CN114469826B - 一种凝胶型浙麦冬美白面膜及其制备方法 - Google Patents
一种凝胶型浙麦冬美白面膜及其制备方法 Download PDFInfo
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- CN114469826B CN114469826B CN202210081172.0A CN202210081172A CN114469826B CN 114469826 B CN114469826 B CN 114469826B CN 202210081172 A CN202210081172 A CN 202210081172A CN 114469826 B CN114469826 B CN 114469826B
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- radix ophiopogonis
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Abstract
本发明公开了一种凝胶型浙麦冬美白面膜及其制备方法，包括以下重量百分比的组分：麦冬高异黄酮提取物1‑6%、吸收促进剂0.05‑0.5%、络合剂1‑7%、美白增强剂0.04‑0.4%、卡波姆980 0.2‑0.8%、透明质酸钠0.01‑0.05%、甘油10‑20%、乙二醇1‑5%、茶油0.05‑0.1%、吐温‑80 0.05‑2%、氮酮0.05‑2%、尼泊金乙酯0.1‑0.5%、尿囊素0.05‑0.5%、余量为去离子水。本发明配方无毒、无刺激、安全性能高，且浙麦冬高异黄酮提取物能有效抑制酪氨酸酶活性，有效减轻黄褐斑及痤疮引起的黑斑且来源丰富，且属于可再生资源，绿色环保。
Description
技术领域
本发明属于化妆品技术领域，具体涉及一种凝胶型浙麦冬美白面膜及其制备方法。
背景技术
面膜是集清洁、护理、营养于一体的面部皮肤用化妆品，深受成年人尤其是女性消费者喜爱。随着消费心理的成熟，人们不仅要求面膜可以达到清洁、护理、营养的基本功效，还进一步追求开发面膜的美白、抗衰老等高级功效。目前，面膜的种类繁多，分为贴式面膜、撕拉面膜、凝胶面膜等。其中凝胶型面膜为凝胶状膏体，易于均匀涂抹在面部，可以阻隔面部皮肤与空气的接触，有效防止水分的蒸发，增加角质层的湿度，软化角质；还能使皮肤表面毛孔扩张，有利于营养成分吸收。
皮肤颜色的深浅受多种因素调控，主要取决于黑色素的含量及分布。黑色素是一种氨基酸衍生物，是酪氨酸或3,4-二羟苯丙氨酸经过一连串化学反应所形成的，其常以聚合的方式存在。黑色素由人表皮基底层中的黑色素细胞合成，当紫外线照射到皮肤上，肌肤就会处于“自我防护”的状态，激活酪氨酸酶的活性，催化酪氨酸或3,4-二羟苯丙氨酸氧化生成黑色素，来保护我们的皮肤细胞。黑色素又经由细胞代谢的层层移动，到了肌肤表皮层形成雀斑、晒斑、黑斑等。
