CN105434201A - 含石榴提取物的组合物及其用途和皮肤外用剂 - Google Patents
含石榴提取物的组合物及其用途和皮肤外用剂 Download PDFInfo
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- CN105434201A CN105434201A CN201410499969.8A CN201410499969A CN105434201A CN 105434201 A CN105434201 A CN 105434201A CN 201410499969 A CN201410499969 A CN 201410499969A CN 105434201 A CN105434201 A CN 105434201A
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- Prior art keywords
- extract
- punica granatum
- skin preparations
- extenal use
- compositions
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Abstract
本发明公开了一种含石榴提取物的组合物及其用途和一种皮肤外用剂。该含石榴提取物的组合物包括下述质量百分比的组分:石榴提取物0.001-4.7%和冰川水95.3-99.999%。本发明的含石榴提取物的组合物能够显著地抑制HEM-m细胞黑色素的生成,并且天然、安全,可用作皮肤外用剂的美白活性成分。本发明的皮肤外用剂可以有化妆水、精华液、乳液、乳霜等多种形式,其成分天然、安全,并具有美白的功效。
Description
技术领域
本发明涉及一种含石榴提取物的组合物及其用途和一种皮肤外用剂。
背景技术
石榴树(PunicagranatumL.)为落叶乔木或灌木,浆果球形,顶端有宿存花萼裂片,果皮厚;种子多数,浆果近球形,果熟期9-10月。外种皮肉质半透明,多汁;内种皮革质。石榴性味甘、酸涩、温,具有杀虫、收敛、涩肠、止痢等功效。
冰川水是地表上长期存在并能自行运动的天然冰体。由大气固体降水经多年积累而成,是地表重要的淡水资源,不同于冬季河湖冻结的水冻冰。新雪降落到地面后,经过一个消融季节未融化的雪叫粒雪。雪逐步密实、经融化再冻结,使晶体合并,晶粒改变其大小和形态,出现定向增长。当其密度达到临界值,晶粒间推动透气性和透水性,便成为冰川冰。许多国家的科学家已经对常喝冰川水有利于提高人类体质、减少疾病、延长寿命等现象进行了系列研究。
冰川水的珍贵,被中外科学界称为“生命之水”。而取水点的不同,深刻影响着冰川水对生命的神奇作用。纵向分析,位于雪线以下地质深层处万年老冰融水处的冰川融水纯度最高,而越往下的冰川融水,一定程度上汇聚新年份冰雪融水甚至生物影响,纯净程度不断减弱;横向分析,山之阴的冰川优于山之阳。水蒸气中较重分子在顺山体上升过程中受重力作用,不断形成降雨降雪落于山间,爬过山顶落至山阴的将是经自然过滤后最轻、最纯净的水体。
发明内容
本发明所解决的技术问题在于提供了一种含石榴提取物的组合物及其用途和一种皮肤外用剂。本发明的含石榴提取物的组合物能够显著地抑制HEM-m细胞黑色素的生成,同时表现出优良的保湿性,并且天然、安全,可用作皮肤外用剂的美白活性成分、保湿活性成分,本发明的皮肤外用剂成分天然、安全,并具有美白、保湿的功效。
本发明通过以下技术方案解决上述技术问题。
本发明提供了一种含石榴提取物的组合物,其包括下述质量百分比的组分:石榴提取物0.001-4.7%和冰川水95.3-99.999%。
其中,所述的含石榴提取物的组合物较佳地包括下述质量百分比的组分:石榴提取物0.001-4.5%和冰川水95.5-99.999%。
其中,所述的石榴提取物可为本领域常规使用的石榴提取物。本发明中,所述的石榴提取物较佳地由下述制备方法制得:取酸石榴,分离其皮、籽部位,粉碎至粗粉,所述粗粉的粒径为40-100目,加8-12倍体积50-90%乙醇超声提取2-3次,每次1-3小时,合并滤液,浓缩,经AB-8大孔吸附树脂富集,依次以水、30%乙醇和70%乙醇洗脱,收集70%乙醇洗脱部位,浓缩至干,得石榴提取物。
