CN115336760B - 一种基于交叉水凝胶构建的超稳定混合型高内相乳液体系及其制备方法 - Google Patents
一种基于交叉水凝胶构建的超稳定混合型高内相乳液体系及其制备方法 Download PDFInfo
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Abstract
本发明属于生物制品加工技术领域,具体涉及一种基于交叉水凝胶构建的超稳定混合型高内相乳液体系及其制备方法,包括连续相和分散相;连续相是由三元复合物1和三元复合物2通过自组装构建的交叉水凝胶;三元复合物1由功能蛋白、亲水性多酚、阴离子多糖自组装而成;三元复合物2由功能蛋白、疏水性多酚、阴离子多糖自组装而成;分散相为2种油脂,油脂1为功能性油脂,其位于三元复合物1内;油脂2为负载功能性色素的油脂,其位于三元复合物2内。该方法制备工艺简单,混合型高内相乳液具有极其优异的稳定性,可以作为新型功能性配料或超稳定运载体系,用于共包埋、共传递和共运载各种类型功能因子,可广泛应用于食品、药品和化妆品中。
Description
技术领域
本发明属于生物制品加工技术领域,具体涉及一种基于交叉水凝胶构建的超稳定混合型高内相乳液体系及其制备方法。
背景技术
近年来,超稳定乳液体系因具有更优异的稳定性(如抗环境应力、聚结等)和对生物活性化合物具有更好保护作用,受到了研究人员的广泛关注。与其它乳液体系相比,高内相乳液(HIPEs)是构建超稳定乳液体系的理想平台。当内相体积分数超过0.74时,分散液滴达到其最大堆积密度,即可形成具有高粘弹性的HIPEs。到目前为止,各种食品级和环境友好的生物基复合物(如蛋白质-多糖复合物、蛋白质-多酚复合物和多糖-多酚复合物)已被用于稳定HIPEs。然而,这些基于生物基复合物的HIPEs在动力学或热力学上仍然不稳定。
提高分散和/或连续相网络的交联度被认为是进一步提高HIPEs稳定性的有效策略。一些研究通过在分散相和连续相中添加高粘度的甘油单油酸酯、蜂蜡等进行网络化,另一些研究通过谷氨酰胺转胺酶和钙离子交联醇溶蛋白-海藻酸钠凝聚粒子稳定乳液。但这些研究操作繁琐,且所构建的乳液体系仍旧会在2个月内失稳,稳定性仍有待提高。
通过将两种带相反电荷的乳液简单混合,在某些条件下会产生协同效应。例如,在5<pH<8条件下,β-乳球蛋白基乳液和乳铁蛋白基乳液能够通过静电吸引形成异质聚集乳液,在一定程度上改善了乳液体系的稳定性和质地特性。而由酪蛋白酸钠基乳液和乳铁蛋白基乳液混合形成的异质聚集乳液,其脂质消化程度低,可用于控制能量摄入。然而,这些异质聚集乳液的连续相组成往往较为单一,乳液体系结构不易调整;所运载的功能因子类型较为单一;更重要的是,这些异聚行为都只进行了短期储存(1天或12小时)评估,所构建出的混合乳液并不稳定,难以满足对超稳定乳液体系的需求。
发明内容
为了解决上述技术问题,本发明提供了一种基于交叉水凝胶构建的超稳定混合型HIPEs体系及其制备方法。
本发明具体是通过如下技术方案来实现的。
本发明的第一个目的是提供一种基于交叉水凝胶构建的超稳定混合型HIPEs体系,包括连续相和分散相;
所述连续相是由三元复合物1和三元复合物2通过自组装构建的交叉水凝胶;所述三元复合物1由功能蛋白、亲水性多酚、阴离子多糖自组装而成;所述三元复合物2由功能蛋白、疏水性多酚、阴离子多糖自组装而成;
所述分散相为2种油脂,油脂1为功能性油脂,油脂1位于所述三元复合物1内;油脂2为负载功能性色素的油脂,油脂2位于所述三元复合物2内。
优选的,所述阴离子多糖为κ-卡拉胶、低甲酯果胶、海藻酸钠中的一种或几种。
优选的,所述功能蛋白为乳铁蛋白,所述亲水性多酚为表没食子儿茶素没食子酸酯(EGCG),所述疏水性多酚为姜黄素。
更优选的,所述三元复合物1由乳铁蛋白、EGCG、κ-卡拉胶自组装而成;所述三元复合物2由乳铁蛋白、姜黄素、低甲酯果胶自组装而成。
优选的,所述油脂1为牡丹籽油,油脂2为含有β-胡萝卜素的中链甘油三酯(MCT)。
本发明第二个目的是提供上述基于交叉水凝胶构建的超稳定混合型HIPEs体系的制备方法,包括以下步骤:
S1、以功能蛋白水溶液、亲水性多酚水溶液和阴离子多糖水溶液为原料,先将其中两种原料混合自组装,形成的复合物再与第三种原料混合自组装,形成三元复合物1;
