KR102683009B1 - Manufacturing method fo cosmetics composition - Google Patents
Manufacturing method fo cosmetics composition Download PDFInfo
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- KR102683009B1 KR102683009B1 KR1020230060621A KR20230060621A KR102683009B1 KR 102683009 B1 KR102683009 B1 KR 102683009B1 KR 1020230060621 A KR1020230060621 A KR 1020230060621A KR 20230060621 A KR20230060621 A KR 20230060621A KR 102683009 B1 KR102683009 B1 KR 102683009B1
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- peony flower
- extract
- peony
- fermented
- lactobacillus
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- 239000000203 mixture Substances 0.000 title claims abstract description 16
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- WZNJWVWKTVETCG-UHFFFAOYSA-N kojic acid Natural products OC(=O)C(N)CN1C=CC(=O)C(O)=C1 WZNJWVWKTVETCG-UHFFFAOYSA-N 0.000 description 2
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- YKRGDOXKVOZESV-WRJNSLSBSA-N Paeoniflorin Chemical compound C([C@]12[C@H]3O[C@]4(O)C[C@](O3)([C@]1(C[C@@H]42)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)C)OC(=O)C1=CC=CC=C1 YKRGDOXKVOZESV-WRJNSLSBSA-N 0.000 description 1
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- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Biotechnology (AREA)
- Engineering & Computer Science (AREA)
- Dermatology (AREA)
- Botany (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Cosmetics (AREA)
- Medicines Containing Plant Substances (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
본 발명은 피부 항산화, 미백, 항염증 및 주름 등 피부 상태 개선 효과가 우수한 화장료 조성물의 제조방법에 관한 것으로, 본 발명의 화장료 조성물은 모란꽃을 락토바실러스 균주로 발효한 모란꽃 발효 추출물을 함유한다.The present invention relates to a method for producing a cosmetic composition that has excellent skin condition improvement effects such as skin antioxidant, whitening, anti-inflammatory, and wrinkle. The cosmetic composition of the present invention contains a fermented peony flower extract obtained by fermenting peony flowers with Lactobacillus strains.
Description
본 발명은 화장료 조성물의 제조 방법에 관한 것으로, 특히 락토바실러스(Lactobacillus) 균주를 이용한 모란꽃 발효 추출물을 포함하는 피부 개선용 화장료 조성물의 제조 방법에 관한 것이다.The present invention relates to a method for producing a cosmetic composition, and in particular, to a method for producing a cosmetic composition for improving skin containing a fermented peony flower extract using Lactobacillus strains.
현대 사회에서 신체의 건강과 외모가 경쟁력 중 하나라는 인식과 함께 화장품에 대한 관심은 남녀 구분 없이 높아지고 있으며, 생활 수준의 질적 향상과 더불어 단순히 외모를 가꾸는 의미를 넘어서 정신적, 신체적 건강을 함께 고려하여 화장품과 미용용품을 선택하고 있다 (Global Market Report(2015), Natural Cosmetics Market Trends in Major Countries, KOTRA, 15-032). In modern society, with the recognition that physical health and appearance are one of competitiveness, interest in cosmetics is increasing regardless of gender, and with the qualitative improvement in the standard of living, cosmetics are being developed by taking mental and physical health into consideration beyond simply taking care of one's appearance. and beauty products (Global Market Report (2015), Natural Cosmetics Market Trends in Major Countries, KOTRA, 15-032).
피부가 자외선을 받으면 멜라닌 색소가 생성되어 일차적으로 피부에서 발생하는 활성산소와 프리 라디칼(free radical)을 제거해 주고, 자외선을 흡수 차단시켜 피부를 보호하기도 하지만, 멜라닌 자체가 활성산소를 발생시키기도 하며 피부노화 진행을 가속화 시킨다.When the skin receives ultraviolet rays, melanin pigment is produced, which primarily removes active oxygen and free radicals generated in the skin and protects the skin by absorbing and blocking ultraviolet rays, but melanin itself also generates active oxygen and damages the skin. Accelerates the aging process.
이러한 프리 라디칼을 제거하기 위해 지금까지 BHA 및 BHT 등의 합성 항산화제가 개발되어 식품, 화장품 등에 산화방지제로 많이 사용되었으나, 안전성에 논란이 있어 허용 대상 식품에서는 사용량이 엄격히 규제되어 보다 안전하고 강한 항산화제의 개발이 요구되는 실정이다(Choe SY. and Yang KH(1982), Toxicological studies of antioxidant, butylated hydroxytoluene(BHT) and butylated hydroxyanisole(BHA), Korean J. Hood Sci. Technol, 12:283-288). To remove these free radicals, synthetic antioxidants such as BHA and BHT have been developed and widely used as antioxidants in foods and cosmetics, etc. However, due to controversy over their safety, the amount of use in permitted foods is strictly regulated, resulting in safer and stronger antioxidants. The development of is required (Choe SY. and Yang KH (1982), Toxicological studies of antioxidant, butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA), Korean J. Hood Sci. Technol, 12:283-288).
특히 피부에 작용하는 화장품의 경우 다양한 부작용이 발생할 수 있기 때문에 천연 기능성 소재에 대한 중요성이 더욱 높아지고 있으며, 그 중에서 미백 효과 및 항산화 작용을 통한 노화 방지에 대한 관련 연구가 다양하게 이루어지고 있다(Natural Cosmetics Industry Trend Report(2013), The Foundation of AG. Tech, Commercialization and Transfer). In particular, the importance of natural functional materials is increasing because various side effects can occur in the case of cosmetics that act on the skin. Among them, various studies are being conducted on whitening effects and anti-aging through antioxidant effects (Natural Cosmetics Industry Trend Report (2013), The Foundation of AG.
