CN114425030A - 一种植物提取物组合物、制备方法及其用途 - Google Patents
一种植物提取物组合物、制备方法及其用途 Download PDFInfo
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- CN114425030A CN114425030A CN202210139706.0A CN202210139706A CN114425030A CN 114425030 A CN114425030 A CN 114425030A CN 202210139706 A CN202210139706 A CN 202210139706A CN 114425030 A CN114425030 A CN 114425030A
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- Prior art keywords
- moringa seed
- extract
- moringa
- glabridin
- tyrosinase
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- Medicines Containing Plant Substances (AREA)
Abstract
本发明公开了一种植物提取物组合物及其用途,包括辣木籽提取物和光甘草定,且辣木籽提取物为0.25‑3mg/mL,光甘草定为0.25‑5mg/mL;所述的辣木籽提取物的制备方法为:(1)将辣木籽原料,加去离子水,超声萃取,取上清液,过滤、静置、离心、旋转蒸干,得辣木籽水提物;(2)将辣木籽原料,加30%‑70%的乙醇,超声萃取,取上清液,过滤、静置、离心、旋转蒸干,得辣木籽油提物;(3)将辣子水提物和辣木籽油提物混合,得辣木籽提取物。该复配剂在0.25mg/mL时其对酪氨酸酶的抑制效果就远远超过光甘草定和辣木提取物单独使用的效果,而且优于常用的熊果苷和烟酰胺的效果。
Description
技术领域
本发明涉及化妆品技术领域,尤其涉及一种植物提取物组合物及其用途。
背景技术
通过抑制黑色素的合成来达到美白效果是当今美白产品发展的方向。目前, 熊果苷、曲酸、维生素C以及它们的衍生物等已被广泛应用于美白产品中,取 得了一定的效果。但是,上述美白活性成分的渗透率较低,功效不够理想,配 方师往往通过添加大量美白剂来增强效果,但仍难以满足消费者的需求。同时, 随着添加量增大,产生了许多安全隐患。例如,研究表明长期应用曲酸会引起 接触性皮炎和红斑反应,并可能诱发妇女乳腺癌;熊果苷则是通过释放氢醌来 达到美白效果,而后者的细胞毒性很大,已被禁止应用于化妆品中。这些成分 还容易导致化妆品的质量问题:熊果苷应用于配方中易引起产品变色;曲酸会 与许多金属离子螯合,特别是的铁复离合子物产,与生空着气色接触缓慢氧化, 在高温下还能加速。维生素C及其衍生物存在容易氧化或吸收困难等缺点。为此, 研究和开发安全性好、质量稳定、能满足消费者功效要求的美白产品,已成为 化妆品业界亟需解决的热门课题。
光甘草定(4-[(3R)-8,8-Dimethyl-3,4-dihydro-2H-pyrano[6,5-f] chromen-3-yl]Ben-Zene-1,3-diol)从特定品种的甘草中提取,是目前世界上 公认最有效的天然美白剂。它能抑制酪氨酸酶的活性,又能抑制多巴色素互变 和DHICA氧化酶的活性,是一种快速、高效、绿色的美白祛斑化妆品添加剂,具 有与SOD(过氧化物歧化酶)相似的清除氧自由基的能力,以及与维生素E相近 的抗氧自由基能力,还具有抗菌消炎、抗氧化、抗衰老、吸收紫外线、美白亮 肤、祛斑的作用。光甘草定目前已在化妆品行业大量使用,如在膏霜、水、露、 乳液等产品中都得到广泛应用。
