CN114848721B - 一种含植物提取物的祛痘组合物及其应用 - Google Patents
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Abstract
本发明公开了一种含植物提取物的祛痘组合物及其应用,旨在提供一种具有祛痘消肿,抑菌,舒缓舒敏,去痘印疤痕等作用的含植物提取物的祛痘组合物;其技术方法由以下重量份的组分制成:粉防己提取物60‑90份,皮肤软化剂1‑20份;黄柏提取物5‑30份;龙胆提取物5‑20份;属于日化技术领域。
Description
技术领域
本发明公开了一种祛痘组合物,具体地说,是一含植物提取物的祛痘组合物,本发明还涉及该组合物的制备方法与应用,属于日化领域。
背景技术
痤疮,俗名青春痘,是一种发生在毛囊皮脂腺及其周围组织的炎症病变。痤疮的发生主要与皮脂分泌过多、毛囊皮脂腺导管阻塞、细菌感染和炎症反应等诸多因素密切相关。寻常痤疮的发病机理为:面部皮脂分泌增多,导致毛囊和皮脂腺导管角化过度。角化过度的细胞和皮脂结合,淤积于毛囊形成脂肪粒,脂肪粒被毛囊中的痤疮丙酸杆菌分解,产生游离脂肪酸,引起炎症形成痤疮。
当前市场祛痘类护肤品主要分为2大类型,一类以抗生素,维A 酸类等西药为主,见效快,但是容易反复,容易发生耐药性,甚至因为西药的高浓度容易导致刺激性,这类药物的不良反应和副作用较为棘手,一类以中药添加为主,但是存在添加成分粗糙,见效慢,外观粗糙,色泽过深容易导致皮肤色素沉着问题。
目前市场上使用天然植物提取物的祛痘护肤品,其有效物含量及祛痘功效的作用情况没有科学严谨的数据证明。大多数天然植物提取物的添加仅仅停留在“概念性”添加的水平上,祛痘产品本身是否具有显著功效以及祛痘护肤品对人体是否安全无法得到保障。另外,目前市场上常见的含有抗菌杀菌成分、去角质成分以及控油成分的传统祛痘护肤品,虽然对青春痘有一定的治疗效果,但对使用部位有明显的刺激和损伤并且长期使用,容易产生抗药性,并且可能对人体免疫系统带来不可估量的损害。
防己,是防己科植物粉防己Stephania tetrandra S.Moore的干燥根。秋季采挖,洗净,除去粗皮,晒至半干,切段,个大者再纵切,干燥;苦,寒。归膀胱、肺经。祛风止痛,利水消肿。用于风湿痹痛,水肿脚气,小便不利,湿疹疮毒。
作为经典药材,粉防己在传统皮肤外科应用较少,但近年来广州伽能、迪美等企业等纷纷发现了防己在敏感皮肤上具有一定的舒敏效果,上海紫邦等也对粉防己碱和甘草酸二钾进行了复配增效,但是其存在活性物单一(多以粉防己碱为主,且上海紫邦专利中粉防己碱为辅助,占比低于5%,主要活性物还是甘草酸二钾),功效单一(局限于舒敏功效),药材利用度不高,萃取设备和工艺复杂、制备成本高的问题,此外,由于粉防己生物碱不溶于水,溶解度小,极性较小,logP达到7,穿过皮肤屏障能力弱,为达到良好效果,需要加大活性物质添加量,这就造成了成本高、皮肤刺激、过敏隐患增大等问题。
发明内容
针对上述不足,本发明的目的是提供一种具有祛痘消肿,抑菌,舒缓舒敏,去痘印疤痕等作用的含植物提取物的祛痘组合物。
一种含植物提取物的祛痘组合物,由以下重量份的组分制成:粉防己提取物60-90份,皮肤软化剂1-20份;黄柏提取物5-30份;龙胆提取物5-20份。
进一步的,上述的一种含植物提取物的祛痘组合物,所述的皮肤软化剂为1,3丙二醇、卡必醇、氮酮、戊二醇的其中之一。
