CN106821948B - 将利用银耳菌丝体的松萝发酵提取物作为有效成分的化妆品组合物 - Google Patents
将利用银耳菌丝体的松萝发酵提取物作为有效成分的化妆品组合物 Download PDFInfo
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- CN106821948B CN106821948B CN201610829133.9A CN201610829133A CN106821948B CN 106821948 B CN106821948 B CN 106821948B CN 201610829133 A CN201610829133 A CN 201610829133A CN 106821948 B CN106821948 B CN 106821948B
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- tremella
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Abstract
本发明涉及一种化妆品组合物,含有松萝银耳菌丝体发酵提取物作为有效成分,该松萝银耳菌丝体发酵提取物是对利用银耳菌丝体使松萝发酵而得的发酵物进行提取而得的,因而可具有改善皱纹、改善皮肤弹性的效果。根据本发明,通过利用银耳菌丝体进行发酵,来明显降低松萝提取物所具有的细胞毒性,因而对皮肤安全,并且,由于具有优异的松萝提取物未曾具有的、骨胶原及弹性蛋白合成功能,因而在用于改善皮肤皱纹及改善皮肤弹性的化妆品组合物方面有利。
Description
技术领域
本发明涉及一种化妆品组合物,该化妆品组合物中,将对利用银耳菌丝体使松萝发酵的发酵物进行提取(extraction)而得的、银耳菌丝体的松萝发酵提取物作为有效成分,因而可具有改善皱纹、改善皮肤弹性的效果。
背景技术
皮肤老化缘于自由基对生物体的氧化性损伤。在生物体内,氧化剂或自由基的攻击和抗氧化剂的防御形成密切的协调。但是,由于抗氧化剂的防御并不是完美无缺,因此使作为生物体组成分子的脂质、蛋白质、核酸、酶等生物体分子持续受到氧化剂或自由基的氧化性损伤,并且其氧化反应产物会随着年龄的增加而蓄积。
尤其,因为皮肤始终与氧相接触,受到暴露于紫外线等的氧化负担,导致基于酶或非酶的抗氧化防御膜破损,引起生物体组成成分的损伤,结果带来皮肤弹性下降、色素沉积、形成皱纹等形态的皮肤老化。所述生物体组成成分的损伤,例如是脂质过氧化、蛋白质氧化、作为弹性纤维的骨胶原和弹性蛋白的链断裂及非正常交叉结合、透明质酸链断裂、黑色素形成反应的促进,脱氧核糖核酸(DNA)氧化等。
尤其,蛋白质的氧化会使骨胶原(collagen)、透明质酸(hyaluronicacid)、弹性蛋白(elastin)、蛋白多糖(proteoglycan)、纤连蛋白(fibronectin)等皮肤结合组织发生断裂并造成严重的多炎症反应以及阻碍皮肤弹性,如果变得更严重,则脱氧核糖核酸发生变异而导致基因突变、诱发癌症、免疫力下降。
为防止这种皮肤老化并提高皮肤弹性,人们使用抗氧化剂、保湿或改善皱纹的化妆品。但是,迄今为止的化妆品仅停留在预防皮肤老化的层面,很难获得实质性的提高弹性的效果。
现有技术文献
专利文献
专利文献1:韩国公开专利第2011-0067994号:含有松萝提取物的皮肤外用剂组合物
专利文献2:韩国公开专利第2007-0055372号:番石榴发酵组合物及其用途
