KR102286969B1 - Composition for improving skin conditions comprising omega-7 derived from refined fish oil and red ginseng - Google Patents
Composition for improving skin conditions comprising omega-7 derived from refined fish oil and red ginseng Download PDFInfo
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- KR102286969B1 KR102286969B1 KR1020190074327A KR20190074327A KR102286969B1 KR 102286969 B1 KR102286969 B1 KR 102286969B1 KR 1020190074327 A KR1020190074327 A KR 1020190074327A KR 20190074327 A KR20190074327 A KR 20190074327A KR 102286969 B1 KR102286969 B1 KR 102286969B1
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- omega
- red ginseng
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- skin
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Abstract
본 발명은 정제어유 유래 오메가-7 및 홍삼을 포함하는 피부 상태 개선용 조성물에 관한 것으로, 본 발명에 따른 조성물은 오메가-7 및 홍삼 복합물을 유효성분으로 함에 따라 우수한 피부 개선 효과를 나타냄을 특징으로 한다. 본 발명에 따른 피부 상태 개선용 조성물은 정제어유 유래 오메가-7 및 사포닌, 다당체 등을 유효성분으로 함유하는 홍삼을 최적화된 농도로 포함하여, 높은 안정성을 기반으로 피부 보습도 개선, 피부보호 장벽 개선 및 산화적 스트레스에 의한 세포손상 저해 효과를 가져 미세먼지와 같은 외부 자극에 의한 피부의 손상에 대하여 종합적인 피부 상태 개선 효과를 발휘한다. 뿐만 아니라, 항산화, 항염증, 피부 탄력 및 주름 개선 효과를 가지고 있어 우수한 피부 노화 억제 효과를 나타낸다. 본 발명에 따른 조성물은 종합적으로 근본적인 피부 상태 개선 효과를 나타냄에 따라 식품, 건강기능식품, 식품 첨가물 및 화장품 등으로 다양하게 활용될 것으로 기대된다.The present invention relates to a composition for improving skin condition comprising omega-7 derived from refined fish oil and red ginseng, wherein the composition according to the present invention exhibits excellent skin improvement effect by using omega-7 and red ginseng complex as active ingredients. do. The composition for improving skin condition according to the present invention contains red ginseng containing refined fish oil-derived omega-7 and saponins and polysaccharides as active ingredients in an optimized concentration, improving skin moisture and improving skin protection barrier based on high stability And it has an inhibitory effect on cell damage caused by oxidative stress, thereby exhibiting a comprehensive skin condition improvement effect against damage to the skin caused by external stimuli such as fine dust. In addition, it has antioxidant, anti-inflammatory, skin elasticity and anti-wrinkle effects, thereby showing excellent skin aging inhibitory effects. The composition according to the present invention is expected to be variously used as food, health functional food, food additive, cosmetics, etc. as it comprehensively exhibits a fundamental skin condition improvement effect.
Description
본 발명은 정제어유 유래 오메가-7 및 홍삼을 포함하는 피부 상태 개선용 조성물에 관한 것으로, 본 발명에 따른 조성물은 오메가-7 및 홍삼 복합물을 유효성분으로 함에 따라 우수한 피부 개선 효과를 나타냄을 특징으로 한다.The present invention relates to a composition for improving skin condition comprising omega-7 derived from refined fish oil and red ginseng, wherein the composition according to the present invention exhibits excellent skin improvement effect by using omega-7 and red ginseng complex as active ingredients. do.
피부는 인체의 대부분을 차지하며 체온조절, 분비, 배설, 흡수 등의 다양한 작용을 하고, 특히 인체의 최외부에 존재함으로써 외부로부터 물리적, 화학적, 생물학적으로 인체를 보호하는 기능을 가진다. 피부는 표피, 진피, 피하지방으로 이루어져 있으며, 표피는 인체의 외부와 내부로부터 방어작용을 하고, 진피는 피부의 탄력 및 지지 역할, 피하지방은 체온을 유지하는 기능을 가진다.The skin occupies most of the human body and performs various functions such as thermoregulation, secretion, excretion, and absorption. The skin consists of the epidermis, dermis, and subcutaneous fat. The epidermis acts as a defense against the outside and inside of the human body, the dermis plays a role in elasticity and support of the skin, and the subcutaneous fat maintains body temperature.
피부는 또한 다양한 물리적 및 화학적 외부 자극으로부터 우리 몸을 보호하는 첫 번째 장벽으로 중요한 역할을 하고 있다. 피부는 표피, 진피 및 피하 조직으로 이루어져 있으며, 표피는 주로 각화 세포로 구성되어있다. 이 각화 세포는 자외선, 환경 독소, 열과 같은 외부 자극에 민감한데(Fisher, Kang et al. 2002), 특히 자외선에 의한 자극은 H2O2를 발생시켜 세포 내 산화스트레스의 증가를 야기한다(Halliwell and Gutteridge 1985, Ozben 2007). 그 결과, 피부 재생 단백질과 피부 탄력 단백질의 발현이 감소하여 주름 형성을 촉진시키고 피부 노화를 촉진시킨다(Talwar, Griffiths et al. 1995).The skin also plays an important role as the first barrier that protects our body from a variety of physical and chemical external stimuli. The skin consists of epidermis, dermis, and subcutaneous tissue, and the epidermis is mainly composed of keratinocytes. These keratinocytes are sensitive to external stimuli such as UV rays, environmental toxins, and heat (Fisher, Kang et al. 2002). In particular, stimulation by UV rays generates H 2 O 2 , causing an increase in intracellular oxidative stress (Halliwell). and Gutteridge 1985, Ozben 2007). As a result, the expression of skin regenerating protein and skin elasticity protein is reduced, thereby promoting wrinkle formation and skin aging (Talwar, Griffiths et al. 1995).
콜라겐 분해효소인 기질 금속 단백질 분해 효소(Matrix Metalloproteinase; MMP)라는 효소는 노화방지 및 상처개선에 관여한다. 즉, 생체 내에서 콜라겐과 같은 세포외기질의 합성과 분해는 적절하게 조절되나, 노화가 진행되거나 자외선 조사에 의해 콜라겐 합성이 감소하고, 콜라겐을 분해하는 효소인 기질 금속 단백질 분해 효소(MMP)의 발현이 촉진되어 피부의 탄력이 저하되고 주름이 형성된다. 또한 콜라겐을 분해하는 기질 금속 단백질 분해 효소의 발현이 감소되면 상처가 잘 아물지 않게 된다.An enzyme called Matrix Metalloproteinase (MMP), a collagen degrading enzyme, is involved in anti-aging and wound improvement. That is, the synthesis and decomposition of extracellular matrix such as collagen in vivo is appropriately regulated, but collagen synthesis decreases with aging or UV irradiation, and expression of matrix metalloproteinase (MMP), an enzyme that decomposes collagen This is accelerated, the elasticity of the skin is lowered and wrinkles are formed. In addition, when the expression of matrix metalloproteinase that degrades collagen is reduced, the wound does not heal well.
한편, 활성 산소종은 산소분자가 호흡과정을 통해 불완전하게 환원되어 생기는 매우 활성이 높은 중간산물로, 여러 신호전달 과정을 변경하여 유전자 발현, 세포흡착, 물질대사, 세포주기, 세포사멸에 영향을 주는 것으로 알려져 있으며, 활성 산소종이 유도하는 신호전달에는 MAPK(mitogen-activated protein kinase), NF-κB(nuclear factor-κB), PI3K(phosphatidylinositol 3-kinase), p53, β-catenin/Wnt를 매개로 하는 신호전달이 알려져 있다. 최근, 활성 산소종에 의한 인체 내 손상을 보호하는 항산화를 통한 노화방지에 대한 연구가 다양하게 이루어지고 있다.On the other hand, reactive oxygen species are highly active intermediate products that are produced by incomplete reduction of oxygen molecules through the respiration process. The signaling induced by reactive oxygen species is mediated by mitogen-activated protein kinase (MAPK), nuclear factor-κB (NF-κB), phosphatidylinositol 3-kinase (PI3K), p53, and β-catenin/Wnt. signaling is known. Recently, various studies have been made on anti-aging through antioxidants that protect the damage in the human body caused by reactive oxygen species.
껍질을 벗기지 않은 인삼을 증기로 찐 뒤 건조하여 숙성하는 과정을 반복해 가공한 것을 홍삼(Korean red ginseng)이라 말한다. 홍삼은 중추신경에 대해 진정 및 흥분작용을 나타내며, 순환계에 작용하여 고혈압 또는 동맥경화에 대한 예방효과를 나타낸다. 이에 더해 조혈작용과 혈당을 저하시키며, 간의 보호, 내분비계에 작용해 성행동이나 생식효과에 간접적인 유효 작용을 하며, 항염 또는 항종양작용까지 있고, 피부를 보호하고 부드럽게 하는 효과도 있다고 알려져 있다.Korean red ginseng is a process by which unpeeled ginseng is steamed and then dried and aged repeatedly. Red ginseng has sedative and excitatory effects on the central nervous system, and acts on the circulatory system to prevent hypertension or arteriosclerosis. In addition, it lowers hematopoiesis and blood sugar, protects the liver, acts on the endocrine system, has an indirect effective action on sexual behavior and reproductive effects, has anti-inflammatory or anti-tumor action, and is known to have an effect of protecting and softening the skin. .
홍삼은 사포닌이라는 생리활성물질을 30여종이나 포함하고 있으며, 이 중 진세노사이드(ginsenoside) Rb1, Rb2, Rh1, Rh2, Rg2, Rg3, Rg5 및 Rf 등의 홍삼 특유의 14종 성분은 뛰어난 약리효과를 나타낸다고 알려져 있다. 사포닌은 카테콜아민(Cartecholamine)의 분비를 촉진시키며, 지질의 과산화 과정을 저해하는 항산화 활성을 지니며 배양세포에서의 돌연변이 및 세포변형에 대한 감소 효과 외에도 방사선 방어효과, 알킬화에 의한 염색체 이상 또는 미세핵의 방어효과가 있음이 보고된 바 있다. 또한, 홍삼은 사포닌 뿐 아닌 다당체, 탄수화물, 질소화합물, 폴리아세틸렌, 페놀산 및 비타민 등의 많은 생리활성물질을 포함하고 있다.Red ginseng contains about 30 kinds of physiologically active substances called saponins, and among them, 14 kinds of ingredients unique to red ginseng, such as ginsenoside Rb1, Rb2, Rh1, Rh2, Rg2, Rg3, Rg5 and Rf, have excellent pharmacological effects. is known to represent Saponin promotes the secretion of catecholamine, has antioxidant activity that inhibits the lipid peroxidation process, and has a radioactive protective effect, It has been reported to have a protective effect. In addition, red ginseng contains many physiologically active substances such as polysaccharides, carbohydrates, nitrogen compounds, polyacetylene, phenolic acid, and vitamins as well as saponins.
잘 먹고 잘사는 웰빙(well-being) 트렌드에 부응하여, 상기와 같이 보호, 탄력 및 지지역할을 하는 피부에 이용하기 위한 건강기능식품 및 화장품은 선택이 아닌 필수 요소로서 부가가치가 높은 산업으로 자리 잡아가고 있고, 고품질의 제품을 선호하는 현상이 두드러지고 있다.In response to the well-being trend of eating well and living well, health functional foods and cosmetics for use on the skin that act as protection, elasticity and support as described above are not an option, but an essential element, and are a high value-added industry. taking place and preference for high-quality products is prominent.
이에, 피부상태를 개선하기 위한 물질로서 레티노익산(retinoic acid), 동물 태반 유래의 단백질 등을 이용하여 콜라겐 합성을 촉진하고자 하는 시도가 있었으나, 레티노익산은 불안정하여 제형화하기 어려울 뿐만 아니라 안정성의 측면에서 제한이 있고, 동물 태반 유래 단백질은 광우병에 걸린 소의 적출물을 사용할 수 있다는 치명적인 단점이 있다.Accordingly, there have been attempts to promote collagen synthesis by using retinoic acid, animal placenta-derived protein, etc. as a material for improving the skin condition, but retinoic acid is unstable and difficult to formulate as well as in terms of stability However, there is a fatal disadvantage that animal placental-derived proteins can be used with extracts from cows with mad cow disease.
따라서, 본 발명자들은 피부 상태를 개선시킬 수 있는 물질을 발견하고자 많은 연구를 수행한 끝에, 정제어유로부터 획득한 오메가-7과 홍삼을 유효성분으로 함유하는 본 발명의 조성물이 우수한 피부상태 개선 효과를 가짐을 확인하고 본 발명을 완성하였다.Therefore, the present inventors have conducted many studies to discover substances that can improve skin condition, and the composition of the present invention containing omega-7 obtained from refined fish oil and red ginseng as active ingredients has an excellent skin condition improvement effect. It was confirmed that it has and completed the present invention.
본 발명의 목적은 정제어유 유래 오메가-7 및 홍삼을 포함하는 피부상태 개선용 조성물을 제공하는 것이다.An object of the present invention is to provide a composition for improving skin condition comprising omega-7 derived from refined fish oil and red ginseng.
