CN115073768B - 一种负载功能成分双网络水凝胶的制备方法 - Google Patents
一种负载功能成分双网络水凝胶的制备方法 Download PDFInfo
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Abstract
本发明公开了一种负载功能成分双网络水凝胶的制备方法,该方法包括:将乳清蛋白溶于超纯水,在低pH和加热条件下制备乳清蛋白纳米纤维。将乳清蛋白纳米纤维溶液调节为弱酸性,将海藻酸钠、葡萄糖酸δ‑内酯(GDL)、碳酸钙纳米颗粒溶解在弱酸性的乳清蛋白纳米纤维溶液中进行磁力搅拌和超声脱气泡。本发明使用高深宽比填料的方法来提高水凝胶机械强度,在室温下制备得到海藻酸钠/乳清蛋白纳米纤维双网络水凝胶。本发明不仅制备过程简便,而且海藻酸钠/乳清蛋白纳米纤维双网络水凝胶具有较高的凝胶强度和机械性能等显著等优点,并且可以应用于热敏营养物的递送***中。
Description
技术领域
本发明涉及双网络水凝胶制备技术领域,尤其涉及一种海藻酸钠/乳清蛋白纳米纤维的双网络水凝胶的制备方法。
背景技术
水凝胶是一种聚合材料,可以在其结构内保留大量水或生物液体,是一种不会溶解的三维交联网络结构。由多糖和蛋白质制备的水凝胶机械性能较差,由人工合成的水凝胶较天然水凝胶机械性能好,但仍比较脆弱,不能达到理想状态。为了克服单网络水凝胶表现的局限性,目前研究人员已采用经典法、分子支架法、一锅法等制备了双网络水凝胶,由两个独特的互穿聚合物网络组成显著提高机械强度。
在制备水凝胶使用的原料中,蛋白质和多糖因其可生物降解性、来源广泛性和无毒特性而特别有吸引力。基于蛋白质的水凝胶,如乳清蛋白凝胶,由于其高营养价值、优异的功能特性和两亲性,已成为构建混合水凝胶的热门选择,但蛋白质水凝胶会被胃部酸性条件和胃蛋白酶破坏,作为营养载体运输效率差;被称为膳食纤维的多糖如海藻酸钠人体较难消化,但可以被肠道微生物降解,可以保护蛋白质免受胃部影响。与蛋白质作为唯一原料的水凝胶相比,多糖/蛋白质双网络水凝胶有更高的机械强度,更高的运输效率。
大多数多糖-蛋白质形成的双网络水凝胶需要经过热处理,这样不利于包埋姜黄素等热敏性物质。通过加入Ca2+、酶、酸等可以不用经过热处理就形成双网络水凝胶,可以扩大其应用范围。
发明内容
本发明目的旨在提供了一种海藻酸钠-乳清蛋白纳米纤维双网络水凝胶的制备方法(即负载功能成分双网络水凝胶的制备方法),该方法不仅制备过程耗时短、效率高,而且获得的双网络水凝胶具有机械强度高的性质。由粒径较小的纳米碳酸钙代替大分子碳酸钙或者氯化钙,葡萄糖酸δ-内酯溶解过程中由氢离子分解碳酸钙纳米颗粒释放的Ca2+离子,形成的水凝胶均匀性更好,同时具有更好的缓释效果。该水凝胶可以用作热敏营养物质的运输载体,同时拓宽其应用范围,提高营养物的生物利用度。
具体技术方案如下:
一种双网络水凝胶,该双网络水凝胶由海藻酸钠和乳清蛋白纳米纤维制备而成。
一种海藻酸钠/乳清蛋白纳米纤维双网络水凝胶的制备方法(即负载功能成分双网络水凝胶的制备方法),包括以下步骤:
(1)将乳清蛋白溶于超纯水中,在pH为1.5~2.5(最优选为2)的条件下加热搅拌一段时间,调节溶液pH,得到溶液I;
(2)滴加或不滴加热敏性物质到步骤(1)的溶液I中,将混合物放置于室温10~40℃下络合4小时,得到溶液Ⅱ;