更佳地,所述的石榴提取物由下述制备方法制得:取云南玉溪产酸石榴,分离其皮、籽部位,石榴皮粉碎至粗粉,所述粗粉的粒径为40-100目,加8-12倍体积50-90%乙醇超声提取2次,每次2小时,合并滤液,浓缩,经AB-8大孔吸附树脂富集,依次以水、30%乙醇和70%乙醇洗脱,收集70%乙醇洗脱部位,浓缩至干,得石榴提取物。
其中,所述冰川水的水源地可为任意地区,较佳地为喜马拉雅山。所述冰川水的取水口较佳地为多吉曲登尼玛泉。多吉曲登尼玛泉位于喜马拉雅山北坡,海拔5128米,其泉口温度常年保持在1.5-2℃。
本发明中,所述的冰川水较佳地满足下述条件:所述冰川水的pH值为7.1-7.9,所述冰川水中的氘含量为133-137ppm,且所述冰川水中不含溴酸盐;更佳地,每升所述冰川水含有下述组分:Sr+0.2-0.5mg、K+0.5-10.0mg、Na+1.0-5.0mg、Ca2+14.0-30.0mg以及Mg2+2.0-10.0mg;所述冰川水中的溶解性总固体含量为60-150mg/L;最佳地,所述冰川水的pH值为7.4,所述冰川水中的氘含量为133ppm。
在本发明中,所述含石榴提取物的组合物的制备方法是将所述的石榴提取物和所述的冰川水混合即可。
本发明还提供了所述含石榴提取物的组合物的用途,所述含石榴提取物的组合物作为皮肤外用剂中的美白活性成分和/或保湿活性成分。
具体地,所述的美白活性成分为抑制人源表皮黑色素细胞HEM-m的黑色素生成的美白活性成分。
本发明还提供了一种皮肤外用剂,其包括所述的含石榴提取物的组合物。
其中,所述含石榴提取物的组合物在所述皮肤外用剂中的含量较佳地为65.1-89.2wt%。
其中,所述含石榴提取物的组合物较佳地由下述质量百分比的组分组成:所述石榴提取物0.001-4.5%和所述冰川水95.5-99.999%。
本发明中,所述皮肤外用剂是通常用于皮肤外部的所有成分的统称概念,例如可以是化妆品或药品。所述化妆品中可以是基础化妆品、面部妆容化妆品、身体用化妆品、头部用护理品等,对其剂型无特殊限制,根据不同目的可合理选择。
本发明中,所述的皮肤外用剂还可以包含其他活性成分,所述其他活性成分较佳地为保湿活性成分、抗氧化活性成分、控油活性成分、美白活性成分和抗衰老活性成分中的一种或多种。所述其他活性成分的含量为本领域常规的含量。
其中,所述的保湿活性成分包括天然保湿活性成分和/或合成保湿活性成分,所述天然保湿活性成分可以选自:角鲨烷、霍霍巴油、蜂蜜、灵芝提取液、芦荟提取物、透明质酸、神经酰胺、丝蛋白类保湿活性成分、胶原蛋白等天然物质;所述合成保湿活性成分可以选自:多元醇类保湿成分(甘油、丙二醇、丁二醇、戊二醇、聚乙二醇、聚甘油等)、乳酸钠、葡萄糖衍生物、聚丙烯酸树脂、蓖麻油及其衍生物、甲壳素壳聚糖及其衍生物、吡咯烷酮羧酸钠等化学合成物。
其中,所述的抗氧化活性成分可以选自:黄酮类抗氧化活性成分(水飞蓟素、黄芪苷、芦丁、槲皮素、青兰苷、双氢青兰苷、芒果苷等),鞣质类抗氧化活性成分(表没有食子儿茶素、表没有食子儿茶素没食子酸酯、表儿茶素没食子酸酯、儿茶素等),醌类抗氧化活性成分(紫草素等),二肽类氨基酸(蛋氨酸、色氨酸、苯丙氨酸、脯氨酸等),糖醇类抗氧化活性成分(五碳糖、六碳糖单糖、百合多糖、果糖、糖醇等),多酚类抗氧化活性成分(谷物的胚芽、葡萄酚、迷迭香酚、表迷迭香酚、异迷迭香酚等),生物碱类抗氧化活性成分(川穹嗪、药根碱、木兰碱、海罂粟碱、潘荔枝碱等),维生素类抗氧化活性成分(维生素E、维生素C、类胡萝卜素)和植物提取物类的抗氧化活性成分,如凹顶藻提取物等。
其中,所述的控油活性成分可以选自:收敛剂(氧化锌、硫酸锌、氯化铝、硫酸铝、单宁酸、柠檬酸、乳酸等),清凉剂(酒精和薄荷),维生素类控油活性成分(维生素B6、维生素B3和维生素H)和角质剥离溶解剂(水杨酸、果酸等)。
