TWI788019B - Use of chamaemelum nobile extract for manufacturing skincare composition - Google Patents
Use of chamaemelum nobile extract for manufacturing skincare composition Download PDFInfo
- Publication number
- TWI788019B TWI788019B TW110134713A TW110134713A TWI788019B TW I788019 B TWI788019 B TW I788019B TW 110134713 A TW110134713 A TW 110134713A TW 110134713 A TW110134713 A TW 110134713A TW I788019 B TWI788019 B TW I788019B
- Authority
- TW
- Taiwan
- Prior art keywords
- skin
- extract
- chrysanthemum
- composition
- chamaemelum nobile
- Prior art date
Links
Images
Landscapes
- Cosmetics (AREA)
Abstract
Description
本案係關於一種果香菊( Chamaemelum nobile)萃取物的用途,特別是關於果香菊萃取物用於製備調理肌膚的組合物之用途。 This case is about the use of an extract of Chamaemelum nobile , especially the use of the extract of Chamaemelum nobile for preparing a skin conditioning composition.
肌膚會隨年齡增長而正常老化,在現代生活中,人們承受著各式各樣的壓力,作息不正常、熬夜、睡眠不足、日夜顛倒、飲食失衡等生理和心理上的壓力是造成肌膚泛紅及膚色不均的主要原因,人們只能通過彩妝掩蓋肌膚瑕疵,然而,彩妝的使用加劇了肌膚的受損程度,使肌膚彈力下降、保水度不足、粗燥、無光澤、毛孔粗大、皺紋、肌膚產生斑點及/或整體較為暗沈、蠟黃等老化態樣。此外,這些壓力造成肌膚發生代謝紊亂,使肌膚無法及時分解和排出代謝廢物,大量堆積在細胞中。速肌膚老化的速度,使整體外觀看起來無精打采。The skin will age normally with age. In modern life, people are under various pressures, such as abnormal work and rest, staying up late, lack of sleep, day and night upside down, unbalanced diet and other physiological and psychological pressures that cause skin redness and the main reason of uneven skin tone, people can only cover up skin blemishes with makeup, however, the use of makeup aggravates the damage of the skin, resulting in decreased skin elasticity, insufficient water retention, roughness, dullness, enlarged pores, wrinkles, The skin has blemishes and/or an overall dull, sallow, and other aging appearance. In addition, these stresses cause metabolic disorders in the skin, making it impossible for the skin to decompose and discharge metabolic waste in time, and a large amount of waste accumulates in the cells. Accelerates the rate of skin aging, leaving the overall appearance looking listless.
此外,敏感性膚質的人,通常皮膚的表皮較薄,不僅容易脫皮脫屑,皮下微血管也較為明顯,常會受到外部環境、物質的影響,陽光、食物、香味、色素、合成纖維都可能造成刺激,造成毛細血管受刺激,且引發發炎反應。In addition, people with sensitive skin usually have a thinner epidermis, which is not only prone to peeling and desquamation, but also has more obvious subcutaneous microvessels, which are often affected by the external environment and substances, such as sunlight, food, fragrance, pigment, and synthetic fibers. Stimulates the capillaries and triggers an inflammatory response.
為了解決上述問題,本領域的技術人員亟需研發出一種調理肌膚的組合物,以造福有此需求的廣大族群。In order to solve the above problems, those skilled in the art urgently need to develop a composition for conditioning the skin, so as to benefit a large group of people in need.
有鑑於此,本發明之目的是為提供一種果香菊( Chamaemelum nobile)萃取物用於製備調理肌膚的組合物之用途,該果香菊萃取物具有抗發炎、提高超氧化物歧化酶的活性、強化肌膚屏障、降低肌膚的紅斑生成及/或提升肌膚的保濕度。本發明的果香菊萃取物提高肌膚防禦力,減少氧化壓力形成自由基,並且減少肌膚發炎,避免毛細血管受刺激而產生發炎反應,效鎮靜舒緩肌膚、減少肌膚泛紅區域,以提升肌膚穩定性及避免膚色不均。 In view of this, the object of the present invention is to provide an extract of Chamaemelum nobile for preparing a composition for conditioning the skin. The extract of Chamaemelum nobile has anti-inflammation and increases the activity of superoxide dismutase , Strengthen the skin barrier, reduce skin erythema and/or increase skin moisture. The fruity chrysanthemum extract of the present invention improves skin defense, reduces oxidative stress to form free radicals, reduces skin inflammation, prevents capillaries from being stimulated to produce inflammatory reactions, calms and soothes skin, reduces skin redness, and improves skin stability. Sexuality and avoid uneven skin tone.
此外,本發明的果香菊萃取物還強化肌膚屏障,以防止水分散失,使肌膚保持水潤,提供肌膚最佳保護力。In addition, the fruity chrysanthemum extract of the present invention also strengthens the skin barrier to prevent water loss, keeps the skin moisturized, and provides the best protection for the skin.
本發明提供了一種果香菊(Chamaemelum nobile)萃取物用於製備調理肌膚的組合物之用途,其中該果香菊萃取物係以一含水之溶劑進行萃取。The present invention provides a use of chamaemelum nobile extract for preparing a skin conditioning composition, wherein the chamaemelum nobile extract is extracted with a solvent containing water.
在一些實施例中,一種果香菊(Chamaemelum nobile)萃取物用於製備調理肌膚的組合物之用途,其中該果香菊萃取物係以一含水之溶劑進行萃取。In some embodiments, an extract of Chamaemelum nobile is used for preparing a skin conditioning composition, wherein the chamaemelum nobile extract is extracted with an aqueous solvent.
在一些實施例中,該組合物用於抗發炎及/或提高超氧化物歧化酶的活性。In some embodiments, the composition is used for anti-inflammation and/or increasing the activity of superoxide dismutase.
在一些實施例中,該組合物是通過降低活性氧物質及/或降低白血球介素-8 (IL-8)基因的表現量。In some embodiments, the composition works by reducing reactive oxygen species and/or reducing the expression of interleukin-8 (IL-8) gene.
在一些實施例中,該組合物用於強化肌膚屏障。In some embodiments, the composition is used to strengthen the skin barrier.
在一些實施例中,該組合物用於降低肌膚的紅斑生成。In some embodiments, the composition is used to reduce erythema production in the skin.
在一些實施例中,該組合物用於提升肌膚的保濕度。In some embodiments, the composition is used to increase skin hydration.
在一些實施例中,該果香菊萃取物的濃度為至少1重量%。In some embodiments, the concentration of the chrysanthemum extract is at least 1% by weight.
在一些實施例中,該組合物進一步包含一肌膚上可接受之載體。In some embodiments, the composition further comprises a dermatologically acceptable carrier.
在一些實施例中,該組合物之形式為粉末狀、顆粒狀、液狀、膠狀或膏狀之劑型。In some embodiments, the composition is in the form of powder, granule, liquid, jelly or paste.
在一些實施例中,該組合物係進一步製備成保養品組合物、保健食品組合物、或化妝品組合物。In some embodiments, the composition is further prepared into a skin care product composition, a health food composition, or a cosmetic composition.
綜上所述,任一實施例的果香菊萃取物可用於製備調理肌膚的組合物,該果香菊萃取物具有抗發炎、提高超氧化物歧化酶的活性、強化肌膚屏障、降低肌膚的紅斑生成及/或提升肌膚的保濕度。本發明的果香菊萃取物提高肌膚防禦力,減少氧化壓力形成自由基,並且減少肌膚發炎,避免毛細血管受刺激而產生發炎反應,效鎮靜舒緩肌膚、減少肌膚泛紅區域,以提升肌膚穩定性及避免膚色不均。此外,本發明的果香菊萃取物還強化肌膚屏障,以防止水分散失,使肌膚保持水潤,提供肌膚最佳保護力。In summary, the chrysanthemum extract of any embodiment can be used to prepare a composition for conditioning the skin. The chrysanthemum extract has the functions of anti-inflammation, improving the activity of superoxide dismutase, strengthening the skin barrier, reducing skin Erythema formation and/or increase skin hydration. The fruity chrysanthemum extract of the present invention improves skin defense, reduces oxidative stress to form free radicals, reduces skin inflammation, prevents capillaries from being stimulated to produce inflammatory reactions, calms and soothes skin, reduces skin redness, and improves skin stability. Sexuality and avoid uneven skin tone. In addition, the fruity chrysanthemum extract of the present invention also strengthens the skin barrier to prevent water loss, keeps the skin moisturized, and provides the best protection for the skin.
以下將描述本案的部分具體實施態樣。在不背離本案精神下,本案尚可以多種不同形式之態樣來實踐,不應將保護範圍限於說明書所具體陳述的條件。Some specific implementation aspects of this case will be described below. Without departing from the spirit of this case, this case can still be practiced in many different forms, and the scope of protection should not be limited to the conditions specifically stated in the specification.
本案使用Excel軟體進行統計分析。數據以平均值±標準差(SD)表示,各組之間的差異以學生t檢驗(student's t-test)進行分析。In this case, Excel software was used for statistical analysis. Data are expressed as mean ± standard deviation (SD), and differences among groups were analyzed by Student's t-test.
