CN114052027A - 壳聚糖多维改性抗菌剂及其制备方法 - Google Patents
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A—HUMAN NECESSITIES
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Abstract
本发明提供一种壳聚糖抗菌剂，该抗菌剂主要由壳聚糖、鼠李糖脂、纳米二氧化钛、交联剂、稳定剂等原料多维改性复合而成，具有强抗菌性能。所述壳聚糖为低分子壳聚糖、脱乙酰度≥80%，具有广谱抗菌性、生物降解性、成膜性等特点，与纳米二氧化钛通过交联剂复合可显著提高抗菌剂在可见光下的抗菌性能，然后与生物表面活性剂鼠李糖脂形成复合抗菌剂，该复合抗菌剂具有抗菌力强、抗菌谱广、安全温和、性能稳定、成本低廉、绿色环保的特点。
Description
技术领域
本发明属于生物材料技术领域，具体涉及一种壳聚糖多维改性抗菌剂及其制备方法。
背景技术
壳聚糖是自然界中唯一的一种碱性多糖，是由甲壳素部分或全部脱除乙酰基后得到的。壳聚糖是自然界中含量仅次于纤维素的第二大丰富的生物多糖，是一种可再生的资源。壳聚糖不仅具有很高的黏度、良好的吸水性、保湿性、成膜性、增稠性、凝胶性等，还具有优异的生物相容性、无毒、抗菌性和生物降解性等特点，已成为一种新型的生理功能材料而广泛应用于医药、环保、纺织、食品及化妆品行业等领域。
国内外研究者对壳聚糖进行了改性，希望在抗菌方面提高其抗菌性能。目前，对于壳聚糖改性后用在抗菌方面的方法有：壳聚糖酯化改性、壳聚糖酰基化改性、壳聚糖季铵化改性、壳聚糖聚合物接枝改性和壳聚糖席夫碱改性等方法。但主要的应用领域集中于生物制剂或将壳聚糖与重金属复合形成的复合抗菌剂，需要用到大量有机溶剂，对于环境污染大，制备过程复杂，制备成本高，均不适用于工业化生产。
本发明采用壳聚糖交联多维改性方法，将无毒无害、无刺激性气味的无机纳米材料纳米二氧化钛与壳聚糖通过交联剂复合，再加入具有高降解性能的生物表面活性剂鼠李糖脂，形成性能稳定、抗菌力强、具有多维空间结构的壳聚糖/二氧化钛/鼠李糖脂改性复合抗菌剂，并且制备成本低廉、在制备和使用过程更加环保。
发明内容
针对上述现有技术中存在的问题，本发明提供一种壳聚糖多维改性抗菌剂及其制备方法，制备成本低廉，制备过程环保。本发明提供如下技术方案：
一种壳聚糖多维改性抗菌剂，其特征在于，该抗菌剂包括壳聚糖、鼠李糖脂、纳米二氧化钛、交联剂、稳定剂。
根据权利要求1所述一种壳聚糖多维改性抗菌剂，其特征在于，所述壳聚糖为低分子壳聚糖，分子量在10000以内，脱乙酰度≥80%。
根据权利要求1所述一种壳聚糖多维改性抗菌剂，其特征在于，所述鼠李糖脂是一种棕色膏状或白色粉末状的生物表面活性剂，纯度90%以上，具有抗菌功效。
根据权利要求1所述一种壳聚糖多维改性抗菌剂，其特征在于，所述纳米二氧化钛属于锐钛型纳米二氧化钛，是一种白色疏松粉末，直径在100纳米以下，具有抗菌、自洁净、老化性能。
根据权利要求1所述一种壳聚糖多维改性抗菌剂，其特征在于，所述交联剂为三聚磷酸钠、戊二醛、氯化钙等。
根据权利要求1所述一种壳聚糖多维改性抗菌剂，其特征在于，所述稳定剂包括有明胶、甘油、聚山梨酯类、山梨醇、聚乙二醇等。
