CN113367348B - 一种脂质体稳定的乳液及其制备方法 - Google Patents
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Abstract
本发明公开了一种脂质体稳定的乳液及其制备方法，涉及食品加工领域。本发明脂质体稳定的乳液的制备方法，包括以下步骤：制备脂质体薄膜，制备分散液，制备粗乳液，制备脂质体稳定的乳液。本发明通过对脂质体的组分进行合理配比，以及通过优化制备步骤及参数，提高了维生素E、维生素B2和β‑胡萝卜素的负载率，同时还提高了脂质体稳定的乳液的热力学稳定性，使其能够在超高温瞬时灭菌时保持稳定，使其中的维生素B2、维生素E和β‑胡萝卜素靶向运送，促进消化吸收，起到预防黄斑病变的作用，同时兼具有益的保健功能。
Description
技术领域
本发明涉及食品加工领域，特别是涉及一种脂质体稳定的乳液及其制备方法。
背景技术
黄斑区是视网膜的一个重要区域，位于眼后极部，主要与精细视觉及色觉等视功能有关。一旦黄斑区出现病变，常常出现视力下降、眼前黑影或视物变形。黄斑病变是一种老年人常见的影响黄斑的疾病，确切的病因尚不明。对65岁以上的人来说，黄斑病变是成为永远失明的第一原因。研究表明，维生素E可抑制眼睛晶状体内的过氧化脂反应，使末梢血管扩张，改善血液循环，预防近视的发生和发展，在视网膜健康和降低黄斑病变风险(AMD风险)方面起着一定作用；维生素B2可以促进机体发育和细胞再生，有增进视力的功效；β-胡萝卜素作为维生素A的前体，有补肝明目的作用，可治疗夜盲症。日常摄入足够的维生素E、维生素B2和β-胡萝卜素对预防黄斑病变起着很好的作用。
但是，维生素B2、维生素E和β-胡萝卜素是不稳定的，易受光、氧化和酸碱环境的影响，同时，维生素B2具有亲水性，而维生素E和β-胡萝卜素具有亲油性，导致其在食品应用方面受到限制。脂质体具有类似生物膜的闭合囊泡结构，可同时荷载亲水性和疏水性成分，并且，在消化过程中，脂质体在胃内可以保持完整，进入小肠后再进行生物活性物质的释放，提高了生物活性物质的生物利用度。脂质体稳定的乳液是构建了一种液体界面作为乳化剂的水包油(O/W)乳液，目前，现有技术中脂质体稳定的乳液存在负载率较低、热力学稳定性较差的问题，这限制了其在食品尤其是保健品，尤其是保健饮品中的应用。
如何使保健饮品中同时包含维生素B2、维生素E和β-胡萝卜素，起到预防黄斑病变的保健作用，是本领域一直未能解决的一个技术难题。
发明内容
本发明的目的是提供一种脂质体稳定的乳液及其制备方法，以解决上述现有技术存在的问题，使其同时包含维生素B2、维生素E和β-胡萝卜素，且保证其能够靶向运送，起到预防黄斑病变的作用。
为实现上述目的，本发明提供了如下方案：
本发明目的之一是提供一种脂质体稳定的乳液的制备方法，包括以下步骤：
步骤1，制备脂质体薄膜：将维生素E、磷脂、胆固醇和吐温80溶于乙醇中，旋蒸，得到脂质薄膜；
步骤2，制备分散液：将所述脂质体薄膜加入维生素B2的磷酸盐缓冲液中，超声波细胞破碎，得到分散液；
步骤3，制备粗乳液：将β-胡萝卜素和磷脂溶于油中得到油相，将分散液与去离子水混合得到水相，将所述油相和所述水相混合，高速分散，得到粗乳液；
步骤4，制备脂质体稳定的乳液：将所述粗乳液进行均质得到脂质体稳定的乳液。
进一步地，步骤1中所述磷脂、胆固醇和吐温80的质量比为4:1:2。
进一步地，步骤1中维生素E和磷脂的质量比为1:20。
进一步地，步骤2中所述磷酸盐缓冲液由Na2HPO4·12H2O和NaH2PO4·2H2O组成，pH为7.4，浓度为0.02mol/L；所述维生素B2与所述磷酸盐缓冲液的质量体积比为20mg:40mL。
进一步地，步骤2中所述细胞破碎每破碎3秒钟停1秒钟。
进一步地，步骤3中所述β-胡萝卜素、磷脂和油的质量体积比为20mg：200mg：1mL；所述分散液与所述去离子水的体积比为2-4:15；油相与水相的体积比为1:19。。
进一步地，步骤4中所述均质的条件为25℃、一级压力70MPa、二级压力700Mpa。
进一步地，所述维生素E、维生素B2和β-胡萝卜素的质量比为1:2:2。
