CN113797118A - 一种包载芋螺毒素的壳聚糖纳米粒乳液及其制备方法 - Google Patents
一种包载芋螺毒素的壳聚糖纳米粒乳液及其制备方法 Download PDFInfo
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Abstract
本发明公开一种包载芋螺毒素的壳聚糖纳米粒乳液及其制备方法，属于高分子材料技术领域。乳液包括以下重量份的原料制备而成：芋螺毒素1份、壳聚糖2‑4份、三聚磷酸钠0.1‑0.5份、油脂性成分100‑400份、乳化剂40‑160份、辅助乳化剂80‑200份、保湿剂80‑160份、增稠剂10‑30份。本发明使用壳聚糖包覆芋螺毒素制备纳米颗粒，并制备为乳液，促进芋螺毒素不断渗透进入皮肤深层发挥祛皱作用；同时添加有辅助乳化剂、保湿剂等成分，有助于皮肤角质层的水合作用，增强芋螺毒素纳米颗粒的经皮渗透；各原料共同作用，相互配合，共同提升了芋螺毒素的透皮渗透率，促进芋螺毒素进一步的实际应用和推广，经济价值显著。
Description
技术领域
本发明属于高分子材料技术领域，具体涉及一种包载芋螺毒素的壳聚糖纳米粒乳液及其制备方法。
背景技术
衰老是一种自然现象，随着年龄的增长，人类的皮肤也会逐渐老化，最明显的表现就是面部皱纹的增多、加深，人们一直希望通过各种方法减缓皮肤衰老，使容颜永驻，永葆青春。祛除面部皱纹无疑是保持青春容颜的最直接有效的方式，因此祛皱产品引起了美容护肤专业人士的高度关注。
在祛皱产品中，肽类产品占据着重要的地位。芋螺毒素由12-40个氨基酸组成，富含二硫键，是由芋螺毒液管和毒囊内壁的毒腺所分泌，其可作用于肌肉型钠离子阀门通道Nav 1.4，阻断神经肌肉的电流传导让横纹肌放松下来，进而瞬间淡化皱纹。芋螺毒素放松横纹肌但不会导致肌肉僵硬瘫痪，因此可使皮肤达到自然回春的效果。
然而芋螺毒素透皮渗透能力差，皮肤涂抹难以使芋螺毒素渗透进入皮肤深层，限制了其祛皱作用的发挥。增加活性物质透皮转运的方法包括物理方法、化学方法及药剂学方法。物理方法主要通过粒子导入、超声波导入等方式实现活性物质的经皮渗透，化学方法主要通过吸收促进剂来增加活性物质的经皮渗透，然而物理方法往往需要借助特殊导入设备，不利于消费者日常自行使用。化学方法使用的吸收促进剂对皮肤具有不同程度的刺激性。
因此，药剂学方法越来越受到人的关注。药剂学方法通常借助于脂质体、纳米粒、微乳等纳米载体将活性物质输送进入皮肤深层，具有生物相容性好、无毒、无刺激等特点。
发明内容
本发明针对现有技术所存在的缺陷，提供一种包载芋螺毒素的壳聚糖纳米粒乳液，将芋螺毒素包埋与壳聚糖纳米颗粒中，进一步制备芋螺毒素壳聚糖纳米粒乳液，以大幅提升芋螺毒素渗透性，且安全无毒无刺激。
为实现上述技术目的，本发明所采用的技术方案为：
一种包载芋螺毒素的壳聚糖纳米粒乳液，包括以下重量份的原料制备而成：芋螺毒素1份、壳聚糖2-4份、三聚磷酸钠0.1-0.5份、油脂性成分100-400份、乳化剂40-160份、辅助乳化剂80-200份、保湿剂80-160份、增稠剂10-30份。
进一步的，所述油脂性成分为液体石蜡、肉豆蔻酸异丙酯、维生素E、棕榈酸异丙酯中的一种或几种。
进一步的，所述乳化剂为吐温80和/或15-羟基硬脂酸聚乙二醇。
进一步的，所述辅助乳化剂为硬脂酸、大豆磷脂、单硬脂酸甘油酯、丙二醇中的一种或几种。
进一步的，所述保湿剂为透明质酸、卡波姆934、卡波姆940、聚乙二醇1000、甘油中的一种或几种。
进一步的，所述增稠剂为羧甲基纤维素钠和/或甲基纤维素。
一种包载芋螺毒素的壳聚糖纳米粒乳液的制备方法，包括以下步骤：
(1)将芋螺毒素和壳聚糖溶解于乙酸溶液中，磁力搅拌下缓慢滴加到三聚磷酸钠的水溶液中，滴加完毕调节pH至5.5，继续搅拌得芋螺毒素壳聚糖纳米粒混悬液；