辣木为多年生热带落叶乔木,原产于印度,因其根具有辛辣味而得名。辣 木又称为“鼓槌树”“奇迹之树”,是粮食安全新资源作物,对人类健康有重 要作用。辣木叶其主要组成为蛋白质(约含37%)、油脂(约含34%),还含有丰 富的矿质元素、纤维素、多糖、维生素、甾醇、氨基酸、多酚等,研究表明, 辣木叶具有抗衰、美白、活血、调节免疫系统、抗紫外线、抗肿瘤、净化水质 等作用。从已报道的文献来看,黄酮、生物碱、硫代葡萄糖苷、异硫氰酸酯、 酚及酚酸等化合物是辣木发挥药理作用的主要活性成分。高秋玉等研究发现, 辣木叶乙醇提取物中,以新绿原酸为代表的总奎宁酸和以异槲皮苷代表的总黄 酮类成分对1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl radical,DPPH)清除率高达80%,具有明显的抗氧化作用。TUORKEY等研究发 现,辣木叶水提物可改善四氧嘧啶诱导的糖尿病小鼠胰岛素抵抗,增加总抗氧 化能力,提高免疫耐受性。
辣木提取物与光甘草定在美白方面各自有其独到的优势,两者的抑制机理 各有不同,但价格都相当昂贵,应用起来成本压力很大,因而难以推广。如果 将其加以复配,就能够发挥协同效应,从而既能够增强美白效果,还可以降低 生产成本,生产出满足社会需要的美白产品,并产生巨大的经济和社会效益。
发明内容
本发明一个目的在于公开了一种植物提取物组合物,其包括辣木籽提取物 和光甘草定。
本发明的另一目的在于公开了该植物提取物组合物制备酪氨酸酶抑制剂的 用途。具体技术方案如下:
一种植物提取物组合物,包括辣木籽提取物和光甘草定,且辣木籽提取物 为0.25-3mg/mL,光甘草定为0.25-5mg/mL;
所述的辣木籽提取物的制备方法为:(1)将辣木籽原料,加去离子水,超 声萃取,取上清液,过滤、静置、离心、旋转蒸干,得辣木籽水提物;(2)将 辣木籽原料,加30%-70%的乙醇,超声萃取,取上清液,过滤、静置、离心、旋 转蒸干,得辣木籽油提物;(3)将辣子水提物和辣木籽油提物混合,得辣木籽 提取物。
进一步的,所述的将辣子水提物和辣木籽油提物混合的质量比为1:1。
更进一步的,所述的辣木籽提取物为0.25-2mg/mL,光甘草定为0.25-1mg/mL。
更进一步的,所述的辣木籽原料经烘干、粉碎、过40-80目筛后,成为辣 木籽粉末。
更进一步的,所述的辣木籽粉末中加入去离子水,辣木籽粉末与去离子水 料液比为1:12,超声提取温度20-40℃,提取30-60min,超声功率为1000W, 工作间歇比2s/2s、循环泵转速25r/s。
更进一步的,辣木籽粉末中加入70%乙醇,辣木籽粉末与70%乙醇料液比为 1:8,超声提取温度为室温,提取1h,超声功率为1000W,工作间歇比2s/2s、 循环泵转速25r/s。
一种上述植物提取物组合物制备酪氨酸酶抑制剂的用途。
更进一步的,所述的植物提取物组合物在制备护肤品或化妆品中的用途。
本发明的有益效果在于,与现有技术相比,目前的辣木提取物和光甘草定 单独使用时其对酪氨酸酶有一定的抑制作用,但使用量较大,一方面价格昂贵 影响其广泛使用,另一方面当植物提取物含量较大时,影响化妆品的稳定性, 本发明中将光甘草定和辣木提取物进行复配,在0.25mg/mL时其对酪氨酸酶的 抑制效果就远远超过光甘草定和辣木提取物单独使用的效果,而且优于常用的 熊果苷和烟酰胺的效果。
