에 대한 관심이 높아지면서 이너뷰티(inner beauty), 즉 먹는 화장품에 대한 관심도 증가하고 있다.In modern society, well-being lifestyle and self-medication trends are spreading along with the trend of aging, and the health functional food market is actively expanding accordingly. As interest in natural and healthy skin increases, interest in inner beauty, that is, edible cosmetics, is also increasing.
피부 주름 개선, 피부 미백, 피부 보습 효과를 위해 일반적으로는 화장품이나 의약품을 피부에 직접 도포하여 사용한다. 그러나 이러한 피부 외용제는 효과가 일시적이고, 또한 도포 부위에만 국소적인 효과를 주는 한계가 있다. 반면 기능성 물질이 포함된 이너뷰티(inner beauty) 소재는 피부에 직접 도포되는 것이 아니라 경구 섭취에 따라 표피층 전체와 진피층의 피부 조직에 작용하여 주름 개선, 미백, 보습 효과를 나타낼 수 있다.For skin wrinkle improvement, skin whitening, and skin moisturizing effects, cosmetics or pharmaceuticals are generally applied directly to the skin and used. However, these external preparations for the skin have a temporary effect, and also have limitations in giving a local effect only to the application site. On the other hand, inner beauty materials containing functional substances are not applied directly to the skin, but act on the entire epidermal layer and the skin tissue of the dermis layer according to oral intake, thereby improving wrinkles, whitening, and moisturizing effects.
이너뷰티(inner beauty) 소재들은 대부분 외국에서 연구된 결과와 생산된 원료를 이용하여 제품을 개발 또는 생산하고 있다. 이에 국내 이너뷰티(inner beauty) 시장의 성장을 위해서는 국내 소재의 피부 개선 효과에 대한 연구 개발이 지속적으로 필요한 실정이다.Inner beauty materials are mostly developed or produced using the results of research in foreign countries and raw materials produced. Accordingly, for the growth of the domestic inner beauty market, research and development on the skin improvement effect of domestic materials is continuously required.
본 발명은 도라지, 배, 석류, 자소엽에 포함된 폴리페놀을 최대 효율로 추출함에 따라, 각 소재의 유효성분이 최대로 포함되는 조성물을 제공하고자 한다.The present invention seeks to provide a composition in which the active ingredient of each material is maximally extracted as polyphenols contained in bellflower, pear, pomegranate, and perilla leaf are extracted with maximum efficiency.
본 발명은 도라지추출물, 배추출물, 석류추출물, 자소엽추출물을 포함함에 따라 피부 주름 개선 또는 완화, 피부 미백, 피부 유수분 균형 유지 및 피부 보습 효과를 발휘하는 식품 조성물을 제공하고자 한다.The present invention intends to provide a food composition that contains bellflower extract, pear extract, pomegranate extract, and perilla leaf extract, thereby improving or alleviating skin wrinkles, skin whitening, maintaining skin oil-water balance, and moisturizing skin.
본 발명은 기존의 피부 외용제가 도포 부위에만 국소적으로 작용하는 한계를 극복하고자 한다.An object of the present invention is to overcome the limitation that the existing external preparation for skin acts only locally on the application site.
본 발명은 도라지추출물, 배추출물, 석류추출물 및 자소엽추출물을 포함하는 피부 주름 개선용 식품 조성물을 제공한다.The present invention provides a food composition for improving skin wrinkles comprising a bellflower extract, a pear extract, a pomegranate extract and a perilla leaf extract.
또한, 본 발명은 도라지추출물, 배추출물, 석류추출물 및 자소엽추출물을 포함하는 피부 미백용 식품 조성물을 제공한다.In addition, the present invention provides a food composition for skin whitening comprising a bellflower extract, a pear extract, a pomegranate extract and a perilla leaf extract.
본 발명은 도라지추출물, 배추출물, 석류추출물 및 자소엽추출물을 포함하는 피부 보습용 식품 조성물을 제공한다.The present invention provides a skin moisturizing food composition comprising a bellflower extract, a pear extract, a pomegranate extract and a perilla leaf extract.
본 발명에 있어서, 상기 도라지추출물, 배추출물, 석류추출물 및 자소엽추출물은, 바람직하게는 추출용매로 정제수를 이용한 것이 좋다.In the present invention, the bellflower extract, pear extract, pomegranate extract and perilla leaf extract are preferably purified water as an extraction solvent.
본 발명에 있어서, 상기 도라지추출물은, 바람직하게는 51~55 ℃에서 25~29 시간 추출한 것이 좋다.In the present invention, the bellflower extract is preferably extracted at 51-55 ° C. for 25-29 hours.
본 발명에 있어서, 상기 배추출물은, 바람직하게는 94~98 ℃에서 18~22 시간 추출한 것이 좋다.In the present invention, the pear extract is preferably extracted at 94-98 ° C. for 18-22 hours.
본 발명에 있어서, 상기 석류추출물은, 바람직하게는 94~98 ℃에서 12~16 시간 추출한 것이 좋다.In the present invention, the pomegranate extract is preferably extracted at 94 to 98° C. for 12 to 16 hours.
본 발명에 있어서, 상기 자소엽추출물은, 바람직하게는 51~55 ℃에서 25~29 시간 추출한 것이 좋다.In the present invention, the perilla leaf extract is preferably extracted at 51-55 ° C. for 25-29 hours.
본 발명은 도라지추출물, 배추출물, 석류추출물, 자소엽추출물에 대하여 폴리페놀을 추출할 수 있는 최적의 추출조건을 도출하고, 이들을 이용하여 피부 주름 개선 또는 완화, 피부 미백, 피부 유수분 균형 유지 및 피부 보습 효과를 발휘하는 식품 조성물을 제공한다.The present invention derives optimal extraction conditions for extracting polyphenols from bellflower extract, pear extract, pomegranate extract, and perilla leaf extract, and uses them to improve or alleviate skin wrinkles, skin whitening, skin oil-water balance maintenance and skin A food composition exhibiting a moisturizing effect is provided.
본 발명은 기존의 피부 외용제가 가지는 한계를 극복하고, 경구 섭취에 따라 표피층 전체와 진피층의 피부 조직에 피부 개선 효과가 전달되는 먹는 화장품을 제공한다.The present invention overcomes the limitations of the existing external preparation for skin and provides an edible cosmetic in which the skin improvement effect is delivered to the entire epidermal layer and the skin tissue of the dermal layer according to oral intake.
도 1은 도라지의 폴리페놀 추출 조건 확립을 위한 반응표면분석법에 따른 등고선도와 표면도이다.
도 2는 도라지의 폴리페놀 추출 조건 확립을 위한 반응표면분석법의 결과를 나타낸 도면이다.
도 3은 배의 폴리페놀 추출 조건 확립을 위한 반응표면분석법에 따른 등고선도와 표면도이다.
도 4는 배의 폴리페놀 추출 조건 확립을 위한 반응표면분석법의 결과를 나타낸 도면이다.
도 5은 석류의 폴리페놀 추출 조건 확립을 위한 반응표면분석법에 따른 등고선도와 표면도이다.
도 6는 석류의 폴리페놀 추출 조건 확립을 위한 반응표면분석법의 결과를 나타낸 도면이다.
도 7은 자소엽의 폴리페놀 추출 조건 확립을 위한 반응표면분석법에 따른 등고선도와 표면도이다.
도 8는 자소엽의 폴리페놀 추출 조건 확립을 위한 반응표면분석법의 결과를 나타낸 도면이다.
도 9는 각 추출물의 B16-F10 세포에 대한 세포독성을 나타낸 도면이다.
도 10은 각 추출물의 WI-38 세포에 대한 세포독성을 나타낸 도면이다.
