CN114097788B - 一种类芬顿缓释抑菌水凝胶及其制备方法与应用 - Google Patents

一种类芬顿缓释抑菌水凝胶及其制备方法与应用 Download PDF

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CN114097788B
CN114097788B CN202111413303.2A CN202111413303A CN114097788B CN 114097788 B CN114097788 B CN 114097788B CN 202111413303 A CN202111413303 A CN 202111413303A CN 114097788 B CN114097788 B CN 114097788B
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CN114097788A (zh
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蒋刚彪
吴官平
胡甜
杨子明
杨溢玫
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South China Agricultural University
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Abstract

本发明公开了一种类芬顿缓释抑菌水凝胶及其制备方法与应用。本发明以天然高分子材料、过氧化物、纳米金属氧化物、铵盐或弱酸为原料，通过低速搅拌法合成类芬顿缓释抑菌水凝胶。本发明所得的水凝胶生物安全性高，成本低廉、工艺简单，制备条件温和，能长效缓释活性氧和金属离子，通过类芬顿反应能持续长效抑制微生物的正常生命活动，可以作为保鲜剂延长各类食品的货架期，还可以作为创伤愈合敷料防止创口感染，加速创口愈合，广泛应用于生物医学、食品加工等领域。

Description

一种类芬顿缓释抑菌水凝胶及其制备方法与应用
技术领域
本发明属于天然高分子材料制备技术领域，具体涉及一种类芬顿缓释抑菌水凝胶及其制备方法与应用。
背景技术
活性氧是一种绿色氧化剂，实际应用常见于芬顿试剂中。在生物医学领域常用于杀菌消毒和抑制癌细胞生长。因其作用速度快效率高、价格低廉等特点受到人们的青睐。但传统芬顿试剂中的活性氧作用时间短，无法实现长效作用、只能以液态形式存在等问题极大限制了其实际应用范围。由于活性氧极其活泼的性质，故常见的材料无法实现活性氧和具有催化作用的金属离子(以下统称金属离子)两者的长期稳定共存，进而无法实现将活性氧长效缓释应用。
发明内容
为解决现有技术的缺点和不足之处，本发明的首要目的在于提供一种类芬顿缓释抑菌水凝胶的制备方法。本发明以天然高分子材料、过氧化物、纳米金属氧化物、铵盐或弱酸为原料，通过低速搅拌法合成。本发明所得的水凝胶生物安全性高，工艺简单，制备条件温和。能长效缓释活性氧和金属离子，通过类芬顿反应能有效抑制细菌的正常生命活动。本发明所得的水凝胶能满足于在生物医学、食品加工等众多领域中的实际应用。
本发明的另一目的在于提供上述方法制得的一种类芬顿缓释抑菌水凝胶。
本发明的再一目的在于提供上述一种类芬顿缓释抑菌水凝胶在药物制备和食品保鲜中的应用。
本发明所得的水凝胶可以作为食品保鲜剂实现长效抑菌，进而延长各类食品的货架期。
本发明所得的水凝胶可以充当创伤愈合敷料，在创伤愈合过程中实现持续抑菌，防止伤口感染，进而加速创伤愈合。
