CN117414322A - 一种含有紧致抗皱组合物的精华液及其制备方法 - Google Patents
一种含有紧致抗皱组合物的精华液及其制备方法 Download PDFInfo
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- CN117414322A CN117414322A CN202311489173.XA CN202311489173A CN117414322A CN 117414322 A CN117414322 A CN 117414322A CN 202311489173 A CN202311489173 A CN 202311489173A CN 117414322 A CN117414322 A CN 117414322A
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Abstract
本发明公开了一种含有紧致抗皱组合物的精华液及其制备方法,属于化妆用的配制品领域;所述含有紧致抗皱组合物的精华液包括的组分有:紧致抗皱组合物、棕榈酰三肽‑5、羟丙基四氢吡喃三醇,以及溶剂;其中紧致抗皱组合物包括黑孢块菌提取物、芍药根提取物、缬草根提取物、青钱柳提取物。本发明提供的含有紧致抗皱组合物的精华液对弹性蛋白酶活性的抑制率达到了65%以上,对I型胶原蛋白合成具有显著的促进作用,同时,对自由基的清除也具有优异的效果。
Description
技术领域
本发明属于化妆用的配制品领域,具体涉及一种含有紧致抗皱组合物的精华液及其制备方法。
背景技术
随着年龄的增长,皮肤会出现衰老的情况,而皮肤最初的衰老是从眼周开始的。由于眼周肌肤比面部肌肤更加脆弱,眼周肌肤水分蒸发速度较快,同时,眼周皮肤的汗腺和皮脂腺分布较少,使得眼部皮肤里面的胶原蛋白飞速流失,眼部皮肤更容易衰老,眼袋、泪沟、鱼尾纹都是眼睛衰老发出的信号。
弹性蛋白位于皮肤真皮层,存在真皮层的细胞间隙之间,与胶原蛋白等其他细胞外基质一起维持组织结构的稳定性,主要提供细胞间的黏附和支持作用,同时其很强的伸缩性和弹性也是维系皮肤弹性的重要原因。弹性蛋白最大的特点就是结构和性质非常非常稳定。既不容易被溶解也不容易被降解,而且还具有弹性,可拉伸数倍于原来的长度。而皮肤衰老大体可以分为自然老化和光老化。人在自然老化过程中,弹力纤维会被降解。弹力纤维虽然特别稳定,但是人体中依旧有种酶可以降解它,叫弹性蛋白酶。弹性蛋白酶是基质金属蛋白酶家族中最重要的一种酶之一,主要由成纤维细胞合成分泌,能够降解皮肤中弹性蛋白导致皮肤老化。
随着年龄的增长,人体对弹性蛋白酶的抑制能力越来越弱,同时弹性蛋白酶的活性越来越强,弹性蛋白酶不仅能够降解包括弹性蛋白在内的大部分胞外基质蛋白,还能激活其他基质金属蛋白酶活性,加速真皮层细胞外基质的降解,从而导致人类皮肤真皮层正常的结构和功能被严重损害,使皮肤产生明显的皱纹,失去弹性,因此抑制弹性蛋白酶的活性在一定程度上能起到紧致抗皱的功效。
目前常规的紧致抗皱化妆品,其效果总是不尽人意,原料间组合的协同效果差,不仅不能起到增效的作用,甚至可能彼此消弭,弱化各组分单独作用时功效,因此开发出一种具有紧致抗皱功效的产品具有重要意义。
发明内容
针对现有技术的不足,本发明目的在于提供一种有效抑制弹性蛋白酶活性且原料组分间协同作用优异的紧致抗皱组合物以及含有该组合物的精华液和制备方法。
为实现上述目的,本发明公开了一种含有紧致抗皱组合物的精华液,包括溶剂以及以下质量份数的组分:
紧致抗皱组合物 1.5-2份;
棕榈酰三肽-5 0.3-1份;
羟丙基四氢吡喃三醇 1.4-2.2份;
所述紧致抗皱组合物包括黑孢块菌提取物、芍药根提取物、缬草根提取物、青钱柳提取物。
具体地,所述黑孢块菌提取物、芍药根提取物、缬草根提取物和青钱柳提取物的质量比为1:(0.23-0.57):(5.45-6.70):(2.64-3.11)。