도 11은 각 추출물의 HaCaT 세포에 대한 세포독성을 나타낸 도면이다.
도 12는 각 추출물의 멜라닌 생성 억제 효과를 나타낸 도면이다.
도 13은 각 추출물의 프로콜라겐 생성 효과를 나타낸 도면이다.
도 14는 각 추출물의 히알루론산 생성 효과를 나타낸 도면이다.
도 15는 천연소재복합추출물의 배합비에 따른 멜라닌 생성 억제 효과를 나타낸 도면이다.
도 16는 천연소재복합추출물의 배합비에 따른 프로콜라겐 생성 효과를 나타낸 도면이다.
도 17는 천연소재복합추출물의 배합비에 따른 히알루론산 생성 효과를 나타낸 도면이다.1 is a contour diagram and a surface diagram according to a reaction surface analysis method for establishing polyphenol extraction conditions of bellflower.
2 is a view showing the results of the reaction surface analysis method for establishing the polyphenol extraction conditions of bellflower.
3 is a contour diagram and a surface diagram according to a reaction surface analysis method for establishing polyphenol extraction conditions of pears.
4 is a view showing the results of the reaction surface analysis method for establishing the polyphenol extraction conditions of pears.
5 is a contour diagram and a surface diagram according to a reaction surface analysis method for establishing polyphenol extraction conditions of pomegranate.
6 is a view showing the results of the reaction surface analysis method for establishing the polyphenol extraction conditions of pomegranate.
7 is a contour diagram and a surface diagram according to a reaction surface analysis method for establishing polyphenol extraction conditions of perilla leaflets.
8 is a view showing the results of the reaction surface analysis method for establishing the polyphenol extraction conditions of perilla leaf.
9 is a diagram showing the cytotoxicity of each extract to B16-F10 cells.
10 is a diagram showing the cytotoxicity of each extract to WI-38 cells.
11 is a diagram showing the cytotoxicity of each extract to HaCaT cells.
12 is a view showing the melanin production inhibitory effect of each extract.
13 is a diagram showing the procollagen production effect of each extract.
14 is a diagram showing the hyaluronic acid production effect of each extract.
15 is a view showing the effect of inhibiting melanin production according to the mixing ratio of the natural material complex extract.
16 is a view showing the effect of producing procollagen according to the mixing ratio of the natural material complex extract.
17 is a view showing the hyaluronic acid production effect according to the mixing ratio of the natural material complex extract.
본 발명은 이너뷰티(inner beauty), 즉 먹는 화장품에 관한 것으로, 도라지추출물, 배추출물, 석류추출물, 자소엽추출물을 포함하는 식품 조성물을 제공한다. 구체적으로, 본 발명은 도라지추출물, 배추출물, 석류추출물 및 자소엽추출물을 포함하는 피부 주름 개선용 식품 조성물, 피부 미백용 식품 조성물, 피부 보습용 식품 조성물을 제공한다.The present invention relates to inner beauty, ie, cosmetics to be eaten, and provides a food composition comprising a bellflower extract, a pear extract, a pomegranate extract, and a perilla leaf extract. Specifically, the present invention provides a food composition for improving skin wrinkles, a food composition for skin whitening, and a food composition for skin moisturizing, comprising a bellflower extract, a pear extract, a pomegranate extract and a perilla leaf extract.
도라지는 초롱꽃과의 여러해살이풀로, 질경, 길경, 길경채, 도랏, 백약 등으로도 불린다. 도라지는 뿌리를 식용으로 사용하며 빈혈 개선, 편도염 개선, 항암효과 등이 있다고 알려져 있다.Bellflower is a perennial plant of the bellflower family, and is also called plantain, gilgyeong, gilgyeongchae, dorat, and baekyak. Bellflower roots are used for food and are known to have anti-anemia, tonsillitis, and anti-cancer effects.
배는 장미과에 속하는 배나무의 열매로 껍질은 노란색, 그을린 갈색을 띠며 속살은 오돌토돌한 식감이 느껴진다. 배는 생과일, 음료, 주류 등에 다양하게 활용된다. 배는 이뇨작용 촉진, 변비 예방, 호흡기 질환 예방 효과가 있다고 알려져 있다.Pears are the fruits of the pear tree belonging to the Rosaceae family. The skin is yellow and charred brown, and the flesh has a crunchy texture. Pears are widely used for fresh fruits, beverages, and alcoholic beverages. Pears are known to promote diuresis, prevent constipation, and prevent respiratory diseases.
석류는 둥근 모양의 단단하고 노르스름한 외관이 과육을 감싸고 있는데, 과육에는 다수의 종자가 포함되어 있다. 석류는 과육을 섭취하거나 과일주 등의 음료 형태로 섭취한다. 열매와 껍질 모두 고혈압, 동맥경화 예방 효과가 있다고 알려져 있다.A pomegranate has a round, hard, yellowish exterior that surrounds the flesh, which contains many seeds. Pomegranate is consumed by eating the flesh or in the form of beverages such as fruit juice. Both the fruit and the peel are known to be effective in preventing hypertension and arteriosclerosis.
자소엽은 소엽의 잎이나 끝가지를 말한다. 소엽은 꿀풀과의 한해살이풀로 차조기, 차즈기라고도 한다. 소화장애 해소, 기침 또는 천식 개선 등의 효과가 있다고 알려져 있다.A perilla leaf refers to the leaf or tip of a leaflet. Leaflets are annual plants of the Lamiaceae family, also called perilla and chazugi. It is known to be effective in relieving digestive disorders and improving cough or asthma.
본 발명에 있어서, 도라지추출물, 배추출물, 석류추출물, 자소엽추출물은 당업계에 공지된 통상의 방법에 따라, 즉 통상적인 온도와 압력의 조건 하에서 통상적인 용매를 사용하여 제조될 수 있으며, 바람직하게는 정제수를 이용하는 것이 좋다.In the present invention, bellflower extract, pear extract, pomegranate extract, and perilla leaf extract can be prepared according to a conventional method known in the art, that is, under normal temperature and pressure conditions, using a conventional solvent. It is better to use purified water.
본 발명은 도라지, 배, 석류, 자소엽에 포함된 폴리페놀을 최대 효율로 추출할 수 있는 최적의 추출조건을 확립하였다. 본 발명의 천연소재는 추출용매로 정제수를 사용하였다. 반응표면분석법에 따라 본 발명의 도라지추출물, 배추출물, 석류추출물, 자소엽추출물의 추출조건을 확립하였다.The present invention established optimal extraction conditions for extracting polyphenols contained in bellflower, pear, pomegranate and perilla leaves with maximum efficiency. For the natural material of the present invention, purified water was used as an extraction solvent. The extraction conditions of the bellflower extract, pear extract, pomegranate extract, and perilla leaf extract of the present invention were established according to the response surface analysis method.
본 발명의 도라지추출물은 53.7868 ℃, 27.7279 h 조건으로 추출할 경우 폴리페놀 함량이 최대 29.69 mg/g 추출되는 점을 확인하였다. 즉, 본 발명의 도라지추출물은 바람직하게는 51~55 ℃, 25~29 h 조건으로 추출할 경우 폴리페놀이 효과적으로 추출되는 점을 알 수 있었다.When the bellflower extract of the present invention was extracted under the conditions of 53.7868 °C and 27.7279 h, it was confirmed that the polyphenol content was extracted at a maximum of 29.69 mg/g. That is, it was found that the bellflower extract of the present invention preferably extracts polyphenols effectively at 51-55 ℃ and 25-29 h conditions.
