CN111481580A - 一种槭属多酚稳态递送纳米运载体系的制备方法 - Google Patents
一种槭属多酚稳态递送纳米运载体系的制备方法 Download PDFInfo
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Abstract
本发明提供了一种槭属多酚稳态递送纳米运载体系的制备方法。该技术方案以鱼明胶、玉米醇溶蛋白和酪蛋白酸钠为载体，采用乙醇注入和微射流技术对槭属多酚进行包载，得到槭属多酚稳态递送纳米运载体系。具体来看，本发明采用乙醇注入技术向玉米醇溶蛋白溶液中注入槭属多酚，然后与酪蛋白酸钠‑明胶混合溶液混合，并采用微射流技术对混合体系进行处理，最后制备得到一种新型的、稳定性高的槭属多酚稳态递送纳米运载体系。本发明所制备的槭属多酚稳态递送纳米运载体系，其包埋率高，贮存稳定性和消化稳定性强，在水溶液中具有良好的溶解性，而且具有更高的吸收利用率，可应用于功能性食品、保健品和护肤品等领域。
Description
技术领域
本发明涉及食品加工技术领域，具体涉及一种槭属多酚稳态递送纳米运载体系的制备方法。
背景技术
纳米运载体系是由蛋白、多糖、有机溶剂、表面活性剂等以适当比例自发形成的半透明或者不透明的悬浮稳定体系，可作为维生素、功能性油脂、益生菌和多酚类化合物等营养功效因子的稳态递送体系，广泛应用于食品、医药等领域。研究表明，不管是极性的营养素、活性因子和药物，还是非极性的营养素、活性因子和药物，合适的纳米运载体系均可提高其消化和贮存稳定性，以及加工应用范围。
多酚类化合物为普遍存在于植物中的一大类次级代谢产物，具有抗氧化、降血糖、降血脂、抗炎、抗肿瘤、抗老年斑和免疫调节等生物活性，但大部分多酚类化合物，如黄酮、单宁等，存在水溶性差，光、热、氧不稳定和消化稳定性差的缺点，极大限制了其在功能性食品、保健品和化妆品等领域的应用。因此，多酚类化合物的稳态递送运载体系的构建是当前功能性食品、化妆品等领域的研究热点。
槭属多酚为来自槭属植物茎、叶、根中的一类化合物，主要成分为黄酮、酚酸和单宁类化合物，具有抗氧化、降血糖、抗炎、抗癌等生物活性。研究发现，槭属多酚的消化稳定性非常弱，制备一种新型的槭属多酚稳态递送纳米运载体系，提高槭属多酚的消化稳定性和生物利用率，对抗氧化和降血糖等功能性产品的开发，以及槭属多酚在功能性食品和保健品中的应用具有重要意义。
发明内容
本发明旨在针对现有技术的技术缺陷，提供一种槭属多酚稳态递送纳米运载体系的制备方法，以解决现有技术中，槭属多酚贮存不稳定，光、热、氧易不稳定，消化不稳定和吸收利用率低等技术问题。
为实现以上技术目的，本发明采用以下技术方案：
一种槭属多酚稳态递送纳米运载体系的制备方法，包括以下步骤：
1)将槭属植物叶、茎或根晒干，粉碎，然后以乙醇水溶液为溶剂室温浸提三次，每次12h，得到槭属提取液；
2)将步骤1)中得到的槭属提取液合并，浓缩除去乙醇后，采用大孔吸附树脂吸附-解析，得到浓度为40％～95％(v/v)的乙醇洗脱液；
3)将步骤2)所得产物中浓度为60％～80％(v/v)的乙醇洗脱液浓缩干，得到槭属多酚组分；
4)以乙醇水溶液为溶剂溶解玉米醇溶蛋白，500rpm搅拌2h后得到玉米醇溶蛋白溶液；
5)将步骤3)所得的槭属多酚组分加入至步骤4)所得的玉米醇溶蛋白溶液中，加入过程中持续搅拌，加入后以500rpm的转速继续搅拌1h，得到槭属多酚溶液；
6)取鱼明胶和酪蛋白酸钠溶于40～50℃的水中，搅拌30min后得到鱼明胶-酪蛋白混合溶液；
7)将步骤5)所得的槭属多酚溶液逐步滴加到步骤6)所得的鱼明胶-酪蛋白混合溶液中，并采用微射流均质机处理2次进行处理，即得到所述槭属多酚稳态递送纳米运载体系。
