CN108653064B - 雪茶提取物及其制备方法和应用 - Google Patents
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Abstract
本发明属于植物提取领域及日化领域,具体涉及一种雪茶提取物,制备方法为:取干燥的雪茶叶片粉末,加入冰川水,超声提取后取滤液,浓缩或者冷冻干燥。用上述提取方法,提取效果有显著提高,所获得的雪茶提取物能够明显促进皮肤成纤维细胞的增殖,有效提高I型胶原蛋白的合成,并且提高成纤维细胞表达透明质酸,可作为活性成分用于制备皮肤外用剂,有利于皮肤维持较高含水量,保持皮肤湿润,减少皱纹的产生;并且能够有效促进成纤维细胞分泌胶原蛋白、弹性纤维蛋白及多种细胞修复因子,从而修复老化的皮肤和延缓皮肤老化,具有抗衰老的作用,具有保湿、抗衰老或修复功能。
Description
技术领域
本发明属于植物提取领域及日化领域,具体涉及一种雪茶提取物及其应用。
背景技术
雪茶是滇西北少数民族传统的保健茶,主要分布于海拔4000米左右的雪域高原地带,是雪域高原上天然的保健饮品和药物,分为白雪茶和红雪茶。白雪茶[Thamnoliavermicularis(Sw.)Ach]是地茶科地茶属雪茶和雪地茶的干燥叶状体。白雪茶形似白菊花瓣,茶汤清爽,风味独特,性凉味甘;含丰富的Ca、Fe元素,可用于治疗中暑、心中烦热、咳嗽、神经衰弱和高血压等症。
红雪茶[Lethariella cladonioides]具有消炎、镇静、止痛的功能,可治疗神经衰弱、头目眩晕。
多项研究报道雪茶中已分离出缩酚酸类、甾体类、多糖类等上百个化合物,且其丰富的生物学活性已得到验证。如雪茶素具有抗炎和免疫调节功能,羊角衣酸有较强的抗肿瘤活性,松萝酸则具有一定的抗菌活性。现有的报道中,用醇溶剂提取白雪茶,得到的白雪茶提取物中含有鳞片衣酸1和羊角衣酸2,能够用于治疗阿尔兹海默症。
研究表明,雪茶的水提取物具有抗疲劳、抗辐射、抗缺氧、降脂保肝等效果,但是雪茶活性成分在延缓皮肤衰老方面的作用至今尚无报道。
科学研究表明,氘对生命体的生存发展和繁衍是有害的,而自然界中的冰川水具有天然的低氘特性,可改善机体基础代谢水平、延缓衰老,被科学界称之为“生命之水”。冰川水多用来作为活性成分添加到护肤品等皮肤外用剂中,增强镇静皮肤、补水保湿、抗氧化、抗衰老等效果。目前也没有使用冰川水作为媒介提取植物活性成分的报道。
发明内容
本发明旨在提供一种雪茶提取物及制备方法。
本发明还将上述雪茶提取物应用于皮肤外用剂,尤其是具有保湿、抗衰老和修复皮肤功能的化妆品。
技术方案为:一种雪茶提取物的制备方法,包括以下步骤:取干燥的雪茶叶片粉末,加入冰川水,超声提取后取滤液。可将滤液进一步浓缩或者冷冻干燥。
优选的,制备方法包括以下步骤:取干燥的雪茶叶片粉末,加入冰川水,超声提取1~3次,合并滤液;每次提取的冰川水与雪茶叶片粉末用量比为5~30L:1kg,优选为10~25L:1kg;每次超声提取的时间为10~45min,功率300~1500W。
可将滤液进一步浓缩或者冷冻干燥。优选的,在-5~-20℃下冷冻干燥,更优选为-10~-15℃。
所述的雪茶叶片粉末含水量低于5%,粒径为50~200目。雪茶叶片为红雪茶或白雪茶叶片。
本发明的一个优选方案为:取50~200目干燥的雪茶叶片粉末,加入冰川水,超声提取2次,合并滤液,浓缩或者冷冻干燥;每次提取的冰川水与雪茶叶片粉末用量比为15~25L:1kg,每次超声提取的时间为20~40min,功率300~800W。
优选的,所述的冰川水含有如下组分:Sr2+0.2~0.5mg/L、K+0.5~10.0mg/L、Na+1.0~5.0mg/L、Ca2+14.0~30.0mg/L以及Mg2+2.0~10.0mg/L;并且冰川水中不含溴酸盐,pH值为7.1~7.9,氘含量为130~140ppm,不含有不溶性固体物或者悬浮物,溶解性总固体含量为60~150mg/L。优选的,pH值为7.2~7.6,更优选为7.4。
通过上述方法所获得的雪茶提取物,能够促进皮肤成纤维细胞的增殖和表达透明质酸,增加I型胶原蛋白的合成,可用于制备皮肤外用剂,例如化妆品。以干物质计,雪茶提取物在皮肤外用剂中的用量为0.001wt%~20wt%,优选为0.01wt%~5wt%。更优选的,所述皮肤外用剂中还含有冰川水。