(3)将海藻酸钠、葡萄糖酸δ-内酯、碳酸钙纳米颗粒溶于步骤(1)的溶液I或步骤(2)的溶液Ⅱ中,磁力搅拌至固体粉末全部溶解,得到混合分散液,再对混合溶液进行超声波脱气泡处理,混合分散液10~40℃孵育8~16h(优选为12h),得到海藻酸钠/乳清蛋白纳米纤维双网络水凝胶或负载热敏性物质的海藻酸钠/乳清蛋白纳米纤维双网络水凝胶。
本发明通过引入刚性的纳米纤维这种高长径比的增强体,利用分子之间的氢键作用形成牢固的三维网络结构,使得多糖-蛋白质水凝胶在室温下通过添加Ca2+和葡萄糖酸δ-内酯形成,提供了一种无需热处理即可封装热敏保健品化合物的机会,能运用于热敏食品中。
进一步限定,步骤(1)中所述乳清蛋白的浓度为10mg/mL-60mg/mL。
进一步限定,步骤(1)中所述加热参数为:加热温度为75~85℃(优选为80℃),加热时间为8-10h,搅拌转速为100rpm-200rpm。
进一步限定,步骤(1)中调节溶液pH至6-7。
进一步限定,步骤(2)中所述的热敏性物质可以为姜黄素、维生素、益生菌等,含量为75-250μg/mL。
进一步限定,步骤(3)中所述海藻酸钠、葡萄糖酸δ-内酯、碳酸钙纳米颗粒含量分别为0.5-3wt%、7.03-140.7mM、0.05-0.2wt%,最优选为,1wt%、28.14mM、0.1wt%。
与现有技术相比,本发明具有以下有益效果:
(1)本发明选用乳清蛋白和海藻酸钠为水凝胶原料,它们具有良好的生物相容性,同时来源广泛,制备成本较低。经过简单改性可发生交联,制备方法简单便携,常温下就可以实现双网络水凝胶的交联反应,节省能源。
(2)本发明的双网络水凝胶具有优异的机械性能、稳定性以及均匀性,还可以作为热敏物质的递送体系,同时拓宽了其应用范围,提高营养物的生物利用度。
(3)本发明使用高深宽比填料的方法来提高水凝胶机械强度,在室温下制备得到海藻酸钠/乳清蛋白纳米纤维双网络水凝胶。本发明不仅制备过程简便,而且海藻酸钠/乳清蛋白纳米纤维双网络水凝胶具有较高的凝胶强度和机械性能等显著等优点,并且可以应用于热敏营养物的递送***中。
附图说明
图1为实施例1-4和对比例1-2的水凝胶外观图。
图2为实施例1-4和对比例1的角频率扫描流变曲线图。
图3为实施例1-4和对比例1的质构特性图。
图4为实施例1、实施例4和对比例1-2以及姜黄素的傅里叶红外光谱图。
图5为实施例1、实施例4和对比例1-2以及姜黄素的示差量热扫描图。
图6为实施例1、实施例4和对比例1-2以及姜黄素的X-衍射图。
具体实施方式
下面结合具体实施例对本发明作进一步阐释,本发明根据发明技术方案进行实施,给出了详细的实施方式和操作步骤,但本发明的保护范围并不限于下述的实施例。下列实施例中未注明具体条件的实验方法,通常按照常规条件。
实施例1
一种海藻酸钠-乳清蛋白纳米纤维双网络水凝胶的制备方法,具体步骤如下:
(1)取1000mg乳清蛋白溶于20mL超纯水,配制浓度为50mg/mL的乳清蛋白溶液,调节到pH为2,并置于水浴锅中,加热温度为80℃,加热时间为8h,搅拌转速为150rpm,以促进乳清蛋白纤维形成。
(2)将步骤(1)的乳清蛋白纤维溶液调节到pH为6,然后依次按照海藻酸钠1wt%、葡萄糖酸δ-内酯28.14mM、碳酸钙纳米颗粒0.1wt%加入,磁力搅拌至固体全部溶解之后进行超声脱气泡。混合分散液室温25℃下放置12h,得到双网络水凝胶。