本发明目的之二是提供一种利用上述制备方法制备得到的脂质体稳定的乳液。
本发明目的之三是提供上述乳液在制备保健饮品方面的应用。
本发明公开了以下技术效果：
本发明中的脂质体既起到传输系统的作用，又起到乳化剂的作用，提高了乳液的稳定性，通过对脂质体的组分进行合理配比，以及通过优化的制备步骤及参数，提高了维生素E、维生素B2和β-胡萝卜素的负载率，同时还提高了脂质体稳定的乳液的热力学稳定性，使其能够在超高温瞬时灭菌时保持稳定，使其中的维生素B2、维生素E和β-胡萝卜素靶向运送，促进消化吸收，起到预防黄斑病变的作用，同时兼具有益的保健功能。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案，下面将对实施例中所需要使用的附图作简单的介绍，显而易见地，下面描述中的附图仅仅是本发明的一些实施例，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他的附图。
图1为实施例1制得的功能性饮料的激光共聚焦显微观测图。
具体实施方式
现详细说明本发明的多种示例性实施方式，该详细说明不应认为是对本发明的限制，而应理解为是对本发明的某些方面、特性和实施方案的更详细的描述。
应理解本发明中所述的术语仅仅是为描述特别的实施方式，并非用于限制本发明。另外，对于本发明中的数值范围，应理解为还具体公开了该范围的上限和下限之间的每个中间值。在任何陈述值或陈述范围内的中间值以及任何其他陈述值或在所述范围内的中间值之间的每个较小的范围也包括在本发明内。这些较小范围的上限和下限可独立地包括或排除在范围内。
除非另有说明，否则本文使用的所有技术和科学术语具有本发明所述领域的常规技术人员通常理解的相同含义。虽然本发明仅描述了优选的方法和材料，但是在本发明的实施或测试中也可以使用与本文所述相似或等同的任何方法和材料。本说明书中提到的所有文献通过引用并入，用以公开和描述与所述文献相关的方法和/或材料。在与任何并入的文献冲突时，以本说明书的内容为准。
在不背离本发明的范围或精神的情况下，可对本发明说明书的具体实施方式做多种改进和变化，这对本领域技术人员而言是显而易见的。由本发明的说明书得到的其他实施方式对技术人员而言是显而易见的。本发明说明书和实施例仅是示例性的。
关于本文中所使用的“包含”、“包括”、“具有”、“含有”等等，均为开放性的用语，即意指包含但不限于。
本发明所用磷酸盐缓冲液由Na2HPO4·12H2O和NaH2PO4·2H2O组成，pH为7.4，浓度为0.02mol/L。
实施例1
步骤1，制备脂质体薄膜：将10mg维生素E、200mg磷脂、50mg胆固醇和100mg吐温80溶于20mL乙醇中，在40℃、0.085MPa条件下进行旋蒸，至乙醇全部蒸发，得到透明的脂质体薄膜；
步骤2，制备分散液：将20mg维生素B2溶于40mL磷酸盐缓冲液(0.02mol/L，pH＝7.4)中，加入步骤1制得的脂质体薄膜，在40℃，50％功率下细胞破碎15min(总功率450W，每破碎3s，停1s)至澄清透明，得到同时含有维生素E和维生素B2的分散液；
步骤3，制备粗乳液：将200mgβ-胡萝卜素和2g磷脂溶于10mL玉米油中，将40mL步骤2制得的分散液与150mL去离子水混合，利用高速分散机在10000rpm下分散5min得到粗乳液；
步骤4，制备脂质体稳定的乳液：将步骤3制得的粗乳液在25℃、一级压力70MPa、二级压力700Mpa的条件下，循环三次，进行高压均质，得到负载维生素E、维生素B2和β-胡萝卜素的脂质体稳定的乳液；
步骤5，制备功能性饮料：向步骤4制得的脂质体稳定的乳液中加入1g柠檬酸和10g木糖醇，并用水定容至1000mL，在25℃、一级压力70MPa、二级压力700Mpa的条件下，循环三次，进行高压均质，之后过滤、超高温瞬时灭菌(121℃，5s)、冷却、灌装得到功能性饮料。
对制得的功能性饮料进行激光共聚焦显微观测，结果如图1所示。由图1能够看出，本实施例制得的功能性饮料的乳液液滴为规则的球形，由于脂质体的亲水层包裹维生素B2，维生素B2自带荧光，使得乳液界面层排列规则球形，证明脂质体存在于乳液液滴表面并稳定乳液。