其中,所述的美白活性成分可以选自:熊果苷及其衍生物、曲酸及其衍生物、壬二酸及其衍生物、亚油酸及其衍生物、维生素C及其衍生物,以及果酸。
其中,所述的抗衰老活性成分可以选自:维生素类抗衰老活性成分(维生素A、维生素B),酶(过氧化歧化酶SOD、谷胱甘肽过氧化酶GPO、过氧化氢酶等)和植物提取物(刺山柑芽提取物、飞机草提取物等)。
按照本领域常识,根据不同的剂型和目的,所述的皮肤外用剂中还可以含有本领域中常规使用的介质或基质赋形剂,只要所述赋形剂为化妆品或药学领域中可接受的赋形剂。所述赋形剂的含量为本领域常规的含量。
本发明中,所述赋形剂的形式可以为水相、油相、凝胶、水包蜡型乳液、水包油型乳液或油包水型乳液的形式。所述水相可以为一种或多种水溶性或分散性组分的混合物,其在室温下可以为液体、半固体或固体。本领域技术人员可以基于本领域技术人员掌握的知识选择合适的赋形剂的形式,以及其中包含的组分。
当所述赋形剂的形式为水相时,所述赋形剂较佳地包括水,或水与至少一种亲水性有机溶剂的混合物。所述的亲水性有机溶剂诸如醇,尤其是含有2-5个碳原子的直链或支链低级一元醇,如乙醇或丙醇;多元醇,如丙二醇、山梨醇、甘油、泛醇或聚乙二醇及其混合物。
当所述赋形剂的形式为乳液时,所述的皮肤外用剂较佳地还可以包括流变改进剂和/或成膜剂。所述流变改进剂可以选自:高分子增稠剂;疏水改性的丙烯酸酯共聚物,例如丙烯酸酯/Steareth-20丙烯酸甲酯共聚物、丙烯酸酯/Beheneth-25丙烯酸酯共聚物;疏水改性的乙氧基化聚胺酯;例如PEG-150/癸醇/SMDI共聚物、PEG-150/硬脂醇/SMDI;疏水改性非离子多羟基化合物,例如PEG-150二硬脂酸盐。
所述高分子增稠剂可以选自:纤维素类增稠剂、聚丙烯酸类增稠剂、天然胶及其改性物、无机高分子及其改性物、聚氧乙烯类增稠剂和其它类增稠剂;其中,纤维素类增稠剂可以选自:纤维素、纤维素胶、羧甲基羟乙基纤维素、鲸蜡羟乙基纤维素、乙基纤维素、羟乙基纤维素、羟丙基纤维素、羟丙基甲基纤维素、甲基纤维素、羧甲基纤维素等;聚丙烯酸类增稠剂可以选自:丙烯酸酯/C10-30烷基丙烯酸酯交联聚合物、丙烯酸酯/十六烷基乙氧基(20)衣康酸酯共聚物、丙烯酸酯/十六烷基乙氧基(20)甲基丙烯酸酯共聚物、丙烯酸酯/十四烷基乙氧基(25)丙烯酸酯共聚物、丙烯酸酯/十八烷基乙氧基(20)衣康酸酯共聚物、丙烯酸酯/十八烷基乙氧基(20)甲基丙烯酸酯共聚物、丙烯酸酯/十八烷基乙氧基(50)丙烯酸酯共聚物、丙烯酸酯/VA交联聚合物、丙烯酸钠/乙烯异癸酸酯交联聚合物、聚丙烯酸及其钠盐等;天然胶及其改性物可以选自:海藻酸及其(铵、钙、钾)盐、果胶、透明质酸钠、瓜尔胶、黄原胶、阳离子瓜尔胶、羟丙基瓜尔胶、黄蓍胶、鹿角菜胶及其(钙、钠)盐、汉生胶、菌核胶等;无机高分子及其改性物可以选自:硅酸铝镁、二氧化硅、硅酸镁钠、水合二氧化硅、蒙脱土、硅酸锂镁钠、水辉石、硬脂铵蒙脱土、硬脂铵水辉石、季铵盐-90蒙脱土、季铵盐-18蒙脱土、季铵盐-18水辉石等;聚氧乙烯类增稠剂可以选自:PEG-n(n=5M、9M、23M、45M、90M、160M)等;其它类增稠剂可以选自:聚乙烯甲基醚/丙烯酸甲酯与癸二烯的交联聚合物、聚乙烯吡咯烷酮等。
所述成膜剂可为本领域常规使用的成膜聚合物,只要其可溶于或分散于所述赋形剂中。所述成膜剂较佳地为下述中的一种或多种:聚氨基甲酸酯、聚丙烯酸均聚物或共聚物、聚酯、基于烃的树脂和/或硅氧烷树脂。
当所述赋形剂的形式为油相时,所述赋形剂较佳地包括在室温下为液体的油溶性或油分散性组分和/或在室温下为油状或蜡状的物质。
所述在室温下为液体的油溶性或油分散性组分较佳地包括下述物质中的一种或多种:来源于动物的基于烃的油,如全氢化角鲨烯;基于烃的植物油,如液体的C4-10脂肪酸的甘油三酯类,例如庚酸甘油三酯类、辛酸甘油三酯类、癸酸甘油三酯类,或植物油,例如向日葵油、玉米油、大豆油、葡萄籽油、蓖麻油、鳄梨油、霍霍巴油;矿物或合成来源的直链或支链烃类,例如液体石蜡及其衍生物、凡士林;合成酯类和醚类,特别是脂肪醇的酯类,例如肉豆蔻酸异丙酯、棕榈酸2-乙基己酯、硬脂酸2-辛基十二烷基酯、异硬脂酸异硬脂醇酯;羟基化酯类,例如乳酸异硬脂醇酯、羟基硬脂酸辛酯、羟基硬脂酸辛酯、羟基硬脂酸辛基十二烷基酯、脂肪醇的庚酸酯类、辛酸酯类和癸酸脂类;多元醇酯类,例如丙二醇二辛酸酯、新戊二醇二庚酸酯、二甘醇二异壬酸酯和季戊四醇酯类;含有C12-26的脂肪醇类,例如辛基十二烷醇、2-丁基辛醇、2-己基癸醇、2-十一烷基十五烷醇、油醇;基于部分烃的氟油和/或氟硅油,硅油,在室温下为液体或半固体的挥发性或非挥发性的直链或环状聚甲基硅氧烷,例如环状聚二甲基硅氧烷和聚二甲基硅氧烷,其任选包含苯基,例如苯基三甲基硅氧烷、硅氧烷及其混合物。