所述的皮肤外用剂为具有保湿、抗衰老或修复功能的皮肤外用剂,例如化妆品,类型为洁面霜、乳液、霜膏、精华液、化妆水(如柔肤水、爽肤水)、面膜、卸妆水等。所述的皮肤外用剂如化妆品能够促进皮肤成纤维细胞增殖、I型胶原蛋白合成以及促进皮肤成纤维细胞表达透明质酸,具有保湿、防皱功能和抗衰老功能,能够延缓皮肤衰老和修复老化的皮肤。
优选的,所述的皮肤外用剂中还含有冰川水。这种皮肤外用剂具有更好的抗衰老作用。
人皮肤成纤维细胞是皮肤真皮网织层中最重要的细胞,是皮肤衰老和细胞受损后的主要修复细胞之一。它不但能够促进表皮细胞的迁移、增殖和分化,还能分泌大量的胶原蛋白、弹性纤维蛋白及多种细胞修复因子,具有强大的自我更新能力,从而修复老化的皮肤。一般来说,真皮衰老表现为真皮对外来化学物清除力下降,真皮厚度变薄、胶原蛋白和弹性蛋白合成减少、分解增加,分解酶活性增强。这些现象都与成纤维细胞数减少以及分泌合成功能下降或异常有关。如果能够提高皮肤成纤维细胞的增殖水平,促进I型胶原蛋白的合成率,提高成纤维细胞的透明质酸表达,则可以延缓皮肤衰老,修复皮肤的功能并实现保湿的效果。
本发明采用冰川水提取雪茶活性成分,与使用纯水相比,提取效果有显著提高,收率可提高30%以上。提取过程中不使用有机溶剂,也不需要加热,提取方法简单,对环境友好,能够更好地保留雪茶中的活性成分。所获得的雪茶提取物能够明显促进皮肤成纤维细胞的增殖,有效提高I型胶原蛋白的合成,并且提高成纤维细胞表达透明质酸,具有很好的保湿、抗衰老作用。
采用这种雪茶提取物作为活性成分的皮肤外用剂,能够显著提高成纤维细胞透明质酸含量,有利于皮肤维持较高含水量,保持皮肤湿润,减少皱纹的产生;并且能够有效促进成纤维细胞分泌胶原蛋白、弹性纤维蛋白及多种细胞修复因子,从而修复老化的皮肤和延缓皮肤老化,具有抗衰老的作用。
雪茶提取物与冰川水共同用于制备皮肤外用剂如化妆品时,抗衰老的效果更为明显。
具体实施方式
实施例1
取白雪茶叶片(含水量低于5%)粉碎后过80目筛,取10g粉末加入200mL的冰川水,超声(500W)30min;离心过滤,收集滤液。
再加入200mL冰川水重复提取1次,滤液合并后,在-10~-15℃下冷冻干燥,收集固体,得到雪茶提取物。
冰川水取自喜马拉雅山的多吉曲登尼玛泉(海拔5128米),冰川水中的氘含量为134ppm,pH=7.4,不含溴酸盐,含有以下的组分:Sr2+0.2~0.5mg/L、K+0.5~10.0mg/L、Na+1.0~5.0mg/L、Ca2+14.0~30.0mg/L以及Mg2+2.0~10.0mg/L;并且冰川水不含有不溶性固体物或者悬浮物,其中的溶解性总固体含量为80~100mg/L。
用超纯水代替冰川水,按同样方法进行提取。两种方法的提取结果如表1。以原料重量计,用超纯水提取后得到的雪茶提取物3g,收率为3%;用冰川水提取获得的雪茶提取物为4g,收率4%。结果显示,采用冰川水提取,可以提高雪茶的提取率。
400mL冰川水中的溶解性总固体约为0.04g,由此可见用冰川水得到的雪茶提取物重量增加的原因不在于冰川水本身所含的干物质,而是是采用冰川水后,对于雪茶活性成分的提取效果显著提高。
表1超纯水和冰川水提取白雪茶的提取率
得率(%) | |
超纯水提取 | 3 |
冰川水提取 | 4 |
实施例2雪茶提取物促进成纤维细胞增殖活性的检测
一般来说,真皮衰老表现为真皮对外来化学物清除力下降,真皮厚度变薄、胶原蛋白和弹性蛋白合成减少、分解增加,分解酶活性增强。这些现象都与成纤维细胞数减少以及分泌合成功能下降或异常有关。人皮肤成纤维细胞是皮肤真皮网织层中最重要的细胞,是皮肤衰老和细胞受损后的主要修复细胞之一。它不但能够促进表皮细胞的迁移、增殖和分化,还能分泌大量的胶原蛋白、弹性纤维蛋白及多种细胞修复因子,具有强大的自我更新能力,从而修复老化的皮肤。如果活性成分能够促进成纤维细胞的增殖或合成I型胶原蛋白,说明该活性成分具有抗衰老作用。
以超纯水和冰川水分别配制含10%胎牛血清的DMEM培养基培养人成纤维细胞。将实施例1的雪茶超纯水提取物和雪茶冰川水提取物分别加入超纯水配制的细胞培养基中,含量为0.1mg/mL;冰川水配制的细胞培养基中加入实施例1的雪茶冰川水提取物,含量为0.1mg/mL。