상기 목적을 달성하기 위해, 본 발명은 정제어유 유래 오메가-7 및 홍삼을 포함하는 피부상태 개선용 건강기능식품 조성물을 제공한다.In order to achieve the above object, the present invention provides a health functional food composition for improving skin condition comprising omega-7 derived from refined fish oil and red ginseng.
또한, 본 발명은 정제어유 유래 오메가-7 및 홍삼을 포함하는 피부상태 개선용 식품 첨가물을 제공한다.In addition, the present invention provides a food additive for improving skin condition comprising omega-7 derived from refined fish oil and red ginseng.
또한, 본 발명은 정제어유 유래 오메가-7 및 홍삼을 포함하는 피부상태 개선용 화장료 조성물을 제공한다.In addition, the present invention provides a cosmetic composition for improving skin condition comprising omega-7 derived from refined fish oil and red ginseng.
본 발명에 따른 피부 상태 개선용 조성물은 정제어유 유래 오메가-7 및 사포닌, 다당체 등을 유효성분으로 함유하는 홍삼을 최적화된 농도로 포함하여, 높은 안정성을 기반으로 피부 보습도 개선, 피부보호 장벽 개선 및 산화적 스트레스에 의한 세포손상 저해 효과를 가져 미세먼지와 같은 외부 자극에 의한 피부의 손상에 대하여 종합적인 피부 상태 개선 효과를 발휘한다. 뿐만 아니라, 항산화, 항염증, 피부 탄력 및 주름 개선 효과를 가지고 있어 우수한 피부 노화 억제 효과를 나타낸다. 본 발명에 따른 조성물은 종합적으로 근본적인 피부 상태 개선 효과를 나타냄에 따라 식품, 건강기능식품, 식품 첨가물 및 화장품 등으로 다양하게 활용될 것으로 기대된다.The composition for improving skin condition according to the present invention contains red ginseng containing refined fish oil-derived omega-7 and saponins and polysaccharides as active ingredients in an optimized concentration, improving skin moisture and improving skin protection barrier based on high stability And it has an inhibitory effect on cell damage caused by oxidative stress, thereby exhibiting a comprehensive skin condition improvement effect against damage to the skin caused by external stimuli such as fine dust. In addition, it has antioxidant, anti-inflammatory, skin elasticity and anti-wrinkle effects, thereby showing excellent skin aging inhibitory effect. The composition according to the present invention is expected to be variously used as food, health functional food, food additive, cosmetics, etc. as it comprehensively exhibits a fundamental skin condition improvement effect.
도 1은 오메가-7과 홍삼 단독, 복합에서 세포 독성이 없음을 확인한 도이다.
도 2는 피부손상 개선 평가 시스템 및 HaCaT 세포(피부 각질형성세포)를 사용하여 오메가-7과 홍삼 단독, 복합에서 엘라스타제(Elastase) / 콜라게나제(Collagenase) 저해활성 및 MMP-1, MMP-3 생성능 확인 결과를 나타낸 도이다.
도 3은 오메가-7과 복합물 소재로 선정된 홍삼의 최적 배합비를 선정하기 위해 배합비를 달리한 각 실험군에서 엘라스타제(Elastase) / 콜라게나제(Collagenase) 저해활성 및 MMP-1, MMP-3 생성능을 확인한 결과를 나타낸 도이다.
도 4는 본 발명에 따른 조성물(오메가-7 및 홍삼을 3:7로 혼합)의 세포 독성을 확인한 도이다.
도 5는 DPPH 및 HaCaT 세포주에서의 ROS, SOD, GSH 변화량을 통하여 본 발명에 따른 조성물의 항산화 효과를 확인한 도이다. 구체적으로, 도 5a는 DPPH에 관한 결과, 도 5b는 ROS에 관한 결과, 도 5c는 SOD에 관한 결과, 도 5d는 GSH에 관한 결과를 나타낸 도이다.
도 6은 염증 사이토카인 발현 변화를 통하여 본 발명에 따른 조성물의 항염증 효과를 확인한 도이다. 구체적으로, ELISA 분석법을 통한 IL-1β(도 6a) 및 TNF-α(도 6b)의 농도 변화를 확인한 결과를 나타낸 도이다.
도 7은 염증성 인자 관련 단백질 발현 변화를 통하여 본 발명에 따른 조성물의 항염증 효과를 나타낸 도이다. 구체적으로, 웨스턴 블롯을 통해 COX-2(도 7a) 및 PGE2(도 7b)의 염증인자 발현량 변화를 확인한 결과를 나타낸 도이다.
도 8은 본 발명 조성물의 피부재생인자 활성 효과를 확인한 도이다. 구체적으로, 웨스턴 블롯 분석을 통한 MMP-1(도 8a), MMP-3(도 8b), PCOLA1(도 8c) 및 Elastin(도 8d)의 발현량 변화 확인 결과를 나타낸 도이다. 1 is a diagram confirming that there is no cytotoxicity in omega-7 and red ginseng alone or in combination.
Figure 2 shows Elastase / Collagenase inhibitory activity and MMP-1, MMP in omega-7 and red ginseng alone and in combination using a skin damage improvement evaluation system and HaCaT cells (skin keratinocytes) -3 It is a diagram showing the result of confirming the production ability.
3 is an Elastase / Collagenase inhibitory activity and MMP-1, MMP-3 in each experimental group in which the mixing ratio was changed to select the optimal mixing ratio of omega-7 and red ginseng selected as a composite material. It is a diagram showing the result of confirming the production ability.
Figure 4 is a diagram confirming the cytotoxicity of the composition (omega-7 and red ginseng 3:7 mixed) according to the present invention.
5 is a diagram confirming the antioxidant effect of the composition according to the present invention through the amount of change in ROS, SOD, and GSH in DPPH and HaCaT cell lines. Specifically, FIG. 5A is a diagram showing a result related to DPPH, FIG. 5B is a result related to ROS, FIG. 5C is a result related to SOD, and FIG. 5D is a result related to GSH.
6 is a view confirming the anti-inflammatory effect of the composition according to the present invention through the change in the expression of inflammatory cytokines. Specifically, it is a diagram showing the results of confirming the concentration change of IL-1β (Fig. 6a) and TNF-α (Fig. 6b) through the ELISA assay.
7 is a diagram showing the anti-inflammatory effect of the composition according to the present invention through changes in the expression of inflammatory factor-related proteins. Specifically, it is a diagram showing the result of confirming the expression level of the inflammatory factor of COX-2 (Fig. 7a) and PGE2 (Fig. 7b) through Western blot.
8 is a view confirming the skin regeneration factor activation effect of the composition of the present invention. Specifically, it is a diagram showing the result of confirming the expression level change of MMP-1 (Fig. 8a), MMP-3 (Fig. 8b), PCOLA1 (Fig. 8c) and Elastin (Fig. 8d) through Western blot analysis.
본 명세서 및 청구범위에 사용된 용어나 단어는 통상적이거나 사전적인 의미로 한정해서 해석되어서는 안 되며, 발명자는 그 자신의 발명을 가장 최선의 방법으로 설명하기 위해 용어의 개념을 적절하게 정의할 수 있다는 원칙에 입각하여 본 발명의 기술적 사상에 부합하는 의미와 개념으로 해석되어야만 한다.The terms or words used in the present specification and claims should not be construed as being limited to their ordinary or dictionary meanings, and the inventor may properly define the concept of the term in order to best describe his invention. It should be interpreted as meaning and concept consistent with the technical idea of the present invention based on the principle that there is.
본 발명은 정제어유 유래 오메가-7 및 홍삼을 포함하는 피부상태 개선용 조성물을 제공한다.The present invention provides a composition for improving skin condition comprising omega-7 derived from refined fish oil and red ginseng.
본 발명에 있어서, 용어 “오메가-7”은 사슬 말단에서 7번째 탄소에 이중결합 하나를 가지는 단일 불포화지방산으로 본 명세서에서 상기 오메가-7은 이에 제한되지는 않으나 정제어유로부터 수득한 것을 특징으로 한다.In the present invention, the term “omega-7” is a monounsaturated fatty acid having one double bond at the 7th carbon at the chain terminal. .
본 발명에 있어서, “정제어유”는 고도 불포화 지방산을 다량 함유하고 있는 어유(魚油, Fish oil)로 정제된 것을 의미하며, 상기 정제어유는 산자나무(sea buckthorn) 열매 오일에서 일반적으로 수득할 수 있는 것보다 오메가-7을 더 많은 양으로 포함하는 것이 특징이다. In the present invention, "refined fish oil" means a fish oil refined with a large amount of polyunsaturated fatty acids, and the refined fish oil can be generally obtained from sea buckthorn fruit oil. It is characterized by containing higher amounts of omega-7 than is present.
본 발명에 있어서, 상기 정제어유는 정제어유(생선기름)이면 제한 없이 사용할 수 있으나, 바람직하게는 상기 정제어유는 정제된 폴락 오일(refined pollock oil)일 수 있다. 본 발명의 일 실시예에서는 상기 정제어유로 7-MEGATM 500(알래스카 정제어유)을 사용하고 있으며, 상기 7-MEGATM 500의 주요 성분은 팔미톨레산(Palmitoleic acid, PLA)로 팔미톨레산이 총 중량의 50 내지 60 중량부를 차지하는 것을 특징으로 한다.In the present invention, the refined fish oil may be used without limitation as long as it is refined fish oil (fish oil), but preferably, the refined fish oil may be refined pollock oil. In one embodiment of the present invention, 7-MEGA TM 500 (Alaska refined fish oil) is used as the refined fish oil, and the main component of 7-MEGA TM 500 is palmitoleic acid (PLA), and palmitoleic acid is a total It is characterized in that it accounts for 50 to 60 parts by weight of the weight.
본 발명에 있어서, 상기 “오메가-7”은 오메가-7 함유 정제어유의 형태로 상기 조성물에 포함되는 것을 특징으로 한다. 상기 정제어유는 오메가-7 외에 올레산(Oleic acid, 오메가-9), 리놀렌산(α-Lenoleic acid, ALA, 오메가-3), DHA(오메가-3)와 같은 그 외 불포화 지방산을 더 포함하는 것을 특징으로 할 수 있다.In the present invention, the "omega-7" is characterized in that it is included in the composition in the form of omega-7-containing refined fish oil. The refined fish oil further comprises other unsaturated fatty acids such as oleic acid (omega-9), linoleic acid (α-Lenoleic acid, ALA, omega-3), and DHA (omega-3) in addition to omega-7 can be done with
본 발명은 상기 오메가-7 외에 복합 소재로서 홍삼을 더 포함하는 것을 특징으로 한다. 즉, 본 발명은 오메가-7 및 홍삼을 복합물로 포함하여 오메가-7 및 홍삼의 조합에 따른 시너지 효과로서의 우수한 피부 상태 개선 효과를 가지는 것을 특징으로 한다.The present invention is characterized in that it further comprises red ginseng as a composite material in addition to the omega-7. That is, the present invention is characterized in that it has an excellent skin condition improvement effect as a synergistic effect according to the combination of omega-7 and red ginseng by including omega-7 and red ginseng as a complex.
본 발명에 있어서, 상기 오메가-7 및 홍삼은 1:9 내지 9:1(w/w)의 중량비로 혼합될 수 있고, 바람직하게는 2:8 내지 8:2(w/w)의 중량비로 혼합될 수 있고, 보다 바람직하게는 2:8 내지 5:5(w/w)의 중량비로 혼합될 수 있고, 가장 바람직하게는 3:7(w/w)의 중량비로 혼합될 수 있으나, 이에 본 발명이 제한되는 것은 아니다. In the present invention, the omega-7 and red ginseng may be mixed in a weight ratio of 1:9 to 9:1 (w/w), and preferably in a weight ratio of 2:8 to 8:2 (w/w). may be mixed, more preferably in a weight ratio of 2:8 to 5:5 (w/w), and most preferably in a weight ratio of 3:7 (w/w), but this The present invention is not limited.
본 발명의 일 실시예에서는, 오메가-7 및 홍삼을 3:7(w/w) 중량비로 혼합하는 경우 엘라스타제(Elastase), 콜라게나제(Collagenase) 저해 활성이 다른 중량비로 혼합한 군에 비하여 현저히 높음을 확인하여, 오메가-7 및 홍삼을 혼합한 복합물의 시너지 효과로서 피부 개선 효과를 최대화할 수 있는 최적 비율을 확인하였다.In an embodiment of the present invention, when mixing omega-7 and red ginseng in a weight ratio of 3:7 (w/w), elastase and collagenase inhibitory activity are mixed in a different weight ratio. It was confirmed that it was significantly higher than that, and the optimal ratio that could maximize the skin improvement effect was confirmed as a synergistic effect of the complex mixed with omega-7 and red ginseng.
