CN109568293A - 一种虾青素生物微胶囊的制备方法 - Google Patents
一种虾青素生物微胶囊的制备方法 Download PDFInfo
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Abstract
本发明公开了一种虾青素生物微胶囊的制备方法,其通过生物壁材制备、芯材制备、包埋、洗涤、干燥等步骤制备得到虾青素微胶囊。通过该方法制备的虾青素微胶囊产品大小均一,安全性高,生物相容性好,易生物降解,成本低,并且有效提高了虾青素的稳定性,对生物材料的综合利用和虾青素的应用推广具有重要意义。
Description
技术领域
本发明属于生物活性物质制备技术领域,具体涉及一种虾青素生物微胶囊的制备方法。
背景技术
虾青素(Astaxanthin)又名虾黄质、龙虾壳色素,化学名称为3,3’-二羟基-β,β-胡萝卜素-4,4’-二酮,分子式为C40H52O4,相对分子质量为596.86,广泛存在于海洋动植物体内,因虾青素具有抗氧化、抗炎、着色、光保护、促进繁殖等多种生物学功能,近年来甚受国内外的食品、医药、化妆品、水产养殖等行业的关注。但由于虾青素化学稳定性差,在自然状态下极易发生氧化降解,使其应用开发受到很大限制。因此,如何保持虾青素及其产品的稳定性是一个有研究价值的问题,为此科研工作者做了很多工作。目前,有关该方面的报道主要有以下几个方面:(1)采用食品胶或环糊精为壁材,采用经喷雾干燥法制备虾青素微胶囊。然而,喷雾干燥对温度要求较高,极易导致虾青素发生热敏降解;(2)凝集法制备:通过生物大分子(主要是蛋白或多糖)为原料,利用两种带有相反电荷的高分子材料以离子间的作用相互交联,制成的复合型壁材的微胶囊。
发明专利CN 103875918 A公开了一种虾青素微胶囊制备工艺,纯胶囊和麦芽糊精1:1交合而成壁材加水混合成壁材溶液,再加入浓度为6%的乳化剂,制成壁材乳化剂溶液,再加入浓度为3%的虾青素油溶液,强烈搅拌成虾青素乳化剂,然后以30000r/min二次分散搅拌1min,以30000r/min三次分散搅拌1min,最后进行喷雾干燥制成虾青素胶囊。发明专利CN 104705654 A公开了一种虾青素微胶囊制备方法,具体过程为:首先将纯胶、麦芽糊精与蒸馏水搅拌均匀制得壁材溶液,然后将壁材溶液与复配乳化剂混合搅拌至完全乳化得壁材乳化剂溶液;再将雨生红球藻孢子破壁处理后采用常规的萃取工艺制得虾青素溶液,将虾青素溶液与维生素E混合得虾青素油溶液;再将壁材乳化剂溶液与虾青素油溶液混合并搅拌至有乳化剂形成为止,然后用超高细胞均质器均质,调节乳化剂温度后再用超高细胞均质器均质至其形成水色油微胶囊;最后用喷雾干燥机对水色油微胶囊进行喷雾干燥至壁材凝结,制得虾青素微胶囊。发明专利CN 104938885 A提供一种虾青素微胶囊制备的方法,其利用搅拌产生的机械外力和极性水产生的斥力将小分子虾青素推入β-环糊精锥形结构空腔内,形成β-环糊精微囊包合物半成品,然后利用黏度低,流动性好,成膜性强的阿拉伯胶对β-环糊精微囊包合物半成品进行包埋,最后经喷雾干燥制得虾青素微胶囊。
以上方法均具有一定的适用性,但同时存在着如成本高、安全性差、生物相容性差等问题。而本发明以生物材料作为壁材进行包埋,可以避免这些问题,有效扩宽了生物资源的综合利用途径和虾青素的应用范围。
发明内容
针对现有技术的不足,本发明通过提供一种虾青素生物微胶囊的制备方法,以解决传统工艺中易造成虾青素热降解,成本高,安全性差,生物相容性差等技术问题。
为实现上述目的,本发明的技术方案为:
一种虾青素生物微胶囊的制备方法,包括如下步骤:
(1)生物壁材的制备:取酵母菌干粉加入水制成酵母菌悬浮液,然后向悬浮液中加入酵母菌自溶促进剂使酵母菌自溶,自溶结束后进行过滤或离心,收集菌体,并用水洗涤1~3次,冷冻干燥,即得到生物壁材;
(2)芯材的制备:将虾青素溶解到有机溶液中,配成虾青素溶液,作为微胶囊的芯材;
(3)包埋:取步骤(2)制备的芯材,向其中加入步骤(1)制备的生物壁材,然后进行包埋;
(4)洗涤、干燥:包埋结束后进行离心或过滤,收集酵母菌体,然后用步骤(2)的有机溶液进行洗涤,除去酵母菌体表面残留的芯材,再经干燥制得虾青素生物微胶囊。
优选的,步骤(1)中,取酵母菌干粉按照质量比为1:3~4加入水制成酵母菌悬浮液;酵母菌自溶促进剂的添加量为酵母菌悬浊液质量的0.75~0.8%。
优选的,步骤(1)中,酵母菌自溶的温度条件为45℃~50℃,自溶时间为6~12h。
优选的,步骤(1)中,酵母菌为酿酒酵母和/或面包酵母;酵母菌自溶促进剂为为乙醇、乙酸、食盐、硫胺素、聚氧乙烯失水山梨醇脂肪酸酯中的一种或几种的混合物。
优选的,步骤(2)中,芯材中,虾青素的质量体积分数为4~5%。
优选的,步骤(2)中,所述的虾青素为游离状态的虾青素或酯化状态的虾青素;所述有机溶剂为乙醇、甘油、聚乙二醇、乙酸乙酯、油酸乙酯、甘油酯中的一种或几种的混合物。
优选的,步骤(2)中,所述有机溶剂为含水量在95%的乙醇溶液。
优选的,步骤(2)中,所述有机溶剂为含水量在90%的甘油溶液。
优选的,步骤(2)中,所述有机溶剂为含水量在1~20%的聚乙二醇溶液。
优选的,步骤(3)中,芯材与生物壁材的质量比为1:0.2-0.4;包埋条件为:温度40℃~45℃,压力为0~20M Pa,进行搅拌或震荡,频率为60~100r/min,时间为6~8h。
优选的,步骤(4)中,干燥方式为室温干燥,减压干燥、冷冻干燥中的一种;洗涤后的有机溶液返回步骤(2)进行循环利用。干燥方式优选冷冻干燥。
与现有技术相比,本发明的优点:
(1)本发明通过将虾青素芯材包埋于生物材料酵母菌中,制备得到具有良好生物相容性、安全性高的虾青素微胶囊,克服了传统虾青素微胶囊的缺点。
(2)本发明直接利用酵母细胞作为壁材,取代传统利用大分子材料作为壁材,产品安全性和生物相容性更好。
(3)本发明由于使用的的酵母细胞进行包裹,微胶囊产品的直径为5~10μm且大小均一,易生物降解,且酵母资源丰富,成本低。
利用本发明的方法制备的虾青素微胶囊且大小均匀。
(3)本发明的方法制备的虾青素微胶囊虾青素稳定性提高,保质期长。
(4)本发明的方法制备的虾青素微胶囊虾青素在水中具有良好的分散性。
(5)本发明的方法制备的虾青素微胶囊虾青素可广泛用于饲料、食品、化妆品和药品领域。
(6)本发明的制备方法工艺简单,条件温和,生产成本低,便于工业化推广,并且为酵母资源的综合利用提供了一条新途径,提高了资源利用率。
综上所述,通过本发明的方法制备的虾青素微胶囊产品大小均一,安全性高,生物相容性好,易生物降解,成本低,并且有效提高了虾青素的稳定性,对生物材料的综合利用和虾青素的应用推广具有重要意义。
附图说明
下面结合附图和实施例对本发明进一步说明。
图1是本发明的工艺流程图;
图2为由本发明的方法制备获得的虾青素生物微胶囊的显微观察结果;
图3为本发明制备的虾青素生物微胶囊的稳定性试验结果。
具体实施方式
本发明通过提供一种虾青素生物微胶囊的制备方法,充分发挥酵母菌作为天然生物壁材的优势,开发出一条制备过程简单、安全性高、成本低的虾青素微胶囊稳态化技术路线。本发明通过酵母自溶、芯材制备、洗涤、干燥,制备虾青素微胶囊的方法,可解决传统工艺中易造成虾青素热降解,成本高,安全性差,生物相容性差等问题,有效扩宽了生物资源的综合利用途径和虾青素的应用范围。