本发明目的通过以下技术方案实现：
一种类芬顿缓释抑菌水凝胶的制备方法，包括以下步骤：
将天然高分子材料溶于溶剂中，先加入纳米金属氧化物并分散均匀，再加入过氧化物并分散均匀，最后加入铵盐和/或弱酸，搅拌混合，得到类芬顿缓释抑菌水凝胶；
其中天然高分子材料、溶剂、纳米金属氧化物、过氧化物以及铵盐和/或弱酸的比例为0.1～1g：10～25mL：0.1～0.5g：0.01～0.5g：0.01g。
优选的，所述天然高分子材料、溶剂、纳米金属氧化物、过氧化物以及铵盐和/或弱酸的比例为0.3g：20mL：0.25g：0.01～0.5g：0.01g。
优选的，所述天然高分子材料为羧甲基纤维素钠、羧甲基壳聚糖、海藻酸钠、透明质酸钠、琼脂糖和壳聚糖中至少一种。
优选的，所述纳米金属氧化物为纳米氧化锌、纳米氧化钙、纳米氧化镁和纳米氧化铝中至少一种。
优选的，所述过氧化物为过碳酸钠、过氧化脲、过氧化钙和过氧化氢中至少一种。
优选的，所述铵盐为氯化铵、硫酸铵、硝酸铵、过硫酸铵、柠檬酸铵和单宁酸铵中的至少一种。
优选的，所述弱酸为硅酸、氢氰酸、次氯酸、酒石酸、柠檬酸、单宁酸、苹果酸、水杨酸、乙酸和松香中的一种。
优选的，所述溶剂为水、乙醇和二甲基亚砜中的至少一种；更优选为水。
优选的，所述纳米金属氧化物先加入溶剂中并于20～40Hz下超声分散2～3h，再将分散液加入到天然高分子材料溶液中，其中纳米金属氧化物和溶剂的比例为0.1～0.5g：5mL，所述溶剂与高分子溶液中的溶剂相同。
优选的，所述过氧化物加入后在20～40Hz下超声分散2～3h。
优选的，所述搅拌混合的转速为500～1000r/min，时间为2～4h。
上述方法制得的一种类芬顿缓释抑菌水凝胶。
上述一种类芬顿缓释抑菌水凝胶在药物制备和食品保鲜中的应用。
上述一种类芬顿缓释抑菌水凝胶在制备创伤愈合敷料和作为食品保鲜剂中的应用。
本发明通过天然高分子材料实现活性氧和金属离子两者的长期稳定共存。首先，铵盐或弱酸能在一定程度上稳定活性氧。其次，通过天然高分子材料分别包覆活性氧和纳米金属氧化物，高分子与纳米金属氧化物形成水凝胶。最后，高分子与活性氧的氢键作用又能进一步稳定活性氧。如此一来既阻隔了两者，又能在后续实现缓释活性氧和金属离子这一目的。本发明的水凝胶能实现长效缓释活性氧和金属离子，通过类芬顿反应实现长效杀菌消毒的目的，进而拓宽芬顿试剂的应用范围，充分发挥活性氧高效抑菌作用。
与现有技术相比，本发明具有以下优点及有益效果：
(1)本发明所得的水凝胶生物安全性高、成本低廉、工艺简单。
(2)本发明能有效延长活性氧的作用时间，持续长效杀灭微生物。
附图说明
图1为本发明实施例2所得水凝胶在pH＝7.4的PBS(磷酸缓冲盐溶液)中活性氧的缓释情况。
图2为本发明实施例2所得水凝胶在pH＝7.4的PBS(磷酸缓冲盐溶液)中锌离子的缓释情况。
图3为本发明实施例10中对大肠杆菌(Escherichia coli，ATCC 25922，购自中国广东微生物所菌种保藏管理中心)的抑制效果情况，其中标号5为空白组、6为高分子组、7为过碳酸钠加纳米氧化钙组、8为类芬顿缓释水凝胶组。
图4为本发明实施例11中对金黄色葡萄球菌(Staphylococcus aureus，S.aureus，ATCC 6538，购自中国广东微生物所菌种保藏管理中心)的抑制效果情况图，其中标号5为空白组、6为高分子组、7为过碳酸钠加纳米氧化铝组、8为类芬顿缓释水凝胶组。
具体实施方式
下面结合实施例和附图对本发明作进一步详细的描述，但本发明的实施方式不限于此。
