CN115068623A - 一种可稳定羊栖菜多酚的羊栖菜多糖-多酚复合物的制备方法 - Google Patents
一种可稳定羊栖菜多酚的羊栖菜多糖-多酚复合物的制备方法 Download PDFInfo
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Abstract
本发明属于天然产物和食品加工技术领域，具体涉及一种可稳定羊栖菜多酚的羊栖菜多糖‑多酚复合物的制备方法，包括以下步骤：（1）将羊栖菜多酚提取液与羊栖菜多糖溶液混合得到混合液；（2）在步骤（1）得到的混合液中添加漆酶，反应，然后进行均质处理；（3）步骤（2）处理后的反应液通过冷冻干燥得到羊栖菜多糖‑多酚复合物。本发明通过漆酶催化部分羊栖菜多酚与羊栖菜多糖共价结合，再采用均质处理，促进大分子物质羊栖菜多糖包埋羊栖菜多酚，有效制备羊栖菜多酚‑多糖复合物，可提高羊栖菜多酚在大分子物质中的包埋率和稳定性。
Description
技术领域
本发明属于天然产物和食品加工技术领域，具体涉及一种可稳定羊栖菜多酚的羊栖菜多糖-多酚复合物的制备方法。
背景技术
羊栖菜（Sargassum fusiforme）是日本、中国、韩国和朝鲜沿海地区的海洋蔬菜和药用海藻。在我国，羊栖菜有悠久的入药史，据《本草纲目》记载，羊栖菜咸能润下，寒能泄热引水，有消瘿瘤、结核，除水肿，利小便等功效。现代药理学研究表明，羊栖菜具有抗肿瘤、降血糖、降血脂、抗氧化、抗疲劳、增强免疫能力等多种生物学活性（王百龙，孙稚颖，周凤琴.羊栖菜成分以及药理研究进展. 山东中医杂志,2015, 34(9): 722-725）。羊栖菜中的主要活性成分为多糖和褐藻多酚（董玲丽，姜翠丽，柴怡，陈聪聪，李楠. 羊栖菜生物活性成分研究综述. 现代农业科技，2018, (16): 230-233），其中，多酚是一种含有多羟基的次生代谢产物，分离提取后不稳定，易氧化，对温度、光照、pH 等较为敏感。
微胶囊化是提高多酚类物质稳定性的常用技术，CN201010111965.X公开了一种采用β-环糊精、明胶、海藻酸钠、变性淀粉等的混合物为壁材，经喷雾干燥制备红景天多酚类微胶囊的方法；CN201210437395.2公开了一种采用海藻酸钠、麦芽糊精和阿拉伯胶为壁材，经过喷雾干燥获得草莓多酚微胶囊的方法；CN201811502448.8公开了一种以乳清蛋白分离物和麦芽糊精为壁材的绿茶多酚微胶囊制备方法；CN201910540633.4公开了一种利用β-环糊精为壁材制备桑椹多酚微胶囊的方法；CN202111316260.6公开了一种以魔芋粉为壁材的石榴皮渣多酚微胶囊制备方法。所采用的麦芽糊精、阿拉伯胶、β-环糊精、乳清蛋白、明胶、海藻酸钠、变性淀粉等壁材均非多酚天然结合的大分子物质，影响活性成分的稳定性，在应用上存在着一定的不足。
发明内容
本发明的目的是为了克服现有技术存在的缺点和不足，而提供一种可稳定羊栖菜多酚的羊栖菜多糖-多酚复合物的制备方法。
本发明所采取的技术方案如下：一种可稳定羊栖菜多酚的羊栖菜多糖-多酚复合物的制备方法，包括以下步骤：
（1）将羊栖菜多酚提取液与羊栖菜多糖溶液混合得到混合液；
（2）在步骤（1）得到的混合液中添加漆酶，反应，然后进行均质处理；
（3）通过冷冻干燥处理均质处理后的反应液得到羊栖菜多糖-多酚复合物。
优选的，所述羊栖菜多酚提取液与羊栖菜多糖溶液的制备过程包括以下步骤：
a. 利用液氮处理新鲜羊栖菜制得冻样后，再经粉碎得到羊栖菜粉末；
b. 将步骤a得到的羊栖菜粉末置于15%的乙醇水溶液中，在水浴中进行超声波辅助浸提处理，离心，收集上清液，上清液加入4倍乙醇，离心，沉淀即为羊栖菜多糖，上清液浓缩得到羊栖菜多酚提取液。
