CN109393498A - 一种包埋姜黄素的白首乌蛋白/果胶纳米鱼油乳液的制备方法 - Google Patents
一种包埋姜黄素的白首乌蛋白/果胶纳米鱼油乳液的制备方法 Download PDFInfo
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Abstract
本发明提出了一种包埋姜黄素的白首乌蛋白/果胶纳米鱼油乳液的制备方法，属于农产品深加工技术领域。该制作方法包括：将白首乌蛋白溶解于pH6‑8的PBS缓冲液，配制成5～30mg/mL的白首乌蛋白溶液；果胶溶解于水中，调节pH为6～8，配制成5～90mg/mL的果胶溶液，将蛋白储液加入到果胶储液中，搅拌10～30min得到白首乌/果胶复合水溶液。姜黄素的加入量相当于鱼油质量体积比为0.5～3g/100mL，加热并搅拌处理得到油溶液；将复合物水溶液和油溶液混合，通过高速剪切和高压均质的方法，加热处理得到稳定的乳液。本发明提供的乳液，可用于制备保健食品，药物等，具有广阔的市场发展潜力。
Description
技术领域
本发明为一种包埋姜黄素的白首乌蛋白/果胶纳米鱼油乳液的制备方法，属于农产品深加工技术领域。
背景技术
脂溶性姜黄素可以保护细胞组织，抗肿瘤，延缓衰老，在治疗和预防各种炎症和疾病方面有着显著的治疗效果，可是作为一种有效的治疗剂的广泛应用却受到了限制，原因主要归因于其超低的生物利用率及水溶性。
水包油纳米乳液是脂溶性物质很好的包埋和输送载体，可以高效地将姜黄素负载在乳滴中，姜黄素显示出更高的稳定性和水溶性，更卓越的体内吸收、控释和靶向释放性功能。
蛋白稳定的乳液对例如pH、离子强度和温度等环境压力非常敏感，当乳液中pH值接近蛋白等电点或较高盐浓度时，蛋白吸附层的静电斥力降低，因此发生聚结合絮凝。蛋白吸附层在油-水界面可以防止液滴凝聚和稳定乳液。植物蛋白是重要的蛋白资源，营养价值高，但由于植物蛋白在其等电点附近会发生沉淀，不能有效地发挥功能作用。
天然的多糖分子由于在油-水界面的吸附力差，一般不能直接作为乳化剂，但可以作为稳定剂提高乳液的稳定性，而带有一定疏水蛋白的多糖可以直接作为乳化剂。果胶作为食品上最合适结合物的阴离子多糖之一，是由α-1，4糖苷键连接半乳糖醛酸组成的复杂多糖物质，传统蛋白质乳化剂的乳化特性会受到pH和温度等因素的制约，而果胶作为一种酸性多糖可以在酸性条件下稳定蛋白质，并可以稳定酸性乳化液及乳液包埋系统。
一般蛋白质和多糖静电复合物稳定的乳液制备可通过两种方式实现，一种方式水先将蛋白质和多糖混合，制备得到蛋白质-多糖静电复合物，再与油一起进行乳化，通过该方式制备得到的乳液又被称为“混合乳状液”，另外一种方式被称为“layer-by-layer”方式，即先将单独的蛋白质和油一起乳化制备得到乳液，然后再向乳液中加入多糖，此时多糖通过静电相互作用吸附于蛋白表面。所以也被称为“双层乳状液”。但是研究发现利用“layer-by-layer”方式制备乳液存在一个很大的问题，即在乳液制备过程中常常会出现乳滴粒子之间的絮凝，即使在蛋白表面完全被多糖覆盖的情况下也会出现。通过比较两种制备乳液的方法，通过先混合蛋白与多糖形成复合物后制备得到的乳液的稳定性要比“layer-by-layer”方式制备的乳液好。据了解，包埋姜黄素的白首乌/果胶纳米鱼油乳液没有相关产业化生产的报道，在本发明中，利用白首乌蛋白与阴离子多糖果胶的静电复合作用，制备包埋有姜黄素的纳米鱼油乳液，高压均质是用于生产纳米乳剂的最有效和高通量的粉碎系统，也可以离解蛋白质胶体在不同PH值和离子强度介质中，表现出长期稳定性。