以功能蛋白水溶液、疏水性多酚乙醇溶液和阴离子多糖水溶液为原料,先将其中两种原料混合自组装,形成的复合物再与第三种原料混合自组装,形成三元复合物2;
S2、将S1三元复合物1与油脂1混合,并在5000-12000rpm的剪切速率下剪切,制备高内相乳液1;
将S1三元复合物2与油脂2混合,并在5000-12000rpm的剪切速率下剪切,制备高内相乳液2;
S3、将S2中高内相乳液1和高内相乳液2搅拌混合,制得超稳定混合型HIPEs体系。
优选的,S1中的三元复合物1和三元复合物2,功能蛋白的质量分数分别为0.1%-5%,功能蛋白与阴离子多糖的质量比分别为0.1-50:1,功能蛋白与亲水性多酚或疏水性多酚的质量比分别为50-200:1。
优选的,S1中,自组装的方法是先将两种原料涡旋振荡,静置组装0.5-20h后,再与第三种原料涡旋振荡,再静置组装0.5-20h。
优选的,S2中,在制备高内相乳液1和高内相乳液2时,剪切时间分别为1-20min;高内相乳液1和高内相乳液2中的油水体积比分别为(95:5)-(74:26);
S3中,高内相乳液1和高内相乳液2的体积比为(1:9)-(9:1),搅拌时间为1-30min。
本发明的第三个目的是提供基于交叉水凝胶构建的超稳定混合型HIPEs体系在食品、药品和化妆品中的应用。
本发明与现有技术相比具有如下有益效果:
本发明首先制备了两种带负电荷的HIPEs:分散相分别为2种油脂(功能性油脂和负载功能性色素的油脂),连续相分别为三元复合物1和三元复合物2;三元复合物1由功能蛋白、亲水性多酚、阴离子多糖自组装而成;三元复合物2由功能蛋白、疏水性多酚、阴离子多糖自组装而成。之后,将制备好的这两种带负电荷的HIPEs简单混合,即可制备得到具有极其优异稳定性的混合型HIPEs;其中,这种混合型HIPEs的连续相中存在以下协同作用:乳铁蛋白、亲水性多酚、疏水性多酚、两种阴离子多糖通过自组装形成交叉水凝胶,凭借强大的空间位阻作用、静电排斥作用和优异的粘弹特性,有效防止了液滴直接接触和聚结,极大提高了乳液的稳定性。
其中,上述包括以下两种带负电荷的HIPEs:一种是以牡丹籽油为油相,由乳铁蛋白、EGCG和κ-卡拉胶自组装形成的非共价三元复合物稳定的HIPEs;另一种是以含有β-胡萝卜素的MCT为油相,由乳铁蛋白、姜黄素和低甲酯果胶自组装形成的非共价三元复合物稳定的HIPEs,将这两种HIPEs组合,得到九种混合型HIPEs,经过检测,制备的所有混合型HIPEs都是超稳定的,并且实现了多种生物活性物质(即1种功能蛋白、1种亲水性多酚、1种疏水性多酚、1种功能性油脂和1种功能性色素)的共运载,大大提高了它们的稳定性和控释效果,为功能因子稳态化提供新途径;新型超稳定混合型HIPEs可应用于高价值功能食品、化妆品和生物医学领域。
附图说明
图1为各实施例制备的混合型HIPEs储存1天的表观对照图;
图2为各实施例制备的混合型HIPEs弹性(EI值)、粘度(MVI值)和固液平衡值(SLB值)的对照图;
图3为各实施例制备的混合型HIPEs的3D打印特性图;
图4为各实施例制备的混合型HIPEs的光学显微镜图片;
图5为各实施例制备的混合型HIPEs的长期储存(12个月)的表观对照图;
图6为混合型HIPEs稳定性的对比图;
图7为各实施例制备的混合HIPEs的连续相溶液状态及模型结构图;
图8为单独κ-卡拉胶溶液、单独低甲酯果胶溶液、κ-卡拉胶溶液+低甲酯果胶混合溶液的表观图片;
图9为各实施例制备的混合HIPEs形成机制图;
图10为各实施例制备的混合HIPEs的自由基清除能力图;
图11为各实施例制备的混合HIPEs的β-胡萝卜素光降解行为图;
图12为各实施例制备的混合HIPEs的β-胡萝卜素光降解行为表观图;
图13为各实施例制备的混合HIPEs的油脂氧化图;
图14为各实施例制备的混合HIPEs的EGCG释放特性图;
图15为各实施例制备的混合HIPEs的姜黄素释放特性图;
图16为各实施例制备的混合HIPEs的β-胡萝卜素释放特性图;
简写说明:
“κ1”指“基于乳铁蛋白-κ-卡拉胶-EGCG三元复合物稳定的HIPEs”;
“κ2”指“基于乳铁蛋白-EGCG-κ-卡拉胶三元复合物稳定的HIPEs”;
“κ3”指“基于κ-卡拉胶-EGCG-乳铁蛋白三元复合物稳定的HIPEs”;
“L1”指“基于乳铁蛋白-低甲酯果胶-姜黄素三元复合物稳定的HIPEs”;