随着近年来对中药药效活性物质的研究不断深入与技术现代化的发展，越来越多的中药被发现具有美容的功效而被运用到护肤品中。浙麦冬是浙江省最为著名的道地药材“浙八味”之一，用于急性咳嗽、喉咙肿痛和心脑血管等疾病的治疗已有上千年的历史。2002年，麦冬被国家卫健委批准为可用于保健食品的“药食同源”中药；此外，其在日本及越南等东南亚国家也常作为功能食品和地方药品使用。研究表明，黄酮类成分是其主要药效物质之一，在我们前期的研究中发现，浙麦冬块根及须根中均含有特征性黄酮类成分，包括甲基麦冬黄烷酮A，甲基麦冬黄烷酮B，6-醛基异麦冬黄烷酮A和6-醛基异麦冬黄烷酮B等均具有抑制酪氨酸酶的作用，从而阻断细胞内黑色素的生成，因此，可用于美白等作用的护肤品中，达到美容祛斑的作用。而此前麦冬提取物已被用于多个美白面膜制备的专利中，如中国专利CN111956543A公开了一种美白抗敏的大麻叶面膜，所述中药组合物由以下重量份的原料组成：水60-92％、丙二醇1-6％、甘油1-6％、扭刺仙人掌茎提取物0.05-3％、麦冬根提取物0.05-3％、苦参根提取物0.05-3％、海藻糖0.1-6％、丁二醇0.1-6％、雪莲花提取物0.05-3％、高山火绒草花/叶提取物0.05-3％、酒石酸0.001-0.3％、EDTA二钠0.001-0.3％、亚硫酸钠0.001-0.3％、焦亚硫酸钠0.001-0.3％、银耳提取物0.1-2％、对羟基苯乙酮0.1-1％、1,2-己二醇0.1-1％、黄原胶0.05-0.2％、尿囊素0.05-0.2％、大麻叶提取物0.05-5％、聚丙烯酸钠0.05-0.2％。制备的面膜中含有0.05-3％麦冬根提取物。中国专利CN113041205A一种美白保湿中药面膜及其制备方法，以菟丝子10-300重量份、红花10-200重量份、白蒺藜10-100重量份、麦冬1-50重量份、韭菜叶50-500重量份、黄土5-50重量份、植物油10-100重量份、淀粉100-1000重量份作为原料，经过粉碎、超声提取、调节pH等步骤,制得一种美白保湿中药面膜。尽管以上面膜制备方法所述的配方中含有麦冬提取物，但是从配方组成看均不是以麦冬提取物为主药，而且并未明确面膜中麦冬功效成分的结构类型及其含量标准。因而，基于我们已有研究实验结果，开发一种含有浙麦冬高异黄酮提取物的中药面膜将具有很好的美白功效和市场潜力。
发明内容
针对现有技术中存在的问题，本发明设计的目的在于提供一种凝胶型浙麦冬美白面膜及其制备方法，具体通过以下技术方案加以实现：
所述的一种凝胶型浙麦冬美白面膜，包括以下重量百分比的组分：浙麦冬高异黄酮提取物1-6％、吸收促进剂0.05-0.5％、络合剂1-7％、美白增强剂0.04-0.4％、卡波姆9800.2-0.8％、透明质酸钠0.01-0.05％、甘油10-20％、乙二醇1-5％、茶油0.05-0.1％、吐温-80 0.05-2％、氮酮0.05-2％、尼泊金乙酯0.1-0.5％、尿囊素0.05-0.5％、余量为去离子水。
进一步地，浙麦冬高异黄酮提取物6％、吸收促进剂0.4％、络合剂1％、美白增强剂0.4％、卡波姆980 0.2％、透明质酸钠0.01％、甘油18％、乙二醇1％、茶油0.05％、吐温-802％、氮酮2％、尼泊金乙酯0.1％、尿囊素0.05％、余量为去离子水。其中美白增强剂中维生素B1 0.2％，微生物C 0.2％，吸收促进剂中浙麦冬甾体皂苷提取物0.3％，甘氨胆酸钠0.1％。
进一步地，浙麦冬高异黄酮提取物是指：浙麦冬高异黄酮提取物按干燥品计，所含甲基麦冬黄烷酮A，甲基麦冬黄烷酮B，6-醛基异麦冬黄烷酮A和6-醛基异麦冬黄烷酮B的总量不得少于10％。
进一步地，吸收促进剂由浙麦冬甾体皂苷提取物和胆酸盐组成；所述浙麦冬甾体皂苷与胆酸盐的质量比为3:1；浙麦冬甾体皂苷提取物按干燥品计算，含麦冬总皂苷以鲁斯可苷元计，不得少于5％，胆酸盐选用甘氨胆酸钠、甘氨鹅脱氧胆酸钠、牛磺胆酸钠、牛磺鹅脱氧胆酸钠、脱氧胆酸钠中的一种或两种以上。