分别取步骤4制得的脂质体稳定的乳液(样品A)和步骤5制得的功能性饮料(样品B)各0.5mL分成两组，对维生素B2、维生素E和β-胡萝卜素负载率进行测定，具体测定步骤如下：
取0.5ml样品于10mL离心管中，依次加入2mL无水乙醇和3mL正己烷，在涡旋混合器上充分振荡，使乳液完全破乳。静置10min使得无水乙醇和正己烷分离，取正己烷相(上层)并收集。再加入3mL正己烷和2mL无水乙醇，重复三次上述操作，合并正己烷。分别在479nm、256nm用紫外可见分光光度计测定β-胡萝卜素、维生素E吸光度。取下层乙醇相在516nm激发波长，450-550nm发射波长下用荧光分光光度计测定维生素B2的荧光强度，并根据浓度标准曲线计算维生素B2、维生素E和β-胡萝卜素的浓度。负载率用实际载药量与实际加入量的比值表示，结果如表1所示。
表1
由表1能够看出，本发明制备的脂质体稳定的乳液对维生素B2、维生素E和β-胡萝卜素的包埋效果良好，维生素B2、维生素E和β-胡萝卜素的负载率均维持在高水平(>60％)；本发明所制得的功能性饮料在经过121℃，5s超高温灭菌的情况下，维生素B2、维生素E和β-胡萝卜素的负载率并未受到明显影响，证明本发明制备的脂质体稳定的乳液的热稳定性非常好。
实施例2
与实施例1不同之处在于，步骤3中分散液的加入量为20mL。
结果：对制得的功能性饮料进行激光共聚焦显微观测，结果与实施例1类似，这里不在重复附图。
实施例3
与实施例1不同之处在于，步骤3中分散液的加入量为25mL。
结果：对制得的功能性饮料进行激光共聚焦显微观测，结果与实施例1类似，这里不在重复附图。
实施例4
与实施例1不同之处在于，步骤3中分散液的加入量为30mL。
结果：对制得的功能性饮料进行激光共聚焦显微观测，结果与实施例1类似，这里不在重复附图。
实施例5
与实施例1不同之处在于，步骤3中分散液的加入量为35mL。
结果：对制得的功能性饮料进行激光共聚焦显微观测，结果与实施例1类似，这里不在重复附图。
以上所述的实施例仅是对本发明的优选方式进行描述，并非对本发明的范围进行限定，在不脱离本发明设计精神的前提下，本领域普通技术人员对本发明的技术方案做出的各种变形和改进，均应落入本发明权利要求书确定的保护范围内。
Claims (6)
1.一种脂质体稳定的乳液的制备方法，其特征在于，包括以下步骤：
步骤1，制备脂质体薄膜：将维生素E、磷脂、胆固醇和吐温80溶于乙醇中，旋蒸，得到脂质薄膜；
步骤2，制备分散液：将所述脂质体薄膜加入维生素B2的磷酸盐缓冲液中，超声波细胞破碎，得到分散液；
步骤3，制备粗乳液：将β-胡萝卜素和磷脂溶于油中得到油相，将分散液与去离子水混合得到水相，将所述油相和所述水相混合，高速分散，得到粗乳液；
步骤4，制备脂质体稳定的乳液：将所述粗乳液进行均质得到脂质体稳定的乳液；
步骤1中所述磷脂、胆固醇和吐温80的质量比为4:1:2；
步骤1中维生素E和磷脂的质量比为1:20；
所述维生素B2与所述磷酸盐缓冲液的质量体积比为20mg:40mL；
步骤3中所述β-胡萝卜素、磷脂和油的质量体积比为20mg：200mg：1mL；所述分散液与所述去离子水的体积比为2-4:15；油相与水相的体积比为1:19；
所述维生素E、维生素B2和β-胡萝卜素的质量比为1:2:2。
2.根据权利要求1所述的一种脂质体稳定的乳液的制备方法，其特征在于，步骤2中所述磷酸盐缓冲液由Na2HPO4·12H2O和NaH2PO4·2H2O组成，pH为7.4，浓度为0.02mol/L。
3.根据权利要求1所述的一种脂质体稳定的乳液的制备方法，其特征在于，步骤2中所述细胞破碎每破碎3秒钟停1秒钟。
4.根据权利要求1所述的一种脂质体稳定的乳液的制备方法，其特征在于，步骤4中所述均质的条件为25℃、一级压力70MPa、二级压力700Mpa。
5.根据权利要求1-4任一项所述的制备方法制备得到的脂质体稳定的乳液。
6.如权利要求5所述的乳液在制备保健饮品方面的应用。
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