专利文献3:韩国公开专利第2012-0087408号:具有抗氧化活性的银耳发酵提取物及含有其的化妆品组合物
发明内容
对此,在本发明人为开发将天然提取物作为有效成分并具有改善皮肤皱纹及弹性的效果的化妆品组合物而从多方面进行努力的结果,确认到利用银耳菌丝体使松萝发酵,不仅明显降低松萝提取物所具有的细胞毒性,并且还能够发挥松萝提取物未曾具有的、优异的骨胶原及弹性蛋白合成功能,从而显现出对皮肤皱纹以及弹性的改善功效,从而完成了本发明。
因此,本发明的目的在于提供一种化妆品组合物及其制备方法,将利用银耳菌丝体使松萝发酵的发酵物进行提取而得的提取物作为有效成分。
为了实现上述目的,本发明提供一种化妆品组合物,该化妆品组合物中,将对利用银耳菌丝体使松萝发酵获得的发酵物进行提取而得的、银耳菌丝体的松萝发酵提取物作为有效成分。
并且,本发明提供松萝银耳发酵提取物的制备方法,该制备方法包括:对松萝进行灭菌的步骤;使经过灭菌的上述松萝在培养基湿润的步骤;向经过湿润的上述松萝接种银耳菌丝体后进行发酵来获取发酵物的步骤;以及使用提取溶剂对上述发酵物进行提取的步骤。
根据本发明,通过利用银耳菌丝体的发酵,来明显降低松萝提取物所具有的细胞毒性,因而对皮肤安全,并且,由于具有松萝提取物未曾具有的、优异的骨胶原及弹性蛋白合成功能,因而在用于改善皮肤皱纹及皮肤弹性的化妆品组合物方面有利。
附图说明
图1为示出对实施例1至实施例4中的提取物及比较例1中的提取物测定骨胶原合成功能的测定结果的图表。
图2为示出对实施例2中的提取物及比较例1中的提取物测定细胞生存能力的测定结果的图表。
图3为示出对实施例2中的提取物及比较例1中的提取物测定弹性蛋白合成功能的测定结果的图表。
图4为示出在涂抹分别含有实施例2中的提取物及比较例1中的提取物的实验剂型面霜及比较剂型面霜后测定皮肤弹性改善程度的测定结果的图表。
具体实施方式
以下,对本发明进行详细的说明。
本发明公开具有改善皱纹及弹性的效果的、将松萝银耳发酵提取物作为有效成分的化妆品组合物,上述化妆品组合物将松萝作为原料,通过利用银耳菌丝体进行的发酵,明显降低松萝提取物所具有的细胞毒性,使改善皮肤皱纹及弹性的效果最大化。
松萝(Usnea longissima)是呈黄色或绿色的灌木状地衣类植物,具有长茎和缠绕在一起的线团般的圆盘形状附着器。中医中用于利尿剂、解热剂、祛痰剂。生长区域分布在温带地区的高山至寒带,分布在韩国、日本等地。这种松萝提取物以抑制黑色素的生成、增加透明质酸来具有美白皮肤、防止皮肤老化的效果而闻名。
本发明利用银耳菌丝体使松萝发酵。
银耳是担子菌类非褶菌目银耳科的蘑菇,生长在韩国、日本、欧洲、北美洲、澳大利亚等地,主要生长在针叶树的桩子或枯木周边。其直径为10~30cm,子实体高为10~25cm,肉质,底部粗壮的柄上发出许多分枝,枝端形成无数曲折的瓣片,形似巨大的绣球。因100g的银耳中含有43.6g的β-葡聚糖而具有出色的抗癌效果,此外还以降血压或将血糖恢复正常并具有造血功能而闻名。
具体地,本发明的松萝银耳发酵提取物的制备方法包括:对松萝进行灭菌的步骤;使经过灭菌的上述松萝在培养基湿润的步骤;向经过湿润的上述松萝接种银耳菌丝体后进行发酵来取得发酵物的步骤;以及使用提取溶剂对上述发酵物进行提取的步骤。
以下,对各步骤进行详细的说明。
首先,对松萝进行灭菌。
灭菌是利用高压蒸气灭菌器(autoclave)对松萝中存在的所有菌株进行灭菌,在业界常规条件下进行。作为一例,通过在100℃至125℃的温度下进行5分钟至30分钟的热处理来实施对松萝进行灭菌的步骤。