进一步地，所述络合剂为EDTA-二钠或壳聚糖中的一种或二种，EDTA-二钠的重量百分比为2-3％，壳聚糖的重量百分比为1-4％。
进一步地，美白增强剂为维生素B1和维生素C中的一种或二种。
进一步地，茶油为冷榨山茶油，山茶油媲美橄榄油，脂肪酸组成与橄榄油相似。制备包括以下步骤：
选取成熟山茶籽，40-50℃烘干干燥至含水量3-6％，干燥后的山茶籽送入脱壳机脱壳；将脱壳后的山茶籽冷榨压榨出山茶油，榨膛温度为100～150℃，出油温度为80～120℃；压榨出的山茶油通过板框压滤机除去固体杂质；将板框压滤后的山茶油于-5℃～5℃冬化；冬化后的山茶油经过分子蒸馏除去山茶油中的游离脂肪酸、油脂过氧化产物、水分及其他低沸点物质；分子蒸馏后的山茶油用白土、活性炭和氧化铝混合物作为过滤介质过滤除去杂质和色素即得成品。
进一步地，浙麦冬高异黄酮提取物的制备方法为：
1)浙麦冬粉末加入10-20倍量的70％的乙醇，80℃加热回流提取1-2h，提取1-3次，滤液浓缩得到浙麦冬提取物浸膏，加去离子水稀释成浓度至每1ml相当于原有药材0.1-1g的水提取物；
2)取步骤1)的提取物，按提取物与树脂的固液比为1-10mL/g上样于D101大孔树脂，依次用蒸馏水、60％的乙醇、90％乙醇洗脱，收集90％乙醇洗脱液，减压浓缩，真空冷冻干燥得到提取物1，即浙麦冬高异黄酮提取物；其中，收集60％乙醇洗脱液，减压浓缩，真空冷冻干燥得到提取物2，即浙麦冬甾体皂苷提取物。
所述的一种凝胶型浙麦冬美白面膜的制备方法，包括以下步骤：
1)A相的制备：将配方量的卡波姆980倒入适量去离子水中搅拌，待分散均匀后加入配方量的尿囊素和氮酮继续搅拌加热至80℃恒温15min；
2)B相的制备：在配方量的甘油中加入配方量的透明质酸钠搅拌，待透明质酸分散均匀后加入适量蒸馏水继续搅拌，待体系均匀后再加入配方量的茶油和乙二醇并加热搅拌至80℃恒温15min；
3)C相的制备：将配方量的浙麦冬高异黄酮提取物、促进吸收剂、美白增强剂加入适量的去离子水中，并滴加少量乙醇，温热至55℃并用超声波溶解；
4)将B相组分加入到A相组分中，加入配量的络合剂，于80℃搅拌15min后冷却至55℃，加入溶解均一的C相组分，并使产品继续冷却至45℃，然后依次加入配方量的尼泊金乙酯和吐温-80，加去离子水至100mL，搅拌均匀即得膏体；
5)将膏体置于真空搅拌机内，加入去离子水补足余量，真空搅拌均匀，包装即得凝胶型浙麦冬美白面膜。
进一步地，步骤1)中适量去离子水占总去离子水用量的70-80％，步骤4)中络合剂为EDTA-二钠和壳聚糖两者中的一种或两种。
一种凝胶型浙麦冬美白面膜在减轻黄褐斑及痤疮引起的黑斑上应用。
酪氨酸酶又称为多酚氧化酶，是生物体内黑色素生物合成过程中的关键限速酶。在黑色素形成的过程中，酪氨酸酶可催化L-酪氨酸生成L-多巴，L-多巴逐步氧化成多巴醌，进而形成色素颗粒，其过高的活性会引发色素沉积、黄褐斑、雀斑、老年斑等症状。因而酪氨酸酶抑制活性常被用于评价美白化妆品的祛斑美白功效，如曲酸、熊果苷等酪氨酸酶抑制剂已作为美白剂在化妆品、护肤品中得到广泛应用。
浙麦冬中的特征性成分高异黄酮是一类天然存在的独特黄酮类结构化合物，其母核结构比含两个芳香环的氧杂环化合物的异黄酮在B和C环之间多一个碳原子。对该类化合物的生物学研究表明，其具有多种生理功能，如强效抗真菌，抗氧化剂，抗增殖，抗炎，抗组胺，抗过敏，细胞毒性，抗诱变，和抗血管生成活性(如cremastranone)。特别地，该类化合物也具有蛋白质酪氨酸激酶(PTK)抑制活性。