作为优选，步骤1)中所述乙醇水溶液的浓度为60％～80％(v/v)；步骤1)中原料与溶剂的质量体积比为1:15～1:30(g:mL)。
作为优选，步骤2)中所述的吸附-解析，包括：以经过预处理的AB-8大孔吸附树脂为固定相，上样量为0.5BV，上样流速为2～3BV/h，吸附方式为静态吸附，吸附时间为8～12小时；用浓度为40％～95％(v/v)的乙醇水溶液为洗脱剂进行梯度洗脱，洗脱体积为3BV，洗脱流速为2～3BV/h。
作为优选，步骤3)中所述浓缩的温度不高于70℃。
作为优选，步骤3)所述的浓缩是在真空条件或负压条件下进行的(以增进浓缩效率)。
作为优选，步骤4)中所述乙醇水溶液的浓度75％～95％(v/v)，所述玉米醇溶蛋白的浓度为20～30mg/mL。
作为优选，步骤5)中所述槭属多酚溶液的终浓度为8～12mg/mL。
作为优选，步骤6)所述鱼明胶-酪蛋白混合溶液中，鱼明胶的质量浓度为0.1％～0.35％，酪蛋白酸钠的浓度为5mg/mL。
作为优选，步骤7)中，槭属多酚溶液和鱼明胶-酪蛋白酸钠混合溶液的体积比为1:1，动态高压微射流处理压力为80～120Mpa，处理次数为2次。
本发明提供了一种槭属多酚稳态递送纳米运载体系的制备方法。该技术方案以鱼明胶、玉米醇溶蛋白和酪蛋白酸钠为载体，采用乙醇注入和微射流技术对槭属多酚进行包载，得到槭属多酚稳态递送纳米运载体系。具体来看，本发明采用乙醇注入技术向玉米醇溶蛋白溶液中注入槭属多酚，然后与酪蛋白酸钠-明胶混合溶液混合，并采用微射流技术对混合体系进行处理，最后制备得到一种新型的、稳定性高的槭属多酚稳态递送纳米运载体系。本发明所制备的槭属多酚稳态递送纳米运载体系，其包埋率高，贮存稳定性和消化稳定性强，在水溶液中具有良好的溶解性，而且具有更高的吸收利用率，可应用于功能性食品、保健品和护肤品等领域。本发明提高了槭属多酚的水溶性，贮存和消化稳定性，扩宽其在功能性食品、保健品和护肤品等领域的应用。
与现有技术相比，本发明具有如下优点：采用60％～80％乙醇水溶液为溶剂提取槭属多酚，环境污染小，提取效率高，溶剂可回收利用；采用大孔吸附树脂柱和乙醇为洗脱溶剂对槭属多酚进行富集，富集后的大孔吸附树脂柱和乙醇均可通过活化和回收再利用，成本低，污染少，操作简单；以玉米醇溶蛋白、酪蛋白酸钠和鱼明胶为载体，采用乙醇注入法加入槭属多酚制备运载体系，多酚的包埋率高；采用微射流技术处理运载体系，解决了包载颗粒粒径大，粒度分布不均匀，稳定性差等技术难题。生产出的槭属多酚稳态递送纳米运载体系的水溶性、贮存稳定性和消化稳定性强，扩大了槭属多酚和鱼明胶在功能性食品和保健品中的应用。
具体实施方式
以下将对本发明的具体实施方式进行详细描述。为了避免过多不必要的细节，在以下实施例中对属于公知的结构或功能将不进行详细描述。以下实施例中所使用的近似性语言可用于定量表述，表明在不改变基本功能的情况下可允许数量有一定的变动。除有定义外，以下实施例中所用的技术和科学术语具有与本发明所属领域技术人员普遍理解的相同含义。
实施例1
(1)将鸡爪槭叶晒干，粉碎，然后以70％乙醇水溶液(v/v)为溶剂，按照1:20的料液比与槭属粉末混合，室温浸提三次，每次12h，得到槭属提取液；
(2)将步骤(1)中得到的提取液合并，浓缩除去乙醇后，以经过预处理的AB-8大孔吸附树脂为固定相，上样量为0.5BV，上样流速为2BV/h，吸附方式为静态吸附，吸附时间为8～12小时；用40～95％乙醇为洗脱剂进行梯度洗脱，洗脱体积为3BV，洗脱流速为2BV/h，得到40％～95％的乙醇洗脱液；
(3)将步骤(2)中得到的60％和80％的乙醇洗脱液浓于65℃进行真空浓缩，得干燥的槭属多酚组分粉末；