培养24小时后,用MTT法对细胞染色,用酶标仪测定550nm处吸光度。以空白对照的增殖率为100%,参比空白对照,评估对人成纤维细胞的增殖作用;同时取细胞上清样本,用I型胶原蛋白测定试剂盒测定胶原蛋白的生成,空白对照的生成率为100%,参比空白对照,评估对人成纤维细胞的I型胶原合成促进作用。
由结果可知,与空白对照组和雪茶超纯水提取物相比相比,用冰川水或者雪茶冰川水提取物处理后,成纤维细胞的增殖活性和I型胶原合成都有显著提高;冰川水与雪茶冰川水提取物两者共同作用时,促进成纤维细胞增殖和I型胶原合成的效果更加显著。
表2不同处理对成纤维细胞增殖以及I型胶原合成的影响
实施例3 ELISA法检测人成纤维细胞透明质酸的表达
透明质酸也是细胞外基质的重要成分,有维持皮肤含水量、调节生理水平衡、促进营养物质的运输和细胞代谢的重要功能。皮肤中的透明质酸随年龄增加而逐渐流失,因此,在皮肤衰老过程中提高透明质酸含量有利于维持皮肤较高的含水量,保持皮肤湿润而减少皱纹的发生。如果活性物质能够促进透明质酸表达,增加细胞外基质中的透明质酸含量,说明具有良好的保湿效果。
按实施例2的方法,用超纯水和冰川水分别配制含10%胎牛血清的DMEM培养基培养人成纤维细胞,雪茶提取物加入到细胞培养基中。培养24小时后,收集细胞并用PBS清洗,离心收集上清备用。按照透明质酸ELISA试剂盒说明书方法进行操作,用酶标仪测定样品450nm下吸光值,根据绘制的标准曲线计算出细胞透明质酸浓度。结果显示,与空白对照和雪茶超纯水提取物相比,冰川水组、雪茶冰川水提取物、以及冰川水和雪茶冰川水提取物共同处理后,细胞外基质中的透明质酸含量显著提高。
表3不同处理对成纤维细胞透明质酸含量的影响(ng/mL)
透明质酸含量 | |
空白对照 | 26.43 |
冰川水组 | 27.49 |
雪茶超纯水提取物(0.1mg/mL) | 26.78 |
雪茶冰川水提取物(0.1mg/mL) | 32.06 |
雪茶冰川水提取物(0.1mg/mL)+冰川水 | 32.19 |
用红雪茶作为原料代替实施例1中的白雪茶,采用冰川水和超纯水用相同的方法提取,所获得的雪茶冰川水提取物收率同样高于雪茶超纯水提取物,提高约25%。与空白对照组、超纯水提取物相比,雪茶冰川水提取物以及雪茶冰川水提取物与冰川水的混合液能够明显促进成纤维细胞增殖以及I型胶原蛋白合成。在超纯水或冰川水配制含10%胎牛血清的DMEM培养基中加入上述雪茶冰川水提取物(0.1mg/mL),与空白对照组相比,培养的人成纤维细胞透明质酸表达均有显著提高。
Claims (8)
1.一种雪茶提取物的制备方法,其特征在于,包括以下步骤:取干燥的雪茶叶片粉末,加入冰川水,超声提取后取滤液;
所述的冰川水不含溴酸盐,pH值为7.1~7.9,氘含量为130~140ppm,还含有如下组分:Sr2+ 0.2~0.5mg/L、K+ 0.5~10.0mg/L、Na+ 1.0~5.0mg/L、Ca2+ 14.0~30.0mg/L以及Mg2+2.0~10.0mg/L;并且冰川水中的溶解性总固体含量为60~150mg/L;所述的雪茶叶片为白雪茶叶片。
2.根据权利要求1所述雪茶提取物的制备方法,其特征在于,制备方法包括以下步骤:
取干燥的雪茶叶片粉末,加入冰川水,超声提取1~3次,合并滤液;每次提取的冰川水与雪茶叶片粉末用量比为5~30L:1kg,每次超声提取的时间为10~45min。
3.根据权利要求1或2所述雪茶提取物的制备方法,其特征在于,所述的雪茶叶片粉末含水量低于5%,粒径为50~200目。
4.根据权利要求1或2所述雪茶提取物的制备方法,其特征在于,将滤液浓缩或者冷冻干燥。
5.一种雪茶提取物,其特征在于,制备方法通过权利要求1~4任一项所述的方法制备。
6.权利要求5所述雪茶提取物在制备具有保湿、抗衰老或修复功能皮肤外用剂方面的应用。
7.权利要求6所述的应用,其特征在于,以干物质计,雪茶提取物在皮肤外用剂中的含量为0.001wt%~5wt%。
8.权利要求6或7所述的应用,其特征在于,所述皮肤外用剂中还含有冰川水。
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