本发明研究表明，浙麦冬4种高异黄酮单体成分及其混合提取物具有显著的抑制酪氨酸酶的活性，能有效减轻黄褐斑及痤疮引起的黑斑，具有祛斑美白、保湿的作用。同时本发明研究表明，麦冬水提物酪氨酸酶抑制活性弱，麦冬多糖不具有酪氨酸酶抑制活性，并且麦冬多糖粘性强，不利于用于面膜的制备。因此，采用麦冬高异黄酮提取物作为主要面膜中功能活性成分，具有酪氨酸酶抑制活性物质得到有效富集，效果和安全性更高。
本面膜配方中的其他的辅料成分是面膜中常用且经实验证明了安全性。如甘油、尿囊素具有保湿的功效，透明质酸钠本身是人体皮肤的构成之一，存在于结缔组织中，由于透明质酸钠含有大量的羧基和羟基，在水溶液中形成分子内和分子间的氢键，具有良好的保湿作用。壳聚糖和橙皮素能通过分子中的氨基、羟基络合面部的Pb2+、Hg2+、Ni2+等重金属离子，除去皮肤中的重金属离子，降低重金属的危害。
本发明与现有技术相比，具有以下有益效果：
1)本发明中使用的浙麦冬高异黄酮提取物能有效抑制酪氨酸酶活性，有效减轻黄褐斑及痤疮引起的黑斑。且提取物物中功能成分清晰，效果确切，过敏原筛查明确清晰。浙麦冬高异黄酮提取物从天然植物浙麦冬中提取制备而得，来源丰富，且属于可再生资源，绿色环保；
2)本发明配方在利用浙麦冬高异黄酮提取物作为功能因子时，以浙麦冬甾体皂苷提取物作为一种天然的非离子型表面活性剂，实现了浙麦冬资源的高效综合利用，利用皂苷的结构特性能够在水中形成微小的胶体粒子，通过形成胶束作用将配方中的活性成分吸附在一起，增加细胞间水溶性通道的渗透性，促进水溶性物质的吸收；并且低浓度的胆酸盐可以水化并抽提细胞间脂质，从而增加了提取物在细胞间水性通道内的扩散，同时胆酸盐可作为酶抑制剂，通过影响酶的活性，改变多肽、蛋白质构象等不同的机制维持药物的稳定，将其与表面活性剂浙麦冬甾体皂苷提取物复配能够明显增大美白面膜中有效成分的吸收；
3)本发明配方中的所有组分均为无毒、无刺激、安全性能高的成分，大部分成分为天然植物提取活性成分，可以放心使用。
附图说明
图1为高异黄酮标准品测定HPLC色谱图；
图2浙麦冬高异黄酮提取物中高异黄酮含量测定HPLC色谱图；
图中，1为甲基麦冬黄烷酮A；2为甲基麦冬黄烷酮B；3为6-醛基异麦冬黄烷酮A；4为6-醛基异麦冬黄烷酮B。
具体实施方式
以下结合具体实施例对本发明做进一步详细描述，以便更好地理解本技术方案，本发明中所涉及的百分比除非另有说明，均是以重量百分比计算。
实施例1
浙麦冬高异黄酮提取物3％、维生素C 0.3％、浙麦冬甾体皂苷提取物0.3％、甘氨胆酸钠0.1％、卡波姆980 0.5％、透明质酸钠0.02％、甘油15％、乙二醇3％、茶油0.1％、吐温-80 1％、氮酮1％、尼泊金乙酯0.3％、尿囊素0.3％、壳聚糖1％，余量为去离子水。
制备方法：
1)A相的制备:将0.5重量份卡波姆980倒入55重量份的去离子水中搅拌，待分散均匀后加入0.3重量份尿囊素和1重量份氮酮继续搅拌加热至80℃恒温15min；
2)B相的制备:在15重量份甘油中加入0.02重量份透明质酸钠搅拌，待透明质酸分散均匀后加入适量蒸馏水继续搅拌，待体系均匀后再加入0.1重量份茶油和3重量份乙二醇并加热搅拌至80℃恒温15min；
3)C相的制备:加入提取出来的3重量份浙麦冬高异黄酮提取物，0.3重量份浙麦冬甾体皂苷提取物，0.1重量份甘氨胆酸钠、0.3重量份维生素C和适量水，并滴加少量乙醇，温热至55℃并用超声波溶解；