현재 알려진 미백 원료 중 하이드로퀴논(hydroquinone)은 멜라닌 형성에서 티로시나제 (tyrosinase) 활성을 저해하여 멜라닌 생합성 초기 단계에서 티로신의 산화 과정을 방해함으로써 색소 형성 및 침착을 개선하지만 정상 피부 세포의 섬유 모세포에 대해 독성을 보인다고 알려져 있다(Yim SH, Cho KS, Choi JH, Lee JH, Kim MS, Lee BHN(2016), Effect of various pear cultivars at different fruit development stages on antioxidant and whitening activities. Korean J Food Sci Technol, 48:59-65). Among currently known whitening raw materials, hydroquinone improves pigment formation and deposition by inhibiting tyrosinase activity in melanin formation and interfering with the oxidation process of tyrosine in the early stage of melanin biosynthesis, but is toxic to fibroblasts of normal skin cells. (Yim SH, Cho KS, Choi JH, Lee JH, Kim MS, Lee BHN (2016), Effect of various pear cultivars at different fruit development stages on antioxidant and whitening activities. Korean J Food Sci Technol, 48: 59-65).
다른 미백 원료인 비타민 C(vitamin C)와 코직산(kojic acid) 또한 강력한 미백 효과가 있는 것으로 알려져 있으나, 비타민 C는 유산소 조건하에서 매우 불안정하기 때문에 제한적으로 사용되며, 코직산의 경우 알레르기 유발 가능성이 있어 화장품 원료로 사용이 어려운 실정이다. Other whitening ingredients, vitamin C and kojic acid, are also known to have a strong whitening effect, but vitamin C is very unstable under aerobic conditions, so its use is limited, and kojic acid has the potential to cause allergies, so it is used in cosmetics. It is difficult to use it as a raw material.
이와 같은 문제점들로 인하여 천연의 물질로부터 미백 활성을 갖는 물질의 개발에 노력하고 있다.Due to these problems, efforts are being made to develop substances with whitening activity from natural substances.
본 발명은 락토바실러스(Lactobacillus) 균주를 이용한 모란꽃 발효 추출물을 포함하여 피부 항산화, 미백, 항염증 및 주름개선 효과가 더 우수한 피부 개선용 화장료 조성물의 제조 방법을 제공하는 것을 목적으로 한다.The purpose of the present invention is to provide a method for producing a cosmetic composition for skin improvement that includes a fermented peony flower extract using a Lactobacillus strain and has better skin antioxidant, whitening, anti-inflammatory and wrinkle improvement effects.
상기 목적을 달성하기 위한 본 발명의 실시예에 따른 화장료 조성물은, 모란꽃을 락토바실러스 균주로 발효한 모란꽃 발효 추출물을 함유함에 그 특징이 있다. The cosmetic composition according to an embodiment of the present invention for achieving the above object is characterized by containing a fermented peony flower extract obtained by fermenting peony flowers with a Lactobacillus strain.
상기 락토바실러스 균주는, 락토바실러스 아시도필루스, 락토바실러스 플란타룸, 락토바실러스 펜토서스, 락토바실러스 브레비스, 락토바실러스 카제이 군으로부터 선택되는 1종 이상인 것을 특징으로 한다.The Lactobacillus strain is characterized as being at least one selected from the group of Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus pentosus, Lactobacillus brevis, and Lactobacillus casei.
한편, 상기와 같은 목적을 달성하기 위한 본 발명의 실시예에 따른 화장료 조성물의 제조 방법은, 모란꽃 분말에 에탄올을 가하여 모란꽃 추출물을 형성하는 단계; 상기 모란꽃 추출물을 락토바실러스 균주를 이용하여 발효하여 모란꽃 발표물을 형성하는 단계; 그리고, 상기 모란꽃 발효물을 여과하여 모란꽃 발효 추출물을 형성하는 단계를 포함함에 그 특징이 있다.Meanwhile, a method for producing a cosmetic composition according to an embodiment of the present invention to achieve the above object includes the steps of adding ethanol to peony flower powder to form a peony flower extract; Forming a peony flower extract by fermenting the peony flower extract using a Lactobacillus strain; And, it is characterized by comprising the step of filtering the peony flower fermentation product to form a peony flower fermentation extract.
상기 모란꽃 추출물을 형성하는 단계는, 상기 모란꽃 분말에 에탄올을 가하고, 70℃ 내지 90℃에서 2시간 내지 4시간 동안 환류 추출하는 단계와, 상기 환류 추출한 후 감압 농축하는 단계를 포함하는 단계를 포함함을 특징으로 한다.The step of forming the peony flower extract includes adding ethanol to the peony flower powder, refluxing and extracting at 70°C to 90°C for 2 to 4 hours, and concentrating under reduced pressure after the reflux extraction. It is characterized by .
상기 모란꽃 발표물을 형성하는 단계는, 상기 모란꽃 추출물에 락토바실러스 균주를 최종 농도 0.5×106 CFU/㎖ 내지 1.5×106 CFU/㎖ 가 되도록 접종한 후 pH 5.0 ~ 6.0 및 34℃ 내지 40℃에서 6일 내지 8일 동안 발효하는 단계와, 상기 발효물을 90℃ ~ 100℃ 온도에서 20분 내지 40분 조건에서 실활하여 냉침하는 단계를 포함하는 것을 특징으로 한다.In the step of forming the peony flower extract, the peony flower extract is inoculated with Lactobacillus strain to a final concentration of 0.5 × 10 6 CFU/ml to 1.5 × 10 6 CFU/ml, and then incubated at pH 5.0 to 6.0 and 34°C to 40°C. It is characterized in that it includes the step of fermenting for 6 to 8 days, and the step of deactivating and cold immersing the fermented product at a temperature of 90 ℃ to 100 ℃ for 20 to 40 minutes.
상기 락토바실러스 균주는, 락토바실러스 아시도필루스, 락토바실러스 플란타룸, 락토바실러스 펜토서스, 락토바실러스 브레비스, 락토바실러스 카제이 군으로부터 선택되는 1종 이상인 것을 특징으로 한다.The Lactobacillus strain is characterized as being at least one selected from the group of Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus pentosus, Lactobacillus brevis, and Lactobacillus casei.
본 발명의 실시예에 따른 화장료 조성물 및 그의 제조 방법에 있어서는 다음과 같은 효과가 있다.The cosmetic composition and its manufacturing method according to an embodiment of the present invention have the following effects.