本发明实施例中未注明具体条件者，按照常规条件或者制造商建议的条件进行。所用未注明生产厂商者的原料、试剂等，均为可以通过市售购买获得的常规产品。
实施例1
(1)量取8ml水，再加入2ml无水乙醇，将0.1g羧甲基纤维素钠溶于其中。
(2)将0.5g纳米氧化钙超声分散于5ml水中，再将纳米氧化钙分散液加入(1)溶液中。
(3)将0.01g过氧化钙超声分散于(2)溶液中。
(4)将0.01g氯化铵溶于(3)溶液中。
(5)以500r/min的转速搅拌以上溶液，得到一种类芬顿缓释水凝胶。
实施例2
(1)量取8ml水，再加入2ml无水乙醇，将1g海藻酸钠溶于其中。
(2)将0.4g纳米氧化锌超声分散于5ml水中，再将纳米氧化锌分散液加入(1)溶液中。
(3)将0.01g过氧化脲超声分散于(2)溶液中。
(4)将0.01g硫酸铵溶于(3)溶液中。
(5)以500r/min的转速搅拌以上溶液，得到一种类芬顿缓释水凝胶。
实施例3
(1)量取10ml二甲基亚砜，将0.5g壳聚糖溶于其中。
(2)将0.3g纳米氧化铝超声分散于5ml水中，再将纳米氧化铝分散液加入(1)溶液中。
(3)将0.01g过氧化脲超声分散于(2)溶液中。
(4)将0.01g硝酸铵溶于(3)溶液中。
(5)以500r/min的转速搅拌以上溶液，得到一种类芬顿缓释水凝胶。
实施例4
(1)量取10ml水，将0.5g羧甲基壳聚糖溶于其中。
(2)将0.25g纳米氧化锌超声分散于5ml水中，再将纳米氧化锌分散液加入(1)溶液中。
(3)将1ml的3wt％过氧化氢溶液超声分散于(2)溶液中。
(4)将0.01g过硫酸铵溶于(3)溶液中。
(5)以500r/min的转速搅拌以上溶液，得到一种类芬顿缓释水凝胶。
实施例5
(1)量取10ml水，加热至90℃，将0.5g琼脂糖溶于其中，继续加热琼脂糖溶液。
(2)将0.25g纳米氧化镁超声分散于5ml水中，再将纳米金属氧化镁分散液加入(1)溶液中。
(3)将0.5g的过碳酸钠超声分散于(2)溶液中。
(4)将0.01g过硫酸铵溶于(3)溶液中。
(5)以500r/min的转速搅拌以上溶液，冷却后得到一种类芬顿缓释水凝胶。
实施例6
(1)量取10ml水，将0.5g透明质酸钠溶于其中。
(2)将0.25g纳米氧化钙超声分散于5ml水中，再将纳米氧化钙分散液加入(1)溶液中。
(3)将0.5g的过氧化钙超声分散于(2)溶液中。
(4)将0.01g柠檬酸溶于(3)溶液中。
(5)以500r/min的转速搅拌以上溶液，得到一种类芬顿缓释水凝胶。
实施例7
(1)量取10ml水，将0.5g羧甲基壳聚糖溶于其中。
(2)将0.25g纳米氧化铝超声分散于5ml水中，再将纳米氧化铝分散液加入(1)溶液中。
(3)将0.5g的过碳酸钠超声分散于(2)溶液中。
(4)将0.01g酒石酸溶于(3)溶液中。
(5)以500r/min的转速搅拌以上溶液，得到一种类芬顿缓释水凝胶。
实施例8
(1)量取15ml水，将0.8g海藻酸钠溶于其中。
(2)将0.25g纳米氧化镁超声分散于5ml水中，再将纳米氧化镁分散液加入(1)溶液中。
(3)将1ml的3wt％过氧化氢溶液超声分散于(2)溶液中。
(4)将0.01g单宁酸溶于(3)溶液中。
(5)以500r/min的转速搅拌以上溶液，得到一种类芬顿缓释水凝胶。
实施例9
(1)量取15ml水，将0.8g海藻酸钠溶于其中。
(2)将0.25g纳米氧化锌超声分散于5ml水中，再将纳米氧化锌分散液加入(1)溶液中。
(3)将0.3g的过氧化脲超声分散于(2)溶液中。
(4)将0.01g乙酸溶于(3)溶液中。
(5)以500r/min的转速搅拌以上溶液，得到一种类芬顿缓释水凝胶。