接着,使经过灭菌的上述松萝在培养基湿润。
通过使松萝在培养基中湿润,在之后的利用银耳菌丝体进行发酵时,供给菌丝体的碳源、能源,从而更加有效地进行发酵。
上述培养基可选自由马铃薯葡萄糖琼脂(potato dextrose agar)、马铃薯葡萄糖肉汤(potato dextrose broth)、沙氏葡萄糖肉汤(sabouraud dextrose broth)及它们的混合物组成的组中的一种。
相对于培养基重量,向上述培养基添加的松萝的含量为1.0重量%至30重量%。
接着,向经过湿润的上述松萝接种银耳菌丝体后进行发酵来获得发酵物。
此时,发酵可在适宜的菌丝体生长环境下实施。优选地,在20~30℃的温度下实施发酵,更为优选地,在25~30℃的温度下通过10天至30天的静置培养实施发酵。若发酵温度小于20℃,则菌丝体不容易生长,若发酵温度大于30℃,则菌丝体的生长过快,有可能对提取物的性能有所影响。
最后,使用提取溶剂对上述发酵物进行提取。
可使用的提取溶剂为选自由水、碳数1至6的多元醇、碳数1至4的低级醇、乙酸乙酯、甘油及它们的混合溶剂组成的组中的1种,优选地,使用70%乙醇,添加达到原料重量的2~50倍(体积)的提取溶剂。
上述提取可使用热水提取、常温提取、高压提取、加温提取、超声波提取或超临界提取等常规方法。
提取温度可以由技术人员根据提取方法采用适当、多样的温度范围,例如,可在20℃至100℃的温度下进行提取,但并不局限于此。并且,提取时间根据提取方法而不同,技术人员可采用适当的提取时间,但并不局限于此,可在约1小时至数日的范围内实施单次或数次提取。对于上述利用第1次提取溶剂进行提取并获得的提取物,可以使用常规方法进行过滤而获得去掉杂质的液状提取物,或可使用常规方法对所得的液状提取物进行减压浓缩和/或干燥而获得粉末形态的提取物。
相对于组合物总重量,含有0.0001重量%至90重量%的本发明的上述提取物,优选含有0.001重量%至30.0重量%的本发明的上述提取物。为实现最低程度的改善皮肤弹性及皱纹的效果,优选提取物的含量为上述最小值以上,考虑到因大量添加而导致的使用感下降及对各种剂型的适用可能性,优选提取物的含量为上述最大值以下。此时,优选根据剂型中含有的成分含量来在上述范围内适当调节提取物的含量。
并且,在将本发明的提取物用作组成成分时,能够以冷冻干燥的粉末状态进行使用,不仅如此,还能够以分散或溶解在水或常规有机溶剂中的液体状态进行使用。
并且,优选在本发明化妆品组合物中,在不损伤本发明主要效果的前提下,除上述提取物成分以外,可添加对主要效果起提升作用的其他成分。
本发明的化妆品组合物所包含的成分除上述有效成分以外,还可包含通常用于化妆品组合物的成分,例如,可包含稳定剂、增溶剂、维生素、颜料及香料等常规助剂和载体。
本发明的化妆品组合物可制备成本领域中通常使用的任何剂型,具体可制备成溶液、悬浮液、乳浊液、胶、乳液、精华素、面霜、粉饼、香皂、面膜、洗面奶、油、粉底、喷射剂等化妆品剂型。
在本发明的剂型为溶液或乳浊液的情况下,作为载体成分可使用溶剂、增溶剂或乳浊化剂,例如有水、乙醇、异丙醇、碳酸乙酯、乙酸乙酯、苯甲醇、苯甲酸苄酯、丙二醇、1,3-丁基乙二醇油、甘油脂肪酯、聚乙二醇或脱水山梨糖醇的脂肪酸酯。
在本发明的剂型为悬浮液的情况下,作为载体成分可使用如水、乙醇或丙二醇等液状稀释剂、乙氧基化异硬脂醇、聚氧乙烯山梨糖醇酯及聚氧乙烯山梨糖醇酐酯等的悬浮剂、羟基氧化铝、微晶纤维素、膨润土、琼脂或黄蓍树胶等。