모란꽃 분말에 에탄올을 가하고 일정 온도에서 일정 시간 동안 환류 추출한 후 감압 농축하여 추출물을 형성한 후, 상기 모란꽃 추출물을 균주를 이용하여 발효 추출하므로, 피부 항산화, 미백, 항염증 및 주름개선 효과가 더 우수한 발효추출물을 얻을 수 있다. Ethanol is added to peony flower powder, refluxed and extracted at a certain temperature for a certain time, and then concentrated under reduced pressure to form an extract. The peony flower extract is then fermented and extracted using a strain, resulting in superior skin antioxidant, whitening, anti-inflammatory and wrinkle improvement effects. Fermentation extract can be obtained.
또한, 상기 모란꽃 발효 추출물을 유효 성분으로 화장료 베이스에 적용하였을 때 피부 상태 개선 효과가 우수한 화장료 조성물을 제공할 수 있다.In addition, when the fermented peony flower extract is applied as an active ingredient to a cosmetic base, a cosmetic composition with excellent skin condition improvement effect can be provided.
도 1은 본 발명의 [실시예 1] 내지 [실시예 5]로부터 수득된 모란꽃 발효 추출물의 DPPH (2,2-Diphenyl-1-1-picrylhydrazyl) 소거 활성 결과를 [비교예 1]과 비교하여 나타낸 그래프이다.
도 2는 본 발명의 [실시예 1] 내지 [실시예 5]로부터 수득된 모란꽃 발효 추출물의 COX-2 (Cycloxygenase-2) 억제 활성 결과를 [비교예 1]과 비교하여 나타낸 그래프이다.
도 3은 본 발명의 [실시예 1] 내지 [실시예 5]로부터 수득된 모란꽃 발효 추출물의 세포 생존율(Cell viability) 결과를 [비교예 1]과 비교하여 나타낸 그래프이다.
도 4는 본 발명의 [실시예 1] 내지 [실시예 5]로부터 수득된 모란꽃 발효 추출물의 티로시나제(Tyronsinase) 결과를 [비교예 1]과 비교하여 나타낸 그래프이다.Figure 1 shows the DPPH (2,2-Diphenyl-1-1-picrylhydrazyl) scavenging activity results of peony flower fermentation extracts obtained from [Example 1] to [Example 5] of the present invention compared with [Comparative Example 1]. This is the graph shown.
Figure 2 is a graph showing the COX-2 (Cycloxygenase-2) inhibitory activity results of peony flower fermented extracts obtained from [Example 1] to [Example 5] of the present invention compared with [Comparative Example 1].
Figure 3 is a graph showing the cell viability results of the fermented peony flower extracts obtained from [Example 1] to [Example 5] of the present invention compared with [Comparative Example 1].
Figure 4 is a graph showing the tyrosinase results of the fermented peony flower extract obtained from [Example 1] to [Example 5] of the present invention compared with [Comparative Example 1].
이하, 상기와 같은 특징을 갖는 본 발명의 바람직한 실시예에 따른 피부 개선용 화장료 조성물 및 그의 제조 방법을 첨부 도면을 참조하여 보다 상세히 설명하면 다음과 같다. Hereinafter, the cosmetic composition for skin improvement and its manufacturing method according to a preferred embodiment of the present invention having the above-mentioned characteristics will be described in more detail with reference to the accompanying drawings.
먼저, 모란(Paeonia suffruticosa)은 작약과 작약속의 낙엽 활엽 떨기나무다. 목단, 부귀화라고도 한다. 오래전부터 화단이나 정원에 관상용으로 심었다. 꽃이 풍성하고 아름다워 과거에는 '꽃 중의 왕'이란 뜻의 '화중지왕(花中之王)' 혹은 '나라에서 가장 빼어난 향'이란 뜻의 '국색천향(國色天香)' 등으로 불렸다. First, peony (Paeonia suffruticosa) is a deciduous broadleaf tree of the peony genus. It is also called mokdan or wealth. It has long been planted as an ornamental plant in flower beds or gardens. Because the flowers are abundant and beautiful, in the past it was called 'Hwajungjiwang (花中之王)', meaning 'king of flowers', or 'Guksaekcheonhyang (國色天香)', meaning 'the most outstanding fragrance in the country'.
한국에서도 오래전부터 꽃을 감상하거나 뿌리를 약으로 쓰기 위해 심었다. 뿌리 껍질을 말린 목단피는 한방에서 소염, 진통, 정혈, 고혈압 등에 쓰고 있다. 모란은 상징성이나 관상학적뿐만 아니라, 생리학적 가치도 뛰어나다. 모란의 뿌리 껍질을 의미하는 목단피(牡丹皮)는 동의 보감에 다양한 병증을 치료하는 주요한 한약재로 기록되어 있다. In Korea, it has been planted for a long time to enjoy the flowers or to use the roots as medicine. Mokdan bark, dried root bark, is used in oriental medicine for anti-inflammatory, pain-relieving, blood purification, and high blood pressure. Peonies have outstanding symbolic and physiognomic value as well as physiological value. Mokdanpi (牡丹皮), which refers to the root bark of peony, is recorded in Dongbogam as an important herbal medicine for treating various diseases.
모란의 주요 성분으로는 페오놀 배당체인 페오노시드, 페오놀리드, 페오니플로린, 옥시페오니플로린, 탄닌 등으로 알려져 있다.The main components of peony are known to be pheonol glycosides, pheonoside, pheonolide, peoniflorin, oxypheoniflorin, and tannin.
한편, 발효는 가장 오래된 역사를 가진 생물학의 기술로 산업 전반에 걸친 기술 개발이 현재에도 이루어지고 있다. 특히 식품에서의 기술 개발이 주류를 이루고 있으며, 의약품의 경우 미생물에서 생산되는 약물 흡수 촉진 성분인 지질당체, 인지질, 지질단백질복합체 등의 미생물 유래 성분이 약물의 생체흡수를 개선하여 생체 약리효과를 더욱 증진시킬 수 있다고 보고되어 있다. Meanwhile, fermentation is a biological technology with the longest history, and technological development across industries is still taking place. In particular, technology development in food is mainstream, and in the case of pharmaceuticals, microbial-derived components such as lipid sugars, phospholipids, and lipoprotein complexes, which are drug absorption promoting components produced by microorganisms, improve the bioabsorption of drugs, further enhancing biopharmacological effects. It has been reported that it can be improved.