본 발명의 배추출물은 96.2132 ℃, 20.0140 h 조건으로 추출할 경우 폴리페놀 함량이 최대 35.84 mg/g 추출되는 점을 확인하였다. 즉, 본 발명의 배추출물은 바람직하게는 94~98 ℃, 18~22 h 조건으로 추출할 경우 폴리페놀이 효과적으로 추출되는 점을 알 수 있었다.When the pear extract of the present invention was extracted under the conditions of 96.2132 ℃ and 20.0140 h, it was confirmed that the polyphenol content was extracted up to 35.84 mg/g. That is, it was found that the polyphenols were effectively extracted when the pear extract of the present invention was preferably extracted under the conditions of 94 to 98 ℃ and 18 to 22 h.
본 발명의 석류추출물은 96.2132 ℃, 14.1000 h 조건으로 추출할 경우 폴리페놀 함량이 최대 37.31 mg/g 추출되는 점을 확인하였다. 즉, 본 발명의 석류추출물은 바람직하게는 94~98 ℃, 12~16 h 조건으로 추출할 경우 폴리페놀이 효과적으로 추출되는 점을 알 수 있었다.When the pomegranate extract of the present invention was extracted under the conditions of 96.2132 °C and 14.1000 h, it was confirmed that the maximum polyphenol content was extracted at 37.31 mg/g. That is, it was found that polyphenols were effectively extracted when the pomegranate extract of the present invention was preferably extracted under conditions of 94 to 98 °C and 12 to 16 h.
본 발명의 자소엽추출물은 53.7868 ℃, 27.7279 h 조건으로 추출할 경우 폴리페놀 함량이 최대 33.02 mg/g 추출되는 점을 확인하였다. 즉, 본 발명의 자소엽추출물은 바람직하게는 51~55 ℃, 25~29 h 조건으로 추출할 경우 폴리페놀이 효과적으로 추출되는 점을 알 수 있었다.When the perilla leaf extract of the present invention was extracted under the conditions of 53.7868 °C and 27.7279 h, it was confirmed that the maximum polyphenol content was 33.02 mg/g. That is, it was found that the polyphenols were effectively extracted when the perilla leaf extract of the present invention was preferably extracted under the conditions of 51-55° C. and 25-29 h.
한편, 본 발명에서 확립한 각 소재의 추출조건에 따라 제조된 도라지추출물, 배추출물, 석류추출물, 자소엽추출물은 각각 0.1, 0.25, 0.5 및 1 mg/ml 농도로 단독처리 할 경우 세포독성을 나타내지 않는 점을 확인하였다.On the other hand, bellflower extract, pear extract, pomegranate extract, and perilla leaf extract prepared according to the extraction conditions of each material established in the present invention do not show cytotoxicity when treated alone at concentrations of 0.1, 0.25, 0.5 and 1 mg/ml, respectively. It was confirmed that it is not.
또한, 본 발명에서 확립한 각 소재의 추출조건에 따라 제조된 도라지추출물, 배추출물, 석류추출물, 자소엽추출물의 멜라닌 생성 억제능, 프로콜라겐 생성능, 히알루론산 생성능을 확인한 결과, 석류추출물과 자소엽추출물은 멜라닌 생성 억제 효과가 우수하였고, 도라지추출물, 배추출물, 석류추출물은 프로콜라겐 생성 효과가 우수하였고, 도라지추출물, 배추출물, 석류추출물, 자소엽추출물 모두 히알루론산 생성 효과가 우수한 점을 확인하였다.In addition, as a result of confirming the melanin production inhibitory ability, procollagen production capability, and hyaluronic acid production capability of the bellflower extract, pear extract, pomegranate extract, and perilla leaf extract prepared according to the extraction conditions of each material established in the present invention, pomegranate extract and perilla leaf extract was excellent in inhibiting melanin production, bellflower extract, pear extract, and pomegranate extract were excellent in procollagen production, and bellflower extract, pear extract, pomegranate extract, and perilla leaf extract were all excellent in hyaluronic acid production.
한편, 본 발명의 도라지추출물, 배추출물은 멜라닌 생성 억제 효과가 미미하였으며, 자소엽추출물은 프로콜라겐 생성 효과가 미미한 것으로 나타났다. 이에 본 발명은 도라지추출물, 배추출물, 석류추출물, 자소엽추출물을 복합물로 처리할 경우의 멜라닌 생성 억제능, 프로콜라겐 생성능, 히알루론산 생성능을 확인하였다.On the other hand, the bellflower extract and pear extract of the present invention had insignificant melanin production inhibitory effect, and the perilla leaf extract showed insignificant procollagen production effect. Accordingly, the present invention confirmed the ability to inhibit melanin production, procollagen production, and hyaluronic acid production when the bellflower extract, pear extract, pomegranate extract, and perilla leaf extract were treated with a complex.
본 발명의 천연소재복합물은 하기 표 17에 따른 모든 배합비에서 멜라닌 생성 억제 효과를 보였으며, 일부 배합비를 제외하고 프로콜라겐 생성 효과, 히알루론산 생성 효과를 보이는 점을 확인하였다. 즉, 각 추출물을 단독으로 처리할 경우 미미한 피부 개선 효과를 복합물로 처리시에 보완할 수 있는 점을 확인하였다.It was confirmed that the natural material composite of the present invention showed a melanin production inhibitory effect in all the mixing ratios according to Table 17 below, and showed a procollagen production effect and a hyaluronic acid production effect except for some mixing ratios. That is, it was confirmed that when each extract was treated alone, the insignificant skin improvement effect could be supplemented when treated with a complex.
본 발명의 식품 조성물은 피부 미백효과, 피부 주름 개선, 완화 또는 예방 효과, 피부 유수분 균형 유지 및 피부 보습 효과를 발휘한다. 본 발명의 식품 조성물은 경구 투여용 담체를 추가로 포함할 수 있으며, 일 예로 락토스, 전분, 셀룰로스 유도체, 마그네슘 스테아레이트, 스테아르산 등을 포함할 수 있다.The food composition of the present invention exhibits a skin whitening effect, skin wrinkle improvement, alleviation or prevention effect, skin oil-water balance maintenance and skin moisturizing effect. The food composition of the present invention may further include a carrier for oral administration, and may include, for example, lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like.
또한 본 발명의 식품 조성물은 경구투여를 위한 고형제제로 제공될 수 있는데, 일 예로 정제, 환제, 산제, 과립제, 액제, 겔제, 시럽제, 슬러리제, 현탁액 또는 캡슐제 등이 포함되며, 이러한 고형제제는 적어도 하나 이상의 부형제 예를 들면, 전분(옥수수 전분, 밀 전분, 쌀 전분, 감자 전분 등 포함), 칼슘카보네이트(calcium carbonate), 수크로스(sucrose), 락토오스(lactose), 덱스트로오스, 솔비톨, 만니톨, 자일리톨, 에리스리톨 말티톨, 셀룰로즈, 메틸 셀룰로즈, 나트륨 카르복시메틸셀룰로오즈 및 하이드록시프로필메틸-셀룰로즈 또는 젤라틴 등을 섞어 조제될 수 있다. 예컨대, 활성성분을 고체 부형제와 배합한 다음 이를 분쇄하고 적합한 보조제를 첨가한 후 과립 혼합물로 가공함으로써 정제 또는 당의정제를 수득할 수 있다. 단순한 부형제 이외에 마그네슘 스티레이트 탈크 같은 윤활제들도 사용된다.In addition, the food composition of the present invention may be provided as a solid preparation for oral administration, and examples include tablets, pills, powders, granules, liquids, gels, syrups, slurries, suspensions or capsules, and such solid preparations At least one or more excipients, for example, starch (including corn starch, wheat starch, rice starch, potato starch, etc.), calcium carbonate, sucrose, lactose, dextrose, sorbitol, It can be prepared by mixing mannitol, xylitol, erythritol maltitol, cellulose, methyl cellulose, sodium carboxymethyl cellulose and hydroxypropylmethyl-cellulose or gelatin. For example, tablets or dragees can be obtained by blending the active ingredient with a solid excipient, grinding it, adding suitable adjuvants, and processing it into a granule mixture. In addition to simple excipients, lubricants such as magnesium stearate talc are also used.