进一步的,上述的一种含植物提取物的祛痘组合物,所述的皮肤软化剂由1,3丙二醇和卡必醇按照质量比25-50:1-10。
进一步的,上所述的含植物提取物的祛痘组合物,所述的粉防己提取物通过下述方法制备的:
1)将防己根茎切段后粉碎至40-60目粉末,以70%(v/v)乙醇为萃取溶剂,料液比1:10,萃取次数3次,每次1h,过300目滤网,合并滤液,得到提取液。
2)将提取液过陶瓷膜,收集滤过液通过纳滤膜,收集滤过液经反渗透膜脱盐,收集浓缩液经活性炭脱色,趁热过滤,去除活性炭,将滤液浓缩至浸膏,得到粉防己总生物碱提取物。
进一步的,上述的含植物提取物的祛痘组合物,所述的陶瓷膜的孔径为20-100nm。
进一步的,上述的含植物提取物的祛痘组合物,所述的纳滤膜孔径为700-1000Da。
上述的含植物提取物的祛痘组合物作为化妆品添加剂的应用;所述的含植物提取物添加为0.1-10%。
与现有技术相比,本发明提供的技术方案具有如下技术优点:
1)本发明提供的技术方案组方采用简易组合,精准分工,协同起效,采用中医药组方设计优选,以防己为君药,黄柏为臣药,龙胆为佐药,皮肤软化剂为使药,防己为君,苦寒止痛,辛散消肿,黄柏为臣,清热燥湿,控油收敛,龙胆为佐,修复痘痘创口,维稳修复,皮肤软化剂为使,软化皮肤屏障,直达毛囊深处。
2)本发明提供的技术方案药对协同,强化效果 防己与黄柏具有协同增效效果,能够强化消肿止痛效果。
3) 本发明提供的技术方案以皮肤软化剂强化经皮渗透,软化角质,增强效果,减少闭口导致的白头黑头问题。
4)本发明提供的技术方案温和、安全,适用于常规皮肤和敏感肌皮肤,经过了体外鸡胚测试和人体斑贴测试。
5)本发明提供的技术方案防己提取物采用陶瓷膜+纳滤膜+反渗透膜组合,采用物理截留方案,节能,高效,省时;针对性富集目标成分,工艺设计针对性强,提取物内组合物富含汉防己甲素,汉防己乙素等等多种成分,本发明提供的技术方案丙二醇体系承载力强,无活性物析出问题,且具有防腐效果,无需添加耐低温的防腐剂;采用1,3丙二醇、卡必醇、氮酮和戊二醇作为促渗剂组合物,能够针对性强化活性物经皮吸收,以较少用量及成本达到较好效果。
附图说明
图1是粉防己提取物HPLC色谱图;峰a 防己诺林碱;峰b 粉防己碱
图2是龙胆提取物B线性方程关系图;
实施方式
下面结合具体实施例对本发明的技术方案作进一步的详细说明,但不构成对本发明的任何限制,任何人在本发明权利要求保护范围内所做的非实质性改变,仍在本发明的权利要求保护范围内。
实施例1
本发明提供的一种含植物提取物的祛痘组合物,由下述重量份的组分组成:粉防己提取物70份,1,3丙二醇7.5份,卡必醇2.5份 ,黄柏提取物10份,龙胆提取物10份。
实施例2
本发明提供的一种含植物提取物的祛痘组合物,由下述重量份的组分组成:粉防己提取物60份,1,3丙二醇9份,卡必醇1份,黄柏提取物5份,龙胆提取物15份。
实施例3
本发明提供的一种含植物提取物的祛痘组合物,由下述重量份的组分组成:粉防己提取物80份,1,3丙二醇2.5份、卡必醇1份、氮酮1份、戊二醇1份 ,黄柏提取物15份,龙胆提取物5份。
实施例4
本发明提供的一种含植物提取物的祛痘组合物,由下述重量份的组分组成:粉防己提取物89份,1,3丙二醇5份 、卡必醇1份 、氮酮1份 ,黄柏提取物5份,龙胆提取物5份。
实施例5
