TW202247837A - Manufacturing method of longan shell extract and use thereof - Google Patents
Manufacturing method of longan shell extract and use thereof Download PDFInfo
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本發明有關一種植物萃取物，特別是指無患子科植物萃取物，由該萃取物所製成的組合物可作為仿曬製劑、頭皮養護產品、洗髮精、口服製劑、食品或生髮水等產品的原料。The present invention relates to a plant extract, in particular to the plant extract of Sapindaceae, and the composition made from the extract can be used as self-tanning preparation, scalp care product, shampoo, oral preparation, food or hair tonic and other products raw materials.
無患子科植物包含：荔枝、瓜拿納、龍眼、紅毛丹、蜜果、阿開木、與無患子，其富含營養，適用於作為美容品、保養品、護膚品、或護髮用品等產品的原料，具有高經濟價值。因此，將無患子科植物部位進行萃取，並將其有效成分用於製備可施用於人體或供人體食用之產品實為時勢所趨。Sapindaceae plants include: lychee, guarana, longan, rambutan, honey fruit, acai wood, and sapindus, which are rich in nutrition and are suitable for beauty products, skin care products, skin care products, or hair care products. Raw materials with high economic value. Therefore, it is the trend of the times to extract the parts of Sapindaceae plants and use their active ingredients to prepare products that can be applied to the human body or eaten by the human body.
本發明之目的在於利用無患子科植物部位所含之有效成分，並將其製成可供人體使用或食用之產品。The purpose of the present invention is to utilize the active ingredients contained in the parts of Sapindaceae plants and make them into products that can be used or eaten by the human body.
為解決上述之問題，本發明提供一種無患子科植物萃取物之用途，係用於製備促進黑色素生成或強化髮根的組合物。In order to solve the above problems, the present invention provides a use of a Sapindaceae plant extract, which is used to prepare a composition for promoting melanin production or strengthening hair roots.
更佳者，其中該無患子科植物萃取物之製備方法包含： 一浸泡步驟，提取一重量比例為1：1至1：12的無患子科植物部位以及水，並以水將該無患子科植物部位覆蓋浸泡，以製成一無患子科植物部位懸浮液；以及 一萃取步驟，於溫度條件10°C至90°C下，以總能量100至500 w的超音波能量連續快速振盪萃取該無患子科植物部位懸浮液。More preferably, the preparation method of the Sapindaceae plant extract comprises: a soaking step, extracting a Sapindaceae plant part and water in a weight ratio of 1:1 to 1:12, and covering the Sapindaceae plant part with water Soaking to prepare a suspension of a Sapindaceae plant part; and an extraction step of extracting the Sapindaceae plant part with continuous and rapid oscillation of ultrasonic energy with a total energy of 100 to 500 W at a temperature of 10°C to 90°C suspension.
更佳者，其中該無患子科植物萃取物之製備方法更包含：一研磨步驟，係於進行該浸泡步驟前，先將該無患子科植物部位磨碎為粉末、碎塊、或小顆粒。More preferably, the preparation method of the Sapindaceae plant extract further comprises: a grinding step, which is to grind the Sapindaceae plant part into powder, fragments, or small particles before the soaking step.
更佳者，其中該無患子科植物萃取物之製備方法更包含： 一冷凍步驟，係於進行該研磨步驟前，先將該無患子科植物部位冷凍。More preferably, the preparation method of the Sapindaceae plant extract further comprises: a freezing step, which is to freeze the parts of the Sapindaceae plant before the grinding step.
更佳者，其中該無患子科植物萃取物之製備方法更包含： 一離心步驟，對該快速振盪萃取後的無患子科植物部位懸浮液進行高速離心，以去除底部沉澱物並收集上清液。More preferably, the preparation method of the Sapindaceae plant extract further comprises: a centrifugation step, performing high-speed centrifugation on the rapidly shaken extracted Sapindaceae plant part suspension to remove the bottom sediment and collect the supernatant.
更佳者，其中該無患子科植物萃取物之製備方法更包含：一過濾步驟，將該上清液以真空篩檢器過濾，以獲得一無患子科植物部位萃取液。More preferably, the preparation method of the Sapindaceae plant extract further comprises: a filtering step, filtering the supernatant with a vacuum filter to obtain a Sapindaceae plant part extract.
更佳者，其中該無患子科植物萃取物之製備方法更包含：一乾燥步驟，利用減壓濃縮機及真空冷凍乾燥機將該無患子科植物部位萃取液凍乾，以取得一無患子科植物部位乾燥萃取物。More preferably, the preparation method of the Sapindaceae plant extract further comprises: a drying step, using a decompression concentrator and a vacuum freeze dryer to freeze-dry the Sapindaceae plant part extract to obtain a dried Sapindaceae plant part Extracts.
更佳者，其中該無患子科植物萃取物之使用劑量為：於每平方公分面積之頭皮或皮膚上施用0.05至0.15 mg。More preferably, the dosage of the Sapindaceae plant extract is: 0.05 to 0.15 mg per square centimeter of scalp or skin.
更佳者，其中：該無患子科植物部位係指一無患子科植物之全株、根、莖、葉、花、果肉、果皮、種子、或果殼；以及 該無患子科植物為荔枝、瓜拿納、龍眼、紅毛丹、蜜果、阿開木、或無患子。More preferably, wherein: the part of the Sapindaceae plant refers to the whole plant, root, stem, leaf, flower, pulp, peel, seed, or husk of a Sapindaceae plant; and the Sapindaceae plant is litchi, guarana , longan, rambutan, honey fruit, Akai wood, or Sapindus.
更佳者，其中： 該無患子科植物部位係指一無患子科植物之果殼；以及 該無患子科植物為龍眼。綜上所述，本發明所得之無患子科植物萃取物可施用於人體皮膚或毛髮，具有促黑之功效，由於其為天然物所得，因此為對身體無害的促黑產品。除了促黑之功效外，該無患子科植物萃取物或添加有該無患子科植物萃取物所製成之產品亦可修復頭皮、保養頭皮、恢復頭皮皮膚光澤度、強化髮根、或促進毛髮生長。此外，該無患子科植物萃取物更可作為仿曬製劑、頭皮養護產品、洗髮精、口服製劑、食品或生髮水等產品的原料。More preferably, wherein: the part of the Sapindaceae plant refers to the husk of a Sapindaceae plant; and the Sapindaceae plant is longan. To sum up, the Sapindaceae plant extract obtained in the present invention can be applied to human skin or hair, and has the effect of promoting darkening. Since it is obtained from natural products, it is a harmless darkening product. In addition to the effect of promoting darkening, the Sapindaceae plant extract or products made with the Sapindaceae plant extract can also repair the scalp, maintain the scalp, restore the luster of the scalp skin, strengthen hair roots, or promote hair growth. In addition, the Sapindaceae plant extract can be used as a raw material for products such as self-tanning preparations, scalp care products, shampoos, oral preparations, food or hair tonic.