발효 물질은 천연 물질 고분자를 생전환시켜 저분자 물질의 구조로 분해되어 체내 흡수율을 높일 뿐 아니라 생체 이용률이 증가해 효과가 빠른 장점을 가지고 있다.Fermented substances have the advantage of being effective quickly by bioconverting natural polymers and decomposing them into low-molecular structure structures, thereby increasing absorption into the body and increasing bioavailability.
이에 다양한 천연 소재를 발효시키게 되면 저분자 물질의 활성화로 인해 다양한 기능성이 증진되어 그 효과와 이용가치가 더욱 높아 진다.Accordingly, when various natural materials are fermented, various functionalities are enhanced due to the activation of low-molecular-weight substances, further increasing the effectiveness and utility value.
상기 모란을 이용한 발효 기술을 이용하여 본 발명의 실시예에 따른 모란꽃 발효 추출물을 포함하는 피부 개선용 화장료 조성물 및 그의 제조 방법을 보다 구체적으로 설명하면 다음과 같다.A cosmetic composition for improving skin containing a fermented peony flower extract according to an embodiment of the present invention using the fermentation technology using the peony and its manufacturing method will be described in more detail as follows.
[비교예 1][Comparative Example 1]
모란꽃(Paeonia suffruticosa) 분말 20g 내지 30g (예를 들면, 25g)에 50% 내지 90% (예를 들면, 70%) 에탄올 220ml 내지 280ml (예를 들면, 250ml)를 가하고, 70℃ 내지 90℃ (예를 들면, 80℃)에서 2시간 내지 4시간 (예를 들면, 3시간) 동안 환류 추출한 후 감압 농축하여 추출물을 제조한다.220ml to 280ml (eg, 250ml) of 50% to 90% (eg, 70%) ethanol was added to 20g to 30g (eg, 25g) of peony flower (Paeonia suffruticosa) powder, and incubated at 70°C to 90°C ( For example, an extract is prepared by performing reflux extraction at 80° C. for 2 to 4 hours (for example, 3 hours) and then concentrating under reduced pressure.
제조된 [비교예 1]의 모란꽃 추출물에 락토바실러스 펜토서스(Lactobacillus pentosus) 균주를 최종 농도 0.5×106 CFU/㎖ 내지 1.5×106 CFU/㎖ (예를 들면, 1×106 CFU/㎖)가 되도록 접종한 후 pH 5.0 ~ 6.0 및 34℃ 내지 40℃ (예를 들면 37℃)에서 6일 내지 8일 (예를 들면 7일) 동안 발효한다. Lactobacillus pentosus strain was added to the peony flower extract prepared in [Comparative Example 1] at a final concentration of 0.5×10 6 CFU/mL to 1.5×10 6 CFU/mL (for example, 1×10 6 CFU/mL). ) and then fermented for 6 to 8 days (e.g. 7 days) at pH 5.0 to 6.0 and 34°C to 40°C (e.g. 37°C).
상기 발효물을 90℃ ~ 100℃, 20분 내지 40분 (예를 들면 30분) 조건에서 실활하여 냉침한 후 와트만(Whatman) No. 2 여과지를 이용하여 모란꽃 발효 추출물을 제조한다.The fermented product was deactivated and cold soaked at 90°C to 100°C for 20 to 40 minutes (e.g., 30 minutes) and then soaked in Whatman No. 2 Prepare peony flower fermentation extract using filter paper.
이를 락토바실러스 펜토서스 균주를 이용한 발효 추출법이라 한다.This is called a fermentation extraction method using Lactobacillus pentosus strains.
제조된 [비교예 1] 모란꽃 추출물에 락토바실러스 브레비스(Lactobacillus brevis) 균주를 최종 농도 0.5×106 CFU/㎖ 내지 1.5×106 CFU/㎖ (예를 들면, 1×106 CFU/㎖)가 되도록 접종한 후 pH 5.0 ~ 6.0 및 34℃ 내지 40℃ (예를 들면 37℃)에서 6일 내지 8일 (예를 들면 7일) 동안 발효한다. Prepared [Comparative Example 1] Lactobacillus brevis strain was added to the peony flower extract at a final concentration of 0.5×10 6 CFU/mL to 1.5×10 6 CFU/mL (for example, 1×10 6 CFU/mL). After inoculation, fermentation is carried out for 6 to 8 days (eg, 7 days) at pH 5.0 to 6.0 and 34°C to 40°C (eg, 37°C).
상기 발효물을 90℃ ~ 100℃, 20분 내지 40분 (예를 들면 30분) 조건에서 실활하여 냉침한 후 와트만(Whatman) No. 2 여과지를 이용하여 모란꽃 발효 추출물을 제조한다.The fermented product was deactivated and cold soaked at 90°C to 100°C for 20 to 40 minutes (e.g., 30 minutes) and then soaked in Whatman No. 2 Prepare peony flower fermentation extract using filter paper.
이를 락토바실러스 브레비스 균주를 이용한 발효 추출법이라 한다.This is called a fermentation extraction method using Lactobacillus brevis strains.
제조된 [비교예 1] 모란꽃 추출물에 락토바실러스 플라타룸(Lactobacillus plantarum) 균주를 최종 농도 0.5×106 CFU/㎖ 내지 1.5×106 CFU/㎖ (예를 들면, 1×106 CFU/㎖)가 되도록 접종한 후 pH 5.0 ~ 6.0 및 34℃ 내지 40℃ (예를 들면 37℃)에서 6일 내지 8일 (예를 들면 7일) 동안 발효한다. Prepared [Comparative Example 1] Lactobacillus plantarum strain was added to the peony flower extract at a final concentration of 0.5×10 6 CFU/ml to 1.5×10 6 CFU/ml (for example, 1×10 6 CFU/ml). After inoculation, it is fermented for 6 to 8 days (eg, 7 days) at pH 5.0 to 6.0 and 34°C to 40°C (eg, 37°C).