实施例10
(1)量取15ml水，将0.8g海藻酸钠溶于其中。
(2)将0.25g纳米氧化钙超声分散于5ml水中，再将纳米氧化钙分散液加入(1)溶液中。
(3)将0.2g的过碳酸钠超声分散于(2)溶液中。
(4)将0.01g硫酸铵溶于(3)溶液中。
(5)以500r/min的转速搅拌以上溶液，得到一种类芬顿缓释水凝胶。
(6)将10g蛋白胨、10g氯化钠、5g牛肉浸膏、15g琼脂和1000ml蒸馏水配成固体培养基。灭菌后趁热倒入培养皿中备用。
(7)将10g蛋白胨、10g氯化钠、5g牛肉浸膏、1000ml蒸馏水配成液体培养基。灭菌后趁热倒入锥形瓶中备用。
(8)将大肠杆菌取100μL别加入100ml液体培养基中，于37℃、160rpm下在摇床中培养18h。
(9)在超净工作台中完成以下操作，于添加了适量固体培养基的培养皿中加入活化后的大肠杆菌，用三角涂布器以由外向里的圆周涂布方式将菌液均匀涂布在平板上。
(10)将培养皿划分为4个区域，并编号5、6、7、8，在培养皿中依次放入4种圆形纸片，放置于30℃恒温培养箱培养18h。4种圆形纸片分别是：空白圆形纸片，于5wt％海藻酸钠溶液中浸润了1h的圆形纸片，于5wt％纳米氧化钙分散液和4wt％过碳酸钠溶液中分别浸润了30min的圆形纸片，浸润了1h(5)凝胶的圆形纸片。
(11)观察并拍照记录抑菌圈情况。
实施例11
(1)量取15ml水，将0.8g海藻酸钠溶于其中。
(2)将0.25g纳米氧化铝超声分散于5ml水中，再将纳米氧化铝分散液加入(1)溶液中。
(3)将0.5g的过碳酸钠超声分散于(2)溶液中。
(4)将0.01g松香溶于乙醇后再加入(3)溶液中。
(5)以500r/min的转速搅拌以上溶液，得到一种类芬顿缓释水凝胶。
(6)将10g蛋白胨、10g氯化钠、5g牛肉浸膏、15g琼脂、1000ml蒸馏水配成固体培养基。灭菌后趁热倒入培养皿中备用。
(7)将10g蛋白胨、10g氯化钠、5g牛肉浸膏、1000ml蒸馏水配成液体培养基。灭菌后趁热倒入锥形瓶中备用。
(8)将金黄色葡萄球菌取100μL分别加入100ml液体培养基中，于37℃、160rpm下在摇床中培养18h。
(9)在超净工作台中完成以下操作，于添加了适量固体培养基的培养皿中加入活化后的金黄色葡萄球菌，用三角涂布器以由外向里的圆周涂布方式将菌液均匀涂布在平板上。
(10)将培养皿划分为4个区域，并编号5、6、7、8，在培养皿中依次放入4种圆形纸片，放置于30℃恒温培养箱培养18h。4种圆形纸片分别是：空白圆形纸片，于5wt％海藻酸钠溶液中浸润了1h的圆形纸片，于5wt％纳米氧化铝分散液和4wt％过碳酸钠溶液中分别浸润了30min的圆形纸片，浸润了1h(5)凝胶的圆形纸片。
(11)观察并拍照记录抑菌圈情况。
实施例12
(1)量取20ml水，将0.3g羧甲基纤维素钠溶于其中。
(2)将0.25g纳米氧化锌超声分散于5ml水中，再将纳米氧化锌分散液加入(1)溶液中。
(3)将1ml的3wt％过氧化氢溶液超声分散于(2)溶液中。
(4)将0.01g硝酸铵溶于(3)溶液中。
(5)以500r/min的转速搅拌以上溶液，得到一种类芬顿缓释水凝胶。
(6)将以上一种类芬顿缓释水凝胶取1g置于饼干底部，再进行包装。
实施例13
(1)量取20ml水，将0.3g羧甲基纤维素钠溶于其中。
(2)将0.25g纳米氧化钙超声分散于5ml水中，再将纳米氧化钙分散液加入(1)溶液中。
(3)将0.01g过碳酸钠超声分散于(2)溶液中。
(4)将0.01g氯化铵溶于(3)溶液中。