黑孢块菌提取物含有种类丰富的多糖、氨基酸、多肽、维生素等成分,具有显著的抗氧化、清除自由基的作用,同时还能加强保湿、修复损伤的皮肤,恢复皮肤的屏障和正常代谢功能,其中的多糖成分可帮助清除机体中过多的DPPH自由基和羧基自由基,以抵消自由基对人体细胞的氧化攻击,修复其对皮肤的氧化侵蚀损伤,减缓皮肤的衰老进程,减少皱纹,暗黄等皮肤问题。
芍药根提取物含有丰富的活性组分,包括芍药苷、黄酮类化合物、三萜化合物、酚酸化合物、活性多糖、活性肽等,对于弹性蛋白酶、透明质酸酶、酪氨酸酶均有很好的抑制效果,抗氧化效果佳,对于紫外线损伤的细胞修复有很好的作用。
缬草根提取物富含多种挥发性成分,如α、β-蒎烯、缬草萜酮、缬草萘烯醇、缬草萜烯醇酸、缬草萜烯酸等,具有皮肤美白、色素沉着抑制、减少UV-吸收性等作用,其还具有减少皮脂生成,抵抗皮脂过量生成,调节皮肤和毛囊失调的功效。
青钱柳提取物含有丰富的抗氧化活性成分,包括黄酮类化合物、多糖类化合物、萜类化合物、无机元素及其维生素E、维生素C等抗氧化物质,能够有效与自由基结合,起到清除自由基的作用,延迟皮肤衰老。
通过本发明研究发现,黑孢块菌提取物、芍药根提取物、缬草根提取物和青钱柳提取物之间具有共同促进皮肤修复的作用,协同作用下能够有效提高抑制弹性蛋白酶活性的能力,提高促进皮肤胶原蛋白的合成的作用,增强对自由基离子的清除作用。
具体地,所述缬草根提取物制备方法包括以下步骤:
(1)取一定量缬草根切碎,加水搅拌均匀,然后添加复配微生态制剂,发酵24-48h后,得缬草根粗提物;
(2)将缬草根粗提物过滤去渣后浓缩,得缬草根提取物。
更具体地,所述步骤(1)中的缬草根与水的质量比为1:(1.0-1.3);所述复配微生态制剂的添加量为所述缬草根与水总质量的1-2%,复配微生态制剂中的微生物包括枯草芽孢杆菌和凝结芽孢杆菌,所述枯草芽孢杆菌的活菌数≥100亿CFU/g,凝结芽孢杆菌的活菌数≥50亿CFU/g;
所述步骤(2)中,缬草根粗提物通过≥60目的滤网过滤,然后浓缩至含水为10-15%的膏状物,得缬草根提取物。
缬草根通过复配微生态制剂发酵后能够有效的释放缬草根中的活性营养物质;复配微生态制剂在发酵中通过释放蛋白酶、脂肪酶、纤维素酶,来分解植物细胞壁,使得缬草根在普通的物理破碎过程中无法被释放出来的活性营养成分,能够被充分的释放并溶解到溶剂中,提高了缬草根提取物的提取率。
具体地,所述的含有紧致抗皱组合物的精华液还包括但不限于以下组分:螯合剂、增稠剂、保湿剂、pH调节剂、防腐剂。
更具体地,所述增稠剂为羟丙基甲基纤维素、黄原胶、叔丁醇中的至少一种。
更具体地,所述保湿剂为透明质酸钠、甜菜碱、甘油、丁二醇、赤藓醇中的至少一种。
更具体地,所述防腐剂为丙二醇、辛酰羟肟酸、1,2-己二醇、乙基己基甘油、对羟基苯乙酮中的至少一种。
更具体地,所述螯合剂为EDTA二钠、EDTA四钠中的至少一种;
所述pH调节剂为氨甲基丙醇、柠檬酸钠、氢氧化钠中的至少一种。
本发明还公开了前述的含有紧致抗皱组合物的精华液的制备方法,包括以下步骤:
步骤(1),将所述溶剂、螯合剂、增稠剂按一定量进行混合搅拌,得混合物;
步骤(2),将所述保湿剂、紧致抗皱组合物、棕榈酰三肽-5、羟丙基四氢吡喃三醇、pH调节剂、防腐剂按一定量加入混合物中,搅拌,得所述含有紧致抗皱组合物的精华液。
本发明的有益效果:
1.本发明提供的紧致抗皱组合物对弹性蛋白酶抑制率达到了65%以上,对I型胶原蛋白合成具有显著的促进作用,并对自由基的清除也具有显著的效果。
2.本发明提供的精华液能够修复损伤的皮肤,减缓皮肤的衰老进程,减少皱纹,暗黄等皮肤问题;对于弹性蛋白酶具有很好的抑制效果,抗氧化效果优异,对于紫外线损伤的细胞修复有很好的作用。
具体实施方式
为更好地说明本发明的目的、技术方案和优点,下面将结合具体实施例对本发明作进一步说明。本领域技术人员应当理解,此处所描述的具体实施例仅用以解释本发明,并不用于限定本发明。
实施例和对比例中所用的试验方法如无特殊说明,均为常规方法;所用的材料、试剂等,如无特殊说明,均可从商业途径得到;实施例和对比例中所提及的百分比,如无特殊说明,均为质量百分比。