本文中所使用數值為近似值,所有實驗數據皆表示在正負10%的範圍內,最佳為在正負5%的範圍內。The values used in this article are approximate values, and all experimental data are expressed within the range of plus or minus 10%, and the best is within the range of plus or minus 5%.
如本文中所使用,術語“萃取物”係指藉由萃取作用所製備之產物。該萃取物可以溶於溶劑中之溶液形式呈現,或萃取物可為不含或大體上不含溶劑之濃縮物或精華呈現。As used herein, the term "extract" refers to a product prepared by extraction. The extract may be presented as a solution in a solvent, or the extract may be presented as a concentrate or essence free or substantially free of solvent.
果香菊的學名係 Chamaemelum nobile,也稱為羅馬洋甘菊,生長在歐洲、美洲及亞洲等地區。果香菊是為菊科果香菊屬的一種多年生草本植物,有強烈的香味,高可達30公分,莖直立,葉片互生,無柄,二至三回羽狀全裂,末回裂片很狹,頭狀花序單生於莖和長枝頂端,異型花;總苞片具寬膜質邊緣,覆瓦狀排列。花托圓錐形,舌狀花雌性,白色,管狀花兩性,黃色。瘦果無冠狀冠毛。果香菊促進消化、止吐、抗痙攣、輕度鎮靜的作用。果香菊還可用於治療皮膚皸裂、牙齦痛、皮疹、炎症和促進傷口癒合。 The scientific name of fruity chrysanthemum is Chamaemelum nobile , also known as Roman chamomile, which grows in Europe, America and Asia. Fruity Chrysanthemum is a perennial herb of the genus Fruity Chrysanthemum in the family Asteraceae. It has a strong fragrance and can reach a height of 30 cm. The stems are erect, the leaves are alternate, sessile, pinnatifid two to three times, and the last lobes are very large. Narrow, capitulum solitary on the top of stems and long branches, heteromorphic flowers; involucral bracts with broad membranous margins, imbricate. Receptacles conical, ligulate flowers female, white, tubular flowers bisexual, yellow. Achenes without crown hairs. Fruity chrysanthemum promotes digestion, antiemetic, antispasmodic, and mildly sedative. Fructus chamomile can also be used to treat chapped skin, sore gums, rashes, inflammation and to promote wound healing.
如本文所用「果香菊」通常係指植物花朵,其中花朵可包含原始、經乾燥或以其他物理方式加工以利於處理之花朵,其可進一步包含完整、剁碎、切丁、碾磨、研磨或以其他方式經加工以影響原物料之大小及實體完整性之花朵。As used herein, "fruit chrysanthemum" generally refers to the flower of a plant, wherein the flower can include raw, dried or otherwise physically processed to facilitate handling, which can further include whole, chopped, diced, milled, ground Flowers that have been otherwise processed to affect the size and physical integrity of the original material.
在一些實施例中,首先,以人工或機器的方式採摘果香菊的花朵並以清水沖洗備用,將果香菊乾料與水、醇、或醇水混合物之萃取溶劑,萃取溶劑較佳為水,以15-75:1-5之重量比混合,均質後在溶劑中以50-100℃進行萃取0.5-3小時。萃取後冷卻至室溫,將該粗萃取物經由400 mesh之濾網過濾以獲得濾液。最後,將該濾液於45-70℃進行減壓濃縮,最後,將該濾液以噴霧乾燥機進行噴霧乾燥後,得到之該粉劑即為本發明之果香菊萃取物。In some embodiments, firstly, the flowers of Chrysanthemum chrysanthemum are picked manually or by machine and rinsed with clear water for later use. The dry material of chrysanthemum chrysanthemum is extracted with water, alcohol, or a mixture of alcohol and water. The extraction solvent is preferably Water is mixed at a weight ratio of 15-75:1-5, homogenized and then extracted in a solvent at 50-100°C for 0.5-3 hours. After cooling to room temperature after extraction, the crude extract was filtered through a 400 mesh filter to obtain a filtrate. Finally, the filtrate is concentrated under reduced pressure at 45-70° C., and finally, the filtrate is spray-dried by a spray dryer, and the powder obtained is the chrysanthemum chrysanthemum extract of the present invention.
在一些實施例中,由果香菊所榨取的汁液(有時亦稱之為「果香菊萃取物」)可減少黑色素生成、提升肌膚保濕度、減少肌膚皺紋、鎮靜舒緩肌膚及減少肌膚泛紅區域。因此,果香菊萃取物可用於製備抗發炎、提高超氧化物歧化酶的活性、強化肌膚屏障、降低肌膚的紅斑生成及/或提升肌膚的保濕度的肌膚調理的組合物。In some embodiments, the juice extracted from chrysanthemum chrysanthemum (sometimes referred to as chrysanthemum chrysanthemum extract) reduces melanin production, improves skin hydration, reduces skin wrinkles, calms and soothes skin, and reduces skin discoloration. red area. Therefore, the chrysanthemum extract can be used to prepare skin conditioning compositions for anti-inflammation, increasing the activity of superoxide dismutase, strengthening the skin barrier, reducing skin erythema and/or improving skin moisture.
在一些實施例中,含有果香菊萃取物的組合物可為醫藥組合物、保養品組合物、食品組合物、保健食品組合物。In some embodiments, the composition containing the fruit extract can be a pharmaceutical composition, a skin care product composition, a food composition, or a health food composition.
醫藥組合物可利用熟習此技藝者所詳知的技術而被製造成一適合於經腸道地(enterally)、非經腸道地(parenterally)或局部地(topically)投藥的劑型。例如可為:注射品(injection)[例如,無菌的水性溶液(sterile aqueous solution)或分散液(dispersion)]、無菌的粉末(sterile powder)、外部製劑(external preparation)或其他類似物。The pharmaceutical composition can be formulated into a dosage form suitable for enterally, parenterally or topically administered by techniques well known to those skilled in the art. For example, it may be an injection (for example, a sterile aqueous solution or dispersion), a sterile powder, an external preparation, or the like.
醫藥組合物可進一步包含有一被廣泛地使用於藥物製造技術之醫藥上可接受的載劑(pharmaceutically acceptable carrier)。例如,醫藥上可接受的載劑可包含下列一種或多種載劑:乳化劑(emulsifier)、懸浮劑(suspending agent)、分解劑(decomposer)、崩解劑(disintegrating agent)、分散劑(dispersing agent)、黏結劑(binding agent)、賦形劑(excipient)、安定劑(stabilizing agent)、螯合劑(chelating agent)、稀釋劑(diluent)、膠凝劑(gelling agent)、防腐劑(preservative)、潤濕劑(wetting agent)、潤滑劑(lubricant)、吸收延遲劑(absorption delaying agent)、脂質體(liposome)以及類似之物。有關這些載劑的選用與數量是熟習此項技術人員可視情況進行選擇的。The pharmaceutical composition may further contain a pharmaceutically acceptable carrier (pharmaceutically acceptable carrier) which is widely used in pharmaceutical manufacturing technology. For example, a pharmaceutically acceptable carrier may contain one or more of the following: emulsifier, suspending agent, decomposer, disintegrating agent, dispersing agent ), binder (binding agent), excipient (excipient), stabilizer (stabilizing agent), chelating agent (chelating agent), diluent (diluent), gelling agent (gelling agent), preservative (preservative), Wetting agents, lubricants, absorption delaying agents, liposomes, and the like. The selection and quantity of these carriers can be selected by those skilled in the art depending on the circumstances.
在一些實施例中,醫藥上可接受的載劑可包含下列一種溶劑:水、生理鹽水(normal saline)、磷酸鹽緩衝生理鹽水(phosphate buffered saline,PBS)、含有醇的水性溶液(aqueous solution containing alcohol)以及其他任何合適的溶劑。In some embodiments, the pharmaceutically acceptable carrier may comprise one of the following solvents: water, normal saline, phosphate buffered saline (PBS), aqueous solution containing alcohol (aqueous solution containing alcohol) and any other suitable solvent.
在一些實施例中,該醫藥組合物可由下列所述的任一種非經腸道途徑(parenteral routes)來投藥:皮下注射(subcutaneous injection)、表皮內注射(intraepidermal injection)、皮內注射(intradermal injection)以及病灶內注射(intralesional injection)。In some embodiments, the pharmaceutical composition may be administered by any of the following parenteral routes: subcutaneous injection, intraepidermal injection, intradermal injection ) and intralesional injection.
在一些實施例中,醫藥組合物可利用熟習此技藝者所詳知的技術而被製造成一適合於局部地施用於皮膚上的外部製劑(external preparation)。舉例而言,其可為下列所述的任一種,但不限於此:乳劑(emulsion)、凝膠(gel)、軟膏(ointment)、乳霜(cream)、貼片(patch)、擦劑(liniment)、粉末(powder)、氣溶膠(aerosol)、噴霧(spray)、乳液(lotion)、乳漿(serum)、糊劑(paste)、泡沫(foam)、滴劑(drop)、懸浮液(suspension)、油膏(salve)以及繃帶(bandage)。In some embodiments, the pharmaceutical composition may be formulated as an external preparation suitable for topical application to the skin using techniques well known to those skilled in the art. For example, it may be any of the following, but not limited to: emulsion (emulsion), gel (gel), ointment (ointment), cream (cream), patch (patch), liniment ( liniment), powder (powder), aerosol (aerosol), spray (spray), emulsion (lotion), serum (serum), paste (paste), foam (foam), drop (drop), suspension ( suspension), salve, and bandage.