附图说明
图1为本发明辣木籽提取物和光甘草定对酪氨酸酶活力的影响。
图2为不同辣木籽提取物质量浓度与酪氨酸酶抑制率的关系。
图3为辣木籽提取物对酪氨酸酶催化L-酪氨酸抑制作用的Linweaver-Bulk 曲线。
图4为不同质量浓度的光甘草定对酪氨酸酶的抑制作用关系图。
图5为光甘草定对酪氨酸酶催化L-酪氨酸抑制作用的Linweaver-Bulk曲 线。
具体实施方式
下面将结合本发明实施例中的附图,对本发明实施例中的技术方案进行清 楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是 全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有作出创造 性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
实施例1:辣木籽提取物的制备
(1)辣木籽水提取物的制备:
将辣木籽80℃烘干,粉碎,过50目筛,按照辣木籽粉末与去离子水1:12 的液料比加入去离子水,30℃下进行超声提取40min,超声功率为1000W,工作 间歇比2s/2s、循环泵转速25r/s,静置后取上清液,纱布过滤,将滤液静 置,再将上层清液离心,离心转速为3000-4000r/min,离心时间为10min,取 上清液进行真空旋转蒸发,真空旋转蒸发的真空度为0.07-0.09MPa,即得辣木 籽水提物。
(2)辣木籽油提物的制备:
将辣木叶80℃烘干,粉碎,过50目筛,按照辣木籽粉末与70vol%乙醇1:8 的液料比加入70vol%乙醇,室温下进行超声提取1h,超声功率为1000W,工作 间歇比2s/2s、循环泵转速25r/s,静置后取上清液,纱布过滤,将滤液静 置,再将上层清液离心,离心转速为3000-4000r/min,离心时间为10min,取 上清液进行真空旋转蒸发,真空旋转蒸发的真空度为0.07-0.09MPa,即得辣木 籽油提物。
(3)将辣子水提物和辣木籽油提物混合,充分混合均匀,混合质量比为1:1, 得辣木籽提取物。
实施例2:测试样品对酪氨酸酶活性抑制率
采用多巴速率法测定样品对酪氨酸酶活性的抑制作用。
按照表1中所示,将L-酪氨酸、样品液、PBS缓冲液依次加到编号为A1、A2、B1、B2组各管中、混匀:将试管架放入37℃恒温水浴锅中水浴10-15min: 将酷氨酸酶依次加入A1和B1组各管中、摇匀,准确计时,反应15min停止: 依次在96孔板上点板,用酶标仪测定其吸光度并计算抑制率。抑制率计算 公式为:
式中,A1为空白样板有酶体系吸光度;A2为空白样板无酶体系吸光度;B1为样品组有酶体系吸光度;B2为样品组无酶体系吸光度。
实验中阴性对照用PBS缓冲液代替,阳性对照选用质量分数为0.5%的α- 熊果苷水溶液,每组平行测定6次。
表l反应液组成
不同质量浓度辣木籽提取物和光甘草定对酪氨酸酶的抑制作用。采用多巴 速率法测定样品对酪氨酸酶活性的抑制作用,向反应体系中分别加入不同质量 浓度的辣木籽提取物和光甘草定,依次为0.1、0.25、0.5、1、1.5、2、3、4、 5、6mg/mL,测定反应体系在475nm处的吸光度,计算辣木籽提取物和光甘草 定对酪氨酸酶的抑制率,并根据浓度-酶抑制曲线估计辣木籽提取物和光甘草定 的半抑制浓度IC50,见图1,辣木籽提取物和光甘草定对酪氨酸酶活力的影响。 光甘草定购自陕西绿莱生物有限公司。
