CN111001013B - 一种枇杷叶提取物纳米胶囊、制备方法及应用 - Google Patents

一种枇杷叶提取物纳米胶囊、制备方法及应用 Download PDF

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CN111001013B
CN111001013B CN201911274027.9A CN201911274027A CN111001013B CN 111001013 B CN111001013 B CN 111001013B CN 201911274027 A CN201911274027 A CN 201911274027A CN 111001013 B CN111001013 B CN 111001013B
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颜庭轩
徐霞
陈艳
张会文
赵海莲
马亮
韩新亚
严菊芬
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Abstract

本发明公开了一种枇杷叶提取物纳米胶囊、制备方法及应用，将环糊精溶解于有机溶剂中得到浓度1～50mg/mL的环糊精溶液作为提取剂；在密闭环境中，取粉碎后的枇杷叶粉末，添加环糊精溶液并通入惰性气体，超声提取，固液比范围1:5～25，将溶液过滤去除滤渣得到提取液；通过恒流泵将提取液泵入超临界高压结晶釜，结晶得到枇杷叶提取物纳米微胶囊。本发明提供的制备方法简单易行，绿色环保。全程操作条件温和，提取阶段通入不活泼气体保护活性物质不被氧化，制备提取物纳米微胶囊阶段CO2气体性质稳定，避免提取物中抗氧化类物质被氧化，利用环糊精溶液作为提取剂可有效提取枇杷叶中活性成分，得到产物有机溶剂残留极低。