為讓本發明上述及/或其他目的、功效、特徵更明顯易懂，下文特舉較佳實施方式，作詳細說明於下：In order to make the above and/or other purposes, effects, and features of the present invention more obvious and understandable, the preferred implementation modes are specifically cited below, which are described in detail below:
本發明提供一種無患子科植物萃取物之用途，係用於製備促進黑色素生成或強化髮根的組合物。於一較佳實施例中，由該萃取物所製成的組合物可作為仿曬製劑、頭皮養護產品、洗髮精、口服製劑、食品或生髮水等產品的原料。於另一較佳實施例中，將該組合物或該組合物所製成之產品施用於皮膚或毛髮時，可促進黑色素之生成。於又一較佳實施例中，該組合物或該組合物所製成之產品除達到頭髮變黑之功效，亦可修復頭皮、保養頭皮、恢復頭皮皮膚光澤度、強化髮根、或促進毛髮生長，且不以此為限。於又一較佳實施例中，其中該組合物包含： 一無患子科植物萃取物；以及 一添加成分，其中該添加成分係選自以下所組成之群組：水、酒精、中藥萃取物、植物萃取物、香氛、精油、礦物質、微量元素、或以上之結合。於又一較佳實施例中，該添加成分包含：水、酒精、何首烏萃取液、褐藻萃取液、土肉桂萃取液、微量元素、PGA、雪松精油、百里香精油、香精、或以上之結合。於又一較佳實施例中，何首烏萃取液具有促進毛髮生長、維護頭髮健康色澤、保濕等功效；褐藻萃取液、與土肉桂萃取液具有抗氧化之特性；PGA為護髮素，可滋養、保護頭髮；雪松精油與百里香精油能促進皮膚的潔淨與健康、使頭皮放鬆、與舒緩頭皮。於又一較佳實施例中，頭皮養護產品可以軟膏、乳霜劑、皮膚外用注射劑或噴霧之形式給予個體的頭皮。於又一較佳實施例中，當製劑為噴霧時，其載體為乳糖、滑石、矽石、氫氧化鋁、矽酸鈣或聚酰胺粉末，推進劑為氯氟烃、丙烷/丁烷或二甲醚。於又一較佳實施例中，為延長無患子科植物萃取物對頭皮之功效，頭皮養護產品可以氣溶膠之形式給與個體的頭皮，其包含氣溶膠儲備原液（無患子科植物萃取液）、氣溶膠推進劑（至少一選自丙烷、丁烷、n-丁烷、異丁烷、戊烷、n-戊烷、異戊烷、新戊烷氣體、液化石油氣、液化天然氣、二甲醚）。於又一較佳實施例中，洗髮精之基質可以是溶液、水包油乳劑、懸浮液、微乳液或皮膚洗劑。當基質為溶液時，溶劑可以是水、乙醇、異丙醇、碳酸乙酯、乙酸乙酯、芐醇、苯甲酸芐酯、丙二醇、1,3-丁二醇、甘油脂肪酸酯、聚乙二醇或山梨醇酐的脂肪酸酯。於又一較佳實施例中，口服製劑包含固體製劑或液體製劑。其中：固體製劑可以是錠劑、丸劑、粉末、顆粒劑、軟或硬膠囊，內含無患子科植物萃取物與至少一賦形劑，如澱粉、碳酸鈣、蔗糖、乳糖、明膠。液體製劑可以是懸浮液、內用液體藥物、乳劑、糖漿，內含無患子科植物萃取物、稀釋劑如水或液體石蠟、賦形劑如保濕劑、甜味劑、芳族、防腐劑等。於又一較佳實施例中，食品包含食物、保健食品或膳食補充品。可以理解地，食物可以是口香糖、焦糖、糖果、冰棒等；保健食品可以是保健飲料，內含無患子科植物萃取物、水、水果果汁；膳食補充品可以是維他命和礦物質，內含無患子科植物萃取物、各營養素、維他命、調味劑等，但不以此為限。於又一較佳實施例中，仿曬製劑可與一載體共同組成製劑，載體包括賦型劑或添加劑，共同形成液狀、乳液狀、乳霜狀或膠狀，可施予個體的皮膚。於又一較佳實施例中，可使用無患子科植物萃取物所製成的組合物來作為仿曬製劑、頭皮養護產品、洗髮精、口服製劑、食品或生髮水等產品的原料。The invention provides the use of a Sapindaceae plant extract, which is used for preparing a composition for promoting melanin production or strengthening hair roots. In a preferred embodiment, the composition made from the extract can be used as a raw material for products such as self-tanning preparations, scalp care products, shampoos, oral preparations, food or hair tonic. In another preferred embodiment, when the composition or the product made of the composition is applied to the skin or hair, the production of melanin can be promoted. In yet another preferred embodiment, the composition or the product made of the composition can not only achieve the effect of hair blackening, but also repair the scalp, maintain the scalp, restore the gloss of the scalp skin, strengthen the hair root, or promote hair growth. grow, but not limited to. In yet another preferred embodiment, wherein the composition comprises: a Sapindaceae plant extract; and an additional ingredient, wherein the additional ingredient is selected from the group consisting of: water, alcohol, traditional Chinese medicine extract, plant Extracts, fragrances, essential oils, minerals, trace elements, or a combination of the above. In yet another preferred embodiment, the added ingredients include: water, alcohol, Polygonum multiflorum extract, brown algae extract, soil cinnamon extract, trace elements, PGA, cedarwood essential oil, thyme essential oil, essence, or a combination of the above. In yet another preferred embodiment, the extract of Polygonum multiflorum has the functions of promoting hair growth, maintaining the healthy color of hair, and moisturizing; the extract of brown algae and the extract of cinnamon have anti-oxidation properties; Protects hair; cedarwood essential oil and thyme essential oil promote clean and healthy skin, relax the scalp, and soothe the scalp. In yet another preferred embodiment, the scalp care product can be administered to the individual's scalp in the form of ointment, cream, external injection or spray. In yet another preferred embodiment, when the formulation is a spray, the carrier is lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder, and the propellant is chlorofluorocarbon, propane/butane or bis methyl ether. In yet another preferred embodiment, in order to prolong the effect of Sapindaceae plant extracts on the scalp, scalp maintenance products can be given to the scalp of individuals in the form of aerosols, which include aerosol stock solution (Sapindaceae plant extracts), air Sol propellant (at least one selected from propane, butane, n-butane, isobutane, pentane, n-pentane, isopentane, neopentane gas, liquefied petroleum gas, liquefied natural gas, dimethyl ether) . In yet another preferred embodiment, the base of the shampoo can be a solution, an oil-in-water emulsion, a suspension, a microemulsion or a skin lotion. When the matrix is a solution, the solvent can be water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butanediol, glycerol fatty acid esters, polyethylene glycol Fatty acid esters of diols or sorbitan. In yet another preferred