또한, 본 발명에 있어서, 상기 “홍삼”은 수삼을 정선하여 망상조직인 껍질을 벗기지 않은 채 증기로 쪄서 건조시킨 담황갈색 또는 담적갈색의 인삼을 의미하고, 수십 종의 진세노사이드로 분류되는 사포닌과 다당체 단백질을 유효성분으로 한다. 본 발명에 있어서 용어 “홍삼”은 상기 유효성분을 포함하는 홍삼 추출물, 홍삼 농축물 또는 홍삼 분말을 모두 포함하는 개념이다.In addition, in the present invention, the "red ginseng" refers to ginseng of light yellowish-brown or light reddish-brown color, which is selected from fresh ginseng and steamed with steam without peeling the reticulated tissue, and saponins classified into dozens of ginsenosides and Polysaccharide protein is used as an active ingredient. In the present invention, the term "red ginseng" is a concept including all of the red ginseng extract, red ginseng concentrate, or red ginseng powder containing the active ingredient.
본 발명에 있어서, 상기 홍삼 추출물은 물, C1 내지 C4의 저급알코올, 에틸아세테이트, 아세톤, 부틸아세테이트, 1,3-부틸렌 글리콜, 메틸렌클로라이드, 및 이들의 혼합용매로 이루어진 군으로부터 선택된 용매로 추출한 것일 수 있다. 상기 추출물은 이후 여과하거나 농축 또는 건조과정을 수행하여 용매를 제거할 수 있으며, 여과, 농축 및 건조를 모두 수행할 수 있다. 예컨대, 여과는 여과지를 이용하거나 감압여과기를 이용할 수 있으며, 농축은 감압 농축기, 건조는 동결건조법 등을 수행할 수 있으나, 이에 제한되는 것은 아니다.In the present invention, the red ginseng extract is extracted with a solvent selected from the group consisting of water, C1 to C4 lower alcohol, ethyl acetate, acetone, butyl acetate, 1,3-butylene glycol, methylene chloride, and a mixed solvent thereof. it could be The extract may then be filtered or concentrated or dried to remove the solvent, and both filtration, concentration and drying may be performed. For example, the filtration may be performed using filter paper or a reduced pressure filter, the concentration may be performed by a vacuum concentrator, and the drying may be performed by a freeze-drying method, but is not limited thereto.
또한, 상기 홍삼 농축물은 홍삼을 물이나 주정 또는 물과 주정을 혼합한 물질과 같은 용매로 추출·여과한 가용성 홍삼 성분을 농축한 것을 말한다. 상기 홍삼 농축물은 액체 상태 및 고체, 구체적으로 분말 상태를 모두 포함하는 개념이다. In addition, the red ginseng concentrate refers to a concentrated soluble red ginseng component obtained by extracting and filtering red ginseng with a solvent such as water, alcohol, or a mixture of water and alcohol. The red ginseng concentrate is a concept including both a liquid state and a solid state, specifically, a powder state.
홍삼은 별다른 부작용이 없으며 그 효과로는 항암, 노화억제, 항당뇨, 항스트레스, 뇌기능 강화, 빈혈 회복 효과, 피로 회복, 혈행 개선, 간기능 보호, 위장 기능 강화, 면역기능 증진 및 소염 작용 등이 알려져 있다(김낙두 외, 고려인삼학회지 20(4) 416-430, 1996, 정노팔 외, 고려인삼학회지 20(4) 431-471, 1996). 알려져 있는 홍삼의 주요 성분으로는 배당체(glycosides), 폴리아세틸렌계 화합물, 인삼향 성분(panacen), 함질소 성분, 항산화물질, 플라보노이드, 비타민 B군효소, 유기산, 아미노산 및 미량원소 등이 있다.Red ginseng has no side effects, and its effects include anti-cancer, anti-aging, anti-diabetes, anti-stress, brain function enhancement, anemia recovery effect, fatigue recovery, blood circulation improvement, liver function protection, gastrointestinal function enhancement, immune function enhancement and anti-inflammatory action, etc. is known (Nak-Doo Kim et al., Journal of the Korean Ginseng Society 20(4) 416-430, 1996, Nopal Jeong et al., Journal of the Korean Ginseng Society 20(4) 431-471, 1996). Known main ingredients of red ginseng include glycosides, polyacetylene-based compounds, ginseng flavoring ingredients (panacen), nitrogen-containing ingredients, antioxidants, flavonoids, vitamin B group enzymes, organic acids, amino acids, and trace elements.
본 발명에 있어서, 용어 “개선”은 조성물의 투여에 의해 피부상태가 개선, 보다 구체적으로 피부 보습도 개선, 피부보호 장벽 개선, 세포손상의 저해 또는 주름의 개수를 줄이거나 피부 표면이 보다 탄력성 있도록 하는 모든 행위를 의미한다.In the present invention, the term “improvement” means that the skin condition is improved by administration of the composition, more specifically, the skin moisture level is improved, the skin protective barrier is improved, the cell damage is inhibited or the number of wrinkles is reduced, or the skin surface is more elastic. means all actions
본 발명에 있어서, 용어 “피부상태 개선”은 피부 관능성 평가시 피부의 밝기, 투명도 등 피부 상태 전반에 대한 개선에 관한 것으로, 보다 구체적으로는 피부 자극에 의한 피부 손상 억제나 피부 표면 막 개선에 의한 것, 주름 개선 또는 피부 노화 억제에 의한 것을 모두 포함한다.In the present invention, the term "improving skin condition" relates to improvement of overall skin conditions such as brightness and transparency of the skin during skin sensory evaluation. It includes all of those caused by wrinkle improvement or skin aging inhibition.
본 발명에 있어서, 용어 “피부 자극에 의한 피부 손상 억제 또는 피부 표면막 개선”은 외부적 또는 내부적 요인에 의하여 피부의 진피를 포함한 전층에서 발생하는 피부 손상을 개선시키는 것을 말하며, 특히 피부의 표피로 나타내는 손상을 개선시키는 것을 말한다.In the present invention, the term “skin damage suppression or skin surface film improvement due to skin irritation” refers to improving skin damage occurring in the entire layer including the dermis of the skin due to external or internal factors, especially to the epidermis of the skin. refers to the improvement of the indicated damage.
본 발명에 있어서, 용어 “주름 개선 또는 피부 노화 억제”는 피부가 쇠하여 생긴 잔줄을 억제시키거나 이의 발생을 지연시키고, 피부가 보다 탄력적으로 보이도록 하는 모든 행위를 의미하는 것으로, 항산화 또는 다양한 요인들에 의한 MMP-1, 콜라겐 또는 엘라스틴의 분해를 억제함으로써 주름 개수 또는 피부 탄력성 증가 효과를 가지는 것을 특징으로 한다.In the present invention, the term "wrinkle improvement or skin aging inhibition" refers to any action that suppresses or delays the occurrence of fine lines caused by skin aging, and makes the skin look more elastic, antioxidant or various factors It is characterized in that it has the effect of increasing the number of wrinkles or skin elasticity by inhibiting the decomposition of MMP-1, collagen or elastin.
본 발명에 있어서, 용어 “피부 자극”은 물리적 자극 또는 화학적 자극인 것을 모두 포함하는 개념으로, 보다 구체적으로 상기 물리적 자극은 전기적 자극, 기계적 자극, 열적 자극, 빛, 소리 등을 포함하고, 화학적 자극은 산, 알칼리, 화학물질 등에 의한 자극을 포함한다. 이에 본 발명에 있어서 피부 자극은 바람직하게는 화학적 자극일 수 있으며, 보다 바람직하게는 미세먼지 등에 포함되는 화학물질에 의한 자극인 것일 수 있으나, 이에 제한되는 것은 아니다.In the present invention, the term “skin stimulation” is a concept that includes both physical stimulation and chemical stimulation, and more specifically, the physical stimulation includes electrical stimulation, mechanical stimulation, thermal stimulation, light, sound, etc., and chemical stimulation includes irritation by acids, alkalis, chemicals, etc. Accordingly, in the present invention, the skin irritation may be preferably chemical stimulation, and more preferably may be stimulation by chemicals contained in fine dust, etc., but is not limited thereto.
본 발명에 있어서, 상기 조성물은 식품 조성물, 건강기능식품 조성물 또는 식품 첨가물인 것을 특징으로 할 수 있다. 즉, 본 발명은 정제어유 유래 오메가-7 및 홍삼을 포함하는 피부 상태 개선용 식품 조성물, 정제어유 유래 오메가-7 및 홍삼을 포함하는 피부 상태 개선용 건강기능식품 조성물 및 정제어유 유래 오메가-7 및 홍삼을 포함하는 피부 상태 개선용 식품 첨가물을 제공한다. In the present invention, the composition may be characterized as a food composition, a health functional food composition or a food additive. That is, the present invention provides a food composition for improving skin condition comprising omega-7 and red ginseng derived from refined fish oil, a health functional food composition for improving skin condition comprising omega-7 derived from refined fish oil and red ginseng, and omega-7 derived from refined fish oil and It provides a food additive for improving skin condition, including red ginseng.
본 발명에 있어서, “식품”이란 인간이 먹기 위하여 요리하거나 또는 그대로 먹을 수 있는 모든 재료를 총칭하며, 이에 건강기능식품, 식품첨가물 등이 포함된다.In the present invention, "food" refers to all ingredients that can be cooked or eaten by humans, and includes health functional foods, food additives, and the like.
본 발명에 있어서, “건강기능식품”은 식품에 물리적, 생화학적, 생물공학적 수법 등을 이용하여 해당 식품의 기능을 특정 목적에 작용, 발현하도록 부가가치를 부여한 식품군이나 식품 조성이 갖는 생체방어리듬조절, 질병 방지와 회복 등에 관한 체내조절기능을 생체에 대하여 충분히 발현하도록 설계하여 가공한 식품으로, 장기적으로 복용하였을 때 인체에 무해하여야 한다.In the present invention, “health functional food” refers to a food group or food composition that has added value to act and express the function of the food for a specific purpose using physical, biochemical, and bioengineering methods, etc. It is a food that has been designed and processed to sufficiently express the body control functions related to disease prevention and recovery, etc., and should be harmless to the human body when taken for a long time.
본 발명에 있어서, “식품 첨가물”은 식품을 제조, 가공 또는 보존을 함에 있어 식품에 첨가, 혼합, 침윤, 기타의 방법으로 사용되는 물질에 관한 것으로, 건강기능식품과 같이 장기적으로 복용하였을 때 인체에 무해하여야 한다.In the present invention, "food additive" relates to a substance used in addition, mixing, infiltration, and other methods to food in manufacturing, processing or preserving food. should be harmless to
본 발명에 있어서, 상기 건강기능식품의 종류에 특별한 제한은 없다. 구체적으로, 본 발명의 조성물(오메가-7 + 홍삼)을 첨가할 수 있는 식품의 예로는 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제 등이 있으며, 특히 통상적인 의미에서의 해당 식품의 기능을 특정 목적에 작용, 발현하도록 부가가치를 부여한 식품군이나 식품 조성이 갖는 생체방어리듬조절, 질병 방지와 회복 등에 관한 체내조절기능을 생체에 대하여 충분히 발현하도록 설계된 식품을 모두 포함한다.In the present invention, there is no particular limitation on the type of the health functional food. Specifically, examples of foods to which the composition of the present invention (omega-7 + red ginseng) can be added include dairy products including ice cream, various soups, beverages, teas, drinks, alcoholic beverages, and vitamin complexes. All foods designed to sufficiently express the body control functions related to the biological defense rhythm control, disease prevention and recovery, etc. of the food group or food composition to which the added value is given to act and express the function of the food for a specific purpose in the sense of phosphorus. include
본 발명에 있어서, 상기 식품첨가물은 본 발명의 조성물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효성분의 혼합양은 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다. 일반적으로, 식품 또는 음료의 제조 시에 본 발명의 조성물은 원료에 대하여 15 중량% 이하, 바람직하게는 10 중량% 이하의 양으로 첨가된다. 그러나, 건강 및 위생을 목적으로 하거나 또는 건강 조절을 목적으로 하는 장기간의 섭취의 경우에는 상기 범위 이하일 수 있으며, 안전성 면에서 아무런 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로도 사용될 수 있다.In the present invention, the food additive may be added to the composition of the present invention as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method. The mixed amount of the active ingredient may be appropriately determined according to the purpose of use (prevention, health or therapeutic treatment). In general, in the production of food or beverage, the composition of the present invention is added in an amount of 15% by weight or less, preferably 10% by weight or less, based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health control, it may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.
본 발명에 있어서, 상기 조성물에는 식품학적으로 허용 가능한 다양한 식품 보조 첨가제를 포함할 수 있으며, 식품의 제조에 통상적으로 사용되는 적절한 담체, 부형제 및 희석제를 더욱 포함할 수 있다.In the present invention, the composition may include various food supplementary additives that are pharmaceutically acceptable, and may further include appropriate carriers, excipients and diluents commonly used in the manufacture of food.
보다 구체적으로 상기 외에 본 발명의 식품 조성물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 더 포함할 수 있다. 그 밖에 본 발명의 식품 조성물은 천연 과일주스, 과일주스 음료 및 야채 음료의 제조를 위한 과육을 포함할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 크게 중요하진 않지만 본 발명의 조성물 100 중량부 당 0.01 내지 0.1 중량부의 범위에서 선택되는 것이 일반적이다.More specifically, in addition to the above, the food composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, It may further include glycerin, alcohol, a carbonation agent used in carbonated beverages, and the like. In addition, the food composition of the present invention may include fruit for the production of natural fruit juice, fruit juice beverage, and vegetable beverage. These components may be used independently or in combination. The proportion of these additives is not very important, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.
