CN117298021B - 一种具有抗炎舒缓作用的组合物及其制备方法和应用 - Google Patents
一种具有抗炎舒缓作用的组合物及其制备方法和应用 Download PDFInfo
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- CN117298021B CN117298021B CN202311595897.2A CN202311595897A CN117298021B CN 117298021 B CN117298021 B CN 117298021B CN 202311595897 A CN202311595897 A CN 202311595897A CN 117298021 B CN117298021 B CN 117298021B
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- buddleja officinalis
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- bisabolol
- panthenol
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- Cosmetics (AREA)
Abstract
本申请提供了一种具有抗炎舒缓作用的组合物及其制备方法和应用，本申请的组合物包含密蒙花提取物、桃金娘果提取物、雪莲花提取物、山布枯提取物、紫苏籽油以及泛醇、红没药醇、硫辛酸。本申请还提供了该组合物的制备方法以及该组合物在具有抗炎、舒缓和保湿等作用的轻医美产品和护肤品中的应用。
Description
技术领域
本申请属于天然植物提取物领域和化妆品领域，具体地，本申请提供了一种具有抗炎舒缓作用的组合物及其应用。
背景技术
在化妆品领域中，天然、绿色、健康无论在研发还是宣传中都是热点概念，因此各种植物提取物也属于化妆品中的常用成分，多年的研究和应用使得本领域对常见植物提取物的效果和制备方法等有了比较清楚的认识。
目前包含植物提取物的化妆品用组合物的发展方向主要在于新植物提取物的开发和功能鉴定，以及现有植物提取物提取方法和改进有有效成分的鉴定。
发明内容
针对上述情况，申请人使用有消炎、舒缓、抑菌、控油、保湿效果的密蒙花提取物、桃金娘果提取物、雪莲花提取物、山布枯提取物、紫苏籽油植物提取物制备了可用于多种化妆品的组合物，其中加入的泛醇、红没药醇、硫辛酸进一步强化了其效果，申请人对植物提取物中目前研究较少的密蒙花提取物制备过程进行了进一步研究，优化提高了其中主要有效成分的提取得率并改善了其舒缓和抑菌效果。
一方面，本申请提供了一种具有抗炎舒缓作用的组合物，所述组合物包含密蒙花提取物、桃金娘果提取物、雪莲花提取物、山布枯提取物、紫苏籽油以及泛醇、红没药醇、硫辛酸。
进一步地，所述组合物中密蒙花提取物、桃金娘果提取物、雪莲花提取物、山布枯提取物、紫苏籽油、泛醇、红没药醇、硫辛酸重量比例为30-60：10-30：10-30：5-20：5-20：5-10：1-5：1-3。
进一步地，所述组合物中密蒙花提取物、桃金娘果提取物、雪莲花提取物、山布枯提取物、紫苏籽油、泛醇、红没药醇、硫辛酸重量比例为40：20：20：10：10：10：2：1。
进一步地，所述密蒙花提取物通过以下方法制备：
（1）粉碎得密蒙花粉末；
（2）使用纤维素酶和碱性蛋白酶酶解；
（3）乙醇回流提取、旋转蒸发并冻干。
进一步地，步骤（2）为：将（1）得到的密蒙花粉末加入2-8倍质量的，含300-800U/mL纤维素酶和100-500U/mL碱性蛋白酶和螯合剂的水溶液中，调节pH至7-9；40℃下酶处理0.5-2.5小时；过滤，得到滤渣。
进一步地，所述螯合剂为EDTA-Na2或者EDDS-Na3。
进一步地，步骤（2）为：将（1）得到的密蒙花粉末加入5倍质量的，含500U/mL纤维素酶和200U/mL碱性蛋白酶和0.2 mg/mL EDDS-Na3的水溶液中，调节pH至7-8；40℃下酶处理1.5-2.5小时；过滤，得到滤渣。
进一步地，步骤（3）为：以（1）得到的密蒙花粉末10倍质量的70%的乙醇80℃回流提取步骤（2）得到的滤渣2次；合并提取液，旋转蒸发浓缩后，冷冻干燥制成密蒙花提取物。