在一些實施例中,外部製劑是藉由將醫藥組合物與一為熟習此項技藝者所詳知的基底(base)相混合而製成。In some embodiments, topical formulations are prepared by mixing the pharmaceutical composition with a base well known to those skilled in the art.
在一些實施例中,該基底可包含下列一種或多種的添加劑(additives):水、醇(alcohols)、甘醇(glycol)、碳氫化合物(hydrocarbons)[諸如石油膠(petroleum, jelly)以及白凡士林(white petrolatum,)]、蠟(wax)[諸如石蠟(paraffin)以及黃蠟(yellow wax)]、保存劑(preserving agents)、抗氧化劑(antioxidants)、界面活性劑(surfactants)、吸收增強劑(absorption enhancers)、安定劑(stabilizing agents)、膠凝劑(gelling agents)[諸如卡波普®974P (carbopol®974P)、微結晶纖維素(microcrystalline cellulose)以及羧基甲基纖維素(carboxymethylcellulose)]、活性劑(active agents)、保濕劑(humectants)、氣味吸收劑(odor absorbers)、香料(fragrances)、pH調整劑(pH adjusting agents)、螯合劑(chelating agents)、乳化劑(emulsifiers)、閉塞劑(occlusive agents)、軟化劑(emollients)、增稠劑(thickeners)、助溶劑(solubilizing agents)、滲透增強劑(penetration enhancers)、抗刺激劑(anti-irritants)、著色劑(colorants)以及推進劑(propellants)等。有關這些添加劑的選用與數量是落在熟習此項技術之人士的專業素養與例行技術範疇內。In some embodiments, the substrate may contain one or more of the following additives: water, alcohols, glycols, hydrocarbons (such as petroleum jelly and white Vaseline (white petrolatum,)], waxes (such as paraffin and yellow wax), preservatives (preserving agents), antioxidants (antioxidants), surfactants (surfactants), absorption enhancers ( absorption enhancers), stabilizing agents, gelling agents (such as carbopol® 974P (carbopol® 974P), microcrystalline cellulose, and carboxymethylcellulose], Active agents, humectants, odor absorbers, fragrances, pH adjusting agents, chelating agents, emulsifiers, occlusive agents (occlusive agents), emollients, thickeners, solubilizing agents, penetration enhancers, anti-irritants, colorants and propellants (propellants) etc. The selection and amounts of these additives are within the professionalism and routine skill of those skilled in the art.
在一些實施例中,保養品中可包含有一被廣泛地使用於保養品製造技術之可接受的佐劑(acceptable adjuvant)。例如,該可接受的佐劑可包含下列一種或多種佐劑:溶劑、膠凝劑、活性劑、防腐劑、抗氧化劑、遮蔽劑(screening agent)、螯合劑、界面活性劑、染色試劑(coloring agent)、增稠劑(thickening agent)、填料(filler)、香料以及氣味吸收劑。可根據實際需求對這些試劑的選用與數量進行合適的調整。In some embodiments, the skin care product may contain an acceptable adjuvant that is widely used in the manufacturing technology of skin care products. For example, the acceptable adjuvant may comprise one or more of the following adjuvants: solvents, gelling agents, active agents, preservatives, antioxidants, screening agents, chelating agents, surfactants, coloring agents agent), thickening agent, filler, fragrance and odor absorber. The selection and quantity of these reagents can be appropriately adjusted according to actual needs.
在一些實施例中,保養品可利用熟習此技藝者所詳知的技術而被製造成一適合於護膚(skincare)或化妝(makeup)的形式,其可為下列中的任一種,但不限於此:水性溶液(aqueous solution)、水-醇溶液(aqueous-alcohol solution)或油性溶液(oily solution)、呈水包油型(oil-in-water type)、油包水型(water-in-oil type)或複合型之乳劑、凝膠、軟膏、乳霜、面膜(mask)、貼片、貼布(pack)、擦劑、粉末、氣溶膠、噴霧、乳液、乳漿、糊劑、泡沫、分散液、滴劑、慕斯(mousse)、防曬油(sunblock)、防曬精華液、防曬乳霜、防曬噴劑、防曬精華、防曬乳霜、防曬噴霧、化妝水(tonic water)、粉底(foundation)、卸妝產品(makeup remover products)、肥皂(soap)以及其他身體清潔產品(body cleansing products)等。In some embodiments, the skin care product can be manufactured into a form suitable for skincare or makeup using techniques well known to those skilled in the art, which can be any of the following, but not limited thereto : Aqueous solution, water-alcohol solution or oily solution, oil-in-water type, water-in-oil type type) or compound emulsion, gel, ointment, cream, mask, patch, pack, liniment, powder, aerosol, spray, lotion, whey, paste, foam, Dispersion, drops, mousse, sunblock, sunscreen essence, sunscreen cream, sunscreen spray, sunscreen essence, sunscreen cream, sunscreen spray, tonic water, foundation ), makeup remover products (makeup remover products), soap (soap) and other body cleansing products (body cleansing products), etc.
在本發明的一實施例中,提供化妝品組合物和個人護理產品,其包含能夠顯示變色的著色組合物,例如包含在載體和苯乙烯-馬來酸酐共聚物中的顏料顆粒的顏料研磨物。化妝品組合物可以是給人體皮膚提供色彩的任意化妝品或個人護理產品形式。例如,化妝品組合物可以是、但不限於唇膏、唇彩、保濕唇膏、指甲油、粉底、撲面粉、底粉、遮瑕膏、腮紅、眼影、眼線器、睫毛膏或古銅色化妝品。個人護理產品可以是給人體皮膚賦予色彩的任意適合的形式。例如,個人護理產品可以包括日霜或洗劑、晚霜或洗劑、防曬露、霜或油和其它SPF產品、增濕劑、油膏、軟膏、凝膠、體乳、人造褐變組合物、脫毛等。In one embodiment of the present invention, cosmetic compositions and personal care products are provided comprising a coloring composition capable of exhibiting color change, such as a pigment grind comprising pigment particles in a carrier and styrene-maleic anhydride copolymer. The cosmetic composition may be in the form of any cosmetic or personal care product that provides color to human skin. For example, the cosmetic composition can be, but is not limited to, lipstick, lip gloss, moisturizing lipstick, nail polish, foundation, face powder, primer, concealer, blush, eye shadow, eyeliner, mascara, or bronzer. The personal care product may be in any suitable form for imparting color to human skin. For example, personal care products may include day creams or lotions, night creams or lotions, sunscreen lotions, creams or oils and other SPF products, moisturizers, salves, ointments, gels, body milks, artificial browning compositions, hair removal etc.
本發明的化妝品組合物和個人護理產品施用於人皮膚系統,包括皮膚、唇、指甲、毛髮和其它角質表面。本文所用的術語“角質表面”是指包含角蛋白的人皮膚系統的部分,包括但不限於哺乳動物、優選人類的皮膚、唇、毛髮(包括頭皮、睫毛、眉毛、面毛和體毛)和指甲(腳趾甲、指甲、甲上皮等)。The cosmetic compositions and personal care products of the present invention are applied to the human dermal system, including skin, lips, nails, hair and other keratinous surfaces. The term "keratinous surface" as used herein refers to the portion of the human skin system comprising keratin, including but not limited to mammalian, preferably human, skin, lips, hair (including scalp, eyelashes, eyebrows, facial and body hair) and Nails (toenails, fingernails, nail epithelium, etc.).
能夠顯示變色的化妝品或個人護理產品施用於皮膚的任意區域且優選面部、頸部、手、足或身體的另外的區域上,例如壁、腿和背部。Cosmetic or personal care products capable of exhibiting discoloration are applied to any area of the skin and preferably the face, neck, hands, feet or other areas of the body, such as the walls, legs and back.
在一些實施例中,保養品亦可與下列中之已知活性的外用劑(external use agents)的一種或多種一起合併使用:美白劑(whitening agents)[諸如維生素A酸(tretinoin)、兒茶素(catechin)、麴酸、熊果苷以及維生素C]、保濕劑、殺菌劑(bactericides)、紫外線吸收劑(ultraviolet absorbers)、植物萃取物(plant extracts)[諸如蘆薈萃取物(aloe extract)]、皮膚營養劑(skin nutrients)、麻醉劑(anesthetics)、抗痘劑(anti-acne agents)、止癢劑(antipruritics)、止痛劑(analgesics)、抗皮膚炎劑(antidermatitis agents)、抗過角化劑(antihyperkeratolytic agents)、抗乾皮膚劑(anti-dry skin agents)、抗汗劑(antipsoriatic agents)、抗老化劑(antiaging agents)、抗皺劑(antiwrinkle agents)、抗皮脂溢出劑(antiseborrheic agents)、傷口治療劑(wound-healing agents)、皮質類固醇(corticosteroids)以及激素(hormones)。有關這些外用劑的選用與數量是落在熟習此項技術之人士的專業素養與例行技術範疇內。In some embodiments, skin care products may also be used in combination with one or more of the following known active external use agents: whitening agents (such as tretinoin, catechin catechin, kojic acid, arbutin, and vitamin C], humectants, bactericides, ultraviolet absorbers, plant extracts (such as aloe extract) , skin nutrients, anesthetics, anti-acne agents, antipruritics, analgesics, antidermatitis agents, anti-keratosis antihyperkeratolytic agents, anti-dry skin agents, antipsoriatic agents, antiaging agents, antiwrinkle agents, antiseborrheic agents, Wound-healing agents, corticosteroids, and hormones. The selection and amount of these topical agents are within the professionalism and routine skill of those skilled in the art.