또한, 본 발명에 따른 조성물은 화장료 조성물인 것을 특징으로 할 수 있다. 즉, 본 발명은 정제어유 유래 오메가-7 및 홍삼을 포함하는 피부 상태 개선용 화장료 조성물을 제공한다. In addition, the composition according to the present invention may be characterized as a cosmetic composition. That is, the present invention provides a cosmetic composition for improving skin condition comprising omega-7 derived from refined fish oil and red ginseng.
본 발명에 있어서, 용어 “화장료”는 인체의 외관을 아름답게 하는 모든 물건을 포함하는 개념이다.In the present invention, the term “cosmetics” is a concept that includes all things that make the appearance of the human body beautiful.
본 발명에 있어서, 상기 조성물은 피부상태 개선 효과를 갖는 공지의 유효성분을 1종 이상 더 포함할 수 있으며, 더욱이, 화장료 조성물에 통상적으로 사용되는 물질 1종 이상을 더 포함할 수 있다. 구체적으로 지방 물질, 유기 용매, 용해제, 농축제 및 겔화제, 연화제, 항산화제, 현탁제, 안정화제, 발포제(foaming agent), 방향제, 계면활성제, 물, 이온형 또는 비이온형 유화제, 충전제, 금속 이온 봉쇄제 및 킬레이트화제, 보존제, 비타민, 차단제, 습윤화제, 필수 오일, 염료, 안료, 친수성 또는 친유성 활성제, 지질 소낭 또는 화장품에 통상적으로 사용되는 임의의 다른 성분과 같은 화장품학 또는 피부 과학 분야 에서 통상적으로 사용되는 보조제를 추가로 함유할 수 있다. 또한, 상기의 성분들은 피부 과학 분야에서 일반적으로 사용되는 양으로 도입될 수 있다. In the present invention, the composition may further include one or more known active ingredients having a skin condition improvement effect, and further, may further include one or more substances commonly used in cosmetic compositions. Specifically, fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or non-ionic emulsifiers, fillers, in the field of cosmetology or dermatology, such as sequestering and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or any other ingredient commonly used in cosmetics. It may further contain adjuvants commonly used in In addition, the above ingredients may be introduced in an amount generally used in the field of dermatology.
본 발명에 있어서, 상기 조성물은 일반적인 유화 제형 및 가용화 제형의 형태로 제조할 수 있다. 유화 제형의 화장품으로는 영양화장수, 크림, 에센스 등이 있으며, 가용화 제형의 화장품으로는 유연화장수가 있다. 보다 구체적으로, 본 발명의 조성물은 용액, 겔, 고체 또는 반죽 무수 생성물, 수상에 유상을 분산시켜 얻은 에멀젼, 현탁액, 유탁액, 마이크로에멀젼, 마이크로캡슐, 마스크팩, 미세과립구 또는 이온형(리포좀), 비이온형의 소낭 분산 제의 형태, 크림, 스킨, 로션, 파우더, 연고, 스프레이, 페이스트, 팩, 세안제, 비누, 계면활성제 함유 클린싱, 오일, 분말 파운데이션, 목욕용 파우더, 유탁액 파운데이션, 왁스, 파운데이션 또는 콘실 스틱의 형태로 제공될 수 있다. 또한, 포말 (foam)의 형태 또는 압축된 추진제를 더 함유한 에어로졸 조성물의 형태로도 제조될 수 있다.In the present invention, the composition can be prepared in the form of a general emulsified formulation and a solubilized formulation. Cosmetics with emulsified formulations include nutritional lotions, creams, and essences, and solubilized cosmetics include softened lotions. More specifically, the composition of the present invention is a solution, a gel, a solid or kneaded anhydrous product, an emulsion obtained by dispersing an oil phase in an aqueous phase, a suspension, an emulsion, a microemulsion, a microcapsule, a mask pack, a microgranule or an ionic (liposome) , Non-ionic form of vesicle dispersant, cream, skin, lotion, powder, ointment, spray, paste, pack, face wash, soap, cleansing with surfactant, oil, powder foundation, bath powder, emulsion foundation, wax, It may be provided in the form of a foundation or a concealer stick. In addition, it may be prepared in the form of a foam or an aerosol composition further containing a compressed propellant.
상기 조성물의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로 동물성유, 식물성유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크, 산화아연 등이 포함될 수 있다. When the formulation of the composition is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. may be included as carrier components. there is.
상기 조성물의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로 락토스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트, 폴리아미드 파우더 등이 포함될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄, 디메틸 에테르 등의 추진체를 포함할 수 있다. When the formulation of the composition is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, etc. may be included as a carrier component. In particular, in the case of a spray, additional chlorofluorohydrocarbon, propane / May contain propellants such as butane and dimethyl ether.
상기 조성물의 제형이 용액 또는 유탁액인 경우에는 담체 성분으로 용매, 용해화제, 유탁화제 등이 포함될 수 있고, 구체적으로 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프 로필렌글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜, 소르비탄의 지방산 에스테 르 등이 포함될 수 있다. When the formulation of the composition is a solution or emulsion, a solvent, a solubilizer, an emulsifier, etc. may be included as carrier components, and specifically, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Lopylene glycol, 1,3-butyl glycol oil, glycerol fatty esters, polyethylene glycol, fatty acid esters of sorbitan, and the like may be included.
상기 조성물의 제형이 현탁액인 경우에는 담체 성분으로 물, 에탄올, 프로필렌글리콜 등의 액상 희석제; 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르, 폴리옥시에틸렌 소르비탄 에스테르 등의 현탁제; 미 소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가, 트라칸트 등이 포함될 수 있다. When the formulation of the composition is a suspension, as a carrier component, a liquid diluent such as water, ethanol, propylene glycol; suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester; Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, tracanth, and the like may be included.
상기 조성물의 제형이 계면활성제 함유 클린징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르 설페이트, 설포숙신산 모노에스테르, 이세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사르코시 네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산 디에탄 올아미드, 식물성유, 라놀린유도체, 에톡실화 글리세롤 지방산 에스테르 등이 포함될 수 있다.When the formulation of the composition is surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether as carrier components Sulfates, alkylamidobetaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, ethoxylated glycerol fatty acid esters, and the like may be included.
또한, 이외에 첨가해도 되는 배합 성분은 상기 예시로 제한되는 것은 아니며, 상기 어느 성분도 본 발명의 목적 및 효과를 손상시키지 않는 범위 내에서 배합 가능하지만, 총 중량에 대하여 바람직하게는 0.01 내지 10% 중량 백분율, 보다 바람직하게는 0.01 내지 5% 중량 백분율로 배합된다. In addition, the blending components that may be added are not limited to the above examples, and any of the above ingredients can be blended within a range that does not impair the object and effect of the present invention, but preferably 0.01 to 10% by weight based on the total weight. , more preferably 0.01 to 5% by weight.
중복되는 내용은 본 명세서의 복잡성을 고려하여 생락하며, 본 명세서에서 달리 정의되지 않은 용어들은 본 발명이 속하는 기술분야에서 통상적으로 사용되는 의미를 갖는 것이다.Redundant content is omitted in consideration of the complexity of the present specification, and terms not defined otherwise in the present specification have the meanings commonly used in the art to which the present invention pertains.
이하, 본 발명의 이해를 돕기 위하여 실시예, 실험예 및 제조예를 들어 상세하게 설명하기로 한다. 다만 하기의 실시예, 실험예 및 제조예는 본 발명의 내용을 예시하는 것일 뿐 본 발명의 범위가 하기 실시예, 실험예 및 제조예에 한정되는 것은 아니다. 본 발명의 실시예, 실험예 및 제조예는 당업계에서 평균적인 지식을 가진 자에게 본 발명을 보다 완전하게 설명하기 위해 제공되는 것이다.Hereinafter, Examples, Experimental Examples, and Preparation Examples will be described in detail to help the understanding of the present invention. However, the following Examples, Experimental Examples and Preparation Examples are merely illustrative of the contents of the present invention, and the scope of the present invention is not limited to the following Examples, Experimental Examples and Preparation Examples. The Examples, Experimental Examples and Preparation Examples of the present invention are provided to more completely explain the present invention to those of ordinary skill in the art.
실시예 1. 오메가-7과의 최적 복합물 도출Example 1. Derivation of optimal complexes with omega-7
오메가-7과의 최적 복합물을 도출하기 위하여 복합 소재로 홍삼을 선정하고 피부 개선 효과를 평가하였다. 오메가-7은 7-MEGATM 500(알래스카 정제어유; Organic technologies, 미국)을 사용하고 홍삼은 멸균증류수로 희석 후에 EtOH(94.5%)과 섞은 홍삼 농축물(한국인삼공사)을 사용하여 5:5 비율로 혼합하였다. 한편, 상기 7-MEGATM 500의 구체적인 구성성분 및 각 구성성분의 함량을 하기 표 1에 나타내었다.To derive the optimal complex with omega-7, red ginseng was selected as a composite material and the skin improvement effect was evaluated. Omega-7 uses 7-MEGA TM 500 (Alaskan refined fish oil; Organic technologies, USA), and red ginseng is diluted with sterile distilled water and then mixed with EtOH (94.5%) using red ginseng concentrate (Korea Ginseng Corporation) 5:5 mixed in proportions. Meanwhile, the specific components of the 7-
[표 1][Table 1]
최종 용적에 맞추어 용매를 채워준 뒤, 실린지 필터를 사용하여 여과 후 실험에 사용하였다. 대조군으로는 단독 오메가-7 또는 단독 홍삼을 동량으로 사용하였다.After filling the solvent according to the final volume, it was used for the experiment after filtration using a syringe filter. As a control group, single omega-7 or single red ginseng was used in the same amount.
실험예 1. 세포 독성 확인Experimental Example 1. Confirmation of cytotoxicity
XTT assay kit(Roche, Indianapolis, IN, USA) 분석을 통하여 각질형성세포인 HaCaT 세포주의 증식효과를 확인하여 오메가-7 및 홍삼의 세포 독성을 확인하였다. 상기 HaCaT 세포주는 American Type Culture Collection(ATCC, USA)에서 구매하여 실험을 진행하였고, 세포는 DMEM 배지, 10% 소태아혈청, 50 U/ml P/S (페니실린/스트렙토마이신)에서 37℃, 습도 95%, 5%의 CO2, CO2 인큐베이터(Sanyo, Japan) 조건 하에서 배양한 세포주를 실험에 사용하였다.Through the XTT assay kit (Roche, Indianapolis, IN, USA) analysis, the proliferation effect of the HaCaT cell line, a keratinocyte, was confirmed to confirm the cytotoxicity of omega-7 and red ginseng. The HaCaT cell line was purchased from the American Type Culture Collection (ATCC, USA) and the experiment was performed, and the cells were cultured in DMEM medium, 10% fetal bovine serum, 50 U/ml P/S (penicillin/streptomycin) at 37°C, humidity. A cell line cultured under 95%, 5% CO 2 , CO 2 incubator conditions (Sanyo, Japan) was used for the experiment.
먼저, HaCaT 세포를 96-웰 플레이트에 1×104 세포/웰이 되도록 분주하고 24시간 동안 안정화한 후, 상기 실시예 1의 조성물을 100 μg/ml 농도로 처리하고 24시간 동안 배양하였다. 배양이 끝난 후 50 ㎕의 XTT 용액을 각각의 웰에 첨가하고 4시간 배양 후 ELISA reader(Molecular Devices, CA, USA)를 이용하여 450 nm에서 흡광도를 측정하여 세포 사멸 정도를 분석하였다. 그 결과는 도 1에 나타내었다.First, HaCaT cells were aliquoted in a 96-well plate to 1×10 4 cells/well and stabilized for 24 hours, then the composition of Example 1 was treated at a concentration of 100 μg/ml and cultured for 24 hours. After incubation, 50 μl of XTT solution was added to each well, and after 4 hours of incubation, the degree of cell death was analyzed by measuring absorbance at 450 nm using an ELISA reader (Molecular Devices, CA, USA). The results are shown in FIG. 1 .
도 1에 나타낸 바와 같이, 오메가-7 및 홍삼 복합물(5:5)을 처리한 경우, 피부각질세포인 HaCaT 세포주에서 세포 독성이 나타나지 않음을 확인하였다. As shown in FIG. 1 , when the omega-7 and red ginseng complex (5:5) was treated, it was confirmed that cytotoxicity did not appear in the HaCaT cell line, which is keratinocytes of the skin.