发明内容
本发明提供了一种负载姜黄素的白首乌蛋白/果胶纳米鱼油乳液的制备方法，该发明制备方法简单，原料来源广泛易得。
具体内容如下：
步骤1，原料选择、预处理：采摘、挑选新鲜饱满，肉质致密，成熟度高，无病虫害及腐烂的优质滨海白首乌，清洗、去除杂质后磨成粉状，采用溶剂低温浸提-硫酸铵沉淀法制备白首乌蛋白：原料→磨粉→石油醚脱脂→PBS缓冲液浸提→离心→取上清液→加入60％硫酸铵→静止放置→离心→沉淀→去离子水溶解→透析→冷冻干燥。
步骤2，准确称取一定量的白首乌蛋白和果胶，将白首乌蛋白溶解于pH6-8的PBS缓冲液，配制成浓度为5～30mg/mL的白首乌蛋白溶液，搅拌均匀。
步骤3，果胶溶解于水中，调节pH为6～8，配制成5～90mg/mL的果胶溶液，将白首乌蛋白储液加入到果胶储液中，放于磁力搅拌器下搅拌10～30min，混合均匀得到混合溶液，得到白首乌/果胶复合水溶液。
步骤4，将姜黄素干粉分散于鱼油中，加热并搅拌处理，得到油溶液；其中加热温度为40～65℃，鱼油的加入量相对于混合溶液的质量体积比为5～10g/100ml；姜黄素的加入量相当于鱼油质量体积比为0.5～3g/100mL；
步骤5，将上述复合物水溶液和油溶液混合，通过高速剪切和高压均质的方法，得到稳定的负载姜黄素的水包油纳米乳液。
本发明的主要优点和积极效果如下：
1、本发明提供的负载姜黄素的白首乌蛋白/果胶复合纳米鱼油乳液，在酸性条件下稳定性好，并提高了姜黄素的含量，化学稳定性及口服生物利用度。
2、本发明提供的负载姜黄素的白首乌蛋白/果胶复合纳米鱼油乳液，可用于制备保健食品，药物等。
3、本发明在制备白首乌蛋白/果胶复合乳液的过程中除了调节pH以外，没有使用化学试剂。没有加热，所使用的原料天然、安全、成本低廉。乳液的制备过程简单、绿色、省时。
具体实施方式
下面结合具体实施案例对本发明作进一步说明。
实施例1
将白首乌蛋白溶解于pH6的PBS缓冲液，配制成浓度为5mg/mL的白首乌蛋白溶液，搅拌均匀。果胶溶解于水中，调节pH为6，配制成5mg/mL的果胶溶液，将白首乌蛋白储液加入到果胶储液中，放于磁力搅拌器下搅拌10min，混合均匀得到混合溶液，溶液中加入终浓度为0.02％的叠氮钠抑制细菌生长。将姜黄素溶于鱼油中，鱼油的加入量相对于混合溶液质量体积比为5g/100ml，加热温度40℃，姜黄素的加入量相对于鱼油质量体积比为0.5g/100ml，搅拌是指在10000rpm的速度下搅拌2min；所述的均质是指利用高压微射流纳米均质机在300bar压力下均质0.5min，均质后的乳液在50℃加热0.5h，得到包埋姜黄素的白首乌蛋白/果胶纳米鱼油乳液。
实施例2
将白首乌蛋白溶解于pH7的PBS缓冲液，配制成浓度为7.5mg/mL的白首乌蛋白溶液，搅拌均匀。果胶溶解于水中，调节pH为7，配制成18.75mg/mL的果胶溶液，将白首乌蛋白储液加入到果胶储液中，放于磁力搅拌器下搅拌15min，混合均匀得到混合溶液，溶液中加入终浓度为0.02％的叠氮钠抑制细菌生长。将姜黄素溶于鱼油中，鱼油的加入量相对于混合溶液质量体积比为7g/100ml，加热温度50℃，姜黄素的加入量相对于鱼油质量体积比为1.5g/100ml，搅拌是指在10000rpm的速度下搅拌2min；所述的均质是指利用高压微射流纳米均质机在500bar压力下均质1.5min，均质后的乳液在65℃加热0.75h，得到包埋姜黄素的白首乌蛋白/果胶纳米鱼油乳液。