상기 발효물을 90℃ ~ 100℃, 20분 내지 40분 (예를 들면 30분) 조건에서 실활하여 냉침한 후 와트만(Whatman) No. 2 여과지를 이용하여 모란꽃 발효 추출물을 제조한다.The fermented product was deactivated and cold soaked at 90°C to 100°C for 20 to 40 minutes (e.g., 30 minutes), then soaked in Whatman No. 2 Prepare peony flower fermentation extract using filter paper.
이를 락토바실러스 플란타룸 균주를 이용한 발효 추출법이라 한다.This is called a fermentation extraction method using Lactobacillus plantarum strains.
제조된 [비교예 1] 모란꽃 추출물에 락토바실러스 카제이(Lactobacillus casei) 균주를 최종 농도 0.5×106 CFU/㎖ 내지 1.5×106 CFU/㎖ (예를 들면, 1×106 CFU/㎖)가 되도록 접종한 후 pH 5.0 ~ 6.0 및 34℃ 내지 40℃ (예를 들면 37℃)에서 6일 내지 8일 (예를 들면 7일) 동안 발효한다. Prepared [Comparative Example 1] Lactobacillus casei strain was added to the peony flower extract at a final concentration of 0.5×10 6 CFU/ml to 1.5×10 6 CFU/ml (for example, 1×10 6 CFU/ml). After inoculation, it is fermented for 6 to 8 days (eg, 7 days) at pH 5.0 to 6.0 and 34°C to 40°C (eg, 37°C).
상기 발효물을 90℃ ~ 100℃, 20분 내지 40분 (예를 들면 30분) 조건에서 실활하여 냉침한 후 와트만(Whatman) No. 2 여과지를 이용하여 모란꽃 발효 추출물을 제조한다.The fermented product was deactivated and cold soaked at 90°C to 100°C for 20 to 40 minutes (e.g., 30 minutes) and then soaked in Whatman No. 2 Prepare peony flower fermentation extract using filter paper.
이를 락토바실러스 카제이 균주를 이용한 발효 추출법이라 한다.This is called a fermentation extraction method using Lactobacillus casei strain.
제조된 [비교예1] 모란꽃 추출물에 락토바실러스 아시도필루스(Lactobacillus acidophilus) 균주를 최종 농도 0.5×106 CFU/㎖ 내지 1.5×106 CFU/㎖ (예를 들면, 1×106 CFU/㎖)가 되도록 접종한 후 pH 5.0 ~ 6.0 및 34℃ 내지 40℃ (예를 들면 37℃)에서 6일 내지 8일 (예를 들면 7일) 동안 발효한다. Prepared [Comparative Example 1] Lactobacillus acidophilus strain was added to the peony flower extract at a final concentration of 0.5 × 10 6 CFU / ㎖ to 1.5 × 10 6 CFU / ㎖ (for example, 1 × 10 6 CFU / ㎖) and then fermented for 6 to 8 days (eg, 7 days) at pH 5.0 to 6.0 and 34°C to 40°C (eg, 37°C).
상기 발효물을 90℃ ~ 100℃, 20분 내지 40분 (예를 들면 30분) 조건에서 실활하여 냉침한 후 와트만(Whatman) No. 2 여과지를 이용하여 모란꽃 발효 추출물을 제조한다.The fermented product was deactivated and cold soaked at 90°C to 100°C for 20 to 40 minutes (e.g., 30 minutes) and then soaked in Whatman No. 2 Prepare peony flower fermentation extract using filter paper.
이를 락토바실러스 아시도필루스 균주를 이용한 발효 추출법이라 한다.This is called a fermentation extraction method using Lactobacillus acidophilus strains.
도 1은 본 발명의 [실시예 1] 내지 [실시예 5]로부터 수득된 모란꽃 발효 추출물의 DPPH (2,2-Diphenyl-1-1-picrylhydrazyl) 소거 활성 결과를 [비교예 1]과 비교하여 나타낸 그래프이고, 도 2는 본 발명의 [실시예 1] 내지 [실시예 5]로부터 수득된 모란꽃 발효 추출물의 COX-2 (Cycloxygenase-2) 억제 활성 결과를 [비교예 1]과 비교하여 나타낸 그래프이다.Figure 1 shows the DPPH (2,2-Diphenyl-1-1-picrylhydrazyl) scavenging activity results of peony flower fermentation extracts obtained from [Example 1] to [Example 5] of the present invention compared with [Comparative Example 1]. It is a graph shown, and Figure 2 is a graph showing the COX-2 (Cycloxygenase-2) inhibitory activity results of the peony flower fermentation extract obtained from [Example 1] to [Example 5] of the present invention compared with [Comparative Example 1]. am.
도 3은 본 발명의 [실시예 1] 내지 [실시예 5]로부터 수득된 모란꽃 발효 추출물의 세포 생존율(Cell viability) 결과를 [비교예 1]과 비교하여 나타낸 그래프이고, 도 4는 본 발명의 [실시예 1] 내지 [실시예 5]로부터 수득된 모란꽃 발효 추출물의 티로시나제(Tyronsinase) 결과를 [비교예 1]과 비교하여 나타낸 그래프이다.Figure 3 is a graph showing the cell viability results of the peony flower fermentation extract obtained from [Example 1] to [Example 5] of the present invention compared with [Comparative Example 1], and Figure 4 is a graph showing the cell viability results of the peony flower fermentation extract obtained from [Example 1] to [Example 5] of the present invention. This is a graph showing the tyrosinase results of the fermented peony flower extracts obtained from [Example 1] to [Example 5] compared with [Comparative Example 1].
[시험예 1][Test Example 1]
DPPH에 의한 자유라디칼 소거법에 따른 환원 활성Reduction activity according to free radical scavenging method by DPPH
상기 [실시예 1] 내지 [실시예 5]의 방법으로 얻어진 모란꽃 발효 추출물을 [비교예 1]의 방법으로 얻어진 모란꽃 에탄올 추출물과 비교하여 DPPH 라디칼(radical) 활성 효과를 측정하였다.The effect of DPPH radical activity was measured by comparing the fermented peony flower extract obtained by the method of [Example 1] to [Example 5] with the ethanol extract of peony flower obtained by the method of [Comparative Example 1].
