CN111387339A - 一种具有仿母乳脂肪球结构的大尺寸脂质体及其制备方法 - Google Patents
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Abstract
本发明公开了一种具有仿母乳脂肪球结构的大尺寸脂质体及其制备方法。所述脂质体由磷脂、缓冲液、水溶性营养素和/或油溶性营养素组成，采用平板堆彻自组装结合超声驱动技术，制备过程主要包括以下步骤：脂质溶解、平板涂层堆彻、自组装、超声驱动。本发明所得大尺寸脂质体粒径可控，尺寸、膜结构接近母乳脂肪球粒径和外层双分子层膜，载药空间较大，易被人体接受。
Description
技术领域
本发明属于食品技术领域，特别是涉及一种具有仿母乳脂肪球结构的大尺寸脂质体及其制备方法。
背景技术
脂质体是一种人工双分子层膜。当两性分子如磷脂和鞘脂分散于水相时，分子的疏水尾部倾向于聚集在一起，避开水相，而亲水头部暴露在水相，形成具有双分子层结构的封闭囊泡，称为脂质体。脂质体最初用于基因治疗和药物靶向运输，可与细胞膜融合，将药物运送至细胞内部。近年来，脂质体已逐渐应用于食品领域，包埋功能蛋白/多肽/酶、多酚/黄酮类、功能性脂肪酸、维生素和矿物质等，应用于乳制品、饮料、肉制品和巧克力等，可改善不良风味、延长食品货架期、提高营养素吸收率目的。目前，脂质体的制备方法主要有薄膜分散法、逆向蒸发法、高压均质法及其结合法，但这些传统方法制得的脂质体粒径分布不均匀、包埋率有限、存在有机溶剂残留等问题，影响了脂质体的广泛应用。
母乳中的脂肪球(MFG)平均粒径约为4μm，MFG以结构不同的甘油三酯为核心，最外层主要以双层生物分子膜结构的乳脂肪球膜磷脂(MFGM)组成，起到乳化和提供生物活性物质的作用，可防止脂滴聚结和内源性脂肪酶的水解。婴儿配方奶粉的生产需经过原料乳脱脂、添加乳脂配料、均质化、高温杀菌和喷雾干燥等多道工序，MFG的平均直径减小至母乳MFG的1/10(0.3-1.0μm)；MFGM亦大量损失，取而代之的是以蛋白质为主的乳化剂，所承运的营养因子锐减。因此，婴儿对母乳和配方奶粉的脂质消化和吸收差异显著。
制备具有仿母乳脂肪球粒径和表面结构的脂质体，可以提高人体对脂质体的接收程度；同时，相较于传统脂质体，较大的粒径意味着有更大的运载空间，提高了脂质体载药量。为此，开发具有仿母乳脂肪球结构的大尺寸脂质体及其制备方法意义重大。
发明内容
本发明的目的在于提供一种具有仿母乳脂肪球结构的大尺寸脂质体及其制备方法。本发明制备了一种具有仿母乳脂肪球结构的大尺寸脂质体，该脂质体尺寸、膜结构接近母乳脂肪球粒径和外层双分子层薄膜，易于人体接收，可提高所包埋的芯材的稳定性和提高生物利用度，适宜添加入婴儿配方奶粉及其它辅食，具有较好的实际使用价值。
为了达到上述目的，本发明采取以下技术方案：
一种具有仿母乳脂肪球结构的大尺寸脂质体的制备方法，采用平板堆彻自组装结合超声驱动技术，包括如下步骤：
(1)脂质溶解：向磷脂中加入无水乙醇，搅拌均匀至完全溶解；
(2)平板涂层堆砌：将步骤(1)中所述脂质溶液均匀涂层于平板，待乙醇挥发形成磷脂薄膜均匀堆彻于平板，再次涂层，直至平板上形成多层脂质堆彻的均匀薄膜；
(3)自组装：向步骤(2)中所述脂质薄膜加入缓冲液，随后放入空气浴摇床中震摇，脂质薄膜吸胀，自组装形成粒径较大的粗脂质体悬液；
(4)超声驱动：将步骤(3)得到的粗脂质体悬液进行超声，通过控制时间和功率，驱使脂质体二次组装，形成具有仿母乳脂肪球结构的大尺寸(3-4μm)脂质体；
优选地，步骤(1)中根据需要包埋的营养素的性质，在无水乙醇中加入脂溶性营养素。
优选地，步骤(1)中，磷脂为磷脂酰胆碱、磷脂酰乙醇胺、磷脂酰丝氨酸、磷脂酰肌醇、磷脂酰甘油、甘油磷脂酸等常见磷脂中的一种或多种。
优选地，步骤(3)中，缓冲液为HEPES缓冲液、PBS缓冲液、柠檬酸缓冲液中的一种。
优选地，步骤(3)中，根据需要包埋的营养素的性质，在缓冲液中加入水溶性营养素。