Description

一种枇杷叶提取物纳米胶囊、制备方法及应用
技术领域
本发明涉及一种多酚类物质提取工艺，尤其涉及的是一种枇杷叶提取物纳米胶囊、制备方法及应用。
背景技术
枇杷叶是蔷薇科枇杷属常绿小乔木枇杷的干燥叶，是福建的一种道地性药材，味苦涩，属寒性，有清肺止咳，和胃利尿，止渴的功效。可用来治疗咳痰、肺热、咳血、胃热呕哕等。研究表明，在中成药与保健食品等药品和食品的加工方面，枇杷叶有普遍的研究和应用价值。
枇杷叶中有许多化学成分，其中主要的化学成分为多酚类，目前所知枇杷叶中主要含有没食子酸、儿茶素和咖啡酸等7种类型的多酚，这些多酚成分均有一定的抗氧化、抗菌、抗肿瘤和抗病毒作用，广泛应用于医学、农药化学及香料等行业。植物多酚是一种多羟基酚类化合物，包括单宁及相关衍生物，具有清除自由基、抗菌、抗病毒、抗肿瘤等多种生理功能，在农业生产、医学保健、化工等范围都进行了基础和应用研究，多酚的多种功能已经得到了广泛的应用，随着天然活性成分的开发利用，多酚的开发利用已经成为热门话题。枇杷叶中含有多种化学成分，这些化学成分使枇杷叶表现出多种功能特性，目前国内外对枇杷叶的功能研究主要立足于三萜酸类物质，甚少涉及到利用超声辅助环糊精溶液提取枇杷叶多酚的研究。
发明内容
本发明所要解决的技术问题在于：如何精简了提取工艺及制剂工艺流程，提供了一种枇杷叶提取物纳米胶囊、制备方法及应用。
本发明是通过以下技术方案解决上述技术问题的，本发明的一种枇杷叶提取物纳米胶囊的制备方法，包括以下步骤：
(1)将环糊精溶解于有机溶剂中得到浓度1～50mg/mL的环糊精溶液作为提取剂；
(2)将粉碎后的枇杷叶粉末置于密闭容器中，添加环糊精溶液并通入惰性气体，采用固液耦合超声辅助提取枇杷叶中多酚，并利用环糊精锥形的中空圆筒立体环状空腔结构，特异性吸附溶液中多酚类物质，待吸附平衡之后，将溶液过滤去除滤渣得到提取液；
(3)通过恒流泵将提取液泵入超临界高压结晶釜，结晶得到枇杷叶提取物纳米微胶囊。
作为本发明的优选方式之一，所述步骤(1)中，环糊精选自α-CD、β-CD、γ-CD、羟丙基-β-CD和羟乙基-β-CD中的至少一种。
作为本发明的优选方式之一，所述步骤(1)中，有机溶剂选自乙醇、丙酮、二氯甲烷、二甲基亚砜中的至少一种。
作为本发明的优选方式之一，所述步骤(2)中，超声温度30～60℃、吸附的固液比范围1:5～25，超声功率300～1000W，超声时间为10～120min。
作为本发明的优选方式之一，所述步骤(2)中，惰性气体为氮气、二氧化碳或者氩气。
作为本发明的优选方式之一，所述步骤(2)中，将CO2通入超临界高压结晶釜，同时加热超临界高压结晶釜，压力范围7～20Mpa,温度范围35～60℃，达到超临界状态并稳定至少30min后，将提取液以1～20LPM流速泵入超临界高压结晶釜，同时从进口通入CO2，从出口排出含有机溶剂的CO2，待提取液通入完毕，继续通入CO2至少3h排干有机溶剂，卸压，打开釜盖枇杷叶提取物纳米胶囊均匀覆盖于釜壁及釜底。
一种使用所述的制备方法制得的枇杷叶提取物纳米胶囊。
所述胶囊的粒径范围100～600nm，多酚含量范围为0.1～3％。
一种所述的枇杷叶提取物纳米胶囊在制备抗氧化活性的食品或药品中的应用。
环糊精的结构如下，其具有中空圆筒立体环状空腔结构，特异性吸附溶液中多酚类物质。
Figure BDA0002315036800000021
环糊精包括天然环糊精及其衍生物种类繁多常见的有α-CD、β-CD、γ-CD、羟丙基-β-CD和羟乙基-β-CD等，环糊精具有环状结构，其空洞结构中可有效包裹药物小分子，而其外部含有大量亲水基团因此既可以有效包裹药物小分子又具备较好水溶性，能够有效提高药物溶解度，提高药物稳定性能并掩盖一些天然提取物的不良气味，广泛应用于食品、保健以及医疗等领域。
环糊精溶液萃取法是一种从天然产物中提取活性成分的有效方法。环糊精与客体分子易形成包合物，可高效选择性包裹天然产物中的活性物质，并改善该客体分子的物理性质，如：溶解度、挥发性、亲水性质等。近年来，环糊精包合技术广泛应用于食品、保健以及医疗等领域。
超声辅助提取与其他非传统的牵引技术和超声波设备的简单扩展相比，其成本更低，目前已成为实验室和工业规模的成熟提取技术。其效率基于液相中微泡的生长和坍塌，同时形成极端温度和压力条件(5000℃和2000atm)导致分析样品的有效碎裂和解聚。因此，该技术可以高效提取目标化合物并缩短提取时间。
将超声辅助环糊精提取枇杷叶中活性物质，采用超临界气溶胶萃取技术干燥溶液，并结晶得到提取物纳米微胶囊，所述工艺的操作条件温和是一种绿色环保可循环的纳米微胶囊制备技术，可有效将有机溶剂排走，并制备得到粒度均一的枇杷叶提取物纳米微胶囊，所述纳米胶囊稳定性良好，能够掩盖提取物不良气味，具有良好的抗氧化活性活性。