embodiment, the oral formulation comprises solid formulation or liquid formulation. Wherein: the solid preparation can be lozenge, pill, powder, granule, soft or hard capsule, containing Sapindaceae plant extract and at least one excipient, such as starch, calcium carbonate, sucrose, lactose, gelatin. Liquid preparations can be suspensions, liquid medicines for internal use, emulsions, syrups, containing Sapindaceae plant extracts, diluents such as water or liquid paraffin, excipients such as humectants, sweeteners, aromatics, preservatives, etc. In yet another preferred embodiment, the food includes food, health food or dietary supplement. Understandably, the food can be chewing gum, caramel, candy, popsicles, etc.; the health food can be a health drink containing Sapindaceae plant extracts, water, fruit juice; the dietary supplement can be vitamins and minerals containing Sapindus family plant extracts, various nutrients, vitamins, flavoring agents, etc., but not limited thereto. In yet another preferred embodiment, the self-tanning formulation can be combined with a carrier, which includes excipients or additives, to form a liquid, emulsion, cream or jelly, which can be applied to the individual's skin. In yet another preferred embodiment, the composition made from the extract of Sapindaceae plants can be used as raw materials for products such as self-tanning preparations, scalp care products, shampoos, oral preparations, food or hair tonic.
更佳者，其中該無患子科植物萃取物之製備方法包含： 一浸泡步驟，以水將一無患子科植物部位覆蓋浸泡，以製成一無患子科植物部位懸浮液；以及 一萃取步驟，於溫度條件10°C至90°C下，以總能量100至500 w的超音波能量連續快速振盪萃取該無患子科植物部位懸浮液。於一較佳實施例中，該超音波能量為200至400 w；較佳者，該超音波能量為300 w。於一較佳實施例中，以超音波能量對該無患子科植物部位懸浮液進行萃取時，係以震盪3分鐘再休息2分鐘為一循環，持續進行6至10個循環；較佳者為7至9個循環；更佳者為8個循環。於又一較佳實施例中，該溫度條件為20°C至80°C；較佳者，該溫度條件為30°C至70°C；較佳者，該溫度條件為40°C至60°C ；較佳者，該溫度條件為50°C至60°C 。於又一較佳實施例中，可重複進行該萃取步驟，直到取得最大活性之無患子科植物萃取物。於又一較佳實施例中，除了可利用超音波能量對該無患子科植物部位懸浮液進行萃取，亦可透過二氧化碳超臨界流體萃取、熱水萃取、加熱萃取、冷沉澱萃取、回流萃取、冷凝回流萃取、發酵、或酶處理之方式來對該無患子科植物部位懸浮液進行萃取，且不以此為限。於另一較佳實施例中，該無患子科植物部位可浸泡於一溶劑以進行萃取，其中該溶劑係選自以下所組成之群組：水、乙醇、丙酮、乙酸乙脂、氯化納水溶液、氯化鉀水溶液、氯化鈣水溶液、氯化鎂水溶液或氯化鈉乙醇溶液、氯化鉀乙醇溶液、氯化鈣乙醇溶液或氯化鎂乙醇溶液、有機溶劑、***、甲基乙基酮、氯仿、其他有機溶劑（C1-C6低醇類、丁二醇、丙二醇、己二醇、二丙二醇）、或以上之結合。於又一較佳實施例中，該溶液與該無患子科植物部位之重量比為5：1。More preferably, the preparation method of the Sapindaceae plant extract comprises: a soaking step, covering and soaking a Sapindaceae plant part with water to prepare a Sapindaceae plant part suspension; and an extraction step, under temperature conditions At 10°C to 90°C, the suspension of Sapindaceae plant parts is extracted with continuous and rapid oscillation of ultrasonic energy with a total energy of 100 to 500 W. In a preferred embodiment, the ultrasonic energy is 200 to 400 w; preferably, the ultrasonic energy is 300 w. In a preferred embodiment, when the suspension of the Sapindaceae plant part is extracted with ultrasonic energy, a cycle of shaking for 3 minutes and resting for 2 minutes is used as a cycle, and the cycle is continued for 6 to 10 cycles; preferably 7 Up to 9 cycles; more preferably 8 cycles. In yet another preferred embodiment, the temperature condition is 20°C to 80°C; preferably, the temperature condition is 30°C to 70°C; preferably, the temperature condition is 40°C to 60°C °C; preferably, the temperature condition is 50°C to 60°C. In yet another preferred embodiment, the extraction step can be repeated until the maximum activity of the Sapindaceae plant extract is obtained. In yet another preferred embodiment, in addition to using ultrasonic energy to extract the suspension of the Sapindaceae plant parts, it can also be extracted through carbon dioxide supercritical fluid extraction, hot water extraction, heating extraction, cryoprecipitation extraction, reflux extraction, condensation The method of reflux extraction, fermentation, or enzyme treatment is used to extract the suspension of the Sapindaceae plant parts, but not limited thereto. In another preferred embodiment, the plant parts of Sapindaceae can be soaked in a solvent for extraction, wherein the solvent is selected from the group consisting of water, ethanol, acetone, ethyl acetate, sodium chloride aqueous solution , potassium chloride aqueous solution, calcium chloride aqueous solution, magnesium chloride aqueous solution or sodium chloride ethanol solution, potassium chloride ethanol solution, calcium chloride ethanol solution or magnesium chloride ethanol solution, organic solvents, ether, methyl ethyl ketone, chloroform, others Organic solvents (C1-C6 low alcohols, butylene glycol, propylene glycol, hexylene glycol, dipropylene glycol), or a combination of the above. In yet another preferred embodiment, the weight ratio of the solution to the Sapindaceae plant part is 5:1.
