CN114213676A - 一种天然深共晶紫苏叶提取物水凝胶的制备方法 - Google Patents
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Abstract
本发明公开了一种天然深共晶紫苏叶提取物水凝胶的制备方法。本发明的制备方法包括：以紫苏叶提取物为原料，加入天然深共晶溶液、多巴胺、甲基丙烯酸羟乙酯和2‑(二甲基胺)‑甲基丙烯酸乙酯充分混匀，30～40℃下交联反应制得基于天然深共晶紫苏叶提取物的伤口敷料水凝胶；所述天然深共晶溶液由摩尔比为1:2的甘露糖和氯化胆碱组成。本发明利用甘露糖和氯化胆碱组成的天然深共晶溶液对紫苏叶活性成分的高提取效率，以及该天然深共晶溶液体系中的氢键与单体形成的三维复合网络，能够增进水凝胶网络结构的稳固性，从而制备得到一种具有良好的物理性能和抑菌性能的水凝胶，可用作医用伤口敷料，具有良好的应用前景。
Description
技术领域
本发明涉及一种天然深共晶紫苏叶提取物水凝胶的制备方法，属于医用材料技术领域。
背景技术
水凝胶是一类具有三维网络结构的亲水性高分子材料，能够吸收高达自身重量70％的水分溶胀又不溶解，具有良好的生物相容性、与细胞外基质的高度相似性，可支持细胞的增殖和迁移，高度模拟软组织。水凝胶近几年在生物医用材料领域备受关注，主要应用存在且不限于创伤敷料、人体软组织填充、角膜接触镜、载药基质、3D器官打印等，且适用范围也不断扩大至食品工业、化学阀门、机械制造等领域。就伤口敷料来说，传统的伤口敷料与组织不贴和、透气性差、给药不均，水凝胶作为伤口敷料则就具有高渗透性且贴合损伤组织，模拟其愈合的湿环境，降低了组织感染的风险，提高了伤口愈合速度。但就目前水凝胶研究而言，水凝胶作为伤口敷料，稳定附着力和结构支撑强度、耐损性仍有待改进，添加的功能性物质如何与水凝胶基质共存且发挥良好效用的问题也有待解决。
天然深共晶溶液由天然的生物代谢产物(如葡萄糖、乳酸、柠檬酸、甘油等)组成，其优良的提取性能和稳定性来源于分子间氢键的作用力。天然深共晶溶液与生物组织有较好的相容性，相较于传统溶液能更高更有效地提取天然原料中的有效生物活性成分。且其性状具有可塑造性和可设计性，可通过成分之间的配比、调节溶液pH等方法来调节其流变性能。紫苏叶在中国具有悠久的栽种历史，是药食同源的植物之一。紫苏叶的提取物具有较好的抗菌、消炎等功效。
目前未见有天然深共晶溶液的天然活性提取物用于制备三维网状水凝胶的相关报道。
发明内容
本发明解决的技术问题是：如何利用天然深共晶紫苏叶提取物制备具有良好的生物相容性、物理性能和杀菌性能的水凝胶的问题。
为了解决上述技术问题，本发明提供了一种天然深共晶紫苏叶提取物水凝胶的制备方法，包括：以紫苏叶提取物为原料，加入天然深共晶溶液、多巴胺、甲基丙烯酸羟乙酯和2-(二甲基胺)-甲基丙烯酸乙酯充分混匀，30～40℃下交联反应制得基于天然深共晶紫苏叶提取物的伤口敷料水凝胶；所述天然深共晶溶液由摩尔比为1:2的甘露糖和氯化胆碱组成。
优选地，所述的紫苏叶提取物为紫苏叶经水提后冻干得到的紫苏叶水提物粉末。
优选地，所述紫苏叶提取物与天然深共晶溶液的质量比为1:2～6；所述紫苏叶提取物与多巴胺、甲基丙烯酸羟乙酯和2-(二甲基胺)-甲基丙烯酸乙酯的质量比为2：0.3～3：0.1～0.3：0.1～0.4。
优选地，所述交联反应的时间为20～30h。
本发明还提供了上述的天然深共晶紫苏叶提取物水凝胶的制备方法制备所得的天然深共晶紫苏叶提取物水凝胶，所述的天然深共晶紫苏叶提取物水凝胶对大肠杆菌的抑菌浓度≥10mg/mL。
