CN111631385A - 一种荷载姜黄素Pickering乳液及其制备方法 - Google Patents
一种荷载姜黄素Pickering乳液及其制备方法 Download PDFInfo
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Abstract
本发明公开了一种以糖基化乳清分离蛋白‑壳聚糖静电复合物作为稳定剂制备荷载姜黄素Pickering乳液的方法。该方法包括如下步骤:将乳清分离蛋白糖基化,得到糖基化乳清分离蛋白;将所述糖基化乳清分离蛋白与壳聚糖混合,制备成糖基化蛋白‑壳聚糖复合物溶液;将所述糖基化蛋白‑壳聚糖复合物溶液与姜黄素溶液混合,得到所述荷载姜黄素Pickering乳液。本发明的Pickering乳液液滴呈现出良好的均一性和储藏稳定性。糖基化乳清分离蛋白‑壳聚糖复合物在负载活性物质的油滴外部形成更致密和更厚的界面层,阻碍了乳液液滴的聚并反应,抑制了影响因子(光,热,氧)造成的活性物质的降解。
Description
技术领域
本发明属于食品科学与工程技术领域,主要涉及一种以糖基化乳清分离蛋白-壳聚糖静电复合物作为稳定剂制备荷载姜黄素Pickering乳液的方法。
背景技术
蛋白质属于高分子并且具备两亲结构,所以具有独特的表面活性,但是在食品工业上的应用受环境因素限制,因为蛋白质在遇热或存在有机溶剂的环境下等不稳定。加入多糖可有效改善蛋白功能特性,一方面,两者可通过静电相互作用形成蛋白质-多糖复合物,这类复合物的增稠性和凝胶行为可以增强胶体的稳定性;另一方面,蛋白与多糖发生美拉德反应,产物对pH、温度和盐离子浓度具有良好的稳定性。
姜黄素是来自姜黄的一种天然黄色颜料,具有抗炎,抗癌,抗糖尿病,抗衰老等生物活性。姜黄素水溶性差,并且易降解,因此在食品工业中的应用受到限制。食品科学领域中对姜黄素的乳液运输系统十分关注,但市场上的姜黄素负载乳液稳定性易受环境影响,造成乳液聚集和活性物质降解,最终丧失生物活性。因此,需要一种新的用于负载姜黄素的乳液。
发明内容
为解决以上问题,本发明以糖基化乳清分离蛋白-壳聚糖静电复合物为稳定剂制备一种姜黄素负载Pickering乳液。复合物可以在油滴外部形成致密界面层明显提高姜黄素在乳液中的降解稳定性。
为了实现上述目的,本发明提供了一种制备荷载姜黄素Pickering乳液的方法,其特征在于,包括以下步骤:
S1:将乳清分离蛋白糖基化,得到糖基化乳清分离蛋白;
S2:将所述糖基化乳清分离蛋白与壳聚糖混合,制备成糖基化蛋白-壳聚糖复合物溶液;
S3:将所述糖基化蛋白-壳聚糖复合物溶液与姜黄素溶液混合,得到所述荷载姜黄素Pickering乳液。
在一个优选实施方案中,S1中,将乳清分离蛋白与D-(+)乳糖水溶液溶于水中,得到乳清分离蛋白和D-(+)乳糖的混合溶液,然后将所述乳清分离蛋白和D-(+)乳糖的混合溶液冻干成粉末,孵化12-24h,即得到所述糖基化乳清分离蛋白。
优选地,孵化时间为24h。
在一个优选实施方案中,所述乳清分离蛋白和D-(+)乳糖的混合溶液中,乳清分离蛋白与D-(+)乳糖的总质量为5%~10%。
优选地,乳清分离蛋白与D-(+)乳糖的质量比为1:1。
在一个优选实施方案中,所述乳清分离蛋白和D-(+)乳糖的混合溶液的pH可使用例如氢氧化钠调节为7。
在一个优选实施方案中,所述乳清分离蛋白和D-(+)乳糖的混合溶液冻干成的粉末在湿度为79%、温度为70℃的环境下孵化过夜。
在一个优选实施方案中,S2中将所述糖基化乳清分离蛋白溶于水中,加入壳聚糖,混合后得到所述糖基化蛋白-壳聚糖复合物溶液。
优选地,糖基化乳清分离蛋白与壳聚糖混合过程中,可使用例如盐酸将体系的pH调节至5。
在一个优选实施方案中,所述糖基化蛋白-壳聚糖复合物溶液中,所述糖基化乳清分离蛋白的浓度为10mg/mL。