4)将B相组分加入到A相组分中，加入1重量份壳聚糖，于80℃搅拌15min后冷却。待冷却至55℃，加入溶解均一的C相组分，并使产品继续冷却。待产品冷却至45℃加入0.3重量份尼泊金乙酯，最后加入1重量份吐温-80，加去离子水，搅拌均匀即得膏体；
5)将膏体置于真空搅拌机内，加入去离子水补足余量，真空搅拌均匀，包装即得。
实施例2
麦冬高异黄酮提取物5％、维生素C 0.2％、浙麦冬甾体皂苷提取物0.36％、甘氨胆酸钠0.12％、卡波姆980 0.6％、透明质酸钠0.04％、甘油20％、乙二醇4％、茶油0.1％、吐温-80 2％、氮酮2％、尼泊金乙酯0.4％、尿囊素0.4％，EDTA-二钠3％，余量为去离子水。
制备方法：
1)A相的制备:将0.6重量份卡波姆980倒入45重量份的去离子水中搅拌，待分散均匀后加入0.4重量份尿囊素和2重量份氮酮继续搅拌加热至80℃恒温15min；
2)B相的制备:在20重量份甘油中加入0.04重量份透明质酸钠搅拌，待透明质酸分散均匀后加入适量蒸馏水继续搅拌，待体系均匀后再加入0.1重量份茶油和4重量份乙二醇并加热搅拌至80℃恒温15min。
3)C相的制备:加入提取出来的5重量份浙麦冬高异黄酮提取物，0.4重量份浙麦冬甾体皂苷提取物，0.13重量份甘氨胆酸钠、0.2重量份维生素C和适量水、并滴加少量乙醇，温热至55℃并用超声波溶解。
4)将B相组分加入到A相组分中，加入3重量份EDTA-二钠，于80℃搅拌15min后冷却。待冷却至55℃，加入溶解均一的C相组分，并使产品继续冷却。待产品冷却至45℃加入0.4重量份尼泊金乙酯，最后加入2重量份吐温-80，加去离子水，搅拌均匀即得膏体；
5)将膏体置于真空搅拌机内，加入去离子水补足余量，真空搅拌均匀，包装即得。
实施例3
麦冬高异黄酮提取物2％、维生素B1 0.3％、浙麦冬甾体皂苷提取物0.21％、甘氨胆酸钠0.07％、卡波姆980 0.8％、透明质酸钠0.03％、甘油15％、乙二醇5％、茶油0.05％、吐温-80 1.5％、氮酮1.5％、尼泊金乙酯0.5％、尿囊素0.2％、EDTA-二钠2％，余量为去离子水。
制备方法：
1)A相的制备:将0.8重量份卡波姆980倒入50重量份去离子水中搅拌，待分散均匀后加入0.2重量份尿囊素和1.5重量份氮酮继续搅拌加热至80℃恒温15min；
2)B相的制备:在重量份甘油中加入0.03重量份透明质酸钠搅拌，待透明质酸分散均匀后加入适量蒸馏水继续搅拌，待体系均匀后再加入0.05重量份茶油和5重量份乙二醇并加热搅拌至80℃恒温15min；
3)C相的制备:加入提取出来的2重量份浙麦冬高异黄酮提取物，0.2重量份浙麦冬甾体皂苷提取物、0.06重量份甘氨胆酸钠，0.3重量份维生素B1和适量水，并滴加少量乙醇，温热至55℃并用超声波溶解；
4)将B相组分加入到A相组分中，加入2重量份EDTA-二钠2％，于80℃搅拌15min后冷却。待冷却至55℃，加入溶解均一的C相组分，并使产品继续冷却。待产品冷却至45℃加入0.5重量份尼泊金乙酯，最后加入1.5重量份吐温-80，加去离子水，搅拌均匀即得膏体；
5)将膏体置于真空搅拌机内，加入去离子水补足余量，真空搅拌均匀，包装即得。
实施例4