“L2”指“基于乳铁蛋白-姜黄素-低甲酯果胶三元复合物稳定的HIPEs”;
“L3”指“基于低甲酯果胶-姜黄素-乳铁蛋白三元复合物稳定的HIPEs”;
“CS1”指“基于乳铁蛋白-壳聚糖-姜黄素三元复合物稳定的HIPEs”;
“CS2”指“基于乳铁蛋白-姜黄素-壳聚糖三元复合物稳定的HIPEs”;
“CS3”指“基于壳聚糖-姜黄素-乳铁蛋白三元复合物稳定的HIPEs”;
“OG1”指“基于乳铁蛋白-燕麦β-葡聚糖-姜黄素三元复合物稳定的HIPEs”;
“OG2”指“基于乳铁蛋白-姜黄素-燕麦β-葡聚糖三元复合物稳定的HIPEs”;
“OG3”指“基于燕麦β-葡聚糖-姜黄素-乳铁蛋白三元复合物稳定的HIPEs”;
“κ1+CS”表示:κ1与CS(CS1、CS2、CS3)的混合乳液;
“κ1+OG”表示:κ1与OG(OG1、OG2、OG3)的混合乳液;
“κ1κ2”、“κ1κ3”和“κ2κ3”分别表示:由κ1、κ2和κ3两两之间混合所形成的混合乳液;
“L1L2”、“L1L3”和“L2L3”表示:由L1、L2和L3两两之间混合所形成的混合乳液;
“κ1L1”、“κ1L2”、“κ1L3”表示:由κ1分别与L1、L2和L3混合所形成的混合乳液;
“κ2L1”、“κ2L2”、“κ2L3”表示:由κ2分别与L1、L2和L3混合所形成的混合乳液;
“κ3L1”、“κ3L2”、“κ3L3”表示:由κ3分别与L1、L2和L3混合所形成的混合乳液。
具体实施方式
为了使本领域技术人员更好地理解本发明的技术方案能予以实施,下面结合具体实施例和附图对本发明作进一步说明,但所举实施例不作为对本发明的限定。下述各实施例中所述实验方法和检测方法,如无特殊说明,均为常规方法;所述试剂和材料,如无特殊说明,均可在市场上购买得到。
实施例1
一种基于交叉水凝胶构建的超稳定混合型HIPEs体系的制备方法,包括以下步骤:
(1)溶解:
①将0.5g乳铁蛋白、1gκ-卡拉胶、0.2g EGCG分别溶解于pH 5.0的醋酸盐缓冲液中,配置成浓度为3.2%(w/v)、1.9%(w/v)、8.0mM的水溶液;缓冲溶液中含有质量浓度3.0%氯化钠,当氯化钠含量较低时(如0.0%、0.1%和0.2%时),下述步骤(2)中自组装构建的三元复合物,体系有大量沉淀,当含量较高时,制备的乳液粘弹性差;
②将0.5g乳铁蛋白、1g低甲酯果胶分别溶解于pH 5.0的醋酸盐缓冲液中,配置成浓度为3.2%(w/v)、4.0%(w/v)的水溶液,配置50mL10.8 mM的姜黄素乙醇溶液;
(2)自组装:分别取上述①中的乳铁蛋白、EGCG和κ-卡拉胶三种溶液6mL、0.4mL和7.1mL,按照先乳铁蛋白和κ-卡拉胶组装,再将乳铁蛋白-κ-卡拉胶复合物与EGCG自组装,涡旋振荡2min,静置组装2h,制备得到乳铁蛋白-κ-卡拉胶-EGCG三元复合物,简写为Tκ1;采用相同的方法,取②中原料,制备得到乳铁蛋白-低甲酯果胶-姜黄素三元复合物,简写为TL1;
(3)单一HIPEs的制备:将Tκ1与牡丹籽油按80:20(v/v)的油水比混合,在9000rpm下剪切2min,得到由Tκ1稳定的HIPEs(即κ1);
将TL1与含有质量浓度为1.0%β-胡萝卜素的MCT按照80:20(v/v)的油水比混合,在9000rpm下剪切2min,得到由TL1稳定的HIPEs(即L1);
(4)混合型HIPEs的制备:将步骤(3)中两种HIPEs,按照5:5的体积比,手动搅拌混合10min(此处搅拌需要轻柔,否则会使得制备的HIPEs直接破乳),之后静置2h,制得混合型HIPEs(即κ1L1)。
实施例2
一种基于交叉水凝胶构建的超稳定混合型HIPEs体系的制备方法,包括以下步骤:
(1)溶解:
①将0.5g乳铁蛋白、1gκ-卡拉胶、0.2g EGCG分别溶解于pH 5.0的醋酸盐缓冲液中,配置成浓度为3.2%(w/v)、1.9%(w/v)、8.0mM的水溶液;缓冲溶液中含有质量浓度3.0%氯化钠,当氯化钠含量较低时(如0.0%、0.1%和0.2%时),下述步骤(2)中自组装构建的三元复合物,体系有大量沉淀,当含量较高时,制备的乳液粘弹性差;
②将0.5g乳铁蛋白、1g低甲酯果胶分别溶解于pH 5.0的醋酸盐缓冲液中,配置成浓度为3.2%(w/v)、4.0%(w/v)的水溶液,配置50mL10.8 mM的姜黄素乙醇溶液;
