CN108950877A - 致病菌诱捕过滤抗菌膜制备方法 - Google Patents

致病菌诱捕过滤抗菌膜制备方法 Download PDF

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CN108950877A
CN108950877A CN201810964110.8A CN201810964110A CN108950877A CN 108950877 A CN108950877 A CN 108950877A CN 201810964110 A CN201810964110 A CN 201810964110A CN 108950877 A CN108950877 A CN 108950877A
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杨维巧
蒋佳男
李喜宏
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Zhong Xi (tianjin) Jujube Industry Technical Engineering Center
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Abstract

本发明涉及一种致病菌诱捕过滤抗菌膜制备方法,步骤如下:包括:聚乳酸溶液的配制、Fe3O4磁性纳米粒子制备、复合流体的制备、静电纺丝、纤维膜性能表征。本发明使用聚乳酸作为成纤基体,依据病原菌趋磁特性,以共沉淀法制备Fe3O4磁性粒子,利用静电纺丝制备技术,将聚乳酸与磁性粒子结合,形成低阻力、高渗透性的多孔载体界面,同时宏观颗粒将转为超微纳米粒子,充分发挥纳米多孔界面与小尺寸过滤双向增强的优势,能有效的截留大部分微生物和部分病毒。将高分子材料学与食品科学相结合,开发新型多孔致病菌诱捕过滤抗菌膜填补我国纳米级过滤膜制备的空白。