由图1可知,随着光甘草定质量浓度的增加,对酪氨酸酶的抑制率也增加, 当其质量浓度达到一定时,对酪氨酸酶的抑制率保持不变,且维系在一个较大 值。辣木籽提取物的浓度在0.1-2.0mg/mL时,对酪氨酸酶的抑制率会随着提取 液质量浓度的升高而增大,表现为剂量依赖性抑制。但随着质量浓度继续增大, 抑制率反而下降,说明辣木籽提取物对酪氨酸的抑制率与其浓度并不呈现单一 的增减关系。其中,光甘草定的IC50为4.43mg/mL,辣木籽提取物的IC50为 1.4mg/mL。
实施例3:辣木籽提取物对酪氨酸酶抑制作用的动力学分析。
抑制剂与酶的作用方式分为可逆抑制作用与非可逆抑制作用,其中可逆抑 制作用又分为竞争型、非竞争型、反竞争性型和线性混合型,可通过 Lineweaver-Burk双倒数图来判断。
固定酪氨酸酶的浓度不变,采用多巴速率法测定样品对酪氨酸酶活性的抑 制作用,见实施例2,分别测L-酪氨酸浓度为0.5、1、1.5、2mmol/L时,质 量浓度为0.1、0.25、0.5、1mg/mL的辣木籽提取物的吸光度,绘制浓度-ΔA曲 线,见图2,图2辣木籽提取物质量浓度与酪氨酸酶抑制率的关系。
通过吸光度的改变值求初速度V0(1min内吸光度的变化值)。以底物L- 酪氨酸浓度的倒数值为横坐标,以初速度V0的倒数值为纵坐标作图,得到图3, 辣木籽提取物对酪氨酸酶催化L-酪氨酸抑制作用的Linweaver-Bulk曲线。由图 3可知,随着底物L-酪氨酸浓度增大,V0随之增大。不同质量的辣木籽提取物的 直线均与无提取液的直线相交于第二象限,且直线的横截距减小,纵截距增大, 即Michealis-Menten动力学方程的Vmax减小,Km增大,表明辣木籽提取物对 酪氨酸酶的抑制类型为线性混合型抑制,即辣木籽提取物和L-酪氨酸与酪氨酸 酶的结合互不相关,既不排斥也不促进,但酶与底物L-酪氨酸或辣木籽提取物 的反应常数Km会因结合次序的不同而改变。
实施例4:光甘草定对酪氨酸酶抑制作用的动力学分析。
由实施例2可知,当光甘草定浓度大于2mg/mL时,抑制率升高趋势平缓, 故选2mg/mL的4个浓度(2.0mg/mL、3.0mg/mL、4.0mg/mL、5.0mg/mL) 进行动力学实验。固定底物L-酪氨酸酶浓度为1.0mmol/L,改变酶含量(40、 60、80和100U/mL)和光甘草定浓度(2.0mg/mL、3.0mg/mL、4.0mg/mL、5.0 mg/mL),测定吸光值A475,结果见图4,不同质量浓度的光甘草定对酪氨酸酶的 抑制作用关系图。图4显示,在相同酶活力下,随着光甘草定浓度的增高,A475吸光值逐渐变小,直线的斜率下降,说明光甘草定对酪氨酸酶的作用属于可逆 抑制。
由上述结果可知,光甘草定对酪氨酸酶活性的抑制属于可逆抑制。可逆抑 制分为竞争型、非竞争型、反竞争性型和线性混合型。固定底物酪氨酸酶含量 (100U/mL),改变L-酪氨酸酶浓度(0.25、0.5、0.75和1.0mmol/L)和光甘草 定浓度(2.0mg/mL、3.0mg/mL、4.0mg/mL、5.0mg/mL),测定吸光值A475, 通过Lineweaver-Burk取双倒数图判断抑制类型,结果如图5所示,光甘草定抑 制酪氨酸酶动力学曲线图。图5显示,Lineweaver-Burk双倒数作图相交于横轴 的一点,A475吸光值随着光甘草定的浓度增加而变小,抑制常数Km不变,反应速 度随光甘草定浓度增加而减小,复合非竞争性抑制类型的特征。光甘草定对酪 氨酸酶的抑制作用通过降低酶的活力,与酪氨酸酶分子上的独立部位结合,而 不与底物L-酪氨酸竞争活性中心。
实施例5:复配使用辣木籽提取物与美白剂对酪氨酸酶的抑制率