优选的，步骤b中，所述羊栖菜粉末加入至15%的乙醇水中的液料比为15～35mL/g。
优选的，步骤b中，所述水浴的温度为40～60℃，超声波的功率为300～600W，浸提时间为30～120min。
优选的，步骤b中，所述离心的转速为5000～12000r/min，时间为15～25min。
优选的，步骤（1）中，羊栖菜多酚提取液与羊栖菜多糖溶液以1：5-20的比例混合。
优选的，步骤（2）中，漆酶添加量为10-100U/mL。
优选的，步骤（2）中，均质处理条件如下：30-100MPa条件下均质10-30分钟。
本发明的有益效果如下：本发明通过漆酶催化羊栖菜多酚与羊栖菜多糖共价结合，再采用均质处理，促进大分子物质羊栖菜多糖包埋羊栖菜多酚，有效制备羊栖菜多酚-多糖复合物，可提高羊栖菜多酚在大分子物质中的包埋率和稳定性。
进一步的，本发明的羊栖菜粉末是利用液氮处理新鲜羊栖菜制得冻样后，再经粉碎得到，可最大限度地保护羊栖菜多酚。
具体实施方式
为使本发明的目的、技术方案和优点更加清楚，下面对本发明作进一步地详细描述。
以下实施例中，羊栖菜多酚含量、包埋率和稳定性测定方法具体入下所示：
（1）多酚含量的测定
取0.5mL适当羊栖菜提取液于试管中，加入2.5mL福林酚试剂，摇动均匀，避光反应时间为6min。后加入2mL 7.5%的Na2CO3溶液，摇动均匀后，避光，室温下放置时间为90min后，在波长765nm条件下测定吸光度。以没食子酸为标准品制作标准曲线，根据标准曲线计算多酚含量。
（2）羊栖菜多酚包埋率的测定
准确称取0.2g羊栖菜多酚-多糖复合物，用5mL无水乙醇充分冲洗，4000r/min离心3min。再根据（1）中的方法测定获得表面多酚含量（A）。
准确称取0.2g羊栖菜多酚-多糖复合物，加水1mL，于研钵中充分研磨，再加入4mL无水乙醇，超声提取5min。再根据（1）中的方法测定获得羊栖菜多酚-多糖复合物中总多酚含量（B），从而计算羊栖菜多酚包埋率。
包埋率（%）=(B-A)/B*100%
（3）稳定性测定
称取一定质量的羊栖菜多酚样品置于培养皿中，均匀分布，在30℃、光照条件下放置5天，根据（1）中的方法测定起始以及放置5天后样品总酚的含量，计算多酚保留率。
在以下实施例中，所采取的漆酶为市场上直接购买得到的，其产品推荐工艺条件为：适宜温度范围为45℃-55℃，故在以下实施例中，均采用50℃的酶反应条件。
实施例1
新鲜羊栖菜液氮处理、粉碎得到羊栖菜粉末，取羊栖菜粉末10g，加入15%的乙醇水溶液200mL，在50℃水浴中进行超声波辅助浸提处理（500W，60min），离心，收集上清液，上清液加入4倍乙醇，离心，沉淀即为羊栖菜多糖，上清液浓缩得到羊栖菜多酚提取液（5mg/mL）；羊栖菜多酚提取液与羊栖菜多糖溶液（50mg/mL）以1：10的比例混合；添加30U/mL的漆酶50°C条件下，搅拌5小时；50MPa条件下均质10分钟；冷冻干燥制得羊栖菜多糖-多酚复合物。采用实施例1的方法，测得包埋率为83.6%；在30℃、光照条件下放置5天，多酚保留率为93.2%。
实施例2
羊栖菜多糖和羊栖菜多酚的提取同实施例2。羊栖菜多酚提取液与羊栖菜多糖溶液以1：10的比例混合；添加50U/mL的漆酶50°C条件下，搅拌5小时；30MPa条件下均质30分钟；冷冻干燥制得羊栖菜多糖-多酚复合物。采用实施例1的方法，测得包埋率为86.1%；在30℃、光照条件下放置5天，多酚保留率为94.3%。
实施例3