Description

致病菌诱捕过滤抗菌膜制备方法
技术领域
本发明属于食品加工领域,具体涉及一种致病菌诱捕过滤抗菌膜制备方法。
背景技术
果汁中含有大量的果胶、鞣质、纤维素、蛋白质、淀粉等大分子,这些物质在汁液中进行缓慢的物理变化和化学反应,导致果汁在加工、贮藏、销售期间变质,直接影响果汁品质。现阶段常采用明胶、酶解、砂滤棒过滤等方法澄清果汁,此外,传统果汁加工中均需增加额外的杀菌工艺,以达到长期保鲜的效果,但是设备投资大,工艺繁琐。
目前,采用过滤膜澄清果汁是国内外较为先进的澄清工艺,中国专利CN107335278A公开了一种果汁过滤膜,包括金属滤层、过滤层、支撑层,金属滤层由三层倾斜夹缝的金属子层构成,过滤层由多孔聚酯纤维层组成,支撑层由金属网构成,通过金属子层不同的倾斜角度使果汁通过滤膜时能够高效过滤杂质;中国专利CN204377839U公开了一种果汁超滤设备,采用带有折叠状的过滤膜的滤芯对果汁进行过滤,达到了清汁出汁透过率达到95%的澄清效果。滤芯和过滤桶体之间采用可拆卸连接,方便清洗,将过滤膜中过滤的杂质冲洗干净,该滤芯可反复使用,节省成本。以上果汁过滤的方法各有其优点,如可回收利用,过滤杂质效果好,便于清洗更换等,但过滤时间长,能耗大,无法达到杀菌效果。
本发明将聚乳酸与Fe3O4共混利用静电纺丝技术制备新型致病菌诱捕过滤材料。采用静电纺丝(电纺)方法构建磁性粒子包埋其中的孔径介于纳米与微米孔径的微孔三维网状结构,具有更加独特的低阻力、高渗透的微孔吸附特性,利用微生物趋磁吸附作用,使病原菌由传统的被动接触杀菌变为主动诱捕策略。磁性纳米粒子多孔纤维载体界面,将澄清工艺与磁性抗菌粒子合二为一,极大程度简化澄清型果汁饮料的制备工艺,可以实现磁性吸引与高效抗菌的创新结合。
聚乳酸(PLA)是一种新型的生物基及可生物降解材料,因其具有良好的生物可降解性,是公认的环境友好材料。此外热稳定性强,生物相容性、光泽度、透明性和耐热性好,还具有一定的耐菌性、阻燃性,加之与磁性Fe3O4粒子相结合。在继承前人制备的果汁过滤膜优点的基础上,克服了普通澄清型果汁过滤膜的缺点,不仅可以高效的过滤澄清型果汁中的各种大分子杂质,还能达到过滤抗菌的效果,操作简单,过滤时间短,成本低耗能少,且此类过滤膜目前还未有报道。
发明内容
本发明提的目的在克服现有技术的不足之处,提供一种保鲜效果好、保鲜期长的致病菌诱捕过滤抗菌膜制备方法。
本发明实现目的的技术方案如下:
一种致病菌诱捕过滤抗菌膜的制备方法,包括以下步骤:
⑴聚乳酸溶液的配制:
选定左旋聚乳酸(L-PLA,Mw=2.7×105kDa)作为成纤基材,将5.0g的聚乳酸粉末加入约20%的N,N-二甲基甲酰胺(DMF)之中,在65℃水浴条件下使用磁力搅拌器搅拌2小时,形成聚乳酸质量分数为8%-10%的均匀诱明的聚乳酸溶液;
⑵Fe3O4纳米粒子的制备方法:
本发明采用共沉淀法,即利用Fe2++2Fe3++8OH-→Fe3O4↓+4H2O反应,制备Fe3O4纳米磁性粒子。将二价铁盐和三价铁盐的混合溶液加入到三口烧瓶中,滴液漏斗中加入沉淀剂NH3·H2O,在氮气气氛下将氨水加到反应体系中,使体系的pH≥10,恒温水浴,匀速搅拌30min后结束反应,用蒸馏水反复洗涤直至中性,倾去上层清液,60℃下真空干燥后,研磨即得纳米Fe3O4粒子;
⑶Fe3O4/聚乳酸复合流体的制备方法:
称取制备的Fe3O4纳米粒子溶于约20%的DMF后,将聚乳酸溶液快速倒入含Fe3O4纳米粒子的溶液中,30℃下匀速搅拌10小时,在搅拌器搅拌过程中溶解均匀,即成Fe3O4/聚乳酸复合流体;
⑷静电纺丝技术制备致病菌诱捕过滤抗菌膜:
静电纺丝装置为YFSP-T(M3T2QG),10mL注射器安装在静电纺丝装置卡槽处,电磁线圈与气泵双重调控进样流速,精准调控进样速率。选用不粘型铝箔纸能达到将纤维膜更好分离铝箔的效果,实验中滚筒式收集器控制在80r/min,推柱速率为1mm/min,电压13-18kv,进样针至收集器垂直距离为10-20cm,连接注射器的不锈钢针头外观尺寸为0.7mm×32mm,直径0.390mm与注射器相连,在不锈钢针头处连接高压电场为实验提供高压电源,静电纺丝装置环境内温度20℃,环境相对湿度RH为22%,通风晾干后从铝箔纸上取下纳米纤维膜。
而且,所述二价铁盐为FeSO4·7H2O,所述三价铁盐为FeCl3·6H2O。
本发明与现有技术相比的优点和积极效果如下:
1、本发明首次研发静电纺丝聚乳酸多孔磁性纳米纤维膜,具有高孔隙率、孔径均一、形貌可控的三维多孔连通结构特点,这一100-200nm直径纤维组成的多孔连通结构能够过滤果汁中的各种大分子杂质和大部分微生物,极细的纳米纤维增大了杂质与膜接触的表面积,加速过滤果汁中的杂质。