1,1-디페닐-2-피크릴-히드라진 (DPPH, 1,1-diphenyl-2-picryl hydrazyl, Sigma, USA)은 화학적으로 안정화된 자유 라디칼을 가지고 있는 수용성 물질로 520nm 부근에서 최대 흡광도를 가지며, 항산화 활성이 있는 물질과 만나면 전자를 내어주면서 라디칼 (DPPH)이 소멸되고 색깔이 변한다. 화학적으로 안정성 있는 DPPH은 여러 종의 항산화 성분이 내재된 추출물, 음료와 오일, 순수 페놀화합물 등의 항산화 효과를 분석할 수 있다. 이 실험은 DPPH로 먼저 산화를 개시한 후 시료에 의해서 라디칼에 대한 소거되는 활성을 알아보기 위한 실험이다. DPPH를 메탄올에 용해시킨 0.2mM의 DPPH 용액 1mL에 각 농도의 [비교예 1]의 에탄올 추출물과 [실시예 1] 내지 [실시예 5]의 발효 추출물을 각각 25, 50, 100, 250, 500, 1000 ㎍/㎖ 농도 별로 첨가하였다. 실온에서 10분간 반응시킨 후 ELISA reader (Plus384, Molecular Device, USA)로 525nm에서 흡광도를 측정하였다. 대조군은 시료 액과 DPPH 용액 대신 메탄올을 넣어 보정 값을 얻는다.1,1-diphenyl-2-picryl-hydrazyl (DPPH, 1,1-diphenyl-2-picryl hydrazyl, Sigma, USA) is a water-soluble substance with chemically stabilized free radicals and has a maximum absorbance around 520 nm. When it comes into contact with a substance with antioxidant activity, it donates electrons, causing the radical (DPPH) to disappear and its color to change. DPPH, which is chemically stable, can analyze the antioxidant effect of extracts containing various antioxidant components, beverages and oils, and pure phenolic compounds. This experiment is to first initiate oxidation with DPPH and then examine the radical scavenging activity of the sample. In 1 mL of a 0.2mM DPPH solution in which DPPH was dissolved in methanol, the ethanol extract of [Comparative Example 1] and the fermentation extract of [Example 1] to [Example 5] of each concentration were added at 25, 50, 100, 250, and 500, respectively. , 1000 ㎍/㎖ was added at each concentration. After reacting at room temperature for 10 minutes, absorbance was measured at 525 nm with an ELISA reader (Plus384, Molecular Device, USA). In the control group, correction values are obtained by adding methanol instead of the sample solution and DPPH solution.
자유 라디칼 소거율은 아래의 식에 따라 계산하였다.The free radical scavenging rate was calculated according to the formula below.
소거율 (%) = (대조군의 흡광도-시험군의 흡광도) / 대조군의 흡광도 × 100Clearance rate (%) = (absorbance of control group - absorbance of test group) / absorbance of control group × 100
대조군은 비타민 C(L-ascorbic acid, Sigma, USA)를 사용하였다. 활성은 50%로 기준하였으며 이와 비슷하거나 높은 값을 가지는 경우 활성이 높은 것으로 간주했다.Vitamin C (L-ascorbic acid, Sigma, USA) was used as the control group. Activity was standard at 50%, and if the value was similar or higher, the activity was considered high.
그 결과, 상기 [표 1] 및 도 1에서 나타낸 바와 같이, [비교예 1]과 [실시예 1] 내지 [실시예 5]의 농도 별로 처리하였을 때 락토바실러스로 발효 추출한 모란꽃이 에탄올 추출보다 약 60~ 70% 활성이 더 우수한 DPPH 소거능을 나타내어 높은 항산화 활성을 나타내었다.As a result, as shown in [Table 1] and FIG. 1, when treated at different concentrations in [Comparative Example 1] and [Example 1] to [Example 5], peony flowers fermented and extracted with Lactobacillus were less effective than those extracted with ethanol. 60~70% activity showed superior DPPH scavenging ability, showing high antioxidant activity.
[시험예 2][Test Example 2]
항염증 - COX-2인자 확인Anti-inflammatory - COX-2 factor confirmation
상기 [실시예 1] 내지 [실시예 5]의 방법으로 얻어진 모란꽃 발효 추출물과 [비교예 1]의 방법으로 얻어진 모란꽃 에탄올 추출물을 각각 25, 50, 100, 250, 500, 1000 ㎍/㎖ 농도 별로 항염증(COX-2) 활성 효과를 측정하였다. The peony flower fermentation extract obtained by the method of [Example 1] to [Example 5] and the peony flower ethanol extract obtained by the method of [Comparative Example 1] were administered at concentrations of 25, 50, 100, 250, 500, and 1000 μg/ml, respectively. The anti-inflammatory (COX-2) activity effect was measured.
Raw 264.7 세포 5 × 105 개를 60 mm culture dish에 분주하고 18시간 동안 배양하였다. 배양한 Raw 264.7 세포에 새 DMEM (10% FBS) 배지로 교체한 후, 2시간 동안 샘플을 전처리 하였다. 2시간 뒤 1 ㎍/㎖의 LPS를 처리하여 4시간 동안 자극 시켰다. 4시간 후 세포를 harverst 하여 NucleoZOL lysis buffer(NM, 740404)를 처리하고, NM에서 제공하는 protocol을 이용하여 RNA isolation을 진행 하였으며, 분리된 RNA를 microplate reader를 이용하여 정량한 뒤 HiSenScript™ RH(-) RT PreMix Kit(intron, 25087)을 이용하여 cDNA를 합성하였다. PCR master mix(Bioneer, K-2016)을 이용하여 유전자를 증폭한 후 증폭 산물을 정량 분석하였다. 5 × 10 5 Raw 264.7 cells were dispensed into a 60 mm culture dish and cultured for 18 hours. The cultured Raw 264.7 cells were replaced with new DMEM (10% FBS) medium, and the samples were pretreated for 2 hours. After 2 hours, the cells were treated with 1 μg/ml LPS and stimulated for 4 hours. After 4 hours, the cells were harvested and treated with NucleoZOL lysis buffer (NM, 740404). RNA isolation was performed using the protocol provided by NM. The isolated RNA was quantified using a microplate reader and then incubated with HiSenScript™ RH (- ) cDNA was synthesized using the RT PreMix Kit (intron, 25087). After amplifying the gene using PCR master mix (Bioneer, K-2016), the amplification product was quantitatively analyzed.