실험예 2. 엘라스타제, 콜라게나제 저해활성 및 MMP-1, 3 활성 확인Experimental Example 2. Elastase, collagenase inhibitory activity and MMP-1, 3 activity confirmation
2-1. 엘라스타제(Elastase) 저해활성 2-1. Elastase inhibitory activity
엘라스타제 저해활성 측정은 0.1 M Tris-HCl buffer (pH 8.0) 용액 100 ㎕에 2mM N-succinyl-(Ala)3-p-nitroaniline을 20 ㎕씩 분주하고, 각 시료를 20 ㎕씩 가하여 수행하였다. 여기에 0.2U 엘라스타제 효소 20 ㎕를 분주한 후 반응액을 37℃에서 30분 동안 배양하여 410 nm에서 흡광도(Molecular Devices, CA, USA)를 측정하였다. 양성 대조군으로는 100 mg/ml/의 아스코르브산을 사용하였으며, 대조군은 시료 대신 버퍼 용액을 사용하였다. 저해활성도 계산은 inhibitory activity (%) = [1 - (sample-blank)] X 100으로 계산하였다.Elastase inhibitory activity was measured by dispensing 20 μl of 2mM N-succinyl-(Ala)3-p-nitroaniline into 100 μl of 0.1 M Tris-HCl buffer (pH 8.0), and adding 20 μl of each sample. . After dispensing 20 μl of 0.2U elastase enzyme, the reaction solution was incubated at 37° C. for 30 minutes to measure absorbance at 410 nm (Molecular Devices, CA, USA). As a positive control, 100 mg/ml/ascorbic acid was used, and as a control, a buffer solution was used instead of the sample. Inhibitory activity was calculated as inhibitory activity (%) = [1 - (sample-blank)]
그 결과, 도 2(상단 패널)에 나타낸 바와 같이, 오메가-7 및 홍삼 복합물(5:5)은 오메가-7 또는 홍삼 단독 처리시와 비교하여 엘라스타제 저해 활성이 현저히 우수함을 확인하였다.As a result, as shown in FIG. 2 (upper panel), it was confirmed that the omega-7 and red ginseng complex (5:5) had significantly superior elastase inhibitory activity compared to the omega-7 or red ginseng alone treatment.
2-2. 콜라게나제(Collagenase) 저해활성 2-2. Collagenase inhibitory activity
0.1M Tris-HCl 버퍼(pH 7.5)에 4 mM CaCl2를 첨가하고, 4-phenylazoben zyloxycarbonyl-Pro-Leu-Gly-Pro-D-Arg(0.3 mg/ml)를 녹인 기질액 250 ㎕ 및 시료용액 100 ㎕의 혼합액에 콜라게나제(collagenase)(0.2 mg/ml) 150 ㎕를 첨가하여 실온에서 20분 간 방치한 후 6% 시트르산 500 ㎕을 넣어 반응을 정지시켰다. 그 후, 에틸 아세테이트 1.5 ml를 첨가하여 320 nm에서 형광분광광도계(BioSpectrometer fluorescence, Eppendorf)를 사용하여 측정하였다. 저해활성도 계산은 inhibitory activity (%) = [1 - (sample-blank)] X 100으로 계산하였다. 250 μl of substrate solution and sample solution in which 4 mM CaCl 2 was added to 0.1M Tris-HCl buffer (pH 7.5) and 4-phenylazoben zyloxycarbonyl-Pro-Leu-Gly-Pro-D-Arg (0.3 mg/ml) was dissolved 150 μl of collagenase (0.2 mg/ml) was added to 100 μl of the mixed solution, left at room temperature for 20 minutes, and then 500 μl of 6% citric acid was added to stop the reaction. Then, 1.5 ml of ethyl acetate was added, and measurement was performed using a fluorescence spectrophotometer (BioSpectrometer fluorescence, Eppendorf) at 320 nm. Inhibitory activity was calculated as inhibitory activity (%) = [1 - (sample-blank)]
그 결과, 도 2(상단 패널)에 나타낸 바와 같이, 오메가-7 및 홍삼 복합물(5:5)은 오메가-7 또는 홍삼 단독 처리시와 비교하여 콜라게나제 저해 활성이 현저히 우수함을 확인하였다.As a result, as shown in FIG. 2 (upper panel), it was confirmed that the omega-7 and red ginseng complex (5:5) had significantly superior collagenase inhibitory activity compared to the omega-7 or red ginseng alone treatment.
2-3. MMP-1과 MMP-3 활성 2-3. MMP-1 and MMP-3 activity
MMP(matrix metalloproteinase)는 세포외기질(extracellular matrix)과 기저막 분해에 관여하는 효소군으로써, MMP-1 또는 MMP-3은 콜라겐 분해와 관여하여 가장 중요하게 작용하는 것으로 알려져 있다. 따라서, MMP-1, MMP-3 발현 억제 분석을 통하여 본 발명에 따른 복합물의 주름개선 효과를 확인하고자 하였다. 구체적으로, HaCaT세포의 배양액에서 분비되는 MMP-1과 MMP-3의 양을 R&D system에서 구입한 ELISA kit을 이용하여 측정하였다. HaCaT 세포 배양액은 세포를 1×104 cells/웰의 밀도로 96-웰 플레이트에 분주하고 각 시료들을 처리하고 24시간 동안 배양한 후에 얻어진 것으로 배양액 100 ㎕를 넣고 실온에 2시간, 컨쥬게이트(conjugate)를 200 ㎕ 넣고 실온에서 2시간, 기질 용액(substrate solution) 200 ㎕를 넣고 실온에서 20분간 방치하였다. 각각의 과정 사이에 세척을 수행하였으며 마지막 단계에서는 세척하지 않은 채로 50 ㎕의 정지 용액(stop solution)을 넣고 450 nm에서 흡광도 (Molecular Devices, CA, USA)를 측정하였다.MMP (matrix metalloproteinase) is a group of enzymes involved in the decomposition of the extracellular matrix and basement membrane, and MMP-1 or MMP-3 is known to be the most important because it is involved in collagen degradation. Therefore, it was attempted to confirm the anti-wrinkle effect of the composite according to the present invention through the MMP-1 and MMP-3 expression inhibition analysis. Specifically, the amounts of MMP-1 and MMP-3 secreted from the culture medium of HaCaT cells were measured using an ELISA kit purchased from the R&D system. HaCaT cell culture solution was obtained after dispensing cells in a 96-well plate at a density of 1×10 4 cells/well, processing each sample, and culturing for 24 hours. ) was added, 200 μl of the substrate solution was added at room temperature for 2 hours, and 200 μl of the substrate solution was added and left at room temperature for 20 minutes. Washing was performed between each process, and in the last step, 50 μl of a stop solution was added without washing, and absorbance at 450 nm (Molecular Devices, CA, USA) was measured.
그 결과, 도 2(하단 패널)에 나타낸 바와 같이, 오메가-7 및 홍삼 복합물(5:5)은 오메가-7 또는 홍삼 단독 처리시와 비교하여 MMP-1 및 MMP-3의 발현 저해 효과가 우수함을 확인하였다. 상기 결과를 통해, 오메가-7 및 홍삼 복합물은 내인성 또는 광노화에 의해 발생하는 피부조직 분해효소인 MMP-1 및 MMP-3의 생합성을 저해하여 피부노화를 억제하고 피부 주름 개선에 효과적임을 확인하였다.As a result, as shown in FIG. 2 (lower panel), the omega-7 and red ginseng complex (5:5) has an excellent effect on inhibiting the expression of MMP-1 and MMP-3 compared to the omega-7 or red ginseng alone treatment. was confirmed. Through the above results, it was confirmed that the omega-7 and red ginseng complex inhibited the biosynthesis of MMP-1 and MMP-3, which are skin tissue degrading enzymes caused by endogenous or photoaging, to inhibit skin aging and to be effective in improving skin wrinkles.
실시예 2. 본 발명에 따른 오메가-7 및 홍삼 복합 조성물의 제조Example 2. Preparation of omega-7 and red ginseng complex composition according to the present invention
상기 실시예 1 및 실험예 1 내지 2를 통해 홍삼이 오메가-7과의 최적 복합 소재가 될 수 있음을 확인하였다. 이에, 이번에는 실험을 통해 오메가-7과 홍삼의 최적 배합비를 도출하고자 하였다. 오메가-7과 홍삼의 최적 배합비를 도출하기 위하여 오메가-7 및 홍삼을 1:9 내지 9:1로 중량비를 달리하여 혼합하고(1:9, 3:7, 5:5, 7:3, 9:1), 각 혼합물에 대하여 엘라스타제, 콜라게나제 저해활성 및 MMP-1, MMP-3 저해활성을 측정하였다. 실험은 상기 실험예 2와 동일한 방법을 사용하여 수행하였다.Through Example 1 and Experimental Examples 1 and 2, it was confirmed that red ginseng could be an optimal composite material with omega-7. Therefore, this time, we tried to derive the optimal mixing ratio of omega-7 and red ginseng through an experiment. To derive the optimal mixing ratio of omega-7 and red ginseng, omega-7 and red ginseng are mixed with different weight ratios from 1:9 to 9:1 (1:9, 3:7, 5:5, 7:3, 9 : 1), elastase, collagenase inhibitory activity and MMP-1, MMP-3 inhibitory activity were measured for each mixture. The experiment was performed using the same method as in Experimental Example 2.
그 결과, 도 3에 나타낸 바와 같이, 모든 실험군에서 오메가-7 및 홍삼 혼합물은 엘라스타제, 콜라게나제 저해 활성 및 MMP-1, 3 활성이 우수한 것으로 나타났으며, 특히, 오메가-7:홍삼을 3:7 중량비로 혼합한 실험군에서 다른 실험군에 비해 저해활성이 현저히 높게 나타남을 확인하여 오메가-7:홍삼을 3:7로 혼합하는 것이 최적의 배합비임을 확인하였다. 이에, 이후 실험에서는 3:7로 혼합한 오메가-7 및 홍삼 복합물(이하, 본 발명의 조성물)을 사용하여 실험을 진행하였다.As a result, as shown in FIG. 3, the omega-7 and red ginseng mixture in all experimental groups showed excellent elastase, collagenase inhibitory activity and MMP-1, 3 activity, in particular, omega-7: red ginseng. It was confirmed that the inhibitory activity was significantly higher than that of the other experimental groups in the experimental group in which omega-7: red ginseng was mixed at 3:7 by weight in the experimental group mixed with omega-7: the optimal mixing ratio. Therefore, in the subsequent experiment, an experiment was conducted using a 3:7 mixture of omega-7 and red ginseng complex (hereinafter, the composition of the present invention).
실험예 3. 세포 독성 확인Experimental Example 3. Confirmation of cytotoxicity
오메가-7 및 홍삼을 3:7로 혼합한 본 발명의 조성물이 HaCaT 세포의 세포 생존률에 미치는 영향을 MTT assay 방법을 응용하여 측정하였다. HaCaT 세포에서의 증식 효과는 HaCaT 세포를 48-웰 플레이트에 2x105 cells/웰이 되도록 분주하고 시료를 각각 농도별로(5, 10, 50, 100 μg/ml) 처리한 뒤 control(CON)을 제외한 나머지 처리군에 H2O2를 처리하고 24시간 배양하였다. 그 후에 250 μg/ml MTT 용액을 첨가하여 4시간 동안 배양한 후 배양액을 제거하고 200 ㎕의 dimethyl sulfoxide를 가하여 10분간 실온에서 반응 후 570 nm에서 흡광도를 측정하였다.The effect of the composition of the present invention in which omega-7 and red ginseng were mixed at 3:7 on the cell viability of HaCaT cells was measured by applying the MTT assay method. The proliferative effect in HaCaT cells was obtained by dispensing HaCaT cells to 2x10 5 cells/well in a 48-well plate and treating each sample by concentration (5, 10, 50, 100 μg/ml), except for control (CON). The remaining treatment groups were treated with H 2 O 2 and cultured for 24 hours. After that, 250 μg/ml MTT solution was added and incubated for 4 hours, the culture medium was removed, 200 μl of dimethyl sulfoxide was added, reacted at room temperature for 10 minutes, and absorbance was measured at 570 nm.
그 결과, 도 4에 나타낸 바와 같이, 본 발명의 조성물(오메가-7 및 홍삼의 3:7 복합물) 처리 농도 증가에 따라 농도 의존적으로 세포생존율이 증가함을 확인하였고, 100 μg/ml 처리 농도에서 H2O2 단독처리군 대비 세포생존율이 33% 증가됨을 확인하였다. 동일농도(100 μg/ml)의 홍삼 단독처리군과 비교할 때에는 세포생존율이 28% 증가함을 확인하였다. 상기 결과를 통해, 본 발명의 조성물은 산화적 스트레스에 의한 세포손상 저해 효과가 있고, 그 효과는 홍삼보다 뛰어남을 확인하였다.As a result, as shown in FIG. 4, it was confirmed that the cell viability increased in a concentration-dependent manner according to the increase in the treatment concentration of the composition of the present invention (3:7 complex of omega-7 and red ginseng), and at 100 μg/ml treatment concentration It was confirmed that the cell viability was increased by 33% compared to the H 2 O 2 single treatment group. When compared with the group treated with red ginseng alone at the same concentration (100 μg/ml), it was confirmed that the cell viability increased by 28%. Through the above results, it was confirmed that the composition of the present invention has an inhibitory effect on cell damage caused by oxidative stress, and the effect is superior to that of red ginseng.