羊栖菜多糖和羊栖菜多酚的提取同实施例2。羊栖菜多酚提取液与羊栖菜多糖溶液以1：15的比例混合；添加50U/mL的漆酶50°C条件下，搅拌3小时；50MPa条件下均质20分钟；冷冻干燥制得羊栖菜多糖-多酚复合物。采用实施例1的方法，测得包埋率为88.5%；在30℃、光照条件下放置5天，多酚保留率为92.7%。
实施例4
羊栖菜多糖和羊栖菜多酚的提取同实施例2。羊栖菜多酚提取液与羊栖菜多糖溶液以1：15的比例混合；添加50U/mL的漆酶50°C条件下，搅拌10小时；30MPa条件下均质20分钟；冷冻干燥制得羊栖菜多糖-多酚复合物。采用实施例1的方法，测得包埋率为91.4%；在30℃、光照条件下放置5天，多酚保留率为94.9%。
实施例5
羊栖菜多糖和羊栖菜多酚的提取同实施例2。羊栖菜多酚提取液与羊栖菜多糖溶液以1：20的比例混合；添加50U/mL的漆酶50°C条件下，搅拌5小时；30MPa条件下均质30分钟；冷冻干燥制得羊栖菜多糖-多酚复合物。采用实施例1的方法，测得包埋率为91.9%；在30℃、光照条件下放置5天，多酚保留率为95.2%。
实施例6
羊栖菜多糖和羊栖菜多酚的提取同实施例2。羊栖菜多酚提取液与羊栖菜多糖溶液以1：20的比例混合；添加100U/mL的漆酶50°C条件下，搅拌3小时；50MPa条件下均质20分钟；冷冻干燥制得羊栖菜多糖-多酚复合物。采用实施例1的方法，测得包埋率为93.5%；在30℃、光照条件下放置5天，多酚保留率为95.8%。
以上所揭露的仅为本发明较佳实施例而已，当然不能以此来限定本发明之权利范围，因此依本发明权利要求所作的等同变化，仍属本发明所涵盖的范围。
Claims (8)
1.一种可稳定羊栖菜多酚的羊栖菜多糖-多酚复合物的制备方法，其特征在于包括以下步骤：
（1）将羊栖菜多酚提取液与羊栖菜多糖溶液混合得到混合液；
（2）在步骤（1）得到的混合液中添加漆酶，反应，然后进行均质处理；
（3）通过冷冻干燥处理均质处理后的反应液得到羊栖菜多糖-多酚复合物。
2.根据权利要求1所述的可稳定羊栖菜多酚的羊栖菜多糖-多酚复合物的制备方法，其特征在于：所述羊栖菜多酚提取液与羊栖菜多糖溶液的制备过程包括以下步骤：
a. 利用液氮处理新鲜羊栖菜制得冻样后，再经粉碎得到羊栖菜粉末；
b. 将步骤a得到的羊栖菜粉末置于15%的乙醇水溶液中，在水浴中进行超声波辅助浸提处理，离心，收集上清液，上清液加入4倍乙醇，离心，沉淀即为羊栖菜多糖，上清液浓缩得到羊栖菜多酚提取液。
3.根据权利要求2所述的可稳定羊栖菜多酚的羊栖菜多糖-多酚复合物的制备方法，其特征在于：步骤b中，所述羊栖菜粉末加入至15%的乙醇水中的液料比为15～35mL/g。
4.根据权利要求2所述的可稳定羊栖菜多酚的羊栖菜多糖-多酚复合物的制备方法，其特征在于：步骤b中，所述水浴的温度为40～60℃，超声波的功率为300～600W，浸提时间为30～120min。
5.根据权利要求2所述的可稳定羊栖菜多酚的羊栖菜多糖-多酚复合物的制备方法，其特征在于：步骤b中，所述离心的转速为5000～12000r/min，时间为15～25min。
6.根据权利要求2所述的可稳定羊栖菜多酚的羊栖菜多糖-多酚复合物的制备方法，其特征在于：步骤（1）中，羊栖菜多酚提取液与羊栖菜多糖溶液以1：5-20的比例混合。
7.根据权利要求1所述的可稳定羊栖菜多酚的羊栖菜多糖-多酚复合物的制备方法，其特征在于：步骤（2）中，漆酶添加量为10-100U/mL。
8.根据权利要求1所述的可稳定羊栖菜多酚的羊栖菜多糖-多酚复合物的制备方法，其特征在于：步骤（2）中，均质处理条件如下：30-100MPa条件下均质10-30分钟。
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