대조군은 EGCG (Epigallocatechin gallate, Sigma, USA)를 사용하였다. 활성은 50%로 기준하였으며 이와 비슷하거나 높은 값을 가지는 경우 활성이 높은 것으로 간주했다.EGCG (Epigallocatechin gallate, Sigma, USA) was used as the control group. Activity was standard at 50%, and if the value was similar or higher, the activity was considered high.
그 결과, 상기 [표 2] 및 도 2에서 나타낸 바와 같이, [비교예 1]과 [실시예 1] 내지 [실시예 5]의 농도 별로 처리하였을 때 락토바실러스로 발효 추출한 모란꽃이 에탄올 추출보다 약 50~ 60% 더 우수한 COX-2 억제율을 나타내었으며, 유의성 있는 항염 효과를 확인할 수 있었다.As a result, as shown in [Table 2] and FIG. 2, when treated at different concentrations in [Comparative Example 1] and [Example 1] to [Example 5], peony flowers fermented and extracted with Lactobacillus were less effective than those extracted with ethanol. It showed a 50-60% better COX-2 inhibition rate, and a significant anti-inflammatory effect was confirmed.
[시험예 3][Test Example 3]
MTT assay를 이용한 세포독성 측정Cytotoxicity measurement using MTT assay
상기 [실시예 1] 내지 [실시예 5]의 방법으로 얻어진 모란꽃 발효 추출물과 [비교예 1]의 방법으로 얻어진 모란꽃 에탄올 추출물을 각각 25, 50, 100, 250, 500, 1000 ㎍/㎖ 농도별로 세포독성을 MTT assay로 측정하였다.The peony flower fermentation extract obtained by the method of [Example 1] to [Example 5] and the peony flower ethanol extract obtained by the method of [Comparative Example 1] were administered at concentrations of 25, 50, 100, 250, 500, and 1000 μg/ml, respectively. Cytotoxicity was measured by MTT assay.
세포를 3-[4,5-dimethyltjiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT, Amresco)으로 염색시켜 세포의 생존율을 측정한다. 96well plate에 HaCaT 세포를 6×103 cells/well 로 분주하고 24시간 배양한다. 각 시료를 25, 50, 100, 250, 500, 1000 ㎍/㎖로 처리하고 72시간 배양한다. MTT(5 ㎎/㎖)를 4시간 동안 처리한 후, ELISA reader (Plus 384, Molecular Device, USA)로 570nm에서 흡광도를 측정하였다. 세포 생존율은 80% 이상이면 독성이 없고 80% 이하이면 독성이 있다고 간주했다.Cell viability is measured by staining cells with 3-[4,5-dimethyltjiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT, Amresco). HaCaT cells were distributed at 6×10 3 cells/well in a 96-well plate and cultured for 24 hours. Each sample was treated with 25, 50, 100, 250, 500, and 1000 ㎍/㎖ and incubated for 72 hours. After treatment with MTT (5 mg/ml) for 4 hours, absorbance was measured at 570 nm with an ELISA reader (Plus 384, Molecular Device, USA). Cell viability was considered non-toxic if it was over 80% and toxic if it was less than 80%.
그 결과, 상기 [표 3] 및 도 3에서 나타낸 바와 같이, [비교예 1]과 [실시예 1] 내지 [실시예 5]의 사용 농도가 최고 농도인 1000 ㎍/㎖ 에서도 세포독성이 없는 것으로 나타난 것으로 보아, 독성이 없는 안전한 원료로 사용이 가능함을 확인하였다.As a result, as shown in [Table 3] and Figure 3, there was no cytotoxicity even at the highest concentration of 1000 μg/ml in [Comparative Example 1] and [Example 1] to [Example 5]. Based on what was shown, it was confirmed that it can be used as a safe, non-toxic raw material.
[시험예 4][Test Example 4]
미백 효능 검증 시험: 티로시나아제(Tyrosinase) 활성Whitening efficacy verification test: Tyrosinase activity
상기 [실시예 1] 내지 [실시예 5]의 방법으로 얻어진 모란꽃 발효 추출물과 [비교예 1]의 방법으로 얻어진 모란꽃 에탄올 추출물을 각각 25, 50, 100, 250, 500, 1000 ㎍/㎖ 농도별로 티로시나아제(Tyrosinase) 활성 효과를 측정하였다.The peony flower fermentation extract obtained by the method of [Example 1] to [Example 5] and the peony flower ethanol extract obtained by the method of [Comparative Example 1] were administered at concentrations of 25, 50, 100, 250, 500, and 1000 μg/ml, respectively. Tyrosinase activity effect was measured.
멜라닌은 tyrosinase가 tyrosine을 산화하여 생성이 시작된다. 각 sample의 tyrosinase와 경쟁적으로 억제를 하여 미백 효능을 검증하는 방법으로 Yagi의 방법을 변형하여 시행하였다. Melanin production begins when tyrosinase oxidizes tyrosine. Yagi's method was modified to verify whitening efficacy by competitively inhibiting tyrosinase of each sample.
실험 방법은 3,4- Dihydroxyphenylalanine(L-DOPA, Sigma, USA)를 10 mM의 농도를 가진0.1 M Phosphate buffer(pH 6.8) 0.5 ㎖와 sample을 농도별로 0.1 M Phosphate buffer(pH 6.8) 혼합액 0.5 ㎖을 첨가하였다. 여기에 mushroom tyrosinase (110 U/㎖, Sigma co. USA) 0.2 mL를 첨가하여 37℃ incubator에서 5분간 반응시킨 후. 475 nm 파장에서 흡광도를 측정하였다. sample을 첨가하지 않은 동일한 혼합물을 대조구로 사용하였다. Tyrosinase 효소 활성 저해율은 다음 계산식에 의하여 산출하였다.The experimental method is 0.5 ml of 0.1 M Phosphate buffer (pH 6.8) containing 3,4- Dihydroxyphenylalanine (L-DOPA, Sigma, USA) at a concentration of 10 mM, and 0.5 ml of 0.1 M Phosphate buffer (pH 6.8) mixture for each concentration. was added. Add 0.2 mL of mushroom tyrosinase (110 U/mL, Sigma co. USA) and react in an incubator at 37°C for 5 minutes. Absorbance was measured at a wavelength of 475 nm. The same mixture without added sample was used as a control. Tyrosinase enzyme activity inhibition rate was calculated using the following formula.