一种含植物提取物的祛痘乳液,具体包括下述组分及其含量:丙烯酸(酯)类/C10-30烷醇丙烯酸酯交联聚合物0.2%,丁二醇5%,1,3-丙二醇3%,甘油2%,异壬酸异壬酯1%,辛酸/癸酸甘油三酯0.01%,甲基葡糖倍半硬脂酸酯1%,水溶性硅弹体5%,实施例1提供的祛痘组合物5%,氢氧化钠0.2%,剩余为去离子水。
制备工艺:
1)乳化锅中加入水,开搅拌形成漩涡后,加入丙烯酸(酯)类/C10-30烷醇丙烯酸酯交联聚合物,均质分散均匀后,加入丁二醇、1,3-丙二醇、甘油,搅拌加热至83℃,保温13min,
2)油相锅中加入异壬酸异壬酯,辛酸/癸酸甘油三酯,甲基葡糖倍半硬脂酸酯,开搅拌,升温到83℃,保温12min,溶解完全。
3)乳化锅中依次过滤加入步骤1)水相料和油相料,开搅拌,抽真空,均质乳化后,降温。
4)降温至48℃,加入氢氧化钠、水溶性硅弹体、实施例1提供的祛痘组合物,耐低温的防腐剂,搅拌12min,混合均匀,即可过滤出料。
实施例6
一种含植物提取物的祛痘乳液,具体包括下述组分及其含量:丙烯酸(酯)类/C10-30烷醇丙烯酸酯交联聚合物0.2%,丁二醇3%,1,3-丙二醇2%,甘油3%,异壬酸异壬酯2%,辛酸/癸酸甘油三酯1%,PEG-20-甲基葡糖倍半硬脂酸酯2%,肤感调节剂蓝丝美肤剂5%,异壬酯1%,辛酸/癸酸甘油三酯0.01%,甲基葡糖倍半硬脂酸酯1%,水溶性硅弹体5%,实施例2提供的祛痘组合物2%,氢氧化钠0.02%,耐低温的防腐剂0.5%,剩余为去离子水。
制备工艺:
1)乳化锅中加入水,开搅拌形成漩涡后,加入丙烯酸(酯)类/C10-30烷醇丙烯酸酯交联聚合物,均质分散均匀后,加入丁二醇、1,3-丙二醇、甘油,搅拌加热至80℃,保温15min。
2)油相锅中加入异壬酸异壬酯,辛酸/癸酸甘油三酯,乳化增稠剂,开搅拌,升温到85℃,保温10min,溶解完全。
3)乳化锅中依次过滤加入水相料和油相料,开搅拌,抽真空,均质乳化后,降温。
4)降温至50℃,加入氢氧化钠、肤感调节剂蓝丝美肤剂、祛痘组合物,耐低温的防腐剂,搅拌10min,混合均匀,即可过滤出料。
实施例7
一种含植物提取物的祛痘精华,具体包括下述组分及其含量:丙烯酸(酯)类/C10-30烷醇丙烯酸酯交联聚合物0.1%,丁二醇3%,1,3-丙二醇2%,甘油3%,实施例3提供的祛痘组合物15%,辛酸/癸酸甘油三酯0.001-5%,PEG-20-甲基葡糖倍半硬酯酸酯2%,肤感调节剂生物多糖胶5%,氢氧化钠0.01-0.5%,剩余为去离子水。
制备工艺:
乳化锅中加入水,开搅拌形成漩涡后,加入丙烯酸(酯)类/C10-30烷醇丙烯酸酯交联聚合物,均质分散均匀后,加入丁二醇、1,3-丙二醇、甘油,搅拌加热至82℃,保温13min,待溶解完全,加入乳化增稠剂、辛酸/癸酸甘油三酯,均质乳化后,降温至48℃,加入氢氧化钠、肤感调节剂、祛痘组合物,耐低温的防腐剂,搅拌13min,混合均匀,即可过滤出料。
实施例8
一种含植物提取物的祛痘膏霜,具体包括下述组分及其含量:卡波姆0.5%,丁二醇3%,1,3-丙二醇2%,甘油5%,异壬酸异壬酯1%,鲸蜡硬脂醇2%,辛酸/癸酸甘油三酯0.1%,乳化增稠剂PEG-20-甲基葡糖倍半硬酯酸酯0.5%,实施例3提供的祛痘组合物3%,肤感调节剂蓝丝美肤剂1%,氢氧化钠0.2%,剩余为去离子水。
制备工艺:
1)水相锅加入水,开搅拌形成漩涡后,加入卡波姆,搅拌分散均匀后,加入丁二醇、1,3-丙二醇、甘油,搅拌升温至81℃,保温14min溶解完全。