또한 본 발명의 식품 조성물은 경구를 위한 액상 제제로 제공될 수 있는데, 일 예로 현탁제, 내용액제, 유제 또는 시럽제 등이 있으며, 흔히 사용되는 단순 희석제인 물 또는 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제 또는 보존제 등이 포함될 수 있다. 또한, 경우에 따라 가교결합 폴리비닐피롤리돈, 한천, 알긴산 또는 나트륨 알기네이트 등을 붕해제로 첨가할 수 있으며, 항응집제, 윤활제, 습윤제, 향료, 유화제 및 방부제 등을 추가로 포함할 수 있다,In addition, the food composition of the present invention may be provided as a liquid formulation for oral use, for example, there are suspensions, solutions, emulsions or syrups, etc., and various excipients, such as water or liquid paraffin, which are commonly used simple diluents, For example, a wetting agent, a sweetening agent, a flavoring agent, or a preservative may be included. In addition, in some cases, cross-linked polyvinylpyrrolidone, agar, alginic acid or sodium alginate may be added as a disintegrant, and an anti-aggregating agent, lubricant, wetting agent, flavoring agent, emulsifying agent and preservative may be additionally included. ,
본 발명의 식품 조성물은 피부 주름 개선 또는 완화, 피부 미백, 피부 보습 효과를 가질 수 있는 것에 사용될 수 있으며, 기능성 식품(functional food), 영양 보조제(nutritional supplement), 건강식품(health food), 식품 첨가제(food additives) 등의 모든 형태를 포함한다. 상기 유형의 식품 조성물은 당 업계에 공지된 통상적인 방법에 따라 다양한 형태로 제조할 수 있다. 일반적인 식품으로는 육류, 곡류, 카페인 음료, 일반음료, 초콜렛, 빵류, 스넥류, 과자류, 피자, 젤리, 면류, 껌류, 아이스크림류, 알코올성 음료, 술, 비타민 복합제 및 그 밖의 건강보조식품류 중 선택되는 어느 하나일 수 있으며, 반드시 이에 한정되는 것은 아니다. 영양보조제로는 이에 한정되지 않지만 캡슐, 타블렛, 환 등에 본 발명의 천연소재추출물 또는 천연소재복합추출물을 첨가하여 제조할 수 있다. 또한, 건강기능식품으로는 이에 한정되지 않지만 예를 들면, 본 발명의 천연소재추출물 또는 천연소재복합추출물 자체를 차, 쥬스 및 드링크의 형태로 제조하여 음용(건강음료)할 수 있도록 액상화, 과립화, 캡슐화 및 분말화하여 섭취할 수 있다. 또한, 본 발명의 천연소재추출물 또는 천연소재복합추출물을 식품 첨가제의 형태로 사용하기 위해서는 분말 또는 농축액 형태로 제조하여 사용할 수 있다.The food composition of the present invention may be used for improving or alleviating skin wrinkles, skin whitening, and skin moisturizing effects, and may be used as a functional food, a nutritional supplement, a health food, or a food additive. (food additives) and the like. Food compositions of this type can be prepared in various forms according to conventional methods known in the art. Common foods include meat, grains, caffeinated beverages, general drinks, chocolate, breads, snacks, confectionery, pizza, jelly, noodles, gum, ice cream, alcoholic beverages, alcohol, vitamin complexes, and other health supplements. It may be one, but is not necessarily limited thereto. The nutritional supplement is not limited thereto, but it can be prepared by adding the natural material extract or natural material complex extract of the present invention to capsules, tablets, pills, and the like. In addition, the health functional food is not limited thereto, but for example, the natural material extract or the natural material complex extract of the present invention is prepared in the form of tea, juice, and drink, and liquefied and granulated so that it can be consumed (health drink). , can be ingested by encapsulation and powder In addition, in order to use the natural material extract or the natural material complex extract of the present invention in the form of a food additive, it may be prepared and used in the form of a powder or a concentrate.
또한, 본 발명의 천연소재추출물 또는 천연소재복합추출물과 항산화, 미백 및 주름 개선, 피부 보습 효과가 있다고 알려진 공지의 활성 성분과 함께 혼합하여 조성물의 형태로 제조할 수 있다.In addition, it can be prepared in the form of a composition by mixing the natural material extract or natural material complex extract of the present invention with known active ingredients known to have antioxidant, whitening and wrinkle improvement, and skin moisturizing effects.
본 발명의 천연소재추출물 또는 천연소재복합추출물은, 식품 조성물 전체에 대하여 0.001~30 중량% 포함되는 것이 좋다. 이보다 적게 첨가될 경우 피부 개선 효과가 미미할 수 있으며, 이를 초과하여 첨가될 경우 첨가량 대비 효과가 경제적으로 바람직하지 못할 수 있으며, 제형 안정성에 문제가 발생할 수 있다.The natural material extract or natural material complex extract of the present invention is preferably contained in an amount of 0.001 to 30% by weight based on the whole food composition. When it is added less than this, the skin improvement effect may be insignificant, and if it is added in excess, the effect may be economically undesirable compared to the added amount, and a problem may occur in formulation stability.
이하, 본 발명에 대해 하기 실시예 및 실험예에서 더욱 상세히 설명하고자 한다. 다만, 본 발명의 권리범위가 하기 실시예 및 실험예에만 한정되는 것은 아니고, 이와 등가의 기술적 사상의 변형까지를 모두 포함한다.Hereinafter, the present invention will be described in more detail in the following Examples and Experimental Examples. However, the scope of the present invention is not limited only to the following examples and experimental examples, and includes all modifications of the technical idea equivalent thereto.
[실시예 1 : 도라지에 포함된 폴리페놀 추출을 위한 최적 추출조건 확립][Example 1: Establishment of optimal extraction conditions for extracting polyphenols contained in bellflower]
껍질을 벗겨 깨끗이 세척 후 건조된 도라지 100g에 포함된 폴리페놀을 최대 효율로 추출하기 위해 반응표면분석법을 통해 추출 조건을 확인하였다. 추출용매는 정제수를 이용하였으며, 건조된 도라지 중량의 10배의 정제수를 가하여 추출하였다. 반응표면분석법의 결과를 표 1 내지 4에 나타내었다.The extraction conditions were checked through reaction surface analysis to extract polyphenols contained in 100 g of dried bellflower after peeling and cleaning with maximum efficiency. Purified water was used as the extraction solvent, and purified
폴리페놀
(mg/g)Bellflower
polyphenols
(mg/g)
도 1에 반응표면분석법에 따른 등고선도와 표면도를 나타내었다. 도 2에서 보듯이, 도라지의 폴리페놀을 최대 효율로 추출할 수 있는 추출조건은 53.7868 ℃, 27.7279 h이며, 폴리페놀은 29.69 mg/g 추출되는 점을 확인하였다.1 shows a contour diagram and a surface diagram according to the response surface analysis method. As shown in FIG. 2, it was confirmed that the extraction conditions for extracting the polyphenols of bellflower with maximum efficiency were 53.7868 °C and 27.7279 h, and 29.69 mg/g of polyphenols were extracted.