更佳者，在浸泡步驟中，於以水將無患子科植物部位覆蓋浸泡時，無患子科植物部位與水之重量比例為1：1至1：12。於一較佳實施例中，該無患子科植物部位與水之重量比例為1：2、1：3、1：4、1：5、1：6、1：7、1：8、1：9或1：11；較佳者，該無患子科植物部位與水之重量比例為1：10。於另一較佳實施例中，可將無患子科植物部位放入溶液後馬上進行萃取步驟，但不以此為限，實際之浸泡時間可依各種狀況進行調整。於又一較佳實施例中，其中該無患子科植物萃取物之製備方法更包含：一研磨步驟，係於進行該浸泡步驟前，先將該無患子科植物部位磨碎為粉末、碎塊、或小顆粒。 於又一較佳實施例中，其中該無患子科植物萃取物之製備方法更包含： 一冷凍步驟，係於進行該研磨步驟前，先將該無患子科植物部位冷凍。於一較佳實施例中，除了可以冷凍之方式乾燥、固化該無患子科植物部位，亦可透過烘烤、曝曬、蒸發、或噴霧之方式進行乾燥，但不以此為限。於又一較佳實施例中，其中該無患子科植物萃取物之製備方法更包含： 一離心步驟，對該快速振盪萃取後的無患子科植物部位懸浮液進行高速離心，以去除底部沉澱物並收集上清液。於又一較佳實施例中，其中該無患子科植物萃取物之製備方法更包含：一過濾步驟，將該上清液以真空篩檢器過濾，以獲得一無患子科植物部位萃取液。於又一較佳實施例中，其中該無患子科植物萃取物之製備方法更包含：一乾燥步驟，利用減壓濃縮機及真空冷凍乾燥機將該無患子科植物部位萃取液凍乾，以取得一無患子科植物部位乾燥萃取物。 於又一較佳實施例中，進行真空過濾之溫度條件為45至70℃，但不以此為限。於又一較佳實施例中，該篩檢器為一400目篩網，但不以此為限。於又一較佳實施例中，該無患子科植物部位萃取液之濃度較佳為50至300 μg/mL；較佳者為250 μg/mL。More preferably, in the soaking step, when covering and soaking the parts of the Sapindaceae plant with water, the weight ratio of the plant part of the Sapindaceae plant to the water is 1:1 to 1:12. In a preferred embodiment, the weight ratio of the Sapindaceae plant parts to water is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 Or 1:11; more preferably, the weight ratio of the Sapindaceae plant parts to water is 1:10. In another preferred embodiment, the extracting step can be carried out immediately after putting the plant part of Sapindaceae into the solution, but it is not limited thereto, and the actual soaking time can be adjusted according to various situations. In yet another preferred embodiment, the preparation method of the Sapindaceae plant extract further comprises: a grinding step, which is to grind the Sapindaceae plant part into powder, pieces, or small particles. In yet another preferred embodiment, the preparation method of the Sapindaceae plant extract further comprises: a freezing step, which is to freeze the parts of the Sapindaceae plant before the grinding step. In a preferred embodiment, in addition to drying and solidifying the parts of the Sapindaceae plant by freezing, it can also be dried by roasting, exposing to the sun, evaporating, or spraying, but it is not limited thereto. In yet another preferred embodiment, the preparation method of the Sapindaceae plant extract further comprises: a centrifugation step, performing high-speed centrifugation on the rapidly shaken extracted Sapindaceae plant part suspension to remove the bottom sediment and collect supernatant. In yet another preferred embodiment, the preparation method of the Sapindaceae plant extract further comprises: a filtering step, filtering the supernatant with a vacuum filter to obtain a Sapindaceae plant part extract. In yet another preferred embodiment, the preparation method of the Sapindaceae plant extract further comprises: a drying step, using a decompression concentrator and a vacuum freeze dryer to freeze-dry the Sapindaceae plant part extract to obtain a Dry extract of plant parts of Sapinaceae. In yet another preferred embodiment, the temperature condition for vacuum filtration is 45 to 70° C., but not limited thereto. In yet another preferred embodiment, the screening device is a 400-mesh screen, but not limited thereto. In yet another preferred embodiment, the concentration of the extract from parts of Sapindaceae plants is preferably 50 to 300 μg/mL; more preferably 250 μg/mL.
更佳者，該無患子科植物萃取物係自一無患子科植物之根、莖、葉、花、果實、種子、或果殼中提取。於一較佳實施例中，該無患子科植物部位係指一無患子科植物之全株、根、莖、葉、花、果肉、果皮、種子、或果殼，但不以此為限。於另一較佳實施例中，該無患子科植物為荔枝、瓜拿納、龍眼、紅毛丹、蜜果、阿開木、或無患子，但不以此為限。於又一較佳實施例中，其中該無患子科植物萃取物之使用劑量為：於每平方公分面積之頭皮或皮膚上施用0.05至0.15 mg。More preferably, the Sapindaceae plant extract is extracted from the root, stem, leaf, flower, fruit, seed, or husk of a Sapindaceae plant. In a preferred embodiment, the part of the Sapindaceae plant refers to the whole plant, root, stem, leaf, flower, pulp, peel, seed, or husk of a Sapindaceae plant, but it is not limited thereto. In another preferred embodiment, the Sapindaceae plant is litchi, guarana, longan, rambutan, honey fruit, acai, or Sapindus, but not limited thereto. In yet another preferred embodiment, the dosage of the Sapindaceae plant extract is: 0.05 to 0.15 mg per square centimeter of scalp or skin.