2)油相锅中加入异壬酸异壬酯,鲸蜡硬脂醇,辛酸/癸酸甘油三酯,乳化增稠剂PEG-20-甲基葡糖倍半硬酯酸酯,开搅拌,升温到83℃,保温12min溶解完全。
3)乳化锅中依次过滤加入水相料和油相料,开搅拌,抽真空,均质乳化后,降温。
4)降温至48℃,加入氢氧化钠、肤感调节剂蓝丝美肤剂、祛痘组合物,耐低温的防腐剂,搅拌12min,混合均匀,即可过滤出料。
对比例1
粉防己提取物70份,黄柏提取物10份,龙胆提取物10份。
对比例2
粉防己提取物70份,皮肤软化剂10份,黄柏提取物10份。
对比例3
皮肤软化剂10份,黄柏提取物10份,龙胆提取物10份。
对比例4
粉防己提取物70份,皮肤软化剂10份,龙胆提取物10份。
为了证明本发明提供的技术方案的优点,下面给出本申请提供的技术方案的的实验数据:
1、粉防己提取物提取物检测
粉防己提取物依照chp-2020-防己检测方法,流动相:甲醇:乙腈:水:冰乙酸=40:30:30:1;色谱柱:岛津 C18 柱(4.6×150mm,5μm);检测波长:282 nm;进行检测分析,以粉防己碱为标准品进行分析,粉防己提取物HPLC色谱图参阅图1;标准品及其他测试样品出峰参阅表1;
表1 标准品及样品出峰
序号 | 样品 | 粉防己碱浓度(mg/mL) | 防己诺林碱(mg/mL) |
1 | 粉防己提取物 | >20 | >10 |
2 | 实施例1 | 4-6 | 2-3 |
3 | 广州佳航-防己提取液(某市售提取液) | 0.01 | 未检出 |
4 | E-087(采购自道奇) | 0.00 | 0.00 |
2、防腐挑战测试
防腐挑战原理:通过在待测样本中人为地接种若干种类、一定数量的微生物,在适当的温度下存放,定期分离样本的微生物,并根据微生物的数量变化情况评价样本的防腐性能。
防腐挑战测试方法:通过在产品中分别接入一定数量的细菌混合物和真菌混合物(分为有大肠杆菌、金黄色葡萄球菌、铜绿假单胞菌混合物1×108CFU/mL-9.0×108CFU/mL和白色念珠菌、黑曲霉混合物1.0×107CFU/mL-9.0×107CFU/mL),并在第7天、14天、28天检测产品中微生物含量,通过内部检测评判标准评判产品的防腐性能。
测试通过判断标准 (对数减少值)
细菌: 7d:≥3,14d:≥3 且NIb,28d:>5
真菌: 7d:≥2,14d:≥2 且NIb,28d:>4
防腐挑战测试数据(细菌)—:表示未检测;
防腐挑战测试数据(真菌)“—:表示未检测
微生物检测评判标准和实施案例1防腐测评结果如表2所示。
表2 细菌测试结果
表3 真菌测试结果
结果显示:由测试结果显示实施案例1产品通过防腐挑战实验,而对比例1在进行防腐挑战测试中在第二周菌落数量开始逐渐增多,在第四周测试时菌落已经超过测试标准,防腐挑战测试不通过。
3、稳定性测试
稳定性测试原理:通过将待测样品放入高温、低温、冷热交替的环境中,观测待测样品状态、颜色等变化来评估样品的稳定性。
稳定性测试项目主要包括耐热45±1℃、耐寒-5℃、冷热交替循环测试48±1℃维持24h,-20±1℃维持24h,交替循环,第7、14、21、30、60、90天分别进行样品观测,结果见下表4所示。
表 4 稳定性测试结果
结果分析:实施例1提供的组合物在测试周期内状态稳定,色泽清亮,澄清透明,而对比例1提供的组合物在测试过程分别出现析出颗粒物,体系浑浊,微生物超标等系列问题,表明对比例1体系承载性好,且具有防腐效果,丙二醇在适当比例下具有良好的溶解效果。