[실시예 2 : 배에 포함된 폴리페놀 추출을 위한 최적 추출조건 확립][Example 2: Establishment of optimal extraction conditions for extracting polyphenols contained in pears]
과피를 제거하지 않고 깨끗이 세척 후 건조된 배 100g에 포함된 폴리페놀을 최대 효율로 추출하기 위해 반응표면분석법을 통해 추출 조건을 확인하였다. 추출용매는 정제수를 이용하였으며, 건조된 배 중량의 10배의 정제수를 가하여 추출하였다. 반응표면분석법의 결과를 표 5 내지 8에 나타내었다.The extraction conditions were checked through reaction surface analysis to extract polyphenols contained in 100 g of dried pears with maximum efficiency after washing without removing the skin. Purified water was used as the extraction solvent, and purified
(mg/g)pear polyphenols
(mg/g)
도 3에 반응표면분석법에 따른 등고선도와 표면도를 나타내었다. 도 4에서 보듯이, 배의 폴리페놀을 최대 효율로 추출할 수 있는 추출조건은 96.2132 ℃, 20.0140 h이며, 폴리페놀은 35.84 mg/g 추출되는 점을 확인하였다.3 shows a contour diagram and a surface diagram according to the response surface analysis method. As shown in FIG. 4, it was confirmed that the extraction conditions for extracting polyphenols of pears with maximum efficiency were 96.2132 °C and 20.0140 h, and 35.84 mg/g of polyphenols were extracted.
[실시예 3 : 석류에 포함된 폴리페놀 추출을 위한 최적 추출조건 확립][Example 3: Establishment of optimal extraction conditions for extraction of polyphenols contained in pomegranate]
깨끗이 세척 후 건조된 석류의 열매(겉껍질 제거, 씨를 포함) 100g에 포함된 폴리페놀을 최대 효율로 추출하기 위해 반응표면분석법을 통해 추출 조건을 확인하였다. 추출용매는 정제수를 이용하였으며, 건조된 석류 중량의 10배의 정제수를 가하여 추출하였다. 반응표면분석법의 결과를 표 9 내지 12에 나타내었다.To extract polyphenols contained in 100 g of dried pomegranate fruit (including peel and seeds) with maximum efficiency after washing, the extraction conditions were confirmed through reaction surface analysis. Purified water was used as the extraction solvent, and purified
폴리페놀
(mg/g)Pomegranate
polyphenols
(mg/g)
도 5에 반응표면분석법에 따른 등고선도와 표면도를 나타내었다. 도 6에서 보듯이, 석류의 폴리페놀을 최대 효율로 추출할 수 있는 추출조건은 96.2132 ℃, 14.1000 h이며, 폴리페놀은 37.31 mg/g 추출되는 점을 확인하였다.5 shows a contour diagram and a surface diagram according to the response surface analysis method. As shown in FIG. 6 , it was confirmed that the extraction conditions for extracting polyphenols of pomegranate with maximum efficiency were 96.2132° C. and 14.1000 h, and 37.31 mg/g of polyphenols were extracted.
[실시예 4 : 자소엽에 포함된 폴리페놀 추출을 위한 최적 추출조건 확립][Example 4: Establishment of optimal extraction conditions for extraction of polyphenols contained in perilla leaves]
깨끗이 세척 후 건조된 자소엽 100g에 포함된 폴리페놀을 최대 효율로 추출하기 위해 반응표면분석법을 통해 추출 조건을 확인하였다. 추출용매는 정제수를 이용하였으며, 건조된 자소엽 중량의 10배의 정제수를 가하여 추출하였다. 반응표면분석법의 결과를 표 13 내지 16에 나타내었다.In order to extract polyphenols contained in 100 g of dried perilla leaves after cleaning with maximum efficiency, the extraction conditions were confirmed through reaction surface analysis. Purified water was used as the extraction solvent, and purified
폴리페놀
(mg/g)perilla leaf
polyphenols
(mg/g)
도 7에 반응표면분석법에 따른 등고선도와 표면도를 나타내었다. 도 8에서 보듯이, 자소엽의 폴리페놀을 최대 효율로 추출할 수 있는 추출조건은 53.7868 ℃, 27.7279 h이며, 폴리페놀은 33.02 mg/g 추출되는 점을 확인하였다.7 shows a contour diagram and a surface diagram according to the response surface analysis method. As shown in FIG. 8, it was confirmed that the extraction conditions for extracting polyphenols from perilla leaves with maximum efficiency were 53.7868 °C and 27.7279 h, and 33.02 mg/g of polyphenols were extracted.
[실험예 1 : 도라지, 배, 석류, 자소엽 단독 처리 시 세포생존률 측정] [Experimental Example 1: Measurement of cell viability when treated with bellflower, pear, pomegranate and perilla leaf alone]
본 실험에서는 상기 실시예 1 내지 4에서 확립한 각 소재의 추출조건에 따라 제조된 추출물의 세포독성을 평가하였다. 각 추출물은 추출 후 감압농축 및 동결건조하여 고형화하였다.In this experiment, the cytotoxicity of the extract prepared according to the extraction conditions of each material established in Examples 1 to 4 was evaluated. After extraction, each extract was concentrated under reduced pressure and lyophilized to solidify.
세포실험시 시료가 세포에 미치는 독성으로 인해 핵, 미토콘드리아, 시토졸 등에 영향을 미쳐 부정확한 결과가 도출될 수 있으므로 세포생존률에 영향을 미치지 않는 조건(세포생존률 80% 이상)의 농도에서 실험을 진행하였다. 세포생존률 시험은 B16-F10(마우스 유래 흑색종 세포), WI-38(인간유래 섬유아세포), HaCaT(각질 형성 세포) 세포에 대하여 진행하였다. 각 세포를 배양하고 96 well에 1×104 cell로 분주한 뒤 24시간 배양하였다. 이후 도라지추출물, 배추출물, 석류추출물, 자소엽추출물을 각각 0.1, 0.25, 0.5 및 1 mg/ml 처리하고 이를 24시간 배양하였다. 이후 배지를 제거하고 PBS로 세척한 뒤 MTT reagent (0.5mg/ml)를 처리한 후 4시간 배양하한 뒤, 이를 PBS로 세척 후 건조하였다. 건조 후 DMSO를 100㎕ 처리하여 formazan을 용출시키고 570nm에서 흡광도를 측정하였다.During cell experiments, the toxicity of the sample on cells may affect the nucleus, mitochondria, and cytosol, resulting in inaccurate results. did. Cell viability tests were performed on B16-F10 (mouse-derived melanoma cells), WI-38 (human-derived fibroblasts), and HaCaT (keratinocytes) cells. Each cell was cultured, and 1×10 4 cells were aliquoted into 96 wells and cultured for 24 hours. Afterwards, 0.1, 0.25, 0.5 and 1 mg/ml of bellflower extract, pear extract, pomegranate extract, and perilla leaf extract were treated, respectively, and cultured for 24 hours. Thereafter, the medium was removed, washed with PBS, treated with MTT reagent (0.5 mg/ml), incubated for 4 hours, washed with PBS, and dried. After drying, 100 μl of DMSO was treated to elute formazan, and absorbance was measured at 570 nm.
도 9는 B16-F10 세포의 세포생존률을 확인한 결과이다. 본 발명의 도라지추출물, 배추출물, 석류추출물, 자소엽추출물을 농도별로 처리한 결과 B16-F10 세포생존률에 영향을 미치지 않는 점을 확인하였으며, 이에 따라 본 발명의 천연소재추출물은 0.1, 0.25, 0.5 및 1 mg/ml 농도 범위에서 세포독성을 나타내지 않는 점을 확인하였다.9 is a result of confirming the cell viability of B16-F10 cells. As a result of treating the bellflower extract, pear extract, pomegranate extract, and perilla leaf extract of the present invention by concentration, it was confirmed that the B16-F10 cell viability was not affected. Accordingly, the natural material extract of the present invention was 0.1, 0.25, 0.5 And it was confirmed that it does not exhibit cytotoxicity in the concentration range of 1 mg/ml.