更佳者，該無患子科植物萃取物係指一龍眼殼萃取物。於一較佳實施例中，該龍眼殼萃取物之製備方法包含：一浸泡步驟，以水將龍眼殼覆蓋浸泡，以製成一龍眼殼懸浮液；以及 一萃取步驟，於溫度條件10°C至90°C下，以總能量100至500 w的超音波能量連續快速振盪萃取該龍眼殼懸浮液。於另一較佳實施例中，在浸泡步驟中，於以水將龍眼殼覆蓋浸泡時，龍眼殼與水之重量比例為1：1至1：12。 於又一較佳實施例中，其中該龍眼殼萃取物之製備方法更包含：一研磨步驟，係於進行該浸泡步驟前，先將該龍眼殼磨碎為粉末、碎塊、或小顆粒。於又一較佳實施例中，其中該龍眼殼萃取物之製備方法更包含： 一冷凍步驟，係於進行該研磨步驟前，先將該龍眼殼冷凍。於又一較佳實施例中，其中該龍眼殼萃取物之製備方法更包含： 一離心步驟，對該快速振盪萃取後的龍眼殼懸浮液進行高速離心，以去除底部沉澱物並收集上清液。於又一較佳實施例中，其中該龍眼殼萃取物之製備方法更包含：一過濾步驟，將該上清液以真空篩檢器過濾，以獲得一龍眼殼萃取液。於又一較佳實施例中，其中該龍眼殼萃取物之製備方法更包含：一乾燥步驟，利用減壓濃縮機及真空冷凍乾燥機將該龍眼殼萃取液凍乾，以取得一龍眼殼乾燥萃取物。於又一較佳實施例中，其中將該組合物施用於皮膚或毛髮時，可促進黑色素之生成。於又一較佳實施例中，其中該龍眼殼萃取物之濃度為50至300 µg/mL。於又一較佳實施例中，其中該龍眼殼萃取物之使用劑量為：於每平方公分面積之頭皮或皮膚上施用0.05至0.15 mg。More preferably, the Sapindaceae plant extract refers to a longan shell extract. In a preferred embodiment, the preparation method of the longan shell extract comprises: a soaking step, soaking the longan shell with water to make a longan shell suspension; and an extraction step, at a temperature of 10°C At 90°C, the longan shell suspension was extracted with continuous and rapid oscillation of ultrasonic energy with a total energy of 100 to 500 w. In another preferred embodiment, in the soaking step, when the longan shell is covered and soaked with water, the weight ratio of the longan shell to water is 1:1 to 1:12. In yet another preferred embodiment, the preparation method of the longan shell extract further comprises: a grinding step, which is to grind the longan shell into powder, pieces, or small particles before the soaking step. In yet another preferred embodiment, the preparation method of the longan shell extract further comprises: a freezing step, which is to freeze the longan shell before the grinding step. In yet another preferred embodiment, the preparation method of the longan shell extract further comprises: a centrifugation step, performing high-speed centrifugation on the longan shell suspension after the rapid shaking and extraction, to remove the bottom sediment and collect the supernatant . In yet another preferred embodiment, the preparation method of the longan shell extract further comprises: a filtering step, filtering the supernatant with a vacuum filter to obtain a longan shell extract. In yet another preferred embodiment, the preparation method of the longan shell extract further includes: a drying step, using a vacuum concentrator and a vacuum freeze dryer to freeze-dry the longan shell extract to obtain a dried longan shell Extracts. In yet another preferred embodiment, when the composition is applied to the skin or hair, it can promote the production of melanin. In yet another preferred embodiment, the concentration of the longan shell extract is 50 to 300 µg/mL. In yet another preferred embodiment, the dosage of the longan shell extract is: 0.05 to 0.15 mg per square centimeter of scalp or skin.
以下提供龍眼殼萃取物之相關試驗，以作證本發明之功效。The relevant tests of the longan shell extract are provided below to prove the efficacy of the present invention.
實施例1：龍眼殼萃取物的製備方法。Embodiment 1: the preparation method of longan shell extract.
首先，將乾燥的龍眼殼研磨後，利用破碎機進行破碎處理。接著，將滅菌之去離子水與破碎之龍眼殼以10：1之比例混合後，利用超音波萃取機獲得龍眼殼萃取液。其參數設定為溫度50~60°C，功率300瓦，震3分鐘，休息2分鐘，以此循環進行40分鐘。萃取液再經由真空過濾法，即可獲得純淨的龍眼殼萃取液。First, the dried longan shells are ground and crushed with a crusher. Next, after mixing the sterilized deionized water and the broken longan shell at a ratio of 10:1, the longan shell extract is obtained by using an ultrasonic extraction machine. Its parameters are set as temperature 50-60°C, power 300 watts, vibration for 3 minutes, rest for 2 minutes, and this cycle is carried out for 40 minutes. The extract is then subjected to vacuum filtration to obtain pure longan shell extract.
實施例2：龍眼殼萃取物於酪胺酸酶之活性試驗。Example 2: Activity test of longan shell extract on tyrosinase.