5、透皮促渗透效果测试
以经典FRANZ扩散池法进行体外透皮实验,选用1月龄乳猪皮为测试皮肤,进行体外透皮测试,以粉防己碱为标准品进行构建标准曲线进行检测分析,体外实验如下:
将实施例1和对比例1均转化为乳液(5%添加),以FRANZ立式扩散池为试验平台,乳猪皮角质层朝上,均匀涂抹在角质层上,以生理盐水作为接收液,设置测试温度32℃,磁力搅拌转速350rpm,连续透皮测试8h(参考化妆品使用时长),进行试验,平行6组,计算累计透皮量,结果如表5所示。
表5 透皮促渗透效果测试结果
结果表明,相对于对比例1(无促渗透剂的组合物),实施例1(有促渗透剂组合物)的活性物粉防己碱皮肤渗透效果显著优于对比例,其活性物穿过皮肤角质层,到达皮下组织的量显著高于对比例。
6、祛痘抑菌功能效果
方法原理:以经典牛津杯抑菌圈方法进行痤疮丙酸杆菌抑制实验
方法简述:无菌操作下,取106~107 CFU/mL的痤疮丙酸杆菌混合液于培养基中。并将牛津杯放入凝固的培养基表面,在牛津杯加入受试物200ul和空白对照组生理盐水200ul,然后进行厌氧培养,温度设置为36±1℃,恒温培养24-72小时后取出,用工具尺测量出抑菌圈直径的大小。每个样品设置三个平行。待测样品抑菌检测结果如表6所示。
表6 祛痘抑菌功能效果结果
结果分析:实施例1-4对痤疮丙酸杆菌抑制效果较为明显,抑菌圈直径达到20mm以上,对比例1效果最弱,基本没有抑菌效果,对比例2抑菌圈达到17.50mm,对比例3和对比例4效果较差。表明组方中主要抑菌成分为黄柏提取物+防己提取物+皮肤软化剂,且具有良好的增效效果。
7、巨噬细胞抗炎测试(TNF-a抑制)
实验原理简述:通过LPS诱导巨噬细胞产生炎症因子TNF-a构建细胞分泌炎症因子模型,通过受试物与空白对照组处理细胞后,检测并比对分析TNF-a的含量;通过TNF-a的含量计算TNF-a分泌抑制效率,验证受试物的抑制炎症因子分泌效果。
将RAW264.7小鼠巨噬细胞调整为1*105个/mL浓度并接种于6孔板内,每孔2mL,置于细胞培养箱中培养24h。弃去原培养基后分别加入2mL完全培养基,含有5ug/mL LPS的完全培养基2mL,含有5ug/mL LPS且含有质量分数分别为10%样品的完全培养基2mL继续培养24h。搜集细胞上清液,由ELISA试剂盒说明书进行TNF-a含量检测。每组设立三个复孔。
样品说明:因细胞实验对角质软化剂特别敏感,因此均不添加角质软化剂测试,组合物中角质软化剂以纯水替代。测试样品为1%样品稀释测试,稀释液为细胞培养液上清液,测试结果表7所示。
表7 TNF-a分泌抑制效率
序号 | 样品 | TNF-a分泌抑制效率(Mean±SD) |
1 | 实施例1 | 88.33%±11.28 |
2 | 实施例2 | 82.24%±11.27 |
3 | 实施例3 | 88.47%±11.58 |
4 | 实施例4 | 92.64%±12.19 |
5 | 对比例1 | 84.22%±10.25 |
6 | 对比例2 | 64.23%±11.34 |
7 | 对比例3 | 65.28%±13.36 |
8 | 对比例4 | 78.26%±11.28 |
结果分析:巨噬细胞抗炎试验结果显示,实施例样品对TNF-a具有良好的抑制分泌效果,对比例1 与实施例差异不大,而对比例2-4显著低于实施例,可能是由于龙胆和粉防己缺失后无法形成协同抑制效果,对比例4缺乏黄柏提取物,但是效果也达到了78.26%,与实施例差异不明显,表明组方中主要的抗炎物质为龙胆和防己,黄柏为辅助抗炎物质。