取上述制备的双网络水凝胶,进行指标测定,结果如图1-6所示。
实施例2
本实例除步骤(1)中乳清蛋白溶液的浓度为10mg/mL,其余内容与实施例1完全相同。
实施例3
本实例除步骤(1)中乳清蛋白溶液的浓度为60mg/mL,其余内容与实施例1完全相同。
实施例4
一种负载姜黄素的海藻酸钠/乳清蛋白纳米纤维双网络水凝胶的制备方法,具体步骤如下:
(1)取1000mg乳清蛋白溶于20mL超纯水,配制浓度为50mg/mL的乳清蛋白溶液,调节到pH为2,并置于水浴锅中,加热温度为80℃,加热时间为8h,搅拌转速为150rpm,以促进乳清蛋白纤维形成。
(2)将步骤(1)的乳清蛋白纤维溶液调节到pH为6,在75%的乙醇水溶液中制备浓度为80mg/mL的姜黄素悬浮液,使最终聚合物溶液中的乙醇浓度小于0.2%(v/v)。最终姜黄素的浓度为125μg/mL,将该悬浮液滴加到乳清蛋白纳米纤维溶液中。混合物于室温下在磁力搅拌器上保存4小时。
(3)搅拌好的混合分散液依次按照海藻酸钠1wt%、葡萄糖酸δ-内酯28.14mM、碳酸钙纳米颗粒0.1wt%加入,磁力搅拌至固体全部溶解之后进行超声脱气泡。混合分散液室温下放置12h,得到负载姜黄素的双网络水凝胶。
之后进行指标构测定,结果如图1~6所示。
对比例1
一种分子交联海藻酸钠水凝胶的制备方法,具体步骤如下:
(1)取20mL超纯水,将pH调到6。
(2)然后依次按照海藻酸钠1wt%、葡萄糖酸δ-内酯28.14mM、碳酸钙纳米颗粒0.1wt%加入,磁力搅拌至固体全部溶解之后进行超声脱气泡。混合分散液室温下放置12h,得到交联海藻酸钠分子水凝胶。
进行指标测定,结果如图1-6所示。
对比例2
一种乳清蛋白纳米纤维的制备方法,具体步骤如下:
取1000mg乳清蛋白溶于20mL超纯水,配制浓度为50mg/mL的乳清蛋白溶液,调节到pH为2,并置于水浴锅中,加热温度为80℃,加热时间为8h,搅拌转速为150rpm,以促进乳清蛋白纤维形成。
如图1A所示,实施例1-4形成的水凝胶具有固体的外部形状,对比例1形成的水凝胶成透明状,对比例2仍为透明液体。图1B可知,采用旋转倒置法(>10min,稳态)实施例1-4及对比例1可以阻止凝胶沿引力方向流动,反应牢固,而对比例2表现为低粘性液体,没有显示出凝胶的迹象。
由图2可知,实施例1-4及对比例1均产生了类固体的行为,储能模量均大于损耗模量,都是弹性凝胶。实施例1、实施例4储能模量和损耗模量均高于实施例2和实施例3,表明乳清蛋白纤维浓度在50mg/mL时水凝胶增强效果最好。同时,实施例1的储能模量约是对比例1的6倍,这表明海藻酸钠/乳清蛋白纳米纤维双网络水凝胶比单网络结构水凝胶具有更强的凝胶强度。
由图3可知,实施例1的TPA性能较实施例2、实施例3以及实施例4都好,实施例1中海藻酸钠和乳清蛋白纳米纤维变为双连续模式,两者形成了强而内聚的双网络,TPA性能强。实施例2中凝胶网络结构主要由海藻酸钠网络确定,导致TPA相对较差。实施例3形成的水凝胶变成了乳清蛋白纳米纤维单连续模式,TPA下降。实施例4说明姜黄素的存在使蛋白质结构变松散从而使TPA降低。实施例1与对比例1比较,可以得出双网络水凝胶的TPA性能可以显著增强。