酪胺酸酶為黑色素生成之一關鍵酵素，故本發明以酪胺酸酶活性來評估龍眼殼萃取液是否具促黑之功效。為配置不同濃度之龍眼殼萃取液，取由實施例1所得之龍眼殼萃取液作進一步濃縮、冷凍乾燥，以獲得龍眼殼乾燥萃取物。該萃取物再以純水定量回溶，配置成50、100及250 μg/mL之龍眼殼萃取液。首先，將 20 μL龍眼殼萃取液及25 μL酪胺酸酶（25 unit）加入96孔盤中，混合均勻後於室溫下靜置10分鐘。接著，加入155 μL L-tyrosine（2.5 mM）或L-Dopa，混合後反應1小時，再利用微盤分析儀（ELISA reader）讀取波長450奈米的吸光值。酪胺酸酶活性抑制率是由以下公式計算：[(A-B)-(C-D)]/(A-B)×100。其中，A為未加龍眼殼萃取液但加入酪胺酸酶的吸光值；B為未加龍眼殼萃取液及酪胺酸酶的吸光值；C為加入龍眼殼萃取液及酪胺酸酶的吸光值；D為加入龍眼殼萃取液但未加酪胺酸酶的吸光值。前述酪胺酸酶活性試驗之結果如圖1所示。由圖1之結果可知，50、100、250 µg/mL龍眼殼萃取液對於酪胺酸酶抑制率呈現負值，分別為-1.2%、-18.8%和-31.3%，顯示龍眼殼萃取液可提高酪胺酸酶之活性。其中，維生素C（ascorbic acid, AA）為正向標準品。Tyrosinase is one of the key enzymes in the production of melanin, so the present invention uses tyrosinase activity to evaluate whether the longan shell extract has the effect of promoting melanin. In order to prepare longan shell extracts with different concentrations, the longan shell extract obtained in Example 1 was further concentrated and freeze-dried to obtain dry longan shell extracts. The extract was quantitatively redissolved in pure water to prepare 50, 100 and 250 μg/mL longan shell extracts. First, add 20 μL longan shell extract and 25 μL tyrosinase (25 unit) into a 96-well plate, mix well and let stand at room temperature for 10 minutes. Next, add 155 μL of L-tyrosine (2.5 mM) or L-Dopa, mix and react for 1 hour, and then use a microdisk analyzer (ELISA reader) to read the absorbance at a wavelength of 450 nm. The inhibition rate of tyrosinase activity was calculated by the following formula: [(A-B)-(C-D)]/(A-B)×100. Among them, A is the absorbance value without adding longan shell extract but adding tyrosinase; B is the absorbance value without adding longan shell extract and tyrosinase; C is adding longan shell extract and tyrosinase Absorbance; D is the absorbance of adding longan shell extract but not adding tyrosinase. The results of the aforementioned tyrosinase activity test are shown in FIG. 1 . It can be seen from the results in Figure 1 that the 50, 100, and 250 µg/mL longan shell extracts showed negative values for the inhibition rate of tyrosinase, which were -1.2%, -18.8% and -31.3%, respectively, indicating that the longan shell extracts could Improve the activity of tyrosinase. Among them, vitamin C (ascorbic acid, AA) is the positive standard.
實施例3 ：龍眼殼萃取物於細胞外之黑色素含量測定。Embodiment 3: Determination of extracellular melanin content of longan shell extract.
於6孔培養盤中接種黑色素瘤細胞（B16 cells），並以含有α-MSH（100 nM）、龍眼殼萃取液、維他命C（AA）或熊果素（arbutin）之培養液培養72小時。接著，取100 µL培養液至96孔盤中，再利用微盤分析儀（ELISA reader）讀取波長405奈米的吸光值。細胞外黑色素含量是由以下公式計算：B/A×100。其中，A為控制組的吸光值；B為各組別的吸光值。前述細胞外黑色素含量測定之結果如圖2所示。由圖2之結果可知，細胞經龍眼殼萃取液（50、100、250 µg/mL）作用後，細胞外黑色素含量分別為112.3%、120.1%和121.8%，顯示龍眼殼萃取液具促進細胞外黑色素生成之功效，且效能＞10%。其中，α-MSH為正向對照組，其細胞外黑色素含量為119.2%；維他命C (AA)和熊果素(arbutin)為負向對照組。Melanoma cells (B16 cells) were inoculated in a 6-well culture dish and cultured for 72 hours in a medium containing α-MSH (100 nM), longan shell extract, vitamin C (AA) or arbutin. Next, take 100 µL of the culture solution into a 96-well plate, and then use a microplate analyzer (ELISA reader) to read the absorbance at a wavelength of 405 nm. The extracellular melanin content was calculated by the following formula: B/A×100. Among them, A is the absorbance value of the control group; B is the absorbance value of each group. The results of the measurement of the aforementioned extracellular melanin content are shown in FIG. 2 . It can be seen from the results in Figure 2 that after the cells were treated with longan shell extract (50, 100, 250 µg/mL), the extracellular melanin content was 112.3%, 120.1% and 121.8%, respectively, indicating that the longan shell extract has the effect of promoting extracellular melanin. The effect of melanin production, and the efficacy> 10%. Among them, α-MSH was the positive control group, and its extracellular melanin content was 119.2%; vitamin C (AA) and arbutin (arbutin) were the negative control groups.
實施例4 ：龍眼殼萃取物於細胞內酪胺酸酶活性試驗。Embodiment 4: Longan shell extract in intracellular tyrosinase activity test.
酪胺酸酶為黑色素生成之一關鍵酵素，故本發明以酪胺酸酶活性來評估龍眼殼萃取液是否具促黑之功效。於6孔培養盤中接種黑色素瘤細胞（B16 cells），並以含有α-MSH（100 nM）、龍眼殼萃取液、維他命C（AA）或熊果素（arbutin）之培養液培養72小時。接著，移除培養液、以PBS清洗後，加入0.5% Triton-100反應5分鐘。最後，加入10 mM L-DOPA，於37˚C、避光之狀況下反應，再以酵素免疫分析儀(BioTek, SynergyTM2, USA)讀取波長475奈米的吸光值。細胞內酪胺酸酶活性是由以下公式計算：B/A×100。其中，A為控制組的吸光值；B為各組別的吸光值。前述細胞內酪胺酸酶活性試驗之結果如圖3所示。由圖3之結果可知，細胞經龍眼殼萃取液（50、100、250 µg/mL）作用後，細胞內酪胺酸酶活性分別為119.4%、120.7%和122.1%，顯示龍眼殼萃取液具促進細胞內酪胺酸酶活性之功效，且效能＞10%。其中，α-MSH為正向對照組，其細胞內酪胺酸酶活性為118.6%；維他命C（AA）和熊果素（arbutin）為負向對照組。Tyrosinase is one of the key enzymes in the production of melanin, so the present invention uses tyrosinase activity to evaluate whether the longan shell extract has the effect of promoting melanin. Melanoma cells (B16 cells) were inoculated in a 6-well culture dish and cultured for 72 hours in a medium containing α-MSH (100 nM), longan shell extract, vitamin C (AA) or arbutin. Next, the culture medium was removed, washed with PBS, and 0.5% Triton-100 was added to react for 5 minutes. Finally, 10 mM L-DOPA was added and reacted at 37°C in the dark, and the absorbance at a wavelength of 475 nm was read with an enzyme immunoassay analyzer (BioTek, SynergyTM2, USA). Intracellular tyrosinase activity was calculated by the following formula: B/A×100. Among them, A is the absorbance value of the control group; B is the absorbance value of each group. The results of the aforementioned intracellular tyrosinase activity test are shown in FIG. 3 . It can be seen from the results in Figure 3 that after the cells were treated with longan shell extract (50, 100, 250 µg/mL), the intracellular tyrosinase activities were 119.4%, 120.7% and 122.1%, respectively, showing that the longan shell extract has The effect of promoting intracellular tyrosinase activity, and the effect is >10%. Among them, α-MSH was the positive control group, and its intracellular tyrosinase activity was 118.6%; vitamin C (AA) and arbutin (arbutin) were the negative control groups.