8、角质细胞增殖
实验原理简述:角质细胞是对皮肤屏障具有决定影响的细胞,痤疮发生过程中多会伴随毛孔堵塞产生的粉刺、白头等,进一步会因挤压、撕扯导致屏障受损,因此在局部屏障受损后需要及时进行角质细胞增殖修复屏障,因此角质细胞增殖试验具有一定的指示。
首先通过MTT检测法,检测受试物对角质形成细胞活力影响。进一步将角质细胞与活性物进行共培养观察细胞增加量。
因细胞实验对角质软化剂特别敏感,因此均不添加角质软化剂测试,组合物中角质软化剂以纯水替代。测试样品为1%样品稀释测试,稀释液为细胞培养液上清液;实验结果如8所示。
表8 角质形成细胞增殖率
序号 | 样品 | 角质形成细胞增殖率 |
1 | 实施例1 | 112.33%±12.28 |
2 | 对比例2 | 87.23%±13.34 |
3 | 对比例3 | 106.28%±10.36 |
4 | 对比例4 | 98.26%±14.26 |
5 | 阴性组 | 100% |
结果分析:角质细胞增殖试验以阴性对照组为100%,进行测试分析发现实施例1具有良好的细胞增殖效果,增值率达到112.33%,对比例2对细胞具有显著抑制效果,表明黄柏和防己可能产生一定的细胞毒性,对比例3对细胞具有少许增殖效果,表明防己可能是主要的细胞毒性物质,对比例4与空白组相比没有增殖效果。
9、 DPPH抗氧化效果
测试原理:当人体产生的过量自由基能,会导致各种肌肤问题。通过体外DPPH自由基抑制实验检测受试物清除DPPH效果。
DPPH是一种稳定的以氮为中心的自由基,在519nm波长处有最大吸收值。DPPH的乙醇溶液呈深紫色,当自由基清除剂加入到DPPH溶液中时,DPPH的单电子被配对而使其颜色变浅,在最大吸收波长处的吸光值随之减小。因此,可通过吸光值减弱的程度来判断物质清除DPPH自由基的能力
将DPPH用无水乙醇配制成65.7mg/L的DPPH溶液工作液。取2mL待测样品与4mLDPPH溶液工作液混合,常温避光反应30min后于紫外分光光度计517nm波长检测吸光值。DPPH•的清除率(%)=[1-(B-C)/A]*100%(A:对照液(无水乙醇代替待测样品)测定的吸光值;B:受试物测定的吸光值;C:试样空白(无水乙醇液代替DPPH)吸光值 )。各组案例DPPH自由基清除率效果如表9所示。
表9 DPPH自由基清除率%
结果分析 实施例1-4均表现出练好的清除自由基效果,自由基清除率均达到88%以上,实施例4效果最优,达到92.35%,对比例1抑制效果不佳,可能因其缺乏丙二醇增溶,粉防己溶解不佳效,对比例3基本没有效果,推测是粉防己提取物含量不足。
10、 成纤维细胞培养抑制及胶原蛋白分泌抑制试验
实验原理:皮肤在受到创伤后的愈合过程依赖于成纤维细胞分泌胶原蛋白,但是人为地挤压痘痘造成的皮肤创口、烧伤、割伤等时候,由于机体调节和免疫作用,会导致皮肤内部成纤维细胞过度分生,胶原蛋白过量分泌,且不规则分布交联,导致了瘢痕、痘坑、痘印的产生,通过选取增生性瘢痕患者组织的病理细胞,通过体外细胞培养抑制试验和体外细胞胶原蛋白分泌抑制试验能够预判痘坑,痘印等皮肤问题的产生概率。设置实验组和对照组,对照组不加样品,实验组进行MTT试验确定细胞毒性试验后选择合适的试验浓度,加入测试样品与细胞共培养,比较对照组和实验组的细胞存活及胶原蛋白分泌数据;测试结果如表10所示。