优选地，步骤(3)脂质体悬液中磷脂浓度为5-50mg/mL。
优选地，步骤(3)震荡温度为45-60℃。
优选地，步骤(4)超声功率为100-300w，超声时间为10-50s。
本发明还提供了一种由上述制备方法获得的具有仿母乳脂肪球结构的大尺寸脂质体。
本发明采用以平板为界面的平板堆彻自组装结合超声驱动技术制备了一种具有仿母乳脂肪球结构的大尺寸脂质体。将磷脂和油溶性物质均匀分散在平板上涂层堆砌形成薄膜，再以溶解有水溶性芯材的缓冲液洗膜，经过自组装形成粗脂质体，最后经过一定条件的超声驱动，形成具有母乳脂肪球结构的大尺寸脂质体。
本发明具有以下技术特点：
1)本发明建立了一种新型的脂质体制备方法——平板堆彻自组装结合超声驱动技术，操作简便，设备简单，绿色无污染；
2)本发明获得的大尺寸脂质体与母乳脂肪球尺寸接近，且粒径分布均匀，膜结构与母乳脂肪球外双分子层相似，包埋营养素比例高，可应用于婴儿强化食品。
附图说明
图1是本发明的工艺流程图；
图2是本发明一种具有仿母乳脂肪球结构的大尺寸脂质体一较佳实施例(实施例1)的微观形貌图
图3是具体实施方式中实施例1-2制备得到的具有仿母乳脂肪球结构的大尺寸脂质体的粒径D(3,2)示意图；
图4是具体实施方式中实施例1-2制备得到的具有仿母乳脂肪球结构的大尺寸脂质体的电位示意图。
具体实施方式
以下具体实施例是对本发明提供的方法与技术方案的进一步说明，但不应理解成对本发明的限制。
本发明公开了一种具有仿母乳脂肪球结构的大尺寸脂质体，本领域技术人员可以借鉴本文内容，适当改进工艺参数实现。特别需要指出的是，所有类似的替换和改动对本领域技术人员来说是显而易见的，它们都被视为包括在本发明。本发明所述具有仿母乳脂肪球结构的大尺寸脂质体已经通过较佳实施例进行了描述，相关人员明显能在不脱离本发明内容、精神和范围内对本文所述的方法和应用进行改动或适当变更与组合，来实现和应用本发明技术。
一种具有仿母乳脂肪球结构的大尺寸脂质体，所述大尺寸脂质体主要由磷脂、缓冲液组成；其中，根据实际需要，可以在制备过程中加入油溶性和/或水溶性物质作为芯材。
如图1所示，是本发明一种具有仿母乳脂肪球结构的大尺寸脂质体的制备方法流程图，主要包括如下步骤：
(1)脂质溶解：向磷脂中加入无水乙醇，搅拌均匀至完全溶解；
(2)平板涂层堆砌：将步骤(1)中所述脂质溶液均匀涂层于平板，待乙醇挥发形成磷脂薄膜均匀堆彻于平板，再次涂层，直至平板上形成多层脂质堆彻的均匀薄膜；
(3)自组装：向步骤(2)中所述脂质薄膜加入缓冲液，随后放入空气浴摇床中震摇，脂质薄膜吸胀，自组装形成粒径较大的粗脂质体悬液；
(4)超声驱动：将步骤(3)得到的粗脂质体悬液进行超声，通过控制时间和功率，驱使脂质体二次组装，形成具有仿母乳脂肪球结构的大尺寸(3-4μm)脂质体；
下面结合实施例，进一步阐述本发明。
实施例1：
(1)脂质溶解：称取80mg磷脂酰胆碱，加入2mL无水乙醇，搅拌均匀至完全溶解；
(2)平板涂层堆砌：将步骤(1)中所述脂质溶液均匀涂层于平板，待乙醇挥发形成磷脂薄膜均匀堆彻于平板，再次涂层，直至平板上形成多层脂质堆彻的均匀薄膜；
(3)自组装：向步骤(2)中所述脂质薄膜加入10mL 0.05M HEPES缓冲液，随后放入55℃空气浴摇床中震摇16h，自组装形成粒径较大的粗脂质体悬液，磷脂浓度为8mg/mL；
(4)超声驱动：将步骤(3)得到的粗脂质体悬液以210w功率进行超声20s，驱使脂质体二次组装，形成具有仿母乳脂肪球结构的大尺寸(3-4μm)脂质体。
将得到的具有仿母乳脂肪球结构的大尺寸脂质体进行储存、检验。得到的具有仿母乳脂肪球粒径的大尺寸脂质体的微观形貌图，如图2所示。通过平均粒径和zeta电位检测，如图3、4所示，所得具有仿母乳脂肪球结构的大尺寸脂质体粒度D(3,2)为3.51±0.12μm，zeta电位为-4.55±0.07mV。
实施例2：
(1)脂质溶解：称取100mg磷脂酰胆碱，加入2mL无水乙醇，搅拌均匀至完全溶解；