도 10은 WI-38 세포의 세포생존률을 확인한 결과이다. 본 발명의 도라지추출물, 배추출물, 석류추출물, 자소엽추출물을 농도별로 처리한 결과 WI-38 세포생존률에 영향을 미치지 않는 점을 확인하였으며, 이에 따라 본 발명의 천연소재추출물은 0.1, 0.25, 0.5 및 1 mg/ml 농도 범위에서 세포독성을 나타내지 않는 점을 확인하였다.10 is a result confirming the cell viability of WI-38 cells. As a result of treating the bellflower extract, pear extract, pomegranate extract, and perilla leaf extract of the present invention by concentration, it was confirmed that the WI-38 cell viability was not affected. Accordingly, the natural extract of the present invention was 0.1, 0.25, 0.5 And it was confirmed that it does not exhibit cytotoxicity in the concentration range of 1 mg/ml.
도 11은 HaCaT 세포의 세포생존률을 확인한 결과이다. 본 발명의 도라지추출물, 배추출물, 석류추출물, 자소엽추출물을 농도별로 처리한 결과 HaCaT 세포생존률에 영향을 미치지 않는 점을 확인하였으며, 이에 따라 본 발명의 천연소재추출물은 0.1, 0.25, 0.5 및 1 mg/ml 농도 범위에서 세포독성을 나타내지 않는 점을 확인하였다.11 is a result confirming the cell viability of HaCaT cells. As a result of treating the bellflower extract, pear extract, pomegranate extract, and perilla leaf extract of the present invention by concentration, it was confirmed that the HaCaT cell viability was not affected. Accordingly, the natural material extract of the present invention was 0.1, 0.25, 0.5 and 1 It was confirmed that there was no cytotoxicity in the mg/ml concentration range.
[실험예 2 : 도라지, 배, 석류, 자소엽 단독 처리 시 멜라닌 생성 억제능 확인] [Experimental Example 2: Confirmation of melanin production inhibitory ability when treated with bellflower, pear, pomegranate, and perilla leaves alone]
자외선 등의 외부 환경에 의해 멜라닌이 비정상적으로 증가함에 따라 피부에 기미 또는 색소침착이 발생한다. 본 발명은 상기 실시예 1 내지 4의 최적 조건에 따라 제조된 도라지추출물, 배추출물, 석류추출물, 자소엽추출물이 각각 멜라닌 생성을 억제하여 피부 미백 효과를 발휘하는지 확인하였다. 대조군으로는 알부틴 0.01 μM을 사용하였다.As melanin increases abnormally by external environment such as ultraviolet rays, blemishes or pigmentation occurs on the skin. In the present invention, it was confirmed whether the bellflower extract, pear extract, pomegranate extract, and perilla leaf extract prepared according to the optimal conditions of Examples 1 to 4 respectively inhibit melanin production to exert a skin whitening effect. As a control, 0.01 μM of arbutin was used.
마우스 유래 흑색종세포인 B16-F10 세포를 배양하고 본 발명의 도라지추출물, 배추출물, 석류추출물, 자소엽추출물을 각각 1 mg/ml 처리한 뒤, 100 nM의 α-MSH를 처리하고 24시간 배양 후 배지를 제거한 뒤 PBS로 세척하였다. 1N NaOH 100 ㎕를 처리한 후 60 ℃에서 1시간 멜라닌을 추출하고 405 nm에서 흡광도를 측정하였다(For each group, n=3±SD, **p<0.005, ***p<0.001 compared by control).B16-F10 cells, which are mouse-derived melanoma cells, were cultured, treated with 1 mg/ml each of the bellflower extract, pear extract, pomegranate extract, and perilla leaf extract of the present invention, treated with 100 nM α-MSH and cultured for 24 hours. After removing the medium, it was washed with PBS. After treatment with
도 12에서 보듯이, 본 발명의 도라지추출물과 배추출물이 α-MSH에 유도된 멜라닌 생성을 억제하는 점을 확인하였으며, 특히 대조군과 유사한 멜라닌 생성 억제 효과를 보이는 점을 확인하였다.As shown in FIG. 12 , it was confirmed that the bellflower extract and the Chinese cabbage extract of the present invention inhibited α-MSH-induced melanin production, and in particular, it was confirmed that it showed a similar melanin production inhibitory effect to that of the control group.
[실험예 3 : 도라지, 배, 석류, 자소엽 단독 처리 시 프로콜라겐 생성능 확인] [Experimental Example 3: Confirmation of procollagen production ability when treated with bellflower, pear, pomegranate, and perilla leaves alone]
피부의 진피층은 섬유아세포, 콜라겐, 엘라스틴이 유기적으로 구성되어 있으나, 노화, 자외선 등의 피부 손상에 의해 콜라겐, 엘라스틴 등의 유기구조가 붕괴되면서 주름이 생성된다. 프로콜라겐(procollagen)은 콜라겐으로 전환되어 진피층의 콜라겐 함량을 증가시켜 주름 개선 또는 완화 효과를 발휘한다. 본 발명은 상기 실시예 1 내지 4의 최적 조건에 따라 제조된 도라지추출물, 배추출물, 석류추출물, 자소엽추출물이 각각 프로콜라겐 생성을 촉진하여 피부 주름 개선 효과를 발휘하는지 확인하였다. 대조군으로는 레티놀 0.5 μM을 사용하였다.The dermal layer of the skin is organically composed of fibroblasts, collagen, and elastin, but wrinkles are generated as the organic structures such as collagen and elastin are destroyed by skin damage such as aging and ultraviolet rays. Procollagen is converted to collagen and increases the collagen content in the dermal layer, thereby improving or alleviating wrinkles. In the present invention, it was confirmed whether the bellflower extract, pear extract, pomegranate extract, and perilla leaf extract prepared according to the optimal conditions of Examples 1 to 4, respectively, promote procollagen production and thus exert the effect of improving skin wrinkles. As a control, 0.5 μM of retinol was used.
인간유래 섬유아세포인 WI-38 세포를 배양하고 본 발명의 도라지추출물, 배추출물, 석류추출물, 자소엽추출물을 각각 1 mg/ml 처리한 뒤 24시간 배양 후 배지를 회수하였다. Antibody-POD Conjugatd Solution 100 ㎕를 항체가 코팅된 96 well에 분주하고, 배지와 standard solution 20 ㎕를 각 well에 분주한 뒤 37℃에서 3시간 반응시켰다. Wash buffer 400 ㎕로 4회 세척 후, Substrate Solution (TMBZ) 100 ㎕를 각 well에 분주하고 실온에서 15분 반응시켰다. Stop solution 100 ㎕를 각 well에 분주한 뒤 450 nm에서 흡광도를 측정하였다(For each group, n=3±SD, **p<0.005, ***p<0.001 compared by control).Human-derived fibroblasts, WI-38 cells, were cultured, treated with 1 mg/ml each of the bellflower extract, pear extract, pomegranate extract, and perilla leaf extract of the present invention, and cultured for 24 hours, then the medium was recovered. 100 μl of Antibody-POD Conjugatd Solution was dispensed into 96 wells coated with antibody, and 20 μl of medium and standard solution were dispensed into each well and reacted at 37° C. for 3 hours. After washing 4 times with 400 μl of wash buffer, 100 μl of Substrate Solution (TMBZ) was dispensed into each well and reacted at room temperature for 15 minutes. 100 μl of Stop solution was dispensed into each well and absorbance was measured at 450 nm (For each group, n=3±SD, **p<0.005, ***p<0.001 compared by control).