细胞培养抑制率=(对照组-实验组)÷对照组×100%
胶原蛋白I 分泌抑制率%(对照组-实验组)÷对照组×100%
表10 成纤维细胞培养抑制及胶原蛋白分泌抑制
序号 | 样品 | 细胞抑制率%(Mean±SD) | 胶原蛋白I 分泌抑制率%(Mean±SD) |
1 | 实施例1 | 60.81%±13.29 | 58.81%±15.35 |
2 | 实施例2 | 75.28%±11.28 | 65.28%±14.20 |
3 | 实施例3 | 72.35%±12.29 | 62.35%±13.49 |
4 | 实施例4 | 74.48%±13.06 | 64.48%±11.26 |
5 | 对比例2 | 94.90%±10.20 | 82.90%±9.29 |
6 | 对比例3 | 91.38%±1.29 | 91.25%±3.27 |
结果分析 实施例1-4均表现出对抑制成纤维细胞增殖分生效果,同步也一致胶原蛋白分泌,对比例对成纤维细胞的抑制效果基本不明显, 表明粉防己提取物可能具有减少成纤维细胞在痘印,痘坑形成方面的效果
11、兔耳痤疮试验模型
实验原理:对新西兰大白兔的双耳外耳道进行兔耳痤疮造模,通过用0.25 mL注射器均匀涂布 50%油酸 0.2 mL, 1 次/d, 连续 28 d。第12天在双侧兔耳皮内注射表皮葡萄球菌30 μL。对造模成功后的进行受试物处理,检测给药后的兔耳的皮脂腺直径和毛囊面积大小来评估样品的祛痘效果(P<0.05)。
实验方法:确认造模成功后将其分成四组, 即 B 组:模型组; C 组:实验组;D组:对照组(复方珍珠暗疮片),同时设立正常组A组,每组设置8个平行,连续涂抹14天后通过组织免疫检测皮脂腺直径、毛囊面积。测试结果如表11所示。
表11 兔耳痤疮试验模型
结果分析:经兔耳朵试验造模,模型组相对于正常组皮脂腺直径和毛囊面积显著增大,阳性对照组采用复方珍珠暗疮片进行治疗,其皮脂腺直径和毛囊面积显著低于模型组,表明模型造模成功,各实验组数据展现较大差异,实施例1相对于模型组达到极显著水平P<0.01,与阳性对照组相同,效果明显,对比例1、对比例3、对比例4无明显差异,对比例2相对于模型组具有显著效果,鉴于对比例2含有粉防己和黄柏提取物,其可能是达到祛痘效果的主要物质。
12、 安全性测试
1、利用孵化10-14天的受精鸡胚中期绒毛尿囊膜(CAM)的血管结构和人眼结膜血管组成较为相似的特征原理,利用鸡胚测试方法检测样品的眼刺激性。将40 ul受试物直接与鸡胚尿囊膜接触,同时设置阴性对照组(生理盐水)和阳性对照组(0.01mol/LSDS),通过作用30 min前后的图片进行对比,观察绒毛尿囊膜血管损伤(如鬼影血管、毛细血管充血或出血)程度,对不同的血管损伤程度进行评分,从而判断受试物的刺激性大小。
NC值:根据血管损伤程度评分的平均分大小(即NC)值来比较受试物的刺激性大小,评价标准如表12所示;测试结果如表13所示。
表12 NC值评价标准
分值 | 评价 | 表述 |
0<NC≤1.64 | 无刺激 | 无损伤性刺激,此时毛细血管未有出血损伤 |
1.64<NC≤3 | 轻微刺激 | 轻微损伤性刺激,此时毛细血管出现轻微性出血损伤 |
3<NC≤5 | 中度刺激 | 较为严重的损伤性刺激,此时毛细血管出现较为严重的出血损伤,可能为不可逆损伤 |