所述在室温下为油状或蜡状的物质较佳地为蜡和/或树胶。所述的蜡较佳地包括下述物质中的一种或多种:基于烃的蜡、氟蜡和/或硅氧烷蜡,并且可以来源于植物、矿物、动物和/或合成来源;所述的蜡更佳地包括下述物质中的一种或多种:蜂蜡、巴西棕榈蜡、小烛树蜡、石蜡、微晶蜡、地蜡、聚乙烯蜡、含有C16-45的硅氧烷蜡。所述的树胶一般为聚二甲基硅氧烷或羧甲基纤维素钠或多糖类,并且半固体物质一般为基于烃的化合物,如羊毛脂及其衍生物。
本发明的皮肤外用剂还可以进一步包含常规的用于化妆品领域中的任何其他组分,例如防腐剂、芳香剂以及亲水性和亲脂性活性剂中的一种或多种。该些其他组分的含量可为本领域常规的含量。
本发明中,所述皮肤外用剂还可以包含颗粒相,所述的颗粒相可以为化妆品组合物中使用的颜料、珠光剂和填充剂中的一种或多种。所述颗粒相的含量可为本领域常规的含量。所述颜料较佳地为无机颜料和/或有机颜料。所述无机颜料较佳地包括氧化钛、氧化锆和氧化铈、氧化锌、氧化铁和铁蓝中的一种或多种。所述有机颜料较佳地包括钡色淀、锶色淀、钙色淀和铝色淀和碳黑。所述珠光剂较佳地为涂覆了氧化钛、氧化铁或天然颜料的云母。所述填充剂较佳地包括滑石粉、二氧化硅、硬脂酸锌、云母、高岭土、尼龙粉末、聚乙烯粉末、特氟龙、淀粉、一氮化硼和共聚物微球中的一种或多种。所述的共聚物微球较佳地为硅氧烷树脂微珠。
本发明的皮肤外用剂还可以进一步包括一种或多种下述成分:抗过敏剂、抗微生物剂、抗氧化剂、螯合剂、着色剂去色素剂、润肤剂、乳化剂、表皮脱落剂、香料、保湿剂、昆虫驱避剂、润滑剂、药物活性剂、增湿剂、耐光剂、防腐剂、护肤剂、皮肤渗透增强剂、防晒剂、稳定剂、表面活性剂、增稠剂、粘度调节剂、维生素或其任意组合。该些成分的含量可为本领域常规的含量。
本发明中,所述的皮肤外用剂的形式可以为任何合适的产品形式,所述的产品形式包括,但不限于气溶胶型喷雾剂、霜剂、乳液、固体、液体、分散体、泡沫、凝胶、化妆水、摩丝、软膏、粉剂、贴剂、润发油、手按泵型喷雾剂、棒状物、面膜和湿纸巾。众所周知地,本发明的皮肤外用剂可为化妆品、皮肤病学或药物局部施用产品,其制备方法可为本领域常规的制备方法。
在本发明的较佳实施方式一中,所述的皮肤外用剂为化妆水,所述的皮肤外用剂包含下述重量百分比的组分:所述含石榴提取物的组合物84.2-89.2%,甘油5%,多元醇类A3-6%,甜菜碱1%,泛醇0.5%,甘草酸二钾0.05-0.1%,其他活性成分A0.11-2.0%,聚乙二醇-320.5%,表面活性剂A0.15-0.2%,防腐剂A0.4-0.45%,黄原胶0-0.15%和香料0.03%;其中,所述含石榴提取物的组合物中,所述石榴提取物的含量为0.1-3.6wt%,所述冰川水的含量为96.4-99.9wt%;所述的多元醇类A为丁二醇、1,2-戊二醇和双丙甘醇中的一种或多种;所述其他活性成分A为尿囊素、烟酰胺和透明质酸钠中的一种或多种;所述表面活性剂A为羟乙基纤维素、聚山梨醇酯-20和PEG-40氢化蓖麻油中的一种或多种;所述防腐剂A为防腐剂IS-45、羟苯甲酯和苯氧乙醇中的一种或多种。
按照本领域常识,防腐剂IS-45为美国Germall公司生产的防腐剂,其中含有丙二醇、双咪唑烷基脲、羟苯甲酯和碘代丙炔基丁基甲胺酸。
