CN113273605A - 壳聚糖-银配合物抗菌凝胶及其制备方法和保鲜应用 - Google Patents
壳聚糖-银配合物抗菌凝胶及其制备方法和保鲜应用 Download PDFInfo
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Abstract
本发明为壳聚糖‑银配合物抗菌凝胶及其制备方法和保鲜应用，提供一种具有优良抗菌性的壳聚糖‑银水凝胶及其制备方法和应用。这种复合水凝胶的制备方法包括：硝酸银和壳聚糖在超声的作用下发生共组装，形成了澄清透明的配合物水凝胶。该水凝胶中，银离子与壳聚糖的协同作用大大提高了本发明的抗菌保鲜性。本发明具有抗菌、保鲜、易涂抹等性能；制备方法简单，成本低廉，可商品化，抑菌效果好，稳定性好等优点；制备过程中没有添加任何还原剂与分散剂，绿色环保。本发明的壳聚糖‑银水凝胶各组分通过协同作用，使抗菌性显著提高，在葡萄保鲜领域有广阔的应用前景。
Description
技术领域
本发明属于食品保鲜技术领域，特别是涉及一种用于葡萄保鲜的壳聚糖-银抗菌凝胶及其制备方法和应用。
背景技术
葡萄由于表皮脆弱且含水量较大，在贮运时容易破损，导致微生物的侵染繁殖而腐烂变质，严重影响葡萄保鲜品质和经济效益。据统计，我国目前由于葡萄贮藏保鲜处理不好，致使葡萄每年贮藏损耗率在20％～25％，物流损失率高达40％，远低于发达国家葡萄物流损失率。葡萄的贮运保鲜问题日趋突出，如何延长葡萄的贮藏时间，保持其品质不变，抑制微生物的侵害，已成为亟待解决的问题。
传统的化学食品防腐保鲜剂因其对人体健康有影响，以及污染环境等弊端已逐渐被取代。选择天然的、绿色安全的保鲜剂受到广泛关注。壳聚糖(chitosan，简称CS)及其衍生物作为一种天然生物大分子，因其具有良好的广谱抑菌性，保湿性且无任何毒副作用走进了人们的视线。但是，单纯的壳聚糖抑菌谱较窄，抑菌效果不是很显著，且吸附性一般涂膜效率低，限制其进一步应用。银及其化合物是最常见的抗菌剂之一，具有广谱抗菌性。但是银离子易被空气氧化而影响抑菌效果，且摄入过多可能会对人体肝脏产生危害。因此，如何协调两种天然抗菌保鲜剂，提高应用效果，对于抑菌保鲜领域应用具有重要意义。
多糖类水凝胶主要是亲水性高分子网络体系，含水量极大(一般其重量90％以上都为水分子)，具有优良的生物相容性与生物降解能力。封装在水凝胶中的物质可被3D网状纤维紧密固定，同时又可隔绝空气从而避免封装物氧化，在固定化酶、细胞分离、蛋白制备和缓释药物等研究中具有重要的作用。
利用上述原理，我们成功制备出具有双重抑菌、保鲜性能的壳聚糖-银配合物水凝胶。本发明制备过程操作简便，成本低，适于大规模生产。实验结果表明，该壳聚糖-银配合物水凝胶有望在抗菌剂与保鲜方面的应用等领域有很好的应用。
发明内容
本发明的目的是提供一种具有优良抗菌保鲜性的壳聚糖-银配合物水凝胶的制备方法及其应用。本发明的壳聚糖-银凝胶是澄清透明的水凝胶，具有可涂抹的性能，可以增强葡萄对细菌的抵抗能力，达到长久的保鲜作用。
本发明中的抗菌性的壳聚糖-银配合物凝胶的制备，包括以下步骤：
(1)配制一定浓度的壳聚糖储备溶液；
(2)配制一定浓度的硝酸银储备溶液；
(3)将一定量的各组分储备溶液混合摇匀，超声即得所述的壳聚糖-银配合物水凝胶。
上述的制备方法步骤(1)中，所述的壳聚糖储备溶液的溶剂为超纯水，壳聚糖储备溶液的浓度为5～20 mg/mL，并加入体积比为1％冰乙酸促进壳聚糖的溶解。
上述的制备方法步骤(2)中，所述的硝酸银储备溶液的溶剂为超纯水，硝酸银储备溶液的浓度为0.3 mol/L，配制硝酸银储备溶液的过程中避光处理。
上述的制备方法步骤(3)中，取100μL的硝酸银储备溶液，加入2.0mL避光处理的螺口瓶中迅速摇匀，再加入800μL的壳聚糖储备溶液，100μL的氢氧化钠溶液(0.2mol/L)配成1.0mL的混合溶液，迅速振荡摇匀，壳聚糖-银配合物水凝胶即可形成。
本发明使用的壳聚糖-银凝胶是通过非共价键自组装得到的，该水凝胶摇动即可迅速形成凝胶。另外，壳聚糖与银粒子结合，增强了水凝胶的机抗菌性与保鲜性。
有益效果
(1)本发明制备的壳聚糖-银配合物凝胶，生物相容性好，易降解，无细胞毒性。
(2)本发明制备的壳聚糖-银配合物凝胶是澄清透明的水凝胶，易涂抹，保湿性好。