NC>5 | 严重刺激 | 较为严重的损伤性刺激,此时毛细血管大面积出血,且出血量较大,为不可逆损伤 |
表13 刺激测试结果
2、斑贴实验
采用实施例5提供的产品进行斑贴试验。参照《化妆品安全技术规范》、国国家标准GB17149.1-1997《化妆品皮肤病诊断标准及处理原则》、国国家标准GB17149.2-1997《化妆品接触性皮炎诊断标准及处理原则》进行人体斑贴试验。本研究招募志愿者24名,并签署志愿者知情同意书,先将所有斑试器背面做样品序号标记并加入等量待测样品,对照组为不加任何物质的空白组,将处理好的斑试器贴敷于受试者的前臂曲侧,设置三组平行和一个空白组,分别持续24h、48h后观察皮肤状态。样品测试结果如表14所示,使用实施例5提供的产品的30例受试者均为阴性反应;对人体无不良反应。
表 14 斑贴实验结果
13、人体测试
选取面部有明显红、肿、凸起等痤疮状况和闭口粉刺的志愿者(依据Pillsbury四级改良分级法,选定1级症状)60名,年龄区间为20 ~ 30 岁,并签志愿者知情同意书。将志愿者随机分为两组,每组30人,一组志愿者使用空白膏基组,另外一组志愿者使用实施例5提供的产品,产品使用3天后,通过消费者使用问卷调查形式。志愿者分别对痘痘红肿减轻、痘痘红肿消退、痘痘疼痛减轻、痘痘疼痛消失、痘痘部分消失、痘痘大部分消失等维度进行评分。评价结果如下表15所示。
表15 消费者祛痘评价
试用产品 | 痊愈 | 显效 | 有效 | 无效 | 总有效率(%) |
空白膏基组 | 1 | 2 | 4 | 23 | 23.33% |
实施例5 | 6 | 9 | 10 | 5 | 83.33% |
由测试结果显示,实施例5提供的产品对痘痘肌肤具有较好的治愈效果,总有效率达到 83.33%,痊愈认识,显效人数,有效人数均优于空白基质,空白基质组基本没有显著治愈效果。
Claims (7)
1.一种含植物提取物的祛痘组合物,其特征在于,由以下重量份的组分制成:粉防己提取物60-90份,皮肤软化剂1-20份,黄柏提取物5-30份,龙胆提取物5-20份;
所述的皮肤软化剂为1,3丙二醇、卡必醇、氮酮、戊二醇的至少一种。
2.根据权利要求1所述的一种含植物提取物的祛痘组合物,其特征在于,所述的皮肤软化剂由1,3丙二醇和卡必醇按照质量比25-50:1-10组成。
3.根据权利要求1所述的含植物提取物的祛痘组合物,其特征在于,所述的粉防己提取物通过下述方法制备:
1)将防己根茎切段后粉碎至40-60目粉末,以70%乙醇为萃取溶剂,料液比1:10,萃取次数3次,每次1h,过300目滤网,合并滤液,得到提取液;
2)将提取液过陶瓷膜,收集滤过液通过纳滤膜,收集滤过液经反渗透膜脱盐,收集浓缩液经活性炭脱色,趁热过滤,去除活性炭,将滤液浓缩至浸膏,得到粉防己总生物碱提取物。
4.根据权利要求3所述的含植物提取物的祛痘组合物,其特征在于,所述的陶瓷膜的孔径为20-100nm。
5.根据权利要求3所述的含植物提取物的祛痘组合物,其特征在于,所述的纳滤膜孔径为700-1000Da。
6.权利要求1-5任一所述的含植物提取物的祛痘组合物作为制备化妆品添加剂的应用。
7.权利要求1-5任一所述的含植物提取物的祛痘组合物作为制备化妆品添加剂的应用,所述的含植物提取物添加为0.1-10%。
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