另一方面，本申请提供了上述组合物在制备具有抗炎、控油、舒缓和保湿作用的化妆品中的应用。
进一步地，所述化妆品为乳液、霜膏或者面膜。
进一步地，所述化妆品中还包含化妆品中可接受的辅料。
除上述剂型外，本申请的组合物也可以用于制备其他化妆品剂型，如化妆水、护肤油等，本领域技术人员可以根据溶解、分散、乳化、稳定等需要为其选择适合的辅料。
本申请中所述的化妆品中可接受的辅料，根据国家、剂型等因素的不同而有所区别，本领域技术人员可以通过查阅相关规定和资料来准确选择。
附图说明
图1为斑马鱼胚胎舒缓试验的结果，图中1-6依次为无酶解、500U/mL纤维素酶+200U/mL碱性蛋白酶、500U/mL纤维素酶+200U/mL碱性蛋白酶+0.5mg/mL醋酸钠、500U/mL纤维素酶+200U/mL碱性蛋白酶+0.2mg/mL EDTA-Na2、500U/mL纤维素酶+200U/mL碱性蛋白酶+0.5 mg/mL EDDHA-Na、500U/mL纤维素酶+200U/mL碱性蛋白酶+0.2 mg/mL EDDS-Na3组的结果。
具体实施方式
实施例1 主要材料和实验方法
主要材料：
密蒙花药材购自正规中药材供应商，产地标称湖北，经中药药材专业人员鉴定为密蒙花干燥花序部分，实验中使用同一批次购买的药材；
纤维素酶、果胶酶：食品级，夏盛；
碱性蛋白酶、木瓜蛋白酶：食品级，华上翔洋；
EDDS-Na3（乙二胺二琥珀酸三钠）、EDDHA-Na（乙二胺二邻苯基乙酸钠）：长信化工；
EDTA-Na2（乙二胺四乙酸二钠）、醋酸钠：国药；
毛蕊花苷和蒙花苷的检测：
仪器：安捷伦1220 Infinity II；配Eclipse XDB-C18柱；
色谱条件：流动相A为乙腈，流动相B为纯净水，梯度洗脱A 20% 10min、A 40%10min、A 60% 20min；检测波长327nm，流速1.5mL/min，柱温27℃；
毛蕊花苷和蒙花苷标准品：成都钠钶锂生物技术有限公司，纯度大于99%；
使用标注品配合浓度梯度溶液，检测考察线性关系后，外标法检测3份平行样品的毛蕊花苷和蒙花苷含量。
密蒙花提取物的制备：
参考现有技术，提取使用回流提取方法，以10倍质量的70%的乙醇80℃回流提取2次，合并提取液旋转蒸发除去乙醇并浓缩后，冷冻干燥制成提取物粉末（按照现有技术记载，醇提法对黄酮类和苯乙醇苷等极性成分提取率较好，提取率达到80%以上）。
DPPH自由基清除能力检测：
将待测提取物配制成10mg/mL水溶液；取水溶液样品2.5mL，与2.5mL 0.1mM的DPPH无水乙醇溶液（测试）或者2.5mL无水乙醇混合（对照），25℃下避光反应30min；以分光光度计测定样品反应液、对照反应液以及参比（2.5mL无水乙醇加2.5mL 0.1mM的DPPH无水乙醇溶液）517nm处的吸光度，通过以下公式计算，DPPH自由基清除率%=（1-（A测试-A对照）/A参比）（三份平行试验取平均值）。
斑马鱼舒缓行为实验：
参照广州开发区化妆品产业协会制定的一系列斑马鱼胚胎测试方法征求意见稿进行：使用 96 孔细胞培养板为实验载体，每孔加入胚胎 5 枚（授精后36h左右的野生型AB 系斑马鱼胚胎，3份重复）；吸去养殖用水并清洗后，向空白组和模型组中每孔加入100uL水，样品组每孔分别中加入 100uL 一定浓度（经过预实验确定为1%w/v）的受试物溶液，盖上培养板面板，在 28.0±0.5℃生化培养箱中孵育 1h；1h 后，每孔分别加入 100uL造模剂(500uM十二烷基硫酸钠溶液)，在28.0±0.5摄氏度生化培养箱中孵育 15min 后，录制 30s 视频,统计每组胚胎自旋运动次数，相对刺激强度=样品组或模型组胚胎自旋运动次数/空白组胚胎自旋运动次数。
抑菌浓度实验
将提取物配制成0.1%、0.2%、0.5%、1%、2%、5%、10%、20% w/v的水溶液；以水为阴性对照。取1×107 CFU/mL的菌悬液1mL，加入水溶液1mL以及对应液体培养基1.5mL；混合均匀后37摄氏度培养48小时；观察抑菌情况。使用白色念珠菌ATCC 10231、绿脓杆菌ATCC15442、金黄色葡萄球菌ATCC 29213的标准菌株传代所得菌株作为测试菌。
酶处理方法
向粉碎后的密蒙花粉末中加入5倍质量的，含一定量的酶和其他成分的水溶液，调节pH至7.5；40℃下酶处理2小时；过滤，滤渣加水高温灭酶后储存或者直接用于提取物制备（提取过程的乙醇和高温已经可以实现灭活酶的效果）。