不同质量浓度的辣木籽提取物分别与3种美白剂(熊果苷、光甘草定和烟 酰胺)进行复配,测得酪氨酸酶活性抑制率见表2,不同浓度的辣木籽提取物与 三种美白剂复配时对酪氨酸酶的抑制率。表2显示,熊果苷与辣木籽提取物复 配时,7个浓度梯度间的酪氨酸酶抑制率均无统计学差异(P>0.05),表明辣木 提取物不能促进熊果苷对酪氨酸酶活性的抑制率。当辣木籽提取物与光甘草定 复配时,7个浓度梯度间的酪氨酸酶抑制率均明显上升(P<0.01),表明辣木籽 提取物与光甘草定复配对酪氨酸酶的抑制作用具有协同促进效果。辣木籽提取 物与烟酰胺复配时,7个浓度梯度间的酪氨酸酶抑制率均明显上升(P<0.01), 表明辣木提取物与烟酰胺复配时,对酪氨酸酶的抑制作用具有协同促进效果。 此外,辣木籽提取物对光甘草定的促进作用总体上优于烟酰胺的促进作用。这 可能与这两种成分与酪氨酸酶作用的动力学机理有关。
实施例6:辣木籽提取物与光甘草定复配比例。
从实施例5可以看出当固定辣木籽提取物的浓度为2mg/mL时,随着光甘草 定浓度的增加当光甘草定浓度为1.0mg/mL时对酪氨酸酶的抑制率已经达到了 100%,因此,取0-2mg/mL的辣木籽提取物,0-1mg/mL的光甘草定进行复配,测 定其对酪氨酸酶的协同抑制作用。
从表2和表3可知,当辣木籽提取物在0-2mg/mL时与0-1mg/mL的光甘草 定进行复配对酪氨酸酶具有很好的抑制作用。两者进行复配在0.25mg/mL时其 对酪氨酸酶的抑制作用远远超过两者单独使用的抑制作用。因此,将两者进行 复配将大大降低各自原料的使用量,并且能达到较强的酪氨酸酶抑制作用。
以上所述是本发明的优选实施方式,应当指出,对于本技术领域的普通技 术人员来说,在不脱离本发明原理的前提下,还可以做出若干改进和润饰,这 些改进和润饰也视为本发明的保护范围。
Claims (8)
1.一种植物提取物组合物,其特征在于,包括辣木籽提取物和光甘草定,且辣木籽提取物为0.25-3mg/mL,光甘草定为0.25-5mg/mL;
所述的辣木籽提取物的制备方法为:(1)将辣木籽原料,加去离子水,超声萃取,取上清液,过滤、静置、离心、旋转蒸干,得辣木籽水提物;(2)将辣木籽原料,加30%-70%的乙醇,超声萃取,取上清液,过滤、静置、离心、旋转蒸干,得辣木籽油提物;(3)将辣子水提物和辣木籽油提物混合,得辣木籽提取物。
2.如权利要求1所述的植物提取物组合物,其特征在于,所述的将辣子水提物和辣木籽油提物混合的质量比为1:1。
3.如权利要求2所述的植物提取物组合物,其特征在于,所述的辣木籽提取物为0.25-2mg/mL,光甘草定为0.25-1mg/mL。
4.如权利要求3所述的植物提取物组合物,其特征在于,所述的辣木籽原料经烘干、粉碎、过40-80目筛后,成为辣木籽粉末。
5.如权利要求3所述的植物提取物组合物,其特征在于,所述的辣木籽粉末中加入去离子水,辣木籽粉末与去离子水料液比为1:12,超声提取温度20-40℃,提取30-60min,超声功率为1000W,工作间歇比2s/2s、循环泵转速25r/s。
6.如权利要求3所述的植物提取物组合物,其特征在于,辣木籽粉末中加入70%乙醇,辣木籽粉末与70%乙醇料液比为1:8,超声提取温度为室温,提取1h,超声功率为1000W,工作间歇比2s/2s、循环泵转速25r/s。
7.一种如权利要求1-6中任一项植物提取物组合物制备酪氨酸酶抑制剂的用途。
8.如权利要求7所述的用途,其特征在于,所述的植物提取物组合物在制备护肤品或化妆品中的用途。
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