% 저해율 = (A-B)/A×100% inhibition rate = (A-B)/A×100
A: 시료를 첨가하지 않았을 때의 흡광도A: Absorbance when no sample is added
B: 시료를 첨가하였을 때의 흡광도B: Absorbance when sample is added
대조군은 알부틴(Arbutin, Sigma, USA)를 사용하였다. 활성은 50%로 기준하였으며 이와 비슷하거나 높은 값을 가지는 경우 활성이 높은 것으로 간주했다.Arbutin (Arbutin, Sigma, USA) was used as the control group. Activity was standard at 50%, and if the value was similar or higher, the activity was considered high.
그 결과, 상기 [표 4] 및 도 4에서 나타낸 바와 같이 [비교예 1]과 [실시예 1] 내지 [실시예 5]의 농도 별로 처리하였을 때 비교예 및 실시예 모두 농도 의존적으로 티로시나아제(Tyrosinase) 활성을 억제하는 효과를 나타내었으나, 락토바실러스로 발효 추출한 모란꽃이 에탄올 추출보다 약 26 ~ 33% 더 우수한 티로시나아제 활성 억제율을 나타내어 더 높은 미백 효과를 확인하였다.As a result, as shown in [Table 4] and Figure 4, when [Comparative Example 1] and [Example 1] to [Example 5] were treated at different concentrations, both Comparative Examples and Examples showed tyrosinase in a concentration-dependent manner. (Tyrosinase) activity was inhibited, but peony flowers fermented and extracted with Lactobacillus showed a tyrosinase activity inhibition rate that was about 26 to 33% better than ethanol extracted, confirming a higher whitening effect.
이상에서 설명한 바와 같이, 모란꽃 분말에 에탄올을 가하고 일정 온도에서 일정 시간 동안 환류 추출한 후 감압 농축하여 추출물을 형성한 후, 상기 모란꽃 추출물을 균주를 이용하여 발효 추출한 모란꽃 발효 추출물을 유효성분으로 포함하는 피부 상태 개선용 화장료를 제조할 수 있다.As described above, ethanol was added to peony flower powder, refluxed and extracted at a certain temperature for a certain time, concentrated under reduced pressure to form an extract, and then the peony flower extract was fermented and extracted using a strain. Skin containing the peony flower fermented extract as an active ingredient. Cosmetics for improving condition can be manufactured.
따라서, 이와 같이 제조된 피부 상태 개선용 화장료는 피부 항산화, 미백, 항염증 및 주름 개선 효과가 우수하고, 피부 상태 개선 우수한 효과를 얻을 수 있다.Therefore, the cosmetic for improving skin condition prepared in this way has excellent skin antioxidant, whitening, anti-inflammatory and wrinkle improvement effects, and can achieve excellent skin condition improvement effects.
이상의 설명은 본 발명을 예시적으로 설명한 것에 불과하며, 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자에 의해 본 발명의 기술적 사상에서 벗어나지 않는 범위에서 다양한 변형이 가능할 것이다. 따라서 본 발명의 명세서에 개시된 실시 예들은 본 발명을 한정하는 것이 아니다. 본 발명의 범위는 아래의 특허청구범위에 의해 해석되어야 하며, 그와 균등한 범위 내에 있는 모든 기술도 본 발명의 범위에 포함되는 것으로 해석해야 할 것이다.The above description is merely an exemplary description of the present invention, and various modifications may be made by those skilled in the art without departing from the technical spirit of the present invention. Accordingly, the embodiments disclosed in the specification of the present invention do not limit the present invention. The scope of the present invention should be interpreted in accordance with the scope of the patent claims below, and all technologies within the equivalent scope thereof should be interpreted as being included in the scope of the present invention.
삭제delete
Claims (6)
상기 모란꽃 추출물에 락토바실러스 펜토서스 균주를 최종 농도 0.5×106 CFU/㎖ 내지 1.5×106 CFU/㎖ 가 되도록 접종한 후, pH 5.0 ~ 6.0 및 34℃ 내지 40℃에서 6일 내지 8일 동안 발효하고, 상기 발효물을 90℃ ~ 100℃ 온도에서 20분 내지 40분 조건에서 실활하여 냉침하여 모란꽃 발효물을 형성하는 단계; 그리고,
상기 모란꽃 발효물을 여과하여 모란꽃 발효 추출물을 형성하는 단계를 포함하는 화장료 조성물의 제조 방법.Adding ethanol to peony flower powder, performing reflux extraction at 70°C to 90°C for 2 to 4 hours, and then concentrating under reduced pressure to form a peony flower extract;
The peony flower extract was inoculated with Lactobacillus pentosus strain to a final concentration of 0.5×10 6 CFU/ml to 1.5×10 6 CFU/ml, and then incubated at pH 5.0 to 6.0 and 34°C to 40°C for 6 to 8 days. Fermenting, deactivating and cold soaking the fermented product at a temperature of 90°C to 100°C for 20 to 40 minutes to form a peony flower fermentation product; and,
A method for producing a cosmetic composition comprising filtering the peony flower fermentation product to form a peony flower fermentation extract.
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| KR102351384B1 (en) | 2021-10-21 | 2022-01-14 | (주)에버코스 | Cosmetic composition containing fermented extract using paeonia suffruticosa, camphor tree leaf, vitamin tree leaf, centella asiatica and method for manufacturing the same |
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| KR102351384B1 (en) | 2021-10-21 | 2022-01-14 | (주)에버코스 | Cosmetic composition containing fermented extract using paeonia suffruticosa, camphor tree leaf, vitamin tree leaf, centella asiatica and method for manufacturing the same |
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