本发明的原理：
甘露糖参与人体新陈代谢，作为天然深共晶溶液的组成之一，具有消炎作用。氯化胆碱是天然深共晶溶液中常用的季铵盐，也作为一种营养补充剂，具有杀菌作用。且该天然深共晶溶液体系中的氢键与单体形成三维复合网络，增进水凝胶网络结构的稳固性。甲基丙烯酸羟乙酯(HEMA)和2-(二甲基胺)-甲基丙烯酸乙酯(DMAEMA)用于形成水凝胶的自聚合网络结构。多巴胺作水凝胶粘合剂。
本发明与现有技术相比，具有如下有益效果：
1.本发明的一种天然深共晶紫苏叶提取物水凝胶的制备方法，利用甘露糖和氯化胆碱组成的天然深共晶溶液对紫苏叶活性成分的高提取效率，以及该天然深共晶溶液体系中的氢键与单体形成的三维复合网络，能够增进水凝胶网络结构的稳固性，从而制备得到一种具有良好的物理性能和抑菌性能的水凝胶；
2.本发明选材天然且具有高度生物相容性，对人体安全无毒，后续处理方便简易，因此本发明制备的天然深共晶紫苏叶提取物水凝胶可用作医用伤口敷料，具有良好的应用前景。
附图说明
图1为实施例2制备的天然深共晶紫苏叶提取物水凝胶的红外谱图；
图2为实施例2制备的天然深共晶紫苏叶提取物水凝胶的物理性能测试结果示意图；
图3为实施例2制备的天然深共晶紫苏叶提取物水凝胶的抑菌性能测试结果示意图。
具体实施方式
为使本发明更明显易懂，兹以优选实施例，并配合附图作详细说明如下。
实施例1紫苏叶提取物制备
将新鲜紫苏叶洗净后与蒸馏水混合，质量比紫苏叶：蒸馏水为1:20。混合物进行闪式提取后过滤、5000r/min离心15min，取上层清液冻干成紫苏叶水提物粉末。
实施例2天然深共晶紫苏叶提取物水凝胶的制备
将甘露糖-氯化胆碱(摩尔比1:2)5g混合，加热至85℃，匀速搅拌1h，得到澄清透明粘稠液体。加入实施例1中紫苏叶提取物2g，2.06g多巴胺，0.142g甲基丙烯酸羟乙酯(HEMA)和0.171g 2-(二甲基胺)-甲基丙烯酸乙酯(DMAEMA)充分混匀，35℃交联24h，得到水凝胶A。
实施例3天然深共晶紫苏叶提取物水凝胶的制备
将甘露糖-氯化胆碱(摩尔比1:2)10g混合，加热至85℃，匀速搅拌1h，得到澄清透明粘稠液体。加入实施例1中紫苏叶提取物2g，1.412g多巴胺，0.284g甲基丙烯酸羟乙酯(HEMA)和0.342g 2-(二甲基胺)-甲基丙烯酸乙酯(DMAEMA)充分混匀，35℃交联24h，得到水凝胶B。
实施例4天然深共晶紫苏叶提取物水凝胶的制备
将甘露糖-氯化胆碱(摩尔比1:2)10g混合，加热至85℃，匀速搅拌1h，得到澄清透明粘稠液体。加入实施例1中紫苏叶提取物2g，2.824g多巴胺，0.284g甲基丙烯酸羟乙酯(HEMA)和0.342g 2-(二甲基胺)-甲基丙烯酸乙酯(DMAEMA)充分混匀，35℃交联24h，得到水凝胶C。
实施例5天然深共晶紫苏叶提取物水凝胶的制备
将甘露糖-氯化胆碱(摩尔比1:2)10g混合，加热至85℃，匀速搅拌1h，得到澄清透明粘稠液体。加入实施例1中紫苏叶提取物2g，0.355多巴胺，0.142g甲基丙烯酸羟乙酯(HEMA)和0.342g 2-(二甲基胺)-甲基丙烯酸乙酯(DMAEMA)充分混匀，35℃交联24h，得到水凝胶D。
实施例6天然深共晶紫苏叶提取物水凝胶的红外表征