實施例5： 龍眼殼萃取物於黑色素生成相關基因表現之測定。Example 5: Determination of the expression of genes related to melanin production by longan shell extract.
已知黑色素生成受到MC1R、MITF、Tyrosinase、TRP-1與TRP-2等基因之調節，因此進一步檢測龍眼殼萃取液對黑色素生成相關基因表現量的影響。首先，將黑色素瘤細胞（B16 cells）以1×10 5/mL之密度接種於6孔盤中。培養24小時後，以含有250 µg/mL龍眼殼萃取液之培養液作用72小時。收集細胞後，使用RNA萃取試劑（購自Protech公司，台灣，Cat No. PT-KP200CT）抽取上述細胞之RNA，再利用MMLV反轉錄酶將RNA反轉錄為cDNA，接著使用Quantitect SYBR Green RT-PCR套組（購自Qiagen公司，德國）以螢光偵測系統（prism 7700定序偵測系統）進行定量即時反轉錄聚合酶連鎖反應，條件為50°C反應30分鐘，95°C反應15分鐘，95°C反應15秒，60°C反應1分鐘，共40個循環。基因表現量是由以下公式計算：2 - △ Ct，其中△Ct=Ct 目標基因-Ct β 肌動蛋白。前述黑色素生成相關基因表現之結果如圖4所示。由圖4之結果可知，細胞經龍眼殼萃取液（250 µg/mL）作用後，黑色素生成相關基因MC1R、MITF、TYR、TRP-1和TRP-2的表現量分別為1.4、1.2、4.7、1.6和1.2倍，顯示龍眼殼萃取液藉由調控細胞內黑色素生成相關基因之表現，促進黑色素生成。 It is known that melanin production is regulated by genes such as MC1R, MITF, Tyrosinase, TRP-1 and TRP-2, so the effect of longan shell extract on the expression of melanin production-related genes was further tested. First, melanoma cells (B16 cells) were seeded in 6-well plates at a density of 1×10 5 /mL. After 24 hours of cultivation, the culture solution containing 250 µg/mL longan shell extract was used for 72 hours. After collecting the cells, use RNA extraction reagent (purchased from Protech Company, Taiwan, Cat No. PT-KP200CT) to extract the RNA of the above cells, then use MMLV reverse transcriptase to reverse transcribe the RNA into cDNA, and then use Quantitect SYBR Green RT-PCR The kit (purchased from Qiagen, Germany) was used for quantitative real-time reverse transcription polymerase chain reaction with a fluorescent detection system (prism 7700 sequencing detection system). The conditions were 50°C for 30 minutes and 95°C for 15 minutes , react at 95°C for 15 seconds, and react at 60°C for 1 minute, a total of 40 cycles. Gene expression is calculated by the following formula: 2 - △ Ct , where △Ct=Ct target gene -Ct β- actin . The result of expression of genes related to melanin production is shown in FIG. 4 . It can be seen from the results in Figure 4 that after the cells were treated with longan shell extract (250 µg/mL), the expression levels of melanin production-related genes MC1R, MITF, TYR, TRP-1 and TRP-2 were 1.4, 1.2, 4.7, 1.6 and 1.2 times, showing that the longan shell extract promotes melanin production by regulating the expression of genes related to melanin production in cells.
實施例6 ：龍眼殼頭皮養護產品的製備。Embodiment 6: the preparation of longan shell scalp maintenance product.
已知皮膚具有皮脂膜及角質層，為了使產品中的活性成分順利通過障壁層，達到作用功效，故將其作用濃度提高50-100倍。進行動物模式之塗抹試驗時，文獻上活性成分之建議濃度亦是提高50-100倍。參考實施例2-5之龍眼殼萃取液有效作用濃度為250µg/mL（重量百分濃度0.025%），故於製備龍眼殼頭皮養護產品時，添加重量百分濃度2.5%龍眼殼萃取液及其他成分，包含水、酒精、何首烏萃取液（重量百分濃度0.5%）、褐藻萃取液、土肉桂萃取液、微量元素、PGA、雪松精油、百里香精油、香精。It is known that the skin has a sebum membrane and a stratum corneum. In order to make the active ingredients in the product pass through the barrier layer smoothly and achieve the effect, the concentration of the active ingredient is increased by 50-100 times. When carrying out smear tests on animal models, the recommended concentration of active ingredients in the literature is also increased by 50-100 times. The effective concentration of the longan shell extract in reference examples 2-5 is 250 µg/mL (weight percent concentration 0.025%), so when preparing longan shell scalp care products, add 2.5% weight percent longan shell extract and other Ingredients include water, alcohol, Polygonum multiflorum extract (0.5% by weight), brown algae extract, soil cinnamon extract, trace elements, PGA, cedarwood essential oil, thyme essential oil, and essence.
實施例7 ：龍眼殼頭皮養護產品於頭皮色澤分析。Embodiment 7: Longan shell scalp maintenance products in scalp color analysis.
受試者分別於每日早晨洗臉後及晚上洗澡後，將產品均勻噴抹於頭皮，每次劑量約3-5 mg/cm
2。未使用產品時及使用產品12周後，使用肌膚色差分析儀（Chroma Meter MM500, Minolta, Japan）檢測頭皮色澤，並採用CIE （Commission Internationale deL'Eclariage）system所製訂之標準表色法，以求得頭皮色澤數值。其中，龍眼殼頭皮養護產品於頭皮色澤分析之結果如表1所示，可發現：與未使用本產品之組別相比，於使用該產品十二週後，使用者頭皮皮膚色澤，其明亮度上升至原來的103.6%，表示本產品能夠維護頭皮並保持頭皮健康、有光澤。
表1
實施例8 ：龍眼殼頭皮養護產品於頭皮發紅指數分析。Embodiment 8: Analysis of scalp redness index of longan shell scalp maintenance products.