在一些實施例中,醫藥組合物可被當作食品添加物(food additive),藉由習知方法於原料製備時添加,或是於食品的製作過程中添加,而與任一種可食性材料配製成供人類與非人類動物攝食的食品產品。In some embodiments, the pharmaceutical composition can be used as a food additive, which can be added during the preparation of raw materials or during the production of food by conventional methods, and can be formulated with any edible material Prepared as a food product intended for consumption by humans and non-human animals.
在一些實施例中,食品的種類可為但不限於:飲料(beverages)、發酵食品(fermented foods)、烘培產品(bakery products)、健康食品(health foods)以及膳食補充品(dietary supplements)。該食品包括但不限於飲品、果凍、乳製品、代餐包等。In some embodiments, the types of food may be but not limited to: beverages, fermented foods, bakery products, health foods and dietary supplements. The food includes but is not limited to beverages, jellies, dairy products, meal replacement packs, etc.
下列多個實施例中的實驗步驟若無特別敘明,即在室溫(25±5℃)、常壓(1atm)下進行。Unless otherwise specified, the experimental procedures in the following examples are carried out at room temperature (25±5° C.) and normal pressure (1 atm).
[實施例1]:果香菊萃取物的製備方法[Example 1]: the preparation method of fruity chrysanthemum extract
在此實施例中,首先,以人工或機器的方式採摘果香菊的花朵並以清水沖洗備用,將果香菊乾料與水、醇、或醇水混合物之萃取溶劑,萃取溶劑較佳為水,果香菊乾料與萃取溶劑以1:15之重量比混合,均質後在溶劑中以85℃進行萃取1小時。萃取後冷卻至室溫,將該粗萃取物經由400 mesh之濾網過濾以獲得濾液。最後,將該濾液於60℃±5℃進行減壓濃縮,最後,將該濾液以噴霧乾燥機進行噴霧乾燥後,得到之該粉劑即為本發明之果香菊萃取物。In this embodiment, at first, the flowers of Chrysanthemum chrysanthemum are picked manually or by machine and rinsed with clear water for later use. The dry material of chrysanthemum chrysanthemum is extracted with water, alcohol, or a mixture of alcohol and water. The extraction solvent is preferably Mix water, dried chrysanthemum and extraction solvent at a weight ratio of 1:15, and extract in the solvent at 85°C for 1 hour after homogenization. After cooling to room temperature after extraction, the crude extract was filtered through a 400 mesh filter to obtain a filtrate. Finally, the filtrate is concentrated under reduced pressure at 60°C±5°C, and finally, the filtrate is spray-dried by a spray dryer, and the powder obtained is the chrysanthemum chrysanthemum extract of the present invention.
[實施例2]:果香菊萃取物的總黃酮含量檢測[Example 2]: detection of total flavonoid content of fruity chrysanthemum extract
總黃酮含量檢測的分析方法主要是參考Zhishen Jia et al.,(1999),Food Chemistry,64:555-559所述方法並加以修飾,取200μL之適當稀釋後的樣品加入1000μL之H
2O,混合均勻後,加入200μL之5%檸檬酸鈉(Sodium nitrite),靜置6分鐘後加入200μL之10%硝酸鋁(Aluminum nitrate),再靜置6分鐘。最後加入2mL之4%氫氧化鈉(Sodium hydroxide)與1.4mL之H
2O,混合均勻,於500nm下進行分析,以芸香素(Rutin)為標準品繪製標準曲線。在本實施例中,實施例1之果香菊萃取物的總黃酮的含量為至少3.0 mg/mL。
The analytical method for detecting the content of total flavonoids is mainly based on the method described by Zhishen Jia et al., (1999), Food Chemistry, 64: 555-559 and modified. Take 200 μL of an appropriately diluted sample and add 1000 μL of H 2 O. After mixing evenly, add 200 μL of 5% sodium citrate (Sodium nitrite), let it stand for 6 minutes, then add 200 μL of 10% aluminum nitrate (Aluminum nitrate), and let it stand for another 6 minutes. Finally, 2 mL of 4% sodium hydroxide (Sodium hydroxide) and 1.4 mL of H 2 O were added, mixed uniformly, analyzed at 500 nm, and a standard curve was drawn with Rutin as a standard. In this embodiment, the total flavonoid content of the chrysanthemum extract of
[實施例3]:細胞實驗-果香菊萃取物降低活性氧物質[Example 3]: Cell experiment - fruity chrysanthemum extract reduces active oxygen species
於此,以螢光顯微鏡避光觀察人類皮膚纖維母細胞CCD-966sk細胞內綠螢光表現之程度,以測定經果香菊萃取物處理後,其活性氧物質含量的變化。Herein, the degree of green fluorescence in human dermal fibroblasts CCD-966sk cells was observed with a fluorescent microscope in the dark, so as to determine the changes in the content of reactive oxygen species after being treated with the extract of chrysanthemum chrysanthemum.
材料與儀器
1. 細胞株:人類皮膚纖維母細胞(Human skin fibroblast)CCD-966sk(生物資源保存及研究中心(BCRC),No. 60153)。
2. 培養基:含有10 vol% FBS(fetal bovine serum,購自Gibco)之基礎培養基。其中,基礎培養基是由Eagle’s最低限度基本培養基(Eagle’s minimum essential medium(MEM),購自Gibco,產品編號15188-319)額外添加成分使其含有1 mM 丙酮酸鈉(sodium pyruvate,購自Gibco)、1.5 g/L 碳酸氫鈉(sodium bicarbonate,購自Sigma)所配製而成。
3. 磷酸緩衝鹽溶液(PBS溶液):購自Gibco,產品編號10437-028。
4. DCFH-DA溶液:將二氯二氫螢光素二乙酸酯(2,7-dichloro-dihydro-fluorescein diacetate,DCFH-DA;產品編號SI-D6883,購自Sigma)溶於二甲基亞碸(dimethyl sulfoxide,DMSO,購自Sigma,產品編號SI-D6883-50MG)以配製成5 mg/ml 的DCFH-DA溶液。
5. 雙氧水(H
2O
2):購自Sigma-Aldrich,產品型號95299-1L。
6. 胰蛋白酶(Trypsin-EDTA):10X Trypsin-EDTA(購自Gibco)以1X PBS溶液稀釋10倍。
7. 倒立式顯微鏡搭載照相系統 (ZEISS,Cat. Vert.A1)
8. 果香菊萃取物:此實驗所使用的果香菊萃取物係以實施例1之方式製備而成。
Materials and
實驗步驟Experimental procedure
實驗將會分為實驗組、控制組(未添加果香菊萃取物、亦無經過雙氧水處理的組別)、以及對照組(未添加果香菊萃取物,但經雙氧水處理的組別)三組進行,各組分別進行三重複試驗: 1. 將CCD-966sk細胞以每孔1×10 5個的方式,接種於每孔含2ml培養基之6孔培養盤中。 2. 將培養盤置於5%CO 2、37℃下,培養24小時。 3. 移除培養基。 4. 加入2 mL實驗培養基至培養盤的各孔中,並於37 ℃下培養1小時。 實驗組的實驗培養基為添加有5 μL的實施例1得到的果香菊萃取物之2 mL細胞培養基(即果香菊萃取物佔細胞培養基的體積百分比為0.125%)。 控制組的實驗培養基為單純的2 mL細胞培養基(即不含果香菊萃取物)。 對照組的實驗培養基為單純的2 mL細胞培養基(即不含果香菊萃取物)。 5. 添加DCFH-DA溶液使其於培養基的濃度為10μg/ml,於37℃下反應40分鐘。 6. 於DCFH-DA處理後,於實驗組的實驗培養基以及對照組的實驗培養基分別加入H 2O 2,並於37 ℃下反應1小時。具體來說,35% wt的雙氧水先稀釋成100 mM(將10 μL的雙氧水加入990 μL的二次蒸餾水),再取20 μL的100 mM的雙氧水加入 2 mL的細胞培養盤中。 7. 反應後,每孔以1 ml PBS溶液清洗細胞2次。 8. 以Hoechst 33342 (1:20000 稀釋)進行染色,避光作用5分鐘。 9. 以螢光顯微鏡避光觀察各組細胞,並拍攝照片以紀錄細胞內綠螢光表現之程度。相關圖式如圖1。 The experiment will be divided into the experimental group, the control group (the group that did not add the extract of chrysanthemum chrysanthemum, and the group that was not treated with hydrogen peroxide), and the control group (the group that did not add the extract of chrysanthemum chrysanthemum, but was treated with hydrogen peroxide) The experiment was carried out in groups, and each group carried out three repeated experiments: 1. CCD-966sk cells were inoculated in a 6-well culture plate containing 2ml of medium in each well at a rate of 1×10 5 per well. 2. Place the culture plate in 5% CO 2 at 37°C and incubate for 24 hours. 3. Remove medium. 4. Add 2 mL of assay medium to each well of the culture plate and incubate at 37°C for 1 hour. The experimental medium of the experimental group was 2 mL of cell culture medium supplemented with 5 μL of the fruity chrysanthemum extract obtained in Example 1 (that is, the volume percentage of the fruity chrysanthemum extract to the cell culture medium was 0.125%). The experimental medium of the control group was simple 2 mL cell culture medium (that is, no fruit extract). The experimental medium of the control group was a simple 2 mL cell culture medium (that is, no fruit extract). 5. Add DCFH-DA solution so that the concentration in the culture medium is 10 μg/ml, and react at 37°C for 40 minutes. 6. After DCFH-DA treatment, add H 2 O 2 to the experimental medium of the experimental group and the experimental medium of the control group respectively, and react at 37°C for 1 hour. Specifically, 35% wt hydrogen peroxide was first diluted to 100 mM (add 10 μL of hydrogen peroxide to 990 μL of double distilled water), and then 20 μL of 100 mM hydrogen peroxide was added to a 2 mL cell culture dish. 7. After the reaction, wash the cells twice with 1 ml PBS solution per well. 8. Stain with Hoechst 33342 (diluted 1:20000) for 5 minutes in the dark. 9. Use a fluorescent microscope to observe each group of cells in the dark, and take pictures to record the degree of green fluorescence in the cells. The relevant diagram is shown in Figure 1.