(2)自组装:分别取上述①中的乳铁蛋白、EGCG和κ-卡拉胶三种溶液6mL、0.4mL和7.1mL,按照先乳铁蛋白和EGCG组装,再将乳铁蛋白-EGCG复合物与κ-卡拉胶自组装,涡旋振荡2min,静置组装2h,制备得到乳铁蛋白-EGCG-κ-卡拉胶三元复合物,简写为Tκ2;采用相同的方法,取②中原料,制备得到乳铁蛋白-姜黄素-低甲酯果胶三元复合物,简写为TL2;
(3)单一HIPEs的制备:将Tκ2与牡丹籽油按80:20(v/v)的油水比混合,在9000rpm下剪切2min,得到由Tκ2稳定的HIPEs(即κ2);
将TL2与含有质量浓度为1.0%β-胡萝卜素的MCT按照80:20(v/v)的油水比混合,在9000rpm下剪切2min,得到由TL2稳定的HIPEs(即L2);
(4)混合型HIPEs的制备:将步骤(3)中两种HIPEs,按照5:5的体积比,手动搅拌混合10min(此处搅拌需要轻柔,否则会使得制备的HIPEs直接破乳),之后静置2h,制得混合型HIPEs(即κ2L2)。
实施例3
一种基于交叉水凝胶构建的超稳定混合型HIPEs体系的制备方法,包括以下步骤:
(1)溶解:
①将0.5g乳铁蛋白、1gκ-卡拉胶、0.2g EGCG分别溶解于pH 5.0的醋酸盐缓冲液中,配置成浓度为3.2%(w/v)、1.9%(w/v)、8.0mM的水溶液;缓冲溶液中含有质量浓度3.0%氯化钠,当氯化钠含量较低时(如0.0%、0.1%和0.2%时),下述步骤(2)中自组装构建的三元复合物,体系有大量沉淀,当含量较高时,制备的乳液粘弹性差;
②将0.5g乳铁蛋白、1g低甲酯果胶分别溶解于pH 5.0的醋酸盐缓冲液中,配置成浓度为3.2%(w/v)、4.0%(w/v)的水溶液,配置50mL10.8 mM的姜黄素乙醇溶液;
(2)自组装:分别取上述①中的乳铁蛋白、EGCG和κ-卡拉胶三种溶液6mL、0.4mL和7.1mL,按照先κ-卡拉胶和EGCG组装,再将κ-卡拉胶-EGCG复合物与乳铁蛋白自组装,涡旋振荡2min,静置组装2h,制备得到κ-卡拉胶-EGCG-乳铁蛋白三元复合物,简写为Tκ3;采用相同的方法,取②中原料,制备得到低甲酯果胶-姜黄素-乳铁蛋白三元复合物,简写为TL3;
(3)单一HIPEs的制备:将Tκ3与牡丹籽油按80:20(v/v)的油水比混合,在9000rpm下剪切2min,得到由Tκ3稳定的HIPEs(即κ3);
将TL3与含有质量浓度为1.0%β-胡萝卜素的MCT按照80:20(v/v)的油水比混合,在9000rpm下剪切2min,得到由TL3稳定的HIPEs(即L3);
(4)混合型HIPEs的制备:将步骤(3)中两种HIPEs,按照5:5的体积比,手动搅拌混合10min(此处搅拌需要轻柔,否则会使得制备的HIPEs直接破乳),之后静置2h,制得混合型HIPEs(即κ3L3)。
实施例4
一种基于交叉水凝胶构建的超稳定混合型HIPEs体系的制备方法,包括以下步骤:
(1)溶解:
①将0.5g乳铁蛋白、1gκ-卡拉胶、0.2g EGCG分别溶解于pH 5.0的醋酸盐缓冲液中,配置成浓度为3.2%(w/v)、1.9%(w/v)、8.0mM的水溶液;缓冲溶液中含有质量浓度3.0%氯化钠,当氯化钠含量较低时(如0.0%、0.1%和0.2%时),下述步骤(2)中自组装构建的三元复合物,体系有大量沉淀,当含量较高时,制备的乳液粘弹性差;
②将0.5g乳铁蛋白、1g低甲酯果胶分别溶解于pH 5.0的醋酸盐缓冲液中,配置成浓度为3.2%(w/v)、4.0%(w/v)的水溶液,配置50mL10.8 mM的姜黄素乙醇溶液;
(2)自组装:分别取上述①中的乳铁蛋白、EGCG和κ-卡拉胶三种溶液6mL、0.4mL和7.1mL,按照先乳铁蛋白和κ-卡拉胶组装,再将乳铁蛋白-κ-卡拉胶复合物与EGCG自组装,涡旋振荡2min,静置组装2h,制备得到Tκ1;采用相同的方法,取②中原料,制备得到TL2;
(3)单一HIPEs的制备:将Tκ1与牡丹籽油按80:20(v/v)的油水比混合,在9000rpm下剪切2min,得到由Tκ1稳定的HIPEs(即κ1);
将TL2与含有质量浓度为1.0%β-胡萝卜素的MCT按照80:20(v/v)的油水比混合,在9000rpm下剪切2min,得到由TL2稳定的HIPEs(即L2);