(4)以80％(v/v)乙醇水溶液为溶剂溶解玉米醇溶蛋白，500rpm搅拌2h后得到25mg/mL玉米醇溶蛋白溶液；
(5)称取一定量步骤(3)中制备的槭属多酚，按照一定比例缓慢加入步骤(4)中制备的玉米醇溶蛋白溶液中，边加入边搅拌，并于500rpm搅拌1h，得到浓度为12mg/mL槭属多酚溶液；
(6)分别称取一定量的鱼明胶和酪蛋白酸钠溶于40℃-50℃的温水中，搅拌30min后得到鱼明胶的浓度为0.1％，酪蛋白酸钠的浓度为5mg/mL的鱼明胶-酪蛋白混合溶液；
(7)将步骤(5)制备的槭属多酚溶液缓慢加入步骤(6)制备的鱼明胶-酪蛋白混合溶液，并采用微射流技术于120Mpa，处理2次，得到槭属多酚稳态递送纳米运载体系。
本发明得到的槭属多酚稳态递送纳米运载体系的多酚包埋率为93.6％，15天后的稳定性为93.7％，消化稳定性为77.3％，远高于未包载前的消化稳定性(40.05％)。
实施例2
(1)将元宝枫叶晒干，粉碎，然后以70％乙醇水溶液(v/v)为溶剂，按照1:20的料液比与槭属粉末混合，室温浸提三次，每次12h，得到槭属提取液；
(2)将步骤(1)中得到的提取液合并，浓缩除去乙醇后，以经过预处理的AB-8大孔吸附树脂为固定相，上样量为0.5BV，上样流速为2BV/h，吸附方式为静态吸附，吸附时间为8～12小时；用40～95％乙醇为洗脱剂进行梯度洗脱，洗脱体积为3BV，洗脱流速为2BV/h，得到40％～95％的乙醇洗脱液；
(3)将步骤(2)中得到的60％和80％的乙醇洗脱液浓于60℃进行真空浓缩，得干燥的槭属多酚组分粉末；
(4)以85％(v/v)乙醇水溶液为溶剂溶解玉米醇溶蛋白，500rpm搅拌2h后得到20mg/mL玉米醇溶蛋白溶液；
(5)称取一定量步骤(3)中制备的槭属多酚，按照一定比例缓慢加入步骤(4)中制备的玉米醇溶蛋白溶液中，边加入边搅拌，并于500rpm搅拌1h，得到浓度为10mg/mL槭属多酚溶液；
(6)分别称取一定量的鱼明胶和酪蛋白酸钠溶于40℃-50℃的温水中，搅拌30min后得到鱼明胶的浓度为0.15％，酪蛋白酸钠的浓度为5mg/mL的鱼明胶-酪蛋白混合溶液；
(7)将步骤(5)制备的槭属多酚溶液缓慢加入步骤(6)制备的鱼明胶-酪蛋白混合溶液，并采用微射流技术于100Mpa，处理2次，得到槭属多酚稳态递送纳米运载体系。
本发明得到的槭属多酚稳态递送纳米运载体系的多酚包埋率为91.2％，15天后的稳定性为91.7％，消化稳定性为77.4％，远高于未包载前的消化稳定性(38.1％)。
实施例3
(1)将五角槭叶晒干，粉碎，然后以75％乙醇水溶液(v/v)为溶剂，按照1:25的料液比与槭属粉末混合，室温浸提三次，每次12h，得到槭属提取液；
(2)将步骤(1)中得到的提取液合并，浓缩除去乙醇后，以经过预处理的AB-8大孔吸附树脂为固定相，上样量为0.5BV，上样流速为2BV/h，吸附方式为静态吸附，吸附时间为8～12小时；用40～95％乙醇为洗脱剂进行梯度洗脱，洗脱体积为3BV，洗脱流速为2BV/h，得到40％～95％的乙醇洗脱液；
(3)将步骤(2)中得到的60％和80％的乙醇洗脱液浓于60℃进行真空浓缩，得干燥的槭属多酚组分粉末；
(4)以90％(v/v)乙醇水溶液为溶剂溶解玉米醇溶蛋白，500rpm搅拌2h后得到25mg/mL玉米醇溶蛋白溶液；
(5)称取一定量步骤(3)中制备的槭属多酚，按照一定比例缓慢加入步骤(4)中制备的玉米醇溶蛋白溶液中，边加入边搅拌，并于500rpm搅拌1h，得到浓度为10mg/mL槭属多酚溶液；
(6)分别称取一定量的鱼明胶和酪蛋白酸钠溶于40℃-50℃的温水中，搅拌30min后得到鱼明胶的浓度为0.15％，酪蛋白酸钠的浓度为5mg/mL的鱼明胶-酪蛋白混合溶液；