實驗結果Experimental results
如圖1所示,控制組的ROS含量為1.0,由空白控制組、對照組的結果可知,在經過雙氧水處理後後,具有高ROS表現(高螢光表現)的細胞比例大幅增加至13.9倍,顯示雙氧水確實會導致細胞內產生活性氧物質,進而對纖維母細胞產生後續傷害。另一方面,根據對照組以及實驗組的結果可知,當細胞經過果香菊萃取物處理後,具有高ROS表現的細胞比例相較於對照組明顯減少至1.4倍。顯示果香菊萃取物可有效減少活性氧物質在細胞內的產生或累積,可作為一種活性氧物質清除劑。亦即,果香菊萃取物可透過降低細胞內活性氧物質含量,減少細胞受到雙氧水等強氧化物質等所導致的氧化傷害。As shown in Figure 1, the ROS content of the control group was 1.0. From the results of the blank control group and the control group, it can be seen that after hydrogen peroxide treatment, the proportion of cells with high ROS expression (high fluorescence expression) increased significantly to 13.9 times, showing that Hydrogen peroxide does lead to intracellular production of reactive oxygen species with subsequent damage to fibroblasts. On the other hand, according to the results of the control group and the experimental group, it can be seen that when the cells were treated with the fruit extract, the proportion of cells with high ROS expression was significantly reduced to 1.4 times compared with the control group. It is shown that the extract of chrysanthemum chrysanthemum can effectively reduce the production or accumulation of active oxygen species in cells, and can be used as a scavenger of active oxygen species. In other words, the fruit extract can reduce the oxidative damage to cells caused by strong oxidizing substances such as hydrogen peroxide by reducing the content of active oxygen species in cells.
[實施例4]:細胞實驗-果香菊萃取物提升超氧化物歧化酶的活性[Example 4]: Cell experiment - fruity chrysanthemum extract enhances the activity of superoxide dismutase
人類皮膚纖維母細胞購自台灣生物資源保存及研究中心(Bioresource Collection and Research Center,BCRC),編號BCRC 60153。將該細胞培養於添加10%胎牛血清(fetal bovine serum,FBS)(GIBCO公司,編號10438-026,美國)、0.1mM非必需胺基酸、1.5g/L碳酸氫鈉(Sigma公司,編號S5761,美國)、1mM丙酮酸鈉(GIBCO公司,編號11360-070,美國)的最低必需培養液(Minimum essential medium,MEM)(Eagle)(配於厄爾平衡鹽溶液(Earle's Balanced Salt Solution,Earle's BSS))(GIBCO公司,編號41500-034,美國)。Human dermal fibroblasts were purchased from Taiwan Bioresource Collection and Research Center (BCRC), number BCRC 60153. The cells were cultured in a medium supplemented with 10% fetal bovine serum (FBS) (GIBCO Company, No. 10438-026, U.S.), 0.1 mM non-essential amino acids, 1.5 g/L sodium bicarbonate (Sigma Company, No. S5761, the United States), 1mM sodium pyruvate (GIBCO company, No. 11360-070, the United States) minimum essential medium (Minimum essential medium, MEM) (Eagle) (mixed with Earle's Balanced Salt Solution (Earle's Balanced Salt Solution, Earle's BSS)) (GIBCO Corporation, No. 41500-034, USA).
在含有2mL上述培養基的6孔培養盤中每個孔洞接種2×105個人類皮膚纖維母細胞,然後於37℃培養隔夜。在移除培養基之後,將細胞分成2組,其中包括1個對照組及1個實驗組。將0.5mg/mL 果香菊萃取物及1mM H2O2添加至實驗組的細胞中並於37℃作用6小時,至於對照組的細胞則不做任何處理。接著,將各組細胞於37℃下培養24小時。之後,移除培養基並以1X PBS(Gibco)清洗1次,然後加入胰蛋白酶(trypsin)來處理細胞3分鐘,繼而用培養基停止胰蛋白酶活性並轉移至1.5mL離心管中。接著,以300xg離心5分鐘,然後以PBS清洗,接而以300xg離心5分鐘。之後,加入30μL之1X細胞萃取緩衝液,然後將細胞懸浮液在冰上反應30分鐘,每10分鐘渦旋(vortex)混合。接著,將破碎的細胞懸浮液在4℃以10,000xg離心10分鐘以除去不溶物質,然後將25μL的1X SOD緩衝液加入活性對照孔的底部。之後,添加25μL製備的SOD標準品(S1~S7,參見表1)。Inoculate 2×105 human dermal fibroblasts into each well of a 6-well culture plate containing 2 mL of the above medium, and then culture overnight at 37°C. After removing the medium, the cells were divided into 2 groups, including 1 control group and 1 experimental group. Add 0.5mg/mL chrysanthemum extract and 1mM H2O2 to the cells of the experimental group and act at 37°C for 6 hours, while the cells of the control group are not treated. Next, each group of cells was cultured at 37° C. for 24 hours. Afterwards, the medium was removed and washed once with 1X PBS (Gibco), and then trypsin was added to treat the cells for 3 minutes, followed by medium to stop trypsin activity and transferred to a 1.5 mL centrifuge tube. Next, centrifuge at 300xg for 5 minutes, then wash with PBS, and then centrifuge at 300xg for 5 minutes. Afterwards, 30 μL of 1X cell extraction buffer was added, and then the cell suspension was reacted on ice for 30 minutes, vortexed every 10 minutes to mix. Next, the broken cell suspension was centrifuged at 10,000 xg for 10 minutes at 4°C to remove insoluble material, and then 25 μL of 1X SOD buffer was added to the bottom of the active control well. After that, add 25 μL of prepared SOD standards (S1~S7, see Table 1).
表1
:
之後,將25 μL的樣品加入適當孔的底部,然後對每孔添加150 μL的Master Mix。接著,使用多道移液器(multichannel pipet)向所有孔中加入25 μL之1X黃嘌呤溶液(Xanthine solution)以起始反應,然後立即將培養盤轉移至微量滴定盤讀取儀(microtiter plate reader),並在室溫下每分鐘以450 nm讀取吸光值,持續10分鐘。Afterwards, 25 μL of the sample was added to the bottom of the appropriate well, followed by the addition of 150 μL of Master Mix to each well. Next, 25 μL of 1X Xanthine solution was added to all wells using a multichannel pipet to initiate the reaction, and the plate was immediately transferred to a microtiter plate reader. ), and read the absorbance at 450 nm every minute for 10 minutes at room temperature.
各組之間的統計學顯著差異是藉由史徒登氏t-檢定來決定。本實施例的結果顯示於圖2。Statistically significant differences between groups were determined by Student's t-test. The results of this example are shown in FIG. 2 .
圖2是本發明果香菊萃取物在提升皮膚纖維母細胞之SOD活性上的功效之數據圖。由圖2可見,與對照組相較之下,實驗組的SOD活性有顯著的提升(與對照組比較,實驗組的SOD活性提升約117.5 %)。本實施例的結果顯示,本發明果香菊萃取物具有提升皮膚纖維母細胞之SOD活性的功效。Fig. 2 is a data graph showing the efficacy of the extract of chrysanthemum chrysanthemum of the present invention in enhancing the SOD activity of dermal fibroblasts. As can be seen from Figure 2, compared with the control group, the SOD activity of the experimental group was significantly improved (compared with the control group, the SOD activity of the experimental group increased by about 117.5%). The results of this example show that the extract of Chrysanthemum chrysanthemum of the present invention has the effect of enhancing the SOD activity of dermal fibroblasts.