2、本发明利用病原菌主动趋磁构效特性和聚乳酸磁性纳米纤维多孔界面与小尺寸效应,使环境内病原菌由于磁性吸引加速吸附到多孔纳米纤维膜上,将以膜两侧形成蒸汽压差为传质动力,为Fe3O4磁性粒子诱捕果汁中病原菌将提供有效吸附的渗透载体,提高传输速率,实现磁性主动吸引病原菌与聚乳酸超微多孔纤维结构高效过滤创新结合。
3、本发明以澄清型果汁病原微生物为目标,明确多孔纳米纤维低渗透阻力与超大比表面积特性过滤效果,实现澄清型果汁在不做任何杀菌处理情况下,通过致病菌诱捕过滤抗菌膜,达到高效过滤病菌效果,开发新型多孔致病菌诱捕过滤抗菌膜填补我国纳米级磁性抗菌纤维膜制备的空白。
具体实施方式
下面通过具体实施例对本申请作进一步详述,以下实施例只是描述性的,不是限定性的,不能依次限定本申请的保护范围。
一种致病菌诱捕过滤抗菌膜制备方法,采用共沉淀法利用Fe2++2Fe3++8OH-→Fe3O4↓+4H2O原理制备Fe3O4纳米磁性粒子,然后通过共混包埋法技术制备Fe3O4/聚乳酸复合流体,通过静电纺丝技术,设置特定的纺丝参数,制成纤维直径100-200nm的多孔纳米过滤膜,提高聚乳酸纳米粒子的稳定性,达到高效过滤病菌的目的。
本发明通过以下技术方案实施:
纳米纤维过滤膜的制备方法,包括以下步骤:
⑴聚乳酸的选择和溶液的配制:
选定左旋聚乳酸(L-PLA,Mw=2.7×105kDa)作为成纤基材,将5.0g的聚乳酸粉末加入约20%的N,N-二甲基甲酰胺(DMF)之中,在65℃水浴条件下使用磁力搅拌器搅拌2小时,形成聚乳酸质量分数为8%-10%的均匀诱明的聚乳酸溶液;
⑵Fe3O4的制备方法:
本发明采用共沉淀法,即利用Fe2++2Fe3++8OH-→Fe3O4↓+4H2O反应,制备Fe3O4纳米磁性粒子。将二价铁盐和三价铁盐的混合溶液加入到三口烧瓶中,滴液漏斗中加入沉淀剂NH3·H2O,在氮气气氛下将氨水加到反应体系中,使体系的pH≥10,恒温水浴,匀速搅拌30min后结束反应,用蒸馏水反复洗涤直至中性,倾去上层清液,60℃下真空干燥后,研磨即得纳米Fe3O4粒子;
二价铁盐(FeSO4·7H2O)和三价铁盐(FeCl3·6H2O)的使用浓度为1:2
⑶Fe3O4/聚乳酸复合流体的制备方法:
共混包埋法是指将磁性超微颗粒均匀地分散在高分子溶液中,通过范德华力、氢键、配位键和共价键等作用将高分子物质缠绕在无机磁性颗粒表面,形成均质、稳定的复合流体。本发明将准确称取Fe3O4纳米粒子,溶于约20%的DMF后,将聚乳酸溶液快速倒入含Fe3O4纳米粒子的溶液中,在搅拌器搅拌过程中溶解均匀,即成Fe3O4/聚乳酸复合流体。
⑷静电纺丝技术制备致病菌诱捕过滤抗菌膜:
静电纺丝装置为YFSP-T(M3T2QG),10mL注射器安装在静电纺丝装置卡槽处,电磁线圈与气泵双重调控进样流速,精准调控进样速率。选用不粘型铝箔纸能达到将纤维膜更好分离铝箔的效果,实验中滚筒式收集器控制在80r/min,推柱速率为1mm/min,电压13-18kv,进样针至收集器垂直距离为10-20cm,连接注射器的不锈钢针头外观尺寸为0.7mm×32mm,直径0.390mm与注射器相连,在不锈钢针头处连接高压电场为实验提供高压电源,静电纺丝装置环境内温度20℃,环境相对湿度RH为22%,连续静电纺丝10-12h,通风晾干后从铝箔纸上取下纳米纤维膜。
本发明中纳米磁性纤维膜对大肠杆菌、金黄色葡萄球菌的抗性表达的是抑菌圈的直径。
表1致病菌诱捕过滤抗菌膜的抑菌效果
表2 Fe3O4矫顽力与磁性强度
样品 矫顽力(Oe) 饱和磁强度(emu/g) 剩余磁强度(emu/g)
Fe3O4 27.38 1.92 34.30
1.Fe3O4的制备原理
采用共沉淀法,即利用Fe2++2Fe3++8OH-→Fe3O4↓+4H2O反应,制备Fe3O4纳米磁性粒子,再采用共混包埋法技术制备Fe3O4/壳聚糖纺丝液。通过分析壳聚糖流体性质可知,特定溶剂溶解后的壳聚糖流体带正电荷,能够通过静电或氢键为主导,实现对磁性Fe3O4粒子包埋形成稳定的分散胶体悬浮液,从而使复合体系呈现均匀分散体系。
2.协同抗菌分子机制
细菌体内铁载体达500多种,其中异羟肟酸型铁载体在大肠杆菌中的吸收是迄今为止研究表征较好的细菌吸铁系统,特异性细菌自身周质空间转运蛋白FhuD可以绑定和穿梭各种异羟肟酸型铁载体,是初级主动转运蛋白转运系统重要组成。超顺磁Fe3O4强磁场与铁载体靶向吸附将介导铁载体与特异性受体蛋白结合反应,一方面,极大程度引起FhuD蛋白二级结构域构象的重大变化。另一方面,铁载体参与的Fe3+摄取过程很可能将“搬运”的Fe3+作为底物,直接参与ROS自由基的生成反应,产生更多活性氧自由基O2 ,造成细胞膜过氧化伤害,进而影响细胞膜完整性、流动性等细胞超微结构的变化。