在本发明的较佳实施方式二中,所述的皮肤外用剂为精华液,所述皮肤外用剂包含氨甲基丙醇,以及下述重量百分比的组分:所述含石榴提取物的组合物82.4-85.6%,甘油5%,多元醇类B3-6%,泛醇1%,甘草酸二钾0.1%,尿囊素0.1%,其他活性成分B2.1-4.3%,表面活性剂B0.05-3.6%,防腐剂B0.25-0.45%,黄原胶0.2-0.4%,丙烯酸树脂PemulenTR-20.3%和香料0.05%,所述百分比为相对于除氨甲基丙醇以外的各组分总重量的重量百分比;所述氨甲基丙醇的含量以调节所述皮肤外用剂的pH值至5.5-7为准;其中,所述含石榴提取物的组合物中,所述石榴提取物的含量为0.1-3.7wt%,所述冰川水的含量为96.3-99.9wt%;所述的多元醇类B为丁二醇和/或1,3-丙二醇;所述其他活性成分B为烟酰胺、抗坏血酸磷酸酯钠、生育酚乙酸酯和透明质酸钠中的一种或多种;所述表面活性剂B为羟乙基纤维素、异硬脂酸异丙酯、聚甘油-2油酸酯和聚甘油-10油酸酯中的一种或多种;所述防腐剂B为羟苯丙酯、羟苯甲酯和苯氧乙醇中的一种或多种。
按照本领域常识,丙烯酸树脂PemulenTR-2为一种丙烯酸(酯)类/C10-30烷醇丙烯酸酯交联聚合物,由美国诺誉化工公司生产。
在本发明的较佳实施方式三中,所述的皮肤外用剂为乳液,所述皮肤外用剂包含三乙醇胺,以及下述重量百分比的组分:所述含石榴提取物的组合物70-76.1%,甘油5%,1,3-丙二醇5%,泛醇1%,生育酚乙酸酯0.5%,其他活性成分C2-7.1%,聚二甲基硅氧烷2%,乳化剂A1652%,异硬脂酸异丙酯3%,油类C2-3%,鲸蜡硬脂醇0.5%,防腐剂C0.3%,黄原胶0.5%和EDTA二钠0.1%,所述百分比为相对于除三乙醇胺以外的各组分总重量的重量百分比;所述三乙醇胺的含量以调节所述皮肤外用剂的pH值至5.5-7为准;其中,所述含石榴提取物的组合物中,所述石榴提取物的含量为0.1-4.3wt%,所述冰川水的含量为95.7-99.9wt%;所述其他活性成分C为甘草酸二钾、烟酰胺和氢化聚异丁烯中的一种或多种;所述油类C为矿油和/或异十六烷;所述防腐剂C为羟苯甲酯和/或羟苯丙酯。
在本发明的较佳实施方式四中,所述的皮肤外用剂为乳霜,所述皮肤外用剂包含三乙醇胺,以及下述重量百分比的组分:所述含石榴提取物的组合物65.1-73.1%,甘油5%,1,3-丙二醇5%,泛醇1%,生育酚乙酸酯0.5%,其他活性成分D2-7.1%,聚二甲基硅氧烷2%,乳化剂A1652%,异硬脂酸异丙酯3%,油类D2-8%,鲸蜡硬脂醇2-3%,防腐剂D0.3%,丙烯酸羟乙酯/丙烯酰二甲基牛磺酸钠共聚物2%和EDTA二钠0.1%,所述百分比为相对于除三乙醇胺以外的各组分总重量的重量百分比;所述三乙醇胺的含量以调节所述皮肤外用剂的pH值至5.5-7为准;其中,所述含石榴提取物的组合物中,所述石榴提取物的含量为0.1-4.5wt%,所述冰川水的含量为95.5-99.9wt%;所述其他活性成分D为甘草酸二钾、烟酰胺和氢化聚异丁烯中的一种或多种;所述油类D为矿油和/或异十六烷;所述防腐剂D为羟苯甲酯和/或羟苯丙酯。
按照本领域常识,乳化剂A165为市售商品,其含有甘油硬脂酸酯和PEG-100硬脂酸酯。所述丙烯酸羟乙酯/丙烯酰二甲基牛磺酸钠共聚物为赛比克公司生产的牌号为SepinovEMT10的增稠剂。
在本发明的皮肤外用剂中,较佳地,水的来源仅为所述的冰川水,即是说,所述的皮肤外用剂中不含有其他类型的水,例如超纯水或自来水。
在本发明中,所述的室温为本领域常规意义上的室温,一般指20-30℃。
在符合本领域常识的基础上,上述各优选条件,可任意组合,即得本发明各较佳实例。
本发明所用试剂和原料均市售可得。
本发明的积极进步效果在于:
本发明的含石榴提取物的组合物能够显著地抑制HEM-m细胞黑色素的生成,并且天然、安全,可用作皮肤外用剂的美白活性成分。本发明的皮肤外用剂成分天然、安全,并具有美白的功效。
附图说明
图1为实施例3中不同浓度的石榴提取物对HEM-m黑色素生成的影响的OD490柱状图。
图2为实施例4中各样品对HEM-m黑色素生成的影响的OD470柱状图。
具体实施方式
下面通过实施例的方式进一步说明本发明,但并不因此将本发明限制在所述的实施例范围之中。下列实施例中未注明具体条件的实验方法,按照常规方法和条件,或按照商品说明书选择。