本发明中:
黑孢块菌提取物为黑孢块菌(TUBER MELANOSPORUM)提取物,INCI名称为TUBERMELANOSPORUM EXTRACT,CAS No.:85085-76-3,购自西安尚诚生物科技有限公司。
芍药根提取物为芍药(PAEONIAALBIFLORA)根提取物,INCI名称为PAEONIAALBIFLORA ROOT EXTRACT,CAS No.:84929-40-8,购自西安尚诚生物科技有限公司。
青钱柳提取物为青钱柳(CYCLOCARYAPALIURUS)提取物,拉丁学名为CYCLOCARYAPALIURUS EXTRACT,购自上海珈得尔化学技术有限公司。
实施例和对比例的制备
缬草根提取物的制备(经过微生物发酵后的缬草根水提物)
步骤(1),取一定量缬草根切碎,加去离子水搅拌均匀,缬草根与去离子水的质量比为1:(1.0-1.3)(具体见表1所示),然后按每100g缬草根和水添加1-2g的添加量添加复配微生态制剂(枯草芽孢杆菌和凝结芽孢杆菌复配),常温浸渍发酵24-48h,无需密闭,过程中每间隔2h搅拌,得缬草根粗提物;
(2)将缬草根粗提物通过60-70目的滤网过滤过滤去渣后进行浓缩,浓缩至含水量为10-15%的膏状物,得缬草根提取物1-3。
表1缬草根提取物制备原料及参数
注:表中原料单位为质量份数,“——”表示未添加。
缬草根提取物的制备(未经过微生物发酵后的缬草根水提物)
步骤(1),取一定量缬草根切碎,加去离子水搅拌均匀,缬草根与去离子水的质量比为1:(1.0-1.3),然后在70℃下浸渍72h,过程中每间隔2h搅拌,得缬草根粗提物;
(2)将缬草根粗提物通过60-70目的滤网过滤过滤去渣后进行浓缩,浓缩至含水量为10-15%的膏状物,得缬草根提取物4、5,如表1所示。
紧致抗皱组合物的制备
按表2中质量份数取黑孢块菌提取物、芍药根提取物、缬草根提取物和青钱柳提取物进行混合,得紧致抗皱组合物1-7。
表2
注:表中“——”表示未添加
含有紧致抗皱组合物的精华液的制备
步骤(1),按表3质量份数取溶剂、螯合剂、增稠剂,进行混合搅拌,得混合物;
步骤(2),按表3质量份数取保湿剂、紧致抗皱组合物、棕榈酰三肽-5、羟丙基四氢吡喃三醇、pH调节剂、防腐剂按一定量加入混合物中,搅拌,得实施例和对比例。
表3实施例和对比例
表3实施例与对比例(续)
注:表中“——”表示未添加。
性能测试:
抑制弹性蛋白酶试验
本试验以猪胰弹性蛋白酶为测试对象,用N-琥珀酰-丙氨酸-丙氨酸-丙氨酸-p-硝基苯胺(AAAPVN)作为底物。猪胰弹性蛋白酶可以水解AAAPVN,其水解产物可引起420nm波长处吸光度增加,用酶标仪测定其吸光度,可评价实施例和对比例的紧致抗皱功效。
1.溶液配制
(1)待测样品:分别取实施例1-5和对比例1-5。
(2)配置Tris-HCl缓冲液(0.1M PH=8.0):称取2.42gTris于烧杯中,加入200mL的超纯水,溶解完全后用浓HCl调PH至8.0。
(3)阳性对照茶多酚溶液的配置(1mg/ml):称取5mg的茶多酚,用5ml Tris-HCl缓冲液进行溶解。
(4)配置底物溶液AAAPVN(2mM):称取4.51mg N-琥珀酰-丙氨酸-丙氨酸-丙氨酸-p-硝基苯胺,用5ml Tris-HCl缓冲液进行溶解。
(5)配置猪胰弹性蛋白酶液(0.171U/mL):取280μl溶于10ml Tris-HCl缓冲液中。
2.分组及加样
试验分为4组,分别是待测样品组、阳性对照组、空白对照组、模型对照组,同一组别同一浓度下设4个复孔。各溶液添加量如表4所示。
表4试验分组
3.测定方法
室温放置反应15min,用酶标仪在420nm处测定其吸光度。
4.计算式
式中:A0-空白对照孔吸光度的平均值;
A1-样品孔吸光度的平均值;
A2-模型对照孔吸光度的平均值。
5.实验结果
表5猪胰弹性蛋白酶抑制率结果
样品名称 | 弹性蛋白酶抑制率 |