实施例3
将白首乌蛋白溶解于pH8的PBS缓冲液，配制成浓度为15mg/mL的白首乌蛋白溶液，搅拌均匀。果胶溶解于水中，调节pH为8，配制成37.5mg/mL的果胶溶液，将白首乌蛋白储液加入到果胶储液中，放于磁力搅拌器下搅拌30min，混合均匀得到混合溶液，溶液中加入终浓度为0.02％的叠氮钠抑制细菌生长。将姜黄素溶于鱼油中，鱼油的加入量相对于混合溶液质量体积比为10g/100ml，加热温度60℃，姜黄素的加入量相对于鱼油质量体积比为3g/100ml，搅拌是指在10000rpm的速度下搅拌2min；所述的均质是指利用高压微射流纳米均质机在800bar压力下均质3min，均质后的乳液在80℃加热1h，得到包埋姜黄素的白首乌蛋白/果胶纳米鱼油乳液。
Claims (4)
1.一种包埋姜黄素的白首乌蛋白/果胶纳米鱼油乳液的制备方法，其特征在于，是利用白首乌蛋白和果胶复合物作为乳化剂和稳定剂所制备的负载姜黄素的水包油乳液体系；其中姜黄素的包埋率为80％-95％，油相是姜黄素干粉分散于鱼油中所得的油溶液；水相是白首乌蛋白粉溶于PBS缓冲液得到的白首乌蛋白溶液。
2.一种包埋姜黄素的白首乌蛋白/果胶纳米鱼油乳液的制备方法，其特征在于，包括以下步骤：
(1)白首乌蛋白干粉的制备：
优质滨海白首乌，清洗、去除杂质后磨成粉状，采用溶剂低温浸提-硫酸铵沉淀法制备白首乌蛋白：原料→磨粉→石油醚脱脂→PBS缓冲液浸提→离心→取上清液→加入60％硫酸铵→静止放置→离心→沉淀→去离子水溶解→透析→冷冻干燥；
(2)蛋白-多糖水相的制备：
将白首乌蛋白溶解于pH6～8的PBS缓冲液，配制成浓度为5～30mg/mL的白首乌蛋白溶液，搅拌均匀。果胶溶解于水中，调节pH为6～8，配制成5～90mg/mL的果胶溶液，将白首乌蛋白储液加入到果胶储液中，放于磁力搅拌器下搅拌10～30min，混合均匀得到混合溶液，得到白首乌/果胶复合水溶液；
(3)姜黄素油溶液的制备：
将姜黄素干粉分散于鱼油中，加热并搅拌处理，得到油溶液；其中加热温度为40～60℃，鱼油的加入量相对于混合溶液的质量体积比为5～10g/100ml；姜黄素的加入量相当于鱼油质量体积比为0.5～3g/100mL；
(4)姜黄素-白首乌蛋白-果胶复合乳液的制备：
将白首乌蛋白储液加入到果胶储液中磁力搅拌混合均匀得到混合溶液；然后将姜黄素鱼油溶液与白首乌蛋白-果胶溶液进行搅拌、均质得到乳液；将均质后的乳液在50～80℃加热0.5～1h，得到包埋姜黄素的白首乌蛋白/果胶纳米鱼油乳液。
3.根据权利要求2(4)中所述包埋姜黄素的白首乌蛋白/果胶纳米鱼油乳液的制备方法，其特征在于：所述的搅拌是指在10000rpm的速度下搅拌2min；所述的均质是指利用高压微射流纳米均质机在300～900bar压力下均质0.5～3min。
4.根据权利要求1-3项所述包埋姜黄素的白首乌蛋白/果胶纳米鱼油乳液乳液，可用于制备保健食品，药物。
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