受試者分別於每日早晨洗臉後及晚上洗澡後，將產品均勻噴抹於頭皮，每次劑量約3-5 mg/cm
2。未使用產品時及使用產品12周後，使用肌膚色差分析儀（Chroma Meter MM500, Minolta, Japan）檢測頭皮色澤，並採用CIE （Commission Internationale deL'Eclariage）system所製訂之標準表色法，以求得頭皮發紅數值。其中，龍眼殼頭皮養護產品於頭皮發紅指數分析之結果如表2所示，可發現：與未使用本產品之組別相比，於使用該產品十二週後，使用者頭皮皮膚發紅指數下降至原來的92.5%，表示本產品可以有效防止頭皮因發炎而產生紅腫之狀況。
表2
實施例9：龍眼殼頭皮養護產品於頭髮粗細分析。Example 9: Analysis of longan shell scalp care products on hair thickness.
受試者分別於每日早晨洗臉後及晚上洗澡後，將產品均勻噴抹於頭皮，每次劑量約3-5 mg/cm
2。未使用產品時及使用產品12周後，取5根頭髮以數位式外徑測微器（Mitutoyo, C/N293-100, Japan）量測頭髮粗細。其中，龍眼殼頭皮養護產品於頭髮粗細分析之結果如表3所示，可發現：與未使用本產品之組別相比，於使用該產品十二週後，使用者之頭髮粗細增加為原本的107.8 %，表示本產品確實有強健髮根之效果。
表3
實施例10：龍眼殼頭皮養護產品於髮根粗細分析。Example 10: Analysis of longan shell scalp care products on hair root thickness.
受試者分別於每日早晨洗臉後及晚上洗澡後，將產品均勻噴抹於頭皮，每次劑量約3-5 mg/cm 2。未使用產品時及使用產品12周後，以顯微鏡放大（100×）觀察髮根粗細狀況，由髮根往上量測頭髮直徑，評估髮根的粗細狀況。其中，龍眼殼頭皮養護產品於髮根顯微觀測分析之結果如圖5所示，可發現：與未使用本產品之組別相比，於使用該產品十二週後，使用者髮根之平均直徑從0.035毫米變為0.042毫米，明顯變粗，表示本產品確實有強健髮根之效果。 After washing the face in the morning and taking a bath in the evening, the subjects sprayed the product evenly on the scalp, with a dose of about 3-5 mg/cm 2 each time. When not using the product and after using the product for 12 weeks, observe the thickness of the hair root with a microscope (100×), measure the diameter of the hair from the root to the top, and evaluate the thickness of the hair root. Among them, the results of microscopic observation and analysis of the hair root of the longan shell scalp care product are shown in Figure 5. It can be found that compared with the group that did not use this product, after using the product for twelve weeks, the hair root of the user The average diameter has changed from 0.035 mm to 0.042 mm, which is obviously thicker, indicating that this product does have the effect of strengthening hair roots.
實施例11 ：龍眼殼頭皮養護產品於頭皮及髮根顯微觀測分析。Embodiment 11: Microscopic observation and analysis of longan shell scalp maintenance products on scalp and hair roots.
受試者分別於每日早晨洗臉後及晚上洗澡後，將產品均勻噴抹於頭皮，每次劑量約3-5 mg/cm2。未使用產品時及使用產品12周後，以多功能舒膚特膚質檢測儀（Soft Plus, Callegari 1930, Italy）放大40倍觀察頭皮及髮根，並拍照記錄頭皮健康狀況及髮根髮色，估使用產品後受試者頭皮健康狀況及髮根髮色狀況。其中，龍眼殼頭皮養護產品於頭皮及髮根顯微觀測分析之結果如圖6所示，可看出於使用該產品之12周後，使用者之頭皮較無紅腫或頭皮屑之情況產生，此外，使用者之髮根顏色變為較深，表示本產品確實有促進黑色素生成之效果。The subjects sprayed the product evenly on the scalp after washing their face in the morning and taking a bath in the evening, with a dose of about 3-5 mg/cm2 each time. When not using the product and after using the product for 12 weeks, observe the scalp and hair root with a multi-functional Soft Plus skin quality detector (Soft Plus, Callegari 1930, Italy) magnified 40 times, and take photos to record the health status of the scalp and hair color of the hair root , Estimate the scalp health status and hair root hair color status of the subjects after using the product. Among them, the results of microscopic observation and analysis of the longan shell scalp care product on the scalp and hair roots are shown in Figure 6. It can be seen that after 12 weeks of using the product, the scalp of the user has no redness or dandruff. In addition, the color of the user's hair roots became darker, indicating that this product really has the effect of promoting melanin production.
惟以上所述者，僅為本發明之較佳實施例，但不能以此限定本發明實施之範圍；故，凡依本發明申請專利範圍及說明書內容所做之簡單的等效改變與修飾，皆仍屬本發明專利涵蓋之範圍內。But what is described above is only a preferred embodiment of the present invention, but it cannot limit the scope of implementation of the present invention; therefore, all simple equivalent changes and modifications made according to the patent scope of the present invention and the contents of the description, All still belong to the scope covered by the patent of the present invention.
無none
圖1為一長條圖，用以表示蕈菇類酪胺酸酶活性抑制率之結果； 圖2為一長條圖，用以表示細胞外黑色素含量影響之結果； 圖3為一長條圖，用以表示細胞內酪胺酸酶活性影響之結果； 圖4為一長條圖，用以表示促進黑色素生成相關基因表現量影響之結果； 圖5為一平面圖，用以表示使用龍眼殼頭皮養護產品後受試者髮根粗細之變化； 圖6為一照片圖，用以表示使用龍眼殼頭皮養護產品後之效果。 Fig. 1 is a long bar graph, in order to represent the result of the activity inhibition rate of mushroom tyrosinase; Fig. 2 is a bar chart, used to represent the result of the influence of extracellular melanin content; Fig. 3 is a bar graph, is used for representing the result of the influence of intracellular tyrosinase activity; Fig. 4 is a bar graph, used for expressing the result that promotes the expression of gene expression related to melanin production; Figure 5 is a plan view showing the changes in the thickness of the hair roots of the subjects after using the longan shell scalp care product; Fig. 6 is a photo graph showing the effect of using the longan shell scalp care product.
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