下面结合附图和具体实施方式对本发明作进一步详细的说明。
实施例1
一种虾青素生物微胶囊的制备方法,如图1所示的工艺流程,取1kg酿酒酵母干粉,加入3L水,制成酵母菌悬浮液,然后向悬浮液中加入30ml乙醇,在45℃温度条件下使酵母菌自溶,自溶时间为6h,自溶结束后进行离心,收集菌体,并用水洗涤2次,然后将菌体进行冷冻干燥,即得到生物壁材。
将虾青素按照质量比溶解到95%乙醇溶液中,配成质量体积分数为5%的虾青素溶液,作为微胶囊的芯材,向芯材中加入质量比为1:0.2的生物壁材,于40℃常压条件下震荡6h,震荡频率为80r/min,震荡结束后,离心,收集酵母菌体,然后用95%乙醇溶液进行洗涤,除去酵母表面残留的芯材,再经冷冻制得虾青素生物微胶囊,洗涤后的乙醇溶液返回进行循环利用。
(1)包埋率测定
虾青素微胶囊化效果可用包埋率来衡量,包埋率计算公式如下:
其中,M0为包埋之前芯材虾青素的重量(g),M1为未被包埋的芯材虾青素的重量(g)。通过测定,本实施例中虾青素生物微胶囊的包埋率为86.3%。
(2)虾青素生物微胶囊的形态观察
取本实施例制备的虾青素生物微胶囊,用水稀释100倍,制成虾青素生物微胶囊水分散系,然后取1滴与载玻片上,通过电子显微镜观察虾青素生物微胶囊的外观,并记录其直径大小,如附图2所示,观察结果显示本发明制备的虾青素生物微胶囊外观为圆球形(或为蛋形),大小约为5~10μm。
(3)虾青素生物微胶囊的稳定性检验
取相同重量的虾青素粉末和本实施例制备的虾青素生物微胶囊,于55℃光照培养箱中放置,贮藏过程中通过分光光度法测量虾青素的损失率,进而评价其稳定性。具体测定方法如下:
在贮藏过程中,不定期准确取2mg虾青素粉末和本实施例制备的虾青素生物微胶囊,用二甲基亚砜定容至50ml,在489nm下的测定吸光值,计算虾青素损失率,评价其稳定性。
虾青素损失率计算公式:
其中:A0为贮藏期开始时样品溶液在489nm下的吸光度值;Ai为贮藏第i天时样品溶液在489nm下的吸光度值。
稳定性检验结果如图3所示,通过图中结果表明,虾青素粉末在贮藏至第7天损失率为96%,而虾青素生物微胶囊在贮藏至第7天损失率为6%。由此可见,利用生物壁材制备虾青素微胶囊可显著提高虾青素的贮藏稳定性,为提高虾青素的商业应用价值具有重要意义。
实施例2
一种虾青素生物微胶囊的制备方法,取1kg面包酵母干粉,加入4L水,制成酵母菌悬浮液,然后向悬浮液中加入酵母菌40ml吐温80,在50℃温度条件下使酵母菌自溶,自溶时间为8h,自溶结束后进行离心,收集菌体,并用水洗涤1次,然后将菌体进行冷冻干燥,即得到生物壁材。
将虾青素按照质量比溶解到90%甘油溶液中,,配成质量体积分数为4%的虾青素溶液,作为微胶囊的芯材,向芯材中加入质量比为1:0.3的生物壁材,于45℃,20Mpa压力条件下搅拌6h,搅拌转速为60r/min,结束后,过滤,收集酵母菌体,然后用90%甘油溶液进行洗涤,除去酵母表面残留的芯材,再经冷冻制得虾青素生物微胶囊,洗涤后的甘油溶液返回进行循环利用。
通过测定,本实施例制备的虾青素生物微胶囊的包埋率为87.5%,其在显微镜下外观呈圆球形(或为蛋形),大小约为5~10μm。
实施例3
一种虾青素生物微胶囊的制备方法,取1kg酿酒酵母干粉,加入3L水,制成酵母菌悬浮液,然后向悬浮液中加入酵母菌30ml乙酸乙酯,在45℃温度条件下使酵母菌自溶,自溶时间为12h,自溶结束后进行过滤,收集菌体,并用水洗涤3次,然后将菌体进行冷冻干燥,即得到生物壁材。