실험예 4. 본 발명 조성물의 항산화 효과 확인Experimental Example 4. Confirmation of the antioxidant effect of the composition of the present invention
항산화 관련 인자, 즉 DPPH 자유 라디칼 소거능 및 HaCaT 세포주에서의 ROS, SOD, GSH의 농도 변화를 통한 본 발명 조성물의 우수한 항산화 효과를 확인하였다. 상기 HaCaT 세포주는 American Type Culture Collection(ATCC, USA)에서 구매하여 실험을 진행하였고, 세포는 DMEM 배지, 10% 소태아혈청, 50 U/ml P/S (페니실린/스트렙토마이신)에서 37℃, 습도 95%, 5%의 CO2, CO2 인큐베이터(Sanyo, Japan) 조건 하에서 배양한 세포주를 실험에 사용하였다. The excellent antioxidant effect of the composition of the present invention was confirmed by changing the concentration of antioxidant-related factors, namely, DPPH free radical scavenging ability and ROS, SOD, and GSH in HaCaT cell lines. The HaCaT cell line was purchased from the American Type Culture Collection (ATCC, USA) and the experiment was performed, and the cells were cultured in DMEM medium, 10% fetal bovine serum, 50 U/ml P/S (penicillin/streptomycin) at 37°C, humidity. A cell line cultured under 95%, 5% CO 2 , CO 2 incubator conditions (Sanyo, Japan) was used for the experiment.
4-1. DPPH(α, α-diphenyl-β-picrylhydrazyl) 자유라디칼 소거능4-1. DPPH (α, α-diphenyl-β-picrylhydrazyl) free radical scavenging activity
대표적인 항산화 지표로 널리 알려진 아스코르브산(비타민C, VitC, Asc)을 양성 대조군으로 하여 본 발명의 조성물의 농도를 1 mg/mL로 맞추었다. 이 후 상기 실험군 10 ㎕와 DPPH 용액(126 μg/mL) 90 ㎕를 섞은 후 상온에서 10분간 방치하여 배양하였으며, 517 nm에서 흡광도를 측정하였다.Ascorbic acid (vitamin C, VitC, Asc), which is widely known as a representative antioxidant index, was used as a positive control to adjust the concentration of the composition of the present invention to 1 mg/mL. After that, 10 μl of the experimental group and 90 μl of DPPH solution (126 μg/mL) were mixed and incubated at room temperature for 10 minutes, and absorbance was measured at 517 nm.
그 결과, 도 5a에 나타낸 바와 같이, 본 발명의 조성물은 아스코르브산 대비 46.83±2.19%의 DPPH 자유 라디칼 소거능을 가짐을 확인하였다. 또한, 동일농도(100 μg/ml)의 홍삼 단독처리군과 비교할 때에는 DPPH 자유라디칼 소거능이 33% 증가하는바, 본 발명의 복합처리군의 효과가 홍삼 단독처리군보다 뛰어남을 확인하였다.As a result, as shown in Figure 5a, it was confirmed that the composition of the present invention has a DPPH free radical scavenging ability of 46.83±2.19% compared to ascorbic acid. In addition, the DPPH free radical scavenging ability increased by 33% when compared with the red ginseng alone treatment group at the same concentration (100 μg/ml), confirming that the effect of the combined treatment group of the present invention was superior to that of the red ginseng alone treatment group.
4-2. ROS(reactive oxygen species) 저해 활성4-2. ROS (reactive oxygen species) inhibitory activity
HaCaT 세포를 48-웰 플레이트에 2×105 세포/웰이 되도록 분주하였다. 48시간 동안 배양한 후, 본 발명의 조성물을 처리하여 1시간 동안 배양하였고, 이 후 정상대조군(Vehicle)을 제외한 나머지 실험군에 1 mM H2O2를 처리하여 10분 동안 배양하였다. 이후, PBS와 DCFDA(2′,7′-Dichlorofluorescin diacetate)를 5000:1로 희석시킨 용액을 배지와 동량으로 처리하여 20분 동안 반응시킨 후 형광발광량을 측정하였다.HaCaT cells were seeded in a 48-well plate at a concentration of 2×10 5 cells/well. After culturing for 48 hours, the composition of the present invention was treated and cultured for 1 hour, and then the remaining experimental groups except for the normal control group (Vehicle) were treated with 1 mM H 2 O 2 and incubated for 10 minutes. Thereafter, a solution of PBS and DCFDA (2′,7′-Dichlorofluorescin diacetate) diluted at 5000:1 was treated with the same amount as the medium, reacted for 20 minutes, and then the amount of fluorescence was measured.
그 결과, 도 5b에 나타낸 바와 같이, 본 발명의 조성물 처리 농도 증가에 따라 농도 의존적으로 ROS 생성이 감소되는 경향을 보이며, 100 μg/ml 농도에서 H2O2 단독처리군 대비 ROS 발현이 34% 감소함을 확인하였다. 동일농도(100 μg/ml)의 홍삼 단독처리군과 비교할 때에는 ROS 감소능이 24% 증가함을 확인하였다. 상기 결과를 통해, 산화적 스트레스로 인하여 증가하는 것으로 알려진 ROS의 농도가 본 발명의 조성물의 처리에 의하여 농도 의존적으로 유의성 있게 감소하며, 그 효과는 홍삼 단독처리군보다 뛰어남을 확인하였다.As a result, as shown in FIG. 5b , ROS production tends to decrease in a concentration-dependent manner as the concentration of the composition of the present invention increases, and at a concentration of 100 μg/ml, ROS expression is 34% compared to the H 2 O 2 alone treatment group. decrease was confirmed. It was confirmed that the ROS reduction ability was increased by 24% when compared with the group treated with red ginseng alone at the same concentration (100 μg/ml). Through the above results, it was confirmed that the concentration of ROS, which is known to increase due to oxidative stress, was significantly decreased in a concentration-dependent manner by the treatment of the composition of the present invention, and the effect was superior to that of the red ginseng alone treatment group.
4-3. SOD(Superoxide Dismutase) 증가 활성4-3. SOD (Superoxide Dismutase) increase activity
HaCaT 세포를 세포배양용 플레이트(SPL사, 100 cm2)에 5×106 세포/ml가 되도록 분주하였다. 48시간 동안 배양한 후, 본 발명의 조성물을 처리하였고, 이 후 정상대조군(Vehicle)을 제외한 나머지 실험군에 1 mM H2O2를 처리한 후 48시간 동안 배양하였다. 그 후 세포 분쇄액이 희석된 완충액을 샘플로 하여 샘플 10 ㎕에 radical detector 200 ㎕, 잔틴 산화효소(Xanthine oxidase) 20 ㎕를 넣고 상온에서 30분 동안 반응시킨 후, 440~460 nm 범위에서 흡광도를 측정하였다.HaCaT cells were aliquoted to a cell culture plate (SPL, 100 cm 2 ) at a concentration of 5×10 6 cells/ml. After culturing for 48 hours, the composition of the present invention was treated, and thereafter, 1 mM H 2 O 2 was treated in the remaining experimental groups except for the normal control group (Vehicle), and then cultured for 48 hours. After that, using the buffer solution diluted with the cell disruption solution as a sample, 200 μl of radical detector and 20 μl of xanthine oxidase were added to 10 μl of the sample and reacted at room temperature for 30 minutes, and absorbance was measured in the range of 440 to 460 nm. measured.
그 결과, 도 5c에 나타낸 바와 같이, 본 발명의 조성물 처리 농도 증가에 따라 농도 의존적으로 SOD 발현이 증가하는 경향을 보이며, 100 μg/ml 농도에서 H2O2 단독처리군 대비 SOD 발현이 29% 증가됨을 확인하였다. 동일농도(100 μg/ml)의 홍삼 단독처리군과 비교할 때에는 SOD 발현이 26% 증가함을 확인하였다. 상기 결과를 통해, 항산화 방어기작으로 발현이 증가하는 SOD가 본 발명의 조성물에 의하여 증가하며, 그 효과는 홍삼 단독처리군보다 뛰어남을 확인하였다.As a result, as shown in FIG. 5c , the SOD expression tends to increase in a concentration-dependent manner according to the increase in the treatment concentration of the composition of the present invention, and at a concentration of 100 μg/ml, the SOD expression is 29% compared to the H 2 O 2 alone treatment group. was confirmed to be increased. It was confirmed that the expression of SOD was increased by 26% when compared with the group treated with red ginseng alone at the same concentration (100 μg/ml). Through the above results, it was confirmed that the SOD, the expression of which is increased as an antioxidant defense mechanism, is increased by the composition of the present invention, and the effect is superior to that of the red ginseng alone treatment group.
4-4. GSH(Glutathione) 증가 활성4-4. GSH (Glutathione) increasing activity
HaCaT 세포를 세포배양용 플레이트(SPL사, 100 cm2)에 5×106 세포/ml이 되도록 분주하였다. 48시간 동안 배양한 후, 본 발명의 조성물을 처리하였고, 이 후 정상대조군(Vehicle)을 제외한 나머지 실험군에 1 mM H2O2를 처리한 후 48시간 동안 배양하였다. 그 후 세포 분쇄액이 희석된 완충액을 샘플로 하여 샘플 50 ㎕에 MES 버퍼, 보조인자 믹스(cofactor mixture), 효소 믹스, DTNB를 혼합한 반응 버퍼(reaction buffer) 150 ㎕를 넣고 암실에서 가볍게 섞어준 후, 30분 이내로, 405~414 nm 범위에서 흡광도를 측정하였다.HaCaT cells were aliquoted to 5×10 6 cells/ml on a cell culture plate (SPL, 100 cm 2 ). After culturing for 48 hours, the composition of the present invention was treated, and thereafter, 1 mM H 2 O 2 was treated in the remaining experimental groups except for the normal control group (Vehicle), and then cultured for 48 hours. Then, using the buffer solution diluted with the cell crushing solution as a sample, 150 μl of a reaction buffer mixed with MES buffer, cofactor mixture, enzyme mix, and DTNB was added to 50 μl of the sample and mixed lightly in the dark. Then, within 30 minutes, the absorbance was measured in the range of 405 to 414 nm.
그 결과, 도 5d에 나타낸 바와 같이, 본 발명의 조성물 처리 농도 증가에 따라 농도 의존적으로 GSH 발현이 증가하는 경향을 보이며, 100 μg/ml 농도에서 H2O2 단독처리군 대비 GSH 발현이 23% 개선됨을 확인하였다. 동일농도(100 μg/ml)의 홍삼 단독처리군과 비교할 때에는 GSH 발현이 23% 증가함을 확인하였다. 상기 결과를 통해, 항산화 방어기작으로 발현이 증가하는 GSH가 본 발명의 조성물에 의하여 증가하며, 그 효과는 홍삼 단독처리군보다 뛰어남을 확인하였다.As a result, as shown in FIG. 5d , the GSH expression tends to increase in a concentration-dependent manner as the concentration of the composition of the present invention increases, and at a concentration of 100 μg/ml, the GSH expression is 23% compared to the H 2 O 2 alone treatment group. Improvement was confirmed. It was confirmed that the expression of GSH increased by 23% when compared with the group treated with red ginseng alone at the same concentration (100 μg/ml). Through the above results, it was confirmed that GSH, which is increased in expression as an antioxidant defense mechanism, is increased by the composition of the present invention, and the effect is superior to that of the red ginseng alone treatment group.
상기 결과를 통해, 본 발명의 조성물은 항산화 효과가 우수하며, 오메가 7 및 홍삼의 시너지 효과에 의해 단독물질 처리 시보다 그 효과가 뛰어남을 확인하였다.Through the above results, it was confirmed that the composition of the present invention has excellent antioxidant effect, and the effect is superior to that of treatment with a single substance due to the synergistic effect of
실험예 5. 본 발명 조성물의 항염증 효과 확인Experimental Example 5. Confirmation of the anti-inflammatory effect of the composition of the present invention
5-1. IL-1β, TNF-α 저해 활성5-1. IL-1β, TNF-α inhibitory activity
ELISA 키트를 이용하여 HaCaT 세포에서 분비되는 항염증 인자, 즉 IL-1β, TNF-α의 농도 변화를 통한 본 발명 조성물의 우수한 항염증 효과를 확인하였다. 상기 HaCaT 세포주는 ATCC에서 구매하여 실험을 진행하였고, 세포는 DMEM 배지, 10% 소태아혈청, 50 U/ml P/S(페니실린/스트렙토마이신)에서 37℃, 습도 95%, 5%의 CO2, CO2 인큐베이터(Sanyo, Japan) 조건 하에서 배양한 세포주를 실험에 사용하였다. The excellent anti-inflammatory effect of the composition of the present invention was confirmed by changing the concentration of the anti-inflammatory factors secreted from HaCaT cells, that is, IL-1β and TNF-α using the ELISA kit. The HaCaT cell line was purchased from ATCC and the experiment was performed, and the cells were prepared in DMEM medium, 10% fetal bovine serum, 50 U/ml P/S (penicillin/streptomycin) at 37°C, humidity 95%, 5% CO 2 , CO 2 Cell lines cultured under conditions of an incubator (Sanyo, Japan) were used for the experiment.