도 13에서 보듯이, 본 발명의 도라지추출물, 배추출물, 석류추출물이 프로콜라겐 생성 촉진 효과가 우수한 점을 확인하였으며, 특히 도라지추출물과 석류추출물은 대조군과 유사한 프로콜라겐 생성능을 보이는 점을 확인하였다.As shown in FIG. 13 , it was confirmed that the bellflower extract, pear extract, and pomegranate extract of the present invention were excellent in promoting procollagen production, and in particular, it was confirmed that the bellflower extract and pomegranate extract showed similar procollagen production ability to that of the control group.
[실험예 4 : 도라지, 배, 석류, 자소엽 단독 처리 시 히알루론산 생성능 확인] [Experimental Example 4: Confirmation of hyaluronic acid production ability when treated with bellflower, pear, pomegranate, and perilla leaf alone]
각질세포는 세포간 지질과 함수율을 통해 피부의 유수분 균형을 유지한다. 세포 내 지질의 양이 감소하거나 수분이 부족하면 각질이 발생하거나 피부염증 등의 문제가 발생하므로 적절한 유수분 조절이 필요하다. 본 발명은 상기 실시예 1 내지 4의 최적 조건에 따라 제조된 도라지추출물, 배추출물, 석류추출물, 자소엽추출물이 각각 히알루론산(hyaluronic acid) 생성을 촉진하여 피부 유수분 균형 유지, 피부 보습 효과를 발휘하는지 확인하였다. 대조군으로는 GlcNAc(N-Acetylglucosamine) 0.1 μM을 사용하였다.Keratinocytes maintain the oil-water balance of the skin through intercellular lipids and moisture content. When the amount of intracellular lipids decreases or moisture is insufficient, keratin occurs or problems such as skin inflammation occur, so proper oil and moisture control is required. According to the present invention, the bellflower extract, pear extract, pomegranate extract, and perilla leaf extract prepared according to the optimal conditions of Examples 1 to 4 promote the production of hyaluronic acid, respectively, to maintain the skin oil-water balance and to maintain the skin moisturizing effect. confirmed that it does. As a control, 0.1 μM of GlcNAc (N-Acetylglucosamine) was used.
인간유래 각질세포인 HaCaT 세포를 배양하고 본 발명의 도라지추출물, 배추출물, 석류추출물, 자소엽추출물을 각각 1 mg/ml 처리한 뒤 24시간 배양 후 세포를 회수하였다. Sonicator를 이용하여 세포를 용출(lysis)시키고 단백질을 정량하였다. 96 well plate에 세포 추출액 50 ㎕와 Assy Diluent RD1-14 50 ㎕를 분주하고 실온에서 2시간 반응시킨 후 Wash buffer를 이용하여 5회 세척하였다. Hyaluronan conjugate 100 ㎕를 분주하고 실온에서 2시간 반응시킨 후 Wash buffer를 이용하여 5회 세척하였다. Substrate solution 100 ㎕를 분주하고 30분 반응시킨 뒤 Stop solution 100 ㎕를 처리하여 반응을 종결시켰다. 이를 405 nm에서 흡광도를 측정하였다(For each group, n=3±SD, **p<0.005, ***p<0.001 compared by control).HaCaT cells, which are human-derived keratinocytes, were cultured, treated with 1 mg/ml each of the bellflower extract, pear extract, pomegranate extract, and perilla leaf extract of the present invention, and the cells were recovered after culturing for 24 hours. Cells were lysed using a sonicator and protein was quantified. In a 96 well plate, 50 μl of the cell extract and 50 μl of Assy Diluent RD1-14 were dispensed, reacted at room temperature for 2 hours, and washed 5 times using Wash buffer. 100 μl of hyaluronan conjugate was dispensed and reacted at room temperature for 2 hours, followed by washing 5 times using wash buffer. After dispensing 100 μl of the substrate solution and reacting for 30 minutes, 100 μl of Stop solution was treated to terminate the reaction. The absorbance was measured at 405 nm (For each group, n=3±SD, **p<0.005, ***p<0.001 compared by control).
도 14에서 보듯이, 본 발명의 도라지추출물, 배추출물, 석류추출물, 자소엽추출물은 히알루론산 생성 효과를 보이는 점을 확인하였으며, 특히 배추출물과 자소엽추출물이 히알루론산 생성 효과가 우수한 점을 확인하였다.As shown in FIG. 14, it was confirmed that the bellflower extract, pear extract, pomegranate extract, and perilla leaf extract of the present invention showed hyaluronic acid production effect, and in particular, it was confirmed that the cabbage extract and perilla leaf extract had excellent hyaluronic acid production effect. did.
[실시예 5 : 도라지, 배, 석류, 자소엽 복합추출물의 제조] [Example 5: Preparation of bellflower, pear, pomegranate, perilla leaf complex extract]
상기 실시예 1 내지 4의 최적 조건에 따라 제조된 도라지추출물, 배추출물, 석류추출물, 자소엽추출물을 아래 표 17과 같은 중량비로 혼합하여 복합추출물을 제조하였다.The bellflower extract, pear extract, pomegranate extract, and perilla leaf extract prepared according to the optimal conditions of Examples 1 to 4 were mixed in the weight ratio as shown in Table 17 below to prepare a complex extract.
[실험예 5 : 복합추출물의 멜라닌 생성 억제능 확인] [Experimental Example 5: Confirmation of melanin production inhibitory ability of complex extract]
본 실험은 상기 실시예 5에서 제조한 복합추출물의 멜라닌 생성 억제능을 확인하기 위해 복합추출물을 1 mg/ml 처리하여 상기 실험예 2와 동일하게 실험하였다. 대조군으로는 알부틴 0.01 μM을 사용하였다.In this experiment, 1 mg/ml of the complex extract was treated in the same manner as in Experimental Example 2 to confirm the melanin production inhibitory ability of the complex extract prepared in Example 5. As a control, 0.01 μM of arbutin was used.
도 15에서 보듯이, 실시예 5-1 내지 5-11 모두에서 멜라닌 생성 억제 효과를 보였으며, 특히 실시예 5-1, 5-8, 5-11은 우수한 멜라닌 생성 억제 효과를 보였다.As shown in Figure 15, all of Examples 5-1 to 5-11 showed a melanin production inhibitory effect, in particular Examples 5-1, 5-8, 5-11 showed an excellent melanogenesis inhibitory effect.
상기 실험예 2에서 석류추출물과 자소엽추출물이 멜라닌 생성 억제 효과를 보이지 않았음에도 불구하고, 도라지추출물, 배추출물, 석류추출물, 자소엽추출물을 복합물로 처리함에 따라 멜라닌 생성 억제 효과, 즉 피부 미백효과를 보이는 점을 확인하였다.Although the pomegranate extract and perilla leaf extract showed no melanin production inhibitory effect in Experimental Example 2, the melanin production inhibitory effect, that is, the skin whitening effect, was treated with the bellflower extract, pear extract, pomegranate extract, and perilla leaf extract with the complex. was confirmed to be visible.
[실험예 6 : 복합추출물의 프로콜라겐 생성능 확인] [Experimental Example 6: Confirmation of procollagen production ability of complex extract]
본 실험은 상기 실시예 5에서 제조한 복합추출물의 프로콜라겐 생성능을 확인하기 위해 복합추출물을 1 mg/ml 처리하여 상기 실험예 3과 동일하게 실험하였다. 대조군으로는 레티놀 0.5 μM을 사용하였다.In this experiment, 1 mg/ml of the complex extract was treated in the same manner as in Experimental Example 3 to confirm the procollagen production ability of the complex extract prepared in Example 5. As a control, 0.5 μM of retinol was used.