将酯化形态的虾青素按照质量比溶解到乙酸乙酯溶液中,配成质量体积分数为5%的酯化虾青素溶液,作为微胶囊的芯材,向芯材中加入质量比为1:0.4的生物壁材,于40℃常压条件下震荡8h,震荡频率为100r/min,震荡结束后,过滤,收集酵母菌体,然后用乙酸乙酯进行洗涤,除去酵母表面残留的芯材,再经冷冻制得虾青素生物微胶囊,洗涤后的乙醇溶液返回进行循环利用。
通过测定,本实施例制备的虾青素生物微胶囊的包埋率为85.9%,其在显微镜下外观呈圆球形(或为蛋形),大小约为5~10μm。
虽然,上文中已经用一般性说明及具体实施例对本发明作了详尽的描述,但在本发明基础上,可以对之作一些修改或改进,这对本领域技术人员而言是显而易见的。因此,在不偏离本发明精神的基础上所做的这些修改或改进,均属于本发明要求保护的范围。
Claims (10)
1.一种虾青素生物微胶囊的制备方法,其特征在于,包括如下步骤:
(1)生物壁材的制备:取酵母菌干粉加入水制成酵母菌悬浮液,然后向悬浮液中加入酵母菌自溶促进剂使酵母菌自溶,自溶结束后进行过滤或离心,收集菌体,并用水洗涤1~3次,冷冻干燥,即得到生物壁材;
(2)芯材的制备:将虾青素溶解到有机溶液中,配成虾青素溶液,作为微胶囊的芯材;
(3)包埋:取步骤(2)制备的芯材,向其中加入步骤(1)制备的生物壁材,然后进行包埋;
(4)洗涤、干燥:包埋结束后进行离心或过滤,收集酵母菌体,然后用步骤(2)的有机溶液进行洗涤,除去酵母菌体表面残留的芯材,再经干燥制得虾青素生物微胶囊。
2.根据权利要求1所述的虾青素生物微胶囊的制备方法,其特征在于:步骤(1)中,取酵母菌干粉按照质量比为1:3~4加入水制成酵母菌悬浮液;酵母菌自溶促进剂的添加量为酵母菌悬浊液质量的0.75~0.8%。
3.根据权利要求2所述的虾青素生物微胶囊的制备方法,其特征在于:步骤(1)中,酵母菌自溶的温度条件为45℃~50℃,自溶时间为6~12h。
4.根据权利要求2或3所述的虾青素生物微胶囊的制备方法,其特征在于:步骤(1)中,酵母菌为酿酒酵母和/或面包酵母;酵母菌自溶促进剂为为乙醇、乙酸、食盐、硫胺素、聚氧乙烯失水山梨醇脂肪酸酯中的一种或几种的混合物。
5.根据权利要求2或3所述的虾青素生物微胶囊的制备方法,其特征在于:步骤(2)中,芯材中,虾青素的质量体积分数为4~5%。
6.根据权利要求5所述的虾青素生物微胶囊的制备方法,其特征在于:步骤(2)中,所述的虾青素为游离状态的虾青素或酯化状态的虾青素;所述有机溶剂为乙醇、甘油、聚乙二醇、乙酸乙酯、油酸乙酯、甘油酯中的一种或几种的混合物。
7.根据权利要求6所述的虾青素生物微胶囊的制备方法,其特征在于:步骤(2)中,所述有机溶剂为含水量在95%的乙醇溶液。
8.根据权利要求6所述的虾青素生物微胶囊的制备方法,其特征在于:步骤(2)中,所述有机溶剂为含水量在90%的甘油溶液。
9.根据权利要求5所述的虾青素生物微胶囊的制备方法,其特征在于:步骤(3)中,芯材与生物壁材的质量比为1:0.2-0.4;包埋条件为:温度40℃~45℃,压力为0~20M Pa,进行搅拌或震荡,频率为60~100r/min,时间为6~8h。
10.根据权利要求9所述的虾青素生物微胶囊的制备方法,其特征在于:步骤(4)中,干燥方式为室温干燥,减压干燥、冷冻干燥中的一种;洗涤后的有机溶液返回步骤(2)进行循环利用。
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