구체적으로, 우선 상기 HaCaT 세포를 2×105 세포/웰의 농도로 분주하여 배양하였다. 24시간 동안 배양한 후, 본 발명의 조성물을 100 nl/ml 농도로 처리하였고, 이후 정상대조군(Vehicle)을 제외한 나머지 실험군에 1 mM H2O2를 처리한 후, 24시간 동안 배양하였다. HaCaT 세포 배양액을 샘플로 하여 IL-1β 및 TNF-α 생성량을 측정 하였다. Specifically, the HaCaT cells were first aliquoted at a concentration of 2×10 5 cells/well and cultured. After culturing for 24 hours, the composition of the present invention was treated at a concentration of 100 nl/ml, and then the remaining experimental groups except for the normal control group (Vehicle) were treated with 1 mM H 2 O 2 , and then cultured for 24 hours. Using the HaCaT cell culture medium as a sample, IL-1β and TNF-α production levels were measured.
IL-1β 및 TNF-α 생성량은 인간 IL-1β ELISA 키트(Abcam, Cambridge, MA)와 인간 TNF-α ELISA 키트 (Abcam, Cambridge, MA)를 이용하여 수행하였다. 구체적으로, 96-웰 플레이트에 차례대로 HaCaT 세포 배양액 100 ㎕를 넣고 실온에서 2시간 30분 동안 배양하였다. 이 후 바이오틴으로 표지된 IL-1β 및 TNF-α 검출 항체(각 항체는 키트에서 제공하는 항체를 사용)를 100 ㎕ 첨가한 후 1시간 동안 배양하였다. 이후, 상기 배양물을 세척한 후, HRP-스트렙타비딘 용액 100 ㎕를 첨가하였으며, 추가적으로 실온에서 30분간 배양하였다. 이 후 50 ㎕의 정지 용액(stop solution)을 첨가한 후, 450 nm의 흡광도에서 실험 결과를 측정하였다. TNF-α에 대한 결과는 도 6a에 나타내었고, IL-1β에 대한 결과는 도 6b에 나타내었다. IL-1β and TNF-α production levels were performed using a human IL-1β ELISA kit (Abcam, Cambridge, MA) and a human TNF-α ELISA kit (Abcam, Cambridge, MA). Specifically, 100 μl of the HaCaT cell culture solution was sequentially added to a 96-well plate and incubated at room temperature for 2 hours and 30 minutes. Thereafter, 100 μl of biotin-labeled IL-1β and TNF-α detection antibodies (each antibody using an antibody provided in the kit) was added and incubated for 1 hour. Then, after washing the culture, 100 μl of HRP-streptavidin solution was added, and incubated for 30 minutes at room temperature. Thereafter, 50 μl of a stop solution was added, and the experimental results were measured at absorbance at 450 nm. Results for TNF-α are shown in FIG. 6a, and results for IL-1β are shown in FIG. 6b.
도 6a 및 도 6b에 나타낸 바와 같이, H2O2에 의한 활성산소 증가로 인하여 염증반응이 일어나며, 본 발명의 조성물 처리 농도 증가에 따라 농도 의존적으로 TNF-α, IL-1β의 농도가 감소하고, 100 μg/ml 농도에서 H2O2 단독처리군 대비 TNF-α, IL-1β 저해 활성이 각각 46%, 33% 증가함을 확인하였다. 동일농도(100 μg/ml)의 홍삼 단독처리군과 비교할 때에는 TNF-α, IL-1β 저해 활성이 각각 25%, 12% 증가함을 확인하였다. 상기 결과를 통해, 본 발명의 조성물이 우수한 항염증 효과를 가짐을 확인하였다.As shown in FIGS. 6a and 6b, an inflammatory reaction occurs due to an increase in active oxygen by H 2 O 2 , and the concentration of TNF-α, IL-1β decreases in a concentration-dependent manner according to the increase in the treatment concentration of the composition of the present invention, and , it was confirmed that the TNF-α, IL-1β inhibitory activity was increased by 46% and 33%, respectively, compared to the H 2 O 2 alone treatment group at a concentration of 100 μg/ml. It was confirmed that the TNF-α, IL-1β inhibitory activity increased by 25% and 12%, respectively, when compared with the group treated with red ginseng at the same concentration (100 μg/ml). Through the above results, it was confirmed that the composition of the present invention has an excellent anti-inflammatory effect.
5-2. COX-2, PGE2 저해 활성5-2. COX-2, PGE2 inhibitory activity
웨스턴 블로팅을 통해 염증인자인 COX-2, PGE2 단백질 발현량을 측정하여 본 발명 조성물의 항염증 활성을 확인하고자 하였다. HaCaT 세포를 5×106 cell/ml의 농도로 10% FBS를 첨가한 DMEM 배양액에 분주한 후, 세포를 12시간 동안 안정화시켰다. 세포에 시료를 처리한 후 1시간 동안 유지하고, H2O2 (1mM) 처리 또는 자외선조사를 진행한 후, 10% FBS가 함유된 DMEM 배지와 함께 24시간 배양하였다. 배양한 세포에 RIPA 버퍼를 처리하고, 4℃, 15,000 RPM, 15분의 조건으로 원심분리하였다. 분리한 단백질 샘플은 아크릴아마이드 젤(acrylamide gel) 8-12%에서 전기영동 후 PVDF(Polyvinylidene Difluoride) 막에 옮기고, 실온에서 블로킹 버퍼(PBS-T에 5% 탈지분유)를 사용하여 블로킹한 뒤 COX-2, PGE2를 1:1,000 비율로 희석하여 4℃에서 24시간 동안 인큐베이션하였다. 반응이 끝나면, 세척 후 실온에서 2차 항체와 1시간동안 반응시킨 후, SWDED 기질(Supersignal West Dura extended duration substrate; Thermo, Rockford, CA, USA)을 사용한 화학발광 방법(Chemi Imager analyzer system; Alpha Innotech, San Leandro, CA, USA)을 통하여 결과를 확인하였다.It was attempted to confirm the anti-inflammatory activity of the composition of the present invention by measuring the expression levels of COX-2 and PGE2 proteins, which are inflammatory factors, through Western blotting. HaCaT cells were aliquoted in DMEM culture medium supplemented with 10% FBS at a concentration of 5×10 6 cell/ml, and then the cells were stabilized for 12 hours. Cells were maintained for 1 hour after processing the sample, and after treatment with H 2 O 2 (1 mM) or UV irradiation, the cells were incubated with DMEM medium containing 10% FBS for 24 hours. The cultured cells were treated with RIPA buffer, and centrifuged at 4° C., 15,000 RPM, and 15 minutes. The separated protein sample is electrophoresed in 8-12% acrylamide gel, then transferred to a PVDF (Polyvinylidene Difluoride) membrane, blocked using a blocking buffer (5% skim milk in PBS-T) at room temperature, and then COX -2, PGE2 was diluted at a ratio of 1:1,000 and incubated at 4°C for 24 hours. After the reaction, after washing and reacting with the secondary antibody at room temperature for 1 hour, a chemiluminescence method (Chemi Imager analyzer system; Alpha Innotech) using SWDED substrate (Supersignal West Dura extended duration substrate; Thermo, Rockford, CA, USA) , San Leandro, CA, USA) to confirm the results.
그 결과, 도 7a에 나타낸 바와 같이, 본 발명의 조성물을 처리한 경우, 처리 농도가 증가할수록 COX-2의 발현량이 농도 의존적으로 감소하는 경향을 보이며, 50~100 μg/ml 농도 범위에서 H2O2 단독처리군 대비 유의하게 발현이 저해됨을 확인하였다. 100 μg/ml 농도에서는 H2O2 단독처리군 대비 COX-2 발현이 55% 감소함을 확인하였다.As a result, as shown in FIG. 7a, when the composition of the present invention was treated, the expression level of COX-2 showed a tendency to decrease in a concentration-dependent manner as the treatment concentration increased, and H 2 in the concentration range of 50-100 μg/ml O 2 It was confirmed that the expression was significantly inhibited compared to the single treatment group. At a concentration of 100 μg/ml, it was confirmed that the expression of COX-2 was reduced by 55% compared to the H 2 O 2 alone treatment group.
또한, 도 7b에 나타낸 바와 같이, 본 발명의 조성물을 처리한 경우 처리 농도가 증가할수록 PGE2의 발현량이 농도 의존적으로 감소하는 경향을 보이며, 100 μg/ml 농도에서 H2O2 단독처리군 대비 유의하게 발현이 저해됨을 확인하였다. 100 μg/ml 처리 농도에서는 H2O2 단독처리군 대비 PGE2 발현이 79% 감소함을 확인하였다.In addition, as shown in FIG. 7b , when the composition of the present invention was treated, the expression level of PGE2 showed a tendency to decrease in a concentration-dependent manner as the treatment concentration increased, compared to the H 2 O 2 alone treatment group at 100 μg/ml concentration. It was confirmed that the expression was inhibited. At 100 μg/ml treatment concentration, it was confirmed that PGE2 expression was reduced by 79% compared to H 2 O 2 alone treatment group.
상기 결과를 통해, 본 발명의 조성물은 산화적 스트레스에 의한 항염증 인자(COX-2, PGE2)의 단백질 발현을 현저히 억제하는 효과가 있음을 확인하였다.Through the above results, it was confirmed that the composition of the present invention has an effect of significantly inhibiting the protein expression of anti-inflammatory factors (COX-2, PGE2) caused by oxidative stress.
5-3. MMP-1, MMP-3 저해 활성 및 PCOLA1, Elastin 증가 활성5-3. MMP-1, MMP-3 inhibitory activity and PCOLA1, Elastin-increasing activity
웨스턴 블로팅을 통해 HaCaT 세포에서 분비되는 단백질인 MMP-1, MMP-3, PCOLA1(프로콜라겐 유형 1), Elastin(엘라스틴)의 발현량을 측정하여 본 발명 조성물의 우수한 피부 탄력 및 주름 개선 효과를 확인하고자 하였다. 본 실험은 COX-2 및 PGE2에 대한 항체를 사용하는 것 대신, MMP-1, 프로콜라겐 유형 1(PCOLA1) 및 엘라스틴에 대한 항체(cell signalling)를 사용하여 실험을 수행한 것 외에는 상기 실험예 5-2와 동일한 방식으로 수행하였다. 그 결과를 도 8에 나타내었다.By measuring the expression levels of MMP-1, MMP-3, PCOLA1 (procollagen type 1), and Elastin (elastin), which are proteins secreted from HaCaT cells, through Western blotting, the excellent skin elasticity and wrinkle improvement effects of the composition of the present invention were evaluated. wanted to confirm. In this experiment, instead of using antibodies against COX-2 and PGE2, Experimental Example 5 except that the experiment was performed using antibodies to MMP-1, procollagen type 1 (PCOLA1) and elastin (cell signaling) -2 was performed in the same manner. The results are shown in FIG. 8 .
도 8a 및 8b에 나타낸 바와 같이, 본 발명의 조성물을 처리한 경우 처리 농도가 증가할수록 MMP-1, MMP-3의 발현량이 농도 의존적으로 감소하는 경향을 보이며, 100 μg/ml 처리 농도에서 H2O2 단독처리군 대비 각각 74%, 48% 감소됨을 확인하였다. 홍삼 단일처리군과 비교할 때에는 MMP-1, MMP-3 발현 저해 효과가 각각 19%, 16% 높은 것으로 확인하였다.As shown in FIGS. 8a and 8b , when the composition of the present invention was treated, the expression levels of MMP-1 and MMP-3 tended to decrease in a concentration-dependent manner as the treatment concentration increased, and H 2 at 100 μg/ml treatment concentration It was confirmed that 74% and 48% reduction compared to the O 2 alone treatment group, respectively. When compared with the red ginseng single treatment group, it was confirmed that the inhibitory effects of MMP-1 and MMP-3 were 19% and 16% higher, respectively.
또한, 도 8c 및 8d에 나타낸 바와 같이, 본 발명의 조성물을 처리한 경우 처리 농도가 증가할수록 PCOLA1, Elastin의 발현량이 농도 의존적으로 증가하는 경향을 보이며, 100 μg/ml 처리 농도에서 H2O2 단독처리군 대비 각각 44%, 59% 증가함을 확인하였다. 홍삼 단일처리군과 비교할 때에는 PCOLA-1, Elastin 발현 증가 효과가 각각 17%, 14% 높은 것으로 확인하였다.In addition, as shown in FIGS. 8c and 8d , when the composition of the present invention was treated, the expression levels of PCOLA1 and Elastin tended to increase in a concentration-dependent manner as the treatment concentration increased, and H 2 O 2 at 100 μg/ml treatment concentration It was confirmed that the increase was 44% and 59%, respectively, compared to the single treatment group. When compared with the red ginseng single treatment group, it was confirmed that the effect of increasing the expression of PCOLA-1 and Elastin was 17% and 14% higher, respectively.