本发明相比现有技术具有以下优点：本发明提供的制备方法简单易行，绿色环保。全程操作条件温和，提取阶段通入不活泼气体保护活性物质不被氧化，制备提取物纳米微胶囊阶段CO2气体性质稳定，避免提取物中抗氧化类物质被氧化，利用环糊精溶液作为提取剂可有效提取枇杷叶中活性成分且枇杷叶提取物纳米胶囊其中多酚类物质含量较高，得到产物有机溶剂残留极低。本发明为植物多酚提取及其制剂生产提供了参考依据，可有用于推广其他天然产物活性成分提取及制剂制备，枇杷叶提取物纳米胶囊中多酚含量较高，具有良好的抗氧化活性，可应用于食品、保健品、化妆品等行业。
附图说明
图1是枇杷叶提取物环糊精纳米微胶囊SEM电镜图；
图2是枇杷叶提取物纳米微胶囊粒度分布图。
具体实施方式
下面对本发明的实施例作详细说明，本实施例在以本发明技术方案为前提下进行实施，给出了详细的实施方式和具体的操作过程，但本发明的保护范围不限于下述的实施例。
实施例1
以α-CD溶液作为提取剂
本实施例以α-CD乙醇溶液为提取剂，采用超声辅助提取法提取枇杷叶中活性物质。取一定量α-CD溶于乙醇溶液得到环糊精浓度10mg/mL，将粉碎至100目的枇杷叶与环糊精溶液混合置于密闭容器并通入氮气保护，固液比1:15，超声条件设置为：超声温度55℃，超声功率240W，超声时间60min。提取液过滤，所得滤液通过毛细喷嘴喷入超临界结晶釜，工艺参数设置如下：压力10MPa，温度40℃，毛细喷嘴内径75μm，通过恒流泵以0.5mL/min将溶液泵入结晶釜，待溶液泵入结束，继续保持压力温度等参数，以1L/min继续通入CO2并从出口处排除带有有机溶剂的CO2 3h以上。测定多酚提取率、提取物清除DPPH自由基能力以及提取物环糊精纳米胶囊粒径。
实施例2
以β-CD溶液作作为提取剂
本实施例以β-CD乙醇溶液为提取剂，采用超声辅助提取法提取枇杷叶中活性物质。取一定量β-CD溶于乙醇溶液得到环糊精浓度10mg/mL，将粉碎至100目的枇杷叶与环糊精溶液混合置于密闭容器并通入氮气保护，固液比1:15，超声条件设置为：超声温度55℃，超声功率240W，超声时间60min。提取液过滤，所得滤液通过毛细喷嘴喷入超临界结晶釜，工艺参数设置如下：压力10MPa，温度40℃，毛细喷嘴内径75μm，通过恒流泵以0.5mL/min将溶液泵入结晶釜，待溶液泵入结束，继续保持压力温度等参数，以1L/min继续通入CO2并从出口处排除带有有机溶剂的CO2 3h以上。测定多酚提取率、提取物清除DPPH自由基能力以及提取物环糊精纳米胶囊粒径。
实施例3
以γ-CD溶液作作为提取剂
本实施例以γ-CD乙醇溶液为提取剂，采用超声辅助提取法提取枇杷叶中活性物质。取一定量γ-CD溶于乙醇溶液得到环糊精浓度10mg/mL，将粉碎至100目的枇杷叶与环糊精溶液混合置于密闭容器并通入氮气保护，固液比1:15，超声条件设置为：超声温度55℃，超声功率240W，超声时间60min。提取液过滤，所得滤液通过毛细喷嘴喷入超临界结晶釜，工艺参数设置如下：压力10MPa，温度40℃，毛细喷嘴内径75μm，通过恒流泵以0.5mL/min将溶液泵入结晶釜，待溶液泵入结束，继续保持压力温度等参数，以1L/min继续通入CO2并从出口处排除带有有机溶剂的CO2 3h以上。测定多酚提取率、提取物清除DPPH自由基能力以及提取物环糊精纳米胶囊粒径。
实施例4
羟丙基-β-CD溶液作作为提取剂
本实施例以羟丙基-β-CD乙醇溶液为提取剂，采用超声辅助提取法提取枇杷叶中活性物质。取一定量羟丙基-β-CD溶于乙醇溶液得到环糊精浓度10mg/mL，将粉碎至100目的枇杷叶与环糊精溶液混合置于密闭容器并通入氮气保护，固液比1:15，超声条件设置为：超声温度55℃，超声功率240W，超声时间60min。提取液过滤，所得滤液通过毛细喷嘴喷入超临界结晶釜，工艺参数设置如下：压力10MPa，温度40℃，毛细喷嘴内径75μm，通过恒流泵以0.5mL/min将溶液泵入结晶釜，待溶液泵入结束，继续保持压力温度等参数，以1L/min继续通入CO2并从出口处排除带有有机溶剂的CO2 3h以上。测定多酚提取率、提取物清除DPPH自由基能力以及提取物环糊精纳米胶囊粒径。
实施例5
羟乙基-β-CD溶液作作为提取剂