优选地,所述糖基化乳清分离蛋白粉末与壳聚糖的质量比为1:1。
在一个优选实施方案中,S3中,将姜黄素溶于疏水有机溶剂中,得到姜黄素溶液,再将所述姜黄素溶液与所述糖基化蛋白-壳聚糖复合物混合均质,得到所述荷载姜黄素Pickering乳液。
优选地,所述疏水有机溶剂可为菜籽油、花生油、大豆油等,优选为中链脂肪酸甘油酯(MCT)
优选地,所述姜黄素溶液中姜黄素的浓度为0.05%~0.1%。
优选地,所述姜黄素溶液与糖基化蛋白-壳聚糖复合物溶液的质量比为7:3。
在一个优选实施方案中,S3中在制备姜黄素溶液和荷载姜黄素Pickering乳液的过程中还分别包括离心去沉淀的步骤。
优选地,制备姜黄素溶液所用的离心转速为10000~12000rpm,离心10~20min。制备荷载姜黄素Pickering乳液所用的离心转速为7500~15000rpm,时间为1~2min。
本发明还提供了一种荷载姜黄素Pickering乳液,由上述方法制备得到。
本发明糖基化蛋白-壳聚糖静电复合物良好的乳化特性与界面性质有效抑制了油滴间的聚集现象,使乳液液滴呈现出良好的均一性。美拉德反应改善了蛋白质的稳定性,同时糖基化改变了蛋白表面电荷密度使得壳聚糖与蛋白的静电复合更为彻底并且由于壳聚糖的加入增加了乳液粘度,糖基化乳清分离蛋白-壳聚糖静电复合物在负载活性物质的油滴外部形成更致密和更厚的界面层,阻碍了乳液液滴的聚并反应,抑制了影响因子(光,热,氧)造成的活性物质的降解。
附图说明
图1示出了本发明实施例1制备得到的糖基化乳清分离蛋白-壳聚糖纳米颗粒的粒径分布图。
图2示出了本发明实施例1制备得到的Pickering乳液的粒径分布图。
图3示出了本发明实施例1制备得到的Pickering乳液激光共聚焦图。
图4示出了本发明实施例1制备得到的Pickering乳液表观粘度与剪切速率关系图。
图5示出了本发明实施例1制备得到的姜黄素负载Pickering乳液的稳定性分析图,其中,WLC-CS表示糖基化乳清分离蛋白-壳聚糖稳定的乳液中的姜黄素含量,MCT表示游离油相中的姜黄素含量。
具体实施方式
下面将更详细地描述本发明的优选实施方式。虽然以下描述了本发明的优选实施方式,然而应该理解,可以以各种形式实现本发明而不应被这里阐述的实施方式所限制。相反,提供这些实施方式是为了使本发明更加透彻和完整,并且能够将本发明的范围完整地传达给本领域的技术人员。
实施例1
本实施例提供一种以糖基化乳清分离蛋白-壳聚糖静电复合物作为稳定剂制备荷载姜黄素Pickering乳液的方法,包括如下步骤:
1)乳清分离蛋白糖基化反应产物的制备
将乳清分离蛋白与D-(+)乳糖粉末溶于水、彻底水合后将两溶液混合均匀,得到乳清分离蛋白和D-(+)乳糖的混合溶液,将混合溶液的pH调至7,冷冻干燥,得到蛋白-乳糖混合粉末;将蛋白-乳糖混合粉末置于湿度为79%、温度为70℃的环境下反应24h,得到糖基化乳清分离蛋白粉末;
其中,乳清分离蛋白与D-(+)乳糖的质量比为1:1,乳清分离蛋白和D-(+)乳糖的混合溶液中,乳清分离蛋白与D-(+)乳糖的总质量为10%
2)糖基化蛋白-壳聚糖复合物的制备
将糖基化乳清分离蛋白粉末溶于水得到糖基化乳清分离蛋白溶液,加入壳聚糖粉末,调节溶液pH至5,充分混合即得到所述糖基化蛋白-壳聚糖复合物;
其中,糖基化乳清分离蛋白的浓度为1%(w/v),糖基化乳清分离蛋白粉末与壳聚糖的质量比为1:1。
图1为糖基化乳清分离蛋白-壳聚糖复合物的粒径分布图。可以看出由糖基化乳清分离蛋白-壳聚糖静电结合制备的纳米复合物分散性良好,粒径分布均一。
3)荷载姜黄素Pickering乳液的制备
将姜黄素溶于油相中,离心(12000rpm、15min)去除不溶物得到姜黄素溶液,再将姜黄素溶液与步骤2)的糖基化蛋白-壳聚糖静电复合物溶液混合均质(12000rpm、1min),荷载姜黄素的油相和糖基化蛋白-壳聚糖复合物溶液的水相的质量比为7:3,得到所述荷载姜黄素Pickering乳液为高粘度全乳。