(4)混合型HIPEs的制备:将步骤(3)中两种HIPEs,按照5:5的体积比,手动搅拌混合10min(此处搅拌需要轻柔,否则会使得制备的HIPEs直接破乳),之后静置2h,制得混合型HIPEs(即κ1L2)。
实施例5
一种基于交叉水凝胶构建的超稳定混合型HIPEs体系的制备方法,包括以下步骤:
(1)溶解:
①将0.5g乳铁蛋白、1gκ-卡拉胶、0.2g EGCG分别溶解于pH 5.0的醋酸盐缓冲液中,配置成浓度为3.2%(w/v)、1.9%(w/v)、8.0mM的水溶液;缓冲溶液中含有质量浓度3.0%氯化钠,当氯化钠含量较低时(如0.0%、0.1%和0.2%时),下述步骤(2)中自组装构建的三元复合物,体系有大量沉淀,当含量较高时,制备的乳液粘弹性差;
②将0.5g乳铁蛋白、1g低甲酯果胶分别溶解于pH 5.0的醋酸盐缓冲液中,配置成浓度为3.2%(w/v)、4.0%(w/v)的水溶液,配置50mL10.8 mM的姜黄素乙醇溶液;
(2)自组装:分别取上述①中的乳铁蛋白、EGCG和κ-卡拉胶三种溶液6mL、0.4mL和7.1mL,按照先乳铁蛋白和κ-卡拉胶组装,再将乳铁蛋白-κ-卡拉胶复合物与EGCG自组装,涡旋振荡2min,静置组装2h,制备得到Tκ1;采用相同的方法,取②中原料,制备得到TL3;
(3)单一HIPEs的制备:将Tκ1与牡丹籽油按80:20(v/v)的油水比混合,在9000rpm下剪切2min,得到由Tκ1稳定的HIPEs(即κ1);
将TL3与含有质量浓度为1.0%β-胡萝卜素的MCT按照80:20(v/v)的油水比混合,在9000rpm下剪切2min,得到TL3稳定的HIPEs(即L3);
(4)混合型HIPEs的制备:将步骤(3)中两种HIPEs,按照5:5的体积比,手动搅拌混合10min(此处搅拌需要轻柔,否则会使得制备的HIPEs直接破乳),之后静置2h,制得混合型HIPEs(即κ1L3)。
实施例6
一种基于交叉水凝胶构建的超稳定混合型HIPEs体系的制备方法,包括以下步骤:
(1)溶解:
①将0.5g乳铁蛋白、1gκ-卡拉胶、0.2g EGCG分别溶解于pH 5.0的醋酸盐缓冲液中,配置成浓度为3.2%(w/v)、1.9%(w/v)、8.0mM的水溶液;缓冲溶液中含有质量浓度3.0%氯化钠,当氯化钠含量较低时(如0.0%、0.1%和0.2%时),下述步骤(2)中自组装构建的三元复合物,体系有大量沉淀,当含量较高时,制备的乳液粘弹性差;
②将0.5g乳铁蛋白、1g低甲酯果胶分别溶解于pH 5.0的醋酸盐缓冲液中,配置成浓度为3.2%(w/v)、4.0%(w/v)的水溶液,配置50mL10.8 mM的姜黄素乙醇溶液;
(2)自组装:分别取上述①中的乳铁蛋白、EGCG和κ-卡拉胶三种溶液6mL、0.4mL和7.1mL,按照先乳铁蛋白和EGCG组装,再将乳铁蛋白-EGCG复合物与κ-卡拉胶自组装,涡旋振荡2min,静置组装2h,制备得到Tκ2;采用相同的方法,取②中原料,制备得到TL1;
(3)单一HIPEs的制备:将Tκ2与牡丹籽油按80:20(v/v)的油水比混合,在9000rpm下剪切2min,得到由Tκ2稳定的HIPEs(即κ2);
将TL1与含有质量浓度为1.0%β-胡萝卜素的MCT按照80:20(v/v)的油水比混合,在9000rpm下剪切2min,得到由TL1稳定的HIPEs(即L1);
(4)混合型HIPEs的制备:将步骤(3)中两种HIPEs,按照5:5的体积比,手动搅拌混合10min(此处搅拌需要轻柔,否则会使得制备的HIPEs直接破乳),之后静置2h,制得混合型HIPEs(即κ2L1)。
实施例7
一种基于交叉水凝胶构建的超稳定混合型HIPEs体系的制备方法,包括以下步骤:
(1)溶解:
①将0.5g乳铁蛋白、1gκ-卡拉胶、0.2g EGCG分别溶解于pH 5.0的醋酸盐缓冲液中,配置成浓度为3.2%(w/v)、1.9%(w/v)、8.0mM的水溶液;缓冲溶液中含有质量浓度3.0%氯化钠,当氯化钠含量较低时(如0.0%、0.1%和0.2%时),下述步骤(2)中自组装构建的三元复合物,体系有大量沉淀,当含量较高时,制备的乳液粘弹性差;
②将0.5g乳铁蛋白、1g低甲酯果胶分别溶解于pH 5.0的醋酸盐缓冲液中,配置成浓度为3.2%(w/v)、4.0%(w/v)的水溶液,配置50mL10.8 mM的姜黄素乙醇溶液;