도 16에서 보듯이, 실시예 5-2와 실시예 5-5를 제외한 모든 군에서 프로콜라겐 생성 효과를 확인하였으며, 특히 실시예 5-1, 5-3 및 5-10은 우수한 프로콜라겐 생성 효과를 보였다.As shown in FIG. 16 , the procollagen generating effect was confirmed in all groups except Examples 5-2 and 5-5, and in particular, Examples 5-1, 5-3 and 5-10 had excellent procollagen generating effect. showed
상기 실험예 3에서 자소엽추출물이 프로콜라겐 생성 효과를 보이지 않았고, 비추출물이 상대적으로 낮은 프로콜라겐 생성 효과를 보였음에도 불구하고, 도라지추출물, 배추출물, 석류추출물, 자소엽추출물을 복합물로 처리함에 따라 프로콜라겐 생성 효과, 즉 피부 주름 개선 또는 완화 효과를 보이는 점을 확인하였다.In Experimental Example 3, although the perilla leaf extract showed no procollagen-generating effect and the non-extract showed a relatively low procollagen-generating effect, bellflower extract, pear extract, pomegranate extract, and perilla leaf extract were treated with a complex Accordingly, it was confirmed that the procollagen production effect, that is, the skin wrinkle improvement or alleviation effect was shown.
[실험예 7 : 복합추출물의 히알루론산 생성능 확인] [Experimental Example 7: Confirmation of hyaluronic acid production ability of complex extract]
본 실험은 상기 실시예 5에서 제조한 복합추출물의 히알루론산 생성능을 확인하기 위해 복합추출물을 1 mg/ml 처리하여 상기 실험예 4와 동일하게 실험하였다. 대조군으로는 GlcNAc(N-Acetylglucosamine) 0.1 μM을 사용하였다.In this experiment, 1 mg/ml of the complex extract was treated in the same manner as in Experimental Example 4 to confirm the hyaluronic acid production ability of the complex extract prepared in Example 5. As a control, 0.1 μM of GlcNAc (N-Acetylglucosamine) was used.
도 17에서 보듯이, 실시예 5-3과 실시예 5-5를 제외한 모든 군에서 히알루론산 생성 효과를 확인하였으며, 특히 실시예 5-1, 5-4, 5-6 및 5-9는 우수한 히알루론산 생성 효과를 보였다.As shown in FIG. 17 , the hyaluronic acid production effect was confirmed in all groups except Examples 5-3 and 5-5, and in particular, Examples 5-1, 5-4, 5-6 and 5-9 were excellent It showed the effect of producing hyaluronic acid.
상기 실험예 4에서 도라지추출물과 석류추출물이 상대적으로 낮은 히알루론산 생성 효과를 보였음에도 불구하고, 도라지추출물, 배추출물, 석류추출물, 자소엽추출물을 복합물로 처리함에 따라 히알루론산 생성 효과, 즉 피부 유수분 균형 유지 효과 및 피부 보습 효과를 보이는 점을 확인하였다.Although the bellflower extract and the pomegranate extract showed a relatively low hyaluronic acid production effect in Experimental Example 4, the bellflower extract, the Chinese cabbage extract, the pomegranate extract, and the perilla leaf extract were treated with the complex to produce hyaluronic acid, that is, the skin oil and moisture content. It was confirmed that it showed the effect of maintaining the balance and moisturizing the skin.
Claims (8)
상기 도라지추출물은 51~55 ℃에서 25~29 시간 추출한 것이고,
상기 배추출물은 94~98 ℃에서 18~22 시간 추출한 것이고,
상기 석류추출물은 94~98 ℃에서 12~16 시간 추출한 것이고,
상기 자소엽추출물은 51~55 ℃에서 25~29 시간 추출한 것이며,
도라지추출물, 배추출물, 석류추출물 및 자소엽추출물은 1:1:1:1의 중량비, 1:2:1:1의 중량비 또는 1:2:1:2의 중량비로 혼합되는 것을 특징으로 하는 폴리페놀이 강화된 피부 주름 개선용 식품 조성물.
Contains all of bellflower extract, pear extract, pomegranate extract and perilla leaf extract,
The bellflower extract was extracted at 51-55 ℃ for 25-29 hours,
The pear extract was extracted at 94-98 ℃ for 18-22 hours,
The pomegranate extract is extracted at 94-98 ℃ for 12-16 hours,
The perilla leaf extract was extracted at 51-55 ℃ for 25-29 hours,
Poly, characterized in that bellflower extract, pear extract, pomegranate extract and perilla leaf extract are mixed in a weight ratio of 1:1:1:1, 1:2:1:1 or 1:2:1:2 by weight. A food composition for improving skin wrinkles fortified with phenol.
상기 도라지추출물은 51~55 ℃에서 25~29 시간 추출한 것이고,
상기 배추출물은 94~98 ℃에서 18~22 시간 추출한 것이고,
상기 석류추출물은 94~98 ℃에서 12~16 시간 추출한 것이고,
상기 자소엽추출물은 51~55 ℃에서 25~29 시간 추출한 것이며,
도라지추출물, 배추출물, 석류추출물 및 자소엽추출물은 1:1:1:1의 중량비, 2:1:1:2의 중량비 또는 1:1:2:2의 중량비로 혼합되는 것을 특징으로 하는 폴리페놀이 강화된 피부 미백용 식품 조성물.
Contains all of bellflower extract, pear extract, pomegranate extract and perilla leaf extract,
The bellflower extract was extracted at 51-55 ℃ for 25-29 hours,
The pear extract was extracted at 94-98 ℃ for 18-22 hours,
The pomegranate extract is extracted at 94-98 ℃ for 12-16 hours,
The perilla leaf extract was extracted at 51-55 ℃ for 25-29 hours,
Poly, characterized in that bellflower extract, pear extract, pomegranate extract and perilla leaf extract are mixed in a weight ratio of 1:1:1:1, 2:1:1:2 or 1:1:2:2 by weight. A food composition for skin whitening fortified with phenol.
상기 도라지추출물은 51~55 ℃에서 25~29 시간 추출한 것이고,
상기 배추출물은 94~98 ℃에서 18~22 시간 추출한 것이고,
상기 석류추출물은 94~98 ℃에서 12~16 시간 추출한 것이고,
상기 자소엽추출물은 51~55 ℃에서 25~29 시간 추출한 것이며,
도라지추출물, 배추출물, 석류추출물 및 자소엽추출물은 1:1:1:1의 중량비, 1:1:2:1의 중량비, 2:2:1:1의 중량비 또는 1:2:2:1의 중량비로 혼합되는 것을 특징으로 하는 폴리페놀이 강화된 피부 보습용 식품 조성물.
Contains all of bellflower extract, pear extract, pomegranate extract and perilla leaf extract,
The bellflower extract was extracted at 51-55 ℃ for 25-29 hours,
The pear extract was extracted at 94-98 ℃ for 18-22 hours,
The pomegranate extract is extracted at 94-98 ℃ for 12-16 hours,
The perilla leaf extract was extracted at 51-55 ℃ for 25-29 hours,
Bellflower extract, pear extract, pomegranate extract, and perilla leaf extract have a weight ratio of 1:1:1:1, a weight ratio of 1:1:2:1, a weight ratio of 2:2:1:1, or 1:2:2:1 Polyphenol-enhanced skin moisturizing food composition, characterized in that it is mixed in a weight ratio of
상기 도라지추출물, 배추출물, 석류추출물 및 자소엽추출물은,
추출용매로 정제수를 이용한 것을 특징으로 하는 식품 조성물.
According to any one of claims 1 to 3 selected from,
The bellflower extract, pear extract, pomegranate extract and perilla leaf extract,
A food composition, characterized in that purified water is used as an extraction solvent.
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