根据权利要求1-6任一项所述壳聚糖多维改性抗菌剂的制备方法，依次包括下述步骤：（1）以纳米二氧化钛作为改性剂加入到壳聚糖醋酸溶液中，然后缓慢滴加适量的交联剂，保持30-40滴/min，同时在磁力搅拌器转速550r/min条件下反应2h，形成壳聚糖/二氧化钛乳状液；（2）将乳状液与鼠李糖脂溶液以体积比1:1进行混合，再加入0.5%的稳定剂，调整PH值为4～6，在磁力搅拌器转速650r/min、25℃条件下反应24h，合成壳聚糖/二氧化钛/鼠李糖脂三聚络合物。
根据权利要求7所述的壳聚糖多维改性抗菌剂制备方法，其特征在于，壳聚糖和鼠李糖脂溶液浓度范围在0.5～3.5mg/ml之间。
根据权利要求7所述的壳聚糖多维改性抗菌剂制备方法，其特征在于，壳聚糖混合液中需保持壳聚糖与交联剂的质量比在2:1～6:1范围内。
该壳聚糖多维改性复合抗菌剂具有抗菌力强、抗菌谱广、安全温和、性能稳定、成本低廉、绿色环保的特点，可应用于人类身体部位的清洁、抑菌。
具体实施方式
实施例1：一种壳聚糖多维改性抗菌剂，其成分为壳聚糖、鼠李糖脂、纳米二氧化钛、交联剂、稳定剂。制备过程：（1）称取1g壳聚糖完全溶解于浓度为2%的醋酸溶液中，制备壳聚糖醋酸溶液；（2）往壳聚糖醋酸溶液中加入0.25%的纳米二氧化钛和0.5g稳定剂对壳聚糖进行改性，同时在磁力搅拌器转速550r/min条件下反应2h，形成壳聚糖/二氧化钛乳状液；（3）用去离子水制备鼠李糖脂溶液，然后将乳状液与鼠李糖脂溶液以体积比1:1进行混合，再加入0.5%的稳定剂，并且调整PH值为5，在磁力搅拌器转速650r/min、25℃条件下反应24h，合成壳聚糖/二氧化钛/鼠李糖脂三聚络合物。
此方法制备的壳聚糖多维改性抗菌剂的抗菌机理：首先，使用纳米二氧化钛作为改性剂对壳聚糖性能进行改性，不仅增强壳聚糖的吸附性能、显著提高抗菌剂在可见光下的抗菌性能还兼备具有了纳米特性 (小尺寸效应、宏观量子隧道效应等)；其次，壳聚糖和鼠李糖脂结合在一起可以增加表面电荷和改善稳定性，利用带负电荷的三聚磷酸钠与阴离子表面活性剂的排斥静电作用，使壳聚糖的NH3 +基团数高于鼠李糖脂的COO-基团数，壳聚糖混合物纳粒中更多的壳聚糖游离氨基会导致更高的电荷密度，从而增强壳聚糖的抗菌作用。最后，壳聚糖/纳米二氧化钛/鼠李糖脂复合纳米粒对细胞膜的吸附作用导致鼠李糖脂在细菌细胞表面附近的局部释放，这可能有助于鼠李糖脂进入细胞膜，达到有效抗菌。
实施例2：体外抑菌实验（实验菌株：大肠杆菌、金黄色葡萄球菌）
方法一（纸片法）：①培养基的配置：将5g蛋白胨、1.5g牛肉膏、2.5g氯化钠、7.5g琼脂粉加入到500ml蒸馏水中，加热充分溶解后，利用NaOH溶液调节配制的培养基PH，使PH在7.2～7.4左右，于121℃下高温灭菌20min；②在超净工作台上将灭菌后的混合溶液倒入平板中，等待冷却，制成固体培养基；③在超净工作台上用灭菌过的移液枪移取适量菌悬液于培养基中，在酒精灯下进行涂布操作，涂布要均匀、充分；④涂布完成后，用镊子将圆片滤纸在不同的抗菌剂中浸渍片刻，取出置于带菌培养基平板的中央，盖上盖子；⑤把平板倒扣放置在适宜温度下，培养2-3天，观察滤纸片周围抑菌圈的有无及大小。