(3)本发明加入的壳聚糖与银离子，各自具有广谱抗菌性，两者结合抑菌性能明显提高。
(4)本发明制备方法简单，成本低廉，可商品化，适于大规模生产。
(5)本发明制备壳聚糖-银配合物凝胶有良好的抗菌性和保鲜性，将会在水果保鲜领域有很好的应用前景。
附图说明
图1为壳聚糖-银配合物水凝胶成胶浓度：
图2本发明实施例1中壳聚糖-银配合物紫外光谱图；
图3为壳聚糖-银配合物水凝胶的扫描电子显微镜(SEM)图；
图4壳聚糖-银配合物对青霉菌生长状况影响的数码照片图；
图5壳聚糖-银配合物对黑霉菌生长状况影响的数码照片图。
具体实施方式
为了便于理解本发明，下文将结合说明书附图和具体的实施例对本发明作更全面细致地描述，但本发明的保护范围并不限于以下具体的实施例。
实施例1
称取2.046g的硝酸银于棕色避光的离心管中，加入20mL的超纯水将其溶解，用锡箔纸避光处理的 50mL棕色试剂中，配成0.6mol/L的硝酸银水溶液作为硝酸银储备溶液；称取500mg的壳聚糖，加入 100mL的超纯水将其溶解，再加入体积比为1％的冰乙酸以促进壳聚糖的溶解，配成5mg/mL的壳聚糖储备溶液。称取160mg的氢氧化钠，加入20mL的超纯水将其溶解，配成0.2mol/mL的氢氧化钠储备溶液。
取100μL的硝酸银储备溶液，加入2.0mL避光处理的螺口瓶中迅速摇匀，再加入800μL的壳聚糖储备溶液和100μL的氢氧化钠溶液，配成2.0mL的混合溶液，迅速振荡摇匀，即可形成澄清透明的壳聚糖-银水凝胶。
抗菌实验步骤：称取46gPDA培养基粉末于烧杯中，向烧杯中加入超纯水溶解，用玻璃棒搅拌促进溶解，用超纯水定容至1L的锥形瓶中制备PDA培养基。
取适量自然腐烂的红提葡萄，加入含有100mL无菌水的烧杯中并持续搅拌，取出葡萄，过滤并收集含有真菌孢子的液体。将此滤液视为孢子悬浮液母液，使用梯度稀释法将母液稀释为10-1，10-2，10-3， 10-4，10-5，10-6和10-7等的稀释液。将每种稀释液(1mL)加入到培养皿中，与PDA培养基混合均匀后，待基底凝固后翻转过来置于28℃下培养7d。
培养7d后，选择已经将每个菌落完全分离开的培养皿，在其中选择含有较多数量的同一种类的菌落，用无菌接种环刮取该菌落表面菌丝，将菌丝接种在含有PDA培养基的培养皿上，分别加入不同浓度壳聚糖 -银凝胶均匀涂布在PDA培养基上做抑菌实验。在28℃下培养7d。拍照观察并记录。
上述所有无菌操作，均在超净工作台中进行，所有用到的无菌器皿和物质均经过121℃下30min的高压灭菌处理。
实施例2
取上述实施例1中的硝酸银储备溶液100μL，加入至洗净吹干避光处理的2.0mL螺口瓶中，快速加入800μL与实施例1配制方法相同的壳聚糖储备溶液，迅速摇匀，再加入100μL与实施例1配制方法相同的和100μL氢氧化钠储备溶液，迅速振荡摇匀。即可形成澄清透明的壳聚糖-银水凝胶。
葡萄保鲜实验步骤：将新鲜葡萄分装为四份，分别用浓度为6.0％的壳聚糖-银溶胶处理葡萄，同质量浓度的壳聚糖溶液，同质量浓度的银溶液处理，以不进行处理为CK组。每天分别测定失重率、腐烂率、可溶性固形物、可滴定酸、维生素C含量等指标，并拍照记录。
以上显示和描述了本发明的制备过程、主要特征和本发明的应用。本行业的技术人员应该了解，本发明不受上述实施例的限制。本发明要求保护范围由所附的权利要求书界定。
Claims (4)
1.一种抗菌性的配合物水凝胶的制备方法，其特征在于，包括以下步骤：
用超纯水避光溶解硝酸银晶体，配成硝酸银储备溶液；
用超纯水溶解壳聚糖粉末，加入冰乙酸以促进壳聚糖的溶解，配成壳聚糖储备溶液；
取100μL的硝酸银储备溶液，再加入800μL的壳聚糖储备溶液和100μL的氢氧化钠溶液，配成1.0mL的混合溶液，迅速振荡摇匀，壳聚糖-银水凝胶即可形成。
2.根据权利要求1所述的制备方法，其特征在于，所述硝酸银储备溶液的浓度为0.6mol/L，壳聚糖储备溶液的浓度为5mg/mL，加入体积比为1％冰乙酸。
3.一种如权利要求1所述的制备方法制备得到的复合水凝胶，该壳聚糖-银水凝胶是壳聚糖与银离子发生配位，通过非共价键自组装得到的澄清透明的水凝胶，具有可涂抹的性能。
4.一种由权利要求1所述制备方法合成的壳聚糖-银水凝胶在葡萄抑菌保鲜方面的应用。
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