如图4所示,,通过傅里叶红外光谱图发现,实施例4中姜黄素的吸收峰被覆盖,说明姜黄素被包埋于水凝胶内部。在对比例1-2和姜黄素的光谱中,羟基对应的位置分别在3418cm-1,3280cm-1和3500cm-1处,在实施例4中消失,说明姜黄素可能通过氢键或疏水作用与乳清蛋白纳米纤维和海藻酸钠相互作用。对比例2中在1660cm-1和1530cm-1处是酰胺Ⅰ键和酰胺Ⅱ键,加入海藻酸钠后,在实施例1中观察到峰的位置变成了1607cm-1,1405cm-1表明两者之间存在静电相互作用和氢键,而在加入姜黄素后实施例4中峰的位置变成了1633cm-1和1498cm-1,表明了三者之间的静电相互作用。
如图5所示,实施例1、实施例4以及对比例1-2在30到150℃加热过程中都出现广泛吸热峰特征,对比例2在69.08℃出现了内热峰,可能因为β-乳球蛋白变性。实施例1的内热峰变为71.08℃,变性温度升高可能海藻酸钠增强了与乳清蛋白之间的疏水效应和静电相互作用。加入了姜黄素,实施例4中变性温度略降低至69.5℃,这因为姜黄素的结合使蛋白质结果变得松散,从而使变性温度降低。
如图6所示,通过X射线衍射图发现,实施例1、实施例4以及对比例1在2θ=13.5°,31.6°,45.3°处有三个相似的衍射峰。在18.1°,8.8°,17.3°,12.1°,21.1°,25.5°,23.4°等处的峰属于姜黄素的晶型衍射峰,表面姜黄素以晶型状态存在。姜黄素的晶型衍射峰在实施例4中消失,表明姜黄素在水凝胶中变为无定形状。
Claims (5)
1.一种负载功能成分双网络水凝胶的制备方法,其特征在于,该制备方法按以下步骤进行:
步骤1:将乳清蛋白溶于超纯水中,在pH为1.5~2.5的条件下加热搅拌,调节溶液pH至6-7,得到溶液I;
所述加热搅拌的条件为:加热温度为75~85℃,加热时间为8-10h,搅拌转速为100rpm-200rpm;
步骤2:滴加热敏性物质到步骤1的溶液I中,将混合物放置络合2~6小时,得到溶液Ⅱ;
步骤3:将海藻酸钠、葡萄糖酸δ-内酯、碳酸钙纳米颗粒溶于步骤1的溶液I中或步骤2的溶液Ⅱ中,磁力搅拌至固体粉末全部溶解,得到混合分散液,再对混合溶液进行超声波脱气泡处理,混合分散液10~40℃孵育8~16h,得到海藻酸钠/乳清蛋白纳米纤维双网络水凝胶或者负载热敏性物质的海藻酸钠/乳清蛋白纳米纤维双网络水凝胶,即负载功能成分双网络水凝胶。
2.如权利要求1所述的负载功能成分双网络水凝胶的制备方法,其特征在于,步骤1中,所述溶液I的乳清蛋白的浓度为10mg/mL-60mg/mL。
3.如权利要求1所述的负载功能成分双网络水凝胶的制备方法,其特征在于,步骤2中,所述的热敏性物质为姜黄素、维生素或益生菌。
4.如权利要求1所述的负载功能成分双网络水凝胶的制备方法,其特征在于,步骤2中,所述的热敏性物质在溶液Ⅱ中的含量为75-250μg/mL。
5.如权利要求1所述的负载功能成分双网络水凝胶的制备方法,其特征在于,步骤3中,所述海藻酸钠、葡萄糖酸δ-内酯、碳酸钙纳米颗粒在溶液I或溶液Ⅱ中的含量分别为0.5-3.0wt%、7.03-140.7mM、0.05-0.2wt%。
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