下述实施例中,所使用的冰川水取自喜马拉雅山的多吉曲登尼玛泉(海拔5128米),每升的该冰川水含有下述组分:Sr+0.2-0.5mg、K+0.5-10.0mg、Na+1.0-5.0mg、Ca2+14.0-30.0mg以及Mg2+2.0-10.0mg;溶解性总固体含量为60-150mg/L;冰川水的pH值为7.4,该冰川水中的氘含量为133ppm。
下述实施例中,所使用的石榴提取物由下述制备方法制得:
取云南玉溪产酸石榴,分离其皮、籽部位,石榴皮粉碎至粗粉,所述粗粉的粒径为40-100目,加8-12倍体积50-90%乙醇超声提取2次,每次2小时,合并滤液,浓缩至150mL,经AB-8大孔吸附树脂富集,依次以水、30%乙醇和70%乙醇洗脱,收集70%乙醇洗脱部位,浓缩至干,得石榴提取物。
实施例1DMEM(H)培养基的配制及HEM-m细胞的培养
实验材料
超纯水(CK)、冰川水、DMEM高糖培养基(培养基干粉购自GIBCO公司,NO.12800-017)、胎牛血清(购自GOBCO公司,NO.10099-141)、胰酶消化液(Sigma公司)、人源表皮黑色素细胞HEM-m(取自于上海市瑞金医院泌尿科切除的人体包皮)。
实验方法
(1)分别用超纯水(CK)、冰川水按照培养基干粉说明书配制DMEM高糖培养基0.22μm无菌过滤,分别加入胎牛血清配制成含10%胎牛血清的DMEM(H)培养基。
(2)等培养瓶中的细胞HEM-m长至80%融合状态时,倒去原来的培养液,用PBS洗涤,之后加入胰酶细胞消化液进行消化。胰酶消化过程可以将培养瓶放置在37℃培养箱中。
(3)待贴壁的HEM-m细胞漂浮在液体之中并且相互散开之时,加入2mL胎牛血清终止消化。
(4)吹打并收集细胞,1000r/min离心3分钟,弃去上清液,加入PBS重新悬浮细胞,使细胞呈相互分离状态。
(5)用含10%胎牛血清的超纯水配制的DMEM(H)培养基进行传代。
(6)将传代好的细胞置于37℃,5%CO2恒温培养箱中进行培养,获得传代的HEM-m细胞。
实施例2人表皮黑色素细胞HEM-m增殖活性的测定
采用MTT法,待细胞培养瓶中HEM-m细胞长至80%融合度时,胰酶消化并细胞计数,吸取细胞,并调整细胞浓度为5×104个/mL,充分混匀,取100μl接入96孔板的孔中。每组设置6个复孔,37℃孵育72小时后。将5×MTT用稀释液稀释为1×MTT。每孔加入50μL1×MTT,在37℃孵育4小时。吸出上清液,每孔加入150μL二甲基亚砜(DMSO),放置在平板摇床上10分钟充分摇匀,摇匀后用酶标仪在490nm处检测每孔的光密度(OD490)。采用超纯水DMEM(H)和石榴提取物的培养基浓度梯度设置如下:0.00%、0.02%、0.015%、0.01%、0.008%、0.005%、0.002%、0.001%。
实验结果如图1所示。从图中可以看出,当石榴提取物终浓度为0.005%时,其对HEM-m的增殖活性无显著性差异(p=0.925>0.05),即认为此浓度不会对HEM-m的增殖产生影响。因此将0.005%作为检测黑色素含量时石榴的终浓度。
实施例3人表皮黑色素细胞HEM-m黑色素含量测定
采用改良的HideyaAndo法,选择对数生长期的选择对数生长期的B-16黑素瘤细胞,以2×104个/mL的密度接种于6孔培养板,每孔2.5mL,置于37℃,5%CO2饱和湿度环境中进行培养。分别用超纯水Ⅱ、冰川水配制的含10%胎牛血清的DMEM(H)培养基,实验时,加入石榴提取物,得到实验用的4种培养基:
超纯水DMEM(H),记为CK组;
冰川水DMEM(H),记为冰川水组;
超纯水DMEM(H)+石榴提取物,记为CK+石榴组;
冰川水DMEM(H)+石榴提取物,记为冰川水+石榴组。
石榴提取物终浓度参考MTT实验的结果得到为0.005%。分别用4种培养基培养HEM-M细胞72小时后,分别收集细胞,调整细胞浓度到3×105个/mL,吸取1mL细胞悬液分别置于离心管中,离心去上清,用200μLPBS缓冲液重悬细胞沉淀后,加入1mL1:1乙醇乙醚液,室温下放置30分钟,3000r/min离心5分钟后弃上清。加入1mL含10%二甲基亚砜(DMSO)的1mol/LNaOH溶液,80℃水浴45分钟后于470nm波长处测定吸光度值(OD470)。每组设三个平行对照。