(7)将步骤(5)制备的槭属多酚溶液采用乙醇注入的方式缓慢加入步骤(6)制备的鱼明胶-酪蛋白混合溶液，并采用微射流技术于100Mpa，处理2次，得到槭属多酚稳态递送纳米运载体系。
本发明得到的槭属多酚稳态递送纳米运载体系的多酚包埋率为91.2％，15天后的稳定性为93.5％，消化稳定性为78.6％，远高于未包载前的消化稳定性(39.5％)。
以上对本发明的实施例进行了详细说明，但所述内容仅为本发明的较佳实施例，并不用以限制本发明。凡在本发明的申请范围内所做的任何修改、等同替换和改进等，均应包含在本发明的保护范围之内。
Claims (9)
1.一种槭属多酚稳态递送纳米运载体系的制备方法，其特征在于包括以下步骤：
1)将槭属植物叶、茎或根晒干，粉碎，然后以乙醇水溶液为溶剂室温浸提三次，每次12h，得到槭属提取液；
2)将步骤1)中得到的槭属提取液合并，浓缩除去乙醇后，采用大孔吸附树脂吸附-解析，得到浓度为40％～95％(v/v)的乙醇洗脱液；
3)将步骤2)所得产物中浓度为60％～80％(v/v)的乙醇洗脱液浓缩干，得到槭属多酚组分；
4)以乙醇水溶液为溶剂溶解玉米醇溶蛋白，500rpm搅拌2h后得到玉米醇溶蛋白溶液；
5)将步骤3)所得的槭属多酚组分加入至步骤4)所得的玉米醇溶蛋白溶液中，加入过程中持续搅拌，加入后以500rpm的转速继续搅拌1h，得到槭属多酚溶液；
6)取鱼明胶和酪蛋白酸钠溶于40～50℃的水中，搅拌30min后得到鱼明胶-酪蛋白混合溶液；
7)将步骤5)所得的槭属多酚溶液逐渐滴加入至步骤6)所得的鱼明胶-酪蛋白混合溶液中，并采用微射流均质机进行2次处理，即得到所述槭属多酚稳态递送纳米运载体系。
2.根据权利要求1所述的一种槭属多酚稳态递送纳米运载体系的制备方法，其特征在于，步骤1)中所述乙醇水溶液的浓度为60％～80％(v/v)；步骤1)中原料与溶剂的质量体积比为1:15～1:30(g:mL)。
3.根据权利要求1所述的一种槭属多酚稳态递送纳米运载体系的制备方法，其特征在于，步骤2)中所述的吸附-解析，包括：以经过预处理的AB-8大孔吸附树脂为固定相，上样量为0.5BV，上样流速为2～3BV/h，吸附方式为静态吸附，吸附时间为8～12小时；用浓度为40％～95％(v/v)的乙醇水溶液为洗脱剂进行梯度洗脱，洗脱体积为3BV，洗脱流速为2～3BV/h。
4.根据权利要求1所述的一种槭属多酚稳态递送纳米运载体系的制备方法，其特征在于，步骤3)中所述浓缩的温度不高于70℃。
5.根据权利要求4所述的一种槭属多酚稳态递送纳米运载体系的制备方法，其特征在于，步骤3)所述的浓缩是在真空条件下进行的。
6.根据权利要求1所述的一种槭属多酚稳态递送纳米运载体系的制备方法，其特征在于，步骤4)中所述乙醇水溶液的浓度75％～95％(v/v)，所述玉米醇溶蛋白的浓度为20～30mg/mL。
7.根据权利要求1所述的一种槭属多酚稳态递送纳米运载体系的制备方法，其特征在于，步骤5)中所述槭属多酚溶液的终浓度为8～12mg/mL。
8.根据权利要求1所述的一种槭属多酚稳态递送纳米运载体系的制备方法，其特征在于，步骤6)所述鱼明胶-酪蛋白混合溶液中，鱼明胶溶液的质量浓度为0.1％～0.35％，酪蛋白酸钠的浓度为5mg/mL。
9.根据权利要求1所述的一种槭属多酚稳态递送纳米运载体系的制备方法，其特征在于，步骤7)中，槭属多酚溶液和鱼明胶-酪蛋白酸钠混合溶液的体积比为1:1，动态高压微射流处理压力为80～120Mpa，处理次数为2次。
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