麦冬高异黄酮提取物4％、维生素B1 0.2％、浙麦冬甾体皂苷提取物0.3％、甘氨胆酸钠0.1％、卡波姆980 0.4％、透明质酸钠0.05％、甘油10％、乙二醇2％、茶油0.05％、吐温-80 0.05％、氮酮0.05％、尼泊金乙酯0.5％、尿囊素0.5％、壳聚糖4％，余量为去离子水。
制备方法：
1)A相的制备:将0.4重量份卡波姆980倒入60重量份去离子水中搅拌，待分散均匀后加入0.5重量份尿囊素和0.05重量份氮酮继续搅拌加热至80℃恒温15min；
2)B相的制备:在10重量份甘油中加入0.05重量份透明质酸钠搅拌，待透明质酸分散均匀后加入适量蒸馏水继续搅拌，待体系均匀后再加入0.05重量份茶油和2重量份乙二醇并加热搅拌至80℃恒温15min；
3)C相的制备:加入提取出来的4重量份浙麦冬高异黄酮提取物，0.3重量份浙麦冬甾体皂苷提取物、0.1重量份甘氨胆酸钠，0.2重量份维生素B1和适量水，并滴加少量乙醇，温热至55℃并用超声波溶解；
4)将B相组分加入到A相组分中，加入4重量份壳聚糖，于80℃搅拌15min后冷却。待冷却至55℃，加入溶解均一的C相组分，并使产品继续冷却。待产品冷却至45℃加入0.5重量份尼泊金乙酯，最后加入0.05重量份吐温-80，加去离子水，搅拌均匀即得膏体。
5)将膏体置于真空搅拌机内，加入去离子水补足余量，真空搅拌均匀，包装即得。
实施例5
麦冬高异黄酮提取物6％、维生素B1 0.2％、维生素C 0.2％、浙麦冬甾体皂苷提取物0.3％、甘氨胆酸钠0.1％、卡波姆980 0.2％、透明质酸钠0.01％、甘油18％、乙二醇1％、茶油0.05％、吐温-80 2％、氮酮2％、尼泊金乙酯0.1％，尿囊素0.05％，壳聚糖3％，余量为去离子水。
制备方法：
1)A相的制备:将0.2重量份卡波姆980倒入50重量份去离子水中搅拌，待分散均匀后加入0.05重量份尿囊素和2重量份氮酮继续搅拌加热至80℃恒温15min；
2)B相的制备:在18重量份甘油中加入0.01重量份透明质酸钠搅拌，待透明质酸分散均匀后加入适量蒸馏水继续搅拌，待体系均匀后再加入0.05重量份茶油和1重量份乙二醇并加热搅拌至80℃恒温15min；
3)C相的制备:加入提取出来的6重量份浙麦冬高异黄酮提取物，0.3重量份浙麦冬甾体皂苷提取物，0.1重量份甘氨胆酸钠、0.2重量份维生素B1和适量水，并滴加少量乙醇，温热至55℃并用超声波溶解；
4)将B相组分加入到A相组分中，加入3重量份壳聚糖，于80℃搅拌15min后冷却。待冷却至55℃，加入溶解均一的C相组分，并使产品继续冷却。待产品冷却至45℃加入0.1重量份尼泊金乙酯，最后加入2重量份吐温-80，加去离子水，搅拌均匀即得膏体；
5)将膏体置于真空搅拌机内，加入去离子水补足余量，真空搅拌均匀，包装即得。
实验例
实验例1：浙麦冬高异黄酮提取物中四种高异黄酮的含量分析
本实施例所采用分析浙麦冬高异黄酮提取物及产品中高异黄酮含量的HPLC条件为：Waters HPLC系统包括e2695四元泵分离单元，柱温箱和2998二极管阵列检测器；色谱柱Thermo C18色谱柱(4.6×250mm,5.0μm)；流动相由乙腈(A)和水(B)组成，梯度洗脱方案为：0～5min,35％A；5～20min,35～40％A；20～50min,40～70％A。柱温35℃；流速1mL/min，检测波长285nm。进样量根据样品浓度一般为5～20μL。4种浙麦冬高异黄酮标准品和样品的HPLC检测图谱见图1和2。
实验例2：浙麦冬高异黄酮提取物酪氨酸酶抑制活性研究
磷酸盐(pH 6.8，0.05mol/L)缓冲体配置：