测试结果如图2所示。从图中可以看出,和对照组(CK)比较,将冰川水和石榴提取物相配合,能显著抑制黑色素生成。各样品的对人表皮黑色素细胞HEM-m黑色素的抑制作用大小排序为:冰川水+石榴>CK+石榴≈冰川水>CK;具体的黑色素抑制率结果见下表1。
表1各组分对人表皮黑色素细胞HEM-m黑色素的抑制作用
样品 | 黑色素抑制率(%) |
CK | 0 |
冰川水 | 9.25 |
CK+石榴 | 9.23 |
冰川水+石榴 | 15.03 |
实施例4-6皮肤外用剂
实施例4-6的皮肤外用剂为化妆水,其具体组成见下表2。
表2
按照本领域常规的化妆水的制备方法,制得实施例4-6的皮肤外用剂。
实施例7-9皮肤外用剂
实施例7-9的皮肤外用剂为精华液,其具体组成见下表3。
表3
表3中,百分比均为相对于除氨甲基丙醇以外的各组分总重量的重量百分比。按照本领域常规的精华液的制备方法,制得实施例7-9的皮肤外用剂。
实施例10-15皮肤外用剂
实施例10-12的皮肤外用剂为乳液,实施例13-15的皮肤外用剂为乳霜,其具体组成分别见下表4。
表4
表4中,百分比均为相对于除三乙醇胺以外的各组分总重量的重量百分比。按照本领域常规的乳液的制备方法,制得实施例10-12的皮肤外用剂;按照本领域常规的乳霜的制备方法,制得实施例13-15的皮肤外用剂。
以上所述仅为本发明的较佳实施例而已,并不用以限制本发明,凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等,均应包含在本发明的保护范围之内。
Claims (11)
1.一种含石榴提取物的组合物,其特征在于,其包括下述质量百分比的组分:石榴提取物0.001-4.7%和冰川水95.3-99.999%。
2.如权利要求1所述的含石榴提取物的组合物,其特征在于,所述的石榴提取物由下述制备方法制得:取酸石榴,分离其皮、籽部位,粉碎至粗粉,所述粗粉的粒径为40-100目,加8-12倍体积50-90%乙醇超声提取2-3次,每次1-3小时,合并滤液,浓缩,经AB-8大孔吸附树脂富集,依次以水、30%乙醇和70%乙醇洗脱,收集70%乙醇洗脱部位,浓缩至干,得石榴提取物。
3.如权利要求1或2所述的含石榴提取物的组合物,其特征在于,所述冰川水的pH值为7.1-7.9,所述冰川水中的氘含量为133-137ppm,且所述冰川水中不含溴酸盐。
4.如权利要求3所述的含石榴提取物的组合物,其特征在于,每升所述冰川水含有下述组分:Sr+0.2-0.5mg、K+0.5-10.0mg、Na+1.0-5.0mg、Ca2+14.0-30.0mg以及Mg2+2.0-10.0mg;所述冰川水中的溶解性总固体含量为60-150mg/L。
5.如权利要求1-4中任一项所述含石榴提取物的组合物的用途,其特征在于,所述含石榴提取物的组合物作为皮肤外用剂中的美白活性成分和/或保湿活性成分。
6.如权利要求5所述含石榴提取物的组合物的用途,其特征在于,所述美白活性成分为抑制人源表皮黑色素细胞HEM-m的黑色素生成的美白活性成分。
7.一种皮肤外用剂,其特征在于,其包括如权利要求1-4中任一项所述的含石榴提取物的组合物。
8.如权利要求7所述的皮肤外用剂,其特征在于,所述含石榴提取物的组合物在所述皮肤外用剂中的含量为65.1-89.2wt%。
9.如权利要求7或8所述的皮肤外用剂,其特征在于,所述的皮肤外用剂包含其他活性成分,所述其他活性成分为保湿活性成分、抗氧化活性成分、控油活性成分、美白活性成分和抗衰老活性成分中的一种或多种;
和/或,所述的皮肤外用剂含有赋形剂;所述赋形剂的形式为水相、油相、凝胶、水包蜡型乳液、水包油型乳液或油包水型乳液的形式;
和/或,所述的皮肤外用剂包含防腐剂、芳香剂以及亲水性和亲脂性活性剂中的一种或多种;
和/或,所述皮肤外用剂包含颗粒相,所述的颗粒相为颜料、珠光剂和填充剂中的一种或多种。