上述步骤中,采用磁力搅拌的方式混合样品,并保证样品充分溶解和混匀。
图2示出了本发明一个实施例中制得的Pickering乳液的粒径分布图和光学显微镜图,图3示出了Pickering乳液的激光共聚焦图。可以看出以糖基化乳清分离蛋白-壳聚糖静电复合物为稳定剂制备的Pickering乳液的结构以及粒径分布,复合物均匀地吸附在油滴表面,形成了致密的界面层,且乳液粒径分布均一,稳定性良好。
图4示出了Pickering乳液表观粘度与剪切速率关系图与Pickering乳液样品正置,倒置外观图。可以看出剪切之前的乳液粘度大,外观呈凝胶状倒置不流动,随着剪切速度的增加,乳液粘度线性降低,说明了乳液假塑性特质。
图5为姜黄素负载Pickering乳液和游离油相中活性物质姜黄素的稳定性分析图。结果表明糖基化乳清分离蛋白-壳聚糖稳定的乳液中姜黄素半衰期为156h(WLC-CS)左右相比游离油相中姜黄素半衰期102h(MCT)左右延长了54h,表明以糖基化乳清分离蛋白-壳聚糖静电复合物为稳定剂制备的姜黄素负载Pickering乳液对活性物质姜黄素有较强的保护作用,可以有效抑制其降解。
以上已经描述了本发明的各实施例,上述说明是示例性的,并非穷尽性的,并且也不限于所披露的各实施例。凡落在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
Claims (10)
1.一种制备荷载姜黄素Pickering乳液的方法,其特征在于,包括以下步骤:
S1:将乳清分离蛋白糖基化,得到糖基化乳清分离蛋白;
S2:将所述糖基化乳清分离蛋白与壳聚糖混合,制备成糖基化蛋白-壳聚糖复合物溶液;
S3:将所述糖基化蛋白-壳聚糖复合物溶液与姜黄素溶液混合,得到所述荷载姜黄素Pickering乳液。
2.根据权利要求1所述的方法,其特征在于,S1中,将乳清分离蛋白与D-(+)乳糖水溶液溶于水中,得到乳清分离蛋白和D-(+)乳糖的混合溶液,然后将所述乳清分离蛋白和D-(+)乳糖的混合溶液冻干成粉末,孵化12-24h,即得到所述糖基化乳清分离蛋白。
3.根据权利要求2所述的方法,其特征在于,所述乳清分离蛋白和D-(+)乳糖的混合溶液中,乳清分离蛋白与D-(+)乳糖的总质量为5%~10%,乳清分离蛋白与D-(+)乳糖的质量比为1:1。
4.根据权利要求2所述的方法,其特征在于,所述乳清分离蛋白和D-(+)乳糖的混合溶液的pH调节为7。
5.根据权利要求2所述的方法,其特征在于,所述乳清分离蛋白和D-(+)乳糖的混合溶液冻干成的粉末在湿度为79%、温度为70℃的环境下孵化过夜。
6.根据权利要求1所述的方法,其特征在于,S2中将所述糖基化乳清分离蛋白溶于水中,加入壳聚糖,调节pH至5,混合后得到所述糖基化蛋白-壳聚糖复合物溶液。
7.根据权利要求6所述的方法,其特征在于,所述糖基化蛋白-壳聚糖复合物溶液中,所述糖基化乳清分离蛋白的浓度为10mg/mL,所述糖基化乳清分离蛋白粉末与壳聚糖的质量比为1:1。
8.根据权利要求1所述的方法,其特征在于,S3中,将姜黄素溶于疏水有机溶剂中,得到姜黄素溶液,再将所述姜黄素溶液与所述糖基化蛋白-壳聚糖复合物混合均质,得到所述荷载姜黄素Pickering乳液。
9.根据权利要求8所述的方法,其特征在于,所述姜黄素溶液中姜黄素的浓度为0.05%~0.1%,所述姜黄素溶液与糖基化蛋白-壳聚糖复合物溶液的质量比为7:3。
10.一种荷载姜黄素Pickering乳液,其特征在于,由权利要求1-9中任意一项所述的方法制备得到。
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