(2)平板涂层堆砌：将步骤(1)中所述脂质溶液均匀涂层于平板，待乙醇挥发形成磷脂薄膜均匀堆彻于平板，再次涂层，直至平板上形成多层脂质堆彻的均匀薄膜；
(3)自组装：向步骤(2)中所述脂质薄膜加入10mL含有2mg/mL乳铁蛋白的0.05MPBS缓冲液，随后放入55℃空气浴摇床中震摇12h，脂质薄膜吸胀，自组装形成粒径较大的粗脂质体悬液,磷脂浓度为10mg/mL；
(4)超声驱动：将步骤(3)得到的粗脂质体悬液以210w功率进行超声25s，驱使脂质体二次组装，形成具有仿母乳脂肪球结构的大尺寸(3-4μm)脂质体。
将得到的具有仿母乳脂肪球结构的大尺寸脂质体进行储存、检验。通过粒度和zeta电位检测，如图3、4所示，所得具有仿母乳脂肪球结构的大尺寸脂质体粒度D(3,2)为3.31±0.10μm，zeta电位为-6.55±0.27mV。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。
Claims (9)
1.一种具有仿母乳脂肪球结构的大尺寸脂质体的制备方法，采用平板堆彻自组装结合超声驱动技术，包括如下步骤：
(1)脂质溶解：向磷脂中加入无水乙醇，搅拌均匀至完全溶解获得脂质溶液；
(2)平板涂层堆砌：将步骤(1)中所述脂质溶液均匀涂层于平板，待乙醇挥发形成磷脂薄膜均匀堆彻于平板，再次涂层，直至平板上形成多层脂质堆彻的均匀薄膜；
(3)自组装：向步骤(2)中脂质薄膜加入缓冲液，随后放入空气浴摇床中震摇，脂质薄膜吸胀，自组装形成粒径较大的粗脂质体悬液；
(4)超声驱动：将步骤(3)得到的粗脂质体悬液进行超声，驱使脂质体二次组装，形成具有仿母乳脂肪球结构的大尺寸(3-4μm)脂质体。
2.根据权利要求1所述的制备方法，其特征在于，步骤(1)中，根据需要包埋的营养素的性质，在无水乙醇中加入脂溶性营养素。
3.根据权利要求1所述的制备方法，其特征在于，步骤(1)中，磷脂为磷脂酰胆碱、磷脂酰乙醇胺、磷脂酰丝氨酸、磷脂酰肌醇、磷脂酰甘油、甘油磷脂酸等常见磷脂中的一种或多种。
4.根据权利要求1所述的制备方法，其特征在于，步骤(3)中，缓冲液为HEPES缓冲液、PBS缓冲液、柠檬酸缓冲液中的一种。
5.根据权利要求1所述的制备方法，其特征在于，步骤(3)中，根据需要包埋的营养素的性质，在缓冲液中加入水溶性营养素。
6.根据权利要求1所述的制备方法，其特征在于，步骤(3)脂质体悬液中磷脂浓度为5-50mg/mL。
7.根据权利要求1所述的制备方法，其特征在于，步骤(3)震荡温度为45-60℃。
8.根据权利要求1所述的制备方法，其特征在于，步骤(4)超声功率为100-300w，超声时间为10-50s。
9.由权利要求1-7任一所述制备方法获得的具有仿母乳脂肪球结构的大尺寸脂质体。
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1991010422A1 (en) * | 1990-01-12 | 1991-07-25 | The Liposome Company, Inc. | Interdigitation-fusion liposomes and gels |
| JPH0624999A (ja) * | 1992-07-07 | 1994-02-01 | Nippon Oil & Fats Co Ltd | リポソーム製剤 |
| WO2005051091A1 (en) * | 2003-10-22 | 2005-06-09 | Enzymotec Ltd. | Mimetic lipids and dietary supplements comprising the same |
| US20070065541A1 (en) * | 1997-11-05 | 2007-03-22 | Biozone Laboratories, Inc. | Enhanced infant formula containing liposome encapsulated nutrients and agents |