模型对照 | 0.00% |
空白对照 | / |
阳性对照茶多酚(1mg/mL) | 70.52% |
实施例1 | 68.74% |
实施例2 | 71.14% |
实施例3 | 68.99% |
实施例4 | 68.20% |
实施例5 | 70.43% |
对比例1 | 53.05% |
对比例2 | 60.79% |
对比例3 | 56.33% |
对比例4 | 56.41% |
对比例5 | 64.30% |
促进I型胶原蛋白合成试验
本试验以成纤维细胞为检测对象,通过UVA辐照建立细胞光老化损伤模型,并检测待测样品作用后I型胶原蛋白(Collagen I)含量的变化,评价待测样品的紧致抗皱功效。
(1)接种:按1×104个/孔的接种密度接种细胞至96孔板,放置培养箱孵育过夜(37℃、5%CO2)。
(2)UVA照射:PBS清洗细胞后,根据实验分组,对有UVA照射的组别进行30J/cm2的UVA辐照。
(2)配液:分别选择实施例1-5和对比例1-5为待测样品溶液,以TGF-β(100ng/mL)为阳性对照进行实验。每孔加样100uL,每组设置3个复孔。给药完成后将96孔板放置在37℃培养箱中避光孵育24h。
(4)细胞上清收集:给药24h后取出96孔板,分别收集各样品组对应的细胞上清液至离心管中,1000rpm离心10min,收集上清置于1.5mL离心管中,-20℃储存备用。
(5)ELISA试剂盒测定细胞上清中I型胶原蛋白浓度:测试前30min取出试剂盒及待测样品,放置于室温后使用,测试操作严格按照试剂盒说明书操作。
表6 I型胶原蛋白含量检测结果
样品名称 | I型胶原蛋白含量均值(ng/mL) | I型胶原蛋白增长率 |
模型对照 | 1.304±0.129 | / |
空白(未受到UVA损伤) | 1.793±0.104 | 37.51% |
阳性(TGFβ1100ng/mL) | 1.579±0.090 | 21.11% |
实施例1 | 2.032±0.366 | 55.79% |
实施例2 | 2.114±0.211 | 62.03% |
实施例3 | 2.009±0.132 | 54.01% |
实施例4 | 2.020±0.306 | 54.13% |
实施例5 | 2.121±0.175 | 62.43% |
对比例1 | 1.804±0.101 | 38.02% |
对比例2 | 1.670±0.028 | 28.07% |
对比例3 | 1.921±0.031 | 47.10% |
对比例4 | 1.941±0.257 | 48.94% |
对比例5 | 1.549±0.020 | 18.80% |
抗氧化测试(DPPH自由基清除)
DPPH甲醇溶液为紫罗兰色,在517nm处有强的吸光值,若与待测样品结合,会降低在517nm处的吸光度值,由此判断样品清除DPPH自由基的能力。
(1)试验设计
DPPH用无水乙醇配制成2×10-4mol/L的溶液,实施例1-5和对比例1-5作为待检测液,制备待检测液、无水乙醇、DPPH溶液各2mL若干份,分别将混匀后室温下放置30分钟,于波长517nm处测定吸光度得Ai,每个样品平行测定3次,取平均值。
以Vc作为阳性对照,以无水乙醇代替DPPH测定吸光度值作为对照Aj空白,以DPPH溶液和无水乙醇混合测吸光度Ac。
按下式(1)计算各待测样品对DPPH自由基的清除率,并记录至下表7。
清除率(%)=(1-(Ai-Aj)/Ac)×100%...........(1)
式中:Ai为2mL实施例或对比例液+2mLDPPH试剂混合液的吸光度;Aj为2mL样品溶液+2mL无水乙醇混合液的吸光度;Ac为2mLDPPH溶液+2mL无水乙醇混合液的吸光度。
表7 DPPH清除率
组别 | DPPH清除率 |
实施例1 | 66.21% |
实施例2 | 70.11% |
实施例3 | 67.22% |
实施例4 | 65.42% |
实施例5 | 70.64% |