(2)再向混悬液中加入乳化剂、保湿剂和增稠剂，溶解混匀，得到水相混合物，70℃保温备用；
(3)取辅助乳化剂和油脂性成分于容器中，水浴加热至70℃使熔融，得到油相混合物；
(4)将油相混合物缓慢加入水相混合物，边加边搅拌，冷却至室温即得包载芋螺毒素的壳聚糖纳米粒乳液。
进一步的，步骤(1)芋螺毒素和壳聚糖固体混合物和乙酸溶液的固液比为3mg:1ml，乙酸溶液的质量浓度为1％。
进一步的，步骤(1)三聚磷酸钠的水溶液的浓度为0.1mg/ml。
壳聚糖是一种由甲壳素脱乙酰基而得到的天然阳离子多糖，对人体无毒、无刺激，因而具有良好的生物可降解性，也经常被用于伤口包扎辅料中。
有益效果
(1)本发明使用壳聚糖包覆芋螺毒素制备纳米颗粒，壳聚糖属于一种聚阳离子多糖，芋螺毒素壳聚糖纳米颗粒表面带正电荷，而皮肤表面带负电荷，芋螺毒素壳聚糖纳米粒与皮肤亲和性强；壳聚糖纳米颗粒与皮肤接触后，通过皮肤毛孔在毛囊处聚集形成贮库，促进芋螺毒素不断渗透进入皮肤深层发挥祛皱作用；
(3)本发明将芋螺毒素的混悬液制备为乳液，乳液本身具有一定的粘稠度，铺展性好，方便涂抹，与皮肤亲和力强，有助于芋螺毒素纳米颗粒与皮肤有效接触；本发明还添加有辅助乳化剂、保湿剂等成分，有助于皮肤角质层的水合作用，增强芋螺毒素纳米颗粒的经皮渗透；各原料共同作用，相互配合，共同提升了芋螺毒素的透皮渗透率，促进芋螺毒素进一步的实际应用和推广，经济价值显著。
附图说明
图1为本发明实施例1乳液中芋螺毒素壳聚糖纳米颗粒扫描电镜照片；
图2为本发明实施例1乳液中芋螺毒素壳聚糖纳米颗粒体外释曲线；
图3为本发明实施例1、对比例2、对比例3乳液除皱效果试验图。
具体实施方式
下面结合具体实施例对本发明的技术方案做进一步说明，但不限于此。
实施例1
取50mg芋螺毒素，100mg壳聚糖溶于50ml 1％的乙酸溶液，磁力搅拌下缓慢滴入0.1mg/ml三聚磷酸钠溶液50ml，滴加完毕调节pH至5.5，继续搅拌片刻即得芋螺毒素壳聚糖纳米粒混悬液，加入吐温80 2g、透明质酸2g、甘油2g、羧甲基纤维素钠0.5g溶解，混匀，得到水相，70℃保温备用；另取硬脂酸2g、单硬脂酸甘油脂2g、肉豆蔻酸异丙酯4.5g、维生素E0.5g、放于烧杯中，水浴加热至70℃使其熔融，得到油相；将制得的油相缓缓加入水相，边加边搅拌，放冷至室温即得包载芋螺毒素的壳聚糖纳米粒乳液。
实施例2
取50mg芋螺毒素，200mg壳聚糖溶于50ml 1％的乙酸溶液，磁力搅拌下缓慢滴入0.5mg/ml三聚磷酸钠溶液50ml，滴加完毕调节pH至5.5，继续搅拌片刻即得芋螺毒素壳聚糖纳米粒混悬液。加入15-羟基硬脂酸聚乙二醇8g、聚乙二醇1000 2g、甘油6g、甲基纤维素1.5g溶解，混匀，得到水相，70℃保温备用；另取大豆磷脂4g、单硬脂酸甘油脂6g、棕榈酸异丙酯19g、液体石蜡1g、放于烧杯中，水浴加热至70℃使其熔融，得到油相；将制得的油相缓缓加入水相，边加边搅拌，放冷至室温即得包载芋螺毒素的壳聚糖纳米粒乳液。
实施例3
取50mg芋螺毒素，150mg壳聚糖溶于50ml 1％的乙酸溶液，磁力搅拌下缓慢滴入0.3mg/ml三聚磷酸钠溶液50ml，滴加完毕调节pH至5.5，继续搅拌片刻即得芋螺毒素壳聚糖纳米粒混悬液。加入15-羟基硬脂酸聚乙二醇8g、卡波姆940 1g、甘油4g、甲基纤维素1.0g溶解，混匀，得到水相，70℃保温备用；另取大豆磷脂2g、硬脂酸1g、丙二醇5g、棕榈酸异丙酯10g、肉豆蔻酸异丙酯5g、放于烧杯中，水浴加热至70℃使其熔融，得到油相；将制得的油相缓缓加入水相，边加边搅拌，放冷至室温即得包载芋螺毒素的壳聚糖纳米粒乳液。
对比例1
取50mg芋螺毒素溶解于150ml纯水即得芋螺毒素溶液。
对比例2