实施例2 密蒙花提取物的制备
按照实施例1所述的方法，制备密蒙花提取物，其中毛蕊花苷和蒙花苷得率（按照提取物中的含量/原料重量计算）为2.45±0.11%和1.03±0.04%，提取物具备一定的抗氧化、舒缓和抗菌效果（具体数据参见以下的实施例）。
为进一步提高提取效果，尝试了在提取前加入酶进行处理，组合实验和用量梯度实验结果表明（具体数据未展示），纤维素酶和碱性蛋白酶的组合（向粉碎后的密蒙花粉末中加入5倍质量的，含500U/mL纤维素酶和200U/mL碱性蛋白酶水溶液，调节pH至7.5，酶处理2小时；过滤，滤渣直接用于提取物制备）能最有效地提高了毛蕊花苷和蒙花苷得率，分别达到了3.43±0.08%和1.31±0.03%，随着黄酮类和苯乙醇苷等极性成分得率提高，提取物产品的抗氧化、舒缓效果也得到了增强（具体数据参见以下的实施例）。
实施例3酶解体系的进一步优化
由于密蒙花的药用部位不同于大多数中药材的根、茎、叶、果实等，除常规酶解方式外，我们还尝试了加入表面活性剂、螯合剂等成分以改变酶解产物的分散方式或者去除与膜蛋白结合的金属以暴露酶解敏感部分。结果表明加入螯合剂可以有效提高主要有效成分得率并增强提取物的抗氧化、舒缓效果（优选浓度的梯度实验结果未展示）。得率和DPPH自由基清除率如表1所示，舒缓效果的变化如图1所示，抑菌浓度的变化如表2所述。
表1不同酶解方式对有效成分得率和提取物DPPH自由基清除率的影响
注：用量不计水合物中的水。
表2不同酶解方式对抑菌效果的影响
注：用量不计水合物中的水。
结果表明，加入螯合剂确实提高了毛蕊花苷和蒙花苷的得率，某些方法提高蒙花苷得率接近30%（对毛蕊花苷得率的效果不及蒙花苷显著）。但DPPH自由基清除率、抑菌效果和舒缓效果的改善与毛蕊花苷和蒙花苷得率的改善不完全一致，相比其他组，加入EDDS-Na3明显提高了抑菌效果和舒缓效果，而加入EDTA-Na2对抗氧化效果改善比较有利。这表明这些效果可能与提取物中的多种黄酮、萜类、挥发油类甚至蛋白和糖成分有关。
实施例4 乳液组合物的制备
使用实施例3获得的密蒙花提取物（500U/mL纤维素酶+200U/mL碱性蛋白酶+0.2mg/mL EDDS-Na3组），配合桃金娘果提取物、雪莲花提取物、山布枯提取物、紫苏籽油以及泛醇、红没药醇、硫辛酸制备了乳液组合物，其中含密蒙花提取物2%、桃金娘果提取物1%、雪莲花提取物1%、山布枯提取物0.5%、紫苏籽油0.5%、泛醇0.5%、红没药醇0.1%、硫辛酸0.05%（固体、液体均按照w/v计算）以及甘油、角鲨烷、丁二醇、甘油聚醚26等辅料。
初步的家兔皮肤动物实验和志愿者试用表明，上述乳液产品无明显不良气味和刺激性，并且具有抗炎、控油、舒缓和保湿效果。
Claims (4)
1.一种组合物在制备具有抗炎、控油、舒缓和保湿作用的化妆品中的应用，其特征在于，所述组合物包含密蒙花提取物、桃金娘果提取物、雪莲花提取物、山布枯提取物、紫苏籽油以及泛醇、红没药醇、硫辛酸；所述组合物中密蒙花提取物、桃金娘果提取物、雪莲花提取物、山布枯提取物、紫苏籽油、泛醇、红没药醇、硫辛酸重量比例为30-60：10-30：10-30：5-20：5-20：5-10：1-5：1-3；
所述密蒙花提取物通过以下方法制备：
（1）粉碎得密蒙花粉末；
（2）将（1）得到的密蒙花粉末加入5倍质量的，含500U/mL纤维素酶和200U/mL碱性蛋白酶和0.2 mg/mL EDDS-Na3的水溶液中，调节pH至7-8；40℃下酶处理1.5-2.5小时；过滤，得到的滤渣用于步骤（3）的提取；
（3）乙醇回流提取、旋转蒸发并冻干。
2.根据权利要求1所述的应用，其中所述组合物中密蒙花提取物、桃金娘果提取物、雪莲花提取物、山布枯提取物、紫苏籽油、泛醇、红没药醇、硫辛酸重量比例为40：20：20：10：10：10：2：1。
3.根据权利要求1或2所述的应用，其中，步骤（3）为：以（1）得到的密蒙花粉末10倍质量的70%的乙醇80℃回流提取步骤（2）得到的滤渣2次；合并提取液，旋转蒸发浓缩后，冷冻干燥制成密蒙花提取物。
4.根据权利要求1所述的应用，其中所述化妆品为乳液、霜膏或者面膜。
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