(2)自组装:分别取上述①中的乳铁蛋白、EGCG和κ-卡拉胶三种溶液6mL、0.4mL和7.1mL,按照先乳铁蛋白和EGCG组装,再将乳铁蛋白-EGCG复合物与κ-卡拉胶自组装,涡旋振荡2min,静置组装2h,制备得到Tκ2;采用相同的方法,取②中原料,制备得到TL3;
(3)单一HIPEs的制备:将Tκ2与牡丹籽油按80:20(v/v)的油水比混合,在9000rpm下剪切2min,得到由Tκ2稳定的HIPEs(即κ2);
将TL3与含有质量浓度为1.0%β-胡萝卜素的MCT按照80:20(v/v)的油水比混合,在9000rpm下剪切2min,得到由TL3稳定的HIPEs(即L3);
(4)混合型HIPEs的制备:将步骤(3)中两种HIPEs,按照5:5的体积比,手动搅拌混合10min(此处搅拌需要轻柔,否则会使得制备的HIPEs直接破乳),之后静置2h,制得混合型HIPEs(即κ2L3)。
实施例8
一种基于交叉水凝胶构建的超稳定混合型HIPEs体系的制备方法,包括以下步骤:
(1)溶解:
①将0.5g乳铁蛋白、1gκ-卡拉胶、0.2g EGCG分别溶解于pH 5.0的醋酸盐缓冲液中,配置成浓度为3.2%(w/v)、1.9%(w/v)、8.0mM的水溶液;缓冲溶液中含有质量浓度3.0%氯化钠,当氯化钠含量较低时(如0.0%、0.1%和0.2%时),下述步骤(2)中自组装构建的三元复合物,体系有大量沉淀,当含量较高时,制备的乳液粘弹性差;
②将0.5g乳铁蛋白、1g低甲酯果胶分别溶解于pH 5.0的醋酸盐缓冲液中,配置成浓度为3.2%(w/v)、4.0%(w/v)的水溶液,配置50mL10.8 mM的姜黄素乙醇溶液;
(2)自组装:分别取上述①中的乳铁蛋白、EGCG和κ-卡拉胶三种溶液6mL、0.4mL和7.1mL,按照先κ-卡拉胶和EGCG组装,再将κ-卡拉胶-EGCG复合物与乳铁蛋白自组装,涡旋振荡2min,静置组装2h,制备得到Tκ3;采用相同的方法,取②中原料,制备得到TL1;
(3)单一HIPEs的制备:将Tκ3与牡丹籽油按80:20(v/v)的油水比混合,在9000rpm下剪切2min,得到由Tκ3稳定的HIPEs(即κ3);
将TL1与含有质量浓度为1.0%β-胡萝卜素的MCT按照80:20(v/v)的油水比混合,在9000rpm下剪切2min,得到由TL1稳定的HIPEs(即L1);
(4)混合型HIPEs的制备:将步骤(3)中两种HIPEs,按照5:5的体积比,手动搅拌混合10min(此处搅拌需要轻柔,否则会使得制备的HIPEs直接破乳),之后静置2h,制得混合型HIPEs(即κ3L1)。
实施例9
一种基于交叉水凝胶构建的超稳定混合型HIPEs体系的制备方法,包括以下步骤:
(1)溶解:
①将0.5g乳铁蛋白、1gκ-卡拉胶、0.2g EGCG分别溶解于pH 5.0的醋酸盐缓冲液中,配置成浓度为3.2%(w/v)、1.9%(w/v)、8.0mM的水溶液;缓冲溶液中含有质量浓度3.0%氯化钠,当氯化钠含量较低时(如0.0%、0.1%和0.2%时),下述步骤(2)中自组装构建的三元复合物,体系有大量沉淀,当含量较高时,制备的乳液粘弹性差;
②将0.5g乳铁蛋白、1g低甲酯果胶分别溶解于pH 5.0的醋酸盐缓冲液中,配置成浓度为3.2%(w/v)、4.0%(w/v)的水溶液,配置50mL10.8 mM的姜黄素乙醇溶液;
(2)自组装:分别取上述①中的乳铁蛋白、EGCG和κ-卡拉胶三种溶液6mL、0.4mL和7.1mL,按照先κ-卡拉胶和EGCG组装,再将κ-卡拉胶-EGCG复合物与乳铁蛋白自组装,涡旋振荡2min,静置组装2h,制备得到Tκ3;采用相同的方法,取②中原料,制备得到TL2;
(3)单一HIPEs的制备:将Tκ3与牡丹籽油按80:20(v/v)的油水比混合,在9000rpm下剪切2min,得到由Tκ3稳定的HIPEs(即κ3);
将TL2与含有质量浓度为1.0%β-胡萝卜素的MCT按照80:20(v/v)的油水比混合,在9000rpm下剪切2min,得到由TL2稳定的HIPEs(即L2);
(4)混合型HIPEs的制备:将步骤(3)中两种HIPEs,按照5:5的体积比,手动搅拌混合10min(此处搅拌需要轻柔,否则会使得制备的HIPEs直接破乳),之后静置2h,制得混合型HIPEs(即κ3L2)。
实施例10
步骤同实施例9,不同之处在于:
步骤(2)中,制备的Tκ3或TL2,乳铁蛋白的质量分数分别为0.1%,乳铁蛋白和低甲酯果胶或κ-卡拉胶的质量比为50:1,乳铁蛋白和姜黄素或EGCG的质量比为120:1,且静置组装时间分别为0.5h;
步骤(3)中,制备的κ3或L2,乳液的油水比分别为95:5(v/v);
步骤(4)中,两种HIPEs的体积比为1:9。
该实施例性能与实施例9近似。
实施例11
步骤同实施例9,不同之处在于:
步骤(2)中,制备的Tκ3或TL2,乳铁蛋白的质量分数分别为5%,乳铁蛋白和低甲酯果胶或κ-卡拉胶的质量比为1:10,乳铁蛋白和姜黄素或EGCG的质量比为120:1,且静置组装时间分别为20h;
步骤(3)中,制备的κ3或L2,乳液的油水比分别为74:26(v/v);
步骤(4)中,两种HIPEs的体积比为9:1。
该实施例性能与实施例9近似。
对上述实施例制备的混合型HIPEs体系进行性能表征:
图1为各实施例制备的混合型HIPEs储存1天的表观图显示:与单一HIPEs(κ1、κ2、κ3、L1、L2、L3)相比,混合型HIPEs(κ1L1、κ1L2、κ1L3、κ2L1、κ2L2、κ2L3、κ3L1、κ3L2、κ3L3)为类固体乳液。
图2为各实施例制备的混合型HIPEs的EI、MVI、SLB,显示:单一HIPEs的弹性为0.02~0.07nm-2,粘性为0.18~0.21nm-2s,SLB值为0.56~0.64(说明乳液倾向于流体特性);混合型乳液的弹性为0.11~0.32nm-2(是单一HIPEs弹性的1.9~11.5倍),粘性为0.33~0.71nm-2s(是单一HIPEs粘性的1.8~5.1倍),SLB值为0.31~0.48(说明乳液倾向于固体特性);说明将所选用的两种单一HIPEs混合,极大提高了乳液体系的粘弹特性,引起了乳液体系从流体行为到固体行为的转变;同时,通过改变蛋白质、多酚、多糖之间的结合顺序,可对乳液的弹性、粘性、SLB值进行调控。
图3为各实施例制备的混合型HIPEs的3D打印特性图;经3D打印得到的立方体表面光滑,形状完整精致,稳定性较强,说明混合型HIPEs具有较强的凝胶网络和机械强度,适合3D打印。
图4为各实施例制备的混合型HIPEs的光学显微镜图显示:混合型乳液中大小颗粒不均一混合,且大颗粒粒径明显减少。
图5为各实施例制备的混合型HIPEs的长期储存稳定性:在空气中储存12个月后,仍旧很稳定,没有油脂析出,并保持了原有颜色;而单一HIPEs,在储存1个月时,就开始有油脂析出,3个月时,油脂析出量更多,稳定性差。
图6为混合型HIPEs稳定性的对比图,由图6可见,只有当两种单一HIPEs均带相同负电荷(即,由阴离子多糖参与稳定)时,所得到的混合型HIPEs才具有超稳定性;
原因在于:电中性复合物可以吸附到油水界面,但所形成的界面层脆弱。当κ1与OG1/OG2/OG3(ζ-电位为0.8±0.2-1.2±0.4mV)混合时,形成的混合HIPEs在储存1天后严重析油,稳定性很差。一种可能的解释是,在κ1与OG1/OG2/OG3所形成的混合型HIPEs体系中,液滴之间会受到严重的挤压和堆叠,导致OG1/OG2/OG3脆弱的界面层破裂,因此使所形成的混合HIPEs不稳定。当κ1与CS1/CS2/CS3(ζ-电位为17.2±0.8mV-22.3±1.2mV)混合时,形成的混合HIPEs在储存1天后析油,表现出较低的稳定性。这与经典理论一致。即,当两个由带相反电荷壁材稳定的液滴接近时,它们相互吸引,导致壁材从油滴界面解吸,进而加速油滴间的聚结和失稳。κ1/κ2/κ3液滴和L1/L2/L3液滴具有高负电荷(均大于―20mV),因此它们之间存在强大的斥力,可以有效防止液滴直接接触和聚结。因此,液滴之间的排斥作用对混合型HIPEs的长期储存稳定性起着重要作用,不容忽视。
图7为各实施例制备的混合型HIPEs连续相溶液状态及模型结构图;连续相中形成了复杂的交叉水凝胶,交叉水凝胶的结构和功能特性(如粘弹特性)可以通过Tκ和TL的灵活组装而调控。
图8中,左图为单独κ-卡拉胶溶液,中图为单独的低甲酯果胶溶液,右图为κ-卡拉胶溶液+低甲酯果胶溶液的混合溶液,可以看出,在不添加乳铁蛋白、多酚(EGCG、姜黄素)的情况下,κ-卡拉胶溶液和低甲酯果胶溶液混合后,不能形成水凝胶,即单独这两种阴离子多糖之间不存在协同作用,需要在乳铁蛋白和EGCG、姜黄素存在的情况才能实现协同。