使用VERTEX研究级傅立叶变换红外光谱仪(德国，Bruker)对实施例2制备的水凝胶A进行红外表征。结果显示如图1所示。由图可得，在2887cm-1存在HEMA\DMAEMA单元拉伸振动；1730cm-1存在HEMA\DMAEMA的–C＝O伸缩振动；1300-1000cm-1存在C-O-C不对称伸缩振动。3500-3000cm-1显示出氯化胆碱和甘露糖之间分子间氢键的强峰；1636cm-1显示出多巴胺中苯环的C-H弯曲振动。证明已合成水凝胶。
实施例7天然深共晶紫苏叶提取物水凝胶的多功能物理性能测定
取实施例2制备的水凝胶A进行斜置、剪切、拉伸的物理性能测试。结果如图2所示。由图2可知，30天后凝胶的颜色加深，仍然具有较强的凝胶物理性能。该凝胶具有一定的流动性，并具有可切断性和较优的拉伸性。
实施例8天然深共晶紫苏叶提取物水凝胶对大肠杆菌的抑菌性能测定
通过共培养和滤纸片法进行水凝胶对于大肠杆菌的抑菌测定。取实施例2制备的水凝胶A配制成浓度为30mg/mL、25mg/mL、20mg/mL、15mg/mL、10mg/mL、5mg/mL、2.5mg/mL、1.25mg/mL、0.625mg/mL、0.3125mg/mL的溶液，溶剂为超纯水。
与大肠杆菌的共培养实验：大肠杆菌于37℃、200r/min振荡培养至106～107CFU，分别取上述浓度凝胶溶液0.9mL与0.1mL菌液混合，并以0.9mL生理盐水与0.1mL菌液混合作为对照。实验组与对照组于37℃下培养10h后，取菌-凝胶混合液0.1mL涂布于琼脂固体培养基上，37℃下培养12h。
滤纸片法：6mm滤纸片负载0.02g水凝胶后备用。大肠杆菌于37℃、200r/min振荡培养至106～107CFU，取0.1mL菌液涂布于琼脂固体培养基上，后加入滤纸片于培养基上，37℃下培养24h，平行进行三次实验。
性能测试结果如图3所示，由图3可知，共培养法中，水凝胶A的浓度小于10mg/mL的各试验组以及对照组中大肠杆菌于培养皿中生长，滤纸片法测得水凝胶A的平均抑菌圈为12.28mm，证明本发明制备的基于天然深共晶紫苏叶提取物水凝胶对于大肠杆菌有抑制作用，其抑菌浓度≥10mg/mL。
以上所述，仅为本发明的较佳实施例，并非对本发明任何形式上和实质上的限制，应当指出，对于本技术领域的普通技术人员，在不脱离本发明的前提下，还将可以做出若干改进和补充，这些改进和补充也应视为本发明的保护范围。
Claims (5)
1.一种天然深共晶紫苏叶提取物水凝胶的制备方法，其特征在于，包括：以紫苏叶提取物为原料，加入天然深共晶溶液、多巴胺、甲基丙烯酸羟乙酯和2-(二甲基胺)-甲基丙烯酸乙酯充分混匀，30～40℃下交联反应制得天然深共晶紫苏叶提取物水凝胶；所述天然深共晶溶液由摩尔比为1:2的甘露糖和氯化胆碱组成。
2.如权利要求1所述的天然深共晶紫苏叶提取物水凝胶的制备方法，其特征在于，所述的紫苏叶提取物为紫苏叶经水提后冻干得到的紫苏叶水提物粉末。
3.如权利要求2所述的天然深共晶紫苏叶提取物水凝胶的制备方法，其特征在于，所述紫苏叶提取物与天然深共晶溶液的质量比为1:2～6；所述紫苏叶提取物与多巴胺、甲基丙烯酸羟乙酯和2-(二甲基胺)-甲基丙烯酸乙酯的质量比为2：0.3～3：0.1～0.3：0.1～0.4。
4.如权利要求1所述的天然深共晶紫苏叶提取物水凝胶的制备方法，其特征在于，所述交联反应的时间为20～30h。
5.权利要求1～4中任意一项所述的天然深共晶紫苏叶提取物水凝胶的制备方法制备所得的天然深共晶紫苏叶提取物水凝胶，其特征在于，所述的天然深共晶紫苏叶提取物水凝胶对大肠杆菌的抑菌浓度≥10mg/mL。
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