酪氨酸酶溶液的配置：精密称取酪氨酸酶，用磷酸钠缓冲液配成5×10-6mol/L的酶液，-20℃冷冻保存，用前4℃解冻。
L-多巴(L-DOPA)溶液的配置：精密称取L-DOPA，加入磷酸盐缓冲液溶解，配得5.0×10-4mol/L的L-多巴溶液。
样品溶液的配置：将麦冬高异黄酮提取物，配制成不同浓度的高异黄酮复合液。
测定方法：酪氨酸酶催化底物L-DOPA生成的产物在475nm处特征吸收峰，利用抑制动力学方法，在3mL的磷酸盐(pH 6.8，0.05mol/L)缓冲体系中，固定酪氨酸酶的浓度2.08×10-7mol/L，将其与不同浓度的高异黄酮提取物在室温条件下孵化3h，使其充分反应后，加入底物L-DOPA(5.0×10-4mol/L)，每隔5s测定一次反应体系在475nm处的吸光度，通过关系式(1)，计算在含有不同浓度高异黄酮体系下酪氨酸酶的相对活性。曲酸作为阳性对照。式中R0、R分别为不加抑制剂和添加不同浓度抑制剂时反应体系的吸光度随时间的变化率。
相对活性(％)＝(R/R0)×100％ (1)
表1浙麦冬高异黄酮提取物酪氨酸酶活性抑制率
实验结果表明，浙麦冬高异黄酮提取物具有明显的酪氨酸酶活性抑制效果。说明本发明能有效减轻黄褐斑及痤疮引起的黑斑，具有祛斑美白、保湿的作用。
实验例3：浙麦冬多糖、水提取物的氨酸酶抑制活性研究
采用与实验例2相同的方法，对不同浓度浙麦冬多糖、麦冬水提物对酪氨酸酶抑制活性进行测定，结果见表2。
表2麦冬多糖、水提取物酪氨酸酶活性抑制率
实验结果表明，与浙麦冬高异黄酮提取物相比，浙麦冬多糖及水提物对酪氨酸酶活性抑制率较低，且各浓度对酪氨酸酶均无明显抑制作用，说明浙麦冬多糖、水提取物并非浙麦冬美白的有效成分。
实验例4：浙麦冬高异黄酮提取物与美白增强剂协同作用抑制酪氨酸酶活性研究
按本发明实施例中浙麦冬高异黄酮提取物与不同比例增强剂配制美白增强复合液。按照实施例1配制的浙麦冬高异黄酮提取物、美白增强剂复合液标记为复合液1，按照实施例2配制的浙麦冬高异黄酮提取物、美白增强剂复合液标记为复合液2，依此类推。采用与实验例1相同的方法测定复合液和浙麦冬高异黄酮提取物对酪氨酸酶的抑制活性，浙麦冬高异黄酮提取物浓度为3％(按生药量计算)，结果见表3。
表3各样品酪氨酸酶活性抑制率
实验例5：基础理化指标
参照国家标准QB/T1857-2013和QB/T2660-2004,对面膜的色泽、气味、涂展性、黏稠度、产品结构和稳定性进行测试,确定是否符合国家标准。本面膜试样的相关理化检测指标、感官及稳定性实验结果见表4。
表4理化检测指标、感官及稳定性实验结果
实验例6：皮肤刺激性测试
选择30名年龄为20～30岁的志愿者作为受试对象,在其左手背将面膜均匀涂抹,每天2次，6h后观察皮肤的反应。经过一周的实验观察，30名皮肤健康的在校大学生志愿者涂有面膜液的左手背皮肤比没有涂面膜液的右手背皮肤要柔软润泽一些，未见发红、起疹、起水泡等现象，也无发痒、过敏、疼痛等现象，得出该面膜无刺激性。
Claims (7)