(5)以500r/min的转速搅拌以上溶液，得到一种类芬顿缓释水凝胶。
(6)将以上一种类芬顿缓释水凝胶取1g置于蛋糕底部，再进行包装。
实施例14
(1)量取20ml水，将0.3g羧甲基纤维素钠溶于其中。
(2)将0.25g纳米氧化铝超声分散于5ml水中，再将纳米氧化铝分散液加入(1)溶液中。
(3)将0.5g过氧化脲超声分散于(2)溶液中。
(4)将0.01g氯化铵溶于(3)溶液中。
(5)以500r/min的转速搅拌以上溶液，得到一种类芬顿缓释水凝胶。
(6)将以上一种类芬顿缓释水凝胶取1g置于面包表面，再进行包装。
实施例15
(1)量取20ml水，将0.3g羧甲基纤维素钠溶于其中。
(2)将0.25g纳米氧化镁超声分散于5ml水中，再将纳米氧化镁分散液加入(1)溶液中。
(3)将0.1g过氧化脲超声分散于(2)溶液中。
(4)将0.01g氯化铵溶于(3)溶液中。
(5)以500r/min的转速搅拌以上溶液，得到一种类芬顿缓释水凝胶。
(6)将以上一种类芬顿缓释水凝胶均匀覆盖于皮肤创口表面。
上述实施例为本发明较佳的实施方式，但本发明的实施方式并不受上述实施例的限制，其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化，均应为等效的置换方式，都包含在本发明的保护范围之内。

Claims (8)

1.一种类芬顿缓释抑菌水凝胶的制备方法，其特征在于，包括以下步骤：
将天然高分子材料溶于溶剂中，先加入纳米金属氧化物并分散均匀，再加入过氧化物并分散均匀，最后加入铵盐和/或弱酸，搅拌混合，得到类芬顿缓释抑菌水凝胶；
其中天然高分子材料、溶剂、纳米金属氧化物、过氧化物以及铵盐和/或弱酸的比例为0.1～1g：10～25mL：0.1～0.5g：0.01～0.5g：0.01g；
所述铵盐为氯化铵、硫酸铵、硝酸铵、过硫酸铵、柠檬酸铵和单宁酸铵中的至少一种；
所述弱酸为硅酸、氢氰酸、次氯酸、酒石酸、柠檬酸、单宁酸、苹果酸、水杨酸、乙酸和松香中的一种；
所述过氧化物为过碳酸钠、过氧化脲、过氧化钙和过氧化氢中至少一种。
2.根据权利要求1所述一种类芬顿缓释抑菌水凝胶的制备方法，其特征在于，所述天然高分子材料、溶剂、纳米金属氧化物、过氧化物以及铵盐和/或弱酸的比例为0.3g：20mL：0.25g：0.01～0.5g：0.01g。
3.根据权利要求1所述一种类芬顿缓释抑菌水凝胶的制备方法，其特征在于，所述天然高分子材料为羧甲基纤维素钠、羧甲基壳聚糖、海藻酸钠、透明质酸钠、琼脂糖和壳聚糖中至少一种；
所述纳米金属氧化物为纳米氧化锌、纳米氧化钙、纳米氧化镁和纳米氧化铝中至少一种。
4.根据权利要求1所述一种类芬顿缓释抑菌水凝胶的制备方法，其特征在于，所述纳米金属氧化物先加入溶剂中并于20～40Hz下超声分散2～3h，再将分散液加入到天然高分子材料溶液中，其中纳米金属氧化物和溶剂的比例为0.1～0.5g：5mL。
5.根据权利要求1所述一种类芬顿缓释抑菌水凝胶的制备方法，其特征在于，所述过氧化物加入后在20～40Hz下超声分散2～3h；所述搅拌混合的转速为500～1000r/min，时间为2～4h。
6.根据权利要求1或4所述一种类芬顿缓释抑菌水凝胶的制备方法，其特征在于，所述溶剂为水、乙醇和二甲基亚砜中的至少一种。
7.权利要求1～6任一项所述方法制得的一种类芬顿缓释抑菌水凝胶。
8.权利要求7所述一种类芬顿缓释抑菌水凝胶在药物制备和食品保鲜中的应用。
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