在本发明的剂型为面霜或胶的情况下,作为载体成分可使用蜡、石蜡、黄蓍树胶、动物性油、淀粉、纤维素衍生物、硅酮、膨润土、聚乙二醇、二氧化硅、氧化锌或滑石粉等。
在本发明的剂型为粉饼或喷射剂的情况下,作为载体成分可包含如二氧化硅、滑石粉、氢氧化铝、乳糖、硅酸钙、氯氟烃、丙烷/丁烷或二甲醚等促进剂。
在本发明的剂型为含表面活性剂的洗面奶的情况下,作为载体成分可使用脂肪醇硫酸盐、脂肪醇醚硫酸盐、磺基琥珀酸单酯、咪唑啉鎓衍生物、羟乙基磺酸盐、甲基牛磺酸盐、肌氨酸盐、脂肪酸酰胺醚硫酸盐、脂肪族醇、烷基酰胺甜菜碱、脂肪酸甘油酯、脂肪酸二乙醇酰胺、植物性油、羊毛脂衍生物或乙氧基化甘油脂肪酸酯等。
以下,为了有助于理解本发明而公开优选实施例。但是,以下实施例仅用于更加容易地理解本发明,本发明的内容并不限制于实施例。
实施例1至实施例4:制备利用银耳菌丝的松萝发酵提取物
在向沙氏葡萄糖肉汤培养基(sabouraud dextrose broth)中添加经灭菌的松萝8wt%后湿润。之后,相对于湿润的松萝,接种30wt%的经培养的银耳菌丝,在30℃的温度下分别静置培养5天、10天、20天、30天。将100g的所获得的发酵物放入重量比达20倍的70%乙醇并在25℃的温度下提取4个小时后过滤,除去菌体等不溶性物质,利用旋转减压浓缩机除去了溶剂。所获得的松萝发酵提取物的收率如下表1。
表1
区分 | 实施例1 | 实施例2 | 实施例3 | 实施例4 |
发酵时间(day) | 5 | 10 | 20 | 30 |
收率(%) | 3.22 | 3.14 | 1.11 | 3.42 |
比较例1:松萝提取物
粉碎200g的松萝并放入重量比达20倍的70%乙醇中,在25℃的温度下提取4个小时后过滤,除去菌体等不溶性物质,利用旋转减压浓缩机除去了溶剂,获得了松萝提取物。
实验例1:按发酵时间比较松萝发酵提取物的骨胶原合成功能
按每孔形成1×106细胞,在48孔微型板接种作为人体正常上皮细胞的纤维芽细胞,并在DMEM培养基中培养24小时。以100μg/ml添加阳性对照组及实施例1至实施例4、比较例的提取物,补充培养48小时。培养后,利用骨胶原试剂盒(宝生物工程,Takara,日本)对各孔的上层液测定IC型前胶原(procollagen)钛(PIP)的量,测定了由此合成的骨胶原的量。图1及表2示出其结果。
表2
区分 | PIP-1生成量(%) |
对照组 | 100 |
阳性对照组(EGF) | 116.3 |
实施例1 | 81.7 |
实施例2 | 146.6 |
实施例3 | 125.7 |
实施例4 | 138.7 |
比较例1 | 88.1 |
参照图1及表2的结果,相对于阳性对照组,本发明的松萝发酵提取物呈现出优异的骨胶原合成功能。尤其,发酵10天的实施例2的松萝发酵提取物呈现出最优异的效果。这可以认为通过利用银耳菌丝的发酵过程,转换为可呈现出未曾在松萝提取物中确认到的骨胶原合成功能的有效成分。
实验例2:确认通过发酵减少毒性
为了比较在上述实施例2及比较例1中获得的样本的细胞生存力,应用以下文献中的MTT assay方法进行了实验(Han HK、Choi SS、Shin JC<Chung HS(2007)A study onsingle oral dose toxicity of highlydeveloped anthocyanin-pigmented ricevarieties.J.Kor.Soc.Food Sci.Nutr.36:527-533)。对照组(control)是未处理组,以对照组的细胞生存率作为100%,来示出了结果。图2表示在多种浓度(10μg/ml、25μg/ml、50μg/ml、100μg/ml、200μg/ml)条件下确认到的比较例1及实施例2的细胞生存率结果。