[實施例5]:抗皮膚發炎的效用評估[Example 5]: Efficacy Evaluation of Anti-Skin Inflammation
於此,以人類CXCL8/IL8之ELISA分析套組、紫外線照射室配合酵素免疫分析儀(ELISA reader),測定人類初代皮膚角質細胞HPEK-50經果香菊萃取物處理後,其細胞中抗發炎因子的變化。Here, the human CXCL8/IL8 ELISA analysis kit, ultraviolet radiation chamber and enzyme immunoassay analyzer (ELISA reader) were used to measure the anti-inflammatory effect in the cells of primary human skin keratinocytes HPEK-50 treated with chrysanthemum extract. factor changes.
材料與儀器
1. 細胞株:人類初代皮膚角質細胞(Human primary epidermal keratinocytes)HPEKp(CELLnTEC公司(瑞士),HPEK-50)。
2. 培養基:無血清之角質細胞培養液(keratinocyte-SFM) (Gibco公司,美國,編號為#10724-011)。
3. 紫外線照射室(Vilber公司)。
4. 人類IL8之ELISA分析套組:購自R&D systems,型號 D8000CD8000C,此套組中包含以下試劑:
(1) 捕獲抗體(capture antibody)
(2) 偵測抗體(detection antibody)
(3) 鏈黴親和素-HRP(Streptavidin-HRP)
(4) 清洗溶液(Washing buffeer):為磷酸緩衝鹽溶液(PBS溶液)
(5) 受質溶液(四甲基聯苯胺 tetramethyl-benzidine,TMB)
(6) 終止溶液(stop solution)
5. 酵素免疫分析儀(ELISA reader),購自BioTek公司。
6. substrate solution (R&D systems)。
7. H
2SO
4,購自Sigma-Aldrich公司。
8. 細胞培養箱,購自Astec公司。
9. 振盪器(shaker),購自GenePure公司。
10. 果香菊萃取物:此實驗中所使用的果香菊萃取物是透過如上實施例1所獲得。
Materials and
檢測目的Detection purpose
已知介白素8(interleukin-8,IL-8)為一種細胞激素,具有促進發炎反應產生之功能,當發炎反應產生時,因為發炎組織會釋放出特定的細胞激素以吸引特定功能的細胞至特定的組織,IL-8在中性白血球的趨化作用上扮演重要的角色。Interleukin 8 (interleukin-8, IL-8) is known as a cytokine, which has the function of promoting inflammatory response. When an inflammatory response occurs, the inflamed tissue will release specific cytokines to attract cells with specific functions IL-8 plays an important role in the chemotaxis of neutrophils to specific tissues.
紫外光中的UVB具高能量,能穿透至皮膚表皮導致皮膚曬傷,並造成皮膚發炎及紅腫。UVB in ultraviolet light has high energy, which can penetrate into the epidermis of the skin and cause sunburn, and cause skin inflammation and redness.
實驗步驟Experimental procedure
實驗將會分為1個控制組、1個對照組(UVB照射)、1個實驗組(UVB照射及添加果香菊萃取物)共三組進行,各組分別進行三重複試驗: 1. 將人類初代皮膚角質細胞以每孔5×10 4個的方式,接種於每孔培養於每孔含0.5ml培養基之24孔培養盤中。 2. 將培養盤置於細胞培養箱,於37℃下培養24小時後,移除培養基。 3. 使用紫外線照射室,以300mJ/cm 2的強度對對照組、實驗組,誘發發炎反應,並同時將0.125 mg/mL 果香菊萃取物分別添加至實驗組的細胞中,至於控制組的細胞則不做任何處理(不照UVB,亦不添加果香菊萃取物)。 4. 各組細胞在37℃下培養24小時後,吸取120μL的細胞培養上清液做為樣品,使用人類IL8之ELISA分析套組對此些樣品中IL8進行分析。 5. 首先,以PBS稀釋捕獲抗體(capture antibody),將經稀釋的捕獲抗體以每孔100μL的量塗佈至96孔微盤底部,並於4℃作用過夜。 6. 接著,以200μL的清洗緩衝液(含0.05%吐溫20(Tween 20),製備於PBS)清洗96孔微盤3次。 7. 以300μL的阻斷緩衝液(Block buffer)(1% BSA製備於PBS)進行阻斷,接著於37℃下作用3小時。 8. 之後,以200μL的清洗緩衝液清洗96孔微盤3次。 9. 添加100μL的樣品及標準品(配置於稀釋液(0.1% BSA配於PBS)),接而於37℃下與捕捉抗體進行結合2小時。 10. 接著,以200μL的清洗緩衝液清洗96孔微盤3次。 11. 然後添加100μL的偵測抗體(detection antibody),接而於37℃下偵測捕捉抗體2小時。 12. 之後,以200μL的清洗緩衝液清洗96孔微盤3次。 13. 添加100μL的鏈黴親和素-HRP(Streptavidin-HRP),接而於室溫下作用20分鐘。 14. 之後,以200μL的清洗緩衝液清洗96孔微盤3次。 15. 然後添加100μL的受質溶液(substrate solution)(R&D systems),於室溫下作用20分鐘。 16. 接著,添加50μL的終止溶液(stop solution)來中止反應,最後以酵素免疫分析儀測量其450nm之吸光值。再以Excel軟體進行統計分析。 The experiment will be divided into three groups: 1 control group, 1 control group (UVB irradiation), and 1 experimental group (UVB irradiation and adding chrysanthemum chrysanthemum extract), and each group will carry out three repeated experiments: 1. Human primary skin keratinocytes were inoculated at 5×10 4 per well and cultured in a 24-well culture plate containing 0.5ml medium per well. 2. Place the culture plate in the cell culture incubator, culture at 37°C for 24 hours, then remove the culture medium. 3. Using the ultraviolet irradiation room, with the intensity of 300mJ/ cm2 , the control group and the experimental group were induced to inflammatory response, and at the same time, 0.125 mg/mL fruit extract was added to the cells of the experimental group respectively, as for the control group The cells were left without any treatment (no UVB exposure, and no chrysanthemum extract). 4. After each group of cells was cultured at 37°C for 24 hours, 120 μL of cell culture supernatant was taken as a sample, and IL8 in these samples was analyzed using the human IL8 ELISA analysis kit. 5. First, dilute the capture antibody with PBS, apply the diluted capture antibody to the bottom of a 96-well microplate at an amount of 100 μL per well, and react overnight at 4°C. 6. Next, wash the 96-well microplate three times with 200 μL of washing buffer (containing 0.05% Tween 20 (Tween 20), prepared in PBS). 7. Block with 300 μL of blocking buffer (Block buffer) (1% BSA prepared in PBS), and then act at 37°C for 3 hours. 8. Afterwards, wash the 96-well microplate 3 times with 200 μL of washing buffer. 9. Add 100 μL of samples and standards (configured in diluent (0.1% BSA in PBS)), and then bind to the capture antibody at 37°C for 2 hours. 10. Next, wash the 96-well microplate 3 times with 200 μL of washing buffer. 11. Then add 100 μL of detection antibody, and then detect the capture antibody at 37°C for 2 hours. 12. Afterwards, wash the 96-well microplate 3 times with 200 μL of washing buffer. 13. Add 100 μL of streptavidin-HRP (Streptavidin-HRP), then react at room temperature for 20 minutes. 14. After that, wash the 96-well microplate 3 times with 200 μL of washing buffer. 15. Then add 100 μL of substrate solution (R&D systems) for 20 minutes at room temperature. 16. Next, add 50 μL of stop solution (stop solution) to stop the reaction, and finally measure its absorbance at 450 nm with an enzyme immunoassay analyzer. Statistical analysis was performed with Excel software.
各組之間的統計學顯著差異是藉由史徒登氏t-試驗(Student’s t-test)來決定。本實施例的結果顯示於圖3。Statistically significant differences between groups were determined by Student's t-test. The results of this example are shown in FIG. 3 .
圖3是本發明果香菊萃取物在抗皮膚發炎上的功效之數據圖。由圖3可見,與對照組相較之下,UVB組的IL-8產生量有顯著的提升至約115.2%,這表示UVB會對人類初代皮膚角質細胞產生發炎反應。而與UVB組相較之下,實驗組的IL-8產生量降低105.9%。本實施例的結果顯示,本發明果香菊萃取物具有抗皮膚發炎的功效,可使皮膚保護力升高。Fig. 3 is a data graph of the anti-inflammatory effect of the chrysanthemum extract of the present invention on skin. It can be seen from Figure 3 that compared with the control group, the IL-8 production in the UVB group was significantly increased to about 115.2%, which indicates that UVB can cause an inflammatory response to human primary skin keratinocytes. Compared with the UVB group, the production of IL-8 in the experimental group decreased by 105.9%. The results of this example show that the extract of the chrysanthemum chrysanthemum of the present invention has the effect of anti-inflammation of the skin, and can increase the protective power of the skin.
而為證實果香菊萃取物對於人體的功效,亦進行相關人體實驗如下。In order to confirm the effect of the fruit extract on the human body, relevant human experiments are also carried out as follows.