取50mg芋螺毒素，加入吐温80 2g、透明质酸2g、甘油2g、羧甲基纤维素钠0.5g溶解，混匀，得到水相，70℃保温备用；另取硬脂酸2g、单硬脂酸甘油脂2g、肉豆蔻酸异丙酯4.5g、维生素E0.5g、放于烧杯中，水浴加热至70℃使其熔融，得到油相；将制得的油相缓缓加入水相，边加边搅拌，放冷至室温即得芋螺毒素壳聚糖纳米颗粒乳液。
对比例3
取50mg芋螺毒素，100mg壳聚糖溶于50ml 1％的乙酸溶液，磁力搅拌下缓慢滴入0.1mg/ml三聚磷酸钠溶液50ml，滴加完毕调节pH至5.5，继续搅拌片刻加水定容至150ml即得芋螺毒素壳聚糖纳米粒混悬液。
性能测试
(1)形貌测试
以实施例1所得乳液为例，采用扫描电镜观察乳液中芋螺毒素壳聚糖纳米颗粒外观形态如图1所示，由扫描电镜照片可见，螺毒素壳聚糖纳米颗粒是一种实体小球。经马尔文激光粒度仪测定实施例1芋螺毒素壳聚糖纳米颗粒粒径为93.2±7.6nm，颗粒表面带正电荷，zeta电位为40.5±5mv。
(2)体外释放度测定
对实施例1芋螺毒素壳聚糖纳米颗粒进行体外释放度测定，方法如下：实施例1包载芋螺毒素壳聚糖纳米颗乳液0.2ml，加1％乙酸溶液稀释并定容至10ml，超声处理10min，测定芋螺毒素总含量(C总)。另取芋螺毒素壳聚糖纳米颗乳液0.2ml，分别加入pH 5.5的醋酸盐缓冲液5mL(37℃恒温)，在0.5、1、2、3、5、8、12、18、24h取样，离心(相对离心力350g)5min，取上清液测定游离芋螺毒素含量(C游)。
芋螺毒素体外释放曲线如图2所示，结果显示，芋螺毒素聚糖纳米颗粒释放平稳缓慢，24h释放完全。
(3)透皮性能评价
采用改良的Franz扩散池，透皮扩散面直径15mm，透皮扩散面积1.77cm2，接收室体积15ml。取小鼠腹部皮肤固定在供给池和接收池间，角质层面向供给池。将生理盐水接收液注入接收池，水浴温度为(32±0.5)℃，于300r·min-1恒速搅拌。分别对实施例1及对比例1、对比例2、对比例3进行透皮实验(每份样品平行3次实验)。实验终止后，取下鼠皮，用蒸馏水洗净皮肤，将皮肤剪碎加入5ml1％醋酸溶液浸泡6h，高速离心，将上清液冷冻干燥，用少量流动相溶解，测定芋螺毒素含量，计算8小时不同剂型皮肤滞留量。测试结果如表1所示：
表1皮肤滞留量测试结果
| 皮肤滞留量(μg/cm<sup>2</sup>) | P value | |
| 实施例1 | 155.36 | <0.01 |
| 对比例1 | 22.34 | <0.01 |
| 对比例2 | 35.33 | <0.01 |
| 对比例3 | 78.79 | <0.01 |
*P<0.01代表与实施例1相比具有显著性差异。
由结果可见，皮肤滞留量大小顺序为实施例1＞对比例3>对比例2>对比例1,说明本发明实施例芋螺毒素纳米颗粒乳液渗透性最强，而芋螺毒素壳聚糖纳米颗粒渗透性强于芋螺毒素乳液及芋螺毒素溶液，芋螺毒素溶液的皮肤渗透性最差。统计学检验表明，具体对比例三种剂型与实施例1的差别均具有统计学意义，说明本发明壳聚糖纳米颗粒乳液显著提高了芋螺毒素的透皮渗透。
(4)除皱效果评价
健康志愿者18例，眼角细纹明显，试验2周内没有使用任何对结果有影响的化妆品、药物和保健品。将志愿者随机分为3组，每组6人，分别于左眼眼角涂抹实施例1、对比例2、对比例3样品，分别于使用前0h及使用后2、4、6、8、12、24h采用VISIA皮肤检测仪(美国Canfield公司)对受试者进行图像采集，测定使用前后眼部皱纹的分值，评价抗皱效果，分值越低说明除皱效果越明显。结果如下图3所示，本发明实施例所得芋螺毒素乳液分值较低，呈现了良好的除皱效果。
需要说明的是，上述实施例仅仅是实现本发明的优选方式的部分实施例，而非全部实施例。显然，基于本发明的上述实施例，本领域普通技术人员在没有做出创造性劳动的前提下所获得的其他所有实施例，都应当属于本发明保护的范围。
Claims (9)