如图2表示,以200μg/ml浓度处理时,实施例2的松萝发酵提取物呈现出了约80%的细胞生存率,相反,在比较例1的松萝提取物中确认到约10%左右的细胞生存率。由此可以确认,通过发酵,松萝提取物的细胞毒性减少约2倍,由此,上述松萝发酵提取物可以作为对皮肤安全的有效成分来用于化妆品。
实验例3:确认松萝提取物和松萝发酵提取物的弹性蛋白合成功能
将纤维芽细胞(CCD-986Sk)利用添加有10%FBS的IMDM培养基调节为5×105cell/well后,接种到12孔板培养了18个小时。培养后,以100μg/ml浓度对上述实施例2及比较例1的样本进行处理并培养了24个小时。从所培养的细胞中提取核糖核酸(RNA)并通过反转录-聚合酶链式反应(RT-PCR,Reverse transcription polymerase chain reaction)确认了弹性蛋白原(tropoelastin)的表达增量。图3表示了其结果。
参照图3,可知通过发酵可增强弹性蛋白合成功能。
实验例4:确认皮肤弹性度的改善
根据表3,通过常规方法制备含有实施例2的松萝发酵提取物的面霜,并确认了弹性改善度。
表3
利用上述比较剂型及实验剂型,对年龄在30岁至40岁之间的10名女性作为实验对象,按一天两次在皮肤上持续涂抹,两周后,利用cutometer MPA 280设备确认了皮肤弹性。根据数学式1计算其改善率并在图4中进行示出。如图4表示,确认到通过发酵可以增强皮肤弹性改善率。
数学式1
弹性改善率(%)=(剂型使用组弹性一对照组弹性)×100/对照组弹性
以下,以本发明的剂型例说明柔肤化妆水、收敛化妆水、营养化妆水、营养面霜、按摩面霜、精华素及面膜,但本发明的组合物剂型并不局限于此。
剂型例1:柔肤化妆水(化妆水)
根据表4,以常规方法制备了含有实施例2的松萝发酵提取物的柔肤化妆水。
表4
成分 | 含量(%) |
实施例2的松萝发酵提取物 | 1 |
甘油 | 5 |
1,3-丁二醇 | 3 |
甜菜碱 | 1 |
尿囊素 | 0.1 |
DL-泛醇 | 0.3 |
EDTA-2Na | 0.02 |
透明质酸钠粉 | 0.05 |
乙醇 | 5 |
辛基十二醇聚醚-16 | 0.2 |
聚氧乙烯硬化蓖麻油 | 0.2 |
香料 | 适量 |
防腐剂 | 适量 |
色素 | 适量 |
纯净水 | 余量 |
合计 | 100 |
剂型例2:营养化妆水(乳液(lotion))
根据表5,通过常规方法制备了含有实施例2的松萝发酵提取物的营养化妆水。
表5
成分 | 含量(%) |
实施例2的松萝发酵提取物 | 3 |
甘油硬脂酸酯SE | 1.5 |
鲸蜡硬脂醇 | 1.0 |
牛油树脂 | 1.5 |
聚山梨醇酯60 | 1.3 |
去水山梨糖醇单硬脂酸酯 | 0.5 |
硬化食物油 | 1 |
矿物油 | 5 |
角鲨烷 | 3 |
环甲硅酮 | 2 |
二甲基硅酮 | 0.8 |
生育酚醋酸酯 | 0.5 |
卡波姆 | 0.12 |
甘油 | 5 |
1,3-丁二醇 | 3 |
透明质酸钠粉 | 0.05 |
三乙醇胺 | 0.12 |
香料 | 适量 |
防腐剂 | 适量 |
色素 | 适量 |
蒸馏水 | 余量 |
合计 | 100 |
剂型例3:按摩霜