本实施例以羟乙基-β-CD乙醇溶液为提取剂，采用超声辅助提取法提取枇杷叶中活性物质。取一定量羟乙基-β-CD溶于乙醇溶液得到环糊精浓度10mg/mL，将粉碎至100目的枇杷叶与环糊精溶液混合置于密闭容器并通入氮气保护，固液比1:15，超声条件设置为：超声温度55℃，超声功率240W，超声时间60min。提取液过滤，所得滤液通过毛细喷嘴喷入超临界结晶釜，工艺参数设置如下：压力10MPa，温度40℃，毛细喷嘴内径75μm，通过恒流泵以0.5mL/min将溶液泵入结晶釜，待溶液泵入结束，继续保持压力温度等参数，以1L/min继续通入CO2并从出口处排除带有有机溶剂的CO2 3h以上。测定多酚提取率、提取物清除DPPH自由基能力以及提取物环糊精纳米胶囊粒径。实施例1-5所得提取率，DPPH自由基清除率以及提取物纳米微胶囊粒径见表1，以环糊精溶液作为多酚提取剂提取率较高(以没食子酸计)，抗氧化性能良好，产品粒径范围在300-700nm范围，产品粒度小且粒度分布均一。
表1不同环糊精溶液对提取率及抗氧化活性影响
Figure BDA0002315036800000051
实施例6
本实施例将羟丙基-β-CD分别溶于丙酮、二氯甲烷、二甲基亚砜中，考察在提取过程分别以丙酮、二氯甲烷、二甲基亚砜作为溶剂对产品性能的影响，将粉碎至100目的枇杷叶与羟丙基-β-CD溶液混合置于密闭容器并通入氮气保护，固液比1:15，超声条件设置为：超声温度55℃，超声功率240W，超声时间60min。提取液过滤，所得滤液通过毛细喷嘴喷入超临界结晶釜，工艺参数设置如下：压力10MPa,温度40℃，毛细喷嘴内径75μm，通过恒流泵以0.5mL/min将溶液泵入结晶釜，待溶液泵入结束，继续保持压力温度等参数，以1L/min继续通入CO2并从出口处排除带有有机溶剂的CO2 3h以上。测定多酚提取率、提取物清除DPPH自由基能力以及提取物环糊精纳米胶囊粒径，如表2所示以环糊精分别溶于丙酮、二氯甲烷以及二甲基亚砜不同有机溶剂作为多酚提取剂提取率范围在10～30mg/g(以没食子酸计)，抗氧化性能良好，产品粒径范围在500-900nm范围，产品粒度小且粒度分布均一。
表2不同有机溶剂对提取率及抗氧化活性影响
Figure BDA0002315036800000061
实施例7
本实施例以10mg/mL羟丙基-β-CD乙醇溶液作为提取剂提取枇杷叶活性成分，将粉碎后的枇杷叶与环糊精溶液混合置于密闭容器并通入氮气保护，固液比1:15，超声条件设置为：超声温度40℃，超声功率600W，超声时间30min。提取液过滤，所得滤液通过毛细喷嘴喷入超临界结晶釜，工艺参数设置如下：压力12MPa,温度35℃，毛细喷嘴内径75μm，通过恒流泵以0.5mL/min将溶液泵入结晶釜，待溶液泵入结束，继续保持压力温度等参数，以1L/min继续通入CO2并从出口处排除带有有机溶剂的CO2 3h以上。测得多酚提取率30.18mg/g、提取物清除DPPH自由基清除率82.18％以及提取物环糊精纳米胶囊粒径307.5nm。
实施例8
本实施例以10mg/mL羟丙基-β-CD乙醇溶液作为提取剂提取枇杷叶活性成分，将粉碎后的枇杷叶与环糊精溶液混合置于密闭容器并通入氮气保护，固液比1:10，超声条件设置为：超声温度35℃，超声功率800W，超声时间40min。提取液过滤，所得滤液通过毛细喷嘴喷入超临界结晶釜，工艺参数设置如下：压力16MPa,温度45℃，毛细喷嘴内径75μm，通过恒流泵以0.5mL/min将溶液泵入结晶釜，待溶液泵入结束，继续保持压力温度等参数，以1L/min继续通入CO2并从出口处排除带有有机溶剂的CO2 3h以上。测得多酚提取率34.24mg/g、提取物清除DPPH自由基清除率85.58％。图1为该工艺下制备得到产品SEM电镜图，图2为纳米微胶囊粒度分布图，观察纳米微胶囊呈无定型细小颗粒状，粒径为267.5nm，粒度较窄且粒度分布均一。
以上所述仅为本发明的较佳实施例而已，并不用以限制本发明，凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等，均应包含在本发明的保护范围之内。

Claims (1)

1.一种枇杷叶提取物纳米胶囊的制备方法，其特征在于，包括以下步骤：
(1)将羟丙基-β-CD溶解于乙醇中得到浓度10mg/mL的环糊精溶液作为提取剂；
(2)将粉碎后的枇杷叶粉末置于密闭容器中，添加环糊精溶液并通入氮气，固液比1:10，超声辅助提取，提取液过滤，收集滤液；其中，超声条件设置为：超声温度35℃，超声功率800W，超声时间40min；
(3)所得滤液通过毛细喷嘴喷入超临界结晶釜，工艺参数设置如下：压力16MPa，温度45℃，毛细喷嘴内径75μm，通过恒流泵以0.5mL/min将滤液泵入结晶釜，泵入结束，继续保持压力温度参数，以1L/min继续通入CO2并从结晶釜出口处排除带有有机溶剂的CO23h以上，卸压，打开结晶斧，收集所得固体，即为枇杷叶提取物纳米胶囊。
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