| US20170042809A1 (en) * | 2015-08-11 | 2017-02-16 | Peregrine Ophthalmic PTE LTD. | Stable liposomal formulations of carbonic anhydrase inhibitors for ocular drug delivery |
| CN107496359A (zh) * | 2017-10-09 | 2017-12-22 | 江西科技师范大学 | PluronicF127姜黄素脂质体的制备方法 |
| CN109771373A (zh) * | 2019-01-23 | 2019-05-21 | 浙江工商大学 | 脂质体稳定的水包油Pickering乳液及其制备方法 |
| CN110742130A (zh) * | 2019-11-14 | 2020-02-04 | 东北农业大学 | 一种添加模拟母乳脂肪结构的脂质体的婴幼儿配方羊奶粉及其制备方法 |
-
2020
- 2020-02-20 CN CN202010104418.2A patent/CN111387339B/zh active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1991010422A1 (en) * | 1990-01-12 | 1991-07-25 | The Liposome Company, Inc. | Interdigitation-fusion liposomes and gels |
| JPH0624999A (ja) * | 1992-07-07 | 1994-02-01 | Nippon Oil & Fats Co Ltd | リポソーム製剤 |
| US20070065541A1 (en) * | 1997-11-05 | 2007-03-22 | Biozone Laboratories, Inc. | Enhanced infant formula containing liposome encapsulated nutrients and agents |
| WO2005051091A1 (en) * | 2003-10-22 | 2005-06-09 | Enzymotec Ltd. | Mimetic lipids and dietary supplements comprising the same |
| US20170042809A1 (en) * | 2015-08-11 | 2017-02-16 | Peregrine Ophthalmic PTE LTD. | Stable liposomal formulations of carbonic anhydrase inhibitors for ocular drug delivery |
| CN107496359A (zh) * | 2017-10-09 | 2017-12-22 | 江西科技师范大学 | PluronicF127姜黄素脂质体的制备方法 |
| CN109771373A (zh) * | 2019-01-23 | 2019-05-21 | 浙江工商大学 | 脂质体稳定的水包油Pickering乳液及其制备方法 |
| CN110742130A (zh) * | 2019-11-14 | 2020-02-04 | 东北农业大学 | 一种添加模拟母乳脂肪结构的脂质体的婴幼儿配方羊奶粉及其制备方法 |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116106231A (zh) * | 2023-04-13 | 2023-05-12 | 北京三元食品股份有限公司 | 一种多维度评价样品与母乳相似度的方法 |
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