根据表6,通过常规方法制备了含有实施例2的松萝发酵提取物的按摩霜。
表6
成分 | 含量(%) |
实施例2的松萝发酵提取物 | 3 |
亲油性一硬脂酸甘油酯 | 1.5 |
鲸蜡硬脂醇 | 1.5 |
硬脂酸 | 1 |
聚山梨醇酯60 | 1.5 |
去水山梨糖醇硬脂酸酯 | 0.6 |
异硬脂酸异硬脂醇酯 | 5 |
角鲨烷 | 5 |
矿物油 | 35 |
二甲基硅酮 | 0.5 |
羟乙基纤维素 | 0.12 |
甘油 | 6 |
1,3-丁二醇 | 3 |
三乙醇胺 | 0.3 |
香料 | 适量 |
防腐剂 | 适量 |
色素 | 适量 |
蒸馏水 | 余量 |
合计 | 100 |
剂型例4:精华素
根据表7,通过常规方法制备了含有实施例2的松萝发酵提取物的精华素。
表7
成分 | 含量(重量%) |
实施例2的松萝发酵提取物 | 3 |
甘油 | 6 |
甜菜碱 | 5 |
PEG 1500 | 2 |
尿囊素 | 0.1 |
DL-泛醇 | 0.3 |
EDTA-2Na | 0.02 |
氢化卵磷脂 | 0.6 |
羟乙基纤维素 | 0.1 |
透明质酸钠粉 | 0.08 |
羧基乙烯基聚合物 | 0.2 |
三乙醇胺 | 0.2 |
神经酰胺 | 0.2 |
辛基十二醇 | 3 |
角鲨烷 | 3 |
聚山梨醇酯60 | 0.4 |
甘油硬脂酸酯SE | 1.5 |
香料 | 适量 |
防腐剂 | 适量 |
色素 | 适量 |
蒸馏水 | 余量 |
合计 | 100 |
剂型例5:面膜
根据下表8,通过常规方法制备了含有实施例2的松萝发酵提取物的面膜。
表8
成分 | 含量(重量%) |
实施例2的松萝发酵提取物 | 1 |
聚乙烯醇 | 15 |
羧甲基纤维素 | 0.15 |
甘油 | 3 |
PEG 1500 | 2 |
甜菜碱 | 2 |
DL-泛醇 | 0.4 |
尿囊素 | 0.1 |
三乙醇胺 | 0.2 |
烟酰胺 | 0.5 |
乙醇 | 6 |
PEG 40硬化蓖麻油 | 0.3 |
香料 | 适量 |
防腐剂 | 适量 |
色素 | 适量 |
蒸馏水 | 余量 |
合计 | 100 |
Claims (6)
1.一种化妆品组合物,含有对发酵物使用乙醇进行提取而得的提取物作为有效成分,所述发酵物为利用银耳菌丝体,使松萝在沙氏葡萄糖肉汤培养基,在20℃~30℃的温度下进行10天~30天的发酵而得的发酵物,其中相对于化妆品组合物的总重量,上述提取物的含量为1重量%~3重量%。
2.根据权利要求1所述的化妆品组合物,其中上述提取物具有液状、浓缩的固状或冷冻干燥的粉末形态。
3.根据权利要求1所述的化妆品组合物,其中上述提取物具有改善皮肤皱纹的效果。
4.根据权利要求1所述的化妆品组合物,其中上述提取物具有改善皮肤弹性的效果。
5.根据权利要求1~4中任一项所述的化妆品组合物,其中上述化妆品组合物具有选自由溶液、悬浮液、乳浊液、胶、乳液、精华素、面霜、粉、香皂、面膜、洗面奶、油、粉底、喷雾组成的组中的1种剂型。
6.一种松萝银耳发酵提取物的制备方法,其中包括:
对松萝进行灭菌的步骤;
使经过灭菌的上述松萝在沙氏葡萄糖肉汤培养基湿润的步骤;
向经过湿润的上述松萝接种银耳菌丝体后,在20℃~30℃的温度下进行10天~30天的发酵,来取得发酵物的步骤;以及
使用乙醇作为提取溶剂对上述发酵物进行提取的步骤。
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