1.一种包载芋螺毒素的壳聚糖纳米粒乳液，其特征在于，包括以下重量份的原料制备而成：芋螺毒素1份、壳聚糖2-4份、三聚磷酸钠0.1-0.5份、油脂性成分100-400份、乳化剂40-160份、辅助乳化剂80-200份、保湿剂80-160份、增稠剂10-30份。
2.根据权利要求1所述包载芋螺毒素的壳聚糖纳米粒乳液，其特征在于，所述油脂性成分为液体石蜡、肉豆蔻酸异丙酯、维生素E、棕榈酸异丙酯中的一种或几种。
3.根据权利要求1所述包载芋螺毒素的壳聚糖纳米粒乳液，其特征在于，所述乳化剂为吐温80和/或15-羟基硬脂酸聚乙二醇。
4.根据权利要求1所述包载芋螺毒素的壳聚糖纳米粒乳液，其特征在于，所述辅助乳化剂为硬脂酸、大豆磷脂、单硬脂酸甘油酯、丙二醇中的一种或几种。
5.根据权利要求1所述包载芋螺毒素的壳聚糖纳米粒乳液，其特征在于，所述保湿剂为透明质酸、卡波姆934、卡波姆940、聚乙二醇1000、甘油中的一种或几种。
6.根据权利要求1所述包载芋螺毒素的壳聚糖纳米粒乳液，其特征在于，所述增稠剂为羧甲基纤维素钠和/或甲基纤维素。
7.一种权利要求1-6任意一项所述包载芋螺毒素的壳聚糖纳米粒乳液的制备方法，其特征在于，包括以下步骤：
(1)将芋螺毒素和壳聚糖溶解于乙酸溶液中，磁力搅拌下缓慢滴加到三聚磷酸钠的水溶液中，滴加完毕调节pH至5.5，继续搅拌得芋螺毒素壳聚糖纳米粒混悬液；
(2)再向混悬液中加入乳化剂、保湿剂和增稠剂，溶解混匀，得到水相混合物，70℃保温备用；
(3)取辅助乳化剂和油脂性成分于容器中，水浴加热至70℃使熔融，得到油相混合物；
(4)将油相混合物缓慢加入水相混合物，边加边搅拌，冷却至室温即得包载芋螺毒素的壳聚糖纳米粒乳液。
8.根据权利要求7所述包载芋螺毒素的壳聚糖纳米粒乳液的制备方法，其特征在于，步骤(1)芋螺毒素和壳聚糖固体混合物和乙酸溶液的固液比为3mg:1ml，乙酸溶液的质量浓度为1％。
9.根据权利要求7所述包载芋螺毒素的壳聚糖纳米粒乳液的制备方法，其特征在于，步骤(1)三聚磷酸钠的水溶液的浓度为0.1mg/ml。
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| CN115353555A (zh) * | 2022-08-22 | 2022-11-18 | 山东大学 | 季铵化壳寡糖-μ-芋螺毒素结合物及其制备方法与应用 |
| CN116217693A (zh) * | 2023-01-06 | 2023-06-06 | 广东扶生肽生物科技有限公司 | 一种芋螺毒素外泌体及其应用 |
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| CN115300611B (zh) * | 2022-08-19 | 2024-05-28 | 山东大学 | 透明质酸-芋螺多肽结合物及其在制备皮肤产品中的应用 |
| CN115353555A (zh) * | 2022-08-22 | 2022-11-18 | 山东大学 | 季铵化壳寡糖-μ-芋螺毒素结合物及其制备方法与应用 |
| CN115353555B (zh) * | 2022-08-22 | 2024-04-30 | 山东大学 | 季铵化壳寡糖-μ-芋螺毒素结合物及其制备方法与应用 |
| CN116217693A (zh) * | 2023-01-06 | 2023-06-06 | 广东扶生肽生物科技有限公司 | 一种芋螺毒素外泌体及其应用 |
| CN116217693B (zh) * | 2023-01-06 | 2023-11-03 | 广东扶生肽生物科技有限公司 | 一种芋螺毒素外泌体及其应用 |
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