[實施例6]:人體實驗-肌膚的保濕度檢測[Example 6]: human body experiment-skin moisturizing degree detection
使用樣品:含果香菊萃取物的精華液的面膜(以面膜布浸泡於如下所述的含有果香菊萃取物的精華液中,使其充分吸收後製得)以及安慰劑面膜(以相同型號的面膜布浸泡於與前述精華液中,但其中的果香菊萃取物替換成等重的水),再使其充分吸收後製得。於此實施例中,各面膜係含25mL的對應精華液。Samples used: facial mask containing the essence of chrysanthemum extract (made by soaking the mask cloth in the essence solution containing chrysanthemum extract as described below to make it fully absorbed) and placebo mask (made with the same The mask cloth of the model is soaked in the aforementioned essence, but the chrysanthemum extract in it is replaced with water of the same weight), and then it is fully absorbed. In this example, each facial mask contains 25mL of the corresponding essence.
含有果香菊萃取物的精華液成分:三仙膠 0.2wt%、馨鮮酮 0.6wt%、1,3-丁二醇 5wt%、三乙醇胺 0.1wt%、己二醇 0.6wt%、果香菊萃取物0.5wt%,其餘為水。其中,此果香菊萃取物係由實施例1之製備方式而得。Essence ingredients containing fruity chrysanthemum extract: 0.2wt% sanxian gum, 0.6wt% xinxinone,
受試者人數:7位20-55歲之男性及女性受試者。Number of subjects: 7 male and female subjects aged 20-55.
實驗方式:受試者將全臉清潔後,依據不同檢測項目,使用對應的儀器及測量方式,紀錄左右半臉於使用面膜前之數值、或拍攝使用前的照片。而後,針對瞬效試驗,分別將果香菊萃取物面膜以及安慰劑面膜貼敷於受試者的右半臉及左半臉上,經15分鐘後取下,再以指腹稍加按摩左半臉與右半臉。待面膜移除約5分鐘後,再以對應的儀器及測量方式,進行測量或拍攝照片,再與自身原左半臉、或右半臉之數值進行對照(使用前與使用後欲進行檢測時,受試者所在的測試區域的溫度與濕度為一致,以減少外界的溫濕度等因素會對肌膚所造成的影響)。針對長效試驗,對受試者以前述方法連續使用4週之前及之後進行檢測。Experimental method: After the subject cleans the whole face, according to different test items, use the corresponding instrument and measurement method to record the values of the left and right half of the face before using the mask, or take a photo before using it. Then, for the instant test, the fruity chrysanthemum extract mask and the placebo mask were applied to the right half of the face and the left half of the face of the subject respectively, and they were removed after 15 minutes, and then the left half was massaged with the fingertips. Half face and right half face. After the mask is removed for about 5 minutes, use the corresponding instrument and measurement method to measure or take a photo, and then compare it with the value of your original left half face or right half face (when you want to test before and after use) , the temperature and humidity of the test area where the subject is located are consistent to reduce the impact of external temperature and humidity on the skin). For the long-acting test, the test subjects were tested before and after continuous use for 4 weeks by the aforementioned method.
使用購自德國Courage+Khazaka electronic公司之肌膚含水量檢測探頭Corneometer ®CM825 (C+K Multi Probe Adapter System, Germany) 對同一受試者在使用面膜前與後的面部肌膚進行檢測。該檢測探頭係基於電容的原理進行測量。當水分含量發生變化時,肌膚的電容值亦發生變化,故可通過測定肌膚電容值,分析肌膚表面的含水量。 The skin moisture content detection probe Corneometer ® CM825 (C+K Multi Probe Adapter System, Germany) purchased from Courage+Khazaka electronic in Germany was used to detect the facial skin of the same subject before and after using the mask. The detection probe is based on the principle of capacitance to measure. When the water content changes, the capacitance value of the skin also changes, so the water content of the skin surface can be analyzed by measuring the skin capacitance value.
請參閱圖4A。該圖係受試者在使用含本案果香菊萃取物之面膜前與後的瞬效(15分鐘)的肌膚含水量,以受試者使用前的含水量為100%。其中,受試者使用後的肌膚含水量增加至約119.5%,果香菊萃取物能夠立即提供良好的保濕功效,提升肌膚的保濕度,使肌膚不再因乾燥而變成粗糙。See Figure 4A. The figure shows the instant (15 minutes) moisture content of the skin of the subject before and after using the mask containing the chrysanthemum chrysanthemum extract, and the moisture content before use is taken as 100%. Among them, the water content of the subjects' skin increased to about 119.5% after use, and the extract of chrysanthemum chrysanthemum can immediately provide good moisturizing effect, improve the moisturizing degree of the skin, and make the skin no longer rough due to dryness.
請參閱圖4B。該圖係受試者在使用含本案果香菊萃取物之面膜前與後的長效(4週)的肌膚含水量,以受試者使用前的含水量為100%。其中,受試者使用後的肌膚含水量增加至約110.7%,果香菊萃取物能夠長期提供良好的保濕功效,提升肌膚的保濕度,使肌膚不再因乾燥而變成粗糙。See Figure 4B. The figure is the long-term (4 weeks) skin moisture content of the subject before and after using the mask containing the extract of the fruit chrysanthemum in this case, and the moisture content before the subject's use is 100%. Among them, the water content of the subjects' skin increased to about 110.7% after use, and the fruity chrysanthemum extract can provide good moisturizing effect for a long time, improve the moisturizing degree of the skin, and make the skin no longer rough due to dryness.
[實施例7]:人體實驗-肌膚泛紅檢測[Example 7]: Human experiment - skin redness detection
使用美國Canfield所販售之VISIA全臉膚質檢測儀拍攝,運用多重光譜影像技術對[實施例6]中的同一受試者在使用面膜前與後的面部分析肌膚泛紅面積的數值。肌膚紅色區(深色)是偵測臉部發炎、敏感的狀態,數值越低表示發炎敏感程度越低。Using the VISIA full-face skin quality detector sold by Canfield in the United States to take pictures, use the multiple spectral imaging technology to analyze the value of the skin redness area on the face of the same subject in [Example 6] before and after using the mask. The red area (dark color) of the skin is used to detect the inflammation and sensitivity of the face. The lower the value, the lower the sensitivity of inflammation.
請參閱圖5A,該圖係受試者在使用含本案果香菊萃取物之面膜前與後的瞬效(15分鐘)的肌膚泛紅面積,以受試者使用前的肌膚泛紅面積為100%。其中,受試者使用後的泛紅面積減少至89.4%,使用含本案果香菊萃取物之面膜後,肌膚上的泛紅面積立刻明顯減少,具有良好的肌膚鎮靜功效,避免肌膚發炎敏感。Please refer to Figure 5A, which is the instant (15 minutes) reddened area of the subject's skin before and after using the mask containing the fruity chrysanthemum extract of this case. The reddened area of the skin of the subject before use is 100%. Among them, the reddened area of the subjects after using it was reduced to 89.4%. After using the mask containing the fruity chrysanthemum extract of this case, the reddened area on the skin was significantly reduced immediately, which has a good skin calming effect and prevents skin inflammation and sensitivity.
請參閱圖5B,該圖係受試者在使用含本案果香菊萃取物之面膜前與後的長效(4週)的肌膚泛紅面積,以受試者使用前的肌膚泛紅面積為100%。其中,受試者使用後的泛紅面積減少至91.0%。長期使用含本案果香菊萃取物之面膜後,肌膚上的泛紅面積明顯減少,具有良好的肌膚鎮靜功效,避免肌膚發炎敏感。Please refer to Figure 5B, which is the long-term (4 weeks) skin redness area of the subject before and after using the mask containing the fruity chrysanthemum extract of this case, and the redness area of the skin before the subject's use is 100%. Among them, the reddened area of the subjects after use was reduced to 91.0%. After long-term use of the mask containing the fruity chrysanthemum extract of this case, the reddened area on the skin is significantly reduced, and it has a good skin calming effect, preventing skin inflammation and sensitivity.
[實施例8]:人體實驗-經皮水分散失量的檢測[Example 8]: Human experiment - detection of transdermal water loss
使用購自德國Courage+Khazaka electronic公司之肌膚水分散失檢測探頭TM 300(C+K Multi Probe Adapter System,Germany)對[實施例6]中的同一受試者在使用果香菊萃取物精華液的面膜或安慰劑前與後的面部肌膚進行檢測。該檢測探頭系使用兩端開放的圓柱形腔體在皮膚表面形成相對穩定的測試環境,通過測定不同兩點的水蒸氣壓梯度,計算出經表皮蒸發的水分量,以此來衡量皮膚表面水分流失情況。Use the skin water loss detection probe TM 300 (C+K Multi Probe Adapter System, Germany) purchased from Courage+Khazaka electronic company in Germany to test the same subject in [Example 6] who used the chrysanthemum extract essence. Facial skin was tested before and after the mask or placebo. The detection probe uses a cylindrical cavity with open ends to form a relatively stable test environment on the skin surface. By measuring the water vapor pressure gradient at two different points, the amount of water evaporated through the epidermis is calculated to measure the skin surface moisture. Churn situation.