상기 결과를 통해, H2O2에 의한 활성산소 증가로 인하여 염증반응이 발생한 세포에 본 발명의 조성물을 처리하는 경우 상기 화학적 자극에 의해 발생한 MMP-1, MMP-3의 발현 수준이 현저히 감소하고, 피부에서 콜라겐을 합성하고 피부 탄성 유지 역할을 하는 PCOLA1, Elastin의 수준이 현저하게 증가함을 확인하였다. 이는 본 발명의 조성물이 피부 탄력 및 주름 개선 효과를 가지고 있어, 종합적으로 피부 상태를 개선시키는 효과를 가짐을 나타낸다. Through the above results, when the composition of the present invention is treated to the cells in which the inflammatory reaction has occurred due to the increase in active oxygen by H 2 O 2 The expression levels of MMP-1 and MMP-3 generated by the chemical stimulation are significantly reduced and , It was confirmed that the levels of PCOLA1 and Elastin, which synthesize collagen in the skin and maintain skin elasticity, were remarkably increased. This indicates that the composition of the present invention has an effect of improving skin elasticity and wrinkles, and has an effect of improving the skin condition overall.
상기 실험 결과는 Student's t-test와 ANOVA를 동시에 수행하여 나타낸 것이며, p값이 0.05 미만일 경우, 통계학적으로 유의성이 있다고 판정하였다.The experimental results are shown by simultaneously performing Student's t-test and ANOVA, and when the p value is less than 0.05, it was determined to be statistically significant.
상기 결과를 통해, 본 발명의 조성물은 높은 안정성을 기반으로 하여, 오메가-7과 홍삼을 혼합함에 따른 시너지 효과로서 피부 보습도, 피부보호장벽 개선, 산화적 스트레스에 의한 세포손상 저해 효과를 가져 미세먼지와 같은 외부 자극에 의한 피부의 손상에 대하여 종합적인 피부 상태 개선 효과를 가짐을 확인하였다. 뿐만 아니라, 본 발명의 조성물은 상기 효과들과 더불어 항산화, 항염증 효과, 피부 탄력 및 주름 개선 효과를 가지고 있어, 우수한 피부 노화 억제 효과를 가진다. 이에 따라 본 발명의 조성물은 단순한 식품부터 건강기능식품 및 화장품 등으로 다양하게 활용할 수 있다.Based on the above results, the composition of the present invention has a synergistic effect by mixing omega-7 and red ginseng, based on high stability, to improve skin moisture, improve skin barrier, and inhibit cell damage caused by oxidative stress. It was confirmed that it has a comprehensive skin condition improvement effect on skin damage caused by external stimuli such as dust. In addition, the composition of the present invention has an antioxidant, anti-inflammatory effect, skin elasticity and wrinkle improvement effect in addition to the above effects, and has an excellent skin aging inhibitory effect. Accordingly, the composition of the present invention can be used in a variety of ways, from simple foods to health functional foods and cosmetics.
이하 본 발명의 조성물을 포함하는 식품 조성물 및 화장료 조성물의 제제예를 설명한다.Hereinafter, formulation examples of food compositions and cosmetic compositions containing the composition of the present invention will be described.
제제예 1. 식품 제제의 제조Formulation Example 1. Preparation of food formulations
1. 건강식품의 제조1. Manufacture of health food
본 발명의 조성물 100 mg100 mg of the composition of the present invention
비타민 혼합물 적량appropriate amount of vitamin mixture
비타민 A 아세테이트 70 g 70 g vitamin A acetate
비타민 E 1.0 mgVitamin E 1.0 mg
비타민 B1 0.13 mgVitamin B1 0.13 mg
비타민 B2 0.15 mgVitamin B2 0.15 mg
비타민 B6 0.5 mg0.5 mg of vitamin B6
비타민 B12 0.2 g 0.2 g of vitamin B12
비타민 C 10 mg
비오틴 10 g 10 g of biotin
니코틴산아미드 1.7 mg1.7 mg of nicotinic acid amide
엽산 50 g 50 g folic acid
판토텐산 칼슘 0.5 mgCalcium pantothenate 0.5 mg
무기질 혼합물 적량Mineral mixture appropriate amount
황산제1철 1.75 mgferrous sulfate 1.75 mg
산화아연 0.82 mgZinc Oxide 0.82 mg
탄산마그네슘 25.3 mgMagnesium carbonate 25.3 mg
제1인산칼륨 15 mgpotassium phosphate monobasic 15 mg
제2인산칼슘 55 mgDicalcium Phosphate 55 mg
구연산칼륨 90 mgPotassium citrate 90 mg
탄산칼슘 100 mg100 mg of calcium carbonate
염화마그네슘 24.8 mgMagnesium chloride 24.8 mg
상기의 비타민 및 미네랄 혼합물의 조성비는 비교적 건강식품에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상의 건강식품 제조방법에 따라 상기의 성분을 혼합한 다음, 과립을 제조하고, 통상의 방법에 따라 건강식품 조성물 제조에 사용할 수 있다.The composition ratio of the above vitamin and mineral mixture is a composition that is relatively suitable for health food in a preferred embodiment, but the mixing ratio may be arbitrarily modified. , to prepare granules, and can be used for preparing health food compositions according to a conventional method.
2. 건강음료의 제조2. Manufacture of health drinks
본 발명의 조성물 100 mg100 mg of the composition of the present invention
비타민 C 15 g15 g vitamin C
비타민 E(분말) 100 g100 g vitamin E (powder)
젖산철 19.75 g19.75 g of iron lactate
산화아연 3.5 g3.5 g zinc oxide
니코틴산아미드 3.5 g3.5 g of nicotinic acid amide
비타민 A 0.2 g0.2 g vitamin A
비타민 B1 0.25 g0.25 g of vitamin B1
비타민 B2 0.3 g0.3 g of vitamin B2
물 정량water metering
통상의 건강음료 제조방법에 따라 상기의 성분을 혼합한 다음, 약 1 시간 동안 85 에서 교반 가열한 후, 만들어진 용액을 여과하여 멸균된 2 L 용기에 취득하여 밀봉 멸균한 뒤 냉장 보관한 다음 본 발명의 건강음료 조성물 제조에 사용한다.After mixing the above ingredients according to a conventional health drink manufacturing method, after stirring and heating at 85 for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized, and then refrigerated. used in the manufacture of health beverage compositions.
상기 조성비는 비교적 기호음료에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만 수요계층이나, 수요국가, 사용용도 등 지역적, 민족적 기호도에 따라서 그 배합비를 임의로 변형 실시하여도 무방하다.Although the composition ratio is prepared by mixing ingredients suitable for relatively favorite beverages in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and national preferences such as demanding class, demanding country, and use.
제제예 2. 화장료 제제의 제조Formulation Example 2. Preparation of a cosmetic formulation
1. 유연화장수 제조1. Manufacture of softened lotion
본 발명의 조성물 0.1중량%0.1% by weight of the composition of the present invention
1,3-부틸렌글리콜 5.2중량%1,3-butylene glycol 5.2 wt%
올레일알코올 1.5중량%1.5 wt% of oleyl alcohol
에탄올 3.2중량%3.2% by weight of ethanol
폴리솔베이트 20 3.2중량%
벤조페논-9 2.0중량%benzophenone-9 2.0 wt%
카르복실비닐폴리머 1.0중량%1.0 wt% of carboxyl vinyl polymer
글리세린 3.5중량%Glycerin 3.5% by weight
미량의 향, 미량의 방부제 및 잔량의 정제수를 혼합하여 통상의 방법으로 유연화장수를 제조하였다.A small amount of fragrance, a small amount of preservative, and a residual amount of purified water were mixed to prepare a softening lotion in a conventional manner.
2. 밀크로션 제조2. Milk lotion production
본 발명의 조성물 0.1중량%0.1% by weight of the composition of the present invention
글리세린 5.1중량%5.1% by weight of glycerin
프로필렌글리콜 4.2중량%4.2 wt% of propylene glycol
토코페릴아세테이트 3.0중량%3.0 wt% tocopheryl acetate
유동파라핀 4.6중량%Liquid paraffin 4.6 wt%
트리에탄올아민 1.0중량%1.0% by weight of triethanolamine
스쿠알란 3.1중량%3.1% by weight of squalane
마카다미아너트오일 2.5중량%2.5% by weight of macadamia nut oil
폴리솔베이트 60 1.6중량%
솔비탄세스퀴롤레이트 1.6중량%1.6 wt% of sorbitan sesquirolate
프로필파라벤 0.6중량%Propylparaben 0.6 wt%
카르복실비닐폴리머 1.5중량%1.5 wt% of carboxyl vinyl polymer
미량의 향, 미량의 방부제, 잔량의 정제수를 혼합하여 통상의 방법으로 밀크로션을 제조하였다.A small amount of fragrance, a small amount of preservative, and the remaining amount of purified water were mixed to prepare a milk lotion by a conventional method.
3. 영양크림 제조3. Manufacture of nourishing cream
본 발명의 조성물 0.5중량%0.5% by weight of the composition of the present invention
글리세린 4.0중량%Glycerin 4.0% by weight
바셀린 3.5중량%Vaseline 3.5% by weight
트리에탄올아민 2.1중량%2.1 wt% of triethanolamine
유동파라핀 5.3중량%Liquid paraffin 5.3 wt%
스쿠알란 3.0중량%3.0 wt% squalane
밀납 2.6중량%Beeswax 2.6 wt%
토코페릴아세테이트 5.4중량%Tocopheryl acetate 5.4 wt%
폴리솔베이트 60 3.2중량%
카르복실비닐폴리머 1.0중량%1.0% by weight of carboxyl vinyl polymer
솔비탄세스퀴올레이트 3.1중량%3.1 wt% of sorbitan sesquioleate
미량의 향, 미량의 방부제 및 잔량의 정제수를 혼합하여 통상의 방법으로 영양크림을 제조하였다.A small amount of flavor, a small amount of preservative, and the remaining amount of purified water were mixed to prepare a nourishing cream by a conventional method.
4. 마사지크림 제조4. Manufacture of massage cream
본 발명의 조성물 0.5중량%0.5% by weight of the composition of the present invention
글리세린 4.0중량%Glycerin 4.0% by weight
바셀린 3.5중량%Vaseline 3.5% by weight
트리에탄올아민 0.5중량%0.5 wt% of triethanolamine
유동파라핀 24.0중량%Liquid paraffin 24.0 wt%
스쿠알란 3.0중량%3.0 wt% squalane
밀납 2.1중량%Beeswax 2.1 wt%
토코페릴아세테이트 0.1중량%Tocopheryl acetate 0.1% by weight
폴리솔베이트 60 2.4중량%
카르복실비닐폴리머 1.0중량%1.0% by weight of carboxyl vinyl polymer
솔비탄세스퀴올레이트 2.3중량%Sorbitan sesquioleate 2.3 wt%
미량의 향, 미량의 방부제 및 잔량의 정제수를 혼합하여 통상의 방법으로 마사지크림을 제조하였다.A small amount of fragrance, a small amount of preservative, and the remaining amount of purified water were mixed to prepare a massage cream in a conventional manner.
Claims (11)
A health functional food composition for improving skin condition comprising omega-7 derived from refined fish oil and red ginseng mixed in a weight ratio of 2:8 to 5:5 (w/w).
상기 오메가-7은 오메가-7 함유 정제어유의 형태로 상기 조성물에 포함되는 것을 특징으로 하는, 조성물.
According to claim 1,
The omega-7 is characterized in that included in the composition in the form of omega-7-containing refined fish oil, the composition.
상기 정제어유는 정제된 폴락 오일(refined pollock oil)인 것을 특징으로 하는, 조성물.
6. The method of claim 5,
The refined fish oil is a refined pollock oil (refined pollock oil), characterized in that the composition.
상기 피부상태 개선은 피부 자극에 의한 피부 손상 억제 또는 피부 표면막 개선에 의한 것인, 조성물.
According to claim 1,
The improvement of the skin condition is due to inhibition of skin damage caused by skin irritation or improvement of the skin surface film, the composition.
상기 피부 자극은 물리적 자극 또는 화학적 자극인 것을 특징으로 하는, 조성물.
8. The method of claim 7,
The composition, characterized in that the skin irritation is a physical stimulus or a chemical stimulus.
A food additive for improving skin condition comprising omega-7 derived from refined fish oil and red ginseng mixed in a weight ratio of 2:8 to 5:5 (w/w).
A cosmetic composition for improving skin condition comprising omega-7 derived from refined fish oil and red ginseng mixed in a weight ratio of 2:8 to 5:5 (w/w).
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KR101066676B1 (en) | 2005-05-20 | 2011-09-21 | (주)아모레퍼시픽 | A oral composition for improving beauty of skin |
KR101338140B1 (en) * | 2011-02-25 | 2013-12-06 | 사단법인 진안군친환경홍삼한방산업클러스터사업단 | A pharmaceutical comprising the extract of Red Panax ginseng and Fish oil for treating or preventing skin aging |
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Non-Patent Citations (2)
Title |
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학위논문(송인봉, 'OMEGA_7의 피부 항노화 효과에 대한 연구', 전북대학교, 2018년 02월 22일) |
한방안이비인후피부과학회지 제27권 제3호 (통권 57호), 106_114쪽. |
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