图9为各实施例制备的混合型HIPEs的构建机制图显示:混合型HIPEs具有超稳定性,主要是因为在该乳液的连续相中形成了由乳铁蛋白、EGCG、姜黄素、κ-卡拉胶和低甲酯果胶共同自组装构建的交叉水凝胶。
图10为各实施例混合型HIPEs的自由基清除能力显示:混合型HIPEs的自由基清除能力较高,这是由在该乳液体系中,各种功能因子(如乳铁蛋白、EGCG、姜黄素、β-胡萝卜素和牡丹籽油)之间显著的协同作用实现的。
图11为各实施例制备的混合型HIPEs中β-胡萝卜素的光降解特性显示:与单一HIPEs相比,经光照后,混合型HIPEs中的β-胡萝卜素光降解速率明显减缓,β-胡萝卜素得到了更好保护。
图12为各实施例制备的混合型HIPEs中β-胡萝卜素的光降解行为表观图。
图13为各实施例制备的混合型HIPEs的油脂氧化图;由图中数据可以看出,与单一HIPEs相比,混合型HIPEs中的油脂得到了更好保护。
图14为各实施例制备的混合型HIPEs中EGCG的释放特性图;EGCG在酸性条件下稳定,在中性条件下易于释放。
图15为各实施例制备的混合型HIPEs中姜黄素的释放特性图;姜黄素在酸性条件下稳定,在中性条件下易于释放。
图16中各实施例制备的混合型HIPEs中β-胡萝卜素缓释特性显示:在混合型HIPEs中,β-胡萝卜素在酸性条件下稳定(不易释放),在中性条件下易释放,有利于提高其生物利用率。
显然,本领域的技术人员可以对本发明进行各种改动和变型而不脱离本发明的精神和范围。这样,倘若本发明的这些修改和变型属于本发明权利要求及其等同技术的范围之内也意图包含这些改动和变型在内。
Claims (6)
1.一种基于交叉水凝胶构建的超稳定混合型高内相乳液体系,其特征在于,包括连续相和分散相;
所述连续相是由三元复合物1和三元复合物2通过自组装构建的交叉水凝胶;所述三元复合物1由功能蛋白、亲水性多酚、阴离子多糖自组装而成;所述三元复合物2由功能蛋白、疏水性多酚、阴离子多糖自组装而成;
所述分散相为2种油脂,油脂1为功能性油脂,油脂1位于所述三元复合物1内;油脂2为负载功能性色素的油脂,油脂2位于所述三元复合物2内;
所述三元复合物1中的功能蛋白为乳铁蛋白,所述亲水性多酚为表没食子儿茶素没食子酸酯,所述三元复合物1中的阴离子多糖为κ-卡拉胶;
所述三元复合物2中的功能蛋白为乳铁蛋白,所述疏水性多酚为姜黄素,所述三元复合物2中的阴离子多糖为低甲酯果胶;
所述功能性油脂为牡丹籽油,所述负载功能性色素的油脂为含有β-胡萝卜素的中链甘油三酯。
2.根据权利要求1所述的基于交叉水凝胶构建的超稳定混合型高内相乳液体系的制备方法,其特征在于,包括以下步骤:
S1、以功能蛋白水溶液、亲水性多酚水溶液和阴离子多糖水溶液为原料,先将其中两种原料混合自组装,形成的复合物再与第三种原料混合自组装,形成三元复合物1;
以功能蛋白水溶液、疏水性多酚乙醇溶液和阴离子多糖水溶液为原料,先将其中两种原料混合自组装,形成的复合物再与第三种原料混合自组装,形成三元复合物2;
S2、将S1三元复合物1与油脂1混合,并在5000-12000 rpm的剪切速率下剪切,制备高内相乳液1;
将S1三元复合物2与油脂2混合,并在5000-12000 rpm的剪切速率下剪切,制备高内相乳液2;
S3、将S2中高内相乳液1和高内相乳液2搅拌混合,制得超稳定混合型高内相乳液体系。
3.根据权利要求2所述的制备方法,其特征在于,S1中的三元复合物1和三元复合物2,功能蛋白的质量分数分别为0.1%-5%,功能蛋白与阴离子多糖的质量比分别为0.1-50:1,功能蛋白与亲水性多酚或疏水性多酚的质量比分别为50-200:1。
4.根据权利要求2所述的制备方法,其特征在于,S1中,自组装的方法是先将两种原料涡旋振荡,静置组装0.5-20 h后,再与第三种原料涡旋振荡,再静置组装0.5-20 h。
5.根据权利要求2所述的制备方法,其特征在于,S2中,在制备高内相乳液1和高内相乳液2时,剪切时间分别为1-20 min;
高内相乳液1和高内相乳液2中的油水体积比分别为(95:5)-(74:26)。
6.根据权利要求2所述的制备方法,其特征在于,S3中,高内相乳液1和高内相乳液2的体积比为(1:9)-(9:1),搅拌时间为1-30 min。
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