由圖6可知,相較於使用果香菊萃取物精華液的面膜之前,在持續使用果香菊萃取物精華液的面膜4周後,經表皮水分散失率顯著改善約94.2%。基此可知,果香菊萃取物以外用方式塗敷於肌膚上,確實可使經表皮水分散失率減少,因此提升肌膚的保濕能力,並具改善TEWL值異常之相關皮膚疾病之潛力。It can be seen from Figure 6 that, compared with before using the facial mask of fruity chrysanthemum extract essence, after 4 weeks of continuous use of the facial mask of fruity chrysanthemum extract essence, the water loss rate through the epidermis was significantly improved by about 94.2%. Based on this, it can be seen that the external application of the fruit extract on the skin can indeed reduce the transepidermal water loss rate, thus improving the skin's moisturizing ability, and has the potential to improve skin diseases related to abnormal TEWL values.
綜上所述,任一實施例的果香菊萃取物可用於製備調理肌膚的組合物,該果香菊萃取物具有抗發炎、提高超氧化物歧化酶的活性、強化肌膚屏障、降低肌膚的紅斑生成及/或提升肌膚的保濕度。本發明的果香菊萃取物提高肌膚防禦力,減少氧化壓力形成自由基,並且減少肌膚發炎,避免毛細血管受刺激而產生發炎反應,效鎮靜舒緩肌膚、減少肌膚泛紅區域,以提升肌膚穩定性及避免膚色不均。此外,本發明的果香菊萃取物還強化肌膚屏障,以防止水分散失,使肌膚保持水潤,提供肌膚最佳保護力。In summary, the chrysanthemum extract of any embodiment can be used to prepare a composition for conditioning the skin. The chrysanthemum extract has the functions of anti-inflammation, improving the activity of superoxide dismutase, strengthening the skin barrier, reducing skin Erythema formation and/or increase skin hydration. The fruity chrysanthemum extract of the present invention improves skin defense, reduces oxidative stress to form free radicals, reduces skin inflammation, prevents capillaries from being stimulated to produce inflammatory reactions, calms and soothes skin, reduces skin redness, and improves skin stability. Sexuality and avoid uneven skin tone. In addition, the fruity chrysanthemum extract of the present invention also strengthens the skin barrier to prevent water loss, keeps the skin moisturized, and provides the best protection for the skin.
無。none.
圖1係控制組、對照組及實驗組抑制活性氧物質生成之結果比較圖。###p值<0.001,相較於控制組,***p值<0.001,相較於對照組。 圖2係控制組、對照組及實驗組抵抗紫外線所誘發的白血球介素-8 (IL-8)基因的表現量的結果比較圖。###p值<0.001,相較於控制組,***p值<0.001,相較於對照組。 圖3係受試者使用含有該實驗組的精華液的面膜前與後的肌膚皺紋區域的結果比較圖以及以檢測儀所拍攝的圖像,以使用前受試者的皺紋區域為100%。 圖4A及4B分別係受試者使用含有安慰劑及1%果香菊萃取物的精華液的面膜的肌膚保濕度的瞬效(15分鐘)及長效(4週)的結果比較圖,以使用前受試者的肌膚保濕度為100%。 圖5A及5B分別係受試者使用含有安慰劑及1%果香菊萃取物的精華液的面膜的紅斑生成指數的瞬效(15分鐘)及長效(4週)的結果比較圖,以使用前受試者的紅斑生成指數為100%。 圖6係受試者使用含有安慰劑及1%果香菊萃取物的精華液的面膜的經皮水分散失量的長效(4週)的結果比較圖,以使用前受試者的紅斑生成指數為100%。 Figure 1 is a comparison chart of the results of inhibiting the generation of active oxygen species in the control group, the control group and the experimental group. ### p-value < 0.001 compared to control group, ***p-value < 0.001 compared to control group. Fig. 2 is a comparison chart of the expression level of the interleukin-8 (IL-8) gene induced by the resistance to ultraviolet rays in the control group, the control group and the experimental group. ### p-value < 0.001 compared to control group, ***p-value < 0.001 compared to control group. Figure 3 is a comparison chart of the results of the wrinkled area of the skin before and after the subject uses the mask containing the essence of the experimental group and the image taken by the detector, taking the wrinkled area of the subject before use as 100%. Figures 4A and 4B are the results comparison charts of the instant (15 minutes) and long-term (4 weeks) results of the skin moisturizing degree of the subject using the mask containing the essence of the placebo and 1% fruity chrysanthemum extract, respectively. Before use, the skin moisture level of the subject was 100%. Figures 5A and 5B are the results comparison charts of the instant (15 minutes) and long-term (4 weeks) results of the erythema generation index of the facial mask containing placebo and 1% fruity chrysanthemum extract, respectively. The erythema production index of the subjects before use was 100%. Figure 6 is a long-term (4 weeks) comparison chart of the results of the transdermal water loss of the mask containing the essence of the placebo and 1% fruity chrysanthemum extract, based on the erythema formation of the previous subjects The index is 100%.
Claims (7)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
TW110134713A TWI788019B (en) | 2021-09-16 | 2021-09-16 | Use of chamaemelum nobile extract for manufacturing skincare composition |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
TW110134713A TWI788019B (en) | 2021-09-16 | 2021-09-16 | Use of chamaemelum nobile extract for manufacturing skincare composition |
Publications (2)
Publication Number | Publication Date |
---|---|
TWI788019B true TWI788019B (en) | 2022-12-21 |
TW202313083A TW202313083A (en) | 2023-04-01 |
Family
ID=85795156
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
TW110134713A TWI788019B (en) | 2021-09-16 | 2021-09-16 | Use of chamaemelum nobile extract for manufacturing skincare composition |
Country Status (1)
Country | Link |
---|---|
TW (1) | TWI788019B (en) |
-
2021
- 2021-09-16 TW TW110134713A patent/TWI788019B/en active
Non-Patent Citations (3)
Title |
---|
期刊 Herrera-Carrera et al. Phenolic composition of selectedherbal infusions and their anti-inflammatory effect on a colonic model in vitro in HT-29 cells Cogent Food & Agriculture 1(1) 2015 1059033 * |
期刊 Johnson et al. Safety Assessment of Anthemis nobilis–Derived Ingredients as Used in Cosmetics International Journal of Toxicology 36(s1) 2017 57S-66S; * |
期刊 Srivastava et al. Chamomile: A herbal medicine of the past with bright future Molecular Medicine Report 3(6) 2010 895-901; * |
Also Published As
Publication number | Publication date |
---|---|
TW202313083A (en) | 2023-04-01 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
KR102513930B1 (en) | Anisoxanthos Flavidus Extract for Cosmetic Use | |
US8709511B2 (en) | External preparation composition for skin comprising ginseng flower or ginseng seed extracts | |
JP7275103B2 (en) | Novel cosmetic use of Nephelium lapaceum extract | |
CN108498408B (en) | Whitening essence | |
US20140356468A1 (en) | Composition containing paper mulberry extracts | |
TWI754189B (en) | Use of protea cynaroides extract for enhancing mitochondrial activity of skin cells | |
KR101917645B1 (en) | Cosmetic composition comprising saccharomyces ferment filtrate, asparagus officinalis stem extract and tussilago farfara flower extract | |
JP2024091788A (en) | Use of bixa orellana extract | |
KR20200073313A (en) | Cosmetic composition containing sarcodon aspratus and magnolia liliflora buda fermented extract using Yusung hot spring water | |
KR20140018666A (en) | Composition for skin external application containing fermented soybean extract | |
TW202106323A (en) | Use ofrosmarinus officinaliscallus extract for manufacturing composition inhibiting skin aging and a culture medium for inducingrosmarinus officinaliscallus | |
KR101135264B1 (en) | Cosmetic composition containing the extract of medicinal herb mixture | |
TWI788019B (en) | Use of chamaemelum nobile extract for manufacturing skincare composition | |
US20220054398A1 (en) | Use of rubus fruticosus extract for manufacturing a skincare composition | |
KR20100023372A (en) | Cosmetic composition containing phyllanthus urinaria extrancts | |
JP5855949B2 (en) | Keratin production promoter, hair dye and nail polish | |
KR20120058718A (en) | Composition of skin external application containing Panax ginseng var. Hwangsookjong extracts | |
JP3686394B2 (en) | Anti-aging agent, Maillard reaction inhibitor, collagenase activity inhibitor and cosmetics for preventing skin aging containing these | |
KR20190107454A (en) | Composition Containing Euryale ferox Seed Extract for Anti-Aging of the Skin and Reinforcement of Skin Barrier Function | |
CN115813815A (en) | Use of chamomile juice for preparing a composition for conditioning the skin | |
KR102468538B1 (en) | Skin improvement composition containing wild geranium extract with excellent antioxidant, whitening and anti-inflammatory effects | |
TWI776985B (en) | Use of labisia pumila extract for improving skin elasticity, strength or stability | |
TW202207964A (en) | Use ofrubus fruticosusextract for manufacturing a skincare composition | |
CN114423411A (en) | Use of olive kernel extract in cosmetics or health products | |
JP2022543141A (en) | Novel cosmetic use of willow herb (Epilobium angustifolium) extract |