10.如权利要求9所述的皮肤外用剂,其特征在于,当所述的皮肤外用剂为化妆水时,所述的皮肤外用剂包含下述重量百分比的组分:所述含石榴提取物的组合物84.2-89.2%,甘油5%,多元醇类A3-6%,甜菜碱1%,泛醇0.5%,甘草酸二钾0.05-0.1%,其他活性成分A0.11-2.0%,聚乙二醇-320.5%,表面活性剂A0.15-0.2%,防腐剂A0.4-0.45%,黄原胶0-0.15%和香料0.03%;其中,所述含石榴提取物的组合物中,所述石榴提取物的含量为0.001-3.6wt%,所述冰川水的含量为96.4-99.999wt%;所述的多元醇类A为丁二醇、1,2-戊二醇和双丙甘醇中的一种或多种;所述其他活性成分A为尿囊素、烟酰胺和透明质酸钠中的一种或多种;所述表面活性剂A为羟乙基纤维素、聚山梨醇酯-20和PEG-40氢化蓖麻油中的一种或多种;所述防腐剂A为防腐剂IS-45、羟苯甲酯和苯氧乙醇中的一种或多种;
当所述的皮肤外用剂为精华液时,所述皮肤外用剂包含氨甲基丙醇,以及下述重量百分比的组分:所述含石榴提取物的组合物82.4-85.6%,甘油5%,多元醇类B3-6%,泛醇1%,甘草酸二钾0.1%,尿囊素0.1%,其他活性成分B2.1-4.3%,表面活性剂B0.05-3.6%,防腐剂B0.25-0.45%,黄原胶0.2-0.4%,丙烯酸树脂PemulenTR-20.3%和香料0.05%,所述百分比为相对于除氨甲基丙醇以外的各组分总重量的重量百分比;所述氨甲基丙醇的含量以调节所述皮肤外用剂的pH值至5.5-7为准;其中,所述含石榴提取物的组合物中,所述石榴提取物的含量为0.004-3.7wt%,所述冰川水的含量为96.3-99.996wt%;所述的多元醇类B为丁二醇和/或1,3-丙二醇;所述其他活性成分B为烟酰胺、抗坏血酸磷酸酯钠、生育酚乙酸酯和透明质酸钠中的一种或多种;所述表面活性剂B为羟乙基纤维素、异硬脂酸异丙酯、聚甘油-2油酸酯和聚甘油-10油酸酯中的一种或多种;所述防腐剂B为羟苯丙酯、羟苯甲酯和苯氧乙醇中的一种或多种;
当所述的皮肤外用剂为乳液时,所述皮肤外用剂包含三乙醇胺,以及下述重量百分比的组分:所述含石榴提取物的组合物70-76.1%,甘油5%,1,3-丙二醇5%,泛醇1%,生育酚乙酸酯0.5%,其他活性成分C2-7.1%,聚二甲基硅氧烷2%,乳化剂A1652%,异硬脂酸异丙酯3%,油类C2-3%,鲸蜡硬脂醇0.5%,防腐剂C0.3%,黄原胶0.5%和EDTA二钠0.1%,所述百分比为相对于除三乙醇胺以外的各组分总重量的重量百分比;所述三乙醇胺的含量以调节所述皮肤外用剂的pH值至5.5-7为准;其中,所述含石榴提取物的组合物中,所述石榴提取物的含量为0.13-4.3wt%,所述冰川水的含量为95.7-99.87wt%;所述其他活性成分C为甘草酸二钾、烟酰胺和氢化聚异丁烯中的一种或多种;所述油类C为矿油和/或异十六烷;所述防腐剂C为羟苯甲酯和/或羟苯丙酯;
当所述的皮肤外用剂为乳霜时,所述皮肤外用剂包含三乙醇胺,以及下述重量百分比的组分:所述含石榴提取物的组合物65.1-73.1%,甘油5%,1,3-丙二醇5%,泛醇1%,生育酚乙酸酯0.5%,其他活性成分D2-7.1%,聚二甲基硅氧烷2%,乳化剂A1652%,异硬脂酸异丙酯3%,油类D2-8%,鲸蜡硬脂醇2-3%,防腐剂D0.3%,丙烯酸羟乙酯/丙烯酰二甲基牛磺酸钠共聚物2%和EDTA二钠0.1%,所述百分比为相对于除三乙醇胺以外的各组分总重量的重量百分比;所述三乙醇胺的含量以调节所述皮肤外用剂的pH值至5.5-7为准;其中,所述含石榴提取物的组合物中,所述石榴提取物的含量为0.15-4.5wt%,所述冰川水的含量为95.5-99.85wt%;所述其他活性成分D为甘草酸二钾、烟酰胺和氢化聚异丁烯中的一种或多种;所述油类D为矿油和/或异十六烷;所述防腐剂D为羟苯甲酯和/或羟苯丙酯。
11.如权利要求7-10任一项所述的皮肤外用剂,其特征在于,所述的皮肤外用剂中,水的来源仅为所述的冰川水。
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