对比例1 | 53.73% |
对比例2 | 51.48% |
对比例3 | 60.18% |
对比例4 | 59.62% |
对比例5 | 43.17% |
结果分析
根据实施例1与对比例1、2性能测试结果相比可知,紧致抗皱组合物中的黑孢块菌提取物、芍药根提取物、缬草根提取物和青钱柳提取物之间具有共同促进皮肤修复的作用,协同作用下能够有效提高抑制弹性蛋白酶的活性的能力,提高促进皮肤胶原蛋白的合成的作用,增强对自由基离子的清除作用;通过本发明中优选的添加比例具有提高作用效果的好处。
根据实施例1与对比例3、4性能测试结果相比可知,缬草根通过复配微生态制剂发酵后能够有效的释放缬草根中的活性营养物质,复配微生态制剂在发酵中通过释放纤维素酶,来分解植物细胞壁,使得缬草根中的活性营养成分能够充分的溶解到溶剂中,提高了缬草根提取物的提取率。
最后所应当说明的是,以上实施例和对比例仅用以说明本发明的技术方案而非对本发明保护范围的限制,尽管参照较佳实施例对本发明作了详细说明,本领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或者等同替换,而不脱离本发明技术方案的实质和范围。
Claims (10)
1.一种含有紧致抗皱组合物的精华液,其特征在于,包括溶剂以及以下质量份数的组分:
紧致抗皱组合物1.5-2份;
棕榈酰三肽-5 0.3-1份;
羟丙基四氢吡喃三醇1.4-2.2份;
所述紧致抗皱组合物包括黑孢块菌提取物、芍药根提取物、缬草根提取物、青钱柳提取物。
2.根据权利要求1所述的含有紧致抗皱组合物的精华液,其特征在于,所述黑孢块菌提取物、芍药根提取物、缬草根提取物和青钱柳提取物的质量比为1:(0.23-0.57):(5.45-6.70):(2.64-3.11)。
3.根据权利要求1所述的含有紧致抗皱组合物的精华液,其特征在于,所述缬草根提取物制备方法包括以下步骤:
(1)取一定量缬草根切碎,加水搅拌均匀,然后添加复配微生态制剂,发酵24-48h后,得缬草根粗提物;
(2)将缬草根粗提物过滤去渣后浓缩,得缬草根提取物。
4.根据权利要求3所述的含有紧致抗皱组合物的精华液,其特征在于,所述步骤(1)中的缬草根与水的质量比为1:(1.0-1.3);所述复配微生态制剂的添加量为所述缬草根与水总质量的1-2%,复配微生态制剂中的微生物包括枯草芽孢杆菌和凝结芽孢杆菌,所述枯草芽孢杆菌的活菌数≥100亿CFU/g,凝结芽孢杆菌的活菌数≥50亿CFU/g;
所述步骤(2)中,缬草根粗提物通过≥60目的滤网过滤,然后浓缩至含水为10-15%的膏状物,得缬草根提取物。
5.根据权利要求1-4任一所述的含有紧致抗皱组合物的精华液,其特征在于,还包括但不限于以下组分:螯合剂、增稠剂、保湿剂、pH调节剂、防腐剂。
6.根据权利要求5所述的含有紧致抗皱组合物的精华液,其特征在于,所述增稠剂为羟丙基甲基纤维素、黄原胶、叔丁醇中的至少一种。
7.根据权利要求5所述的含有紧致抗皱组合物的精华液,其特征在于,所述保湿剂为透明质酸钠、甜菜碱、甘油、丁二醇、赤藓醇中的至少一种。
8.根据权利要求5所述的含有紧致抗皱组合物的精华液,其特征在于,所述防腐剂为丙二醇、辛酰羟肟酸、1,2-己二醇、乙基己基甘油、对羟基苯乙酮中的至少一种。
9.根据权利要求5所述的含有紧致抗皱组合物的精华液,其特征在于,所述螯合剂为EDTA二钠、EDTA四钠中的至少一种;
所述pH调节剂为氨甲基丙醇、柠檬酸钠、氢氧化钠中的至少一种。
10.根据权利要求5所述的含有紧致抗皱组合物的精华液的制备方法,其特征在于,包括以下步骤:
步骤(1),将所述溶剂、螯合剂、增稠剂按一定量进行混合搅拌,得混合物;
步骤(2),将所述保湿剂、紧致抗皱组合物、棕榈酰三肽-5、羟丙基四氢吡喃三醇、pH调节剂、防腐剂按一定量加入混合物中,搅拌,得所述含有紧致抗皱组合物的精华液。
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