方法二（琼脂稀释法）：①吸取2ml配制好的各稀释度抗菌剂加入到琼脂培养皿中，倒入冷却后试管中的培养基后迅速摇匀（顺时针三圈逆时针三圈），待凝固，每个稀释法做平行两个平板，再倒三个空白平板；②吸取0.1ml菌悬液加入平板中，并涂匀，留一个空白作为阳性对照组；③计算抑菌率。抑菌率=（对照组菌落数-处理组菌落数）/对照组菌落数*100%。
纸片法的实验结果：
表1 壳聚糖、壳聚糖/鼠李糖脂、壳聚糖/纳米二氧化钛、
壳聚糖/纳米二氧化钛/鼠李糖脂对照组抑菌圈直径表
琼脂稀释法的实验结果：
表2壳聚糖、壳聚糖/鼠李糖脂、壳聚糖/纳米二氧化钛、
壳聚糖/纳米二氧化钛/鼠李糖脂的抑菌率
根据《中华人民共和国行业标准WS/T650-2019》中“抗菌和抑菌效果评价方法”的相关规定为抑菌率≥50%～90%，判定有抑菌作用；抑菌率≥90%，判有较强抑菌作用。本发明所述壳聚糖多维改性抗菌剂的抑菌率≥90%，因此具有较强抑菌作用。
以上所述仅为本发明的优选实施例，并不用于限制本发明，尽管参照前述实施例对本发明进行了详细说明，对于本领域的技术人员来说，其依然可以对前述各实施例所记载的技术方案进行修改，或者对其中部分技术特征进行等同替换。凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。
Claims (9)
1.一种壳聚糖多维改性抗菌剂，其特征在于，该抗菌剂包括壳聚糖、鼠李糖脂、纳米二氧化钛、交联剂、稳定剂。
2.根据权利要求1所述一种壳聚糖多维改性抗菌剂，其特征在于，所述壳聚糖为低分子壳聚糖，分子量在10000以内，脱乙酰度≥80%。
3.根据权利要求1所述一种壳聚糖多维改性抗菌剂，其特征在于，所述鼠李糖脂是一种棕色膏状或白色粉末状的生物表面活性剂，纯度90%以上，具有抗菌功效。
4.根据权利要求1所述一种壳聚糖多维改性抗菌剂，其特征在于，所述纳米二氧化钛属于锐钛型纳米二氧化钛，是一种白色疏松粉末，直径在100纳米以下，具有抗菌、自洁净、抗老化性能。
5.根据权利要求1所述一种壳聚糖多维改性抗菌剂，其特征在于，所述交联剂为三聚磷酸钠、戊二醛、氯化钙等。
6.根据权利要求1所述一种壳聚糖多维改性抗菌剂，其特征在于，所述稳定剂包括有明胶、甘油、聚山梨酯类、山梨醇、聚乙二醇等。
7.根据权利要求1-6任一项所述壳聚糖多维改性抗菌剂的制备方法，依次包括下述步骤：
（1）以纳米二氧化钛作为改性剂加入到壳聚糖醋酸溶液中，然后缓慢滴加适量的交联剂，保持30-40滴/min，同时在磁力搅拌器转速550r/min条件下反应2h，形成壳聚糖/二氧化钛乳状液；
（2）将乳状液与鼠李糖脂溶液以体积比1:1进行混合，再加入0.5%的稳定剂，调整PH值为4～6，在磁力搅拌器转速650r/min、25℃条件下反应24h，合成壳聚糖/二氧化钛/鼠李糖脂三聚络合物。
8.根据权利要求7所述的壳聚糖多维改性抗菌剂制备方法，其特征在于，壳聚糖和鼠李糖脂溶液浓度范围在0.5～3.5mg/ml之间。
9.根据权利要求7所述的壳聚糖多维改性抗菌剂制备方法，其特征在于，壳聚糖混合液中需保持壳聚糖与交联剂的质量比在2:1～6:1范围内。
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