1. 一种凝胶型浙麦冬美白面膜，其特征在于包括以下重量百分比的组分：浙麦冬高异黄酮提取物1-6%、吸收促进剂0.05-0.5%、络合剂1-7%、美白增强剂0.04-0.4%、卡波姆9800.2-0.8%、透明质酸钠0.01-0.05%、甘油10-20%、乙二醇1-5%、茶油0.05-0.1%、吐温-800.05-2%、氮酮0.05-2%、尼泊金乙酯0.1-0.5%、尿囊素0.05-0.5%、余量为去离子水；所述吸收促进剂由浙麦冬甾体皂苷提取物和胆酸盐组成；所述浙麦冬甾体皂苷与胆酸盐的质量比为3:1；浙麦冬甾体皂苷提取物按干燥品计算，含麦冬总皂苷以鲁斯可苷元计，不得少于5%，胆酸盐选用甘氨胆酸钠、甘氨鹅脱氧胆酸钠、牛磺胆酸钠、牛磺鹅脱氧胆酸钠、脱氧胆酸钠中的一种或两种以上；所述美白增强剂为维生素B1和维生素C中的一种或二种；
所述浙麦冬高异黄酮提取物和浙麦冬甾体皂苷提取物的制备方法为：
1）浙麦冬粉末加入10-20倍量的70%的乙醇，80℃加热回流提取1-2h，提取1-3次，滤液浓缩得到浙麦冬提取物浸膏，加去离子水稀释成浓度至每1ml相当于原有药材0.1-1g的水提取物；
2）取步骤1）的提取物，按提取物与树脂的固液比为1-10mL/g上样于D101大孔树脂，依次用蒸馏水、60%的乙醇、90%乙醇洗脱，收集90%乙醇洗脱液，减压浓缩，真空冷冻干燥得到提取物1，即浙麦冬高异黄酮提取物；其中，收集60%乙醇洗脱液，减压浓缩，真空冷冻干燥得到提取物2，即浙麦冬甾体皂苷提取物。
2. 如权利要求1所述的一种凝胶型浙麦冬美白面膜，其特征在于所述浙麦冬高异黄酮提取物6%、吸收促进剂0.4%、络合剂1%、美白增强剂 0.4%、卡波姆980 0.2%、透明质酸钠0.01%、甘油18%、乙二醇1%、茶油0.05%、吐温-80 2%、氮酮2%、尼泊金乙酯0.1%、尿囊素0.05%、余量为去离子水。
3.如权利要求1或2所述的一种凝胶型浙麦冬美白面膜，其特征在于所述浙麦冬高异黄酮提取物是指：浙麦冬高异黄酮提取物按干燥品计，所含甲基麦冬黄烷酮A，甲基麦冬黄烷酮B，6-醛基异麦冬黄烷酮A和6-醛基异麦冬黄烷酮B的总量不得少于10%。
4.如权利要求1或2所述的一种凝胶型浙麦冬美白面膜，其特征在于所述络合剂为EDTA-二钠或壳聚糖中的一种或二种，EDTA-二钠的重量百分比为2-3%，壳聚糖的重量百分比为1-4%。
5.权利要求1-4任一所述的一种凝胶型浙麦冬美白面膜的制备方法，其特征在于包括以下步骤：
1）A相的制备：将配方量的卡波姆980倒入适量去离子水中搅拌，待分散均匀后加入配方量的尿囊素和氮酮继续搅拌加热至 80℃恒温15min；
2）B相的制备：在配方量的甘油中加入配方量的透明质酸钠搅拌，待透明质酸分散均匀后加入适量蒸馏水继续搅拌，待体系均匀后再加入配方量的茶油和乙二醇并加热搅拌至80℃恒温 15min；
3）C相的制备：将配方量的浙麦冬高异黄酮提取物、促进吸收剂、美白增强剂加入适量的去离子水中，并滴加少量乙醇，温热至 55℃并用超声波溶解；
4）将B相组分加入到 A相组分中，加入配量的络合剂，于80℃ 搅拌15min后冷却至 55℃，加入溶解均一的C相组分，并使产品继续冷却至45℃ ，然后依次加入配方量的尼泊金乙酯和吐温-80，加去离子水至 100mL，搅拌均匀即得膏体；
5）将膏体置于真空搅拌机内，加入去离子水补足余量，真空搅拌均匀，包装即得凝胶型浙麦冬美白面膜。
6.如权利要求5所述的一种凝胶型浙麦冬美白面膜的制备方法，其特征在于步骤1）中适量去离子水占去去离子水总量的70-80%，步骤4）中络合剂为EDTA-二钠和壳聚糖两者中的一种或两种。
7.权利要求1-6任一所述的一种凝胶型浙麦冬美白面膜在制备减轻黄褐斑及痤疮引起的黑斑产品上应用。
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