Claims (2)

1.一种致病菌诱捕过滤抗菌膜的制备方法,其特征在于:包括以下步骤:
⑴聚乳酸溶液的配制:
选定左旋聚乳酸作为成纤基材,将聚乳酸粉末加入20wt%的N,N-二甲基甲酰胺之中,在65℃水浴条件下使用磁力搅拌器搅拌2-2.5小时,形成聚乳酸质量分数为8%-10%的均匀诱明的聚乳酸溶液;
⑵Fe3O4纳米粒子的制备方法:
本发明采用共沉淀法,即利用Fe2++2Fe3++8OH-→Fe3O4↓+4H2O反应,制备Fe3O4纳米磁性粒子。将二价铁盐和三价铁盐的混合溶液加入到容器中,滴液漏斗中加入沉淀剂NH3·H2O,在氮气气氛下将氨水加到反应体系中,使体系的pH≥10,恒温水浴,匀速搅拌30min后结束反应,用蒸馏水反复洗涤直至中性,倾去上层清液,60℃下真空干燥后,研磨即得纳米Fe3O4粒子;
⑶Fe3O4/聚乳酸复合流体的制备方法:
称取制备的Fe3O4纳米粒子溶于约20wt%的DMF后,将聚乳酸溶液快速倒入含Fe3O4纳米粒子的溶液中,30℃下匀速搅拌10小时,在搅拌器搅拌过程中溶解均匀,即成Fe3O4/聚乳酸复合流体;
⑷静电纺丝技术制备致病菌诱捕过滤抗菌膜:
静电纺丝装置为YFSP-T,10mL注射器安装在静电纺丝装置卡槽处,电磁线圈与气泵双重调控进样流速,精准调控进样速率,选用不粘型铝箔纸能达到将纤维膜更好分离铝箔的效果,实验中滚筒式收集器控制在80r/min,推柱速率为1mm/min,电压13-18kv,进样针至收集器垂直距离为10-20cm,连接注射器的不锈钢针头外观尺寸为0.7mm×32mm,直径0.390mm与注射器相连,在不锈钢针头处连接高压电场为实验提供高压电源,静电纺丝装置环境内温度20℃,环境相对湿度RH为22%,通风晾干后从铝箔纸上取下纳米纤维膜。
2.根据权利要求1所述的致病菌诱捕过滤抗菌膜的生产方法,其特征在于:所述二价铁盐为FeSO4·7H2O,所述三价铁盐为FeCl3·6H2O。
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